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
Claim status
Claims 1-6, and 17-36 are pending
Claims 19, 28-36 are withdrawn
Claims 1-6, 17-18, and 20-27 are under examination
Election/Restrictions
Applicant’s election of the following invention without traverse in the reply filed on 2/04/2026 is acknowledged.
The requirement is still deemed proper and is therefore made FINAL.
Group I, claims 1-6 and 17-27, drawn to a method of modifying an immune cell comprising a modified mRNA encoding a CAR.
Claims 28-36 are withdrawn from further consideration pursuant to 37 CFR 1.142(b) as being drawn to a nonelected invention, there being no allowable linking claim.
Applicant’s election of the following species is acknowledged.
Sunitinib as the RNAase L inhibitor.
Claim 19 is withdrawn from further consideration pursuant to 37 CFR 1.142(b) as being drawn to a nonelected species, there being no allowable generic claim.
Information Disclosure Statement
The information disclosure statements (IDS) submitted on 7/20/2023 and 2/04/2026 are in compliance with the provisions of 37 CFR 1.97. Accordingly, the information disclosure statement is being considered by the examiner.
However, Applicant is reminded that the listing of references in the specification is not a proper information disclosure statement. 37 CFR 1.98(b) requires a list of all patents, publications, or other information submitted for consideration by the Office, and MPEP § 609.04(a) states, "the list may not be incorporated into the specification but must be submitted in a separate paper." Therefore, unless the references have been cited by the examiner on form PTO-892, they have not been considered.
Claim Objections
Claim 3 is objected to because of the following informalities: instant claim uses the abbreviations “AGCap1” and “m6AGCap1”, which have not been spelled out upon first use.
Claim 22 is objected to because of the following informalities: instant claim uses the abbreviation “STNGL”, which have not been spelled out upon first use.
Although claims are allowed abbreviations, Applicant is to only use abbreviations that are well known and would be clear to someone who had not read the invention description.
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.
Claim 22 is rejected under 35 U.S.C. 112(b) or pre-AIA 35 U.S.C. 112, 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.
Where applicant acts as his or her own lexicographer to specifically define a term of a claim contrary to its ordinary meaning, the written description must clearly redefine the claim term and set forth the uncommon definition so as to put one reasonably skilled in the art on notice that the applicant intended to so redefine that claim term. Process Control Corp. v. HydReclaim Corp., 190 F.3d 1350, 1357, 52 USPQ2d 1029, 1033 (Fed. Cir. 1999). The accepted meaning for the term “cytokine” encompasses a category of small secreted proteins important in cell signaling, while instant claim uses the term to include “a recombinant 4-1BB receptor” which is not a cytokine but a co-stimulatory receptor, “beta-glucan” which is not a cytokine but is a sugar, “glucocorticoid” which is not a cytokine but is a steroid, and “immune complex”. The term “cytokine” is indefinite because the specification does not clearly redefine the term.
Claim Rejections - 35 USC § 102
The following is a quotation of the appropriate paragraphs of 35 U.S.C. 102 that form the basis for the rejections under this section made in this Office action:
A person shall be entitled to a patent unless –
(a)(1) the claimed invention was patented, described in a printed publication, or in public use, on sale or otherwise available to the public before the effective filing date of the claimed invention.
(a)(2) the claimed invention was described in a patent issued under section 151, or in an application for patent published or deemed published under section 122(b), in which the patent or application, as the case may be, names another inventor and was effectively filed before the effective filing date of the claimed invention.
Claims 1, 2, 5, 6, 21-22, and 25-27 are rejected under 35 U.S.C. 102(a)(1)/(a)(2) as being anticipated by Gill et al., (US2018/0244748, filed 7/28/2016, published 8/30/2018, see IDS filed 7/20/2023).
In regard to claim 1, Gill teaches an in vitro method of modifying human macrophages comprising the following steps:
Modifying a mRNA encoding a CAR produced by a mMessage mMachine T7 ultras In Vitro Transcription kit,
Purifying the mRNA using a Qiagen RNEasy RNA purification kit, and
Delivering the mRNA into the macrophages by electroporation ([0325], Figs 12 & 13, see also Claims 18-20 of Gill).
In regard to claim 2, Gill teaches the mRNA is modified have both a cap on the 5’end and a 3’ poly(A) tail which determine ribosome binding, initiation of translation and stability of the mRNA [0267, 0272].
In regard to claim 5, as stated supra, Gill teaches the purification step using the RNA using a Qiagen RNEasy RNA purification kit, which comprises a silica membrane purification column.
In regard to claim 6, as stated supra, Gill teaches transfection by electroporation.
In regard to claims 21, 22, and 25, Gill teaches the CAR macrophages are cultured for 30 min with beta-glucan and then tested in an in vitro killing assay [0326]
In regard to claim 25, as stated supra, Gill teaches the macrophages express the CAR.
In regard to claims 26 and 27, as stated supra, Gill teaches the mRNA is modified with a 5’cap and a 3’ poly(A) tail which contributed to stability of the mRNA. In regard to the wherein clause of claim 26, increased mRNA stability would have increased expression of the CAR relative to an unmodified mRNA. In regard to the wherein clause of claim 27, Gill teaches effector activity (as measured by phagocytosis) trends with transfection efficiency (see Fig. 15), and thus it would have been recognized that increased mRNA stability and increased expression would have increased effector activity relative to unmodified mRNA in the same cell type.
Accordingly, Gill anticipates instant claims.
Claim Rejections - 35 USC § 103
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 of this title, 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.
The factual inquiries set forth in Graham v. John Deere Co., 383 U.S. 1, 148 USPQ 459 (1966), that are applied 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.
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.
Claim 3 is rejected under 35 U.S.C. 103 as being unpatentable over Gill et al., (US2018/0244748, filed 7/28/2016, published 8/30/2018, see IDS filed 7/20/2023), in view of Grudzien-Nogalska (Meth Enzym, 2007, 431:203-227)
As discussed previously, Gill teaches an in vitro method of modifying macrophages comprising transfection with modified mRNA encoding a CAR.
However, although Gill discloses the mRNA is modified such as with a 5’ cap, they are silent to an ARAC 5’ cap.
In regard to claim 3, Grudzien-Nogalska reviews ARACs and their applications in mRNA translation and stability (p. 204-224).
Accordingly, it would have been prima facie obvious to one of ordinary skill in the art at the time of filing to practice the method of modifying macrophages comprising transfection with modified CAR mRNA comprising a 5’cap as taught by Gill and to choose the ARAC as taught by Gurdzien-Nogalska with a reasonable expectation of success. The ordinary skilled artisan would have been motivated to do so as taught by Gurdzien-Nogalska because ARACs show two-fold higher translational efficiency compared to conventional caps (p. 205, Introduction).
Hence, the claimed invention as a whole was prima facie obvious in the absence of evidence to the contrary.
Claim 4 is rejected under 35 U.S.C. 103 as being unpatentable over Gill et al., (US2018/0244748, filed 7/28/2016, published 8/30/2018, see IDS filed 7/20/2023), in view of Kariko et al. (Mol Ther, 2008, 16:1833-1840)
As discussed previously, Gill teaches a method of modifying macrophages comprising transfection with modified mRNA encoding a CAR.
However, although Gill suggest using nucleotide analogues [0265], they are silent to the mRNA being modified with a pseudouridine.
In regard to claim 4, Kariko teaches methods of expressing mRNA in cells, wherein the mRNA is modified to include psuedouridine (Abstract).
Accordingly, it would have been prima facie obvious to one of ordinary skill in the art at the time of filing to practice the method of modifying macrophages comprising transfection with modified CAR mRNA as taught by Gill and substitute modified nucleosides such as a pseudouridine as taught by Kariko with a reasonable expectation of success. The ordinary skilled artisan would have been motivated to do so as taught by Kariko because these modified mRNAs have higher translational capacity than unmodified mRNAs (Abstract).
Hence, the claimed invention as a whole was prima facie obvious in the absence of evidence to the contrary.
Claims 17 and 20 are rejected under 35 U.S.C. 103 as being unpatentable over Gill et al., (US2018/0244748, filed 7/28/2016, published 8/30/2018, see IDS filed 7/20/2023), in view of Sahin et al. (WO2014/072061, filed 11/07/2013, published 5/15/2014)
As discussed previously, Gill teaches a method of modifying macrophages comprising transfection with modified mRNA encoding CAR
However, Gill is silent with respect to further including step of treating the cells with an RNAaseL inhibitor.
Sahir teaches methods of overexpressing IVT modified RNA in cells including macrophages (Abstract, p. 67, 1st para., see Claims 1-5 of Sahir).
In regard to claim 17, Sahin teaches the steps of inhibiting intracellular IFN signaling by inhibiting RNAseL (p. 3, 4th para., p. 49, 1st para., see Claims 12-15 and 30 of Sahir).
In regard to claim 20, Sahin teaches the cells are treated with the inhibitor prior to delivering the mRNA encoding the protein to be expressed (p. 50, 4th para.).
Accordingly, it would have been prima facie obvious to one of ordinary skill in the art at the time of filing to practice the method of modifying macrophages comprising transfection with modified mRNA encoding CAR as taught by Gill and to combine step of inhibiting RNaseL as taught by Sahin with a reasonable expectation of success. The ordinary skilled artisan would have been motivated to do so as taught by Sahin because if the intracellular IFN signaling mediated by RNaseL is not inhibited, then RNA degradation occurs and/or the inhibition of RNA translation (p. 3, 4th para., p. 49, 1st para.). In regard to choosing to treat the cells with the RNaseL inhibitor before RNA delivery, it would have been obvious to choose the pretreatment option so as to avoid mRNA degradation upon delivery.
Hence, the claimed invention as a whole was prima facie obvious in the absence of evidence to the contrary.
Claim 18 is rejected under 35 U.S.C. 103 as being unpatentable over Gill et al., (US2018/0244748, filed 7/28/2016, published 8/30/2018, see IDS filed 7/20/2023), in view of Sahin et al. (WO2014/072061, filed 11/07/2013, published 5/15/2014), as applied to claim 17, in further view of Jha et al. (JBC, 2011, 286:26319-26326)
As discussed previously, Gill in view of Sahin suggest a method of modifying macrophages comprising transfection with modified mRNA encoding CAR and inhibiting RNaseL.
However, although Sahir teaches the inhibitor of IFN intracellular signaling is an imidazole-oxindole compound, Gill et al. are silent with respect the RNAaseL inhibitor as the oxindole compound of sinitunib.
Jha teaches that sunitinib is a potent inhibitor of RNaseL in cultured mammalian cells (Abstract, Introduction).
Accordingly, it would have been prima facie obvious to one of ordinary skill in the art at the time of filing to practice the method of modifying macrophages with a CAR mRNA and further combine an RNaseL inhibitor to improve mRNA expression as suggested by Gill and Sahin and choose sunitinib as the RNaseL inhibitor as taught by Jha with a reasonable expectation of success. The ordinary skilled artisan would have been motivated to do so as taught by Jha because of the in vitro potency of sunitinib in cultured mammalian cells.
Hence, the claimed invention as a whole was prima facie obvious in the absence of evidence to the contrary.
Claims 21-24 are rejected under 35 U.S.C. 103 as being unpatentable over Gill et al., (US2018/0244748, filed 7/28/2016, published 8/30/2018, see IDS filed 7/20/2023), in view of Xie et al. (IJMM, 2016, 38:148-160)
As discussed previously, Gill teaches a method of modifying macrophages comprising transfection with modified mRNA encoding CAR.
However, although Gill suggests the method steps of stimulating the CAR monocytes (i.e., CARMA) by treatments with synergistic compounds such as cytokines after delivering of the CAR nucleic acid ([0127, 0384], Fig. 28), and Gill discloses the ability of macrophages to become polarized, and teaches the anti-tumor advantages of up-regulation of M1 markers in inflammatory/activated macrophages [0012, 0283, 0374], see also Claims 1 and 9 of Gill), Gill does not reduce to practice the step of culturing the RNA transfected macrophages with IFN-beta.
Xie teaches methods of treating macrophages with cytokines in order to induce M1 polarization and teaches that media comprising IFN-beta is required for sustained M1 polarization (Abstract a chimeric receptors (Abstract).
Accordingly, it would have been prima facie obvious to one of ordinary skill in the art at the time of filing to practice the method of modifying macrophages comprising transfection with modified mRNA encoding CAR and to treat the CARMA with cytokines to induce a M1 phenotype as suggested by Gill and to combine IFN-beta in the cytokine treatment as suggested by Xie with a reasonable expectation of success. The ordinary skilled artisan would have been motivated to do so as taught by Xie because IFN-beta plays a crucial role in the regulation of M1 polarization of macrophages (Abstract, Discussion, p. 158).
Hence, the claimed invention as a whole was prima facie obvious in the absence of evidence to the contrary.
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 obviousness-type 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); and 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 a nonstatutory double patenting ground provided the conflicting application or patent either is shown to be commonly owned with this application, or claims an invention made as a result of activities undertaken within the scope of a joint research agreement.
Effective January 1, 1994, a registered attorney or agent of record may sign a terminal disclaimer. A terminal disclaimer signed by the assignee must fully comply with 37 CFR 3.73(b).
Claims 1, 2, 5, 6, 21-22, and 25-27 are rejected on the grounds of nonstatutory double patenting over claims 1-10 of U.S. Patent No. 11,739,297 (Klichinsky et al., Patented 8/29/2023), in view of Gill et al., (US2018/0244748, filed 7/28/2016, published 8/30/2018, see IDS filed 7/20/2023).
The subject matter claimed in the instant application is fully disclosed in the referenced patent as follows: the method for modifying a macrophage or monocyte comprising delivering a nucleic acid that encodes a CAR of cited patent makes obvious the method of instant application. It is clear that all the elements of the cited patent claims are to be found in instant claims. The difference between the cited patent claims and the instant claims lies in the fact that the instant claims recite that the nucleic acid is a purified modified mRNA.
In regard to claim 1, Gill teaches an in vitro method of modifying human macrophages comprising the following steps:
Modifying a mRNA encoding a CAR produced by a mMessage mMachine T7 ultras In Vitro Transcription kit,
Purifying the mRNA using a Qiagen RNEasy RNA purification kit, and
Delivering the mRNA into the macrophages by electroporation ([0325], Figs 12 & 13, see also Claims 18-20 of Gill).
In regard to claim 2, Gill teaches the mRNA is modified have both a cap on the 5’end and a 3’ poly(A) tail which determine ribosome binding, initiation of translation and stability of the mRNA [0267, 0272].
Accordingly it would have been obvious to claim the cited method and to further claim a purified modified mRNA as taught by Gill with a reasonable expectation of success. One of ordinary skill in the art would have claimed so because of the high efficiency of CAR transfection with purified modified RNA.
In regard to claim 5, as stated supra, Gill teaches the purification step using the RNA using a Qiagen RNEasy RNA purification kit, which comprises a silica membrane purification column.
In regard to claim 6, as stated supra, Gill teaches transfection by electroporation in order to achieve high efficiency transfection.
In regard to claims 21, 22, and 25, Gill teaches the CAR macrophages are cultured for 30 min with beta-glucan and then tested in an in vitro killing assay [0326], which would have been obvious to claim so as to test the efficacy of the CARMA.
In regard to claim 25, as stated supra, Gill teaches the macrophages express the CAR.
In regard to claims 26 and 27, as stated supra, Gill teaches the mRNA is modified with a 5’cap and a 3’ poly(A) tail which contributed to stability of the mRNA.
Since the instant application claims are obvious over cited patent claims in view of Gill, said claims are not patentably distinct.
Claim 3 is rejected on the grounds of nonstatutory double patenting over claims 1-10 of U.S. Patent No. 11,739,297 (Klichinsky et al., Patented 8/29/2023), in view of Gill et al., (US2018/0244748, filed 7/28/2016, published 8/30/2018, see IDS filed 7/20/2023), in view further of Grudzien-Nogalska (Meth Enzym, 2007, 431:203-227)
As discussed previously, Gill makes obvious an in vitro method of modifying macrophages comprising transfection with modified mRNA encoding a CAR.
However, although Gill makes obvious the mRNA is modified such as with a 5’ cap, they are silent to an ARAC 5’ cap.
In regard to claim 3, Grudzien-Nogalska reviews ARACs and their applications in mRNA translation and stability (p. 204-224).
Accordingly, it would have been prima facie obvious to one of ordinary skill in the art at the time of filing to claim the method of modifying macrophages comprising transfection with modified CAR mRNA comprising a 5’cap as taught by Gill and to choose the ARAC as taught by Gurdzien-Nogalska with a reasonable expectation of success. The ordinary skilled artisan would have been motivated to do so as taught by Gurdzien-Nogalska because ARACs show two-fold higher translational efficiency compared to conventional caps (p. 205, Introduction).
Since the instant application claims are obvious over cited patent claims in view of Gill et al., said claims are not patentably distinct.
Claim 4 is rejected on the grounds of nonstatutory double patenting over claims 1-10 of U.S. Patent No. 11,739,297 (Klichinsky et al., Patented 8/29/2023), in view of Gill et al., (US2018/0244748, filed 7/28/2016, published 8/30/2018, see IDS filed 7/20/2023), in further view of Kariko et al. (Mol Ther, 2008, 16:1833-1840)
As discussed previously, Gill makes a method of modifying macrophages comprising transfection with modified mRNA encoding a CAR.
However, although Gill suggest using nucleotide analogues [0265], they are silent to the mRNA being modified with a pseudouridine.
In regard to claim 4, Kariko teaches methods of expressing mRNA in cells, wherein the mRNA is modified to include psuedouridine (Abstract).
Accordingly, it would have been prima facie obvious to one of ordinary skill in the art at the time of filing to claim the method of modifying macrophages comprising transfection with modified CAR mRNA as taught by Gill and substitute modified nucleosides such as a pseudouridine as taught by Kariko with a reasonable expectation of success. The ordinary skilled artisan would have been motivated to do so as taught by Kariko because these modified mRNAs have higher translational capacity than unmodified mRNAs (Abstract).
Since the instant application claims are obvious over cited patent claims in view of Gill et al., said claims are not patentably distinct.
Claims 17 and 20 are rejected on the grounds of nonstatutory double patenting over claims 1-10 of U.S. Patent No. 11,739,297 (Klichinsky et al., Patented 8/29/2023), in view of Gill et al., (US2018/0244748, filed 7/28/2016, published 8/30/2018, see IDS filed 7/20/2023), in further view of Sahin et al. (WO2014/072061, filed 11/07/2013, published 5/15/2014)
As discussed previously, Gill makes obvious a method of modifying macrophages comprising transfection with modified mRNA encoding CAR
However, Gill is silent with respect to further including step of treating the cells with an RNAaseL inhibitor.
Sahir teaches methods of overexpressing IVT modified RNA in cells including macrophages (Abstract, p. 67, 1st para., see Claims 1-5 of Sahir).
In regard to claim 17, Sahin teaches the steps of inhibiting intracellular IFN signaling by inhibiting RNAseL (p. 3, 4th para., p. 49, 1st para., see Claims 12-15 and 30 of Sahir).
In regard to claim 20, Sahin teaches the cells are treated with the inhibitor prior to delivering the mRNA encoding the protein to be expressed (p. 50, 4th para.).
Accordingly, it would have been prima facie obvious to one of ordinary skill in the art at the time of filing to claim the method of modifying macrophages comprising transfection with modified mRNA encoding CAR as taught by Gill and to combine step of inhibiting RNaseL as taught by Sahin with a reasonable expectation of success. The ordinary skilled artisan would have been motivated to do so as taught by Sahin because if the intracellular IFN signaling mediated by RNaseL is not inhibited, then RNA degradation occurs and/or the inhibition of RNA translation (p. 3, 4th para., p. 49, 1st para.). In regard to choosing to treat the cells with the RNaseL inhibitor before RNA delivery, it would have been obvious to choose the pretreatment option so as to avoid mRNA degradation upon delivery.
Since the instant application claims are obvious over cited patent claims in view of Gill et al., said claims are not patentably distinct.
Claim 18 is rejected are rejected on the grounds of nonstatutory double patenting over claims 1-10 of U.S. Patent No. 11,739,297 (Klichinsky et al., Patented 8/29/2023), in view of Gill et al., (US2018/0244748, filed 7/28/2016, published 8/30/2018, see IDS filed 7/20/2023) and Sahin et al. (WO2014/072061, filed 11/07/2013, published 5/15/2014, in further view of Jha et al. (JBC, 2011, 286:26319-26326)
As discussed previously, Gill in view of Sahin make obvious a method of modifying macrophages comprising transfection with modified mRNA encoding CAR and inhibiting RNaseL.
However, although Sahir teaches the inhibitor of IFN intracellular signaling is an imidazole-oxindole compound, Gill et al. are silent with respect the RNAaseL inhibitor as the oxindole compound of sinitunib.
Jha teaches that sunitinib is a potent inhibitor of RNaseL in cultured mammalian cells (Abstract, Introduction).
Accordingly, it would have been prima facie obvious to one of ordinary skill in the art at the time of filing to claim the method of modifying macrophages with a CAR mRNA and further combine an RNaseL inhibitor to improve mRNA expression as suggested by Gill and Sahin and choose sunitinib as the RNaseL inhibitor as taught by Jha with a reasonable expectation of success. The ordinary skilled artisan would have been motivated to do so as taught by Jha because of the in vitro potency of sunitinib in cultured mammalian cells.
Since the instant application claims are obvious over cited patent claims in view of Gill et al., said claims are not patentably distinct.
Claims 21-24 are rejected on the grounds of nonstatutory double patenting over claims 1-10 of U.S. Patent No. 11,739,297 (Klichinsky et al., Patented 8/29/2023), in view of Gill et al., (US2018/0244748, filed 7/28/2016, published 8/30/2018, see IDS filed 7/20/2023), in view of Xie et al. (IJMM, 2016, 38:148-160)
As discussed previously, Gill make obvious a method of modifying macrophages comprising transfection with modified mRNA encoding CAR.
However, although Gill suggests the method steps of stimulating the CAR monocytes (i.e., CARMA) by treatments with synergistic compounds such as cytokines after delivering of the CAR nucleic acid ([0127, 0384], Fig. 28), and Gill discloses the ability of macrophages to become polarized, and teaches the anti-tumor advantages of up-regulation of M1 markers in inflammatory/activated macrophages [0012, 0283, 0374], see also Claims 1 and 9 of Gill), Gill does not reduce to practice the step of culturing the RNA transfected macrophages with IFN-beta.
Xie teaches methods of treating macrophages with cytokines in order to induce M1 polarization and teaches that media comprising IFN-beta is required for sustained M1 polarization (Abstract a chimeric receptors (Abstract).
Accordingly, it would have been prima facie obvious to one of ordinary skill in the art at the time of filing to claim the method of modifying macrophages comprising transfection with modified mRNA encoding CAR and to treat the CARMA with cytokines to induce a M1 phenotype as suggested by Gill and to combine IFN-beta in the cytokine treatment as suggested by Xie with a reasonable expectation of success. The ordinary skilled artisan would have been motivated to do so as taught by Xie because IFN-beta plays a crucial role in the regulation of M1 polarization of macrophages (Abstract, Discussion, p. 158).
Since the instant application claims are obvious over cited patent claims in view of Gill et al., said claims are not patentably distinct.
Claims 1, 2, 5, 6, 21-22, and 25-27 are rejected on the grounds of nonstatutory double patenting over claims 17-24 of U.S. Patent No. 11,498,954 (Gill, Klichinsky, and June, Patented11/15/2022), in view of Gill et al., (US2018/0244748, filed 7/28/2016, published 8/30/2018, see IDS filed 7/20/2023).
The subject matter claimed in the instant application is fully disclosed in the referenced patent as follows: the method for modifying a macrophage or monocyte comprising delivering a nucleic acid that encodes a CAR of cited patent makes obvious the method of instant application. It is clear that all the elements of the cited patent claims are to be found in instant claims. The difference between the cited patent claims and the instant claims lies in the fact that the instant claims recite that the nucleic acid is a purified modified mRNA.
In regard to claim 1, Gill teaches an in vitro method of modifying human macrophages comprising the following steps:
Modifying a mRNA encoding a CAR produced by a mMessage mMachine T7 ultras In Vitro Transcription kit,
Purifying the mRNA using a Qiagen RNEasy RNA purification kit, and
Delivering the mRNA into the macrophages by electroporation ([0325], Figs 12 & 13, see also Claims 18-20 of Gill).
In regard to claim 2, Gill teaches the mRNA is modified have both a cap on the 5’end and a 3’ poly(A) tail which determine ribosome binding, initiation of translation and stability of the mRNA [0267, 0272].
Accordingly it would have been obvious to claim the cited method and to further claim a purified modified mRNA as taught by Gill with a reasonable expectation of success. One of ordinary skill in the art would have claimed so because of the high efficiency of CAR transfection with purified modified RNA.
In regard to claim 5, as stated supra, Gill teaches the purification step using the RNA using a Qiagen RNEasy RNA purification kit, which comprises a silica membrane purification column.
In regard to claim 6, as stated supra, Gill teaches transfection by electroporation in order to achieve high efficiency transfection.
In regard to claims 21, 22, and 25, Gill teaches the CAR macrophages are cultured for 30 min with beta-glucan and then tested in an in vitro killing assay [0326], which would have been obvious to claim so as to test the efficacy of the CARMA.
In regard to claim 25, as stated supra, Gill teaches the macrophages express the CAR.
In regard to claims 26 and 27, as stated supra, Gill teaches the mRNA is modified with a 5’cap and a 3’ poly(A) tail which contributed to stability of the mRNA.
Since the instant application claims are obvious over cited patent claims in view of Gill, said claims are not patentably distinct.
Claim 3 is rejected on the grounds of nonstatutory double patenting over claims 17-24 of U.S. Patent No. 11,498,954 (Gill, Klichinsky, and June, Patented11/15/2022), in view of Gill et al., (US2018/0244748, filed 7/28/2016, published 8/30/2018, see IDS filed 7/20/2023), in view further of Grudzien-Nogalska (Meth Enzym, 2007, 431:203-227)
As discussed previously, Gill makes obvious an in vitro method of modifying macrophages comprising transfection with modified mRNA encoding a CAR.
However, although Gill makes obvious the mRNA is modified such as with a 5’ cap, they are silent to an ARAC 5’ cap.
In regard to claim 3, Grudzien-Nogalska reviews ARACs and their applications in mRNA translation and stability (p. 204-224).
Accordingly, it would have been prima facie obvious to one of ordinary skill in the art at the time of filing to claim the method of modifying macrophages comprising transfection with modified CAR mRNA comprising a 5’cap as taught by Gill and to choose the ARAC as taught by Gurdzien-Nogalska with a reasonable expectation of success. The ordinary skilled artisan would have been motivated to do so as taught by Gurdzien-Nogalska because ARACs show two-fold higher translational efficiency compared to conventional caps (p. 205, Introduction).
Since the instant application claims are obvious over cited patent claims in view of Gill et al., said claims are not patentably distinct.
Claim 4 is rejected on the grounds of nonstatutory double patenting over claims 17-24 of U.S. Patent No. 11,498,954 (Gill, Klichinsky, and June, Patented11/15/2022), in view of Gill et al., (US2018/0244748, filed 7/28/2016, published 8/30/2018, see IDS filed 7/20/2023), in further view of Kariko et al. (Mol Ther, 2008, 16:1833-1840)
As discussed previously, Gill makes a method of modifying macrophages comprising transfection with modified mRNA encoding a CAR.
However, although Gill suggest using nucleotide analogues [0265], they are silent to the mRNA being modified with a pseudouridine.
In regard to claim 4, Kariko teaches methods of expressing mRNA in cells, wherein the mRNA is modified to include psuedouridine (Abstract).
Accordingly, it would have been prima facie obvious to one of ordinary skill in the art at the time of filing to claim the method of modifying macrophages comprising transfection with modified CAR mRNA as taught by Gill and substitute modified nucleosides such as a pseudouridine as taught by Kariko with a reasonable expectation of success. The ordinary skilled artisan would have been motivated to do so as taught by Kariko because these modified mRNAs have higher translational capacity than unmodified mRNAs (Abstract).
Since the instant application claims are obvious over cited patent claims in view of Gill et al., said claims are not patentably distinct.
Claims 17 and 20 are rejected on the grounds of nonstatutory double patenting over claims 17-24 of U.S. Patent No. 11,498,954 (Gill, Klichinsky, and June, Patented11/15/2022), in view of Gill et al., (US2018/0244748, filed 7/28/2016, published 8/30/2018, see IDS filed 7/20/2023), in further view of Sahin et al. (WO2014/072061, filed 11/07/2013, published 5/15/2014)
As discussed previously, Gill makes obvious a method of modifying macrophages comprising transfection with modified mRNA encoding CAR
However, Gill is silent with respect to further including step of treating the cells with an RNAaseL inhibitor.
Sahir teaches methods of overexpressing IVT modified RNA in cells including macrophages (Abstract, p. 67, 1st para., see Claims 1-5 of Sahir).
In regard to claim 17, Sahin teaches the steps of inhibiting intracellular IFN signaling by inhibiting RNAseL (p. 3, 4th para., p. 49, 1st para., see Claims 12-15 and 30 of Sahir).
In regard to claim 20, Sahin teaches the cells are treated with the inhibitor prior to delivering the mRNA encoding the protein to be expressed (p. 50, 4th para.).
Accordingly, it would have been prima facie obvious to one of ordinary skill in the art at the time of filing to claim the method of modifying macrophages comprising transfection with modified mRNA encoding CAR as taught by Gill and to combine step of inhibiting RNaseL as taught by Sahin with a reasonable expectation of success. The ordinary skilled artisan would have been motivated to do so as taught by Sahin because if the intracellular IFN signaling mediated by RNaseL is not inhibited, then RNA degradation occurs and/or the inhibition of RNA translation (p. 3, 4th para., p. 49, 1st para.). In regard to choosing to treat the cells with the RNaseL inhibitor before RNA delivery, it would have been obvious to choose the pretreatment option so as to avoid mRNA degradation upon delivery.
Since the instant application claims are obvious over cited patent claims in view of Gill et al., said claims are not patentably distinct.
Claim 18 is rejected are rejected on the grounds of nonstatutory double patenting over claims 17-24 of U.S. Patent No. 11,498,954 (Gill, Klichinsky, and June, Patented11/15/2022), in view of Gill et al., (US2018/0244748, filed 7/28/2016, published 8/30/2018, see IDS filed 7/20/2023) and Sahin et al. (WO2014/072061, filed 11/07/2013, published 5/15/2014, in further view of Jha et al. (JBC, 2011, 286:26319-26326)
As discussed previously, Gill in view of Sahin make obvious a method of modifying macrophages comprising transfection with modified mRNA encoding CAR and inhibiting RNaseL.
However, although Sahir teaches the inhibitor of IFN intracellular signaling is an imidazole-oxindole compound, Gill et al. are silent with respect the RNAaseL inhibitor as the oxindole compound of sinitunib.
Jha teaches that sunitinib is a potent inhibitor of RNaseL in cultured mammalian cells (Abstract, Introduction).
Accordingly, it would have been prima facie obvious to one of ordinary skill in the art at the time of filing to claim the method of modifying macrophages with a CAR mRNA and further combine an RNaseL inhibitor to improve mRNA expression as suggested by Gill and Sahin and choose sunitinib as the RNaseL inhibitor as taught by Jha with a reasonable expectation of success. The ordinary skilled artisan would have been motivated to do so as taught by Jha because of the in vitro potency of sunitinib in cultured mammalian cells.
Since the instant application claims are obvious over cited patent claims in view of Gill et al., said claims are not patentably distinct.
Claims 21-24 are rejected on the grounds of nonstatutory double patenting over claims 17-24 of U.S. Patent No. 11,498,954 (Gill, Klichinsky, and June, Patented11/15/2022)), in view of Gill et al., (US2018/0244748, filed 7/28/2016, published 8/30/2018, see IDS filed 7/20/2023), in view of Xie et al. (IJMM, 2016, 38:148-160)
As discussed previously, Gill make obvious a method of modifying macrophages comprising transfection with modified mRNA encoding CAR.
However, although Gill suggests the method steps of stimulating the CAR monocytes (i.e., CARMA) by treatments with synergistic compounds such as cytokines after delivering of the CAR nucleic acid ([0127, 0384], Fig. 28), and Gill discloses the ability of macrophages to become polarized, and teaches the anti-tumor advantages of up-regulation of M1 markers in inflammatory/activated macrophages [0012, 0283, 0374], see also Claims 1 and 9 of Gill), Gill does not reduce to practice the step of culturing the RNA transfected macrophages with IFN-beta.
Xie teaches methods of treating macrophages with cytokines in order to induce M1 polarization and teaches that media comprising IFN-beta is required for sustained M1 polarization (Abstract a chimeric receptors (Abstract).
Accordingly, it would have been prima facie obvious to one of ordinary skill in the art at the time of filing to claim the method of modifying macrophages comprising transfection with modified mRNA encoding CAR and to treat the CARMA with cytokines to induce a M1 phenotype as suggested by Gill and to combine IFN-beta in the cytokine treatment as suggested by Xie with a reasonable expectation of success. The ordinary skilled artisan would have been motivated to do so as taught by Xie because IFN-beta plays a crucial role in the regulation of M1 polarization of macrophages (Abstract, Discussion, p. 158).
Since the instant application claims are obvious over cited patent claims in view of Gill et al., said claims are not patentably distinct.
Claims 1, 2, 5, 6, 21-22, and 25-27 are rejected on the grounds of nonstatutory double patenting over claims 25-27 of U.S. Patent No. 11,407,805 (Gill, Klichinsky, and June, Patented 8/09/2022) in view of Gill et al., (US2018/0244748, filed 7/28/2016, published 8/30/2018, see IDS filed 7/20/2023).
The subject matter claimed in the instant application is fully disclosed in the referenced patent as follows: the method for modifying a macrophage or monocyte comprising electroporating a mRNA that encodes a CAR of cited patent makes obvious the method of instant application. It is clear that all the elements of the cited patent claims are to be found in instant claims. The difference between the cited patent claims and the instant claims lies in the fact that the instant claims recite that the mRNA is a purified modified mRNA.
In regard to claim 1, Gill teaches an in vitro method of modifying human macrophages comprising the following steps:
Modifying a mRNA encoding a CAR produced by a mMessage mMachine T7 ultras In Vitro Transcription kit,
Purifying the mRNA using a Qiagen RNEasy RNA purification kit, and
Delivering the mRNA into the macrophages by electroporation ([0325], Figs 12 & 13, see also Claims 18-20 of Gill).
In regard to claim 2, Gill teaches the mRNA is modified have both a cap on the 5’end and a 3’ poly(A) tail which determine ribosome binding, initiation of translation and stability of the mRNA [0267, 0272].
Accordingly it would have been obvious to claim the cited method and to further claim a purified modified mRNA as taught by Gill with a reasonable expectation of success. One of ordinary skill in the art would have claimed so because of the high efficiency of CAR transfection with purified modified RNA.
In regard to claim 5, as stated supra, Gill teaches the purification step using the RNA using a Qiagen RNEasy RNA purification kit, which comprises a silica membrane purification column.
In regard to claim 6, as stated supra, Gill teaches transfection by electroporation in order to achieve high efficiency transfection.
In regard to claims 21, 22, and 25, Gill teaches the CAR macrophages are cultured for 30 min with beta-glucan and then tested in an in vitro killing assay [0326], which would have been obvious to claim so as to test the efficacy of the CARMA.
In regard to claim 25, as stated supra, Gill teaches the macrophages express the CAR.
In regard to claims 26 and 27, as stated supra, Gill teaches the mRNA is modified with a 5’cap and a 3’ poly(A) tail which contributed to stability of the mRNA.
Since the instant application claims are obvious over cited patent claims in view of Gill, said claims are not patentably distinct.
Claim 3 is rejected on the grounds of nonstatutory double patenting over claims 25-27 of U.S. Patent No. 11,407,805 (Gill, Klichinsky, and June, Patented 8/09/2022)), in view of Gill et al., (US2018/0244748, filed 7/28/2016, published 8/30/2018, see IDS filed 7/20/2023), in view further of Grudzien-Nogalska (Meth Enzym, 2007, 431:203-227)
As discussed previously, Gill makes obvious an in vitro method of modifying macrophages comprising transfection with modified mRNA encoding a CAR.
However, although Gill makes obvious the mRNA is modified such as with a 5’ cap, they are silent to an ARAC 5’ cap.
In regard to claim 3, Grudzien-Nogalska reviews ARACs and their applications in mRNA translation and stability (p. 204-224).
Accordingly, it would have been prima facie obvious to one of ordinary skill in the art at the time of filing to claim the method of modifying macrophages comprising transfection with modified CAR mRNA comprising a 5’cap as taught by Gill and to choose the ARAC as taught by Gurdzien-Nogalska with a reasonable expectation of success. The ordinary skilled artisan would have been motivated to do so as taught by Gurdzien-Nogalska because ARACs show two-fold higher translational efficiency compared to conventional caps (p. 205, Introduction).
Since the instant application claims are obvious over cited patent claims in view of Gill et al., said claims are not patentably distinct.
Claim 4 is rejected on the grounds of nonstatutory double patenting over claims 25-27 of U.S. Patent No. 11,407,805 (Gill, Klichinsky, and June, Patented 8/09/2022)), in view of Gill et al., (US2018/0244748, filed 7/28/2016, published 8/30/2018, see IDS filed 7/20/2023), in further view of Kariko et al. (Mol Ther, 2008, 16:1833-1840)
As discussed previously, Gill makes a method of modifying macrophages comprising transfection with modified mRNA encoding a CAR.
However, although Gill suggest using nucleotide analogues [0265], they are silent to the mRNA being modified with a pseudouridine.
In regard to claim 4, Kariko teaches methods of expressing mRNA in cells, wherein the mRNA is modified to include psuedouridine (Abstract).
Accordingly, it would have been prima facie obvious to one of ordinary skill in the art at the time of filing to claim the method of modifying macrophages comprising transfection with modified CAR mRNA as taught by Gill and substitute modified nucleosides such as a pseudouridine as taught by Kariko with a reasonable expectation of success. The ordinary skilled artisan would have been motivated to do so as taught by Kariko because these modified mRNAs have higher translational capacity than unmodified mRNAs (Abstract).
Since the instant application claims are obvious over cited patent claims in view of Gill et al., said claims are not patentably distinct.
Claims 17 and 20 are rejected on the grounds of nonstatutory double patenting over claims 25-27 of U.S. Patent No. 11,407,805 (Gill, Klichinsky, and June, Patented 8/09/2022)), in view of Gill et al., (US2018/0244748, filed 7/28/2016, published 8/30/2018, see IDS filed 7/20/2023), in further view of Sahin et al. (WO2014/072061, filed 11/07/2013, published 5/15/2014)
As discussed previously, Gill makes obvious a method of modifying macrophages comprising transfection with modified mRNA encoding CAR
However, Gill is silent with respect to further including step of treating the cells with an RNAaseL inhibitor.
Sahir teaches methods of overexpressing IVT modified RNA in cells including macrophages (Abstract, p. 67, 1st para., see Claims 1-5 of Sahir).
In regard to claim 17, Sahin teaches the steps of inhibiting intracellular IFN signaling by inhibiting RNAseL (p. 3, 4th para., p. 49, 1st para., see Claims 12-15 and 30 of Sahir).
In regard to claim 20, Sahin teaches the cells are treated with the inhibitor prior to delivering the mRNA encoding the protein to be expressed (p. 50, 4th para.).
Accordingly, it would have been prima facie obvious to one of ordinary skill in the art at the time of filing to claim the method of modifying macrophages comprising transfection with modified mRNA encoding CAR as taught by Gill and to combine step of inhibiting RNaseL as taught by Sahin with a reasonable expectation of success. The ordinary skilled artisan would have been motivated to do so as taught by Sahin because if the intracellular IFN signaling mediated by RNaseL is not inhibited, then RNA degradation occurs and/or the inhibition of RNA translation (p. 3, 4th para., p. 49, 1st para.). In regard to choosing to treat the cells with the RNaseL inhibitor before RNA delivery, it would have been obvious to choose the pretreatment option so as to avoid mRNA degradation upon delivery.
Since the instant application claims are obvious over cited patent claims in view of Gill et al., said claims are not patentably distinct.
Claim 18 is rejected are rejected on the grounds of nonstatutory double patenting over claims 25-27 of U.S. Patent No. 11,407,805 (Gill, Klichinsky, and June, Patented 8/09/2022), in view of Gill et al., (US2018/0244748, filed 7/28/2016, published 8/30/2018, see IDS filed 7/20/2023) and Sahin et al. (WO2014/072061, filed 11/07/2013, published 5/15/2014, in further view of Jha et al. (JBC, 2011, 286:26319-26326)
As discussed previously, Gill in view of Sahin make obvious a method of modifying macrophages comprising transfection with modified mRNA encoding CAR and inhibiting RNaseL.
However, although Sahir teaches the inhibitor of IFN intracellular signaling is an imidazole-oxindole compound, Gill et al. are silent with respect the RNAaseL inhibitor as the oxindole compound of sinitunib.
Jha teaches that sunitinib is a potent inhibitor of RNaseL in cultured mammalian cells (Abstract, Introduction).
Accordingly, it would have been prima facie obvious to one of ordinary skill in the art at the time of filing to claim the method of modifying macrophages with a CAR mRNA and further combine an RNaseL inhibitor to improve mRNA expression as suggested by Gill and Sahin and choose sunitinib as the RNaseL inhibitor as taught by Jha with a reasonable expectation of success. The ordinary skilled artisan would have been motivated to do so as taught by Jha because of the in vitro potency of sunitinib in cultured mammalian cells.
Since the instant application claims are obvious over cited patent claims in view of Gill et al., said claims are not patentably distinct.
Claims 21-24 are rejected on the grounds of nonstatutory double patenting over claims 25-27 of U.S. Patent No. 11,407,805 (Gill, Klichinsky, and June, Patented 8/09/2022), in view of Gill et al., (US2018/0244748, filed 7/28/2016, published 8/30/2018, see IDS filed 7/20/2023), in view of Xie et al. (IJMM, 2016, 38:148-160)
As discussed previously, Gill make obvious a method of modifying macrophages comprising transfection with modified mRNA encoding CAR.
However, although Gill suggests the method steps of stimulating the CAR monocytes (i.e., CARMA) by treatments with synergistic compounds such as cytokines after delivering of the CAR nucleic acid ([0127, 0384], Fig. 28), and Gill discloses the ability of macrophages to become polarized, and teaches the anti-tumor advantages of up-regulation of M1 markers in inflammatory/activated macrophages [0012, 0283, 0374], see also Claims 1 and 9 of Gill), Gill does not reduce to practice the step of culturing the RNA transfected macrophages with IFN-beta.
Xie teaches methods of treating macrophages with cytokines in order to induce M1 polarization and teaches that media comprising IFN-beta is required for sustained M1 polarization (Abstract a chimeric receptors (Abstract).
Accordingly, it would have been prima facie obvious to one of ordinary skill in the art at the time of filing to claim the method of modifying macrophages comprising transfection with modified mRNA encoding CAR and to treat the CARMA with cytokines to induce a M1 phenotype as suggested by Gill and to combine IFN-beta in the cytokine treatment as suggested by Xie with a reasonable expectation of success. The ordinary skilled artisan would have been motivated to do so as taught by Xie because IFN-beta plays a crucial role in the regulation of M1 polarization of macrophages (Abstract, Discussion, p. 158).
Since the instant application claims are obvious over cited patent claims in view of Gill et al., said claims are not patentably distinct.
Claims 1, 2, 5, 6, 21-22, and 25-27 are rejected on the grounds of nonstatutory double patenting over claims 17-19 of U.S. Patent No. 11,332,511 (Gill, Klichinsky, and June, Patented 5/17/2022) in view of Gill et al., (US2018/0244748, filed 7/28/2016, published 8/30/2018, see IDS filed 7/20/2023).
The subject matter claimed in the instant application is fully disclosed in the referenced patent as follows: the method for modifying a macrophage or monocyte comprising delivering a mRNA that encodes a CAR of cited patent makes obvious the method of instant application. It is clear that all the elements of the cited patent claims are to be found in instant claims. The difference between the cited patent claims and the instant claims lies in the fact that the instant claims recite that the mRNA acid is a purified modified mRNA.
In regard to claim 1, Gill teaches an in vitro method of modifying human macrophages comprising the following steps:
Modifying a mRNA encoding a CAR produced by a mMessage mMachine T7 ultras In Vitro Transcription kit,
Purifying the mRNA using a Qiagen RNEasy RNA purification kit, and
Delivering the mRNA into the macrophages by electroporation ([0325], Figs 12 & 13, see also Claims 18-20 of Gill).
In regard to claim 2, Gill teaches the mRNA is modified have both a cap on the 5’end and a 3’ poly(A) tail which determine ribosome binding, initiation of translation and stability of the mRNA [0267, 0272].
Accordingly it would have been obvious to claim the cited method and to further claim a purified modified mRNA as taught by Gill with a reasonable expectation of success. One of ordinary skill in the art would have claimed so because of the high efficiency of CAR transfection with purified modified RNA.
In regard to claim 5, as stated supra, Gill teaches the purification step using the RNA using a Qiagen RNEasy RNA purification kit, which comprises a silica membrane purification column.
In regard to claim 6, as stated supra, Gill teaches transfection by electroporation in order to achieve high efficiency transfection.
In regard to claims 21, 22, and 25, Gill teaches the CAR macrophages are cultured for 30 min with beta-glucan and then tested in an in vitro killing assay [0326], which would have been obvious to claim so as to test the efficacy of the CARMA.
In regard to claim 25, as stated supra, Gill teaches the macrophages express the CAR.
In regard to claims 26 and 27, as stated supra, Gill teaches the mRNA is modified with a 5’cap and a 3’ poly(A) tail which contributed to stability of the mRNA.
Since the instant application claims are obvious over cited patent claims in view of Gill, said claims are not patentably distinct.
Claim 3 is rejected on the grounds of nonstatutory double patenting over 17-19 of U.S. Patent No. 11,332,511 (Gill, Klichinsky, and June, Patented 5/17/2022)), in view of Gill et al., (US2018/0244748, filed 7/28/2016, published 8/30/2018, see IDS filed 7/20/2023), in view further of Grudzien-Nogalska (Meth Enzym, 2007, 431:203-227)
As discussed previously, Gill makes obvious an in vitro method of modifying macrophages comprising transfection with modified mRNA encoding a CAR.
However, although Gill makes obvious the mRNA is modified such as with a 5’ cap, they are silent to an ARAC 5’ cap.
In regard to claim 3, Grudzien-Nogalska reviews ARACs and their applications in mRNA translation and stability (p. 204-224).
Accordingly, it would have been prima facie obvious to one of ordinary skill in the art at the time of filing to claim the method of modifying macrophages comprising transfection with modified CAR mRNA comprising a 5’cap as taught by Gill and to choose the ARAC as taught by Gurdzien-Nogalska with a reasonable expectation of success. The ordinary skilled artisan would have been motivated to do so as taught by Gurdzien-Nogalska because ARACs show two-fold higher translational efficiency compared to conventional caps (p. 205, Introduction).
Since the instant application claims are obvious over cited patent claims in view of Gill et al., said claims are not patentably distinct.
Claim 4 is rejected on the grounds of nonstatutory double patenting over 17-19 of U.S. Patent No. 11,332,511 (Gill, Klichinsky, and June, Patented 5/17/2022)), in view of Gill et al., (US2018/0244748, filed 7/28/2016, published 8/30/2018, see IDS filed 7/20/2023), in further view of Kariko et al. (Mol Ther, 2008, 16:1833-1840)
As discussed previously, Gill makes a method of modifying macrophages comprising transfection with modified mRNA encoding a CAR.
However, although Gill suggest using nucleotide analogues [0265], they are silent to the mRNA being modified with a pseudouridine.
In regard to claim 4, Kariko teaches methods of expressing mRNA in cells, wherein the mRNA is modified to include psuedouridine (Abstract).
Accordingly, it would have been prima facie obvious to one of ordinary skill in the art at the time of filing to claim the method of modifying macrophages comprising transfection with modified CAR mRNA as taught by Gill and substitute modified nucleosides such as a pseudouridine as taught by Kariko with a reasonable expectation of success. The ordinary skilled artisan would have been motivated to do so as taught by Kariko because these modified mRNAs have higher translational capacity than unmodified mRNAs (Abstract).
Since the instant application claims are obvious over cited patent claims in view of Gill et al., said claims are not patentably distinct.
Claims 17 and 20 are rejected on the grounds of nonstatutory double patenting over 17-19 of U.S. Patent No. 11,332,511 (Gill, Klichinsky, and June, Patented 5/17/2022), in view of Gill et al., (US2018/0244748, filed 7/28/2016, published 8/30/2018, see IDS filed 7/20/2023), in further view of Sahin et al. (WO2014/072061, filed 11/07/2013, published 5/15/2014)
As discussed previously, Gill makes obvious a method of modifying macrophages comprising transfection with modified mRNA encoding CAR
However, Gill is silent with respect to further including step of treating the cells with an RNAaseL inhibitor.
Sahir teaches methods of overexpressing IVT modified RNA in cells including macrophages (Abstract, p. 67, 1st para., see Claims 1-5 of Sahir).
In regard to claim 17, Sahin teaches the steps of inhibiting intracellular IFN signaling by inhibiting RNAseL (p. 3, 4th para., p. 49, 1st para., see Claims 12-15 and 30 of Sahir).
In regard to claim 20, Sahin teaches the cells are treated with the inhibitor prior to delivering the mRNA encoding the protein to be expressed (p. 50, 4th para.).
Accordingly, it would have been prima facie obvious to one of ordinary skill in the art at the time of filing to claim the method of modifying macrophages comprising transfection with modified mRNA encoding CAR as taught by Gill and to combine step of inhibiting RNaseL as taught by Sahin with a reasonable expectation of success. The ordinary skilled artisan would have been motivated to do so as taught by Sahin because if the intracellular IFN signaling mediated by RNaseL is not inhibited, then RNA degradation occurs and/or the inhibition of RNA translation (p. 3, 4th para., p. 49, 1st para.). In regard to choosing to treat the cells with the RNaseL inhibitor before RNA delivery, it would have been obvious to choose the pretreatment option so as to avoid mRNA degradation upon delivery.
Since the instant application claims are obvious over cited patent claims in view of Gill et al., said claims are not patentably distinct.
Claim 18 is rejected are rejected on the grounds of nonstatutory double patenting over 17-19 of U.S. Patent No. 11,332,511 (Gill, Klichinsky, and June, Patented 5/17/2022), in view of Gill et al., (US2018/0244748, filed 7/28/2016, published 8/30/2018, see IDS filed 7/20/2023) and Sahin et al. (WO2014/072061, filed 11/07/2013, published 5/15/2014, in further view of Jha et al. (JBC, 2011, 286:26319-26326)
As discussed previously, Gill in view of Sahin make obvious a method of modifying macrophages comprising transfection with modified mRNA encoding CAR and inhibiting RNaseL.
However, although Sahir teaches the inhibitor of IFN intracellular signaling is an imidazole-oxindole compound, Gill et al. are silent with respect the RNAaseL inhibitor as the oxindole compound of sinitunib.
Jha teaches that sunitinib is a potent inhibitor of RNaseL in cultured mammalian cells (Abstract, Introduction).
Accordingly, it would have been prima facie obvious to one of ordinary skill in the art at the time of filing to claim the method of modifying macrophages with a CAR mRNA and further combine an RNaseL inhibitor to improve mRNA expression as suggested by Gill and Sahin and choose sunitinib as the RNaseL inhibitor as taught by Jha with a reasonable expectation of success. The ordinary skilled artisan would have been motivated to do so as taught by Jha because of the in vitro potency of sunitinib in cultured mammalian cells.
Since the instant application claims are obvious over cited patent claims in view of Gill et al., said claims are not patentably distinct.
Claims 21-24 are rejected on the grounds of nonstatutory double patenting over 17-19 of U.S. Patent No. 11,332,511 (Gill, Klichinsky, and June, Patented 5/17/2022)), in view of Gill et al., (US2018/0244748, filed 7/28/2016, published 8/30/2018, see IDS filed 7/20/2023), in view of Xie et al. (IJMM, 2016, 38:148-160)
As discussed previously, Gill make obvious a method of modifying macrophages comprising transfection with modified mRNA encoding CAR.
However, although Gill suggests the method steps of stimulating the CAR monocytes (i.e., CARMA) by treatments with synergistic compounds such as cytokines after delivering of the CAR nucleic acid ([0127, 0384], Fig. 28), and Gill discloses the ability of macrophages to become polarized, and teaches the anti-tumor advantages of up-regulation of M1 markers in inflammatory/activated macrophages [0012, 0283, 0374], see also Claims 1 and 9 of Gill), Gill does not reduce to practice the step of culturing the RNA transfected macrophages with IFN-beta.
Xie teaches methods of treating macrophages with cytokines in order to induce M1 polarization and teaches that media comprising IFN-beta is required for sustained M1 polarization (Abstract a chimeric receptors (Abstract).
Accordingly, it would have been prima facie obvious to one of ordinary skill in the art at the time of filing to claim the method of modifying macrophages comprising transfection with modified mRNA encoding CAR and to treat the CARMA with cytokines to induce a M1 phenotype as suggested by Gill and to combine IFN-beta in the cytokine treatment as suggested by Xie with a reasonable expectation of success. The ordinary skilled artisan would have been motivated to do so as taught by Xie because IFN-beta plays a crucial role in the regulation of M1 polarization of macrophages (Abstract, Discussion, p. 158).
Since the instant application claims are obvious over cited patent claims in view of Gill et al., said claims are not patentably distinct.
Claims 1, 2, 5, 6, 21-22, and 25-27 are rejected on the grounds of nonstatutory double patenting over claims 15-17 of U.S. Patent No. 11,325,963 (Gill, Klichinsky, and June, Patented 5/10/2022) in view of Gill et al., (US2018/0244748, filed 7/28/2016, published 8/30/2018, see IDS filed 7/20/2023).
The subject matter claimed in the instant application is fully disclosed in the referenced patent as follows: the method for modifying a macrophage or monocyte comprising delivering a mRNA that encodes a CAR of cited patent makes obvious the method of instant application. It is clear that all the elements of the cited patent claims are to be found in instant claims. The difference between the cited patent claims and the instant claims lies in the fact that the instant claims recite that the mRNA acid is a purified modified mRNA.
In regard to claim 1, Gill teaches an in vitro method of modifying human macrophages comprising the following steps:
Modifying a mRNA encoding a CAR produced by a mMessage mMachine T7 ultras In Vitro Transcription kit,
Purifying the mRNA using a Qiagen RNEasy RNA purification kit, and
Delivering the mRNA into the macrophages by electroporation ([0325], Figs 12 & 13, see also Claims 18-20 of Gill).
In regard to claim 2, Gill teaches the mRNA is modified have both a cap on the 5’end and a 3’ poly(A) tail which determine ribosome binding, initiation of translation and stability of the mRNA [0267, 0272].
Accordingly it would have been obvious to claim the cited method and to further claim a purified modified mRNA as taught by Gill with a reasonable expectation of success. One of ordinary skill in the art would have claimed so because of the high efficiency of CAR transfection with purified modified RNA.
In regard to claim 5, as stated supra, Gill teaches the purification step using the RNA using a Qiagen RNEasy RNA purification kit, which comprises a silica membrane purification column.
In regard to claim 6, as stated supra, Gill teaches transfection by electroporation in order to achieve high efficiency transfection.
In regard to claims 21, 22, and 25, Gill teaches the CAR macrophages are cultured for 30 min with beta-glucan and then tested in an in vitro killing assay [0326], which would have been obvious to claim so as to test the efficacy of the CARMA.
In regard to claim 25, as stated supra, Gill teaches the macrophages express the CAR.
In regard to claims 26 and 27, as stated supra, Gill teaches the mRNA is modified with a 5’cap and a 3’ poly(A) tail which contributed to stability of the mRNA.
Since the instant application claims are obvious over cited patent claims in view of Gill, said claims are not patentably distinct.
Claim 3 is rejected on the grounds of nonstatutory double patenting over claims 15-17 of U.S. Patent No. 11,325,963 (Gill, Klichinsky, and June, Patented 5/10/2022), in view of Gill et al., (US2018/0244748, filed 7/28/2016, published 8/30/2018, see IDS filed 7/20/2023), in view further of Grudzien-Nogalska (Meth Enzym, 2007, 431:203-227)
As discussed previously, Gill makes obvious an in vitro method of modifying macrophages comprising transfection with modified mRNA encoding a CAR.
However, although Gill makes obvious the mRNA is modified such as with a 5’ cap, they are silent to an ARAC 5’ cap.
In regard to claim 3, Grudzien-Nogalska reviews ARACs and their applications in mRNA translation and stability (p. 204-224).
Accordingly, it would have been prima facie obvious to one of ordinary skill in the art at the time of filing to claim the method of modifying macrophages comprising transfection with modified CAR mRNA comprising a 5’cap as taught by Gill and to choose the ARAC as taught by Gurdzien-Nogalska with a reasonable expectation of success. The ordinary skilled artisan would have been motivated to do so as taught by Gurdzien-Nogalska because ARACs show two-fold higher translational efficiency compared to conventional caps (p. 205, Introduction).
Since the instant application claims are obvious over cited patent claims in view of Gill et al., said claims are not patentably distinct.
Claim 4 is rejected on the grounds of nonstatutory double patenting over claims 15-17 of U.S. Patent No. 11,325,963 (Gill, Klichinsky, and June, Patented 5/10/2022), in view of Gill et al., (US2018/0244748, filed 7/28/2016, published 8/30/2018, see IDS filed 7/20/2023), in further view of Kariko et al. (Mol Ther, 2008, 16:1833-1840)
As discussed previously, Gill makes a method of modifying macrophages comprising transfection with modified mRNA encoding a CAR.
However, although Gill suggest using nucleotide analogues [0265], they are silent to the mRNA being modified with a pseudouridine.
In regard to claim 4, Kariko teaches methods of expressing mRNA in cells, wherein the mRNA is modified to include psuedouridine (Abstract).
Accordingly, it would have been prima facie obvious to one of ordinary skill in the art at the time of filing to claim the method of modifying macrophages comprising transfection with modified CAR mRNA as taught by Gill and substitute modified nucleosides such as a pseudouridine as taught by Kariko with a reasonable expectation of success. The ordinary skilled artisan would have been motivated to do so as taught by Kariko because these modified mRNAs have higher translational capacity than unmodified mRNAs (Abstract).
Since the instant application claims are obvious over cited patent claims in view of Gill et al., said claims are not patentably distinct.
Claims 17 and 20 are rejected on the grounds of nonstatutory double patenting over claims 15-17 of U.S. Patent No. 11,325,963 (Gill, Klichinsky, and June, Patented 5/10/2022), in view of Gill et al., (US2018/0244748, filed 7/28/2016, published 8/30/2018, see IDS filed 7/20/2023), in further view of Sahin et al. (WO2014/072061, filed 11/07/2013, published 5/15/2014)
As discussed previously, Gill makes obvious a method of modifying macrophages comprising transfection with modified mRNA encoding CAR
However, Gill is silent with respect to further including step of treating the cells with an RNAaseL inhibitor.
Sahir teaches methods of overexpressing IVT modified RNA in cells including macrophages (Abstract, p. 67, 1st para., see Claims 1-5 of Sahir).
In regard to claim 17, Sahin teaches the steps of inhibiting intracellular IFN signaling by inhibiting RNAseL (p. 3, 4th para., p. 49, 1st para., see Claims 12-15 and 30 of Sahir).
In regard to claim 20, Sahin teaches the cells are treated with the inhibitor prior to delivering the mRNA encoding the protein to be expressed (p. 50, 4th para.).
Accordingly, it would have been prima facie obvious to one of ordinary skill in the art at the time of filing to claim the method of modifying macrophages comprising transfection with modified mRNA encoding CAR as taught by Gill and to combine step of inhibiting RNaseL as taught by Sahin with a reasonable expectation of success. The ordinary skilled artisan would have been motivated to do so as taught by Sahin because if the intracellular IFN signaling mediated by RNaseL is not inhibited, then RNA degradation occurs and/or the inhibition of RNA translation (p. 3, 4th para., p. 49, 1st para.). In regard to choosing to treat the cells with the RNaseL inhibitor before RNA delivery, it would have been obvious to choose the pretreatment option so as to avoid mRNA degradation upon delivery.
Since the instant application claims are obvious over cited patent claims in view of Gill et al., said claims are not patentably distinct.
Claim 18 is rejected are rejected on the grounds of nonstatutory double patenting over claims 15-17 of U.S. Patent No. 11,325,963 (Gill, Klichinsky, and June, Patented 5/10/2022), in view of Gill et al., (US2018/0244748, filed 7/28/2016, published 8/30/2018, see IDS filed 7/20/2023) and Sahin et al. (WO2014/072061, filed 11/07/2013, published 5/15/2014, in further view of Jha et al. (JBC, 2011, 286:26319-26326)
As discussed previously, Gill in view of Sahin make obvious a method of modifying macrophages comprising transfection with modified mRNA encoding CAR and inhibiting RNaseL.
However, although Sahir teaches the inhibitor of IFN intracellular signaling is an imidazole-oxindole compound, Gill et al. are silent with respect the RNAaseL inhibitor as the oxindole compound of sinitunib.
Jha teaches that sunitinib is a potent inhibitor of RNaseL in cultured mammalian cells (Abstract, Introduction).
Accordingly, it would have been prima facie obvious to one of ordinary skill in the art at the time of filing to claim the method of modifying macrophages with a CAR mRNA and further combine an RNaseL inhibitor to improve mRNA expression as suggested by Gill and Sahin and choose sunitinib as the RNaseL inhibitor as taught by Jha with a reasonable expectation of success. The ordinary skilled artisan would have been motivated to do so as taught by Jha because of the in vitro potency of sunitinib in cultured mammalian cells.
Since the instant application claims are obvious over cited patent claims in view of Gill et al., said claims are not patentably distinct.
Claims 21-24 are rejected on the grounds of nonstatutory double patenting over claims 15-17 of U.S. Patent No. 11,325,963 (Gill, Klichinsky, and June, Patented 5/10/2022), in view of Gill et al., (US2018/0244748, filed 7/28/2016, published 8/30/2018, see IDS filed 7/20/2023), in view of Xie et al. (IJMM, 2016, 38:148-160)
As discussed previously, Gill make obvious a method of modifying macrophages comprising transfection with modified mRNA encoding CAR.
However, although Gill suggests the method steps of stimulating the CAR monocytes (i.e., CARMA) by treatments with synergistic compounds such as cytokines after delivering of the CAR nucleic acid ([0127, 0384], Fig. 28), and Gill discloses the ability of macrophages to become polarized, and teaches the anti-tumor advantages of up-regulation of M1 markers in inflammatory/activated macrophages [0012, 0283, 0374], see also Claims 1 and 9 of Gill), Gill does not reduce to practice the step of culturing the RNA transfected macrophages with IFN-beta.
Xie teaches methods of treating macrophages with cytokines in order to induce M1 polarization and teaches that media comprising IFN-beta is required for sustained M1 polarization (Abstract a chimeric receptors (Abstract).
Accordingly, it would have been prima facie obvious to one of ordinary skill in the art at the time of filing to claim the method of modifying macrophages comprising transfection with modified mRNA encoding CAR and to treat the CARMA with cytokines to induce a M1 phenotype as suggested by Gill and to combine IFN-beta in the cytokine treatment as suggested by Xie with a reasonable expectation of success. The ordinary skilled artisan would have been motivated to do so as taught by Xie because IFN-beta plays a crucial role in the regulation of M1 polarization of macrophages (Abstract, Discussion, p. 158).
Since the instant application claims are obvious over cited patent claims in view of Gill et al., said claims are not patentably distinct.
Claims 1, 2, 5, 6, 21-22, and 25-27 are rejected on the grounds of nonstatutory double patenting over claims 15-17 of U.S. Patent No. 11,319,358 (Gill, Klichinsky, and June, Patented 5/03/2022) in view of Gill et al., (US2018/0244748, filed 7/28/2016, published 8/30/2018, see IDS filed 7/20/2023).
The subject matter claimed in the instant application is fully disclosed in the referenced patent as follows: the method for modifying a macrophage or monocyte comprising delivering a mRNA that encodes a CAR of cited patent makes obvious the method of instant application. It is clear that all the elements of the cited patent claims are to be found in instant claims. The difference between the cited patent claims and the instant claims lies in the fact that the instant claims recite that the mRNA acid is a purified modified mRNA.
In regard to claim 1, Gill teaches an in vitro method of modifying human macrophages comprising the following steps:
Modifying a mRNA encoding a CAR produced by a mMessage mMachine T7 ultras In Vitro Transcription kit,
Purifying the mRNA using a Qiagen RNEasy RNA purification kit, and
Delivering the mRNA into the macrophages by electroporation ([0325], Figs 12 & 13, see also Claims 18-20 of Gill).
In regard to claim 2, Gill teaches the mRNA is modified have both a cap on the 5’end and a 3’ poly(A) tail which determine ribosome binding, initiation of translation and stability of the mRNA [0267, 0272].
Accordingly it would have been obvious to claim the cited method and to further claim a purified modified mRNA as taught by Gill with a reasonable expectation of success. One of ordinary skill in the art would have claimed so because of the high efficiency of CAR transfection with purified modified RNA.
In regard to claim 5, as stated supra, Gill teaches the purification step using the RNA using a Qiagen RNEasy RNA purification kit, which comprises a silica membrane purification column.
In regard to claim 6, as stated supra, Gill teaches transfection by electroporation in order to achieve high efficiency transfection.
In regard to claims 21, 22, and 25, Gill teaches the CAR macrophages are cultured for 30 min with beta-glucan and then tested in an in vitro killing assay [0326], which would have been obvious to claim so as to test the efficacy of the CARMA.
In regard to claim 25, as stated supra, Gill teaches the macrophages express the CAR.
In regard to claims 26 and 27, as stated supra, Gill teaches the mRNA is modified with a 5’cap and a 3’ poly(A) tail which contributed to stability of the mRNA.
Since the instant application claims are obvious over cited patent claims in view of Gill, said claims are not patentably distinct.
Claim 3 is rejected on the grounds of nonstatutory double patenting over claims 15-17 of U.S. Patent No. 11,319,358 (Gill, Klichinsky, and June, Patented 5/03/2022), in view of Gill et al., (US2018/0244748, filed 7/28/2016, published 8/30/2018, see IDS filed 7/20/2023), in view further of Grudzien-Nogalska (Meth Enzym, 2007, 431:203-227)
As discussed previously, Gill makes obvious an in vitro method of modifying macrophages comprising transfection with modified mRNA encoding a CAR.
However, although Gill makes obvious the mRNA is modified such as with a 5’ cap, they are silent to an ARAC 5’ cap.
In regard to claim 3, Grudzien-Nogalska reviews ARACs and their applications in mRNA translation and stability (p. 204-224).
Accordingly, it would have been prima facie obvious to one of ordinary skill in the art at the time of filing to claim the method of modifying macrophages comprising transfection with modified CAR mRNA comprising a 5’cap as taught by Gill and to choose the ARAC as taught by Gurdzien-Nogalska with a reasonable expectation of success. The ordinary skilled artisan would have been motivated to do so as taught by Gurdzien-Nogalska because ARACs show two-fold higher translational efficiency compared to conventional caps (p. 205, Introduction).
Since the instant application claims are obvious over cited patent claims in view of Gill et al., said claims are not patentably distinct.
Claim 4 is rejected on the grounds of nonstatutory double patenting over claims 15-17 of U.S. Patent No. 11,319,358 (Gill, Klichinsky, and June, Patented 5/03/2022), in view of Gill et al., (US2018/0244748, filed 7/28/2016, published 8/30/2018, see IDS filed 7/20/2023), in further view of Kariko et al. (Mol Ther, 2008, 16:1833-1840)
As discussed previously, Gill makes a method of modifying macrophages comprising transfection with modified mRNA encoding a CAR.
However, although Gill suggest using nucleotide analogues [0265], they are silent to the mRNA being modified with a pseudouridine.
In regard to claim 4, Kariko teaches methods of expressing mRNA in cells, wherein the mRNA is modified to include psuedouridine (Abstract).
Accordingly, it would have been prima facie obvious to one of ordinary skill in the art at the time of filing to claim the method of modifying macrophages comprising transfection with modified CAR mRNA as taught by Gill and substitute modified nucleosides such as a pseudouridine as taught by Kariko with a reasonable expectation of success. The ordinary skilled artisan would have been motivated to do so as taught by Kariko because these modified mRNAs have higher translational capacity than unmodified mRNAs (Abstract).
Since the instant application claims are obvious over cited patent claims in view of Gill et al., said claims are not patentably distinct.
Claims 17 and 20 are rejected on the grounds of nonstatutory double patenting over claims 15-17 of U.S. Patent No. 11,319,358 (Gill, Klichinsky, and June, Patented 5/03/2022), in view of Gill et al., (US2018/0244748, filed 7/28/2016, published 8/30/2018, see IDS filed 7/20/2023), in further view of Sahin et al. (WO2014/072061, filed 11/07/2013, published 5/15/2014)
As discussed previously, Gill makes obvious a method of modifying macrophages comprising transfection with modified mRNA encoding CAR
However, Gill is silent with respect to further including step of treating the cells with an RNAaseL inhibitor.
Sahir teaches methods of overexpressing IVT modified RNA in cells including macrophages (Abstract, p. 67, 1st para., see Claims 1-5 of Sahir).
In regard to claim 17, Sahin teaches the steps of inhibiting intracellular IFN signaling by inhibiting RNAseL (p. 3, 4th para., p. 49, 1st para., see Claims 12-15 and 30 of Sahir).
In regard to claim 20, Sahin teaches the cells are treated with the inhibitor prior to delivering the mRNA encoding the protein to be expressed (p. 50, 4th para.).
Accordingly, it would have been prima facie obvious to one of ordinary skill in the art at the time of filing to claim the method of modifying macrophages comprising transfection with modified mRNA encoding CAR as taught by Gill and to combine step of inhibiting RNaseL as taught by Sahin with a reasonable expectation of success. The ordinary skilled artisan would have been motivated to do so as taught by Sahin because if the intracellular IFN signaling mediated by RNaseL is not inhibited, then RNA degradation occurs and/or the inhibition of RNA translation (p. 3, 4th para., p. 49, 1st para.). In regard to choosing to treat the cells with the RNaseL inhibitor before RNA delivery, it would have been obvious to choose the pretreatment option so as to avoid mRNA degradation upon delivery.
Since the instant application claims are obvious over cited patent claims in view of Gill et al., said claims are not patentably distinct.
Claim 18 is rejected are rejected on the grounds of nonstatutory double patenting over claims 15-17 of U.S. Patent No. 11,319,358 (Gill, Klichinsky, and June, Patented 5/03/2022), in view of Gill et al., (US2018/0244748, filed 7/28/2016, published 8/30/2018, see IDS filed 7/20/2023) and Sahin et al. (WO2014/072061, filed 11/07/2013, published 5/15/2014, in further view of Jha et al. (JBC, 2011, 286:26319-26326)
As discussed previously, Gill in view of Sahin make obvious a method of modifying macrophages comprising transfection with modified mRNA encoding CAR and inhibiting RNaseL.
However, although Sahir teaches the inhibitor of IFN intracellular signaling is an imidazole-oxindole compound, Gill et al. are silent with respect the RNAaseL inhibitor as the oxindole compound of sinitunib.
Jha teaches that sunitinib is a potent inhibitor of RNaseL in cultured mammalian cells (Abstract, Introduction).
Accordingly, it would have been prima facie obvious to one of ordinary skill in the art at the time of filing to claim the method of modifying macrophages with a CAR mRNA and further combine an RNaseL inhibitor to improve mRNA expression as suggested by Gill and Sahin and choose sunitinib as the RNaseL inhibitor as taught by Jha with a reasonable expectation of success. The ordinary skilled artisan would have been motivated to do so as taught by Jha because of the in vitro potency of sunitinib in cultured mammalian cells.
Since the instant application claims are obvious over cited patent claims in view of Gill et al., said claims are not patentably distinct.
Claims 21-24 are rejected on the grounds of nonstatutory double patenting over claims 15-17 of U.S. Patent No. 11,319,358 (Gill, Klichinsky, and June, Patented 5/03/2022), in view of Gill et al., (US2018/0244748, filed 7/28/2016, published 8/30/2018, see IDS filed 7/20/2023), in view of Xie et al. (IJMM, 2016, 38:148-160)
As discussed previously, Gill make obvious a method of modifying macrophages comprising transfection with modified mRNA encoding CAR.
However, although Gill suggests the method steps of stimulating the CAR monocytes (i.e., CARMA) by treatments with synergistic compounds such as cytokines after delivering of the CAR nucleic acid ([0127, 0384], Fig. 28), and Gill discloses the ability of macrophages to become polarized, and teaches the anti-tumor advantages of up-regulation of M1 markers in inflammatory/activated macrophages [0012, 0283, 0374], see also Claims 1 and 9 of Gill), Gill does not reduce to practice the step of culturing the RNA transfected macrophages with IFN-beta.
Xie teaches methods of treating macrophages with cytokines in order to induce M1 polarization and teaches that media comprising IFN-beta is required for sustained M1 polarization (Abstract a chimeric receptors (Abstract).
Accordingly, it would have been prima facie obvious to one of ordinary skill in the art at the time of filing to claim the method of modifying macrophages comprising transfection with modified mRNA encoding CAR and to treat the CARMA with cytokines to induce a M1 phenotype as suggested by Gill and to combine IFN-beta in the cytokine treatment as suggested by Xie with a reasonable expectation of success. The ordinary skilled artisan would have been motivated to do so as taught by Xie because IFN-beta plays a crucial role in the regulation of M1 polarization of macrophages (Abstract, Discussion, p. 158).
Since the instant application claims are obvious over cited patent claims in view of Gill et al., said claims are not patentably distinct.
Claims 1, 2, 5, 6, 21-22, and 25-27 are rejected on the grounds of nonstatutory double patenting over claims 17-19 of U.S. Patent No. 11,306,134 (Gill, Klichinsky, and June, Patented 4/19/2022) in view of Gill et al., (US2018/0244748, filed 7/28/2016, published 8/30/2018, see IDS filed 7/20/2023).
The subject matter claimed in the instant application is fully disclosed in the referenced patent as follows: the method for modifying a macrophage or monocyte comprising delivering a mRNA that encodes a CAR of cited patent makes obvious the method of instant application. It is clear that all the elements of the cited patent claims are to be found in instant claims. The difference between the cited patent claims and the instant claims lies in the fact that the instant claims recite that the mRNA acid is a purified modified mRNA.
In regard to claim 1, Gill teaches an in vitro method of modifying human macrophages comprising the following steps:
Modifying a mRNA encoding a CAR produced by a mMessage mMachine T7 ultras In Vitro Transcription kit,
Purifying the mRNA using a Qiagen RNEasy RNA purification kit, and
Delivering the mRNA into the macrophages by electroporation ([0325], Figs 12 & 13, see also Claims 18-20 of Gill).
In regard to claim 2, Gill teaches the mRNA is modified have both a cap on the 5’end and a 3’ poly(A) tail which determine ribosome binding, initiation of translation and stability of the mRNA [0267, 0272].
Accordingly it would have been obvious to claim the cited method and to further claim a purified modified mRNA as taught by Gill with a reasonable expectation of success. One of ordinary skill in the art would have claimed so because of the high efficiency of CAR transfection with purified modified RNA.
In regard to claim 5, as stated supra, Gill teaches the purification step using the RNA using a Qiagen RNEasy RNA purification kit, which comprises a silica membrane purification column.
In regard to claim 6, as stated supra, Gill teaches transfection by electroporation in order to achieve high efficiency transfection.
In regard to claims 21, 22, and 25, Gill teaches the CAR macrophages are cultured for 30 min with beta-glucan and then tested in an in vitro killing assay [0326], which would have been obvious to claim so as to test the efficacy of the CARMA.
In regard to claim 25, as stated supra, Gill teaches the macrophages express the CAR.
In regard to claims 26 and 27, as stated supra, Gill teaches the mRNA is modified with a 5’cap and a 3’ poly(A) tail which contributed to stability of the mRNA.
Since the instant application claims are obvious over cited patent claims in view of Gill, said claims are not patentably distinct.
Claim 3 is rejected on the grounds of nonstatutory double patenting over claims 17-19 of U.S. Patent No. 11,306,134 (Gill, Klichinsky, and June, Patented 4/19/2022), in view of Gill et al., (US2018/0244748, filed 7/28/2016, published 8/30/2018, see IDS filed 7/20/2023), in view further of Grudzien-Nogalska (Meth Enzym, 2007, 431:203-227)
As discussed previously, Gill makes obvious an in vitro method of modifying macrophages comprising transfection with modified mRNA encoding a CAR.
However, although Gill makes obvious the mRNA is modified such as with a 5’ cap, they are silent to an ARAC 5’ cap.
In regard to claim 3, Grudzien-Nogalska reviews ARACs and their applications in mRNA translation and stability (p. 204-224).
Accordingly, it would have been prima facie obvious to one of ordinary skill in the art at the time of filing to claim the method of modifying macrophages comprising transfection with modified CAR mRNA comprising a 5’cap as taught by Gill and to choose the ARAC as taught by Gurdzien-Nogalska with a reasonable expectation of success. The ordinary skilled artisan would have been motivated to do so as taught by Gurdzien-Nogalska because ARACs show two-fold higher translational efficiency compared to conventional caps (p. 205, Introduction).
Since the instant application claims are obvious over cited patent claims in view of Gill et al., said claims are not patentably distinct.
Claim 4 is rejected on the grounds of nonstatutory double patenting over claims 17-19 of U.S. Patent No. 11,306,134 (Gill, Klichinsky, and June, Patented 4/19/2022), in view of Gill et al., (US2018/0244748, filed 7/28/2016, published 8/30/2018, see IDS filed 7/20/2023), in further view of Kariko et al. (Mol Ther, 2008, 16:1833-1840)
As discussed previously, Gill makes a method of modifying macrophages comprising transfection with modified mRNA encoding a CAR.
However, although Gill suggest using nucleotide analogues [0265], they are silent to the mRNA being modified with a pseudouridine.
In regard to claim 4, Kariko teaches methods of expressing mRNA in cells, wherein the mRNA is modified to include psuedouridine (Abstract).
Accordingly, it would have been prima facie obvious to one of ordinary skill in the art at the time of filing to claim the method of modifying macrophages comprising transfection with modified CAR mRNA as taught by Gill and substitute modified nucleosides such as a pseudouridine as taught by Kariko with a reasonable expectation of success. The ordinary skilled artisan would have been motivated to do so as taught by Kariko because these modified mRNAs have higher translational capacity than unmodified mRNAs (Abstract).
Since the instant application claims are obvious over cited patent claims in view of Gill et al., said claims are not patentably distinct.
Claims 17 and 20 are rejected on the grounds of nonstatutory double patenting over claims 17-19 of U.S. Patent No. 11,306,134 (Gill, Klichinsky, and June, Patented 4/19/2022), in view of Gill et al., (US2018/0244748, filed 7/28/2016, published 8/30/2018, see IDS filed 7/20/2023), in further view of Sahin et al. (WO2014/072061, filed 11/07/2013, published 5/15/2014)
As discussed previously, Gill makes obvious a method of modifying macrophages comprising transfection with modified mRNA encoding CAR
However, Gill is silent with respect to further including step of treating the cells with an RNAaseL inhibitor.
Sahir teaches methods of overexpressing IVT modified RNA in cells including macrophages (Abstract, p. 67, 1st para., see Claims 1-5 of Sahir).
In regard to claim 17, Sahin teaches the steps of inhibiting intracellular IFN signaling by inhibiting RNAseL (p. 3, 4th para., p. 49, 1st para., see Claims 12-15 and 30 of Sahir).
In regard to claim 20, Sahin teaches the cells are treated with the inhibitor prior to delivering the mRNA encoding the protein to be expressed (p. 50, 4th para.).
Accordingly, it would have been prima facie obvious to one of ordinary skill in the art at the time of filing to claim the method of modifying macrophages comprising transfection with modified mRNA encoding CAR as taught by Gill and to combine step of inhibiting RNaseL as taught by Sahin with a reasonable expectation of success. The ordinary skilled artisan would have been motivated to do so as taught by Sahin because if the intracellular IFN signaling mediated by RNaseL is not inhibited, then RNA degradation occurs and/or the inhibition of RNA translation (p. 3, 4th para., p. 49, 1st para.). In regard to choosing to treat the cells with the RNaseL inhibitor before RNA delivery, it would have been obvious to choose the pretreatment option so as to avoid mRNA degradation upon delivery.
Since the instant application claims are obvious over cited patent claims in view of Gill et al., said claims are not patentably distinct.
Claim 18 is rejected are rejected on the grounds of nonstatutory double patenting over claims 17-19 of U.S. Patent No. 11,306,134 (Gill, Klichinsky, and June, Patented 4/19/2022), in view of Gill et al., (US2018/0244748, filed 7/28/2016, published 8/30/2018, see IDS filed 7/20/2023) and Sahin et al. (WO2014/072061, filed 11/07/2013, published 5/15/2014, in further view of Jha et al. (JBC, 2011, 286:26319-26326)
As discussed previously, Gill in view of Sahin make obvious a method of modifying macrophages comprising transfection with modified mRNA encoding CAR and inhibiting RNaseL.
However, although Sahir teaches the inhibitor of IFN intracellular signaling is an imidazole-oxindole compound, Gill et al. are silent with respect the RNAaseL inhibitor as the oxindole compound of sinitunib.
Jha teaches that sunitinib is a potent inhibitor of RNaseL in cultured mammalian cells (Abstract, Introduction).
Accordingly, it would have been prima facie obvious to one of ordinary skill in the art at the time of filing to claim the method of modifying macrophages with a CAR mRNA and further combine an RNaseL inhibitor to improve mRNA expression as suggested by Gill and Sahin and choose sunitinib as the RNaseL inhibitor as taught by Jha with a reasonable expectation of success. The ordinary skilled artisan would have been motivated to do so as taught by Jha because of the in vitro potency of sunitinib in cultured mammalian cells.
Since the instant application claims are obvious over cited patent claims in view of Gill et al., said claims are not patentably distinct.
Claims 21-24 are rejected on the grounds of nonstatutory double patenting over claims 17-19 of U.S. Patent No. 11,306,134 (Gill, Klichinsky, and June, Patented 4/19/2022), in view of Gill et al., (US2018/0244748, filed 7/28/2016, published 8/30/2018, see IDS filed 7/20/2023), in view of Xie et al. (IJMM, 2016, 38:148-160)
As discussed previously, Gill make obvious a method of modifying macrophages comprising transfection with modified mRNA encoding CAR.
However, although Gill suggests the method steps of stimulating the CAR monocytes (i.e., CARMA) by treatments with synergistic compounds such as cytokines after delivering of the CAR nucleic acid ([0127, 0384], Fig. 28), and Gill discloses the ability of macrophages to become polarized, and teaches the anti-tumor advantages of up-regulation of M1 markers in inflammatory/activated macrophages [0012, 0283, 0374], see also Claims 1 and 9 of Gill), Gill does not reduce to practice the step of culturing the RNA transfected macrophages with IFN-beta.
Xie teaches methods of treating macrophages with cytokines in order to induce M1 polarization and teaches that media comprising IFN-beta is required for sustained M1 polarization (Abstract a chimeric receptors (Abstract).
Accordingly, it would have been prima facie obvious to one of ordinary skill in the art at the time of filing to claim the method of modifying macrophages comprising transfection with modified mRNA encoding CAR and to treat the CARMA with cytokines to induce a M1 phenotype as suggested by Gill and to combine IFN-beta in the cytokine treatment as suggested by Xie with a reasonable expectation of success. The ordinary skilled artisan would have been motivated to do so as taught by Xie because IFN-beta plays a crucial role in the regulation of M1 polarization of macrophages (Abstract, Discussion, p. 158).
Since the instant application claims are obvious over cited patent claims in view of Gill et al., said claims are not patentably distinct.
Claims 1, 2, 5, 6, 21-22, and 25-27 are rejected on the grounds of nonstatutory double patenting over claims 16-18 and 27 of U.S. Patent No. 11,034,749 (Gill, Klichinsky, and June, Patented 6/15/2021) in view of Gill et al., (US2018/0244748, filed 7/28/2016, published 8/30/2018, see IDS filed 7/20/2023).
The subject matter claimed in the instant application is fully disclosed in the referenced patent as follows: the method for modifying a macrophage or monocyte comprising delivering a mRNA that encodes a CAR of cited patent makes obvious the method of instant application. It is clear that all the elements of the cited patent claims are to be found in instant claims. The difference between the cited patent claims and the instant claims lies in the fact that the instant claims recite that the mRNA acid is a purified modified mRNA.
In regard to claim 1, Gill teaches an in vitro method of modifying human macrophages comprising the following steps:
Modifying a mRNA encoding a CAR produced by a mMessage mMachine T7 ultras In Vitro Transcription kit,
Purifying the mRNA using a Qiagen RNEasy RNA purification kit, and
Delivering the mRNA into the macrophages by electroporation ([0325], Figs 12 & 13, see also Claims 18-20 of Gill).
In regard to claim 2, Gill teaches the mRNA is modified have both a cap on the 5’end and a 3’ poly(A) tail which determine ribosome binding, initiation of translation and stability of the mRNA [0267, 0272].
Accordingly it would have been obvious to claim the cited method and to further claim a purified modified mRNA as taught by Gill with a reasonable expectation of success. One of ordinary skill in the art would have claimed so because of the high efficiency of CAR transfection with purified modified RNA.
In regard to claim 5, as stated supra, Gill teaches the purification step using the RNA using a Qiagen RNEasy RNA purification kit, which comprises a silica membrane purification column.
In regard to claim 6, as stated supra, Gill teaches transfection by electroporation in order to achieve high efficiency transfection.
In regard to claims 21, 22, and 25, Gill teaches the CAR macrophages are cultured for 30 min with beta-glucan and then tested in an in vitro killing assay [0326], which would have been obvious to claim so as to test the efficacy of the CARMA.
In regard to claim 25, as stated supra, Gill teaches the macrophages express the CAR.
In regard to claims 26 and 27, as stated supra, Gill teaches the mRNA is modified with a 5’cap and a 3’ poly(A) tail which contributed to stability of the mRNA.
Since the instant application claims are obvious over cited patent claims in view of Gill, said claims are not patentably distinct.
Claim 3 is rejected on the grounds of nonstatutory double patenting over claims 16-18 and 27 of U.S. Patent No. 11,034,749 (Gill, Klichinsky, and June, Patented 6/15/2021), in view of Gill et al., (US2018/0244748, filed 7/28/2016, published 8/30/2018, see IDS filed 7/20/2023), in view further of Grudzien-Nogalska (Meth Enzym, 2007, 431:203-227)
As discussed previously, Gill makes obvious an in vitro method of modifying macrophages comprising transfection with modified mRNA encoding a CAR.
However, although Gill makes obvious the mRNA is modified such as with a 5’ cap, they are silent to an ARAC 5’ cap.
In regard to claim 3, Grudzien-Nogalska reviews ARACs and their applications in mRNA translation and stability (p. 204-224).
Accordingly, it would have been prima facie obvious to one of ordinary skill in the art at the time of filing to claim the method of modifying macrophages comprising transfection with modified CAR mRNA comprising a 5’cap as taught by Gill and to choose the ARAC as taught by Gurdzien-Nogalska with a reasonable expectation of success. The ordinary skilled artisan would have been motivated to do so as taught by Gurdzien-Nogalska because ARACs show two-fold higher translational efficiency compared to conventional caps (p. 205, Introduction).
Since the instant application claims are obvious over cited patent claims in view of Gill et al., said claims are not patentably distinct.
Claim 4 is rejected on the grounds of nonstatutory double patenting over claims 16-18 and 27 of U.S. Patent No. 11,034,749 (Gill, Klichinsky, and June, Patented 6/15/2021), in view of Gill et al., (US2018/0244748, filed 7/28/2016, published 8/30/2018, see IDS filed 7/20/2023), in further view of Kariko et al. (Mol Ther, 2008, 16:1833-1840)
As discussed previously, Gill makes a method of modifying macrophages comprising transfection with modified mRNA encoding a CAR.
However, although Gill suggest using nucleotide analogues [0265], they are silent to the mRNA being modified with a pseudouridine.
In regard to claim 4, Kariko teaches methods of expressing mRNA in cells, wherein the mRNA is modified to include psuedouridine (Abstract).
Accordingly, it would have been prima facie obvious to one of ordinary skill in the art at the time of filing to claim the method of modifying macrophages comprising transfection with modified CAR mRNA as taught by Gill and substitute modified nucleosides such as a pseudouridine as taught by Kariko with a reasonable expectation of success. The ordinary skilled artisan would have been motivated to do so as taught by Kariko because these modified mRNAs have higher translational capacity than unmodified mRNAs (Abstract).
Since the instant application claims are obvious over cited patent claims in view of Gill et al., said claims are not patentably distinct.
Claims 17 and 20 are rejected on the grounds of nonstatutory double patenting over claims 16-18 and 27 of U.S. Patent No. 11,034,749 (Gill, Klichinsky, and June, Patented 6/15/2021), in view of Gill et al., (US2018/0244748, filed 7/28/2016, published 8/30/2018, see IDS filed 7/20/2023), in further view of Sahin et al. (WO2014/072061, filed 11/07/2013, published 5/15/2014)
As discussed previously, Gill makes obvious a method of modifying macrophages comprising transfection with modified mRNA encoding CAR
However, Gill is silent with respect to further including step of treating the cells with an RNAaseL inhibitor.
Sahir teaches methods of overexpressing IVT modified RNA in cells including macrophages (Abstract, p. 67, 1st para., see Claims 1-5 of Sahir).
In regard to claim 17, Sahin teaches the steps of inhibiting intracellular IFN signaling by inhibiting RNAseL (p. 3, 4th para., p. 49, 1st para., see Claims 12-15 and 30 of Sahir).
In regard to claim 20, Sahin teaches the cells are treated with the inhibitor prior to delivering the mRNA encoding the protein to be expressed (p. 50, 4th para.).
Accordingly, it would have been prima facie obvious to one of ordinary skill in the art at the time of filing to claim the method of modifying macrophages comprising transfection with modified mRNA encoding CAR as taught by Gill and to combine step of inhibiting RNaseL as taught by Sahin with a reasonable expectation of success. The ordinary skilled artisan would have been motivated to do so as taught by Sahin because if the intracellular IFN signaling mediated by RNaseL is not inhibited, then RNA degradation occurs and/or the inhibition of RNA translation (p. 3, 4th para., p. 49, 1st para.). In regard to choosing to treat the cells with the RNaseL inhibitor before RNA delivery, it would have been obvious to choose the pretreatment option so as to avoid mRNA degradation upon delivery.
Since the instant application claims are obvious over cited patent claims in view of Gill et al., said claims are not patentably distinct.
Claim 18 is rejected are rejected on the grounds of nonstatutory double patenting over claims 16-18 and 27 of U.S. Patent No. 11,034,749 (Gill, Klichinsky, and June, Patented 6/15/2021), in view of Gill et al., (US2018/0244748, filed 7/28/2016, published 8/30/2018, see IDS filed 7/20/2023) and Sahin et al. (WO2014/072061, filed 11/07/2013, published 5/15/2014, in further view of Jha et al. (JBC, 2011, 286:26319-26326)
As discussed previously, Gill in view of Sahin make obvious a method of modifying macrophages comprising transfection with modified mRNA encoding CAR and inhibiting RNaseL.
However, although Sahir teaches the inhibitor of IFN intracellular signaling is an imidazole-oxindole compound, Gill et al. are silent with respect the RNAaseL inhibitor as the oxindole compound of sinitunib.
Jha teaches that sunitinib is a potent inhibitor of RNaseL in cultured mammalian cells (Abstract, Introduction).
Accordingly, it would have been prima facie obvious to one of ordinary skill in the art at the time of filing to claim the method of modifying macrophages with a CAR mRNA and further combine an RNaseL inhibitor to improve mRNA expression as suggested by Gill and Sahin and choose sunitinib as the RNaseL inhibitor as taught by Jha with a reasonable expectation of success. The ordinary skilled artisan would have been motivated to do so as taught by Jha because of the in vitro potency of sunitinib in cultured mammalian cells.
Since the instant application claims are obvious over cited patent claims in view of Gill et al., said claims are not patentably distinct.
Claims 21-24 are rejected on the grounds of nonstatutory double patenting over claims 16-18 and 27 of U.S. Patent No. 11,034,749 (Gill, Klichinsky, and June, Patented 6/15/2021), in view of Gill et al., (US2018/0244748, filed 7/28/2016, published 8/30/2018, see IDS filed 7/20/2023), in view of Xie et al. (IJMM, 2016, 38:148-160)
As discussed previously, Gill make obvious a method of modifying macrophages comprising transfection with modified mRNA encoding CAR.
However, although Gill suggests the method steps of stimulating the CAR monocytes (i.e., CARMA) by treatments with synergistic compounds such as cytokines after delivering of the CAR nucleic acid ([0127, 0384], Fig. 28), and Gill discloses the ability of macrophages to become polarized, and teaches the anti-tumor advantages of up-regulation of M1 markers in inflammatory/activated macrophages [0012, 0283, 0374], see also Claims 1 and 9 of Gill), Gill does not reduce to practice the step of culturing the RNA transfected macrophages with IFN-beta.
Xie teaches methods of treating macrophages with cytokines in order to induce M1 polarization and teaches that media comprising IFN-beta is required for sustained M1 polarization (Abstract a chimeric receptors (Abstract).
Accordingly, it would have been prima facie obvious to one of ordinary skill in the art at the time of filing to claim the method of modifying macrophages comprising transfection with modified mRNA encoding CAR and to treat the CARMA with cytokines to induce a M1 phenotype as suggested by Gill and to combine IFN-beta in the cytokine treatment as suggested by Xie with a reasonable expectation of success. The ordinary skilled artisan would have been motivated to do so as taught by Xie because IFN-beta plays a crucial role in the regulation of M1 polarization of macrophages (Abstract, Discussion, p. 158).
Since the instant application claims are obvious over cited patent claims in view of Gill et al., said claims are not patentably distinct.
Provisional Double Patenting
Claims 1, 2, 5, 6, 21-22, and 25-27 are provisionally rejected on the grounds of nonstatutory double patenting as being unpatentable over claims 18-20 of copending Application No. 19/061,415, in view of Gill et al., (US2018/0244748, filed 7/28/2016, published 8/30/2018, see IDS filed 7/20/2023). This is a provisional nonstatutory double patenting rejection because the patentably indistinct claims have not in fact been patented
The subject matter claimed in the instant application is disclosed in the referenced application as follows: the method for modifying an immune cell comprising introducing mRNA encoding a CAR makes obvious the methd of instant application. It is clear that elements of the cited application claims are to be found in instant claims. The difference between the cited application claims and the instant claims lies in the fact that the instant application claims are more specific because they claim a modified and purified mRNA.
In regard to claim 1, Gill teaches an in vitro method of modifying human macrophages comprising the following steps:
Modifying a mRNA encoding a CAR produced by a mMessage mMachine T7 ultras In Vitro Transcription kit,
Purifying the mRNA using a Qiagen RNEasy RNA purification kit, and
Delivering the mRNA into the macrophages by electroporation ([0325], Figs 12 & 13, see also Claims 18-20 of Gill).
In regard to claim 2, Gill teaches the mRNA is modified have both a cap on the 5’end and a 3’ poly(A) tail which determine ribosome binding, initiation of translation and stability of the mRNA [0267, 0272].
Accordingly it would have been obvious to claim the cited method and to further claim a purified modified mRNA as taught by Gill with a reasonable expectation of success. One of ordinary skill in the art would have claimed so because of the high efficiency of CAR transfection with purified modified RNA.
In regard to claim 5, as stated supra, Gill teaches the purification step using the RNA using a Qiagen RNEasy RNA purification kit, which comprises a silica membrane purification column.
In regard to claim 6, as stated supra, Gill teaches transfection by electroporation in order to achieve high efficiency transfection.
In regard to claims 21, 22, and 25, Gill teaches the CAR macrophages are cultured for 30 min with beta-glucan and then tested in an in vitro killing assay [0326], which would have been obvious to claim so as to test the efficacy of the CARMA.
In regard to claim 25, as stated supra, Gill teaches the macrophages express the CAR.
In regard to claims 26 and 27, as stated supra, Gill teaches the mRNA is modified with a 5’cap and a 3’ poly(A) tail which contributed to stability of the mRNA.
Since the instant application claims are obvious over cited application claims, in view of Gill, said claims are not patentably distinct.
Claim 3 is provisionally rejected on the grounds of nonstatutory double patenting as being unpatentable over claims 18-20 of copending Application No. 19/061,415), in view of Gill et al., (US2018/0244748, filed 7/28/2016, published 8/30/2018, see IDS filed 7/20/2023), in view further of Grudzien-Nogalska (Meth Enzym, 2007, 431:203-227)
As discussed previously, Gill makes obvious an in vitro method of modifying macrophages comprising transfection with modified mRNA encoding a CAR.
However, although Gill makes obvious the mRNA is modified such as with a 5’ cap, they are silent to an ARAC 5’ cap.
In regard to claim 3, Grudzien-Nogalska reviews ARACs and their applications in mRNA translation and stability (p. 204-224).
Accordingly, it would have been prima facie obvious to one of ordinary skill in the art at the time of filing to claim the method of modifying macrophages comprising transfection with modified CAR mRNA comprising a 5’cap as taught by Gill and to choose the ARAC as taught by Gurdzien-Nogalska with a reasonable expectation of success. The ordinary skilled artisan would have been motivated to do so as taught by Gurdzien-Nogalska because ARACs show two-fold higher translational efficiency compared to conventional caps (p. 205, Introduction).
Since the instant application claims are obvious over cited patent claims in view of Gill et al., said claims are not patentably distinct.
Claim 4 is provisionally rejected on the grounds of nonstatutory double patenting as being unpatentable over claims 18-20 of copending Application No. 19/061,415), in view of Gill et al., (US2018/0244748, filed 7/28/2016, published 8/30/2018, see IDS filed 7/20/2023), in further view of Kariko et al. (Mol Ther, 2008, 16:1833-1840)
As discussed previously, Gill makes a method of modifying macrophages comprising transfection with modified mRNA encoding a CAR.
However, although Gill suggest using nucleotide analogues [0265], they are silent to the mRNA being modified with a pseudouridine.
In regard to claim 4, Kariko teaches methods of expressing mRNA in cells, wherein the mRNA is modified to include psuedouridine (Abstract).
Accordingly, it would have been prima facie obvious to one of ordinary skill in the art at the time of filing to claim the method of modifying macrophages comprising transfection with modified CAR mRNA as taught by Gill and substitute modified nucleosides such as a pseudouridine as taught by Kariko with a reasonable expectation of success. The ordinary skilled artisan would have been motivated to do so as taught by Kariko because these modified mRNAs have higher translational capacity than unmodified mRNAs (Abstract).
Since the instant application claims are obvious over cited patent claims in view of Gill et al., said claims are not patentably distinct.
Claims 17 and 20 are provisionally rejected on the grounds of nonstatutory double patenting as being unpatentable over claims 18-20 of copending Application No. 19/061,415), in view of Gill et al., (US2018/0244748, filed 7/28/2016, published 8/30/2018, see IDS filed 7/20/2023), in further view of Sahin et al. (WO2014/072061, filed 11/07/2013, published 5/15/2014)
As discussed previously, Gill makes obvious a method of modifying macrophages comprising transfection with modified mRNA encoding CAR
However, Gill is silent with respect to further including step of treating the cells with an RNAaseL inhibitor.
Sahir teaches methods of overexpressing IVT modified RNA in cells including macrophages (Abstract, p. 67, 1st para., see Claims 1-5 of Sahir).
In regard to claim 17, Sahin teaches the steps of inhibiting intracellular IFN signaling by inhibiting RNAseL (p. 3, 4th para., p. 49, 1st para., see Claims 12-15 and 30 of Sahir).
In regard to claim 20, Sahin teaches the cells are treated with the inhibitor prior to delivering the mRNA encoding the protein to be expressed (p. 50, 4th para.).
Accordingly, it would have been prima facie obvious to one of ordinary skill in the art at the time of filing to claim the method of modifying macrophages comprising transfection with modified mRNA encoding CAR as taught by Gill and to combine step of inhibiting RNaseL as taught by Sahin with a reasonable expectation of success. The ordinary skilled artisan would have been motivated to do so as taught by Sahin because if the intracellular IFN signaling mediated by RNaseL is not inhibited, then RNA degradation occurs and/or the inhibition of RNA translation (p. 3, 4th para., p. 49, 1st para.). In regard to choosing to treat the cells with the RNaseL inhibitor before RNA delivery, it would have been obvious to choose the pretreatment option so as to avoid mRNA degradation upon delivery.
Since the instant application claims are obvious over cited patent claims in view of Gill et al., said claims are not patentably distinct.
Claim 18 is provisionally rejected on the grounds of nonstatutory double patenting as being unpatentable over claims 18-20 of copending Application No. 19/061,415), in view of Gill et al., (US2018/0244748, filed 7/28/2016, published 8/30/2018, see IDS filed 7/20/2023) and Sahin et al. (WO2014/072061, filed 11/07/2013, published 5/15/2014, in further view of Jha et al. (JBC, 2011, 286:26319-26326)
As discussed previously, Gill in view of Sahin make obvious a method of modifying macrophages comprising transfection with modified mRNA encoding CAR and inhibiting RNaseL.
However, although Sahir teaches the inhibitor of IFN intracellular signaling is an imidazole-oxindole compound, Gill et al. are silent with respect the RNAaseL inhibitor as the oxindole compound of sinitunib.
Jha teaches that sunitinib is a potent inhibitor of RNaseL in cultured mammalian cells (Abstract, Introduction).
Accordingly, it would have been prima facie obvious to one of ordinary skill in the art at the time of filing to claim the method of modifying macrophages with a CAR mRNA and further combine an RNaseL inhibitor to improve mRNA expression as suggested by Gill and Sahin and choose sunitinib as the RNaseL inhibitor as taught by Jha with a reasonable expectation of success. The ordinary skilled artisan would have been motivated to do so as taught by Jha because of the in vitro potency of sunitinib in cultured mammalian cells.
Since the instant application claims are obvious over cited patent claims in view of Gill et al., said claims are not patentably distinct.
Claims 21-24 are is provisionally rejected on the grounds of nonstatutory double patenting as being unpatentable over claims 18-20 of copending Application No. 19/061,415), in view of Gill et al., (US2018/0244748, filed 7/28/2016, published 8/30/2018, see IDS filed 7/20/2023), in view of Xie et al. (IJMM, 2016, 38:148-160)
As discussed previously, Gill make obvious a method of modifying macrophages comprising transfection with modified mRNA encoding CAR.
However, although Gill suggests the method steps of stimulating the CAR monocytes (i.e., CARMA) by treatments with synergistic compounds such as cytokines after delivering of the CAR nucleic acid ([0127, 0384], Fig. 28), and Gill discloses the ability of macrophages to become polarized, and teaches the anti-tumor advantages of up-regulation of M1 markers in inflammatory/activated macrophages [0012, 0283, 0374], see also Claims 1 and 9 of Gill), Gill does not reduce to practice the step of culturing the RNA transfected macrophages with IFN-beta.
Xie teaches methods of treating macrophages with cytokines in order to induce M1 polarization and teaches that media comprising IFN-beta is required for sustained M1 polarization (Abstract a chimeric receptors (Abstract).
Accordingly, it would have been prima facie obvious to one of ordinary skill in the art at the time of filing to claim the method of modifying macrophages comprising transfection with modified mRNA encoding CAR and to treat the CARMA with cytokines to induce a M1 phenotype as suggested by Gill and to combine IFN-beta in the cytokine treatment as suggested by Xie with a reasonable expectation of success. The ordinary skilled artisan would have been motivated to do so as taught by Xie because IFN-beta plays a crucial role in the regulation of M1 polarization of macrophages (Abstract, Discussion, p. 158).
Since the instant application claims are obvious over cited patent claims in view of Gill et al., said claims are not patentably distinct.
Claims 1, 2, 5, 6, 21-22, and 25-27 are provisionally rejected on the grounds of nonstatutory double patenting as being unpatentable over claims 24 and 25 of copending Application No. 18/809,915, in view of Gill et al., (US2018/0244748, filed 7/28/2016, published 8/30/2018, see IDS filed 7/20/2023). This is a provisional nonstatutory double patenting rejection because the patentably indistinct claims have not in fact been patented
The subject matter claimed in the instant application is disclosed in the referenced application as follows: the method for modifying an immune cell comprising introducing a nucleic acid encoding a CAR makes obvious the method of instant application. It is clear that elements of the cited application claims are to be found in instant claims. The difference between the cited application claims and the instant claims lies in the fact that the instant application claims are more specific because they claim a modified and purified mRNA.
In regard to claim 1, Gill teaches an in vitro method of modifying human macrophages comprising the following steps:
Modifying a mRNA encoding a CAR produced by a mMessage mMachine T7 ultras In Vitro Transcription kit,
Purifying the mRNA using a Qiagen RNEasy RNA purification kit, and
Delivering the mRNA into the macrophages by electroporation ([0325], Figs 12 & 13, see also Claims 18-20 of Gill).
In regard to claim 2, Gill teaches the mRNA is modified have both a cap on the 5’end and a 3’ poly(A) tail which determine ribosome binding, initiation of translation and stability of the mRNA [0267, 0272].
Accordingly it would have been obvious to claim the cited method and to further claim a purified modified mRNA as taught by Gill with a reasonable expectation of success. One of ordinary skill in the art would have claimed so because of the high efficiency of CAR transfection with purified modified RNA.
In regard to claim 5, as stated supra, Gill teaches the purification step using the RNA using a Qiagen RNEasy RNA purification kit, which comprises a silica membrane purification column.
In regard to claim 6, as stated supra, Gill teaches transfection by electroporation in order to achieve high efficiency transfection.
In regard to claims 21, 22, and 25, Gill teaches the CAR macrophages are cultured for 30 min with beta-glucan and then tested in an in vitro killing assay [0326], which would have been obvious to claim so as to test the efficacy of the CARMA.
In regard to claim 25, as stated supra, Gill teaches the macrophages express the CAR.
In regard to claims 26 and 27, as stated supra, Gill teaches the mRNA is modified with a 5’cap and a 3’ poly(A) tail which contributed to stability of the mRNA.
Since the instant application claims are obvious over cited application claims, in view of Gill, said claims are not patentably distinct.
Claim 3 is provisionally rejected on the grounds of nonstatutory double patenting as being unpatentable over claims 24 and 25 of copending Application No. 18/809,915), in view of Gill et al., (US2018/0244748, filed 7/28/2016, published 8/30/2018, see IDS filed 7/20/2023), in view further of Grudzien-Nogalska (Meth Enzym, 2007, 431:203-227)
As discussed previously, Gill makes obvious an in vitro method of modifying macrophages comprising transfection with modified mRNA encoding a CAR.
However, although Gill makes obvious the mRNA is modified such as with a 5’ cap, they are silent to an ARAC 5’ cap.
In regard to claim 3, Grudzien-Nogalska reviews ARACs and their applications in mRNA translation and stability (p. 204-224).
Accordingly, it would have been prima facie obvious to one of ordinary skill in the art at the time of filing to claim the method of modifying macrophages comprising transfection with modified CAR mRNA comprising a 5’cap as taught by Gill and to choose the ARAC as taught by Gurdzien-Nogalska with a reasonable expectation of success. The ordinary skilled artisan would have been motivated to do so as taught by Gurdzien-Nogalska because ARACs show two-fold higher translational efficiency compared to conventional caps (p. 205, Introduction).
Since the instant application claims are obvious over cited patent claims in view of Gill et al., said claims are not patentably distinct.
Claim 4 is provisionally rejected on the grounds of nonstatutory double patenting as being unpatentable over claims 24 and 25 of copending Application No. 18/809,915), in view of Gill et al., (US2018/0244748, filed 7/28/2016, published 8/30/2018, see IDS filed 7/20/2023), in further view of Kariko et al. (Mol Ther, 2008, 16:1833-1840)
As discussed previously, Gill makes a method of modifying macrophages comprising transfection with modified mRNA encoding a CAR.
However, although Gill suggest using nucleotide analogues [0265], they are silent to the mRNA being modified with a pseudouridine.
In regard to claim 4, Kariko teaches methods of expressing mRNA in cells, wherein the mRNA is modified to include psuedouridine (Abstract).
Accordingly, it would have been prima facie obvious to one of ordinary skill in the art at the time of filing to claim the method of modifying macrophages comprising transfection with modified CAR mRNA as taught by Gill and substitute modified nucleosides such as a pseudouridine as taught by Kariko with a reasonable expectation of success. The ordinary skilled artisan would have been motivated to do so as taught by Kariko because these modified mRNAs have higher translational capacity than unmodified mRNAs (Abstract).
Since the instant application claims are obvious over cited patent claims in view of Gill et al., said claims are not patentably distinct.
Claims 17 and 20 are provisionally rejected on the grounds of nonstatutory double patenting as being unpatentable over claims 24 and 25 of copending Application No. 18/809,915), in view of Gill et al., (US2018/0244748, filed 7/28/2016, published 8/30/2018, see IDS filed 7/20/2023), in further view of Sahin et al. (WO2014/072061, filed 11/07/2013, published 5/15/2014)
As discussed previously, Gill makes obvious a method of modifying macrophages comprising transfection with modified mRNA encoding CAR
However, Gill is silent with respect to further including step of treating the cells with an RNAaseL inhibitor.
Sahir teaches methods of overexpressing IVT modified RNA in cells including macrophages (Abstract, p. 67, 1st para., see Claims 1-5 of Sahir).
In regard to claim 17, Sahin teaches the steps of inhibiting intracellular IFN signaling by inhibiting RNAseL (p. 3, 4th para., p. 49, 1st para., see Claims 12-15 and 30 of Sahir).
In regard to claim 20, Sahin teaches the cells are treated with the inhibitor prior to delivering the mRNA encoding the protein to be expressed (p. 50, 4th para.).
Accordingly, it would have been prima facie obvious to one of ordinary skill in the art at the time of filing to claim the method of modifying macrophages comprising transfection with modified mRNA encoding CAR as taught by Gill and to combine step of inhibiting RNaseL as taught by Sahin with a reasonable expectation of success. The ordinary skilled artisan would have been motivated to do so as taught by Sahin because if the intracellular IFN signaling mediated by RNaseL is not inhibited, then RNA degradation occurs and/or the inhibition of RNA translation (p. 3, 4th para., p. 49, 1st para.). In regard to choosing to treat the cells with the RNaseL inhibitor before RNA delivery, it would have been obvious to choose the pretreatment option so as to avoid mRNA degradation upon delivery.
Since the instant application claims are obvious over cited patent claims in view of Gill et al., said claims are not patentably distinct.
Claim 18 is provisionally rejected on the grounds of nonstatutory double patenting as being unpatentable over claims 24 and 25 of copending Application No. 18/809,915), in view of Gill et al., (US2018/0244748, filed 7/28/2016, published 8/30/2018, see IDS filed 7/20/2023) and Sahin et al. (WO2014/072061, filed 11/07/2013, published 5/15/2014, in further view of Jha et al. (JBC, 2011, 286:26319-26326)
As discussed previously, Gill in view of Sahin make obvious a method of modifying macrophages comprising transfection with modified mRNA encoding CAR and inhibiting RNaseL.
However, although Sahir teaches the inhibitor of IFN intracellular signaling is an imidazole-oxindole compound, Gill et al. are silent with respect the RNAaseL inhibitor as the oxindole compound of sinitunib.
Jha teaches that sunitinib is a potent inhibitor of RNaseL in cultured mammalian cells (Abstract, Introduction).
Accordingly, it would have been prima facie obvious to one of ordinary skill in the art at the time of filing to claim the method of modifying macrophages with a CAR mRNA and further combine an RNaseL inhibitor to improve mRNA expression as suggested by Gill and Sahin and choose sunitinib as the RNaseL inhibitor as taught by Jha with a reasonable expectation of success. The ordinary skilled artisan would have been motivated to do so as taught by Jha because of the in vitro potency of sunitinib in cultured mammalian cells.
Since the instant application claims are obvious over cited patent claims in view of Gill et al., said claims are not patentably distinct.
Claims 21-24 are provisionally rejected on the grounds of nonstatutory double patenting as being unpatentable over claims 24 and 25 of copending Application No. 18/809,915), in view of Gill et al., (US2018/0244748, filed 7/28/2016, published 8/30/2018, see IDS filed 7/20/2023), in view of Xie et al. (IJMM, 2016, 38:148-160)
As discussed previously, Gill make obvious a method of modifying macrophages comprising transfection with modified mRNA encoding CAR.
However, although Gill suggests the method steps of stimulating the CAR monocytes (i.e., CARMA) by treatments with synergistic compounds such as cytokines after delivering of the CAR nucleic acid ([0127, 0384], Fig. 28), and Gill discloses the ability of macrophages to become polarized, and teaches the anti-tumor advantages of up-regulation of M1 markers in inflammatory/activated macrophages [0012, 0283, 0374], see also Claims 1 and 9 of Gill), Gill does not reduce to practice the step of culturing the RNA transfected macrophages with IFN-beta.
Xie teaches methods of treating macrophages with cytokines in order to induce M1 polarization and teaches that media comprising IFN-beta is required for sustained M1 polarization (Abstract a chimeric receptors (Abstract).
Accordingly, it would have been prima facie obvious to one of ordinary skill in the art at the time of filing to claim the method of modifying macrophages comprising transfection with modified mRNA encoding CAR and to treat the CARMA with cytokines to induce a M1 phenotype as suggested by Gill and to combine IFN-beta in the cytokine treatment as suggested by Xie with a reasonable expectation of success. The ordinary skilled artisan would have been motivated to do so as taught by Xie because IFN-beta plays a crucial role in the regulation of M1 polarization of macrophages (Abstract, Discussion, p. 158).
Since the instant application claims are obvious over cited patent claims in view of Gill et al., said claims are not patentably distinct.
Claims 1, 2, 5, 6, 21-22, and 25-27 are provisionally rejected on the grounds of nonstatutory double patenting as being unpatentable over claims 96 and 101 of copending Application No. 17/477,491, in view of Gill et al., (US2018/0244748, filed 7/28/2016, published 8/30/2018, see IDS filed 7/20/2023). This is a provisional nonstatutory double patenting rejection because the patentably indistinct claims have not in fact been patented
The subject matter claimed in the instant application is disclosed in the referenced application as follows: the method for modifying an immune cell comprising introducing a nucleic acid encoding a CAR makes obvious the method of instant application. It is clear that elements of the cited application claims are to be found in instant claims. The difference between the cited application claims and the instant claims lies in the fact that the instant application claims are more specific because they claim a modified and purified mRNA.
In regard to claim 1, Gill teaches an in vitro method of modifying human macrophages comprising the following steps:
Modifying a mRNA encoding a CAR produced by a mMessage mMachine T7 ultras In Vitro Transcription kit,
Purifying the mRNA using a Qiagen RNEasy RNA purification kit, and
Delivering the mRNA into the macrophages by electroporation ([0325], Figs 12 & 13, see also Claims 18-20 of Gill).
In regard to claim 2, Gill teaches the mRNA is modified have both a cap on the 5’end and a 3’ poly(A) tail which determine ribosome binding, initiation of translation and stability of the mRNA [0267, 0272].
Accordingly it would have been obvious to claim the cited method and to further claim a purified modified mRNA as taught by Gill with a reasonable expectation of success. One of ordinary skill in the art would have claimed so because of the high efficiency of CAR transfection with purified modified RNA.
In regard to claim 5, as stated supra, Gill teaches the purification step using the RNA using a Qiagen RNEasy RNA purification kit, which comprises a silica membrane purification column.
In regard to claim 6, as stated supra, Gill teaches transfection by electroporation in order to achieve high efficiency transfection.
In regard to claims 21, 22, and 25, Gill teaches the CAR macrophages are cultured for 30 min with beta-glucan and then tested in an in vitro killing assay [0326], which would have been obvious to claim so as to test the efficacy of the CARMA.
In regard to claim 25, as stated supra, Gill teaches the macrophages express the CAR.
In regard to claims 26 and 27, as stated supra, Gill teaches the mRNA is modified with a 5’cap and a 3’ poly(A) tail which contributed to stability of the mRNA.
Since the instant application claims are obvious over cited application claims, in view of Gill, said claims are not patentably distinct.
Claim 3 is provisionally rejected on the grounds of nonstatutory double patenting as being unpatentable over claims 96 and 101 of copending Application No. 17/477,491), in view of Gill et al., (US2018/0244748, filed 7/28/2016, published 8/30/2018, see IDS filed 7/20/2023), in view further of Grudzien-Nogalska (Meth Enzym, 2007, 431:203-227)
As discussed previously, Gill makes obvious an in vitro method of modifying macrophages comprising transfection with modified mRNA encoding a CAR.
However, although Gill makes obvious the mRNA is modified such as with a 5’ cap, they are silent to an ARAC 5’ cap.
In regard to claim 3, Grudzien-Nogalska reviews ARACs and their applications in mRNA translation and stability (p. 204-224).
Accordingly, it would have been prima facie obvious to one of ordinary skill in the art at the time of filing to claim the method of modifying macrophages comprising transfection with modified CAR mRNA comprising a 5’cap as taught by Gill and to choose the ARAC as taught by Gurdzien-Nogalska with a reasonable expectation of success. The ordinary skilled artisan would have been motivated to do so as taught by Gurdzien-Nogalska because ARACs show two-fold higher translational efficiency compared to conventional caps (p. 205, Introduction).
Since the instant application claims are obvious over cited patent claims in view of Gill et al., said claims are not patentably distinct.
Claim 4 is provisionally rejected on the grounds of nonstatutory double patenting as being unpatentable over claims 96 and 101 of copending Application No. 17/477,491), in view of Gill et al., (US2018/0244748, filed 7/28/2016, published 8/30/2018, see IDS filed 7/20/2023), in further view of Kariko et al. (Mol Ther, 2008, 16:1833-1840)
As discussed previously, Gill makes a method of modifying macrophages comprising transfection with modified mRNA encoding a CAR.
However, although Gill suggest using nucleotide analogues [0265], they are silent to the mRNA being modified with a pseudouridine.
In regard to claim 4, Kariko teaches methods of expressing mRNA in cells, wherein the mRNA is modified to include psuedouridine (Abstract).
Accordingly, it would have been prima facie obvious to one of ordinary skill in the art at the time of filing to claim the method of modifying macrophages comprising transfection with modified CAR mRNA as taught by Gill and substitute modified nucleosides such as a pseudouridine as taught by Kariko with a reasonable expectation of success. The ordinary skilled artisan would have been motivated to do so as taught by Kariko because these modified mRNAs have higher translational capacity than unmodified mRNAs (Abstract).
Since the instant application claims are obvious over cited patent claims in view of Gill et al., said claims are not patentably distinct.
Claims 17 and 20 are provisionally rejected on the grounds of nonstatutory double patenting as being unpatentable over claims 96 and 101 of copending Application No. 17/477,491), in view of Gill et al., (US2018/0244748, filed 7/28/2016, published 8/30/2018, see IDS filed 7/20/2023), in further view of Sahin et al. (WO2014/072061, filed 11/07/2013, published 5/15/2014)
As discussed previously, Gill makes obvious a method of modifying macrophages comprising transfection with modified mRNA encoding CAR
However, Gill is silent with respect to further including step of treating the cells with an RNAaseL inhibitor.
Sahir teaches methods of overexpressing IVT modified RNA in cells including macrophages (Abstract, p. 67, 1st para., see Claims 1-5 of Sahir).
In regard to claim 17, Sahin teaches the steps of inhibiting intracellular IFN signaling by inhibiting RNAseL (p. 3, 4th para., p. 49, 1st para., see Claims 12-15 and 30 of Sahir).
In regard to claim 20, Sahin teaches the cells are treated with the inhibitor prior to delivering the mRNA encoding the protein to be expressed (p. 50, 4th para.).
Accordingly, it would have been prima facie obvious to one of ordinary skill in the art at the time of filing to claim the method of modifying macrophages comprising transfection with modified mRNA encoding CAR as taught by Gill and to combine step of inhibiting RNaseL as taught by Sahin with a reasonable expectation of success. The ordinary skilled artisan would have been motivated to do so as taught by Sahin because if the intracellular IFN signaling mediated by RNaseL is not inhibited, then RNA degradation occurs and/or the inhibition of RNA translation (p. 3, 4th para., p. 49, 1st para.). In regard to choosing to treat the cells with the RNaseL inhibitor before RNA delivery, it would have been obvious to choose the pretreatment option so as to avoid mRNA degradation upon delivery.
Since the instant application claims are obvious over cited patent claims in view of Gill et al., said claims are not patentably distinct.
Claim 18 is provisionally rejected on the grounds of nonstatutory double patenting as being unpatentable over claims 96 and 101 of copending Application No. 17/477,491), in view of Gill et al., (US2018/0244748, filed 7/28/2016, published 8/30/2018, see IDS filed 7/20/2023) and Sahin et al. (WO2014/072061, filed 11/07/2013, published 5/15/2014, in further view of Jha et al. (JBC, 2011, 286:26319-26326)
As discussed previously, Gill in view of Sahin make obvious a method of modifying macrophages comprising transfection with modified mRNA encoding CAR and inhibiting RNaseL.
However, although Sahir teaches the inhibitor of IFN intracellular signaling is an imidazole-oxindole compound, Gill et al. are silent with respect the RNAaseL inhibitor as the oxindole compound of sinitunib.
Jha teaches that sunitinib is a potent inhibitor of RNaseL in cultured mammalian cells (Abstract, Introduction).
Accordingly, it would have been prima facie obvious to one of ordinary skill in the art at the time of filing to claim the method of modifying macrophages with a CAR mRNA and further combine an RNaseL inhibitor to improve mRNA expression as suggested by Gill and Sahin and choose sunitinib as the RNaseL inhibitor as taught by Jha with a reasonable expectation of success. The ordinary skilled artisan would have been motivated to do so as taught by Jha because of the in vitro potency of sunitinib in cultured mammalian cells.
Since the instant application claims are obvious over cited patent claims in view of Gill et al., said claims are not patentably distinct.
Claims 21-24 are provisionally rejected on the grounds of nonstatutory double patenting as being unpatentable over claims 96 and 101 of copending Application No. 17/477,491), in view of Gill et al., (US2018/0244748, filed 7/28/2016, published 8/30/2018, see IDS filed 7/20/2023), in view of Xie et al. (IJMM, 2016, 38:148-160)
As discussed previously, Gill make obvious a method of modifying macrophages comprising transfection with modified mRNA encoding CAR.
However, although Gill suggests the method steps of stimulating the CAR monocytes (i.e., CARMA) by treatments with synergistic compounds such as cytokines after delivering of the CAR nucleic acid ([0127, 0384], Fig. 28), and Gill discloses the ability of macrophages to become polarized, and teaches the anti-tumor advantages of up-regulation of M1 markers in inflammatory/activated macrophages [0012, 0283, 0374], see also Claims 1 and 9 of Gill), Gill does not reduce to practice the step of culturing the RNA transfected macrophages with IFN-beta.
Xie teaches methods of treating macrophages with cytokines in order to induce M1 polarization and teaches that media comprising IFN-beta is required for sustained M1 polarization (Abstract a chimeric receptors (Abstract).
Accordingly, it would have been prima facie obvious to one of ordinary skill in the art at the time of filing to claim the method of modifying macrophages comprising transfection with modified mRNA encoding CAR and to treat the CARMA with cytokines to induce a M1 phenotype as suggested by Gill and to combine IFN-beta in the cytokine treatment as suggested by Xie with a reasonable expectation of success. The ordinary skilled artisan would have been motivated to do so as taught by Xie because IFN-beta plays a crucial role in the regulation of M1 polarization of macrophages (Abstract, Discussion, p. 158).
Since the instant application claims are obvious over cited patent claims in view of Gill et al., said claims are not patentably distinct.
Conclusion
No claims are allowed.
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/ARTHUR S LEONARD/Examiner, Art Unit 1631