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 .
DETAILED ACTION
Priority
The present application claims the priorities from the U.S. provisional patent application serial number 63/593,242 filed October 26, 2023, that is hereby acknowledged by the Examiner.
Status of the Claims
The amendment dated 10/18/2024 is acknowledged. Claims 1-17 are pending and under examination.
Information Disclosure Statement
The information disclosure statement (IDS) submitted on 11/14/2024 and 06/22/2026 are in compliance with the provisions of 37 CFR 1.97. Accordingly, the information disclosure statement(s) is/are being considered by the Examiner.
Drawings
The drawing filed on 10/18/2024 are acknowledged and accepted by the Examiner.
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.
Claims 12-13 are rejected under 35 U.S.C. 112(b), as being indefinite for failing to particularly point out and distinctly claim the subject matter which applicant regards as the invention.
Claims 12 and 13 recite “the step of measuring impact of the therapeutic compound”. There is insufficient antecedent basis for this limitation in the claim. Accordingly, one of ordinary skill in the art would not know the metes and bounds of the claims.
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 35 U.S.C. 112 (pre-AIA ), fourth paragraph:
Subject to the [fifth paragraph of 35 U.S.C. 112 (pre-AIA )], 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 10 is rejected under 35 U.S.C. 112(d) or 35 U.S.C. 112 (pre-AIA ), 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 10 depends from claim 7, whereby claim 7 already limits the method to primary human alveolar epithelial cells that are isolated from an non-malignant lung tissue. 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 § 103
The following is a quotation of 35 U.S.C. 103(a) which forms the basis for all obviousness rejections set forth in this Office action:
(a) A patent may not be obtained though the invention is not identically disclosed or described as set forth in section 102 of this title, if the differences between the subject matter sought to be patented and the prior art are such that the subject matter as a whole would have been obvious at the time the invention was made to a person having ordinary skill in the art to which said subject matter pertains. Patentability shall not be negatived 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 under 35 U.S.C. 103(a), the examiner presumes that the subject matter of the various claims was commonly owned at the time any inventions covered therein were made absent any evidence to the contrary. Applicant is advised of the obligation under 37 CFR 1.56 to point out the inventor and invention dates of each claim that was not commonly owned at the time a later invention was made in order for the examiner to consider the applicability of 35 U.S.C. 103(c) and potential 35 U.S.C. 102(e), (f) or (g) prior art under 35 U.S.C. 103(a).
Claims 1-17 are rejected under 35 U.S.C. 103(a) as being unpatentable over Chan et al. “Chan” (PNAS, March 29, 2016, 113(13):3621-3626, IDS of record dated 11/14/2024; and supporting information 2016, 113(13):1-5) in view of Saxena (ed.), High-throughput Screening for Drug Discovery, May 25, 2022, ISBN: 978-1-83962-948-8, Chapter 4, IDS of record dated 06/22/2026).
The claims are directed to a high-throughput in vitro lung injury model system for screening a plurality of candidate compounds for the treatment of respiratory infectious diseases, comprising:
a plurality of transwell inserts in a 96-well system, wherein each transwell insert contains a monolayer of primary human alveolar epithelial cells, and the primary human alveolar epithelial cells are infected with one or more respiratory pathogens; a device for applying a plurality of candidate compounds to infected alveolar epithelial cells; and an apparatus for measuring alveolar fluid clearance and alveolar protein permeability in the infected alveolar epithelial cells.
Regarding claim 1, Chan discloses an in vitro model of acute lung injury for studying the effect of mesenchymal stromal cells (MSCs) on alveolar fluid clearance and protein permeability impaired by influenza H5N1 virus infection in human alveolar epithelial cells (page 3625, first column, first para.; pages 3626, first column, second para. to sixth para.; Fig. 1 and 2; and supporting information, page 1 second column, second para. to page 2, first column, first para.). Chan discloses that alveolar epithelial cells were plated on the apical chamber of 24-well Costar Transwell inserts with a 0.4 um pore size at a density of 100,000 cells per well; the microporous transwell membrane established an air-liquid interface similar to that of human lung epithelium, and the plated cells were maintained in humidified atmosphere (5% CO2, 37oC); for coculture experiments with alveolar epithelial cells, MSCs or normal adult human lung fibroblasts (controls) were plated on the basolateral chamber of the transwell (Fig. 1) at a density of 100,000 cells per well (or with different MSC cell density; 1,000, 10,000, 50,000 and 500,000 MSCs) and had no direct contact with the apical chamber (corresponding to a device for applying a plurality of candidate compounds to infected alveolar epithelial cells (supporting information, page 1, second column second para.); net alveolar fluid transport from the apical chamber of the transwell culture insert (containing a monolayer of alveolar epithelial cells infected with influenza virus) to the basolateral chamber of the transwell at 24 h p.i (corresponding to each transwell insert contains a monolayer of primary human alveolar epithelial cells, and the primary human alveolar epithelial cells are infected with one respiratory pathogen ( supporting information, page 1, second column, third para.); transepithelial resistance was maintained at 1,500 ohms/cm2, and there was morphological evidence of tight junctions; alveolar epithelial cells were either inoculated with the respective influenza A viruses at an MOI of 0.1 or incubated with the virus-free conditioned medium for 1 hour; then 200 ul of FITC-labeled dextran with a size of 70 kDa was added to the cells (final concentration, 500 ng/ul); after 5 min, the initial sample of 20 ul was collected from the apical chamber; after 24 h of dextran incubation, another 20 ul was collected from the same compartment, and 100 ul was collected from the basolateral chamber as the final samples; the fluorescence of each sample was measured by a modulus fluorometer (supporting information, page 1, second column, third para.); the net alveolar fluid transport in ul-cm-2-h-1 was measure as [1– (fluorescence reading in the initial sample/weight of the initial sample)/(fluorescence reading in the final sample/weight of the final sample)] × 100. The remaining fluid in the basolateral chamber was collected after 24 h of dextran incubation to measure protein permeability, based on the unidirectional flux of fluorescence-labeled dextran from the apical to the basolateral chamber of the transwell culture insert (corresponding to an apparatus for measuring alveolar fluid clearance and alveolar protein permeability I the infected alveolar epithelial cells) (supporting information, page 2, first column, first para.). Chan discloses that influenza A H5N1 virus-infected alveolar epithelial cells impair alveolar fluid clearance and protein permeability by down-regulating alveolar sodium and chloride transporter proteins; this pathology was prevented or reduced by coculture with MSCs in vitro and by MSC treatment in vivo in aged mice
(page 3625, first column, 1st paragraph). Chan provides the teaching of using an in vitro lung injury model to screen the therapeutic effect of MSCs treatment against the respiratory infectious disease caused by influenza A H5N1 virus by measuring alveolar fluid clearance and alveolar protein permeability in
infected alveolar epithelial cells.
Chan does not explicitly teach a 96-well plate and the cells are infected with more than one respiratory pathogen with a plurality of compounds applied to the cells.
Providing a high-throughput screening assay in order to improve the efficiency of screening multiple candidate compounds is obvious to one of ordinary skill in the art. A 96-well format is commonly used for high-throughput screening assays (see for example, Saxena section 1.3 “In screening”). Accordingly, it would have been obvious to one of ordinary skill in the art to adopt the 96-well format and to apply a plurality of candidate compounds to the 96-well system. One of ordinary skill in the art would have been motivated to do so with a reasonable expectation of success in order to improve the efficiency of the in vitro lung injury model of Chan. Moreover, it would be readily conceivable to one of ordinary skill in the art to infect the alveolar epithelial cells with more than one respiratory pathogens in the system in order to provide a high-throughput and efficient screening assay.
Regarding claims 2 and 3, Chan discloses that influenza H5N1 virus was used to infect the alveolar epithelial cells (supporting information, page 1, second column, second para. and Fig. S3). Additionally, respiratory syncytial virus (RSV), coronavirus, middle east respiratory syndrome coronavirus (MERS-CoV), human parainfluenza viruses (HPIVs), human metapneumovirus (hMPV), adenoviruses, rhinoviruses, and enteroviruses are common respiratory pathogens that cause respiratory infectious diseases.
Regarding claim 4, Chan discloses that alveolar epithelial cells were isolated from resected, nonmalignant lung tissues (page 3626, first column, fifth para.).
Regarding claim 5, Chan discloses that the alveolar epithelial cells were plated on the apical chamber of 24-well Costar Transwell inserts with a 0.4 um pore size, at a density of 100,000 cells per well (supporting information, page 1, second column, second para.).
Regarding claim 6, Chan discloses the effect of MSCs in reducing influenza A H5N1 acute lung injury (page 3625, second column, sixth paragraph to page 3626, first column, first paragraph
And supporting information, Fig. S3).
Regarding claim 7, the assessing effect of the plurality of candidate compounds on alveolar fluid clearance (AFC) and alveolar protein permeability (APP), Chan discloses Highly pathogenic influenza H5N1 virus significantly reduces human alveolar fluid clearance (AFC) and significantly increases human alveolar protein permeability (APP) compared with low pathogenic seasonal influenza viruses (supporting information Fig. S2).
Regarding claims 8-11, Chan discloses that influenza H5N1 virus was used to infect the alveolar epithelial cells (supporting information, page 1, second column, second para. and Fig. S3). Additionally, respiratory syncytial virus (RSV), coronavirus, middle east respiratory syndrome coronavirus (MERS-CoV), human parainfluenza viruses (HPIVs), human metapneumovirus (hMPV), adenoviruses, rhinoviruses, and enteroviruses are common respiratory pathogens that cause respiratory infectious diseases. Chan discloses that alveolar epithelial cells were isolated from resected, nonmalignant lung tissues (page 3626, first column, fifth para.). Chan discloses the effect of MSCs in reducing influenza A H5N1 acute lung injury (page 3625, second column, sixth paragraph to page 3626, first column, first paragraph and supporting information, Fig. S3). It is not inventive and considered routine and obvious to one of ordinary skill in the art to use MSCs as taught by Chan whereby the MSCs are nano-sized extracellular vesicles. One of ordinary skill in the art would be motivated to do so with a reasonable expectation of success given the knowledge that Chan discloses the use of MSCs; and utilizing nano-sized vesicles in order to provide a high-throughput and efficient screening assay.
Regarding claims 12-13, Chan discloses that the mRNA expression of cytokines (IFN-β, and IL-1β) and chemokines (RANTES, IP-10, MCP-1, IL6, and IL8) and of sodium and chloride transporters (CFTR; α-, β-, and γ-ENaC; α 1-, α 2-, α 3-, β1-, β2-, and β3-NaKATPase) by the infected cells was evaluated by using the SYBR Premix Ex Taq II kit (Takara) according to the manufacturer's instructions (supporting information, page 2, first column, second para.).
Regarding claim 14, a 96-well format is commonly used for high-throughput screening assays (see for example, Saxena section 1.3 “In screening”). Accordingly, it would have been obvious to one of ordinary skill in the art to adopt the 96-well format and to apply a plurality of candidate compounds to the 96-well system. In addition, liquid-liquid interface is a commonly used configuration for transwell system for cell study. Absent unexpected results to utilize a liquid-liquid interface, a person skilled in the art would readily adopt a liquid-liquid interface 96 transwell system based on the teachings of Chan in view of Saxena.
Regarding claim 15, Chan discloses the cells are seeded at a density of 1 x 104 to 1 x 105 cells per well (supporting information, second column, second para.).
Regarding claim 16, Chan discloses that alveolar epithelial cells were inoculated with the respective influenza A viruses at an MOI of 0.1 for the measurement of AFC and APP (supporting information, page 1, second column, third paragraph); and alveolar epithelial cells were inoculated with influenza viruses at an MOI of 2 for the measurement of cytokine responses of influenza virus-infected alveolar epithelial cells (supporting information, page 2, first column, second paragraph).
Regarding claim 17, formulating suitable concentrations of compounds for screening assays is a
conventional practice in the art. It is not inventive and considered routine and obvious to one of ordinary skill in the art to formulate suitable concentrations through routine experimental optimization. It has long been settled to be no more than routine experimentation for one of ordinary skill in the art to discover an optimum value of a result effective variable. According to section 2144.05 of the M.P.E.P., "[W]here the general conditions of a claim are disclosed in the prior art, it is not inventive to discover the optimum of workable ranges by routine experimentation." Application of Aller, 220 F.2d 454, 456, 105 USPQ 233, 235-236 (C.C.P.A. 1955). "No invention is involved in discovering optimum ranges of a process by routine experimentation." Id. at 458, 105 USPQ at 236-237. The "discovery of an optimum value of a result effective variable in a known process is ordinarily within the skill of the art." Application of Boesch, 617 F.2d 272, 276, 205 USPQ 215, 218-219 (C.C.P.A. 1980). It would have been obvious for one of ordinary skill to determine the appropriate concentrations of the compounds in the methods disclosed by the prior art by routine experimentation procedures known in the art. One of ordinary skill in the art would have been motivated to do so with a reasonable expectation of success in order to achieve the maximum (production or activity or therapeutic) response from the composition.
Therefore, the claimed invention would have been prima facie obvious to one of ordinary skill in the art before the effective filing date of the claimed invention.
Conclusion
No claims are allowed.
Any inquiry concerning this communication or earlier communications from the examiner should be directed to Barry Chestnut whose telephone number is (571)270-3546. The examiner can normally be reached on M-Th 8:00 to 4:00.
If attempts to reach the examiner by telephone are unsuccessful, the examiner’s supervisor, Thomas Visone can be reached on 571-270-0684. The fax phone number for the organization where this application or proceeding is assigned is 571-273-8300.
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/BARRY A CHESTNUT/Primary Examiner, Art Unit 1672
