DETAILED ACTION
The present application, filed on or after March 16, 2013, is being examined under the first inventor to file provisions of the AIA .
Claim(s) 1-9 are pending and under consideration.
Priority
Acknowledgement is made that this application claims foreign priority to KR10-2023-0059825 filed 05/09/2023.
Receipt is acknowledged of certified copies of papers required by 37 CFR 1.55.
Information Disclosure Statement
Receipt of the information disclosure statement on 01/13/2026 is/are acknowledged. The signed and initialed PTO-1449 form(s) has/have been mailed with this action.
Drawings
The drawings are objected to because of the following:
Figure 2 graphs are illegible (i.e., X- and Y- axis labels).
Figure 4 graph is illegible (i.e., X- and Y- axis labels).
The legend of Figure 5 associated with the images is illegible.
The conditions in Figure 6C cannot be made out (i.e., the lines are all the same shade).
Figure 8 graphs are illegible (i.e., X- and Y- axis labels).
Figure 9A graph is illegible (i.e., X- and Y- axis labels).
Figure 9B graph is illegible (i.e., X- and Y- axis labels).
Corrected drawing sheets in compliance with 37 CFR 1.121(d) are required in reply to the Office action to avoid abandonment of the application. Any amended replacement drawing sheet should include all of the figures appearing on the immediate prior version of the sheet, even if only one figure is being amended. The figure or figure number of an amended drawing should not be labeled as “amended.” If a drawing figure is to be canceled, the appropriate figure must be removed from the replacement sheet, and where necessary, the remaining figures must be renumbered and appropriate changes made to the brief description of the several views of the drawings for consistency. Additional replacement sheets may be necessary to show the renumbering of the remaining figures. Each drawing sheet submitted after the filing date of an application must be labeled in the top margin as either “Replacement Sheet” or “New Sheet” pursuant to 37 CFR 1.121(d). If the changes are not accepted by the examiner, the applicant will be notified and informed of any required corrective action in the next Office action. The objection to the drawings will not be held in abeyance.
Specification
Minor informalities
The disclosure is objected to because of the following informalities:
(1) As defined on page 8, line 11, the miRNA of the invention is “MACC1-specic miRNA.” The following locations recite, “. . . MACC1 miRNA of the present disclosure”, (p. 6, lines 3, 5, 10, 14, 15, 22; and p.7, lines 2, 6, 10, and 14). It would be remedial to change “MACC1 miRNA” to “MACC1-specific miRNA” for consistency.
(2) There are two periods ending the sentence on page 17 line 6. It would be remedial to remove one.
Appropriate correction is required.
Trademarks/Tradenames
The use of the following term(s) which is a trade name or a mark used in commerce, has been noted in this application.
IVIS (p.6, line 20 and p.25, line 13);
Primojel (p.15, line 15);
Tween (p.17, lines 13, 23; p.18, line 3);
Witespol (p.17, line 25);
Tego (p.18, line 4);
Exo-Fect (p.22, line 13); and m
Zetaview (p. 22, line 16);
The term should be accompanied by the generic terminology; furthermore the term should be capitalized wherever it appears or, where appropriate, include a proper symbol indicating use in commerce such as ™, SM , or ® following the term.
Although the use of trade names and marks used in commerce (i.e., trademarks, service marks, certification marks, and collective marks) are permissible in patent applications, the proprietary nature of the marks should be respected and every effort made to prevent their use in any manner which might adversely affect their validity as commercial marks.
Browser-Executable code
The disclosure is objected to because it contains an embedded hyperlink and/or other form of browser-executable code. Applicant is required to delete the embedded hyperlink and/or other form of browser-executable code; references to websites should be limited to the top-level domain name without any prefix such as http:// or other browser-executable code. See MPEP § 608.01.
Page 11, line 18
Claim Objections
Claim 6 is objected to because of the following informalities: it would be remedial to put “of” in front of SEQ ID NO:3.
Claim 9 is objected to because of the following informalities: it would be remedial to put “a” in front of “pharmaceutical composition” to recite “wherein the composition is a pharmaceutical composition.”
Appropriate correction is required.
Claim Rejections - 35 USC § 112(a) - Written Description
The following is a quotation of the first paragraph of 35 U.S.C. 112(a):
(a) IN GENERAL.—The specification shall contain a written description of the invention, and of the manner and process of making and using it, in such full, clear, concise, and exact terms as to enable any person skilled in the art to which it pertains, or with which it is most nearly connected, to make and use the same, and shall set forth the best mode contemplated by the inventor or joint inventor of carrying out the invention.
The following is a quotation of the first paragraph of pre-AIA 35 U.S.C. 112:
The specification shall contain a written description of the invention, and of the manner and process of making and using it, in such full, clear, concise, and exact terms as to enable any person skilled in the art to which it pertains, or with which it is most nearly connected, to make and use the same, and shall set forth the best mode contemplated by the inventor of carrying out his invention.
Claim(s) 1-5 and 7-9 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.
The fundamental factual inquiry is whether the specification conveys with reasonable clarity to those skilled in the art that, as of the filing date sought, Applicant was in possession of the invention as now claimed. See, e.g., Vas-Cath, Inc., 935 F.2d at 1563-64, 19 USPQ2d at 1117.
Claim(s) 1-9 are drawn to a genus of MACC1 inhibitor for treating colon cancer. The rejected claims thus comprise a genus of MACC1 inhibitors and are defined as belonging to the broad class of MACC1 inhibitors and as having the function of treating colon cancer.
Claim 4 is drawn to subgenera of MACC1 inhibitors, e.g., miRNA, shRNA, siRNA, ribozyme, DNAzyme, PNA, an antibody, and an aptamer. The rejected claim thus comprises subgenera of MACC1 inhibitors and are defined as belonging to the broad class of MACC1 inhibitors and as having the function of treating colon cancer.
Claim 5 is drawn to the species, of the subgenus of miRNA, of the genus of MACC1 inhibitors, i.e., miR143. The rejected claim thus comprises a species of MACC1 inhibitor and is defined as belonging to the broad class of MACC1 inhibitors and having the function of treating colon cancer.
Claim 6 is drawn to a subspecies of the species of miR143, i.e., SEQ ID NO: 3. The rejected claim thus comprises a subspecies of MACC1 inhibitor and is defined as belonging to the broad class of MACC1 inhibitors and having the function of treating colon cancer.
To satisfy the written description requirement, MPEP §2163 states, in part “… a patent
specification must describe the claimed invention in sufficient detail that one skilled in the art can reasonably conclude that the inventor had possession of the claimed invention.” Moreover, the written description requirement for a genus may be satisfied through sufficient description of a representative number of species by “… disclosure of relevant, identifying characteristics, i.e.,
structure or other physical and/or chemical properties, by functional characteristics coupled with
a known or disclosed correlation between functional and structure, or by a combination of such
identifying characteristics, sufficient to show the applicant was in possession of the claimed genus.”
The specification envisions the MACC1 inhibitors as the following:
“In an embodiment of the present disclosure, the inhibitor may be one selected from the group consisting of miRNA, short hairpin RNA (shRNA), small interference RNA (siRNA), ribozyme, DNAzyme, PNA, an antibody, and an aptamer, but is not limited thereto. In an embodiment of the present disclosure, the inhibitor may be miR143, but is not limited thereto. In an embodiment of the present disclosure, the miR143 may comprise SEQ ID NO: 3, but is not limited thereto.” (p. 4 lines 6-13; and p. 9, lines 14-21).
“In the present disclosure, "inhibitor" may include anything that inhibits the expression of a specific substance, especially a protein, or suppresses the activity of its function. Accordingly, the MACC1 inhibitor of the present disclosure may include all substances that inhibit the expression of MACC1 protein or inhibit MACC1 protein activity, or substances that inhibit MACC1 protein expression and activity.”, (p. 8, lines 1-5).
Also see table 2 for SEQ ID NO:3.
The specification does not describe any of the claimed (a) shRNA, (b) siRNA, (c) ribozyme, (d) DNAzyme, (e) PNA, (f) an antibody, or (g) an aptamer in a way that is sufficient to treat colon cancer. The specification does not describe more than one miRNA that is sufficient to treat colon cancer. The specification does not provide description for other MACC1 inhibitors i.e., upstream regulators or downstream feedback loops, or other means of targeting MACC1 through inhibitors such small molecules, natural compounds, biologics (other than an antibody) etc., capable of treating colon cancer.
Even if one accepts that the examples described in the specification meet the claim limitations of the rejected claims in regard to structure and function, the examples are only representative of one subspecies of MACC1 inhibitor (i.e., SEQ ID NO: 3) used for treating colon cancer. These results are not necessarily predictive of all “MACC1 inhibitors” capable of treating colon cancer. Thus, it is impossible for one to extrapolate from the one example of MACC1 inhibitor described herein that the genus of MACC1 inhibitors would necessarily meet the structural/functional characteristics of the rejected claims.
The prior art does not appear to offset the deficiencies of the instant specification in that it does not describe a set of “MACC1 inhibitors” capable of treating colon cancer.
Schope et al (MACC1 revisited - an in depth review of a master of metastasis; Biomark Res, vol 12, issue 146, pages 1-26; published November 23rd, 2024) teaches MACC1 cellular role, inhibitors of MACC1, and the future of cancer therapeutics.
Schope et al teaches in Figure 1 that MACC1 is not only involved in colon cancer but also in a myriad of other cancers such as retinoblastoma, nasopharyngeal, breast, liver, gallbladder, pancreatic, endometrial, cervical, glioblastoma, head & neck, tongue, esophageal, lung, hepatobiliary, renal, gastric, melanoma, ovarian, bladder, and osteosarcoma.
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Schope et al teaches, “Moreover, the expression levels can greatly vary, ranging from almost complete absence to massive overexpression. Contrastingly, the expression of MACC1 is hardly detected in normal tissues and its physiological function remains largely unknown. . . Moreover, the expression of MACC1 and its functions are deeply entangled with pathways of membrane-bound receptors for growth, inflammation and cell death [34, 73] (Fig. 2). Specifically, MACC1 is closely tied with the following signaling pathways:”, (p.5, col 2, para 2; and see figure 2 below).
“Regulation of MACC1 expression and its impact on signaling pathways. The expression and function of MACC1 is regulated by receptor tyrosine kinases such as HGF/c-MET and cytokine receptors including TNFR1, but also through micro-RNAs and the lncRNA MACC1-AS1, functioning via AMPK. Downstream targets of MACC1 include MEK/ERK, PI3K/Akt, STAT1/3, TWIST/VEGF, Wnt/β-catenin and the stemness genes NANOG/Oct4/LGR5.”, (figure 2 legend; p.6).
As for small molecule inhibitors, Schope et al teaches, “. . . Crocin was capable of downregulating DCLK1 leading to the restriction of MACC1 mediated functions [132]. Furthermore, curcumin—found in turmeric, a member of the ginger family—was recognized to reduce MACC1 expression in a concentration-dependent manner in CRC cell lines, resulting in a reduction of proliferation, migration, clonogenicity and wound healing [216]. Cantharidin—a substance of the terpenoid class secreted by many species of blister beetles—and its analogue norcantharidin, known from traditional Chinese medicine, were investigated for their effect on MACC1. Both agents were able to reduce MACC1 gene expression as well as functional outcomes of MACC1 expression such as migration and colony formation [217]. Conclusively, natural compounds possess the capacity to affecting MACC1 expression levels but additional research is needed to further understand the mechanism by which these compounds facilitate their inhibition on MACC1 and to evaluate their feasibility to supplement modern anti-cancer therapies.”, (p.16, col 1, para 2).
As for microRNAs, Schope et al teaches, “In a therapeutic context, miRNA-338-3p mimics were able to suppress cell proliferation, colony formation, migration, and induced apoptosis in CRC cell lines. . . Using a luciferase assay, Lu et al. showed that MACC1 is a direct target of miRNA-338-3p leading to reduced cell proliferation, migration and G1/S cell cycle arrest [186]. . . Additionally, miRNA-940 [188], miRNA-145-5p [189], miRNA-138-5p [190], miRNA-320a [191], miRNA-1236-3p [192], and miRNA-330-5p [193] have been shown to have a regulative impact on MACC1 expression levels in CRC.”, (p.13, col 2, para 4).
See table 1 for MACC1 expression-effecting molecules (p.18-p.19).
In summary, Schope et al teaches various small molecules and microRNAs that regulate MACC1 expression levels in colorectal cancer cells. Schope et al also teaches that downstream targets of MACC1 include MEK/ERK, PI3K/Akt, STAT1/3, TWIST/VEGF, Wnt/β-catenin and the stemness genes NANOG/Oct4/LGR5, and that upstream regulators include receptor tyrosine kinases such as HGF/c-MET and cytokine receptors including TNFR1, and also micro-RNAs and the lncRNA MACC1-AS1, functioning via AMPK. The instant specification fails to describe inhibitors targeting upstream and/or downstream regulators of MACC1.
Thus, with regard to microRNAs:
Venneri and Passantino (MiRNA: what clinicians need to know, European Journal of Internal Medicine, Vol 113, pages 6-9, published May 20, 2023) teach on microRNA: “Approximately 2500 miRNA genes have been discovered in the human genome. An individual miRNA can modulate the expression and function of hundreds of mRNAs; in contrast, each mRNA can be regulated by multiple miRNAs. The interaction between miRNAs and their mRNA targets involves base-pairing of 6–8 nucleotide sequences. As much as 60% of the human transcriptome is considered to be negatively regulated by miRNA activity.”, (page 6, column 1, paragraph 2).
Further, “Functions of miRNAs. MiRNAs are involved in many biological functions, specifically the following.
• Developmental regulation: miRNAs are involved in regulating gene expression during development, including cell differentiation and organogenesis.
• Cellular homeostasis: miRNAs play roles in maintaining cellular homeostasis by regulating various cellular processes, such as proliferation, apoptosis, and stress responses.
• Immune response: miRNAs are involved in regulating immune responses by controlling the expression of genes involved in innate and adaptive immunity.
• Metabolic regulation: miRNAs play roles in the regulation of metabolic processes such as glucose and lipid metabolism.
• Neurological function: miRNAs are abundant in the central nervous system and have crucial roles in neuronal development, including neurogenesis, synapse formation, neural plasticity, and axon guidance.
• Cardiovascular function: miRNAs participate in both heart development and heart function, including cardiac development, angiogenesis, and vascular remodeling.
• Epigenetic regulation: miRNAs regulate epigenetic modifications such as DNA methylation and histone modifications.”, (page 6, column 1 paragraph 5 to column 2 paragraph 1).”
Menon et al (miRNA: A Promising Therapeutic Target in Cancer, Int Jour of Molecular Sciences, vol 23, issue 11502, pages 1-29, published September 29th, 2022) teaches:
“To date, researchers have not developed any cancer therapy that can wipe off cancer cells from the system. With advancements in research, we are now able to elucidate the mechanisms of cancer initiation and progression at the molecular level. This has shed light on the tight link between miRNAs and cancer, and miRNA therapeutics can be the future of cancer therapy. The main hurdle that is faced in miRNA-based therapy is target specificity. As one miRNA can target many genes, there will be off-target effects which need to be addressed. It has been reported that off-target gene silencing can lead to neuro and immunotoxicity and may reduce its therapeutic effects [210]. Intensive studies will be necessary to elucidate the multiple gene targets of miRNAs and to study the off-target effects. A study completed in D. melanogaster emphasizes the off-target issues of RNAi techniques and suggests to use co-inhibition or co-induction techniques [211]. Targeting a gene with a single miRNA would not be sufficient to bring out the desired effect. Co-inhibition or co-induction using a combination of miRNAs could be a possible solution for this, and it may be helpful to provide a cumulative effect. In general, a deep understanding of the interaction between the miRNA and the target genome and the signaling pathways that are modulated would be beneficial in experimental designs.”, (p.20, para 1 of section 10: Future Prospects and Challenges).
Zhang et al (The risk of miRNA Therapeutics: In a Drug Target Perspective, Drug Design, Development and Therapy, volume 15, pages 721-733, published February 22nd, 2021) teaches: “As expected, Figure 2A showed a flexible complementary ratio of miRNA with target sequence (within the range 20–90%) and none with complete complementation, but all siRNAs had 100% complementary ratio. The less restricted complementary mode may lead to low specificity of target sequence.”, (p.722, col 2, para 2). Moreover, “Beyond our expectation, the targets of the miRNA drug ranged from 30 to 250 in number and almost all miRNA drug* were over 500 and even 1000, but the si0052NA drug generally targets 1–3 genes (Figure 2B). The majority of miRNA targets tens and hundreds of genes, and we named it “too many targets for miRNA effect” (TMTME).”, (p.722, col 2, para 2).
Further, Zhang et al teaches, “TMTME (“too many targets for miRNA effect”) is a typical and inevitable property of miRNA molecules, which is caused by incomplete complementation with the target sequence. TMTME leads to that miRNA could bind to various sequences suitable for the interaction (including protein-coding genes, lncRNA, circRNA, etc), which is different from all approved drugs (including siRNA drugs) with only a few targets… Therefore, both introduction and removal of miRNA in humans can lead to changes of a wide series of pathways and some of them are unknown, even unpredictable, probably triggering disorders of physiological function or the occurrence of additional disease.”, (Page 729, column 1, paragraph 2 to column 2, paragraph 1).
Zhang et al teaches, “Moreover, due to instability of unprotected miRNAs, delivering miRNAs required chemical modifications to avoid rapid degradation in serum, which may impair specificity of miRNAs and lead to off-target effects… Another challenge is that exogenous artificial miRNAs will trigger competition and saturation effect, a competition among exogenous and the endogenous miRNAs for the intracellular machinery, and thus affecting unexpected gene expression and leading to untoward side effects.”, (page 730, column 1, paragraph 1).
Thus, the instant specification fails to describe and/or disclose:
A set of MACC1 inhibitors representative of the direct and indirect pathways
MACC1 is involved in (e.g., inhibitors that target receptor tyrosine kinases such as HGF/c-MET and cytokine receptors including TNFR1, and lncRNA MACC1-AS1, functioning via AMPK; and/or downstream target modulation for a potential feedback loop mechanism of MACC1 via MEK/ERK, PI3K/Akt, STAT1/3, TWIST/VEGF, Wnt/β-catenin and the stemness genes NANOG/Oct4/LGR5) for the treatment of colon cancer;
A set of microRNAs representative of the subgenus for treating colon cancer. As
taught by Venneri and Passantino, there are about 2500 microRNAs with functions ranging from regulation of cell differentiation and organogenesis to proliferation and apoptosis to glucose and lipid metabolism to DNA methylation and histone modification, all outside the function prescribed by the claimed invention of treatment for colon cancer. Claiming “microRNA” is not predictive of all the microRNA(s) ability to treat colon cancer; and
Therefore, the art does not appear to offset the deficiencies of the specification. Merely describing a “MACC1 inhibitors” capable of treating colon cancer without sufficient detail relating to the genus of MACC1 inhibitors in the treatment of colon cancer does not allow the skilled artesian to reasonably conclude that the Applicants were in possession of the claimed invention in claim(s) 1-5 and 7-9.
Claim Rejections - 35 USC § 112 – Lack of Enablement
The following is a quotation of the first paragraph of 35 U.S.C. 112(a):
(a) IN GENERAL.—The specification shall contain a written description of the invention, and of the manner and process of making and using it, in such full, clear, concise, and exact terms as to enable any person skilled in the art to which it pertains, or with which it is most nearly connected, to make and use the same, and shall set forth the best mode contemplated by the inventor or joint inventor of carrying out the invention.
The following is a quotation of the first paragraph of pre-AIA 35 U.S.C. 112:
The specification shall contain a written description of the invention, and of the manner and process of making and using it, in such full, clear, concise, and exact terms as to enable any person skilled in the art to which it pertains, or with which it is most nearly connected, to make and use the same, and shall set forth the best mode contemplated by the inventor of carrying out his invention.
Claim(s) 1-9 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 enablement requirement. The claim(s) contains subject matter which was not described in the specification in such a way as to enable one skilled in the art to which it pertains, or with which it is most nearly connected, to make and/or use the invention.
Enablement is considered in view of the Wands factors (MPEP 2164.01(A)). These include: the breadth of the claims, the nature of the invention, the state of the prior art, the level of one of ordinary skill, the level of predictability in the art, the amount of direction provided by the inventor, the existence of working examples, and the quantity of experimentation needed to make or use the invention. All of the Wands factors have been considered with regard to the instant claims, with the most relevant factors discussed below.
Nature of the invention:
Claim 1 is drawn to a method of treating colon cancer, the method comprising: administering extracellular vesicles loaded with an MACC1 inhibitor or a composition comprising the extracellular vesicles as an active ingredient to a subject in need thereof. The nature of the invention is complex in that “inhibitor” encompasses a broad genus of chemical(s) used to reduce expression and/or activity of MACC1, which can be direct or indirect, via a small molecule, an antibody, a nucleic acid, etc. All of which must be capable of treating colon cancer in a subject in need thereof.
Claim 2 and 3 are drawn to wherein the extracellular vesicles comprise a TM4SF5-targeting peptide (of claim 2) and wherein the peptide comprises the amino acid sequence of SEQ ID NO:1 (of claim 3). The nature of the invention is complex in that extracellular vesicles comprise a targeting peptide (i.e., comprising SEQ ID NO:1) and must be capable of encompassing a broad genus “MACC1 inhibitor” which include chemical(s) used to reduce expression and/or activity of MACC1, which can be direct or indirect, via a small molecule, an antibody, a nucleic acid, etc. All of which must be capable of treating colon cancer in a subject in need thereof.
Claim 4 is drawn to wherein the inhibitor is one selected from the group consisting of miRNA, shRNA, siRNA, ribozyme, DNAzyme, PNA, an antibody and an aptamer. The nature of the invention is complex in that it encompasses the broad subgenera of the inhibitors listed above and those inhibitors must be capable of treating colon cancer in a subject in need thereof.
Claim 5 and claim 6 are drawn to wherein the inhibitor is mir143 (of claim 5) and wherein the mir143 comprises the nucleotide sequence SEQ ID NO: 3 (of claim 6). The nature of the invention is complex in that it encompasses a species and subspecies of microRNA that must be capable of treating colon cancer in a subject in need thereof.
Claim(s) 7-9 are drawn to wherein the extracellular vesicles are derived from adipose-derived stem cells (of claim 7), wherein the composition is administered intravenously (of claim 8), and wherein the composition is a pharmaceutical composition (of claim 9). The nature of the invention is complex in that extracellular vesicles derived from adipose-derived stem cells, the administration intravenously, and a pharmaceutical composition, must be capable of encompassing a broad genus “MACC1 inhibitor” which include chemical(s) used to reduce expression and/or activity of MACC1, which can be direct or indirect, via a small molecule, an antibody, a nucleic acid, etc. All of which must be capable of treating colon cancer in a subject in need thereof.
Breadth of the claims:
The broadest reasonable interpretation of claim 1 is that it encompasses a method of treating colon cancer, the method comprising: administering extracellular vesicles loaded with any/all MACC1 inhibitor(s) or a composition comprising the extracellular vesicles as an active ingredient to a subject in need thereof.
The complex nature of the subject matter of this invention is greatly exacerbated by the breadth of the claims.
Guidance of the specification and existence of working examples:
Looking to the specification for treating colon cancer, MACC1 inhibitors, and working examples.
Treatment of colon cancer:
“In the present disclosure, "prevention" refers to all actions that inhibit or delay the onset of a target disease; "treatment" refers to all actions that improve or beneficially change a target disease and metabolic abnormalities thereof by the administration of the pharmaceutical composition according to the present disclosure; and "alleviation" refers to all actions that reduce target discase-related parameters, for example, the degree of symptoms, by the administration of the pharmaceutical composition according to the present disclosure.”, (p.20, lines 10-16).
MACC1 inhibitors:
“In an embodiment of the present disclosure, the inhibitor may be one selected from the group consisting of miRNA, short hairpin RNA (shRNA), small interference RNA (siRNA), ribozyme, DNAzyme, PNA, an antibody, and an aptamer, but is not limited thereto. In an embodiment of the present disclosure, the inhibitor may be miR143, but is not limited thereto. In an embodiment of the present disclosure, the miR143 may comprise SEQ ID NO: 3, but is not limited thereto.” (p. 4 lines 6-13; and p. 9, lines 14-21).
“In the present disclosure, "inhibitor" may include anything that inhibits the expression of a specific substance, especially a protein, or suppresses the activity of its function. Accordingly, the MACC1 inhibitor of the present disclosure may include all substances that inhibit the expression of MACC1 protein or inhibit MACC1 protein activity, or substances that inhibit MACC1 protein expression and activity.”, (p. 8, lines 1-5).
Also see Table 2.
Working examples:
Example 1: making and confirming that the extracellular vesicles contain miR143.
Example 2: confirming MACC1 expression after treating colon cancer cell lines with control and mir143 vesicles (HCT116 and HT29 cells used). Bax and MCL-1 were also measured.
Example 3: protein level analysis of the experiment of example 2, i.e., western blot.
Example 4: confirming inhibition of intracellular movement based on the experiment in example 2.
Example 5: confirming whether or not the extracellular vesicles could migrate to colon specific tumor tissues with the colon-cancer targeting peptide in a mouse model of colon cancer.
Example 6: confirmation of reduced tumor sizes in the mouses with the mir143 extracellular vesicles of Example 5.
Example 7: confirmation of inhibition of MACC1 expression, Bax and MCL-1 from the experiment performed in Example 5.
Example 8: protein level analyses of the experiment performed in Example 5.
Example 9: immunohistochemistry of BCL-xL and Bax in the tumors from Example 5.
Predictability and state of the art:
Schope et al (supra) teaches MACC1 cellular role, inhibitors of MACC1, and the future of cancer therapeutics.
Schope et al teaches in Figure 1 that MACC1 is not only involved in colon cancer but also in a myriad of other cancers such as retinoblastoma, nasopharyngeal, breast, liver, gallbladder, pancreatic, endometrial, cervical, glioblastoma, head & neck, tongue, esophageal, lung, hepatobiliary, renal, gastric, melanoma, ovarian, bladder, and osteosarcoma.
Schope et al teaches, “Moreover, the expression levels can greatly vary, ranging from almost complete absence to massive overexpression. Contrastingly, the expression of MACC1 is hardly detected in normal tissues and its physiological function remains largely unknown. . . Moreover, the expression of MACC1 and its functions are deeply entangled with pathways of membrane-bound receptors for growth, inflammation and cell death [34, 73] (Fig. 2). Specifically, MACC1 is closely tied with the following signaling pathways:”, (p.5, col 2, para 2; and see figure 2 in the above rejection).
“In a therapeutic context, miRNA-338-3p mimics were able to suppress cell proliferation, colony formation, migration, and induced apoptosis in CRC cell lines. . . Using a luciferase assay, Lu et al. showed that MACC1 is a direct target of miRNA-338-3p leading to reduced cell proliferation, migration and G1/S cell cycle arrest [186]. . . Additionally, miRNA-940 [188], miRNA-145-5p [189], miRNA-138-5p [190], miRNA-320a [191], miRNA-1236-3p [192], and miRNA-330-5p [193] have been shown to have a regulative impact on MACC1 expression levels in CRC. Moreover, it was demonstrated that targeting MACC1 via miR-940 enhanced the antitumor effect of anlotinib—a small-molecule multi-target tyrosine kinase inhibitor—effectively reducing proliferation and metastasis. Of note, using CRC cells and CRC xenograft mouse models, a strongly elevated effect on the reduction of tumor size and metastasis formation was seen in a combinatorial approach with miR-940 and anlotinib compared to single treatments.”, (p.13, col 2, para 4).
See table 1 for MACC1 expression-effecting molecules (p.18-p.19).
Menon et al (supra) teaches the challenges of microRNA therapy in cancer treatment.
Menon et al teaches, “Tumor-suppressing miRNAs are always downregulated in cancer, which leads to the overexpression of their target oncogenes. Ectopic expression of tumor-suppressing miRNAs can be used to replace the lost miRNA level, aiming to inhibit cellular pathways that support oncogenesis. miRNA mimics are an effective tool that can be used to restore the function of tumor-suppressing miRNAs. miRNA mimics are small, chemically modified (2′-O’methoxy) double-stranded RNA molecules that mimic the endogenous mature miRNA molecules [84].”, (p.10, section 5.2).
Menon et al further teaches, “To date, there are only 10 miRNAs that have progressed to clinical trials, and none of them has been entered into the clinicaltrials.gov database for phase III trials.”, (p.18, para 1).
Lastly, Menon et al teaches, “To date, researchers have not developed any cancer therapy that can wipe off cancer cells from the system. With advancements in research, we are now able to elucidate the mechanisms of cancer initiation and progression at the molecular level. This has shed light on the tight link between miRNAs and cancer, and miRNA therapeutics can be the future of cancer therapy. The main hurdle that is faced in miRNA-based therapy is target specificity. As one miRNA can target many genes, there will be off-target effects which need to be addressed. It has been reported that off-target gene silencing can lead to neuro and immunotoxicity and may reduce its therapeutic effects [210]. Intensive studies will be necessary to elucidate the multiple gene targets of miRNAs and to study the off-target effects. A study completed in D. melanogaster emphasizes the off-target issues of RNAi techniques and suggests to use co-inhibition or co-induction techniques [211]. Targeting a gene with a single miRNA would not be sufficient to bring out the desired effect. Co-inhibition or co-induction using a combination of miRNAs could be a possible solution for this, and it may be helpful to provide a cumulative effect. In general, a deep understanding of the interaction between the miRNA and the target genome and the signaling pathways that are modulated would be beneficial in experimental designs.”, (p.20, para 1 of section 10: Future Prospects and Challenges).
Zhang et al (supra) provides guidance on miRNA risks in therapeutics: “Only 10 obtainable miRNA drugs have been in clinical trials with none undergoing phase III…”, (abstract). Zhang et al goes on to disclose, “Inherently, miRNA is endogenously produced and siRNA is exogenously designed. Designers can exactly endow siRNA giving them the purpose of gene silencing, while endogenous miRNA seemed more complicated because nobody assigned them specific tasks.”, (see parge 722, column 2, paragraph 2).
“As expected, Figure 2A showed a flexible complementary ratio of miRNA with target sequence (within the range 20–90%) and none with complete complementation…”, (see page 722, column 2, paragraph 2; and Figure 2A).
“…All ten miRNA drugs had tens and hundreds of unapproved targets…”, (see Figure 3A, and page 727, column 1, paragraph 1).
“TMTME (“too many targets for miRNA effect”) is a typical and inevitable property of miRNA molecules, which is caused by incomplete complementation with the target sequence. TMTME leads to that miRNA could bind to various sequences suitable for the interaction (including protein-coding genes, lncRNA, circRNA, etc), which is different from all approved drugs (including siRNA drugs) with only a few targets… Therefore, both introduction and removal of miRNA in humans can lead to changes of a wide series of pathways and some of them are unknown, even unpredictable, probably triggering disorders of physiological function or the occurrence of additional disease.”, (Page 729, column 1, paragraph 2 to column 2, paragraph 1). Lastly, “Moreover, due to instability of unprotected miRNAs, delivering miRNAs required chemical modifications to avoid rapid degradation in serum, which may impair specificity of miRNAs and lead to off-target effects… Another challenge is that exogenous artificial miRNAs will trigger competition and saturation effect, a competition among exogenous and the endogenous miRNAs for the intracellular machinery, and thus affecting unexpected gene expression and leading to untoward side effects.”, (page 730, column 1, paragraph 1).
Thus, one of skill in the art would appreciate the unpredictability of any/all inhibitors of MACC1, especially any/all microRNAs, for the treatment of colon cancer. The art teaches that MACC1 is involved in multiple signaling pathways and can thus be regulated upstream or downstream by inhibitors working in those paths as well. The art teaches that MACC1 is not only involved in colon cancer but is involved in most cancers as a biomarker of metastasis. The art teaches that the basic cellular role of MACC1 in noncancerous tissue is unknown. The art teaches numerous microRNA that are involved in the regulation of MACC1. The art teaches that both the introduction and/or removal of miRNA in humans can lead to changes of a wide series of pathways (some unknown, some unpredictable). The art teaches that miRNA does not bind with 100% complementarity and has tens to hundreds of targets. Lastly, the art teaches targeting a gene with a single miRNA would not be sufficient to bring out the desired effect.
Amount of experimentation necessary:
The quantity needed to carry out the scope of the invention is large. One would be required to screen direct and indirect inhibitors that broadly cover the various functions of the broad genus of MACC1 inhibitor for the ability to therapeutically treat colon cancer. This broad genus includes and is not limited to siRNA, aptamers, antibodies, small molecules, and miRNAs that directly and indirectly influence MACC1 expression. For instance, success with one microRNA would not guarantee success with any other microRNA, or success with one small molecule does not guarantee success with an antibody. This type of experimentation is not routine in the art and would require a large amount of inventive effort. Further considering that any positive results (e.g., successful treatment of colon cancer) would amount to a significant advancement in the state of the art, additional experimentation required is considered undue.
In view of the breadth of the claims and the lack of guidance provided by the specification as well as the unpredictability of the art, the skilled artisan would have required an undue amount of experimentation to make and/or use the claimed invention. Therefore, claims 1-9 are not considered to be enabled by the instant disclosure.
Conclusion
No claims allowed.
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/L.M.T./Examiner, Art Unit 1637
/Jennifer Dunston/Supervisory Patent Examiner, Art Unit 1637
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