DETAILED ACTION
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 .
Status of Application/Amendment/Claims
This Office action is in response to the communications filed on February 2, 2026.
Currently, claims 1-4, 8, 18-22, 24-30, and 33 are pending and under examination on the merits in the instant application.
The following rejections are either newly applied or are reiterated and are the only rejections and/or objections presently applied to the instant application.
Response to Arguments and Amendments
Withdrawn Rejections
Any rejections/objections not repeated in this Office action are hereby withdrawn.
Terminal Disclaimer
The terminal disclaimer filed on February 2, 2026 disclaiming the terminal portion of any patent granted on this application which would extend beyond the expiration date of Application No. 18/794,415 has been reviewed and is accepted. The terminal disclaimer has been recorded.
New Objections/Rejections Necessitated by Amendment
Priority
Applicant’s claim for the benefit of a prior-filed application under 35 U.S.C. 119(e) or under 35 U.S.C. 120, 121, 365(c), or 386(c) is acknowledged. Applicant has not complied with one or more conditions for receiving the benefit of an earlier filing date under 35 U.S.C. 119(e) or under 35 U.S.C. 120, 121, 365(c), or 386(c) as follows:
The later-filed application must be an application for a patent for an invention which is also disclosed in the prior application (the parent or original nonprovisional application or provisional application). The disclosure of the invention in the parent application and in the later-filed application must be sufficient to comply with the requirements of 35 U.S.C. 112(a) or the first paragraph of pre-AIA 35 U.S.C. 112, except for the best mode requirement. See Transco Products, Inc. v. Performance Contracting, Inc., 38 F.3d 551, 32 USPQ2d 1077 (Fed. Cir. 1994).
The disclosure of the prior-filed applications, Application Nos. 63/437,855, 18/708,093, and 18/794,415, fails to provide adequate support or enablement in the manner provided by 35 U.S.C. 112(a) or pre-AIA 35 U.S.C. 112, first paragraph for one or more claims of this application. It is noted that none of the prior-filed applications describes the newly introduced limitation pertaining to the phosphorothioate positions, LNA modified base positions, and the mixture of 2’-O-methyl and LNA bases. Accordingly, claims 1-4, 8, 18-22, 24-30, and 33 are not entitled to the benefit of an earlier filing date. Therefore, the effective filing date for claims 1-4, 8, 18-22, 24-30, and 33 will be the filing date of the instant application, which is October 31, 2024.
Claim Rejections - 35 USC § 112
The text of those sections of Title 35, U.S. Code not included in this action can be found in a prior Office action.
Claims 1-4, 8, 18-22, 24-30, and 33 are rejected under 35 U.S.C. 112(b) or 35 U.S.C. 112 (pre-AIA ), second paragraph, as being indefinite for failing to particularly point out and distinctly claim the subject matter which the inventor or a joint inventor (or for applications subject to pre-AIA 35 U.S.C. 112, the applicant), regards as the invention.
Claim 1 and all of dependent claims thereof now recite “the oligonucleotide comprises a sequence of 2’-O-methyl modified bases and locked nucleic acid (LNA) modified bases, wherein the LNA modified bases are positioned at approximately every third nucleotide within the sequence.”. The claims fail to particularly point out and distinctly claim the positions for the “2’-O-methyl modified bases” within the “sequence”. As such, it is unclear whether the oligonucleotide sequence is required to comprise only two 2’-O-methyl modified bases at any positions within the sequence (see the plural term “bases”) or is required to comprise more than two 2’-O-methyl modified bases at certain positions within the sequence. Therefore, the chemical modification limitations pertaining to the oligonucleotide cannot be clearly ascertained, thereby rendering the claimed structure indefinite.
Claim 1 and all of dependent claims thereof now recite “the LNA modified bases are positioned at approximately every third nucleotide within the sequence.”
The term “approximately” is a relative term which renders the claim indefinite. The term “approximately” is not defined by the claim, nor is the term clearly defined in the specification. As such, one of ordinary skill in the art would not reasonably ascertain the clear metes and bounds pertaining to “approximately every third” as the term “approximately” can be defined differently by different persons of ordinary skill in the art, thereby rendering the claims indefinite.
Claim Rejections - 35 USC § 112
The text of those sections of Title 35, U.S. Code not included in this action can be found in a prior Office action.
Claims 1-4, 8, 18-22, 24-30, and 33 are rejected under 35 U.S.C. 112(a) or 35 U.S.C. 112 (pre-AIA ), first paragraph, as failing to comply with the written description requirement. The claim(s) contains subject matter which was not described in the specification in such a way as to reasonably convey to one skilled in the relevant art that the inventor or a joint inventor, or for applications subject to pre-AIA 35 U.S.C. 112, the inventor(s), at the time the application was filed, had possession of the claimed invention. This includes a new matter rejection.
The claims as currently amended now require the following structural requirements regarding the miR-193b-5p inhibitor:
1) “a synthetic single-stranded oligonucleotide comprising phosphorothioate linkages at its 3’ and 5’ ends”;
2) “the oligonucleotide comprises a sequence of 2’-O-methyl modified bases and locked nucleic acid (LNA) modified bases”; and
3) “the LNA modified bases are positioned at approximately every third nucleotide within the sequence.”
Nowhere in the instant specification is there a description or a disclosure of a synthetic single-stranded oligonucleotide sequence that inhibits miR-193b-5p, wherein the oligonucleotide sequence comprises a phosphorothioate linkage at each of the 5’ end and the 3’ end and a mixture of both 2’-O-methyl bases and LNA bases, wherein the LNA bases are positioned “approximately every third nucleotide”, wherein the “approximately every third nucleotide” reads on every nucleotide, every second nucleotide, every third nucleotide, every fourth nucleotide, and every fifth nucleotide in light of the relative term “approximately”, which is ambiguous as set forth in the §112(b) rejection above.
Now, as applicant must be aware, the instant specification is completely silent regarding any nucleotide sequence, let alone the instantly claimed “a sequence of 2’-O-methyl modified bases and locked nucleic acid (LNA) modified bases” as evidenced by the lack of a sequence listing in the instant application. In addition to the objective fact that there is no actually disclosed oligonucleotide sequence representing the instantly claimed miR-193b-5p inhibitor that is now claimed to have specific chemical modifications at specific positions, the instant specification is completely devoid of any written description support for the instantly claimed method of treating sepsis in a subject comprising administering the oligonucleotide “sequence” that is specifically chemically modified as currently claimed. Note that it was art-recognized knowledge that the miRNA inhibitor activity greatly varies depending on the specific nucleotide sequence (also length) and the specific chemical modifications. See for instance Table C of Bhat et al. (US 9,157,083 B2) demonstrating highly sequence/length/chemical modification-dependent anti-miR-122 compound activity/potency, wherein the compound # 38021 comprises LNA at positions 1, 4, 7, 8, 10, 12, 14, and 15 and a 2’-MOE at position 16. As such, in view of the unpredictable nature of miRNA inhibitor oligonucleotides having different sequences/lengths with different modification patterns, the required structure-function correlation for the instantly claimed genus encompassing any oligonucleotide sequence/length with various modifications cannot be deemed adequately described by the instant specification, which is completely devoid of any nucleotide sequence, let alone a chemically modified sequence targeting miR-193b-5p.
Accordingly, the instant specification fails to reasonably convey that the instant co-inventors had possession of the instantly claimed sepsis treatment method that requires administration of a specifically modified oligonucleotide sequence, which is not disclosed in the instant application, as of the filing date sought or granted in the instant application.
It is very interesting to note that applicant expressly pointed out “original claims 16 and 17” of the instant application for LNA modified bases in the remarks filed on February 2, 2026. In response, it is noted that there is no description/limitation in original claims 16-17 or anywhere in the instant application supporting the instantly claimed “wherein the LNA modified bases are positioned at approximately every third nucleotide within the sequence.” Furthermore, there is no written description support for the “phosphorothioate linkages at its 3’ and 5’ ends” and the combination of “2’-O-methyl modified bases and locked nucleic acid (LNA) modified bases”, wherein the position/number of LNA modifications would depend on the actual length (not disclosed) of the oligonucleotide.
Accordingly, claims 1-4, 8, 18-22, 24-30, and 33 introduce new matter that is not described in the instant specification and original claims as originally filed.
Claim Rejections - 35 USC § 103
Claims 1-2, 4, and 31 are rejected under 35 U.S.C. 103 as being unpatentable over dos Santos et al. (European Respiratory Journal, published on January 6, 2022, 59:2004216, of record) in view of Fourie (US 2006/0111398 A1, of record), Thompson et al. (US 2014/0371299 A1, of record), Mazzone et al. (US 2021/0087554 A1, of record), and Lennox et al. (Molecular Therapy – Nucleic Acids, 2013, 2:e117).
Solely in the interest of compact prosecution, the word “approximately” will not be taken into consideration. See the §112(b) and §112(a) rejections above.
dos Santos teaches a method comprising administering a miR-193b-5p inhibitor (“INH”) in “lipid nanoparticles” to mice challenged with LPS and observing “attenuated” LPS-induced lung injury in the mice, wherein the inhibitor is “commercially available” and “purchased” and “is modified to reduce in vivo degradation (2’-O-methyl-group-modified oligonucleotides).” See pages 8 and 13; Figures 5a)-5l).
dos Santos teaches that the studies demonstrate “the impact of miR-193b-5p on lung inflammation” and that one can “explore delivering the microRNA inhibitor later in the course of injury, and for longer pre-clinical models, to address the ability of the [miR-193b-5p] inhibitor to either limit or even reverse pulmonary oedema once the cascade leading to loss of vascular integrity is initiated.” See pages 2 and 13.
dos Santos reports that the overall survival % in mice challenged with LPS inversely correlates with the expression/presence of miR-193b-5p such that mice lacking miR-193b-5p has a significantly higher survival % compared to mice heterozygous for miR-193b-5p or the wild-type mice. See Figure 4a).
dos Santos reports that miR-193b-5p expression is significantly increased in lungs from patients with acute respiratory distress syndrome (ARDS) with diffuse alveolar damage (DAD) comped to ARDS patients without DAD. See page 8; Figure 7a). Hence, dos Santos teaches that increased expression level of miR-193b-5p has “clinical relevance” in sepsis-induced ARDS and that miR-196b-5p is a “therapeutic target for sepsis-induced ALI.” See pages 2 and 8.
dos Santos teaches that “sepsis is the leading cause of acute respiratory distress syndrome (ARDS), the clinical syndrome of acute lung injury (ALI) in humans.” See page 1.
dos Santos does not expressly disclose that the mice treated with LPS is an art-recognized animal model of sepsis.
dos Santos does not teach observing extended survival or a higher survival % and reduced organ dysfunction in the miR-193b-5p inhibitor-treated mice having LPS-induced lung injury.
dos Santos does not teach that the miR-193b-5p inhibitor that is “2’-O-methyl-group-modified” is further modified with phosphorothioate linkages and LNAs every third position.
Fourie discloses, “LPS administration to animals serves as a model for LPS-induced sepsis” thus potential therapeutic agents that “improve survival of mice in LPS-induced sepsis model” have been studied in the art. See paragraphs 0005-0006.
Fourie teaches that LPS-induced inflammation includes “multiple organ dysfunction syndrome,” “acute respiratory distress syndrome,” and “neuroinflammation”. See paragraph 0154.
Thompson teaches that mice given LPS is an art-recognized septic mice models for investigating an agent that can be used to treat sepsis in a mammal. See paragraph 0110 disclosing “FIG. 19 shows the construct of the siRNA used in the septic mice models. In this study, the mice were given a dose of LPS that induces sepsis and death in the animal within 48 hours after the LPS is administered.”
Thompson teaches that “LPS is a macromolecular cell surface antigen of bacteria that when applied in vivo triggers a network of inflammatory responses.” See paragraph 0092.
Thompson teaches that sepsis “is also known as systemic inflammatory response syndrome (“SIRS”)” and that sepsis “can be simply defined as a spectrum of clinical conditions caused by the immune response of a patient to infection” thus sepsis treatment is mediated by decreasing the expression of pro-inflammatory cytokines, which are known to cause neuroinflammation. See paragraphs 0011 and 0104.
Thompson teaches that antisense oligonucleotides can be chemically modified with phosphorothioate linkages and 2’-O-methyls to “enhance their resistance to nucleases”. See paragraphs 0087-0088.
Mazzone teaches that animal “models to study sepsis” include “the ones that induce sepsis by injecting an exogenous toxin, like LPS (lipopolysaccharide)”, which “can mimic the initial clinical steps of seps in humans and increase the pro-inflammatory cytokine levels”, wherein the “LPS-induced inflammation model and the CLP model are mostly used for studying sepsis”. See paragraph 0004.
Mazzone teaches that an antisense oligonucleotide targeting a miRNA (e.g., miR-210) reduces the expression of the miRNA, wherein the antisense oligonucleotide “improved the survival rate of a sepsis mice model” or “improved survival of LPS-treated mice” and also was able to “protect against organ damage” thus can be applied “for use in treatment of sepsis”. See paragraphs 0008, 0105, and 0108-0110.
Mazzone teaches that the miRNA-targeting antisense oligonucleotide can be modified with “locked nucleic acid (LNA)”, phosphorothioates, and 2’-O-modified ribonucleotides for increased resistance to nucleases. See paragraphs 0047-0049.
Mazzone exemplifies “locked nucleic acid (LNA) oligonucleotides (12 mg/kg) against miR-210 (anti-miR-210)”, wherein “anti-miR-210 treatment improved survival of LPS-treated mice”. See paragraph 0110.
Lennox teaches that an antisense oligonucleotide (“2’OMe/LNA PS”) comprising phosphorothioate linkages, 2’-O-methyl modifications, and LNA modifications at every third position (positions 2, 5, 8, 11, 14, 17, and 19) within a 21-mer sequence provides significantly increased potency including the “highest potency” in miRNA activity compared to 2’-O-methyl modified (2’OMe or 2’OMe PS) or LNA-modified (DNA/LNA PS) inhibitors. See the Table at page 5 and Figures 4a-4b. See also pages 7-8.
It would have been obvious to one of ordinary skill in the art before the effective filing date to observe, with a reasonable expectation of success, extended survival and reduced organ dysfunction in dos Santos’ LPS-treated mice that are administered with the chemically modified miR-193b-5p inhibitor in lipid nanoparticles in view of the finding that dos Santos’ miR-193b-5p inhibitor-treated mice showed “attenuated” LPS-induced lung injury, and further in view of the objective evidence that a decreased miR-193b-5p expression level in LPS-treated mice correlates with extended survival as demonstrated in dos Santos’ experimental data in Figure 4a). As such, one of ordinary skill in the art would have reasonably expected that administration of dos Santos’ chemically modified miR-193b-5p inhibitor in lipid nanoparticles to mice challenged with LPS would not only attenuate organ (e.g., lung) dysfunction but also prolong the survival of the mice.
It would have been prima facie obvious to one of ordinary skill in the art to apply dos Santos’ administration of miR-193b-5p inhibitor to a subject having sepsis for the purpose of treating sepsis and related conditions thereof because there was an express teaching that miR-193b-5p has “clinical relevance” in sepsis-induced ARDS thus miR-193b-5p was suggested as a “therapeutic target for sepsis-induced ALI” as disclosed by dos Santos, who also suggested to “explore delivering the microRNA inhibitor later in the course of injury, and for longer pre-clinical models”, and further because one of ordinary skill in the art would have reasonably deemed that the attenuated LPS-induced lung injury observed in dos Santos’ miR-193b-5p-treated mice represents a treatment effect in a sepsis animal model based on the art-recognized knowledge that LPS-induced mice are an art-recognized sepsis animal model that is widely used in the relevant art to study sepsis and related conditions thereof such as neuroinflammation and ARDS as evidenced by the teachings of Fourie, Thompson, and Mazzone.
One of ordinary skill in the art would have been motivated to replace the 2’-O-methyl-modified oligonucleotide of dos Santos with an LNA/2’-O-methyl/PS-modified oligonucleotide by incorporating phosphorothioate linkages at each of the ends as well as LNA modifications every third position and 2’-O-methyl modifications at other positions into an antisense oligonucleotide inhibitor targeting miR-193b-5p with a reasonable expectation of success in order to improve the in vivo stability and efficacy/potency of the inhibitor that is administered to a subject in vivo for sepsis treatment purpose or ARDS treatment purpose because “2’-O-methyl-group-modified oligonucleotides” were known to “reduce in vivo degradation” as taught by dos Santaos, and further because an LNA-modified anti-miRNA antisense oligonucleotide was already demonstrated in the relevant art to provided “improved survival of LPS-treated mice” in vivo as reported by Mazzone, wherein the combination of LNA, 2’-O-methyl, and PS modifications in an anti-miRNA antisense oligonucleotide (“2’OMe/LNA PS”) in which LNA occurs every third position and PS is present at each end was already known to provide “highest potency” in miRNA activity compared to a 2’-O-methyl-modified or an LNA-modified anti-miRNA oligonucleotide as evidenced by the teachings of Lennox.
In view of the foregoing, claims 1-2, 4, and 31 taken as a whole would have been prima facie obvious before the effective filing date.
Response to Arguments/Declaration
Applicant's arguments filed on February 2, 2026 have been fully considered but they are not persuasive. Applicant argues that the claimed inhibitor having a “novel arrangement” is structurally different from the prior art’s structure and also provides unexpected results that are superior. As an initial matter, applicant’s attention is directed to the fact that the newly added limitation pertaining to the alleged “novel arrangement” is taught by the cited art, Lennox, as explained in the rejection above. Since applicant’s arguments pertaining to the unexpected results pertaining to the claimed inhibitor structure strictly rely on and refer to the declaration, the examiner will address the declaration.
The declaration under 37 CFR 1.132 filed on February 2, 2026 is insufficient to overcome the obviousness rejection under §103 because of the following reasons:
The declarants state that they conducted experiments that “compared three distinct inhibitor designs”, wherein the alleged “innovative “NM2-LNA” design” provided “~95-100%” knockdown of miR-193b-5p in BEAS-2b cells, wherein such knockdown levels are surprising and unexpected compared to the knockdown levels provided by the commercially available product and the customized product that does not comprise LNA modified bases. The declarants point out “Exhibit A” as evidence, which shows data that they obtained “prior to the filing of the provisional application”. In response, applicant’s attention is directed to the fact that the instantly rejected claims are not directed to a composition per se. The rejected claims in the instant application are drawn to a method of treating sepsis in a subject in need thereof. As such, the greater knockdown level in cells in vitro provided by the alleged “innovative “NM2-LNA” design”, which was allegedly tested before the filing date of a provisional application, is not commensurate in scope with the instantly claimed in vivo sepsis patient treatment method. Now, the examiner wishes to make clear on the record that the provisional application, Application No. 63/437,855, is completely silent regarding the alleged “innovative” miR-193b-5p design, wherein the ‘855 provisional application does not even disclose the actual nucleotide sequence with the instantly claimed chemical modifications. As such, the examiner is bewildered by the declarants’ explicit statement that the “innovative “NM2-LNA” design” was made and tested in cells in vitro prior to the ‘855 provisional application filing date when the ‘855 provisional application itself contains nothing pertaining to an oligonucleotide sequence or oligonucleotide design comprising terminal phosphorothioates and a mixture of 2’-O-methyl and LNA modifications, wherein the LNA modifications are required to occur every third position.
Even better, it has long been expected in the prior art that a single-stranded anti-miRNA antisense oligonucleotide comprising a mixture of LNA, 2’-O-methyl, and phosphorothioate modifications performs the best by providing the most potent target miRNA inhibition in cells in vitro compared to those comprising only 2’-O-methyl and phosphorothioate modifications as evidenced by Torres et al. (Artificial DNA: PNA & XNA, 2011, 2:71-78). See Figure 3B, which demonstrates that “LNA/OMe PS” shows more than 2-fold increase in miR-122 inhibition compared to “OMe PS” in cells up to 96 hours after incubation without any transfection agents. See also page 75 disclosing the following: “The highest potency anti-miR was found to be the LNA/OMe PS ON.” (emphasis added). See also page 76 that teaches “the potent and long-lasting gymnotic anti-miR activity at sub-micromolar concentrations in cells suggests that LNA/OMe PS ONs should be given greater consideration for in vivo use.” (emphasis added). Consistent with Torres’ findings, Lennox et al. (Molecular Therapy – Nucleic Acids, 2013, 2:e117) also demonstrate that “2’OMe/LNA PS” antisense oligonucleotide targeting a miRNA provides 2-3-fold improvement in miRNA inhibition activity compared to 2’OMe oligonucleotide or 2’OMePS oligonucleotide in cells after 6 hour and 24 hours transfection. See Figures 4a-4b. See also page 7 reporting the following: “The unmodified “2’OMe” AMO showed low activity, most likely due to exonuclease degradation; the fully PS-modified version (“2’OMe PS”) also showed low activity”, wherein the “original antagomir design, 2’OMe with only three PS linkages on each end, showed improved activity.” See also page 8 reporting the following: “The 2’OMe/LNA chimera is a less commonly use AMO design, but was shown to have very high potency in earlier studies. Indeed, this design showed the highest potency of any of the miR-21 AMOs tested…the original antagomir design (“miR-24 2’OMe 3PSends”) had the lowest potency…the 2’OMe/LNA chimera (“miR-24 2’OMe/LNA PS”) showed similar increases in potency.” (emphasis added). As such, the §1.132 declaration pertaining to in vitro cell transfection experiments is far from supporting the alleged surprising, unexpected results, which are in fact merely expected, even if the rejected claims were to be drawn to an in vitro method of reducing miR-193b-5p expression in cells in view of the prior art knowledge as evidenced by Torres and Lennox.
In addition, it is noted that “Exhibit A” discloses the sequence and chemical modifications of “NM2-LNA”, which is nowhere to be found in any of the earlier-filed applications or in the instant application, wherein “NM2-LNA” comprises three consecutive phosphorothioate linkages at each of the 5’ end and the 3’ end, wherein LNA is at positions 6, 9, 12, 15, and 18 of the 21-mer sequence, and wherein 2’-O-methyl is at positions 1-5, 7-8, 10-11, 13-14, 16-17, and 19-21. None of the rejected claims in the instant application requires the exact structure of the alleged “innovative” design that was allegedly reduced to practice prior to the provisional application filing date. Hence, even if the instant claims were to be directed to a composition, the evidence pointed out by the declarants is not commensurate in scope with the claims. Note that the “objective evidence of nonobviousness must be commensurate in scope with the claims which the evidence is offered to support.” See MPEP §716.02.
Moreover, applicant’s attention is directed to the fact that unexpected results must be compared to the closet prior art. In the instant case, the closest prior art pertaining to the LNA/OMe PS-modified anti-miRNA oligonucleotide is the design of Lennox’s “2’OMe/LNA PS” having LNA at every third position starting from position 2; 2’-O-methyl at other positions; and all phosphorothioate linkages.
Applicant additionally asserts that the claims are not obvious because “the prior art fails to provide the motivation to use” the claimed inhibitor for treating sepsis with a reasonable expectation of success because the disease in dos Santos is not sepsis but rather ARDS. In response, applicant’s attention is directed to the fact that it was art-recognized knowledge that sepsis leads to ARDS as evidenced by dos Santos’s teaching that “sepsis is the leading cause of acute respiratory distress syndrome (ARDS), the clinical syndrome of acute lung injury (ALI) in humans.” See page 1. Furthermore, dos Santos taught that increased expression level of miR-193b-5p has “clinical relevance” in sepsis-induced ARDS and that miR-196b-5p is a “therapeutic target for sepsis-induced ALI.” See pages 2 and 8. In addition, dos Santos’ mice treated with LPS was an art-recognized animal model for studying sepsis as amply evidenced by the cited references in the instant rejection. In fact, claim 4 expressly requires treatment of ARDS which is “induced by the sepsis.” As such, the examiner fails to understand how applicant’s arguments attacking dos Santos’ ARDS as being irrelevant to and different from sepsis can possibly show the asserted nonobviousness of the claims when claim 4 requires ARDS treatment, wherein ARDS is induced by sepsis.
In view of the foregoing, applicant’s arguments and the §1.132 declaration are not found persuasive to support the alleged nonobviousness of the instantly claimed subject matter over the combined teachings cited in the instant rejection and the level of skills/techniques disclosed therein.
Claims 3, 18-22, 24-28, and 33 are rejected under 35 U.S.C. 103 as being unpatentable over dos Santos et al. (European Respiratory Journal, published on January 6, 2022, 59:2004216, of record), Fourie (US 2006/0111398 A1, of record), Thompson et al. (US 2014/0371299 A1, of record), Mazzone et al. (US 2021/0087554 A1, of record), and Lennox et al. (Molecular Therapy – Nucleic Acids, 2013, 2:e117) as applied to claim 1 above, and further in view of Kopecheck et al. (Advances in Bioengineering, 2015, Chapter 4:79-116, of record), Patel et al. (WO 2018/170336 A1, of record), and Chen et al. (Journal of Controlled Release, 2018, 286:46-54, of record).
The teachings of each of dos Santos, Fourie, Thompson, Mazzone, and Lennox as well as the obviousness of claim 1 are described herein above, which are fully incorporated by reference herein thus will not be repeated.
The cited references above do not teach use of microbubbles, the LNP, and a prodrug recited in the rejected claims.
Kopecheck teaches, “In addition to the synergistic effects of ultrasound and MBs to enhance the permeability of biological barriers such as cell membranes, small blood vessels, and the BBB, as discussed above, MBs can serve as protective drug carriers. Drugs can be pre-incorporated into carriers such as liposomes, micelles, or microspheres, and these structures can then be easily attached to lipid MBs”, which “have extremely high drug loading capacities” and “the encapsulated agents can be released during the ultrasound-triggered MB destruction process.” See page 92.
Kopecheck teaches “drug-containing LNPs are physically attached to the surface of MB”, which “possess a high drug loading capacity of the nanocarriers and the systemic targeting capability and vascular permeabilizing effect of bubbles.” See page 99.
Patel teaches an LNP comprising an ionizable cationic, amino lipid (DLin-MC3-DMA), a phospholipid (e.g., phosphatidylcholine), a structural lipid (e.g., a sterol), and a PEG lipid (e.g., PEG-DMG) at the mol% ratio of 45:15:38.5:1.5 for encapsulating an RNA including miRNA and antisense RNA, wherein the LNP can comprise a second sterol (e.g., stigmasterol) that is different from the first sterol (e.g., cholesterol), wherein the LNP comprising the ionizable amino lipid and two sterols provides “improvements in safety, efficacy, and specificity” thus can be used to deliver a therapeutic RNA to a mammalian organ for treating a disease. See paragraphs 0004, 0013, 0015, 0051, 00242, 00258-00261, 00266, and 00272.
Chen teaches, “Lipid nanoparticles (LNP) that contain ionizable amino-lipids are the most clinically advanced delivery system for nucleic acid therapeutics. LNP systems can give rise to “flu-like” symptoms and hypotension from the activation of toll-like receptors and increases in serum cytokine levels even when they contain a payload (e.g. siRNA) that has been engineered to minimize immunostimulatory potential…Thus, the potential immunostimulatory properties of LNP and other formulations of genetic drugs are a major challenge for clinical advancement of gene therapies in general.” See page 46.
Chen discloses that ionizable amino-lipids include DLin-MC3-DMA and “the incorporation of a dexamethasone prodrug directly into LNP containing various types of nucleic acid cargos can greatly reduce the level of pro-inflammatory cytokines induced by the LNP.” See pages 46-47.
Chen reports, “As little as 0.5 mg/kg of dexamethasone equivalent as a prodrug in LNP results in essentially complete suppression of pro-inflammatory cytokines such as KC-GRO, TNFa, IL-1b and IL-6 in the immunostimulatory LNP-CpG model”.
It would have been obvious to one of ordinary skill in the art before the effective filing date to encapsulate the LNA/2’-O-methyl/PS-modified miR-193b-5p inhibitor that is rendered obvious above in Patel’s LNP and to further include Chen’s dexamethasone prodrug in the LNP. One of ordinary skill in the art would have been motivated to do so with a reasonable expectation of success in order to provide improved in vivo delivery of the miR-193b-5p inhibitor with reduced immune stimulation induced by the LNP encapsulating the miR-193b-5p inhibitor for providing effective sepsis treatment because LNPs comprising ionizable amino lipids such as DLin-MC3-DMA were deemed “the most clinically advanced delivery system for nucleic acid therapeutics” as reported by Chen thus use of such LNPs for sepsis treatment purpose would have been deemed desirable, and because ionizable amino lipid-containing LNPs comprising two different sterols were taught to provide “improvements in safety, efficacy, and specificity” for in vivo delivery of a therapeutic RNA for disease treatment purpose as taught by Patel. Further, it was art-recognized knowledge that LNPs encapsulating therapeutic RNAs have “immunostimulatory properties” that increase cytokine levels and it was demonstrated in the relevant art that LNPs co-encapsulating both the therapeutic RNAs and dexamethasone prodrug reduce pro-inflammatory cytokine levels induced by the LNPs thus an LNP further comprising dexamethasone prodrug in addition to a therapeutic RNA was deemed therapeutically useful as evidenced by Chen. As such, further incorporating Chen’s dexamethasone prodrug in the LNP encapsulating the miR-193b-5p inhibitor would have been a desirable, obvious option for one of ordinary skill in the art to pursue for reducing the LNP-induced pro-inflammatory cytokine activation, especially for the purpose of treating sepsis, which was known as a “systemic inflammatory response syndrome (“SIRS”)” that “can be simply defined as a spectrum of clinical conditions caused by the immune response of a patient to infection”, wherein suppression of inflammatory responses caused by LNP would thus have been deemed even more desirable for treating the “systemic inflammatory response syndrome”. Since the prior art’s ionizable cationic lipid included in the LNP is structurally indistinguishable from that claimed in the instant case, it necessarily follows that the prior art’s ionizable cationic lipid inherently has the property of being positively charged in a formulating buffer at a pH of about 3 to about 5.5 and the property of being neutral in a storage buffer having a pH of about 7 to about 8, absent objective evidence to the contrary.
It would also have been obvious to one of ordinary skill in the art to attach the LNP encapsulating the miR-193b-5p inhibitor to the surface of microbubbles. One of ordinary skill in the art would have been motivated to do so with a reasonable expectation of success in order to further protect the therapeutic miR-193b-5p inhibitor in vivo and also to further enhance intracellular delivery/availability of the therapeutic miR-193b-5p inhibitor, thereby improving the efficacy of sepsis treatment because it was known in the art that microbubbles “can serve as protective drug carriers” and the combination of microbubbles and ultrasound that triggers drug release from microbubbles was known to provide “synergistic effects” that “enhance the permeability of biological barriers such as cell membranes” as evidenced by the teachings of Kopecheck.
In view of the foregoing, claims 3, 18-22, and 24-28 taken as a whole would have been prima facie obvious before the effective filing date.
Claims 8 and 29-30 are rejected under 35 U.S.C. 103 as being unpatentable over dos Santos et al. (European Respiratory Journal, published on January 6, 2022, 59:2004216, of record), Fourie (US 2006/0111398 A1, of record), Thompson et al. (US 2014/0371299 A1, of record), Mazzone et al. (US 2021/0087554 A1, of record), and Lennox et al. (Molecular Therapy – Nucleic Acids, 2013, 2:e117) as applied to claim 1 above, and further in view of Ektesabi et al. (Shock, 2021, 56:133-141, of record) and Noyes et al. (US 2019/0175506 A1, of record)
The teachings of each of dos Santos, Fourie, Thompson, Mazzone, and Lennox as well as the obviousness of claim 1 are described herein above, which are fully incorporated by reference herein thus will not be repeated.
The cited references above do not teach miR-187-3p and a nanovesicle.
Ektesabi reports that “transfection of primary cardiomyocytes with the miR187-3p resulted in decreased expression of ITPKC, LRRC59, and TBL1 in LPS-treated cardiomyocytes” and that “miR-187-induced regulation of immune receptors and inflammatory pathways mitigates the dysregulated inflammatory response of the heart to mediate MSC-dependent cardio-protective effects, which may improve survival in septic mice treated with MSCs.” See pages 137 and 140.
Noyes teaches that a nanovesicle is a cell-derived small vesicle comprising a membrane that comprises lipids, which can be loaded with a payload such as a therapeutic nucleic acid molecule (e.g., “miRNA”, “siRNA”, “modified antisense oligonucleotide”) having regulatory function thus the nanovesicle can be used as a pharmaceutical composition. See paragraphs 0021-0022, 0026, 0052, 0068, 0077, 0083, and 0085.
It would have been obvious to one of ordinary skill in the art before the effective filing date to formulate a cell membrane-derived nanovesicle encapsulating both the LNA/2’-O-methyl/PS-modified miR-193b-5p inhibitor and Ektesabi’s miR-187-3p mimic and administer the nanovesicle to an LPS-induced sepsis animal model and observe extended survival and reduced organ dysfunction. One of ordinary skill in the art would have been motivated to use a nanovesicle for encapsulating the prior art’s agents with a reasonable expectation of success because the cell membrane-derived nanovesicle was an art-recognized nucleic acid drug carrier that can be formulated as a pharmaceutical composition as taught by Noyes. One of ordinary skill in the art would have been motivated to encapsulate both the LNA/2’-O-methyl/PS-modified miR-193b-5p inhibitor and Ektesabi’s miR-187-3p mimic in Noyes’ nanovesicle with a reasonable expectation of success in order to provide a greater improvement in survival and organ dysfunction reduction in the LPS-induced sepsis animal model compared to administration of a single agent because each of miR-193b-5p inhibitor and Ektesabi’s miR-187-3p mimic was shown to reduce inflammation in the LPS-induced sepsis animal model thus, one of ordinary skill in the relevant art would have reasonably expected a greater level of reduced systemic inflammation in the LPS-induced sepsis animal model when both of miR-193b-5p inhibitor and miR-187-3p mimic are administered, wherein such greater level of reduction in systemic inflammation would have been reasonably expected to provide an improved survival and improved organ dysfunction reduction in the LPS-induced sepsis animal model. Since it was art-recognized knowledge that LPS-induced inflammation includes “multiple organ dysfunction syndrome,” “acute respiratory distress syndrome,” and “neuroinflammation” as taught by Fourie, one of ordinary skill in the art would have applied the combination method comprising administering a nanovesicle encapsulating the two anti-inflammatory RNAs for the purpose of treating any of the art-recognized sepsis-related conditions.
In view of the foregoing, claims 8 and 29-30 taken as a whole would have been prima facie obvious before the effective filing date.
Conclusion
No claim is allowed.
Applicant's amendment necessitated the new ground(s) of rejection presented in this Office action. Accordingly, THIS ACTION IS MADE FINAL. See MPEP § 706.07(a). Applicant is reminded of the extension of time policy as set forth in 37 CFR 1.136(a).
A shortened statutory period for reply to this final action is set to expire THREE MONTHS from the mailing date of this action. In the event a first reply is filed within TWO MONTHS of the mailing date of this final action and the advisory action is not mailed until after the end of the THREE-MONTH shortened statutory period, then the shortened statutory period will expire on the date the advisory action is mailed, and any nonprovisional extension fee (37 CFR 1.17(a)) pursuant to 37 CFR 1.136(a) will be calculated from the mailing date of the advisory action. In no event, however, will the statutory period for reply expire later than SIX MONTHS from the mailing date of this final action.
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/DANA H SHIN/Primary Examiner, Art Unit 1635
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