Researchers Submit Patent Application, “Cell Culture Additives And Their Use For Increased Bioprotein Production From Cells”, for Approval (USPTO 20190085060): American Air Liquide Inc. | MarketScreener

Researchers Submit Patent Application, “Cell Culture Additives And Their Use For Increased Bioprotein Production From Cells”, for Approval (USPTO 20190085060): American Air Liquide Inc. | MarketScreener

May 08, 2019 0 Comments

2019 APR 05 (NewsRx) -- By a News Reporter-Staff News Editor at Health Policy and Law Daily -- From Washington, D.C., NewsRx journalists report that a patent application by the inventors LEONARDI, Jennifer (Wilmington, DE); SCHWARZ, Flavio (Wilmington, DE); CHIANG, Barbara (Wilmington, DE); TSENG, Alice (Wilmington, DE), filed on September 19, 2017, was made available online on March 21, 2019. The patent’s assignee is American Air Liquide Inc. (Houston, Texas, United States). News editors obtained the following quote from the background information supplied by the inventors: “Recombinant human insulin became the first authorized biologic in 1982. Junod, Suzanne White. ‘Celebrating a milestone: FDA’s approval of first genetically-engineered product.’ Update (2007): 43-44. The biologics industry has expanded dramatically since then with the addition of numerous new biologics, providing much needed therapeutic options for numerous diseases from hemophilia to cancer. In 2015, the U.S. FDA approved thirteen new biologics, nine of which were classified as ‘first-in-class’ therapeutics. FDA Center for Drug Evaluation and Research, ‘Novel Drugs 2015 Summary’, January 2016. . “The great promise of biologics as therapeutics faces numerous pragmatic challenges, not the least of which is the cost and complexity of production. The regulatory framework is also necessarily different than traditional small molecule pharmaceuticals: “’Because, in many cases, there is limited ability to identify the identity of the clinically active component(s) of a complex biological product, such products are often defined by their manufacturing processes. Changes in the manufacturing process, equipment or facilities could result in changes in the biological product itself and sometimes require additional clinical studies to demonstrate the product’s safety, identity, purity and potency. Traditional drug products usually consist of pure chemical substances that are easily analyzed after manufacture. Since there is a significant difference in how biological products are made, the production is monitored by the agency from the early stages to make sure the final product turns out as expected.’ “U.S. FDA, Frequently Asked Questions About Therapeutic Biological Products, . “Production and supply of biologics is therefore a key practical bottleneck in the flow of biologics to patients in need. Prices for biologics are correspondingly high relative to traditional small molecule chemical drugs. As a result, cost benefit analysis of biologics becomes critical in making decisions regarding access. See, e.g., Joensuu, Jaana T., et al. ‘The cost-effectiveness of biologics for the treatment of rheumatoid arthritis: a systematic review.’ PloS one 10.3 (2015): e0119683; Huoponen, Saara, and Marja Blom. ‘A Systematic Review of the Cost-Effectiveness of Biologics for the Treatment of Inflammatory Bowel Diseases.’ PloS one 10.12 (2015): e0145087. “In an effort to incentivize additional supply and lower costs, The U.S. federal government enacted the Biologics Price Competition and Innovation Act of 2009 as part of the Patient Protection and Affordable Care Act of 2010. This new law established the legal pathway and mandate for ‘biosimilars’ i.e. generic biologics to secure market authorization. Ahmed, Isiah, Ben Kaspar, and Uma Sharma. ‘Biosimilars: impact of biologic product life cycle and European experience on the regulatory trajectory in the United States.’ Clinical therapeutics 34.2 (2012): 400-419. The European Medicines Agency has been at the global forefront of biosimilars regulation with the first biosimilar approvals. Ibid. “One means of improving supply and reducing cost would be improved productivity through cost effective measures to increase product yield from existing biologics manufacturing. Ideally this would be accomplished with minimal capital expense. And of course, the resultant product would have to survive a rigorous regulatory review and evaluation analogous to proposed biosimilars or ‘major’ production process changes under FDA or other Medicines agencies’ regulations (e.g. EMA). “Due to the intense commercial and healthcare pressures on biologics production and cost, the state of the art for biopharmaceutical production involves extensive, multifaceted optimization and manipulation of many parameters in creating and culturing cells for protein biomolecule production. Almo S C, Love J D. Better and faster: improvements and optimization for mammalian recombinant protein production. Current opinion in structural biology. 2014; 26:39-43. doi:10.1016/j.sbi.2014.03.006; Mammalian Cell Cultures for Biologics Manufacturing (2014) Advances in Biochemical Engineering/Biotechnology 139 (Zhou, W. and Kantardjieff, A. eds). “Many cell culture process manipulations are routinely used. Abu-Absi, Susan, et al. ‘Cell culture process operations for recombinant protein production.’ Mammalian cell cultures for biologics manufacturing. Springer Berlin Heidelberg, 2013. 35-68. For example, culture parameters, chemical agents and genetic engineering approaches have all been applied to cause cell cycle inhibition to increase specific cellular productivity to boost protein biomolecule yields. Du, Zhimei, et al, ‘Use of a small molecule cell cycle inhibitor to control cell growth and improve specific productivity and product quality of recombinant proteins in CHO cell cultures.’ Biotechnology and bioengineering 112.1 (2015): 141-155. “Despite the extensive and ongoing efforts to improve protein biomolecule production, there remains intense commercial pressure and medical need to find new ways for further improvement.” As a supplement to the background information on this patent application, NewsRx correspondents also obtained the inventors’ summary information for this patent application: “The invention may be understood in relation to the following non-limiting, exemplary embodiments: “In a first embodiment, the invention may be described in part by the following numbered sentences: Sentence 1. A method of increasing protein biomolecule production comprising the steps of: a) culturing a cell that produces a protein biomolecule, b) supplying an Additive, preferably Nitric Oxide, and/or a source of the Additive to the culture medium in an amount sufficient to (i) increase a total yield of the protein biomolecule secreted into the cell culture media and/or (ii) increase a specific cellular productivity of the protein biomolecule secreted into the cell culture media, c) wherein the increase of step b) is measured relative to: culturing under a prior set of conditions established for a regulatory approval for producing the protein biomolecule and/or culturing under substantially the same conditions except for supplying the Additive and/or a source of the Additive. “Sentence 2. The method of sentence 1, wherein (a) the Additive is Nitric Oxide and/or a chemical source of Nitric Oxide, (b) the cell is a eukaryotic cell, and © the protein biomolecule is a heterologous protein biomolecule. “Sentence 3. The method of sentence 1 and/or 2, wherein the eukaryotic cell is a CHO cell and the heterologous protein biomolecule is a monoclonal antibody. “Sentence 4. The method of sentence 1, 2, and/or 3, wherein the Nitric Oxide is gaseous and is substituted for a portion of a volume of another gas being fed into the bioreactor. “Sentence 5. The method of sentence 4, wherein the other gas being fed into the bioreactor includes Oxygen gas, Nitrogen gas and/or or Carbon Dioxide gas. “Sentence 6. The method of sentence 1, 2, 3, 4, and/or 5, wherein the Nitric Oxide and/or the chemical source of Nitric Oxide are added in an amount sufficient to increase a total yield of the monoclonal antibody secreted into the cell culture media. “Sentence 7. The method of sentence 1, 2, 3, 4, 5, and/or 6, wherein the Nitric Oxide and/or the chemical source of Nitric Oxide are added in an amount sufficient to increase a specific cellular productivity of the monoclonal antibody secreted into the cell culture media. “Sentence 8. The method of sentence 1, 2, 3, 4, 5, 6, and/or 7, wherein the Nitric Oxide soluble in the cell culture medium is at a concentration of 0.1-10 nanomolar. “Sentence 9. The method of sentence 8, wherein the Nitric Oxide is supplied to the cell culture medium as a sparged gas at a concentration of 200-400 ppm Nitric Oxide in the sparged gas. “Sentence 10. The method of sentence 1, 2, 3, 4, 5, 6, 7, 8, and/or 9, wherein the eukaryotic cell is a CHO cell and the heterologous protein biomolecule is a monoclonal antibody. “Sentence 11. The method of sentence 1, wherein the Additive is Carbon Monoxide, (b) the cell is a eukaryotic cell, and © the protein biomolecule is a heterologous protein biomolecule. “Sentence 12. The method of sentence 11, wherein the eukaryotic cell is a CHO cell and the heterologous protein biomolecule is a peptide or glycopeptide biologic. “Sentence 13. The method of sentence 11 and/or 12, wherein the Carbon Monoxide is added in an amount sufficient to increase a specific cellular productivity of the monoclonal antibody secreted into the cell culture media. “Sentence 14. The method of sentence 13, further comprising the steps of supplying to the culture medium at least two additional Additives capable of inducing HO-1 activity and inhibiting caspase-3 activity in the cell. “Sentence 15. The method of sentence 14, wherein the Additive to induce HO-1 activity is selected from one or more of curcumin, caffeic acid phenethyl ester, resveratrol, quercetin, epigallocatechin gallate, carnosol, sulforaphane, and dimethyl fumarate. “Sentence 16. The method of sentence 14, wherein the Additive to inhibit caspase-3 activity is selected from one or more of quinolyl-valyl-O-methylaspartyl-[-2,6-difluorophenoxy]-methyl ketone, Boc-D-fmk and Z-VAD-fmk. “Sentence 17. The method of sentence 14, further comprising the step of supplying to the culture medium an Additive capable of increasing glutathione levels in the cell. “Sentence 18. The method of sentence 17, wherein the Additive capable of increasing glutathione levels in the cell is selected from one or more of Benzyl isothiocyanate, .beta.-naphthoflavone, coumarin, .alpha.-angelicalactone, disulfiram, indole-3-carbinol and indole-3-acetonitrile sulforaphane and dimethyl fumarate. “In a second embodiment, the invention may be described in part by the following numbered sentences: Sentence 19. A bioreactor comprising a source of Nitric Oxide gas or a source of a chemical capable of forming Nitric Oxide, wherein the bioreactor is configured for and adapted to provide the Nitric Oxide or chemical capable of forming Nitric Oxide to a cell culture medium within the bioreactor. “Sentence 20. The bioreactor of sentence 19, wherein the bioreactor is configured for and adapted to commercial scale production of a heterologous protein biomolecule of at least 1000 mg/L per a cell culture batch or per a single, substantially contiguous cell culture production run. “Sentence 21. The bioreactor of sentence 19 and/or 20, wherein the bioreactor further comprises: a) a Nitric Oxide sensor configured to and adapted for measuring the Nitric Oxide levels in the cell culture medium in the bioreactor, b) a Nitric Oxide gas injector and/or a chemical injector operably connected to a supply of a chemical source of Nitric Oxide, and c) a computer with a communication link to the Nitric Oxide sensor and electronically connected to the injector(s) of b), the computer specifically programmed to receive the Nitric Oxide sensor reading and modulate the injector(s) to maintain the Nitric Oxide concentration in the cell culture medium within a pre-determined concentration range and/or set point. “Sentence 22. The bioreactor of sentence 21, wherein the pre-determined concentration range is 0.1-10 nanomolar of Nitric Oxide solubilized in the cell culture medium. “Sentence 23. A method of operating the bioreactor of sentence 19, 20, and/or 21, comprising the step of providing the Nitric Oxide gas, and/or a chemical capable of forming Nitric Oxide, to the cell culture medium within the bioreactor of sentence 19, 20, and/or 21. “In a third embodiment, the invention may be described in part by the following numbered sentences: Sentence 24. A cell culture medium comprising a) a eukaryotic cell line that is capable of secreting a heterologous protein biomolecule, and b) an Additive capable of causing an increase in the amount of heterologous protein biomolecule secreted by the eukaryotic cell. “Sentence 25. The cell culture medium of sentence 24, wherein the Additive is selected from one or more of Nitric Oxide gas mixed with the cell culture medium, Nitric Oxide dissolved in the cell culture medium and Carbon Monoxide. “Sentence 26. The cell culture medium of sentence 24 and/or 25, wherein (a) the Additive is Nitric Oxide, (b) the eukaryotic cell line is a CHO cell line, and © the heterologous protein biomolecule is a monoclonal antibody. “Sentence 27. The method of sentence 24, 25, and/or 26, wherein the Nitric Oxide soluble in the cell culture medium is within a concentration range of 0.1-10 nanomolar of Nitric Oxide. “In a fourth embodiment, the invention may be described in part by the following numbered sentences: Sentence 28. A method of screening for a cell line that secretes a heterologous protein biomolecule and which is responsive to an Additive to result in an increased amount of secreted protein biomolecule, the method comprising the steps of: a) establishing a cell culture of the cell line that secretes the heterologous protein biomolecule, b) adding the Additive to the cell culture, c) measuring an amount of the secreted heterologous protein biomolecule protein biomolecule, and d) comparing the amount measured in step c) with an amount of secreted heterologous protein biomolecule representing a control condition lacking the addition of the Additive. “Sentence 29. The method of sentence 28, wherein the Additive is selected from one or more of Nitric Oxide gas, a chemical capable of forming Nitric Oxide, and Carbon Monoxide. “Sentence 30. The method of sentence 28 and/or 29, wherein (a) the Additive is selected from one or more of Nitric Oxide gas and a chemical capable of forming Nitric Oxide, (b) the cell line is a CHO cell line, and © the heterologous protein biomolecule is a monoclonal antibody.” The claims supplied by the inventors are: “1. A method of increasing protein biomolecule production comprising the steps of: a) culturing a cell that produces a protein biomolecule, b) supplying an Additive and/or a source of the Additive to the culture medium in an amount sufficient to (i) increase a total yield of the protein biomolecule secreted into the cell culture media and/or (ii) increase a specific cellular productivity of the protein biomolecule secreted into the cell culture media, c) wherein the increase of step b) is measured relative to: culturing under a prior set of conditions established for a regulatory approval for producing the protein biomolecule and/or culturing under substantially the same conditions except for supplying the Additive and/or a source of the Additive, wherein (a) the Additive is Nitric Oxide and/or a chemical source of Nitric Oxide, (b) the cell is a eukaryotic cell, and © the protein biomolecule is a heterologous protein biomolecule. “2. (canceled) “3. The method of claim 1, wherein the eukaryotic cell is a Chinese hamster ovary (CHO) cell and the heterologous protein biomolecule is a monoclonal antibody. “4. The method of claim 1, wherein the Nitric Oxide is gaseous and is substituted for a portion of a volume of another gas being fed into the bioreactor. “5. The method of claim 4, wherein the other gas being fed into the bioreactor includes Oxygen gas, Nitrogen gas and/or or Carbon Dioxide gas. “6. The method of claim 1, wherein the Nitric Oxide and/or the chemical source of Nitric Oxide are added in an amount sufficient to increase a total yield of the monoclonal antibody secreted into the cell culture media. “7. The method of claim 1, wherein the Nitric Oxide and/or the chemical source of Nitric Oxide are added in an amount sufficient to increase a specific cellular productivity of the monoclonal antibody secreted into the cell culture media. “8. The method of claim 1, wherein the Nitric Oxide soluble in the cell culture medium is at a concentration of 0.1-10 nanomolar. “9. The method of claim 8, wherein the Nitric Oxide is supplied to the cell culture medium as a sparged gas at a concentration of 200-400 ppm Nitric Oxide in the sparged gas. “10. The method of claim 1, wherein the eukaryotic cell is a CHO cell and the heterologous protein biomolecule is a monoclonal antibody. “11.-23. (canceled) “24. A cell culture medium comprising: a) a eukaryotic cell line that is capable of secreting a heterologous protein biomolecule, and b) an Additive capable of causing an increase in the amount of heterologous protein biomolecule secreted by the eukaryotic cell, wherein the Additive is selected from one or both of Nitric Oxide gas mixed with the cell culture medium and Nitric Oxide dissolved in the cell culture medium. “25. (canceled) “26. The cell culture medium of claim 24, wherein (a) the Additive is Nitric Oxide, (b) the eukaryotic cell line is a CHO cell line, and © the heterologous protein biomolecule is a monoclonal antibody. “27. The method of claim 26, wherein the Nitric Oxide soluble in the cell culture medium is within a concentration range of 0.1-10 nanomolar of Nitric Oxide. “28. A method of screening for a cell line that secretes a heterologous protein biomolecule and which is responsive to an Additive to result in an increased amount of secreted protein biomolecule, the method comprising the steps of: a) establishing a cell culture of the cell line that secretes the heterologous protein biomolecule, b) adding the Additive to the cell culture, c) measuring an amount of the secreted heterologous protein biomolecule protein biomolecule, and d) comparing the amount measured in step c) with an amount of secreted heterologous protein biomolecule representing a control condition lacking the addition of the Additive, wherein the Additive is selected from one or both of Nitric Oxide gas and a chemical capable of forming Nitric Oxide. “29. (canceled) “30. The method of claim 28, wherein (a) the Additive is selected from one or more of Nitric Oxide gas and a chemical capable of forming Nitric Oxide, (b) the cell line is a CHO cell line, and © the heterologous protein biomolecule is a monoclonal antibody.” (Our reports deliver fact-based news of research and discoveries from around the world.)



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