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 .
Continued Examination Under 37 CFR 1.114
A request for continued examination under 37 CFR 1.114, including the fee set forth in 37 CFR 1.17(e), was filed in this application after final rejection. Since this application is eligible for continued examination under 37 CFR 1.114, and the fee set forth in 37 CFR 1.17(e) has been timely paid, the finality of the previous Office action has been withdrawn pursuant to 37 CFR 1.114. Applicant's submission filed on May 08, 2026, has been entered.
DETAILED ACTION
Acknowledgement is hereby made of receipt and entry of the communication filed on May 08, 2026. Claims 1, 6-13, 16-19 and 25-32 are pending and are currently examined.
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.
Claims 19 and 25-30 and 32 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.
Claims 19 and 32 recite a phrase “…a clonal cell line derived from HEK293F cells…”, which renders the claims indefinite. It is unclear what “a clonal cell line” is represented for. The instant specification discloses that in some embodiments of the above method, the cells are a clonal cell line derived from HEK293F cells. In still other embodiments, the clonal cell line is Viral Production Cells 2.0 (VPC2.0) (See [0019]). So, it is not clear if there are other cell lines beside the VPC2.0.
In addition, the instant specification does not disclose 500% greater yields in “ a clonal cell line” compared with HEK293 cells except the VPC2.0. The instant specification discloses ”at least 500% greater” in VPC2.0 compared with HEK293F cells (See e.g., [0013], [0063]). Therefore, it is unclear what cell line is to be used to compare to the HEK293F for the “500% greater” in these claims.
Accordingly, one of ordinary skill in the art will not know the metes and bounds of the claims.
For purposes of compact prosecution and applying prior art, the “a clonal cell line” was interpreted to a cell line similar to the HEK293F cell and the claim 19-(3) and claim 32-(3) will not be addressed in the current office action because it is unclear what the cell line is used to be compared to HEK293F.
It is noted any interpretation of the claims set forth above does not relieve Applicant of the responsibility of responding to this rejection. If the actual interpretation of the claims is different than that posited by the Examiner, additional rejections and art may be readily applied in a subsequent final Office action.
Claims 31 and 32 recites “consists essentially of” that renders the claims indefinite because the instant specification fails to define the basic and novel properties of the invention.
For purposes of compact prosecution and applying prior art, the “consist essentially of” was interpreted herein as “comprise”. Also, see also MPEP § 2111.03.
It is noted any interpretation of the claims set forth above does not relieve Applicant of the responsibility of responding to this rejection. If the actual interpretation of the claims is different than that posited by the Examiner, additional rejections and art may be readily applied in a subsequent final Office action.
Claim Rejections - 35 USC § 103
The following is a quotation of 35 U.S.C. 103 which forms the basis for all obviousness rejections set forth in this Office action:
A patent for a claimed invention may not be obtained, notwithstanding that the claimed invention is not identically disclosed as set forth in section 102, if the differences between the claimed invention and the prior art are such that the claimed invention as a whole would have been obvious before the effective filing date of the claimed invention to a person having ordinary skill in the art to which the claimed invention pertains. Patentability shall not be negated by the manner in which the invention was made.
(New Rejection-necessitated by amendment) Claims 1, 6-13, 16-19 and 25-32 are rejected under 35 U.S.C. 103 as being unpatentable over Zhang et al.( WO2007059473A2, published on May 24, 2007, hereinafter, “zhang”) in view of Narayan et al. (Front Oncol. 2024 Mar 15;14:1359725), Labant (Gen, 2021, https://www.genengnews.com/topics/genome-editing/the-next-generation-of-cell-factories-for-viral-vector-production/) as evidenced by Yamada et al. (Cytotechnology. 2009 Apr;59(3):153-60).
The base claim 1 is directed to a method of manufacturing Nadofaragene firadenovec, the method comprising the steps of:
a) infecting Viral Production Cells 2.0 (VPC2.0 cells) grown in suspension with
Nadofaragene firadenovec; and
b) harvesting the Nadofaragene firadenovec produced by the VPC2.0 cells,
wherein the Nadofaragene firadenovec is produced in an amount at least 500% greater than a method in which HEK293 cells are infected with Nadofaragene firadenovec.
The base claim 19 is directed to a method of manufacturing Nadofaragene firadenovec, the method comprising the steps of:
a) infecting a clonal cell line derived from HEK293F cells grown in suspension with Nadofaragene firadenovec, wherein: (1) the target infection cell density is from about 1 x 10^6 to about 4 x 10^6 viable cells/mL; (2) infection is carried out at a multiplicity of infection of about 100 to about 300 and a duration of infection is about 48 hours;
and b) harvesting the Nadofaragene firadenovec produced by the clonal cell line.
Zhang teaches a method for manufacturing adenovirus compositions (See Abstract). The method disclosed by Zhang comprises the steps of (a) infecting (See [0164]) HEK293 cells grown in suspension (See [0111], [0112], [0113], [0118]) with an Ad5-based vector encoding human interferon (paragraph [0185], [0036]), and (b) harvest the adenovirus as “the cells may be harvested on day 1, 2, 3, 4, 5, 6 post infection. The virus yield can be up to 2.3 x 10^11 viral particles/mL or 230,000 viral particles/cell or more” (See [0025]), wherein the target infection cell density ranges from about 5 x 10^5 to about 2 x 10^6 cells/m L (from about 1 x 10^6 to about 4 x 10^6 viable cells/ml, as recited in the instant claim 19-a)-(1)) (see [0080]), and infection is carried out at an MOI range of 5-1000 (MOI of about 100 to about 300, as recited in claim 19-a)-(2) (See [0024]), and a duration of infection ranges from 1-10 or more days after viral infection (about 48 hours, as recited in claim 19-a)-(2)) (see [0027]); and (b) harvesting the Ad5 vector containing the transgene encoding human interferon produced by the HEK293 cells (see [0212], [0036]).
For both claims 1 and 19, they require the adenovirus is Nadofaragene firadenovec. Although Zhang does not explicitly discuss Nadofaragene firadenovec, Zhang teaches that the adenovirus is a replication-deficient adenovirus lacking at least a portion of the E1 region. In certain embodiments, the adenovirus may be lacking at least a portion of the E1 A and/or E1B region. In other embodiments, the adenovirus is a recombinant adenovirus that can be types 2 and 5 (Ad2 and Ad5, respectively) (See [0004] and [0110]), where the Ad5 vector contains and delivers the interferon α (See [0036]). Here the Ad5 virus, which is with E1 gene deletion and containing interferon α, is comparable to the Nadofaragene firadenovec.
The Nadofaragene firadenovec can be taught by Narayan’s study. Narayan teaches that Nadofaragene firadenovec-vncg received FDA approval as the first gene therapy in urology and is a non-replicating adenoviral vector expressing human IFNα2b (See page 3, right column, paragraph 1), which is from deletion of E1 and E3 genes from the human serotype 5 adenovirus (See page 3, left column, paragraph 1).
It would have been prima facie obvious for one having ordinary skill in the art before the effective filing date of the claimed invention to introduce the Nadofaragene firadenovec of Narayan into Zhang’s method to arrive at an invention as claimed. Zhang’s Ad5 vector contains comparable features as the Nadofaragene firadenovec of Narayan that includes the E1 deletion and encode an interferon α gene, and Zhang’s invention provides a high yield of adenoviral vector production (See [0012]). At the same time, Narayan teaches the Nadofaragene firadenovec is the first FDA-approved gene therapy for high-risk BCG-unresponsive NMIBC with CIS with or without papillary tumors (See page 5, left column). Thus, one of skill in the art would have been motivated to introduce the Nadofaragene firadenovec into Zhang’s invention to manufacture the Nadofaragene firadenovec with high yields for gene therapy application. There would be a reasonable expectation of success in producing the Nadofaragene firadenovec using Zhang’s method because a similar feature of Ad5 virus is successfully produced in the HK293F cell carrying DNA in either the E1, the E3 or both regions (See Zhang. [0112]).
Based on the description above, Zhang in view of Narayan teaches claims 1 and 19 for the Nadofaragene firadenovec limitation. At this point, Zhang teaches the limitations of a) and b) of claim 19.
As for the Viral Production Cells 2.0 (VPC2.0 cells) required in claim 1, Labant teaches that Thermo Fisher developed CTS Viral Production Cells (VPCs) for the production of scalable, high-titer lentivirus in suspension HEK293 cells. Three of these lines—the company’s FreeStyle 293-F cells, Viral Production Cells (VPCs), and VPCs 2.0—have been evaluated for their ability to produce adeno-associated virus (AAV) in suspension culture using the Gibco AAV-MAX Helper-Free AAV Production System. VPCs 2.0, a clonal 293 cell line selected for high AAV expression, generated AAV titers 45-fold and 10-fold higher than those generated by FreeStyle 293-F cells and VPCs, respectively (See page 3, and Figure below).
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Here the AAV yield in VPC2.0 is 45x compared to 293FS cell. Although adenovirus and AAV are different viruses, it is reasonable to consider that the yields for adenovirus should be also increased in the VPC2.0 cells and the 500% increase (which is about 6 times (X) greater compared to the HEK293 cell) as claimed should be achieved through routine experimental optimization. The following teachings can be the explanations:
1). The VPC 2.0 is a clonal HEK293 cell line (See Labant, page 2). Here the 293 cell line is a permanent line established from primary embryonal human kidney transformed with sheared human adenovirus type 5 DNA (Graham et al., 1977; Harrison et al., 1977). The E1A adenovirus gene is expressed in these cells and participates in transactivation of some viral promoters, allowing these cells to produce very high levels of protein (See: https://www.thermofisher.com/us/en/home/references/protocols/cell-culture/transfection-protocol/freestyle-293-expression-system.html), which indicates that the VPC2.0 and HEK293 can be infected by non-replication adenovirus such as the non-replication Ad5 of Zhang and Nadofaragene firadenovec of Narayan because the HEK293 cells and VPC.20 both contain the adenoviral E1 gene. This can be further evidenced by Yamada’s study. Yamada teaches that the replication-deficient, first-generation adenovirus vectors are based on substitution of their essential E1 genes by the therapeutic gene to be delivered to the target cells. The production of replication-deficient, E1-deleted adenovirus vectors is achieved by infecting a complementary cell line, such as the human embryonic kidney 293 cell line (Graham et al. 1977), which constitutively expresses E1 genes to support the replication of E1-deficient adenoviral particles (See page 154, left column, paragraph 2).
2). At the same time, Zhang teaches that the generation and propagation of the adenoviral vectors depending on a unique helper cell line, designated 293, which was transformed from human embryonic kidney cells by Adenovirus serotype 5 (Ad5) DNA fragments and constitutively expresses E1 proteins (Graham et al., 1977). Since the E3 region is dispensable from the Ad genome (Jones and Shenk, 1978), the current Ad vectors, with the help of 293 cells, carry foreign DNA in either the E1, the E3 or both regions (See [0112]).
Accordingly, Zhang teaches a 293 cell line that contains the same features such as E1 gene of Ad5 as the VPC2.0 of Labant. At the same time, Zhang also teaches that the 293 cell line can be used to infect with a replication-deficient adenovirus Ad5 vector lacking at least a portion of the E1 region and contain interferon α (See [0110], [0036]), wherein this recombinant Ad5 vector of Zhang is equivalent to the Nadofaragene firadenovec of Narayan.
It would have been prima facie obvious for one having ordinary skill in the art before the effective filing date of the claimed invention to introduce the Nadofaragene firadenovec of Narayan and the VPC2.0 cells of Labant into Zhang’s method to arrive at an invention as claimed. Zhang teaches that the Ad5 vector contains comparable features like the Nadofaragene firadenovec of Narayan that has the E1 deletion and encode an interferon α gene, which can be produced from a 293-cell line that was transformed from human embryonic kidney cells constitutively expressing E1 proteins, which is similar to VPC2.0. At the same time, Narayan teaches the gene therapy application of Nadofaragene firadenovec, and Labant teaches that the VPC2.0 is a high viral yield cell line based on the production of AAV. Thus, one of skill in the art would have been motivated to introduce Nadofaragene firadenovec of Narayan and the VPC2.0. of Labant into Zhang’s invention to test the yields. There would be a reasonable expectation of success to produce a high yield such as 500% increase of Nadofaragene firadenovec in the VPC2.0 cell line based on the teaching from the rationality taught by Zhang and the method used to produce a 45 x higher yields of the AAV in the VPS2.0 cells taught by Labant.
Thus, the invention as a whole was clearly prima facie obvious to one of ordinary skill in the art before the effective filing date of the claimed invention.
Regarding claims 6, 7, 8 and 25, Zhang teaches isolating the adenovirus from the adenovirus preparation at least about, at most about, or about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more days after viral infection is completed (teaches harvested at least 24 hours after infection, as recited in claim 6; harvested at least 36 hours after infection, as recited in claim 7; harvested about 48 hours after infection, as recited in claim 8) (see [0027]), where the “at least 24 hours, 36 hours or 48 hours” also teaches claim 25.
Regarding claims 9 and 26, Zhang teaches that Viral Production Cells (VPCs), and VPCs 2.0—have been evaluated for their ability to produce adeno-associated virus (AAV) in suspension culture using the Gibco AAV-MAX Helper-Free AAV Production System (See Labant, page 2, figure). Zhang also teaches that the host cell is adapted for growth in suspension culture. In particular embodiments, the cells of the present invention are designated IT293SF cells (See [0018]).
Regarding claims 10-11 and 27, Zhang teaches infection of the cells at a multiplicity of infection (MOI) of the vector of at least about, at most about, or about 5…100, 200, 300 …700 to about 500…900, 1000 MOI (MOI is about 100 to about 300, as recited in claim 10; MOI is about 175 to about 200, as recited in claims 11 and 27) (See Zhang, [0024]).
Regarding claim 12, 13 and 28, as discussed above, Zhang teaches the cell culture in a bioreactor, wherein the cells are the 293 derived cell lines which are found to be equivalent to VPC2.0 cells as discussed above, and are at a cell seeding density of about 7.5 x 10^5 cells/ml ( teaches claims 12 and 28) (See Zhang, [0080]), and the infection cell density can be 2x10^6 (See Zhang, [0283]) (teaches claim 13). Zhang also teaches using advanced probes and autoanalyzers for on-line and off-line measurements of turbidity (a function of particles present), capacitance (a function of viable cells present) (See [0137]), which indicates that the cell density for seeding and infection are all monitored by its viability.
Regarding claims 16-18 and 29-30, Zhang further teaches harvesting virus supernatant after complete cell lysis, wherein the lysing step can be accomplished with a wide variety of agents (See Zhang, [0212], Table 6) (lysing the producer cells with a lysis agent prior to harvesting, as recited in claims 16 and 29). Zhang in view of Labant teaches a step of density gradient centrifugation of the lysed cells (See Zhang, [0257]) (centrifuging the lysed cells, as recited in claim 17). Zhang also teaches purifying the viral vector after harvesting, to a pharmaceutically acceptable degree (See Zhang, [0031]) (purifying the vector after harvesting, as recited in claims 18 and 30).
Regarding claim 31, based on the description above, Zhang teaches growing a 293 cell, which contains Ad5 E1 gene that is equivalent to the APC2.0 cells of Labant, and infecting the cells with a Ad5 adenovirus lacking at least a portion of the E1 region and carrying the interferon α gene, that is equivalent to the Nadofaragene firadenovec of Narayan and harvest at day 1, 2, 3, 4, 5, 6 post infection (See [0025]). Here the description teaches the claim 31-a) and 31-b).
As for the “at least 500% greater than a method in which HEK293 cells are infected with Nadofaragene firadenovec”, Zhang teaches an equivalent cell line as VPC2.0 and teaches a Ad5 vector that is equivalent to Nadofaragene firadenovec as well. Zhang discloses that the virus yields in 200L bioreactor would be expected to yield approaching 2 x 10^16 vp or more (See [0025]) and the virus yields can be further increased by optimizing the seeding/infecting cell density (See [0080], [0087]), culture temperature (See [0081]), MOI (See [0086]) and media supplantation ([0088]). Therefore, one of skill in the art can achieve a 500% or 6x greater yield by infecting the VPC2.0 with the Nadofaragene firadenovec through a routine experimental optimization based on the condition disclosed by Zhang.
Regarding claim 32, based on the description above, Zhang teaches growing a 293-cell containing Ad5 E1 gene and infecting with a Ad5 adenovirus lacking at least a portion of the E1 region and carrying the interferon α gene, which is equivalent to the Nadofaragene firadenovec. Zhang teacehs the infection cell density can be 2x10^6 and the multiplicity of infection (MOI) of the vector of at least about 5…100, 200, 300 …700, and harvest at is two days post infection (48hr) (See [Zhang, [0345]). Thus, Zhang teaches claims 32- a) and b).
.
Responses to Applicant’s Remarks
Applicant’s arguments filed on May 08, 2025, has been received and fully considered.
1. Applicant’s argument on Zhang in view of Labant is not found persuasive (See Remarks, pages 6-9).
1). Labant teaches generating a VPC2.0 cell line and discloses that it is derived from a clonal HEK293 cell line, which contains the Ad5 E1 gene. Therefore, the Nadofaragene firadenovec, an Ad5 vector lacking E1 gene, can express in the VPC2.0 cells.
2). Zhang already teaches a 293-cell line that is comparable to the VPC2.0 cells (See Zhang, [0112]), which can be infected by an Ad5 vector containing interferon α gene and lacking of E1 gene. Here the Ad5 is comparable to the claimed Nadofaragene firadenovec, Zhang also discloses that the virus yields in 200L bioreactor would be expected to approach to 2 x 10^16 vp or more (See [0025]) and the virus yields can be further increased by optimizing the seeding/infecting cell density (See [0080], [0087]), culture temperature (See [0081]), MOI (See [0086]), media supplantation ([0088]), etc. Therefore, one of skill in the art can develop a method for achieving a 500% or 6x greater yield of Nadofaragene firadenovec in VPC2.0 through routine experimental optimizations.
3). On the other hand, Labant’s VPC 2.0 is a clonal HEK293 cell (See Labant, page 2), so, Nadofaragene firadenovec or Zhang’s Ad5 virus can infect and grow in the VPC2.0 cell. It is a fact that the AAV and Adenovirus is not the same, however, the 45-fold higher yields of AAV in the VPCs 2.0 compared to the HEK293F (See below) would provide a motivation for one of skill in the art to set up an experiment to test the production yields on other viruses such as Adenovirus/ Nadofaragene firadenovec. Zhang teaches that “Clearly, it becomes unfeasible to produce the required amount of virus using this traditional process. New production and purification processes that can be scaled up and validated have to be developed to meet the increasing demand” (See [0007]).
This is also a response to applicant’s argument at “The Examiner's Articulated Motivation to Combine Does Not Support Obviousness Under § 103” (See Remarks, page 11).
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Note: the references of Shiina, Tan, Hassinen are not applied in the current office action. The arguments regarding these references are moot.
2. Applicant’s argument on the unexpected result is not found persuasive (See Remarks, pages 7 and 12-14).
Applicant argued the unexpected result by listing Example 1 in the instant specification. The alleged unexpected result is not persuasive because the 570% greater yield in VPC 2.0 compared to HEK293 is in specific conditions at 4x2L bioreactor, pH 7.2 and a specific agitation rpm with specific harvest time. However, the base claims 1 and 31 are directed to a generic method for producing Nadofaragene firadenovec in the VPC2.0 cell “at least 500% greater than a method in which HEK293 cells”.
Therefore, applicant’s alleged unexpected results are not commensurate with the scope of the claims.
3. Applicant argued the newly added claims 31 and 32 for the term "consists essentially of” and the prior art reference (See Remarks, page 14).
The argument is not persuasive.
1). The reference Shiina is not applied in the current office action. Thus, the argument regarding Shiina is moot.
2). See also MPEP § 2111.03. A ‘consisting essentially of’ claim occupies a middle ground between closed claims that are written in a ‘consisting of’ format and fully open claims that are drafted in a ‘comprising’ format." PPG Indus. v. Guardian Indus., 156 F.3d 1351, 1354, 48 USPQ2d 1351, 1353-54 (Fed. Cir. 1998). For the purposes of searching for and applying prior art under 35 U.S.C. 102 and 103, absent a clear indication in the specification or claims of what the basic and novel characteristics actually are, "consisting essentially of" will be construed as equivalent to "comprising."
4. Applicant argued the Dependent Claims 6-13, 16-18, and 25-30 are patentable because of they were cited with additional limitation (See the Remarks, page 15).
The argument is not persuasive.
The limitations cited to the claims 6-13, 16-18, and 25-30 are taught by Zhang in view of the supported prior art references. Thus, the 103 rejection is applicable to the claims.
Conclusion
No claims are allowed.
Any inquiry concerning this communication or earlier communications from the examiner should be directed to RUIXUE WANG whose telephone number is (571)272-7960. The examiner can normally be reached Monday-Friday 8:00 am..
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/RUIXUE WANG/Examiner, Art Unit 1672
