Notice of Pre-AIA or AIA Status
The present application, filed on or after March 16, 2013, is being examined under the first inventor to file provisions of the AIA .
DETAILED ACTION
Claims 57-59, 61, 64-65, and 67-78 are pending in the instant application.
Claims 60, 62-63, and 66 are canceled.
Claims 77-78 are new.
Claim Objections and Rejections Withdrawn
The rejections to claims 60, 62-63, and 66 are moot in view of claim cancelation.
The objection to claim 75 is withdrawn in view of claim amendment.
The rejections to claims 57-59, 61, 64-65, and 67-76 under 35 USC §112(b) are
withdrawn in view of claim amendment.
The rejections to claims 57-59, 61, 64-65, and 67-76 under 35 USC §103 are withdrawn in view of claim amendment.
The rejections to claims 57-59, 61, 64-65, and 67-76 under Nonstatutory Double Patenting are withdrawn in view of claim amendment.
Claim Rejections – 35 USC § 103
In the event the determination of the status of the application as subject to AIA 35 U.S.C. 102 and 103 (or as subject to pre-AIA 35 U.S.C. 102 and 103) is incorrect, any correction of the statutory basis (i.e., changing from AIA to pre-AIA ) for the rejection will not be considered a new ground of rejection if the prior art relied upon, and the rationale supporting the rejection, would be the same under either status.
The following is a quotation of 35 U.S.C. 103 which forms the basis for all obviousness rejections set forth in this Office action:
A patent for a claimed invention may not be obtained, notwithstanding that the claimed invention is not identically disclosed as set forth in section 102, if the differences between the claimed invention and the prior art are such that the claimed invention as a whole would have been obvious before the effective filing date of the claimed invention to a person having ordinary skill in the art to which the claimed invention pertains. Patentability shall not be negated by the manner in which the invention was made.
This application currently names joint inventors. In considering patentability of the claims the examiner presumes that the subject matter of the various claims was commonly owned as of the effective filing date of the claimed invention(s) absent any evidence to the contrary. Applicant is advised of the obligation under 37 CFR 1.56 to point out the inventor and effective filing dates of each claim that was not commonly owned as of the effective filing date of the later invention in order for the examiner to consider the applicability of 35 U.S.C. 102(b)(2)(C) for any potential 35 U.S.C. 102(a)(2) prior art against the later invention.
The factual inquiries for establishing a background for determining obviousness under 35 U.S.C. 103 are summarized as follows:
1. Determining the scope and contents of the prior art.
2. Ascertaining the differences between the prior art and the claims at issue.
3. Resolving the level of ordinary skill in the pertinent art.
4. Considering objective evidence present in the application indicating obviousness or nonobviousness.
Claims 57-59, 61, 64-65, 67-68, and 70-78 are rejected under 35 U.S.C. 103 as being unpatentable over US 2017/0281795 (Geall AJ et al. reference of record), Heukers R et al. (Nanomedicine 2014 10(7):1441-51 reference of record), Niewoehner J et al. (Neuron 2014 81(1):49-60. doi: 10.1016/j.neuron.2013.10.061.), Weng A et al. (Journal of Controlled Release 2015 206 75-90 reference of record), Firestone RA e al. (Journal of Controlled Release 1996 39 (2-3) 251-259 reference of record), and US 2004/0242502 (Marciani D et al. reference of record).
Regarding instant claims 57-59, 64-65, 67-68, 70-75, and 77-78, Geall taught an effective method of cancer treatment by silencing a protein in a cancer cell comprising administering a single pharmaceutical combination comprising:
a first conjugate comprising an EGFR targeted antibody covalently linked to an effector molecule wherein the effector molecule is an siRNA oligonucleotide; and
a second conjugate comprising an EGFR targeted antibody covalently linked to melittin (page 148, [0689]),
wherein melittin is an endosomolytic moiety that causes endosomal escape agent (page 148, [0689]), wherein the expression of KRAS in the tumor was effectively silenced from 24 to 168 hours and wherein KRAS was decreased more in tumors that were administered the combination compared to an EGFR targeted antibody covalently linked to the siRNA alone (Fig. 33). Regarding claims 57-59, 64-65, 67-68, 70-75, and 77-78, Geall taught the antibody as a single-domain antibody (sdAb) (page 1, [0019]). Regarding claims 57-59, 64-65, 67-68, 70-75, and 77-78, Geall taught the endosomolytic moiety as an endosomolytic small molecule and an endosomolytic polymer (page 73, [0389]). Geall taught targeting CD71 with an antibody siRNA conjugate (Fig. 54), wherein the CD71 targeting antibody siRNA conjugate effectively silenced HPRT expression (Fig. 86).
Geall did not teach a pharmaceutical composition of a first conjugate comprising a sdAb-siRNA effector and a saponin that was a monodesmosidic triterpene glycoside or a bidesmosidic triterpene glycoside with a hydrazone bond conjugated to the endosomal escape moiety at a C23 of an aglycone core, but this is obvious in view of Heukers, Niewoehner, Weng, Firestone, and Marciani.
Heukers taught EGFR-targeted nanobody® VHH single domain conjugates are specific and potently kill cancer cells (abstract and Fig. 2). Heukers taught a biparatopic EGFR-targeted single domain antibody 7D12-9G8 that is known to be internalized (page 1442, left column, first paragraph). Heukers taught 7D12-9G8-PS induces the lysosomal trafficking (page, 1448, right column first paragraph). Heukers taught EGFR-targeted single domain antibodies showed a faster accumulation at the tumor, a more homogeneous distribution within the tumor, and a more rapid clearance of unbound molecules, compared to an anti-EGFR monoclonal antibody (page 1442, left column, first paragraph).
Niewoehner taught a bivalent transferrin receptor (TfR or CD71) antibody leads to effective lysosomal targeting, wherein the bivalent TfR antibody is the same (abstract and Fig. 2E).
Weng taught the glycoside saponin SO1861 greatly improves the efficacy of lipid based as well as non-lipid based targeted nanoplexes consisting of a targeted K16 peptide with a nucleic acid binding domain and plasmid-DNA, minicircle-DNA or small interfering RNA (siRNA) (abstract). Weng taught SO1861 augments the escape of the genetic cargo out of the intracellular compartments into the cytosol (abstract). Weng taught SO1861 induces the release of the genetic cargo out of endosomes and lysosomes (abstract), wherein the lumen of the endosomes and lysosomes are acidified (page 75, right column, last paragraph). Weng taught that SO1861 integrated into a receptor targeted nanoplex improved DNA transfection and provides improved effector properties (page 85, right column, last paragraph and Fig. 10). Weng taught that SO1861 in combination with siRNA increased siRNA silencing in cancer cells (page 85, left to right column, bridging paragraph and Fig. 9).
Weng taught an effective in vitro method for transferring an oligonucleotide effector molecule from outside a Neuro-2A cell to inside of said cell comprising:
providing a pharmaceutical composition comprising a conjugate comprising SO1861 sensitized receptor-targeted nanoplexes and an oligonucleotide effector of EGFP;
providing a Neuro-2A cells wherein the surface of the cell is targeted by the receptor-targeted nanoplexes;
contacting the cell of step b) with the pharmaceutical composition of step a),
therewith establishing the transfer of the oligonucleotide effector from outside the cell into the cell, wherein the integration of SO1861 into the nanoplex caused a tremendous shift of the median fluorescence value, and wherein the high transfection efficiency
of the SO1861 sensitized LPDS-nanoplexes is corroborated by the fluorescence
image (page 85, right column, last paragraph, Fig. 10 legend, and Fig. 10).
Weng taught the RISC (RNA-Induced-Silencing Complex) is located in the cytosol (page 75-76 bridging paragraph). Weng taught to exert a silencing effect, siRNA has to escape from the endosomes/lysosomes into the cytosol because in lysosomes siRNA/DNA are degraded by nucleases (page 75-76 bridging paragraph). Weng taught: 1) If the endo-/lysosomal escape of the genetic cargo does not occur the siRNA/DNA is degraded within the lysosome thus leading to a therapeutic failure; 2) limited release of siRNA/DNA into the cytosol is one of the major obstacles for efficient siRNA/DNA delivery; and 3) development of innovative strategies to augment the endo-/lysosomal escape of siRNA/DNA and the delivery of a minimal amount of siRNA/DNA are desired for an efficient therapeutic response (page 75-76 bridging paragraph).
Firestone taught BR96-Dox is an immunoconjugate in which doxorubicin is linked via an acid-labile hydrazone to the chimeric MAb BR96 (abstract). Firestone taught BR96 bound to tumor cells, then internalized via endocytosis into lysosomes, wherein the acidic milieu of the lysozymes hydrolyzed the hydrazone linkage, releasing free Dox (abstract). Firestone taught BR96-Dox was more effective at killing tumors in vivo with less toxicity and more activity when compared to untargeted Dox (abstract).
Firestone taught conjugation of the monoclonal antibody to Dox by a hydrazone bond, wherein a hydrazone bond is reasonably stable at neutral pH, but released rapidly within lysosomes, where the pH is about 5 (page 252, right column, first paragraph).
Firestone taught BR96 DOX comprised a linker of 6-Maleimidocaproylhydrazone (page 253, left column, second paragraph).
Marciani taught conjugation of moieties to a saponin wherein the conjugation site is an aldehyde at position C23 of the aglycone core (pages 23-25, Schemes 5a-b).
Regarding instant claims 57-59, 61, 64-65, 67-68, 70-75, and 77-78, it would have been obvious for a person having ordinary skill in the art to take the effective method of cancer treatment of a subject by silencing a protein in a cancer cell comprising administering a pharmaceutical combination comprising a single pharmaceutical composition:
a first pharmaceutical composition comprising a first conjugate comprising an EGFR targeted antibody covalently linked to an effector molecule wherein the effector molecule is an siRNA oligonucleotide; and
a second pharmaceutical composition comprising a second conjugate comprising an EGFR targeted antibody covalently linked to melittin which is an endosomolytic moiety that causes endosomal escape agent
of Geall – and:
Exchange the EGFR targeting moieties with a bivalent CD71 targeted single domain antibody, wherein the bivalent antibodies are the same in view of Geall, Niewoehner, and Heukers;
Exchange the endosomal escape molecule melittin for the endosomal escape molecule glycoside saponin SO1861 of Weng;
Exchange the attachment linker of the endosomal escape moiety for an acid sensitive hydrazone linker of Firestone for linking the payload;
covalently conjugate the acid sensitive hydrazone linker that releases the SO1861 payload to cause endosomal escape at the C23 aldehyde of SO1861 as taught by Marciani.
This is obvious because:
i) Geall taught the antibody as targeting CD71 with an antibody siRNA conjugate effectively and as a sdAb; ii) Heukers taught a bivalent biparatopic EGFR-targeted single domain antibody 7D12-9G8 was effective and the VHH causes faster accumulation at the tumor, a more homogeneous distribution within the tumor, and a more rapid clearance of unbound molecules, compared to an anti-EGFR monoclonal antibody and is known to go through endosomal clearance and kill cancer cells; and iii) Niewoehner taught a bivalent transferrin receptor (TfR or CD71) antibody leads to effective lysosomal targeting,wherein the bivalent TfR antibody is the same. Thus, a bivalent CD71 sdAb would be obvious and expected to be effective;
Weng taught the saponin SO1861: i) augments the escape of the genetic cargo out of the intracellular compartments into the cytosol; ii) integrated into a receptor targeted nanoplex improved DNA transfection and provides improved effector properties; and iii) in combination with siRNA increased siRNA silencing in cancer cells;
Firestone taught BR96 bound to tumor cells, then internalized via endocytosis into lysosomes, wherein the acidic milieu of the lysozymes hydrolyzed the hydrazone linkage, releasing free payload effectively into the cancer cells. Firestone taught conjugation of the monoclonal antibody to Dox by a hydrazone bond, wherein a hydrazone bond is stable at neutral pH, but released rapidly within lysosomes. Further, Weng taught SO1861 induces the release of the genetic cargo out of endosomes and lysosomes, wherein the lumen of the endosomes and lysosomes are acidified. Thus, the hydrazone linker would release the SO1861 payload in the endosomes/lysosomes, where SO1861 has its effects; and
Marciani taught conjugation of moieties to a saponin wherein the conjugation site is an aldehyde at position C23 of the aglycone core.
There is a reasonable expectation of success because:
i) Geall taught the antibody as targeting CD71 with an antibody siRNA conjugate effectively; ii) Heukers taught a bivalent biparatopic EGFR-targeted single domain antibody 7D12-9G8 was effective and the VHH causes faster accumulation at the tumor, a more homogeneous distribution within the tumor, and a more rapid clearance of unbound molecules, compared to an anti-EGFR monoclonal antibody and is known to go through endosomal clearance and kill cancer cells; and iii) Niewoehner taught a bivalent transferrin receptor (TfR or CD71) antibody leads to effective lysosomal targeting, wherein the bivalent TfR antibody is the same. Thus, a bivalent CD71 VHH sdAb would be obvious and expected to be effective;
The saponin SO1861 would: i) augment the escape of the siRNA genetic cargo out of the intracellular compartments into the cytosol; ii) improve DNA transfection and provide improved effector properties; and iii) increase siRNA silencing in cancer cells to promote;
Hydrazone linkers are known to be effective, wherein hydrazone linkers are stable a neutral pH of 7, but once internalized into the acidic conditions, the payload is released into cancer cells. Further, Weng taught SO1861 induces the release of the genetic cargo out of endosomes and lysosomes, wherein the lumen of the endosomes and lysosomes are acidified. Thus, the hydrazone linker would release the SO1861 payload(s) in the endosomes/lysosomes, where SO1861 has its effects; and
an acid sensitive hydrazone linker connected at the C23 aldehyde of SO1861 would connect to the carbonyls and release the payload under acidic conditions to allow the SO1861 to perform the endosomal escape of the siRNA.
This would produce a method of cancer treatment of a subject by silencing a protein in a cancer cell comprising administering a single pharmaceutical combination (instant claim 59) comprising two pharmaceutical compositions (instant claim 58) comprising:
a first pharmaceutical composition a first conjugate comprising a VHH single domain CD71 targeted antibody, wherein the bivalent TfR VHH antibody is the same (instant claim 61), covalently linked to an effector molecule wherein the effector molecule is an siRNA oligonucleotide (instant claims 64-65); and
a second pharmaceutical composition comprising a second conjugate comprising a single domain CD71 targeted VHH antibody wherein the bivalent TfR VHH antibody is the same, covalently linked to a linker and a modified SO1861 (instant claims 71-74) which is a bidesmosidic triterpene glycoside with an aglycone core structure of quillaic acid with a first saccharide chain of Gal-(1-2)-[Xyl-(1-3)]-GlcA- (instant claim 68), wherein the first saccharide chain would comprise a the glucuronic acid unit comprising a carboxyl group (instant claim 70), and wherein the SO1861 linker comprises an acid sensitive linker comprising a hydrazone bond linker connected at the C23 aldehyde of SO1861 (instant claims 67 and 77-78) which breaks under acidic conditions such that it forms an aldehyde function at C23 of the quillaic aglycone core structure to release the payload to cause endosomal escape (instant claims 57 and 75).
Regarding instant claim 76, it would have been obvious for a person having ordinary skill in the art to take the composition from the method of Geall, Heukers, Niewoehner, Weng, Firestone, and Marciani above of a pharmaceutical composition comprising two pharmaceutical compositions comprising:
a first pharmaceutical composition a first conjugate comprising a single domain CD71 targeted VHH antibody covalently linked to an effector molecule wherein the effector molecule is an siRNA oligonucleotide; and
a second pharmaceutical composition comprising a second conjugate comprising a single domain CD71 targeted VHH antibody covalently linked to a linker and a modified SO1861 which is a bidesmosidic triterpene glycoside with an aglycone core structure of quillaic acid with a first saccharide chain of Gal-(1-2)-[Xyl-(1-3)]-GlcA wherein the linker comprises an acid sensitive linker comprising a hydrazone bond linker connected at the C23 aldehyde of SO1861 which breaks under acidic conditions such that it forms an aldehyde function at C23 of the quillaic aglycone core structure to release the payload to cause endosomal escape
– and: include the compositions in the effective in vitro method for transferring an oligonucleotide effector molecule from outside a CD71 expressing cell to inside of said cell of Weng;
This is obvious because: 1) the method of Weng taught SO1861 was effective at transferring oligonucleotides into cells when targeted to the cell, Thus, the two compositions comprising SO1861 or an siRNA effector could be used for transferring nucleotides into the cell.
There is a reasonable expectation of success because: 1) SO1861 is a known endosomal escape agent that can effectively increase the effectiveness of oligonucleotide effector into the cell and the VHH would target the agents to the cells for internalization to the endosome..
This would produce an in vitro method for transferring an oligonucleotide effector molecule from outside an CD71 expressing cell to inside of said cell comprising:
providing a pharmaceutical composition comprising a first pharmaceutical composition a first conjugate comprising a bivalent single domain CD71 targeted VHH antibody covalently linked to an effector molecule wherein the effector molecule is an siRNA oligonucleotide;
providing a pharmaceutical composition comprising a second pharmaceutical composition of a second conjugate comprising a bivalent single domain CD71 targeted VHH antibody covalently linked to a linker and a modified SO1861 wherein the linker comprises an acid sensitive linker comprising a hydrazone bond linker connected at the C23 aldehyde of SO1861 which breaks under acidic conditions such that it forms an aldehyde function at C23 of the quillaic aglycone core structure to release the payload to cause endosomal escape;
providing a cell that expresses CD71 on the surface wherein the surface of the cell is targeted by the pharmaceutical compositions in a) and b);
contacting the cell of step c) in vitro with the pharmaceutical composition of step a) and the pharmaceutical composition of step b).
therewith establishing the transfer of the oligonucleotide effector from outside the cell into the cell.
Response to Arguments
Applicant has amended the independent claim 57.
The updated to rejection is above.
Applicant argues the cited documents fail to teach a conjugate comprising a single-domain antibody is a VHH domain that can bind to CD71, wherein the first cell-surface molecule is CD71.
Applicant argues Geall fails to teach a VHH domain that can bind to CD71. Geall discloses EGFR antibody ligand conjugates, which are conjugated to siRNA, (see [0689] and FIG. 23).
In addition, Geall fails to teach a pharmaceutical combination of a first conjugate VHH domain that can bind to CD71 and a modified saponin.
The Examiner suggests that because Gaell would teach second conjugate comprising an EGFR targeted antibody covalently linked to melittin this would render it obvious to use another endosomal escape molecule. Applicant respectfully disagrees.
Applicant argues as acknowledged by the Examiner, Melittin is not a endosomolytic small molecule, but is a 26-membered endosomolytic polypeptide. Although [0389] mentions the use of endosomolytic small molecule as endosomolytic moiety in general, it is described in the context of a molecule of Formula (1), which is A-X-B-Y-C, which may further comprise an additional conjugating moiety. The molecule contains A, which is a binding moiety, B, which is a polynucleotide, and C, which is a polymer. In some instances, the additional conjugating moiety is an endosomolytic moiety and this endosomolytic moiety may be selected to be a endosomolytic small molecule. That means that the conjugate of Formula (1), besides the endosomolytic moiety, also contains a polynucleotide and a polymer. This is different from the claimed subject matter, wherein the first conjugate contains the polynucleotide, while the modified saponin is not part of said first conjugate.
Applicant argues thus, Geall merely teaches a single conjugate comprising a binding moiety conjugated to a polynucleic acid molecule and a polymer, see, e.g., Geall at [0004]. In particular, Geall teaches an antibody-siRNA-PEG conjugate, where a variety of antibodies can be conjugated to siRNAs with various PEG sizes or polymer linkers. The preferred antibodies of Geall are full-length antibodies such as zalutumumab, panitumumab, and herceptin, as well as other full-sized antibodies for PSMA, ASGR, and anti-B cell. Example 8 teaches that chol-siRNA conjugates cleared much faster from plasma than the mAb-siRNA conjugates. Thus, the preferred antibody is one that is large enough, e.g., - 150 kDa, that avoids rapid renal clearance. One of skill in the art would have no motivation to merely "exchange" the full-length antibody with a singledomain antibody based on the disclosure of Geall even in view of the teachings ofHeukers.
Applicant further notes that:
- the Example related to FIG. 86 is seemingly based on a mouse anti-B cell antibody (and not on an anti-B cell Fab) conjugated to an siRNA designed to down regulate HPRT mRNA. The anti-B cell antibody is not specified;
- the CD71-Fab, as indicated in FIG. 54, is not an a CD71 targeted sdAb; and
- the example of FIG. 86 does not show the use of melittin or any other conjugate having an endosomolytic moiety and that the only examples showing the use of a conjugate of melittin is based on EGFR ligands.
Applicant argues therefore, one of skill in the art would have no motivation to a combination of CD7 l targeted ligand in combination with the melittin endosomolytic moiety within one conjugate.
Further, as acknowledged in the Office Action, Geall fails to disclose the modified saponin as claimed. The Office Action suggests that Geall would teach that melittin is an endosomolytic moiety that causes endosomal escape agent [0689]. Example 8, to which the Office is referring to, evaluates whether the addition of melittin increased delivery of the siRNA to the target tissue (passage copied here):
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Applicant argues the results showed that adding melittin slightly reduced tumor uptake. The results of Example 8 teach away from at least administering a second compound or conjugate to increase tumor uptake of the first conjugate through the addition of an endosomal escape peptide, such as melittin. The Office must consider the prior art in its entirety. MPEP § 2141.02. One of skill in the art evaluating the cited reference as a whole would not be motivated to (a) add a second conjugate and (b) add a second conjugate with an endosomal escape enhancer.
Applicant argues Heuskers cannot remedy these deficiencies. Heuskers fails to teach an conjugate having a single-domain antibody, which is a VHH domain and that can bind to CD71. Heuker's teaching is specific nanobodies targeting the epidermal growth factor receptor (EGFR) and is used to support photodynamic therapy of a photosensitizer. There is no motivation to apply the teaching of Heuker to Geall and there is not disclosure of sdAb for targeting CD71.
Applicant argues furthermore, Heukers states "[i]mportantly, free PS, R2-PS, and the non-illuminated EGFR-targeted NB-PS conjugates did not affect cell viability ... which highlights the specificity of the PDT approach." (See Heukers at 1443). One of skill in the art would have no motivation nor any reasonable expectation of success to substitute the antibody fragments of Heukers for the full-length antibodies of Geall.
Applicant argues Weng cannot remedy these deficiencies. The Office Action suggests that it would not be obvious to substitute the polypeptide melittin of Geall with SO 1861 as disclosed in Weng. Applicant respectfully submits that the rejection does not establish an articulated reason to make this substitution with a reasonable expectation of success. Geall teaches melittin, a linear peptide of 26 amino acids, which has a structure as follows:
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In contrast, the pending claims relate to a saponin having a gypsogenin core or quillaic acid core with the following general structure:
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Applicant argues as referenced above Geall dissuades a person of skill in the art from using melittin because the melittin did not increase tumor uptake, rather there was an increase in plasma clearance of the conjugate. One of skill in the art would not look to Weng to cure the deficiencies of Geall.
Applicant argues Weng's teaching is specific to peptide- and lipid-based nanoplexes. Weng reports delivery enhancement when SO1861 is used in targeted nanoplexes (e.g., peptide/DNA complexes and lipid-containing nanoplexes). The claimed pharmaceutical composition, by contrast, concerns a ligand/effector conjugate administered as such, not as a cationic peptide/lipid nanoplex.
Applicant argues there is no reasonable expectation to observe an enhancement in nanoplex formulations to translate to a materially different delivery format involving soluble antibody conjugates of siRNA. Weng' s hypothesized mechanism is also formulation-dependent, namely that SO 1861 associates with the positively charged nanoplexes (LPDS) and then mediates endosomal escape (see page 88).
Applicant argues at most, Weng suggests that SO1861 can enhance delivery in nanoplex-based non-viral transfection systems, see the abstract; it does not teach, suggest, or provide a reasonable expectation that SO1861 would be an interchangeable substitute for melittin in the antibody conjugate setting of Geall.
Applicant argues a skilled person would not reasonably expect from Weng that the SO 1861, which is a small molecule, would be suitable for covalently coupling to an antibody conjugate of Geall according to Formula (1), how it would be incorporated, and, if it could have any substantive effect on endosomal escape when conjugated. As described by Weng the SO 1861 can be easily integrated into the lipid bilayer of glycerophospholipid model membranes, this is fundamentally different from covalently coupling to an antibody conjugate. The proposed substitution cannot change the principle of operation of Geall for its intended purpose.
Additionally, Weng proposes the mechanism that SO1861 first associates with the positively charged nanoplexes and mediates in a second step the endosomal escape of nanoplexes via its membrane active properties. So, these mechanisms do not provide a motivation or a reasonable expectation of success for the antibody-conjugate setting of Geall.
Applicant argues Firestone cannot remedy any of the deficiencies cited here above. Firestone describes BR96-Dox, which is an immunoconjugate in which doxorubicin is linked via an acid-labile hydrazone to the chimeric MAb BR96. The covalent bond of the doxorubicin in the conjugate is established by a hydrazone bond of the doxorubicin derivative.
Doxorubicin:
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The Office Action asserts that it would have been obvious to substitute the aldehyde group of the at C23 of the cyclic part of the aglycone core structure of SO1861 of Weng by a hydrazone bond as disclosed in Firestone. Applicant respectfully disagrees. Weng does not teach to modify the SO 1861 or form a conjugate containing SO 1861. In fact, the complexation of SO1861 with nanoplexes is a completely different approach. Additionally, while Firestone talks about off target toxicity of doxorubicin, Weng does not have such an issue for example as SO1861 is complexed within the nanoplexes. The proposed substitution cannot change the principle of operation of Weng for its intended purpose.
Secondly, Applicant submits that the proposed modification as suggested in the Office Action is not chemically feasible. Firestone does not provide any chemical pathways to form such a hydrazone bond and merely mentions that a Dox 3-(2-pyridyldithio)propionyl hydrazone and Dox 6-maleimidocaproylhydrazone is commercially available. Lastly, Firestone is also not relevant for targeted saponin delivery as doxorubicin is not a saponin. Firestone fails to teach targeting of a saponin.
Applicant argues Marciani cannot remedy these deficiencies, Marciani is directed to triterpene saponin analogs having adjuvant and immunostimulatory activity, with success metrics relating to vaccine/adjuvant design (immune response enhancement, purification, stability, toxicity profile in that context). Applicants' claimed combination is not directed to vaccine adjuvanticity or immunostimulant, but to delivery/therapeutic action of an antibody-nucleic acid conjugate for treating a cancer by silencing genes (gene suppression). In this therapeutic setting, increasing immunostimulatory activity is not an objective and would be undesirable.
Further, Marciani's "success" relates to adjuvanticity and immunostimulation, not improving intracellular delivery of antibody-effector conjugates. The Office Action does not identify a teaching in Marciani that derivatization at the aldehyde group of aglycone core structure would predictably preserve, let alone improve, delivery performance in the Geall context, nor does it explain why a modification developed for vaccine adjuvant properties would beneficially impact Ab-nucleic acid mediated delivery.
Applicant argues, the rejection improperly assumes that because Marciani identifies a conjugation site and reports advantages in a vaccine-adjuvant context, the same modification would have been predictably advantageous in a distinct therapeutic delivery context.
Applicant argues in addition, Marciani is also not relevant for targeted saponin delivery wherein the saponin is a conjugate comprising an Ab-ligand. Thus, Marciani does not proper provide a motivation for features of any one of the claims to make any of these modification in the present context and does not provide a reasonable expectation of success.
In response, Applicant's arguments filed 2/10/2026 have been fully considered
but they are not persuasive. Independent claim 57 has been amended to require a conjugate comprising a single-domain antibody is a VHH domain that can bind to CD71, wherein the first cell-surface molecule is CD71 and the updated rejection is above.
Regarding Geall and melittin,
As described above, Geall taught an effective method of cancer treatment by silencing a protein in a cancer cell comprising administering a single pharmaceutical combination comprising:
a first conjugate comprising an EGFR targeted antibody covalently linked to an effector molecule wherein the effector molecule is an siRNA oligonucleotide; and
a second conjugate comprising an EGFR targeted antibody covalently linked to melittin (page 148, [0689]),
wherein melittin is an endosomolytic moiety that causes endosomal escape agent (page 148, [0689]), wherein the expression of KRAS in the tumor was effectively silenced from 24 to 168 hours and wherein KRAS was decreased more in tumors that were administered the combination compared to an EGFR targeted antibody covalently linked to the siRNA alone (Fig. 33). Geall further taught: a) the antibody as a single-domain antibody (sdAb) (page 1, [0019]); b) the endosomolytic moiety as an endosomolytic small molecule and an endosomolytic polymer (page 73, [0389]). Thus, targeting an siRNA oligonucleotide conjugated to a sdAb in combination with an endosomolytic moiety that causes endosomal escape agent conjugated to a sdAb would be obvious with a reasonable expectation of success.
Additionally, biparatopic VHH targeted antibodies are known to be effective and the VHH causes faster accumulation at the tumor, a more homogeneous distribution within the tumor, and a more rapid clearance of unbound molecules, compared to an anti-EGFR monoclonal antibody and is known to go through endosomal clearance and kill cancer cells.
Further, as described above, Niewoehner taught a bivalent transferrin receptor (TfR or CD71) antibody leads to effective lysosomal targeting, wherein the bivalent TfR antibody is the same (abstract and Fig. 2E). Thus, a bivalent CD71 targeted VHH conjugate would be obvious and expected go through endosomal clearance.
Thus, targeting an siRNA oligonucleotide conjugated to a bivalent CD71 targeted VHH conjugate in combination with an endosomolytic moiety that causes endosomal escape agent conjugated to a bivalent CD71 targeted VHH conjugate would be obvious with a reasonable expectation of success.
Regarding Geall and a PEG linker, combination treatment with EGFR antibody-KRAS-PEG5k and EGFR targeted antibody-melittin was shown to be effective in a tumor in vivo (Fig. 33). The EGFR antibody-KRAS-PEG5k contains a PEG linker connected at the end of the siRNA, not between the antibody and the siRNA. Thus, the antibody to siRNA linker does not involve PEG and does not change the principle of operation of Geall. Further, the claim does not exclude siRNA modifications attached to the end of the siRNA and antibody siRNA conjugates without PEG attachments are functional. Regarding Geall Fig. 54 and 86, Geall Fig. 54 indicates that HPRT was tested in B cells with a CD71 (Fab) and Fig. 86 identifies the HPRT measured in mouse B cells (Geall, page 221, [1107]). Thus, Geall taught effectively targeting CD71 for siRNA delivery. Regarding chol-siRNA clearance and a 150kDa antibody, biparatopic VHH targeted antibodies are known to be effective and VHH are known to cause faster accumulation at the tumor and a more homogeneous distribution within the tumor compared to an anti-EGFR monoclonal antibody and is known to go through endosomal clearance and kill cancer cells. Thus, a full-length antibody is not required. Therefore, targeting an siRNA oligonucleotide conjugated to a bivalent CD71 targeted VHH conjugate in combination with an endosomolytic moiety that causes endosomal escape agent conjugated to a bivalent CD71 targeted VHH conjugate would be obvious with a reasonable expectation of success.
Regarding the saponin, Fig. 33 shows combination treatment with EGFR antibody-KRAS-PEG5k and EGFR targeted antibody-melittin was shown to be effective in a tumor in vivo. Including EGFR targeted antibody-melittin in the composition caused tumor KRAS mRNA to be lower at all time points tested in Fig. 33.. Thus, including EGFR targeted antibody-melittin in the composition was more effective at lowering target mRNA levels, not less effective, in vivo. These results do not teach away from including an endosomolytic moiety that causes endosomal escape agent conjugated to an antibody or sdAb.
Regarding substitution with a reasonable expectation of success of melittin for SO1861 and Weng – Geall taught melittin is an endosomolytic moiety that causes endosomal escape agent (page 148, [0689]) and Weng taught to exert a silencing effect, siRNA has to escape from the endosomes/lysosomes into the cytosol because in lysosomes siRNA/DNA are degraded by nucleases (page 75-76 bridging paragraph). Weng taught: 1) If the endo-/lysosomal escape of the genetic cargo does not occur the siRNA/DNA is degraded within the lysosome thus leading to a therapeutic failure; 2) limited release of siRNA/DNA into the cytosol is one of the major obstacles for efficient siRNA/DNA delivery; and 3) development of innovative strategies to augment the endo-/lysosomal escape of siRNA/DNA and the delivery of a minimal amount of siRNA/DNA are desired for an efficient therapeutic response (page 75-76 bridging paragraph). Weng taught the saponin SO1861: i) augments the escape of the genetic cargo out of the intracellular compartments into the cytosol; ii) integrated into a receptor targeted nanoplex improved DNA transfection and provides improved effector properties; and iii) in combination with siRNA increased siRNA silencing in cancer cells. Thus, exchanging one effective endosomal escape agent for another is obvious with a reasonable expectation of success. The covalent bond to SO1861 would be a hydrazone linker as described above. Weng taught SO1861 induces the release of the genetic cargo out of endosomes and lysosomes, wherein the lumen of the endosomes and lysosomes are acidified.
Regarding Weng, SO1861 nanoplexes, SO1861 suitability for covalently coupling to an antibody, and Firestone:
Geall taught a second conjugate comprising an antibody covalently linked to melittin (page 148, [0689]), wherein melittin is an endosomolytic moiety that causes endosomal escape agent (page 148, [0689]). As described above, Weng taught SO1861 induces the release of the genetic cargo out of endosomes and lysosomes, wherein the lumen of the endosomes and lysosomes are acidified. Additionally, as described above, Marciani taught conjugation of moieties to a saponin wherein the conjugation site is an aldehyde at position C23 of the aglycone core (pages 23-25, Schemes 5a-b) and Firestone taught an acid-labile hydrazone conjugated to the chimeric MAb BR96 (abstract), wherein internalization of the antibody conjugate via endocytosis into lysosomes led to the acidic milieu of the lysozymes hydrolyzing the hydrazone linkage, releasing free Dox payload effectively (abstract). Thus, a hydrazone linker would release the SO1861 payload form the antibody in the endosomes/lysosomes, where SO1861 has its effects. Thus, conjugation of an VHH antibody to an endosomal escape unit of SO1861 via a hydrazone linker is obvious with a reasonable expectation of success.
Regarding the chemical feasibility and relevance of a hydrazone bond, the hydrazone bond allows for conjugation of a payload to an antibody with acidic release within the lysosomal pathway. Further, Geall taught an acid cleavable linker as a hydrazone linker, which is susceptible to hydrolytic cleavage (Geall, page 82, [0507]). Thus, it is within the skill of a person having ordinary skill in the art to utilize a hydrazone linker in an antibody conjugate.
Regarding Marciani, Marciani taught conjugation of moieties to a saponin wherein the conjugation site is an aldehyde at position C23 of the aglycone core (pages 23-25, Schemes 5a-b). Thus, conjugation of saponins at C23 would be obvious. A hydrazone bond at C23 would release the SO1861 molecule from an antibody in the endosomes/lysosomes, where SO1861 has its effect as described above and have a reasonable expectation of success.
Claims 57-59, 61, 64-65, and 67-78 are rejected under 35 U.S.C. 103 as being unpatentable over US 2017/0281795 (Geall AJ et al. reference of record), Heukers R et al. (Nanomedicine 2014 10(7):1441-51 reference of record), Niewoehner J et al. (Neuron 2014 81(1):49-60. doi: 10.1016/j.neuron.2013.10.061.) Weng A et al. (Journal of Controlled Release 2015 206 75-90 reference of record), Firestone RA e al. (Journal of Controlled Release 1996 39 (2-3) 251-259 reference of record), and US 2004/0242502 (Marciani D et al. reference of record) as applied to claims 57-59, 61, 64-65, 67-68, and 70-78 above, and further in view of Thermo Scientific et al. Instructions BMPH, EMCH, KMUH (https://documents.thermofisher.cn/TFS-Assets%2FLSG%2Fmanuals%2FMAN0011363_BMPH_EMCH_KMUH_UG.pdf 2015 reference of record).
Geall, Heukers, Niewoehner, Weng, Firestone, and Marciani are described above.
Geall did not teach conjugation of the endosomal escape moiety at a C23 of an aglycone core through reaction with EMCH, but this is obvious in view of Thermo.
Thermo taught EMCH (N-[ε-Maleimidocaproic acid) hydrazide is a heterobifunctional crosslinkers containing sulfhydryl-reactive maleimide and carbonyl-reactive hydrazide moieties, wherein hydrazide groups react with carbonyls (aldehydes and ketones) to form stable hydrazone bonds, wherein the EMCH crosslinker is useful for conjugating sulfhydryl-containing proteins (page 1, Introduction).
Regarding instant claim 69, it would have been obvious for a person having ordinary skill in the art to take the method of Geall, Heukers, Niewoehner, Weng, Firestone, and Marciani above and in the pharmaceutical composition: – 1) covalently conjugate the SO1861 payload with EMCH at an aldehyde of SO1861 as taught by Thermo, which would be the C23 aldehyde of SO1861.
This is obvious because: 1i) Thermo taught EMCH (N-[ε-Maleimidocaproic acid) hydrazide is a heterobifunctional crosslinkers containing sulfhydryl-reactive maleimide and carbonyl-reactive hydrazide moieties, wherein hydrazide groups react with carbonyls (aldehydes and ketones) to form stable hydrazone bonds, wherein the EMCH crosslinker is useful for conjugating sulfhydryl-containing proteins; and 1ii) the structure of EMCH is included in the 6-Maleimidocaproic acid hydrazone linker of Firestone.
There is a reasonable expectation of success because: 1i) Thermo taught EMCH (N-[ε-Maleimidocaproic acid) hydrazide is a heterobifunctional crosslinkers containing sulfhydryl-reactive maleimide and carbonyl-reactive hydrazide moieties, wherein hydrazide groups react with carbonyls (aldehydes and ketones) to form stable hydrazone bonds, wherein the EMCH crosslinker is useful for conjugating sulfhydryl-containing proteins; and 1ii) the structure of EMCH is included in the 6-Maleimidocaproic acid hydrazone linker of Firestone. Thus, the acid sensitive hydrazone EMCH linker connected at the C23 aldehyde of SO1861 would connect to the carbonyl and release the payload under acidic conditions to allow the SO1861 to perform the endosomal escape of the siRNA.
This would produce a method of cancer treatment of a subject by silencing a protein in a cancer cell comprising administering a single pharmaceutical combination comprising two pharmaceutical compositions comprising:
a first pharmaceutical composition a first conjugate comprising a VHH single domain CD71 targeted antibody, wherein the bivalent TfR VHH antibody is the same, covalently linked to an effector molecule wherein the effector molecule is an siRNA oligonucleotide; and
a second pharmaceutical composition of a second conjugate comprising a VHH single domain CD71 targeted antibody, wherein the bivalent TfR VHH antibody is the same, covalently linked to an EMCH linker and a modified SO1861 wherein the linker comprises an acid sensitive EMCH hydrazone bond linker connected at the C23 aldehyde of SO1861 which breaks under acidic conditions such that it forms an aldehyde function at C23 of the quillaic aglycone core structure to release the payload to cause endosomal escape.
Response to Arguments
Applicant has amended the independent claim 57.
The updated to rejection is above.
Applicant argues for the sake of completeness, it is recognized that ThermoFisher was also cited by the Official Action but solely with respect to features set forth in dependent claims. ThermoFisher does not remedy the aforementioned deficiencies.
In response, Applicant's arguments filed 2/10/2026 have been fully considered
but they are not persuasive. Independent claim 57 has been amended to require a conjugate comprising a single-domain antibody is a VHH domain that can bind to CD71, wherein the first cell-surface molecule is CD71 and the updated rejection is above. The teachings of ThermoFisher and the obvious rational is above.
Double Patenting
The nonstatutory double patenting rejection is based on a judicially created doctrine grounded in public policy (a policy reflected in the statute) so as to prevent the unjustified or improper timewise extension of the “right to exclude” granted by a patent and to prevent possible harassment by multiple assignees. A nonstatutory double patenting rejection is appropriate where the conflicting claims are not identical, but at least one examined application claim is not patentably distinct from the reference claim(s) because the examined application claim is either anticipated by, or would have been obvious over, the reference claim(s). See, e.g., In re Berg, 140 F.3d 1428, 46 USPQ2d 1226 (Fed. Cir. 1998); In re Goodman, 11 F.3d 1046, 29 USPQ2d 2010 (Fed. Cir. 1993); In re Longi, 759 F.2d 887, 225 USPQ 645 (Fed. Cir. 1985); In re Van Ornum, 686 F.2d 937, 214 USPQ 761 (CCPA 1982); In re Vogel, 422 F.2d 438, 164 USPQ 619 (CCPA 1970); In re Thorington, 418 F.2d 528, 163 USPQ 644 (CCPA 1969).
A timely filed terminal disclaimer in compliance with 37 CFR 1.321(c) or 1.321(d) may be used to overcome an actual or provisional rejection based on nonstatutory double patenting provided the reference application or patent either is shown to be commonly owned with the examined application, or claims an invention made as a result of activities undertaken within the scope of a joint research agreement. See MPEP § 717.02 for applications subject to examination under the first inventor to file provisions of the AIA as explained in MPEP § 2159. See MPEP § 2146 et seq. for applications not subject to examination under the first inventor to file provisions of the AIA . A terminal disclaimer must be signed in compliance with 37 CFR 1.321(b).
The filing of a terminal disclaimer by itself is not a complete reply to a nonstatutory double patenting (NSDP) rejection. A complete reply requires that the terminal disclaimer be accompanied by a reply requesting reconsideration of the prior Office action. Even where the NSDP rejection is provisional the reply must be complete. See MPEP § 804, subsection I.B.1. For a reply to a non-final Office action, see 37 CFR 1.111(a). For a reply to final Office action, see 37 CFR 1.113(c). A request for reconsideration while not provided for in 37 CFR 1.113(c) may be filed after final for consideration. See MPEP §§ 706.07(e) and 714.13.
The USPTO Internet website contains terminal disclaimer forms which may be used. Please visit www.uspto.gov/patent/patents-forms. The actual filing date of the application in which the form is filed determines what form (e.g., PTO/SB/25, PTO/SB/26, PTO/AIA /25, or PTO/AIA /26) should be used. A web-based eTerminal Disclaimer may be filled out completely online using web-screens. An eTerminal Disclaimer that meets all requirements is auto-processed and approved immediately upon submission. For more information about eTerminal Disclaimers, refer to www.uspto.gov/patents/apply/applying-online/eterminal-disclaimer.
Claims 57-58 are provisionally rejected on the ground of nonstatutory double patenting as being unpatentable over claims 41-57 of copending Application No.19/044,661. Although the claims at issue are not identical, they are not patentably distinct from each other because:
Regarding instant claims 57-58, the copending claims taught a method for treating a disease condition associated with the presence of an aberrant cell, comprising administering to a human subject a therapeutic combination, wherein the therapeutic combination comprises:
(a) a first pharmaceutical composition comprising a first proteinaceous molecule comprising a first binding site for binding to a first cell-surface molecule of the sdAb VHH and at least one saponin covalently bound to said first proteinaceous molecule preferably covalently bound to an amino- acid residue of said first proteinaceous molecule, the first pharmaceutical composition optionally further comprising a pharmaceutically acceptable excipient; and
(b) a second pharmaceutical composition comprising a second proteinaceous molecule, the second proteinaceous molecule comprising a second binding site for binding to a second cell-surface molecule different from the first cell-surface molecule as a sdAb VHH and an oligonucleotide effector moiety, the second pharmaceutical composition optionally further comprising a pharmaceutically acceptable excipient,
wherein the at least one saponin is a bisdesmosidic triterpene saponin belonging to the type of a 12,13-dehydrooleanane with an aldehyde function in position C-23, wherein the saponin is covalently coupled the first proteinaceous molecule, preferably covalently coupled to an amino- acid residue of the first proteinaceous molecule, via an aldehyde function in the saponin, preferably said aldehyde function in position C-23, preferably via at least one linker, and/or via at least one cleavable linker,
wherein the amino-acid residue preferably is selected from cysteine and lysine in copending claim 41, and wherein the first binding sire and second binding site comprise an antibody that targeted CD71 in copending claim 45 (instant claims 57-58).
This is a provisional nonstatutory double patenting rejection because the patentably indistinct claims have not in fact been patented.
Response to Arguments
Applicant has amended the independent claim 57.
The updated to rejection is above.
Applicant requests that the provisionally rejection of the claims under the judicially created doctrine of obviousness-type double patenting as allegedly being unpatentable over the claims of co-pending applications Nos. 19/044,661, 17/312, 104, 17/312,476, 17/415,759, 17/312,019, 18/012,741, 18/012,723, 18/012,729, 18/012,760, 17/312,193 and 18/723,163 be held in abeyance until indication by the Office of allowable claims in one of the co-pending applications.
In response, Applicant's arguments filed 2/10/2026 have been fully considered
but they are not persuasive. Independent claim 57 has been amended to require a conjugate comprising a single-domain antibody is a VHH domain that can bind to CD71, wherein the first cell-surface molecule is CD71 and the updated rejection is above.
A request to hold a rejection in abeyance is not a proper response to a rejection. Rather, a request to hold a matter in abeyance may only be made in response to an OBJECTION or REQUIREMENTS AS TO FORM (see 37 CFR 1.111(b) and MPEP §714.02).
Claims 57-59, 64-65, 67-74, and 77-78 are provisionally rejected on the ground of nonstatutory double patenting as being unpatentable over claims 41-57 of copending Application No.19/044,661.
The claims of copending ‘661 teach the limitations of claims 57-58 for the reasons set forth above.
The claims of ‘661 are described above.
The claims of ‘661 further taught regarding instant claims 57-59, 64-65, 67-74, and 77-78, the copending claims taught a method for treating a disease condition associated with the presence of an aberrant cell, comprising administering to a human subject a therapeutic combination, wherein the therapeutic combination comprises:
(a) a first pharmaceutical composition comprising a first proteinaceous molecule comprising a first binding site for binding to a first cell-surface molecule of the sdAb VHH and at least one saponin covalently bound to said first proteinaceous molecule preferably covalently bound to an amino- acid residue of said first proteinaceous molecule, the first pharmaceutical composition optionally further comprising a pharmaceutically acceptable excipient; and
(b) a second pharmaceutical composition comprising a second proteinaceous molecule preferably different from the first proteinaceous molecule, the second proteinaceous molecule comprising a second binding site for binding to a second cell-surface molecule different from the first cell-surface molecule as a sdAb VHH and an oligonucleotide effector moiety, the second pharmaceutical composition optionally further comprising a pharmaceutically acceptable excipient,
wherein the at least one saponin is a bisdesmosidic triterpene saponin belonging to the type of a 12,13-dehydrooleanane with an aldehyde function in position C-23, wherein the saponin is covalently coupled the first proteinaceous molecule, preferably covalently coupled to an amino- acid residue of the first proteinaceous molecule, via an aldehyde function in the saponin, preferably said aldehyde function in position C-23, preferably via at least one linker, and/or via at least one cleavable linker,
wherein the amino-acid residue preferably is selected from cysteine and lysine in copending claim 41,
wherein the saponin is SO1861 in copending claim 43,
wherein the aldehyde function in position C-23 of the at least one saponin is covalently coupled to linker N-e-maleimidocaproic acid hydrazide, which is an acid sensitive linker in copending claim 44,
wherein the first binding sire and second binding site comprise an antibody that targeted CD71 in copending claim 45,
wherein the effector moiety is an siRNA in copending claim 46,
wherein the first proteinaceous molecule comprises more than one covalently bound saponin via at least one oligomeric or polymeric scaffold in copending claim 50,
wherein the at least one saponin is a bisdesmosidic triterpene saponin belonging to the type of a 12,13-dehydrooleanane with an aldehyde function in position C-23 , wherein the saponin is covalently coupled the first proteinaceous molecule via an aldehyde function in the saponin in position C-23 via at least one cleavable linker, wherein the cleavable linker is subject to cleavage in vivo under acidic conditions as present in endosomes and/or lysosomes of mammalian cells, preferably human cells, preferably at pH 4.0 - 6.5, and more preferably at pH s; 5.5 in copending claim 53,
wherein the first pharmaceutical composition and the second pharmaceutical composition are administered to the patient in need thereof in copending claim 55,
A method for treating a disease condition associated with the presence of an aberrant cell, in a patient in need thereof comprising administering to the patient the first pharmaceutical composition, wherein the first pharmaceutical composition further comprises the second proteinaceous molecule in copending claim 56.
The claims of ‘661 did not teach a single embodiment of a first and second molecule as sdAb that targeted CD71, wherein the first and second conjugate are provided in a single pharmaceutical composition, with an siRNA as the effector, wherein the saponin is SO1861, wherein the C23 position is reacted with N-e-maleimidocaproic acid hydrazide, but this is obvious in view of copending claims 43-44, 46, 53, and 55.
Regarding instant claims 57-59, 64-65, 67-74, and 77-78, it would have been obvious for a person having ordinary skill in the art to take the pharmaceutical composition of: a) a first conjugate that comprised a VHH anti-CD71 conjugated to a saponin at C23; and b) a second conjugate that comprised a VHH anti-CD71 conjugated to an nucleic acid effector moiety of copending claims 41 and 45 – and: 1) use SO1861 as the saponin as taught in copending claim 43, covalently couple the linker N-e-maleimidocaproic acid hydrazide, which is an acid sensitive linker in copending claim 44, use siRNA as the effector moiety as taught in copending claim 46, wherein the saponin is covalently coupled the first proteinaceous molecule via an aldehyde function in the saponin in position C-23 via at least one cleavable linker, wherein the cleavable linker is subject to cleavage in vivo under acidic conditions as present in endosomes and/or lysosomes of mammalian cells, preferably human cells, preferably at pH 4.0 - 6.5, and more preferably at pH s; 5.5 in copending claim 53, and wherein the first pharmaceutical composition and the second pharmaceutical composition are administered to the patient in need thereof in copending claim 55.
This is obvious with a reasonable expectation of success because copending claims 43-44, 46, 53, and 55 taught these embodiments and 1) SO1861 is the saponin; 2) the linker N-e-maleimidocaproic acid hydrazide is an acid sensitive linker; 3) siRNA is an effector moiety that can decrease protein expression; 4) a saponin covalently coupled the first proteinaceous molecule via an aldehyde function in the saponin in position C-23 via at least one cleavable linker, wherein the cleavable linker is subject to cleavage in vivo under acidic conditions as present in endosomes and/or lysosomes of mammalian cells would release the SO1861 payload in the cells; and 5) administering the first and second pharmaceutical compositions would allow the diseased cells to be targeted with both the first and second compositions together.
This is a provisional nonstatutory double patenting rejection because the patentably indistinct claims have not in fact been patented.
Response to Arguments
Applicant has amended the independent claim 57.
The updated to rejection is above.
Applicant requests that the provisionally rejection of the claims under the judicially created doctrine of obviousness-type double patenting as allegedly being unpatentable over the claims of co-pending applications Nos. 19/044,661, 17/312, 104, 17/312,476, 17/415,759, 17/312,019, 18/012,741, 18/012,723, 18/012,729, 18/012,760, 17/312,193 and 18/723,163 be held in abeyance until indication by the Office of allowable claims in one of the co-pending applications.
In response, Applicant's arguments filed 2/10/2026 have been fully considered
but they are not persuasive. Independent claim 57 has been amended to require a conjugate comprising a single-domain antibody is a VHH domain that can bind to CD71, wherein the first cell-surface molecule is CD71 and the updated rejection is above.
A request to hold a rejection in abeyance is not a proper response to a rejection. Rather, a request to hold a matter in abeyance may only be made in response to an OBJECTION or REQUIREMENTS AS TO FORM (see 37 CFR 1.111(b) and MPEP §714.02).
Claims 57-59, 61, 64-65, 67-74, and 77-78 are provisionally rejected on the ground of nonstatutory double patenting as being unpatentable over claims 41-57 of copending Application No.19/044,661 and further in view of Heukers R et al. (Nanomedicine 2014 10(7):1441-51 reference of record) and Niewoehner J et al. (Neuron 2014 81(1):49-60. doi: 10.1016/j.neuron.2013.10.061.).
The claims of copending ‘661 teach the limitations of claims 57-59, 64-65, 67-74, and 77-78 for the reasons set forth above.
‘661 is described above.
‘661 did not teach the first conjugate comprises at least two sdAbs that were the same, but this is obvious in view of Heukers and Niewoehner.
Heukers taught EGFR-targeted nanobody® VHH single domain conjugates are specific and potently kill cancer cells (abstract and Fig. 2). Heukers taught a biparatopic EGFR-targeted single domain antibody 7D12-9G8 that is known to be internalized (page 1442, left column, first paragraph). Heukers taught 7D12-9G8-PS induces the lysosomal trafficking (page, 1448, right column first paragraph). Heukers taught EGFR-targeted single domain antibodies showed a faster accumulation at the tumor, a more homogeneous distribution within the tumor, and a more rapid clearance of unbound molecules, compared to an anti-EGFR monoclonal antibody (page 1442, left column, first paragraph).
Niewoehner taught a bivalent transferrin receptor (TfR or CD71) antibody leads to effective lysosomal targeting, wherein the bivalent TfR antibody is the same (abstract and Fig. 2E).
Regarding instant claim 61, it would have been obvious for a person having ordinary skill in the art to take the method of ‘661 above and in the pharmaceutical composition: – 1) exchange the targeting sdAb of the first and second conjugate with a monospecific bivalent CD71 single domain VHH antibody of Heukers and Niewoehner.
This is obvious because: 1a) Heukers taught a bivalent biparatopic EGFR-targeted single domain antibody 7D12-9G8 was effective and the VHH causes faster accumulation at the tumor, a more homogeneous distribution within the tumor, and a more rapid clearance of unbound molecules, compared to an anti-EGFR monoclonal antibody and is known to go through endosomal clearance and kill cancer cells; and 1b) Niewoehner taught a bivalent transferrin receptor (TfR or CD71) antibody leads to effective lysosomal targeting,wherein the bivalent TfR antibody is the same. Thus, a monospecific bivalent CD71 sdAb would be obvious and expected to be effective.
There is a reasonable expectation of success because: 1a) Heukers taught a bivalent biparatopic EGFR-targeted single domain antibody 7D12-9G8 was effective and the VHH causes faster accumulation at the tumor, a more homogeneous distribution within the tumor, and a more rapid clearance of unbound molecules, compared to an anti-EGFR monoclonal antibody and is known to go through endosomal clearance and kill cancer cells; and 1b) Niewoehner taught a bivalent transferrin receptor (TfR or CD71) antibody leads to effective lysosomal targeting,wherein the bivalent TfR antibody is the same. Thus, a monospecific bivalent CD71 sdAb would be obvious and expected to be effective.
This is a provisional nonstatutory double patenting rejection because the patentably indistinct claims have not in fact been patented.
Response to Arguments
Applicant has amended the independent claim 57.
The updated to rejection is above.
Applicant requests that the provisionally rejection of the claims under the judicially created doctrine of obviousness-type double patenting as allegedly being unpatentable over the claims of co-pending applications Nos. 19/044,661, 17/312, 104, 17/312,476, 17/415,759, 17/312,019, 18/012,741, 18/012,723, 18/012,729, 18/012,760, 17/312,193 and 18/723,163 be held in abeyance until indication by the Office of allowable claims in one of the co-pending applications.
In response, Applicant's arguments filed 2/10/2026 have been fully considered
but they are not persuasive. Independent claim 57 has been amended to require a conjugate comprising a single-domain antibody is a VHH domain that can bind to CD71, wherein the first cell-surface molecule is CD71 and the updated rejection is above.
A request to hold a rejection in abeyance is not a proper response to a rejection. Rather, a request to hold a matter in abeyance may only be made in response to an OBJECTION or REQUIREMENTS AS TO FORM (see 37 CFR 1.111(b) and MPEP §714.02).
Claims 57-59, 61, 64-65, 67-68, and 70-78 are provisionally rejected on the ground of nonstatutory double patenting as being unpatentable over:
claims 1-2, 5, 7, 11, 13-14, 17-19, 23-26, and 28-29 of copending Application No. 17/312,104;
claims 1-2, 5, 8, 10-11, 16-19, 23-26, 29-30, and 32 of copending Application No. 17/312,476;
claims 1, 5-6, 9, 12-13, 20-24, 29-31, 35, 38, and 43 of copending Application No. 17/415,759
claims 1-2, 5, 9, 11, 17, 19, 23-24, 26, 28-29, and 39-43 of copending Application No. 17/312,019
in view of US 2017/0281795 (Geall AJ et al. reference of record), Heukers R et al. (Nanomedicine 2014 10(7):1441-51 reference of record), Niewoehner J et al. (Neuron 2014 81(1):49-60. doi: 10.1016/j.neuron.2013.10.061.), Weng A et al. (Journal of Controlled Release 2015 206 75-90 reference of record), and Firestone RA e al. (Journal of Controlled Release 1996 39 (2-3) 251-259 reference of record), and US 2004/0242502 (Marciani D et al. reference of record).
‘104 copending claims 1-2, 5, 7, 11, 13-14, 17-19, 23-26, and 28-29 claim an oligonucleotide covalently conjugated to at least one saponin with a hydrazone linker, wherein the saponin is a bisdesomsidic triterpene with an aldehyde in position C-23:
‘104 taught an oligonucleotide covalently conjugated to at least one saponin with a hydrazone linker, wherein the saponin is a bisdesomsidic triterpene with an aldehyde in position C-23 in copending claim 1, wherein the oligonucleotide is an siRNA oligonucleotide in copending claim 2, wherein the saponin is SO1861 in copending claim 7, wherein the bond is a hydrazone or hydrazine bond subject to cleavage under acidic conditions in copending claim 14, wherein the cleavage under acidic conditions occurs in vivo in the endosomes and/or lysosomes in copending claim 17, wherein the saponin is covalently bound to the aldehyde in position C-23 in copending claim 19, wherein one or more saponins are conjugated in copending claim 24, wherein the conjugate further comprises an antibody that binds EGFR or CD71 in claims 26 and 28.
‘476 copending claims 1-2, 5, 8, 10-11, 16-19, 23-26, 29-30, and 32 claim a scaffold covalently bound to an endosomal escape enhancer saponin of a 12, 13-dehydrooleanane with an aldehyde at C23 covalently coupled to the scaffold via a hydrazone bond coupled to a proteinaceous carrier molecule, wherein the scaffold is covalently attached to an immunoglobin, an effector molecule of siRNA, and wherein the biologically active molecule is a saponin.
‘476 taught a scaffold covalently bound to an endosomal escape enhancer saponin of a 12, 13-dehydrooleanane with an aldehyde at C23 covalently coupled to the scaffold via a hydrazone bond coupled to a proteinaceous carrier molecule in copending claim 1, wherein the biologically active molecule is SO1861 in copending claim 5, wherein the covalent bond is a hydrazone or hydrazide cleavable bond subject to cleavage in vivo under acidic conditions in copending claim 8, wherein the aldehyde bond in position C-23 of the saponin is involved in the covalent bond to the scaffold or the C3beta-OH group of the saponin is involved in the covalent bond to the scaffold in copending claim 11, wherein the at least one biologically active molecule has 1-128 glycoside molecules in copending claim 17, wherein the carrier molecule is a VHH domain and an anti-CD71 antibody or anti-EGFR antibody in copending claim 19, wherein the carrier molecule is covalently bound to the scaffold via a cysteine in copending claim 23, wherein the carrier molecule comprises an effector molecule of at least one oligonucleotide in copending claim 24, wherein the oligonucleotide comprises siRNA in copending claim 26,
‘759 copending claims 1, 5-6, 9, 12-13, 20-24, 29-31, 35, 38, and 43 claimed a therapeutic molecule comprising a covalent linked saponin and a covalent linked effector moiety.
‘759 taught a therapeutic molecule comprising a saponin linked to an effector moiety, wherein the saponin is a 12,13-dehydrooleanane bisdesmosidic triterpene saponin with an aldehyde function in position C-23 and optionally comprising a glucuronic acid function in a carbohydrate substituent at the C-3beta-OH group of the saponin, wherein the effector A1 comprises an oligonucleotide and the ligand B1 comprises a VHH in copending claim 1, wherein the ligand binds EGFR or CD71 in copending claim 5, wherein the ligand comprises cetuximab in copending claim 6, wherein the effector molecule is an siRNA in copending claim 9, wherein the saponin is an SO1861 in copending claim 13, wherein more than one saponin is covalently bound in copending claim 20, wherein the saponin C is a bisdesmosidic triterpene saponin with an aldehyde function in position C-23, wherein the saponin C is covalently coupled to an amino-acid residue of the first ligand at said aldehyde function in position C-23, via a cleavable linker comprising a hydrazone bond or a hydrazide bond subject to cleavage under acidic conditions when bound to saponin in copending claims 22-23, in a pharmaceutical composition in copending claim 35;
‘019 copending claims 1-2, 5, 9, 11, 17, 19, 23-24, 26, 28-29, 39-43 claims a conjugate comprising a cell-surface molecule targeting molecule and at least one effector moiety and further comprising at least one covalently bound saponin, wherein the at least one 12,13-dehydrooleanane bisdesmosidic triterpene saponin with an aldehyde function in position C-23 and comprising a glucuronic acid function in a carbohydrate substituent at the C-3beta-OH group of the saponin, and wherein the cell-surface molecule targeting molecule comprises a VHH domain, and wherein the at least one effector moiety comprises an siRNA.
‘019 taught a conjugate comprising a cell-surface molecule targeting molecule and at least one effector moiety and further comprising at least one covalently bound saponin, wherein the at least one saponin is a bisdesmosidic triterpene saponin belonging to the type of a 12,13-dehydrooleanane with an aldehyde function in position C-23 and comprising a glucuronic acid function in a carbohydrate substituent at the C-3beta-OH group of the saponin, and wherein the cell-surface molecule targeting molecule comprises a VHH domain, and wherein the at least one effector moiety comprises an siRNA in copending claim 1, Wherein the at least one saponin is SO1861 in copending claim 5, wherein the cell-surface molecule targeting molecule can bind to CD71 in copending claim 9, wherein the cell-surface molecule targeting molecule comprises an anti-CD71 antibody in copending claim 11, wherein the at least one saponin is covalently bound to the cell-surface molecule targeting molecule, via an aldehyde function in the saponin, and/or to the at least one effector moiety, via an aldehyde function in the saponin in copending claim 17, wherein the saponin is covalently bound to an amino-acid residue of the cell-surface molecule targeting molecule and/or to the at least one effector moiety via said glucuronic acid function in copending claim 19, and wherein the linker to the saponin comprises at least one cleavable linker, wherein said cleavable linker is subject to cleavage under acidic conditions in copending claim 24, and wherein the conjugate is in a pharmaceutical composition in copending claim 43.
While the claims of ‘104, ‘476, ‘759, and ‘019 taught embodiments of a VHH targeted CD71 targeted antibody covalently conjugated an siRNA and a saponin SO1861, wherein the bond is a hydrazone or hydrazine bond subject to cleavage under acidic conditions is attached at the C-23 aldehyde, they did not teach a single embodiment of the pharmaceutical combination, but this is obvious in view of Geall, Heukers, Niewoehner, Weng, Firestone, and Marciani.
Regarding instant claims 57-59, 64-65, 67-68, 70-75, and 77-78, Geall taught an effective method of cancer treatment by silencing a protein in a cancer cell comprising administering a single pharmaceutical combination comprising:
a first conjugate comprising an EGFR targeted antibody covalently linked to an effector molecule wherein the effector molecule is an siRNA oligonucleotide; and
a second conjugate comprising an EGFR targeted antibody covalently linked to melittin (page 148, [0689]),
wherein melittin is an endosomolytic moiety that causes endosomal escape agent (page 148, [0689]), wherein the expression of KRAS in the tumor was effectively silenced from 24 to 168 hours and wherein KRAS was decreased more in tumors that were administered the combination compared to an EGFR targeted antibody covalently linked to the siRNA alone (Fig. 33). Regarding claims 57-59, 64-65, 67-68, 70-75, and 77-78, Geall taught the antibody as a single-domain antibody (sdAb) (page 1, [0019]). Regarding claims 57-59, 64-65, 67-68, 70-75, and 77-78, Geall taught the endosomolytic moiety as an endosomolytic small molecule and an endosomolytic polymer (page 73, [0389]). Geall taught targeting CD71 with an antibody siRNA conjugate (Fig. 54), wherein the CD71 targeting antibody siRNA conjugate effectively silenced HPRT expression (Fig. 86).
Heukers taught EGFR-targeted nanobody® VHH single domain conjugates are specific and potently kill cancer cells (abstract and Fig. 2). Heukers taught a biparatopic EGFR-targeted single domain antibody 7D12-9G8 that is known to be internalized (page 1442, left column, first paragraph). Heukers taught 7D12-9G8-PS induces the lysosomal trafficking (page, 1448, right column first paragraph). Heukers taught EGFR-targeted single domain antibodies showed a faster accumulation at the tumor, a more homogeneous distribution within the tumor, and a more rapid clearance of unbound molecules, compared to an anti-EGFR monoclonal antibody (page 1442, left column, first paragraph).
Niewoehner taught a bivalent transferrin receptor (TfR or CD71) antibody leads to effective lysosomal targeting, wherein the bivalent TfR antibody is the same (abstract and Fig. 2E).
Weng taught the glycoside saponin SO1861 greatly improves the efficacy of lipid based as well as non-lipid based targeted nanoplexes consisting of a targeted K16 peptide with a nucleic acid binding domain and plasmid-DNA, minicircle-DNA or small interfering RNA (siRNA) (abstract). Weng taught SO1861 augments the escape of the genetic cargo out of the intracellular compartments into the cytosol (abstract). Weng taught SO1861 induces the release of the genetic cargo out of endosomes and lysosomes (abstract), wherein the lumen of the endosomes and lysosomes are acidified (page 75, right column, last paragraph). Weng taught that SO1861 integrated into a receptor targeted nanoplex improved DNA transfection and provides improved effector properties (page 85, right column, last paragraph and Fig. 10). Weng taught that SO1861 in combination with siRNA increased siRNA silencing in cancer cells (page 85, left to right column, bridging paragraph and Fig. 9).
Weng taught an effective in vitro method for transferring an oligonucleotide effector molecule from outside a Neuro-2A cell to inside of said cell comprising:
providing a pharmaceutical composition comprising a conjugate comprising SO1861 sensitized receptor-targeted nanoplexes and an oligonucleotide effector of EGFP;
providing a Neuro-2A cells wherein the surface of the cell is targeted by the receptor-targeted nanoplexes;
contacting the cell of step b) with the pharmaceutical composition of step a),
therewith establishing the transfer of the oligonucleotide effector from outside the cell into the cell, wherein the integration of SO1861 into the nanoplex caused a tremendous shift of the median fluorescence value, and wherein the high transfection efficiency
of the SO1861 sensitized LPDS-nanoplexes is corroborated by the fluorescence
image (page 85, right column, last paragraph, Fig. 10 legend, and Fig. 10). Weng taught the RISC (RNA-Induced-Silencing Complex) is located in the cytosol (page 75-76 bridging paragraph). Weng taught to exert a silencing effect, siRNA has to escape from the endosomes/lysosomes into the cytosol because in lysosomes siRNA/DNA are degraded by nucleases (page 75-76 bridging paragraph). Weng taught: 1) If the endo-/lysosomal escape of the genetic cargo does not occur the siRNA/DNA is degraded within the lysosome thus leading to a therapeutic failure; 2) limited release of siRNA/DNA into the cytosol is one of the major obstacles for efficient siRNA/DNA delivery; and 3) development of innovative strategies to augment the endo-/lysosomal escape of siRNA/DNA and the delivery of a minimal amount of siRNA/DNA are desired for an efficient therapeutic response (page 75-76 bridging paragraph).
Firestone taught BR96-Dox is an immunoconjugate in which doxorubicin is linked via an acid-labile hydrazone to the chimeric MAb BR96 (abstract). Firestone taught BR96 bound to tumor cells, then internalized via endocytosis into lysosomes, wherein the acidic milieu of the lysozymes hydrolyzed the hydrazone linkage, releasing free Dox (abstract). Firestone taught BR96-Dox was more effective at killing tumors in vivo with less toxicity and more activity when compared to untargeted Dox (abstract).
Firestone taught conjugation of the monoclonal antibody to Dox by a hydrazone bond, wherein a hydrazone bond is reasonably stable at neutral pH, but released rapidly within lysosomes, where the pH is about 5 (page 252, right column, first paragraph).
Firestone taught BR96 DOX comprised a linker of 6-Maleimidocaproylhydrazone (page 253, left column, second paragraph).
Marciani taught conjugation of moieties to a saponin wherein the conjugation site is an aldehyde at position C23 of the aglycone core (pages 23-25, Schemes 5a-b).
Regarding instant claims 57-59, 61, 64-65, 67-68, 70-75, and 77-78, it would have been obvious for a person having ordinary skill in the art to take copending claims:
1, 2, 7, 14, 17, 28 of ‘104;
1, 5, 8, 11, 19, 23, 24, 26 of ‘476;
1, 5, 9, 13, 20, 22-23, and 35 of ‘759; and
1, 4, 5, 9, 13, 20, 22-23, 33, and 43 of ‘019,
of a CD71 targeted antibody covalently conjugated to one or more of saponin SO1861 and an siRNA, wherein the bond is a hydrazone or hydrazine bond subject to cleavage under acidic conditions, wherein the aldehyde bond in position C-23 of the saponin is involved in the covalent bond – and:
Prepare a pharmaceutical composition comprising a) an antibody covalently linked to an endosomal escape moiety; and separately b) an antibody covalently linked to an siRNA, and administer it for treatment of cancer as taught by Geall;
Use a VHH CD71 targeting moiety as taught by the copending claims, Geall, Heukers, and Niewoehner;
Exchange the attachment linker of the endosomal escape moiety for an acid sensitive hydrazone linker of Firestone for linking the payload; and
covalently conjugate the acid sensitive hydrazone linker that releases the SO1861 payload to cause endosomal escape at the C23 aldehyde of SO1861 as taught by Marciani.
This is obvious because:
Geall taught an effective method of cancer treatment by silencing a protein in a cancer cell comprising administering a single pharmaceutical combination comprising: a first conjugate comprising an EGFR targeted antibody covalently linked to an effector molecule wherein the effector molecule is an siRNA oligonucleotide; and a second conjugate comprising an EGFR targeted antibody covalently linked to melittin, wherein melittin is an endosomolytic moiety that causes endosomal escape agent, Geall taught targeting CD71, and Weng taught SO1861 is an endosomal escape agent;
i) Geall taught the antibody as targeting CD71 with an antibody siRNA conjugate effectively and as a sdAb; ii) Heukers taught a bivalent biparatopic EGFR-targeted single domain antibody 7D12-9G8 was effective and the VHH causes faster accumulation at the tumor, a more homogeneous distribution within the tumor, and a more rapid clearance of unbound molecules, compared to an anti-EGFR monoclonal antibody and is known to go through endosomal clearance and kill cancer cells; and iii) Niewoehner taught a bivalent transferrin receptor (TfR or CD71) antibody leads to effective lysosomal targeting,wherein the bivalent TfR antibody is the same. Thus, a bivalent CD71 sdAb would be obvious and expected to be effective;
Firestone taught BR96 bound to tumor cells, then internalized via endocytosis into lysosomes, wherein the acidic milieu of the lysozymes hydrolyzed the hydrazone linkage, releasing free payload effectively into the cancer cells. Firestone taught conjugation of the monoclonal antibody to Dox by a hydrazone bond, wherein a hydrazone bond is stable at neutral pH, but released rapidly within lysosomes. Further, Weng taught SO1861 induces the release of the genetic cargo out of endosomes and lysosomes, wherein the lumen of the endosomes and lysosomes are acidified. Thus, the hydrazone linker would release the SO1861 payload in the endosomes/lysosomes, where SO1861 has its effects; and
Marciani taught conjugation of moieties to a saponin wherein the conjugation site is an aldehyde at position C23 of the aglycone core.
There is a reasonable expectation of success because:
A pharmaceutical composition comprising an endosomal escape moiety and an siRNA on a targeting agent separately was effective wherein the expression of KRAS in the tumor was effectively silenced from 24 to 168 hours and wherein KRAS was decreased more in tumors that were administered the combination compared to a targeted antibody covalently linked to the siRNA alone, and EGFR and CD71 targeted antibodies were both effective;
i) Geall taught the antibody as targeting CD71 with an antibody siRNA conjugate effectively and as a sdAb; ii) Heukers taught a bivalent biparatopic EGFR-targeted single domain antibody 7D12-9G8 was effective and the VHH causes faster accumulation at the tumor, a more homogeneous distribution within the tumor, and a more rapid clearance of unbound molecules, compared to an anti-EGFR monoclonal antibody and is known to go through endosomal clearance and kill cancer cells; and iii) Niewoehner taught a bivalent transferrin receptor (TfR or CD71) antibody leads to effective lysosomal targeting,wherein the bivalent TfR antibody is the same. Thus, a bivalent CD71 sdAb would be obvious and expected to be effective;
Hydrazone linkers are known to be effective, wherein hydrazone linkers are stable a neutral pH of 7, but once internalized into the acidic conditions, the payload is released into cancer cells. Further, Weng taught SO1861 induces the release of the genetic cargo out of endosomes and lysosomes, wherein the lumen of the endosomes and lysosomes are acidified. Thus, the hydrazone linker would release the SO1861 payload(s) in the endosomes/lysosomes, where SO1861 has its effects; and
an acid sensitive hydrazone linker connected at the C23 aldehyde of SO1861 would connect to the carbonyls and release the payload under acidic conditions to allow the SO1861 to perform the endosomal escape of the siRNA.
This would produce a method of cancer treatment of a subject by silencing a protein in a cancer cell comprising administering a single pharmaceutical combination (instant claim 59) comprising two pharmaceutical compositions (instant claim 58) comprising:
a first pharmaceutical composition a first conjugate comprising a VHH single domain CD71 targeted antibody, wherein the bivalent TfR VHH antibody is the same (instant claim 61), covalently linked to an effector molecule wherein the effector molecule is an siRNA oligonucleotide (instant claims 64-65); and
a second pharmaceutical composition comprising a second conjugate comprising a VHH single domain CD71 targeted antibody, wherein the bivalent TfR VHH antibody is the same, covalently linked to a linker and a modified SO1861 (instant claims 71-74) which is a bidesmosidic triterpene glycoside with an aglycone core structure of quillaic acid (instant claims 66) with a first saccharide chain of Gal-(1-2)-[Xyl-(1-3)]-GlcA- (instant claim 68), wherein the first saccharide chain would comprise a the glucuronic acid unit comprising a carboxyl group (instant claim 70), and wherein the SO1861 linker comprises an acid sensitive linker comprising a hydrazone bond linker connected at the C23 aldehyde of SO1861 (instant claims 67 and 77-78) which breaks under acidic conditions such that it forms an aldehyde function at C23 of the quillaic aglycone core structure to release the payload to cause endosomal escape (instant claims 57 and 75)
Regarding instant claim 76, it would have been obvious for a person having ordinary skill in the art to take the composition from the method of ‘104, ‘476, ‘759, or ‘019 and Geall, Heukers, Niewoehner, Weng, Firestone, and Marciani above of a pharmaceutical composition comprising two pharmaceutical compositions comprising:
a first pharmaceutical composition a first conjugate comprising a single domain CD71 targeted VHH antibody covalently linked to an effector molecule wherein the effector molecule is an siRNA oligonucleotide; and
a second pharmaceutical composition comprising a second conjugate comprising a bivalent single domain CD71 targeted VHH antibody covalently linked to a linker and a modified SO1861 which is a bidesmosidic triterpene glycoside with an aglycone core structure of quillaic acid with a first saccharide chain of Gal-(1-2)-[Xyl-(1-3)]-GlcA wherein the linker comprises an acid sensitive linker comprising a hydrazone bond linker connected at the C23 aldehyde of SO1861 which breaks under acidic conditions such that it forms an aldehyde function at C23 of the quillaic aglycone core structure to release the payload to cause endosomal escape
– and: include the compositions in the effective in vitro method for transferring an oligonucleotide effector molecule from outside an CD71 expressing cell to inside of said cell of Weng;
This is obvious because: 1) the method of Weng taught SO1861 was effective at transferring oligonucleotides into cells when targeted to the cell, Thus, the two compositions comprising SO1861 or an siRNA effector could be used for transferring nucleotides into the cell.
There is a reasonable expectation of success because: 1) SO1861 is a known endosomal escape agent that can effectively increase the effectiveness of oligonucleotide effector into the cell and the VHH would target the agents to the cells for internalization to the endosome..
This would produce an in vitro method for transferring an oligonucleotide effector molecule from outside an CD71 expressing cell to inside of said cell comprising:
providing a pharmaceutical composition comprising a bivalent single domain CD71 targeted VHH antibody covalently linked to an effector molecule wherein the effector molecule is an siRNA oligonucleotide;
providing a pharmaceutical composition comprising a second pharmaceutical composition of a second conjugate comprising a bivalent single domain CD71 targeted VHH antibody covalently linked to a linker and a modified SO1861 wherein the linker comprises an acid sensitive linker comprising a hydrazone bond linker connected at the C23 aldehyde of SO1861 which breaks under acidic conditions such that it forms an aldehyde function at C23 of the quillaic aglycone core structure to release the payload to cause endosomal escape;
providing a cell that expresses CD71 on the surface wherein the surface of the cell is targeted by the pharmaceutical compositions in a) and b);
contacting the cell of step c) in vitro with the pharmaceutical composition of step a) and the pharmaceutical composition of step b).
therewith establishing the transfer of the oligonucleotide effector from outside the cell into the cell.
This is a provisional nonstatutory double patenting rejection.
Response to Arguments
Applicant has amended the independent claim 57.
The updated to rejection is above.
Applicant requests that the provisionally rejection of the claims under the judicially created doctrine of obviousness-type double patenting as allegedly being unpatentable over the claims of co-pending applications Nos. 19/044,661, 17/312, 104, 17/312,476, 17/415,759, 17/312,019, 18/012,741, 18/012,723, 18/012,729, 18/012,760, 17/312,193 and 18/723,163 be held in abeyance until indication by the Office of allowable claims in one of the co-pending applications.
In response, Applicant's arguments filed 2/10/2026 have been fully considered
but they are not persuasive. Independent claim 57 has been amended to require a conjugate comprising a single-domain antibody is a VHH domain that can bind to CD71, wherein the first cell-surface molecule is CD71 and the updated rejection is above.
A request to hold a rejection in abeyance is not a proper response to a rejection. Rather, a request to hold a matter in abeyance may only be made in response to an OBJECTION or REQUIREMENTS AS TO FORM (see 37 CFR 1.111(b) and MPEP §714.02).
Claims 57-59, 61, 64-65, and 67-78 are provisionally rejected on the ground of nonstatutory double patenting as being unpatentable over:
claims 1-2, 5, 7, 11, 13-14, 17-19, 23-26, and 28-29 of copending Application No. 17/312,104;
claims 1-2, 5, 8, 10-11, 16-19, 23-26, 29-30, and 32 of copending Application No. 17/312,476;
claims 1, 5-6, 9, 12-13, 20-24, 29-31, 35, 38, and 43 of copending Application No. 17/415,759
claims 1-2, 5, 9, 11, 17, 19, 23-24, 26, 28-29, and 39-43 of copending Application No. 17/312,019
in view of US 2017/0281795 (Geall AJ et al. reference of record), Heukers R et al. (Nanomedicine 2014 10(7):1441-51 reference of record), Niewoehner J et al. (Neuron 2014 81(1):49-60. doi: 10.1016/j.neuron.2013.10.061.), Weng A et al. (Journal of Controlled Release 2015 206 75-90 reference of record), and Firestone RA e al. (Journal of Controlled Release 1996 39 (2-3) 251-259 reference of record), US 2004/0242502 (Marciani D et al. reference of record), and Thermo Scientific et al. Instructions BMPH, EMCH, KMUH (https://documents.thermofisher.cn/TFS-Assets%2FLSG%2Fmanuals%2FMAN0011363_BMPH_EMCH_KMUH_UG.pdf 2015 reference of record).
The claims of copending ‘104, ‘476, ‘759, or ‘019 in view of Geall, Heukers, Niewoehner, Weng, Firestone, and Marciani teach the limitations of claims 57-59, 61, 64-65, 67-68, and 70-78 for the reasons set forth above.
‘104, ‘476, ‘759, ‘019, Geall, Heukers, Niewoehner, Weng, Firestone, and Marciani are discussed above.
The claims of ‘104, ‘476, ‘759, and ‘019 did not teach conjugation of the endosomal escape moiety at a C23 of an aglycone core through reaction with EMCH, but this is obvious in view of Thermo.
Thermo taught EMCH (N-[ε-Maleimidocaproic acid) hydrazide is a heterobifunctional crosslinkers containing sulfhydryl-reactive maleimide and carbonyl-reactive hydrazide moieties, wherein hydrazide groups react with carbonyls (aldehydes and ketones) to form stable hydrazone bonds, wherein the EMCH crosslinker is useful for conjugating sulfhydryl-containing proteins (page 1, Introduction).
Regarding instant claim 69, it would have been obvious for a person having ordinary skill in the art to take the method of ‘104, ‘476, ‘759, or ‘019 in view of Geall, Heukers, Niewoehner, Weng, Firestone, and Marciani above and in the pharmaceutical composition: – 1) covalently conjugate the SO1861 payload with EMCH at an aldehyde of SO1861 as taught by Thermo, which would be the C23 aldehyde of SO1861.
This is obvious because: 1i) Thermo taught EMCH (N-[ε-Maleimidocaproic acid) hydrazide is a heterobifunctional crosslinkers containing sulfhydryl-reactive maleimide and carbonyl-reactive hydrazide moieties, wherein hydrazide groups react with carbonyls (aldehydes and ketones) to form stable hydrazone bonds, wherein the EMCH crosslinker is useful for conjugating sulfhydryl-containing proteins; and 1ii) the structure of EMCH is included in the 6-Maleimidocaproic acid hydrazone linker of Firestone.
There is a reasonable expectation of success because: 1i) Thermo taught EMCH (N-[ε-Maleimidocaproic acid) hydrazide is a heterobifunctional crosslinkers containing sulfhydryl-reactive maleimide and carbonyl-reactive hydrazide moieties, wherein hydrazide groups react with carbonyls (aldehydes and ketones) to form stable hydrazone bonds, wherein the EMCH crosslinker is useful for conjugating sulfhydryl-containing proteins; and 1ii) the structure of EMCH is included in the 6-Maleimidocaproic acid hydrazone linker of Firestone. Thus, the acid sensitive hydrazone EMCH linker connected at the C23 aldehyde of SO1861 would connect to the carbonyl and release the payload under acidic conditions to allow the SO1861 to perform the endosomal escape of the siRNA.
This is a provisional nonstatutory double patenting rejection.
Response to Arguments
Applicant has amended the independent claim 57.
The updated to rejection is above.
Applicant requests that the provisionally rejection of the claims under the judicially created doctrine of obviousness-type double patenting as allegedly being unpatentable over the claims of co-pending applications Nos. 19/044,661, 17/312, 104, 17/312,476, 17/415,759, 17/312,019, 18/012,741, 18/012,723, 18/012,729, 18/012,760, 17/312,193 and 18/723,163 be held in abeyance until indication by the Office of allowable claims in one of the co-pending applications.
In response, Applicant's arguments filed 2/10/2026 have been fully considered
but they are not persuasive. Independent claim 57 has been amended to require a conjugate comprising a single-domain antibody is a VHH domain that can bind to CD71, wherein the first cell-surface molecule is CD71 and the updated rejection is above.
A request to hold a rejection in abeyance is not a proper response to a rejection. Rather, a request to hold a matter in abeyance may only be made in response to an OBJECTION or REQUIREMENTS AS TO FORM (see 37 CFR 1.111(b) and MPEP §714.02).
Claims 57-59, 61, 64-65, 67-68, and 70-78 are provisionally rejected on the ground of nonstatutory double patenting as being unpatentable over:
claims 57-65 and 67-78 of copending Application No. 18/012,741;
claims 57-77 of copending Application No. 18/012,723; or
claims 57-80 of copending Application No. 18/012,729.
in view of US 2017/0281795 (Geall AJ et al. reference of record), Heukers R et al. (Nanomedicine 2014 10(7):1441-51 reference of record), Niewoehner J et al. (Neuron 2014 81(1):49-60. doi: 10.1016/j.neuron.2013.10.061.), Weng A et al. (Journal of Controlled Release 2015 206 75-90 reference of record), Firestone RA e al. (Journal of Controlled Release 1996 39 (2-3) 251-259 reference of record), and US 2004/0242502 (Marciani D et al. reference of record).
‘741 copending claims 57-65 and 67-78 claim a pharmaceutical combination comprising: - a first conjugate comprising an effector molecule and a single-domain antibody for binding to a first cell-surface molecule (sdAb), wherein the effector molecule and the sdAb are covalently linked to each other and wherein the effector molecule comprises an oligonucleotide;
- a second conjugate comprising a quillaic acid or gypsogenin saponin with an aldehyde or an acid sensitive bond at C23, wherein the saponin is a monodesmosidic triterpene glycoside or a bidesmosidic triterpene glycoside, wherein the saponin and the binding molecule are covalently linked together;
‘741 taught a pharmaceutical combination comprising: - a first conjugate comprising an effector molecule and a single-domain antibody for binding to a first cell-surface molecule (sdAb), wherein the effector molecule and the sdAb are covalently linked to each other and wherein the effector molecule comprises an oligonucleotide; - a second conjugate comprising a quillaic acid or gypsogenin saponin with an aldehyde or an acid sensitive bond at C23, wherein the saponin is a monodesmosidic triterpene glycoside or a bidesmosidic triterpene glycoside, wherein the saponin and the binding molecule are covalently linked together in copending claim 57, wherein the pharmaceutical combination comprises two pharmaceutical compositions in copending claim 58, wherein the first conjugate and second conjugate are provided in a single pharmaceutical composition in copending claim 59, wherein the first conjugate is a VHH domain in copending claim 60, wherein two sdAbs are the same or are capable of binding to the same binding site on the first cell-surface molecule in copending claim 61, wherein the cell surface receptor is CD71 in copending claim 62, wherein the first conjugate comprises an sdAb that can bind to CD71 in copending claim 63, wherein the effector molecule is an siRNA in copending claims 64-65, wherein the saponin is SO1861 in copending claims 68-69, wherein the saponin is covalently linked to the binding molecule via a linker that is an acid sensitive linker in copending claims 72-73, wherein the acid-sensitive linker is covalently bound to the saponin at position C23 of the aglycone core structure via an acid- sensitive bond in copending claim 74;
‘741 taught a method of treating a disease of cancer in a patient in need thereof comprising administering to the patient an effective amount of the pharmaceutical combination of claim 57 in copending claim 75.
‘741 taught an in vitro method for transferring an oligonucleotide effector molecule being an from outside a cell to inside said cell comprising: a) providing the first conjugate of the pharmaceutical combination above; b) providing the second conjugate of the pharmaceutical combination above; c) providing a cell which expresses (i) the first cell-surface molecule as recognized by the sdAb of the first conjugate and (ii) the second cell-surface molecule as recognized by the binding molecule of the second conjugate; d) contacting the cell of step c) in vitro with the first conjugate of step a) and the second conjugate of step b), therewith establishing the transfer of the effector molecule from outside the cell into said cell in copending claim 78.
‘723 copending claims 57-76 claim a conjugate for transferring an effector molecule from outside a cell into said cell, the conjugate comprising: - an effector molecule comprising or consisting of an oligonucleotide; - at least one single-domain antibody capable of binding to a binding site on a cell- surface molecule of said cell (sdAb), and - at least one saponin, wherein: the effector molecule, the sdAb, and the saponin are covalently bound to each other, wherein the covalent binding of the saponin is made via a linker, and the saponin is a mono-desmosidic triterpene glycoside or is a bi-desmosidic triterpene glycoside.
‘723 taught a conjugate for transferring an effector molecule from outside a cell into said cell, the conjugate comprising: - an effector molecule comprising or consisting of an oligonucleotide; - at least one single-domain antibody capable of binding to a binding site on a cell- surface molecule of said cell (sdAb), and - at least one saponin, wherein: the effector molecule, the sdAb, and the saponin are covalently bound to each other, wherein the covalent binding of the saponin is made via a linker, and the saponin is a mono-desmosidic triterpene glycoside or is a bi-desmosidic triterpene glycoside in copending claim 5, wherein the sdAb is a VHH domain in copending claim 58, wherein the conjugate comprises at least two sdAbs, wherein two sdAbs are the same in copending claim 59, wherein the cell-surface molecule is a cell-surface receptor and/or a tumor-cell specific receptor in copending claim 60, wherein the sdAb anti-EGFR sdAb in copending claim 61, wherein the sdAb is an anti-CD71 sdAb in copending claim 62, wherein the oligonucleotide is an siRNA in copending claim 63, wherein gene silencing targets are further claimed in copending claim 64, wherein the saponin comprises a quillaic acid aglycone core structure wherein C23 of the aglycone core structure is an aldehyde or an acid-sensitive bond that breaks under acidic conditions such that it forms an aldehyde at C23 of the aglycone core structure in copending claim 66, wherein the saponin is SO1861 in copending claims 68-69, wherein the effector molecule is covalently bound to the sdAb and/or to the saponin in copending claim 70, wherein the linker covalently binding the saponin is an acid-sensitive linker in copending claim 71, wherein the acid-sensitive linker is covalently bound to the saponin at position C23 of the aglycone core structure via an acid-sensitive bond in copending claim 71, wherein the conjugate further comprises an oligomeric or polymeric molecule to which more than one saponin is bound in copending claim 74.
‘723 taught a method for treating a disease of cancer in a patient in need thereof comprising administering to the patient an effective amount of the conjugate comprising - an effector molecule comprising or consisting of an oligonucleotide; - at least one single-domain antibody capable of binding to a binding site on a cell- surface molecule of said cell (sdAb), and - at least one saponin, wherein: the effector molecule, the sdAb, and the saponin are covalently bound to each other, wherein the covalent binding of the saponin is made via a linker, and the saponin is a mono-desmosidic triterpene glycoside or is a bi-desmosidic triterpene glycoside in copending claim 76.
‘723 taught an in vitro method for transferring an oligonucleotide from outside a cell to inside said cell comprising:
providing the conjugate of
- an effector molecule comprising or consisting of an oligonucleotide; - at least one single-domain antibody capable of binding to a binding site on a cell- surface molecule of said cell (sdAb), and - at least one saponin, wherein: the effector molecule, the sdAb, and the saponin are covalently bound to each other, wherein the covalent binding of the saponin is made via a linker, and the saponin is a mono-desmosidic triterpene glycoside or is a bi-desmosidic triterpene glycoside;
- providing a cell which expresses on its surface the binding site for the sdAb of the conjugate; and contacting the cell with the conjugate in copending claim 77
‘729 copending claims 57-80 claimed a conjugate for transferring an effector molecule from outside a cell into said cell, the conjugate comprising: a) an effector molecule that is a pharmaceutically active substance; b) at least one single-domain antibody capable of binding to a binding site on a cell-surface molecule of said cell; and c) at least one saponin, wherein the effector molecule, the sdAb, and the saponin are covalently bound to each other, wherein the covalent binding of the saponin is made via a linker, and the saponin is a mono-desmosidic triterpene glycoside or is a bi-desmosidic triterpene glycoside.
‘729 taught a conjugate for transferring an effector molecule from outside a cell into said cell, the conjugate comprising: a) an effector molecule that is a pharmaceutically active substance; b) at least one single-domain antibody capable of binding to a binding site on a cell-surface molecule of said cell; and c) at least one saponin, wherein the effector molecule, the sdAb, and the saponin are covalently bound to each other, wherein the covalent binding of the saponin is made via a linker, and the saponin is a mono-desmosidic triterpene glycoside or is a bi-desmosidic triterpene glycoside in copending claim 57, wherein the sdAb is a VHH domain in copending claim 58, wherein the conjugate comprises at least two sdAb that are the same in copending claim 59, wherein the cell surface molecule is a cell surface receptor in copending claim 60, wherein the sdAb is an antiCD71 antibody or an ant-EGFR antibody in copending claim 61, wherein the oligonucleotide is a nucleic acid in copending claim 63, wherein the oligonucleotide is a siRNA in copending claim 65, wherein the saponin is SO1861 in copending claims 72-73, wherein the effector molecule is covalently bound to the sdAb and to the saponin in copending claim 74, wherein the linker covalently bound to the saponin is an acid sensitive linker in copending claim 75, wherein the saponin comprises the first saccharide chain bound at the C3beta-OH group of the aglycone core structure and the first saccharide chain comprises a glucuronic acid moiety, wherein the saponin is covalently bound to the sdAb and/or the effector molecule via carboxyl group of the glucuronic acid moiety in the first saccharide chain, optionally via a linker covalently bound to the saponin that is an acid sensitive linker in copending claim 76,
‘729 taught a method of treating the disease of cancer by administering a conjugate for transferring an effector molecule from outside a cell into said cell, the conjugate comprising: a) an effector molecule that is a pharmaceutically active substance; b) at least one single-domain antibody capable of binding to a binding site on a cell-surface molecule of said cell; and c) at least one saponin, wherein the effector molecule, the sdAb, and the saponin are covalently bound to each other, wherein the covalent binding of the saponin is made via a linker, and the saponin is a mono-desmosidic triterpene glycoside or is a bi-desmosidic triterpene glycoside in copending claim 76.
‘729 taught an in vitro method for transferring an oligonucleotide from outside a cell to inside said cell comprising: a) providing the conjugate above; b) providing a cell which expresses on its surface the binding site for the sdAb of the conjugate; and contacting the cell with the conjugate in copending claim 80.
While the claims of ‘741, ‘723, and ‘729 taught embodiments of a CD71 targeted antibody covalently conjugated to one or more of saponin SO1861 and an siRNA, wherein the bond can undergo cleavage under acidic conditions, they did not teach a single embodiment of the pharmaceutical combination wherein the bond is a hydrazone or hydrazine bond subject to cleavage under acidic conditions is attached at the C-23 aldehyde, but this is obvious in view of Geall, Heukers, Niewoehner, Weng, Firestone, and Marciani.
Regarding instant claims 57-60, 64-65, 67-68, 70-75, and 77-78, Geall taught an effective method of cancer treatment by silencing a protein in a cancer cell comprising administering a single pharmaceutical combination comprising:
a first conjugate comprising an EGFR targeted antibody covalently linked to an effector molecule wherein the effector molecule is an siRNA oligonucleotide; and
a second conjugate comprising an EGFR targeted antibody covalently linked to melittin (page 148, [0689]),
wherein melittin is an endosomolytic moiety that causes endosomal escape agent (page 148, [0689]), wherein the expression of KRAS in the tumor was effectively silenced from 24 to 168 hours and wherein KRAS was decreased more in tumors that were administered the combination compared to an EGFR targeted antibody covalently linked to the siRNA alone (Fig. 33). Regarding claims 57-60, 64-65, 67-68, 70-75, and 77-78, Geall taught the antibody as a single-domain antibody (sdAb) (page 1, [0019]). Regarding claims 57-60, 64-65, 67-68, 70-75, and 77-78, Geall taught the endosomolytic moiety as an endosomolytic small molecule and an endosomolytic polymer (page 73, [0389]). Geall taught targeting CD71 with an antibody siRNA conjugate (Fig. 54), wherein the CD71 targeting antibody siRNA conjugate effectively silenced HPRT expression (Fig. 86).
Heukers taught EGFR-targeted nanobody® VHH single domain conjugates are specific and potently kill cancer cells (abstract and Fig. 2). Heukers taught a biparatopic EGFR-targeted single domain antibody 7D12-9G8 that is known to be internalized (page 1442, left column, first paragraph). Heukers taught 7D12-9G8-PS induces the lysosomal trafficking (page, 1448, right column first paragraph). Heukers taught EGFR-targeted single domain antibodies showed a faster accumulation at the tumor, a more homogeneous distribution within the tumor, and a more rapid clearance of unbound molecules, compared to an anti-EGFR monoclonal antibody (page 1442, left column, first paragraph).
Niewoehner taught a bivalent transferrin receptor (TfR or CD71) antibody leads to effective lysosomal targeting, wherein the bivalent TfR antibody is the same (abstract and Fig. 2E).
Weng taught the glycoside saponin SO1861 greatly improves the efficacy of lipid based as well as non-lipid based targeted nanoplexes consisting of a targeted K16 peptide with a nucleic acid binding domain and plasmid-DNA, minicircle-DNA or small interfering RNA (siRNA) (abstract). Weng taught SO1861 augments the escape of the genetic cargo out of the intracellular compartments into the cytosol (abstract). Weng taught SO1861 induces the release of the genetic cargo out of endosomes and lysosomes (abstract), wherein the lumen of the endosomes and lysosomes are acidified (page 75, right column, last paragraph). Weng taught that SO1861 integrated into a receptor targeted nanoplex improved DNA transfection and provides improved effector properties (page 85, right column, last paragraph and Fig. 10). Weng taught that SO1861 in combination with siRNA increased siRNA silencing in cancer cells (page 85, left to right column, bridging paragraph and Fig. 9).
Weng taught an effective in vitro method for transferring an oligonucleotide effector molecule from outside a Neuro-2A cell to inside of said cell comprising:
providing a pharmaceutical composition comprising a conjugate comprising SO1861 sensitized receptor-targeted nanoplexes and an oligonucleotide effector of EGFP;
providing a Neuro-2A cells wherein the surface of the cell is targeted by the receptor-targeted nanoplexes;
contacting the cell of step b) with the pharmaceutical composition of step a),
therewith establishing the transfer of the oligonucleotide effector from outside the cell into the cell, wherein the integration of SO1861 into the nanoplex caused a tremendous shift of the median fluorescence value, and wherein the high transfection efficiency
of the SO1861 sensitized LPDS-nanoplexes is corroborated by the fluorescence
image (page 85, right column, last paragraph, Fig. 10 legend, and Fig. 10). Weng taught the RISC (RNA-Induced-Silencing Complex) is located in the cytosol (page 75-76 bridging paragraph). Weng taught to exert a silencing effect, siRNA has to escape from the endosomes/lysosomes into the cytosol because in lysosomes siRNA/DNA are degraded by nucleases (page 75-76 bridging paragraph). Weng taught: 1) If the endo-/lysosomal escape of the genetic cargo does not occur the siRNA/DNA is degraded within the lysosome thus leading to a therapeutic failure; 2) limited release of siRNA/DNA into the cytosol is one of the major obstacles for efficient siRNA/DNA delivery; and 3) development of innovative strategies to augment the endo-/lysosomal escape of siRNA/DNA and the delivery of a minimal amount of siRNA/DNA are desired for an efficient therapeutic response (page 75-76 bridging paragraph).
Firestone taught BR96-Dox is an immunoconjugate in which doxorubicin is linked via an acid-labile hydrazone to the chimeric MAb BR96 (abstract). Firestone taught BR96 bound to tumor cells, then internalized via endocytosis into lysosomes, wherein the acidic milieu of the lysozymes hydrolyzed the hydrazone linkage, releasing free Dox (abstract). Firestone taught BR96-Dox was more effective at killing tumors in vivo with less toxicity and more activity when compared to untargeted Dox (abstract).
Firestone taught conjugation of the monoclonal antibody to Dox by a hydrazone bond, wherein a hydrazone bond is reasonably stable at neutral pH, but released rapidly within lysosomes, where the pH is about 5 (page 252, right column, first paragraph).
Firestone taught BR96 DOX comprised a linker of 6-Maleimidocaproylhydrazone (page 253, left column, second paragraph).
Marciani taught conjugation of moieties to a saponin wherein the conjugation site is an aldehyde at position C23 of the aglycone core (pages 23-25, Schemes 5a-b).
Regarding instant claims 57-59, 61, 64-65, 67-68, 70-75, and 77-78, it would have been obvious for a person having ordinary skill in the art to take copending claims:
57-65, 68, 72-75 of ‘741;
57-63, 66, 68-71, 74 of ‘723; and
57-63, 65, 72-76 of ‘729,
of a method of treating a disease of cancer in a patient in need thereof comprising administering to the patient an effective amount of the pharmaceutical combination comprising:
- a VHH conjugate comprising two sdAbs that are the same that target CD71 comprising an effector molecule and a saponin, wherein the effector molecule and the sdAb are covalently linked to each other and wherein the effector molecule comprises an siRNA, wherein the saponin is SO1861 conjugated via an acid sensitive linker, – and:
Prepare a pharmaceutical composition comprising a) an antibody covalently linked to an endosomal escape moiety; and separately b) an antibody covalently linked to an siRNA, and administer it for treatment of cancer as taught by Geall;
Exchange the attachment linker of the endosomal escape moiety for an acid sensitive hydrazone linker of Firestone for linking the payload;
covalently conjugate the acid sensitive hydrazone linker that releases the SO1861 payload to cause endosomal escape at the C23 aldehyde of SO1861 as taught by Marciani.
This is obvious because:
i) Geall taught an effective method of cancer treatment by silencing a protein in a cancer cell comprising administering a single pharmaceutical combination comprising: a first conjugate comprising an EGFR targeted antibody covalently linked to an effector molecule wherein the effector molecule is an siRNA oligonucleotide; and a second conjugate comprising an EGFR targeted antibody covalently linked to melittin, wherein melittin is an endosomolytic moiety that causes endosomal escape agent, Geall taught the antibody as targeting CD71 with an antibody siRNA conjugate effectively and as a sdAb; ii) Heukers taught a bivalent biparatopic EGFR-targeted single domain antibody 7D12-9G8 was effective and the VHH causes faster accumulation at the tumor, a more homogeneous distribution within the tumor, and a more rapid clearance of unbound molecules, compared to an anti-EGFR monoclonal antibody and is known to go through endosomal clearance and kill cancer cells; iii) Niewoehner taught a bivalent transferrin receptor (TfR or CD71) antibody leads to effective lysosomal targeting,wherein the bivalent TfR antibody is the same; and iv) Weng taught SO1861 is an endosomal escape agent;
Firestone taught BR96 bound to tumor cells, then internalized via endocytosis into lysosomes, wherein the acidic milieu of the lysozymes hydrolyzed the hydrazone linkage, releasing free payload effectively into the cancer cells. Firestone taught conjugation of the monoclonal antibody to Dox by a hydrazone bond, wherein a hydrazone bond is stable at neutral pH, but released rapidly within lysosomes. Further, Weng taught SO1861 induces the release of the genetic cargo out of endosomes and lysosomes, wherein the lumen of the endosomes and lysosomes are acidified. Thus, the hydrazone linker would release the SO1861 payload in the endosomes/lysosomes, where SO1861 has its effects; and
Marciani taught conjugation of moieties to a saponin wherein the conjugation site is an aldehyde at position C23 of the aglycone core.
There is a reasonable expectation of success because:
A pharmaceutical composition comprising an endosomal escape moiety and an siRNA on a targeting agent separately was effective wherein the expression of KRAS in the tumor was effectively silenced from 24 to 168 hours and wherein KRAS was decreased more in tumors that were administered the combination compared to a targeted antibody covalently linked to the siRNA alone, and CD71 targeted antibodies were effective;
Hydrazone linkers are known to be effective, wherein hydrazone linkers are stable a neutral pH of 7, but once internalized into the acidic conditions, the payload is released into cancer cells. Further, Weng taught SO1861 induces the release of the genetic cargo out of endosomes and lysosomes, wherein the lumen of the endosomes and lysosomes are acidified. Thus, the hydrazone linker would release the SO1861 payload(s) in the endosomes/lysosomes, where SO1861 has its effects; and
an acid sensitive hydrazone linker connected at the C23 aldehyde of SO1861 would connect to the carbonyls and release the payload under acidic conditions to allow the SO1861 to perform the endosomal escape of the siRNA.
This would produce a method of cancer treatment of a subject by silencing a protein in a cancer cell comprising administering a single pharmaceutical combination (instant claim 59) comprising two pharmaceutical compositions (instant claim 58) comprising:
a first pharmaceutical composition a first conjugate comprising a VHH single domain CD71 targeted antibody, wherein the bivalent TfR VHH antibody is the same (instant claim 61), covalently linked to an effector molecule wherein the effector molecule is an siRNA oligonucleotide (instant claims 64-65); and
a second pharmaceutical composition comprising a second conjugate comprising a VHH single domain CD71 targeted antibody, wherein the bivalent TfR VHH antibody is the same, covalently linked to a linker and a modified SO1861 (instant claims 71-74) which is a bidesmosidic triterpene glycoside with an aglycone core structure of quillaic acid (instant claims 66) with a first saccharide chain of Gal-(1-2)-[Xyl-(1-3)]-GlcA- (instant claim 68), wherein the first saccharide chain would comprise a the glucuronic acid unit comprising a carboxyl group (instant claim 70), and wherein the SO1861 linker comprises an acid sensitive linker comprising a hydrazone bond linker connected at the C23 aldehyde of SO1861 (instant claims 67 and 77-78) which breaks under acidic conditions such that it forms an aldehyde function at C23 of the quillaic aglycone core structure to release the payload to cause endosomal escape (instant claims 57 and 75).
Regarding instant claim 76, it would have been obvious for a person having ordinary skill in the art to take the composition from the method of ‘741, ‘723, or ‘729 and Geall, Heukers, Niewoehner, Weng, Firestone, and Marciani above of a pharmaceutical composition comprising two pharmaceutical compositions comprising:
a first pharmaceutical composition a first conjugate comprising a bivalent single domain CD71 targeted VHH antibody covalently linked to an effector molecule wherein the effector molecule is an siRNA oligonucleotide; and
a second pharmaceutical composition comprising a second conjugate comprising a bivalent single domain CD71 targeted VHH antibody covalently linked to a linker and a modified SO1861 which is a bidesmosidic triterpene glycoside with an aglycone core structure of quillaic acid with a first saccharide chain of Gal-(1-2)-[Xyl-(1-3)]-GlcA wherein the linker comprises an acid sensitive linker comprising a hydrazone bond linker connected at the C23 aldehyde of SO1861 which breaks under acidic conditions such that it forms an aldehyde function at C23 of the quillaic aglycone core structure to release the payload to cause endosomal escape
– and: 1) use it in the in vitro method taught by: A) ‘741 copending claim 78; B) ‘723 copending claim 76; or C) ‘729 copending claim 80, comprising:
an in vitro method for transferring an oligonucleotide from outside a cell to inside said cell comprising:
a) providing the conjugate above;
b) providing a cell which expresses on its surface the binding site for the sdAb of the conjugate; and contacting the cell with the conjugate; and
2) include the compositions in the effective in vitro method for transferring an oligonucleotide effector molecule from outside an CD71 expressing cell to inside of said cell of Weng;
This is obvious because the copending applications taught an in vitro method and: 1) the method of Weng taught SO1861 was effective at transferring oligonucleotides into cells when targeted to the cell, Thus, the two compositions comprising SO1861 or an siRNA effector could be used for transferring nucleotides into the cell.
There is a reasonable expectation of success because: 1) SO1861 is a known endosomal escape agent that can effectively increase the effectiveness of oligonucleotide effector into the cell and the VHH would target the agents to the cells for internalization to the endosome.
This is a provisional nonstatutory double patenting rejection.
Response to Arguments
Applicant has amended the independent claim 57.
The updated to rejection is above.
Applicant requests that the provisionally rejection of the claims under the judicially created doctrine of obviousness-type double patenting as allegedly being unpatentable over the claims of co-pending applications Nos. 19/044,661, 17/312, 104, 17/312,476, 17/415,759, 17/312,019, 18/012,741, 18/012,723, 18/012,729, 18/012,760, 17/312,193 and 18/723,163 be held in abeyance until indication by the Office of allowable claims in one of the co-pending applications.
In response, Applicant's arguments filed 2/10/2026 have been fully considered
but they are not persuasive. Independent claim 57 has been amended to require a conjugate comprising a single-domain antibody is a VHH domain that can bind to CD71, wherein the first cell-surface molecule is CD71 and the updated rejection is above.
A request to hold a rejection in abeyance is not a proper response to a rejection. Rather, a request to hold a matter in abeyance may only be made in response to an OBJECTION or REQUIREMENTS AS TO FORM (see 37 CFR 1.111(b) and MPEP §714.02).
Claims 57-59, 61, 64-65, and 67-78 are provisionally rejected on the ground of nonstatutory double patenting as being unpatentable over:
claims 57-65 and 67-78 of copending Application No. 18/012,741;
claims 57-77 of copending Application No. 18/012,723; or
claims 57-80 of copending Application No. 18/012,729.
in view of US 2017/0281795 (Geall AJ et al. reference of record), Heukers R et al. (Nanomedicine 2014 10(7):1441-51 reference of record), Niewoehner J et al. (Neuron 2014 81(1):49-60. doi: 10.1016/j.neuron.2013.10.061.), Weng A et al. (Journal of Controlled Release 2015 206 75-90 reference of record), Firestone RA e al. (Journal of Controlled Release 1996 39 (2-3) 251-259 reference of record), US 2004/0242502 (Marciani D et al. reference of record), and Thermo Scientific et al. Instructions BMPH, EMCH, KMUH (https://documents.thermofisher.cn/TFS-Assets%2FLSG%2Fmanuals%2FMAN0011363_BMPH_EMCH_KMUH_UG.pdf 2015 reference of record).
The claims of copending ‘741, ‘723, or ‘729 in view of Geall, Heukers, Niewoehner, Weng, Firestone, and Marciani teach the limitations of claims 57-59, 61, 64-65, and 67-68, and 70-78 for the reasons set forth above.
‘741, ‘723, or ‘729, Geall, Heukers, Niewoehner, Weng, Firestone, and Marciani are discussed above.
The claims of ‘741, ‘723, or ‘729 did not teach conjugation of the endosomal escape moiety at a C23 of an aglycone core through reaction with EMCH, but this is obvious in view of Thermo.
Thermo taught EMCH (N-[ε-Maleimidocaproic acid) hydrazide is a heterobifunctional crosslinkers containing sulfhydryl-reactive maleimide and carbonyl-reactive hydrazide moieties, wherein hydrazide groups react with carbonyls (aldehydes and ketones) to form stable hydrazone bonds, wherein the EMCH crosslinker is useful for conjugating sulfhydryl-containing proteins (page 1, Introduction).
Regarding instant claim 69, it would have been obvious for a person having ordinary skill in the art to take the method of ‘741, ‘723, or ‘729, and Geall, Heukers, Niewoehner, Weng, Firestone, and Marciani above and in the pharmaceutical composition: – 1) covalently conjugate the SO1861 payload with EMCH at an aldehyde of SO1861 as taught by Thermo, which would be the C23 aldehyde of SO1861.
This is obvious because: 1i) Thermo taught EMCH (N-[ε-Maleimidocaproic acid) hydrazide is a heterobifunctional crosslinkers containing sulfhydryl-reactive maleimide and carbonyl-reactive hydrazide moieties, wherein hydrazide groups react with carbonyls (aldehydes and ketones) to form stable hydrazone bonds, wherein the EMCH crosslinker is useful for conjugating sulfhydryl-containing proteins; and 1ii) the structure of EMCH is included in the 6-Maleimidocaproic acid hydrazone linker of Firestone.
There is a reasonable expectation of success because: 1i) Thermo taught EMCH (N-[ε-Maleimidocaproic acid) hydrazide is a heterobifunctional crosslinkers containing sulfhydryl-reactive maleimide and carbonyl-reactive hydrazide moieties, wherein hydrazide groups react with carbonyls (aldehydes and ketones) to form stable hydrazone bonds, wherein the EMCH crosslinker is useful for conjugating sulfhydryl-containing proteins; and 1ii) the structure of EMCH is included in the 6-Maleimidocaproic acid hydrazone linker of Firestone. Thus, the acid sensitive hydrazone EMCH linker connected at the C23 aldehyde of SO1861 would connect to the carbonyl and release the payload under acidic conditions to allow the SO1861 to perform the endosomal escape of the siRNA.
This is a provisional nonstatutory double patenting rejection.
Response to Arguments
Applicant has amended the independent claim 57.
The updated to rejection is above.
Applicant requests that the provisionally rejection of the claims under the judicially created doctrine of obviousness-type double patenting as allegedly being unpatentable over the claims of co-pending applications Nos. 19/044,661, 17/312, 104, 17/312,476, 17/415,759, 17/312,019, 18/012,741, 18/012,723, 18/012,729, 18/012,760, 17/312,193 and 18/723,163 be held in abeyance until indication by the Office of allowable claims in one of the co-pending applications.
In response, Applicant's arguments filed 2/10/2026 have been fully considered
but they are not persuasive. Independent claim 57 has been amended to require a conjugate comprising a single-domain antibody is a VHH domain that can bind to CD71, wherein the first cell-surface molecule is CD71 and the updated rejection is above.
A request to hold a rejection in abeyance is not a proper response to a rejection. Rather, a request to hold a matter in abeyance may only be made in response to an OBJECTION or REQUIREMENTS AS TO FORM (see 37 CFR 1.111(b) and MPEP §714.02).
Claims 57-59, 61, 64-65, and 67-68, 70-78 are provisionally rejected on the ground of nonstatutory double patenting as being unpatentable over:
claims 1-6, 8-9, 11-14, 17, 21-26, and 54 of copending Application No. 18/012,760;
claims 1, 4, 7-11, 13-14, 16, 19, 26-30, 32-34, 43, 49-50 of copending Application No. 17/312,193; or
claims 29-31, 37, 39, 41, 45-50, 52, and 58-64 of copending Application No. 18/723,163;
in view of US 2017/0281795 (Geall AJ et al. reference of record), Heukers R et al. (Nanomedicine 2014 10(7):1441-51 reference of record), Niewoehner J et al. (Neuron 2014 81(1):49-60. doi: 10.1016/j.neuron.2013.10.061.), Weng A et al. (Journal of Controlled Release 2015 206 75-90 reference of record), Firestone RA e al. (Journal of Controlled Release 1996 39 (2-3) 251-259 reference of record), and US 2004/0242502 (Marciani D et al. reference of record).
Copending ‘760 claims 1-6, 8-9, 11-14, 17, 21-26, and 54 claimed a therapeutic or pharmaceutical combination comprising: (a) a first pharmaceutical composition comprising a conjugate comprising a first binding molecule comprising a first binding region for binding to a first binding site of a cell- surface molecule and the conjugate comprising at least one saponin covalently bound to said first binding molecule, wherein the saponin is a monodesmosidic triterpene glycoside or a bidesmosidic triterpene glycoside; and (b) a second pharmaceutical composition comprising a conjugate comprising a second binding molecule different from the first binding molecule, the second binding molecule comprising a second binding region different from the first binding region, the second binding region for binding to a second binding site of said cell-surface molecule different from the first binding site of said cell-surface molecule, and the conjugate comprising an effector molecule covalently bound to said second binding molecule,
‘760 taught a therapeutic or pharmaceutical combination comprising: (a) a first pharmaceutical composition comprising a conjugate comprising a first binding molecule comprising a first binding region for binding to a first binding site of a cell- surface molecule and the conjugate comprising at least one saponin covalently bound to said first binding molecule, wherein the saponin is a monodesmosidic triterpene glycoside or a bidesmosidic triterpene glycoside; and (b) a second pharmaceutical composition comprising a conjugate comprising a second binding molecule different from the first binding molecule, the second binding molecule comprising a second binding region different from the first binding region, the second binding region for binding to a second binding site of said cell-surface molecule different from the first binding site of said cell-surface molecule, and the conjugate comprising an effector molecule covalently bound to said second binding molecule in copending claims 1-2, wherein the first and second binding molecules are proteinaceous binding molecules in copending claim 3, wherein the saponin of the first binding molecule is covalently bound in copending claim 4, wherein the first binding site is a first epitope of said cell surface molecule and the second binding site is a second epitope of the same cell surface molecule that is a different epitope from the first epitope in copending claim 5, wherein the saponin is a bisdesomsidic triterpene saponin in copending claim 6, wherein the first and second binding sites are a VHH in copending claim 9, wherein the first and second binding sites simultaneously bind the same cell surface molecule at the first and second binding site in copending claim 11, wherein the saponin is SO1861 in copending claim 17, wherein the saponin is a bisdesomsidic triterpene saponin with an aldehyde at C23 of the aglycone core that is covalently bound to the first binding molecule in copending claim 21, wherein the aldehyde is covalently bound to the linker EMCH via a thioether sulfhydryl in copending claim 22, wherein the saponin is a bisdesomsidic triterpene saponin with an aldehyde at C23 of the aglycone core with a C3beta OH saccharide glucuronic acid that is covalently bound to the first binding molecule in copending claim 23,
Copending ‘193 claims 1, 4, 7-11, 13-14, 16, 19, 26-30, 32-34, 43, and 49-50 claimed a first proteinaceous molecule comprising a first binding site for binding to a first epitope of a first cell-surface molecule, the first proteinaceous molecule provided with at least one saponin covalently bound via at least one linker or via an oligomeric or polymeric scaffold to an amino-acid residue of said first proteinaceous molecule, wherein the first binding site comprises a binding fragment of an immunoglobin.
‘193 taught a first proteinaceous molecule comprising a first binding site for binding to a first epitope of a first cell-surface molecule, the first proteinaceous molecule provided with at least one saponin covalently bound via at least one linker or via an oligomeric or polymeric scaffold to an amino-acid residue of said first proteinaceous molecule, wherein the first binding site comprises a binding fragment of an immunoglobin in copending claim 1, wherein the saponin is SO1861 in copending claim 4, wherein the saponin is a bidesmosidic triterpene glycoside with an aldehyde function in position C-23 covalently coupling the saponin to the proteinaceous molecule in copending claim 7, wherein the saponin is a bidesmosidic triterpene glycoside with an aldehyde function in position C-23 and a C3beta-OH glucuronic acids that covalently couples the saponin to the proteinaceous molecule in copending claim 8, wherein the C-23 linker is N-e-maleimidocaproic acid hydrazide, which is acid labile and covalently coupled to a sulfhydryl group to the fist proteinaceous molecule in copending claim 9, wherein the C3beta-OH carbohydrate is covalently coupled to a linker to which is further coupled to an amine group of a lysine to the first proteinaceous molecule in copending claim 10, wherein the first epitope of the first cell surface molecule which binds the first proteinaceous molecule is CD71 in copending claim 11, wherein the first proteinaceous molecule is cetuximab in copending claims 13 and 26, wherein the saponin comprises more than one saponin covalently bound to a linker in copending claim 32, wherein the linker is a linker that is subject to cleavage under acidic conditions in copending claims 33-34, and wherein the proteinaceous molecule binds CD71 in copending 43.
‘163 claims 29-31, 37, 39, 41, 45-50, 52, 58-64 claimed a therapeutic combination comprising: a) a nucleic acid; and b) a saponin, wherein the saponin is a triterpenoid 12,13 -dehydrooleanane-type saponin comprising an aldehyde group at position C-23 of the saponin' s aglycone core structure under acidic conditions present in endosomes and/or lysosomes of human cells.
‘163 taught a therapeutic combination comprising: a) a nucleic acid; and b) a saponin, wherein the saponin is a triterpenoid 12,13 -dehydrooleanane-type saponin comprising an aldehyde group at position C-23 of the saponin' s aglycone core structure under acidic conditions present in endosomes and/or lysosomes of human cells in copending claim 29, wherein the aldehyde group at position C-23 of the saponin' s aglycone core structure is either a free aldehyde group, or is an aldehyde group substituted by a maleimide-comprising moiety attached at said position C-23 with a cleavable covalent bond that cleaves off under acidic conditions present in endosomes and/or lysosomes of human cells; and, wherein said aldehyde group at position C-23 of the saponin's aglycone core structure is restored upon said cleavage under acidic conditions present in endosomes and/or lysosomes of human cells in copending claim 30, wherein the maleimide-comprising moiety is a part of a molecule comprising 4- (6-(2,5-dioxo-2,5-dihydro- 1H-pyrrol- 1-yl)hexanoyl)piperazine- 1-carbohydrazide that is attached at position C-23 of the saponin's aglycone core structure upon forming a semicarbazone bond (SC-Maleimide); or wherein the maleimide-comprising moiety is a part of a molecule comprising N-e-maleimidocaproic acid (EMCH) hydrazide that is attached at position C-23 of the saponin's aglycone core structure upon forming a hydrazone bond in copending claim 31, wherein the saponin is SO1861 in copending claim 37, wherein the nucleic acid is siRNA in copending claim 41, wherein the endocytic receptor to which the ligand binds is CD71 in copending claim 45, wherein the ligand is an antibody or a binding fragment thereof specific for binding to the endocytic receptor, wherein the endocytic receptor is CD71 in copending claim 60, and wherein the ligand is a single domain antibody specific for binding to a transferrin receptor in copending claim 62.
‘760, ‘193, or ‘163 did not teach a pharmaceutical composition of a first conjugate comprising a sdAb-siRNA effector and a second conjugate covalently linked to a saponin at the C23 aldehyde, but this is obvious in view of Geall, Heukers, Niewoehner, Weng, Firestone, and Marciani.
Regarding instant claims 57-60, 61, 64-65, 67-68, and 70-75, and 77-78, Geall taught an effective method of cancer treatment by silencing a protein in a cancer cell comprising administering a single pharmaceutical combination comprising:
a first conjugate comprising an EGFR targeted antibody covalently linked to an effector molecule wherein the effector molecule is an siRNA oligonucleotide; and
a second conjugate comprising an EGFR targeted antibody covalently linked to melittin (page 148, [0689]),
wherein melittin is an endosomolytic moiety that causes endosomal escape agent (page 148, [0689]), wherein the expression of KRAS in the tumor was effectively silenced from 24 to 168 hours and wherein KRAS was decreased more in tumors that were administered the combination compared to an EGFR targeted antibody covalently linked to the siRNA alone (Fig. 33). Regarding claims 57-60, 61, 64-65, 67-68, and 70-75, and 77-78, Geall taught the antibody as a single-domain antibody (sdAb) (page 1, [0019]). Regarding claims 57-60, 61, 64-65, 67-68, and 70-75, and 77-78, Geall taught the endosomolytic moiety as an endosomolytic small molecule and an endosomolytic polymer (page 73, [0389]). Geall taught targeting CD71 with an antibody siRNA conjugate (Fig. 54), wherein the CD71 targeting antibody siRNA conjugate effectively silenced HPRT expression (Fig. 86).
Heukers taught EGFR-targeted nanobody® VHH single domain conjugates are specific and potently kill cancer cells (abstract and Fig. 2). Heukers taught a biparatopic EGFR-targeted single domain antibody 7D12-9G8 that is known to be internalized (page 1442, left column, first paragraph). Heukers taught 7D12-9G8-PS induces the lysosomal trafficking (page, 1448, right column first paragraph). Heukers taught EGFR-targeted single domain antibodies showed a faster accumulation at the tumor, a more homogeneous distribution within the tumor, and a more rapid clearance of unbound molecules, compared to an anti-EGFR monoclonal antibody (page 1442, left column, first paragraph).
Niewoehner taught a bivalent transferrin receptor (TfR or CD71) antibody leads to effective lysosomal targeting, wherein the bivalent TfR antibody is the same (abstract and Fig. 2E).
Weng taught the glycoside saponin SO1861 greatly improves the efficacy of lipid based as well as non-lipid based targeted nanoplexes consisting of a targeted K16 peptide with a nucleic acid binding domain and plasmid-DNA, minicircle-DNA or small interfering RNA (siRNA) (abstract). Weng taught SO1861 augments the escape of the genetic cargo out of the intracellular compartments into the cytosol (abstract). Weng taught SO1861 induces the release of the genetic cargo out of endosomes and lysosomes (abstract), wherein the lumen of the endosomes and lysosomes are acidified (page 75, right column, last paragraph). Weng taught that SO1861 integrated into a receptor targeted nanoplex improved DNA transfection and provides improved effector properties (page 85, right column, last paragraph and Fig. 10). Weng taught that SO1861 in combination with siRNA increased siRNA silencing in cancer cells (page 85, left to right column, bridging paragraph and Fig. 9).
Weng taught an effective in vitro method for transferring an oligonucleotide effector molecule from outside a Neuro-2A cell to inside of said cell comprising:
providing a pharmaceutical composition comprising a conjugate comprising SO1861 sensitized receptor-targeted nanoplexes and an oligonucleotide effector of EGFP;
providing a Neuro-2A cells wherein the surface of the cell is targeted by the receptor-targeted nanoplexes;
contacting the cell of step b) with the pharmaceutical composition of step a),
therewith establishing the transfer of the oligonucleotide effector from outside the cell into the cell, wherein the integration of SO1861 into the nanoplex caused a tremendous shift of the median fluorescence value, and wherein the high transfection efficiency
of the SO1861 sensitized LPDS-nanoplexes is corroborated by the fluorescence
image (page 85, right column, last paragraph, Fig. 10 legend, and Fig. 10).
Weng taught the RISC (RNA-Induced-Silencing Complex) is located in the cytosol (page 75-76 bridging paragraph). Weng taught to exert a silencing effect, siRNA has to escape from the endosomes/lysosomes into the cytosol because in lysosomes siRNA/DNA are degraded by nucleases (page 75-76 bridging paragraph). Weng taught: 1) If the endo-/lysosomal escape of the genetic cargo does not occur the siRNA/DNA is degraded within the lysosome thus leading to a therapeutic failure; 2) limited release of siRNA/DNA into the cytosol is one of the major obstacles for efficient siRNA/DNA delivery; and 3) development of innovative strategies to augment the endo-/lysosomal escape of siRNA/DNA and the delivery of a minimal amount of siRNA/DNA are desired for an efficient therapeutic response (page 75-76 bridging paragraph).
Firestone taught BR96-Dox is an immunoconjugate in which doxorubicin is linked via an acid-labile hydrazone to the chimeric MAb BR96 (abstract). Firestone taught BR96 bound to tumor cells, then internalized via endocytosis into lysosomes, wherein the acidic milieu of the lysozymes hydrolyzed the hydrazone linkage, releasing free Dox (abstract). Firestone taught BR96-Dox was more effective at killing tumors in vivo with less toxicity and more activity when compared to untargeted Dox (abstract).
Firestone taught conjugation of the monoclonal antibody to Dox by a hydrazone bond, wherein a hydrazone bond is reasonably stable at neutral pH, but released rapidly within lysosomes, where the pH is about 5 (page 252, right column, first paragraph).
Firestone taught BR96 DOX comprised a linker of 6-Maleimidocaproylhydrazone (page 253, left column, second paragraph).
Marciani taught conjugation of moieties to a saponin wherein the conjugation site is an aldehyde at position C23 of the aglycone core (pages 23-25, Schemes 5a-b).
Regarding instant claims 57-60, 62, 64-68, 70-75, and 77-78, it would have been obvious for a person having ordinary skill in the art to take copending claims:
1-6, 11, 17, 21-23 of ‘760;
1, 4, 7-11, 13, 26, 32-34, and 43 of ‘193;
29-31, 37, 41, 45, 60, 62 of ‘163,
of a therapeutic combination comprising:
a) a nucleic acid of siRNA; and
b) a saponin of SO1861 attached to a binding molecule,
– and:
Prepare a pharmaceutical composition comprising a) an antibody covalently linked to an endosomal escape moiety; and separately b) an antibody covalently linked to an siRNA, and administer it for treatment of cancer as taught by Geall;
Use a VHH CD71 targeting moiety in view of the copending claims and/or Geall, Heukers, and Niewoehner; and
Exchange the attachment linker of the endosomal escape moiety for an acid sensitive hydrazone linker of Firestone for linking the payload;
covalently conjugate the acid sensitive hydrazone linker that releases the SO1861 payload to cause endosomal escape at the C23 aldehyde of SO1861 as taught by Marciani.
This is obvious because:
a) Geall taught separate siRNA and endosomal escape moieties on antibody conjugates were effective and the antibody as a sdAb and Heukers taught the bivalent biparatopic EGFR-targeted single domain antibody 7D12-9G8 was effective and the VHH causes faster accumulation at the tumor, a more homogeneous distribution within the tumor, and a more rapid clearance of unbound molecules, compared to an anti-EGFR monoclonal antibody and is known to go through endosomal clearance and kill cancer cells; b) Weng taught the saponin SO1861: i) augments the escape of the genetic cargo out of the intracellular compartments into the cytosol; ii) integrated into a receptor targeted nanoplex improved DNA transfection and provides improved effector properties; and iii) in combination with siRNA increased siRNA silencing in cancer cells;
i) Geall taught the antibody as targeting CD71 with an antibody siRNA conjugate effectively and as a sdAb; ii) Heukers taught a bivalent biparatopic EGFR-targeted single domain antibody 7D12-9G8 was effective and the VHH causes faster accumulation at the tumor, a more homogeneous distribution within the tumor, and a more rapid clearance of unbound molecules, compared to an anti-EGFR monoclonal antibody and is known to go through endosomal clearance and kill cancer cells; and iii) Niewoehner taught a bivalent transferrin receptor (TfR or CD71) antibody leads to effective lysosomal targeting,wherein the bivalent TfR antibody is the same. Thus, a bivalent CD71 sdAb would be obvious and expected to be effective;
Firestone taught BR96 bound to tumor cells, then internalized via endocytosis into lysosomes, wherein the acidic milieu of the lysozymes hydrolyzed the hydrazone linkage, releasing free payload effectively into the cancer cells. Firestone taught conjugation of the monoclonal antibody to Dox by a hydrazone bond, wherein a hydrazone bond is stable at neutral pH, but released rapidly within lysosomes. Further, Weng taught SO1861 induces the release of the genetic cargo out of endosomes and lysosomes, wherein the lumen of the endosomes and lysosomes are acidified. Thus, the hydrazone linker would release the SO1861 payload in the endosomes/lysosomes, where SO1861 has its effects; and
Marciani taught conjugation of moieties to a saponin wherein the conjugation site is an aldehyde at position C23 of the aglycone core.
There is a reasonable expectation of success because:
a) The compositions of Geall were effective and the bivalent biparatopic EGFR-targeted single domain antibody 7D12-9G8 was effective and the VHH causes faster accumulation at the tumor, a more homogeneous distribution within the tumor, and a more rapid clearance of unbound molecules, compared to an anti-EGFR monoclonal antibody and is known to go through endosomal clearance and kill cancer cells; b) The saponin SO1861 would: i) augment the escape of the siRNA genetic cargo out of the intracellular compartments into the cytosol; ii) improve DNA transfection and provide improved effector properties; and iii) increase siRNA silencing in cancer cells to promote;
i) Geall taught the antibody as targeting CD71 with an antibody siRNA conjugate effectively and as a sdAb; ii) Heukers taught a bivalent biparatopic EGFR-targeted single domain antibody 7D12-9G8 was effective and the VHH causes faster accumulation at the tumor, a more homogeneous distribution within the tumor, and a more rapid clearance of unbound molecules, compared to an anti-EGFR monoclonal antibody and is known to go through endosomal clearance and kill cancer cells; and iii) Niewoehner taught a bivalent transferrin receptor (TfR or CD71) antibody leads to effective lysosomal targeting,wherein the bivalent TfR antibody is the same. Thus, a bivalent CD71 sdAb would be obvious and expected to be effective;
Hydrazone linkers are known to be effective, wherein hydrazone linkers are stable a neutral pH of 7, but once internalized into the acidic conditions, the payload is released into cancer cells. Further, Weng taught SO1861 induces the release of the genetic cargo out of endosomes and lysosomes, wherein the lumen of the endosomes and lysosomes are acidified. Thus, the hydrazone linker would release the SO1861 payload(s) in the endosomes/lysosomes, where SO1861 has its effects; and
an acid sensitive hydrazone linker connected at the C23 aldehyde of SO1861 would connect to the carbonyls and release the payload under acidic conditions to allow the SO1861 to perform the endosomal escape of the siRNA.
This would produce a method of cancer treatment of a subject by silencing a protein in a cancer cell comprising administering a single pharmaceutical combination (instant claim 59) comprising two pharmaceutical compositions (instant claim 58) comprising:
a first pharmaceutical composition a first conjugate comprising a VHH single domain CD71 targeted antibody, wherein the bivalent TfR VHH antibody is the same (instant claim 61), covalently linked to an effector molecule wherein the effector molecule is an siRNA oligonucleotide (instant claims 64-65); and
a second pharmaceutical composition comprising a second conjugate comprising a VHH single domain CD71 targeted antibody, wherein the bivalent TfR VHH antibody is the same, covalently linked to a linker and a modified SO1861 (instant claims 71-74) which is a bidesmosidic triterpene glycoside with an aglycone core structure of quillaic acid (instant claims 66) with a first saccharide chain of Gal-(1-2)-[Xyl-(1-3)]-GlcA- (instant claim 68), wherein the first saccharide chain would comprise a the glucuronic acid unit comprising a carboxyl group (instant claim 70), and wherein the SO1861 linker comprises an acid sensitive linker comprising a hydrazone bond linker connected at the C23 aldehyde of SO1861 (instant claims 67 and 77-78) which breaks under acidic conditions such that it forms an aldehyde function at C23 of the quillaic aglycone core structure to release the payload to cause endosomal escape (instant claims 57 and 75).
Regarding instant claim 76, it would have been obvious for a person having ordinary skill in the art to take the composition from the method of ‘760, ‘193, or ‘163 and Geall, Heukers, Niewoehner, Weng, Firestone, and Marciani above of a pharmaceutical composition comprising two pharmaceutical compositions comprising:
a first pharmaceutical composition a first conjugate comprising a bivalent single domain CD71 targeted VHH antibody covalently linked to an effector molecule wherein the effector molecule is an siRNA oligonucleotide; and
a second pharmaceutical composition comprising a second conjugate comprising a bivalent single domain CD71 targeted VHH antibody covalently linked to a linker and a modified SO1861 which is a bidesmosidic triterpene glycoside with an aglycone core structure of quillaic acid with a first saccharide chain of Gal-(1-2)-[Xyl-(1-3)]-GlcA wherein the linker comprises an acid sensitive linker comprising a hydrazone bond linker connected at the C23 aldehyde of SO1861 which breaks under acidic conditions such that it forms an aldehyde function at C23 of the quillaic aglycone core structure to release the payload to cause endosomal escape
– and: include the compositions in the effective in vitro method for transferring an oligonucleotide effector molecule from outside an CD71 expressing cell to inside of said cell of Weng;
This is obvious because: 1) the method of Weng taught SO1861 was effective at transferring oligonucleotides into cells when targeted to the cell, Thus, the two compositions comprising SO1861 or an siRNA effector could be used for transferring nucleotides into the cell.
There is a reasonable expectation of success because: 1) SO1861 is a known endosomal escape agent that can effectively increase the effectiveness of oligonucleotide effector into the cell and the VHH would target the agents to the cells for internalization to the endosome..
This would produce an in vitro method for transferring an oligonucleotide effector molecule from outside an CD71 expressing cell to inside of said cell comprising:
providing a pharmaceutical composition comprising a first pharmaceutical composition a first conjugate comprising a bivalent single domain CD71 targeted VHH antibody covalently linked to an effector molecule wherein the effector molecule is an siRNA oligonucleotide;
providing a pharmaceutical composition comprising a second pharmaceutical composition of a second conjugate comprising a bivalent single domain CD71 targeted VHH antibody covalently linked to a linker and a modified SO1861 wherein the linker comprises an acid sensitive linker comprising a hydrazone bond linker connected at the C23 aldehyde of SO1861 which breaks under acidic conditions such that it forms an aldehyde function at C23 of the quillaic aglycone core structure to release the payload to cause endosomal escape;
providing a cell that expresses CD71 on the surface wherein the surface of the cell is targeted by the pharmaceutical compositions in a) and b);
contacting the cell of step c) in vitro with the pharmaceutical composition of step a) and the pharmaceutical composition of step b).
therewith establishing the transfer of the oligonucleotide effector from outside the cell into the cell.
This is a provisional nonstatutory double patenting rejection.
Response to Arguments
Applicant has amended the independent claim 57.
The updated to rejection is above.
Applicant requests that the provisionally rejection of the claims under the judicially created doctrine of obviousness-type double patenting as allegedly being unpatentable over the claims of co-pending applications Nos. 19/044,661, 17/312, 104, 17/312,476, 17/415,759, 17/312,019, 18/012,741, 18/012,723, 18/012,729, 18/012,760, 17/312,193 and 18/723,163 be held in abeyance until indication by the Office of allowable claims in one of the co-pending applications.
In response, Applicant's arguments filed 2/10/2026 have been fully considered
but they are not persuasive. Independent claim 57 has been amended to require a conjugate comprising a single-domain antibody is a VHH domain that can bind to CD71, wherein the first cell-surface molecule is CD71 and the updated rejection is above.
A request to hold a rejection in abeyance is not a proper response to a rejection. Rather, a request to hold a matter in abeyance may only be made in response to an OBJECTION or REQUIREMENTS AS TO FORM (see 37 CFR 1.111(b) and MPEP §714.02).
Claims 57-59, 61, 64-65, 67-78 are provisionally rejected on the ground of nonstatutory double patenting as being unpatentable over:
claims 1-6, 8-9, 11-14, 17, 21-26, and 54 of copending Application No. 18/012,760;
claims 1, 4, 7-11, 13-14, 16, 19, 26-30, 32-34, 43, 49-50 of copending Application No. 17/312,193; or
claims 29-31, 37, 39, 41, 45-50, 52, and 58-64 of copending Application No. 18/723,163;
in view of US 2017/0281795 (Geall AJ et al. reference of record), Heukers R et al. (Nanomedicine 2014 10(7):1441-51 reference of record), Niewoehner J et al. (Neuron 2014 81(1):49-60. doi: 10.1016/j.neuron.2013.10.061.), Weng A et al. (Journal of Controlled Release 2015 206 75-90 reference of record), Firestone RA e al. (Journal of Controlled Release 1996 39 (2-3) 251-259 reference of record), US 2004/0242502 (Marciani D et al. reference of record), and Thermo Scientific et al. Instructions BMPH, EMCH, KMUH (https://documents.thermofisher.cn/TFS-Assets%2FLSG%2Fmanuals%2FMAN0011363_BMPH_EMCH_KMUH_UG.pdf 2015 reference of record).
The claims of copending '760, '193, or '163 in view of Geall, Heukers, Niewoehner, Weng, Firestone, and Marciani teach the limitations of claims 57-59, 61, 64-65, 67-68, and 70-78 for the reasons set forth above.
'760, '193, or '163, Geall, Heukers, Niewoehner, Weng, Firestone, and Marciani are discussed above.
'760, '193, or '163 did not teach conjugation of the endosomal escape moiety at a C23 of an aglycone core through reaction with EMCH, but this is obvious in view of Thermo.
Thermo taught EMCH (N-[ε-Maleimidocaproic acid) hydrazide is a heterobifunctional crosslinkers containing sulfhydryl-reactive maleimide and carbonyl-reactive hydrazide moieties, wherein hydrazide groups react with carbonyls (aldehydes and ketones) to form stable hydrazone bonds, wherein the EMCH crosslinker is useful for conjugating sulfhydryl-containing proteins (page 1, Introduction).
Regarding instant claim 69, it would have been obvious for a person having ordinary skill in the art to take the method of ‘104, ‘476, ‘759, or ‘019 and Geall, Heukers, Niewoehner, Weng, Firestone, and Marciani above and in the pharmaceutical composition: – 1) covalently conjugate the SO1861 payload with EMCH at an aldehyde of SO1861 as taught by Thermo, which would be the C23 aldehyde of SO1861.
This is obvious because: 1i) Thermo taught EMCH (N-[ε-Maleimidocaproic acid) hydrazide is a heterobifunctional crosslinkers containing sulfhydryl-reactive maleimide and carbonyl-reactive hydrazide moieties, wherein hydrazide groups react with carbonyls (aldehydes and ketones) to form stable hydrazone bonds, wherein the EMCH crosslinker is useful for conjugating sulfhydryl-containing proteins; and 1ii) the structure of EMCH is included in the 6-Maleimidocaproic acid hydrazone linker of Firestone.
There is a reasonable expectation of success because: 1i) Thermo taught EMCH (N-[ε-Maleimidocaproic acid) hydrazide is a heterobifunctional crosslinkers containing sulfhydryl-reactive maleimide and carbonyl-reactive hydrazide moieties, wherein hydrazide groups react with carbonyls (aldehydes and ketones) to form stable hydrazone bonds, wherein the EMCH crosslinker is useful for conjugating sulfhydryl-containing proteins; and 1ii) the structure of EMCH is included in the 6-Maleimidocaproic acid hydrazone linker of Firestone. Thus, the acid sensitive hydrazone EMCH linker connected at the C23 aldehyde of SO1861 would connect to the carbonyl and release the payload under acidic conditions to allow the SO1861 to perform the endosomal escape of the siRNA.
This is a provisional nonstatutory double patenting rejection.
Response to Arguments
Applicant has amended the independent claim 57.
The updated to rejection is above.
Applicant requests that the provisionally rejection of the claims under the judicially created doctrine of obviousness-type double patenting as allegedly being unpatentable over the claims of co-pending applications Nos. 19/044,661, 17/312, 104, 17/312,476, 17/415,759, 17/312,019, 18/012,741, 18/012,723, 18/012,729, 18/012,760, 17/312,193 and 18/723,163 be held in abeyance until indication by the Office of allowable claims in one of the co-pending applications.
In response, Applicant's arguments filed 2/10/2026 have been fully considered
but they are not persuasive. Independent claim 57 has been amended to require a conjugate comprising a single-domain antibody is a VHH domain that can bind to CD71, wherein the first cell-surface molecule is CD71 and the updated rejection is above.
A request to hold a rejection in abeyance is not a proper response to a rejection. Rather, a request to hold a matter in abeyance may only be made in response to an OBJECTION or REQUIREMENTS AS TO FORM (see 37 CFR 1.111(b) and MPEP §714.02).
Conclusion
Claims 57-59, 61, 64-65, and 67-78 are rejected.
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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/J.J.S./Examiner, Art Unit 1643
/Karen A. Canella/Primary Examiner, Art Unit 1643