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 .
1. Claims 1, 8, 10, 11, 13 – 15, 19 – 21, 24, 25, 28, 31, 33, 35, 36, 38, and 47 – 75 are pending.
Election/Restrictions
2. Applicant’s election without traverse of Group I (claims 1, 8, 10, 11, 13 – 15, 19 – 21, 24, 25, 28, 31, 33, 35, 36, and 47 – 75) in the reply filed on 12/15/2025 is acknowledged.
3. Claim 38 is withdrawn from further consideration pursuant to 37 CFR 1.142(b) as being drawn to a nonelected invention, there being no allowable generic or linking claim. Election was made without traverse in the reply filed on 12/15/2025.
Priority
4. This application is a national phase under 35 U.S.C. § 371 of International Application No. PCT/US2021/054462, filed October 12, 2021, which claims the benefit of priority to U.S. Provisional Patent Application Serial No. 63/090,590, filed October 12, 2020.
Information Disclosure Statement
5. The information disclosure statement (IDS) submitted on 09/17/2025 and 07/24/2023 are acknowledged. The submissions are in compliance with the provisions of 37 CFR 1.97. Accordingly, the information disclosure statements are being considered by the examiner.
6. The information disclosure statement filed 07/24/2023 fails to comply with 37 CFR 1.98(a)(2), which requires a legible copy of each cited foreign patent document; each non-patent literature publication or that portion which caused it to be listed; and all other information or that portion which caused it to be listed. It has been placed in the application file, but the information referred to therein has not been considered. C14 non-patent literature on page 4 of the IDS is missing.
Specification
7. The use of the term B-27 supplement, StemFlex, Matrigel, Neurobasal, Knockout serum replacement, Accutase and CellBanker, which is a trade name or a mark used in commerce, has been noted in this application. The term should be accompanied by the generic terminology; furthermore the term should be capitalized wherever it appears or, where appropriate, include a proper symbol indicating use in commerce such as ™, SM , or ® following the term.
Although the use of trade names and marks used in commerce (i.e., trademarks, service marks, certification marks, and collective marks) are permissible in patent applications, the proprietary nature of the marks should be respected and every effort made to prevent their use in any manner which might adversely affect their validity as commercial marks.
Claim Objections
8. Claim 1 is objected to because of the following informalities: in line 3, “the cells” should read “the stem or progenitor cells”. Appropriate correction is required.
9. Claim 19 is objected to because of the following informalities: in line 2, “the cells” should read “the stem or progenitor cells” to clarify that it is these cells that are contacted with the ROCK inhibitor and not the neural cells. Appropriate correction is required.
10. Claim 24 is objected to because of the following informalities: in line 1, “the cells” should read “the stem or progenitor cells” to clarify that it is these cells that are contacted with the ROCK inhibitor and not the neural cells. Appropriate correction is required.
11. Claim 24 is objected to because of the following informalities: in line 2, “the Compound” should read “the compound”. Appropriate correction is required.
12. Claim 33 is objected to because of the following informalities: in line 1, “a Compound” should read “the compound”. Appropriate correction is required.
13. Claim 67 is objected to because of the following informalities: in line 1, “the cells” should read “the neural cells” to clarify that it is the neural cells that express one or more neural cell markers and not the stem or progenitor cells. Appropriate correction is required.
14. Claim 73 is objected to because of the following informalities: in line 2, “PAX-6” should read “PAX6” because this is how it is first recited in claim 67. Appropriate correction is required.
15. Claim 74 is objected to because of the following informalities: in line 1, “wherein stem cells” should read “wherein the stem cells”. Appropriate correction is required.
Claim Rejections - 35 USC § 112
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.
16. Claims 24, 25, 33, and 63 – 66 are rejected under 35 U.S.C. 112(b) or 35 U.S.C. 112 (pre-AIA ), second paragraph, as being indefinite for failing to particularly point out and distinctly claim the subject matter which the inventor or a joint inventor (or for applications subject to pre-AIA 35 U.S.C. 112, the applicant), regards as the invention.
17. Claim 24 recites the limitation "the ROCK inhibitor" in line 2 – 3. There is insufficient antecedent basis for this limitation in the claim. Claim 25 is also rejected as it depends from claim 24 and does not clarify the grounds of rejection.
18. Claim 33 contains the trademark/trade name B-27. Where a trademark or trade name is used in a claim as a limitation to identify or describe a particular material or product, the claim does not comply with the requirements of 35 U.S.C. 112(b) or 35 U.S.C. 112 (pre-AIA ), second paragraph. See Ex parte Simpson, 218 USPQ 1020 (Bd. App. 1982). The claim scope is uncertain since the trademark or trade name cannot be used properly to identify any particular material or product. A trademark or trade name is used to identify a source of goods, and not the goods themselves. Thus, a trademark or trade name does not identify or describe the goods associated with the trademark or trade name. In the present case, the trademark/trade name is used to identify/describe a media supplement and, accordingly, the identification/description is indefinite.
19. Regarding claim 63, it is unclear how “the compound” is not a SMAD inhibitor when claim 1 requires selecting one of the recited compounds, all of which can be a SMAD inhibitor. For example, Hao (Hao, Jijun, et al. ACS chemical biology 5.2 (2010): 245-253.), which is cited on the IDS filed 07/23/2024 teaches DMH1 is a BMP4 inhibitor in Figure 4b and blocks Smad 1/5/8 phosphorylation but not Smad2 in Figure 4c and d. It is unclear if claim 63 is meant to further limit the required selection of one or more of DMH1, DMH2, K02288, or A8301.
20. Regarding claim 64, it is unclear how “the compound” is not a BMP4 inhibitor when claim 1 requires selecting one of the recited compounds, all of which can inhibit BMP4 signaling. For example, Hao (Hao, Jijun, et al. ACS chemical biology 5.2 (2010): 245-253.), which is cited on the IDS filed 07/23/2024 teaches DMH1 is a BMP4 inhibitor in Figure 4b and blocks Smad 1/5/8 phosphorylation but not Smad2 in Figure 4c and d. It is unclear if claim 64 is meant to further limit the required selection of one or more of DMH1, DMH2, K02288, or A8301.
21. Regarding claim 65, it is unclear how “the compound” can be neither a SMAD nor a BMP4 inhibitor when claim 1 requires selecting one of the recited compounds, all of which are either a BMP4 inhibitor or a SMAD inhibitor, or both. For example, Hao (Hao, Jijun, et al. ACS chemical biology 5.2 (2010): 245-253.), which is cited on the IDS filed 07/23/2024 teaches DMH1 is a BMP4 inhibitor in Figure 4b and blocks Smad 1/5/8 phosphorylation but not Smad2 in Figure 4c and d. Santivale (Sanvitale, Caroline E., et al. PloS one 8.4 (2013): e62721.), which is cited on the IDS filed 07/24/2023 teaches K02288 inhibits BMP signaling in Figure 4a as K02288 decreases BMP4 induced Smad 1/5/8 phosphorylation. Claim 1 requires selecting one or more of DMH1, DMH2, K02288, or A8301 as “the compound”. It is unclear if claim 65 is meant to further limit the required selection of one or more of DMH1, DMH2, K02288, or A8301.
22. Regarding claim 66, it is unclear if the claim requires “the compound” satisfy a, b, and c or only one of a, b, or c. It is unclear if claim 66 is meant to further limit the required selection of one or more of DMH1, DMH2, K02288, or A8301.
23. Regarding claim 74, it is unclear what is meant by “wherein stem cells are differentiated” because the claim does not require any active method steps. It is unclear if the claim is requiring further steps of contacting the stem cells with factors to differentiated the cells to neural cells. Claim 75 is also rejected as it depends from claim 74 and does not clarify the grounds of rejection.
Claim Interpretation
24. For the purpose of applying prior art, “a compound selected from DMH1, DMH2, K02288, A8301, or combinations thereof” of claim 1 is interpreted as a Markush group but the claim does not limit the method only to contacting the cells with one or more of the recited compounds as the claim recites “the method comprising”.
25. For the purpose of applying prior art, claims 63 – 65 are interpreted as the compound is DMH1, DMH2, K02288, or A8301 or combinations thereof because claim 1 requires selecting one or more of these recited compounds.
26. For the purpose of applying prior art, “wherein stem cells are differentiated” is interpreted as the stem cells of claim 1 are differentiated to neural cells.
Claim Rejections - 35 USC § 102
In the event the determination of the status of the application as subject to AIA 35 U.S.C. 102 and 103 (or as subject to pre-AIA 35 U.S.C. 102 and 103) is incorrect, any correction of the statutory basis (i.e., changing from AIA to pre-AIA ) for the rejection will not be considered a new ground of rejection if the prior art relied upon, and the rationale supporting the rejection, would be the same under either status.
The following is a quotation of the appropriate paragraphs of 35 U.S.C. 102 that form the basis for the rejections under this section made in this Office action:
A person shall be entitled to a patent unless –
(a)(1) the claimed invention was patented, described in a printed publication, or in public use, on sale, or otherwise available to the public before the effective filing date of the claimed invention.
(a)(2) the claimed invention was described in a patent issued under section 151, or in an application for patent published or deemed published under section 122(b), in which the patent or application, as the case may be, names another inventor and was effectively filed before the effective filing date of the claimed invention.
27. Claim(s) 1, 8, 13, 14, 19, 20, 21, 24, 31, 33, 47, 61 – 65, and 67 – 75 is/are rejected under 35 U.S.C. 102(a)(1) as being anticipated by Neely (Neely, M. Diana, et al. ACS chemical neuroscience 3.6 (2012): 482-491.), hereinafter Neely.
Claim 1 is drawn to a method for differentiating stem or progenitor cells into neural cells or for producing neural cells from stem or progenitor cells, the method comprising contacting the cells with a compound selected from DMH1, DMH2, K02288, A8301, or combinations thereof.
Regarding claims 1, 47, and 63 – 65, Neely teaches a method of producing neural cells comprising contacting stem cells with DMH1 (“DMH1”) (page 488, right col. para. 2 – 3).
Regarding claim 8, Neely teaches human iPSCs (hiPSCs) (page 488, right col. para. 2 – 3).
Regarding claim 13, Neely teaches in Figure 3 continuous contact with DMH1 for 7 days.
Regarding claim 14, Neely teaches 0.5 µM DMH1 (page 488, right col. para. 3).
Regarding claim 19, Neely teaches contacting the hiPSCs with a ROCK inhibitor (page 488, right col. para. 2).
Regarding claim 20, Neely teaches the ROCK inhibitor is Y-27632 (page 488, right col. para. 2).
Regarding claim 21, Neely teaches 10 µM Y-27632 (page 488, right col. para. 2).
Regarding claim 24, Neely teaches the hiPSCs are contacted with Y-27632 prior to contact with DMH1 (page 488, right col. para. 2 – 3).
Regarding claims 31, 61, and 62, Neely teaches the method produces dopaminergic neurons (page 488, right col. para. 3; Figure 6; page 486, right col. last para.).
Regarding claim 33, Neely teaches the culture medium comprises DMEM/F12, N2, and DMH1 (page 488, right col. para. 2 – 3).
Regarding claims 67 – 73, Neely teaches the cells express PAX6 (claim 67 and 73), OTX2 (claim 67, 68, and 72), SOX1 (claim 67, 69, and 73), tyrosine hydroxylase (claim 67 – 70), and tubulin (claims 67 – 71) (Figure 2C, 3, 4B, 5B, 6; page 485, right col.; page 486, right col. last para.).
Regarding claims 74 and 75, Neely teaches the hiPSCs (claim 75) are differentiated to dopaminergic neurons (claim 74) (page 488, right col. para. 2 – 3).
Therefore, Neely anticipates claims 1, 8, 13, 14, 19, 20, 21, 24, 31, 33, 47, 61 – 65, and 67 – 75.
28. Claim(s) 1, 8, 10, 13, 14, 28, 31, 33, 35, 36, 50, 63 – 71, 73, and 74 is/are rejected under 35 U.S.C. 102(a)(1) as being anticipated by Arshad (Valizadeh-Arshad, Zahra, et al. Cell Journal (Yakhteh) 19.4 (2017): 545.), hereinafter Arshad.
Claim 1 is drawn to a method for differentiating stem or progenitor cells into neural cells or for producing neural cells from stem or progenitor cells, the method comprising contacting the cells with a compound selected from DMH1, DMH2, K02288, A8301, or combinations thereof.
Regarding claims 1, 50, and 63 – 66, Arshad teaches a method of producing neural cells comprising contacting stem cells with A8301 (“A8301”) (page 546, left col. para. 3 – 4 and right col. para. 1).
Regarding claim 8 and 10, Arshad teaches ES cells (claim 8) that are human (hESCs) (claim 10) (page 546, left col. last para.).
Regarding claim 13, Arshad teaches hESCs were in continuous contact with A8301 for 4 days in Figure 1 (Stage 1).
Regarding claim 14, Arshad teaches 2 µM A8301 in Figure 1 (Stage 1).
Regarding claim 28, Arshad teaches the method excludes LDN193189 and SB431542 in Figure 1 (page 546, left col. para. 3 – 4 and right col. para. 1).
Regarding claim 31, Arshad teaches the method produces motor neurons (“motoneurons”) (page 547, right col. para. 4 – 5; Figure 1).
Regarding claim 33, Arshad teaches the culture medium comprises A8301 and DMEM/F12 (page 546, left col. last para.).
Regarding claim 35, Arshad teaches the neural cells are Nestin+, Pax6+, and Sox1+ in Figure 2F.
Regarding claim 36, Arshad teaches selecting the Nestin+, Pax6+, and Sox1+ neural cells for characterizing their electrophysiological properties (Figure 3; page 547, right col. last para.; page 548).
Regarding claims 67 – 71 and 73, Arshad teaches the cells express PAX6 (claim 67 and 73), SOX1 (claim 67, 68, and 73), MAP2 (claim 67, 69), TUJ1 (claim 67 – 70), and HB9 (claims 67 – 71) (Figure 2A – F; page 547, right col. para. 4).
Regarding claim 74, Arshad teaches the hESCs are differentiated to motor neurons (page 546, left col. para. 3 – 4 and right col. para. 1; Figure 1).
Therefore, Arshad anticipates claims 1, 8, 10, 13, 14, 28, 31, 33, 35, 36, 50, 63 – 71, 73, and 74.
29. Claim(s) 1, 8, 10, 13, 19, 20, 21, 24, 31, 49, 63 – 66, 67 – 71, 73, and 74 is/are rejected under 35 U.S.C. 102(a)(1) as being anticipated by Muguruma (Muguruma, Keiko, et al. Cell reports 10.4 (2015): 537-550.), hereinafter Muguruma.
Claim 1 is drawn to a method for differentiating stem or progenitor cells into neural cells or for producing neural cells from stem or progenitor cells, the method comprising contacting the cells with a compound selected from DMH1, DMH2, K02288, A8301, or combinations thereof.
Regarding claims 1, 49, and 63 – 65, Muguruma teaches a method of producing neural cells comprising contacting stem cells with K02288 (“K02288”) (page 542, right col. para. 1; Figure S3O and P; Figure 3E – I; page 549, right col. para. 4).
Regarding claim 8 and 10, Muguruma teaches ES cells (claim 8) that are human (claim 10) (page 542, right col. para. 1; page 549, right col. para. 4).
Regarding claim 13, Muguruma teaches in Figure S3O legend that cells were contacted continuously with K02288 for 4 days (Supplemental page 6).
Regarding claim 19, Muguruma teaches contacting the hESCs with a ROCK inhibitor (page 549, right col. para. 4).
Regarding claim 20, Muguruma teaches Y-27632 (page 549, right col. para. 4).
Regarding claim 21, Muguruma teaches 10 µM Y-27632 (page 549, right col. para. 4).
Regarding claim 24, Muguruma teaches the hESCs are contacted with Y27632 prior to contact with K02288 (page 549, right col. para. 4).
Regarding claim 31, Muguruma teaches the method produces granule cells and DCN neurons (“glutamatergic”) (Figure 3I; Figure S3; page 542, right col., para. 1).
Regarding claim 67 – 71 and 73, Muguruma teaches the neural cells express PAX6 (claims 67 and 73), ATOH1 (“neurofilament”) (claims 67 – 68), BRN2 (claims 67 – 69), MAP2 (claims 67 – 70, and TuJ (claims 67 – 71) (Figure 3B – H, R, S; Figure S3B).
Regarding claim 74, Muguruma teaches the hESCs are differentiated to granule cells and DCN neurons (Figure 3I; Figure S3; page 542, right col. para. 1).
Therefore, Muguruma anticipates claims 1, 8, 10, 13, 19, 20, 21, 24, 31, 49, 63 – 66, 67 – 71, 73, and 74 .
30. Claim(s) 1, 8, 10, 13, 28, 31, 33, 35, 36, 47, 61 – 65, and 67 – 74 is/are rejected under 35 U.S.C. 102(a)(1) as being anticipated by Gonzalez (US9926529-B2; Filed 04/23/2013; Published 03/27/2018), hereinafter Gonzalez which is cited on the IDS filed 07/24/2023.
Claim 1 is drawn to a method for differentiating stem or progenitor cells into neural cells or for producing neural cells from stem or progenitor cells, the method comprising contacting the cells with a compound selected from DMH1, DMH2, K02288, A8301, or combinations thereof.
Regarding claim 1, 47, and 63 – 65, Gonzalez teaches a method of producing neural cells from stem cells by contacting the stem cells with DMH1 (“DMH1”) (col. 17, lines 23 – 39; col. 18, lines 23 – 28).
Regarding claim 8 and 10, Gonzalez teaches iPSCs (claim 8) and human ES cells (claim 10) (col. 10 lines 5 – 8; col. 17, lines 55 – 56; col. 19, lines 27 – 31).
Regarding claim 13, Gonzalez teaches continuous contact of the hPSCs with DMH1 for 7 days (col. 17, lines 23 – 36; col. 18, lines 23 – 28).
Regarding claim 14, Gonzalez teaches 1 µM DMH1 (col. 18, lines 23 – 28).
Regarding claim 28, Gonzalez teaches the method excludes LDN193189 and SB431542 in Example 1 and 3 (col. 17, lines 23 – 36; col. 18, lines 23 – 28).
Regarding claim 31, 61, and 62, Gonzalez teaches dopaminergic neurons in Example 4 (col. 18, lines 40 – 67; col. 19., lines 1 – 26).
Regarding claim 33, Gonzalez teaches DMH1 in N2B27 medium comprising DMEM/F12 (col. 18, lines 23 – 28).
Regarding claim 35 and 36, Gonzalez teaches in Figure 3 that treatment of hPSCs with SB218078 and DMH1 yields cells that are Nestin+, Sox1+ and Pax6+ (claim 35) and that neural stem cell markers include nestin, PAX6, and SOX1 and that neural stem cells derived from hPSCs can be readily identified using methods well known to one of skill in the art including identifying neural stem cell markers using immunohistochemistry, FACS analysis, and measurement of RNA expression levels (claim 36) (col. 5, lines 7 – 9; col. 9, lines 38 – 52; col. 10, lines 9 – 13 and 63 – 67; col. 17, lines 33 – 39).
Regarding claims 67 – 73, Gonzalez teaches the cells are positive for Pax6 (claim 67 and 73), nestin (claims 67, 68, 73), BRN2 (claims 67 – 69), TUJ1 (claims 67 – 70), and FoxA2 (claims 67 – 72) (col. 17, lines 32 – 33; Figure 3 and 5B; col. 18, lines 52 – 64).
Regarding claim 74, Gonzalez teaches the stem cells are differentiated to dopamine neurons (col. 18, lines 40 – 63).
Therefore, Gonzalez anticipates claims 1, 8, 10, 13, 28, 31, 33, 35, 36, 47, 61 – 65, and 67 – 74 .
31. Claim(s) 1, 14, 28, 31, 33, 47, and 63 – 65 is/are rejected under 35 U.S.C. 102(a)(1) as being anticipated by Petersen (Petersen, Mark A., et al. Neuron 96.5 (2017): 1003-1012.), hereinafter Petersen.
Claim 1 is drawn to a method for differentiating stem or progenitor cells into neural cells or for producing neural cells from stem or progenitor cells, the method comprising contacting the cells with a compound selected from DMH1, DMH2, K02288, A8301, or combinations thereof.
Regarding claim 1, 47, and 63 – 65, Petersen teaches a method of treating oligodendrocyte progenitor cells (OPCs) with DMH1 (“DMH1”) to produce neural cells (page 1005, right col. para. 2).
Regarding claim 14, Petersen teaches 1 µM DMH1 (page 1017, para. 1).
Regarding claim 28, Petersen teaches the method excludes LDN193189 and SB431542 (page 1017, para. 1).
Regarding claim 31, Petersen teaches oligodendrocytes (page 1008, left col. para. 1).
Regarding claim 33, Petersen teaches DMH1 contacts OPCs in a culture medium comprising DMEM, B27, N2, and antibiotics (page 1016, para. 1; page 1017, para. 1).
Therefore, Petersen anticipates claims 1, 14, 28, 31, 33, 47, and 63 – 65.
Claim Rejections - 35 USC § 103
In the event the determination of the status of the application as subject to AIA 35 U.S.C. 102 and 103 (or as subject to pre-AIA 35 U.S.C. 102 and 103) is incorrect, any correction of the statutory basis (i.e., changing from AIA to pre-AIA ) for the rejection will not be considered a new ground of rejection if the prior art relied upon, and the rationale supporting the rejection, would be the same under either status.
The following is a quotation of 35 U.S.C. 103 which forms the basis for all obviousness rejections set forth in this Office action:
A patent for a claimed invention may not be obtained, notwithstanding that the claimed invention is not identically disclosed as set forth in section 102, if the differences between the claimed invention and the prior art are such that the claimed invention as a whole would have been obvious before the effective filing date of the claimed invention to a person having ordinary skill in the art to which the claimed invention pertains. Patentability shall not be negated by the manner in which the invention was made.
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.
32. Claim(s) 1, 8, 10, 11, 13, 14, 19, 20, 21, 24, 28, 31, 33, 35, 36, 47, 61 – 65 is/are rejected under 35 U.S.C. 103 as being unpatentable over Neely (Neely, M. Diana, et al. ACS chemical neuroscience 3.6 (2012): 482-491.), hereinafter Neely in view of Gonzalez (US9926529-B2; Filed 04/23/2013; Published 03/27/2018), hereinafter Gonzalez which is cited on the IDS filed 07/24/2023 in view of Rodin (Rodin, Sergey, et al Nature biotechnology 28.6 (2010): 611.), hereinafter Rodin.
Neely anticipates claims 1, 8, 13, 14, 19, 20, 21, 24, 31, 33, 47, 61 – 65, and 67 – 75 as set forth above. Neely does not teach differentiating human embryonic stem cells of claim 10 or the stem cells are HS420 cells of claim 11. Neely teaches the method excludes contacting stem cells with LDN193189 but teaches the method includes SB431542 that is required to be excluded in claim 28. Neely teaches the method produces cells that Sox1+ and Pax6+ (Figure 3; page 485, right col.) but does not teach Nestin+ cells of claims 35 and 36. However, Neely teaches the hiPSCs were derived from patient skin biopsies and validated (page 487, right col. last para.; page 488, left col.). Neely teaches in Figure 1 that measuring pluripotency of the hiPSCs lines against the human embryonic stem cell line HES-2 in Figure 1B where some hiPSC cell lines showed significantly reduced levels of the pluripotency markers (page 484, left col. last para.). Neely teaches recent studies have shown that the combined inhibition of the BMP and TGF-b1 pathways results in highly efficient conversion of human embryonic stem cells and hiPSCs into neural precursor cells (page 483, left col. para. 1). Neely teaches to ensure the successful use of hiPSC-derived neural precursors or differentiated neurons for cell-based therapy or drug discovery, standardized, chemically defined and economically affordable methods to derive these cells need to be developed (page 483, right col. last para.; page 484, left col. para. 1; page 487, right col. para. 1). Neely teaches hiPSCs allow for the possibility of studying human neurons derived from patients with neurological diseases and efficient and reproducible methods to differentiate hiPSCs into neurons are critical to ensure successful use of these cells for the study of human developmental processes, cellular mechanisms underlying human disease, or the development of drug screening platforms (Abstract; page 487, right col. para. 1).
Regarding claim 10 and 28, Gonzalez teaches a method of producing dopaminergic neurons from human embryonic stem cells (claim 10) by contacting the cells with SB218078 and DMH1 (claim 28) (col. 2, lines 44 – 47; col. 7, lines 22 – 38 and 45 – 53; col. 9, lines 10 – 15; col. 17, lines 23 – 39; col. 18, lines 23 – 28; col. 19, lines 27 – 28).
Regarding claim 35 and 36, Gonzalez teaches in Figure 3 that treatment of hPSCs with SB218078 and DMH1 yields cells that are Nestin+, Sox1+ and Pax6+ (claim 35) and that neural stem cell markers include nestin, PAX6, and SOX1 and that neural stem cells derived from hPSCs can be readily identified using methods well known to one of skill in the art including identifying neural stem cell markers using immunohistochemistry, FACS analysis, and measurement of RNA expression levels (claim 36) (col. 5, lines 7 – 9; col. 9, lines 38 – 52; col. 10, lines 9 – 13 and 63 – 67; col. 17, lines 33 – 39).
Gonzalez does not teach HS420 cells of claim 11. However, Gonzalez teaches a potential application of stem cells is making cells and tissues for medical therapies and stem cells offer the possibility of a renewable source of replacement cells and tissues to treat a myriad of diseases, conditions, and disabilities including Parkinson’s disease, which is a neurological disorder caused by a progressive degeneration of midbrain dopamine neurons (col. 1, lines 47 – 59). Gonzalez teaches there is currently no cure for Parkinson’s disease and the localized nature of the loss of dopamine neurons makes cell replacement therapy an attractive approach (col. 1, lines 62 – 67). Gonzalez teaches implantation of neuronal cells such as neural stem cells and dopamine neurons have already been shown to improve the motor symptoms in Parkinson’s disease animal models (col. 2, lines 1 – 4). Gonzalez teaches it is critical to be able to generate a homogenous population of NSCs which will in turn generate functional DAN neurons either in situ or in vitro, depending on whether the terminal differentiation occurs in the patient’s brain or in culture (col. 2, lines 5 – 10). Gonzalez teaches the method provides for the generation of neural stem cells from hPSCs that can be further differentiated into mature neurons for cell therapies or drug discovery (col. 6, lines 52 – 55). One would have been motivated to combine the teachings of Neely and Gonzalez because both teach methods of differentiating human pluripotent stem cells into neural cells for use in drug discovery targeting neurological diseases.
Regarding claim 11, Rodin teaches HS420 cells can be cultured for at least 4 months in a chemically defined medium and the cells self-renewed with normal karyotype and were capable of differentiation (Abstract; page 611, right col. last para.; page 612, left col. para. 1 and right col. para. 2 – 3; page 616, left col. para. 1). Rodin teaches there is a great need for chemically defined, xeno-free, feeder-free culture systems for hES cells (page 614, right col. para. 2).
It would have been obvious prior to the effective filing date of the invention as claimed for the person or ordinary skill in the art to combine the teachings of Neely regarding a method of producing dopaminergic neurons with DMH1 from hiPSCs with the teachings of Gonzalez regarding a method of producing dopamine neurons with DMH1 without LDN193189 or SB431542 from hES cells with the teachings of Rodin regarding HS420 hES cells can be cultured for a long period in a chemically defined media and maintain pluripotency to arrive at the claimed method where the cells are HS420 human embryonic stem cells and the method excludes LDN193189 and SB431542 and the neural cells that are Nestin+, Pax6+, and Sox1+ are selected to produce dopamine neurons. One would have been motivated to combine the teachings of Neely, Gonzalez, and Rodin in a method of producing dopamine neurons from a hES cell line instead of deriving pluripotent cells from skin biopsies for treating Parkinson’s disease as Gonzalez teaches a potential application of stem cells is making cells and tissues for medical therapies and stem cells offer the possibility of a renewable source of replacement cells and tissues to treat a myriad of diseases, conditions, and disabilities including Parkinson’s disease, which is a neurological disorder caused by a progressive degeneration of midbrain dopamine neurons and Gonzalez teaches there is currently no cure for Parkinson’s disease and the localized nature of the loss of dopamine neurons makes cell replacement therapy an attractive approach and Neely teaches the hiPSCs used in the method required derivation from skin biopsies and subsequent validation and Neely teaches standardized, chemically defined and economically affordable methods to derive neural precursors or differentiated neurons for cell-based therapy or drug discovery need to be developed and Rodin teaches there is a great need for chemically defined, xeno-free, feeder-free culture systems for hES cells. One would have a reasonable expectation of success in combining the teachings as Gonzalez teaches the method produces dopamine neurons from hES cells without LDN193189 or SB431542 and Rodin teaches HS420 cells can be cultured for at least 4 months in a chemically defined medium and the cells self-renewed with normal karyotype and were capable of differentiation.
33. Claim(s) 1, 8, 13, 14, 15, 19, 20, 21, 24, 31, 33, 47, 48, 51, 61 – 65 is/are rejected under 35 U.S.C. 103 as being unpatentable over Neely (Neely, M. Diana, et al. ACS chemical neuroscience 3.6 (2012): 482-491.), hereinafter Neely in view of Hao (Hao, Jijun, et al. ACS chemical biology 5.2 (2010): 245-253.), hereinafter Hao which is cited on the IDS filed 07/24/2023.
Neely anticipates claims 1, 8, 13, 14, 19, 20, 21, 24, 31, 33, 47, 61 – 65, and 67 – 75 as set forth above. Neely teaches the cells are contacted with 0.5 µM DMH1 and 10 µM SB431542 (page 488, right col. para. 3) but does not teach 0.2 µM of claim 15, the compound is DMH2 of claim 48 or the compound comprises DMH1 and DMH2 of claim 51. However, Neely teaches concomitant inhibition of the BMP and TGF-b1 branches of the TGF-b signaling pathways induces efficient neuralization of hiPSCs known as dual-SMAD inhibition (Abstract). Neely teaches DMH1 is a BMP-pathway inhibitor and SB431542 is a TGF-b1 inhibitor and treating hiPSCs with both compounds produces neural cells (Abstract; page 484, right col. last para; page 485, left col. para. 1). Neely teaches to ensure the successful use of hiPSC-derived neural precursors or differentiated neurons for cell-based therapy or drug discovery, standardized, chemically defined and economically affordable methods to derive these cells need to be developed (page 483, right col. last para.; page 484, left col. para. 1; page 487, right col. para. 1). Neely teaches hiPSCs allow for the possibility of studying human neurons derived from patients with neurological diseases and efficient and reproducible methods to differentiate hiPSCs into neurons are critical to ensure successful use of these cells for the study of human developmental processes, cellular mechanisms underlying human disease, or the development of drug screening platforms (Abstract; page 487, right col. para. 1).
Regarding claim 15, Hao teaches in Figure 4b DMH1 inhibited BMP signaling in a dose-dependent manner with inhibition at 0.2 µM only slightly less than at 0.5 µM. Hao teaches the IC50 for inhibition of BMP signaling by DMH1 is approximately 100 nM (page 4, para. 4).
Regarding claim 48, Hao teaches in Table 2 that DMH2 is both a BMP signaling inhibitor and a TGF-b1 signaling inhibitor. Hao teaches the IC50 for inhibition of BMP signaling by DMH1 is approximately 100 nM whereas the IC50 for DMH2 is 20 nM (page 4, para. 4).
Regarding claim 51, Hao teaches in Table 2 that DMH1 is a BMP signaling inhibitor but not a TGF-b1 signaling inhibitor and DMH2 is an inhibitor of TGF-b1 signaling where the IC50 for TGF-b1 inhibition by DMH2 is 1578 nM.
It would have been obvious prior to the effective filing date of the invention as claimed for the person of ordinary skill in the art to combine the teachings of Neely regarding a method of producing dopaminergic neurons with 0.5 µM DMH1 from hiPSCs where DMH1 is a BMP pathway inhibitor with the teachings of Hao regarding the IC50 for inhibition of BMP signaling by DMH1 is approximately 100 nM and DMH1 inhibited BMP signaling at 0.2 µM, which is only slightly less inhibition than at 0.5 µM DMH1 to arrive at the claimed method where the cells are contacted with 0.2 µM DMH1. One would have been motivated to combine the teachings of Neely and Hao in a cost-effective method of producing dopamine neurons for drug discovery where the concentration of DMH1 is lowered from 0.5 µM DMH1 to 0.2 µM DMH1as Neely teaches to ensure the successful use of hiPSC-derived neural precursors or differentiated neurons for cell-based therapy or drug discovery, standardized, chemically defined and economically affordable methods to derive these cells need to be developed. One would have a reasonable expectation of success in combining the teachings as Hao teaches DMH1 inhibited BMP signaling in a dose-dependent manner with inhibition at 0.2 µM only slightly less than at 0.5 µM and Hao teaches the IC50 for inhibition of BMP signaling by DMH1 is approximately 100 nM.
It would have been obvious prior to the effective filing date of the invention as claimed for the person of ordinary skill in the art to combine the teachings of Neely regarding a method of producing dopaminergic neurons using the BMP inhibitor DMH1 and the TGF-b1 inhibitor SB431542 with the teachings of Hao regarding DMH2 is both a BMP signaling inhibitor and a TGF-b1 signaling inhibitor and the IC50 for inhibition of BMP signaling by DMH1 is approximately 100 nM whereas the IC50 for DMH2 is 20 nM to arrive at the claimed method where DMH1 in the method of Neely is substituted for DMH2 of Hao. One would have been motivated to make the substitution for a cost-effective method of producing dopamine neurons for drug discovery because Neely teaches to ensure the successful use of hiPSC-derived neural precursors or differentiated neurons for cell-based therapy or drug discovery, standardized, chemically defined and economically affordable methods to derive these cells need to be developed and Hao teaches the IC50 for BMP inhibition by DMH2 is lower than that of DMH1. One would have a reasonable expectation of success in combining the teachings as Hao teaches DMH2 is a BMP signaling inhibitor and has a lower IC50 for BMP signaling inhibition.
It would have been obvious prior to the effective filing date of the invention as claimed for the person of ordinary skill in the art to combine the teachings of Neely regarding a method of producing dopaminergic neurons using the BMP inhibitor DMH1 and the 10 µM TGF-b1 inhibitor SB431542 with the teachings of Hao regarding DMH1 is a BMP signaling inhibitor and DMH2 is a TGF-b1 signaling inhibitor to arrive at the claimed method wherein the compound comprises DMH1 and DMH2 where SB431542 in the method of Neely is substituted for DMH2. One would have been motivated to make the substitution for a cost-effective method of producing dopamine neurons for drug discovery because Neely teaches to ensure the successful use of hiPSC-derived neural precursors or differentiated neurons for cell-based therapy or drug discovery, standardized, chemically defined and economically affordable methods to derive these cells need to be developed. One would have a reasonable expectation of success in combining the teachings as Hao teaches the IC50 for TGF-b1 signaling by DMH2 is 1578 nM and thus a DMH2 concentration lower than 10 µM can be used in the method.
34. Claim(s) 1, 8, 13, 14, 19, 20, 21, 24, 25, 31, 33, 47, 61 – 65, and 67 – 75 is/are rejected under 35 U.S.C. 103 as being unpatentable over Neely (Neely, M. Diana, et al. ACS chemical neuroscience 3.6 (2012): 482-491.), hereinafter Neely in view of Cosset (Cosset, Érika, et al. Biomaterials 53 (2015): 296-308.), hereinafter Cosset.
Neely anticipates claims 1, 8, 13, 14, 19, 20, 21, 24, 31, 33, 47, 61 – 65, and 67 – 75 as set forth above. Neely teaches treating the cells with a ROCK inhibitor for 4 – 6 days on Matrigel to allow the cells to reach confluence prior to contacting the cells with DMH1 (page 488, right col. para. 2) but does not teach the cells are contacted with the ROCK inhibitor for 1 – 48 h of claim 25. However, Neely teaches concomitant inhibition of the BMP and TGF-b1 branches of the TGF-b signaling pathways induces efficient neuralization of hiPSCs known as dual-SMAD inhibition (Abstract). Neely teaches DMH1 is a BMP-pathway inhibitor and SB431542 is a TGF-b1 inhibitor and treating hiPSCs with both compounds produces neural cells (Abstract; page 484, right col. last para; page 485, left col. para. 1). Neely teaches DMH1 is an analogue of dorsomorphin (page 484, left col. para. 1). Neely teaches to ensure the successful use of hiPSC-derived neural precursors or differentiated neurons for cell-based therapy or drug discovery, standardized, chemically defined and economically affordable methods to derive these cells need to be developed (page 483, right col. last para.; page 484, left col. para. 1; page 487, right col. para. 1). Neely teaches hiPSCs allow for the possibility of studying human neurons derived from patients with neurological diseases and efficient and reproducible methods to differentiate hiPSCs into neurons are critical to ensure successful use of these cells for the study of human developmental processes, cellular mechanisms underlying human disease, or the development of drug screening platforms (Abstract; page 487, right col. para. 1).
Cosset teaches a method of producing neural cells from hES cells (hESCs) comprising treating the hESCs with 10 µM ROCK inhibitor Y27632 for one day to reduce hESC cell death followed by treating the cells with the dual-SMAD SB431542 and Dorsomorphin for 7 days producing cell aggregates (page 299, right col. para. 2).
It would have been obvious prior to the effective filing date of the invention as claimed for the person of ordinary skill in the art to combine the teachings of Neely regarding a method of a method of producing dopaminergic neurons using the BMP inhibitor DMH1 and the TGF-b1 inhibitor SB431542 where cells were treated with ROCK inhibitor for 4 – 6 days prior to DMH1 and SB431542 with the teachings of Cosset regarding a method of generating neural cells by treating stem cells with ROCK inhibitor for one day prior to treating with dual-SMAD inhibitors SB431542 and Dorsomorphin to arrive at the claimed method where the stem cells are contacted with the ROCK inhibitor for a time period of 24 hours before contacting the cells with DMH1 and SB431542. One would have been motivated to combine the teachings of Neely and Cosset in a cost-effective method of producing neural cells for drug discovery that shortens the method time by 3 – 5 days as Neely teaches to ensure the successful use of hiPSC-derived neural precursors or differentiated neurons for cell-based therapy or drug discovery, standardized, chemically defined and economically affordable methods to derive these cells need to be developed. One would have a reasonable expectation of success in combining the teachings as the method of Neely and the method of Cosset use SB431542 and Neely teaches DMH1 is an analogue of Dorsomorphin used in the method of Cosset.
35. Claim(s) 1, 14, 28, 31, 33, 47, 54, and 63 – 65 is/are rejected under 35 U.S.C. 103 as being unpatentable over Petersen (Petersen, Mark A., et al. Neuron 96.5 (2017): 1003-1012.), hereinafter Petersen in view of Muguruma (Muguruma, Keiko, et al. Cell reports 10.4 (2015): 537-550.), hereinafter Muguruma in view of Albert (Albert, Monika, et al. Brain Pathology 27.6 (2017): 737-747.), hereinafter Albert as evidenced by de Leon (de Leon AS, Das JM. Neuroanatomy, Dentate Nucleus. [Updated 2023 Jul 24]. In: StatPearls [Internet]. Treasure Island (FL): StatPearls Publishing; 2025 Jan-. Available from: https://www.ncbi.nlm.nih.gov/books/NBK554381/), hereinafter de Leon.
Petersen anticipates claims 1, 14, 28, 31, 33, 47, and 63 – 65 as set forth above. Petersen does not teach the compound comprises DMH1 and K02288 of claim 54. However, Petersen teaches fibrinogen activates BMP signaling pathway in oligodendrocyte progenitor cells (OPCs) and suppresses remyelination (Summary; page 1008, left col. last para. and right col. para. 2; Figure 4E). Petersen teaches fibrinogen induces phosphorylation of Smad 1/5/8 and inhibits OPC differentiation into myelinating oligodendrocytes and the effects of fibrinogen are rescued by BMP type I receptor inhibition using DMH1 (Summary; page 1005, left col. para. 3; page 1005, right col. para. 2; Figure 2D, E). Petersen teaches the BMP target Id2 is increased in demyelinated multiple sclerosis (MS) lesions and therapeutic depletion of fibrinogen decreases BMP signaling and enhances remyelination in vivo (Summary). Petersen teaches targeting fibrinogen may be an upstream therapeutic strategy to promote the regenerative potential of CNS progenitors in diseases with remyelination failure (Summary). Petersen teaches remyelination is critical for recovery in several diseases such as MS and OPCs often fail to differentiate into mature oligodendrocytes required for myelin repair (page 1004, right col.). Petersen teaches fibrinogen is deposited in MS, stroke, traumatic brain injury, and Alzheimer’s disease and fibrinogen is found in active and chronic MS lesions (page 1005, left col. para. 2). Petersen teaches fibrinogen is a driver of neuropathology promoting neuroinflammation and glial scar formation by direct effects on microglia, astrocytes, and neurons (page 1005, left col. para. 2). Petersen teaches fibrinogen increased expression of several BMP-responsive genes including Id1, Id2, Nog, Hes1, Hey1, and Lef1 that are associated with impaired OPC differentiation and upregulated in some MS patients (page 1005, right col. para. 1; Figure 1F). Petersen teaches active lesions of MS patients had high levels of Id2 in areas of fibrinogen deposition and demyelination (page 1005, right col. para. 1). Petersen teaches DMH1 is a dorsomorphin analog that inhibits ACVR1 (Alk2) and it blocked fibrinogen-induced phosphorylation of Smad1/5 and suppressed the Id genes in cultures of OPCs (page 1005, right col. para. 2; Figure 2D, E). Petersen teaches DMH1 increased the number of mature oligodendrocytes (page 1008, left col. para. 1). Petersen teaches depletion of fibrinogen in vivo reduced phospho-Smad 1/5/8 in demyelinated lesions suggesting reduction of BMP receptor pathway activation concomitant with increased mature oligodendrocytes and enhanced remyelination (page 1008, left col. para. 2).
Regarding “K02288”, Muguruma teaches a method of producing granule cells and deep cerebellar neurons (DCN) comprising contacting stem cells with K02288 (Figure 3E – I; Figure S3; page 542, right col. para. 1; Figure S3O and P; Figure 3I; page 544, left col. para. 1 – 2; page 549, right col. para. 4). Muguruma teaches K0228 is a BMP inhibitor (page S6).
Albert teaches the dentate nucleus is the predilection site for lesion development in MS (Abstract; page 737, left col. para. 1). Albert teaches a significant reduction of synapses in the dentate nucleus of MS patients with or without the presence of demyelination (Abstract; page 741; Figure 1; page 742, left col. para. 1 and last para.; page 746, left col. para. 3). Albert teaches neuronal loss in the dentate nucleus was more pronounced in cases with local demyelination (page 743, right col.). The dentate nucleus is the largest deep cerebellar cluster of neurons as evidenced by de Leon (page 1, para. 1 – 2).
It would have been obvious prior to the effective filing date of the invention as claimed for the person of ordinary skill in the art to combine the teachings of Petersen regarding a method of producing oligodendrocytes from OPCs by rescuing the effects of fibrinogen with DMH1 with the teachings of Muguruma regarding a method of producing deep cerebellar neurons by contacting stem cells with K02288 with the teachings of Albert regarding MS patients have reduced synapses in the dentate nucleus to arrive at the claimed method where the compound comprises DMH1 and K02288. One would have been motivated to combine the teachings of Petersen, Muguruma, and Albert in a method to treat multiple sclerosis with oligodendrocytes and deep cerebellar neurons as Petersen teaches targeting fibrinogen may be an upstream therapeutic strategy to promote the regenerative potential of CNS progenitors in diseases with remyelination failure and Petersen teaches remyelination is critical for recovery in several diseases such as MS and OPCs often fail to differentiate into mature oligodendrocytes required for myelin repair and Petersen teaches fibrinogen is deposited in MS, found in active and chronic MS lesions, and is a driver of neuropathology promoting neuroinflammation and glial scar formation by direct effects on microglia, astrocytes, and neurons and Albert teaches neuronal loss in the dentate nucleus was more pronounced in cases with local demyelination. One would have a reasonable expectation of success in combining the teachings as Petersen teaches decreasing BMP signaling enhances remyelination in vivo and DMH1 blocks the effects of fibrinogen and Muguruma teaches K02288 treatment produces deep cerebellar neurons and Muguruma teaches K02288 is a BMP inhibitor.
36. Claim(s) 1, 14, 28, 31, 33, 47, 55, and 63 – 65 is/are rejected under 35 U.S.C. 103 as being unpatentable over Petersen (Petersen, Mark A., et al. Neuron 96.5 (2017): 1003-1012.), hereinafter Petersen in view of Arshad (Valizadeh-Arshad, Zahra, et al. Cell Journal (Yakhteh) 19.4 (2017): 545.), hereinafter Arshad in view of Vogt (Vogt J, et. al. Ann Neurol. 2009 Sep;66(3):310-22.), hereinafter Vogt.
Petersen anticipates claims 1, 14, 28, 31, 33, 47, and 63 – 65 as set forth above. Petersen does not teach the compound comprises DMH1 and A8301 of claim 55. However, Petersen teaches fibrinogen activates BMP signaling pathway in oligodendrocyte progenitor cells (OPCs) and suppresses remyelination (Summary; page 1008, left col. last para. and right col. para. 2; Figure 4E). Petersen teaches fibrinogen induces phosphorylation of Smad 1/5/8 and inhibits OPC differentiation into myelinating oligodendrocytes and the effects of fibrinogen are rescued by BMP type I receptor inhibition using DMH1 (Summary; page 1005, left col. para. 3; page 1005, right col. para. 2; Figure 2D, E). Petersen teaches the BMP target Id2 is increased in demyelinated multiple sclerosis (MS) lesions and therapeutic depletion of fibrinogen decreases BMP signaling and enhances remyelination in vivo (Summary). Petersen teaches targeting fibrinogen may be an upstream therapeutic strategy to promote the regenerative potential of CNS progenitors in diseases with remyelination failure (Summary). Petersen teaches remyelination is critical for recovery in several diseases such as MS and OPCs often fail to differentiate into mature oligodendrocytes required for myelin repair (page 1004, right col.). Petersen teaches fibrinogen is deposited in MS, stroke, traumatic brain injury, and Alzheimer’s disease and fibrinogen is found in active and chronic MS lesions (page 1005, left col. para. 2). Petersen teaches fibrinogen is a driver of neuropathology promoting neuroinflammation and glial scar formation by direct effects on microglia, astrocytes, and neurons (page 1005, left col. para. 2). Petersen teaches fibrinogen increased expression of several BMP-responsive genes including Id1, Id2, Nog, Hes1, Hey1, and Lef1 that are associated with impaired OPC differentiation and upregulated in some MS patients (page 1005, right col. para. 1; Figure 1F). Petersen teaches active lesions of MS patients had high levels of Id2 in areas of fibrinogen deposition and demyelination (page 1005, right col. para. 1). Petersen teaches DMH1 is a dorsomorphin analog that inhibits ACVR1 (Alk2) and it blocked fibrinogen-induced phosphorylation of Smad1/5 and suppressed the Id genes in cultures of OPCs (page 1005, right col. para. 2; Figure 2D, E). Petersen teaches DMH1 increased the number of mature oligodendrocytes (page 1008, left col. para. 1). Petersen teaches depletion of fibrinogen in vivo reduced phospho-Smad 1/5/8 in demyelinated lesions suggesting reduction of BMP receptor pathway activation concomitant with increased mature oligodendrocytes and enhanced remyelination (page 1008, left col. para. 2).
Regarding “A8301”, Arshad teaches a method of producing motor neurons comprising contacting stem cells with A8301 (page 546, left col. para. 3 – 4 and right col. para. 1; page 547, right col. para. 4 – 5; Figure 1). Arshad teaches motor neuron differentiation from hESCs is a goal of regenerative medicine to provide cell therapy as treatments for diseases that damage motor neurons (Abstract).
Vogt teaches MS patients have a massive loss of lower motor neurons (Abstract; page 312, right col. para. 2; Figure 1E, F). Vogt teaches spinal motor neurons in chronic active MS exhibited morphological signs of apoptosis (page 312, right col. para. 2). Vogt teaches the assumption that MS is a purely demyelinating disease has been challenged by the observation that axonal damage is a common finding in the CNS of MS patients (page 310, left col. and right col. para. 1). Vogt teaches because the cell death of lower motor neurons plays a prominent role in MS pathology, there is a reason to pursue scientific efforts to develop novel therapeutics strategies with neuroprotective potential (page 321, left col. para. 3).
It would have been obvious prior to the effective filing date of the invention as claimed for the person of ordinary skill in the art to combine the teachings of Petersen regarding a method of producing oligodendrocytes from OPCs by rescuing the effects of fibrinogen with DMH1 with the teachings of Arshad regarding a method of producing motor neurons by contacting stem cells with A8301 with the teachings of Vogt regarding MS patients have fewer motor neurons to arrive at the claimed method where the compound comprises DMH1 and A8301. One would have been motivated to combine the teachings of Petersen, Arshad, and Vogt in a method to treat multiple sclerosis with oligodendrocytes and motor neruons as Petersen teaches targeting fibrinogen may be an upstream therapeutic strategy to promote the regenerative potential of CNS progenitors in diseases with remyelination failure and Petersen teaches remyelination is critical for recovery in several diseases such as MS and OPCs often fail to differentiate into mature oligodendrocytes required for myelin repair and Petersen teaches fibrinogen is deposited in MS, found in active and chronic MS lesions, and is a driver of neuropathology promoting neuroinflammation and glial scar formation by direct effects on microglia, astrocytes, and neurons and Vogt teaches because the cell death of lower motor neurons plays a prominent role in MS pathology, there is a reason to pursue scientific efforts to develop novel therapeutics strategies with neuroprotective potential. One would have a reasonable expectation of success in combining the teachings as Petersen teaches decreasing BMP signaling enhances remyelination in vivo and DMH1 blocks the effects of fibrinogen and Arshad teaches A8301 treatment produces motor neurons.
37. Claim(s) 1, 8, 10, 13, 19, 20, 21, 24, 31, 49, 56, 63 – 66, 67 – 71, 73, is/are rejected under 35 U.S.C. 103 as being unpatentable over Muguruma (Muguruma, Keiko, et al. Cell reports 10.4 (2015): 537-550.), hereinafter Muguruma in view of Albert (Albert, Monika, et al. Brain Pathology 27.6 (2017): 737-747.), hereinafter Albert as evidenced by de Leon (de Leon AS, Das JM. Neuroanatomy, Dentate Nucleus. [Updated 2023 Jul 24]. In: StatPearls [Internet]. Treasure Island (FL): StatPearls Publishing; 2025 Jan-. Available from: https://www.ncbi.nlm.nih.gov/books/NBK554381/), hereinafter de Leon in view of Arshad (Valizadeh-Arshad, Zahra, et al. Cell Journal (Yakhteh) 19.4 (2017): 545.), hereinafter Arshad in view of Vogt (Vogt J, et. al. Ann Neurol. 2009 Sep;66(3):310-22.), hereinafter Vogt.
Muguruma anticipates claims 1, 8, 10, 13, 19, 20, 21, 24, 31, 49, 63 – 66, 67 – 71, 73, and 74 as set forth above. Muguruma does not teach the compound comprises K02288 and A8301 of claim 56. However, Muguruma teaches a method of producing granule cells and deep cerebellar neurons (DCN) comprising contacting stem cells with K02288 (Figure 3E – I; Figure S3; page 542, right col. para. 1; Figure S3O and P; Figure 3I; page 544, left col. para. 1 – 2; page 549, right col. para. 4). Muguruma teaches K0228 is a BMP inhibitor (page S6).
Albert teaches the dentate nucleus is the predilection site for lesion development in multiple sclerosis (MS) (Abstract; page 737, left col. para. 1). Albert teaches a significant reduction of synapses in the dentate nucleus of MS patients (Abstract; page 741; Figure 1; page 742, left col. para. 1 and last para.; page 746, left col. para. 3). Albert teaches neuronal loss in the dentate nucleus was more pronounced in cases with local demyelination (page 743, right col.). The dentate nucleus is the largest deep cerebellar cluster of neurons as evidenced by de Leon (page 1, para. 1 – 2). Albert does not teach “A8301”. One would have been motivated to combine the teachings of Muguruma and Albert in a method of treating MS because Muguuruma teaches a method of producing deep cerebellar neurons with K02288 and Albert teaches MS patients have neuronal loss in the dentate nucleus.
Regarding “A8301”, Arshad teaches a method of producing motor neurons comprising contacting stem cells with A8301 (page 546, left col. para. 3 – 4 and right col. para. 1; page 547, right col. para. 4 – 5; Figure 1). Arshad teaches motor neuron differentiation from hESCs is a goal of regenerative medicine to provide cell therapy as treatments for diseases that damage motor neurons (Abstract).
Vogt teaches MS patients have a massive loss of lower motor neurons (Abstract; page 312, right col. para. 2; Figure 1E, F). Vogt teaches spinal motor neurons in chronic active MS exhibited morphological signs of apoptosis (page 312, right col. para. 2). Vogt teaches the assumption that MS is a purely demyelinating disease has been challenged by the observation that axonal damage is a common finding in the CNS of MS patients (page 310, left col. and right col. para. 1). Vogt teaches because the cell death of lower motor neurons plays a prominent role in MS pathology, there is a reason to pursue scientific efforts to develop novel therapeutics strategies with neuroprotective potential (page 321, left col. para. 3).
It would have been obvious prior to the effective filing date of the invention as claimed for the person of ordinary skill in the art to combine the teachings of Muguruma regarding a method of producing deep cerebellar neurons by contacting stem cells with K02288 with the teachings of Albert regarding MS patients have reduced synapses in the dentate nucleus Arshad regarding a method of producing motor neurons by contacting stem cells with A8301 with the teachings of Vogt regarding MS patients have fewer motor neurons to arrive at the claimed method where the compound comprises K02288 and A8301. One would have been motivated to combine the teachings of Muguruma, Albert, Arshad, and Vogt in a method to treat multiple sclerosis with deep cerebellar neurons and motor neurons as Muguuruma teaches a method of producing deep cerebellar neurons with K02288 and Albert teaches MS patients have neuronal loss in the dentate nucleus and Arshad teaches motor neuron differentiation from hESCs is a goal of regenerative medicine to provide cell therapy as treatments for diseases that damage motor neurons and Albert teaches MS patients have neuronal loss. One would have a reasonable expectation of success in combining the teachings as Muguruma teaches K02288 treatment produces deep cerebellar neurons and Arshad teaches the method produces motor neurons.
38. Claim(s) 1, 8, 13, 14, 19, 20, 21, 24, 31, 33, 47, 52, 57, 61 – 65, and 67 – 75 is/are rejected under 35 U.S.C. 103 as being unpatentable over Neely (Neely, M. Diana, et al. ACS chemical neuroscience 3.6 (2012): 482-491.), hereinafter Neely in view of Hao (Hao, Jijun, et al. ACS chemical biology 5.2 (2010): 245-253.), hereinafter Hao which is cited on the IDS filed 07/24/2023 in view of Kriks (Kriks, Sonja, et al. Nature 480.7378 (2011): 547-551.), hereinafter Kriks in view of Muguruma (Muguruma, Keiko, et al. Cell reports 10.4 (2015): 537-550.), hereinafter Muguruma in view of He (He N, et. al. NMR Biomed. 2017 Apr;30(4):10.1002/nbm.3554), hereinafter He as evidenced by de Leon (de Leon AS, Das JM. Neuroanatomy, Dentate Nucleus. [Updated 2023 Jul 24]. In: StatPearls [Internet]. Treasure Island (FL): StatPearls Publishing; 2025 Jan-. Available from: https://www.ncbi.nlm.nih.gov/books/NBK554381/), hereinafter de Leon.
Neely anticipates claims 1, 8, 13, 14, 19, 20, 21, 24, 31, 33, 47, 61 – 65, and 67 – 75 as set forth above. Neely does not teach the compound comprises K02288 and DMH2 of claim 52 or the compound comprises DMH1, DMH2, and K0228 of claim 57. However, Neely teaches concomitant inhibition of the BMP and TGF-b1 branches of the TGF-b signaling pathways induces efficient neuralization of hiPSCs known as dual-SMAD inhibition (Abstract). Neely teaches DMH1 is a BMP-pathway inhibitor and SB431542 is a TGF-b1 inhibitor and treating hiPSCs with both compounds produces neural cells (Abstract; page 484, right col. last para; page 485, left col. para. 1). Neely teaches the method produces dopaminergic neurons where the neural induction media comprised DMH1 but not SB431542 (page 488, right col. para. 2 – 3; Figure 6; page 486, right col. last para.). Neely teaches the neuralization medium further included SHH, BDNF, FGF8b, GDNF, TGFb-3, ascorbate, and dcAMP (page 488, right col. para. 3). Neely teaches to ensure the successful use of hiPSC-derived neural precursors or differentiated neurons for cell-based therapy or drug discovery, standardized, chemically defined and economically affordable methods to derive these cells need to be developed (page 483, right col. last para.; page 484, left col. para. 1; page 487, right col. para. 1). Neely teaches hiPSCs allow for the possibility of studying human neurons derived from patients with neurological diseases and efficient and reproducible methods to differentiate hiPSCs into neurons are critical to ensure successful use of these cells for the study of human developmental processes, cellular mechanisms underlying human disease, or the development of drug screening platforms (Abstract; page 487, right col. para. 1).
Regarding “DMH2” of claims 52 and 57, Hao teaches in Table 2 that DMH1 is a BMP signaling inhibitor but not a TGF-b1 signaling inhibitor and DMH2 is an inhibitor of BMP and TGF-b1 signaling where the IC50 for TGF-b1 inhibition by DMH2 is 1578 nM. Hao teaches the IC50 for BMP inhibition (ALK2) by DMH2 (42.8 nM) is similar to that for LDN193189 (40.7 nM) in Table 2. Hao does not teach “K02288”. One would have been motivated to combine the teachings of Neely and Hao to substitute 10 µM SB431542 in the method of Neely with a lower concentration of DMH2 to make the method of producing dopamine neurons more economically affordable.
Kriks teaches a method of deriving dopamine neurons from hESCs or hPSCs comprising dual-SMAD inhibition to accelerate neural fate acquisition with LDN193189 and SB43152 (10 µM) followed by culturing with SHH, FGF8, and CHIR99021, followed by neural differentiation in media containing BDNF, GDNF, TGFb3, and ascorbic acid (page 551, left col. para. 3; Figure 1d; Supplementary Figure 6). Kriks teaches the dopamine neurons derived from human hESCs engraft in vivo in Parkinson’s disease models including 6-hydroxy-dopamine-lesioned mice, rats, and parkinsonian monkeys (Abstract; page 549, left col. last para. and right col.; Figure 4; page 550). Kriks teaches the dopamine neurons completely restored amphetamine-induced rotation behavior and improved test of forelimb use and akinesia in mice and rats (Abstract). Kriks teaches excellent graft survival and behavioral outcome in the mouse and rat models but the number of dopamine neurons required in a mouse or rat brain represents only a fraction of the cells needed in a human (page 550, right col.). Kriks teaches the method was scalable producing batches of 5x107 dopamine neurons (page 550, right col.). Kriks teaches injecting 1.25x106 dopamine neurons each in six tracts of lesioned-rhesus monkeys and the neurons showed robust survival (page 550, right col.). Kriks teaches access to cells with the cardinal features of midbrain dopamine neurons will enable a broad range of biomedical applications and the method establishes a means for obtaining a scalable source of dopamine neurons for neural transplantation – a major step on the road towards considering a cell based therapy for Parkinson’s disease (page 551, left col. para. 2). Kriks does not teach “K02288”. One would have been motivated to combine the teachings of Neely, Hao, and Kriks to substitute 10 µM SB431542 in the method of Kriks with a lower concentration of DMH2 to make the method of producing large numbers of dopamine neurons for transplantation more economically affordable.
Regarding “K02288” of claims 52 and 57, Muguruma teaches a method of producing granule cells and deep cerebellar neurons (DCN) comprising contacting stem cells with K02288 (Figure 3E – I; Figure S3; page 542, right col. para. 1; Figure S3O and P; Figure 3I; page 544, left col. para. 1 – 2; page 549, right col. para. 4).
He teaches tremor is the most frequent initial motor symptom of Parkinson’s disease and the most difficult symptom to treat and the dentate nucleus may be involved in Parkinson’s disease tremor (Abstract; page 21, para. 3). The dentate nucleus is the largest deep cerebellar cluster of neurons as evidenced by de Leon (page 1, para. 1 – 2). He teaches current therapies for Parkinson’s disease only treat symptoms and do not reverse the degeneration in sub-cortical grey matter structures, which is particularly true for the treatment of Parkinson’s disease tremor that does not respond to dopamine replacement therapy (page 2, para. 2; page 7, last para.). He teaches Parkinson’s disease patients with tremor dominant (TD) phenotype have high iron concentrations within the dentate nucleus (page 8, para. 2). He teaches that it is possible that there is a reduction in dentate neurons due to elevated iron levels in these patients and this explanation accords well with functional and metabolic studies (page 8, para. 3). He teaches in contrast to TD phenotype, akinetic-rigid (AR) dominant Parkinson’s patients have increased dopaminergic neuronal loss (page 8, para. 5).
It would have been obvious prior to the effective filing date of the invention as claimed for the person of ordinary skill in the art to combine the teachings of Neely regarding a method of producing dopaminergic neurons by dual-SMAD inhibition followed by neural induction with the method comprising DMH1 with the teachings of Hao regarding DMH1 is a BMP signaling inhibitor but not a TGF-b1 signaling inhibitor and DMH2 is an inhibitor of BMP and TGF-b1 signaling with the teachings of Kriks regarding a method of producing large quantities of dopamine neurons using dual-SMAD inhibition followed by neural induction to treat Parkinson’s disease with the teachings of Murguruma regarding a method of producing DCN neurons with K02288 with the teachings of He regarding tremor-dominant Parkinson’s disease patients have dentate nucleus dysfunction to arrive at the claimed method where the compound comprises DMH1, DMH2, and K02288. One would have been motivated to combine the teachings of Neely, Hao, Kriks, Muguruma, and He in a cost-effective method of producing neurons for drug discovery and treatment of Parkinson’s disease patients with TD and AR phenotype as Neely teaches to ensure the successful use of hiPSC-derived neural precursors or differentiated neurons for cell-based therapy or drug discovery, standardized, chemically defined and economically affordable methods to derive these cells need to be developed and Kriks teaches excellent graft survival and behavioral outcome in the mouse and rat models but the number of dopamine neurons required in a mouse or rat brain represents only a fraction of the cells needed in a human and He teaches current therapies for Parkinson’s disease only treat symptoms and do not reverse the degeneration in sub-cortical grey matter structures and He teaches that it is possible that there is a reduction in dentate neurons due to elevated iron levels and He teaches Parkinson’s disease tremor does not respond to dopamine replacement therapy. One would have a reasonable expectation of success in combining the teachings as Neely teaches the method using DMH1 produces dopaminergic neurons and Hao teaches DMH2 is an inhibitor of BMP and TGF-b1 signaling and the IC50 for BMP inhibition by DMH2 is similar to that for LDN193189 (used in the method of Kriks) and Kriks teaches the dopamine neurons completely restored amphetamine-induced rotation behavior and improved test of forelimb use and akinesia in mice and rats (AR phenotype) and Muguruma teaches the method with K02288 produces deep cerebellar neurons (TD phenotype).
39. Claim(s) 1, 8, 13, 14, 19, 20, 21, 24, 31, 33, 47, 53, 58, 60 – 65, and 67 – 75 is/are rejected under 35 U.S.C. 103 as being unpatentable over Neely (Neely, M. Diana, et al. ACS chemical neuroscience 3.6 (2012): 482-491.), hereinafter Neely in view of Hao (Hao, Jijun, et al. ACS chemical biology 5.2 (2010): 245-253.), hereinafter Hao which is cited on the IDS filed 07/24/2023 in view of Kriks (Kriks, Sonja, et al. Nature 480.7378 (2011): 547-551.), hereinafter Kriks in view of Arshad (Valizadeh-Arshad, Zahra, et al. Cell Journal (Yakhteh) 19.4 (2017): 545.), hereinafter Arshad in view of Vogt (Vogt J, et. al. Ann Neurol. 2009 Sep;66(3):310-22.), hereinafter Vogt in view of Muguruma (Muguruma, Keiko, et al. Cell reports 10.4 (2015): 537-550.), hereinafter Muguruma in view of He (He N, et. al. NMR Biomed. 2017 Apr;30(4):10.1002/nbm.3554), hereinafter He as evidenced by de Leon (de Leon AS, Das JM. Neuroanatomy, Dentate Nucleus. [Updated 2023 Jul 24]. In: StatPearls [Internet]. Treasure Island (FL): StatPearls Publishing; 2025 Jan-. Available from: https://www.ncbi.nlm.nih.gov/books/NBK554381/), hereinafter de Leon.
Neely anticipates claims 1, 8, 13, 14, 19, 20, 21, 24, 31, 33, 47, 61 – 65, and 67 – 75 as set forth above. Neely teaches the compound comprises DMH1 (claim 58) but Neely does not teach the compound comprises A8301 and DMH2 of claim 53 or DMH1, DMH2, and A8301 of claim 58 or the compound comprises A8301, DMH2, and K0228 of claim 60. However, Neely teaches concomitant inhibition of the BMP and TGF-b1 branches of the TGF-b signaling pathways induces efficient neuralization of hiPSCs known as dual-SMAD inhibition (Abstract). Neely teaches DMH1 is a BMP-pathway inhibitor and SB431542 is a TGF-b1 inhibitor and treating hiPSCs with both compounds produces neural cells (Abstract; page 484, right col. last para; page 485, left col. para. 1). Neely teaches the method produces dopaminergic neurons where the neural induction media comprised DMH1 but not SB431542 (page 488, right col. para. 2 – 3; Figure 6; page 486, right col. last para.). Neely teaches to ensure the successful use of hiPSC-derived neural precursors or differentiated neurons for cell-based therapy or drug discovery, standardized, chemically defined and economically affordable methods to derive these cells need to be developed (page 483, right col. last para.; page 484, left col. para. 1; page 487, right col. para. 1). Neely teaches hiPSCs allow for the possibility of studying human neurons derived from patients with neurological diseases and efficient and reproducible methods to differentiate hiPSCs into neurons are critical to ensure successful use of these cells for the study of human developmental processes, cellular mechanisms underlying human disease, or the development of drug screening platforms (Abstract; page 487, right col. para. 1).
Regarding “DMH2” of claim 53, 58, and 60, Hao teaches in Table 2 that DMH1 is a BMP signaling inhibitor but not a TGF-b1 signaling inhibitor and DMH2 is an inhibitor of BMP and TGF-b1 signaling where the IC50 for TGF-b1 inhibition by DMH2 is 1578 nM. Hao teaches the IC50 for BMP inhibition (ALK2) by DMH2 (42.8 nM) is similar to that for LDN193189 (40.7 nM) in Table 2. Hao does not teach “A8301” of claims 53 and 60 or “K02288” of claim 60. One would have been motivated to combine the teachings of Neely and Hao to substitute 10 µM SB431542 in the method of Neely with a lower concentration of DMH2 to make the method of producing dopamine neurons more economically affordable.
Kriks teaches a method of deriving dopamine neurons from hESCs or hPSCs comprising dual-SMAD inhibition to accelerate neural fate acquisition with LDN193189 and SB43152 (10 µM) followed by culturing with SHH, FGF8, and CHIR99021, followed by neural differentiation in media containing BDNF, GDNF, TGFb3, and ascorbic acid (page 551, left col. para. 3; Figure 1d; Supplementary Figure 6). Kriks teaches the dopamine neurons derived from human hESCs engraft in vivo in Parkinson’s disease models including 6-hydroxy-dopamine-lesioned mice, rats, and parkinsonian monkeys (Abstract; page 549, left col. last para. and right col.; Figure 4; page 550). Kriks teaches the dopamine neurons completely restored amphetamine-induced rotation behavior and improved test of forelimb use and akinesia in mice and rats (Abstract). Kriks teaches excellent graft survival and behavioral outcome in the mouse and rat models but the number of dopamine neurons required in a mouse or rat brain represents only a fraction of the cells needed in a human (page 550, right col.). Kriks teaches the method was scalable producing batches of 5x107 dopamine neurons (page 550, right col.). Kriks teaches injecting 1.25x106 dopamine neurons each in six tracts of lesioned-rhesus monkeys and the neurons showed robust survival (page 550, right col.). Kriks teaches access to cells with the cardinal features of midbrain dopamine neurons will enable a broad range of biomedical applications and the method establishes a means for obtaining a scalable source of dopamine neurons for neural transplantation – a major step on the road towards considering a cell based therapy for Parkinson’s disease (page 551, left col. para. 2). Kriks does not teach “A8301” of claims 53 and 60 or “K02288” of claim 60. One would have been motivated to combine the teachings of Neely, Hao, and Kriks to substitute 10 µM SB431542 in the method of Kriks with a lower concentration of DMH2 to make the method of producing large numbers of dopamine neurons more economically affordable for cell-based therapy or drug discovery.
Regarding “A8301” of claim 53, 58, and 60, Arshad teaches a method of producing motor neurons comprising contacting stem cells with A8301 (page 546, left col. para. 3 – 4 and right col. para. 1; page 547, right col. para. 4 – 5; Figure 1). Arshad teaches motor neuron differentiation from hESCs is a goal of regenerative medicine to provide cell therapy as treatments for diseases that damage motor neurons (Abstract). Arshad does not teach “K02288” of claim 60. One would have been motivated to combine the teachings of Neely, Hao, Kriks, and Arshad in an economically affordable method of producing neurons for cell-based therapy or drug discovery.
Vogt teaches MS patients have a massive loss of lower motor neurons (Abstract; page 312, right col. para. 2; Figure 1E, F). Vogt teaches spinal motor neurons in chronic active MS exhibited morphological signs of apoptosis (page 312, right col. para. 2). Vogt teaches the assumption that MS is a purely demyelinating disease has been challenged by the observation that axonal damage is a common finding in the CNS of MS patients (page 310, left col. and right col. para. 1). Vogt teaches because the cell death of lower motor neurons plays a prominent role in MS pathology, there is a reason to pursue scientific efforts to develop novel therapeutics strategies with neuroprotective potential (page 321, left col. para. 3). Vogt does not teach “K02288”. One would have been motivated to combine the teachings of Petersen, Arshad, and Vogt because Petersen and Arshad teach methods of treating the effects observed in MS lesions regarding remyelination failure and lack of motor neurons. Vogt does not teach “K02288” of claim 60. One would have been motivated to combine the teachings of Neely, Hao, Kriks, Arshad, and Vogt in an economically affordable method of producing neurons for cell-based therapy or drug discovery.
Regarding “K02288” of claim 60, Muguruma teaches a method of producing granule cells and deep cerebellar neurons (DCN) comprising contacting stem cells with K02288 (Figure 3E – I; Figure S3; page 542, right col. para. 1; Figure S3O and P; Figure 3I; page 544, left col. para. 1 – 2; page 549, right col. para. 4).
He teaches tremor is the most frequent initial motor symptom of Parkinson’s disease and the most difficult symptom to treat and the dentate nucleus may be involved in Parkinson’s disease tremor (Abstract; page 21, para. 3). The dentate nucleus is the largest deep cerebellar cluster of neurons as evidenced by de Leon (page 1, para. 1 – 2). He teaches current therapies for Parkinson’s disease only treat symptoms and do not reverse the degeneration in sub-cortical grey matter structures, which is particularly true for the treatment of Parkinson’s disease tremor that does not respond to dopamine replacement therapy (page 2, para. 2; page 7, last para.). He teaches Parkinson’s disease patients with tremor dominant (TD) phenotype have high iron concentrations within the dentate nucleus (page 8, para. 2). He teaches that it is possible that there is a reduction in dentate neurons due to elevated iron levels in these patients and this explanation accords well with functional and metabolic studies (page 8, para. 3). He teaches in contrast to TD phenotype, akinetic-rigid (AR) dominant Parkinson’s patients have increased dopaminergic neuronal loss (page 8, para. 5).
It would have been obvious prior to the effective filing date of the invention as claimed for the person of ordinary skill in the art to combine the teachings of Neely regarding a method of producing neurons comprising DMH1 with the teachings of Hao regarding DMH1 is a BMP signaling inhibitor but not a TGF-b1 signaling inhibitor and DMH2 is an inhibitor of BMP and TGF-b1 signaling with the teachings of Kriks regarding a scalable method of producing neurons for treating Parkinson’s disease with the teachings of Arshad regarding a method of producing motor neurons with A8301 with the teachings of Vogt regarding MS patients have fewer motor neurons with the teachings of Muguruma regarding a method of producing deep cerebellar neurons using K02288 with the teachings of He regarding tremor-dominant Parkinson’s disease patients have dentate nucleus dysfunction to arrive at the claimed method where the compound comprises DMH1, DMH2, K02288, and A8301. One would have been motivated to combine the teachings of Neely, Hao, Kriks, Arshad, Vogt, Muguruma, and He in a cost-effective method produce neurons for drug discovery and therapy as Neely teaches to ensure the successful use of hiPSC-derived neural precursors or differentiated neurons for cell-based therapy or drug discovery, standardized, chemically defined and economically affordable methods to derive these cells need to be developed and Neely teaches hiPSCs allow for the possibility of studying human neurons derived from patients with neurological diseases and efficient and reproducible methods to differentiate hiPSCs into neurons are critical to ensure successful use of these cells for the study of human developmental processes, cellular mechanisms underlying human disease, or the development of drug screening platforms and Kriks teaches excellent graft survival and behavioral outcome in the mouse and rat models but the number of dopamine neurons required in a mouse or rat brain represents only a fraction of the cells needed in a human and Arshad teaches motor neuron differentiation from hESCs is a goal of regenerative medicine to provide cell therapy as treatments for diseases that damage motor neurons and Vogt teaches because the cell death of lower motor neurons plays a prominent role in MS pathology, there is a reason to pursue scientific efforts to develop novel therapeutics strategies with neuroprotective potential and He teaches current therapies for Parkinson’s disease only treat symptoms and do not reverse the degeneration in sub-cortical grey matter structures. One would have a reasonable expectation of success in combining the teachings as Neely teaches the method using DMH1 produces dopaminergic neurons and Hao teaches DMH2 is an inhibitor of BMP and TGF-b1 signaling and the IC50 for BMP inhibition by DMH2 is similar to that for LDN193189 (used in the method of Kriks) and Kriks teaches the dopamine neurons completely restored amphetamine-induced rotation behavior and improved test of forelimb use and akinesia in mice and rats (AR phenotype) and Arshad teaches the method using A8031 produces motor neurons and Muguruma teaches the method using K02288 produces deep cerebellar neurons.
40. Claim(s) 1, 14, 28, 31, 33, 47, 59, and 63 – 65 is/are rejected under 35 U.S.C. 103 as being unpatentable over Petersen (Petersen, Mark A., et al. Neuron 96.5 (2017): 1003-1012.), hereinafter Petersen in view of Arshad (Valizadeh-Arshad, Zahra, et al. Cell Journal (Yakhteh) 19.4 (2017): 545.), hereinafter Arshad in view of Vogt (Vogt J, et. al. Ann Neurol. 2009 Sep;66(3):310-22.), hereinafter Vogt in view of Muguruma (Muguruma, Keiko, et al. Cell reports 10.4 (2015): 537-550.), hereinafter Muguruma in view of Albert (Albert, Monika, et al. Brain Pathology 27.6 (2017): 737-747.), hereinafter Albert as evidenced by de Leon (de Leon AS, Das JM. Neuroanatomy, Dentate Nucleus. [Updated 2023 Jul 24]. In: StatPearls [Internet]. Treasure Island (FL): StatPearls Publishing; 2025 Jan-. Available from: https://www.ncbi.nlm.nih.gov/books/NBK554381/), hereinafter de Leon.
Petersen anticipates claims 1, 14, 28, 31, 33, 47, and 63 – 65 as set forth above. Petersen does not teach the compound comprises DMH1, A8301, and K02288 of claim 59. However, Petersen teaches fibrinogen activates BMP signaling pathway in oligodendrocyte progenitor cells (OPCs) and suppresses remyelination (Summary; page 1008, left col. last para. and right col. para. 2; Figure 4E). Petersen teaches fibrinogen induces phosphorylation of Smad 1/5/8 and inhibits OPC differentiation into myelinating oligodendrocytes and the effects of fibrinogen are rescued by BMP type I receptor inhibition using DMH1 (Summary; page 1005, left col. para. 3; page 1005, right col. para. 2; Figure 2D, E). Petersen teaches the BMP target Id2 is increased in demyelinated multiple sclerosis (MS) lesions and therapeutic depletion of fibrinogen decreases BMP signaling and enhances remyelination in vivo (Summary). Petersen teaches targeting fibrinogen may be an upstream therapeutic strategy to promote the regenerative potential of CNS progenitors in diseases with remyelination failure (Summary). Petersen teaches remyelination is critical for recovery in several diseases such as MS and OPCs often fail to differentiate into mature oligodendrocytes required for myelin repair (page 1004, right col.). Petersen teaches fibrinogen is deposited in MS, stroke, traumatic brain injury, and Alzheimer’s disease and fibrinogen is found in active and chronic MS lesions (page 1005, left col. para. 2). Petersen teaches fibrinogen is a driver of neuropathology promoting neuroinflammation and glial scar formation by direct effects on microglia, astrocytes, and neurons (page 1005, left col. para. 2). Petersen teaches fibrinogen increased expression of several BMP-responsive genes including Id1, Id2, Nog, Hes1, Hey1, and Lef1 that are associated with impaired OPC differentiation and upregulated in some MS patients (page 1005, right col. para. 1; Figure 1F). Petersen teaches active lesions of MS patients had high levels of Id2 in areas of fibrinogen deposition and demyelination (page 1005, right col. para. 1). Petersen teaches DMH1 is a dorsomorphin analog that inhibits ACVR1 (Alk2) and it blocked fibrinogen-induced phosphorylation of Smad1/5 and suppressed the Id genes in cultures of OPCs (page 1005, right col. para. 2; Figure 2D, E). Petersen teaches DMH1 increased the number of mature oligodendrocytes (page 1008, left col. para. 1). Petersen teaches depletion of fibrinogen in vivo reduced phospho-Smad 1/5/8 in demyelinated lesions suggesting reduction of BMP receptor pathway activation concomitant with increased mature oligodendrocytes and enhanced remyelination (page 1008, left col. para. 2).
Regarding “A8301”, Arshad teaches a method of producing motor neurons comprising contacting stem cells with A8301 (page 546, left col. para. 3 – 4 and right col. para. 1; page 547, right col. para. 4 – 5; Figure 1). Arshad teaches motor neuron differentiation from hESCs is a goal of regenerative medicine to provide cell therapy as treatments for diseases that damage motor neurons (Abstract). Arshad does not teach “K02288”.
Vogt teaches MS patients have a massive loss of lower motor neurons (Abstract; page 312, right col. para. 2; Figure 1E, F). Vogt teaches spinal motor neurons in chronic active MS exhibited morphological signs of apoptosis (page 312, right col. para. 2). Vogt teaches the assumption that MS is a purely demyelinating disease has been challenged by the observation that axonal damage is a common finding in the CNS of MS patients (page 310, left col. and right col. para. 1). Vogt teaches because the cell death of lower motor neurons plays a prominent role in MS pathology, there is a reason to pursue scientific efforts to develop novel therapeutics strategies with neuroprotective potential (page 321, left col. para. 3). Vogt does not teach “K02288”. One would have been motivated to combine the teachings of Petersen, Arshad, and Vogt because Petersen and Arshad teach methods of treating the effects observed in MS lesions regarding remyelination failure and lack of motor neurons.
Regarding “K02288”, Muguruma teaches a method of producing granule cells and deep cerebellar neurons (DCN) comprising contacting stem cells with K02288 (Figure 3E – I; Figure S3; page 542, right col. para. 1; Figure S3O and P; Figure 3I; page 544, left col. para. 1 – 2; page 549, right col. para. 4). Muguruma teaches K0228 is a BMP inhibitor (page S6).
Albert teaches the dentate nucleus is the predilection site for lesion development in MS (Abstract; page 737, left col. para. 1). Albert teaches a significant reduction of synapses in the dentate nucleus of MS patients with or without the presence of demyelination (Abstract; page 741; Figure 1; page 742, left col. para. 1 and last para.; page 746, left col. para. 3). Albert teaches neuronal loss in the dentate nucleus was more pronounced in cases with local demyelination (page 743, right col.). The dentate nucleus is the largest deep cerebellar cluster of neurons as evidenced by de Leon (page 1, para. 1 – 2).
It would have been obvious prior to the effective filing date of the invention as claimed for the person of ordinary skill in the art to combine the teachings of Petersen regarding a method of producing oligodendrocytes from OPCs by rescuing the effects of fibrinogen with DMH1 with the teachings of Arshad regarding a method of producing motor neurons by contacting stem cells with A8301 with the teachings of Vogt regarding MS patients have fewer motor neurons with the teachings of Muguruma regarding a method of producing deep cerebellar neurons by contacting stem cells with K02288 with the teachings of Albert regarding MS patients have reduced synapses in the dentate nucleus to arrive at the claimed method where the compound comprises DMH1, A8301, and K02288. One would have been motivated to combine the teachings of Petersen, Arshad, Vogt, Muguruma, and Albert in a method to treat multiple sclerosis with oligodendrocytes, motor neurons, and deep cerebellar neurons as Petersen teaches targeting fibrinogen may be an upstream therapeutic strategy to promote the regenerative potential of CNS progenitors in diseases with remyelination failure and Petersen teaches remyelination is critical for recovery in several diseases such as MS and OPCs often fail to differentiate into mature oligodendrocytes required for myelin repair and Petersen teaches fibrinogen is deposited in MS, found in active and chronic MS lesions, and is a driver of neuropathology promoting neuroinflammation and glial scar formation by direct effects on microglia, astrocytes, and neurons and Vogt teaches because the cell death of lower motor neurons plays a prominent role in MS pathology, there is a reason to pursue scientific efforts to develop novel therapeutics strategies with neuroprotective potential and Albert teaches neuronal loss in the dentate nucleus was more pronounced in cases with local demyelination. One would have a reasonable expectation of success in combining the teachings as Petersen teaches decreasing BMP signaling enhances remyelination in vivo and DMH1 blocks the effects of fibrinogen and Arshad teaches A8301 treatment produces motor neurons and Muguruma teaches K02288 treatment produces deep cerebellar neurons and Muguruma teaches K02288 is a BMP inhibitor.
Double Patenting
The nonstatutory double patenting rejection is based on a judicially created doctrine grounded in public policy (a policy reflected in the statute) so as to prevent the unjustified or improper timewise extension of the “right to exclude” granted by a patent and to prevent possible harassment by multiple assignees. A nonstatutory double patenting rejection is appropriate where the conflicting claims are not identical, but at least one examined application claim is not patentably distinct from the reference claim(s) because the examined application claim is either anticipated by, or would have been obvious over, the reference claim(s). See, e.g., In re Berg, 140 F.3d 1428, 46 USPQ2d 1226 (Fed. Cir. 1998); In re Goodman, 11 F.3d 1046, 29 USPQ2d 2010 (Fed. Cir. 1993); In re Longi, 759 F.2d 887, 225 USPQ 645 (Fed. Cir. 1985); In re Van Ornum, 686 F.2d 937, 214 USPQ 761 (CCPA 1982); In re Vogel, 422 F.2d 438, 164 USPQ 619 (CCPA 1970); In re Thorington, 418 F.2d 528, 163 USPQ 644 (CCPA 1969).
A timely filed terminal disclaimer in compliance with 37 CFR 1.321(c) or 1.321(d) may be used to overcome an actual or provisional rejection based on nonstatutory double patenting provided the reference application or patent either is shown to be commonly owned with the examined application, or claims an invention made as a result of activities undertaken within the scope of a joint research agreement. See MPEP § 717.02 for applications subject to examination under the first inventor to file provisions of the AIA as explained in MPEP § 2159. See MPEP § 2146 et seq. for applications not subject to examination under the first inventor to file provisions of the AIA . A terminal disclaimer must be signed in compliance with 37 CFR 1.321(b).
The filing of a terminal disclaimer by itself is not a complete reply to a nonstatutory double patenting (NSDP) rejection. A complete reply requires that the terminal disclaimer be accompanied by a reply requesting reconsideration of the prior Office action. Even where the NSDP rejection is provisional the reply must be complete. See MPEP § 804, subsection I.B.1. For a reply to a non-final Office action, see 37 CFR 1.111(a). For a reply to final Office action, see 37 CFR 1.113(c). A request for reconsideration while not provided for in 37 CFR 1.113(c) may be filed after final for consideration. See MPEP §§ 706.07(e) and 714.13.
The USPTO Internet website contains terminal disclaimer forms which may be used. Please visit www.uspto.gov/patent/patents-forms. The actual filing date of the application in which the form is filed determines what form (e.g., PTO/SB/25, PTO/SB/26, PTO/AIA /25, or PTO/AIA /26) should be used. A web-based eTerminal Disclaimer may be filled out completely online using web-screens. An eTerminal Disclaimer that meets all requirements is auto-processed and approved immediately upon submission. For more information about eTerminal Disclaimers, refer to www.uspto.gov/patents/apply/applying-online/eterminal-disclaimer.
41. Claim 1, 8, 10, 11, 13 – 15, 19 – 21, 24, 25, 28, 31, 33, 35, 36, and 47 – 75 are provisionally rejected on the ground of nonstatutory double patenting as being unpatentable over claim 1, 2, 9, 11, 16, 19, 29, 30, 33, 34, 38, 42, 51, 54, 55, 62, 67, 69 of copending Application No. 18855867 (reference application). Although the claims at issue are not identical, they are not patentably distinct from each other because instant claim 1 is anticipated by reference claim 1 and 2.
Instant claim 1 is recites a method for differentiating stem or progenitor cells into neural cells or for producing neural cells from stem or progenitor cells, the method comprising contacting the cells with a compound selected from DMH1, DMH2, K02288, A8301, or combinations thereof.
Reference claim 1 recites a method for differentiating stem or progenitor cells into neural cells, the method comprising (i) contacting the stem or progenitor cells with a differentiation composition, wherein the differentiation composition comprises one or more of the ALK inhibitors: DMH1, DMH2, K02288, and A83-0l; and (ii) culturing the cells in microwells to form spheroids and/or neurospheres. Reference claim 2 recites a method for differentiating stem or progenitor cells into neural cells, the method comprising (i) contacting the stem or progenitor cells with a differentiation composition; and (ii) culturing the cells in microwells to form spheroids and/or neurospheres. Therefore reference claims 1 and 2 are in essence a “species” of the generic invention of instant claim 1. It has been held that a generic invention is “anticipated” by a “species” within the scope of the generic invention. See In re Goodman, 29 USPQ2d 2010 (Fed. Cir. 1993).
This is a provisional nonstatutory double patenting rejection because the patentably indistinct claims have not in fact been patented.
Conclusion
No claims allowed.
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/Z.M.B./Examiner, Art Unit 1632
/MARCIA S NOBLE/Primary Examiner, Art Unit 1632