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 57-77 are pending in the instant application and being examined on the merit.
Restriction Response
Applicant’s election of: 1) An effector molecule of phosphoramidate morpholino oligomer (PMO); 2) at least one single-domain antibody capable of binding to a binding site on a cell surface molecule of said cell (sdAb) of anti-CD71 sdAb; 3) a saponin, wherein the saponin comprises an aglycone core structure selected from quillaic acid and gypsogenin, wherein the saponin comprises a first saccharide chain bound to the C3 atom of an aglycone core structure, wherein the first saccharide chain comprises a glucuronic acid moiety, wherein the saponin is SO1861, in the reply filed on 12/25/2025 is acknowledged. Because applicant did not distinctly and specifically point out the supposed errors in the restriction requirement, the election has been treated as an election without traverse (MPEP § 818.01(a)).
Claim Objections
Claims 63 and 77 are objected to because of the following informalities: Regarding instant claim 63, the claim recites “acid,BNA” without a space in line 6.
Regarding instant claim 77, an indicated step ---c)--- is missing from the last line of the claim.
Appropriate correction is required.
Claim Rejections – 35 USC § 112(b)
The following is a quotation of 35 U.S.C. 112(b):
(b) CONCLUSION.—The specification shall conclude with one or more claims particularly pointing out and distinctly claiming the subject matter which the inventor or a joint inventor regards as the invention.
The following is a quotation of 35 U.S.C. 112 (pre-AIA ), second paragraph:
The specification shall conclude with one or more claims particularly pointing out and distinctly claiming the subject matter which the applicant regards as his invention.
Claims 57-77 are rejected under 35 U.S.C. 112(b) or 35 U.S.C. 112 (pre-AIA ), second paragraph, as being indefinite for failing to particularly point out and distinctly claim the subject matter which the inventor or a joint inventor (or for applications subject to pre-AIA 35 U.S.C. 112, the applicant), regards as the invention.
Regarding claim 57, the abbreviation “sdAb” for single domain antibody is not beside the definition. Thus, the metes and bounds are unclear because it is unclear is sdAb is an abbreviation for cell-surface molecule or single domain antibody based on placement of the abbreviation. Claims 58-77 further contain the unclear subject matter claimed and are also rejected.
Regarding instant claim 61, the claim recites selectable options as including an anti-CD71 sdAb or wherein the antibody is capable of binding to CD71. It is unclear what the differences are between these selectable options because both are binding the same target. Thus, the claim is indefinite and the metes and bounds are unclear.
Regarding instant claim 64, silencing a gene or targeting an mRNA, when present in a cell is claimed, wherein a gene is claimed as LDH, but LDH is a homo- or heterotetramer molecule, which is assembled by an association of two different subunits: A and B, wherein in humans, these polypeptide subunits are encoded by two structurally distinct genes: LDHA and LDHB. Thus, a gene of LDH is indefinite as to whether a gene of LDHA and/or LDHB is claimed.
To promote compact prosecution, the claim of LDH will be interpreted as LDHA and/or LDHB.
Regarding instant claim 77, Claim 77 recites the limitation "the binding site" in line 4. There is insufficient antecedent basis for this limitation in the claim. A binding site for the sdAb has not previously been introduced.
Claim Rejections – 35 USC § 112(a)
The following is a quotation of the first paragraph of 35 U.S.C. 112(a):
(a) IN GENERAL.—The specification shall contain a written description of the invention, and of the manner and process of making and using it, in such full, clear, concise, and exact terms as to enable any person skilled in the art to which it pertains, or with which it is most nearly connected, to make and use the same, and shall set forth the best mode contemplated by the inventor or joint inventor of carrying out the invention.
The following is a quotation of the first paragraph of pre-AIA 35 U.S.C. 112:
The specification shall contain a written description of the invention, and of the manner and process of making and using it, in such full, clear, concise, and exact terms as to enable any person skilled in the art to which it pertains, or with which it is most nearly connected, to make and use the same, and shall set forth the best mode contemplated by the inventor of carrying out his invention.
Claims 58 and 76 is rejected under 35 U.S.C. 112(a) or 35 U.S.C. 112 (pre-AIA ), first paragraph, as failing to comply with the written description requirement. The claim(s) contains subject matter which was not described in the specification in such a way as to reasonably convey to one skilled in the relevant art that the inventor or a joint inventor, or for applications subject to pre-AIA 35 U.S.C. 112, the inventor(s), at the time the application was filed, had possession of the claimed invention.
Regarding instant claim 58, the single domain antibodies are claimed that are a VH or VL, but the instant specification only teaches successful binding to a surface receptor and internalization with a VHH, with a VH or VL from any antibody. Thus, the Applicant does not have possession of VH or VL conjugates.
Regarding instant claim 76, a treatment of a disease is claimed, but the disease does not require a cell that expresses on its surface a binding site for the sdAb of the conjugate.
The purpose of the written description requirement is to ensure that the inventor had possession, at the time the invention was made, of the specific subject matter claimed. To satisfy the written description requirement, a patent specification must describe the claimed invention in sufficient detail that one skilled in the art can reasonably conclude that the inventor had possession of the claimed invention. See, e.g., Moba, B.V. v. Diamond Automation, Inc. , 325 F.3d 1306, 1319, 66 USPQ2d 1429, 1438 (Fed. Cir. 2003); Vas-Cath, Inc. v. Mahurkar, 935 F.2d at 1563, 19 USPQ2d at 1116.
Scope of the claimed genus
Instant claim 57 is drawn to a conjugate that comprises a sdAb, wherein the sdAb is selected from only a heavy chain variable region (VH) domain, light chain variable region (VL) domain, and a VHH. Thus, the claims are drawn to a genus of conjugates that are required to bind antigen yet only comprise a single VH or VL.
Instant claim 76, is drawn to a method for treating a disease in a patient in need thereof comprising administering to the patient an effective amount of a conjugate, wherein the disease does not require a cell that expresses on its surface a binding site for the sdAb of the conjugate.
State of the Relevant Art
The prior art has not taught a structure activity relationship that allows a sequence to be determined that requires only the VH or VL of an antibody for binding CD71. It is well established in the art that the formation of an intact antigen-binding site in an antibody usually requires the association of the complete heavy and light chain variable regions of a given antibody, each of which comprises three CDRs (or hypervariable regions) which provide the majority of the contact residues for the binding of the antibody to its target epitope. E.g., Almagro et. al., Front. Immunol. 2018; 8:1751, IDS reference (see Section “The IgG Molecule” in paragraph 1 and Figure 1). While affinity maturation techniques can result in differences in the CDRs of the antibody compared to its parental antibody (page 3 “The IgG Molecule, second and third paragraphs), those techniques involve trial-and-error testing and the changes that maintain or improve affinity are not predictable a priori. E.g., id., (page 6 ending paragraph onto page 7). Chiu ML et al. (Antibodies 2019 8, 55, 1-80, IDS reference) taught the antigen binding of antibodies often results in conformational changes in the contact surface areas of both the antibody and the antigen (page 5, first paragraph). Thus, the prediction of CDR binding to the epitope is difficult to predict. Single domain VHH antibodies are separate from individual VH or VL domains in an antibody. Wagner HJ et al. (Int J Mol Sci. 2018 19(11): 3444, IDS reference) taught in the context of nanobodies®, universal scaffolds have been identified, enabling the generation of robust or humanized VHH variants but, this strategy has only been applied to graft CDRs to acceptor frames obtained from animals of the same taxonomic family (page 2, last paragraph). Wagner taught the design of nanobody grafts with CDRs derived from conventional antibodies requires careful consideration, because both the heavy and light chain variable domain (VH and VL) form the antigen binding site and are involved in the recognition of the antigenic epitope (page 2, last paragraph). Furthermore, the framework plays an important role in CDR conformation and orientation and distinct framework residues often contribute directly to antigen binding (page 2, last paragraph).
Additionally, the prior art has taught single variable domains from species with VH and VL antibody pairing is distinct from other species with single domain only antibodies. (Holliger P et al. (Nature Biotechnology 2005 23 1126–1136, IDS reference). Holliger taught despite early excitement concerning the functional activity of single variable heavy domain antibodies, these antibody fragments rarely retained the affinity of the parent antibody and were also poorly soluble and often prone to aggregation (page 1127, left to right column bridging paragraph). Holliger taught while high affinity single variable like domain antibodies are present in camelid, as VhH, and shark, as V-NAR, domains, these single domain antibodies are structurally different, wherein each display long surface loops, often larger than for conventional murine and human antibodies, and are able to penetrate cavities in target antigens, such as enzyme active sites (for example, lysozyme) and canyons in viral and infectious disease biomarkers (page 1127, left to right column bridging paragraph). Holliger taught the structural changes are shown in Fig. 2, wherein superimposition of a human VH domain (Fig. 2b) and a single domain V-NAR(Fig. 2c) has a vastly different CDR3 that does not overlap structurally (Fig. 2d). Holliger taught unlike mouse VH domains, camelid VhH and shark V-NAR domains are in general soluble and can be produced as stable in vitro targeting reagents (page 1127, right column, second paragraph). Holliger did not describe single domain VL antibodies as successful. Thus, single-domain antibodies are not generally only a VH or VL and antibodies and require both the VH and VL, which are distinct from VHH.
The prior art does not teach a method wherein treatment of a disease with a sdAb targeting agent is effective wherein the disease does not require a cell that expresses on its surface a binding site for the sdAb of the conjugate for effective treatment. As described above, antigen-binding molecules specifically target epitopes of antigens.
Summary of Species disclosed in the original specification
MPEP § 2163 states that a “representative number of species” means that the species which are adequately described are representative of the entire genus. Thus, when there is substantial variation within the genus, one must describe a sufficient variety of species to reflect the variation within the genus.
It is noted that the specification does not disclose any antigen-binding regions that comprise only a VH or VL, which is distinguishable from a VHH; or a method wherein treatment of a disease with a sdAb targeting agent is effective wherein the disease does not require a cell that expresses on its surface a binding site for the sdAb of the conjugate for effective treatment.
Fig. 1A-E shows diagrams of a VHH conjugated toxin and VHH conjugated-SO1861 saponins separately and their expected results.
Fig. 2-7 show treatment of cells with a combination composition of a VHH targeting HER2 conjugated to SO1861 and a separate composition of an antibody or VHH conjugated cytotoxic agent.
Fig. 10 taught EGFR targeted HSP27 gene silencing with an anti-EGFR1 targeting cetuximab-(SO1861-Llinker-HSP27BNA oligonucleotide).
Fig. 18 shows the method of treatment of cancer in a mouse xenograft model wherein a combination of cetuximab-SO1861 and cetuximab-HSP27 BNA was administered to the subject and HSP27 transcription was decreased in the tumor. The antibodies tested include both a VH and VL.
One skilled in the art appreciates that traditional antibodies comprise two polypeptides, the light and heavy chains. The antigen-combining site of an antibody is a three-dimensional structure that fully comprises six CDRs, three each from the light and heavy chains. The amino acid sequences of the CDRs are hypervariable, as the amino acid residues contained within the CDRs determine much of the antibody’s antigen-binding specificity. Antibodies having less than all six CDRs that form the antigen binding site of a conventional antibody in their proper context of VHs and VLs do not describe the particularly identifying structural feature of the antibody that correlates with the antibody’s ability to bind antigen. Absent a description of the at least minimal structural features correlating with a functional ability to bind antigen which are shared by members of a genus commonly sharing this function, it is submitted that the skilled artisan could not immediately envision, recognize, or distinguish which VH and VL sequences should be combined such that a resultant antigen-binding domain is capable of binding antigen.
Accordingly, it is submitted that the skilled artisan could not immediately envision, recognize, or distinguish at least most of the members of the genus to which the claims are directed, and therefore the specification would not reasonably convey to the skilled artisan that Applicant was in possession of the claimed invention at the time the application was filed.
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 57-63 and 65-77 are rejected under 35 U.S.C. 103 as being unpatentable over US 2017/0281795 (Geall AJ et al.), WO 2020144233 (Cohen R et al.), Orellana E et al. (Mol Ther Nucleic Acids. 2019 Apr 11;16:505–518. doi: 10.1016/j.omtn.2019.04.003), Cinci M et al. (Target Oncol 2015 10(3):405-413), 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, IDS reference), King HD et al. (Bioconjugate Chem. 1999, 10, 2, 279–288, IDS reference), and US 2004/0242502 (Marciani D et al.).
Regarding instant claim 57 and 75-76, Geall taught an effective method of cancer treatment by silencing a protein in a cancer cell comprising administering a single pharmaceutical combination comprising a pharmaceutically acceptable excipient and:
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 instant claims 57-58, Geall taught the antibody as a single-domain antibody (sdAb) (page 1, [0019]). Regarding instant claims 57 and 74, Geall taught the endosomolytic moiety as an endosomolytic small molecule and an endosomolytic polymer (page 73, [0389]). Regarding instant claims 60-62, Geall taught targeting CD71 with an antibody siRNA conjugate (Fig. 54), wherein the CD71 targeting antibody siRNA conjugate effectively silenced HPRT expression (Fig. 86). Regarding instant claims 57 and 74, Geall taught antibody oligonucleotide conjugates comprising an siRNA oligonucleotide effector molecule, an antibody, and an endosome escape agent in a single conjugate (Fig. 1a) and antibody oligonucleotide conjugates comprising an siRNA oligonucleotide effector molecule, an antibody, and a polymeric molecule to which more than one endosome escape agent is bound (Fig. 1b). Regarding instant claim 63, Geall taught the effector molecule as a phosphorodiamidate morpholino oligomer (PMO) (page 59, paragraph 300). Geall taught the binding moiety as a nanobody® (also known as VHH) (page 1, paragraph 19).
Geall did not teach a pharmaceutical composition of a conjugate comprising a sdAb, siRNA effector and saponin of SO1861 conjugated together, but this is obvious in view of Cohen, Orellana, Cinci, Niewoehner, Weng, King, and Marciani.
Cohen taught a Transferrin receptor (CD71) targeted VHH conjugated to an siRNA (VHH-A and VHH-B) was effective at decreasing the protein level of the siRNA target (page 57, lines 1-15; page 58, lines 11-16; and Fig. 15D and 15H).
Regarding instant claims 57 and 70, Orellana taught a conjugate comprising a receptor targeting moiety, an oligonucleotide miRNA effector molecule and an endosomal escape moiety (Fig. 1), wherein the targeting moiety, oligonucleotide miRNA effector molecule and an endosomal escape moiety are conjugated to each other with a trivalent linker
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(Fig. 3), wherein ligand-targeted delivery of the endosomal escape moiety effectively promotes cargo escape from endosomes (Fig. 5) and effectively enhances effector miRNA activity in cells (Fig. 7A-C).
Regarding instant claims 60-62 and 76, Cinci taught a method for treating the disease of cancer wherein specific delivery of CD46, CD55, and CD59 (mCRP) siRNA via the transferrin receptor (CD71) to cancer cells that expressed high levels of CD71 selectively neutralized complement regulation in cancer cells and effectively enhanced specific antibody-mediated killing of tumor cells without affecting healthy bystander cells, which is a suited strategy to improve antibody-based cancer immunotherapy (abstract).
Regarding instant claims 59-62, 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 claims 57, 65-69, and 73, 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). Regarding instant claims 57, 65-69, and 73, Weng taught SO1861 augments the escape of the genetic cargo out of the intracellular compartments into the cytosol (abstract). Regarding instant claims 57, 65-69, and 73, 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). Regarding instant claims 57, 65-69, and 73, 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). Regarding instant claims 57, 65-69, and 73, 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). Regarding instant claim 77, Weng taught an effective in vitro method for transferring an oligonucleotide effector molecule from outside a 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 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).
Regarding instant claims 71-72, King taught immunoconjugates of monoclonal antibody BR96 and a doxorubicin payload employed a hydrazone linkage to C-13 of DOX and a thioether linkage to the antibody (page 279, left column, first paragraph). King taught hydrazone linkers of this nature are stable at normal physiological pH 7, but once internalized into the acidic compartment of endosomes/lysosomes, hydrolysis occurs liberating free doxorubicin payload (page 279, left column, first paragraph). King taught BR96-DOX conjugates produced antigen-specific antitumor activity and were more potent, were more active, and produced less systemic toxicity than unconjugated DOX in vivo (page 279, left column, first paragraph). King taught the branched series of BR96 conjugates demonstrated antigen-specific cytotoxicity, and were more potent in vitro than the single-chain conjugate on both a doxorubicin and antibody basis (abstract).
Regarding instant claims 66 and 72, 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 57-63 and 65-76, 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 comprising a pharmaceutically acceptable excipient and:
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:
Combine the antibody, oligonucleotide siRNA effector, and endosomal escape moiety into one conjugate as taught by Geall and with a trivalent linker wherein the antibody, oligonucleotide siRNA effector, and endosomal escape moiety are conjugated to each other in view of Orellana;
Exchange the antibody for a nanobody® (also known as VHH) as taught by Geall;
Exchange the siRNA target for CD46, CD55, and CD59 (mCRP) and combine the treatment with an antibody in view of Cinci;
Exchange the siRNA oligonucleotide effector for a phosphoramidate morpholino oligomer (PMO) as taught by Geall;
Exchange the EGFR targeting moiety with a bivalent CD71 targeted single domain antibody, wherein the bivalent antibodies are the same in view of Geall, Niewoehner,;
Exchange the endosomal escape molecule melittin for the endosomal escape molecule glycoside saponin SO1861 of Weng;
Include in the trivalent attachment linker: i) an acid sensitive hydrazone linker to the endosomal escape moiety and an acid sensitive hydrazone linker to the effector molecule in view of King; and ii) a single or branched polymer for linking the endosomal escape moiety in view of King and Geall;
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 a combined conjugate wherein the antibody, oligonucleotide siRNA effector, and endosomal escape moiety in one conjugate; and ii) Orellana taught a conjugate comprising a receptor targeting moiety, an oligonucleotide miRNA effector molecule and an endosomal escape moiety conjugated to each other with a trivalent linker, wherein ligand-targeted delivery of the endosomal escape moiety effectively promotes cargo escape from endosomes and effectively enhances effector miRNA activity in cells;
Geall taught the binding moiety as a nanobody® (also known as VHH);
Cinci taught a method for treating the disease of cancer wherein specific delivery of CD46, CD55, and CD59 (mCRP) siRNA through transferrin receptor (CD71) to cancer cells that expressed high levels of CD71 selectively neutralized complement regulation in cancer cells and effectively enhanced specific antibody-mediated killing of tumor cells without affecting healthy bystander cells, which is a suited strategy to improve antibody-based cancer immunotherapy;
Geall taught PMO as acceptable oligonucleotide effectors for antibody targeted treatment, wherein PMO are oligonucleotide effectors that target transcripts for the same purpose and would be expected to also be effective;
i) Geall taught the antibody as targeting CD71 with an antibody siRNA conjugate effectively and as a sdAb; ii) 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;
i) King taught immunoconjugates of monoclonal antibody BR96 and a doxorubicin payload with a single or branched polymer payload were effective in cancer cells and employed a hydrazone linkage to C-13 of DOX and a thioether linkage to the antibody, wherein hydrazone linkers of this nature are stable at normal physiological pH 7, but once internalized into the acidic compartment of endosomes/lysosomes, hydrolysis occurs liberating the free payload. 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, a hydrazone linker would release the SO1861 payload and oligonucleotide effector in the endosomes/lysosomes, where SO1861 has its effects; ii) Geall taught a polymeric molecule to which more than one endosome escape agent is bound;
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) The combined conjugate would target the same cells with the antibody, oligonucleotide siRNA effector, and endosomal escape moiety in one conjugate; and ii) Orellana taught a conjugate comprising a receptor targeting moiety, an oligonucleotide miRNA effector molecule and an endosomal escape moiety conjugated to each other with a trivalent linker, wherein ligand-targeted delivery of the endosomal escape moiety effectively promotes cargo escape from endosomes and effectively enhances effector miRNA activity in cells;
Exchanging the antibody for a nanobody® (also known as VHH) would still effectively target CD71 and CD71 conjugated oligonucleotides were known to be effective in view of Cohen;
Cinci taught a method for treating the disease of cancer wherein specific delivery of CD46, CD55, and CD59 (mCRP) siRNA through transferrin receptor (CD71) to cancer cells that expressed high levels of CD71 selectively neutralized complement regulation in cancer cells and effectively enhanced specific antibody-mediated killing of tumor cells without affecting healthy bystander cells, which is a suited strategy to improve antibody-based cancer immunotherapy;
PMO are oligonucleotide effectors that target transcripts for the same purpose and would be expected to also be effective;
i) Geall taught the antibody as targeting CD71 with an antibody siRNA conjugate effectively; ii) 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 and effector payloads 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 (instant claim 76) of a subject with a cancer overexpressing CD71, which is a cell surface receptor (instant claim 60), by silencing a protein in a cancer cell comprising administering a pharmaceutical composition comprising a pharmaceutically acceptable excipient (instant claim 75) and:
a conjugate comprising a bivalent VHH (instant claim 58) single domain CD71 targeted antibody (instant claims 61-62), a phosphoramidate morpholino oligomer (PMO) oligonucleotide effector molecule (instant claim 63) oligonucleotide targeting CD46, CD55, and CD59 (mCRP), and a single or polymeric molecule (instant claim 74) with modified SO1861 (instant claims 68-69) endosomal escape agents which has 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 65), wherein the VHH, PMO and SO1861 are covalently linked together with a trivalent linker that comprises hydrazone linkers attached to the SO1861 and PMO (instant claim 70), wherein the saponin comprises a first saccharide chain bound to the C3-beta-OH group of an aglycone core structure comprising a the glucuronic acid moiety (instant claims 67 and 73), wherein the SO1861 linker comprises an acid sensitive linker comprising a hydrazone bond linker connected at the C23 aldehyde of SO1861 (instant claims 66 and 71-72) 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 wherein the bivalent CD71 conjugate contains two VHH that are the same (instant claim 59) (instant claim 57).
Regarding instant claim 77, it would have been obvious for a person having ordinary skill in the art to take the composition from the method of Geall, Cohen, Orellana, Cinci, Niewoehner, Weng, King, and Marciani above of a pharmaceutical composition comprising:
a conjugate comprising a bivalent VHH single domain CD71 targeted antibody, a phosphoramidate morpholino oligomer (PMO) oligonucleotide effector molecule oligonucleotide targeting CD46, CD55, and CD59 (mCRP), and a single or polymeric molecule with modified SO1861 endosomal escape agents, wherein the VHH, PMO and SO1861 are covalently linked together with a trivalent linker that comprises hydrazone linkers attached to the SO1861 and PMO, wherein the saponin comprises a first saccharide chain bound to the C3-beta-OH group of an aglycone core structure comprising a the glucuronic acid moiety, wherein the SO1861 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 wherein the bivalent CD71 conjugate contains two VHH that are the same
– and: include the composition 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 composition comprising SO1861 and 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 conjugate comprising a bivalent VHH single domain CD71 targeted antibody, a phosphoramidate morpholino oligomer (PMO) oligonucleotide effector molecule oligonucleotide targeting CD46, CD55, and CD59 (mCRP), and a single or polymeric molecule with modified SO1861 endosomal escape agents, wherein the VHH, PMO and SO1861 are covalently linked together with a trivalent linker that comprises hydrazone linkers attached to the SO1861 and PMO, wherein the saponin comprises a first saccharide chain bound to the C3-beta-OH group of an aglycone core structure comprising a the glucuronic acid moiety, wherein the SO1861 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 wherein the bivalent CD71 conjugate contains two VHH that are the same;
providing a cell that expresses CD71 on the surface wherein the surface of the cell is targeted by the pharmaceutical compositions in a); and
contacting the cell of step b) in vitro with the pharmaceutical composition of step a),
and thus establishing the transfer of the oligonucleotide effector from outside the cell into the cell.
Claims 57-77 are rejected under 35 U.S.C. 103 as being unpatentable over US 2017/0281795 (Geall AJ et al.), WO 2020144233 (Cohen R et al.), Orellana E et al. (Mol Ther Nucleic Acids. 2019 Apr 11;16:505–518. doi: 10.1016/j.omtn.2019.04.003), Cinci M et al. (Target Oncol 2015 10(3):405-413), 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, IDS reference), King HD et al. (Bioconjugate Chem. 1999, 10, 2, 279–288, IDS reference), and US 2004/0242502 (Marciani D et al.) as applied to claims 57-63 and 65-77 above, and further in view of Xian Z-Y et al. (Tumour Biol. 2015 May 16;36(10):8093–8100.).
Geall, Cohen, Orellana, Cinci, Niewoehner, Weng, King, and Marciani are described above.
Regarding instant claim 64, Cinci further taught a method for treating the disease of cancer wherein specific delivery of CD46, CD55, and CD59 (mCRP) siRNA through transferrin receptor (CD71) to cancer cells that expressed high levels of CD71 such as DU145 selectively neutralized complement regulation in cancer cells and effectively enhanced specific antibody-mediated killing of tumor cells without affecting healthy bystander cells, which is a suited strategy to improve antibody-based cancer immunotherapy (abstract and Figs. 1-4).
Geall did not teach targeting an mRNA of LDHA, but this is obvious in view of Xian.
Xian taught decreased expression of LDHA by siRNA inhibited cell proliferation, migration, invasion, and promoted cell apoptosis of PC-3 and DU145 cells and indicated LDHA as a cancer therapeutic target (abstract and Fig. 2)
Regarding instant claim 64, it would have been obvious for a person having ordinary skill in the art to take the method of Geall, Cohen, Orellana, Cinci, Niewoehner, Weng, King, and Marciani above and in the pharmaceutical composition: – 1) exchange the CD46, CD55, and CD59 (mCRP) targeting effector oligonucleotide in the conjugate with an LDHA oligonucleotide conjugate of Xian.
This is obvious because: 1) Xian taught decreased expression of LDHA by siRNA inhibited cell proliferation, migration, invasion, and promoted cell apoptosis of PC-3 and DU145 cells and indicated LDHA as a cancer therapeutic target.
There is a reasonable expectation of success because: 1) Xian taught decreased expression of LDHA by siRNA inhibited cell proliferation, migration, invasion, and promoted cell apoptosis of PC-3 and DU145 cells and indicated LDHA as a cancer therapeutic target. A PMO targeting LDHA would thus be expected to block LDHA and effectively decrease cancer growth.
This would produce a method of cancer treatment of a subject with a cancer overexpressing CD71, which is a cell surface receptor, by silencing a protein in a cancer cell comprising administering a pharmaceutical composition comprising a pharmaceutically acceptable excipient and:
a conjugate comprising a bivalent VHH single domain CD71 targeted antibody, a phosphoramidate morpholino oligomer (PMO) oligonucleotide effector molecule oligonucleotide targeting LDHA, and a single or polymeric molecule with modified SO1861 endosomal escape agents, wherein the VHH, PMO and SO1861 are covalently linked together with a trivalent linker that comprises hydrazone linkers attached to the SO1861 and PMO, wherein the saponin comprises a first saccharide chain bound to the C3-beta-OH group of an aglycone core structure comprising a the glucuronic acid moiety, wherein the SO1861 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 wherein the bivalent CD71 conjugate contains two VHH that are the same.
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 57-77 are provisionally rejected on the ground of nonstatutory double patenting as being unpatentable over claims 57-80 of copending Application No. 18/012,729.
‘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 in copending claim 61, wherein the oligonucleotide is a nucleic acid in copending claim 63, wherein the oligonucleotide is a PMO 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.
‘729 did not teach a single embodiment of the elected species, but this is obvious in view of the separate claims of ‘729.
Regarding instant claims 57-74, it would have been obvious for a person having ordinary skill in the art to take copending claims 57-77 of ‘729 and combine them into one species to produce 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 (instant claim 58), wherein the sdAb is a VHH domain (instant claim 58), wherein the conjugate comprises at least two sdAb that are the same (instant claim 59), wherein the cell surface molecule is a cell surface receptor (instant claim 60), wherein the sdAb is an anti-CD71 antibody (instant claim 61-62), wherein the oligonucleotide effector is PMO (instant claim 63), wherein the oligonucleotide is capable of silencing LDH mRNA (instant claim 64), wherein the saponin comprises an aglycone core from quallaic acid and gypsogenin wherein C23 is an aldehyde or an acid sensitive bond (instant claims 65-66), wherein the saponin is SO1861 (instant claim 67-69), wherein the effector molecule is covalently bound to the sdAb and to the saponin (instant claim 70), wherein the linker covalently bound to the saponin is an acid sensitive linker to the stated C23 position (instant claims 71-72), 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 (instant claim 73), wherein the conjugate further comprises an oligomeric molecule to which more than one saponin is bound (instant claim 74).
This is obvious because: 1) copending claims 57-77 claim subject matter that overlaps with the instant application.
There is a reasonable expectation of success because: 1) all of the claimed components have overlapping subject matter and perform the same operation in the conjugate.
Regarding instant claims 75-76, it would have been obvious for a person having ordinary skill in the art to take the conjugate of copending claims 57-77 of ‘729 above – and: 1) use the conjugate in a method taught by copending claim 78 comprising treating a disease of cancer in a patient in need thereof by administering the conjugate, wherein the sdAb of the conjugate binds to a cell surface molecule present on the surface of the cells implicated in the disease and into which the effector molecule is intended to be delivered; and 2) include a pharmaceutically acceptable excipient in a pharmaceutically composition to dissolve the sdAb antibody conjugate.
This is obvious with a reasonable expectation of success because: 1) the copending method has overlapping subject matter that performs the same operation; and 2) administration of an sdAb conjugate would require a pharmaceutical composition and a pharmaceutically acceptable excipient.
Regarding instant claim 77, it would have been obvious for a person having ordinary skill in the art to take the conjugate of copending claims 57-77 of ‘729 above – and: 1) use the conjugate in a method taught by copending claim 80 comprising an in vitro method for transferring the conjugate of claim 57 from outside a cell to inside said cell comprising:(a) providing the conjugate of claim 57; (b) providing a cell which expresses on its surface a binding site for the sdAb of the conjugate; and (c) contacting the cell with the conjugate..
This is obvious with a reasonable expectation of success because: 1) the copending method has overlapping subject matter that performs the same operation.
This is a provisional nonstatutory double patenting rejection.
Claims 57-63 and 65-77 are provisionally rejected on the ground of nonstatutory double patenting as being unpatentable over:
claims 1-2, 5, 7, 11, 13-14, 17, 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; or
claims 1-2, 4, 9-12, 17, 21, 25, 32, 41, 44-45, 48, 51, 54, 56, 61, and 63 of copending Application No. 18/723,191,
in view of US 2017/0281795 (Geall AJ et al.), WO 2020144233 (Cohen R et al.), Orellana E et al. (Mol Ther Nucleic Acids. 2019 Apr 11;16:505–518. doi: 10.1016/j.omtn.2019.04.003), Cinci M et al. (Target Oncol 2015 10(3):405-413), 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, IDS reference), and King HD et al. (Bioconjugate Chem. 1999, 10, 2, 279–288, IDS reference).
‘104 copending claims 1-2, 5, 7, 11, 13-14, 17, 23-26, and 28-29 claim an oligonucleotide covalently conjugated to at least one saponin with a hydrazone linker, wherein the saponin is a bisdesmosidic 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 PMO 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 one or more saponins are conjugated in copending claim 24, wherein the conjugate further comprises an antibody that binds 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 a biologically active molecule covalently coupled to a 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 for covalently binding at least one endosomal escape enhancer bisdesmosidic triterpene saponin of a 12, 13-dehydrooleanne with an aldehyde function in position C-23 to a carrier molecule consisting of any of a proteinaceous molecule and a oligonucleotide, wherein the scaffold comprises a polymeric oligomeric structure and at least one of said bisdesmosidic triterpene saponin of a 12, 13-dehydrooleanne with an aldehyde function in position C-23, which is covalently coupled to the polymeric structure via a hydrazone bond in the C-23 position in copending claim 1, wherein the saponin 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 scaffold is a trifunctional linker comprising a covalent bond to the C-23 aldehyde position, a chemical group for covalent binding to a molecule, and a first chemical group for covalent bonding to the carrier in copending claim 16, wherein 1-128 glycoside molecules are in the scaffold in copending claim 17, wherein the carrier molecule is a VHH domain and an anti-CD71 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 PMO 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 of C which is a bisdesmosidic triterpene saponin with an aldehyde function in position C-23 that can be a polymer linked to an effector moiety A1 and a ligand B1 of a VHH in copending claim 1, wherein the ligand binds CD71 in copending claims 5-6, wherein the effector molecule is a PMO 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 claim 22; and wherein the therapeutic molecule is in a pharmaceutical composition with a pharmaceutically acceptable excipient in copending claim 35;
‘759 taught a method of treatment of cancer in a patient in need thereof comprising administering a pharmaceutical composition with a pharmaceutically acceptable excipient and a therapeutic molecule comprising a saponin of C which is a bisdesmosidic triterpene saponin with an aldehyde function in position C-23 that can be a polymer linked to an effector moiety A1 and a ligand B1 of a VHH in copending claim 38;
‘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 PMO;
‘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 PMO 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, wherein the at least one saponin is covalently bound to the cell-surface targeting molecule and at least one effector moiety via a linker in copending claim 28, and wherein the conjugate is in a pharmaceutical composition with a pharmaceutically acceptable excipient in copending claim 43.
‘191 copending claims 1-2, 4, 9-12, 17, 21, 25, 32, 41, 44-45, 48, 51, 54, 56, 61, and 63 claim a conjugate for delivery of an effector molecule from outside a cell into the cytosol, the conjugate comprising at least one effector molecule, at least saponin, and at least one sdAb, wherein the saponin, effector molecule, and at least one sdAb are covalently bound together, wherein the at least one saponin is quillaic acid, wherein the at least one effector molecule is an oligonucleotide and methods of treatment of cancer;
‘191 taught a conjugate for delivery of an effector molecule from outside a cell into the cytosol, the conjugate comprising at least one effector molecule, at least saponin, and at least one sdAb, wherein the saponin, effector molecule, and at least one sdAb are covalently bound together, wherein the at least one saponin is quillaic acid, wherein the at least one effector molecule is an oligonucleotide in copending claim 1, wherein the conjugate contains a bivalent VHH in copending claim 2, wherein the sdAb is an anti-CD71 in copending claim 4, wherein the sdAb is a VHH that binds CD71 in copending claim 11, wherein the conjugate comprises a multivalent antibody that is the same in copending claim 12, wherein the saponin comprises an aldehyde at position C-23 in copending claim 21, wherein the saponin is SO1861 in copending claim 25, wherein the effector molecule is PMO in copending claim 32, wherein the conjugate is in a pharmaceutical composition comprising a pharmaceutically acceptable excipient in copending claim 61, wherein the conjugate is administered in a method of cancer treatment to a patient in need thereof in copending claim 63.
While the claims of ‘104, ‘476, ‘759, ‘019, and ‘191 taught embodiments of a CD71 targeted antibody covalently conjugated to one or more of saponin SO1861 and an PMO, wherein the bond is a hydrazone or hydrazine bond subject to cleavage under acidic conditions, they did not teach a single embodiment of the elected species of a conjugate comprising an PMO effector, a CD71 sdAb, and a saponin SO1861, 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, but this is obvious in view of Geall, Cohen, Orellana, Cinci, Niewoehner, Weng, and King.
Regarding instant claim 57 and 75-76, Geall taught an effective method of cancer treatment by silencing a protein in a cancer cell comprising administering a single pharmaceutical combination comprising a pharmaceutically acceptable excipient and:
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 instant claims 57-58, Geall taught the antibody as a single-domain antibody (sdAb) (page 1, [0019]). Regarding instant claims 57 and 74, Geall taught the endosomolytic moiety as an endosomolytic small molecule and an endosomolytic polymer (page 73, [0389]). Regarding instant claims 60-62, Geall taught targeting CD71 with an antibody siRNA conjugate (Fig. 54), wherein the CD71 targeting antibody siRNA conjugate effectively silenced HPRT expression (Fig. 86). Regarding instant claims 57 and 74, Geall taught antibody oligonucleotide conjugates comprising an siRNA oligonucleotide effector molecule, an antibody, and an endosome escape agent in a single conjugate (Fig. 1a) and antibody oligonucleotide conjugates comprising an siRNA oligonucleotide effector molecule, an antibody, and a polymeric molecule to which more than one endosome escape agent is bound (Fig. 1b). Regarding instant claim 63, Geall taught the effector molecule as a phosphorodiamidate morpholino oligomer (PMO) (page 59, paragraph 300). Geall taught the binding moiety as a nanobody® (also known as VHH) (page 1, paragraph 19).
Cohen taught a Transferrin receptor (CD71) targeted VHH conjugated to an siRNA (VHH-A and VHH-B) was effective at decreasing the protein level of the siRNA target (page 57, lines 1-15; page 58, lines 11-16; and Fig. 15D and 15H).
Regarding instant claims 57 and 70, Orellana taught a conjugate comprising a receptor targeting moiety, an oligonucleotide miRNA effector molecule and an endosomal escape moiety (Fig. 1), wherein the targeting moiety, oligonucleotide miRNA effector molecule and an endosomal escape moiety are conjugated to each other with a trivalent linker
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(Fig. 3), wherein ligand-targeted delivery of the endosomal escape moiety effectively promotes cargo escape from endosomes (Fig. 5) and effectively enhances effector miRNA activity in cells (Fig. 7A-C).
Regarding instant claims 60-62 and 76, Cinci taught a method for treating the disease of cancer wherein specific delivery of CD46, CD55, and CD59 (mCRP) siRNA via the transferrin receptor (CD71) to cancer cells that expressed high levels of CD71 selectively neutralized complement regulation in cancer cells and effectively enhanced specific antibody-mediated killing of tumor cells without affecting healthy bystander cells, which is a suited strategy to improve antibody-based cancer immunotherapy (abstract).
Regarding instant claims 59-62, 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 claims 57, 65-69, and 73, 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). Regarding instant claims 57, 65-69, and 73, Weng taught SO1861 augments the escape of the genetic cargo out of the intracellular compartments into the cytosol (abstract). Regarding instant claims 57, 65-69, and 73, 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). Regarding instant claims 57, 65-69, and 73, 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). Regarding instant claims 57, 65-69, and 73, 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). Regarding instant claim 77, Weng taught an effective in vitro method for transferring an oligonucleotide effector molecule from outside a 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 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).
Regarding instant claims 71-72, King taught immunoconjugates of monoclonal antibody BR96 and a doxorubicin payload employed a hydrazone linkage to C-13 of DOX and a thioether linkage to the antibody (page 279, left column, first paragraph). King taught hydrazone linkers of this nature are stable at normal physiological pH 7, but once internalized into the acidic compartment of endosomes/lysosomes, hydrolysis occurs liberating free doxorubicin payload (page 279, left column, first paragraph). King taught BR96-DOX conjugates produced antigen-specific antitumor activity and were more potent, were more active, and produced less systemic toxicity than unconjugated DOX in vivo (page 279, left column, first paragraph). King taught the branched series of BR96 conjugates demonstrated antigen-specific cytotoxicity, and were more potent in vitro than the single-chain conjugate on both a doxorubicin and antibody basis (abstract).
Regarding instant claims 57-60, 62-74, and 76-77, it would have been obvious for a person having ordinary skill in the art to take copending claims:
1, 2, 7, 14, 17, 24, 26, and 28 of ‘104;
1, 5, 8, 11, 16-17, 19, 23, 24, and 26 of ‘476;
1, 5-6, 9, 13, 20, 22, 35 and 38 of ‘759;
1, 5, 9, 11, 17, 19, 24, 28, and 43 of ‘019, or
1-2, 4, 11-12, 21, 25, 32, 61, and 63 of '191,
of a CD71 targeted antibody covalently conjugated to one or more of saponin SO1861 and an PMO, 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 with a pharmaceutical excipient for cancer treatment in view of Geall;
Include a trivalent linker wherein the antibody, oligonucleotide siRNA effector, and endosomal escape moiety are conjugated to each other in view of Orellana;
Exchange the oligonucleotide target for CD46, CD55, and CD59 (mCRP) for cancer treatment and combine the treatment with an antibody in view of Cinci;
Use a VHH CD71 targeting moiety as taught by Cohen, ‘476, ‘759 and ‘019; and
Include a bivalent CD71 targeted single domain antibody, wherein the bivalent antibodies are the same in view of Niewoehner; and
Include in the trivalent attachment linker: i) an acid sensitive hydrazone linker to the SO1861 endosomal escape moiety at the C-23 aldehyde and an acid sensitive hydrazone linker to the effector molecule in view of King; and ii) a single or branched polymer for linking the endosomal escape moiety in view of King and Geall;
This is obvious because:
i) Geall taught a combined conjugate wherein the antibody, oligonucleotide siRNA effector, and endosomal escape moiety in one conjugate; and ii) Geall taught an effective method of cancer treatment by silencing a protein in a cancer cell comprising administering a single pharmaceutical combination comprising a pharmaceutically acceptable excipient 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 an endosomal escape moiety melittin;
Orellana taught a conjugate comprising a receptor targeting moiety, an oligonucleotide miRNA effector molecule and an endosomal escape moiety conjugated to each other with a trivalent linker, wherein ligand-targeted delivery of the endosomal escape moiety effectively promotes cargo escape from endosomes and effectively enhances effector miRNA activity in cells;
Cinci taught a method for treating the disease of cancer wherein specific delivery of CD46, CD55, and CD59 (mCRP) siRNA oligonucleotides through transferrin receptor (CD71) to cancer cells that expressed high levels of CD71 selectively neutralized complement regulation in cancer cells and effectively enhanced specific antibody-mediated killing of tumor cells without affecting healthy bystander cells, which is a suited strategy to improve antibody-based cancer immunotherapy;
‘476, ‘759 and ‘019 taught a VHH as an antibody targeting option and Cohen taught a Transferrin receptor (CD71) targeted VHH conjugated to an siRNA (VHH-A and VHH-B) was effective at decreasing the protein level of the siRNA target;
i) Geall taught the antibody as targeting CD71 with an antibody siRNA conjugate effectively and as a sdAb; ii) 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; and
i) King taught immunoconjugates of monoclonal antibody BR96 and a doxorubicin payload with a single or branched polymer payload were effective in cancer cells and employed a hydrazone linkage to C-13 of DOX and a thioether linkage to the antibody, wherein hydrazone linkers of this nature are stable at normal physiological pH 7, but once internalized into the acidic compartment of endosomes/lysosomes, hydrolysis occurs liberating the free payload. 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, a hydrazone linker would release the SO1861 payload and oligonucleotide effector in the endosomes/lysosomes, where SO1861 has its effects; ii) Geall and King taught a polymeric molecule to which more than one endosome escape agent is bound.
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;
Orellana taught a conjugate comprising a receptor targeting moiety, an oligonucleotide miRNA effector molecule and an endosomal escape moiety conjugated to each other with a trivalent linker, wherein ligand-targeted delivery of the endosomal escape moiety effectively promotes cargo escape from endosomes and effectively enhances effector miRNA activity in cells;
Cinci taught a method for treating the disease of cancer wherein specific delivery of CD46, CD55, and CD59 (mCRP) siRNA through transferrin receptor (CD71) to cancer cells that expressed high levels of CD71 selectively neutralized complement regulation in cancer cells and effectively enhanced specific antibody-mediated killing of tumor cells without affecting healthy bystander cells, which is a suited strategy to improve antibody-based cancer immunotherapy. Thus, oligonucleotides that target CD46, CD55, and CD59 (mCRP) would be effective;
Exchanging the antibody for a nanobody® (also known as VHH) would still effectively target CD71 and Cohen taught a Transferrin receptor (CD71) targeted VHH conjugated to an siRNA (VHH-A and VHH-B) was effective at decreasing the protein level of the siRNA target;
i) Geall taught the antibody as targeting CD71 with an antibody siRNA conjugate effectively; ii) 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;
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 and effector payloads in the endosomes/lysosomes, where SO1861 has its effects.
This would produce a method of cancer treatment (instant claim 76) of a subject with a cancer overexpressing CD71, which is a cell surface receptor (instant claim 60), by silencing a protein in a cancer cell comprising administering a pharmaceutical composition comprising a pharmaceutically acceptable excipient (instant claim 75) and:
a conjugate comprising a bivalent VHH (instant claim 58) single domain CD71 targeted antibody (instant claims 61-62), a phosphoramidate morpholino oligomer (PMO) oligonucleotide effector molecule (instant claim 63) oligonucleotide targeting CD46, CD55, and CD59 (mCRP), and a single or polymeric molecule (instant claim 74) with modified SO1861 (instant claims 68-69) endosomal escape agents which has 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 65), wherein the VHH, PMO and SO1861 are covalently linked together with a trivalent linker that comprises hydrazone linkers attached to the SO1861 and PMO (instant claim 70), wherein the saponin comprises a first saccharide chain bound to the C3-beta-OH group of an aglycone core structure comprising a the glucuronic acid moiety (instant claims 67 and 73), wherein the SO1861 linker comprises an acid sensitive linker comprising a hydrazone bond linker connected at the C23 aldehyde of SO1861 (instant claims 66 and 71-72) 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 wherein the bivalent CD71 conjugate contains two VHH that are the same (instant claim 59) (instant claim 57).
Regarding instant claim 77, 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, ‘019, or ‘191 and Geall, Cohen, Orellana, Cinci, Niewoehner, Weng, and King above of a pharmaceutical composition comprising:
a conjugate comprising a bivalent VHH single domain CD71 targeted antibody, a phosphoramidate morpholino oligomer (PMO) oligonucleotide effector molecule oligonucleotide targeting CD46, CD55, and CD59 (mCRP), and a single or polymeric molecule with modified SO1861 endosomal escape agents, wherein the VHH, PMO and SO1861 are covalently linked together with a trivalent linker that comprises hydrazone linkers attached to the SO1861 and PMO, wherein the saponin comprises a first saccharide chain bound to the C3-beta-OH group of an aglycone core structure comprising a the glucuronic acid moiety, wherein the SO1861 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 wherein the bivalent CD71 conjugate contains two VHH that are the same
– and: include the composition 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 composition comprising SO1861 and 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 conjugate comprising a bivalent VHH single domain CD71 targeted antibody, a phosphoramidate morpholino oligomer (PMO) oligonucleotide effector molecule oligonucleotide targeting CD46, CD55, and CD59 (mCRP), and a single or polymeric molecule with modified SO1861 endosomal escape agents, wherein the VHH, PMO and SO1861 are covalently linked together with a trivalent linker that comprises hydrazone linkers attached to the SO1861 and PMO, wherein the saponin comprises a first saccharide chain bound to the C3-beta-OH group of an aglycone core structure comprising a the glucuronic acid moiety, wherein the SO1861 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 wherein the bivalent CD71 conjugate contains two VHH that are the same;
providing a cell that expresses CD71 on the surface wherein the surface of the cell is targeted by the pharmaceutical compositions in a); and
contacting the cell of step b) in vitro with the pharmaceutical composition of step a),
and thus establishing the transfer of the oligonucleotide effector from outside the cell into the cell.
This is a provisional nonstatutory double patenting rejection.
Claims 57-77 are provisionally rejected on the ground of nonstatutory double patenting as being unpatentable over:
claims 1-2, 5, 7, 11, 13-14, 17, 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, or
claims 1-2, 4, 9-12, 17, 21, 25, 32, 41, 44-45, 48, 51, 54, 56, 61, and 63 of copending Application No. 18/723,191,
in view of US 2017/0281795 (Geall AJ et al.), WO 2020144233 (Cohen R et al.), Orellana E et al. (Mol Ther Nucleic Acids. 2019 Apr 11;16:505–518. doi: 10.1016/j.omtn.2019.04.003), Cinci M et al. (Target Oncol 2015 10(3):405-413), 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, IDS reference), King HD et al. (Bioconjugate Chem. 1999, 10, 2, 279–288, IDS reference), and Xian Z-Y et al. Inhibition of LDHA suppresses tumor progression in prostate cancer. (Tumour Biol. 2015 May 16;36(10):8093–8100.).
The claims of ‘104, ‘476, 759, ‘019, or ‘191 and Geall, Orellana, Cinci, Niewoehner, Weng, and King teach the limitations of instant claims 57-63 and 65-77 as set forth above.
‘104, ‘476, 759, ‘019, or ‘191, and Geall, Orellana, Cinci, Niewoehner, Weng, and King are described above.
Regarding instant claim 64, Cinci further taught a method for treating the disease of cancer wherein specific delivery of CD46, CD55, and CD59 (mCRP) siRNA through transferrin receptor (CD71) to cancer cells that expressed high levels of CD71 such as DU145 selectively neutralized complement regulation in cancer cells and effectively enhanced specific antibody-mediated killing of tumor cells without affecting healthy bystander cells, which is a suited strategy to improve antibody-based cancer immunotherapy (abstract and Figs. 1-4).
‘104, ‘476, 759, ‘019, and ‘191 did not teach targeting an mRNA of LDHA, but this is obvious in view of Xian.
Xian taught decreased expression of LDHA by siRNA inhibited cell proliferation, migration, invasion, and promoted cell apoptosis of PC-3 and DU145 cells and indicated LDHA as a cancer therapeutic target (abstract and Fig. 2)
Regarding instant claim 64, it would have been obvious for a person having ordinary skill in the art to take the method of ‘104, ‘476, 759, ‘019, or ‘191 and Geall, Orellana, Cinci, Niewoehner, Weng, and King above and in the pharmaceutical composition: – 1) exchange the CD46, CD55, and CD59 (mCRP) targeting effector oligonucleotide in the conjugate with an LDHA oligonucleotide conjugate of Xian.
This is obvious because: 1) Xian taught decreased expression of LDHA by siRNA inhibited cell proliferation, migration, invasion, and promoted cell apoptosis of PC-3 and DU145 cells and indicated LDHA as a cancer therapeutic target.
There is a reasonable expectation of success because: 1) Xian taught decreased expression of LDHA by siRNA inhibited cell proliferation, migration, invasion, and promoted cell apoptosis of PC-3 and DU145 cells and indicated LDHA as a cancer therapeutic target. A PMO targeting LDHA would thus be expected to block LDHA and effectively decrease cancer growth.
This is a provisional nonstatutory double patenting rejection.
Claims 57-74 and 76-78 are provisionally rejected on the ground of nonstatutory double patenting as being unpatentable over:
claims 57-59, 61, 64-65, 67-78 of copending Application No.18/012,754;
claims 41-57 of copending Application No.19/044,661;
claims 57-65 and 67-78 of copending Application No. 18/012,741; or
claims 1-57 of copending Application No. 18/262994,
in view of in view of Orellana E et al. (Mol Ther Nucleic Acids. 2019 Apr 11;16:505–518. doi: 10.1016/j.omtn.2019.04.003), Cinci M et al. (Target Oncol 2015 10(3):405-413), Weng A et al. (Journal of Controlled Release 2015 206 75-90, IDS reference), and King HD et al. (Bioconjugate Chem. 1999, 10, 2, 279–288, IDS reference).
‘754 copending claims 57-59, 61, 64-65, 67-78 claim a pharmaceutical combination and a pharmaceutically acceptable excipient comprising: a first conjugate comprising an effector molecule and a single-domain antibody for binding to a first cell-surface molecule (sdAb), wherein the sdAb is a VHH domain that can bind CD71, wherein the effector molecule and the sdAb are covalently linked to each other and wherein the effector molecule comprises or consists of a nucleic acid or a xeno nucleic acid;
a saponin and/or a modified saponin, wherein the saponin is a monodesmosidic triterpene glycoside or a bidesmosidic triterpene glycoside, wherein the modified saponin is a monodesmosidic triterpene glycoside or a bidesmosidic triterpene glycoside comprising a modified aglycone core structure being a modified quillaic acid or a modified gypsogenin wherein the aldehyde group in position C-23 of quillaic acid or of gypsogenin, respectively, is chemically modified by transformation into a hydrazone bond and methods;
‘754 taught a pharmaceutical combination and a pharmaceutically acceptable excipient comprising: a first conjugate comprising an effector molecule and a single-domain antibody for binding to a first cell-surface molecule (sdAb), wherein the sdAb is a VHH domain that can bind CD71, wherein the effector molecule and the sdAb are covalently linked to each other and wherein the effector molecule comprises or consists of a nucleic acid or a xeno nucleic acid; a saponin and/or a modified saponin, wherein the saponin is a monodesmosidic triterpene glycoside or a bidesmosidic triterpene glycoside, wherein the modified saponin is a monodesmosidic triterpene glycoside or a bidesmosidic triterpene glycoside comprising a modified aglycone core structure being a modified quillaic acid or a modified gypsogenin wherein the aldehyde group in position C-23 of quillaic acid or of gypsogenin, respectively, is chemically modified by transformation into a hydrazone bond in copending claim 57, wherein the first conjugate and the saponin and/or the modified saponin are provided in a single pharmaceutical composition in copending claim 59, wherein the first conjugate comprises two sdAbs that are the same in copending claim 61, wherein the effector molecule is an PMO in copending claim 64, wherein the saponin is a modified quillaic acid wherein the aldehyde group in position C-23 of quillaic acid is chemically modified by transformation into a hydrazone bond in copending claim 67, wherein the hyrazone bond comprised by the modified saponin was obtained by chemically modifying the aldehyde group in position C-23 of quillaic acid or of gypsogenin:- through reaction with N-s-maleimidocaproic acid hydrazide (EMCH) in copending claim 69, wherein the first saccharide chain comprising the glucuronic acid unit either comprises a carboxyl group on the glucuronic acid unit or comprises an amide bond in copending claim 70 wherein the saponin is SO1861 in copending claims 71-74, wherein the modified saponin is comprised within a second conjugate, which comprises a binding molecule that is covalently linked in copending claim 77, wherein the modified saponin is covalently bound to the binding molecule via a hydrazone bond at C-23 in copending claim 78. ‘754 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.
‘754 taught an in vitro method for transferring an effector molecule comprising or consisting of a nucleic acid or a xeno nucleic acid from outside a cell to inside said cell comprising: a) providing the first conjugate of the pharmaceutical combination above; b) providing the saponin or the modified saponin of the pharmaceutical combination of claim 57; c) providing a cell which expresses the first cell-surface molecule as recognized by the sdAb of the first conjugate; and d) contacting the cell in vitro with the first conjugate and the saponin or the modified saponin, therewith establishing the transfer of the effector molecule from outside the cell into said cell in copending claim 76.
‘661 copending claims 41-57 taught a methods 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 covalently bound saponin and 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 and a pharmaceutically acceptable excipient.
‘661 copending claims 41-57 taught a methods 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 covalently bound saponin and 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 and 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 via an aldehyde function in the C-23 position in the saponin via at least one cleavable linker 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 and second binding site comprise an anti-CD71 antibody in copending claim 45, wherein the effector moiety is a PMO 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 human cells 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.
‘741 copending claims 57-65 and 67-78 claim a pharmaceutical combination comprising – a first conjugate comprising a sdAb that binds to a first cell surface molecule wherein the sdAb is covalently linked to an effector molecule comprising an oligonucleotide; and a second conjugate comprising a binding molecule to a second cell surface molecule covalently linked to a mono-desmosidic triterpene glycoside comprising a quillaic acid or gypsogenin with an aldehyde or acid-sensitive bond at position C23 of the aglycone core wherein the first and second cell surface molecules are present on the same cell.
‘741 taught a pharmaceutical combination comprising – a first conjugate comprising a sdAb that binds to a first cell surface molecule wherein the sdAb is covalently linked to an effector molecule comprising an oligonucleotide; and a second conjugate comprising a binding molecule to a second cell surface molecule covalently linked to a mono-desmosidic triterpene glycoside comprising a quillaic acid or gypsogenin with an aldehyde or acid-sensitive bond at position C23 of the aglycone core wherein the first and second cell surface molecules are present on the same cell in copending claim 57, wherein the first and second conjugate are provided in a single pharmaceutical composition copending claim 59, wherein the sdAb is a VHH domain in copending claim 60, wherein the conjugate comprises at least two sdAbs, wherein two sdAbs are the same in copending claim 61, wherein the sdAb is an anti-CD71 sdAb in copending claim 62-63, wherein the oligonucleotide is a PMO in copending claim 64, wherein the saponin is SO1861 in copending claims 68-69, wherein the binding domain of the second molecule is an immunoglobin or binding fragment of domain of an immunoglobin in copending claim 70, wherein the saponin is covalently linked to the binding molecule via a linker in copending claim 72, wherein the linker is an acid sensitive linker in copending claim 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, wherein the second conjugate comprises more than one molecule of saponin derivative and further comprises an oligomeric molecule to which more than one molecule of saponin is covalently bound in copending claim 76,
‘741 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 of copending claim 57;
‘741 taught an in vitro method for transferring an oligonucleotide from outside a cell to inside said cell comprising:
providing the first conjugate of copending claim 57;
providing the second conjugate of copending claim 57
providing a cell which expresses on its surface the binding site for the sdAb of the first conjugate; and second cell surface molecule;
contacting the cell in step iii) in vitro with the first conjugate of step i) and second conjugate of step ii), therewith establishing the transfer of the effector molecule from outside the cell into said cell in copending claim 78;
‘994 copending claims 1-57 claim a saponin conjugates comprising a first proteinaceous comprising a cell-surface molecule comprising a first binding site for binding a first epitope of a first cell surface molecule covalently bound to a saponin comprising an aglycone core structure;
‘994 taught a saponin conjugates comprising a first proteinaceous comprising a cell-surface molecule comprising a first binding site for binding a first epitope of a first cell surface molecule covalently bound to a saponin comprising an aglycone core structure in copending claim 1, wherein the saponin is a mono-desmosidic triterpene glycoside with an aldehyde in position C-23 in copending claim 2, wherein the saponin conjugate is SO1861 in copending claims 10-12, wherein formula XII taught a C-23 position hydrazone linker attachment to the proteinaceous molecule in copending claim 15, wherein multiple saponins are conjugated in copending claims 19-20, wherein the first binding site of the proteinaceous molecule is a VHH in copending claims 22 and 24, wherein the cell surface receptor the proteinaceous molecule binds is CD71 in copending claim 26, wherein the proteinaceous molecule is a sdAb of an anti-CD71 antibody in copending claim 27, wherein the saponin conjugate is in a pharmaceutical composition and a pharmaceutically acceptable excipient in copending claim 28, wherein the first a saponin conjugate composition is in a pharmaceutical combination further comprises a second pharmaceutical composition comprising a covalently bound conjugate of a third proteinaceous molecule and an effector moiety wherein the binding site of the proteinaceous molecules are the same in copending claim 31, wherein the third proteinaceous molecule is an anti-CD71 VHH in copending claim 32, wherein the effector moiety is PMO in copending claim 35, wherein the first and second pharmaceutical composition are used in a treatment of cancer in copending claim 50.
While the claims of ‘754, ‘661, ‘741, and ‘994 taught embodiments of a sdAb CD71 VHH targeted antibody covalently conjugated to one or more of saponin SO1861 at C23 via a hydrazone bond and an antibody conjugated to an PMO oligonucleotide, they did not teach the conjugates as a single conjugate on a CD71 targeting VHH conjugated to one or more of saponin SO1861 at C23 via a hydrazone bond and a PMO effector oligonucleotide, but this is obvious in view of Orellana, Cinci, Weng, and King.
Regarding instant claims 57 and 70, Orellana taught a conjugate comprising a receptor targeting moiety, an oligonucleotide miRNA effector molecule and an endosomal escape moiety (Fig. 1), wherein the targeting moiety, oligonucleotide miRNA effector molecule and an endosomal escape moiety are conjugated to each other with a trivalent linker
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(Fig. 3), wherein ligand-targeted delivery of the endosomal escape moiety effectively promotes cargo escape from endosomes (Fig. 5) and effectively enhances effector miRNA activity in cells (Fig. 7A-C).
Regarding instant claims 60-62 and 76, Cinci taught a method for treating the disease of cancer wherein specific delivery of CD46, CD55, and CD59 (mCRP) siRNA via the transferrin receptor (CD71) to cancer cells that expressed high levels of CD71 selectively neutralized complement regulation in cancer cells and effectively enhanced specific antibody-mediated killing of tumor cells without affecting healthy bystander cells, which is a suited strategy to improve antibody-based cancer immunotherapy (abstract).
Regarding instant claims 57, 65-69, and 73, 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). Regarding instant claims 57, 65-69, and 73, Weng taught SO1861 augments the escape of the genetic cargo out of the intracellular compartments into the cytosol (abstract). Regarding instant claims 57, 65-69, and 73, 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). Regarding instant claims 57, 65-69, and 73, 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). Regarding instant claims 57, 65-69, and 73, 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). Regarding instant claim 77, Weng taught an effective in vitro method for transferring an oligonucleotide effector molecule from outside a 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 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).
Regarding instant claims 71-72, King taught immunoconjugates of monoclonal antibody BR96 and a doxorubicin payload employed a hydrazone linkage to C-13 of DOX and a thioether linkage to the antibody (page 279, left column, first paragraph). King taught hydrazone linkers of this nature are stable at normal physiological pH 7, but once internalized into the acidic compartment of endosomes/lysosomes, hydrolysis occurs liberating free doxorubicin payload (page 279, left column, first paragraph). King taught BR96-DOX conjugates produced antigen-specific antitumor activity and were more potent, were more active, and produced less systemic toxicity than unconjugated DOX in vivo (page 279, left column, first paragraph). King taught the branched series of BR96 conjugates demonstrated antigen-specific cytotoxicity, and were more potent in vitro than the single-chain conjugate on both a doxorubicin and antibody basis (abstract).
Regarding instant claims 57-74, and 76-77, it would have been obvious for a person having ordinary skill in the art to take copending claims:
57, 59, 61, 64, 67, 69, 70-74, 77, and 78 of '754;
41, 43-46, 50, 53, and 55 of '661;
57, 59-64, 68-70, 72-74, and 76 of '741; and
1-2, 10-12, 15, 19-20, 22, 24, 26-28, 31-32, and 35 of '994
of a method of treating a disease of cancer in a patient in need thereof comprising administering to the patient an effective amount of a pharmaceutical combination of a pharmaceutically acceptable excipient comprising: a first conjugate comprising two sdAb CD71 targeting VHH that are the same conjugated to a PMO effector nucleotide, wherein the effector molecule and the sdAb are covalently linked to each other; and
a second conjugate, which comprises a binding molecule that is covalently linked via a hydrazone bond at C-23 of the saponin SO1081, wherein the saponin is a modified quillaic acid wherein the aldehyde group in position C-23 of quillaic acid is chemically modified by transformation into a hydrazone bond, wherein the hyrazone bond comprised by the modified saponin was obtained by chemically modifying the aldehyde group in position C-23 of quillaic acid or of gypsogenin – and:
Include a trivalent linker wherein the antibody, oligonucleotide siRNA effector, and endosomal escape moiety are conjugated to each other in view of Orellana or ‘994;
Include in the trivalent attachment linker: i) an acid sensitive hydrazone linker to the SO1861 endosomal escape moiety at the C-23 aldehyde and an acid sensitive hydrazone linker to the effector molecule in view of King; and ii) a single or branched polymer for linking the endosomal escape moiety in view of King, or ‘994; and
Exchange the oligonucleotide target for CD46, CD55, and CD59 (mCRP) for cancer treatment and combine the treatment with an antibody in view of Cinci.
This is obvious because:
Orellana taught a conjugate comprising a receptor targeting moiety, an oligonucleotide miRNA effector molecule and an endosomal escape moiety conjugated to each other with a trivalent linker, wherein ligand-targeted delivery of the endosomal escape moiety effectively promotes cargo escape from endosomes and effectively enhances effector miRNA activity in cells; and
i) King taught immunoconjugates of monoclonal antibody BR96 and a doxorubicin payload with a single or branched polymer payload were effective in cancer cells and employed a hydrazone linkage to C-13 of DOX and a thioether linkage to the antibody, wherein hydrazone linkers of this nature are stable at normal physiological pH 7, but once internalized into the acidic compartment of endosomes/lysosomes, hydrolysis occurs liberating the free payload. 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, a hydrazone linker would release the SO1861 payload and oligonucleotide effector in the endosomes/lysosomes, where SO1861 has its effects; ii) King taught a polymeric molecule to which more than one endosome escape agent is bound; and
Cinci taught a method for treating the disease of cancer wherein specific delivery of CD46, CD55, and CD59 (mCRP) siRNA through transferrin receptor (CD71) to cancer cells that expressed high levels of CD71 selectively neutralized complement regulation in cancer cells and effectively enhanced specific antibody-mediated killing of tumor cells without affecting healthy bystander cells, which is a suited strategy to improve antibody-based cancer immunotherapy. Thus, oligonucleotides that target CD46, CD55, and CD59 (mCRP) would be effective.
There is a reasonable expectation of success because:
Orellana taught a conjugate comprising a receptor targeting moiety, an oligonucleotide miRNA effector molecule and an endosomal escape moiety conjugated to each other with a trivalent linker, wherein ligand-targeted delivery of the endosomal escape moiety effectively promotes cargo escape from endosomes and effectively enhances effector miRNA activity in cells;
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 and effector payloads in the endosomes/lysosomes, where SO1861 has its effects; and
Cinci taught a method for treating the disease of cancer wherein specific delivery of CD46, CD55, and CD59 (mCRP) siRNA through transferrin receptor (CD71) to cancer cells that expressed high levels of CD71 selectively neutralized complement regulation in cancer cells and effectively enhanced specific antibody-mediated killing of tumor cells without affecting healthy bystander cells, which is a suited strategy to improve antibody-based cancer immunotherapy. Thus, oligonucleotides that target CD46, CD55, and CD59 (mCRP) would be effective.
This would produce a method of cancer treatment (instant claim 76) of a subject with a cancer overexpressing CD71, which is a cell surface receptor (instant claim 60), by silencing a protein in a cancer cell comprising administering a pharmaceutical composition comprising a pharmaceutically acceptable excipient (instant claim 75) and:
a conjugate comprising a bivalent VHH (instant claim 58) single domain CD71 targeted antibody (instant claims 61-62), a phosphoramidate morpholino oligomer (PMO) oligonucleotide effector molecule (instant claim 63) oligonucleotide targeting CD46, CD55, and CD59 (mCRP), and a single or polymeric molecule (instant claim 74) with modified SO1861 (instant claims 68-69) endosomal escape agents which has 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 65), wherein the VHH, PMO and SO1861 are covalently linked together with a trivalent linker that comprises hydrazone linkers attached to the SO1861 and PMO (instant claim 70), wherein the saponin comprises a first saccharide chain bound to the C3-beta-OH group of an aglycone core structure comprising a the glucuronic acid moiety (instant claims 67 and 73), wherein the SO1861 linker comprises an acid sensitive linker comprising a hydrazone bond linker connected at the C23 aldehyde of SO1861 (instant claims 66 and 71-72) 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 wherein the bivalent CD71 conjugate contains two VHH that are the same (instant claim 59) (instant claim 57).
Regarding instant claim 77, it would have been obvious for a person having ordinary skill in the art to take the composition from the method of '754, '661, '741, or '994, Geall, Cohen, Orellana, Cinci, Niewoehner, Weng, and King above of a pharmaceutical composition comprising:
a conjugate comprising a bivalent VHH single domain CD71 targeted antibody, a phosphoramidate morpholino oligomer (PMO) oligonucleotide effector molecule oligonucleotide targeting CD46, CD55, and CD59 (mCRP), and a single or polymeric molecule with modified SO1861 endosomal escape agents, wherein the VHH, PMO and SO1861 are covalently linked together with a trivalent linker that comprises hydrazone linkers attached to the SO1861 and PMO, wherein the saponin comprises a first saccharide chain bound to the C3-beta-OH group of an aglycone core structure comprising a the glucuronic acid moiety, wherein the SO1861 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 wherein the bivalent CD71 conjugate contains two VHH that are the same
– and: include the composition 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 composition comprising SO1861 and 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.
Claims 57-77 are provisionally rejected on the ground of nonstatutory double patenting as being unpatentable over:
claims 57-59, 61, 64-65, 67-78 of copending Application No.18/012,754;
claims 41-57 of copending Application No.19/044,661;
claims 57-65 and 67-78 of copending Application No. 18/012,741; or
claims 1-57 of copending Application No. 18/262,994,
in view of in view of Orellana E et al. (Mol Ther Nucleic Acids. 2019 Apr 11;16:505–518. doi: 10.1016/j.omtn.2019.04.003), Cinci M et al. (Target Oncol 2015 10(3):405-413), Weng A et al. (Journal of Controlled Release 2015 206 75-90, IDS reference), and King HD et al. (Bioconjugate Chem. 1999, 10, 2, 279–288, IDS reference), and Xian Z-Y et al. Inhibition of LDHA suppresses tumor progression in prostate cancer. (Tumour Biol. 2015 May 16;36(10):8093–8100.).
The claims of ‘754, ‘661, ‘741, or ‘994 and Orellana, Cinci, Weng, and King teach the limitations of instant claims 57-63 and 65-77 as set forth above.
‘104, ‘476, 759, or ‘019 and Orellana, Cinci, Weng, and King are described above.
Regarding instant claim 64, Cinci further taught a method for treating the disease of cancer wherein specific delivery of CD46, CD55, and CD59 (mCRP) siRNA through transferrin receptor (CD71) to cancer cells that expressed high levels of CD71 such as DU145 selectively neutralized complement regulation in cancer cells and effectively enhanced specific antibody-mediated killing of tumor cells without affecting healthy bystander cells, which is a suited strategy to improve antibody-based cancer immunotherapy (abstract and Figs. 1-4).
‘754, ‘661, or ‘741 did not teach targeting an mRNA of LDHA, but this is obvious in view of Xian.
Xian taught decreased expression of LDHA by siRNA inhibited cell proliferation, migration, invasion, and promoted cell apoptosis of PC-3 and DU145 cells and indicated LDHA as a cancer therapeutic target (abstract and Fig. 2)
Regarding instant claim 64, it would have been obvious for a person having ordinary skill in the art to take the method of ‘754, ‘661, ‘741, or ‘994 and Geall, Orellana, Cinci, Niewoehner, Weng, and King above and in the pharmaceutical composition: – 1) exchange the CD46, CD55, and CD59 (mCRP) targeting effector oligonucleotide in the conjugate with an LDHA oligonucleotide conjugate of Xian.
This is obvious because: 1) Xian taught decreased expression of LDHA by siRNA inhibited cell proliferation, migration, invasion, and promoted cell apoptosis of PC-3 and DU145 cells and indicated LDHA as a cancer therapeutic target.
There is a reasonable expectation of success because: 1) Xian taught decreased expression of LDHA by siRNA inhibited cell proliferation, migration, invasion, and promoted cell apoptosis of PC-3 and DU145 cells and indicated LDHA as a cancer therapeutic target. A PMO targeting LDHA would thus be expected to block LDHA and effectively decrease cancer growth.
This is a provisional nonstatutory double patenting rejection.
Claims 57-63 and 65-77 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.), WO 2020144233 (Cohen R et al.), Orellana E et al. (Mol Ther Nucleic Acids. 2019 Apr 11;16:505–518. doi: 10.1016/j.omtn.2019.04.003), Cinci M et al. (Target Oncol 2015 10(3):405-413), 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, IDS reference), King HD et al. (Bioconjugate Chem. 1999, 10, 2, 279–288, IDS reference), and US 2004/0242502 (Marciani D et al.).
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 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 PMO 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.
The claims of ‘760, ‘193, or ‘163 taught embodiments of an antibody targeted saponin and/or oligonucleotide, wherein the saponin is SO1861, but does not teach a single embodiment of the elected species of a conjugate comprising an PMO effector, a CD71 sdAb, and a saponin SO1861, 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, but this is obvious in view of Geall, Cohen, Orellana, Cinci, Niewoehner, Weng, and King.
Regarding instant claim 57 and 75-76, Geall taught an effective method of cancer treatment by silencing a protein in a cancer cell comprising administering a single pharmaceutical combination comprising a pharmaceutically acceptable excipient and:
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 instant claims 57-58, Geall taught the antibody as a single-domain antibody (sdAb) (page 1, [0019]). Regarding instant claims 57 and 74, Geall taught the endosomolytic moiety as an endosomolytic small molecule and an endosomolytic polymer (page 73, [0389]). Regarding instant claims 60-62, Geall taught targeting CD71 with an antibody siRNA conjugate (Fig. 54), wherein the CD71 targeting antibody siRNA conjugate effectively silenced HPRT expression (Fig. 86). Regarding instant claims 57 and 74, Geall taught antibody oligonucleotide conjugates comprising an siRNA oligonucleotide effector molecule, an antibody, and an endosome escape agent in a single conjugate (Fig. 1a) and antibody oligonucleotide conjugates comprising an siRNA oligonucleotide effector molecule, an antibody, and a polymeric molecule to which more than one endosome escape agent is bound (Fig. 1b). Regarding instant claim 63, Geall taught the effector molecule as a phosphorodiamidate morpholino oligomer (PMO) (page 59, paragraph 300). Geall taught the binding moiety as a nanobody® (also known as VHH) (page 1, paragraph 19).
Cohen taught a Transferrin receptor (CD71) targeted VHH conjugated to an siRNA (VHH-A and VHH-B) was effective at decreasing the protein level of the siRNA target (page 57, lines 1-15; page 58, lines 11-16; and Fig. 15D and 15H).
Regarding instant claims 57 and 70, Orellana taught a conjugate comprising a receptor targeting moiety, an oligonucleotide miRNA effector molecule and an endosomal escape moiety (Fig. 1), wherein the targeting moiety, oligonucleotide miRNA effector molecule and an endosomal escape moiety are conjugated to each other with a trivalent linker
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(Fig. 3), wherein ligand-targeted delivery of the endosomal escape moiety effectively promotes cargo escape from endosomes (Fig. 5) and effectively enhances effector miRNA activity in cells (Fig. 7A-C).
Regarding instant claims 60-62 and 76, Cinci taught a method for treating the disease of cancer wherein specific delivery of CD46, CD55, and CD59 (mCRP) siRNA via the transferrin receptor (CD71) to cancer cells that expressed high levels of CD71 selectively neutralized complement regulation in cancer cells and effectively enhanced specific antibody-mediated killing of tumor cells without affecting healthy bystander cells, which is a suited strategy to improve antibody-based cancer immunotherapy (abstract).
Regarding instant claims 59-62, 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 claims 57, 65-69, and 73, 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). Regarding instant claims 57, 65-69, and 73, Weng taught SO1861 augments the escape of the genetic cargo out of the intracellular compartments into the cytosol (abstract). Regarding instant claims 57, 65-69, and 73, 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). Regarding instant claims 57, 65-69, and 73, 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). Regarding instant claims 57, 65-69, and 73, 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). Regarding instant claim 77, Weng taught an effective in vitro method for transferring an oligonucleotide effector molecule from outside a 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 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).
Regarding instant claims 71-72, King taught immunoconjugates of monoclonal antibody BR96 and a doxorubicin payload employed a hydrazone linkage to C-13 of DOX and a thioether linkage to the antibody (page 279, left column, first paragraph). King taught hydrazone linkers of this nature are stable at normal physiological pH 7, but once internalized into the acidic compartment of endosomes/lysosomes, hydrolysis occurs liberating free doxorubicin payload (page 279, left column, first paragraph). King taught BR96-DOX conjugates produced antigen-specific antitumor activity and were more potent, were more active, and produced less systemic toxicity than unconjugated DOX in vivo (page 279, left column, first paragraph). King taught the branched series of BR96 conjugates demonstrated antigen-specific cytotoxicity, and were more potent in vitro than the single-chain conjugate on both a doxorubicin and antibody basis (abstract).
Regarding instant claims 66 and 72, 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, 62-74, and 76-77, it would have been obvious for a person having ordinary skill in the art to take copending claims:
1-4, 6, 9, 11, 17, 21-23 of '760;
1, 4, 7-11, 13, 26, 32-34, and 43 of '193;
29-31, 37, 41, 45, 60, and 62 of '163
of an antibody targeted saponin and/or oligonucleotide, wherein the saponin is SO1861 – and:
i) Combine the antibody, oligonucleotide siRNA effector, and endosomal escape moiety into one conjugate as taught by Geall and include them in a pharmaceutical composition with a pharmaceutical excipient for cancer treatment in view of Geall; ii) include a trivalent linker wherein the antibody, oligonucleotide siRNA effector, and endosomal escape moiety are conjugated to each other in view of Orellana;
Include the oligonucleotide target for CD46, CD55, and CD59 (mCRP) for cancer treatment and combine the treatment with an antibody in view of Cinci;
Use a VHH CD71 targeting moiety as taught by Cohen;
Include a bivalent CD71 targeted single domain antibody, wherein the bivalent antibodies are the same in view of ‘163, ‘193, Geall, and Niewoehner;
Include in the trivalent attachment linker: i) an acid sensitive hydrazone linker to the SO1861 endosomal escape moiety at the C-23 aldehyde and an acid sensitive hydrazone linker to the effector molecule in view of King; and ii) a single or branched polymer for linking the endosomal escape moiety in view of King and Geall;
Include an oligonucleotide effector of a phosphoramidate morpholino oligomer (PMO) as taught by ‘163 and Geall; 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:
i) Geall taught a combined conjugate wherein the antibody, oligonucleotide siRNA effector, and endosomal escape moiety in one conjugate; and ii) Orellana taught a conjugate comprising a receptor targeting moiety, an oligonucleotide miRNA effector molecule and an endosomal escape moiety conjugated to each other with a trivalent linker, wherein ligand-targeted delivery of the endosomal escape moiety effectively promotes cargo escape from endosomes and effectively enhances effector miRNA activity in cells;
Cinci taught a method for treating the disease of cancer wherein specific delivery of CD46, CD55, and CD59 (mCRP) siRNA through transferrin receptor (CD71) to cancer cells that expressed high levels of CD71 selectively neutralized complement regulation in cancer cells and effectively enhanced specific antibody-mediated killing of tumor cells without affecting healthy bystander cells, which is a suited strategy to improve antibody-based cancer immunotherapy;
CD71 conjugated oligonucleotides were known to be effective in view of Cohen;
i) Geall taught the antibody as targeting CD71 with an antibody siRNA conjugate effectively and as a sdAb; ii) 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;
i) King taught immunoconjugates of monoclonal antibody BR96 and a doxorubicin payload with a single or branched polymer payload were effective in cancer cells and employed a hydrazone linkage to C-13 of DOX and a thioether linkage to the antibody, wherein hydrazone linkers of this nature are stable at normal physiological pH 7, but once internalized into the acidic compartment of endosomes/lysosomes, hydrolysis occurs liberating the free payload. 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, a hydrazone linker would release the SO1861 payload and oligonucleotide effector in the endosomes/lysosomes, where SO1861 has its effects; ii) Geall taught a polymeric molecule to which more than one endosome escape agent is bound;
Geall taught PMO as acceptable oligonucleotide effectors for antibody targeted treatment, wherein PMO are oligonucleotide effectors that target transcripts for the same purpose and would be expected to also be effective;
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) The combined conjugate would target the same cells with the antibody, oligonucleotide siRNA effector, and endosomal escape moiety in one conjugate; and ii) Orellana taught a conjugate comprising a receptor targeting moiety, an oligonucleotide miRNA effector molecule and an endosomal escape moiety conjugated to each other with a trivalent linker, wherein ligand-targeted delivery of the endosomal escape moiety effectively promotes cargo escape from endosomes and effectively enhances effector miRNA activity in cells;
Cinci taught a method for treating the disease of cancer wherein specific delivery of CD46, CD55, and CD59 (mCRP) siRNA through transferrin receptor (CD71) to cancer cells that expressed high levels of CD71 selectively neutralized complement regulation in cancer cells and effectively enhanced specific antibody-mediated killing of tumor cells without affecting healthy bystander cells, which is a suited strategy to improve antibody-based cancer immunotherapy;
CD71 conjugated oligonucleotides were known to be effective in view of Cohen;
i) Geall taught the antibody as targeting CD71 with an antibody siRNA conjugate effectively; ii) 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;
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 and effector payloads in the endosomes/lysosomes, where SO1861 has its effects;
Geall taught PMO as acceptable oligonucleotide effectors for antibody targeted treatment, wherein PMO are oligonucleotide effectors that target transcripts for the same purpose and would be expected to also be effective; 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 (instant claim 76) of a subject with a cancer overexpressing CD71, which is a cell surface receptor (instant claim 60), by silencing a protein in a cancer cell comprising administering a pharmaceutical composition comprising a pharmaceutically acceptable excipient (instant claim 75) and:
a conjugate comprising a bivalent VHH (instant claim 58) single domain CD71 targeted antibody (instant claims 61-62), a phosphoramidate morpholino oligomer (PMO) oligonucleotide effector molecule (instant claim 63) oligonucleotide targeting CD46, CD55, and CD59 (mCRP), and a single or polymeric molecule (instant claim 74) with modified SO1861 (instant claims 68-69) endosomal escape agents which has 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 65), wherein the VHH, PMO and SO1861 are covalently linked together with a trivalent linker that comprises hydrazone linkers attached to the SO1861 and PMO (instant claim 70), wherein the saponin comprises a first saccharide chain bound to the C3-beta-OH group of an aglycone core structure comprising a the glucuronic acid moiety (instant claims 67 and 73), wherein the SO1861 linker comprises an acid sensitive linker comprising a hydrazone bond linker connected at the C23 aldehyde of SO1861 (instant claims 66 and 71-72) 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 wherein the bivalent CD71 conjugate contains two VHH that are the same (instant claim 59) (instant claim 57).
Regarding instant claim 77, 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, Cohen, Orellana, Cinci, Niewoehner, Weng, King, and Marciani above of a pharmaceutical composition comprising:
a conjugate comprising a bivalent VHH single domain CD71 targeted antibody, a phosphoramidate morpholino oligomer (PMO) oligonucleotide effector molecule oligonucleotide targeting CD46, CD55, and CD59 (mCRP), and a single or polymeric molecule with modified SO1861 endosomal escape agents, wherein the VHH, PMO and SO1861 are covalently linked together with a trivalent linker that comprises hydrazone linkers attached to the SO1861 and PMO, wherein the saponin comprises a first saccharide chain bound to the C3-beta-OH group of an aglycone core structure comprising a the glucuronic acid moiety, wherein the SO1861 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 wherein the bivalent CD71 conjugate contains two VHH that are the same
– and: include the composition 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 composition comprising SO1861 and 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 conjugate comprising a bivalent VHH single domain CD71 targeted antibody, a phosphoramidate morpholino oligomer (PMO) oligonucleotide effector molecule oligonucleotide targeting CD46, CD55, and CD59 (mCRP), and a single or polymeric molecule with modified SO1861 endosomal escape agents, wherein the VHH, PMO and SO1861 are covalently linked together with a trivalent linker that comprises hydrazone linkers attached to the SO1861 and PMO, wherein the saponin comprises a first saccharide chain bound to the C3-beta-OH group of an aglycone core structure comprising a the glucuronic acid moiety, wherein the SO1861 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 wherein the bivalent CD71 conjugate contains two VHH that are the same;
providing a cell that expresses CD71 on the surface wherein the surface of the cell is targeted by the pharmaceutical compositions in a); and
contacting the cell of step b) in vitro with the pharmaceutical composition of step a),
and thus establishing the transfer of the oligonucleotide effector from outside the cell into the cell.
This is a provisional nonstatutory double patenting rejection.
Claims 57-77 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.), WO 2020144233 (Cohen R et al.), Orellana E et al. (Mol Ther Nucleic Acids. 2019 Apr 11;16:505–518. doi: 10.1016/j.omtn.2019.04.003), Cinci M et al. (Target Oncol 2015 10(3):405-413), 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, IDS reference), King HD et al. (Bioconjugate Chem. 1999, 10, 2, 279–288, IDS reference), US 2004/0242502 (Marciani D et al.), and Xian Z-Y et al. Inhibition of LDHA suppresses tumor progression in prostate cancer. (Tumour Biol. 2015 May 16;36(10):8093–8100.).
The claims of ‘760, ‘193, ‘163 and Geall, Orellana, Cinci, Niewoehner, Weng, King, and Marciani teach the limitations of instant claims 57-63 and 65-77 as set forth above.
‘760, ‘193, ‘163 and Geall, Orellana, Cinci, Niewoehner, Weng, King, and Marciani are described above.
Regarding instant claim 64, Cinci further taught a method for treating the disease of cancer wherein specific delivery of CD46, CD55, and CD59 (mCRP) siRNA through transferrin receptor (CD71) to cancer cells that expressed high levels of CD71 such as DU145 selectively neutralized complement regulation in cancer cells and effectively enhanced specific antibody-mediated killing of tumor cells without affecting healthy bystander cells, which is a suited strategy to improve antibody-based cancer immunotherapy (abstract and Figs. 1-4).
‘760, ‘193, ‘163 did not teach targeting an mRNA of LDHA, but this is obvious in view of Xian.
Xian taught decreased expression of LDHA by siRNA inhibited cell proliferation, migration, invasion, and promoted cell apoptosis of PC-3 and DU145 cells and indicated LDHA as a cancer therapeutic target (abstract and Fig. 2)
Regarding instant claim 64, it would have been obvious for a person having ordinary skill in the art to take the method of ‘760, ‘193, ‘163, Geall, Orellana, Cinci, Niewoehner, Weng, King, and Marciani above and in the pharmaceutical composition: – 1) exchange the CD46, CD55, and CD59 (mCRP) targeting effector oligonucleotide in the conjugate with an LDHA oligonucleotide conjugate of Xian.
This is obvious because: 1) Xian taught decreased expression of LDHA by siRNA inhibited cell proliferation, migration, invasion, and promoted cell apoptosis of PC-3 and DU145 cells and indicated LDHA as a cancer therapeutic target.
There is a reasonable expectation of success because: 1) Xian taught decreased expression of LDHA by siRNA inhibited cell proliferation, migration, invasion, and promoted cell apoptosis of PC-3 and DU145 cells and indicated LDHA as a cancer therapeutic target. A PMO targeting LDHA would thus be expected to block LDHA and effectively decrease cancer growth.
This is a provisional nonstatutory double patenting rejection.
Conclusion
Claims 57-77 are rejected.
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/J.J.S./Examiner, Art Unit 1643
/Karen A. Canella/Primary Examiner, Art Unit 1643
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