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 .
This Office action is in response to the communication filed 12-16-25.
Claims 1, 4, 5, 8, 11-19, 53-56 are pending in the instant application.
Election/Restrictions
Applicant's election with traverse of Group I, in the reply filed on 12-16-25 is acknowledged.
In light of Applicant’s arguments and upon further consideration, the restriction requirement filed 12-16-25 is hereby withdrawn. Claims 1, 4, 5, 8, 11-19, 53-56 have been rejoined and examined on their merits as set forth below.
Claim Rejections - 35 USC § 112
The following is a quotation of the first paragraph of 35 U.S.C. 112(a):
(a) IN GENERAL.—The specification shall contain a written description of the invention, and of the manner and process of making and using it, in such full, clear, concise, and exact terms as to enable any person skilled in the art to which it pertains, or with which it is most nearly connected, to make and use the same, and shall set forth the best mode contemplated by the inventor or joint inventor of carrying out the invention.
The following is a quotation of the first paragraph of pre-AIA 35 U.S.C. 112:
The specification shall contain a written description of the invention, and of the manner and process of making and using it, in such full, clear, concise, and exact terms as to enable any person skilled in the art to which it pertains, or with which it is most nearly connected, to make and use the same, and shall set forth the best mode contemplated by the inventor of carrying out his invention.
Claims 1, 4, 5, 8, 11-19, 53-56 are rejected under 35 U.S.C. 112, first paragraph, because the specification, while being enabling for SB216763-treated osteoblasts infected with lentiviral vectors containing CMV promoter-driven SMAD1 cDNA or osteoblasts infected with lentiviral vectors containing β-catenin siRNA, specific gene deletion of GSK3β in osteoblasts, Flkl-eGFP osteoblasts, GFP+CD31+ aortic cells of Osx-GfptgMgp - mice treated with or without SB216763, the study of total aortic calcium of CollalcreERT2GSK3p/loxlxMgp - mice with or without injection of tamoxifen, does not reasonably enable methods for treating chronic kidney disease, diabetes mellitus, or fibrodysplasia ossificans progressiva in a subject, or of treating or preventing vascular calcification, inducing or increasing osteoblastic-endothelial transdifferentiation, or inhibiting or decreasing osteogenesis in a subject in need thereof comprising the administration of the large genus of modulators claimed.
The specification does not enable any person skilled in the art to which it pertains, or with which it is most nearly connected, to make and/or use the invention commensurate in scope with these claims.
The following factors have been considered in determining that the specification does not enable the skilled artisan to make and/or use the invention over the broad scope claimed.
The breadth of the claims:
The claims are drawn to methods of treating chronic kidney disease, diabetes mellitus, or fibrodysplasia ossificans progressiva in a subject or of treating or preventing vascular calcification, inducing or increasing osteoblastic-endothelial transdifferentiation, or inhibiting or decreasing osteogenesis in a subject in need thereof, comprising administering to the subject any agent that inhibits the activity of or decreases the levels of glycogen synthase kinase 3 (GSK3), which GSK3 is optionally GSK3β, any agent that inhibits the activity of or decreases the levels of mothers against decapentaplegic homolog 1 (SMAD1), or any agent that activates or increases the levels of β-catenin, which agent optionally comprises any small molecule, any polypeptide, or any inhibitory polynucleotide optionally comprising any siRNA, shRNA, or antisense RNA molecule, or which agent is a polynucleotide encoding β-catenin.
Teachings in the art and in the specification.
Teachings in the art:
Roberts et al (Nature Rev., Drug Discovery, Vol. 19, pages 673-694 (2020)) teaches on page 673 that “achieving efficient oligonucleotide delivery, particularly to extrahepatic tissues, remains a major translational limitation.”
Kobelt et al (Cancer Gene Therapy in Gene Therapy of Cancer: Methods and Protocols, Methods in Molecular Biology, Vol. 2521, pages 1-15 (Springer Nature 2022)) teach that limitations to cancer gene therapy relate to limitations in gene transfer efficiency (see esp. pages 3-4).
In addition, Osborn et al (Nucleic Acid Therapeutics, Vol. 28, No. 3, pages 128-136 (2018)) state the following about challenges to siRNA delivery on page 128:
…The primary challenge facing the clinical development of small interfering RNAs (siRNA) has been overcoming barriers that impede in vivo delivery. siRNAs are large, polyanionic macromolecules with intrinsically poor pharmacological properties. Unmodified siRNAs have a half-life of less than 5 min in circulation, and they do not permeate intact cellular membranes…
Damase et al (Frontiers in Bioengineering and Biotechnology, Vol. 9, Article 628137, pages 1-24 (2021)) on page 13 also address the challenges of using RNA-based drugs:
Targeted delivery is a major hurdle for effective RNA therapeutics, a hurdle that must be overcome to broaden the application of clinical translation of this type of therapeutic. …There is a need for novel delivery vehicles that will deliver the RNA drug to the site of therapeutic action facilitating the entry of the RNA drug into the cytoplasm where it may exert its effect…
Bost et al (ACS Nano, Vol. 15, pages 13993-14021 (2021)) on page 13993 also address the current challenges of oligonucleotide therapeutics:
…Historically, the largest hindrance to the widespread usage of ON therapeutics has been their inability to effectively internalize into cells and escape from endosomes to reach their molecular targets in the cytosol or nucleus…
[Emphases added][Citations omitted].
Teachings in the specification:
The specification teaches the following:
Figure 1 has eight parts, A-H, and shows high throughput screening identified SB216763 as an inducer of osteoblastic-endothelial transdifferentiation. Part A shows a strategic drawing osteoblastic-endothelial transdifferentiation affecting vascular calcification. Part B shows a high throughput screening that identified SB216763 as an inducer of eGFP expression in Flkl-eGFP osteoblasts as indicated by the black arrow. Part C shows morphology of Flkl-eGFP osteoblasts after treatment with SB216763 (n=9). Scale bar, 50 pm. Part D shows FACS analysis of SB216763-treated osteoblasts using anti-CD31 and anti- VE-cadherin (VE-cad) antibodies (n=8). Part E shows expression of the osteogenic markers Cbfal, osterix (OSX), and osteocalcin (OC) and the endothelial markers CD31, VE-cadherin, CD34, eNOS in osteoblasts treated with different does of SB216763 (n=8). Part F shows time-course expression of the osteogenic markers Cbfal, osterix (OSX), and osteocalcin (OC) and the endothelial markers CD34, VE-cadherin, CD31, and eNOS in SB216763- treated osteoblasts (n=8). Part G shows heatmap and GO analysis of a cohort of genes with decreased expression in SB216763-treated osteoblasts. 827 genes decreased in expression in SB21673-treated osteoblasts were overlapped with 1357 genes identified as low expression when compared osteoblast vs EC, and identified a 307-gene cohort with decreased expression in both analyses. Part H shows heatmap and GO analysis of a cohort of genes with increased expression in SB216763-treated osteoblasts. 519 genes with increased expression after SB21673 treatment were overlapped with 1801 genes with high expression when compared EC vs osteoblast, and identified another 235 genes that showed increased expression in both analyses.
Figure 2 has eleven parts, A-K, and shows SB216763 treatment causes osteoblasts to lose osteogenic capacity but gain endothelial function. Part A shows a schematic of experimental procedure for ectopic bone formation in vivo. HA/TCP, hydroxyapatite / tricalcium phosphate. Part B shows MicroCT images of ectopic bone formation after cell transplantation (n=6). Scale bar, 5 mm. PBS, phosphate buffered saline. Part C shows relative volume of bone formation (n=6). Part D shows implants of ectopic bone collected from the mice after cell transplantation (n=6). Part E shows H&E staining of implants (n=6). Black arrows indicate osteocytes. Scale bar, 50 pm. Part F shows a schematic of experimental procedure for cell transplantation using the hindlimb ischemia model. Part G shows laser4 doppler perfusion images after cell transplantation (n=8). Top, measurement camera. Bottom, documentation camera. PBS, phosphate buffered saline. HAEC, human aortic endothelial cells. Scare bar, 10 mm. Part H shows percentage of blood flow perfusion after cell transplantation normalized by perfusion of normal limb (n=6). Part I shows immunostaining of new vessels after cell transplantation using anti-CD31 antibodies (n=10). Scale bar, 50 pm. Part J shows eGFP positive cells (green) were observed in the endothelium of new vessels, which stained with anti-vWF antibodies (red). Part K shows analysis of vascular density in ischemic sites after cell transplantation (n=10).
Figure 3 has ten parts, A-J, and shows increased β-catenin decreases SMAD1 and
together are responsible for SB216763 to induce osteoblastic-endothelial transdifferentiation. Part A shows a schematic of decreased SMAD1 combined with increased β-catenin caused by GSK3 inhibition converting osteoblasts into endothelial-like cells. Part B shows immunoblotting of SMAD1, phosphorylation (p) SMAD1 and β-catenin in osteoblasts treated with different doses of SB216763 (n=6). Part C shows immunoblotting of SMAD1 and in osteoblasts transfected with β-catenin siRNA in accompany with 10pMSB216763 with (n=6). SCR, scrambled siRNA. Part D shows DNA-binding sites of β-catenin in promoter region of Smad1 gene. Parts E and F show a ChIP assay of DNA-binding of 0-catenin and histone modification in promoter of Smad1 gene. Part G shows microCT images of ectopic bone formation and relative volume of bone formation in the implants after cell transplantation. Osteoblasts were used as controls. SB216763, SB216763-treated osteoblasts. SB216763 / CMV-SMAD1, SB216763-treated osteoblasts infected with lentiviral vectors containing CMV promoter-driven SMAD1 cDNA. SB216763 / 0-catenin si, SB216763- treated osteoblasts infected with lentiviral vectors containing β-catenin siRNA (si) (n=6). Scale bar, 5mm. Part F shows that H&E staining of the sections of implants (n=6). Black arrows indicate osteocytes. Scale bar, 50 pm. Part I shows laser doppler perfusion images after cell transplantation (n=8). Osteoblasts were used as controls. HAEC, human aortic endothelial cells. SB216763, SB216763-treated osteoblasts. SCR, scrambled siRNA. SB216763 / 0-catenin si, SB216763-treated osteoblasts infected with lentiviral vectors containing β-catenin siRNA (si). SB216763 / CMV-SMAD1, SB216763-treated osteoblasts infected with lentiviral vectors containing CMV promoter-driven SMAD1 cDNA. (n=6). Scare bar, 10 mm. Part G shows the percentage of blood flow perfusion after cell transplantation normalized by perfusion of normal limb (n=6).
Figure 4 has two parts, A-B and shows modulation of β-catenin and SMAD 1 alter transcriptional landscapes of osteoblasts. Part A shows a heatmap of the cohorts of genes with decreased SMAD1 DNA-binding and GO analysis of the genes with decreased SMAD1 DNA-binding and decreased expression in SB21673-treated osteoblasts. Part B shows a heatmap of the cohorts of genes with increased p-catenin DNA-binding (bottom) in SB21673-treated osteoblasts and GO analysis of the genes with increased β-catenin DNA- binding and increased expression in SB21673-treated osteoblasts.
Figure 5 has eleven parts, A-K, and shows that SB216763 reverses osteogenesis to
endothelial differentiation to ameliorate vascular calcification in Mgp-/- mice. Part A shows a schematic of experimental procedure to determine the effect SB216763 treatment in early calcification. Part B shows Von Kossa staining of aortic tissues (n=7). Scale bar, 50 pm. Part C shows total aortic calcium after SB216763 treatment (n=8). Part D shows FACS analysis of CD31+ aortic cells using anti-osterix (OSX) and anti-VE-cadherin antibodies (n=6). Part E shows that immunoblotting of aortic tissues of Mgp - mice after SB216763 treatment (n=6). Part F shows schematic of experimental procedure to examine the effect of SB216763 treatment in late calcification. Parts G and H shows microCT images of aortic calcification in Mgp- mice after SB216763 treatment (n=6). Scale bar, 5 mm. Part I shows total aortic calcium and calcification score in Mgp- mice after SB216763 treatment (n=8). Part J shows H&E staining of Mgp - aortic tissues after SB216763 treatment (n=6). Part K shows FACS analysis ofMgp- CD31+ aortic cells using anti-osterix (OSX) and anti-VE-cadherin antibodies (n=6).
Figure 6 has seven parts, A-G, and shows that osteoblastic lineage tracing reveals the shift of osteoblast-like cells to endothelial differentiation in calcified aortic tissue. Part A shows a schematic of experimental procedure to characterize the shift of osteoblast-like cells toward endothelial differentiation. Part B shows FACS analysis of GFP+CD31+ aortic cells of Osx-GfptgMgp - mice treated with or without SB216763 (left). The cells are confirmed to express GFP after isolation (right) (n=5). Scale bar, 50 pm. Part C shows microCT images of ectopic bone formation and analysis of relative volume of bone formation after cell transplantation (n=6). Scale bar, 5 mm. Part D shows H&E staining of implants (n=6). Black arrows indicate osteocytes. Scale bar, 50 pm. Part E shows a laser doppler perfusion images after cell transplantation (n=6). Top, measurement camera. Bottom, documentation camera. Scare bar, 10 mm. Part F shows percentage of blood flow perfusion after cell transplantation normalized by perfusion of normal limb (n=6). Part G shows analysis of vascular density in ischemic sites after cell transplantation (n=10).
Figure 7 has nine parts, A-I, and shows specific gene deletion of GSK3β in osteoblasts reverses osteogenic-endothelial transdifferentiation to ameliorate vascular calcification in Mgp-/- mice. Part A shows immunoblotting of osteoblasts transfected with GSK3α or GSK3β siRNA (n=6). B-actin was used as loading control. Part B shows real-time PCR showing the expression of Cbfal, osterix (OSX), VE-cadherin (VE-cad) and CD31 in osteoblasts transfected with GSK3a or GSK33 siRNA (n=6). Part C shows a schematic of experimental procedure. Part D shows expression of GSK3 in aortic tissues (n=8). Part E shows a total aortic calcium of CollalcreERT2GSK3p/loxlxMgp - mice (n=8) with or without injection of tamoxifen. ff floxflox.Part F and G shows microCT images and aortic calcification score of CollacreERT2GSK3PflOxflOxMgp- mice with or without injection of tamoxifen (n=6). Scale bar, 5 mm. Part H shows H&E staining of CollalcreERT2GSK3p3floxfiMgp - aortic tissues with or without injection of tamoxifen (n=8). Scale bar, 5 mm. Part I shows CD31+ aortic cells isolated from CollalcreERT2GSK3pfloxfoxMgp- aortic tissues with or without injection of tamoxifen and analyzed by FACS analysis using anti-osterix (OSX) and anti-VE-cadherin antibodies (n=6).
Figure 8 has three parts, A-C, and shows that SB216763 treatment has no effect on bone formation. Part A shows that micro-CT imaging of bone tissues of mice treated with or without SB216763 (n=6). Scale bar, 1 mm. Part B shows relative bone volume and connectivity density of bone tissues of mice treated with or without SB216763 (n=6). Part C shows immunostaining of bone tissues by using anti-CD31 or anti-osterix antibodies (n=6). Scale bar, 50pm.
Figure 9 has six parts, A- F, and shows that SB216763 induces endothelial differentiation in human osteoblast but does not activate other lineages. Part A shows high throughput screening identified SB216763 as an inducer of eGFP expression in Flkl-eGFP osteoblasts as indicated by the black arrow. Part B shows a schematic drawing of GSK3 inhibition inducing osteoblastic-endothelial transdifferentiation. Part C shows expression of the osteogenic markers Cbfal, osterix (OSX), and osteocalcin (OC) and the endothelial markers CD31, VE-cadherin, CD34, and eNOS in human osteoblasts treated with different doses of SB216763 (n=8). Part D shows time-course expression of the osteogenic markers Cbfal, osterix (OSX), and osteocalcin (OC) and the endothelial markers CD34, VE-cadherin, CD31, and eNOS in SB216763-treated human osteoblasts (n=8). Part E shows expression of markers for different lineages examined by real-time PCR. The lineages included mesenchymal and stem cell, smooth muscle cell, fibroblast, pericyte, adipocyte, pulmonary, hepatic, neuronal, cardiac, hematopoietic and renal lineages (n=6). a-SMA, alpha smooth muscle actin. MYH11, myosin heavy chain 11. FSP, fibroblast-specific protein. NG2, neuron-glial antigen 2. Spb, surfactant protein b. CCSP, club-cell secretory protein. Aqp5, aquaporin 5. Tnc, troponin c. ACTC1, actin alpha cardiac muscle 1. MYH7, myosin heavy chain 7. AP2, adipocyte protein 2. C/EBP, CCAAT/ enhancer binding protein. Part F shows expression of representative genes in glucose metabolism (n=6). CK2, casein kinase 2.GLUT4, glucose transporter 4. IRS1 and 2, insulin receptor substrates 1 and 2.
Figure 10 has three parts, A-C, and shows SB216763 treatment causes osteoblasts to lose the osteogenic capacity but gain endothelial function. Part A shows osteogenesis assay of SB216763-treated osteoblasts stained by von Kossa (n=5). Scale bar, 50 pm. Part B shows immunostaining of the sections of implants with anti-osterix antibodies (n=6). Scale bar, 50 pm. Part C shows a tube formation assay of SB216763-treated osteoblasts (n=5). Scale bar, 50 pm.
Figure 11 has four parts, A-D, and shows decreased SMADI and increased β-catenin are responsible for the SB216763-induction of osteoblastic-endothelial transdifferentiation. Part A shows expression of SMAD1 or β-catenin in SB216763-treated osteoblasts infected with lentiviral vectors containing CMV promoter-driven SMAD1 cDNA or infected with lentiviral vectors containing β-catenin siRNA. (n=6). Part B shows osteogenesis assay of SB21763-treated osteoblasts infected with lentiviral vectors containing CMV promoter- driven SMAD1 cDNA or infected with lentiviral vectors containing β-catenin siRNA. (n=6). Scale bar, 50 pm. Part C shows immunostaining of sections of implants with anti-osterix antibodies (n=6). Scale bar, 50 pm. Part D shows a tube formation assay of SB21763-treated osteoblasts infected with lentiviral vectors containing CMV promoter-driven SMADI cDNA or containing β-catenin siRNA (n=6).
Figure 12 has four parts, A-D, and shows SB216763 reverses osteogenesis to endothelial differentiation to ameliorate vascular calcification in Mgp-/- mice. Part A shows microCT images of aortic calcification in Mgp-/- and wild type (Mgp*/+) mice after SB216763 treatment (n=6). Scale bar, 5 mm. Part B shows alizarin red staining of aortic tissues (n=6). Scale bar, 5 mm. Part C shows H&E staining of aortic tissues of Mgp-/- and Mgp*/* mice after SB216763 treatment (n=6). Scale bar, 50 pm. Part D shows expression of cbfal and osteopontin (OPN) in aortic tissues of Mgp-/- and Mgp*/* mice after SB216763 treatment (n=8).
Figure 13 has three parts, A-C, and shows specific deletion of GSK33 in osteoblasts reduces vascular calcification. Part A shows microCT images of mice after injection of tamoxifen (n=6). Scale bar, 5 mm. Part B shows H&E staining of aortic tissues of mice after injection of tamoxifen (n=8). Scale bar, 5 mm. Part C shows expression of Cbfal and osteopontin (OPN) in aortic tissues of mice after injection of tamoxifen (n=8).
The examples provided in the instant specification, of generating SB216763-treated osteoblasts infected with lentiviral vectors containing CMV promoter-driven SMAD1 cDNA, or osteoblasts infected with lentiviral vectors containing β-catenin siRNA, the specific gene deletion of GSK3β in osteoblasts, Flkl-eGFP osteoblasts, GFP+CD31+ aortic cells of Osx-GfptgMgp - mice treated with or without SB216763, the study of total aortic calcium of CollalcreERT2GSK3p/loxlxMgp - mice with or without injection of tamoxifen, or of the treatment of various mouse strains in Mgp-/-, wild type (Mgp*/+, Mgp*/*), and the study of cell implants, are not representative or correlative of the broad claims encompassing the ability to treat chronic kidney disease, diabetes mellitus, or fibrodysplasia ossificans progressiva in a subject, or of treating or preventing vascular calcification, inducing or increasing osteoblastic-endothelial transdifferentiation, or inhibiting or decreasing osteogenesis in any subject, as instantly claimed.
In light of the teachings in the art and the specification, one skilled in the art would not accept on its face the examples provided in the instant disclosure as being correlative or representative of the ability to provide treatment effects in a subject. Since the specification fails to provide the requisite guidance for the treatment in any subject, and since determination of the factors required for accomplishing this in any subject is highly unpredictable, it would require undue experimentation to practice the invention over the broad scope claimed.
For these reasons, the instant rejection for lacking enablement over the full scope claimed is proper.
Claims 1, 4, 5, 8, 11-19, 53-56 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 pre-AIA the inventor(s), at the time the application was filed, had possession of the claimed invention.
The breadth of the claims:
The claims are drawn to methods of treating chronic kidney disease, diabetes mellitus, or fibrodysplasia ossificans progressiva in a subject or of treating or preventing vascular calcification, inducing or increasing osteoblastic-endothelial transdifferentiation, or inhibiting or decreasing osteogenesis in a subject in need thereof, comprising administering to the subject any agent that inhibits the activity of or decreases the levels of glycogen synthase kinase 3 (GSK3), which GSK3 is optionally GSK3β, any agent that inhibits the activity of or decreases the levels of mothers against decapentaplegic homolog 1 (SMAD1), or any agent that activates or increases the levels of 3-catenin, which agent optionally comprises any small molecule, any polypeptide, or any inhibitory polynucleotide optionally comprising any siRNA, shRNA, or antisense RNA molecule, or which agent is a polynucleotide encoding β-catenin.
Teachings in the art and in the specification.
The teachings in the art and in the specification are described above in the scope of enablement rejection.
The specification fails to provide the requisite guidance for using the large genus of modulatory agents instantly claimed, and further whereby treatment is provided in any subject. The specification teaches some examples of inhibitory or stimulatory agents, these include SB216763-treated osteoblasts infected with lentiviral vectors containing CMV promoter-driven SMAD1 cDNA or osteoblasts infected with lentiviral vectors containing β-catenin siRNA, specific gene deletion of GSK3β in osteoblasts, Flkl-eGFP osteoblasts, GFP+CD31+ aortic cells of Osx-GfptgMgp - mice treated with or without SB216763, the study of total aortic calcium of CollalcreERT2GSK3p/loxlxMgp - mice with or without injection of tamoxifen. These examples do not provide a representative number of species for the myriad of modulators claimed.
Since the disclosure fails to describe the common attributes and characteristics concisely identifying members of the proposed genus of modulators, and because the claimed genus is highly variant, the description provided is insufficient. One of skill in the art would reasonably conclude that the disclosure fails to provide a representative number of species to describe the broad genus of modulatory agents instantly claimed.
Thus, Applicant was not in possession of the broadly claimed genus.
Claim Rejections - 35 USC § 102
In the event the determination of the status of the application as subject to AIA 35 U.S.C. 102 and 103 (or as subject to pre-AIA 35 U.S.C. 102 and 103) is incorrect, any correction of the statutory basis (i.e., changing from AIA to pre-AIA ) for the rejection will not be considered a new ground of rejection if the prior art relied upon, and the rationale supporting the rejection, would be the same under either status.
The following is a quotation of 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.
Claim(s) 1, 4, 11 is/are rejected under 35 U.S.C. 102(a)(1) as being anticipated by Rawadi et al (WO 2005/027883).
Rawadi et al (WO 2005/027883) teach methods for preventing and or treating bone related diseases in mammals using GSK-3β inhibitors (abstract). Rawadi teach that, in unstimulated cells, GSK-3β phosphorylates the N-terminal domain of β -catenin, targeting it for ubiquitylation and proteasomal degradation. Exposure of cells to Wnts leads to inactivation of GSK- 3β and results in the dephosphorylation of β-catenin, thus escaping ubiquitylation dependent destruction. Unphosphorylated β-catenin accumulates in the cytoplasm and translocates to the nucleus, where it binds to transcription factors and activates transcription of target genes (page 2). Rawadi provides an in vitro model using a pluripotent mesenchymal cell line with the ability to differentiate into osteoblasts when triggered and monitoring osteoblast differentiation markers. An in vivo model was taught using LRP5 knockout animals showing osteopenia phenotype resulting from the absence of the Wnt canonical signaling pathway that involves GSK-3β. This model allowed testing the effect of GSK-3β inhibitors on bone mass (pages 12, 13) See also claims 1-4, 8, 9).
Claim(s) 1, 4, 5, 8, 11, 12, 14, 19 is/are rejected under 35 U.S.C. 102(a)(1) as being anticipated by Y. Yao (Switch of Osteogenesis in Vascular Calcification, Genome, National Institute of Health, pages 1, 2 (2015)).
Y. Yao (Switch of Osteogenesis in Vascular Calcification, Genome, National Institute of Health, pages 1, 2 (2015)) teach that SB216763 is a GSK3 inhibitor that converts osteoblasts into endothelial like cells. Yao teaches that SB216763 or otherwise limiting GSK3 modulates protein levels of SMAD1 and catenin and their transcriptional activity to switch the osteoblastic fate for endothelial differentiation
Yao teaches that SB216763 reduces EC origin osteogenic differentiation and decreases calcification in aorta of matrix Gla protein null mice, an established model of vascular calcification. GSK3 inhibition induces osteoblastic endothelial transdifferentiation to ameliorate vascular calcification in subjects with diabetes mellitus (see entire document).
Claim Rejections - 35 USC § 103
In the event the determination of the status of the application as subject to AIA 35 U.S.C. 102 and 103 (or as subject to pre-AIA 35 U.S.C. 102 and 103) is incorrect, any correction of the statutory basis (i.e., changing from AIA to pre-AIA ) for the rejection will not be considered a new ground of rejection if the prior art relied upon, and the rationale supporting the rejection, would be the same under either status.
The following is a quotation of 35 U.S.C. 103 which forms the basis for all obviousness rejections set forth in this Office action:
A patent for a claimed invention may not be obtained, notwithstanding that the claimed invention is not identically disclosed as set forth in section 102, if the differences between the claimed invention and the prior art are such that the claimed invention as a whole would have been obvious before the effective filing date of the claimed invention to a person having ordinary skill in the art to which the claimed invention pertains. Patentability shall not be negated by the manner in which the invention was made.
This application currently names joint inventors. In considering patentability of the claims the examiner presumes that the subject matter of the various claims was commonly owned as of the effective filing date of the claimed invention(s) absent any evidence to the contrary. Applicant is advised of the obligation under 37 CFR 1.56 to point out the inventor and effective filing dates of each claim that was not commonly owned as of the effective filing date of the later invention in order for the examiner to consider the applicability of 35 U.S.C. 102(b)(2)(C) for any potential 35 U.S.C. 102(a)(2) prior art against the later invention.
Claim(s) 1, 4, 5, 8, 11-19 is/are rejected under 35 U.S.C. 103 as being unpatentable over Rawadi et al (WO 2005/027883) and Y. Yao (Switch of Osteogenesis in Vascular Calcification, Genome, National Institute of Health, pages 1, 2 (2015)), the combination in view of Biver et al (Molecular and Cellular Endocrinology, Vol. 382, pages 120-13 (2014)), Coghlan et al (Chemistry and Biology, Vol. 7, No. 10, pages 793-803 (2000)), Wang et al (Stem Cell Research and Therapy, Vol. 9, pages 1-13 (2018)), and Wang et al (Osteoarthritis and Cartilage, Vol. 19, pages 751-762 (2011)).
The claims are drawn to methods of treating or preventing vascular calcification, inducing or increasing osteoblastic endothelial transdifferentiation, and/or inhibiting or decreasing osteogenesis in a subject comprising administering an agent that inhibits the activity levels of glycogen synthase kinase 3 (GSK3), which GSK3 is optionally GSK3- β, or comprising an agent that inhibits the activity or decreases levels of mothers against decapentaplegic homolog 1 (Smad-1), or which agent activates or increases the levels of β-catenin, or is the β-catenin protein, or which agent optionally comprises the small molecule SB216763, or which agent optionally comprises a polypeptide or inhibitory polynucleotide specific for GSK3 or GSK3- β, which inhibitory polynucleotide optionally comprises, siRNA, shRNA, antisense.
Rawadi et al (WO 2005/027883)(see IDS filed 12-12-22) teach methods for preventing and or treating bone related diseases in mammals using GSK-3β inhibitors (abstract). Rawadi teach that, in unstimulated cells, GSK-3β phosphorylates the N-terminal domain of β -catenin, targeting it for ubiquitylation and proteasomal degradation. Exposure of cells to Wnts leads to inactivation of GSK- 3β and results in the dephosphorylation of β-catenin, thus escaping ubiquitylation dependent destruction. Unphosphorylated βcatenin accumulates in the cytoplasm and translocates to the nucleus, where it binds to transcription factors and activates transcription of target genes (page 2). Rawadi provides an in vitro model using a pluripotent mesenchymal cell line with the ability to differentiate into osteoblasts when triggered and monitoring osteoblast differentiation markers. An in vivo model was taught using LRP5 knockout animals showing osteopenia phenotype resulting from the absence of the Wnt canonical signaling pathway that involves GSK-3β. This model allowed testing the effect of GSK-3β inhibitors on bone mass to be observed (pages 12, 13) See also claims 1-4, 8, 9).
Y. Yao (Switch of Osteogenesis in Vascular Calcification, Genome, National Institute of Health, pages 1, 2 (2015)) (see IDS filed 12-12-22) teach that SB216763 is a GSK3 inhibitor that converts osteoblasts into endothelial like cells. Yao teaches that SB216763 or otherwise limiting GSK3 modulates protein levels of SMAD1 and catenin and their transcriptional activity to switch the osteoblastic fate for endothelial differentiation. Yao teaches that SB216763 reduces EC origin osteogenic differentiation and decreases calcification in aorta of matrix Gla protein null mice, an established model of vascular calcification. GSK3 inhibition induces osteoblastic endothelial transdifferentiation to ameliorate vascular calcification in subjects with diabetes mellitus (see entire document).
Biver et al (Molecular and Cellular Endocrinology, Vol. 382, pages 120-13 (2014)) (see IDS filed 12-12-22) teach the promotion of mesenchymal stem cell (MSC) osteogenic differentiation and the association with increased Smad1 activity. Adult human multipotent mesenchymal stem cells (HMSC) are used for reconstruction of bone defects, fracture repair, and bone tissue regeneration. Factors stimulating mesenchymal cell proliferation include Wnt proteins. Bone morphogenetic proteins (BMPs) are potent inducers of mesenchymal cell differentiation into osteoblasts. BMPs exert their effects through serine/threonine kinase. BMPs activate Smad Type 1 receptors for BMPS phosphorylate SSXS motif at the C terminus of Smad-1, -5, and -8. Phosphorylated Smads form heterocomplexes with Smad-4 and are translocated to the nucleus, where they regulate expression of target genes. The duration of the Smad signal and its translocation to the nucleus are regulated by sequential phosphorylation of GSK3. Biver also teaches the inhibition of GSK3 by SB216753 leads to activation of Smad-1 and catenin- β, in turn regulating osteoblastic HMSC differentiation (see esp. the abstract and introduction on page 120-121).
Coghlan et al (Chemistry and Biology, Vol. 7, No. 10, pages 793-803 (2000)) (see IDS filed 12-12-22) teach SB216763 as an effective inhibitor of GSK-3β, which upon phosphorylation leads to induced transcription of β-catenin in HEK293 cells in vitro (see esp. the text on page 793, Figure 1 on page 794, bridging paragraph, pages 797-798, bridging paragraph, pages 799-800).
Wang et al (Stem Cell Research and Therapy, Vol. 9, pages 1-13 (2018)) (see IDS filed 12-12-22) teach the inhibition of osteoblastic differentiation and mineralization by marrow derived mesenchymal stem cells (MSC-CM). MSC-CM suppressed expression of phosphorylated Smad1/5/8, and induced translocation from the cytoplasm to the nucleus. MSC-CM inhibits beta glycerophosphate and induces vascular calcification through blockade of the BMP2-Smad1/5/8 signaling pathway in vascular smooth muscle cells. Osteo and chondrocytic transformation of vascular smooth muscle cells and their dedifferentiation from a contractile to a proliferative phenotype are important for the initiation and progression of vascular calcification. BMP2-Smad signaling is related to the BMP receptors and Smad transcription factors in osteogenesis. After activation of BMP receptors, the expression and translocation of phosphorylated Smad1/5/8 from the cytoplasm to the nucleus may trigger osteoblast differentiation. After treatment with MSC-CM, the expression of phospho-Smad1/5/8 in the nucleus regressed while cytoplasmic expression increased in VSMCs (see esp. the text on page 1, section entitled MSC-CM blocks BMP2-Smad1/5/8 signaling pathway, pages 4-5, section entitled Noggin, BMP2 antagonist, suppresses VSMC calcification by inhibiting Smad 1/5/8 signaling, pages 5-6).
Wang et al (Osteoarthritis and Cartilage, Vol. 19, pages 751-762 (2011)) (see IDS filed 12-12-22) teach prenatal and postnatal bone formation to be complicated process regulated by multiple growth factors, with β-catenin being an important factor affecting cell fate during early prenatal skeletal development. BMPs, especially BMP-2, and their signaling proteins are required in both chondrogenesis and bone formation at embryonic and postnatal stages. Smad1 is an immediate downstream transducing molecule of the BMP receptor and plays a central role in mediating BMP signaling. Here chondrocyte and osteoblast specific Smad1 cKO mice using the Cre-lopP system was used to investigate the specific role of Smad1 in bond development. Chondrocyte specific deletion of Smad-1 caused ectopic cartilage formation in calvaria at the embryonic stage and led to delayed calvarial bone ossification. Osteoblast specific Smad1 cKO mice were found to have impaired postnatal bone formation (see summary on pages 751-752, Fig. 1 on page 753, Fig. 3 on page 756, Results on page 757, Fig. 4 on page 758)
It would have been obvious to treat vascular calcification, induce or increase osteoblastic endothelial transdifferentiation, and/or inhibit or decrease osteogenesis comprising administering an agent that inhibits the activity levels of glycogen synthase kinase 3 (GSK3), which GSK3 is optionally GSK3- β, or comprising an agent that inhibits the activity or decreases levels of mothers against decapentaplegic homolog 1 (Smad-1), or which agent activates or increases the levels of β-catenin, or is the β-catenin protein, or which agent optionally comprises the small molecule SB216763, or which agent optionally comprises a polypeptide or inhibitory polynucleotide specific for GSK3 or GSK3- β, which inhibitory polynucleotide optionally comprises, siRNA, shRNA, antisense. The teachings of Rawadi, Yao., Biler, Coghlan, Wang and Wang taught examples of the modulatory agents claimed and their effects on osteogenesis and the application of their findings to clinical solutions for bone related diseases.
For these and the aforementioned reasons, the instant invention would have been obvious to one of ordinary skill in the art prior to the effective filing date of the instant invention.
Conclusion
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Jane Zara
1-13-26
/JANE J ZARA/Primary Examiner, Art Unit 1637