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 .
Election/Restrictions
Applicant’s election with traverse of the following species in the reply filed on December 17, 2025, is acknowledged.
Applicant elected the following species:
a. Immune cells
Furthermore, Applicant did not identify which species of method (claims 1-16 and 20 or 17-19) were elected in the response to the election mailed on August 26, 2025. However, Applicant provisionally elected the method of claims 1-16 and 20 in a telephonic interview with their representative, Peter D. Weinstein, on February 10, 2026.
Claims 17-18 are withdrawn from further consideration pursuant to 37 CFR 1.142(b) as being drawn to a nonelected inventions or species, there being no allowable generic or linking claim.
Although Applicant made the Species election with traverse, Applicant did not distinctly and specifically point out the supposed errors in the restriction requirement. As such, the election has been treated as an election without traverse (MPEP § 818.01(a)).
DETAILED ACTION
The amended claims filed on December 17, 2025, have been acknowledged. Claims 7-8 and 19 were cancelled. Claims 1, 9, and 17 were amended. In light of the Applicant’s elected species, claims 17-18 are withdrawn from further consideration pursuant to 37 CFR 1.142(b) as being drawn to a nonelected invention, there being no allowable generic or linking claim. Claims 1-6, 9-16, and 20 are pending and examined on the merits.
Priority
The applicant claims domestic priority from U.S. provisional application No. 63/170,885, filed on April 5, 2021. Applicant’s claim for the benefit of a prior-filed application under 35 U.S.C. 119(e) or under 35 U.S.C. 120, 121, 365(c), or 386(c) is acknowledged. Claims 1-6, 9-16, and 20 receive domestic benefit from U.S. provisional application No. 63/170,885, filed on April 5, 2021.
Specification
The use of the term benzonase, 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 Rejections - 35 USC § 112
The following is a quotation of the first paragraph of 35 U.S.C. 112(a):
(a) IN GENERAL.—The specification shall contain a written description of the invention, and of the manner and process of making and using it, in such full, clear, concise, and exact terms as to enable any person skilled in the art to which it pertains, or with which it is most nearly connected, to make and use the same, and shall set forth the best mode contemplated by the inventor or joint inventor of carrying out the invention.
The following is a quotation of the first paragraph of pre-AIA 35 U.S.C. 112:
The specification shall contain a written description of the invention, and of the manner and process of making and using it, in such full, clear, concise, and exact terms as to enable any person skilled in the art to which it pertains, or with which it is most nearly connected, to make and use the same, and shall set forth the best mode contemplated by the inventor of carrying out his invention.
Claims 1-6, 9-16, and 20 are rejected under 35 U.S.C. 112(a) or 35 U.S.C. 112 (pre-AIA ), first paragraph, because while the prior art provides enablement for treating T cells with benzonase to generate genetically modified T cells expressing TCRs to treat cancer or treating cryopreserved leukocytes and platelets with benzonase after thawing and administering the cell to cancer patients to treat thrombocytopenia, does not reasonably provide enablement for treating and administering any immune cell using any of the claimed treatments and administering these cells to treat any cancer. The specification does not enable any person skilled in the art to which it pertains, or with which it is most nearly connected, to use the invention commensurate in scope with these claims.
The factors to be weighed to evaluate whether a disclosure satisfies the enablement requirement and whether any necessary experimentation is undue are set forth in MPEP 2164.01(a).
(A) The breadth of the claims;
(B) The nature of the invention;
(C) The state of the prior art;
(D) The level of one of ordinary skill;
(E) The level of predictability in the art;
(F) The amount of direction provided by the inventor;
(G) The existence of working examples; and
(H) The quantity of experimentation needed to make or use the invention based on the content of the disclosure.
Although all the factors have been considered, the relevant factors will be addressed below.
Breadth of the claims: Claim 1 recites the following claim language, “A method for treating a cancer, wherein
a. an immune cell, which is not a chimeric antigen receptor T-cell, is treated with one or more of an RNase, a DNase, an enzyme that has DNase and/or RNase activity, an antibody that binds to an RNA and an antibody that binds to a DNA;
b. following treatment, a therapeutically effective amount of the treated cells are administered to a patient suffering from a cancer; and
c. following administration, the treated cells reduce the symptoms of the cancer”.
The broadest reasonable interpretation is that this method could be used with any immune cell treated with any RNase, DNase, enzyme that has DNase and/or RNase activity, antibody that binds to an RNA, and antibody that binds to a DNA at any concentration through administration of the treated cell through any administration route to treat any cancer.
Nature of the invention: The subject matter of the invention relates to a method of treating cancer by treating immune cell with a compound that breaks down DNA and RNA prior to administering he cells to a patient.
State of the prior art: The prior art teaches that T cells can be treated with benzonase as part of a method of transducing T cells with a TCR and administering these cells to patients with cancer, as identified by United States Patent Application No. 20190247433 (Kalra) and Yang et al. (Human Gene Therapy Methods 23: 73-83. 2012).
Kalra teaches a method of transducing T cells with TCRs, comprising:
Frozen PBMC from healthy donors were thawed in warm media supplemented
with 5% human AB serum. Cells were treated with benzonase nuclease (50 U/ml) for 15 minutes at 37° C., washed, counted, and put to overnight rest in complete
media.
On a day when cells are thawed, 24-well non-tissue culture plates were coated with anti-CD3 and anti-CD28 antibodies diluted in PBS (1 μg/mL), sealed and incubated
overnight at 4° C. Next day, rested PBMCs were harvested, counted, washed and resuspended at the concentration of 1x106/ml. Antibody solution was aspirated, and wells were washed with complete media followed by addition of 2 x106 cells to each well. Activation was carried out at 37° C.
Activated T cells were harvested, washed and counted. Transduction mixtures containing concentrated virus supernatants, protamine sulfate (10 μg/ml), IL-7 (10
ng/ml) and IL-15 (100 ng/ml) were prepared. For each transduction, 1x106 cells were separated in a sterile microcentrifuge tube and centrifuged at 400xg for 5 minutes. Each
cell pellet was resuspended in 0.5 ml of the transduction mixture corresponding to a specific MOI. Cell suspension was placed in an appropriately labelled well of a 24-well
G-Rex plate. After 24 hours of incubation at 37° C. and 5% CO2 , 1.5 ml media supplemented with IL-7 (10 ng/ml) and IL-15 (100 ng/ml) was added to each well. Ninety-six-hour post-transduction, transgene expression was determined by
flow cytometry. Multimeric MHC-peptide complexes (Dextramer or Tetramer) were used to monitor surface expression of transgenic TCR by FACS.
The method of Kalra achieved successful transduction with each of two different lentiviral constructs encoding R7Pl D5 TCR, i.e., LV-R73 and LV-R78 (Example 2 and Figure 14.
Kalra teaches that their TCR expressing T cells can be used to treat cancer by reducing the size of a tumor or preventing the growth or re-growth of a tumor in these subjects (paragraphs 0159-0169).
Yang teaches a method of transducing peripheral blood lymphocytes (including T cells) with a MART-1 TCR to target melanoma, comprising:
For lentiviral vector production, Vesicular stomatitis virus glycoprotein (VSV-G)-
pseudotyped lentiviral vector was produced transiently by transfection, using calcium phosphate and a four-plasmid system. The concentrated vector was treated with Benzonase (50 U/ml) for 1 hr at room temperature followed by diafiltration with 5 volumes (10 liters) of AIM-V medium. The final product was aliquoted into 50-ml Cryocyte bags and stored at - 80°C until further use.
For PBL transduction, On day 0, PBLs (5 x 106 cell/ml, 2 ml/well) were stimulated with interleukin (IL)-2 (300 IU/ml) and anti-CD3/anti-CD28 beads (ratio of beads to cells was 3:1) in tissue culture-treated 6-well plates and incubated overnight at 37°C and 5% CO2. Serial dilutions of vector supernatant containing protamine sulfate (10 lg/ml) were applied to 1 x 106 stimulated PBLs (final volume, 1 ml) in individual wells of a 24-well plate. The plates were centrifuged at 2000 · g for 2 hr at 32°C followed by incubation at 37°C and 5% CO2. The next day, the medium was replaced with AIM-V medium containing 5% human serum and IL-2 (300 IU/ml), and the cells were transferred to 25-cm2 tissue culture flasks at a density of 0.3 x 106 cells/ml.
The current World Health Organization (WHO) regulatory guidance limit for residual DNA in biological products derived from continuous cell lines is 10 ng/dose. The concentration of residual plasmid DNA in the vector supernatant after Benzonase treatment was below the detection limit of the assay (2.0 ng of DNA per milliliter;
Supplementary Table S1) whereas without the benzonase treatment, the residual DNA was ~12 ng/mL, above the guidance limit.
Yang identifies that treating the lentiviral supernatant with benzonase leads to residual benzonase during transduction of the PBLs and does not reduce PBL transduction efficiency or function, leading to expression of the TCR in the PBLs (Figure 3).
The prior art teaches that transfusion of cryopreserved platelets that are leukocyte reduced can be used to treat thrombocytopenia associated with neoplasia and that benzonase can be used during thawing cryopreserved PBMCs to remove residual cell free DNA, as identified by Vadhan-Raj et al. (The Lancet 359: 2145-2152. 2002), Federici et al. (Thrombosis Research 129: S60-S65. 2012), Chen et al. (BMC Immunology 21: 1-13. 2020), and Fry et al. (Thesis: Cell Separation and Cryopreservation of Cord Blood Fractions for Immunotherapeutic Applications. 2014).
Vadhan-Raj teaches that cryopreserved platelets can be administered to cancer patients experiencing thrombocytopenia (abstract)
Federici teaches that platelets can be administered to cancer patients experiencing thrombocytopenia associated with neoplasia. As part of the platelet transfusion, platelets are collected through apheresis procedures which leads to a leukocyte-reduced (but not eliminated) platelet concentrate (abstract and page S62, column 1, paragraph 2- page S63, column 1, paragraph 1).
Chen teaches that benzonase can be added to isolated cryopreserved cells from the blood after they have thawed (page 11, column 1, paragraph 2).
Fry teaches that DNase is often used to break down cell free DNA after thawing bone marrow cells (page 22, paragraph 2).
Level of predictability in the art: The prior art has successfully reduced to practice that T cells can be treated with benzonase at two different stages during manufacturing of T cells transduced with TCRs. Furthermore, the prior art has shown that leukocyte reduced platelets can be administered to cancer patients to treat thrombocytopenia and that benzonase/DNase can be used to remove residual cell free DNA after thawing
As such, this results in unpredictability about how someone can use any of the other compounds with DNase/RNase activity (enzymes with DNase/RNase activity and antibodies that bind to RNA or DNA) and use this method to treat any other immune cell that are not T cells or platelets and leukocytes.
Amount of direction provided by the inventor and existence of working examples: The specification discloses examples 1-7 which focus on treating cells with a DNase and/or RNase to improve cancer treatment efficacy. As an initial matter, the specification routinely identifies that they used SL4 cells for treating cancer in their examples, but does not definitively identify what the SL4 cells are. As outlined below, it is not clear what these SL4 cells are for multiple reasons.
In example 1, the specification discloses that granulocytes and agranulocytes were treated with an RNase and a DNase and then Myeloperoxidase (MPO), Neutrophil Acid Phosphatase (NACP), and Neutrophil Alkaline Phosphatase (NAP) activities were measured. Table 3 compares SL4-C and SL4-T cells for MPO, NAP, and NAcP activity. Therefore, the SL4 cells could be granulocytes alone, agranulocytes alone, or granulocytes and agranulocytes in combination.
Furthermore, Example 2 of the specification discloses that SL4-C (presumably untreated SL4 cells as this nomenclature is also not defined) and SL4-T (presumably treated SL4 cells as this nomenclature is also not defined) cells are derived from whole blood, buffy coats, leukapheresis, Filgrastim or Plerixafor. Therefore, SL4 cells could be any cell collected through these methods.
Additionally, in Example 2 of the specification, the specification states results showed SL4-T cells, significantly enhanced anticancer activity against tumor cells, with Table 4 following this statement. Table 4 of example 2 provides a multitude of possible cells and cell combinations that could be SL4 cells
Furthermore, the art identifies SL4 cells as mouse colon carcinoma cells and not immune cells (Morimoto-Tomita (Clinical & Experimental Metastasis 22: 513–521. 2005)).
Example 3 only identifies administering SL4 cells to treat cancer without identifying which of the individual or combination of cells are the SL4 cells. Only in Table 7 are specific administered cell types identified with SL21-Tn and SL21-Ta identifying that granulocytes, agaranulocytes and platelets are part of the composition. However, SL21 is not defined. Therefore, it is not clear whether these are the SL21 cells or if another cell type is part of the SL21 composition.
Examples 4 and 6-7 discloses multiple patients treated with SL4 cells, but does not identify the cells encompassed within the SL4 composition.
Example 5 focuses on treating CD19 CAR T-cells and is specifically identified as not being within the scope of claim 1.
As the SL4 cells were derived from whole blood, buffy coats, leukapheresis, Filgrastim, or Plerixafor this leads to the conclusion that the only viable cells for this type of treatment for cancer would be cells present in whole blood. Further Filgrastim is a medication used to treat neutropenia comprised of granulocyte stimulating factor (G-CSF) and Plerixafor is another medication used to stimulate stem cell mobilization from the bone marrow to the peripheral blood by antagonizing the CXCR4 receptor, so how any cell could be derived therefrom is unclear (Flomenberg, Biol Blood Marrow Transplant. 2010 May;16(5):695-700 and Dale, Support Care Cancer. 2018 Jan;26(1 ):7-20).
Based on all of the above, it is unclear which cells are encompassed by the undefined SL4 cells and which cells/combination of cells are used throughout the Applicant’s examples.
Furthermore, regarding the treatment, none of the examples clearly define the treatment method. Example 1 identifies that granulocytes and agranulocytes were treated with an RNase and a DNase but doesn’t identify whether it was a specific RNase/DNase nor the concentration or the number of times it was applied/how long it was applied.
Similarly, Example 2, discloses that activation of WBCs utilized a different DNase, RNase, including S1 and Micrococcal nuclease to target G-quadruplexes and ZONA but only identifies that S1 nuclease and Micrococcal were used to treat Granulocytes+Agranulocytes+platelets and does not identify the concentration or the number of times it was applied/how long it was applied to the cells.
Example 3 only identifies that granulocytes, agaranulocytes and platelets treated with nucleases or antinucleic antibodies as part of the SL21 composition.
Examples 4 and 6-7 provide no information on the treatment used for the SL4 cells prior to administration to the patients,
As such, the exact method of treating the cells is unclear, including what nuclease/antinucleic antibody is being used, what concentration is being used, how many applications, and how long treatment occurs. Furthermore, it is not clear whether the nucleases/antinucleic acids are removed prior to administering the cells or whether they are administered with the cells.
Quantity of experimentation needed: In light of the above factors, the prior art provides enablement for treating T cells with benzonase to generate genetically modified T cells expressing TCRs to treat cancer or treating cryopreserved leukocytes and platelets with benzonase after thawing and administering the cell to cancer patients to treat thrombocytopenia, but does not reasonably provide enablement for treating and administering any immune cell using any of the claimed treatments to treat any cancer. As identified above, the lack of clarity regarding the SL4 cells and treatments renders the information in Applicant’s examples non-enabling. As such, there would be undue experimentation related to using any other immune cell and any other treatment not identified in the prior art to practice the full scope of the claim.
Claims 2-6, 9-16, and 20 are also rejected because of their dependency on claim 1.
The following is a quotation of 35 U.S.C. 112(b):
(b) CONCLUSION.—The specification shall conclude with one or more claims particularly pointing out and distinctly claiming the subject matter which the inventor or a joint inventor regards as the invention.
The following is a quotation of 35 U.S.C. 112 (pre-AIA ), second paragraph:
The specification shall conclude with one or more claims particularly pointing out and distinctly claiming the subject matter which the applicant regards as his invention.
Claims 13-14 and 16 are rejected under 35 U.S.C. 112(b) or 35 U.S.C. 112 (pre-AIA ), second paragraph, as being indefinite for failing to particularly point out and distinctly claim the subject matter which the inventor or a joint inventor (or for applications subject to pre-AIA 35 U.S.C. 112, the applicant), regards as the invention.
Claim 13 contains the trademark/trade name benzonase. 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 Serratia marcescens endonuclease and, accordingly, the identification/description is indefinite.
Claim 14 recites the limitation "wherein the two or more treatments" in line 1. There is insufficient antecedent basis for this limitation in the claim. Claim 14 is dependent on claim 11 which is dependent on claim 1. Neither claim identifies treating the cells two or more times. Instead claim 12 discusses treating the cells two or more times. It is recommended that Applicant amends claim 14 to be dependent on claim 12 instead of claim 11.
Claims 16 recites the limitation “epidural and infusion” in line 4. It is not clear whether epidural and infusion are meant to be one route of administration, or is epidural and infusion are separate routes of administration as an epidural can involve a single injection or continuous infusion. Due to the lack of clarity, epidural and infusion are considered separate routes of administration.
The following is a quotation of 35 U.S.C. 112(d):
(d) REFERENCE IN DEPENDENT FORMS.—Subject to subsection (e), a claim in dependent form shall contain a reference to a claim previously set forth and then specify a further limitation of the subject matter claimed. A claim in dependent form shall be construed to incorporate by reference all the limitations of the claim to which it refers.
The following is a quotation of pre-AIA 35 U.S.C. 112, fourth paragraph:
Subject to the following paragraph [i.e., the fifth paragraph of pre-AIA 35 U.S.C. 112], a claim in dependent form shall contain a reference to a claim previously set forth and then specify a further limitation of the subject matter claimed. A claim in dependent form shall be construed to incorporate by reference all the limitations of the claim to which it refers.
Claim 9 is rejected under 35 U.S.C. 112(d) or pre-AIA 35 U.S.C. 112, 4th paragraph, as being of improper dependent form for failing to further limit the subject matter of the claim upon which it depends, or for failing to include all the limitations of the claim upon which it depends.
Claim 9 recites “The immune cells of claim 1”. However, claim 1 is a method. Claim 1 should be amended to state “the method of claim 1, wherein the immune cells are selected from …”. See claim 6 as an example of proper claim construction.
Applicant may cancel the claim(s), amend the claim(s) to place the claim(s) in proper dependent form, rewrite the claim(s) in independent form, or present a sufficient showing that the dependent claim(s) complies with the statutory requirements.
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.
Claims 1-4, 6, 9-10, 13, and 15-16 is/are rejected under 35 U.S.C. 102(a)(1) and 102(a)(2) as being anticipated by United States Patent Application No. 20190247433 (Kalra).
Regarding claims 1-4, 9, and 13, Kalra teaches a method of transducing T cells with TCRs (a bioproduct with anticancer activity), comprising:
Frozen PBMCs from healthy donors were thawed in warm media supplemented
with 5% human AB serum. Cells were treated with benzonase nuclease (50 U/ml) for 15 minutes at 37° C., washed, counted, and put to overnight rest in complete
media.
On a day when cells are thawed, 24-well non-tissue culture plates were coated with anti-CD3 and anti-CD28 antibodies diluted in PBS (1 μg/mL), sealed and incubated
overnight at 4° C. Next day, rested PBMCs were harvested, counted, washed and resuspended at the concentration of 1x106/ml. Antibody solution was aspirated, and wells were washed with complete media followed by addition of 2 x106 cells to each well. Activation was carried out at 37° C.
Activated T cells were harvested, washed and counted. Transduction mixtures containing concentrated virus supernatants, protamine sulfate (10 μg/ml), IL-7 (10
ng/ml) and IL-15 (100 ng/ml) were prepared. For each transduction, 1x106 cells were separated in a sterile microcentrifuge tube and centrifuged at 400xg for 5 minutes. Each
cell pellet was resuspended in 0.5 ml of the transduction mixture corresponding to a specific MOI. Cell suspension was placed in an appropriately labelled well of a 24-well
G-Rex plate. After 24 hours of incubation at 37° C. and 5% CO2 , 1.5 ml media supplemented with IL-7 (10 ng/ml) and IL-15 (100 ng/ml) was added to each well. Ninety-six-hour post-transduction, transgene expression was determined by
flow cytometry. Multimeric MHC-peptide complexes (Dextramer or Tetramer) were used to monitor surface expression of transgenic TCR by FACS.
The method of Kalra achieved successful transduction with each of two different lentiviral constructs encoding R7Pl D5 TCR, i.e., LV-R73 and LV-R78 (Example 2 and Figure 14.
Kalra teaches that their TCR expressing T cells can be used to treat cancer by reducing the size of a tumor or preventing the growth or re-growth of a tumor in these subjects (paragraphs 0159-0169). As can be seen in Figure 23D, T cells expressing transduced TCR were incubated with tumor cells pulsed with different concentration of target peptide, followed by measuring tumor cell growth. FIG. 23D shows expanded T cells expressing transduced TCR inhibit tumor cell growth in a peptide concentration dependent manner (Example 4 and Figure 23D). As such, additional anticancer molecules (bioproducts with anticancer activity) are also released by the T cells after the TCR binds to antigen on the cancer cell.
Regarding claims 6 and 10, Kalra teaches that autologous TCR-Engineered T cells can be used to treat solid tumors (Example 6).
Regarding claim 15, Kalra teaches that the cancer being treated can be breast cancer (paragraph 0140).
Regarding claim 16, as stated in the 112b rejection above, there is a lack of clarity whether epidural and infusion are one route of administration or separate administrations. For the purposes of this rejection, epidural and infusion are considered separate routes of administration.
Kalra teaches that a continuous infusion can be used to deliver the T cells to a patient (paragraph 0169).
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.
Claims 1 and 11 are rejected under 35 U.S.C. 103 as being unpatentable over United States Patent Application No. 20190247433 (Kalra) as applied to claim 1 above, and further in view of Berger et al. (Expert Opinion on Biological Therapy 7: 1167-1182. 2007).
The teachings of Kalra are as discussed above.
Kalra does not teach wherein the patient also undergoes chemotherapy.
However, Berger teaches that they performed chemotherapy in melanoma patients prior to administering T cells expressing TCRs to the cancer patients. Berger teaches that chemotherapy depleted endogenous lymphocytes to promote the survival of the transferred T cells (page 1170, column 1, paragraph 1).
It would have been obvious to one of ordinary skill in the art before the effective filing date of the claimed invention to have combined chemotherapy with the method of treating cancer with T cells expressing TCRs of Kalra to arrive at the instantly claimed invention. One of ordinary skill in the art would have a reason to combine with a reasonable expectation of success because Berger teaches that performing chemotherapy prior to administering TCR expressing T cells depletes endogenous lymphocytes to promote the survival of the transferred T cells, increasing the efficacy of the treatment. Because the prior art teaches all of the elements of the claimed invention, there is a reasonable expectation of success.
Claims 1, 11-12, and 14 are rejected under 35 U.S.C. 103 as being unpatentable over United States Patent Application No. 20190247433 (Kalra) as applied to claims 1 and 11 above, and further in view of Yang et al. (Human Gene Therapy Methods 23: 73-83. 2012).
The teachings of Kalra are as discussed above.
Kalra does not teach wherein the treated cells are treated two times.
However, Yang teaches a method of transducing peripheral blood lymphocytes (including T cells) with a MART-1 TCR to target melanoma, comprising:
For lentiviral vector production, Vesicular stomatitis virus glycoprotein (VSV-G)-
pseudotyped lentiviral vector was produced transiently by transfection, using calcium phosphate and a four-plasmid system. The concentrated vector was treated with Benzonase (50 U/ml) for 1 hr at room temperature followed by diafiltration with 5 volumes (10 liters) of AIM-V medium. The final product was aliquoted into 50-ml Cryocyte bags and stored at - 80°C until further use.
For PBL transduction, On day 0, PBLs (5 x 106 cell/ml, 2 ml/well) were stimulated with interleukin (IL)-2 (300 IU/ml) and anti-CD3/anti-CD28 beads (ratio of beads to cells was 3:1) in tissue culture-treated 6-well plates and incubated overnight at 37°C and 5% CO2. Serial dilutions of vector supernatant containing protamine sulfate (10 lg/ml) were applied to 1 x 106 stimulated PBLs (final volume, 1 ml) in individual wells of a 24-well plate. The plates were centrifuged at 2000 · g for 2 hr at 32°C followed by incubation at 37°C and 5% CO2. The next day, the medium was replaced with AIM-V medium containing 5% human serum and IL-2 (300 IU/ml), and the cells were transferred to 25-cm2 tissue culture flasks at a density of 0.3 x 106 cells/ml.
The current World Health Organization (WHO) regulatory guidance limit for residual DNA in biological products derived from continuous cell lines is 10 ng/dose. The concentration of residual plasmid DNA in the vector supernatant after Benzonase treatment was below the detection limit of the assay (2.0 ng of DNA per milliliter;
Supplementary Table S1) whereas without the benzonase treatment, the residual DNA was ~12 ng/mL, above the guidance limit.
Yang identifies that treating the lentiviral supernatant with benzonase leads to residual benzonase during transduction of the PBLs and does not reduce PBL transduction efficiency or function, leading to expression of the TCR in the PBLs (Figure 3).
Yang teaches that their simplified vector production method can more easily meet the needs of small-scale phase I/II clinical trials for transduction of mature lymphocytes while complying with the current regulatory standards. This simplified production process will decrease the amount of time and effort required for process development, as well as, eliminate the need for concentration and diafiltration, thereby decreasing both the processing time and cost for production for a specific lentiviral vector product.
It would have been obvious to one of ordinary skill in the art before the effective filing date of the claimed invention to have modified the method of manufacturing and administering T cells expressing TCRs to treat cancer of Kalra by using the vector production method of Yang to arrive at the instantly claimed invention. One of ordinary skill in the art would have a reason to modify with a reasonable expectation of success because Yang teaches that their simplified vector production method can more easily meet the needs of small-scale phase I/II clinical trials for transduction of mature lymphocytes while complying with the current regulatory standards. This simplified production process will decrease the amount of time and effort required for process development, as well as, eliminate the need for concentration and diafiltration, thereby decreasing both the processing time and cost for production for a specific lentiviral vector product. Therefore, it would have been obvious to use the method of vector production of Yang as it is faster, cheaper, and easier. Because the prior art teaches all of the elements of the claimed invention, there is a reasonable expectation of success.
Using the combined method of producing T cells of Kalra and Yang would lead to two different benzonase treatments, one after thawing PBMCs (as taught by Kalra) and one during transduction of the cells (as taught by Yang).
Claims 1, 5, 10-11, 13, 16, and 20 are rejected under 35 U.S.C. 103 as being unpatentable over Vadhan-Raj et al. (The Lancet 359: 2145-2152. 2002), Federici et al. (Thrombosis Research 129: S60-S65. 2012), Chen et al. (BMC Immunology 21: 1-13. 2020), and Fry et al. (Thesis: Cell Separation and Cryopreservation of Cord Blood Fractions for Immunotherapeutic Applications. 2014).
Vadhan-Raj teaches that cryopreserved autologous and/or allogeneic platelets can be administered to cancer patients experiencing severe chemotherapy-induced thrombocytopenia through a transfusion (i.e. intravenous administration) and that platelets were collected by plateletpheresis (abstract and page 2146, column 2, paragraphs 2-4).
Vadhan-Raj is silent regarding whether leukocytes are also delivered and does not teach that they treated these cells with benzonase.
However, Federici teaches that platelets can be administered to cancer patients experiencing thrombocytopenia associated with neoplasia. As part of the platelet transfusion, platelets are collected through apheresis procedures (also known as plateletpheresis) which leads to a leukocyte-reduced (but not eliminated) platelet concentrate (abstract and page S62, column 1, paragraph 2- page S63, column 1, paragraph 1).
Chen teaches that benzonase can be added to isolated cryopreserved cells from the blood after they have thawed (page 11, column 1, paragraph 2). As can be seen in Figure 2, thawing cryopreserved cells leads to cell death, which leads to cell free DNA within the media after thawing.
Fry teaches that DNase (of which benzonase is known to have DNase activity) is often used to break down cell free DNA after thawing bone marrow cells (page 22, paragraph 2).
It would have been obvious to one of ordinary skill in the art before the effective filing date of the claimed invention to have modified the method of administering thawed cryopreserved leukocyte reduced platelets to cancer patients experiencing thrombocytopenia during chemotherapy treatment by also treating cancer patients experiencing thrombocytopenia associated with neoplasia, as identified by Federici, and treating the cells with benzonase after thawing to remove cell free DNA, as identified by Chen and Fry,to arrive at the instantly claimed invention. One of ordinary skill in the art would have a reason to modify with a reasonable expectation of success because Federici teaches that apheresis leads to leukocyte reduction but still retains some leukocytes and that leukocyte reduced platelets can be administered to cancer patients experiencing thrombocytopenia associated with neoplasia, Chen shows that thawing cryopreserved blood cells leads to cell death and successfully reduces to practice that benzonase can be added to blood cells after they have thawed, and Fry teaches that the benefit to using DNases, such as benzonase, after thawing cryopreserved cells is to remove cell free DNA. Therefore, it would have been obvious to treat the platelets/leukocytes with benzonase after thawing to remove any cell free DNA associated with cell death before administering the cells to treat thrombocytopenia associated with neoplasia and chemotherapy induced thrombocytopenia.
Conclusion
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/KEENAN A BATES/Examiner, Art Unit 1631