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 .
Priority
The instant application, filed 4/26/2023, is the national stage entry of International Application No PCT/US21/62939, filed 12/10/2021, which claims the benefit of priority to U.S. Provisional Patent Application No. 63/123,880, filed on 12/10/2020.
Information Disclosure Statement
The Information Disclosure Statements filed on 06/08/2023 and 02/04/2025, are acknowledged and found to be in compliance with the provisions of 37 CFR § 1.97. Accordingly, the Information Disclosure Statements have been considered.
Application History
Claims 1-20 were originally presented on 06/08/2023. The preliminary amendments to the claims also filed on 06/08/2023 were received and entered. No claims were cancelled or newly added. Claims 1-20 were pending and subject a requirement for restriction, mailed 11/12/2025.
A response to the restriction requirement was filed on 1/12/2026. No additional claims were added.
Accordingly, claims 1-20 are pending.
Election/Restriction
Claims 1-20 were subject to a restriction/election requirement dated 11/12/2025. Applicant’s election of “the invention of group I, claims 1-10 and 20, without traverse, subject to the Rejoinder provisions of MPEP 821.04”, in the reply filed on 1/12/2026 is acknowledged. See Group I defined in the 11/12/2025 Restriction Requirement:
PNG
media_image1.png
219
582
media_image1.png
Greyscale
11/12/2025 Restriction Requirement, page 4.
The footnote from the 11/12/2025 Restriction Requirement is provided below:
PNG
media_image2.png
80
669
media_image2.png
Greyscale
11/12/2025 Restriction Requirement, page 4.
Status of Claims
Claims 1-20 are pending. Claims 11-19 are withdrawn from further consideration pursuant to 37 CFR § 1.142(b), as being drawn to a non-elected invention and species. Therefore, claims 1-10 and 20 read on an elected invention and are therefore under consideration in the instant application.
Claim Rejections - 35 USC § 112(b)
The following is a quotation of 35 U.S.C. 112(b):
(b) CONCLUSION.—The specification shall conclude with one or more claims particularly pointing out and distinctly claiming the subject matter which the inventor or a joint inventor regards as the invention.
The following is a quotation of 35 U.S.C. 112 (pre-AIA ), second paragraph:
The specification shall conclude with one or more claims particularly pointing out and distinctly claiming the subject matter which the applicant regards as his invention.
Claims 1-10 and 20 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.
Claims 1-10 and 20 Indefinite - “natural ERK activating compound”
Claim 1 is drawn to a method of treatment comprising administering “one or more natural ERK activating compounds” as an adjunct to stimulate endogenous stem cells in a patient’s oral cavity, and thus recites a Markush grouping of “natural ERK activating compounds”. The claim term “natural ERK activating compound” is vague, is not defined in the Specification, and as a result, it is not clear which species are covered by the grouping.
While it is not immediately clear from the claims itself, the patient populations are patients with periodontitis (see Specification at 1-2, and at 4, paragraph [00013]).
Reference is made to Saud 2019,1 Sharifi 2020,2 Kornicka 2017,3 and Xue 20184 in this rejection.
Illustrative claim and terms
Claim 1 is illustrative of the issue regarding the claim term “natural ERK activating compound” and is reproduced below. Dependent claims 2-10 and 20 fail to further address which species are covered by the grouping (compare to claim 7, reciting somewhat-specific5 small molecule compounds):
PNG
media_image3.png
270
700
media_image3.png
Greyscale
Claim 1.
To explain, the vague claim term has four parts: “natural”, “ERK”, “activating”, and “compound”.
natural – not defined in the specification, but example compounds are resveratrol, PEITC, “boswellic acids” (no data), and curcumin, each derived from a living organism.
ERK – plausibly synonymous with MAPK (Specification at 3, paragraph [00011]. See also, Specification at 3, paragraph [00010] (“MEK phosphorylation, in turn, activates its kinase activity, and in turn phosphorylates ERK, which is also referred to as MAPK”).
activating - the specification never states what it means by “activating”, in terms of what it activates. It could be ERK, or something else. Moreover, the term “modulating” is used interchangeably with “activating”, see Specification at 6, number 1.10 (defining resveratrol, “boswellic acid”, curcumin and PEITC as “natural ERK modulating compound[s]”) (emphases added).
Further, it is in no way clear if the “activation” or “modulation” must occur in a target cell, or in the surrounding tissue, or systemically in the organism because all forms of administration are permitted (see Specification at 15, paragraphs [00046]-[00047]).
compound – not limited, and thus includes proteins and small molecules.
To visualize some of these signaling pathways, reference is made to Xue 2018. See, e.g., Xu 2018 at 5, Figure 2:
PNG
media_image4.png
745
925
media_image4.png
Greyscale
See, e.g., Xu 2018 at 5, Figure 2.
A working definition for a “natural compound” means any compound that is produced by a living organism. The instant methods of treatment seem to claim every single one of these compounds, so long as they are “ERK activating”. The broadest reasonable interpretation of this means that the “natural compound” activates or modulates anything in the ERK/MAPK pathway, with the level of “activation” or “modulation” left unspecified.
To explain, as the Specification at 3 states, “the ERK pathway … is ubiquitous and the molecular targets may be non-specific”. Therefore, any natural compound may be a “natural ERK activating compound”, so long as it has some “activating” or “modulating” effect on the “ubiquitous” ERK pathway. One of skill in the art would not be able to envision all of the compounds that might qualify as a “natural ERK activating compound” under that interpretation, because any compound might have some “activating” or “modulating” effect on the “ubiquitous” ERK pathway. As a direct result, it not clear which species are covered by the claim. The only compounds excluded from this open set of all compounds are those that are not “natural”.
Moreover, the exemplary compounds Applicant highlights, being “resveratrol”, “curcumin”, and PEITC, have no structural similarity whatsoever as shown below, and no attempts were even made to make a structural activity relationship for these scaffolds, nor actually provide their drawn chemical structures in the specification:
PNG
media_image5.png
368
240
media_image5.png
Greyscale
Curcumin
PNG
media_image6.png
325
196
media_image6.png
Greyscale
Resveratrol
PNG
media_image7.png
249
113
media_image7.png
Greyscale
PEITC
One of skill in the art would not be able to look at the structures of these compounds and envision what it means to be a “natural ERK activating compound”, because of the lack of structural similarity of the exemplified embodiments.
For example, Applicant identifies “boswellic acids” as “natural ERK activating compounds”, but fails to report any data on such compounds or even identify a single one. One of skill in the art at the time of filing would not be able to determine what “boswellic acid” from the entire family of “boswellic acids” might be a “natural ERK activating compound” in the context of the instant claims, because these “boswellic acids” are also entirely structurally distinct from resveratrol, curcumin, and PEITC. See Specification at 13, paragraph [00037]:
PNG
media_image8.png
58
715
media_image8.png
Greyscale
See Specification at 13, paragraph [00037].
Moreover, since the specification never states what it means by “activating” or “modulating”, and fails to provide comparative data of actually administering any compound to patients, or even models of human disease, it is entirely unclear what it means to be “activating” or “modulating” within therapeutic context of these claims.
To highlight this problem, reference is made to a few review articles squarely in the field of art these claims appear to be directed towards. First, see Saud 2019 at 1, shown below. The abstract teaches the use of natural compounds such as icariin, genistein, and resveratrol as phytochemicals for regenerative therapies in mesenchymal stem cells. Further see Saud 2019 at 4, Figure 1, also shown below. It teaches a number a different phytochemicals that have been evaluated in for regenerative therapies in mesenchymal stem cells, including curcumin. According to Specification and the claims, resveratrol and curcumin are “natural ERK activating compounds”. It is entirely unclear what makes these compounds “natural ERK activating compounds”, versus all of the other compounds that state ERK, or have a mechanism of action somewhere along the ubiquitous ERK pathway, such as those shown in Saud 2019 at 5-6, Tables 1 and 2, all shown below. See Saud 2019 at 1:
PNG
media_image9.png
550
778
media_image9.png
Greyscale
Saud 2019 at 1.
See also, Saud 2019 at 4, Figure 1.:
PNG
media_image10.png
844
811
media_image10.png
Greyscale
Saud 2019 at 4, Figure 1.
See also, Saud 2019 at 5, Table 1:
PNG
media_image11.png
505
792
media_image11.png
Greyscale
Saud 2019 at 5, Table 1.
See also, Saud 2019 at 6, Table 2:
PNG
media_image12.png
615
992
media_image12.png
Greyscale
Saud 2019 at 6, Table 2.
Reference is also made to Sharifi 2020, which further reviews phytochemicals on osteogenic differentiation of mesenchymal stem cells. See, e.g., Sharifi 2020 at 877, Figure 2:
PNG
media_image13.png
704
987
media_image13.png
Greyscale
Sharifi 2020 at 877, Figure 2.
It is unclear which of these “natural compounds” are “natural ERK activating compounds” in the context of these claims. For example, “Moringin” is listed as a phytochemical with osteogenic differentiation effect on MSCs. Moringin is a phenyl isothiocyanate, just like PEITC (see claims 6 and 7, structure of Moringin and presumed structure of PEITC provided below).
PNG
media_image14.png
298
175
media_image14.png
Greyscale
Moringin, CAS No. 73255-40-0
PNG
media_image7.png
249
113
media_image7.png
Greyscale
PEITC, CAS No. 2257-09-2
Sharifi 2020 at 879, Table 1, indicates that Moringin increases RUNX-2 and ALP genes in PDLSCs.
PNG
media_image15.png
59
1396
media_image15.png
Greyscale
Sharifi 2020 at 879, Table 1.
It is unclear if that alone makes Moringin a “natural ERK activating compound”, because PDLSCs are dental stem cells in the oral cavity, and modulation of Runx2 and ALP are explicitly considered in claim 5.
Further see Sharifi 2020 at 886, which devotes an entire section to resveratrol in regenerative medicine. Resveratrol affects MAPK, ERK1/2, and a number of related signaling pathways. Given the number of pathways resveratrol affects, it is unclear what makes Resveratrol a “natural ERK activating compound”, and not all of the other “natural compounds” listed in pages 878 to 883 of Sharifi 2020 (not shown herein due to the length).
Reference is also made to Kornicka 2017, which generally reviews the same subject matter. Kornicka 2017 at 952-953, Table 2, lists approximately 33 different plant types, and provides representative “extract[s]” or named compounds that were known at the time of filing to have an effect on mesenchymal stem cells. Under the “Mechanism of action” column, many of these compounds or extracts were known at the time of filing to have an effect on mesenchymal stem cells by interactions with various parts of the ERK pathway, cause “modulation” of factors recited in claim 5, and/or activate/modulate pathways involved in the ERK/MAPK pathways.
As evident from these references, and without a precise definition of what it means to be a “natural ERK activating compound”, it would take an entire team of scientists devoted to a research project of searching for “natural ERK activating compounds” to understand which “natural compounds” activate or modulate anything in the ERK pathway within the context of these claims.
As a result, claims 1-10 and 20 are rejected as indefinite, because it is not clear which species are covered by the grouping of “natural ERK activating compounds”, in view of the vague meaning of the claim term “natural ERK activating compound”.
Claim 5 Indefinite – “can”
Regarding claim 5, the phrase "can modulate the level of expression of one or more of the following" renders the claim indefinite because it is unclear whether the limitation(s) following the phrase are part of the claimed invention. Preferences are properly set forth in the Specification. See MPEP § 2173.05(d).
Accordingly, claim 5 is further rejected as indefinite.
Claims 3, 4, 8, 20 Indefinite – antecedent basis6
Regarding claim 3, it recites the limitation "natural ERK activating". There is insufficient antecedent basis for this limitation in the claim.
Regarding claim 4, it recites the limitation "natural compound". There is insufficient antecedent basis for this limitation in the claim.
Regarding claim 8, it recites the limitation "natural compound". There is insufficient antecedent basis for this limitation in the claim.
Regarding claim 20, it recites the limitation "natural compound". There is insufficient antecedent basis for this limitation in the claim.
Accordingly, claims 3, 4, 8, and 20 are further rejected as indefinite.
Claim 20 Indefinite
Regarding claim 20, which was not discussed in depth above, it is reproduced below and requires a bit of discussion. See claim 20:
PNG
media_image16.png
141
651
media_image16.png
Greyscale
Claim 20.
This claim likely depends from claim 11 as the examiner noted in the 11/12/2025 Restriction Requirement, page 4. Claim 11 has been withdrawn pursuant to the restriction requirement. That claim 20 depends from claim 1 appears to be a typographical error.
To the extent that claim 20 is not canceled, on its face the claim does not make much sense. “A method of treatment …, [wherein the compound is administered] …, wherein [the compound] is selected for further development …”. Once the compound has been administered to the patient, it is unclear how one would select it for further development.
Moreover, the examiner cannot avoid pointing out that the claim explicitly states that the compounds resveratrol, “boswellic acid” (no data reported), curcumin or an extract of curcumin, and PEITC is selected for further development. If the compounds it refers to are not yet developed, it is unclear how the methods of their use are enabled.
Accordingly, claim 20 is further rejected as indefinite, as it is unclear how to select a compound for development, after it has been administered to a patient.
Claim Interpretation
As discussed, claim 1, reproduced below, is drawn to a method of treatment. It has four parts that must be construed to understand the scope of the claim, these being the patient population, the therapeutic endpoint, the active administered, and the route of administration.
A method of treatment of gingiva and/or periodontal ligament tissue to increase dental osteogenesis, mineralization or soft tissue regeneration in the oral cavity, wherein the method comprises administering an effective amount of one or more natural ERK activating compounds to the oral cavity of a patient in need thereof, wherein the patient has existing endogenous stem cells that are present in the oral cavity, and; wherein the natural ERK activating compound is administered to the endogenous stem cells in the oral cavity of the patient.
the patient population
Claim language construed
“a patient in need thereof, wherein the patient has existing endogenous stem cells that are present in the oral cavity, and”
Interpretation
A patient with mild to severe periodontitis
Source
Patients with periodontitis, at “mild disease to severe disease with significant loss of tissue and bone”. See Specification at 2, paragraph [0006].
PNG
media_image17.png
92
696
media_image17.png
Greyscale
and,
“patients with moderate periodontitis”, or “patients with advanced (chronic) disease stage periodontitis”. For the latter, it appears that the claim might inherently require some sort of combination of the compound “in combination with periodontal ligament stem cells”.
See Specification at 4, paragraph [00013]:
PNG
media_image18.png
309
719
media_image18.png
Greyscale
the therapeutic endpoint
Claim language construed
“an effective amount” and “to increase dental osteogenesis, mineralization or soft tissue regeneration in the oral cavity”
Interpretation
“an effective amount” and “to increase dental osteogenesis, mineralization or soft tissue regeneration in the oral cavity”
Source
Plain and ordinary meaning
See also, Specification at 13, paragraph [00033]:
PNG
media_image19.png
92
723
media_image19.png
Greyscale
the active administered
Claim language construed
“natural ERK activating compound”
Interpretation
a “natural” compound that activates or modulates anything in the ERK/MAPK pathway
Further, such compounds “can”, but do not have to, “modulate levels of expression of” cellular alkaline phosphatase (ALP), Runx2, bone marrow stromal cells, CD166, CD90, CD105, Stro-1, ATF4, LRP5, TGFβ, osteopontin (OPN), FAS, FASL, and osteocalcin (OCN). See, e.g., claim 5.
Source
Plain and ordinary meaning
the route of administration
Claim language construed
“wherein the natural ERK activating compound is administered to the endogenous stem cells in the oral cavity of the patient”
Interpretation
wherein the “natural” compound is administered by any route
All forms available, such as systemic (pills), solutions (mouthwashes), compositions (gels), and injections.
Source
See Specification at 15, paragraphs [00046]-[00047]:
PNG
media_image20.png
276
724
media_image20.png
Greyscale
According the examiner interprets claim 1 as reciting:
A method of treatment of gingiva and/or periodontal ligament tissue to increase dental osteogenesis, mineralization or soft tissue regeneration in the oral cavity, wherein the method comprises administering an effective amount of one or more of a “natural” compound that activates or modulates anything in the ERK/MAPK pathway to the oral cavity of a patient with mild to severe periodontitis; wherein the “natural” compound is administered by any route.
Claim Rejections - 35 USC § 102(a)(1) / 35 USC § 103
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.
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.
The factual inquiries for establishing a background for determining obviousness under 35 U.S.C. 103 are summarized as follows:
1. Determining the scope and contents of the prior art.
2. Ascertaining the differences between the prior art and the claims at issue.
3. Resolving the level of ordinary skill in the pertinent art.
4. Considering objective evidence present in the application indicating obviousness or nonobviousness.
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.
Rejection Statements
Claim(s) 1-5 and 8-10 is/are rejected under 35 U.S.C. 102(a)(1) as anticipated by or, in the alternative, under 35 U.S.C. 103 as obvious over Zhang 2018.7
Claim(s) 1-5 and 8-10 is/are rejected under 35 U.S.C. 102(a)(1) as anticipated by or, in the alternative, under 35 U.S.C. 103 as obvious over Han 20178 as evidenced by Wang.9
Claim(s) 1 and 6-7 is/are rejected under 35 U.S.C. 102(a)(1) as anticipated by or, in the alternative, under 35 U.S.C. 103 as obvious over as Forouzanfar 2020.10
Discussion of the Invention
The instant claims are drawn to Applicant’s reports that certain compounds PEITC, resveratrol, and curcumin may stimulate proliferation/differentiation/immunomodulation of gingiva-derived mesenchymal stem cells (GMSCs) and/or periodontal ligament stem cells (PDLSCs) based upon in vitro cell culture studies. In summary, Applicant reports:
PEITC and/or resveratrol promote osteogenesis of PDLSCs (example 2, Specification at 16), purported through the ERK/Wnt pathway.
PEITC and/or resveratrol promote immunomodulation of PDLSCs, examples 3 and 4, Specification at 17.
Curcumin (yellow and/or white) stimulates proliferation of GMSCs and PDLCs, example 5, Specification at 17-18.
Curcumin (yellow) stimulates differentiation of PDLSCs (example 6, specification at 18.
PEITC and/or resveratrol increase GMSC proliferation rate, example 7, specification at 18.
No clinical data were disclosed, and the results are strictly limited to the above brief in vitro characterization.
Background on Dental Stem Cells and Animal Models Periodontitis
Solely in order to guide the reader, reference is made to Egusa 2012,11 Yang 2020,12 and Oz 2011.13
Egusa 2012
Egusa 2012 generally reviews stem cells in dentistry, and provides helpful photographs and tables regarding the inherent characteristics of these types of stem cells. See, e.g., Egusa 2012 at 153, Fig. 2:
PNG
media_image21.png
591
1216
media_image21.png
Greyscale
Egusa 2012 at 153, Fig. 2.
See also, Egusa 2012 at 155, Table 1:
PNG
media_image22.png
755
1239
media_image22.png
Greyscale
Egusa 2012 at 155, Table 1.
Yang 2020
Yang 2020 generally reviews applications of the different types of dental mesenchymal stromal/stem cells (MSCs), and their primary functions in Table 1 at 3 of 24, which is reproduced below. If the reader is not familiar with these types of stem cells, this table further provides the relevant anatomical locations of these cells, and their functions.
PNG
media_image23.png
622
903
media_image23.png
Greyscale
Yang 2020 at 3 of 24, Table 1.
Yang 2020 at 2-5 of 24 further provides helpful background regarding these stem cells. Applicant’s results pertain mainly to PDLSCs and GMSCs, which are discussed on pages 3-5 of Yang 2020.
Yang 2020 at 8-9 of 24 further discusses relevant background on periodontitis, including the use of various animal models of periodontitis such as the miniature swine model, and the use of stem cells as avenues to treat cases of periodontitis. In this discussion Yang 2020 at 8 of 24 teaches that the standard “therapeutic goal of periodontitis is reducing the pocket depth to prevent the progression of the disease and regenerating lost peridontium [133].”
Oz 2011
Oz 2011 discusses various animal models that scientists and clinicians use for studying periodontitis. Oz 2011 at 1-2 provides helpful discussion regarding periodontitis, which is reproduced below. See Oz 2011 at 1-2:
Periodontitis is a highly prevalent, chronic immunoinflammatory disease of the periodontium that results in progressive loss of gingival tissue, the periodontal ligament, and adjacent supporting alveolar bone [1]. In addition to its significant impact on human health, the annual cost of periodontal therapy is estimated to exceed $14 billion in the USA [2]. Furthermore, periodontitis has been associated with systemic diseases, such as cardiovascular complications [3], rheumatoid arthritis [4], and adverse pregnancy outcomes [5].
Chronic inflammation of the periodontium is initiated by complex subgingival biofilms containing several likely periodontal pathogens. The biofilm generally contains a portion of the gram (−) negative anaerobic commensal microbiota as well as opportunistic pathogens of the oral cavity, including Porphyromonas gingivalis (P. gingivalis) [6]. In response to periodontal pathogens, polymorphonuclear cells (PMNs) release destructive reactive oxygen species (ROS), for example, superoxide, via the respiratory burst [7–9], proteinases, and other factors that can damage host tissues [10–12]. These molecules induce further oxidative damage to gingival tissue, periodontal ligaments, and elicit osteoclastic bone resorption [10, 13–15]. The secreted agents also enhance the production of numerous proinflammatory cytokines that contribute to the disease, including interleukin (IL)-1ß, IL-6, and tumor necrosis factor (TNFα), among a broad array of biomolecules that have consistently been reported to be elevated in gingival crevicular fluid (GCF) and tissues of periodontitis patients [16–18], rhesus monkeys [19], and dogs [20]. Levels of these proinflammatory molecules are frequently reduced following periodontal therapy [21, 22].
Because individuals are not equally susceptible to the destructive effects of periodontal infections, periodontitis is not only caused by bacterial infection but also may be associated with host susceptibility [23, 24]. Variability in host responses among individuals contributes significantly to the expression of periodontal diseases [24]. Although human cell cultures were found to be useful models for replicating some aspects of the periodontal disease process at the cellular level, information about the complex host response was not prominent [25]. Thus, research into the host response using animals is critically important in the analysis of periodontal disease and development of improved treatments.
Oz 2011 at 1-2.
Oz 2011 provides the following discussion of the minipig model of periodontitis. These animals naturally develop gingivitis, which may be promoted into a model of periodontitis by placing a ligature in their gums, and/or by inoculating them with the relevant bacterial pathogens. The studies of Zhang 2018 and Han 2017 utilized this model of periodontitis. See Oz 2011 at 2:
1.4. Miniature Pigs
Miniature pigs have oral and maxillofacial structures similar to those of humans in terms of anatomy, physiology, and disease development [37]. The Minnesota miniature pig (minipig) was developed about 60 years ago [38] and has been used extensively in biomedical research [39]. After the age of 6 months, minipigs usually develop gingivitis, manifested by inflamed gingival tissue, accumulated plaque and calculus, and bleeding when probed [37]. There is infiltration of inflammatory cells in the gingival tissue that results in progression to severe periodontal inflammation at 16 months of age with identical histopathology to that seen in humans. Periodontitis in minipigs is promoted in about 4–8 weeks using ligatures, and in association with bacterial inoculations of P. gingivalis, S. mutans, and A. actinomycetemcomitans [37]. Minipigs can be suitable for periodontal as well as orofacial investigations. However, minipigs are relatively expensive, with husbandry issues and few studies to support their use.
Oz 2011 at 2.
Claims 1-5 and 8-10 Anticipated by Zhang 2018
Initial Discussion of Zhang 2018
Zhang 2018 evaluated the local application of the natural compound icariin to enhance periodontal tissue regeneration and relive local inflammation in a minipig model of periodontitis. Icariin is a natural compound that interacts with the “MAPK” pathway.14 Thus, it is a “natural ERK activating compound” in the context of the current invention.
Icariin has the following chemical structure.
PNG
media_image24.png
200
400
media_image24.png
Greyscale
Icariin
The authors assessed icariin for use in periodontal clinical treatment by injecting it into the periodontitis lesions of the minipig model of periodontitis, and reported its effects on periodontal tissue regeneration. See Zhang 2018 at Abstract:
Icariin is the main active ingredient of Epimedii Folium, and it is a promising compound for the enhancement of mesenchymal stem cell function, promotion of bone formation, inhibition of bone resorption, alleviation of inflammation and regulation of immunity. The study investigated the effect of icariin on periodontal tissue regeneration in a minipig model of periodontitis. The minipig model of periodontitis was established. Icariin was injected locally.… Local injection of icariin promoted periodontal tissue regeneration and exerted anti-inflammatory and immunomodulatory function. These results support the application of icariin for the clinical treatment of periodontitis.
Zhang 2018 at Abstract (emphases added).
Zhang 2018 at 1-2 provides relevant background regarding periodontitis, icariin, stem cells and regenerative medicine, placing its study squarely in the area of the art of the instant invention, and teaches the entire inventive concept of the instant claims, see, e.g., excerpts from Zhang 2018 at 1-2:
Mesenchymal stem cell (MSC)-mediated periodontal tissue regeneration is considered an alternative method for periodontitis treatment.4 MSC transplantation for periodontal tissue regeneration has made remarkable strides, but several key problems for its use exist, such as security and ethics.5 There are other alternatives for periodontal tissue regeneration, such as chemical agents, for the treatment of periodontitis. Therefore, there is an urgent need to develop alternative drugs with lower cost and higher efficacy to alleviate inflammation, regulate immunity, and enhance the endogenous functions of MSCs to promote the regeneration of periodontal tissues.
…
Icariin stimulated the proliferation and osteogenic differentiation of human periodontal ligament stem cells (hPDLSCs) in a dose-dependent manner in a suitable concentration range from 0.001 to 1 μg · mL−1, but cytotoxicity limited its use at doses greater than 10 μg · mL−1.13 The researchers also found that the concentration of icariin (0.01–1 μg · mL−1) enhanced the impaired proliferation and osteogenic differentiation potentials of hPDLSCs caused by extracts of Porphyromonas gingivalis, and the most effective concentration of icariin was 0.1 μg · mL−1. Icariin was used in mouse calvarial defect models and senescence models, and the results demonstrated that icariin increased trabecular bone mineral density and improved bone mass via the promotion of bone formation.14 Icariin significantly reduced the inflammatory responses and alleviated the pathological changes in a mouse calvarial osteolysis model.15 The anti-inflammatory and immunoprotective effects of icariin were also observed in immune dysfunctional mice.16
Zhang 2018 at 2 (emphases added).
Zhang 2018 provides clinical evidence that the model was established throughout the Figures, see Fig. 1 at 2,
PNG
media_image25.png
249
546
media_image25.png
Greyscale
and relevant clinical assessments of therapeutic effects of icariin administration at Fig. 2 at 2,
PNG
media_image26.png
127
472
media_image26.png
Greyscale
and provides histopathological assessments of tissue regeneration at Fig. 3 at 3,
PNG
media_image27.png
403
746
media_image27.png
Greyscale
and concludes as follows at 5:
In conclusion, the present study demonstrated that local icariin injection promoted periodontal tissue regeneration and exerted anti-inflammatory and immunomodulatory functions in a minipig model of periodontitis. These results support the potential application of icariin for periodontitis treatment in the future.
Zhang 2018 at 5.
Claim 1 Anticipated by Zhang 2018
Regarding claim 1:
The examiner finds that icariin is a “natural ERK activating compound”. Support is found in Zhang 2018 at 1-2 (“Icariin (ICA) (C33H40O15, molecular weight: 676.67) is a Chinese herbal monomer that is extracted as the main active ingredient of Epimedium.”); MAPK pathway is explicitly discussed in Zhang 2018 at 4-5.
The examiner finds that Zhang 2018 teaches a method of treatment of periodontitis throughout, and the patients in need were the minipigs. The patients were clinically assessed (see Zhang 2018 at 2, Fig. 1 and 2), and were described as “Minipigs with periodontitis” (emphasis added). See Zhang 2018 at 5 under “Establishment of a periodontitis model”. There is no reason to overturn the clinical assessment based on those statements.
The examiner finds that an effective amount is taught. See, e.g., Zhang 2018 at 5 (“0.1 μg · ml−1 icariin (icariin group)”), and at 2 (“These results indicated that local injection of icariin significantly promoted the recovery of periodontal tissues compared to the 0.9% NaCl group in a minipig model of periodontitis.”), at 2, Figure 2.
The examiner finds that the minipigs had existing endogenous stem cells present in the oral cavity, see Zhang 2018 at 4
The candidate concentrations of icariin for periodontal tissue regeneration were investigated. Drug-stimulated periodontal tissue regeneration requires the recruitment and activation of local endogenous MSCs, such as PDLSCs, which differentiate into cementoblasts, periodontal ligament cells, and osteoblasts.4 Our previous study demonstrated that icariin promoted the proliferation and osteogenic differentiation potentials of hPDLSCs in inflammatory conditions at an optimal concentration of 0.1μg·mL−1.13 Therefore, the present study used local injections of 0.1 μg·mL−1 icariin. Clinical assessment, CT scan and histopathological results demonstrated that local icariin injection significantly promoted the regeneration of periodontal tissues, including increased new alveolar bone formation and a more mature and thicker new cementum and typical structure of Sharpey’s fibres in a minipig model of periodontitis. These results suggest that icariin enhanced periodontal tissue regeneration via the promotion of endogenous MSC function, such as proliferation and directed differentiation. Several studies demonstrated that icariin promoted bone formation and inhibited bone resorption in in vivo investigations, which also supports our results that local icariin application enhanced alveolar bone formation in a minipig model of periodontitis.14,20
Zhang 2018 at 4 (emphases added).
The examiner finds that icariin was “administered to the endogenous stem cells in the oral cavity of the patient”. See Zhang 2018 at 5: “Minipigs with periodontitis were injected at three sites surrounding each periodontal defect four weeks after surgery: the distal side of the molar, the mesial side of the molar, and the middle of the molar. Each injection needle was inserted from the mucosa to the surface of the bone (supraperiosteal), and 0.9% NaCl or icariin was injected after significant resistance was encountered. Minipigs with periodontitis were injected with 0.9% NaCl or icariin every two weeks and sacrificed 12 weeks after injection.”
Accordingly, claim 1 is anticipated by Zhang 2018.
Claim 2 Anticipated by Zhang 2018
Regarding claim 2, the examiner finds that the injection site described in Zhang 2018 at 5 (excerpt immediately above) is consistent with administration to PDLSc in the minipigs. The MSCs discussed throughout Zhang 2018 were PDLSCs, which indicates that was the desired target stem cell.
Accordingly, claim 2 is anticipated by Zhang 2018.
Claim 3 Anticipated by Zhang 2018
Regarding claim 3, the examiner finds that the dose used in Zhang 2018 was characterized as the effective amount to increase proliferation of hPDLSCs, and thus it was administered “in an amount effective to increase proliferation of endogenous periodontal ligament (PDL) stem cells … in the patient’s oral cavity”. See Zhang 2018 at 4:
Our previous study demonstrated that icariin promoted the proliferation and osteogenic differentiation potentials of hPDLSCs in inflammatory conditions at an optimal concentration of 0.1μg·mL−1.13 Therefore, the present study used local injections of 0.1 μg·mL−1 icariin. Clinical assessment, CT scan and histopathological results demonstrated that local icariin injection significantly promoted the regeneration of periodontal tissues, including increased new alveolar bone formation and a more mature and thicker new cementum and typical structure of Sharpey’s fibres in a minipig model of periodontitis. These results suggest that icariin enhanced periodontal tissue regeneration via the promotion of endogenous MSC function, such as proliferation and directed differentiation. Several studies demonstrated that icariin promoted bone formation and inhibited bone resorption in in vivo investigations, which also supports our results that local icariin application enhanced alveolar bone formation in a minipig model of periodontitis.14,20
Zhang 2018 at 4.
There is no evidence to suggest that proliferation did not occur.
Accordingly, claim 3 is anticipated by Zhang 2018.
Claim 4 Anticipated by Zhang 2018
Regarding claim 4, MAPK pathway is explicitly discussed in Zhang 2018 at 4-5.
Accordingly, claim 4 is anticipated by Zhang 2018.
Claim 5 Anticipated by Zhang 2018
Regarding claim 5, the examiner finds that icariin can modulate the level of expression of these factors, as evidenced by the discussion in at least pages 4-5 of icariin’s effects on the biological pathways. It would be inconsistent to find otherwise, and the examiner does not need to leave the face of Zhang 2018 to draw this conclusion.
See the title of citation number 22 at page 6 of Zhang 2018: “Dutzan, N. et al. Over-expression of forkhead box P3 and its association with receptor activator of nuclear factor-kappa B ligand, interleukin (IL)-17, IL-10 and transforming growth factor-beta during the progression of chronic periodontitis. J. ClinPeriodontol 36, 396–403 (2009).”
The title of the citation states that forkhead box P3 is associated with receptor activator of transforming growth factor-beta during the progression of chronic periodontitis. Transforming growth factor-beta is recited as “TGFβ” in claim 5.
Now see the text of the citation preceding the citation to 22, in Zhang 2018 at 4 (“Foxp3 also plays a key role in the development and function of Treg cells, which are regulated by IL-1β, and deficiency promoted the inflammatory response.22,33”).
Foxp3 is therefore “forkhead box P3”.
The following sentence in Zhang 2018 at 4 states “It has been reported that icariin promotes the expression of Foxp3, which suggests that icariin exerted its anti-inflammatory ability via regulation of Th17 and Treg cells.34”
If Foxp3 is associated with receptor activator of transforming growth factor-beta during the progression of chronic periodontitis, and icariin promotes the expression of Foxp3, then icariin can modulate transforming growth factor-beta during the progression of chronic periodontitis.
The claim does not require certainty, because Applicant chose to use the word “can” in the claim.
Accordingly, claim 5 is anticipated by Zhang 2018.
Claim 8 Anticipated by Zhang 2018
Regarding claim 8, the examiner finds that the dose used in Zhang 2018 was characterized as the effective amount to increase proliferation of hPDLSCs, and thus the “amount of the natural compound [was] effective to increase the proliferation of endogenous PDL stem cells”.
Accordingly, claim 8 is anticipated by Zhang 2018.
Claim 9 Anticipated by Zhang 2018
Regarding claim 9, Zhang 2018 explicitly teaches the method “wherein the patient is at elevated risk, relative to reference standard, of periodontal ligament or gingiva tissue damage”, by utilizing “Minipigs with periodontitis”, who had significant risk of continuing periodontal ligament and gingiva tissue damage relative to the healthy control minipigs discussed in Zhang 2018 at 5. For example, review the photographs of the “Minipigs with periodontitis” teeth and gums in Figs. 1 and 2,
PNG
media_image28.png
317
669
media_image28.png
Greyscale
PNG
media_image29.png
859
521
media_image29.png
Greyscale
Zhang 2018 at 2, Figs. 1 and 2.
Further see Zhang 2018 at 5, stating that three minipigs without being subjected to the generation of the periodontitis model were used as a control standard:
Six minipigs were used to generate a periodontitis model, and a total of 12 periodontal defects were established. These minipigs were randomly divided into an experimental group and a control group (six periodontal defects in three minipigs in each group). The other three minipigs were used as the healthy control group and sacrificed without any treatment at the end of the study.
Zhang 2018 at 5.
Accordingly, claim 9 is anticipated by Zhang 2018.
Claim 10 Anticipated by Zhang 2018
Regarding claim 10, while GMSCs are not explicitly discussed, the results indicate gingival tissue restoration. See Zhang 2018 at 2 (“The icariin group revealed no obvious red gums in periodontitis lesions and recovery of the gingival margin to an approximately normal level (Fig. 2b).”) (emphases added). See also Zhang 2018 at 2 (“Probing depth (PD) values were (3.72 ± 1.18) mm in the icariin group and (6.56 ± 1.47) mm in the 0.9% NaCl group 12 weeks after surgery.”).
This is consistent with administration to GMSCs according to the Specification at 2, paragraph [0007] (“Without being bound by theory, the treatment with a compound that can modulate a target of the ERK pathway in the oral cavity can, in turn, stimulate periodontal ligament stem cells (PDLSCs) for osteogenesis and/or mineralization, and gingival mesenchymal stem cells (GMSCs) for soft tissue regeneration.”) (emphases added). The specification therefore assigns GMSCs the role of soft tissue regeneration. Moreover, the claim does not require administration directly into the lamina propria. Diffusion to local tissue is explicitly considered. See Specification at 15, paragraph [00046] (“natural ERK modulating compound may be administered by any suitable route, including orally, topically (e.g., on the gums), injection, or implantation.”) (emphases added).
Accordingly, claim 10 is anticipated by Zhang 2018.
Claims 1-5 and 8-10 Obvious over Zhang 2018
The findings of fact and rejections under 35 U.S.C. 102(a)(1) as anticipated by Zhang 2018 are expressly incorporated herein for each claim.
To the extent there that there is a missing claim limitation, or that Applicant may argue for some reason that the instant claims 1-5 and 8-10 are not anticipated by Zhang 2018, the subject matter of these claims were nevertheless obvious over Zhang 2018. As discussed, Zhang 2018 describes the exact experiments and methods that are the subject matter hypothesized by Applicant in the instant claims using a compound from the same class. One of ordinary skill in the art at the time of filing would have a reasonable expectation of success in using the compound icariin as the API in the methods instantly claimed, because Zhang 2018 already provided relevant evidence of its use in validated models of periodontitis. Any differences between the “effective amount” required for certain patient populations represents a result effective variable that one of ordinary skill in the art at the time of filing would have a reasonable expectation of success in tuning utilizing its skills as an ordinary clinician to achieve the desired therapeutic effect, particularly in view of Zhang 2018 at 5 concluding “[t]hese results support the potential application of icariin for periodontitis treatment in the future.”
Accordingly, claims 1-5 and 8-10 were obvious over Zhang 2018.
Claims 1-5 and 8-10 Anticipated by Han 2017 as evidenced by Wang 2016
Initial Discussion of Wang 2016
Wang 2016 is used to show that IGFBP5 is inherently a “natural ERK activating compound”.
See Wang 2016 at 621, Figure 1 (“IGFBP5 activates JNK and MEK/Erk signalling pathways in WJCMSCs and PDLSCs.”); see also Wang 2016 at 622, Figure 2 (“IGFBP5 silencing repressed the JNK and MEK/Erk signalling pathways in WJCMSCs and PDLSCs.”).
Further, see Wang 2016 at 625-626
In conclusion, our results demonstrated that IGFBP5 can lead to activation of JNK and MEK/Erk signalling in the osteogenic differentiation of MSCs. Importantly, the application of specific inhibitors dramatically weakened the IGFBP5-enhanced osteogenic differentiation in MSCs. This study contributed to the molecular mechanisms underlying the osteogenic differentiation of MSCs, showing that IGFBP5 prompted the osteogenic differentiation potentials of MSCs via the JNK and MEK/Erk signalling pathways, revealing the underlying mechanism of IGFBP5-enhanced osteogenic differentiation in MSCs, and identifying a potential target mediator for improving bone tissue regeneration based on MSCs.
Wang 2016 at 625-626.
See also, Wang 2016 at 625, Figure 5:
PNG
media_image30.png
324
941
media_image30.png
Greyscale
Wang 2016 at 625.
Initial Discussion of Han 2017 as evidenced by Wang 2016
Han 2017 evaluated the use of insulin-like growth factor binding protein 5 (IGFBP5), and its recombinant form rhIGFBP5, to enhance periodontal tissue regeneration, proliferation and differentiation in a minipig model of periodontitis.
Han 2017 at 1 provides relevant background regarding periodontitis, IGFBP5, stem cells and regenerative medicine, placing its study squarely in the area of the art of the instant invention.
Han 2017 at 10 as evidenced by Wang 2016 teaches the entire inventive concept of the instant claims:
Improving endogenous MSCs populations and their functions is a key factor in restoring periodontal tissues in periodontitis, especially in the situation without exogenous MSCs application [17, 28]. Application of simple growth factors to recruit and activate the endogenous MSCs is an alternative method. Our previous study revealed that IGFBP5 expression was decreased in PDLSCs and local tissues in periodontitis patients, and the lost expression of IGFBP5 in PDLSCs impaired the osteogenic differentiation potential, which suggested that maintaining IGFBP5 expression might be beneficial for tissue regeneration and inflammation control in periodontitis. Indeed, we found that combined usage of IGFBP5 and exogenous MSCs could promote periodontal tissue regeneration and alleviate local inflammation [24].Thus, in this study, we investigated the possibility of IGFBP5 protein application as a method of treatment for periodontitis independent of exogenous MSCs. First, using TNFα to mimic an inflammatory niche, we discovered that the depletion of IGFBP5 inhibited the migration, chemotaxis, osteogenic differentiation and cell proliferation ability of PDLSCs in the inflammatory condition. These discoveries further confirmed that loss of IGFBP5 impaired MSC function in periodontitis. Furthermore, we confirmed that 0.5 ng/ml rhIGFBP5 enhanced the migration, chemotaxis, osteogenic differentiation and cell proliferation potentials of PDLSCs and rescued the impaired functions of IGFBP5- silenced-PDLSCs in the inflammatory condition. In addition to PDLSCs, BMSCs in alveolar bone are also an alternative endogenous cell resource for periodontitis treatment. Thus, we elucidated that rhIGFBP5 also promoted the migration, chemotaxis, osteogenic differentiation and cell proliferation potentials of BMSCs in the inflammatory condition. Our results indicated that rhIGFBP5 could not only recruit the endogenous MSCs participating in tissue restoration but also promote the proliferation and differentiation capacities of MSCs in the inflammatory niche, suggesting that rhIGFBP5 might be helpful in rescuing the impaired functions of endogenous MSCs.
Han 2017 at 10.
See Han 2017 at 3 of 13 for a summary of the results:
In this study, we investigated the role of IGFBP5 protein in the regulation of MSC function and periodontal tissue regeneration independent of exogenous MSCs in an inflammatory niche. Our results revealed that recombinant human IGFBP5 protein (rhIGFBP5) could activate the migration, chemotaxis, osteo/dentinogenic differentiation and cell proliferation of PDLSCs and bone marrow stem cells (BMSCs) in an inflammatory niche. Additionally, the local injection of rhIGFBP5 restored tissue lesions in periodontitis and had an anti-inflammatory effect in a minipig model of periodontitis. Our results identified a potential cytokine, IGFBP5, for improving tissue regeneration and periodontitis treatment in a manner independent of exogenous MSCs.
Han 2017 at 3 of 13.
The majority the in vivo results are depicted in Han 2017 at 9, Fig. 4, shown below.
PNG
media_image31.png
785
625
media_image31.png
Greyscale
Han 2017 at 9, Fig. 4.
Han 2017 concludes as follows:
In summary, our discovery reveals that rhIGFBP5 can activate the functions of MSCs in an inflammatory niche and that BCOR can form a protein complex with histone demethylase KDM6B, thus regulating IGFBP5 transcriptionally by regulating H3K27me3 methylation on the IGFBP5 promoter. Our findings provide insight into the mechanism underlying the activated capacities of MSCs, identified IGFBP5 as a potential cytokine for improving tissue regeneration and periodontitis treatment independent of exogenous MSCs, and suggest that IGFBP5 has potential application in the dental clinic.
Han 2017 at 12 (emphasis added).
The method was therefore therapeutically successful in treating periodontitis, and Han 2017 explicitly concludes at 12 of 13, that “IGFBP5 has potential application in the dental clinic.”
The data from Han 2017 utilized rhIGFBP5 as the source of IGBFP5 because it was commercially available.15 Whether or not rhIGFBP5 is “natural”, being that it is recombinant, is subject to the meaning of that term. Nevertheless, IGFBP5 itself is natural, and therefore IGFBP5 is a “natural ERK activating compound”, which Han 2017 teaches may be used in the dental clinic.
Further, the study of Han 2017 focused on PDLSCs and bone marrow stromal stem cells (BMSCs) as the stem cell targets receiving administration of rhIGFBP5 (see page 5, right column).
The instant invention appears more drawn to PDLSCs, which are discussed in the main text of Han 2017. However, in case BMSCs become relevant later during examination, the examiner notes that the BMSC discussion in Han 2017 also appears in its supporting information, see Han 2017 at 12 for a description of its contents. The examiner notes mineralization in BMSCs is explicitly discussed:
PNG
media_image32.png
379
571
media_image32.png
Greyscale
Han 2017 at 12.
Claim 1 Anticipated by Han 2017 as evidenced by Wang 2016
Regarding claim 1:
As discussed, the examiner finds that that IGFBP5 is a “natural ERK activating compound”. Support for this inherent feature of IGFBP5 is found in the previously cited sections as evidenced by Wang 2016 at 625, Figure 5, Wang 2016 at 621, Wang 2016 at 622, Figure 2, and Wang 2016 at 625-626.
The examiner finds that Han 2017 teaches a method of treatment of periodontitis throughout, and explicitly states the method may be used with just IGFBP5 in the conclusion at 12 of 13.
The patients in need were the minipigs with periodontitis. See Han 2017 at 4-5 (describing the patients, healthy controls, and clinical assessments). See also, Han 2017 at 9, Fig. 4 (“A minipig model of periodontitis was established”). There is no reason to overturn the clinical assessment based on those statements.
The examiner finds that an effective amount is taught. See, e.g., Han 2017 at 7 (“The areas of periodontal defects were locally injected with either a 0.9% NaCl solution or 0.5 ng/ml rhIGFBP5. At 12 weeks post-injection, the inflammation of gingival tissue was still severe in the 0.9% NaCl group, which accumulated a large amount of calculus and plaque around the marginal gingiva (Fig. 4c). However, in the rhIGFBP5 injection group, the gingival tissue was recovered and similar to the healthy gingiva (Fig. 4d).”) (emphasis added); Han 2017 at 7-10 (text discussion of the achievement of the therapeutic endpoints); and at 9, Fig. 4.
The examiner finds that the minipigs had existing endogenous stem cells present in the oral cavity, see, e.g., Han 2017 at 11:
To further evaluate the effect of rhIGFBP5 on periodontal tissue regeneration, we created a preclinical model of periodontitis by constructing bone defects in minipigs. According to clinical observation, three-dimensional reconstructive CT scans and histopathological photomicrographs, we found that local injection of rhIGFBP5 notably promoted alveolar bone regeneration. Histopathological photomicrographs of the minipig model indicated that local injection of rhIGFBP5 increased periodontal tissues, including new thicker cementum and periodontal ligament which were regenerated in the lesion area, where newly-shaped Sharpey’s fibers inserted into regenerated cementum. Moreover, fewer inflammatory cells had infiltrated the gingival tissue of the defect areas in the rhIGFBP5 treatment group.
Han 2017 at 11 (emphases added).
The examiner finds that rhIGFBP5 was “administered to the endogenous stem cells in the oral cavity of the patient”. See Han 2017 at 4-5.
Postoperative injections were received by minipigs at three areas around each defect: the distal of the molar, the mesial of the molar, and the middle of the molar. Each injection needle was inserted from the mucosa to the surface of the bone (supra periosteal), confronting significant resisting force before the 0.9% NaCl or rhIGFBP5 was injected. For rhIGFBP5 treatment, each area surrounding the defect was injected with 10 uL of 0.5 ng/ml rhIGFBP5, and injection occurred once every 2 weeks.
Han 2017 at 4-5.
Because Han 2017 at 12 teaches that IGFBP5 may be used instead “in the dental clinic”, the examiner finds that Han 2017 teaches that its method may be replaced with IGFBP5, instead of rhIGFBP5.
If there is a question as to whether rhIGFBP5 is a “natural ERK activating compound” due to it being recombinant, the examiner contents that rhIGFBP5 is “natural” as evidenced by it being recombinant and thus being produced by a living organism, but acknowledges that Applicant may contest that interpretation.
Accordingly, claim 1 is anticipated by Han 2017 as evidenced by Wang 2016.
Claim 2 Anticipated by Han 2017 as evidenced by Wang 2016
Regarding claim 2, the examiner finds that the injection site described in Han 2017 at 4-5 is consistent with administration to PDLSc in the minipigs. The primary MSCs discussed throughout Han 2017 were PDLSCs, which indicates that was the desired target stem cell.
Accordingly, claim 2 is anticipated by Han 2017 as evidenced by Wang 2016.
Claim 3 Anticipated by Han 2017 as evidenced by Wang 2016
Regarding claim 3, the examiner finds that the dose used in Han 2017 (see Han 2017 at 7 (“0.5 ng/ml rhIGFBP5”) was characterized as the effective amount to increase proliferation of PDLSc, and thus it was administered “in an amount effective to increase proliferation of endogenous periodontal ligament (PDL) stem cells … in the patient’s oral cavity”. See Han 2017 at 5 (“The Cell Counting Kit-8 assay results showed that 0.5 ng/ml rhIGFBP5 markedly promoted cell proliferation in PDLSCs compared to the control group under TNFα treatment (Fig. 2i).”); see Han 2017 at 7, Fig. 2:
PNG
media_image33.png
802
936
media_image33.png
Greyscale
Han 2017 at 7, Fig. 2.
Accordingly, claim 3 is anticipated by Han 2017 as evidenced by Wang 2016.
Claim 4 Anticipated by Han 2017 as evidenced by Wang 2016
Regarding claim 4, the effects of IGFBP5 on ERK/MAP are inherently taught by Wang 2016, see, e.g., sections referenced in claim 1.
Accordingly, claim 4 is anticipated by Han 2017 as evidenced by Wang 2016.
Claim 5 Anticipated by Han 2017 as evidenced by Wang 2016
Regarding claim 5, modulation of cellular ALP is explicitly discussed in pages 5-8 (see also Figure captions for Figs. 1-3).
Accordingly, claim 5 is anticipated by Han 2017 as evidenced by Wang 2016.
Claim 8 Anticipated by Han 2017 as evidenced by Wang 2016
Regarding claim 8, the examiner finds that the dose used in Han 2017 was characterized as the effective amount to increase proliferation of PDLSc, and thus the “amount of the natural compound [was] effective to increase the proliferation of endogenous PDL stem cells”.
Accordingly, claim 8 is anticipated by Han 2017 as evidenced by Wang 2016.
Claim 9 Anticipated by Han 2017 as evidenced by Wang 2016
Regarding claim 9, Han 2017 explicitly teaches the method “wherein the patient is at elevated risk, relative to reference standard, of periodontal ligament or gingiva tissue damage”, by utilizing minipigs with periodontitis, who had significant risk of continuing periodontal ligament and gingiva tissue damage relative to the healthy control minipigs. See, e.g., Zhang 2018 at 10, Fig. 5 (health = control, healthy minipigs; rhIGFBP5 = minipigs with periodontitis treatment group; NS = minipigs with periodontitis saline group):
PNG
media_image34.png
429
948
media_image34.png
Greyscale
Han 2017 at 10, Fig. 5.
Accordingly, claim 9 is anticipated by Han 2017 as evidenced by Wang 2016.
Claim 10 Anticipated Han 2017 as evidenced by Wang 2016
Regarding claim 10, while GMSCs are not explicitly discussed, the results indicate gingival tissue restoration. See Han 2017 at 5 (“Clinical assessments, including gingival recession (GR), probing depth (PD), and attachment loss (AL), were measured on all experimental area pre-injection and post-injection at 4 weeks and 12 weeks.”) (emphases added). See also Han 2017 at 7 (“At 12 weeks post-injection, the inflammation of gingival tissue was still severe in the 0.9% NaCl group, which accumulated a large amount of calculus and plaque around the marginal gingiva (Fig. 4c). However, in the rhIGFBP5 injection group, the gingival tissue was recovered and similar to the healthy gingiva (Fig. 4d). Clinical assessments of periodontal tissue regeneration were also performed. At 12 weeks post-injection, the PD was 6.6 ± 1.46 mm in the 0.9% NaCl group and 3.2 ± 1.10 mm in the rhIGFBP5 injection group.”).
This is consistent with administration to GMSCs according to the Specification at 2, paragraph [0007] (“Without being bound by theory, the treatment with a compound that can modulate a target of the ERK pathway in the oral cavity can, in turn, stimulate periodontal ligament stem cells (PDLSCs) for osteogenesis and/or mineralization, and gingival mesenchymal stem cells (GMSCs) for soft tissue regeneration.”) (emphases added). The specification therefore assigns GMSCs the role of soft tissue regeneration. Moreover, the claim does not require administration directly into the lamina propria. Diffusion to local tissue is explicitly considered. See Specification at 15, paragraph [00046] (“natural ERK modulating compound may be administered by any suitable route, including orally, topically (e.g., on the gums), injection, or implantation.”) (emphases added).
Accordingly, claim 10 is anticipated by Han 2017 as evidenced by Wang 2016.
Claims 1-5 and 8-10 Obvious over Han 2017 as evidenced by Wang 2016
The findings of fact and rejections under 35 U.S.C. 102(a)(1) as anticipated by Han 2017 as evidenced by Wang 2016 are expressly incorporated herein for each claim.
To the extent there that there is a missing claim limitation, or that Applicant may argue for some reason that the instant claims 1-5 and 8-10 are not anticipated by Han 2017 as evidenced by Wang 2016, the subject matter of these claims were nevertheless obvious over Han 2017 as evidenced by Wang 2016. As discussed, Han 2017 describes the exact experiments and methods that are the subject matter hypothesized by Applicant in the instant claims using a compound from the same class. One of ordinary skill in the art at the time of filing would have a reasonable expectation of success in using the compound IGFBP5 as the API in the methods instantly claimed, because Han 2017 already provided relevant evidence of its use in validated models of periodontitis. Any differences between the “effective amount” required for certain patient populations represents a result effective variable that one of ordinary skill in the art at the time of filing would have a reasonable expectation of success in tuning utilizing its skills as an ordinary clinician to achieve the desired therapeutic effect, particularly in view of Han 2017 at 12 concluding “IGFBP5 has potential application in the dental clinic.”
Accordingly, claims 1-5 and 8-10 were obvious over Han 2017 as evidenced by Wang 2016.
Claims 1 and 6-7 Anticipated by Forouzanfar 2020
As explained in the claim interpretation section, the claims only require:
A method of treatment of gingiva and/or periodontal ligament tissue to increase dental osteogenesis, mineralization or soft tissue regeneration in the oral cavity, wherein the method comprises administering an effective amount of one or more of a “natural” compound that activates or modulates anything in the ERK/MAPK pathway to the oral cavity of a patient with mild to severe periodontitis; wherein the “natural” compound is administered by any route.
An embodiment of claims 1, 6 and 7 is the natural compound curcumin. The claims contemplate all routes of administration, at any dose to effect an increase in dental osteogenesis, mineralization or soft tissue regeneration in the oral cavity.
See Forouzanfar 2020 at 4279, teaching curcumin, at different routes of administration, at different doses, in animal models of periodontitis, with direct effects to increase dental osteogenesis, mineralization or soft tissue regeneration in the oral cavity.
PNG
media_image35.png
707
998
media_image35.png
Greyscale
Forouzanfar 2020 at 4279.
See also, Forouzanfar 2020 at 4280, teaching curcumin, at different routes of administration, at different doses, in clinical trials of periodontitis, with direct effects to increase dental osteogenesis, mineralization or soft tissue regeneration in the oral cavity. While the table header says “gingivitis”, the cited references and text state that the patient populations include patients with periodontitis.
PNG
media_image36.png
885
996
media_image36.png
Greyscale
Forouzanfar 2020 at 4280.
Accordingly, the claims drawn to administration of curcumin by any route, at any dose to effect an increase in dental osteogenesis, mineralization or soft tissue regeneration in the oral cavity, are anticipated by Forouzanfar 2020.
Claims 1 and 6-7 Obvious over Forouzanfar 2020
The findings of fact and rejections under 35 U.S.C. 102(a)(1) as anticipated by Forouzanfar 2020 are expressly incorporated herein for each claim.
To the extent there that there is a missing claim limitation, or that Applicant may argue for some reason that the instant claims 1 and 6-7 are not anticipated by Forouzanfar 2020, for example by arguing that there is not overlap in the patient populations, the subject matter of these claims were nevertheless obvious over Forouzanfar 2020. The claims are too broad by not limiting a route of administration or an exact dose. It would take only ordinary and routine experimentation to prepare a mouthwash or pill comprising an effective amount of curcumin to treat patients with periodontitis in view of the clear clinical results laid out in Forouzanfar 2020.
Accordingly, claims 1 and 6-7 were obvious over Forouzanfar 2020.
Claim Rejections - 35 USC § 103
The following is a quotation of 35 U.S.C. 103 which forms the basis for all obviousness rejections set forth in this Office action:
A patent for a claimed invention may not be obtained, notwithstanding that the claimed invention is not identically disclosed as set forth in section 102, if the differences between the claimed invention and the prior art are such that the claimed invention as a whole would have been obvious before the effective filing date of the claimed invention to a person having ordinary skill in the art to which the claimed invention pertains. Patentability shall not be negated by the manner in which the invention was made.
The factual inquiries for establishing a background for determining obviousness under 35 U.S.C. 103 are summarized as follows:
1. Determining the scope and contents of the prior art.
2. Ascertaining the differences between the prior art and the claims at issue.
3. Resolving the level of ordinary skill in the pertinent art.
4. Considering objective evidence present in the application indicating obviousness or nonobviousness.
This application currently names joint inventors. In considering patentability of the claims the examiner presumes that the subject matter of the various claims was commonly owned as of the effective filing date of the claimed invention(s) absent any evidence to the contrary. Applicant is advised of the obligation under 37 CFR 1.56 to point out the inventor and effective filing dates of each claim that was not commonly owned as of the effective filing date of the later invention in order for the examiner to consider the applicability of 35 U.S.C. 102(b)(2)(C) for any potential 35 U.S.C. 102(a)(2) prior art against the later invention.
Rejection Statement
Claims 1-10 and 20 are rejected under 35 U.S.C. 103 as being unpatentable over Yussif 201616 as evidenced by Yan 2013,17 Wei 2012,18 Pham 2015,19 El-Sayed 2020, 20 and Aytekin 2020,21 in view of Kornicka 2017,22 Petlz 2012,23 Wang 2018,24 Vidoni 2019,25 and Chin 2017,26 in further view of Zhang 2018,27 Han 201728 as evidenced by Wang 2016,29 Linglu 2019,30 and Yang 2020.31
Due to the relative breadth of the claims (multiple targets and pathways), it is appropriate to discuss additional background to guide the reader. The background is incorporated into the following rejections.
Additional Background
As background, reference is made to Linglu 2019, which teaches a comparative analysis of lncRNAs and mRNAs between PDLSCs and GMSCs. In particular, Linglu 2019 at 161 teaches that “highly expressed mRNAs in GMSCs were mainly involved in 49 signaling pathways, such as the PI3K-Akt signaling pathway,” and the “MAPK signaling pathway”. Further, Linglu 2019 teaches that despite having differences in the signaling pathways for GMSCs and PDLSCs, the MAPK pathway stood out as having a significant number of highly expressed genes in both GMSCs and PDLSCs, that the MAPK pathway was known to regulate cellular proliferation, differentiation, and death, and indicated that its core pathways would be relevant for future research. See Linglu 2019 at 163.
The pathway analysis revealed that the differentially expressed mRNAs between PDLSCs and GMSCs were involved in hundreds of cell signaling pathways. After building a pathway relation network, we obtained the crosstalk between these pathways, and some network nodes with larger degrees were thought to be more important regulators, including the MAPK signaling pathway, apoptosis pathway, pathways in cancer, adherence junction pathway, glycolysis pathway and so on. The mitogen-activated protein kinase (MAPK) signaling pathway, for example, is a classical cascade in cells that transmits upstream signals to downstream effectors to regulate cell physiological processes such as proliferation, differentiation and death (Tsai and Nussinov, 2018). Microarray data showed that 12 highly expressed genes in PDLSCs and 13 highly expressed genes in GMSCs were enriched in this pathway, suggesting that these genes might mediate the diverse characteristics of PDLSCs and GMSCs through the MAPK signaling pathway. Similarly, the differentially expressed genes involved in the above core pathways could also be regarded as candidate regulators for future research.
Linglu 2019 at 163.
Reference is made again to Wang 2016, which teaches the MEK/Erk pathway as central to at least osteogenic differentiation in PDLSCs.
See, e.g., Wang 2016 at 625, Figure 5:
PNG
media_image30.png
324
941
media_image30.png
Greyscale
Wang 2016 at 625.
Reference is again made to Yang 2020, Table 1 at 3 of 24, which is reproduced below.
PNG
media_image37.png
732
1063
media_image37.png
Greyscale
Yang 2020 at 3 of 24, Table 1.
Yang 2020 at 3-5 of 24 further provides helpful background regarding the key stem cells applicant studied in its experiments, namely PDLSCs, GMSCs, and discusses SHEDs, which are also relevant hereto. Further relevant to the instant invention is the citation number 67, which teaches in its title: “Endothelial differentiation of SHED requires MEK1/ERK signaling” (emphases added). See Yang 2020 at 5 of 24 and page 18 of 24 for the title.
Claims 1-10 and 20 Obvious over Yussif 2016 as evidenced Yan 2013, Wei 2012, Pham 2015, El-Sayed 2020 and Aytekin 2020, in view of Kornicka 2017, Petlz 2012, Wang 2018, Vidoni 2019 and Chin 2017, in further view of Zhang 2018, Han 2017 as evidenced by Wang 2016, Linglu 2019, and Yang 2020
Ascorbic acid (“AA”), or vitamin C has the following chemical structure:
PNG
media_image38.png
231
188
media_image38.png
Greyscale
Ascorbic acid (“AA”), or Vitamin C
Discussion of Yussif 2016 as evidenced Yan 2013, Wei 2012, Pham 2015, El-Sayed 2020 and Aytekin 2020
A dentist of just ordinary skill at the time of filing, doing his continuing medical education, would come across the paper of Yussif 2016. Little would the ordinary dentist know from reading Yussif 2016 alone, that the clinical method the paper taught was a method of treatment of gingiva and/or periodontal ligament tissue to increase dental osteogenesis, mineralization or soft tissue regeneration in the oral cavity of a patient in need, comprising administering an effective amount of the natural ERK activating compound, being simply vitamin C, or ascorbic acid “AA”, wherein the patient has existing endogenous stem cells that are present in the oral cavity, and wherein the vitamin C is administered to the endogenous stem cells in the oral cavity of the patient.
This ordinary dentist reading Yussif 2016 would find that its authors evaluated the anti-inflammatory effect of local administration of AA/vitamin C for the clinical treatment of persistent gingival inflammation. Yussif 2016 found that this route of administration (local injection) was clinically successful. See Yussif 2016 at Abstract.
PNG
media_image39.png
135
816
media_image39.png
Greyscale
Yussif 2016 at Abstract.
Injection of vitamin C was described as “locally introduced in relation to the keratinized gingival tissues with prevalent extension to the whole target region…”. See Yussif 2016 at 3, where it also describes the concentration used (200-300 mg/1-1.5 mL of AA):
PNG
media_image40.png
281
436
media_image40.png
Greyscale
Yussif 2016 at 3.
Yussif 2016 at 4 provides photographic evidence of the exterior of the patients’ gums to show the reduction of gingival inflammation:
PNG
media_image41.png
549
867
media_image41.png
Greyscale
Yussif 2016 at 4.
Yussif 2016 at 5, Table 1 documents the histopathological assessments of specimens obtained before, during and after treatment with local injections of AA:
PNG
media_image42.png
761
672
media_image42.png
Greyscale
Yussif 2016 at 5, Table 1.
Yussif 2016 at 6 teaches that local injection was preferred for this method of treatment:
The local injection of vit-C was preferred rather than the usage of topical vitamin C gel or dentifrice that was previously used by Daniels and Jefferies [22] and Shimabukuro et al. [23]. Vitamin C is a water-soluble agent that has superficial penetrating effect. Many studies used different forms of lipid soluble topical vitamin C in order to overcome its limited absorption. These studies met several limitations such as very long term improvement time (12 weeks), easily dislodged from the oral tissues, limited absorption, instability when exposed to solutions, air, heat, or light, and localized enamel erosions [9, 24, 25].
Yussif 2016 at 6.
Yussif 2016 at 6-7 discusses the results by characterizing the impact of the local administration on an anti-inflammatory effect, wound healing, and increased resistance to cell death, and teaches the inventive concept of using vitamin C as an adjunct for the nonsurgical treatment of gingival inflammation. It highlights some of the results a true “phenomenon”.
Once inflammation subsided, representative tissue biopsies have been obtained 7 days following the last injection from the patients indicated for gingivoplasty or gingivectomy. Clinically significant reduction of the pseudogingival enlargement was associated with return of the basal keratinocytes to their normal proliferative pattern which is the main role of vitamin C [26, 27]. The anti-inflammatory action of vit-C was also evident and was detected in our specimens as a reduced intraepithelial edema and inflammatory cells.
Other characteristic changes which occurred after vit-C administration were in accordance with Nusgens et al. [28]. An increased number of fibroblasts were clearly detected forming more collagen fibers that showed more maturation and bundles formation following second injection. Numerous newly formed small blood capillaries were also detected. These features are consistent with its known essential role in the formation of new connective tissue in a healing wound. This is because it acts as a cofactor for enzymes critical in collagen formation, the main component of the connective tissue that forms the framework around which the new tissue is rebuilt. Collagen also represents an essential component in the wall of blood vessels. This is why, despite the increase in number of blood vessels, redness and bleeding tendency markedly decreased clinically. These vessels provide more nutritional and oxygen support to chronically irritated and continuously damaged mucosal areas, improving their healing.
Our results were in accordance with Boyce et al. They detected that vitamin C promotes the development of an organization of the basement membrane and also restores the epidermal barrier within 2-3 weeks. Furthermore, it promotes the wound closure and reduces its contraction which limits the incidence of scare formation [27].
Histologically, a characteristic cellular vacuolization was observed in all groups. In the preoperative specimens, vacuolated cells appeared as clear cells with small pyknotic dark nucleus representing signs of degeneration. They were found in clusters widely distributed throughout all epithelial layers. In contrast, vit-C associated vacuolated cells were individually distributed along the epithelial layers with greatest aggregations in basal cells. The nuclei were rounded or kidney shaped with nearly normal stains. A similar vacuolization was found in immediate and 15-minute biopsies excised after intradermal injection of local anesthesia described by Kimura et al. [29]. They attributed this phenomenon to the injection procedure itself rather than the used solution spatially with the presence of vacuolization in biopsies. However, the presence of these cells in after-week biopsies may be due to the ability of vitamin C to increase the cell resistance to death. Furthermore, the basal localization of vacuolated cells, adjacent to injection, may indicate proper infusibility of vit-C.
All these positive clinical changes were met by further patients’ cooperation and resulted in their end-treatment satisfaction.
These data suggest a significant enhancement of the gingival health by the usage of the antioxidant approach. Finally, we recommend the usage of the intraepidermal vitamin C injection as an adjunctive approach for the conventional nonsurgical treatment modality. Further studies with long term follow-up are recommended.
Yussif 2016 at 6-7 (emphases added).
As evidenced by Yan 2013, vitamin C is a natural compound that inherently activates ERK in PDLSCs:
Yan 2013
See, e.g., Yan 2013 at Abstract:
The differentiation of periodontal ligament (PDL) progenitor cells is important for maintaining the homeostasis of PDL tissue and alveolar bone. Vitamin C (VC), a water-soluble nutrient that cannot be biosynthesized by humans, is vital for mesenchymal stem cells differentiation and plays an important role in bone remodeling. Therefore, the objective of this study was to determine the function and mechanism of VC in PDL progenitor cells osteogenic differentiation at the molecular level. We demonstrated that VC could induce the osteogenic differentiation and maturation of PDL progenitor cell without other osteogenic agents. During the process, VC preferentially activated ERK1/2 but did not affect JNK or p38. Co-treatment with ERK inhibitor effectively decreased the Vitamin C-induced expression of Runx2. ERK inhibitor also abrogated Vitamin C-induced the minimized nodules formation. PELP1, a nuclear receptor co-regulator, was up-regulated under VC treatment. PELP1 knockdown inhibited ERK phosphorylation. The overexpression of PELP1 had a positive relationship with Runx2 expression. Taken together, we could make a conclude that VC induces the osteogenic differentiation of PDL progenitor cells via PELP1-ERK axis. Our finding implies that VC may have a potential in the regeneration medicine and application to periodontitis treatment.
Yan 2013 at Abstract (emphases added).
Further, as evidenced by Wei 2012, Vitamin C inherently stimulates PDLSCs proliferation:
Wei 2012
Wei 2012 at 3219 teaches that AA/Vitamin C (abbreviated at Vc) inherently stimulates PDLC proliferation at an optimal concentration of 20.0 ug/mL. See also, Wei 2012 at 3220, Fig. 3:
PNG
media_image43.png
697
836
media_image43.png
Greyscale
Wei 2012 at 3220, Fig. 3.
Moreover, as evidenced by Pham 2015, vitamin C inherently stimulates GMSC proliferation:
Pham 2015
Pham 2015 at 220, Figs. 3 and 4, shows that AA inherently stimulates GMSC proliferation at 10 to 300 μM concentrations, peak at 250 μM (GMSCs are referred to as GSCs in Pham 2015). See, e.g., Pham 2015 at 221, Figure 3a:
PNG
media_image44.png
490
1216
media_image44.png
Greyscale
Pham 2015 at 221, Figure 3a.
Further, as evidenced by El-Sayed 2020, Vitamin C is a natural ERK activating compound in GMSCs:
El-Sayed 2020
El-Sayed 2020 evaluated the use of ascorbic acid, i.e., vitamin C, alone and in combination with inflammatory cytokines “on multipotency/pluripotency, proliferative, and differentiation characteristics of G-MSCs.” See El-Sayed 2020 at Abstract. El-Sayed 2020 uses the acronym G-MSC, rather than the GMSC abbreviation, for Gingival mesenchymal stem/progenitor cells.
El-Sayed 2020 at 1-2 provides relevant background regarding periodontitis, ascorbic acid stem cells (GMSCs in particular) and regenerative medicine, placing its study squarely in the area of the art of the instant invention. Wnt/β-catenin pathway activation by AA is explicitly discussed throughout (see, e.g., El-Sayed 2020 at 6-7, Fig. 2); ERK1/2 phosphorylation explicitly discussed at page 8 (right column). Because AA is a natural compound, and it activates the Wnt/β-catenin pathway, it is a “natural ERK activating compound” within the context of the current invention.32
El-Sayed 2020 found in the inflammatory environment, AA stimulation of GMSCs leads to a burst of proliferation which is then attenuated over a period of days in an inflammatory environment. 250 μg/ml AA was used in the studies (see El-Sayed 2020 right column, no 2.4), further see, e.g., El-Sayed 2020 at 5, Figures 2(b) and 2(c), excerpts below, as well as Table 2 at page 8 (table 2 is too lengthy to insert herein).
PNG
media_image45.png
288
514
media_image45.png
Greyscale
PNG
media_image46.png
690
271
media_image46.png
Greyscale
PNG
media_image47.png
609
269
media_image47.png
Greyscale
El-Sayed 2020 at 5, Figures 2(b) and 2(c).
El-Sayed 2020 at 12 proposes the following regenerative method of treatment
PNG
media_image48.png
298
379
media_image48.png
Greyscale
El-Sayed 2020 at 12.
Moreover, as evidenced by Aytekin 2020, it reinterprets the results of Yussif 2016 by studying the local application of vitamin C at doses commensurate with those used to target PDLSCs and GMSCs:
Aytekin 2020
Aytekin 2020 evaluated the local application of the natural compound ascorbic acid (“AA”)/Vitamin C into the subperiosteum of the buccal gingiva of the Sprague-Dawley albino rat model of periodontitis. See Aytekin 2020 at Abstract.
PNG
media_image49.png
556
1020
media_image49.png
Greyscale
Aytekin 2020 at Abstract.
Aytekin 2020 at 1-2 provides relevant background regarding periodontitis, vitamin C, and cites studies of vitamin C’s application in stem cells and regenerative medicine (see citation no. 22; see also citation no. 48, which is discussed at page 430, pertaining to AA’s use “in promoting osteoblastic differentiation in [PDLSCs]”), placing its study squarely in the area of the art of the instant invention.
See Aytekin 2020 at 426 for a discussion of the model of periodontitis and the dose of AA:
PNG
media_image50.png
348
633
media_image50.png
Greyscale
PNG
media_image51.png
349
633
media_image51.png
Greyscale
Aytekin 2020 at 426.
The therapeutic endpoint of the study was alveolar bone loss and attachment, and further evaluated its effects in the gingival tissues. See, e.g., Aytekin at 429-430, Figs. 4 and 5, and Table 1, excerpts provided below.
PNG
media_image52.png
267
651
media_image52.png
Greyscale
PNG
media_image53.png
397
655
media_image53.png
Greyscale
PNG
media_image54.png
380
1269
media_image54.png
Greyscale
PNG
media_image55.png
217
635
media_image55.png
Greyscale
PNG
media_image56.png
655
641
media_image56.png
Greyscale
Aytekin at 429-430, Figs. 4 and 5, and Table 1.
Aytekin 2020 at 430 cites Yussif 2016, which it interprets as follows:
In addition, Yussif et al. [51] applied local vitamin C treatment after phase 1 periodontal therapy. Their clinical and histological findings have shown that vitamin C is an effective adjunctive treatment in reducing various degrees of chronic gingival inflammation. We may think local vitamin C therapy minimized alveolar bone and periodontal ligament destruction.
Aytekin 2020 at 430 (emphasis added).
Aytekin 2020 at 430 concedes the issues with the particular model of periodontitis used in its own studies, and concludes as follows:
We preferred the local application path, unlike previous vitamin C studies. Advantages of local applications include the conversion of low doses of the drug into high concentrations at the related region, less administration, less systemic adverse effects and higher patient acceptance than systemic drugs [52].
A potential limitation of this study is the lack of dose dependent drug groups. Studies evaluating the therapeutic effect of vitamin C on alveolar bone loss in periodontitis are needed to determine the appropriate dose for in vitro experiments. Another limitation of our study is that the use of rats having ability to produce their vitamin C may not be the best model to monitor vitamin C as a treatment method. Furthermore, the ligature model is an additional limitation. This model induces acute inflammation, which is not directly equivalent to that observed in chronic periodontitis in humans. Also, considering the fact that this was based on an experimental animal model, the administered dose of vitamin C and findings of this study cannot adapt directly to humans. Furthermore, repeated local injections of vitamin C are not suitable for clinical use in patients. Additional studies are needed to shed light on the effects of local administration of vitamin C in term of different doses, time intervals and local release agents as route of administration.
Aytekin 2020 at 430.
Claims 1-10 and 20 were Obvious at the Time of Filing
Accordingly, while it might not have been clear to the dentist of just ordinary skill in the art at the time of filing, doing his continuing medical education, the Aytekin 2020 paper, which cites Yussif 2016, and reinterprets its results, teaches that Yussif 2016 as evidenced by Yan 2013, Wei 2012, Pham 2015, El-Sayed 2020, and Aytekin 2020 indeed teaches a clinical method of treatment of gingiva and/or periodontal ligament tissue to increase dental osteogenesis, mineralization or soft tissue regeneration in the oral cavity of a patient in need, comprising administering an effective amount of the natural ERK activating compound, being simply vitamin C, wherein the patient has existing endogenous stem cells that are present in the oral cavity, and wherein the vitamin C is administered to the endogenous stem cells in the oral cavity of the patient.
Aytekin 2020 essentially repeats the clinical studies of Yussif 2016, using a rat model of periodontitis. As discussed above, Aytekin 2020 utilized doses of AA at 250 μg/ml AA, which appears to be far lower than Yussif 2016’s dose. As evidenced by Pham 2015, the 250 μg/ml AA was optimal to stimulate GMSC proliferation. Moreover, as evidenced by El-Sayed 2020 at 7, this dose results in a burst of GMSC proliferation even in the inflammatory microenvironment, so long as the inflammatory microenvironment had not drove the GMSCs into self-senescence. Moreover, the therapeutic endpoint of Aytekin 2020 showed promising results to the periodontal ligament tissue, with respect to alveolar bone loss and attachment loss.
Further, as evident in the histopathological assessments and the discussion of the results in Yussif 2016, its method resulted in substantial repair and remodeling of the gingival tissue. Indeed, Yussif 2016 at 7 characterizes some of the results as a “phenomenon” (emphasis added).
Further, patients with periodontitis were excluded from the study (see Yussif 2016 at 3, Figure 2), indicating that the patients had existing endogenous stem cells in their oral cavity (to be consistent with the instant Specification, see Specification at 4, paragraph [00013], which discusses that patients with chronic periodontitis may lack such endogenous stem cells).
Further, Yussif 2016 at 6 teaches that local injection of vitamin C is preferred, in view of it being “a water-soluble agent that has superficial penetrating effect.” Moreover, the doses of AA were high – (200-300 mg/1-1.5 mL of AA). Whether or not this dosage had an effect on the stem cells present in the patients’ oral cavities is however unclear (see, e.g., Pham 2015 and Wei 2012, which show that the effects of AA on GMSCs and PDLSCs in vitro taper past a certain concentration).
Further, as clear from the local administration sites of both of the studies – Yussif 2016 at 3 (“locally introduced in relation to the keratinized gingival tissues with prevalent extension to the whole target region…”), Aytekin 2020 at 426 (“administered into the subperiosteum of the buccal gingiva of the right mandibular first molar teeth for three times in intervals of 2 days”) – clinicians of ordinary skill in the art at the time of filing had understood how to administer by local injection a solution of a “natural ERK activating compound” to achieve therapeutic effects, particularly with respect to the gingiva and periodontal ligament tissues.
However, Aytekin 2020 concedes the limitations of the particular rat model used. Aytekin 2020 indicates that it would be therapeutically successful in humans but for the requirement of multiple injections. See Aytekin 2020 at 430 (“Furthermore, repeated local injections of vitamin C are not suitable for clinical use in patients. Additional studies are needed to shed light on the effects of local administration of vitamin C in term of different doses, time intervals and local release agents as route of administration.”).
Well, repeated injections sounds painful, and it seems logical that the dentist of ordinary skill in the art at the time of filing may want to avoid hurting his patients. Thus the methods of treatment of gingiva and/or periodontal ligament tissue to increase dental osteogenesis, mineralization or soft tissue regeneration in the oral cavity of Yussif 2016 as evidenced by Yan 2013, Wei 2012, Pham 2015, El-Sayed 2020, and Aytekin 2020, comprising administering an effective amount of the natural ERK activating compound AA/vitamin C, wherein the patient has existing endogenous stem cells that are present in the oral cavity, and wherein the vitamin C is administered to the endogenous stem cells in the oral cavity of the patient, were ready for improvement.
It would be reasonable for such an ordinary dentist to seek to add an additional agent to the AA cocktail administered to his patients’ in need oral cavities, in order to boost the activity of the cocktail administered, because, and in further view of Zhang 2018, and Han 2017 as evidenced by Wang 2016, the discussion of which in the previous 102/103 section are fully incorporated herein, clinicians of ordinary skill in the art at the time of filing had already discovered the therapeutic effects when a “natural ERK activating compound” was administered locally to a patient’s endogenous stem cells, particular for patients with periodontitis.
It would be reasonable to therefore expect the dentist of ordinary skill in the art at the time of filing to turn to reviews of stem cell literature and find papers like Kornicka 2017 that exemplified the use of phytochemicals and plant extracts for stimulating the proliferation and differentiation of stem cells.
Kornicka 2017, for example at 952, Table 2, teaches that such natural substances are “highly available”, “non-toxic”, “cost-effective”, and have a “wide spectrum of application”, in stark contrast to the “Synthetic stimulators” that are “usually toxic in high doses”, cause “side effects during prolonged application”, and have “high costs of manufacturing”:
PNG
media_image57.png
407
906
media_image57.png
Greyscale
Kornicka 2017 at 952, Table 2.
Not wanting to hurt his patients, it would be obvious for the ordinary dentist to select a “non-toxic” natural alternative over a “usually toxic in high dose” synthetic alternative.
Further, the majority of the compounds listed in Kornicka 2017 alone have a mechanism of action on the MAPK pathway, so the ordinary dentist may be reasonably drawn to those. While these references may be overwhelming to the ordinary dentist, in view of AA/vitamin C stimulating these mechanisms in both GMSCs and PDLSCs through the same pathway (see, e.g., El-Sayed 2020 and Yan 2013 discussion above), the ordinary dentist may be further drawn to these compounds and perform a quick side review of the literature, wherein he would find that each one provided a reasonable expectation of success when administered to his patients in need, because
Yang 2020 at citation 67 teaches that Endothelial differentiation of SHEDs require MEK1/ERK signaling,
Wang 2016 at 625 teaches that MEK/Erk pathway as central to at least osteogenic differentiation in PDLSCs, and
Linglu 2019 at 163 teaches that the MAPK pathway is highly expressed in both GMSCs and PDLSCs, and that its role was “to regulate cell physiological processes such as proliferation, differentiation and death”.
Thus, the ordinary dentist may be further drawn to such compounds that interact with the MAPK pathway, had he reviewed the literature, which he could confirm by seeing the entire discussion of Icariin on page 954-955 of Kornicka 2017, and the results of Zhang 2018.
Returning to Kornicka 2017, the dentist of ordinary skill in the art at the time of filing would find that the very first compound discussed at length in Kornicka 2017 is resveratrol. See Kornicka 2017 at 952 and 954. It would be reasonable for the ordinary dentist to start there because it was discussed at length, and further review the literature on resveratrol in stem cells, so that he could begin improving his cocktail to administer to his patients.
Regarding effective doses, this ordinary dentist would learn from Kornicka 2017 that “[t]he effect of resveratrol on osteogenic differentiation of hMSCs was shown to be a dose dependent as Peltz et al. (Peltz et al., 2012) observed maximum enhancing effect at 0.1 μM, which gradually decreased at higher concentrations.”).33 From there this ordinary dentist would learn that: “Interestingly, cell proliferation rate based on EdU assay was also significantly stimulated by 5 and 10 μM resveratrol.” Peltz 2012 at 8. Indeed, Peltz 2012 taught him that he could push the concentration of resveratrol up stepwise from 0.1 to 1, to 5 and then 10 μM, because “Resveratrol increased cell proliferation rate in a time and dosage dependent manner”, see Peltz 2012 at 4-5 and at 8, Figure 6. Wanting to make his cocktail more potent, he might test each of these concentrations, through his ordinary and routine experimentation using his skills as just an ordinary dentist at the time of filing, and have a reasonable expectation of success that Resveratrol would increase cell proliferation rate in a time and dosage dependent manner.
However, and seeking to find the effects specific to GMSCs and PDLSCs, the ordinary dentist would find that Vidoni 2019 and Wang 2018 had independently discovered that resveratrol combined with ascorbic acid stimulates osteogenic differentiation of GMSCs and PDLSCs.
For example, see Vidoni 2019 at 5, Fig. 2, where the ordinary dentist would find the identical concentration of resveratrol used that Applicant reports in the same GMSCs in the Specification at 18-19, just now mixed with “50 μg/mL of L-ascorbic acid 2-phosphate …, 100 nM of dexamethasone …, and 10 mM of β-glycerophosphate ….” Vidoni 2019 at 2 of 17.
See, e.g., Vidoni 2019 at 5, Fig. 2, reproduced below.
PNG
media_image58.png
679
705
media_image58.png
Greyscale
Vidoni 2019 at 5, Fig. 2.
See also, Vidoni 2019 at 5 (“Representative images in Fig. 2 (and relative quantification) show that the osteogenic differentiation promoted by RV is maximal at 1 μM and declines when concentration raises up to 100 μM, which turned out to be toxic. When 1 μM RV was added to the differentiation medium, osteogenic differentiation of HGMSCs (as mirrored by the mineralization of the extracellular matrix) was accelerated, indicating a synergism between RV and osteogenic differentiation factors (Fig. 3).”) (emphases added).
Further, the ordinary dentist would find that Wang 2018 at 3 reported results in PDLSCs derived from healthy patients and patients with periodontitis, and assessed in 10 nM resveratrol mixed with “50 μg/ml ascorbic acid …, 2mM β-glycerophosphate…, and 10 nM dexamethasone….” In the studies Wang 2018 also tested inflammatory conditions similar to those within the desired patient population. See Wang 2018 at 3 (“For TNF-α treatment, 10 ng/ml human recombinant TNF-α … was added into the osteogenic-inducing media.”). From Wang 2018, the ordinary dentist would learn of the following remarkable results:
See Wang 2018 at 7-12, Figures (figure caption titles excerpted):
Fig. 2 Resveratrol (RSV) rescued the cell aggregate formation and osteogenic differentiation of N-PDLSCs under inflammatory cytokine tumor necrosis factor alpha (TNF-α) treatment.
Fig. 3 RSV treatment improved the osteogenic and bone regenerative potential of P-PDLSCs.
Fig. 4 RSV treatment promoted the cell aggregate formation of P-PDLSCs.
Fig. 5 RSV treatment facilitated the alveolar bone regeneration of P-PDLSC aggregates in a rat periodontal defect model
Wang 2018 at Figures on 7-12 (figure caption titles).
Compiling his notes, the ordinary dentist would learn that the following effective amounts were already taught for resveratrol in mesenchymal stem cells:
0.1 μM, Peltz et al., 2012, with resveratrol increased cell proliferation rate in a time and dosage dependent manner from 0.1, 1, 5, and 10 μM,
1 μM to 100 μM for GMCSs mixed with 50 μg/mL AA and other differentiation factors, Vidoni 2019, optimal at 1 μM for differentiation, 5 μM and 10 μM also taught, but cytotoxic beyond 100 μM, and
10 nM, or 0.01 μM in PDLSCs, mixed with 50 μg/mL AA and other differentiation factors, also tested under inflammatory conditions, Wang 2018.
Well, the ordinary dentist would be targeting his patients endogenous stem cells, by following the procedures of Yussif 2016, therefore he would likely not need to inject the other differentiation factors. But the combination with AA may be desirable, as indicated by the exemplary results of Yussif 2016 and Aytekin 2020.
Armed with this information, the ordinary dentist would check if resveratrol had been used in methods of treating periodontitis, and come across papers like Chin 2017, that studied resveratrol and its derivatives to treat periodontitis, see, e.g., Chin 2017 at 105, Figure 1:
PNG
media_image59.png
633
653
media_image59.png
Greyscale
Chin 2017 at 105, Figure 1, and learn from the paper that resveratrol was expected to increase cellular proliferation as well combat inflammation driven by the bacteria mainly responsible for his patients’ periodontitis, see, e.g., Chin 2017 at 106.
Moreover, the ordinary dentist would learn that treatment with resveratrol, curcumin, or their combination had already been evaluated in models of periodontitis, and clinical trials were ongoing to assess resveratrol supplementation as means of treating periodontitis. See Chin 2017 at 103-4.
Periodontitis was treated with either resveratrol, curcumin, or their combination.44–47 Intergroup comparisons of morphometric outcomes showed higher bone-loss values in the placebo group (P < 0.05) compared with the resveratrol, curcumin, and combined groups. There was no difference in bone-loss values among the resveratrol, curcumin, and the combined groups (P > 0.05). Immuno-enzymatic assays of gingival tissues showed a lower concentration of IL-1β in the combined group compared with the placebo group (P < 0.05). However, higher detected IL-4 levels were shown in the resveratrol, curcumin, and combined groups compared with the placebo group (P < 0.05). Only resveratrol caused a reduction in IFN-γ levels (P < 0.05). There was no difference in TNF-a levels observed among the four groups (P > 0.05). Besides, resveratrol and curcumin were able to reduce loss of the alveolar bone in an animal model of periodontitis when agents were added singly or in combination, but the effects were neither synergistic nor additive.44 Furthermore, the continuous systemic administration of resveratrol also decreased ligature-induced peri-odontal breakdown.10,46,47 Those studies and our results10 confirmed the positive effects of resveratrol on a ligature-induced periodontitis animal model. Recently, a human clinical trial reported by Zare Javid et al. suggested that resveratrol supplementation may be beneficial as adjuvant therapy along with nonsurgical periodontal treatment in insulin resistance and improving the periodontal status in patients with diabetes and periodontal disease.48
Chin 2017 at 103-4.
Accordingly, the dentist of just ordinary skill in the art at the time of filing would have a reasonable expectation that combining ascorbic acid, with at least resveratrol, in an effective amount of each, would provide a desired local effect to increase dental osteogenesis, mineralization or soft tissue regeneration in the oral cavity, because effective amounts for increasing dental osteogenesis, mineralization or soft tissue regeneration in the oral cavity with respect to GMSCs and PDLSCs were known for AA (see, e.g., Pham 2015, Yan 2013, Wei 2012, Yussif 2016 and Aytekin 2020), and that the combination with resveratrol was known to increase osteogenic differentiation of GMSCs and PDLSCs (Vidoni 2019 and Wang 2018). Moreover, in view of the overlapping pathways of action, the ordinary dentist would reasonably expect that resveratrol would increase proliferation of the stem cells by itself without vitamin C, particularly in view of Peltz 2012 teaching him that he could push the concentration of resveratrol up stepwise from 0.1 to 1, to 5 and then 10 μM, because “Resveratrol increased cell proliferation rate in a time and dosage dependent manner”, see Peltz 2012 at 4-5 and at 8, Figure 6.
The effective amount of each AA and resveratrol in his cocktail would represent a result effective variable that the ordinary dentist could tune through his ordinary and routine experimentation utilizing his ordinary skills at the time of filing to achieve the desired level of therapeutic effect for his patients. He might even determine that AA was not necessary, in view of the discussions in Chin 2017 that focused on resveratrol itself, and its therapeutic benefits for patients with periodontitis.
In either case, in view of the literature already supporting the use of both ascorbic acid and resveratrol for treating periodontitis, this dentist of just ordinary skill in the art at the time of filing would have a reasonable expectation of success that the combination would provide an additional benefit to his patients. See, e.g., the clear clinical success of the methods of Yussif 2016 with respect to inflammation, and at least Chin 2017 at 106 teaching that “Resveratrol suppresses P. gingivalis LPS–stimulated IκBα phosphorylation and nuclear translocation of the p65 subunit of NF-κB inhuman microvascular endothelial cells.”
This dentist of just ordinary skill in the art at the time of filing would also have a reasonable expectation of success in administering the combination in effective amounts in his patients’ oral cavities, because he could follow the clinical protocol laid out in Yussif 2016 for injecting the solution into the gingiva to access the GMSCs, or adapt it in such a way using his ordinary skills at the time of filing and just routine experimentation, so that the needle would penetrate deeper for the local injection to easily access the PDLSCs, as evident in the studies of Zhang 2018.
Accordingly, claims 1, 2, 3, 6, 7, and 10 were obvious at the time of filing.34
Regarding claims 4 and 5, resveratrol was known to modulate gene expression or protein expression of a component of the ERK/MAP kinase pathway (see, e.g., above figure from Chin 2017, or Kornicka 2017 at 954, or Vidoni 2019 at Fig. 4 for GMSCs (OCN, OPN clearly shown), or Wang 2018 at 7, Fig. 2 for PDLSCs (ALP, Runx2, Ocn clearly shown). Accordingly, the examiner assumes that resveratrol is an embodiment of these claims. They are rejected as obvious for the same reasons claims 1, 2, 3, 6, 7, and 10 were obvious at the time of filing.
Regarding claim 8, the dentist of just ordinary skill in the art at the time of filing would have a reasonable expectation of success in determining an effective amount of ascorbic acid to increase proliferation of the endogenous PDL stem cells, because such effective amounts were known in the art at the time of filing. See Wei 2012.
It would take only ordinary and routine experimentation for this ordinary dentist to determine if resveratrol would further stimulate proliferation of PDLSc by varying the effective amount already disclosed in Vidoni 2019 and Petlz 2012. Because PDLSCs were in the local vicinity of GMSCs, the ordinary dentist at the time of filing would reasonably be motivated to optimize the dose suited for GMSCs. The examiner notes that Vidoni 2019 already taught 5 μM was effective for GMSCs, and Peltz 2012 explicitly teaches 5 μM to increase proliferation.
Accordingly, claim 8 was obvious at the time of filing.
Regarding claim 9, patients with periodontitis were known to be at elevated risk of periodontal ligament or gingiva tissue damage, as the dentist of ordinary skill in the art at the time of filing would know from reading Chin 2017. See, e.g., Chin 2017 at 102, right column, discussion the disease progression of gingivitis to advanced periodontitis. The art had already established the method of treatment comprising local administration of a natural ERK activating compound as a stem cell therapy approach for treating periodontitis. See Zhang 2018. The dentist of just ordinary skill in the art at the time of filing would therefore have a reasonable expectation of success that, in following these procedures, the local administration of resveratrol with ascorbic acid would provide the therapeutic benefit to the same patient populations.
Accordingly, claim 9 was obvious at the time of filing.
Regarding claim 20, to the extent that it refers to selecting some sort of derivative of a natural ERK activating compound, Chin 2017 discloses “[t]herapuetic applications of resveratrol and its derivatives on periodontitis”. See Chin 2017 at Title. The ordinary dentist at the time of filing would have a reasonable expectation of success in using a derivative of resveratrol in his methods, such as the derivative disclosed in Chin 2017, designated “THSG”, because of its structural similarity to resveratrol, and because Chin 2017 at 106 teaches that “the efficiency of THSG in preventing induced periodontitis was better than that of resveratrol.”
Accordingly, claim 20 was obvious at the time of filing.
Secondary Considerations
Applicants only disclosed data pertain to in vitro studies. Applicant has not shown that any of these limited results extend to any therapeutic application. They are therefore not commensurate at all with the scope of these claims, that pertain to treating a subject in need.
Instead, Applicant chose to test obvious compounds, being resveratrol, curcumin, and PEITC. Not only were resveratrol and curcumin immediate choices for one seeking to treat periodontitis (see Chin 2017, indicating resveratrol was in clinical trials for periodontitis; see also Forouzanfar 2020, indicating curcumin already had known regenerative results for periodontitis), but also resveratrol and curcumin had been developed in a number of stem cell applications. See, e.g., Vidoni 2019, Wang 2018, and Sharifi 2020 for a discussion of resveratrol in stem cells; see also, a second article also by Sharifi – Sharifi, Simin, et al. "Stem cell therapy: Curcumin does the trick." Phytotherapy Research 33.11 (2019): 2927-2937, attached hereto. As it says in the title, for Stem cell therapy: “Curcumin does the trick”.
PEITC, as mentioned in the section regarding 112b, is a phenyl isothiocyanate just like Moringin, which was already well characterized in stem cells. PEITC itself was known to be effective against P. gingivalis, the bacteria primarily responsible for periodontitis. In view of the shared similarity between PEITC and Moringin, it was an obvious choice. See, e.g., Kim, Hyung Wook, et al. "Antibacterial activities of phenethyl isothiocyanate and its derivatives against human oral pathogens." Journal of the Korean Society for Applied Biological Chemistry 52.5 (2009): 555-559.
Had applicant actually presented preliminary data from an animal model, maybe there would have been unexpected results. But simply choosing obvious compounds known to be effective for treating periodontitis, that are also known to be effective in stem cell therapies, and then testing them in particular dental stem cells, represents simple obvious choices with a reasonable expectation of success.
Conclusion
No claims are allowed.
Any inquiry concerning this communication or earlier communications from the examiner should be directed to Christopher Evan Redwood whose telephone number is (571)272-8882. The examiner can normally be reached Monday - Friday 6:15 AM - 4:45 PM.
Examiner interviews are available via telephone, in-person, and video conferencing using a USPTO supplied web-based collaboration tool. To schedule an interview, applicant is encouraged to use the USPTO Automated Interview Request (AIR) at http://www.uspto.gov/interviewpractice.
If attempts to reach the examiner by telephone are unsuccessful, the examiner’s supervisor, Jeffrey S. Lundgren can be reached at 571-272-5541. The fax phone number for the organization where this application or proceeding is assigned is 571-273-8300.
Information regarding the status of published or unpublished applications may be obtained from Patent Center. Unpublished application information in Patent Center is available to registered users. To file and manage patent submissions in Patent Center, visit: https://patentcenter.uspto.gov. Visit https://www.uspto.gov/patents/apply/patent-center for more information about Patent Center and https://www.uspto.gov/patents/docx for information about filing in DOCX format. For additional questions, contact the Electronic Business Center (EBC) at 866-217-9197 (toll-free). If you would like assistance from a USPTO Customer Service Representative, call 800-786-9199 (IN USA OR CANADA) or 571-272-1000.
/C.E.R./ Examiner, Art Unit 1629
/JEFFREY S LUNDGREN/ Supervisory Patent Examiner, Art Unit 1629
1 Saud, Bhuvan, Rajani Malla, and Kanti Shrestha. "A review on the effect of plant extract on mesenchymal stem cell proliferation and differentiation." Stem cells international , vol. 2019, no. 1, article 7513404, pp. 1-13 (2019), hereinafter “Saud 2019”.
2 Sharifi, Simin, et al. "Phytochemicals impact on osteogenic differentiation of mesenchymal stem cells." BioFactors, vol. 46, no. 6, pp. 874-893 (Oct. 9, 2020), hereinafter “Sharifi 2020”.
3 Kornicka, Katarzyna, Ievgeniia Kocherova, and Krzysztof Marycz. "The effects of chosen plant extracts and compounds on mesenchymal stem cells—a bridge between molecular nutrition and regenerative medicine‐concise review." Phytotherapy Research, vol. 31, no. 7, pp. 947-958 (2017), hereinafter “Kornicka 2017”.
4 Xue, Wenqing, Jinhua Yu, and Wu Chen. "Plants and their bioactive constituents in mesenchymal stem cell‐based periodontal regeneration: A novel prospective." BioMed research international, vol. 2018, no. 1, article 7571363, pp. 1-15 (2018), hereinafter “Xue 2018”.
5 The “compounds” recited are resveratrol, “boswellic acid”, curcumin, and phenethyl isothiocyanate. The examiner notes that no data were provided for “boswellic acid”, nor any precise chemical name or structure for whatever it acid it refers to.
6 In the interests of compact prosecution, the examiner assumes these issues are typographical an Applicant means “natural ERK activating compound”.
7 Zhang, Xiuli, et al. "Local icariin application enhanced periodontal tissue regeneration and relieved local inflammation in a minipig model of periodontitis." International Journal of Oral Science, vol. 10, no. 2, article 19, pp. 1-6 (2018), hereinafter “Zhang 2018”.
8 Han, Nannan, et al. "Local application of IGFBP5 protein enhanced periodontal tissue regeneration via increasing the migration, cell proliferation and osteo/dentinogenic differentiation of mesenchymal stem cells in an inflammatory niche." Stem cell research & therapy, vol. 8, no. 1, article 210, pp. 1-13 (2017), hereinafter “Han 2017”.
9 Wang, Yuejun, et al. "IGFBP 5 enhances osteogenic differentiation potential of periodontal ligament stem cells and Wharton's jelly umbilical cord stem cells, via the JNK and MEK/Erk signalling pathways." Cell Proliferation, vol. 49, no. 5, pp. 618-627 (2016), hereinafter “Wang 2016”.
10 Forouzanfar, Fatemeh, et al. "Curcumin for the management of periodontal diseases: A review." Current pharmaceutical design, vol. 26, no. 34, pp. 4277-4284 (September 1, 2020), hereinafter “Forouzanfar 2020”.
11 Egusa, Hiroshi, et al. " Stem cells in dentistry – Part I: Stem cell sources." The Journal of Prosthodontic Research, vol. 56, no. 3, pp. 344-347 (2012), hereinafter “Egusa 2012”.
12 Yang, Ji Won, et al. "Therapeutic functions of stem cells from oral cavity: an update." International journal of molecular sciences, vol. 21, no. 12, article no. 4389, pp, 1-24 (2020), hereinafter “Yang 2020”.
13 Oz, Helieh S., and David A. Puleo. "Animal models for periodontal disease." BioMed Research International, vol. 2011, no. 1, article 754857, pp. 1-8 (2011), hereinafter “Oz 2011”.
14 MAPK pathway is explicitly discussed in Zhang 2018 at 4-5.
15 See Han 2017 at 3 (“Tumor necrosis factor alpha (TNFα) (Peprotech, Rocky Hill, NJ, USA) and rhIGFBP5 (R&D Systems, Minneapolis, MN, USA) were used to treat PDLSCs.”).
16 Yussif, Nermin M., Manar A. Abdul Aziz, and Ahmed R. Abdel Rahman. "Evaluation of the Anti‐Inflammatory Effect of Locally Delivered Vitamin C in the Treatment of Persistent Gingival Inflammation: Clinical and Histopathological Study." Journal of nutrition and metabolism, vol. 2016, no. 1, article 2978741, pp. 1-8 (2016), hereinafter “Yussif 2016”.
17 Yan, Yan, et al. "Vitamin C induces periodontal ligament progenitor cell differentiation via activation of ERK pathway mediated by PELP1." Protein & cell, vol. 4, no. 8, pp. 620-627 (2013), hereinafter “Yan 2013”.
18 Wei, Fulan, et al. "Vitamin C treatment promotes mesenchymal stem cell sheet formation and tissue regeneration by elevating telomerase activity." Journal of cellular physiology, vol. 227, no. 9, pp. 3216-3224 (2012), hereinafter “Wei 2012”.
19 Van Pham, Phuc, et al. "Vitamin C stimulates human gingival stem cell proliferation and expression of pluripotent markers." In Vitro Cellular & Developmental Biology-Animal, vol. 52, no. 2, pp. 218-227 (2016), hereinafter “Pham 2015”.
20 Fawzy El-Sayed, Karim M., Nhung Nguyen, and Christof E. Dörfer. "Ascorbic Acid, Inflammatory Cytokines (IL‐1β/TNF‐α/IFN‐γ), or Their Combination’s Effect on Stemness, Proliferation, and Differentiation of Gingival Mesenchymal Stem/Progenitor Cells." Stem Cells International, vol. 2020, no. 1, article 8897138, pp. 1-14 (Aug. 17, 2020), hereinafter “El-Sayed 2020”.
21 Aytekin, Zeliha, et al. "Immune modulatory and antioxidant effects of locally administrated vitamin C in experimental periodontitis in rats." Acta Odontologica Scandinavica, vol. 78, no. 6, pp. 425-432 (Aug. 17, 2020), hereinafter “Aytekin 2020”.
22 Previously cited in 112(b) section.
23 Peltz, Lindsay, et al. "Resveratrol exerts dosage and duration dependent effect on human mesenchymal stem cell development." PloS one, vol. 7, no. 5, article e37162 (2012), hereinafter “Peltz 2012”.
24 Wang, Yi-Jing, et al. "Resveratrol enhances the functionality and improves the regeneration of mesenchymal stem cell aggregates." Experimental & Molecular Medicine, vol. 50, no. 6, pp. 1-15 (2018), hereinafter “Wang 2018”.
25 Vidoni, Chiara, et al. "Autophagy drives osteogenic differentiation of human gingival mesenchymal stem cells." Cell Communication and Signaling, vol. 17, no. 1, article 98 (2019), pp. 1-17, hereinafter “Vidoni 2019”.
26 Chin, Yu‐Tang, et al. "Therapeutic applications of resveratrol and its derivatives on periodontitis." Annals of the New York Academy of Sciences, vol. 1403, no. 1, pp. 101-108 (2017), hereinafter “Chin 2017”.
27 Previously cited in 102/103 section.
28 Previously cited in 102/103 section.
29 Previously cited in 102/103 section.
30 Jia, Linglu, et al. "Comparative analysis of lncRNA and mRNA expression profiles between periodontal ligament stem cells and gingival mesenchymal stem cells." Gene, vol. 699, pp. 155-164 (2019), hereinafter “Linglu 2019”.
31 Previously cited in 102/103 section.
32 See Specification at 15-16, Paragraphs [00048]-[00049].
33 Peltz et al. is attached hereto, and is the Peltz 2012 reference.
34 The examiner notes that Applicant explicitly considers combinations with ascorbic acid as embodiments of the claims (see Specification at 7, item 1.21).
Prosecution Insights