# Patent

## Title
Delamination resistant pharmaceutical glass containers containing active pharmaceutical ingredients

## Abstract

The present invention is based, at least in part, on the identification of a pharmaceutical container formed, at least in part, of a glass composition which exhibits a reduced propensity to delaminate, i.e., a reduced propensity to shed glass particulates. As a result, the presently claimed containers are particularly suited for storage of pharmaceutical compositions and, specifically, a pharmaceutical solution comprising a pharmaceutically active ingredient, for example, PEDIARIX® (Diphtheria and Tetanus Toxoids and Acellular Pertussis Adsorbed, Hepatitis B (Recombinant) and Inactivated Poliovirus Vaccine), HAVRIX® (Hepatitis A Vaccine), ENGERIX-B® (Hepatitis B Vaccine (Recombinant)), TWINRIX® (Hepatitis A & Hepatitis B (Recombinant) Vaccine), EPERZAN® (albiglutide), MAGE-A3 Antigen-Specific Cancer Immunotherapeutic (astuprotimut-R), GSK2402968 (drisapersen), and HZ/su (herpes zoster vaccine).

## Background
CROSS REFERENCE TO RELATED APPLICATIONS
The present application is related to U.S. Provisional Application No. 61/815,648, filed Apr. 24, 2013, entitled “Delamination Resistant Pharmaceutical Glass Containers Containing Active Pharmaceutical Ingredients”, the entirety of which is hereby incorporated by reference herein.
SEQUENCE LISTING
The instant application contains a Sequence Listing which has been submitted electronically in ASCII format and is hereby incorporated by reference in its entirety. Said ASCII copy, created on Apr. 22, 2014, is named 122467-02002_SL.txt and is 787 bytes in size.
FIELD OF THE INVENTION
The present specification generally relates to pharmaceutical containers and, more specifically, to chemically and mechanically durable pharmaceutical containers that are delamination resistant and formed, at least in part, of a glass composition.
BACKGROUND
The design of a packaged pharmaceutical composition generally seeks to provide an active pharmaceutical ingredient (API) in a suitable package that is convenient to use, that maintains the stability of the API over prolonged storage, and that ultimately allows for the delivery of efficacious, stable, active, nontoxic and nondegraded API.
Most packaged formulations are complex physico-chemical systems, through which the API is subject to deterioration by a variety of chemical, physical, and microbial reactions. Interactions between drugs, adjuvants, containers, and/or closures may occur, which can lead to the inactivation, decomposition and/or degradation of the API.
Historically, glass has been used as the preferred material for packaging pharmaceuticals because of its hermeticity, optical clarity and excellent chemical durability relative to other materials. Specifically, the glass used in pharmaceutical packaging must have adequate chemical durability so as not to affect the stability of the pharmaceutical compositions contained therein. Glasses having suitable chemical durability include those glass compositions within the ASTM standard ‘Type 1B’ glass compositions which have a proven history of chemical durability.
However, use of glass for such applications is limited by the mechanical performance of the glass. Specifically, in the pharmaceutical industry, glass breakage is a safety concern for the end user as the broken package and/or the contents of the package may injure the end user. Further, non-catastrophic breakage (i.e., when the glass cracks but does not break) may cause the contents to lose their sterility which, in turn, may result in costly product recalls.
One approach to improving the mechanical durability of the glass package is to thermally temper the glass package. Thermal tempering strengthens glass by inducing a surface compressive stress during rapid cooling after forming. This technique works well for glass articles with flat geometries (such as windows), glass articles with thicknesses >2 mm, and glass compositions with high thermal expansion. However, pharmaceutical glass packages typically have complex geometries (vial, tubular, ampoule, etc.), thin walls ( ̃1-1.5 mm), and are produced from low expansion glasses (30-55×10−7K−1) making glass pharmaceutical packages unsuitable for strengthening by thermal tempering.
Chemical tempering also strengthens glass by the introduction of surface compressive stress. The stress is introduced by submerging the article in a molten salt bath. As ions from the glass are replaced by larger ions from the molten salt, a compressive stress is induced in the surface of the glass. The advantage of chemical tempering is that it can be used on complex geometries, thin samples, and is relatively insensitive to the thermal expansion characteristics of the glass substrate. However, glass compositions which exhibit a moderate susceptibility to chemical tempering generally exhibit poor chemical durability and vice-versa.
Finally, glass compositions commonly used in pharmaceutical packages, e.g., Type 1a and Type 1b glass, further suffer from a tendency for the interior surfaces of the pharmaceutical package to shed glass particulates or “delaminate” following exposure to pharmaceutical solutions. Such delamination often destabilizes the active pharmaceutical ingredient (API) present in the solution, thereby rendering the API therapeutically ineffective or unsuitable for therapeutic use.
Delamination has caused the recall of multiple drug products over the last few years (see, for example, Reynolds et al., (2011) BioProcess International 9(11) pp. 52-57). In response to the growing delamination problem, the U.S. Food and Drug Administration (FDA) has issued an advisory indicating that the presence of glass particulate in injectable drugs can pose a risk.
The advisory states that, “[t]here is potential for drugs administered intravenously that contain these fragments to cause embolic, thrombotic and other vascular events; and subcutaneously to the development of foreign body granuloma, local injections site reactions and increased immunogenicity.”
Accordingly, a recognized need exists for alternative glass containers for packaging of pharmaceutical compositions which exhibit a reduced propensity to delaminate.
SUMMARY
In one aspect, the present invention is directed to a delamination resistant pharmaceutical container formed, at least in part, of a glass composition including from about 70 mol. % to about 80 mol. % SiO2; from about 3 mol. % to about 13 mol. % alkaline earth oxide; X mol. % Al2O3; and Y mol. % alkali oxide, wherein the alkali oxide includes Na2O in an amount greater than about 8 mol. %, wherein the ratio of Y:X is greater than 1, and the glass composition is free of boron and compounds of boron.
In one embodiment, the SiO2 is present in an amount less than or equal to 78 mol. %.
In one embodiment, the amount of the alkaline earth oxide is greater than or equal to about 4 mol. % and less than or equal to about 8 mol. %. In a particular embodiment, the alkaline earth oxide includes MgO and CaO and has a ratio (CaO (mol. %)/(CaO (mol. %)+MgO (mol. %))) that is less than or equal to 0.5. In a particular embodiment, the alkaline earth oxide includes from about 0.1 mol. % to less than or equal to about 1.0 mol. % CaO. In a particular embodiment, the alkaline earth oxide includes from about 3 mol. % to about 7 mol. % MgO.
In another embodiment, the alkali oxide includes greater than or equal to about 9 mol. % Na2O and less than or equal to about 15 mol. % Na2O. In another embodiment, the alkali oxide further includes K2O in an amount less than or equal to about 3 mol. %. In a particular embodiment, the alkali oxide includes K2O in an amount greater than or equal to about 0.01 mol. % and less than or equal to about 1.0 mol. %.
In one embodiment, X is greater than or equal to about 2 mol. % and less than or equal to about 10 mol. %. In a particular embodiment, the ratio of Y:X is less than or equal to 2. In a particular embodiment, the ratio of Y:X is greater than or equal to 1.3 and less than or equal to 2.0.
In another embodiment, the glass composition is free of phosphorous and compounds of phosphorous.
In one embodiment, the glass composition has a type HGB1 hydrolytic resistance according to ISO 719. Alternatively or in addition, the glass composition has a type HGA1 hydrolytic resistance according to ISO 720 after ion exchange strengthening. Alternatively or in addition, the glass composition has a type HGA1 hydrolytic resistance according to ISO 720 before and after ion exchange strengthening. Alternatively or in addition, the glass composition has at least a class S3 acid resistance according to DIN 12116. Alternatively or in addition, the glass composition has at least a class A2 base resistance according to ISO 695.
In one embodiment, the glass composition is ion exchange strengthened.
In another embodiment, the composition further includes a compressive stress layer with a depth of layer greater than or equal to 10 μm and a surface compressive stress greater than or equal to 250 MPa.
In another aspect, the present invention provides a delamination resistant pharmaceutical container formed, at least in part, of a glass composition including from about 72 mol. % to about 78 mol. % SiO2; from about 4 mol. % to about 8 mol. % alkaline earth oxide; X mol. % Al2O3, wherein X is greater than or equal to about 4 mol. % and less than or equal to about 8 mol. %; and Y mol. % alkali oxide, wherein the alkali oxide includes Na2O in an amount greater than or equal to about 9 mol. % and less than or equal to about 15 mol. %, wherein the ratio of Y:X is greater than 1, and the glass composition is free of boron and compounds of boron.
In a particular embodiment, the ratio of Y:X is less than or equal to about 2. In a particular embodiment, the ratio of Y:X is greater than or equal to about 1.3 and less than or equal to about 2.0.
In one embodiment, the alkaline earth oxide includes MgO and CaO and has a ratio (CaO (mol. %)/(CaO (mol. %)+MgO (mol. %))) less than or equal to 0.5.
In another embodiment, the alkali oxide includes K2O in an amount greater than or equal to about 0.01 mol. % and less than or equal to about 1.0 mol. %.
In another aspect, the present invention provides a delamination resistant pharmaceutical container formed, at least in part, of a glass composition including from about 68 mol. % to about 80 mol. % SiO2; from about 3 mol. % to about 13 mol. % alkaline earth oxide; X mol. % Al2O3; Y mol. % alkali oxide, wherein the alkali oxide includes Na2O in an amount greater than about 8 mol. %; and B2O3, wherein the ratio (B2O3 (mol. %)/(Y mol. %−X mol. %) is greater than 0 and less than 0.3, and the ratio of Y:X is greater than 1.
In one embodiment, the amount of SiO2 is greater than or equal to about 70 mol. %.
In one embodiment, the amount of alkaline earth oxide is greater than or equal to about 4 mol. % and less than or equal to about 8 mol. %. In a particular embodiment, the alkaline earth oxide includes MgO and CaO and has a ratio (CaO (mol. %)/(CaO (mol. %)+MgO (mol. %))) less than or equal to 0.5. In a particular embodiment, the alkaline earth oxide includes CaO in an amount greater than or equal to about 0.1 mol. % and less than or equal to about 1.0 mol. %. In a particular embodiment, the alkaline earth oxide includes from about 3 mol. % to about 7 mol. % MgO.
In one embodiment, the alkali oxide is greater than or equal to about 9 mol. % Na2O and less than or equal to about 15 mol. % Na2O. In a particular embodiment, the alkali oxide further includes K2O in a concentration less than or equal to about 3 mol. %. In another embodiment, the alkali oxide further includes K2O in a concentration greater than or equal to about 0.01 mol. % and less than or equal to about 1.0 mol. %.
In another embodiment, the pharmaceutical container has a ratio (B2O3 (mol. %)/(Y mol. %−X mol. %) less than 0.2. In a particular embodiment, the amount of B2O3 is less than or equal to about 4.0 mol. %. In another embodiment, the amount of B2O3 is greater than or equal to about 0.01 mol. %.
In one embodiment, X is greater than or equal to about 2 mol. % and less than or equal to about 10 mol. %. In a particular embodiment, the ratio of Y:X is less than or equal to 2. In another embodiment, the ratio of Y:X is greater than 1.3.
In one embodiment, the glass composition is free of phosphorous and compounds of phosphorous.
In one embodiment, the glass composition has a type HGB1 hydrolytic resistance according to ISO 719. Alternatively or in addition, the glass composition has a type HGA1 hydrolytic resistance according to ISO 720 after ion exchange strengthening. Alternatively or in addition, the glass composition has a type HGA1 hydrolytic resistance according to ISO 720 before and after ion exchange strengthening. Alternatively or in addition, the glass composition has at least a class S3 acid resistance according to DIN 12116. Alternatively or in addition, the glass composition has at least a class A2 base resistance according to ISO 695.
In one embodiment, the glass composition is ion exchange strengthened.
In another embodiment, the composition further includes a compressive stress layer with a depth of layer greater than or equal to 10 μm and a surface compressive stress greater than or equal to 250 MPa.
In one embodiment of any of the foregoing aspects of the invention, the pharmaceutical container further includes a pharmaceutical composition having an active pharmaceutical ingredient. In a particular embodiment, the pharmaceutical composition includes a citrate or phosphate buffer, for example, sodium citrate, SSC, monosodium phosphate or disodium phosphate. Alternatively or in addition, the pharmaceutical composition has a pH between about 7 and about 11, between about 7 and about 10, between about 7 and about 9, or between about 7 and about 8.
In one embodiment of any of the foregoing aspects of the invention, the active pharmaceutical ingredients are diphtheria toxoid, tetanus toxoid, inactivated pertussis toxin, hepatitis B virus surface antigen, inactivated Type I poliovirus (Mahoney), Type 2 poliovirus (MEF-1), Type 3 poliovirus (Saukett), filamentous hemagglutinin and pertactin (69 kD outer membrane protein), or analogs thereof. In one embodiment, the pharmaceutical composition is PEDIARIX® (Diphtheria and Tetanus Toxoids and Acellular Pertussis Adsorbed, Hepatitis B (Recombinant) and Inactivated Poliovirus Vaccine).
In one embodiment of any of the foregoing aspects of the invention, the active pharmaceutical ingredient is inactivated hepatitis A virus, or an analog thereof. In a particular embodiment, the pharmaceutical composition is HAVRIX® (Hepatitis A Vaccine).
In one embodiment of any of the foregoing aspects of the invention, the active pharmaceutical ingredient is noninfectious hepatitis B virus surface antigen (HBsAg), or an analog thereof. In a particular embodiment, the pharmaceutical composition is ENGERIX-B® (Hepatitis B Vaccine (Recombinant)).
In one embodiment of any of the foregoing aspects of the invention, the active pharmaceutical ingredients are inactivated hepatitis A virus and noninfectious hepatitis B virus surface antigen (HBsAg), or analogs thereof. In a particular embodiment, the pharmaceutical composition is TWINRIX® (Hepatitis A & Hepatitis B (Recombinant) Vaccine).
In one embodiment of any of the foregoing aspects of the invention, the active pharmaceutical ingredient is a glucagon-like peptide-1 (GLP-1) receptor agonist. In a particular embodiment, the pharmaceutical composition is EPERZAN® (albiglutide).
In one embodiment of any of the foregoing aspects of the invention, the active pharmaceutical ingredient is a melanoma associated antigen 3 (MAGE-A3) epitope fusion protein. In a particular embodiment, the pharmaceutical composition is MAGE-A3 Antigen-Specific Cancer Immunotherapeutic (astuprotimut-R).
In one embodiment of any of the foregoing aspects of the invention, the active pharmaceutical ingredient is a dystrophin antisense oligonucleotide of SEQ ID NO. 1. In a particular embodiment, the pharmaceutical composition is GSK2402968 (drisapersen).
In one embodiment of any of the foregoing aspects of the invention, the active pharmaceutical ingredient is a recombinant varicella zoster virus glycoprotein E. In a particular embodiment, the pharmaceutical composition is HZ/su (herpes zoster vaccine).
In a particular aspect, the present invention provides a delamination resistant pharmaceutical container formed, at least in part, of a glass composition including about 76.8 mol. % SiO2; about 6.0 mol. % Al2O3; about 11.6 mol. % Na2O; about 0.1 mol. % K2O; about 4.8 mol. % MgO; and about 0.5 mol. % CaO, wherein the glass composition is free of boron and compounds of boron; and wherein the pharmaceutical container further comprises a pharmaceutical composition selected from the group consisting of PEDIARIX® (Diphtheria and Tetanus Toxoids and Acellular Pertussis Adsorbed, Hepatitis B (Recombinant) and Inactivated Poliovirus Vaccine), HAVRIX® (Hepatitis A Vaccine), ENGERIX-B® (Hepatitis B Vaccine (Recombinant)), TWINRIX® (Hepatitis A & Hepatitis B (Recombinant) Vaccine), EPERZAN® (albiglutide), MAGE-A3 Antigen-Specific Cancer Immunotherapeutic (astuprotimut-R), GSK2402968 (drisapersen), and HZ/su (herpes zoster vaccine).
In one aspect, the present invention includes a delamination resistant pharmaceutical container including a glass composition. The pharmaceutical container includes from about 70 mol. % to about 80 mol. % SiO2; from about 3 mol. % to about 13 mol. % alkaline earth oxide; X mol. % Al2O3; and Y mol. % alkali oxide. The alkali oxide includes Na2O in an amount greater than about 8 mol. %, a ratio of Y:X is greater than 1, and the glass composition is free of boron and compounds of boron. The delamination resistant pharmaceutical container further includes an active pharmaceutical ingredient.
In one or more embodiments, the SiO2 is present in an amount less than or equal to 78 mol. %. In some embodiments, an amount of the alkaline earth oxide is greater than or equal to about 4 mol. % and less than or equal to about 8 mol. %. In one or more embodiments, the alkaline earth oxide includes MgO and CaO and a ratio (CaO (mol. %)/(CaO (mol. %)+MgO (mol. %))) is less than or equal to 0.5. In one or more embodiments, the alkaline earth oxide includes from about 0.1 mol. % to less than or equal to about 1.0 mol. % CaO. In one or more embodiments, the alkaline earth oxide includes from about 3 mol. % to about 7 mol. % MgO. In one or more embodiments, X is greater than or equal to about 2 mol. % and less than or equal to about 10 mol. %. In embodiments, the alkali oxide includes greater than or equal to about 9 mol. % Na2O and less than or equal to about 15 mol. % Na2O. In some embodiments, the ratio of Y:X is less than or equal to 2. In one or more embodiments, the ratio of Y:X is greater than or equal to 1.3 and less than or equal to 2.0. In one or more embodiments, the alkali oxide further includes K2O in an amount less than or equal to about 3 mol. %. In one or more embodiments, the glass composition is free of phosphorous and compounds of phosphorous. In one or more embodiments, the alkali oxide includes K2O in an amount greater than or equal to about 0.01 mol. % and less than or equal to about 1.0 mol. %.
In another aspect, the invention includes a delamination resistant pharmaceutical container including a pharmaceutical composition. The pharmaceutical container includes an active pharmaceutical ingredient, such that the pharmaceutical container includes a glass composition including SiO2 in a concentration greater than about 70 mol. %; alkaline earth oxide including MgO and CaO, wherein CaO is present in an amount greater than or equal to about 0.1 mol. % and less than or equal to about 1.0 mol. %, and a ratio (CaO (mol. %)/(CaO (mol. %)+MgO (mol. %))) is less than or equal to 0.5; and Y mol. % alkali oxide, wherein the alkali oxide includes Na2O in an amount greater than about 8 mol. %, such that the glass composition is free of boron and compounds of boron.
In another aspect, the invention includes a delamination resistant pharmaceutical container including a pharmaceutical composition including an active pharmaceutical ingredient. The pharmaceutical container includes a glass composition including from about 72 mol. % to about 78 mol. % SiO2; from about 4 mol. % to about 8 mol. % alkaline earth oxide, wherein the alkaline earth oxide includes MgO and CaO and a ratio (CaO (mol. %)/(CaO (mol. %)+MgO (mol. %))) is less than or equal to 0.5; X mol. % Al2O3, such that X is greater than or equal to about 4 mol. % and less than or equal to about 8 mol. %; and Y mol. % alkali oxide, such that the alkali oxide includes Na2O in an amount greater than or equal to about 9 mol. % and less than or equal to about 15 mol. %, a ratio of Y:X is greater than 1, and the glass composition is free of boron and compounds of boron.
In another aspect, the invention includes a delamination resistant pharmaceutical container including a pharmaceutical composition including an active pharmaceutical ingredient. The pharmaceutical container includes a glass composition. The glass composition includes from about 70 mol. % to about 80 mol. % SiO2; from about 3 mol. % to about 13 mol. % alkaline earth oxide, such that the alkaline earth oxide includes CaO in an amount greater than or equal to about 0.1 mol. % and less than or equal to about 1.0 mol. %, MgO, and a ratio (CaO (mol. %)/(CaO (mol. %)+MgO (mol. %))) is less than or equal to 0.5; X mol. % Al2O3, wherein X is greater than or equal to about 2 mol. % and less than or equal to about 10 mol. %; and Y mol. % alkali oxide, wherein the alkali oxide includes from about 0.01 mol. % to about 1.0 mol. % K2O and a ratio of Y:X is greater than 1, and the glass composition is free of boron and compounds of boron.
In one or more embodiments of any of the above aspects, the pharmaceutical composition includes PEDIARIX® (Diphtheria and Tetanus Toxoids and Acellular Pertussis Adsorbed, Hepatitis B (Recombinant) and Inactivated Poliovirus Vaccine), HAVRIX® (Hepatitis A Vaccine), ENGERIX-B® (Hepatitis B Vaccine (Recombinant)), TWINRIX® (Hepatitis A & Hepatitis B (Recombinant) Vaccine), EPERZAN® (albiglutide), MAGE-A3 Antigen-Specific Cancer Immunotherapeutic (astuprotimut-R), GSK2402968 (drisapersen), or HZ/su (herpes zoster vaccine).
In one aspect, the present invention includes a pharmaceutical composition. The pharmaceutical composition includes PEDIARIX® (Diphtheria and Tetanus Toxoids and Acellular Pertussis Adsorbed, Hepatitis B (Recombinant) and Inactivated Poliovirus Vaccine), HAVRIX® (Hepatitis A Vaccine), ENGERIX-B® (Hepatitis B Vaccine (Recombinant)), TWINRIX® (Hepatitis A & Hepatitis B (Recombinant) Vaccine), EPERZAN® (albiglutide), MAGE-A3 Antigen-Specific Cancer Immunotherapeutic (astuprotimut-R), GSK2402968 (drisapersen), or HZ/su (herpes zoster vaccine) and a pharmaceutically acceptable excipient, such that the pharmaceutical composition is contained within a glass pharmaceutical container including an internal homogeneous layer.
In one or more embodiments, the pharmaceutical container has a compressive stress greater than or equal to 150 MPa. In one or more embodiments, the pharmaceutical container has a compressive stress greater than or equal to 250 MPa. In one or more embodiments, the pharmaceutical container includes a depth of layer greater than 30 μm. In one or more embodiments, the depth of layer is greater than 35 μm. In one or more embodiments, the pharmaceutical composition demonstrates increased stability, product integrity, or efficacy.
In one aspect, the present invention includes a pharmaceutical composition. The pharmaceutical composition includes PEDIARIX® (Diphtheria and Tetanus Toxoids and Acellular Pertussis Adsorbed, Hepatitis B (Recombinant) and Inactivated Poliovirus Vaccine), HAVRIX® (Hepatitis A Vaccine), ENGERIX-B® (Hepatitis B Vaccine (Recombinant)), TWINRIX® (Hepatitis A & Hepatitis B (Recombinant) Vaccine), EPERZAN® (albiglutide), MAGE-A3 Antigen-Specific Cancer Immunotherapeutic (astuprotimut-R), GSK2402968 (drisapersen), or HZ/su (herpes zoster vaccine) and a pharmaceutically acceptable excipient, such that the pharmaceutical composition is contained within a glass pharmaceutical container including an internal homogeneous layer having a compressive stress greater than or equal to 150 MPa.
In one or more embodiments, the pharmaceutical container includes a depth of layer greater than 10 μm. In one or more embodiments, the pharmaceutical container includes a depth of layer greater than 25 μm. In one or more embodiments, the pharmaceutical container includes a depth of layer greater than 30 μm. In one or more embodiments, the pharmaceutical container has compressive stress greater than or equal to 300 MPa. In one or more embodiments, the pharmaceutical container includes increased stability, product integrity, or efficacy.
In another aspect, the present technology includes a pharmaceutical composition. The pharmaceutical composition includes PEDIARIX® (Diphtheria and Tetanus Toxoids and Acellular Pertussis Adsorbed, Hepatitis B (Recombinant) and Inactivated Poliovirus Vaccine), HAVRIX® (Hepatitis A Vaccine), ENGERIX-B® (Hepatitis B Vaccine (Recombinant)), TWINRIX® (Hepatitis A & Hepatitis B (Recombinant) Vaccine), EPERZAN® (albiglutide), MAGE-A3 Antigen-Specific Cancer Immunotherapeutic (astuprotimut-R), GSK2402968 (drisapersen), or HZ/su (herpes zoster vaccine) and a pharmaceutically acceptable excipient, such that the pharmaceutical composition is contained within a glass pharmaceutical container having a compressive stress greater than or equal to 150 MPa and a depth of layer greater than 10 μm, and such that the pharmaceutical composition demonstrates increased stability, product integrity, or efficacy.
In another aspect, the present technology includes a pharmaceutical composition. The pharmaceutical composition includes PEDIARIX® (Diphtheria and Tetanus Toxoids and Acellular Pertussis Adsorbed, Hepatitis B (Recombinant) and Inactivated Poliovirus Vaccine), HAVRIX® (Hepatitis A Vaccine), ENGERIX-B® (Hepatitis B Vaccine (Recombinant)), TWINRIX® (Hepatitis A & Hepatitis B (Recombinant) Vaccine), EPERZAN® (albiglutide), MAGE-A3 Antigen-Specific Cancer Immunotherapeutic (astuprotimut-R), GSK2402968 (drisapersen), or HZ/su (herpes zoster vaccine) and a pharmaceutically acceptable excipient, such that the pharmaceutical composition is contained within a glass pharmaceutical container including a substantially homogeneous inner layer, and such that the pharmaceutical composition demonstrates increased stability, product integrity, or efficacy.
In another aspect, the present technology includes a pharmaceutical composition. The pharmaceutical composition includes PEDIARIX® (Diphtheria and Tetanus Toxoids and Acellular Pertussis Adsorbed, Hepatitis B (Recombinant) and Inactivated Poliovirus Vaccine), HAVRIX® (Hepatitis A Vaccine), ENGERIX-B® (Hepatitis B Vaccine (Recombinant)), TWINRIX® (Hepatitis A & Hepatitis B (Recombinant) Vaccine), EPERZAN® (albiglutide), MAGE-A3 Antigen-Specific Cancer Immunotherapeutic (astuprotimut-R), GSK2402968 (drisapersen), or HZ/su (herpes zoster vaccine) and a pharmaceutically acceptable excipient, such that the pharmaceutical composition is contained within a glass pharmaceutical container having a delamination factor of less than 3, wherein the pharmaceutical composition demonstrates increased stability, product integrity, or efficacy.
In another aspect, the present technology includes a pharmaceutical composition. The pharmaceutical composition includes PEDIARIX® (Diphtheria and Tetanus Toxoids and Acellular Pertussis Adsorbed, Hepatitis B (Recombinant) and Inactivated Poliovirus Vaccine), HAVRIX® (Hepatitis A Vaccine), ENGERIX-B® (Hepatitis B Vaccine (Recombinant)), TWINRIX® (Hepatitis A & Hepatitis B (Recombinant) Vaccine), EPERZAN® (albiglutide), MAGE-A3 Antigen-Specific Cancer Immunotherapeutic (astuprotimut-R), GSK2402968 (drisapersen), or HZ/su (herpes zoster vaccine) and a pharmaceutically acceptable excipient, such that the pharmaceutical composition is contained within a glass pharmaceutical container which is substantially free of boron, and such that the pharmaceutical composition demonstrates increased stability, product integrity, or efficacy.
In one or more embodiments, the glass pharmaceutical container has a compressive stress greater than or equal to 150 MPa and a depth of layer greater than 25 μm. In one or more embodiments, the glass pharmaceutical container has a compressive stress greater than or equal to 300 MPa and a depth of layer greater than 35 μm. In one or more embodiments, the glass pharmaceutical container includes a substantially homogeneous inner layer. In one or more embodiments, the glass pharmaceutical container has a compressive stress greater than or equal to 150 MPa and a depth of layer greater than 25 μm.
In another aspect, the present technology includes a pharmaceutical composition. The pharmaceutical composition includes PEDIARIX® (Diphtheria and Tetanus Toxoids and Acellular Pertussis Adsorbed, Hepatitis B (Recombinant) and Inactivated Poliovirus Vaccine), HAVRIX® (Hepatitis A Vaccine), ENGERIX-B® (Hepatitis B Vaccine (Recombinant)), TWINRIX® (Hepatitis A & Hepatitis B (Recombinant) Vaccine), EPERZAN® (albiglutide), MAGE-A3 Antigen-Specific Cancer Immunotherapeutic (astuprotimut-R), GSK2402968 (drisapersen), or HZ/su (herpes zoster vaccine) and a pharmaceutically acceptable excipient, such that the pharmaceutical composition is contained within a glass pharmaceutical container including a delamination factor of less than 3, and such that the pharmaceutical composition includes increased stability, product integrity, or efficacy.
In one or more embodiments of any of the above aspects, the container has a compressive stress greater than or equal to 300 MPa. In one or more embodiments, the container has a depth of layer greater than 25 μm. In one or more embodiments, the container has a depth of layer greater than 30 μm. In one or more embodiments, the container has a depth of layer of at least 35 μm. In one or more embodiments, the container has a compressive stress greater than or equal to 300 MPa. In one or more embodiments, the container has a compressive stress greater than or equal to 350 MPa.
Additional features and advantages will be set forth in the detailed description which follows, and in part will be readily apparent to those skilled in the art from that description or recognized by practicing the embodiments described herein, including the detailed description which follows, the claims, as well as the appended drawings.
It is to be understood that both the foregoing general description and the following detailed description describe various embodiments and are intended to provide an overview or framework for understanding the nature and character of the claimed subject matter. The accompanying drawings are included to provide a further understanding of the various embodiments, and are incorporated into and constitute a part of this specification. The drawings illustrate the various embodiments described herein, and together with the description serve to explain the principles and operations of the claimed subject matter.



BRIEF DESCRIPTION OF THE DRAWINGS
FIG. 1 graphically depicts the relationship between the ratio of alkali oxides to alumina (x-axis) and the strain point, annealing point, and softening point (y-axes) of inventive and comparative glass compositions;
FIG. 2 graphically depicts the relationship between the ratio of alkali oxides to alumina (x-axis) and the maximum compressive stress and stress change (y-axes) of inventive and comparative glass compositions;
FIG. 3 graphically depicts the relationship between the ratio of alkali oxides to alumina (x-axis) and hydrolytic resistance as determined from the ISO 720 standard (y-axis) of inventive and comparative glass compositions;
FIG. 4 graphically depicts diffusivity D (y-axis) as a function of the ratio (CaO/(CaO+MgO)) (x-axis) for inventive and comparative glass compositions;
FIG. 5 graphically depicts the maximum compressive stress (y-axis) as a function of the ratio (CaO/(CaO+MgO)) (x-axis) for inventive and comparative glass compositions;
FIG. 6 graphically depicts diffusivity D (y-axis) as a function of the ratio (B2O3/(R2O−Al2O3)) (x-axis) for inventive and comparative glass compositions; and
FIG. 7 graphically depicts the hydrolytic resistance as determined from the ISO 720 standard (y-axis) as a function of the ratio (B2O3/(R2O−Al2O3)) (x-axis) for inventive and comparative glass compositions.



DETAILED DESCRIPTION
The present invention is based, at least in part, on the identification of a pharmaceutical container formed, at least in part, of a glass composition which exhibits a reduced propensity to delaminate, i.e., a reduced propensity to shed glass particulates. As a result, the presently claimed containers are particularly suited for storage, maintenance and/or delivery of therapeutically efficacious pharmaceutical compositions and, in particular pharmaceutical solutions comprising active pharmaceutical ingredients, for example, PEDIARIX® (Diphtheria and Tetanus Toxoids and Acellular Pertussis Adsorbed, Hepatitis B (Recombinant) and Inactivated Poliovirus Vaccine), HAVRIX® (Hepatitis A Vaccine), ENGERIX-B® (Hepatitis B Vaccine (Recombinant)), TWINRIX® (Hepatitis A & Hepatitis B (Recombinant) Vaccine), EPERZAN® (albiglutide), MAGE-A3 Antigen-Specific Cancer Immunotherapeutic (astuprotimut-R), GSK2402968 (drisapersen), and HZ/su (herpes zoster vaccine).
Conventional glass containers or glass packages for containing pharmaceutical compositions are generally formed from glass compositions which are known to exhibit chemical durability and low thermal expansion, such as alkali borosilicate glasses. While alkali borosilicate glasses exhibit good chemical durability, container manufacturers have sporadically observed silica-rich glass flakes dispersed in the solution contained in the glass containers as a result of delamination, particularly when the solution has been stored in direct contact with the glass surface for long time periods (months to years).
Delamination refers to a phenomenon in which glass particles are released from the surface of the glass following a series of leaching, corrosion, and/or weathering reactions. In general, the glass particles are silica-rich flakes of glass which originate from the interior surface of the package as a result of the leaching of modifier ions into a solution contained within the package. These flakes may generally be from about 1 nm to 2 μm thick with a width greater than about 50 μm.
It has heretofore been hypothesized that delamination is due to the phase separation which occurs in alkali borosilicate glasses when the glass is exposed to the elevated temperatures used for reforming the glass into a container shape.
However, it is now believed that the delamination of the silica-rich glass flakes from the interior surfaces of the glass containers is due to the compositional characteristics of the glass container in its as-formed condition. Specifically, the high silica content of alkali borosilicate glasses increases the melting temperature of the glass. However, the alkali and borate components in the glass composition melt and/or vaporize at much lower temperatures. In particular, the borate species in the glass are highly volatile and evaporate from the surface of the glass at the high temperatures necessary to melt and form the glass.
Specifically, glass stock is reformed into glass containers at high temperatures and in direct flames. The high temperatures cause the volatile borate species to evaporate from portions of the surface of the glass. When this evaporation occurs within the interior volume of the glass container, the volatilized borate species are re-deposited in other areas of the glass causing compositional heterogeneities in the glass container, particularly with respect to the bulk of the glass container. For example, as one end of a glass tube is closed to form the bottom or floor of the container, borate species may evaporate from the bottom portion of the tube and be re-deposited elsewhere in the tube. As a result, the areas of the container exposed to higher temperatures have silica-rich surfaces. Other areas of the container which are amenable to boron deposition may have a silica-rich surface with a boron-rich layer below the surface. Areas amenable to boron deposition are at a temperature greater than the anneal point of the glass composition but less than the hottest temperature the glass is subjected to during reformation when the boron is incorporated into the surface of the glass. Solutions contained in the container may leach the boron from the boron-rich layer. As the boron-rich layer is leached from the glass, the silica-rich surface begins to spall, shedding silica-rich flakes into the solution.
Definitions
The term “softening point,” as used herein, refers to the temperature at which the viscosity of the glass composition is 1×107.6 poise.
The term “annealing point,” as used herein, refers to the temperature at which the viscosity of the glass composition is 1×1013 poise.
The terms “strain point” and “Tstrain” as used herein, refers to the temperature at which the viscosity of the glass composition is 3×1014 poise.
The term “CTE,” as used herein, refers to the coefficient of thermal expansion of the glass composition over a temperature range from about room temperature (RT) to about 300° C.
In the embodiments of the glass compositions described herein, the concentrations of constituent components (e.g., SiO2, Al2O3, and the like) are specified in mole percent (mol. %) on an oxide basis, unless otherwise specified.
The terms “free” and “substantially free,” when used to describe the concentration and/or absence of a particular constituent component in a glass composition, means that the constituent component is not intentionally added to the glass composition. However, the glass composition may contain traces of the constituent component as a contaminant or tramp in amounts of less than 0.01 mol. %.
The term “chemical durability,” as used herein, refers to the ability of the glass composition to resist degradation upon exposure to specified chemical conditions. Specifically, the chemical durability of the glass compositions described herein was assessed according to three established material testing standards: DIN 12116 dated March 2001 and entitled “Testing of glass—Resistance to attack by a boiling aqueous solution of hydrochloric acid—Method of test and classification”; ISO 695:1991 entitled “Glass—Resistance to attack by a boiling aqueous solution of mixed alkali—Method of test and classification”; and ISO 720:1985 entitled “Glass—Hydrolytic resistance of glass grains at 121 degrees C.—Method of test and classification.” The chemical durability of the glass may also be assessed according to ISO 719:1985 “Glass—Hydrolytic resistance of glass grains at 98 degrees C.—Method of test and classification,” in addition to the above referenced standards. The ISO 719 standard is a less rigorous version of the ISO 720 standard and, as such, it is believed that a glass which meets a specified classification of the ISO 720 standard will also meet the corresponding classification of the ISO 719 standard. The classifications associated with each standard are described in further detail herein.
Glass Compositions
Reference will now be made in detail to various embodiments of pharmaceutical containers formed, at least in part, of glass compositions which exhibit improved chemical and mechanical durability and, in particular, improved resistance to delamination. The glass compositions may also be chemically strengthened thereby imparting increased mechanical durability to the glass. The glass compositions described herein generally comprise silica (SiO2), alumina (Al2O3), alkaline earth oxides (such as MgO and/or CaO), and alkali oxides (such as Na2O and/or K2O) in amounts which impart chemical durability to the glass composition. Moreover, the alkali oxides present in the glass compositions facilitate chemically strengthening the glass compositions by ion exchange. Various embodiments of the glass compositions will be described herein and further illustrated with reference to specific examples.
The glass compositions described herein are alkali aluminosilicate glass compositions which generally include a combination of SiO2, Al2O3, at least one alkaline earth oxide, and one or more alkali oxides, such as Na2O and/or K2O. In some embodiments, the glass compositions may be free from boron and compounds containing boron. The combination of these components enables a glass composition which is resistant to chemical degradation and is also suitable for chemical strengthening by ion exchange. In some embodiments the glass compositions may further comprise minor amounts of one or more additional oxides such as, for example, SnO2, ZrO2, ZnO, TiO2, As2O3 or the like. These components may be added as fining agents and/or to further enhance the chemical durability of the glass composition.
In the embodiments of the glass compositions described herein SiO2 is the largest constituent of the composition and, as such, is the primary constituent of the resulting glass network. SiO2 enhances the chemical durability of the glass and, in particular, the resistance of the glass composition to decomposition in acid and the resistance of the glass composition to decomposition in water. Accordingly, a high SiO2 concentration is generally desired. However, if the content of SiO2 is too high, the formability of the glass may be diminished as higher concentrations of SiO2 increase the difficulty of melting the glass which, in turn, adversely impacts the formability of the glass. In the embodiments described herein, the glass composition generally comprises SiO2 in an amount greater than or equal to 67 mol. % and less than or equal to about 80 mol. % or even less than or equal to 78 mol. %. In some embodiments, the amount of SiO2 in the glass composition may be greater than about 68 mol. %, greater than about 69 mol. % or even greater than about 70 mol. %. In some other embodiments, the amount of SiO2 in the glass composition may be greater than 72 mol. %, greater than 73 mol. % or even greater than 74 mol. %. For example, in some embodiments, the glass composition may include from about 68 mol. % to about 80 mol. % or even to about 78 mol. % SiO2. In some other embodiments the glass composition may include from about 69 mol. % to about 80 mol. % or even to about 78 mol. % SiO2. In some other embodiments the glass composition may include from about 70 mol. % to about 80 mol. % or even to about 78 mol. % SiO2. In still other embodiments, the glass composition comprises SiO2 in an amount greater than or equal to 70 mol. % and less than or equal to 78 mol. %. In some embodiments, SiO2 may be present in the glass composition in an amount from about 72 mol. % to about 78 mol. %. In some other embodiments, SiO2 may be present in the glass composition in an amount from about 73 mol. % to about 78 mol. %. In other embodiments, SiO2 may be present in the glass composition in an amount from about 74 mol. % to about 78 mol. %. In still other embodiments, SiO2 may be present in the glass composition in an amount from about 70 mol. % to about 76 mol. %.
The glass compositions described herein further include Al2O3. Al2O3, in conjunction with alkali oxides present in the glass compositions such as Na2O or the like, improves the susceptibility of the glass to ion exchange strengthening. In the embodiments described herein, Al2O3 is present in the glass compositions in X mol. % while the alkali oxides are present in the glass composition in Y mol. %. The ratio Y:X in the glass compositions described herein is greater than 1 in order to facilitate the aforementioned susceptibility to ion exchange strengthening. Specifically, the diffusion coefficient or diffusivity D of the glass composition relates to the rate at which alkali ions penetrate into the glass surface during ion exchange. Glasses which have a ratio Y:X greater than about 0.9 or even greater than about 1 have a greater diffusivity than glasses which have a ratio Y:X less than 0.9. Glasses in which the alkali ions have a greater diffusivity can obtain a greater depth of layer for a given ion exchange time and ion exchange temperature than glasses in which the alkali ions have a lower diffusivity. Moreover, as the ratio of Y:X increases, the strain point, anneal point, and softening point of the glass decrease, such that the glass is more readily formable. In addition, for a given ion exchange time and ion exchange temperature, it has been found that compressive stresses induced in glasses which have a ratio Y:X greater than about 0.9 and less than or equal to 2 are generally greater than those generated in glasses in which the ratio Y:X is less than 0.9 or greater than 2. Accordingly, in some embodiments, the ratio of Y:X is greater than 0.9 or even greater than 1. In some embodiments, the ratio of Y:X is greater than 0.9, or even greater than 1, and less than or equal to about 2. In still other embodiments, the ratio of Y:X may be greater than or equal to about 1.3 and less than or equal to about 2.0 in order to maximize the amount of compressive stress induced in the glass for a specified ion exchange time and a specified ion exchange temperature.
However, if the amount of Al2O3 in the glass composition is too high, the resistance of the glass composition to acid attack is diminished. Accordingly, the glass compositions described herein generally include Al2O3 in an amount greater than or equal to about 2 mol. % and less than or equal to about 10 mol. %. In some embodiments, the amount of Al2O3 in the glass composition is greater than or equal to about 4 mol. % and less than or equal to about 8 mol. %. In some other embodiments, the amount of Al2O3 in the glass composition is greater than or equal to about 5 mol. % to less than or equal to about 7 mol. %. In some other embodiments, the amount of Al2O3 in the glass composition is greater than or equal to about 6 mol. % to less than or equal to about 8 mol. %. In still other embodiments, the amount of Al2O3 in the glass composition is greater than or equal to about 5 mol. % to less than or equal to about 6 mol. %.
The glass compositions also include one or more alkali oxides such as Na2O and/or K2O. The alkali oxides facilitate the ion exchangeability of the glass composition and, as such, facilitate chemically strengthening the glass. The alkali oxide may include one or more of Na2O and K2O. The alkali oxides are generally present in the glass composition in a total concentration of Y mol. %. In some embodiments described herein, Y may be greater than about 2 mol. % and less than or equal to about 18 mol. %. In some other embodiments, Y may be greater than about 8 mol. %, greater than about 9 mol. %, greater than about 10 mol. % or even greater than about 11 mol. %. For example, in some embodiments described herein Y is greater than or equal to about 8 mol. % and less than or equal to about 18 mol. %. In still other embodiments, Y may be greater than or equal to about 9 mol. % and less than or equal to about 14 mol. %.
The ion exchangeability of the glass composition is primarily imparted to the glass composition by the amount of the alkali oxide Na2O initially present in the glass composition prior to ion exchange. Accordingly, in the embodiments of the glass compositions described herein, the alkali oxide present in the glass composition includes at least Na2O. Specifically, in order to achieve the desired compressive stress and depth of layer in the glass composition upon ion exchange strengthening, the glass compositions include Na2O in an amount from about 2 mol. % to about 15 mol. % based on the molecular weight of the glass composition. In some embodiments the glass composition includes at least about 8 mol. % of Na2O based on the molecular weight of the glass composition. For example, the concentration of Na2O may be greater than 9 mol. %, greater than 10 mol. % or even greater than 11 mol. %. In some embodiments, the concentration of Na2O may be greater than or equal to 9 mol. % or even greater than or equal to 10 mol. %. For example, in some embodiments the glass composition may include Na2O in an amount greater than or equal to about 9 mol. % and less than or equal to about 15 mol. % or even greater than or equal to about 9 mol. % and less than or equal to 13 mol. %.
As noted above, the alkali oxide in the glass composition may further include K2O. The amount of K2O present in the glass composition also relates to the ion exchangeability of the glass composition. Specifically, as the amount of K2O present in the glass composition increases, the compressive stress obtainable through ion exchange decreases as a result of the exchange of potassium and sodium ions. Accordingly, it is desirable to limit the amount of K2O present in the glass composition. In some embodiments, the amount of K2O is greater than or equal to 0 mol. % and less than or equal to 3 mol. %. In some embodiments, the amount of K2O is less or equal to 2 mol. % or even less than or equal to 1.0 mol. %. In embodiments where the glass composition includes K2O, the K2O may be present in a concentration greater than or equal to about 0.01 mol. % and less than or equal to about 3.0 mol. % or even greater than or equal to about 0.01 mol. % and less than or equal to about 2.0 mol. %. In some embodiments, the amount of K2O present in the glass composition is greater than or equal to about 0.01 mol. % and less than or equal to about 1.0 mol. %. Accordingly, it should be understood that K2O need not be present in the glass composition. However, when K2O is included in the glass composition, the amount of K2O is generally less than about 3 mol. % based on the molecular weight of the glass composition.
The alkaline earth oxides present in the composition improve the meltability of the glass batch materials and increase the chemical durability of the glass composition. In the glass compositions described herein, the total mol. % of alkaline earth oxides present in the glass compositions is generally less than the total mol. % of alkali oxides present in the glass compositions in order to improve the ion exchangeability of the glass composition. In the embodiments described herein, the glass compositions generally include from about 3 mol. % to about 13 mol. % of alkaline earth oxide. In some of these embodiments, the amount of alkaline earth oxide in the glass composition may be from about 4 mol. % to about 8 mol. % or even from about 4 mol. % to about 7 mol. %.
The alkaline earth oxide in the glass composition may include MgO, CaO, SrO, BaO or combinations thereof. In some embodiments, the alkaline earth oxide includes MgO, CaO or combinations thereof. For example, in the embodiments described herein the alkaline earth oxide includes MgO. MgO is present in the glass composition in an amount which is greater than or equal to about 3 mol. % and less than or equal to about 8 mol. % MgO. In some embodiments, MgO may be present in the glass composition in an amount which is greater than or equal to about 3 mol. % and less than or equal to about 7 mol. % or even greater than or equal to 4 mol. % and less than or equal to about 7 mol. % by molecular weight of the glass composition.
In some embodiments, the alkaline earth oxide may further include CaO. In these embodiments CaO is present in the glass composition in an amount from about 0 mol. % to less than or equal to 6 mol. % by molecular weight of the glass composition. For example, the amount of CaO present in the glass composition may be less than or equal to 5 mol. %, less than or equal to 4 mol. %, less than or equal to 3 mol. %, or even less than or equal to 2 mol. %. In some of these embodiments, CaO may be present in the glass composition in an amount greater than or equal to about 0.1 mol. % and less than or equal to about 1.0 mol. %. For example, CaO may be present in the glass composition in an amount greater than or equal to about 0.2 mol. % and less than or equal to about 0.7 mol. % or even in an amount greater than or equal to about 0.3 mol. % and less than or equal to about 0.6 mol. %.
In the embodiments described herein, the glass compositions are generally rich in MgO, (i.e., the concentration of MgO in the glass composition is greater than the concentration of the other alkaline earth oxides in the glass composition including, without limitation, CaO). Forming the glass composition such that the glass composition is MgO-rich improves the hydrolytic resistance of the resultant glass, particularly following ion exchange strengthening. Moreover, glass compositions which are MgO-rich generally exhibit improved ion exchange performance relative to glass compositions which are rich in other alkaline earth oxides. Specifically, glasses formed from MgO-rich glass compositions generally have a greater diffusivity than glass compositions which are rich in other alkaline earth oxides, such as CaO. The greater diffusivity enables the formation of a deeper depth of layer in the glass. MgO-rich glass compositions also enable a higher compressive stress to be achieved in the surface of the glass compared to glass compositions which are rich in other alkaline earth oxides such as CaO. In addition, it is generally understood that as the ion exchange process proceeds and alkali ions penetrate more deeply into the glass, the maximum compressive stress achieved at the surface of the glass may decrease with time. However, glasses formed from glass compositions which are MgO-rich exhibit a lower reduction in compressive stress than glasses formed from glass compositions that are CaO-rich or rich in other alkaline earth oxides (i.e., glasses which are MgO-poor). Thus, MgO-rich glass compositions enable glasses which have higher compressive stress at the surface and greater depths of layer than glasses which are rich in other alkaline earth oxides.
In order to fully realize the benefits of MgO in the glass compositions described herein, it has been determined that the ratio of the concentration of CaO to the sum of the concentration of CaO and the concentration of MgO in mol. % (i.e., (CaO/(CaO+MgO)) should be minimized. Specifically, it has been determined that (CaO/(CaO+MgO)) should be less than or equal to 0.5. In some embodiments (CaO/(CaO+MgO)) is less than or equal to 0.3 or even less than or equal to 0.2. In some other embodiments (CaO/(CaO+MgO)) may even be less than or equal to 0.1.
Boron oxide (B2O3) is a flux which may be added to glass compositions to reduce the viscosity at a given temperature (e.g., the strain, anneal and softening temperatures) thereby improving the formability of the glass. However, it has been found that additions of boron significantly decrease the diffusivity of sodium and potassium ions in the glass composition which, in turn, adversely impacts the ion exchange performance of the resultant glass. In particular, it has been found that additions of boron significantly increase the time required to achieve a given depth of layer relative to glass compositions which are boron free. Accordingly, in some embodiments described herein, the amount of boron added to the glass composition is minimized in order to improve the ion exchange performance of the glass composition.
For example, it has been determined that the impact of boron on the ion exchange performance of a glass composition can be mitigated by controlling the ratio of the concentration of B2O3 to the difference between the total concentration of the alkali oxides (i.e., R2O, where R is the alkali metals) and alumina (i.e., B2O3 (mol. %)/(R2O (mol. %)−Al2O3 (mol. %)). In particular, it has been determined that when the ratio of B2O3/(R2O−Al2O3) is greater than or equal to about 0 and less than about 0.3 or even less than about 0.2, the diffusivities of alkali oxides in the glass compositions are not diminished and, as such, the ion exchange performance of the glass composition is maintained. Accordingly, in some embodiments, the ratio of B2O3/(R2O−Al2O3) is greater than 0 and less than or equal to 0.3. In some of these embodiments, the ratio of B2O3/(R2O−Al2O3) is greater than 0 and less than or equal to 0.2. In some embodiments, the ratio of B2O3/(R2O−Al2O3) is greater than 0 and less than or equal to 0.15 or even less than or equal to 0.1. In some other embodiments, the ratio of B2O3/(R2O−Al2O3) may be greater than 0 and less than or equal to 0.05. Maintaining the ratio B2O3/(R2O−Al2O3) to be less than or equal to 0.3 or even less than or equal to 0.2 permits the inclusion of B2O3 to lower the strain point, anneal point and softening point of the glass composition without the B2O3 adversely impacting the ion exchange performance of the glass.
In the embodiments described herein, the concentration of B2O3 in the glass composition is generally less than or equal to about 4 mol. %, less than or equal to about 3 mol. %, less than or equal to about 2 mol. %, or even less than or equal to 1 mol. %. For example, in embodiments where B2O3 is present in the glass composition, the concentration of B2O3 may be greater than about 0.01 mol. % and less than or equal to 4 mol. %. In some of these embodiments, the concentration of B2O3 may be greater than about 0.01 mol. % and less than or equal to 3 mol. % In some embodiments, the B2O3 may be present in an amount greater than or equal to about 0.01 mol. % and less than or equal to 2 mol. %, or even less than or equal to 1.5 mol. %. Alternatively, the B2O3 may be present in an amount greater than or equal to about 1 mol. % and less than or equal to 4 mol. %, greater than or equal to about 1 mol. % and less than or equal to 3 mol. % or even greater than or equal to about 1 mol. % and less than or equal to 2 mol. %. In some of these embodiments, the concentration of B2O3 may be greater than or equal to about 0.1 mol. % and less than or equal to 1.0 mol. %.
While in some embodiments the concentration of B2O3 in the glass composition is minimized to improve the forming properties of the glass without detracting from the ion exchange performance of the glass, in some other embodiments the glass compositions are free from boron and compounds of boron such as B2O3. Specifically, it has been determined that forming the glass composition without boron or compounds of boron improves the ion exchangeability of the glass compositions by reducing the process time and/or temperature required to achieve a specific value of compressive stress and/or depth of layer.
In some embodiments of the glass compositions described herein, the glass compositions are free from phosphorous and compounds containing phosphorous including, without limitation, P2O5. Specifically, it has been determined that formulating the glass composition without phosphorous or compounds of phosphorous increases the chemical durability of the glass composition.
In addition to the SiO2, Al2O3, alkali oxides and alkaline earth oxides, the glass compositions described herein may optionally further comprise one or more fining agents such as, for example, SnO2, As2O3, and/or Cl− (from NaCl or the like). When a fining agent is present in the glass composition, the fining agent may be present in an amount less than or equal to about 1 mol. % or even less than or equal to about 0.4 mol. %. For example, in some embodiments the glass composition may include SnO2 as a fining agent. In these embodiments SnO2 may be present in the glass composition in an amount greater than about 0 mol. % and less than or equal to about 1 mol. % or even an amount greater than or equal to about 0.01 mol. % and less than or equal to about 0.30 mol. %.
Moreover, the glass compositions described herein may comprise one or more additional metal oxides to further improve the chemical durability of the glass composition. For example, the glass composition may further include ZnO, TiO2, or ZrO2, each of which further improves the resistance of the glass composition to chemical attack. In these embodiments, the additional metal oxide may be present in an amount which is greater than or equal to about 0 mol. % and less than or equal to about 2 mol. %. For example, when the additional metal oxide is ZnO, the ZnO may be present in an amount greater than or equal to 1 mol. % and less than or equal to about 2 mol. %. When the additional metal oxide is ZrO2 or TiO2, the ZrO2 or TiO2 may be present in an amount less than or equal to about 1 mol. %.
Based on the foregoing, it should be understood that, in a first exemplary embodiment, a glass composition may include: SiO2 in a concentration greater than about 70 mol. % and Y mol. % alkali oxide. The alkali oxide may include Na2O in an amount greater than about 8 mol. %. The glass composition may be free of boron and compounds of boron. The concentration of SiO2 in this glass composition may be greater than or equal to about 72 mol. %, greater than 73 mol. % or even greater than 74 mol. %. The glass composition of this first exemplary embodiment may be free from phosphorous and compounds of phosphorous. The glass composition may also include X mol. % Al2O3. When Al2O3 is included, the ratio of Y:X may be greater than 1. The concentration of Al2O3 may be greater than or equal to about 2 mol. % and less than or equal to about 10 mol. %.
The glass composition of this first exemplary embodiment may also include alkaline earth oxide in an amount from about 3 mol. % to about 13 mol. %. The alkaline earth oxide may include MgO and CaO. The CaO may be present in an amount greater than or equal to about 0.1 mol. % and less than or equal to about 1.0 mol. %. A ratio (CaO (mol. %)/(CaO (mol. %)+MgO (mol. %))) may be less than or equal to 0.5.
In a second exemplary embodiment, a glass composition may include: greater than about 68 mol. % SiO2; X mol. % Al2O3; Y mol. % alkali oxide; and B2O3. The alkali oxide may include Na2O in an amount greater than about 8 mol %. A ratio (B2O3 (mol. %)/(Y mol. %−X mol. %) may be greater than 0 and less than 0.3. The concentration of SiO2 in this glass composition may be greater than or equal to about 72 mol. %, greater than 73 mol. % or even greater than 74 mol. %. The concentration of Al2O3 may be greater than or equal to about 2 mol. % and less than or equal to about 10 mol. %. In this second exemplary embodiment, the ratio of Y:X may be greater than 1. When the ratio of Y:X is greater than 1, an upper bound of the ratio of Y:X may be less than or equal to 2. The glass composition of this first exemplary embodiment may be free from phosphorous and compounds of phosphorous.
The glass composition of this second exemplary embodiment may also include alkaline earth oxide. The alkaline earth oxide may include MgO and CaO. The CaO may be present in an amount greater than or equal to about 0.1 mol. % and less than or equal to about 1.0 mol. %. A ratio (CaO (mol. %)/(CaO (mol. %)+MgO (mol. %))) may be less than or equal to 0.5.
The concentration of B2O3 in this second exemplary embodiment may be greater than or equal to about 0.01 mol. % and less than or equal to about 4 mol. %.
In a third exemplary embodiment, a glass article may have a type HgB1 hydrolytic resistance according to ISO 719. The glass article may include greater than about 8 mol. % Na2O and less than about 4 mol. % B2O3. The glass article may further comprise X mol. % Al2O3 and Y mol. % alkali oxide. The ratio (B2O3 (mol. %)/(Y mol. %−X mol. %) may be greater than 0 and less than 0.3. The glass article of this third exemplary embodiment may further include a compressive stress layer having a surface compressive stress greater than or equal to about 250 MPa. The glass article may also have at least a class S3 acid resistance according to DIN 12116; at least a class A2 base resistance according to ISO 695; and a type HgA1 hydrolytic resistance according to ISO 720.
In a fourth exemplary embodiment, a glass pharmaceutical package may include SiO2 in an amount greater than about 70 mol. %; X mol. % Al2O3; and Y mol. % alkali oxide. The alkali oxide may include Na2O in an amount greater than about 8 mol. %. A ratio of a concentration of B2O3 (mol. %) in the glass pharmaceutical package to (Y mol. %−X mol. %) may be less than 0.3. The glass pharmaceutical package may also have a type HGB1 hydrolytic resistance according to ISO 719. The concentration of SiO2 in the glass pharmaceutical package of this fourth exemplary embodiment may be greater than or equal to 72 mol. % and less than or equal to about 78 mol. % or even greater than 74 mol. % and less than or equal to about 78 mol. %. The concentration of Al2O3 in the glass pharmaceutical may be greater than or equal to about 4 mol. % and less than or equal to about 8 mol. %. A ratio of Y:X may be greater than 1 and less than 2.
The glass pharmaceutical package of this fourth exemplary embodiment may also include alkaline earth oxide in an amount from about 4 mol. % to about 8 mol. %. The alkaline earth oxide may include MgO and CaO. The CaO may be present in an amount greater than or equal to about 0.2 mol. % and less than or equal to about 0.7 mol. %. A ratio (CaO (mol. %)/(CaO (mol. %)+MgO (mol. %))) may be less than or equal to 0.5. The glass pharmaceutical package of this fourth exemplary embodiment may have a type HGA1 hydrolytic resistance according to ISO 720.
In a fifth exemplary embodiment, a glass composition may include from about 70 mol. % to about 80 mol. % SiO2; from about 3 mol. % to about 13 mol. % alkaline earth oxide; X mol. % Al2O3; and Y mol. % alkali oxide. The alkali oxide may include Na2O in an amount greater than about 8 mol. %. A ratio of Y:X may be greater than 1. The glass composition may be free of boron and compounds of boron.
In a sixth exemplary embodiment, a glass composition may include from about 68 mol. % to about 80 mol. % SiO2; from about 3 mol. % to about 13 mol. % alkaline earth oxide; X mol. % Al2O3; and Y mol. % alkali oxide. The alkali oxide may include Na2O in an amount greater than about 8 mol. %. The glass composition of this sixth exemplary embodiment may also include B2O3. A ratio (B2O3 (mol. %)/(Y mol. %−X mol. %) may be greater than 0 and less than 0.3. A ratio of Y:X may be greater than 1.
In a seventh exemplary embodiment, a glass composition may include from about 70 mol. % to about 80 mol. % SiO2; from about 3 mol. % to about 13 mol. % alkaline earth oxide; X mol. % Al2O3; and Y mol. % alkali oxide. The amount of Al2O3 in the glass composition may be greater than or equal to about 2 mol. % and less than or equal to about 10 mol. %. The alkaline earth oxide may include CaO in an amount greater than or equal to about 0.1 mol. % and less than or equal to about 1.0 mol. %. The alkali oxide may include from about 0.01 mol. % to about 1.0 mol. % K2O. A ratio of Y:X may be greater than 1. The glass composition may be free of boron and compounds of boron. The glass composition may be amenable to strengthening by ion exchange.
In a seventh exemplary embodiment, a glass composition may include SiO2 in an amount greater than about 70 mol. % and less than or equal to about 80 mol. %; X mol. % Al2O3; and Y mol. % alkali oxide. The alkali oxide may include Na2O in an amount greater than about 8 mol. %. A ratio of a concentration of B2O3 (mol. %) in the glass pharmaceutical package to (Y mol. %−X mol. %) may be less than 0.3. A ratio of Y:X may be greater than 1.
In an eighth exemplary embodiment, a glass composition may include from about 72 mol. % to about 78 mol. % SiO2; from about 4 mol. % to about 8 mol. % alkaline earth oxide; X mol. % Al2O3, wherein X is greater than or equal to about 4 mol. % and less than or equal to about 8 mol. %; and Y mol. % alkali oxide, wherein the alkali oxide comprises Na2O in an amount greater than or equal to about 9 mol. % and less than or equal to about 15 mol. %. A ratio of a concentration of B2O3 (mol. %) in the glass pharmaceutical package to (Y mol. %−X mol. %) is less than 0.3. A ratio of Y:X may be greater than 1.
In a ninth exemplary embodiment, a pharmaceutical package for containing a pharmaceutical composition may include from about 70 mol. % to about 78 mol. % SiO2; from about 3 mol. % to about 13 mol. % alkaline earth oxide; X mol. % Al2O3, wherein X is greater than or equal to 2 mol. % and less than or equal to about 10 mol. %; and Y mol. % alkali oxide, wherein the alkali oxide comprises Na2O in an amount greater than about 8 mol. %. The alkaline earth oxide may include CaO in an amount less than or equal to about 6.0 mol. %. A ratio of Y:X may be greater than about 1. The package may be free of boron and compounds of boron and may include a compressive stress layer with a compressive stress greater than or equal to about 250 MPa and a depth of layer greater than or equal to about 10 μm.
In a tenth exemplary embodiment, a glass article may be formed from a glass composition comprising from about 70 mol. % to about 78 mol. % SiO2; alkaline earth oxide, wherein the alkaline earth oxide comprises MgO and CaO and a ratio (CaO (mol. %)/(CaO (mol. %)+MgO (mol. %))) is less than or equal to 0.5; X mol. % Al2O3, wherein X is from about 2 mol. % to about 10 mol. %; and Y mol. % alkali oxide, wherein the alkali oxide comprises Na2O in an amount greater than about 8 mol. % and a ratio of Y:X is greater than 1. The glass article may be ion exchange strengthened with a compressive stress greater than or equal to 250 MPa and a depth of layer greater than or equal to 10 μm. The glass article may have a type HgA1 hydrolytic resistance according to ISO 720.
As noted above, the presence of alkali oxides in the glass composition facilitates chemically strengthening the glass by ion exchange. Specifically, alkali ions, such as potassium ions, sodium ions and the like, are sufficiently mobile in the glass to facilitate ion exchange. In some embodiments, the glass composition is ion exchangeable to form a compressive stress layer having a depth of layer greater than or equal to 10 μm. In some embodiments, the depth of layer may be greater than or equal to about 25 μm or even greater than or equal to about 50 μm. In some other embodiments, the depth of the layer may be greater than or equal to 75 μm or even greater than or equal to 100 μm. In still other embodiments, the depth of layer may be greater than or equal to 10 μm and less than or equal to about 100 μm. The associated surface compressive stress may be greater than or equal to about 250 MPa, greater than or equal to 300 MPa or even greater than or equal to about 350 MPa after the glass composition is treated in a salt bath of 100% molten KNO3 at a temperature of 350° C. to 500° C. for a time period of less than about 30 hours or even about less than 20 hours.
The glass articles formed from the glass compositions described herein may have a hydrolytic resistance of HGB2 or even HGB1 under ISO 719 and/or a hydrolytic resistance of HGA2 or even HGA1 under ISO 720 (as described further herein) in addition to having improved mechanical characteristics due to ion exchange strengthening. In some embodiments described herein the glass articles may have compressive stresses which extend from the surface into the glass article to a depth of layer greater than or equal to 10 μm, greater than or equal to 15 μm, greater than or equal to 20 μm, greater than or equal to 25 μm, greater than or equal to 30 μm or even greater than or equal to 35 μm. In some embodiments, the depth of layer may be greater than or equal to 40 μm or even greater than or equal to 50 μm. The surface compressive stress of the glass article may be greater than or equal to 150 MPa, greater than or equal to 200 MPa, greater than or equal to 250 MPa, greater than or equal to 350 MPa, or even greater than or equal to 400 MPa.
In one embodiment, the glass pharmaceutical container has a compressive stress greater than or equal to 150 MPa and a depth of layer greater than or equal to 10 μm, 15 μm, 20 μm, 25 μm, 30 μm, 35 μm, 40 μm, 45 μm or 50 μm. In a particular embodiment, the glass pharmaceutical container has a compressive stress greater than or equal to 150 MPa and a depth of layer greater than or equal to 10 μm. In a particular embodiment, the glass pharmaceutical container has a compressive stress greater than or equal to 150 MPa and a depth of layer greater than or equal to 25 μm. In a particular embodiment, the glass pharmaceutical container has a compressive stress greater than or equal to 150 MPa and a depth of layer greater than or equal to 30 μm.
In one embodiment, the glass pharmaceutical container has a compressive stress greater than or equal to 300 MPa and a depth of layer greater than or equal to 10 μm, 15 μm, 20 μm, 25 μm, 30 μm, 35 μm, 40 μm, 45 μm or 50 μm. In a particular embodiment, the glass pharmaceutical container has a compressive stress greater than or equal to 300 MPa and a depth of layer greater than or equal to 25 μm. In yet another embodiment, the glass pharmaceutical container has a compressive stress greater than or equal to 300 MPa and a depth of layer greater than or equal to 30 μm. In yet another embodiment, the glass pharmaceutical container has a compressive stress greater than or equal to 300 MPa and a depth of layer greater than or equal to 35 μm.
The glass compositions described herein facilitate achieving the aforementioned depths of layer and surface compressive stresses more rapidly and/or at lower temperatures than conventional glass compositions due to the enhanced alkali ion diffusivity of the glass compositions as described hereinabove. For example, the depths of layer (i.e., greater than or equal to 25 μm) and the compressive stresses (i.e., greater than or equal to 250 MPa) may be achieved by ion exchanging the glass article in a molten salt bath of 100% KNO3 (or a mixed salt bath of KNO3 and NaNO3) for a time period of less than or equal to 5 hours or even less than or equal to 4.5 hours. In some embodiments, these depths of layer and compressive stresses may be achieved by ion exchanging the glass article in a molten salt bath of 100% KNO3 (or a mixed salt bath of KNO3 and NaNO3) for a time period of less than or equal to 4 hours or even less than or equal to 3.5 hours. Moreover, these depths of layers and compressive stresses may be achieved by ion exchanging the glass articles in a molten salt bath of 100% KNO3 (or a mixed salt bath of KNO3 and NaNO3) at a temperature less than or equal to 500° C. or even less than or equal to 450° C. In some embodiments, these depths of layers and compressive stresses may be achieved by ion exchanging the glass articles in a molten salt bath of 100% KNO3 (or a mixed salt bath of KNO3 and NaNO3) at a temperature less than or equal to 400° C. or even less than or equal to 350° C.
These improved ion exchange characteristics can be achieved when the glass composition has a threshold diffusivity of greater than about 16 μm2/hr or even greater than or equal to 20 μm2/hr at 450° C. In some embodiments, the threshold diffusivity may be greater than or equal to about 25 μm2/hr or even 30 μm2/hr at 450° C. In some other embodiments, the threshold diffusivity may be greater than or equal to about 35 μm2/hr or even 40 μm2/hr at 450° C. In still other embodiments, the threshold diffusivity may be greater than or equal to about 45 μm2/hr or even 50 μm2/hr at 450° C.
The glass compositions described herein may generally have a strain point greater than or equal to about 525° C. and less than or equal to about 650° C. The glasses may also have an anneal point greater than or equal to about 560° C. and less than or equal to about 725° C. and a softening point greater than or equal to about 750° C. and less than or equal to about 960° C.
In the embodiments described herein the glass compositions have a CTE of less than about 70×10−7K−1 or even less than about 60×10−7K−1. These lower CTE values improve the survivability of the glass to thermal cycling or thermal stress conditions relative to glass compositions with higher CTEs.
Further, as noted hereinabove, the glass compositions are chemically durable and resistant to degradation as determined by the DIN 12116 standard, the ISO 695 standard, and the ISO 720 standard.
Specifically, the DIN 12116 standard is a measure of the resistance of the glass to decomposition when placed in an acidic solution. In brief, the DIN 12116 standard utilizes a polished glass sample of a known surface area which is weighed and then positioned in contact with a proportional amount of boiling 6M hydrochloric acid for 6 hours. The sample is then removed from the solution, dried and weighed again. The glass mass lost during exposure to the acidic solution is a measure of the acid durability of the sample with smaller numbers indicative of greater durability. The results of the test are reported in units of half-mass per surface area, specifically mg/dm2. The DIN 12116 standard is broken into individual classes. Class S1 indicates weight losses of up to 0.7 mg/dm2; Class S2 indicates weight losses from 0.7 mg/dm2 up to 1.5 mg/dm2; Class S3 indicates weight losses from 1.5 mg/dm2 up to 15 mg/dm2; and Class S4 indicates weight losses of more than 15 mg/dm2.
The ISO 695 standard is a measure of the resistance of the glass to decomposition when placed in a basic solution. In brief, the ISO 695 standard utilizes a polished glass sample which is weighed and then placed in a solution of boiling 1M NaOH+0.5M Na2CO3 for 3 hours. The sample is then removed from the solution, dried and weighed again. The glass mass lost during exposure to the basic solution is a measure of the base durability of the sample with smaller numbers indicative of greater durability. As with the DIN 12116 standard, the results of the ISO 695 standard are reported in units of mass per surface area, specifically mg/dm2. The ISO 695 standard is broken into individual classes. Class A1 indicates weight losses of up to 75 mg/dm2; Class A2 indicates weight losses from 75 mg/dm2 up to 175 mg/dm2; and Class A3 indicates weight losses of more than 175 mg/dm2.
The ISO 720 standard is a measure of the resistance of the glass to degradation in purified, CO2-free water. In brief, the ISO 720 standard protocol utilizes crushed glass grains which are placed in contact with the purified, CO2-free water under autoclave conditions (121° C., 2 atm) for 30 minutes. The solution is then titrated colorimetrically with dilute HCl to neutral pH. The amount of HCl required to titrate to a neutral solution is then converted to an equivalent of Na2O extracted from the glass and reported in μg Na2O per weight of glass with smaller values indicative of greater durability. The ISO 720 standard is broken into individual types. Type HGA1 is indicative of up to 62 μg extracted equivalent of Na2O per gram of glass tested; Type HGA2 is indicative of more than 62 μg and up to 527 μg extracted equivalent of Na2O per gram of glass tested; and Type HGA3 is indicative of more than 527 μg and up to 930 μg extracted equivalent of Na2O per gram of glass tested.
The ISO 719 standard is a measure of the resistance of the glass to degradation in purified, CO2-free water. In brief, the ISO 719 standard protocol utilizes crushed glass grains which are placed in contact with the purified, CO2-free water at a temperature of 98° C. at 1 atmosphere for 30 minutes. The solution is then titrated colorimetrically with dilute HCl to neutral pH. The amount of HCl required to titrate to a neutral solution is then converted to an equivalent of Na2O extracted from the glass and reported in μg Na2O per weight of glass with smaller values indicative of greater durability. The ISO 719 standard is broken into individual types. The ISO 719 standard is broken into individual types. Type HGB1 is indicative of up to 31 μg extracted equivalent of Na2O; Type HGB2 is indicative of more than 31 μg and up to 62 μg extracted equivalent of Na2O; Type HGB3 is indicative of more than 62 μg and up to 264 μg extracted equivalent of Na2O; Type HGB4 is indicative of more than 264 μg and up to 620 μg extracted equivalent of Na2O; and Type HGB5 is indicative of more than 620 μg and up to 1085 μg extracted equivalent of Na2O. The glass compositions described herein have an ISO 719 hydrolytic resistance of type HGB2 or better with some embodiments having a type HGB1 hydrolytic resistance.
The glass compositions described herein have an acid resistance of at least class S3 according to DIN 12116 both before and after ion exchange strengthening with some embodiments having an acid resistance of at least class S2 or even class S1 following ion exchange strengthening. In some other embodiments, the glass compositions may have an acid resistance of at least class S2 both before and after ion exchange strengthening with some embodiments having an acid resistance of class S1 following ion exchange strengthening. Further, the glass compositions described herein have a base resistance according to ISO 695 of at least class A2 before and after ion exchange strengthening with some embodiments having a class A1 base resistance at least after ion exchange strengthening. The glass compositions described herein also have an ISO 720 type HGA2 hydrolytic resistance both before and after ion exchange strengthening with some embodiments having a type HGA1 hydrolytic resistance after ion exchange strengthening and some other embodiments having a type HGA1 hydrolytic resistance both before and after ion exchange strengthening. The glass compositions described herein have an ISO 719 hydrolytic resistance of type HGB2 or better with some embodiments having a type HGB1 hydrolytic resistance. It should be understood that, when referring to the above referenced classifications according to DIN 12116, ISO 695, ISO 720 and ISO 719, a glass composition or glass article which has “at least” a specified classification means that the performance of the glass composition is as good as or better than the specified classification. For example, a glass article which has a DIN 12116 acid resistance of “at least class S2” may have a DIN 12116 classification of either S1 or S2.
The glass compositions described herein are formed by mixing a batch of glass raw materials (e.g., powders of SiO2, Al2O3, alkali oxides, alkaline earth oxides and the like) such that the batch of glass raw materials has the desired composition. Thereafter, the batch of glass raw materials is heated to form a molten glass composition which is subsequently cooled and solidified to form the glass composition. During solidification (i.e., when the glass composition is plastically deformable) the glass composition may be shaped using standard forming techniques to shape the glass composition into a desired final form. Alternatively, the glass article may be shaped into a stock form, such as a sheet, tube or the like, and subsequently reheated and formed into the desired final form.
In order to assess the long-term resistance of the glass container to delamination, an accelerated delamination test was utilized. The test is performed on glass containers after the containers have been ion-exchange strengthened. The test consisted of washing the glass container at room temperature for 1 minute and depyrogenating the container at about 320° C. for 1 hour. Thereafter a solution of 20 mM glycine with a pH of 10 in water is placed in the glass container to 80-90% fill, the glass container is closed, and rapidly heated to 100° C. and then heated from 100° C. to 121° C. at a ramp rate of 1 deg/min at a pressure of 2 atmospheres. The glass container and solution are held at this temperature for 60 minutes, cooled to room temperature at a rate of 0.5 deg./min and the heating cycle and hold are repeated. The glass container is then heated to 50° C. and held for two days for elevated temperature conditioning. After heating, the glass container is dropped from a distance of at least 18′′ onto a firm surface, such as a laminated tile floor, to dislodge any flakes or particles that are weakly adhered to the inner surface of the glass container.
Thereafter, the solution contained in the glass container is analyzed to determine the number of glass particles present per liter of solution. Specifically, the solution from the glass container is directly poured onto the center of a Millipore Isopore Membrane filter (Millipore #ATTP02500 held in an assembly with parts #AP1002500 and #M000025A0) attached to vacuum suction to draw the solution through the filter within 10-15 seconds. Particulate flakes are then counted by differential interference contrast microscopy (DIC) in the reflection mode as described in “Differential interference contrast (DIC) microscopy and modulation contrast microscopy” from Fundamentals of light microscopy and digital imaging. New York: Wiley-Liss, pp 153-168. The field of view is set to approximately 1.5 mm×1.5 mm and particles larger than 50 microns are counted manually. There are 9 such measurements made in the center of each filter membrane in a 3×3 pattern with no overlap between images. A minimum of 100 mL of solution is tested. As such, the solution from a plurality of small containers may be pooled to bring the total amount of solution to 100 mL. If the containers contain more than 10 mL of solution, the entire amount of solution from the container is examined for the presence of particles. For containers having a volume greater than 10 mL containers, the test is repeated for a trial of 10 containers formed from the same glass composition under the same processing conditions and the result of the particle count is averaged for the 10 containers to determine an average particle count. Alternatively, in the case of small containers, the test is repeated for a trial of 10 sets of 10 mL of solution, each of which is analyzed and the particle count averaged over the 10 sets to determine an average particle count. Averaging the particle count over multiple containers accounts for potential variations in the delamination behavior of individual containers.
It should be understood that the aforementioned test is used to identify particles which are shed from the interior wall(s) of the glass container due to delamination and not tramp particles present in the container from forming processes or particles which precipitate from the solution enclosed in the glass container as a result of reactions between the solution and the glass. Specifically, delamination particles may be differentiated from tramp glass particles due based on the aspect ratio of the particle (i.e., the ratio of the width of the particle to the thickness of the particle). Delamination produces particulate flakes or lamellae which are irregularly shaped and are typically >50 μm in diameter but often >200 μm. The thickness of the flakes is usually greater than about 100 nm and may be as large as about 1 μm. Thus, the minimum aspect ratio of the flakes is typically >50. The aspect ratio may be greater than 100 and sometimes greater than 1000. Particles resulting from delamination processes generally have an aspect ratio which is generally greater than about 50. In contrast, tramp glass particles will generally have a low aspect ratio which is less than about 3. Accordingly, particles resulting from delamination may be differentiated from tramp particles based on aspect ratio during observation with the microscope. Validation results can be accomplished by evaluating the heel region of the tested containers. Upon observation, evidence of skin corrosion/pitting/flake removal, as described in “Nondestructive Detection of Glass Vial Inner Surface Morphology with Differential Interference Contrast Microscopy” from Journal of Pharmaceutical Sciences 101(4), 2012, pages 1378-1384, is noted.
In the embodiments described herein, glass containers which average less than 3 glass particles with a minimum width of 50 μm and an aspect ratio of greater than 50 per trial following accelerated delamination testing are considered to have a delamination factor of 3. In the embodiments described herein, glass containers which average less than 2 glass particles with a minimum width of 50 μm and an aspect ratio of greater than 50 per trial following accelerated delamination testing are considered to have a delamination factor of 2. In the embodiments described herein, glass containers which average less than 1 glass particle with a minimum width of 50 μm and an aspect ratio of greater than 50 per trial following accelerated delamination testing are considered to have a delamination factor of 1. In the embodiments described herein, glass containers which have 0 glass particles with a minimum width of 50 μm and an aspect ratio of greater than 50 per trial following accelerated delamination testing are considered to have a delamination factor of 0. Accordingly, it should be understood that the lower the delamination factor, the better the resistance of the glass container to delamination. In the embodiments described herein, the glass containers have a delamination factor of 3 or lower (i.e., a delamination factor of 3, 2, 1 or 0).
Pharmaceutical Containers
In view of the chemical durability of the glass composition of the present invention, the glass compositions described herein are particularly well suited for use in designing pharmaceutical containers for storing, maintaining and/or delivering pharmaceutical compositions, such as liquids, solutions, powders, e.g., lyophilized powders, solids and the like. As used herein, the term “pharmaceutical container” refers to a composition designed to store, maintain and/or deliver a pharmaceutical composition. The pharmaceutical containers, as described herein, are formed, at least in part, of the delamination resistant glass compositions described above. Pharmaceutical containers of the present invention include, but are not limited to, VacutainersTM cartridges, syringes, ampoules, bottles, flasks, phials, tubes, beakers, vials, injection pens or the like. In a particular embodiment, the pharmaceutical container is a vial. In a particular embodiment, the pharmaceutical container is an ampoule. In a particular embodiment, the pharmaceutical container is an injection pen. In a particular embodiment, the pharmaceutical container is a tube. In a particular embodiment, the pharmaceutical container is a bottle. In a particular embodiment, the pharmaceutical container is a syringe.
Moreover, the ability to chemically strengthen the glass compositions through ion exchange can be utilized to improve the mechanical durability of pharmaceutical containers formed from the glass composition. Accordingly, it should be understood that, in at least one embodiment, the glass compositions are incorporated in a pharmaceutical container in order to improve the chemical durability and/or the mechanical durability of the pharmaceutical container.
Pharmaceutical Compositions
In various embodiments, the pharmaceutical container further includes a pharmaceutical composition comprising an active pharmaceutical ingredient (API). As used herein, the term “pharmaceutical composition” refers to a composition comprising an active pharmaceutical ingredient to be delivered to a subject, for example, for therapeutic, prophylactic, diagnostic, preventative or prognostic effect. In certain embodiments, the pharmaceutical composition comprises the active pharmaceutical ingredient and a pharmaceutically acceptable carrier. As used herein, “pharmaceutically acceptable carrier” includes any and all solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonic and absorption delaying agents, and the like that are physiologically compatible. Examples of pharmaceutically acceptable carriers include one or more of water, saline, phosphate buffered saline, dextrose, glycerol, ethanol and the like, as well as combinations thereof. In many cases, it may be preferable to include isotonic agents, for example, sugars, polyalcohols such as mannitol, sorbitol, or sodium chloride in the composition. Pharmaceutically acceptable carriers may further comprise minor amounts of auxiliary substances such as wetting or emulsifying agents, preservatives or buffers, which enhance the shelf life or effectiveness of the active pharmaceutical agent.
As used herein, the term “active pharmaceutical ingredient” or “API” refers a substance in a pharmaceutical composition that provides a desired effect, for example, a therapeutic, prophylactic, diagnostic, preventative or prognostic effect. In various embodiments, the active pharmaceutical ingredient can be any of a variety of substances known in the art, for example, a small molecule, a polypeptide mimetic, a biologic, an antisense RNA, a small interfering RNA (siRNA), etc.
For example, in a particular embodiment, the active pharmaceutical ingredient may be a small molecule. As used herein, the term “small molecule” includes any chemical or other moiety, other than polypeptides and nucleic acids, that can act to affect biological processes. Small molecules can include any number of therapeutic agents presently known and used, or that can be synthesized from a library of such molecules for the purpose of screening for biological function(s). Small molecules are distinguished from macromolecules by size. The small molecules of the present invention usually have a molecular weight less than about 5,000 daltons (Da), preferably less than about 2,500 Da, more preferably less than 1,000 Da, most preferably less than about 500 Da.
Small molecules include, without limitation, organic compounds, peptidomimetics and conjugates thereof. As used herein, the term “organic compound” refers to any carbon-based compound other than macromolecules such as nucleic acids and polypeptides. In addition to carbon, organic compounds may contain calcium, chlorine, fluorine, copper, hydrogen, iron, potassium, nitrogen, oxygen, sulfur and other elements. An organic compound may be in an aromatic or aliphatic form. Non-limiting examples of organic compounds include acetones, alcohols, anilines, carbohydrates, monosaccharides, oligosaccharides, polysaccharides, amino acids, nucleosides, nucleotides, lipids, retinoids, steroids, proteoglycans, ketones, aldehydes, saturated, unsaturated and polyunsaturated fats, oils and waxes, alkenes, esters, ethers, thiols, sulfides, cyclic compounds, heterocyclic compounds, imidizoles, and phenols. An organic compound as used herein also includes nitrated organic compounds and halogenated (e.g., chlorinated) organic compounds.
In another embodiment, the active pharmaceutical ingredient may be a polypeptide mimetic (“peptidomimetic”). As used herein, the term “polypeptide mimetic” is a molecule that mimics the biological activity of a polypeptide, but that is not peptidic in chemical nature. While, in certain embodiments, a peptidomimetic is a molecule that contains no peptide bonds (that is, amide bonds between amino acids), the term peptidomimetic may include molecules that are not completely peptidic in character, such as pseudo-peptides, semi-peptides, and peptoids.
In other embodiments, the active pharmaceutical ingredient may be a biologic. As used herein, the term “biologic” includes products created by biologic processes instead of by chemical synthesis. Non-limiting examples of a “biologic” include proteins, antibodies, antibody like molecules, vaccines, blood, blood components, and partially purified products from tissues.
The terms “peptide,” “polypeptide,” and “protein” are used interchangeably herein. In the present invention, these terms mean a linked sequence of amino acids, which may be natural, synthetic, or a modification or combination of natural and synthetic. The term includes antibodies, antibody mimetics, domain antibodies, lipocalins, and targeted proteases. The term also includes vaccines containing a peptide or peptide fragment intended to raise antibodies against the peptide or peptide fragment.
“Antibody” as used herein includes an antibody of classes IgG, IgM, IgA, IgD, or IgE, or fragments or derivatives thereof, including Fab, F(ab′)2, Fd, and single chain antibodies, diabodies, bispecific antibodies, and bifunctional antibodies. The antibody may be a monoclonal antibody, polyclonal antibody, affinity purified antibody, or mixtures thereof, which exhibits sufficient binding specificity to a desired epitope or a sequence derived therefrom. The antibody may also be a chimeric antibody. The antibody may be derivatized by the attachment of one or more chemical, peptide, or polypeptide moieties known in the art. The antibody may be conjugated with a chemical moiety. The antibody may be a human or humanized antibody.
Other antibody-like molecules are also within the scope of the present invention. Such antibody-like molecules include, e.g., receptor traps (such as entanercept), antibody mimetics (such as adnectins, fibronectin based “addressable” therapeutic binding molecules from, e.g., Compound Therapeutics, Inc.), domain antibodies (the smallest functional fragment of a naturally occurring single-domain antibody (such as, e.g., nanobodies; see, e.g., Cortez-Retamozo et al., Cancer Res. 2004 Apr. 15; 64 (8):2853-7)).
Suitable antibody mimetics generally can be used as surrogates for the antibodies and antibody fragments described herein. Such antibody mimetics may be associated with advantageous properties (e.g., they may be water soluble, resistant to proteolysis, and/or be nonimmunogenic). For example, peptides comprising a synthetic beta-loop structure that mimics the second complementarity-determining region (CDR) of monoclonal antibodies have been proposed and generated. See, e.g., Saragovi et al., Science. Aug. 16, 1991; 253 (5021):792-5. Peptide antibody mimetics also have been generated by use of peptide mapping to determine “active” antigen recognition residues, molecular modeling, and a molecular dynamics trajectory analysis, so as to design a peptide mimic containing antigen contact residues from multiple CDRs. See, e.g., Cassett et al., Biochem Biophys Res Commun. Jul. 18, 2003; 307 (1):198-205. Additional discussion of related principles, methods, etc., that may be applicable in the context of this invention are provided in, e.g., Fassina, Immunomethods. October 1994; 5 (2):121-9.
In various embodiments, the active pharmaceutical ingredient may have any of a variety of activities selected from the group consisting of anti-rheumatics, anti-neoplastic, vaccines, anti-diabetics, haematologicals, muscle relaxant, immunostimulants, anti-coagulants, bone calcium regulators, sera and gammaglobulins, anti-fibrinolytics, MS therapies, anti-anaemics, cytostatics, interferons, anti-metabolites, radiopharmaceuticals, anti-psychotics, anti-bacterials, immunosuppressants, cytotoxic antibiotics, cerebral & peripheral vasotherapeutics, nootropics, CNS drugs, dermatologicals, angiotensin antagonists, anti-spasmodics, anti-cholinergics, interferons, anti-psoriasis agents, anti-hyperlipidaemics, cardiac therapies, alkylating agents, bronchodilators, anti-coagulants, anti-inflammatories, growth hormones, and diagnostic imaging agents.
In various embodiments, the pharmaceutical composition may be selected from the group consisting of PEDIARIX® (Diphtheria and Tetanus Toxoids and Acellular Pertussis Adsorbed, Hepatitis B (Recombinant) and Inactivated Poliovirus Vaccine), HAVRIX® (Hepatitis A Vaccine), ENGERIX-B® (Hepatitis B Vaccine (Recombinant)), TWINRIX® (Hepatitis A & Hepatitis B (Recombinant) Vaccine), EPERZAN® (albiglutide), MAGE-A3 Antigen-Specific Cancer Immunotherapeutic (astuprotimut-R), GSK2402968 (drisapersen), and HZ/su (herpes zoster vaccine).
In a particular embodiment, the pharmaceutical composition comprises Diphtheria and Tetanus Toxoids and Acellular Pertussis Adsorbed, Hepatitis B (Recombinant) and Inactivated Poliovirus Vaccine (PEDIARIX®). In a particular embodiment, the active pharmaceutical ingredient comprises diphtheria toxoid, tetanus toxoid, inactivated pertussis toxin, hepatitis B virus surface antigen, inactivated Type I poliovirus (Mahoney), Type 2 poliovirus (MEF-1), Type 3 poliovirus (Saukett), filamentous hemagglutinin and pertactin (69 kD outer membrane protein), or analogs thereof. PEDIARIX® is a form of noninfectious, sterile vaccine comprising diphtheria toxoid, tetanus toxoid, inactivated pertussis toxin, hepatitis B virus surface antigen, inactivated Type I poliovirus (Mahoney), Type 2 poliovirus (MEF-1), Type 3 poliovirus (Saukett), filamentous hemagglutinin and pertactin (69 kD outer membrane protein). Protection against disease is due to the development of neutralizing antibodies to the diphtheria toxin, the tetanus toxin, and to the poliovirus. The role of the different B. pertussis components (inactivated pertussis toxin, filamentous hemagglutinin and pertactin (69 kD outer membrane protein)) in the development of immunity to pertussis is not well understood. Antibody concentrations >10 mIU/mL against HBsAg confer protection against hepatitis B virus infection. PEDIARIX® is presently indicated for active immunization against diphtheria, tetanus, pertussis, infection caused by all known subtypes of hepatitis B virus, and poliomyelitis.
The diphtheria toxin and tetanus toxin are produced by growing Corynebacterium diphtheria and Clostridium tetani in culture, respectively, followed by detoxification of the toxins with formaldehyde. The toxins are concentrated by ultrafiltration and purified by precipitation, dialysis and sterile filtration. The pertussis antigens are isolated from Bordatella pertussis cells grown in culture. The pertussis toxin and filamentous hemagglutinin are isolated from the medium and the pertactin is isolated from the cells. All antigens are purified by chromatography and precipitation. The pertussis toxin is inactivated using glutaraldehye and formaldehye and the filamentous hemagglutinin and pertactin are treated with formaldehyde. The hepatitis B surface antigen is derived from recombinant Saccharomyces cerevisiae cells which carry the surface antigen gene of the hepatitis B virus. The surface antigen is purified by precipitation, ion exchange chromatography and ultrafiltration. The three strains of poliovirus are grown individually in VERO cells followed by purification by ultrafiltration, diafiltration and chromatography and inactivated using formaldehyde. The individually purified viruses are combined into a trivalent vaccine concentrate.
PEDIARIX® is a noninfectious, sterile vaccine for intramuscular administration. Three doses of 0.5 ml each are administered at 2, 4 and 6 months of age. Each 0.5-ml dose contains 25 Lf of diphtheria toxoid, 10 Lf of tetanus toxoid, 25 mcg of inactivated pertussis toxin (PT), 25 mcg of filamentous hemagglutinin (FHA), 8 mcg of pertactin (69 kDa outer membrane protein), 10 mcg of HBsAg, 40 D-antigen Units (DU) of Type 1 poliovirus (Mahoney), 8 DU of Type 2 poliovirus (MEF-1), and 32 DU of Type 3 poliovirus (Saukett). Each 0.5-ml dose contains not more than 0.85 mg aluminum salts as adjuvant and 4.5 mg sodium chloride. Each dose also contains ≦100 mcg of residual formaldehyde, ≦100 mcg of polysorbate 80 (Tween 80), and ≦5% yeast protein. Neomycin sulfate and polymyxin B may be present in the final vaccine at ≦0.05 ng and ≦0.01 ng per dose, respectively. PEDIARIX® is a suspension for injection available in 0.5 ml single-dose disposable prefilled syringe.
In a particular embodiment, the pharmaceutical composition comprises Hepatitis A Vaccine. In a particular embodiment, the active pharmaceutical ingredient comprises inactivated hepatitis A virus, or an analog thereof. Hepatitis A Vaccine (HAVRIX®) is a form of sterile suspension of inactivated hepatitis A virus (strain HM175). The presence of antibodies to hepatitis A virus confers protection against hepatitis A disease. HAVRIX® is indicated for active immunization against disease caused by hepatitis A virus infection.
The hepatitis A virus is propagated in MRC-5 human diploid cells and inactivated with formalin. Viral antigen activity is referenced to a standard using an enzyme linked immunosorbent assay (ELISA) and is expressed in terms of ELISA Units.
HAVRIX® is a sterile vaccine for intramuscular administration. Adults receive a single 1 ml dose and a 1 ml booster dose between 6 to 12 months later. Children and adolescents receive a single 0.5 ml dose and a 0.5 ml booster dose between 6 to 12 months later. Each 1 ml dose contains 1440 ELISA Units of viral antigen adsorbed onto 0.5 mg of aluminum as aluminum hydroxide. Each 0.5 ml dose contains 720 ELISA Units of viral antigen adsorbed onto 0.25 mg of aluminum as aluminum hydroxide. The dose also contains amino acid supplement (0.3% w/v) in a phosphate-buffered saline solution and polysorbate 20 (0.05 mg/ml). Residual compounds from the manufacturing process include not more than 5 mcg/ml residual MRC-5 cellular sulfate, not more than 0.1 mg/ml formalin and not more than 40 ng/ml neomycin sulfate. HAVRIX® is currently supplied in either a 0.5 ml or 1 ml single-dose vial and in a 0.5 ml or 1 ml prefilled syringe. HAVRIX® is a homogeneous turbid white suspension for injection.
In a particular embodiment, the pharmaceutical composition comprises Hepatitis B Vaccine (Recombinant). In a particular embodiment, the active pharmaceutical ingredient comprises noninfectious hepatitis B virus surface antigen (HBsAg), or analogs thereof. Hepatitis B Vaccine (Recombinant) (ENGERIX-B®) is a form of sterile suspension of noninfectious hepatitis B virus surface antigen (HBsAg). Protection against hepatitis B virus infection is due to antibody concentrations >10 mIU/mL against HBsAg. Seroconversion is defined as antibody titers ≧1 mIU/ml. ENGERIX-B® is indicated for active immunization against infection caused by all known subtypes of hepatitis B virus.
The hepatitis B surface antigen is derived from recombinant Saccharomyces cerevisiae cells which carry the surface antigen gene of the hepatitis B virus. The surface antigen is purified by precipitation, ion exchange chromatography and ultrafiltration.
ENGERIX-B® is a sterile vaccine for intramuscular administration. Individuals 20 years of age and older receive a series of 3, 1 ml doses given on a 0, 1, and 6 month schedule. Individuals from birth to 19 years of age receive a series of 3, 0.5 ml doses given on a 0, 1, and 6 month schedule. Each 1 ml dose contains 20 mcg of HBsAg adsorbed on 0.5 mg aluminum as aluminum hydroxide. Each 0.5 ml dose contains 10 mcg of HBsAg adsorbed on 0.25 mg aluminum as aluminum hydroxide. Excipients include sodium chloride (9 mg/ml) and phosphate buffers (disodium phosphate dihydrate, 0.98 mg/ml; sodium dihydrogen phosphate dihydrate, 0.71 mg/ml). Each dose contains no more than 5% yeast protein. ENGERIX® is currently supplied in either a 0.5 ml or 1 ml single-dose vial and in a 0.5 ml or 1 ml prefilled syringe. ENGERIX® is a homogeneous turbid white suspension for injection.
In a particular embodiment, the pharmaceutical composition comprises Hepatitis A & Hepatitis B (Recombinant) Vaccine. In a particular embodiment, the active pharmaceutical ingredient comprises inactivated hepatitis A virus and noninfectious hepatitis B virus surface antigen (HBsAg), or analogs thereof. Hepatitis A & Hepatitis B (Recombinant) Vaccine (TWINRIX®) is a form of bivalent vaccine containing inactivated hepatitis A virus (strain HM175) and noninfectious hepatitis B virus surface antigen (HBsAg). The presence of antibodies to hepatitis A virus confers protection against hepatitis A disease. Protection against hepatitis B virus infection is due to antibody concentrations >10 mIU/mL against HBsAg. TWINRIX® is indicated for active immunization against disease caused by hepatitis A virus infection and infection by all known subtypes of hepatitis B virus.
The hepatitis A virus is propagated in MRC-5 human diploid cells and inactivated with formalin. Viral antigen activity is referenced to a standard using an enzyme linked immunosorbent assay (ELISA) and is expressed in terms of ELISA Units. The hepatitis B surface antigen is derived from recombinant Saccharomyces cerevisiae cells which carry the surface antigen gene of the hepatitis B virus. The surface antigen is purified by precipitation, ion exchange chromatography and ultrafiltration.
TWINRIX® is a sterile vaccine for intramuscular administration. Individuals receive a standard dosing regimen of 3, 1 ml doses at 0, 1, and 6 months. Under an accelerated dosing regimen, individuals receive 4, 1 ml doses on days 0, 7, and 21 to 30 followed by a booster dose at 12 months. Each 1 ml dose contains 720 ELISA Units of inactivated hepatitis A virus and 20 mcg of recombinant HBsAg protein. The 1 ml dose of vaccine also contains 0.45 mg of aluminum in the form of aluminum phosphate and aluminum hydroxide as adjuvants, amino acids, sodium chloride, phosphate buffer, polysorbate 20 and Water for Injection. Residual compounds from the manufacturing process include not more than 0.1 mg of formalin, not more than 2.5 mcg of MRC-5 cellular proteins, not more than 20 ng neomycin sulfate and not more than 5% yeast protein. TWINRIX® is currently supplied as a 1 ml single-dose vial and as a prefilled syringe and is formulated as a turbid white suspension for injection.
In particular embodiments, the pharmaceutical composition comprises albiglutide. In a particular embodiment, the active pharmaceutical ingredient comprises a glucagon-like peptide-1 (GLP-1) receptor agonist, or an analog thereof. Albiglutide (EPERZAN®) is a form of glucagon-like peptide-1 (GLP-1) receptor agonist. GLP-1 is secreted by the gastrointestinal tract during eating and increases insulin secretion, decreases glycemic excursion, delays gastric emptying and reduces food intake. Albiglutide is presently indicated for treatment of adults with type 2 diabetes.
Albiglutide is a GLP-1 receptor agonist comprising a dipeptidyl peptidase-4 resistant GLP-1 dimer fused in series to human albumin Native GLP-1 peptide is rapidly degraded while albiglutide has a longer duration of action (e.g. half-life of 4 to 7 days) due to its resistance to hydrolysis by dipeptidyl peptidase-4. Albiglutide is designed for weekly or biweekly subcutaneous dosing. Albiglutide may be provided at a concentration of 50 mg/mL following resuspension of a lyophilized form comprising 2.8% mannitol, 4.2% trehalose dihydrate, 0.01% polysorbate 80, and 10 to 20 mM phosphate buffer at pH 7.2. The formulation is diluted with water for injection as necessary for dosing.
In a particular embodiment, the pharmaceutical composition comprises astuprotimut-R. In a particular embodiment, the active pharmaceutical ingredient comprises a fusion protein consisting of the MAGE-A3 (melanoma associated antigen 3), or an analog thereof. Astuprotimut-R (MAGE-A3 Antigen-Specific Cancer Immunotherapeutic) is a form of fusion protein consisting of the MAGE-A3 (melanoma associated antigen 3) epitope. MAGE-A3 may also be referred to as MAGE-3. A MAGE-A3 fusion protein may induce a therapeutic effect by stimulating a cytotoxic T lymphocyte (CTL) response against tumor cells that express the MAGE-A3 antigen, resulting in tumor cell death. Astuprotimut-R is presently indicated for the treatment of cancer (e.g., melanoma, non-small cell lung cancer).
Astuprotimut-R is a fusion protein consisting of the MAGE-A3 (melanoma associated antigen 3) epitope fused to part of the Haemophilus influenza protein D antigen sequence. MAGE-A3 is a tumor specific antigen expressed in a variety of cancers (e.g., melanoma, non-small cell lung cancer, head and neck cancer and bladder cancer). Astuprotimut-R is administered along with an adjuvant system comprising specific combinations of immunostimulating compounds selected to increase the anti-tumor response.
In a particular embodiment, the pharmaceutical composition comprises drisapersen. In a particular embodiment, the active pharmaceutical ingredient is a dystrophin antisense oligonucleotide, or an analog thereof. Drisapersen (GSK2402968) is a form of dystrophin antisense oligonucleotide. Antisense oligonucleotides may affect the post-transcriptional splicing process of the premRNA to restore the reading frame of the mRNA, resulting in a shortened dystrophin protein. Drisapersen is currently indicated for the treatment of boys with Duchenne Muscular Dystrophy with a dystrophin gene mutation amenable to an exon 51 skip.
Drisapersen is a dystrophin antisense oligonucleotide (5′-UCAAGGAAGAUGGCAUUUCA-3′) (SEQ ID NO:1) with full length 2′-O-methyl-substituted ribose moieties and phosphorothioate internucleotide linkages. Drisapersen is designed to cause skipping of exon 51 of the dystrophin pre-mRNA and to restore the reading frame. Binding of the antisense oligonucleotide to either one or both splice sites or exon-internal sequences induces skipping of the exon by blocking recognition of the splice sites by the spliceosome complex.
Drisapersen may be provided in 0.5 ml glass vial in sodium phosphate buffered saline at a concentration of 100 mg/ml. It may be formulated for abdominal subcutaneous administration at a dose of 0.5 to 10 mg/kg of body weight.
In a particular embodiment, the pharmaceutical composition comprises a herpes zoster vaccine. In a particular embodiment, the active pharmaceutical composition comprises a recombinant varicella zoster virus glycoprotein E, or an analog thereof. Herpes zoster vaccine (HZ/su) is a form of recombinant adjuvated vaccine comprising the varicella zoster virus glycoprotein E. The varicella zoster virus glycoprotein E subunit is the most abundant glycoprotein in the varicella zoster virus and induces a potent CD4+ T-cell response when administered with an adjuvant. Herpes zoster vaccine is presently indicated for prevention of herpes zoster infection and its complications.
Herpes virus vaccine is formulated for intramuscular injection and comprises 50 μg of varicella zoster virus glycoprotein E in 0.2 ml mixed with 0.5 ml of AS01B adjuvant. AS01B adjuvant is a liposome based adjuvant system containing 50 μg 3-O-desacyl-4′-monophosphoryl lipid A (MPL) and 50 μg QS21 (a triterpene glycoside).
Degradation and Stability of Pharmaceutical Compositions
According to the present invention, delamination resistant pharmaceutical containers comprising a glass composition provide for improved resistance to degradation of, improved stability of, improved resistance to inactivation of, and improved maintenance of levels of a pharmaceutical composition having at least one active pharmaceutical ingredient, for example, PEDIARIX® (Diphtheria and Tetanus Toxoids and Acellular Pertussis Adsorbed, Hepatitis B (Recombinant) and Inactivated Poliovirus Vaccine), HAVRIX® (Hepatitis A Vaccine), ENGERIX-B® (Hepatitis B Vaccine (Recombinant)), TWINRIX® (Hepatitis A & Hepatitis B (Recombinant) Vaccine), EPERZAN® (albiglutide), MAGE-A3 Antigen-Specific Cancer Immunotherapeutic (astuprotimut-R), GSK2402968 (drisapersen), or HZ/su (herpes zoster vaccine).
In one embodiment of the present invention, the delamination resistant pharmaceutical containers provide improved stability to pharmaceutical compositions contained therein, for example, PEDIARIX® (Diphtheria and Tetanus Toxoids and Acellular Pertussis Adsorbed, Hepatitis B (Recombinant) and Inactivated Poliovirus Vaccine), HAVRIX® (Hepatitis A Vaccine), ENGERIX-B® (Hepatitis B Vaccine (Recombinant)), TWINRIX® (Hepatitis A & Hepatitis B (Recombinant) Vaccine), EPERZAN® (albiglutide), MAGE-A3 Antigen-Specific Cancer Immunotherapeutic (astuprotimut-R), GSK2402968 (drisapersen), or HZ/su (herpes zoster vaccine). As used herein, the term “stability” refers to the ability of an active pharmaceutical ingredient to essentially retain its physical, chemical and conformational identity and integrity upon storage in the pharmaceutical containers of the invention. Stability is associated with the ability of an active pharmaceutical ingredient to retain its potency and efficacy over a period of time. Instability of an active pharmaceutical ingredient may be associated with, for example, chemical or physical degradation, fragmentation, conformational change, increased toxicity, aggregation (e.g., to form higher order polymers), deglycosylation, modification of glycosylation, oxidation, hydrolysis, or any other structural, chemical or physical modification. Such physical, chemical and/or conformational changes often result in reduced activity or inactivation of the active pharmaceutical ingredient, for example, such that at least one biological activity of the active pharmaceutical ingredient is reduced or eliminated. Alternatively or in addition, such physical, chemical and/or conformational changes often result in the formation of structures toxic to the subject to whom the pharmaceutical composition is administered.
The pharmaceutical containers of the present invention maintain stability of the pharmaceutical compositions, in part, by minimizing or eliminating delamination of the glass composition which forms, at least in part, the pharmaceutical container. In addition, the pharmaceutical containers of the present invention maintain stability of the pharmaceutical compositions, in part, by reducing or preventing the interaction of the active pharmaceutical ingredient with the pharmaceutical container and/or delaminated particles resulting therefrom. By minimizing or eliminating delamination and, further, by reducing or preventing interaction, the pharmaceutical containers thereby reduce or prevent the destabilization of the active pharmaceutical ingredient as found in, for example, PEDIARIX® (Diphtheria and Tetanus Toxoids and Acellular Pertussis Adsorbed, Hepatitis B (Recombinant) and Inactivated Poliovirus Vaccine), HAVRIX® (Hepatitis A Vaccine), ENGERIX-B® (Hepatitis B Vaccine (Recombinant)), TWINRIX® (Hepatitis A & Hepatitis B (Recombinant) Vaccine), EPERZAN® (albiglutide), MAGE-A3 Antigen-Specific Cancer Immunotherapeutic (astuprotimut-R), GSK2402968 (drisapersen), or HZ/su (herpes zoster vaccine).
The pharmaceutical containers of the present invention provide the additional advantage of preventing loss of active pharmaceutical ingredients. For example, by reducing or preventing the interaction of and, thus, the adherence of, the active pharmaceutical ingredient with the pharmaceutical container and/or delaminated particles resulting therefrom, the level of active pharmaceutical ingredient available for administration to a subject is maintained, as found in, for example, PEDIARIX® (Diphtheria and Tetanus Toxoids and Acellular Pertussis Adsorbed, Hepatitis B (Recombinant) and Inactivated Poliovirus Vaccine), HAVRIX® (Hepatitis A Vaccine), ENGERIX-B® (Hepatitis B Vaccine (Recombinant)), TWINRIX® (Hepatitis A & Hepatitis B (Recombinant) Vaccine), EPERZAN® (albiglutide), MAGE-A3 Antigen-Specific Cancer Immunotherapeutic (astuprotimut-R), GSK2402968 (drisapersen), or HZ/su (herpes zoster vaccine).
In one embodiment of the present invention, the pharmaceutical composition has a high pH. According to the present invention, it has been discovered that high pHs serve to increase delamination of glass compositions. Accordingly, the pharmaceutical containers of the present invention are particularly suitable for storing and maintaining pharmaceutical compositions having a high pH, for example, pharmaceutical compositions having a pH between about 7 and about 11, between about 7 and about 10, between about 7 and about 9, or between about 7 and about 8.
In additional embodiments, the pharmaceutical containers of the present invention are particularly suitable for storing and maintaining pharmaceutical compositions having phosphate or citrate based buffers. According to the present invention, it has been discovered that phosphate or citrate based buffers serve to increase delamination of glass compositions. According in particular embodiments, the pharmaceutical composition includes a buffer comprising a salt of citrate, e.g., sodium citrate, or SSC. In other embodiments, the pharmaceutical composition includes a buffer comprising a salt of phosphate, e.g., mono or disodium phosphate.
In additional embodiments, the pharmaceutical containers of the present invention are particularly suitable for storing and maintaining active pharmaceutical ingredient that needs to be subsequently formulated. In other embodiments, the pharmaceutical containers of the present invention are particularly suitable for storing and maintaining a lyophilized pharmaceutical composition or active pharmaceutical ingredient that requires reconstitution, for example, by addition of saline.
Assaying for Delamination of Pharmaceutical Containers
As noted above, delamination may result in the release of silica-rich glass flakes into a solution contained within the glass container after extended exposure to the solution. Accordingly, the resistance to delamination may be characterized by the number of glass particulates present in a solution contained within the glass container after exposure to the solution under specific conditions. In order to assess the long-term resistance of the glass container to delamination, an accelerated delamination test was utilized. The test consisted of washing the glass container at room temperature for 1 minute and depyrogenating the container at about 320° C. for 1 hour. Thereafter a solution of 20 mM glycine with a pH of 10 in water is placed in the glass container to 80-90% fill, the glass container is closed, and rapidly heated to 100° C. and then heated from 100° C. to 121° C. at a ramp rate of 1 deg/min at a pressure of 2 atmospheres. The glass container and solution are held at this temperature for 60 minutes, cooled to room temperature at a rate of 0.5 deg./min and the heating cycle and hold are repeated. The glass container is then heated to 50° C. and held for two days for elevated temperature conditioning. After heating, the glass container is dropped from a distance of at least 18′′ onto a firm surface, such as a laminated tile floor, to dislodge any flakes or particles that are weakly adhered to the inner surface of the glass container.
Thereafter, the solution contained in the glass container is analyzed to determine the number of glass particles present per liter of solution. Specifically, the solution from the glass container is directly poured onto the center of a Millipore Isopore Membrane filter (Millipore #ATTP02500 held in an assembly with parts #AP1002500 and #M000025A0) attached to vacuum suction to draw the solution through the filter within 10-15 seconds. Particulate flakes are then counted by differential interference contrast microscopy (DIC) in the reflection mode as described in “Differential interference contrast (DIC) microscopy and modulation contrast microscopy” from Fundamentals of light microscopy and digital imaging. New York: Wiley-Liss, pp 153-168. The field of view is set to approximately 1.5 mm×1.5 mm and particles larger than 50 microns are counted manually. There are 9 such measurements made in the center of each filter membrane in a 3×3 pattern with no overlap between images. A minimum of 100 mL of solution is tested. As such, the solution from a plurality of small containers may be pooled to bring the total amount of solution to 100 mL. If the containers contain more than 10 mL of solution, the entire amount of solution from the container is examined for the presence of particles. For containers having a volume greater than 10 mL, the test is repeated for a trial of 10 containers formed from the same glass composition under the same processing conditions and the result of the particle count is averaged for the 10 containers to determine an average particle count. Alternatively, in the case of small containers, the test is repeated for a trial of 10 sets of 10 mL of solution, each of which is analyzed and the particle count averaged over the 10 sets to determine an average particle count. Averaging the particle count over multiple containers accounts for potential variations in the delamination behavior of individual containers. Table 7 summarizes some non-limiting examples of sample volumes and numbers of containers for testing is shown below:







TABLE 7







Table of Exemplary Test Specimens












Nominal




Total


Vial
Vial Max
Minimum
Number of

solution


Capacity
Volume
Solution per
Vials in a
Number of
Tested


(mL)
(mL)
Vial (mL)
Trial
Trials
(mL)















2
4
3.2
4
10
128


3.5
7
5.6
2
10
112


4
6
4.8
3
10
144


5
10
8
2
10
160


6
10
8
2
10
160


8
11.5
9.2
2
10
184


10
13.5
10.8
1
10
108


20
26
20.8
1
10
208


30
37.5
30
1
10
300


50
63
50.4
1
10
504









It should be understood that the aforementioned test is used to identify particles which are shed from the interior wall(s) of the glass container due to delamination and not tramp particles present in the container from forming processes or particles which precipitate from the solution enclosed in the glass container as a result of reactions between the solution and the glass. Specifically, delamination particles may be differentiated from tramp glass particles based on the aspect ratio of the particle (i.e., the ratio of the width of the particle to the thickness of the particle). Delamination produces particulate flakes or lamellae which are irregularly shaped and are typically >50 μm in diameter but often >200 μm. The thickness of the flakes is usually greater than about 100 nm and may be as large as about 1 μm. Thus, the minimum aspect ratio of the flakes is typically >50. The aspect ratio may be greater than 100 and sometimes greater than 1000. Particles resulting from delamination processes generally have an aspect ratio which is generally greater than about 50. In contrast, tramp glass particles will generally have a low aspect ratio which is less than about 3. Accordingly, particles resulting from delamination may be differentiated from tramp particles based on aspect ratio during observation with the microscope. Validation results can be accomplished by evaluating the heel region of the tested containers. Upon observation, evidence of skin corrosion/pitting/flake removal, as described in “Nondestructive Detection of Glass Vial Inner Surface Morphology with Differential Interference Contrast Microscopy” from Journal of Pharmaceutical Sciences 101(4), 2012, pages 1378-1384, is noted.
In the embodiments described herein, glass containers which average less than 3 glass particles with a minimum width of 50 μm and an aspect ratio of greater than 50 per trial following accelerated delamination testing are considered “delamination resistant.” In the embodiments described herein, glass containers which average less than 2 glass particles with a minimum width of 50 μm and an aspect ratio of greater than 50 per trial following accelerated delamination testing are considered “delamination-stable.” In the embodiments described herein, glass containers which average less than 1 glass particle with a minimum width of 50 μm and an aspect ratio of greater than 50 per trial following accelerated delamination testing are considered “delamination-proof.” In the embodiments described herein, glass containers which have 0 glass particles with a minimum width of 50 μm and an aspect ratio of greater than 50 per trial following accelerated delamination testing are considered “delamination-free”.
Assessing Stability of Pharmaceutical Compositions
As set forth above, any of a variety of active pharmaceutical ingredients can be incorporated within the claimed pharmaceutical container including, for example, a small molecule, a polypeptide mimetic, a biologic, an antisense RNA, a small interfering RNA (siRNA), etc. These active ingredients degrade in varying manners and, thus, assessing the stability thereof in the pharmaceutical containers of the present invention requires different techniques.
Depending on the nature of the active pharmaceutical ingredient, the stability, maintenance and/or continued efficacy of the pharmaceutical compositions contained within the delamination resistant pharmaceutical containers of the present invention can be evaluated as follows.
Biologics API are often susceptible to degradation and/or inactivation arising from various factors, including pH, temperature, temperature cycling, light, humidity, etc. Biologics API are further susceptible to degradation, inactivation or loss, arising from interaction of the pharmaceutical composition with the pharmaceutical container, or delaminants leeching therefrom. For example, biologics may undergo physical degradation which may render the resulting pharmaceutical composition inactive, toxic or insufficient to achieve the desired effect. Alternatively, or in addition, biologics may undergo structural or conformational changes that can alter the activity of the API, with or without degradation. For example, proteins may undergo unfolding which can result in effective loss and inactivity of the API. Alternatively, or in addition, biologics may adhere to the surface of the container, thereby rendering the API administered to the subject insufficient to achieve the desired effect, e.g., therapeutic effect.
(i) General Methods for Investigation of Biologic Compound Degradation
Depending on the size and complexity of the biologic, methods for analysis of degradation of non-biologic, small molecule API may be applied to biologics. For example, peptides and nucleic acids can be analyzed using any of a number of chromatography and spectrometry techniques applicable to small molecules to determine the size of the molecules, either with or without protease or nuclease digestion. However, as proper secondary and tertiary structures are required for the activity of biologics, particularly protein biologics, confirmation of molecular weight is insufficient to confirm activity of biologics. Protein biologics containing complex post-translational modifications, e.g., glycosylation, are less amenable to analysis using chromatography and spectrometry. Moreover, complex biologics, e.g., vaccines which can include complex peptide mixtures, attenuated or killed viruses, or killed cells, are not amenable to analysis by most chromatography or spectrometry methods.
(ii) In Vitro Functional Assays for Investigation of Compound Stability
One or more in vitro assays, optionally in combination with one or more in vivo assays, can be used to assess the stability and activity of the API. Functional assays to determine API stability can be selected based on the structural class of the API and the function of the API. Exemplary assays are provided below to confirm the activity of the API after stability and/or stress testing. It is understood that assays should be performed with the appropriate controls (e.g., vehicle controls, control API not subject to stress or stability testing) with a sufficient number of dilutions and replicate samples to provide data with sufficient statistical significance to detect changes in activity of 10% or less, preferably 5% or less, 4% or less, more preferably 3% or less, 2% or less, or 1% or less, as desired. Such considerations in the art are well understood.
For example, antibody based therapeutics, regardless of the disease or condition to be treated, can be assayed for stability and activity using assays that require specific binding of the antibody to its cognate antigen, e.g., ELISA. The antigen used in the ELISA should have the appropriate conformational structure as would be found in vivo. Antibody based API are used, for example, for the treatment of cancer and inflammatory diseases including autoimmune diseases.
ELISA assays to assay the concentration of a protein biologic API are commercially available from a number of sources, e.g., R&D Systems, BD Biosciences, AbCam, Pierce, Invitrogen.
API are frequently targeted to receptors, particularly cell surface receptors. Receptor binding assays can be used to assess the activity of such agents. API that bind cell surface receptors can be agonists, antagonists or allosteric modulators. API that bind cell surface receptors need not bind the same location as the native ligand to modulate, for example, inhibit or enhance, signaling through the receptor. Depending on the activity of the API, an appropriate assay can be selected, e.g., assay for stimulation of receptor signaling when the API is a receptor agonist; and inhibition assay in which binding of an agonist, e.g., inhibition of activation by a receptor agonist by the API. Such assays can be used regardless of the disease(s) or condition(s) to be treated with the API. Modulation of cellular activity, e.g., cell proliferation, apoptosis, cell migration, modulation of expression of genes or proteins, differentiation, tube formation, etc. is assayed using routine methods. In other assay methods, a reporter construct is used to indicate activation of the receptor. Such methods are routine in the art. APIs that bind to cell surface receptors are used, for example, as anti-cancer agents, anti-diabetic agents, anti-inflammatory agents for the treatment of inflammatory mediated diseases including autoimmune disorders, anti-angiogenic agents, anti-cholinergic agents, bone calcium regulators, muscle and vascular tension regulators, and psychoactive agents.
Modulators of cell proliferation can be assayed for activity using a cell proliferation assays. For example, cell proliferation is induced using anti-anemic agents or stimulators of hematopoietic cell growth. Anti-proliferative agents, e.g., cytotoxic agents, anti-neoplastic agents, chemotherapeutic agents, cytostatic agents, antibiotic agents, are used to inhibit growth of various cell types. Some anti-inflammatory agents also act by inhibiting proliferation of immune cells, e.g., blast cells. In proliferation assays, replicate wells containing the same number of cells are cultured in the presence of the API. The effect of the API is assessed using, for example, microscopy or fluorescence activated cell sorting (FACS) to determine if the number of cells in the sample increased or decreased in response to the presence of the API. It is understood that the cell type selected for the proliferation assay is dependent on the specific API to be tested.
Modulators of angiogenesis can be assayed using cell migration and/or tube formation assays. For cell migration assays, human vascular endothelial cells (HUVECs) are cultured in the presence of the API in transwell devices. Migration of cells through the device at the desired time intervals is assessed. Alternatively, 3-dimensional HUVECs cultures in MATRIGEL can be assessed for tube formation. Anti-angiogenic agents are used, for example, for the treatment of cancer, macular degeneration, and diabetic retinopathy.
Anti-inflammatory API can be assayed for their effects on immune cell stimulation as determined, for example, by modulation of one or more of cytokine expression and secretion, antigen presentation, migration in response to cytokine or chemokine stimulation, and immune cell proliferation. In such assays, immune cells are cultured in the presence of the API and changes in immune cell activity are determined using routine methods in the art, e.g., ELISA and cell imaging and counting.
Anti-diabetic API can be assayed for their effects on insulin signaling, including insulin signaling in response to modulated glucose levels, and insulin secretion. Insulin signaling can be assessed by assessing kinase activation in response to exposure to insulin and/or modulation of glucose levels. Insulin secretion can be assessed by ELISA assay.
Modulators of blood clotting, i.e., fibrinolytics, anti-fibrinolytics, and anti-coagulants, can be assayed for their effects using an INR assay on serum by measuring prothrombin time to determine a prothrombin ratio. Time to formation of a clot is assayed in the presence or absence of the API.
Modulators of muscle or vascular tone can be assayed for their effects using vascular or muscle explants. The tissue can be placed in a caliper for detection of changes in length and/or tension. Whole coronary explants can be used to assess the activity of API on heart. The tissue is contacted with the API, and optionally agents to alter vascular tone (e.g., K+, Ca++). The effects of the API on length and/or tension of the vasculature or muscle is assessed.
Psychoactive agents can act by modulation of neurotransmitter release and/or recycling. Neuronal cells can be incubated in the presence of an API and stimulated to release neurotransmitters. Neurotransmitter levels can be assessed in the culture medium collected at defined time points to detect alterations in the level of neurotransmitter present in the media. Neurotransmitters can be detected, for example, using ELISA, LC/MS/MS, or by preloading the vesicles with radioactive neurotransmitters to facilitate detection.
(iii) In Vivo Assays for Investigation of Compound Stability
In addition to in vitro testing for compound stability, API can also be tested in vivo to confirm the stability of the API after storage and/or stress testing. For example, some API are not amenable to testing using in vitro assays due to the complexity of the disease state or the complexity of the response required. For example, psychoactive agents, e.g., antipsychotic agents, anti-depressant agents, nootropic agents, immunosuppressant agents, vasotherapeutic agents, muscular dystrophy agents, central nervous system modulating agents, antispasmodic agents, bone calcium regenerating agents, anti-rheumatic agents, anti-hyperlipidemic agents, hematopoietic proliferation agents, growth factors, vaccine agents, and imaging agents, may not be amenable to full functional characterization using in vitro models. Moreover, for some agents, factors that may not alter in vitro activity may alter activity in vivo, e.g., antibody variable domains may be sufficient to block signaling through a receptor, but the Fc domains may be required for efficacy in the treatment of disease. Further, changes in stability may result in changes in pharmacokinetic properties of an API (e.g., half-life, serum protein binding, tissue distribution, CNS permeability). Finally, changes in stability may result in the generation of toxic degradation or reaction products that would not be detected in vivo. Therefore, confirmation of pharmacokinetic and pharmacodynamic properties and toxicity in vivo is useful in conjunction with stability and stress testing.
(iv) Pharmacokinetic Assays
Pharmacokinetics includes the study of the mechanisms of absorption and distribution of an administered drug, the rate at which a drug action begins and the duration of the effect, the chemical changes of the substance in the body (e.g. by metabolic enzymes such as CYP or UGT enzymes) and the effects and routes of excretion of the metabolites of the drug. Pharmacokinetics is divided into several areas including the extent and rate of absorption, distribution, metabolism and excretion. This is commonly referred to as the ADME scheme:

    
    
        Absorption—the process of a substance entering the blood circulation.
        Distribution—the dispersion or dissemination of substances throughout the fluids and tissues of the body.
        Metabolism (or Biotransformation)—the irreversible transformation of parent compounds into daughter metabolites.
        Excretion—the removal of the substances from the body. In rare cases, some drugs irreversibly accumulate in body tissue.
        Elimination is the result of metabolism and excretion.
    
    


Pharmacokinetics describes how the body affects a specific drug after administration. Pharmacokinetic properties of drugs may be affected by elements such as the site of administration and the dose of administered drug, which may affect the absorption rate. Such factors cannot be fully assessed using in vitro models.
The specific pharmacokinetic properties to be assessed for a specific API in stability testing will depend, for example, on the specific API to be tested. In vitro pharmacokinetic assays can include assays of drug metabolism by isolated enzymes or by cells in culture. However, pharmacokinetic analysis typically requires analysis in vivo.
As pharmacokinetics are not concerned with the activity of the drug, but instead with the absorption, distribution, metabolism, and excretion of the drug, assays can be performed in normal subjects, rather than subjects suffering from a disease or condition for which the API is typically administered, by administration of a single dose of the API to the subject. However, if the subject to be treated with the API has a condition that would alter the metabolism or excretion of the API, e.g., liver disease, kidney disease, testing of the API in an appropriate disease model may be useful. Depending on the half-life of the compound, samples (e.g., serum, urine, stool) are collected at predetermined time points for at least two, preferably three half-lives of the API, and analyzed for the presence of the API and metabolic products of the API. At the end of the study, organs are harvested and analyzed for the presence of the API and metabolic products of the API. The pharmacokinetic properties of the API subjected to stability and/or stress testing are compared to API not subjected to stability or stress testing and other appropriate controls (e.g., vehicle control). Changes in pharmacokinetic properties as a result of stability and/or stress testing are determined.
(v) Pharmacodynamic Assays
Pharmacodynamics includes the study of the biochemical and physiological effects of drugs on the body or on microorganisms or parasites within or on the body and the mechanisms of drug action and the relationship between drug concentration and effect. Due to the complex nature of many disease states and the actions of many API, the API should be tested in vivo to confirm the desired activity of the agent. Mouse models for a large variety of disease states are known and commercially available (see, e.g., the Jackson Laboratory). A number of induced models of disease are also known. Agents can be tested on the appropriate animal model to demonstrate stability and efficacy of the API on modulating the disease state.
(vi) Specific Immune Response Assay
Vaccines produce complex immune responses that are best assessed in vivo. The specific potency assay for a vaccine depends, at least in part, on the specific vaccine type. The most accurate predictions are based on mathematical modeling of biologically relevant stability-indicating parameters. For complex vaccines, e.g., whole cell vaccines, whole virus vaccines, complex mixtures of antigens, characterization of each component biochemically may be difficult, if not impossible. For example, when using a live, attenuated virus vaccine, the number of plaque forming units (e.g., mumps, measles, rubella, smallpox) or colony forming units (e.g., S. typhi, TY21a) are determined to confirm potency after storage. Chemical and physical characterization (e.g., polysaccharide and polysaccharide-protein conjugate vaccines) is performed to confirm the stability and activity of the vaccine. Serological response in animals to inactivated toxins and/or animal protection against challenge (e.g., rabies, anthrax, diphtheria, tetanus) is performed to confirm activity of vaccines of any type, particularly when the activity of the antigen has been inactivated. In vivo testing of vaccines subjected to stability and/or stress testing is performed by administering the vaccine to a subject using the appropriate immunization protocol for the vaccine, and determining the immune response by detection of specific immune cells that respond to stimulation with the antigen or pathogen, detection of antibodies against the antigen or pathogen, or protection in an immune challenge. Such methods are well known in the art. Vaccines include, but are not limited to, meningococcal B vaccine, hepatitis A and B vaccines, human papillomavirus vaccine, influenza vaccine, herpes zoster vaccine, and pneumococcal vaccine.
(vii) Toxicity Assays
Degradation of API can result in in the formation of toxic agents. Toxicity assays include the administration of doses, typically far higher than would be used for therapeutic applications, to detect the presence of toxic products in the API. Toxicity assays can be performed in vitro and in vivo and are frequently single, high dose experiments. After administration of the compound, in addition to viability, organs are harvested and analyzed for any indication of toxicity, especially organs involved with clearance of API, e.g., liver, kidneys, and those for which damage could be catastrophic, e.g., heart, brain. The toxicologic properties of the API subjected to stability and/or stress testing are compared to API not subjected to stability or stress testing and other appropriate controls (e.g., vehicle control). Changes in toxicologic properties as a result of stability and/or stress testing are determined.
In accordance with present invention, the degradation, alteration or depletion of API contained within a delamination resistant pharmaceutical container of the present invention can be assessed by a variety of physical techniques. Indeed, in various aspects of the invention, the stability and degradation caused by the interaction of API with the container or delaminants thereof, or changes in concentration or amount of the API in a container can be assessed using techniques as follows. Such methods include, e.g., X-Ray Diffraction (XRPD), Thermal Analysis (such as Differential Scanning calorimetry (DSC), Thermogravimetry (TG) and Hot-Stage Microscopy (HSM), chromatography methods (such as High-Performance Liquid Chromatography (HPLC), Column Chromatography (CC), Gas Chromatography (GC), Thin-Layer Chromatography (TLC), and Super Critical Phase Chromatograph (SFC)), Mass Spectroscopy (MS), Capillary Electrophoresis (CE), Atomic Spectroscopy (AS), vibrational spectroscopy (such as Infrared Spectroscopy (IR)), Luminescence Spectroscopy (LS), and Nuclear Magnetic Resonance Spectroscopy (NMR).
In the case of pharmaceutical formulations where the API is not in solution or needs to be reconstituted into a different medium, XRPD may be a method for analyzing degradation. In ideal cases, every possible crystalline orientation is represented equally in a non-liquid sample.
Powder diffraction data is usually presented as a diffractogram in which the diffracted intensity I is shown as function either of the scattering angle 2θ or as a function of the scattering vector q. The latter variable has the advantage that the diffractogram no longer depends on the value of the wavelength λ. Relative to other methods of analysis, powder diffraction allows for rapid, non-destructive analysis of multi-component mixtures without the need for extensive sample preparation. Deteriorations of an API may be analyzed using this method, e.g., by comparing the diffraction pattern of the API to a known standard of the API prior to packaging.
Thermal methods of analysis may include, e.g., differential scanning calorimetry (DSC), thermogravimetry (TG), and hot-stage microscopy (HSM). All three methods provide information upon heating the sample. Depending on the information required, heating can be static or dynamic in nature.
Differential scanning calorimetry monitors the energy required to maintain the sample and a reference at the same temperature as they are heated. A plot of heat flow (W/g or J/g) versus temperature is obtained. The area under a DSC peak is directly proportional to the heat absorbed or released and integration of the peak results in the heat of transition.
Thermogravimetry (TG) measures the weight change of a sample as a function of temperature. A total volatile content of the sample is obtained, but no information on the identity of the evolved gas is provided. The evolved gas must be identified by other methods, such as gas chromatography, Karl Fisher titration (specifically to measure water), TG-mass spectroscopy, or TG-infrared spectroscopy. The temperature of the volatilization and the presence of steps in the TG curve can provide information on how tightly water or solvent is held in the lattice. If the temperature of the TG volatilization is similar to an endothermic peak in the DSC, the DSC peak is likely due or partially due to volatilization. It may be necessary to utilize multiple techniques to determine if more than one thermal event is responsible for a given DSC peak.
Hot-Stage Microscopy (HSM) is a technique that supplements DSC and TG. Events observed by DSC and/or TG can be readily characterized by HSM. Melting, gas evolution, and solid-solid transformations can be visualized, providing the most straightforward means of identifying thermal events. Thermal analysis can be used to determine the melting points, recrystallizations, solid-state transformations, decompositions, and volatile contents of pharmaceutical materials.
Other methods to analyze degradation or alteration of API and excipients are infrared (IR) and Raman spectroscopy. These techniques are sensitive to the structure, conformation, and environment of organic compounds. Infrared spectroscopy is based on the conversion of IR radiation into molecular vibrations. For a vibration to be IR-active, it must involve a changing molecular dipole (asymmetric mode). For example, vibration of a dipolar carbonyl group is detectable by IR spectroscopy. Whereas IR has been traditionally used as an aid in structure elucidation, vibrational changes also serve as probes of intermolecular interactions in solid materials.
Raman spectroscopy is based on the inelastic scattering of laser radiation with loss of vibrational energy by a sample. A vibrational mode is Raman active when there is a change in the polarizability during the vibration. Symmetric modes tend to be Raman-active. For example, vibrations about bonds between the same atom, such as in alkynes, can be observed by Raman spectroscopy.
NMR spectroscopy probes atomic environments based on the different resonance frequencies exhibited by nuclei in a strong magnetic field. Many different nuclei are observable by the NMR technique, but those of hydrogen and carbon atoms are most frequently studied. Solid-state NMR measurements are extremely useful for characterizing the crystal forms of pharmaceutical solids. Nuclei that are typically analyzed with this technique include those of 13C, 31P, 15N, 25Mg, and 23Na.
Chromatography is a general term applied to a wide variety of separation techniques based on the sample partitioning between a moving phase, which can be a gas, liquid, or supercritical fluid, and a stationary phase, which may be either a liquid or a solid. Generally, the crux of chromatography lies in the highly selective chemical interactions that occur in both the mobile and stationary phases. For example, depending on the API and the separation required, one or more of absorption, ion-exchange, size-exclusion, bonded phase, reverse, or normal phase stationary phases may be employed.
Mass spectrometry (MS) is an analytical technique that works by ionizing chemical compounds to generate charged molecules or molecule fragments and measuring their mass-to-charge ratios. Based on this analysis method, one can determine, e.g., the isotopic composition of elements in an API and determine the structure of the API by observing its fragmentation pattern.
It would be understood that the foregoing methods do not represent a comprehensive list of means by which one can analyze possible deteriorations, alterations, or concentrations of certain APIs. Therefore, it would be understood that other methods for determining the physical amounts and/or characteristics of an API may be employed. Additional methods may include, but are not limited to, e.g., Capillary Electrophoresis (CE), Atomic Spectroscopy (AS), and Luminescence Spectroscopy (LS).
EXAMPLES
The embodiments of the delamination resistant pharmaceutical containers described herein will be further clarified by the following examples.
Example 1
Six exemplary inventive glass compositions (compositions A-F) were prepared. The specific compositions of each exemplary glass composition are reported below in Table 8. Multiple samples of each exemplary glass composition were produced. One set of samples of each composition was ion exchanged in a molten salt bath of 100% KNO3 at a temperature of 450° C. for at least 5 hours to induce a compressive layer in the surface of the sample. The compressive layer had a surface compressive stress of at least 500 MPa and a depth of layer of at least 45 μm.
The chemical durability of each exemplary glass composition was then determined utilizing the DIN 12116 standard, the ISO 695 standard, and the ISO 720 standard described above. Specifically, non-ion exchanged test samples of each exemplary glass composition were subjected to testing according to one of the DIN 12116 standard, the ISO 695 standard, or the ISO 720 standard to determine the acid resistance, the base resistance or the hydrolytic resistance of the test sample, respectively. The hydrolytic resistance of the ion exchanged samples of each exemplary composition was determined according to the ISO 720 standard. The average results of all samples tested are reported below in Table 8.
As shown in Table 8, exemplary glass compositions A-F all demonstrated a glass mass loss of less than 5 mg/dm2 and greater than 1 mg/dm2 following testing according to the DIN 12116 standard with exemplary glass composition E having the lowest glass mass loss at 1.2 mg/dm2. Accordingly, each of the exemplary glass compositions were classified in at least class S3 of the DIN 12116 standard, with exemplary glass composition E classified in class S2. Based on these test results, it is believed that the acid resistance of the glass samples improves with increased SiO2 content.
Further, exemplary glass compositions A-F all demonstrated a glass mass loss of less than 80 mg/dm2 following testing according to the ISO 695 standard with exemplary glass composition A having the lowest glass mass loss at 60 mg/dm2. Accordingly, each of the exemplary glass compositions were classified in at least class A2 of the ISO 695 standard, with exemplary glass compositions A, B, D and F classified in class A1. In general, compositions with higher silica content exhibited lower base resistance and compositions with higher alkali/alkaline earth content exhibited greater base resistance.
Table 8 also shows that the non-ion exchanged test samples of exemplary glass compositions A-F all demonstrated a hydrolytic resistance of at least Type HGA2 following testing according to the ISO 720 standard with exemplary glass compositions C-F having a hydrolytic resistance of Type HGA1. The hydrolytic resistance of exemplary glass compositions C-F is believed to be due to higher amounts of SiO2 and the lower amounts of Na2O in the glass compositions relative to exemplary glass compositions A and B.
Moreover, the ion exchanged test samples of exemplary glass compositions B-F demonstrated lower amounts of extracted Na2O per gram of glass than the non-ion exchanged test samples of the same exemplary glass compositions following testing according to the ISO 720 standard.







TABLE 8







Composition and Properties of Exemplary Glass Compositions









Composition in mole %














A
B
C
D
E
F
















SiO2
70.8
72.8
74.8
76.8
76.8
77.4


Al2O3
7.5
7
6.5
6
6
7


Na2O
13.7
12.7
11.7
10.7
11.6
10


K2O
1
1
1
1
0.1
0.1


MgO
6.3
5.8
5.3
4.8
4.8
4.8


CaO
0.5
0.5
0.5
0.5
0.5
0.5


SnO2
0.2
0.2
0.2
0.2
0.2
0.2


DIN 12116
3.2
2.0
1.7
1.6
1.2
1.7


(mg/dm2)








classification
S3
S3
S3
S3
S2
S3


ISO 695
60.7
65.4
77.9
71.5
76.5
62.4


(mg/dm2)








classification
A1
A1
A2
A1
A2
A1


ISO 720
100.7
87.0
54.8
57.5
50.7
37.7


(ug Na2O/








g glass)








classification
HGA2
HGA2
HGA1
HGA1
HGA1
HGA1


ISO 720
60.3
51.9
39.0
30.1
32.9
23.3


(with IX)








(ug Na2O/








g glass)








classification
HGA1
HGA1
HGA1
HGA1
HGA1
HGA1









Example 2
Three exemplary inventive glass compositions (compositions G-I) and three comparative glass compositions (compositions 1-3) were prepared. The ratio of alkali oxides to alumina (i.e., Y:X) was varied in each of the compositions in order to assess the effect of this ratio on various properties of the resultant glass melt and glass. The specific compositions of each of the exemplary inventive glass compositions and the comparative glass compositions are reported in Table 9. The strain point, anneal point, and softening point of melts formed from each of the glass compositions were determined and are reported in Table 2. In addition, the coefficient of thermal expansion (CTE), density, and stress optic coefficient (SOC) of the resultant glasses were also determined and are reported in Table 9. The hydrolytic resistance of glass samples formed from each exemplary inventive glass composition and each comparative glass composition was determined according to the ISO 720 Standard both before ion exchange and after ion exchange in a molten salt bath of 100% KNO3 at 450° C. for 5 hours. For those samples that were ion exchanged, the compressive stress was determined with a fundamental stress meter (FSM) instrument, with the compressive stress value based on the measured stress optical coefficient (SOC). The FSM instrument couples light into and out of the birefringent glass surface. The measured birefringence is then related to stress through a material constant, the stress-optic or photoelastic coefficient (SOC or PEC) and two parameters are obtained: the maximum surface compressive stress (CS) and the exchanged depth of layer (DOL). The diffusivity of the alkali ions in the glass and the change in stress per square root of time were also determined







TABLE 9







Glass properties as a function of alkali to alumina ratio









Composition Mole %














G
H
I
1
2
3
















SiO2
76.965
76.852
76.962
76.919
76.960
77.156


Al2O3
5.943
6.974
7.958
8.950
4.977
3.997


Na2O
11.427
10.473
9.451
8.468
12.393
13.277


K2O
0.101
0.100
0.102
0.105
0.100
0.100


MgO
4.842
4.878
4.802
4.836
4.852
4.757


CaO
0.474
0.478
0.481
0.480
0.468
0.462


SnO2
0.198
0.195
0.197
0.197
0.196
0.196


Strain (° C.)
578
616
654
683
548
518


Anneal (° C.)
633
674
716
745
600
567


Softening (° C.)
892
946
1003
1042
846
798


Expansion (10−7 K−1)
67.3
64.3
59.3
55.1
71.8
74.6


Density (g/cm3)
2.388
2.384
2.381
2.382
2.392
2.396


SOC (nm/mm/Mpa)
3.127
3.181
3.195
3.232
3.066
3.038


ISO720 (non-IX)
88.4
60.9
47.3
38.4
117.1
208.1


ISO720 (IX450° C.-5 hr)
25.3
26
20.5
17.8
57.5
102.5


R2O/Al2O3
1.940
1.516
1.200
0.958
2.510
3.347


CS@t = 0 (MPa)
708
743
738
655
623
502


CS/√t (MPa/hr1/2)
−35
−24
−14
−7
−44
−37


D (μm2/hr)
52.0
53.2
50.3
45.1
51.1
52.4









The data in Table 9 indicates that the alkali to alumina ratio Y:X influences the melting behavior, hydrolytic resistance, and the compressive stress obtainable through ion exchange strengthening. In particular, FIG. 1 graphically depicts the strain point, anneal point, and softening point as a function of Y:X ratio for the glass compositions of Table 9. FIG. 1 demonstrates that, as the ratio of Y:X decreases below 0.9, the strain point, anneal point, and softening point of the glass rapidly increase. Accordingly, to obtain a glass which is readily meltable and formable, the ratio Y:X should be greater than or equal to 0.9 or even greater than or equal to 1.
Further, the data in Table 2 indicates that the diffusivity of the glass compositions generally decreases with the ratio of Y:X. Accordingly, to achieve glasses can be rapidly ion exchanged in order to reduce process times (and costs) the ratio of Y:X should be greater than or equal to 0.9 or even greater than or equal to 1.
Moreover, FIG. 2 indicates that for a given ion exchange time and ion exchange temperature, the maximum compressive stresses are obtained when the ratio of Y:X is greater than or equal to about 0.9, or even greater than or equal to about 1, and less than or equal to about 2, specifically greater than or equal to about 1.3 and less than or equal to about 2.0. Accordingly, the maximum improvement in the load bearing strength of the glass can be obtained when the ratio of Y:X is greater than about 1 and less than or equal to about 2. It is generally understood that the maximum stress achievable by ion exchange will decay with increasing ion-exchange duration as indicated by the stress change rate (i.e., the measured compressive stress divided by the square root of the ion exchange time). FIG. 2 generally shows that the stress change rate decreases as the ratio Y:X decreases.
FIG. 3 graphically depicts the hydrolytic resistance (y-axis) as a function of the ratio Y:X (x-axis). As shown in FIG. 3, the hydrolytic resistance of the glasses generally improves as the ratio Y:X decreases.
Based on the foregoing it should be understood that glasses with good melt behavior, superior ion exchange performance, and superior hydrolytic resistance can be achieved by maintaining the ratio Y:X in the glass from greater than or equal to about 0.9, or even greater than or equal to about 1, and less than or equal to about 2.
Example 3
Three exemplary inventive glass compositions (compositions J-L) and three comparative glass compositions (compositions 4-6) were prepared. The concentration of MgO and CaO in the glass compositions was varied to produce both MgO-rich compositions (i.e., compositions J-L and 4) and CaO-rich compositions (i.e., compositions 5-6). The relative amounts of MgO and CaO were also varied such that the glass compositions had different values for the ratio (CaO/(CaO+MgO)). The specific compositions of each of the exemplary inventive glass compositions and the comparative glass compositions are reported below in Table 10. The properties of each composition were determined as described above with respect to Example 2.







TABLE 10







Glass properties as function of CaO content









Composition Mole %














J
K
L
4
5
6
















SiO2
76.99
77.10
77.10
77.01
76.97
77.12


Al2O3
5.98
5.97
5.96
5.96
5.97
5.98


Na2O
11.38
11.33
11.37
11.38
11.40
11.34


K2O
0.10
0.10
0.10
0.10
0.10
0.10


MgO
5.23
4.79
3.78
2.83
1.84
0.09


CaO
0.07
0.45
1.45
2.46
3.47
5.12


SnO2
0.20
0.19
0.19
0.19
0.19
0.19


Strain (° C.)
585
579
568
562
566
561


Anneal (° C.)
641
634
620
612
611
610


Softening (° C.)
902
895
872
859
847
834


Expansion (10−7 K−1)
67.9
67.1
68.1
68.8
69.4
70.1


Density (g/cm3)
2.384
2.387
2.394
2.402
2.41
2.42


SOC nm/mm/Mpa
3.12
3.08
3.04
3.06
3.04
3.01


ISO720 (non-IX)
83.2
83.9
86
86
88.7
96.9


ISO720 (IX450° C.-5 hr)
29.1

28.4
33.2
37.3
40.1


Fraction of RO as CaO
0.014
0.086
0.277
0.465
0.654
0.982


CS@t = 0 (MPa)
707
717
713
689
693
676


CS/√t (MPa/hr1/2)
−36
−37
−39
−38
−43
−44


D (μm2/hr)
57.2
50.8
40.2
31.4
26.4
20.7









FIG. 4 graphically depicts the diffusivity D of the compositions listed in Table 10 as a function of the ratio (CaO/(CaO+MgO)). Specifically, FIG. 4 indicates that as the ratio (CaO/(CaO+MgO)) increases, the diffusivity of alkali ions in the resultant glass decreases thereby diminishing the ion exchange performance of the glass. This trend is supported by the data in Table 10 and FIG. 5. FIG. 5 graphically depicts the maximum compressive stress and stress change rate (y-axes) as a function of the ratio (CaO/(CaO+MgO)). FIG. 5 indicates that as the ratio (CaO/(CaO+MgO)) increases, the maximum obtainable compressive stress decreases for a given ion exchange temperature and ion exchange time. FIG. 5 also indicates that as the ratio (CaO/(CaO+MgO)) increases, the stress change rate increases (i.e., becomes more negative and less desirable).
Accordingly, based on the data in Table 10 and FIGS. 4 and 5, it should be understood that glasses with higher diffusivities can be produced by minimizing the ratio (CaO/(CaO+MgO)). It has been determined that glasses with suitable diffusivities can be produced when the (CaO/(CaO+MgO)) ratio is less than about 0.5. The diffusivity values of the glass when the (CaO/(CaO+MgO)) ratio is less than about 0.5 decreases the ion exchange process times needed to achieve a given compressive stress and depth of layer. Alternatively, glasses with higher diffusivities due to the ratio (CaO/(CaO+MgO)) may be used to achieve a higher compressive stress and depth of layer for a given ion exchange temperature and ion exchange time.
Moreover, the data in Table 10 also indicates that decreasing the ratio (CaO/(CaO+MgO)) by increasing the MgO concentration generally improves the resistance of the glass to hydrolytic degradation as measured by the ISO 720 standard.
Example 4
Three exemplary inventive glass compositions (compositions M-O) and three comparative glass compositions (compositions 7-9) were prepared. The concentration of B2O3 in the glass compositions was varied from 0 mol. % to about 4.6 mol. % such that the resultant glasses had different values for the ratio B2O3/(R2O−Al2O3). The specific compositions of each of the exemplary inventive glass compositions and the comparative glass compositions are reported below in Table 11. The properties of each glass composition were determined as described above with respect to Examples 2 and 3.







TABLE 11







Glass properties as a function of B2O3 content









Composition Mole %














M
N
O
7
8
9
















SiO2
76.860
76.778
76.396
74.780
73.843
72.782


Al2O3
5.964
5.948
5.919
5.793
5.720
5.867


B2O3
0.000
0.214
0.777
2.840
4.443
4.636


Na2O
11.486
11.408
11.294
11.036
10.580
11.099


K2O
0.101
0.100
0.100
0.098
0.088
0.098


MgO
4.849
4.827
4.801
4.754
4.645
4.817


CaO
0.492
0.480
0.475
0.463
0.453
0.465


SnO2
0.197
0.192
0.192
0.188
0.183
0.189


Strain (° C.)
579
575
572
560
552
548


Anneal (° C.)
632
626
622
606
597
590


Softening (° C.)
889
880
873
836
816
801


Expansion (10−7 K−1)
68.3
67.4
67.4
65.8
64.1
67.3


Density (g/cm3)
2.388
2.389
2.390
2.394
2.392
2.403


SOC (nm/mm/MPa)
3.13
3.12
3.13
3.17
3.21
3.18


ISO720 (non-IX)
86.3
78.8
68.5
64.4
52.7
54.1


ISO720 (IX450° C.-5 hr)
32.2
30.1
26
24.7
22.6
26.7


B2O3/(R2O—Al2O3)
0.000
0.038
0.142
0.532
0.898
0.870


CS@t = 0 (MPa)
703
714
722
701
686
734


CS/√t (MPa/hr1/2)
−38
−38
−38
−33
−32
−39


D (μm2/hr)
51.7
43.8
38.6
22.9
16.6
15.6









FIG. 6 graphically depicts the diffusivity D (y-axis) of the glass compositions in Table 11 as a function of the ratio B2O3/(R2O−Al2O3) (x-axis) for the glass compositions of Table 11. As shown in FIG. 6, the diffusivity of alkali ions in the glass generally decreases as the ratio B2O3/(R2O−Al2O3) increases.
FIG. 7 graphically depicts the hydrolytic resistance according to the ISO 720 standard (y-axis) as a function of the ratio B2O3/(R2O−Al2O3) (x-axis) for the glass compositions of Table 11. As shown in FIG. 6, the hydrolytic resistance of the glass compositions generally improves as the ratio B2O3/(R2O−Al2O3) increases.
Based on FIGS. 6 and 7, it should be understood that minimizing the ratio B2O3/(R2O−Al2O3) improves the diffusivity of alkali ions in the glass thereby improving the ion exchange characteristics of the glass. Further, increasing the ratio B2O3/(R2O−Al2O3) also generally improves the resistance of the glass to hydrolytic degradation. In addition, it has been found that the resistance of the glass to degradation in acidic solutions (as measured by the DIN 12116 standard) generally improves with decreasing concentrations of B2O3. Accordingly, it has been determined that maintaining the ratio B2O3/(R2O−Al2O3) to less than or equal to about 0.3 provides the glass with improved hydrolytic and acid resistances as well as providing for improved ion exchange characteristics.
It should now be understood that the glass compositions described herein exhibit chemical durability as well as mechanical durability following ion exchange. These properties make the glass compositions well suited for use in various applications including, without limitation, pharmaceutical packaging materials.
Example 5: Determining the Presence and Amount of Glass Flakes in Pharmaceutical Solutions
The resistance to delamination may be characterized by the number of glass particulates present in a pharmaceutical solution contained within a glass container described herein after. In order to assess the long-term resistance of the glass container to delamination, an accelerated delamination test is utilized. The test consists of washing the glass container at room temperature for 1 minute and depyrogenating the container at about 320° C. for 1 hour. Thereafter a pharmaceutical solution is placed in the glass container to 80-90% full, the glass container is closed, and rapidly heated to, for example, 100° C. and then heated from 100° C. to 121° C. at a ramp rate of 1 deg/min at a pressure of 2 atmospheres. The glass container and solution are held at this temperature for 60 minutes, cooled to room temperature at a rate of 0.5 deg/min and the heating cycle and hold are repeated. The glass container is then heated to 50° C. and held for two days for elevated temperature conditioning. After heating, the glass container is dropped from a distance of at least 18′′ onto a firm surface, such as a laminated tile floor, to dislodge any flakes or particles that are weakly adhered to the inner surface of the glass container.
Thereafter, the pharmaceutical solution contained in the glass container is analyzed to determine the number of glass particles present per liter of solution. Specifically, the solution from the glass container is directly poured onto the center of a Millipore Isopore Membrane filter (Millipore #ATTP02500 held in an assembly with parts #AP1002500 and #M000025A0) attached to vacuum suction to draw the solution through the filter within 10-15 seconds. Particulate flakes are then counted by differential interference contrast microscopy (DIC) in the reflection mode as described in “Differential interference contrast (DIC) microscopy and modulation contrast microscopy” from Fundamentals of light microscopy and digital imaging. New York: Wiley-Liss, pp 153-168. The field of view is set to approximately 1.5 mm×1.5 mm and particles larger than 50 microns are counted manually. There are 9 such measurements made in the center of each filter membrane in a 3×3 pattern with no overlap between images. A minimum of 100 mL of solution is tested. As such, the solution from a plurality of small containers may be pooled to bring the total amount of solution to 100 mL. If the containers contain more than 10 mL of solution, the entire amount of solution from the container is examined for the presence of particles. For containers having a volume greater than 10 mL containers, the test is repeated for a trial of 10 containers formed from the same glass composition under the same processing conditions and the result of the particle count is averaged for the 10 containers to determine an average particle count. Alternatively, in the case of small containers, the test is repeated for a trial of 10 sets of 10 mL of solution, each of which is analyzed and the particle count averaged over the 10 sets to determine an average particle count. Averaging the particle count over multiple containers accounts for potential variations in the delamination behavior of individual containers.
It should be understood that the aforementioned test is used to identify particles which are shed from the interior wall(s) of the glass container due to delamination and not tramp particles present in the container from forming processes. Specifically, delamination particles will be differentiated from tramp glass particles based on the aspect ratio of the particle (i.e., the ratio of the width of the particle to the thickness of the particle). Delamination produces particulate flakes or lamellae which are irregularly shaped and are typically >50 μm in diameter but often >200 μm. The thickness of the flakes is usually greater than about 100 nm and may be as large as about 1 μm. Thus, the minimum aspect ratio of the flakes is typically >50. The aspect ratio may be greater than 100 and sometimes greater than 1000. Particles resulting from delamination processes generally have an aspect ratio which is generally greater than about 50. In contrast, tramp glass particles will generally have a low aspect ratio which is less than about 3. Accordingly, particles resulting from delamination may be differentiated from tramp particles based on aspect ratio during observation with the microscope. Validation results can be accomplished by evaluating the heel region of the tested containers. Upon observation, evidence of skin corrosion/pitting/flake removal, as described in “Nondestructive Detection of Glass Vial Inner Surface Morphology with Differential Interference Contrast Microscopy” from Journal of Pharmaceutical Sciences 101(4), 2012, pages 1378-1384, is noted.
Using this method, pharmaceutical compositions can be tested for the presence of glass flakes and various compositions can be compared to each other to assess the safety of various pharmaceutical compositions.
Example 6: Stability Testing of Pharmaceutical Compositions
Stability studies are part of the testing required by the FDA and other regulatory agencies. Stability studies should include testing of those attributes of the API that are susceptible to change during storage and are likely to influence quality, safety, and/or efficacy. The testing should cover, as appropriate, the physical, chemical, biological, and microbiological attributes of the API (e.g., small molecule or biologic therapeutic agent) in the container with the closure to be used for storage of the agent. If the API is formulated as a liquid by the manufacturer, the final formulation should be assayed for stability. If the API is formulated as an agent for reconstitution by the end user using a solution provided by the manufacturer, both the API and the solution for reconstitution are preferably tested for stability as the separate packaged components (e.g., the API subjected to storage reconstituted with solution for reconstitution not subject to storage, API not subject to storage reconstituted with a solution subject to storage, and both API and solution subject to storage). This is particularly the case when the solution for reconstitution includes an active agent (e.g., an adjuvant for reconstitution of a vaccine).
In general, a substance API should be evaluated under storage conditions (with appropriate tolerances) that test its thermal stability and, if applicable, its sensitivity to moisture. The storage conditions and the lengths of studies chosen should be sufficient to cover storage, shipment, and subsequent use.
API should be stored in the container(s) in which the API will be provided to the end user (e.g., vials, ampules, syringes, injectable devices). Stability testing methods provided herein refer to samples being removed from the storage or stress conditions indicated. Removal of a sample preferably refers to removing an entire container from the storage or stress conditions. Removal of a sample should not be understood as withdrawing a portion of the API from the container as removal of a portion of the API from the container would result in changes of fill volume, gas environment, etc. At the time of testing the API subject to stability and/or stress testing, portions of the samples subject to stability and/or stress testing can be used for individual assays.
The long-term testing should cover a minimum of 12 months' duration on at least three primary batches at the time of submission and should be continued for a period of time sufficient to cover the proposed retest period. Additional data accumulated during the assessment period of the registration application should be submitted to the authorities if requested. Data from the accelerated storage condition and, if appropriate, from the intermediate storage condition can be used to evaluate the effect of short-term excursions outside the label storage conditions (such as might occur during shipping).
Long-term, accelerated, and, where appropriate, intermediate storage conditions for API are detailed in the sections below. The general case should apply if the API is not specifically covered by a subsequent section. It is understood that the time points for analysis indicated in the table are suggested end points for analysis. Interim analysis can be performed at shorter time points (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or 11 months). For API to be labeled as stable for storage for more than 12 months, time points beyond 12 months can be assessed (e.g., 15, 18, 21, 24 months). Alternative storage conditions can be used if justified.







TABLE 12







General Conditions for Stability Analysis













Time points



Study
Storage condition
for analysis







Long-term
Long-term* 25° C. ± 2° C./
12 months 




60% RH ± 5% RH or




30° C. ± 2° C./65% RH ± 5% RH



Intermediate
30° C. ± 2° C./65% RH ± 5% RH
6 months



Accelerated
40° C. ± 2° C./75% RH ± 5% RH
6 months

















TABLE 13







Conditions for Stability Analysis for Storage in a Refrigerator











Minimum time period




covered by data at


Study
Storage condition
submission





Long-term
 5° C. ± 3° C.
12 months


Accelerated
25° C. ± 2° C./60% RH ± 5% RH
 6 months
















TABLE 14







Conditions for Stability Analysis for Storage in a Freezer













Minimum time period





covered by data at



Study
Storage condition
submission







Long-term
−20° C. ± 5° C.
12 months










Storage condition for API intended to be stored in a freezer, testing on a single batch at an elevated temperature (e.g., 5° C.±3° C. or 25° C.±2° C.) for an appropriate time period should be conducted to address the effect of short-term excursions outside the proposed label storage condition (e.g., stress during shipping or handling, e.g., increased temperature, multiple freeze-thaw cycles, storage in a non-upright orientation, shaking, etc.).
The assays performed to assess stability of an API include assays to that are used across most APIs to assess the physical properties of the API, e.g., degradation, pH, color, particulate formation, concentration, toxicity, etc. Assays to detect the general properties of the API are also selected based on the chemical class of the agent, e.g., denaturation and aggregation of protein based API. Assays to detect the potency of the API, i.e., the ability of the API to achieve its intended effect as demonstrated by the quantitative measurement of an attribute indicative of the clinical effect as compared to an appropriate control, are selected based on the activity of the particular agent. For example, the biological activity of the API, e.g., enzyme inhibitor activity, cell killing activity, anti-inflammatory activity, coagulation modulating activity, etc., is measured using in vitro and/or in vivo assays such as those provided herein. Pharmacokinetic and toxicological properties of the API are also assessed using methods known in the art, such as those provided herein.
Example 7: Analysis of Adherence to Glass Vials
Changes in the surface of glass can result in changes in the adherence of API to glass. The amount of agent in samples withdrawn from glass vials are tested at intervals to determine if the concentration of the API in solution changes over time. API are incubated in containers as described in the stability testing and/or stress testing methods provided in Example 6. Preferably, the API is incubated both in standard glass vials with appropriate closures and glass vials such as those provided herein. At the desired intervals, samples are removed and assayed to determine the concentration of the API in solution. The concentration of the API is determined using methods and controls appropriate to the API. The concentration of the API is preferably determined in conjunction with at least one assay to confirm that the API, rather than degradation products of the API, is detected. In the case of biologics in which the conformational structure of the biologic agent is essential to its function of the API, the assays for concentration of the biologic are preferably preformed in conjunction with an assay to confirm the structure of the biologic (e.g., activity assay).
For example, in the cases of small molecule APIs, the amount of agent present is determined, for example, by mass spectrometry, optionally in combination with liquid chromatography, as appropriate, to separate the agent from any degradation products that may be present in the sample.
For protein based biologic APIs, the concentration of the API is determined, for example, using ELISA assay. Chromatography methods are used in conjunction with methods to determine protein concentration to confirm that protein fragments or aggregates are not being detected by the ELISA assay.
For nucleic acid biologic APIs, the concentration of the API is determined, for example, using quantitative PCR when the nucleic acids are of sufficient length to permit detection by such methods. Chromatography methods are used to determine both the concentration and size of nucleic acid based API.
For viral vaccine APIs, the concentration of the virus is determined, for example, using colony formation assays.
Example 8: Analysis of Pharmacokinetic Properties
Pharmacokinetics is concerned with the analysis of absorption, distribution, metabolism, and excretion of API. Storage and stress can potentially affect the pharmacokinetic properties of various API. To assess pharmacokinetics of API subject to stability and/or stress testing, agents are incubated in containers as described in Example 6. Preferably, the API are incubated both in standard glass vials with appropriate closures and glass vials such as those provided herein. At the desired intervals, samples are removed and assayed.
The API is delivered to subjects by the typical route of delivery for the API (e.g., injection, oral, topical). As pharmacokinetics are concerned with the absorption and elimination of the API, normal subjects are typically used to assess pharmacokinetic properties of the API. However, if the API is to be used in subjects with compromised ability to absorb or eliminate the API (e.g., subjects with liver or kidney disease), testing in an appropriate disease model may be advantageous. Depending on the half-life of the compound, samples (e.g., blood, urine, stool) are collected at predetermined time points (e.g., 0 min, 30 min, 60 min, 90 min, 120 min, 4 hours, 6 hours, 12 hours, 24 hours, 36 hours, 48 hours, etc.) for at least two, preferably three half-lives of the API, and analyzed for the presence of the API and metabolic products of the API. At the end of the study, organs are harvested and analyzed for the presence of the API and metabolic products of the API.
The results are analyzed using an appropriate model selected based on, at least, the route of administration of the API. The pharmacokinetic properties of the API subjected to stability and/or stress testing are compared to API not subjected to stability or stress testing and other appropriate controls (e.g., vehicle control). Changes, if any, in pharmacokinetic properties as a result of storage of the API under each condition are determined.
Example 9: Analysis of Toxicity Profiles
Storage of API can result in alterations of toxicity of API as a result of reactivity of the API with the container, leeching of agents from the container, delamination resulting in particulates in the agent, reaction of the API molecules with each other or components of the storage buffer, or other causes.
Agents are incubated in containers as described in the stability testing and/or stress testing methods provided in Example 6. Preferably, the API is incubated both in standard glass vials with appropriate closures and glass vials such as those provided herein. At the desired intervals, samples are removed and assayed to determine the toxicity the API. The toxicity of the API is determined using methods and controls appropriate to the API. In vitro and in vivo testing can be used alone or in combination to assess changes in toxicity of agents as a result of storage or stress.
In in vitro assays, cell lines are grown in culture and contacted with increasing concentrations of API subjected to stability and/or stress testing for predetermined amounts of time (e.g., 12, 24, 36, 48, and 72 hours). Cell viability is assessed using any of a number of routine or commercially available assays. Cells are observed, for example, by microscopy or using fluorescence activated cell sorting (FACS) analysis using commercially available reagents and kits. For example, membrane-permeant calcein AM is cleaved by esterases in live cells to yield cytoplasmic green fluorescence, and membrane-impermeant ethidium homodimer-1 labels nucleic acids of membrane-compromised cells with red fluorescence. Membrane-permeant SYTO 10 dye labels the nucleic acids of live cells with green fluorescence, and membrane-impermeant DEAD Red dye labels nucleic acids of membrane-compromised cells with red fluorescence. A change in the level of cell viability is detected between the cells contacted with API subjected to stress and/or stability testing in standard glass vials as compared to the glass vials provided herein and appropriate controls (e.g., API not subject to stability testing, vehicle control).
In vivo toxicity assays are performed in animals. Typically preliminary assays are performed on normal subjects. However, if the disease or condition to be treated could alter the susceptibility of the subject to toxic agents (e.g., decreased liver function, decreased kidney function), toxicity testing in an appropriate model of the disease or condition can be advantageous. One or more doses of agents subjected to stability and/or stress testing are administered to animals. Typically, doses are far higher (e.g., 5 times, 10 times) the dose that would be used therapeutically and are selected, at least in part, on the toxicity of the API not subject to stability and/or stress testing. However, for the purpose of assaying stability of API, the agent can be administered at a single dose that is close to (e.g., 70%-90%), but not at, a dose that would be toxic for the API not subject to stability or stress testing. In single dose studies, after administration of the API subject to stress and/or stability testing (e.g., 12 hours, 24 hours, 48 hours, 72 hours), during which time blood, urine, and stool samples may be collected. In long term studies, animals are administered a lower dose, closer to the dose used for therapeutic treatment, and are observed for changes indicating toxicity, e.g., weight loss, loss of appetite, physical changes, or death. In both short and long term studies, organs are harvested and analyzed to determine if the API is toxic. Organs of most interest are those involved in clearance of the API, e.g., liver and kidneys, and those for which toxicity would be most catastrophic, e.g., heart, brain. An analysis is performed to detect a change in toxicity between the API subjected to stress and/or stability testing in standard glass vials as compared to the glass vials provided herein, as compared to API not subject to stability and/or stress testing and vehicle control. Changes, if any, in toxicity properties as a result of storage of the API under each condition are determined.
Example 10: Analysis of Pharmacodynamic Profiles
Pharmacodynamics includes the study of the biochemical and physiological effects of drugs on the body or on microorganisms or parasites within or on the body and the mechanisms of drug action and the relationship between drug concentration and effect. Mouse models for a large variety of disease states are known and commercially available (see, e.g., from the Jackson Laboratory). A number of induced models of disease are also known.
Agents are incubated in containers as described in the stability testing and/or stress testing methods provided in Example 6. Preferably, the samples are incubated both in standard glass vials with appropriate closures and glass vials such as those provided herein. At the desired intervals, samples are removed and assayed for pharmacodynamic activity using known animal models. Exemplary mouse models for testing the various classes of agents indicated are known in the art.
The mouse is treated with the API subject to stability and/or stress testing. The efficacy of the API subject to stability and/or stress testing to treat the appropriate disease or condition is assayed as compared to API not subject to stability and/or stress testing and vehicle control. Changes, if any, in pharmacodynamic properties as a result of storage of the API under each condition are determined.
Example 11: Confirmation of Stability and Activity of PEDIARIX® (Diphtheria and Tetanus Toxoids and Acellular Pertussis Adsorbed, Hepatitis B (Recombinant) and Inactivated Poliovirus Vaccine)
Diphtheria and Tetanus Toxoids and Acellular Pertussis Adsorbed, Hepatitis B (Recombinant) and Inactivated Poliovirus Vaccine (PEDIARIX®) is a combination of diphtheria toxoid, tetanus toxoid, inactivated pertussis toxin, hepatitis B virus surface antigen, inactivated Type I poliovirus (Mahoney), Type 2 poliovirus (MEF-1), Type 3 poliovirus (Saukett), filamentous hemagglutinin and pertactin (69 kD outer membrane protein). Protection against disease results from the development of neutralizing antibodies to the diphtheria toxin, the tetanus toxin, and to the poliovirus. The role of the B. pertussis components (inactivated pertussis toxin, filamentous hemagglutinin and pertactin (69 kD outer membrane protein)) in the development of immunity to pertussis is not well understood. Antibody concentrations >10 mIU/mL against HBsAg confer protection against hepatitis B virus infection. The diphtheria and tetanus toxins are produced in culture and detoxified with formaldehyde. The pertussis antigens are produced in culture and the toxin is inactivated using glutaraldehye and formaldehye and the filamentous hemagglutinin and pertactin are treated with formaldehyde. The hepatitis B surface antigen is obtained from cultured cells and purified by precipitation, ion exchange chromatography and ultrafiltration. The three strains of poliovirus are grown individually in cell culture, purified, inactivated using formaldehyde then pooled to form a trivalent concentrate.
PEDIARIX® is a noninfectious, sterile vaccine for intramuscular administration. Three 0.5 ml doses are administered at 2, 4 and 6 months of age. Each 0.5-ml dose contains 25 Lf of diphtheria toxoid, 10 Lf of tetanus toxoid, 25 mcg of inactivated pertussis toxin (PT), 25 mcg of filamentous hemagglutinin (FHA), 8 mcg of pertactin (69 kDa outer membrane protein), 10 mcg of HBsAg, 40 D-antigen Units (DU) of Type 1 poliovirus (Mahoney), 8 DU of Type 2 poliovirus (MEF-1), and 32 DU of Type 3 poliovirus (Saukett). PEDIARIX® is a suspension for injection available in 0.5 ml single-dose disposable prefilled syringe.
Diphtheria and Tetanus Toxoids and Acellular Pertussis Adsorbed, Hepatitis B (Recombinant) and Inactivated Poliovirus Vaccine samples are incubated in containers as described in the stability testing and/or stress testing methods provided in Example 6. Preferably, the samples are incubated both in standard glass vials with appropriate closures and glass vials such as those provided herein. At the desired intervals, samples are removed and assayed to determine the stability and/or activity of the agent. The activity of Diphtheria and Tetanus Toxoids and Acellular Pertussis Adsorbed, Hepatitis B (Recombinant) and Inactivated Poliovirus Vaccine is determined using methods and controls appropriate to the agent, e.g., using the methods provided in Podda et al., Vaccine 9, 741-745; Gustafsson et al., N Engl J Med 334, 349-355 (1996); Gillenius et al., J Biol Stand 13, 61-66 (1985); WO91/12020; WO98/00167; WO99/13906; US Publications 20110195087 and 20110206726; and U.S. Pat. No. 8,007,818; each of which is incorporated herein by reference.
In Vivo Immunogenicity Assay—Diphtheria Toxin
The in vivo immunogenicity of the diphtheria toxin is analyzed in guinea pigs. A 1.0 ml dose of the vaccine subject to stability and/or stress testing (i.e., test vaccine) is administered subcutaneously to each of 48 animals. An additional 48 animals are administered 1.0 ml dose of vaccine not subject to stability and/or stress testing (i.e., reference vaccine). The potency of the diphtheria toxin is determined by lethal challenge method whereby 3 dilutions of each of the test and the reference vaccine are prepared. The middle dilutions contain the ED50 dose that saves at least, or more than, 50% of the test animals. Sixteen guinea pigs are treated for every dilution of test vaccine and reference vaccine. An additional 20 guinea pigs are not immunized and used for the titration of the diphtheria toxin, with 5 guinea pigs used for each of 4 dilutions. After 28 days the test animals are challenged with a subcutaneous dose of diphtheria toxin containing 100 LD50. The sample passes the diphtheria potency test if it contains ≧30 I.U./Single Human Dose. The test vaccine should fulfill linearity and parallelism with reference vaccine. The fiducial limit of estimated potency should be between 50% and 200%. The estimated potency should not be less than 30 I.U. per single human dose. The 95% confidence interval of estimate of potency should be within 50% and 200% unless the lower limit of the 95% confidence interval of the estimated potency should be ≧30 I.U. per dose.
In Vivo Immunogenicity Assay—Tetanus Toxin
The in vivo immunogenicity of the tetanus toxin is analyzed in Swiss albino mice. A 0.5 ml dose of the vaccine subject to stability and/or stress testing (i.e., test vaccine) is administered subcutaneously to each of 48 animals. An additional 48 animals are administered and 0.5 ml dose of vaccine not subject to stability and/or stress testing (i.e., reference vaccine). The potency of the tetanus toxin is determined by lethal challenge method whereby 3 dilutions of each of the test and the reference vaccine are prepared. The middle dilutions contain the ED50 dose that saves at least, or more than, 50% of the test animals. Sixteen mice are treated for every dilution of test vaccine and reference vaccine. An additional 20 mice are not immunized and used for the titration of the tetanus toxin, with 5 mice used for each of 4 dilutions. After 28 days the test animals are challenged with a subcutaneous dose of tetanus toxin containing 100 LD50. The sample passes the tetanus potency test if it contains ≧60 I.U./Single Human Dose. The test vaccine should fulfill linearity and parallelism with reference vaccine. The fiducial limit of estimated potency should be between 50% and 200%. The estimated potency should not be less than 60 I.U. per single human dose. The 95% confidence interval of estimate of potency should be within 50-200% unless the lower limit of the 95% confidence interval of the estimated potency should be ≧60 I.U. per dose.
ELISA to Detect Antibody Production—Pertussis Toxin, Filamentous Hemagglutinin and Pertactin
The antigenic effect of the pertussis toxin, filamentous hemagglutinin and pertactin is determined in Swiss mice by an ELISA method. Three dilutions (neat, 1:5 and 1:25) of the vaccine subject to stability and/or stress testing (i.e., test vaccine) and vaccine not subject to stability and/or stress testing (i.e., reference vaccine) are prepared. Six groups of 8 Swiss mice are immunized using 0.5 ml of one of each dilution by subcutaneous injection. Blood is collected 35 days after immunization. Serum samples are tested for antibodies against the pertussis antigens by ELISA. The test vaccine passes the potency test if ≧70% of the mice receiving the test vaccine are seroconverted.
In Vivo Immunogenicity Study—Pertussis Toxin, Filamentous Hemagglutinin and Pertactin
The in vivo immunogenicity of the pertussis toxin, filamentous hemagglutinin and pertactin is analyzed in twelve healthy infants who receive three doses of vaccine subject to stability and/or stress testing (i.e., test vaccine) and vaccine not subject to stability and/or stress testing (i.e., reference vaccine) by intramuscular injection at 2, 4 and 6 months of age. One month after the third dose of test or reference vaccine, the presence of pertussis toxin neutralizing antibodies are determined by the Chinese hamster ovary (CHO) cell assay using U.S. reference human pertussis antiserum containing 640 neutralizing units. Two-fold dilutions of 25 μl of antiserum obtained from infants immunized with test or reference vaccine, and reference antiserum, are added to culture microplate wells along with 25 μl of pertussis toxin containing 4 times the clustering concentration and incubated for 3 hours at 37° C. Thereafter, 10,000 trypsinized CHO cells and 200 μl of culture medium are added to each well and incubated for 48 hours. Clustering is determined by direct microscopic examination and is used to measure the antibody response to the vaccine. Neutralizing titers are expressed as the reciprocal of the highest serum dilution causing complete inhibition of the clustering activity induced by native toxin.
One month after the third dose of test or reference vaccine, levels of IgG and IgA to the antigens is determined by ELISA. U.S. reference human pertussis antiserum containing 200 EU/ml of IgG anti-pertussis toxin and 200 EU/ml IgG anti-filamentous hemagglutinin is used as a reference standard. An immune serum was assigned a value of 20 EU/ml of IgG anti-pertactin.
ELISA to Detect Antibody Production—Hepatitis B Surface Antigen
The antigenic effect of the hepatitis B surface antigen is determined in Balb/c mice using 5, two-fold dilutions each of vaccine subject to stability and/or stress testing (i.e., test vaccine) and vaccine not subject to stability and/or stress testing (i.e., reference vaccine). A 1.0 ml dose of test or reference vaccine is administered intraperitoneally to 10 mice per dilution. Ten mice are inoculated with diluent to serve as a placebo. Twenty-eight days after inoculation, blood is collected from the mice and the sera separated. The serum samples are tested for antibody titer against hepatitis B surface antigen using ELISA. The test vaccine passes the hepatitis B potency test if the upper limit of its relative potency is ≧1.
Antibody Production—Inactivated Polio Virus
The antigenic effect of the inactivated polio virus is analyzed in Wistar rats using 5, three-fold dilutions each of vaccine subject to stability and/or stress testing (i.e., test vaccine) and vaccine not subject to stability and/or stress testing (i.e., reference vaccine). A 0.5 ml dose of test or reference vaccine is administered intramuscularly to 10 rats per dilution. Twenty-one days after inoculation, blood is collected from the rats and the serum separated. The serum samples are tested for antibody titer against type 1, type 2, and type 3 serotypes of polio virus by serum neutralization test. The test is valid if the median effective dose (ED50) for both the test and reference vaccines lies between the smallest and the largest dose given to the animals, the statistical analysis shows no significant deviation from linearity or parallelism, and the fiducial limits of the estimated relative potency fall between 25% and 400% of the estimated potency.
Example 12: Confirmation of Stability and Activity of HAVRIX® (Hepatitis A Vaccine), ENGERIX-B® (Hepatitis B Vaccine (Recombinant)), and TWINRIX® (Hepatitis A & Hepatitis B (Recombinant) Vaccine)
Hepatitis A Vaccine (HAVRIX®) is a form of sterile suspension of inactivated hepatitis A virus (strain HM175). The presence of antibodies to hepatitis A virus confers protection against hepatitis A disease. The hepatitis A virus is propagated in MRC-5 human diploid cells and inactivated with formalin. Viral antigen activity is referenced to a standard using an enzyme linked immunosorbent assay (ELISA) and is expressed in terms of ELISA Units.
HAVRIX® is a sterile vaccine for intramuscular administration. Adults receive a single 1 ml dose and a 1 ml booster dose between 6 to 12 months later. Children and adolescents receive a single 0.5 ml dose and a 0.5 ml booster dose between 6 to 12 months later. Each 1 ml dose contains 1440 ELISA Units of viral antigen adsorbed onto 0.5 mg of aluminum as aluminum hydroxide. Each 0.5 ml dose contains 720 ELISA Units of viral antigen adsorbed onto 0.25 mg of aluminum as aluminum hydroxide. The dose also contains amino acid supplement (0.3% w/v) in a phosphate-buffered saline solution and polysorbate 20 (0.05 mg/ml). Residual compounds from the manufacturing process include not more than 5 mcg/ml residual MRC-5 cellular sulfate, not more than 0.1 mg/ml formalin and not more than 40 ng/ml neomycin sulfate. HAVRIX® is currently supplied in either a 0.5 ml or 1 ml single-dose vial and in a 0.5 ml or 1 ml prefilled syringe. HAVRIX® is a homogeneous turbid white suspension for injection.
Hepatitis B Vaccine (Recombinant) (ENGERIX-B®) is a form of sterile suspension of noninfectious hepatitis B virus surface antigen (HBsAg). Protection against hepatitis B virus infection is due to antibody concentrations >10 mIU/mL against HBsAg. Seroconversion is defined as antibody titers ≧1 mIU/ml. The hepatitis B surface antigen is derived from recombinant Saccharomyces cerevisiae cells which carry the surface antigen gene of the hepatitis B virus. The surface antigen is purified by precipitation, ion exchange chromatography and ultrafiltration.
ENGERIX-B® is a sterile vaccine for intramuscular administration. Individuals 20 years of age and older receive a series of 3, 1 ml doses given on a 0, 1, and 6 month schedule. Individuals from birth to 19 years of age receive a series of 3, 0.5 ml doses given on a 0, 1, and 6 month schedule. Each 1 ml dose contains 20 mcg of HBsAg adsorbed on 0.5 mg aluminum as aluminum hydroxide. Each 0.5 ml dose contains 10 mcg of HBsAg adsorbed on 0.25 mg aluminum as aluminum hydroxide. Excipients include sodium chloride (9 mg/ml) and phosphate buffers (disodium phosphate dihydrate, 0.98 mg/ml; sodium dihydrogen phosphate dihydrate, 0.71 mg/ml). Each dose contains no more than 5% yeast protein. ENGERIX® is currently supplied in either a 0.5 ml or 1 ml single-dose vial and in a 0.5 ml or 1 ml prefilled syringe. ENGERIX® is a homogeneous turbid white suspension for injection.
Hepatitis A & Hepatitis B (Recombinant) Vaccine (TWINRIX®) is a bivalent vaccine containing inactivated hepatitis A virus (strain HM175) and noninfectious hepatitis B virus surface antigen (HBsAg). The presence of antibodies to hepatitis A virus confers protection against hepatitis A disease. Protection against hepatitis B virus infection is due to antibody concentrations >10 mIU/mL against HBsAg. The hepatitis A virus is propagated in cell culture and inactivated with formalin. The hepatitis B surface antigen is obtained from cultured cells and purified by precipitation, ion exchange chromatography and ultrafiltration.
TWINRIX® is a sterile vaccine for intramuscular administration. Individuals receive a standard dosing regimen of 3, 1 ml doses at 0, 1, and 6 months. Under an accelerated dosing regimen, individuals receive 4, 1 ml doses on days 0, 7, and 21 to 30 followed by a booster dose at 12 months. Each 1 ml dose contains 720 ELISA Units of inactivated hepatitis A virus and 20 mcg of recombinant HBsAg protein. The 1 ml dose of vaccine also contains 0.45 mg of aluminum in the form of aluminum phosphate and aluminum hydroxide as adjuvants, amino acids, sodium chloride, phosphate buffer, polysorbate 20 and Water for Injection. TWINRIX® is currently supplied as a 1 ml single-dose vial and as a prefilled syringe and is formulated as a turbid white suspension for injection.
Hepatitis A Vaccine, Hepatitis B Vaccine (Recombinant), and a formulation including both vaccines, are incubated in containers as described in the stability testing and/or stress testing methods provided in Example 6. Preferably, the samples are incubated both in standard glass vials with appropriate closures and glass vials such as those provided herein. At the desired intervals, samples are removed and assayed to determine the stability and/or activity of the agent. The activity of Hepatitis A Vaccine, Hepatitis B Vaccine (Recombinant), and a formulation including both vaccines, is determined using methods and controls appropriate to the agent, e.g., using the methods provided in EP1073462B1; US Publication 20110195087; U.S. Pat. No. 5,151,023; and U.S. Pat. No. 7,144,703, each of which is incorporated herein by reference.
ELISA to Detect Antibody Production—Hepatitis B Surface Antigen
The antigenic effect of the hepatitis B surface antigen is determined in Balb/c mice using 5, two-fold dilutions each of vaccine subject to stability and/or stress testing (i.e., test vaccine) and vaccine not subject to stability and/or stress testing (i.e., reference vaccine). A 1.0 ml dose of test or reference vaccine is administered intraperitoneally to 10 mice per dilution. Ten mice are inoculated with diluent to serve as a placebo. Twenty-eight days after inoculation, blood is collected from the mice and the sera separated. The serum samples are tested for antibody titer against hepatitis B surface antigen using ELISA. The test vaccine passes the hepatitis B potency test if the upper limit of its relative potency is ≧1.
ELISA to Detect Antibody Production—Hepatitis a Virus Antigen and Hepatitis B Surface Antigen
The antigenic effect of the hepatitis A virus antigen and hepatitis B surface antigen is determined in 4 week old SPF guinea pigs subcutaneously immunized with 200 μl of the vaccine. Vaccine comprising only the hepatitis A viral antigen, only the hepatitis B surface antigen or a combination of both may be analyzed by this method. Ten animals are immunized with the vaccine subject to stability and/or stress testing (i.e., test vaccine) and 10 animals are immunized with vaccine not subject to stability and/or stress testing (i.e., reference vaccine). Six weeks after immunization blood is collected and the sera separated. The serum samples are tested for antibody titers against hepatitis A virus antigen and/or hepatitis B surface antigen using ELISA.
The anti-hepatitis A viral antibody titer is determined using a competitive inhibitory ELISA technique. Specifically, a microplate well is coated with anti-hepatitis A virus rabbit serum and blocked with BSA then reacted with hepatitis A viral antigen at 4° C. overnight. Thereafter, serum obtained from test or reference vaccine immunized animals is added to the well and incubated at room temperature for 30 minutes. Following this, a peroxidase-labeled anti-hepatitis A virus rabbit antibody is added to the well and incubated at 37° C. for 2 hours. The substrate solution is added for color development. The reaction is stopped and the absorbance at 492 nm is measured and the antigen titer calculated as a titer at which the inhibition rate is 50% based on the calibration curve of a standard material. The antibody used as a standard material is prepared so that it demonstrates a relative titer of 2 IU/ml when the anti-hepatitis A virus antibody titer of the Anti-Hepatitis A Reference Globin No. 1 from the Bureau of Biologics of the F.D.A is set at 100 IU/ml.
The anti-hepatitis B surface antigen antibody titer is determined using an AUSAB kit (Abbott Co., Ltd) and based on a calibration curved developed using the WHO International Reference (I-HGIB, 50 IU/ml).
Example 13: Confirmation of Stability and Activity of EPERZAN® (Albiglutide)
Albiglutide (EPERZAN®) is a glucagon-like peptide-1 (GLP-1) receptor agonist. GLP-1 is secreted by the gastrointestinal tract during eating and increases insulin secretion, decreases glycemic excursion, delays gastric emptying and reduces food intake. Albiglutide is presently indicated for treatment of adults with type 2 diabetes.
Albiglutide is a GLP-1 receptor agonist comprising a dipeptidyl peptidase-4 resistant GLP-1 dimer fused in series to human albumin Native GLP-1 peptide is rapidly degraded while albiglutide has a longer duration of action (e.g. half-life of 4 to 7 days) due to its resistance to hydrolysis by dipeptidyl peptidase-4. Albiglutide is designed for weekly or biweekly subcutaneous dosing. Albiglutide may be provided at a concentration of 50 mg/mL following resuspension of a lyophilized form comprising 2.8% mannitol, 4.2% trehalose dihydrate, 0.01% polysorbate 80, 10 to 20 mM phosphate buffer at pH 7.2. The formulation is diluted with water for injection as necessary for respective dosing.
Albiglutide samples are incubated in containers as described in the stability testing and/or stress testing methods provided in Example 6. Preferably, the samples are incubated both in standard glass vials with appropriate closures and glass vials such as those provided herein. At the desired intervals, samples are removed and assayed to determine the activity of the agent in, at least, on in vitro or in vivo assay to assess the biological activity of albiglutide. The activity of albiglutide is determined using methods and controls appropriate to the agent, for example using methods provided in Baggio et al., Diabetes 53, 2492-2500 (2004); US Publication 20110301080 A1; or any one of U.S. Pat. Nos. 7,141,547; 7,238,667; 7,592,010; 7,799,759; 7,847,079; 8,012,464; 8,071,539; 8,211,439; and 8,252,739, each of which is incorporated herein by reference.
Measurement of cAMP to Detect Activation of GLP-1 Receptor
The ability of albiglutide subject to stability and/or stress testing (i.e., test compound) or albiglutide not subject to stability and/or stress testing (i.e., reference compound) to activate the GLP-1 receptor in vitro is determined by measuring the accumulation of cAMP in baby hamster kidney cells stably transfected with rat GLP-1 receptor (BHK-GLP-1R). BHK-GLP-1R cells are grown to 70-80% confluence in 24-well plates in the absence of G418 at 37° C. Cells are incubated with Dulbecco's modified Eagle's medium containing serum and 100 μmol/L 3-isobutyl-1-methylxanthine for 5 minutes at 37° C., followed by an additional 10 minute incubation in the presence of increasing concentrations of test or reference compound. All reactions are carried out in triplicate and terminated by addition of ice-cold absolute ethanol. Cell extracts are lyophilized and cAMP levels measured using a cAMP radioimmunoassay kit (Biomedical Technologies, Stoughton Mass.).
In Vivo Activation of GLP-1 Receptor
To determine whether albiglutide subject to stability and/or stress testing or albiglutide not subject to stability and/or stress testing is capable of reaching body sites and activating GLP-1 receptor dependent processes in vivo, glucose tolerance tests, measurement of plasma insulin, feeding studies, gastric emptying studies, and c-FOS activation studies are conducted.
Glucose Tolerance Tests and Measurement of Plasma Insulin
The ability of albiglutide subject to stability and/or stress testing (i.e., test compound) or albiglutide not subject to stability and/or stress testing (i.e., reference compound) to modulate blood glucose and plasma insulin levels is measured in C57BL/6 mice. Following an overnight (16-18 hour) fast, mice are treated with test or reference compound followed by administration of 1.5 mg/g body weight of glucose orally or by injection into the peritoneal cavity. A blood sample is collected from the tail vein at 0, 10, 20, 30, 60, 90 and 120 minutes after glucose administration. Blood glucose concentrations are measured using a Glucometer Elite blood glucose meter (Bayer, Toronto, Ontario, Canada). Plasma insulin is measured in a 100 μl blood sample collected from the tail vein during the 10 and 20 minute time periods after glucose administration. The blood sample is immediately mixed with 10% volume of a chilled solution containing 5,000 KIU/ml Trasylol (Bayer), 32 mmol/l EDTA, and 0.1 nmol/l diprotin A. Plasma insulin is measured using a rat insulin enzyme-linked immunosorbent assay kit (Crystal Chem, Chicago Ill.) with mouse insulin as a standard.
Feeding Studies
The ability of albiglutide subject to stability and/or stress testing (i.e., test compound) or albiglutide not subject to stability and/or stress testing (i.e., reference compound) to inhibit food intake is measured in C57BL/6 mice. After an overnight fast (16 hours), mice are injected intracerebroventrically or intraperitoneally with control (PBS or human serum albumin), or test or reference compound. Mice receiving a 5 μl injection into the lateral ventricle are allowed to recover from anesthesia, then assessed for food intake. Mice that receive a 100 μl intraperitoneal injection are weighed and then placed into individual cages containing preweighed rodent food and water. At 2, 4, 7, and 24 hours after reagent administration, the food is reweighed and total food intake calculated.
Gastric Emptying Studies
The ability of albiglutide subject to stability and/or stress testing (i.e., test compound) or albiglutide not subject to stability and/or stress testing (i.e., reference compound) to inhibit gastric emptying is measured in C57BL/6 mice. After an overnight fast (18 hours), mice are allowed free access to preweighed food for 1 hour. After the 1 hour re-feeding, the remaining food is weighed and food intake is determined. Mice are injected intraperitoneally with PBS, human serum albumin (2.7 mg/kg), or test or reference compound (3 mg/kg) then deprived of food for an additional 4 hours. Following this, the mice are anesthetized and their stomachs removed. The stomach content wet weight is determined Gastric emptying rate is calculated using the following: gastric emptying rate (%)=[1−(stomach content wet weight/food intake)]×100.
C-Fos Activation Studies
The ability of albiglutide subject to stability and/or stress testing (i.e., test compound) or astuprotimut-R not subject to stability and/or stress testing (i.e., reference compound) to activate c-FOS expression is measured in C57BL/6 mice brains. Mice are given intraperitoneal injections of PBS, human serum albumin, albiglutide subject to stability and/or stress testing, or albiglutide not subject to stability and/or stress testing in a 100 μl volume. At 10 and 60 minutes after injection, mice are anesthetized and perfused intracardially with ice-cold normal saline followed by 4% paraformaldehyde solution. Brains are removed immediately at the end of the perfusion, kept cold in ice cold 4% paraformaldehyde solution for 3 days then transferred to a solution containing paraformaldehyde and 10% sucrose for 12 hours. Brains are cut into 25 μm sections using a Lecia SM2000R sliding microtome and stored at −20° C. in a cold cryoprotecting solution. Sections are processed for immunocytochemical detection of c-FOS using an avidin-biotin-immunoperoxidase method. The c-FOS antibody is used at a 1:50,000 dilution. Brain sections corresponding to the level of the area postrema, the nucleus of the solitary tract, the central nucleus of the amygdala and the parabrachial and paraventricular nuclei are selected for quantitative assessment of c-FOS immunoreactive neurons.
Example 14: Confirmation of Stability and Activity of MAGE-A3 Antigen-Specific Cancer Immunnotherapeutic (Astuprotimut-R)
Astuprotimut-R (MAGE-A3 Antigen-Specific Cancer Immunotherapeutic) is a form of fusion protein consisting of MAGE-A3 (melanoma associated antigen 3) epitope. MAGE-A3 may also be referred to as MAGE-3. A MAGE-A3 fusion protein may induce a therapeutic effect by stimulating a cytotoxic T lymphocyte (CTL) response against tumor cells that express the MAGE-A3 antigen, resulting in tumor cell death. Astuprotimut-R is a fusion protein consisting of the MAGE-A3 (melanoma associated antigen 3) epitope fused to part of the Haemophilus influenza protein D antigen sequence. MAGE-A3 is a tumor specific antigen expressed in a variety of cancers (e.g., melanoma, non-small cell lung cancer, head and neck cancer and bladder cancer). Astuprotimut-R is administered along with an adjuvant system comprising specific combinations of immunostimulating compounds selected to increase the anti-tumor response.
Astuprotimut-R samples are incubated in containers as described in the stability testing and/or stress testing methods provided in Example 6. Preferably, the samples are incubated both in standard glass vials with appropriate closures and glass vials such as those provided herein. At the desired intervals, samples are removed and assayed to determine the activity of the agent in, at least, on in vitro or in vivo assay to assess the biological activity of albiglutide. The activity of astuprotimut-R is determined using methods and controls appropriate to the agent, for example using methods provided in Vantomme et al., J Immunother 27, 124-135 (2004); WO99/40188; U.S. Pat. No. 7,049,413; U.S. Pat. No. 7,371,845; U.S. Pat. No. 7,388,072; and US Publication 20090203124, each of which is incorporated herein by reference.
ELISA Assay to Detect Antibody Production
The antigenic effect of astuprotimut-R is determined in 2 different mouse strains (C57BL/6 and Balb/C). Five mice of each strain are injected twice at 2 week intervals in the foot pad with 5 μg of astuprotimut-R subject to stability and/or stress testing (i.e., test vaccine) or astuprotimut-R not subject to stability and/or stress testing (i.e., reference vaccine) at 1/10th of the concentration used in human settings.
Sera are prepared from blood collected from the mice 2 weeks after the last injection. Two μg/ml of purified antigen is used as coated antigen. After saturation at 1 hour at 37° C., in PBS with 1% newborn calf serum, the sera are serially diluted (starting at 1/1000) and incubated overnight at 4° C., or 90 minutes at 37° C. After washing in PBS/Tween 20, biotinylated goat anti-mouse total IgG (1/1000) or goat anti-mouse IgG1, IgG2a, IgG2b antisera (1/5000) are used as second antibodies. After 90 minutes incubation at 37° C., streptavidin coupled to peroxidase is added, and TMB (tetra-methyl-benzidine peroxide) is used as substrate. After 10 minutes the reaction is blocked by addition of H2SO4 0.5M, and the O.D. is determined.
Proliferation Assay to Detect Immune Response
The immunogenic effect of astuprotimut-R is determined in 2 different mouse strains (C57BL/6 and Balb/C). Five mice of each strain are injected twice at 2 week intervals in the foot pad with 5 μg of astuprotimut-R subject to stability and/or stress testing (i.e., test vaccine) or astuprotimut-R not subject to stability and/or stress testing (i.e., reference vaccine) at 1/10th of the concentration used in human settings.
Lymphocytes for proliferation assays are prepared by sacrificing the mice followed by collection and crushing of the spleen or popliteal lymph nodes 2 weeks after the last injection. 2×105 cells are placed in triplicate in 96 well plates and the cells are re-stimulated in vitro for 72 hours with different concentrations (1-0.1 μg/ml) of antigen or antigen coated onto latex micro-beads. The lymphoproliferative response of mice receiving either test or reference vaccine is documented.
Gel Electrophoresis Methods for Size Detection
Samples of astuprotimut-R subject to stability and/or stress testing or not subject to stability and/or stress testing are diluted and resolved by gel electrophoresis. SDS-PAGE is used to determine protein size and aggregate formation. Native gel electrophoresis is used to analyze protein complexes formed by covalent or non-covalent interaction. Non-reducing gel electrophoresis is used to analyze protein complexes or protein structures containing disulfide bonds. Proteins resolved by gel electrophoresis are detected using sensitive staining methods, such as silver stain. Gel electrophoresis and silver staining methods are known in the art.
Example 15: Confirmation of Stability and Activity of GSK2402968 (Drisapersen)
GSK2402968 (drisapersen) is a dystrophin antisense oligonucleotide, or an analog thereof. Antisense oligonucleotides may affect the post-transcriptional splicing process of premRNA to restore the reading frame of the mRNA, resulting in a shortened dystrophin protein. Drisapersen is currently indicated for the treatment of boys with Duchenne Muscular Dystrophy with a dystrophin gene mutation amenable to an exon 51 skip. Drisapersen is an antisense oligonucleotide (5′-UCAAGGAAGAUGGCAUUUCA-3′) (SEQ ID NO: 1) with full length 2′-O-methyl-substituted ribose moieties and phosphorothioate internucleotide linkages. Binding of the antisense oligonucleotide to either one or both splice sites or exon-internal sequences induces skipping of the exon by blocking recognition of the splice sites by the spliceosome complex. Drisapersen may be provided in 0.5 ml glass vials in sodium phosphate buffered saline at a concentration of 100 mg/ml. It may be formulated for abdominal subcutaneous administration at a dose of 0.5 to 10 mg per kg of body weight.
To determine the stability of drisapersen, drisapersen samples are incubated in containers as described in the stability testing and/or stress testing methods provided in Example 6. Preferably, the samples are incubated both in standard glass vials with appropriate closures and glass vials such as those provided herein. At the desired intervals, samples are removed and assayed to determine the stability and/or activity of the agent. The stability and/or activity of drisapersen is determined using methods and controls appropriate to the agent, for example, using methods provided in Aartsma-Rus et al., Hum Molec Genet. 12, 907-914 (2003); van Deutekom et al., N Engl J Med 357, 2677-2686; Goemans et al., N Engl J Med 364, 1513-1522 (2011); US Patent Publications US 20080209581, US20090228998, US20110294753 and US20130035367; U.S. Pat. No. 7,973,015; and U.S. Pat. No. 8,304,398, each of which is incorporated herein by reference.
Gel Mobility Shift Assays to Detect Binding Affinity
The binding affinity of drisapersen subject to stability and/or stress testing (i.e., test oligo) or drisapersen not subject to stability and/or stress testing (i.e., reference oligo) for the target sequence is demonstrated using a gel mobility shift assay. The exon 51 target RNA fragment is generated by in vitro T7-transcription from a PCR fragment (amplified from either murine or human muscle mRNA using a sense primer that contains the T7 promoter sequence) in the presence of 32P-CTP. The binding affinity of test or reference oligo (0.5 pmol) for the target transcript fragments is determined by hybridization at 37° C. for 30 minutes and subsequent polyacrylamide (8%) gel electrophoresis. Drisapersen generates a mobility shift, demonstrating its binding affinity for the target RNA.
RT-PCR and Sequence Analysis to Detect Exon Skipping in Muscle Cell Cultures
The ability of drisapersen subject to stability and/or stress testing (i.e., test oligo) or drisapersen not subject to stability and/or stress testing (i.e., reference oligo) to induce skipping in muscle cells in vitro is measured in proliferating mouse myoblasts and post-mitotic mouse myotube cultures (expressing higher levels of dystrophin) derived from the mouse muscle cell line C2C12. The analysis is also conducted in human-derived primary muscle cell cultures isolated from muscle biopsies from Duchenne Muscular Dystrophy patients carrying a dystrophin gene mutation amenable to an exon 51 skip. These heterogeneous cultures contain approximately 20-40% myogenic cells. The test or reference oligo (at a concentration of 1 μM) is transfected into the cells using the cationic polymer PEI (MBI Fermentas) at a ratio-equivalent of 3. The test or reference oligo used in these experiments contains a 5′ fluorescein group which allows for the determination of the transfection efficiencies by counting the number of fluorescent nuclei. Typically, more than 60% of cells show specific nuclear uptake of the drisapersen.
Following transfection, RNA is isolated 24 hours later using RNAzol B (CamPro Scientific, The Netherlands). RNA is reverse transcribed using C. therm polymerase (Roche) and either a mouse or a human dystrophin specific reverse primer. To facilitate the detection of skipping of dystrophin exon 51, the cDNA is amplified by two rounds of PCR, including a nested amplification using either mouse or human dystrophin specific primers. A truncated product corresponding to the RNA caused by exon 51 skipping is detected. Subsequent sequence analysis confirms specific skipping of exons in the dystrophin transcripts.
Immunohistochemical Analysis to Detect Dystrophin in Human Muscle Cell Cultures
The ability of drisapersen subject to stability and/or stress testing (i.e., test oligo) or drisapersen not subject to stability and/or stress testing (i.e., reference oligo) to induce the skipping of exon 51 in muscles cells from patients carrying deletions amenable to an exon 51 skip and to restore the translation and synthesis of a dystrophin protein is shown using immunohistochemical analysis. Patient derived muscle cell culture are transfected with test or reference oligo then subjected to immunocytochemistry using two different dystrophin monoclonal antibodies raised against domains of the dystrophin protein located proximal and distal of the targeted region respectively. Fluorescent analysis reveals restoration of dystrophin synthesis in patient-derived cell cultures. Approximately at least 80% of the fibers will stain positive for dystrophin in the treated samples.
Example 16: Confirmation of Stability and Activity of HZ/Su (Herpes Zoster Virus Vaccine)
Herpes zoster virus vaccine (HZ/su) is a form of recombinant varicella zoster virus glycoprotein E. The varicella zoster virus glycoprotein E subunit is the most abundant glycoprotein in the varicella zoster virus and induces a potent CD4+ T-cell response when administered with an adjuvant. Herpes zoster virus vaccine is formulated for intramuscular injection and comprises 50 μg of varicella zoster virus glycoprotein E in 0.2 ml mixed with 0.5 ml of AS01B adjuvant. AS01B adjuvant is a liposome based adjuvant system containing 50 μg 3-O-desacyl-4′-monophosphoryl lipid A (MPL) and 50 μg QS21 (a triterpene glycoside).
To determine the stability of herpes zoster virus vaccine, herpes zoster virus vaccine samples are incubated in containers as described in the stability testing and/or stress testing methods provided in Example 6. Preferably, the samples are incubated both in standard glass vials with appropriate closures and glass vials such as those provided herein. At the desired intervals, samples are removed and assayed to determine the stability and/or activity of the agent. The stability and/or activity of herpes zoster virus vaccine is determined using methods and controls appropriate to the agent, for example, using methods provided in any of Jacquet, et al., Vaccine 20, 1593-1602 (2002); Leroux-Roels, et al., J Infect Dis 15, 1280-1290 (2012); Haumont, et al., J Med Virol 53, 63-68 (1997); and WO2006/094756, each of which is incorporated herein by reference.
ELISA to Detect Antibody Production
The antigenic effect of HZ/su is determined in guinea pigs and mice. Three groups of 6 female Durkin Hartlay guinea pigs are subcutaneously immunized twice at 28 day intervals with 8 μg of HZ/su subject to stability and/or stress testing (i.e., test vaccine), 8 μg of HZ/su not subject to stability and/or stress testing (i.e., reference vaccine), or adjuvant alone. Blood samples are collected on days 28, 42 and 72. Three groups of 6 female BalbC mice are subcutaneously immunized twice at 28 day intervals with 5 n of HZ/su subject to stability and/or stress testing (i.e., test vaccine), 5 μg of HZ/su not subject to stability and/or stress testing (i.e., reference vaccine), or adjuvant alone. Blood samples are collected on days 28 and 42. Mice are sacrificed on day 42 and their spleens collected.
Sera prepared from the blood samples is analyzed for anti-recgE antibodies by ELISA. Immunoplates are coated with recgE at 4° C. Plates are washed 5 times with TBS-Tween (50 mM Tris-HCl pH 7.5, 150 mM NaCl, 0.1% Tween 80) and saturated for 1 hour at 37° C. with 150 μl of the same buffer supplemented with 1% BSA. Serial dilutions of sera in saturation buffer are incubated for 1 hour at 37° C. Plates are washed 5 times with TBS-Tween buffer and antigen-bound antibodies are detected with the second antibody (goat anti-mouse or goat anti-guinea pig IgG) coupled to alkaline phosphatase (dilution 1/7500 in TBS-Tween buffer). The enzymatic activity is measured using the p-nitrophenylphosphate substrate dissolved in diethanolamine buffer (pH 9.8). OD 415 nm is measured in an ELISA reader.
Mouse antibody subclass is determined using immunoplates coated as described above and IgG1- or IgG2a-specific biotin-labeled monoclonal antibodies (rat anti-mouse, dilution 1/7000 in TBS-Tween buffer and 1% BSA) as second antibodies. Phosphatase alkaline-conjugated streptavidin (1/1000 dilution) is added to each well. The enzymatic activity is measured using the p-nitrophenylphosphate substrate dissolved in diethanolamine buffer (pH 9.8). OD 415 nm is measured in an ELISA reader.
ELISA titers are identified as the reciprocal of the dilution giving a signal corresponding to 50% of the maximal OD 415 nm value.
Neutralization Assay to Detect Antibody Mediated Neutralization of Varicella Zoster Virus Infectivity
The ability of HZ/su to stimulate the production of neutralizing antibodies is analyzed using sera collected from mice treated as described above. Varicella zoster virus (100 μl at 4×102 pfu/ml) is incubated for 1 hour at 37° C. with 100 μl of heat-inactivated mouse serum serially diluted in buffer (PBS, saccharose 5%, glutamate 1%, fetal bovine serum 10%, pH 7.1). Guinea pig serum complement (2 μl) is added to each well and incubated for 2 hours at 37° C. then plated onto a confluent MRC-5 monolayer at room temperature in the dark. Two hours later, 800 μl of culture medium (RPMI plus 2% fetal bovine serum) is added. Seven days later, the medium is removed, and cells are fixed and stained with Coomassie blue solution for 10 minutes. Plates are washed with distilled water and lysis plaques are counted. Each dilution is tested in duplicate. The neutralizing titer is expressed as the reciprocal of the serum dilution resulting in 50% neutralization.
It will be apparent to those skilled in the art that various modifications and variations can be made to the embodiments described herein without departing from the spirit and scope of the claimed subject matter. Thus it is intended that the specification cover the modifications and variations of the various embodiments described herein provided such modification and variations come within the scope of the appended claims and their equivalents.

## Claims

1. A packaged pharmaceutical composition comprising:
A glass pharmaceutical container and a pharmaceutical composition contained within the glass pharmaceutical container, wherein the pharmaceutical composition comprises:
Diphtheria and Tetanus Toxoids and Acellular Pertussis Adsorbed, Hepatitis B (Recombinant) and Inactivated Poliovirus Vaccine, Hepatitis A Vaccine, Hepatitis B Vaccine (Recombinant), Hepatitis A & Hepatitis B (Recombinant) Vaccine, albiglutide, MAGE-A3 Antigen-Specific Cancer Immunotherapeutic (astuprotimut-R), GSK2402968 (drisapersen), or HZ/su (herpes zoster vaccine) and a pharmaceutically acceptable excipient; wherein the glass pharmaceutical container comprises a glass composition comprising
SiO2 in a an amount greater than or equal to 72 mol. % and less than or equal to 78 mol. %;
alkaline earth oxide comprising both MgO and CaO, wherein CaO is present in an amount up to about 1.0 mol. %, and a ratio (CaO (mol. %)/(CaO (mol. %)+MgO (mol. %))) is less than or equal to 0.5;
X mol. % Al2O3, wherein X is greater than or equal to 5 mol. % and less than or equal to 7 mol. %;
Y mol. % alkali oxide, wherein the alkali oxide comprises Na2O in an amount greater than 8 mol. %; and
a ratio of a concentration of B2O3 (mol. %) in the glass container to (Y mol. %−X mol. %) is less than or equal to 0.3.

2. The packaged pharmaceutical composition of claim 1, wherein the glass pharmaceutical container comprises a compressive stress greater than or equal to 150 MPa.

3. The packaged pharmaceutical composition of claim 1, wherein the glass pharmaceutical container comprises a compressive stress greater than or equal to 250 MPa.

4. The packaged pharmaceutical composition of claim 1, wherein the glass pharmaceutical container comprises a depth of layer greater than 30 μm.

5. The packaged pharmaceutical composition of claim 1, wherein the glass pharmaceutical container maintains the stability, product integrity, or efficacy of the pharmaceutical composition.

6. The packaged pharmaceutical composition of claim 1:
wherein the glass pharmaceutical container has a compressive stress greater than or equal to 150 MPa and a depth of layer greater than 10 μm, and wherein the glass pharmaceutical container maintains the stability, product integrity, or efficacy of the pharmaceutical composition.

7. The packaged pharmaceutical composition of claim 1:
wherein the glass pharmaceutical container is substantially free of boron, and wherein the glass pharmaceutical container maintains the stability, product integrity, or efficacy of the pharmaceutical composition.

8. The packaged pharmaceutical composition of claim 7, wherein the glass pharmaceutical container comprises a compressive stress greater than or equal to 150 MPa and a depth of layer greater than 25 μm.

9. The packaged pharmaceutical composition of claim 8, wherein the glass pharmaceutical container comprises a compressive stress greater than or equal to 300 MPa and a depth of layer greater than 35 μm.

10. The packaged pharmaceutical composition of claim 7, wherein said glass pharmaceutical container comprises an internal homogeneous layer.

11. The packaged pharmaceutical composition of claim 10, wherein said glass pharmaceutical container comprises a compressive stress greater than or equal to 150 MPa and a depth of layer greater than 25 μm.

12. The packaged pharmaceutical composition of claim 1, wherein the glass pharmaceutical container comprises an internal homogeneous layer.