Aqueous solution containing glutathione salt

Abstract

The present disclosure provides pharmaceutical preparations, products, and methods related to ARINA-1 for use in treating patients with chronic inflammatory pulmonary diseases, such as lung transplants, cystic fibrosis (CF), non-CF bronchiectasis, chronic obstructive pulmonary disease (COPD), and other inflammatory pulmonary diseases.

Description

[Technical field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This application claims the benefit of U.S. Provisional Patent Application No. 63 / 296,405, filed January 4, 2022, which is incorporated by reference in its entirety.

[0002] FIELD OF THEINVENTION The present disclosure relates to the general field of chemistry and pharmaceutical preparations.In some aspects, the present disclosure relates to pharmaceutical preparations, such as those containing glutathione, suitable for pulmonary delivery, and the method of using said pharmaceutical preparations.In some aspects, the present disclosure provides pharmaceutical preparations and pharmaceutical products related to ARINA-1 for use in treating patients with chronic inflammatory lung diseases characterized by excess mucus and inflammation, such as lung transplants, cystic fibrosis (CF), non-CF bronchiectasis, chronic obstructive pulmonary disease (COPD), and other inflammatory lung diseases. [Background technology]

[0003] background Excessive mucus and inflammation are characteristic of chronic inflammatory lung diseases. Ion dysregulation contributes to excessive mucus production and chronic unregulated inflammation, both of which lead to lung injury and scarring that leads to lung failure. Ramsey et al., American Journal of Respiratory and Critical Care Medicine 201(6):661-670 (2020)(Non-Patent Document 1); Borok et al., Lancet 338:215-216 (1991)(Non-Patent Document 2); Behr et al., Transplantation 69:1856-1860 (2000)(Non-Patent Document 3). Glutathione is an important antioxidant ion that contributes to the structural integrity of mucus (Meldrum et al., Scientific Reports 8(1):Article number 5802 (2018) https: / / doi.org / 10.1038 / s41598-018-24223-3) and regulates inflammatory responses. Elferink, Immunobiology 184(1):25-36 (1991). Chronic inflammation also leads to increased airway acidity that contributes to lung injury. Glutathione dysregulation and increased airway acidity directly contribute to excessively viscous mucus with delayed or absent mucus ciliary clearance and resulting obstruction of small and medium airways. This creates an environment that is prone to bacterial colonization, chronic infection, and a feedback loop of inflammation.

[0004] Current treatments for chronic inflammatory lung disease include dornase alfa and hypertonic saline. Dornase alfa (Pulmozyme; Genentech Inc.) is an inhaled recombinant human deoxyribonuclease that cleaves extracellular DNA accumulated in CF mucus, Yang et al., Cochrane Database Syst Rev. 4:CD001127 (2016). Hypertonic saline is thought to hydrate the mucus layer through osmotic fluid transport. Donaldson et al., N Engl J Med. 354:241-250 (2006). However, improvements in lung function, exacerbations, and quality of life observed with these agents are modest and variable. Yang et al., Cochrane Database Syst Rev. 4:CD001127 (2016) (Non-patent document 8); Henke et al., Paediatr Respir Rev. 8:24-29 (2007) (Non-patent document 9).

[0005] ARINA-1 is a compounded pharmaceutical preparation used to treat CF and other chronic inflammatory lung diseases. ARINA-1 is prepared by dissolving bicarbonate, L-glutathione, and ascorbic acid in water. The aqueous solution thus obtained is nebulized before being administered to the lungs of a patient. See, for example, Adewale et al., Am J Respir Cell Mol Biol. 63:362-373 (2020); U.S. Patent Nos. 9,308,234 and 11,058,743; and U.S. Patent Application Nos. 2019 / 0351005 and 2020 / 0397849. There is a need in the art for stable pharmaceutical preparations and pharmaceutical products for use in treating pulmonary diseases. [Prior art documents] [Patent documents]

[0006]

Patent Document 1

Patent document 2

Patent Document 3

Patent document 4

Non-licensed literature

[0007] [Non-licensed document 1] Ramsey et al., American Journal of Respiratory and Critical Care Medicine 201(6):661-670 (2020)

Non-licensed Document 2

Non-licensed Document 4

Non-licensed Document 5

Non-licensed Document 6

Non-licensed Document 7

Non-licensed Document 8

[0008] overview The present disclosure provides stable pharmaceutical preparations and pharmaceutical products related to ARINA-1 for use in treating patients with chronic inflammatory lung diseases characterized by excess mucus and inflammation, such as lung transplants, cystic fibrosis (CF), non-CF bronchiectasis, chronic obstructive pulmonary disease (COPD), and other inflammatory lung diseases.

[0009] The formulation of ARINA-1 and related pharmaceutical preparations involves dissolving a bicarbonate salt, e.g., sodium bicarbonate, L-glutathione, and ascorbic acid in water. Applicants have discovered that such aqueous compositions are surprisingly stable when prepared, packaged, and stored under an atmosphere of carbon dioxide.

[0010] In one aspect, the present disclosure provides a preparation comprising an aqueous solution in a closed container having a headspace, (i) The aqueous solution is a solution of Formula I: Contains salt with TIFF2025502856000001.tif27128; (ii) the headspace atmosphere contains 90% or more by volume of carbon dioxide; and (iii) M + Na + , Li + , K + , or Cs + It is.

[0011] In another aspect, the disclosure provides a method of making a preparation comprising an aqueous solution comprising a salt having formula I, the method comprising: (i) To obtain an aqueous solution, L-glutathione, ascorbic acid, and M were mixed under a carbon dioxide atmosphere. + Na + , Li + , K + , or Cs + M + HCO3 - and dissolving the above in water for injection; (ii) transferring a portion of the aqueous solution to a container; (iii) blanketing the aqueous solution with carbon dioxide; and (iv) sealing the container with a stopper Includes.

[0012] In another aspect, the present disclosure provides a method for the preparation of a medicament for the treatment of cancer, comprising the steps of: mixing L-glutathione, ascorbic acid, and M under an atmosphere of carbon dioxide; + Na + , Li + , K + , or Cs + M + HCO3 - and in water for injection, an aqueous solution comprising the salt having formula I is provided. [Brief description of the drawings]

[0013] [Figure 1] 1 is a line graph showing pH versus time for an aqueous solution containing sodium bicarbonate, L-glutathione, and ascorbic acid prepared under a nitrogen atmosphere. [Diagram 2] 1 is a line graph showing pH versus time for an aqueous solution containing sodium bicarbonate, L-glutathione, and ascorbic acid formulated under an atmosphere of carbon dioxide. [Diagram 3] This is a partial NMR spectrum of an aqueous solution containing sodium bicarbonate, L-glutathione, and ascorbic acid prepared under an atmosphere of carbon dioxide. [Figure 4] FIG. 1 is a flow chart showing the formulation steps used to prepare ARINA-1 under an atmosphere of carbon dioxide. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0014] Detailed Description I. Preparations of the Present Disclosure In one aspect, referred to as "Aspect I," the present disclosure provides a preparation comprising an aqueous solution in a closed container having a headspace, (i) The aqueous solution is a solution of Formula I: Contains salt with TIFF2025502856000002.tif27128; (ii) the headspace atmosphere contains 90% or more by volume of carbon dioxide; and (iii) M + Na + , Li + , K + , or Cs + It is.

[0015] In another aspect, the aqueous solution of embodiment I has a pH of 6.0±0.4 at about 5° C. for 24 hours or more.

[0016] In another aspect, the aqueous solution of embodiment I has a pH of 6.0±0.3 at about 5° C. for 24 hours or more.

[0017] In another aspect, the aqueous solution of embodiment I has a pH of 6.0±0.2 at about 5° C. for 24 hours or more.

[0018] In another aspect, the aqueous solution of embodiment I has a pH of 6.0±0.1 at about 5° C. for 24 hours or more.

[0019] In another aspect, the aqueous solution of embodiment I comprises a compound of formula II: Further includes a salt having TIFF2025502856000003.tif23128.

[0020] In another aspect, the aqueous solution of embodiment I comprises a compound of formula III: Further included is a salt having TIFF2025502856000004.tif20128.

[0021] In another embodiment, the headspace atmosphere of embodiment I comprises 80% or more by volume carbon dioxide.

[0022] In another embodiment, the headspace atmosphere of embodiment I comprises 85% or more by volume carbon dioxide.

[0023] In another embodiment, the headspace atmosphere of embodiment I comprises 90% or more by volume carbon dioxide.

[0024] In another embodiment, the headspace atmosphere of embodiment I comprises 95% or more by volume carbon dioxide.

[0025] In another aspect, the aqueous solution of embodiment I contains from about 10% to about 20% by weight of the salt having formula I.

[0026] In another aspect, the aqueous solution of embodiment I contains from about 13% to about 17% by weight of the salt having formula I.

[0027] In another aspect, the aqueous solution of embodiment I comprises about 14.7 wt % of the salt having formula I.

[0028] In another aspect, the aqueous solution of embodiment I contains from about 5 wt % to about 15 wt % of the salt having formula II.

[0029] In another aspect, the aqueous solution of embodiment I contains from about 7 wt % to about 11 wt % of the salt having formula II.

[0030] In another aspect, the aqueous solution of embodiment I comprises about 9.1 wt% of the salt having formula II.

[0031] In another aspect, the aqueous solution of embodiment I has a density of about 1.13 g / L.

[0032] In another aspect, the aqueous solution of embodiment I is frozen.

[0033] In another aspect, M + In embodiment I, Na + It is.

[0034] In another aspect, M + In aspect I, Li + It is.

[0035] In another aspect, M + In aspect I, + It is.

[0036] In another aspect, M + In aspect I, Cs + It is.

[0037] In another embodiment, the preparation of embodiment I is packaged as a single unit dose. In another aspect, the single unit dose is in a sealed vial.

[0038] In another aspect, the preparation of embodiment I is sold, distributed, or administered as part of a pharmaceutical product.

[0039] In some aspects, the preparation of embodiment I can further comprise any one or more of the additional aspects disclosed herein.

[0040] II. Methods for Making the Preparation In another aspect, referred to as "Aspect II", the present disclosure provides a method of making the preparation of Aspect I (or Aspect I including one or more of the additional aspects disclosed above), the method comprising: (i) To obtain an aqueous solution, L-glutathione, ascorbic acid, and M were dissolved in water under carbon dioxide. + Na + , Li + , K + , or Cs + M + HCO3 - and dissolving the above in water for injection; (ii) transferring a portion of the aqueous solution to a container; (iii) blanketing the aqueous solution with carbon dioxide; and (iv) sealing the container with a stopper Includes.

[0041] In another embodiment, to obtain the aqueous solution of embodiment II, about 8 wt % to about 18 wt % L-glutathione, about 3 wt % to about 13 wt % ascorbic acid, and about 3 wt % to about 13 wt % M + HCO3 - is dissolved in about 62 wt% to about 82 wt% water for injection.

[0042] In another aspect, the aqueous solution of embodiment II has a pH of 6.0±0.4 at about 5° C. for 24 hours or more.

[0043] In another aspect, the aqueous solution of embodiment II has a pH of 6.0±0.3 at about 5° C. for 24 hours or more.

[0044] In another aspect, the aqueous solution of embodiment II has a pH of 6.0±0.2 at about 5° C. for 24 hours or more.

[0045] In another aspect, the aqueous solution of embodiment II has a pH of 6.0±0.1 at about 5° C. for 24 hours or more.

[0046] In another aspect, in embodiment II, M + Na + That is, M + HCO3 - is sodium bicarbonate.

[0047] In another aspect, in embodiment II, M + Li + That is, M + HCO3 - is lithium bicarbonate.

[0048] In another aspect, in embodiment II, M + is K + That is, M + HCO3 - is potassium bicarbonate.

[0049] In another aspect, in embodiment II, M + is Cs +That is, M + HCO3 - is cesium bicarbonate.

[0050] In another aspect, M + Na + To obtain the aqueous solution of embodiment II, about 11 wt % to about 15 wt % L-glutathione, about 5 wt % to about 9 wt % ascorbic acid, and about 5 wt % to about 9 wt % sodium bicarbonate are dissolved in about 68 wt % to about 76 wt % water.

[0051] In another aspect, M + Na + To obtain an aqueous solution of embodiment II, about 13.0 wt. % L-glutathione, about 7.6 wt. % ascorbic acid, and about 7.3 wt. % sodium bicarbonate are dissolved in about 72.0 wt. % water.

[0052] In some aspects, the method of embodiment II can further include any one or more of the additional aspects disclosed herein.

[0053] III. Product by Process In another aspect, referred to as "Aspect III," the present disclosure provides a method for preparing a medicament for use in a pharmaceutical composition comprising: + Na + , Li + , K + , or Cs + M + HCO3 - and in water for injection, An aqueous solution containing a salt having TIFF2025502856000005.tif27128 is provided.

[0054] In another aspect, to obtain the aqueous solution of embodiment III, about 8 wt % to about 18 wt % L-glutathione, about 3 wt % to about 13 wt % ascorbic acid, and about 3 wt % to about 13 wt % M + HCO3 - is dissolved in about 62 wt% to about 82 wt% water for injection.

[0055] In another aspect, the aqueous solution of embodiment III has a pH of 6.0±0.4 at about 5° C. for 24 hours or more.

[0056] In another aspect, the aqueous solution of embodiment III has a pH of 6.0±0.3 at about 5° C. for 24 hours or more.

[0057] In another aspect, the aqueous solution of embodiment III has a pH of 6.0±0.2 at about 5° C. for 24 hours or more.

[0058] In another aspect, the aqueous solution of embodiment III has a pH of 6.0±0.1 at about 5° C. for 24 hours or more.

[0059] In another aspect, the aqueous solution of embodiment III comprises a compound of formula II: Further includes a salt having TIFF2025502856000006.tif23128.

[0060] In another aspect, the aqueous solution of embodiment III comprises a compound of formula III: Further included is a salt having TIFF2025502856000007.tif20128.

[0061] In another aspect, in embodiment III, M + Na + That is, M + HCO3 - is sodium bicarbonate.

[0062] In another aspect, in embodiment III, M + Li + That is, M + HCO3 - is lithium bicarbonate.

[0063] In another aspect, in embodiment III, M + is K + That is, M + HCO3 - is potassium bicarbonate.

[0064] In another aspect, in embodiment III, M + is Cs + That is, M + HCO3 - is cesium bicarbonate.

[0065] In another aspect, to obtain the aqueous solution of embodiment III, about 11 wt % to about 15 wt % L-glutathione, about 5 wt % to about 9 wt % ascorbic acid, and about 5 wt % to about 9 wt % sodium bicarbonate are dissolved in about 68 wt % to about 76 wt % water.

[0066] In another aspect, about 13.0 wt % L-glutathione, about 7.6 wt % ascorbic acid, and about 7.3 wt % sodium bicarbonate are dissolved in about 72.0 wt % water to obtain the aqueous solution of embodiment III.

[0067] In some aspects, the aqueous solution of embodiment III can further comprise any one or more of the additional aspects disclosed herein.

[0068] IV. Method of Use The preparations or aqueous solutions disclosed herein are useful for treating, alleviating symptoms of, or preventing various diseases, conditions, or disorders, including those described in Sections I-II (e.g., embodiments I and III and further aspects thereof). In particular, these preparations and aqueous solutions are useful in therapeutic methods for treating, alleviating symptoms of, or preventing diseases, conditions, or disorders in which administration of L-glutathione or a salt thereof and / or ascorbic acid or a salt thereof is beneficial.

[0069] In one aspect, a method of treating or preventing a lung or airway disorder in a subject in need thereof comprises administering to the subject a therapeutically effective amount of a preparation described in Section I or an aqueous solution described in Section III, e.g., via inhalation.

[0070] In one aspect, the lung or airway disorder is a chronic inflammatory lung disease, pulmonary fibrosis, pulmonary vasculitis, pulmonary sarcoidosis, inflammation and / or infection associated with lung transplantation, acute or chronic lung rejection, chronic lung allograft dysfunction (CLAD, pulmonary arterial hypertension, bronchitis, sinusitis, asthma, cystic fibrosis, bacterial infection, fungal infection, parasitic infection, viral infection, chronic obstructive pulmonary disease (COPD), bronchiolitis obliterans syndrome (BOS), primary ciliary dyskinesia, and / or primary pulmonary fibrosis. (PCD), pulmonary alveolar proteinosis, idiopathic pulmonary fibrosis, eosinophilic pneumonia, eosinophilic bronchitis, acute respiratory distress syndrome (ARDS), inflammation and / or infection associated with mechanical ventilation, ventilator-associated pneumonia, complications associated with short-term or long-term placement of a tracheostomy opening, asbestos-related airway disorders or diseases, dust-related airway disorders or diseases, silicosis, and radiation- or chemical-related airway diseases or disorders, and any combination thereof.

[0071] In another aspect, the lung or airway disorder is selected from the group consisting of chronic inflammatory lung disease, inflammation and / or infection associated with lung transplantation, acute or chronic lung rejection, chronic pulmonary allograft dysfunction (CLAD), asthma, cystic fibrosis, and chronic obstructive pulmonary disease (COPD), complications associated with short-term or long-term placement of a tracheostomy opening, and any combination thereof. In another embodiment, the method further comprises administering an additional therapeutic agent to the subject.

[0072] In another aspect, lung or airway damage occurs collaterally from radiation and / or chemotherapy in subjects with cancer.

[0073] In another aspect, the disclosure provides a method of treating or preventing an infection in the respiratory tract of a subject in need thereof, comprising administering to the subject a therapeutically effective amount of a preparation described in Section I. In another embodiment, a preparation described in Section I is administered to the subject in combination with one or more antibiotics. The antibiotics may be administered locally to the lungs and / or systemically.

[0074] In another aspect, the disclosure provides a method of treating or preventing inflammation in the airways of a subject in need thereof, comprising administering to the subject a preparation described in Section I.

[0075] In another aspect, the disclosure provides a method of treating or preventing a disease or disorder in a mucosal tissue in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of a preparation described in Section I. Non-limiting examples of mucosal tissues include the mouth, nose, eyes, ears, upper respiratory tract, lower respiratory tract, gastrointestinal tract, vagina, rectum, and urethra.

[0076] In another aspect, the disclosure provides a method of treating or preventing a disease or disorder associated with a mucosa in a subject in need thereof, comprising administering to the subject a therapeutic amount of a preparation described in Section I to a suitable mucosa. In one embodiment, the mucosa is the lung, such as the intermediate bronchioles or the deep lung (alveolar region), and in another embodiment, the mucosa is one or more of the eye, mouth, nose, rectum, urogenital tract, and vagina.

[0077] In another aspect, the preparations described in Section I are used to treat or prevent bronchiolitis obliterans and military-related lung injury, i.e., lung injury in military personnel with airway damage secondary to unknown exposure.

[0078] Therapeutic methods of the disclosure include administering to a subject in need thereof, e.g., a human patient, a therapeutically effective amount of a preparation described in Section I. Whether such treatment is indicated will vary on an individual case basis and is subject to a medical evaluation (diagnosis) taking into account the signs, symptoms and / or dysfunction present, the risk of developing the particular signs, symptoms and / or dysfunction, and other factors.

[0079] In some aspects, the preparations described in Section I are nebulized prior to administration to a subject.

[0080] V. Definition The term "ARINA-1" refers to a pharmaceutical preparation obtained by formulating L-glutathione, ascorbic acid, and sodium bicarbonate in water to form an aqueous solution. The chemicals present in ARINA-1 when prepared and maintained under an atmosphere of CO2 can include sodium L-ascorbate, sodium L-glutathione / bicarbonate, and sodium L-glutathione, as shown in Scheme 1. Scheme 1 TIFF2025502856000008.tif98159

[0081] The stoichiometry of the components of ARINA-1 is shown in Table 1.

[0082] [Table 1]

[0083] The term "pharmaceutical product" refers to a product suitable for human use. In some aspects, a pharmaceutical product is suitable for human use and is subject to regulation by the U.S. Food and Drug Administration (FDA) or a foreign equivalent to the FDA, including, but not limited to, the European Medicines Agency (EMA), the National Medical Products Administration of China (NMPA), and the Ministry of Health, Labor and Welfare of Japan (MHLW).

[0084] The term "preparation" refers to one or more drugs intended for human use, either (i) in their finished dosage form or (ii) used in the preparation, formulation, and / or administration of the finished dosage form.

[0085] The term "container" refers to a pharma- ceutically acceptable container that includes a chamber suitable for containing an aqueous solution that includes one or more drug products. The "closed end" of the container refers to the end of the chamber that does not have an opening. The term "open end" of the container refers to the end of the chamber opposite the closed end. Exemplary containers include, but are not limited to, vials, syringes, cartridges, bags, such as polypropylene and polyurethane bags, and ampoules. In one embodiment, the container is a vial.

[0086] As used herein, the term "plug" refers to any article capable of preventing aqueous solutions and gases, such as carbon dioxide, from escaping through the open end of a container.

[0087] As used herein, a "sealed container" refers to a container fitted with a stopper.

[0088] As used herein, the term "headspace" refers to the area within the chamber of a container that is between the aqueous solution and the stopper when the open end of the container is oriented away from the pull of gravity.

[0089] The term "atmosphere" as used herein refers to one or more layers of gas in the head space of a closed container. These gases include, but are not limited to, oxygen, nitrogen, argon, and carbon dioxide. In one embodiment, the atmosphere in the pharmaceutical product of the present disclosure comprises 75% or more carbon dioxide by volume, for example, 80% or more carbon dioxide, 85% or more carbon dioxide, 90% or more carbon dioxide, 95% or more carbon dioxide, or 99% or more carbon dioxide.

[0090] The term "wt %" as used herein in reference to aqueous solutions refers to the weight of one component divided by the combined weight of all components (including, for example, water for injection) multiplied by 100. For example, 161 g of L-glutathione sodium / bicarbonate, i.e., M + Na + and 99 g of sodium ascorbate, i.e., M + Na +The wt% of L-glutathione sodium / bicarbonate in an aqueous solution containing the compound of formula II, wherein:

[0091] The terms "a" and "an" refer to one or more than one.

[0092] As used herein, the term "about" includes the stated number plus or minus 10%, so "about 10" means 9 to 11. EXAMPLES

[0093] General method material L-Ascorbic acid (Lot #1200851004) and reduced L-glutathione (Lot #B200465) used in the 2 L scale experiments were procured from CSPC Weisheng Pharmaceutical (Shijiazhuang) Co. and Shandong Jincheng Biopharmaceutical Co., respectively. L-Ascorbic acid (Product #A92902) and reduced L-glutathione (Product #G4251) used in the 0.75 L scale experimental runs with carbon dioxide blanketing were purchased from Millipore Sigma Co. Sodium bicarbonate (Product #S6014) for all tests was purchased from Millipore Sigma Co. Deionized water was supplied from Millipore's water system. All other raw materials and solvents used to analyze the products were purchased from commercial sources and used as received.

[0094] NMR equipment NMR was used to determine the titration concentrations of L-ascorbic acid, reduced L-glutathione, L-glutathione disulfide and any unknown impurities in the demonstration run. 1 H NMR spectra were acquired at 25 °C using a Bruker Avance III 500 MHz NMR system equipped with a 5 mm triple resonance cryoprobe. Deuterium oxide was used as the NMR solvent, and the resonances from calcium formate were used as the reference to establish chemical shifts and integrals. NMR spectral data were processed using MNova software.

[0095] Example 1 Preparation of ARINA-1 under N2 A 2 L scale preparation of ARINA-1 referenced in Table 2 was carried out in a jacketed 4-neck glass reactor equipped with an overhead stirrer, thermometer, gas inlet adapter, port for solids loading, and a second adapter to limit the escape of nitrogen from the flask via a Claisen adapter. Nitrogen was charged to the flask via a flowmeter capable of measuring a flow rate of 0.05-0.5 L / min. Temperature control of the reaction was achieved by recirculating a water / glycol mixture from the vessel jacket to a Haake EZ-Cool 80 circulating heater / cooler.

[0096] [Table 2]

[0097] At the end of L-glutathione neutralization, the pH was monitored and the increase in pH was measured to be approximately +0.1 pH units per hour at the temperatures (° C.) shown in Table 3 and FIG.

[0098] [Table 3]

[0099] Example 2 Preparation of ARINA-1 under CO2 The preparation of the 750 mL scale ARINA-1 referenced in Table 4 was carried out in a 1 L jacketed 4-neck glass reactor equipped with an overhead stirrer, thermometer, gas inlet adapter, and a port for solids loading. Carbon dioxide was charged to the reactor via a gas flow meter measuring 0.05-0.5 L / min. Temperature control of the reaction was achieved by recirculating a water / glycol mixture between the vessel jacket and a Haake EZ-Cool 80 circulating heater / cooler. The steps of the formulation process are outlined in Figure 4.

[0100] [Table 4]

[0101] Process description: 1. 600 g of deionized water was added to the reactor. This amount of water plus the rinse introduced in step 6 gave a product density of 1.12-1.13 g / mL. Agitation was commenced with the overhead stirrer speed set to approximately 90-100 rpm. The water temperature for the demonstration experiment was 21°C.

[0102] 2. 63.0 g of sodium bicarbonate was added and stirring was continued for approximately 10 minutes until most of the sodium bicarbonate was dissolved.

[0103] 3. The air in the headspace of the flask was completely replaced with gaseous carbon dioxide from a cylinder source equipped with an appropriate gas flow regulator.

[0104] 4. Charge 66.0 g of L-ascorbic acid to the flask at a rate that ensures controlled evolution of carbon dioxide gas. Once the solids have been added, the reaction is complete within 2-3 minutes to give a clear solution, after which step 5 can begin.

[0105] 5. 112.5 g of reduced L-glutathione was charged to the flask at a rate that ensured controlled evolution of carbon dioxide gas. The reaction was complete within 4-5 minutes after the last solid was added, yielding a clear solution.

[0106] 6. Weigh out 21.75 g of deionized water and use it to wash down any remaining solids from the solids addition funnel and / or the walls of the top of the vessel where any solids may be present. The agitation speed was increased significantly and held for 10-15 seconds to dislodge any solids or liquids adhering to the vessel walls to obtain a completely homogenous mixture.

[0107] 7. A slower stirring speed was resumed (but still maintained sufficient stirring speed to promote heat transfer) and the solution was immediately cooled to 2-6°C.

[0108] Upon completion of L-glutathione neutralization, the pH was monitored at the temperatures (° C.) shown in Table 5 and FIG.

[0109] [Table 5]

[0110] The problem with continuous nitrogen sparging or sweeping in Example 1 is that residual CO2 is swept out of the system by the nitrogen flow, which in turn promotes the decomposition of carbonate formed by the neutralization reaction. This causes the pH to rise above the target range of pH 5.8-6.4. Experiments using a slow nitrogen sweep showed that the pH of the formulated product rose from 6.0 to 7.3 after approximately 18 hours of post-reaction inactivation (data not shown).

[0111] Henry's gas law states that the amount of gas dissolved in a liquid is proportional to its partial pressure above the liquid. Under conditions of slow, continuous nitrogen sweep, the partial pressure of CO2 in the headspace above the product solution is maintained near zero. However, when gaseous CO2 is used to inert the headspace, the pressure of CO2 will be 1 atmosphere, which should provide the driving force to reverse the loss of CO2 and establish an equilibrium that stabilizes the pH. This principle was demonstrated in this example.

[0112] The reduced L-glutathione is neutralized by sodium bicarbonate and is an acid. Gaseous CO2 evolution would not be expected to cease until equilibrium is established between the CO2 dissolved in the solution and the CO2 in the headspace. As with the previous run under nitrogen, for example, the pH rose to about 5.95 to 6.10 after dissolution of the solids, but gas evolution was still evident while the solution was cooled to 2-6°C. Equilibrium was established when the target pH range of 6.0-6.1 was reached. Raising and lowering the temperature over the next 4 hours of the experiment had no significant effect on the pH. A pH of 6.0 was again obtained after maintaining the solution under a slight positive pressure of CO2 for 20+ hours. The solution was sampled for NMR analysis, yielding assays of 99.7% and 99.0% for L-ascorbic acid and reduced L-glutathione, respectively. See Figure 3. The disulfide concentration was 0.25% and there were no unexpected resonances indicating unknown degradants at or above 0.2% concentration. Thus, the use of a CO2 blanket unexpectedly stabilized the pH in its target range, as was necessary to keep the product pH (and CO2 content) constant throughout the vial fill cycle while excluding oxygen and controlling disulfide formation.

[0113] All inputs to this example were carefully measured by weight to allow for analysis of the actual product CO2 content corresponding to a pH of 6.1. As shown in the following stoichiometric calculation for a 1 L preparation of ARINA-1, if all the CO2 was expelled, the product would be expected to weigh 1107.6 g, but if all the CO2 was unexpectedly retained in solution, the product would weigh 1151.0 g. A 500 mL portion of this example preparation was weighed and found to be 1125.8 g / L. TIFF2025502856000014.tif135140

[0114] This corresponds to a carbonate content of 18.2 g / L (0.29 moles), or about 30% of the molar equivalent of the two APIs combined. Since the solubility of CO2 in water at 0°C and 1 atm of CO2 is 0.034 g / L, one can speculate that the solutes present serve to stabilize the carbonate in an unexpected manner. Reduced L-glutathione sodium salt is most likely responsible for this stabilization, as it can form carbonate complexes. TIFF2025502856000015.tif40163

[0115] Although the compounds, compositions, methods, formulations, and products have been fully described herein, those skilled in the art will recognize that the same can be practiced within a wide and equivalent range of conditions, formulations, and other parameters without affecting the scope of the compounds, compositions, methods, formulations, and products provided herein or any aspect thereof. All patents, patent applications, and publications cited herein are hereby incorporated by reference in their entirety.

Claims

1. 1. A preparation comprising an aqueous solution in a sealed container having a headspace, (i) the aqueous solution is a compound of Formula I: including salts having the formula: (ii) the headspace atmosphere contains 90% or more by volume of carbon dioxide; and (iii)M + Na + , Li + , K. + , or Cs + That is, The preparation.

2. 10. The preparation of claim 1, wherein the aqueous solution has a pH of 6.0±0.4 at about 5° C. for 24 hours or more.

3. 3. The preparation of claim 2, wherein the aqueous solution has a pH of 6.0±0.1 at about 5° C. for 24 hours or more.

4. The aqueous solution comprises a compound of Formula II:

10. The formulation of claim 1, further comprising a salt having the formula:

5. The aqueous solution comprises a compound of Formula III:

10. The formulation of claim 1, further comprising a salt having the formula:

6. 10. The preparation of claim 1, wherein the headspace atmosphere comprises 95% or more by volume of carbon dioxide.

7. 10. The formulation of claim 1, wherein the aqueous solution comprises about 10% to about 20% by weight of the salt having Formula I.

8. 8. The preparation of claim 7, wherein the aqueous solution comprises from about 13% to about 17% by weight of the salt having Formula I.

9. 9. The preparation of claim 8, wherein the aqueous solution comprises about 14.7 wt% of the salt having formula I.

10. 5. The formulation of claim 4, wherein the aqueous solution comprises from about 5 wt% to about 15 wt% of the salt having formula II.

11. 11. The formulation of claim 10, wherein the aqueous solution comprises from about 7 wt% to about 11 wt% of the salt having formula II.

12. 12. The preparation of claim 11, wherein the aqueous solution comprises about 9.1 wt% of the salt having formula II.

13. 10. The preparation of claim 1, wherein the aqueous solution has a density of about 1.13 g / L.

14. 10. The preparation of claim 1, wherein the aqueous solution is frozen.

15. M + Na + 2. The preparation of claim 1, wherein

16. 10. The preparation of claim 1, packaged as a single unit dose.

17. 10. The preparation of claim 1, which is a pharmaceutical product.

18. (i) dissolving L-glutathione, ascorbic acid and sodium bicarbonate in water for injection under carbon dioxide to obtain an aqueous solution; (ii) transferring a portion of the aqueous solution to a container; (iii) blanketing the aqueous solution with carbon dioxide; and (iv) sealing the container with a stopper Including, To obtain the aqueous solution, about 8 wt% to about 18 wt% L-glutathione, about 3 wt% to about 13 wt% ascorbic acid, and about 3 wt% to about 13 wt% sodium bicarbonate are dissolved in about 62 wt% to about 82 wt% water for injection; 16. A method for producing the preparation of claim 15.

19. 20. The method for producing the preparation of claim 18, wherein about 11 wt % to about 15 wt % L-glutathione, about 5 wt % to about 9 wt % ascorbic acid, and about 5 wt % to about 9 wt % sodium bicarbonate are dissolved in about 68 wt % to about 76 wt % water to obtain the aqueous solution.

20. 20. The method for producing the preparation of claim 19, wherein about 13.0 wt. % L-glutathione, about 7.6 wt. % ascorbic acid, and about 7.3 wt. % sodium bicarbonate are dissolved in about 72.0 wt. % water to obtain the aqueous solution.

21. Formula I: prepared by dissolving L-glutathione, ascorbic acid, and sodium bicarbonate in water for injection under an atmosphere of carbon dioxide. An aqueous solution comprising a salt having the formula: M + Na + and To obtain the aqueous solution, about 8 wt % to about 18 wt % L-glutathione, about 3 wt % to about 13 wt % ascorbic acid, and about 3 wt % to about 13 wt % sodium bicarbonate are dissolved in about 62 wt % to about 82 wt % water; The aqueous solution.

22. 22. The aqueous solution of claim 21, having a pH of 6.0±0.4 at about 5° C. for 24 hours or more.

23. 23. The aqueous solution of claim 22, having a pH of 6.0±0.1 at about 5° C. for 24 hours or more.

24. M + Na + Formula II:

22. The aqueous solution of claim 21, further comprising a salt having the formula:

25. M + Na + Formula III:

22. The aqueous solution of claim 21, further comprising a salt having the formula:

26. 22. The aqueous solution of claim 21, wherein about 11 wt% to about 15 wt% L-glutathione, about 5 wt% to about 9 wt% ascorbic acid, and about 5 wt% to about 9 wt% sodium bicarbonate are dissolved in about 68 wt% to about 76 wt% water to obtain the aqueous solution.

27. 27. The aqueous solution of claim 26, wherein about 13.0 wt% L-glutathione, about 7.6 wt% ascorbic acid, and about 7.3 wt% sodium bicarbonate are dissolved in about 72.0 wt% water to obtain the aqueous solution.

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