Patent Application
20250164424
The invention relates to a method for evaluating suitability of a pharmaceutical container for lipid-based carrier systems. Lipid-based carrier systems are sensitive pharmaceutical vehicles which require a pharmaceutical container that meets the desired standards for storage and shipment.
Lipid-based carrier systems, such as e.g. lipid nanoparticles (LNPs) are a modern drug delivery vehicle that is used for pharmaceutically active, sensitive ingredients, for example mRNA.
LNPs used for mRNA vaccines against SarS-COV-2 are based on four chemically different types of lipids, i.e. phospholipids, cholesterol, PEG-modified lipids and cationic lipids. Cationic lipids bind mRNA due to their opposite molecular charges. mRNA vaccines are chemically sensitive and require high demands on their storage conditions, for example temperatures well below −20° C. to preserve the drug.
Accordingly, high demands are placed on the containers for the storage and transport of mRNA vaccines. One particular problem in the context of mRNA vaccines is that adhesion and possible inactivation of the supramolecular structures to the container wall has been observed. In that context, there remains a challenge to assess pharmaceutical containers for their suitability for lipid-based carrier systems with respect to storage and shipment.
A wealth of pharmaceutical containers is on the market, most of which are glass-based, while some are polymer-based. Pharmaceutical containers may also be coated to alter their surface properties, in particular with respect to adhesion properties.
Yet, a method for evaluating the suitability of a pharmaceutical container for lipid-based carrier systems remains elusive.
In a first aspect, the invention relates to a method for evaluating suitability of a pharmaceutical container for lipid-based carrier systems comprising the following steps:
The method according to the invention advantageously allows to collect relevant information in the form of ToF-SIMS data from the inner surface of the container which has previously undergone a typical treatment, i.e. incubation the container with a lipid-based carrier system. The ToF-SIMS data provide a wealth of information on relevant molecular (ion) species that are found in the coating and/or on the container. The method advantageously allows to reduce this complex set of information via MCR (Multivariate Curve Resolution) analysis on the ToF-SIMS data using one or more factors that are representative of relevant compound classes found in the coating and/or on the container. The method provides a single score value for each chosen factor that can be easily compared to reference conditions.
The evaluation of the score of one or more factors by comparison to a reference pharmaceutical container or reference conditions makes this method versatile, i.e. an experimental frame of reference may be chosen which suits the pharmaceutical container, e.g. a coated container may be compared over an uncoated container. Further, the method allows free choice of mathematical evaluation, e.g. by assessing difference or quotient values. Also, threshold values may differ for certain applications and requirements set by the authorities.
In a related aspect, the invention relates to a method for evaluating suitability of a pharmaceutical container for lipid-based carrier systems comprising the following steps:
In a related aspect, the invention relates to a method for evaluating suitability of a pharmaceutical container for lipid-based carrier systems comprising the following steps:
In a related aspect, the invention relates to a method for evaluating suitability of a pharmaceutical container for lipid-based carrier systems comprising the following steps:
In a first aspect, the invention relates to a method for evaluating suitability of a pharmaceutical container for lipid-based carrier systems comprising the following steps:
The acquired ToF-SIMS data uncover a multitude of molecular (ion) species signals which originate from the container and/or the coating, e.g. the glass or a polymer substrate, siloxanes, phospolipids, other lipids, buffer ingredients, as well as unspecific organic molecules and inorganic salts. The high-dimensional data space created by the secondary ions, which are treated as individual variables, are then replaced by new variables, the so-called factors. The position of the factors within the high-dimensional data space is described by loadings. The application of MCR (Multivariate Curve Resolution) analysis on the loadings with respect to one or more of the (specific) factors yields a score.
The skilled person knows that ToF-SIMS provides masses of molecular fragments which are detected by the instrument. Subsequently, using analysis tools and/or appropriate libraries, the obtained masses are assigned to, or interpreted as, specific molecular (ion) species.
In one embodiment, the method comprises selecting between 10 to 1000 signals, or between 20 to 800 signals, or between 50 to 500 signals, or between 100 to 200 signals, from the ToF-SIMS data. The skilled person knows how to choose relevant molecular (ion) species from ToF-SIMS data depending on the investigated container. In one embodiment, the method comprises selecting 10 or more signals, or 20 or more signals, or 50 or more signals, or 100 or more signals. In one embodiment, the method comprises selecting 1000 signals or less, 800 signals or less, 500 signals or less, or 200 signals or less.
In one embodiment, the method comprises selecting one or more factors, e.g. 3 to 8 factors, three factors, four factors, five factors, or three to five factors, wherein the factors are selected from lipid factor1, silicon-organic factor1, silicon-inorganic factor1 and organic factor1.
In one embodiment, the pharmaceutical container is a syringe, a cartridge, an ampoule or a glass vial, wherein the pharmaceutical container is a glass container or a polymer container.
In one embodiment, the step incubating the container with a lipid-based carrier system comprises the following steps:
In one embodiment of the method, the filled container is stored for a period of at least 3 hours, or at least 6 hours;
In one embodiment of the method, before filling the container with the reference composition, the container is cleaned, preferably with water.
In one embodiment of the method, the reference composition
In one embodiment, the step incubating the container with a lipid-based carrier system comprises the following steps:
The incubation step with a lipid-based carrier system may differ for a glass container vis-à-vis a polymer container and is performed under well-defined conditions to establish reproducibility and comparison between the container and a reference container.
In one embodiment, the reference LNP-composition is the Comirnaty vaccine (license number EU/1/20/1528), or
In one embodiment, the reference LNP-composition contains 7.2 mg/ml (4-hydroxybutyl)azanediyl)bis(hexane-6,1-diyl)bis(2-hexyldecanoate), 0.83 mg/mL 2 [(polyethylene glycol)-2000]-N, N-ditetradecylacetamide, 1.5 mg/mL 1,2-distearoyl-sn-glycero-3-phosphocholine, and 3.3 mg/mL cholesterol, in phosphate-buffered saline (PBS) (pH 7.4) with a saccharide content of 10 wt. % and the following concentrations:
In one embodiment, the one or more factors further include silicon-organic factor1, silicon-inorganic factor1 and organic factor1.
In one embodiment, the step performing MCR analysis on the ToF-SIMS data, wherein each factor has a factor-specific MCR loading which indicates a conceptional component in an n-dimensional compositional space which can be attributed to each of the one or more factors, wherein the factor-specific MCR loading characterizes the one or more factors by listing the ions/masses that contribute to the definition of said factor.
The one or more factors may be selected from lipid factor1, silicon-organic factor1, silicon-inorganic factor1 and organic factor1, wherein each of the factors has a factor-specific MCR loading which indicates a conceptional component in said n-dimensional compositional space which can be attributed to said one or more factors, wherein the factor-specific MCR loading characterizes the one or more factors by listing the ions that contribute to the definition of said factor. The conceptional component represents compound classes, such as e.g. lipids, siloxanes, and glass-typical silicon species. As such the conceptional component is not present in the coating or on the container.
In one embodiment, the lipid factor1 includes, in its factor specific MCR loading, one or more of the following ions: fatty acid-ions; [CnH2n-1O2]−, wherein n is 10, 12, 14, 16 or 18; [CnH2n-3O2]−, wherein n is 10, 12, 14, 16 or 18; [CnH2n-5O2]−, wherein n is 16 or 18; phosphatidyl-choline ions; [(CH)nH2O4P]−, wherein n=0, 1, 2 or 3.
In one embodiment, the silicon-organic factor1 includes, in its factor specific MCR loading, one or more of the following ions: silane species, silicon-carbon species, polysiloxane species based on the formula [OSiR1R2]n−, wherein R1 and R2 are independently selected from methyl, ethyl, propyl, and wherein n is any integer between 2 and 10.
In one embodiment, the silicon-inorganic factor1 includes, in its factor specific MCR loading, one or more of the following ions: silicon species; aluminium-species and/or boron species; halogen species.
In one embodiment, the lipid factor1 includes, in its factor specific MCR loading, one or more of the following ions: [C10H17O2]−, [C10H19O2]−, [C12H21O2]−, [C16H29O2]−, [C16H31O2]−, [C16H32O2]−, [C18H31O2]−, [C18H33O2]−, [C18H35O2]−, [PO3]−, [PH2O4]−, [CH3O4P]−, [C2H4O4P]−.
In one embodiment, the silicon-organic factor1 includes, in its factor specific MCR loading, one or more of the following ions: [Sic]−, [SiCH3O]−, [SiCH3O2]−, [SiC2H5O]−, [Si2CHO2]−, [SiC3H9O]−, [Si2C5H15O2]−, [Si3C5H15O4]−.
In one embodiment, the silicon-inorganic factor1 includes, in its factor specific MCR loading, one or more of the following ions: OH−, Al−, Si−, P−, Cl−, NaO−, AlO−, BO2−, SiHO−, AlO2−, SiO2−, SiH5O2−; Si3H3O2−, Si2HO5−.
In one embodiment, the lipid factor1 includes, in its factor specific MCR loading, at least 5 of the following ions: [C10H19O2]−, [C12H21O2]−, [C16H29O2]−, [C16H31O2]−, and [C18H35O2]−.
In one embodiment, the silicon-organic factor1 includes, in its factor specific MCR loading, at least 5 of the following ions: [SiCH3O], [SiCH3O2]−, [SiC2H5O]−, [SiC3H9O]−, and [Si2C5H15O2]−.
In one embodiment, the silicon-inorganic factor1 includes, in its factor specific MCR loading, at least 5 of the following ions: OH−, Si−, SiO2−, SiH5O2−, and Si3H3O2−.
Method according to any one of the preceding claims, wherein the step evaluating the score of the one or more factors by comparison to a reference score comprises one or more of the following steps
The “reference container”, as used herein, may be an uncoated container. The reference container may be of the same dimensions and materials and bulk composition as the pharmaceutical container. For example, the pharmaceutical container may be coated whereas the reference container may be uncoated.
In one embodiment of the method, the step evaluating the score of the one or more factors by comparison to a reference score comprises one or more of the following steps:
In one embodiment of the method, the evaluation results in a positive or negative answer which, respectively, indicates fitness of suitability or lack of suitability.
In one embodiment of the method, the comparison to a reference pharmaceutical container comprises the following steps:
Advantageously, the method according to the invention can be adapted to select individual factors of relevance, such as lipid factor1, silicon-organic factor1, silicon-inorganic factor1 and organic factor1. The ToF-SIMS data acquired from the inner surface of the container cover a wealth of molecular (ion) species/masses, most of which belong to at least one well-defined compound group covered by the factors. Feeding the ToF-SIMS data into the MCR (Multivariate Curve Resolution) analysis on the ToF-SIMS data using a selected factor allows direct comparison of a pharmaceutical container to a reference pharmaceutical container by means of the corresponding scores.
It is advantageous to directly compare the scores for the pharmaceutical container and the reference pharmaceutical container by a calculus-based operation and subsequent assessment of the calculated result. This calculus-based operation and subsequent assessment can be done for the individual factors, but is also accessible for combinations of several individual factors.
In a related aspect, the invention relates to a method for evaluating suitability of a pharmaceutical container for lipid-based carrier systems comprising the following steps:
In a related aspect, the invention relates to a method for evaluating suitability of a pharmaceutical container for lipid-based carrier systems comprising the following steps:
In a related aspect, the invention relates to a method for evaluating suitability of a pharmaceutical container for lipid-based carrier systems comprising the following steps:
In one embodiment of the method, the suitability of the pharmaceutical container is evaluated based on
Pharmaceutical containers with a combined low quotient for lipid-factor1 and an elevated quotient for silicon-organic factor1 may be considered as very suitable for lipid-based carrier systems, because the results indicate that there is only little adsorption of the lipid-based carrier and its individual constituting components, while a previously provided coating of the pharmaceutical container remains intact. An intact coating of the pharmaceutical may be evidenced by silicon-organic species, such as siloxane, which are intended to protect the lipid-based carrier from adsorption to the glass.
In one embodiment of the method, the suitability of the pharmaceutical container is evaluated based on
Analogously, pharmaceutical containers with a combined low quotient for lipid-factor1 and an elevated quotient for organic factor1 may be considered as very suitable for lipid-based carrier systems, because the results indicate that there is only little adsorption of the lipid-based carrier and its individual constituting components, while the pharmaceutical polymer container remains intact. An intact polymer container may be evidenced by organic species which evolve from the polymer itself.
The following disclosure relates to the pharmaceutical container and/or the reference container.
In one embodiment, the container is a glass container or a polymer container.
In one embodiment, the container comprises a cyclic olefin copolymer. In one embodiment, the container comprises a cyclic olefin polymer.
In one embodiment, the container has one or more of the following properties:
In one embodiment, the container has a wall thickness of 0.50 mm or more, 1.00 mm or more, or 2.0 mm or more. In one embodiment, the container has a wall thickness of 10.0 mm or less, or 7.00 mm or less, or 4.0 mm or less.
In one embodiment, the container is a syringe, a cartridge, an ampoule or a vial.
The following disclosure relates to the pharmaceutical container and/or the reference container.
In one embodiment, the container comprises a glass composition comprising 50 to 90 wt. % SiO2, and 3 to 25 wt. % B2O3.
In one embodiment, the container comprises a glass composition comprising aluminosilicate, optionally comprising 55 to 75 wt. % SiO2, and 11.0 to 25.0 wt. % Al2O3.
In one embodiment, the container comprises a glass composition comprising 70 to 81 wt. % SiO2, 1 to 10 wt. % Al2O3, 6 to 14 wt. B2O3, 3 to 10 wt. % Na2O, 0 to 3 wt. % K2O, 0 to 1 wt. % Li2O, 0 to 3 wt. % MgO, 0 to 3 wt. % CaO, and 0 to 5 wt. % BaO.
In one embodiment, the container comprises a glass composition comprising 72 to 82 wt. % SiO2, 5 to 8 wt. % Al2O3, 3 to 6 wt. B2O3, 2 to 6 wt. % Na2O, 3 to 9 wt. % K2O, 0 to 1 wt. % Li2O, 0 to 1 wt. % MgO, and 0 to 1 wt. % CaO.
In one embodiment, the container comprises a glass composition comprising 60 to 78 wt. % SiO2, 7 to 15 wt. B2O3, 0 to 4 wt. % Na2O, 3 to 12 wt. % K2O, 0 to 2 wt. % Li2O, 0 to 2 wt. % MgO, 0 to 2 wt. % CaO, 0 to 3 wt. % BaO, and 4 to 9 wt. % ZrO2.
In one embodiment, the container comprises a glass composition comprising 50 to 70 wt. % SiO2, 10 to 26 wt. % Al2O3, 1 to 14 wt. B2O3, 0 to 15 wt. % MgO, 2 to 12 wt. % CaO, 0 to 10 wt. % BaO, 0 to 2 wt. % SrO, 0 to 8 wt. % ZnO, and 0 to 2 wt. % ZrO2.
In one embodiment, the container comprises a glass composition comprising 55 to 70 wt. % SiO2, 11 to 25 wt. % Al2O3, 0 to 10 wt. % MgO, 1 to 20 wt. % CaO, 0 to 10 wt. % BaO, 0 to 8.5 wt. % SrO, 0 to 5 wt. % ZnO, 0 to 5 wt. % ZrO2, and 0 to 5 wt. % TiO2.
In one embodiment, the container comprises a glass composition comprising 65 to 72 wt. % SiO2, 11 to 17 wt. % Al2O3, 0.1 to 8 wt. % Na2O, 0 to 8 wt. % K2O, 3 to 8 wt. % MgO, 4 to 12 wt. % CaO, and 0 to 10 wt. % ZnO.
In one embodiment, the container comprises a glass composition comprising 64 to 78 wt. % SiO2, 4 to 14 wt. % Al2O3, 0 to 4 wt. % B2O3, 6 to 14 wt. % Na2O, 0 to 3 wt. % K2O, 0 to 10 wt. % MgO, 0 to 15 wt. % CaO, 0 to 2 wt. % ZrO2, and 0 to 2 wt. % TiO2.
In one embodiment, the method comprises the steps:
In one embodiment, the method comprises the steps:
In one embodiment, the method comprises the steps:
In one embodiment, the method comprises the steps:
An embodiment of this disclosure relates to a method comprising the following step(s):
A further method of this disclosure includes the following steps:
An aspect of this disclosure relates to the use of the results of the evaluation obtainable by the method according to this disclosure to evaluate the suitability of a coated container, preferably a coated pharmaceutical container, for the storage of a pharmaceutical composition, preferably a pharmaceutical composition comprising a lipid-based carrier system, and/or the quality control of the production of a coated container.
Another aspect relates to a pharmaceutical container, having attached thereto, e.g. on a label, one or more results of the method according to this disclosure, wherein optionally the result is
This disclosure also relates to a kit, comprising:
Any reference to “LNP incubated” in this disclosure means that the container or coating was incubated with LNPs before measurement. If a score is denoted as a “relative” score ratio, the respective values are to be understood as being the relative ratio of the score value of the coated container divided by the score value of an uncoated reference container based on the same MCR factor. E.g., for measuring a relative LNP-incubated MCR score ratio between a coated container and an uncoated container, wherein the uncoated container is a reference container, both containers are incubated with the same specific LNP composition as applied for the coated container. Both the coated container and the reference container are analysed via ToF-SIMS and MCR to obtain absolute MCR scores, e.g. of lipid factor1. The relative LNP-incubated MCR score ratio, e.g. of lipid factor1, is obtained by dividing the resulting MCR score of the coated container by the MCR score of the reference container. For example, the relative LNP-incubated MCR score ratio of lipid factor1 may be less than 0.5. As mentioned before, the reference container may be an uncoated container. The reference container may be of the same dimensions and materials and bulk composition as the coated container (except for the coating of course).
In an embodiment, LNP-incubation of a glass container, either being an uncoated glass container or a coated glass container, comprises cleaning the container with UltraPure water (purity 1 analogue DIN ISO 3696 with ≤0.1 μS/cm at 25° C.), drying under laminar-flow conditions, incubating the container with a reference LNP-composition by filling the container with the reference LNP-composition, freezing to −80° C., incubating for 12 hours at −80° C., and then thawing to 5° C. within 12 hours, and then emptying the containing followed by a cleaning step of the inner container surface by rinsing 10 times with ultrapure water and subsequent drying under laminar flow.
In a related embodiment, LNP-incubation of a polymer container, either being an uncoated polymer container or a coated polymer container, comprises incubating the container with a reference LNP-composition by filling the container with the reference LNP-composition, freezing to −80° C., incubating for 12 hours at −80° C., and then thawing to 5° C. within 12 hours and then emptying the containing followed by a cleaning step of the inner container surface by rinsing 10 times with ultrapure water and subsequent drying under laminar flow.
In one embodiment, the pharmaceutical container has an absolute LNP-incubated MCR score of lipid factor1 of less than 7×1013, less than 5×1013, or less than 2×1013. Assessing suitability of the pharmaceutical container for lipid-based carrier may be based on this criterion and the indicated score values, i.e. the container may be considered suitable, if the score is within the indicated limit.
In one embodiment, the pharmaceutical container has a relative LNP-incubated MCR score ratio of lipid factor1 of less than 0.67, less than 0.5, less than 0.3 or less than 0.13. Assessing suitability of the pharmaceutical container for lipid-based carrier may be based on this criterion and the indicated score values, i.e. the container may be considered suitable, if the score is within the indicated limit. In one embodiment, the pharmaceutical container has an absolute LNP-incubated MCR score of silicon-organic factor1 of at least 1×1012, and/or a relative LNP-incubated MCR score ratio of silicon-organic factor1 of at least 2. Assessing suitability of the pharmaceutical container for lipid-based carrier may be based on this criterion and the indicated score values, i.e. the container may be considered suitable, if the score is within the indicated limit.
In one embodiment, the pharmaceutical container has an absolute LNP-incubated MCR score of silicon-inorganic factor1 of up to 1×1013, up to 5×1012, or up to 3×1012. Optionally, this score may be at least 0.5×1012. Assessing suitability of the pharmaceutical container for lipid-based carrier may be based on this criterion and the indicated score values, i.e. the container may be considered suitable, if the score is within the indicated limit.
In an embodiment, the relative LNP-incubated MCR score ratio of silicon-inorganic factor1 is up to 5, up to 3, or up to 1.5. Optionally, this score ratio is at least 0.1, or at least 0.2. Assessing suitability of the pharmaceutical container for lipid-based carrier may be based on this criterion and the indicated score values, i.e. the container may be considered suitable, if the score is within the indicated limit.
In one embodiment, the pharmaceutical container has an absolute LNP-incubated MCR score of silicon-organic factor1 of at least 1×1012, at least 2×1012, or at least 3×1012. Optionally, the absolute LNP-incubated MCR score of silicon-organic factor1 may reach up to 9×1013, up to 7×1013, or up to 6×1013. Assessing suitability of the pharmaceutical container for lipid-based carrier may be based on this criterion and the indicated score values, i.e. the container may be considered suitable, if the score is within the indicated limit.
In an embodiment, the pharmaceutical container has a relative LNP-incubated MCR score ratio of silicon-organic factor1 of at least 2, at least 3, or at least 5. Optionally, the relative LNP-incubated MCR score ratio of silicon-organic factor1 may be up to 20, up to 15, or up to 10. Assessing suitability of the pharmaceutical container for lipid-based carrier may be based on this criterion and the indicated score values, i.e. the container may be considered suitable, if the score is within the indicated limit.
In one embodiment, the pharmaceutical container has an absolute LNP-incubated MCR score of silicon-inorganic factor1 of at least 1×1013, and/or a relative LNP-incubated MCR score ratio of silicon-inorganic factor1 of up to 5. Assessing suitability of the pharmaceutical container for lipid-based carrier may be based on this criterion and the indicated score values, i.e. the container may be considered suitable, if the score is within the indicated limit.
In one embodiment, the container has an absolute LNP-incubated MCR score of organic factor1 of at least 1×1012, and/or a relative LNP-incubated MCR score ratio of organic factor1 of at least 0.2. Assessing suitability of the pharmaceutical container for lipid-based carrier may be based on this criterion and the indicated score values, i.e. the container may be considered suitable, if the score is within the indicated limit.
Optionally, the container has an absolute LNP-incubated MCR score of organic factor1 of up to 9×1012, up to 8×1012, or up to 6×1012. Assessing suitability of the pharmaceutical container for lipid-based carrier may be based on this criterion and the indicated score values, i.e. the container may be considered suitable, if the score is within the indicated limit.
Alternatively or in addition, the container may have corresponding, non-LNP-incubated score values. These values are obtained without LNP-incubation (“non-incubated”).
An absolute non-incubated MCR score of silicon-organic factor1 may be at least 3×1012, at least 5×1012, or at least 7×1012. Optionally, the absolute non-incubated MCR score of silicon-organic factor1 may reach up to 9×1013, up to 7×1013, or up to 6×1013. Assessing suitability of the pharmaceutical container for lipid-based carrier may be based on this criterion and the indicated score values, i.e. the container may be considered suitable, if the score is within the indicated limit.
A relative non-incubated MCR score ratio of silicon-organic factor1 may be at least 3×1012, at least 5×1012, or at least 7×1012. Optionally, the absolute LNP-incubated MCR score of silicon-organic factor1 may reach up to 9×1013, up to 7×1013, or up to 6×1013. Assessing suitability of the pharmaceutical container for lipid-based carrier may be based on this criterion and the indicated score values, i.e. the container may be considered suitable, if the score is within the indicated limit.
An absolute non-incubated MCR score of silicon-inorganic factor1 may be up to 1×1013, up to 5×1012, or up to 3×1012. Optionally, this score may be at least 0.5×1012. Assessing suitability of the pharmaceutical container for lipid-based carrier may be based on this criterion and the indicated score values, i.e. the container may be considered suitable, if the score is within the indicated limit.
A relative non-incubated MCR score ratio of silicon-inorganic factor1 may be up to 5, up to 3, or up to 1.5. Optionally, this score ratio is at least 0.1, or at least 0.2. Assessing suitability of the pharmaceutical container for lipid-based carrier may be based on this criterion and the indicated score values, i.e. the container may be considered suitable, if the score is within the indicated limit.
In an embodiment, assessing suitability of the pharmaceutical container for lipid-based carrier may be based on one or more, or all, of the above criteria/score values.
A coated glass container and an uncoated glass container (the latter serving as a reference container), both having the same dimensions, same glass type and glass composition, are treated under the exact same conditions.
For measuring the LNP-incubated MCR scores, the respective container is cleaned with UltraPure water (purity 1 analogue DIN ISO 3696 with ≤0.1 μS/cm at 25° C.) and dried under laminar-flow conditions. The container is then filled with a reference LNP-composition, frozen to −80° C., incubated for 12 hours at −80° C., and then thawed to 5° C. within 12 hours.
In a first variant, the reference LNP-composition is the Comirnaty vaccine (license number EU/1/20/1528).
In a second variant, the reference-LNP contains the following lipids in the indicated amounts: 7.2 mg/mL (4-hydroxybutyl)azanediyl)bis(hexane-6,1-diyl)bis(2-hexyldecanoate), 0.83 mg/mL 2 [(polyethylene glycol)-2000]-N,N-ditetradecylacetamide, 1.5 mg/mL 1,2-distearoyl-sn-glycero-3-phosphocholine, and 3.3 mg/mL cholesterol, in phosphate-buffered saline (PBS) (pH 7.4) with a saccharide content of 10 wt. % and the following concentrations.
Both containers are analysed via ToF-SIMS and MCR analysis (see next sections).
An absolute LNP-incubated MCR score means the MCR score obtained from a single container, either the coated glass container of the uncoated (reference) glass container, for one specific factor, wherein the factor is selected from lipid factor1, silicon-organic factor1, silicon-inorganic factor1 and organic factor1, each factor having a factor-specific MCR loading.
The relative LNP-incubated MCR score ratio refers to the quotient of the MCR score of a specific factor between the coated glass container and the uncoated (reference) glass container.
A coated polymer container and an uncoated polymer container, both having the same dimensions, same glass type and glass composition, are treated under the exact same conditions.
The polymer container preparation is the same as for the glass container preparation, except that the cleaning with UltraPure water (purity 1 analogue DIN ISO 3696 with ≤0.1 μS/cm at 25° C.) and drying under laminar-flow conditions is omitted.
In the following the measuring method and the data evaluation of the specific ToF-SIMS measurement is explained in detail. For the measurement a TOF.SIMS 4 from Iontof was used. If not stated otherwise, the ToF-SIMS are measured according to ASTM E 1829 und ASTM E 2695.
The following parameter settings were used for the ToF-SIMS analysis:
primary ion: Ga+; (alternatively using a TOF.SIMS 5 and Bi+);
(primary ion) energy: 25000V;
measuring area: 100×100 μm2;
primary ion dose density: 6×1012 cm−2;
surface discharge: via low-energetic electrons.
Spectral data were acquired for 100 s with subsequent integration.
Each sample of a (coated) container was cut lengthwise in two pieces and positioned in such a way that the centerline of the primary ion gun of the ToF-SIMS apparatus hit the inner surface of the sample. The thus generated intrinsically ionised secondary ions have been analysed via Time-of-flight analysis and separated into different detectable mass/charge ratios. Accordingly, a high-resolution mass spectrum (Δm/m>3000 for Si) is obtained covering both atomic and molecular ion species. Die surface sensitivity covers several few monolayers.
The ToF-SIMS data include n datasets consisting of ion-specific masses and their corresponding intensities. Ion species/masses are selected from the raw ToF-SIMS data, including their intensities, as indicated in
For the identification of MCR factors, the ToF-SIMS result are subjected to a subsequent multivariate analysis via MCR (Multivariate Curve Resolution). MCR is a statistical analysis method, which in its most general approach decomposes a two-way data matrix D (m×n) into two matrices C (m×k) and ST (k×n), containing respectively pure concentration profiles and pure spectra of the k species of an unknown mixture, according to the equation D=CST+E, wherein E is an error matrix containing the residuals of the data (Ruckebusch & Blanchet, Analytica Chimica Acta 765, 2013, 28-36). The MCR method has been adapted to decompose and analyse ToF-SIMS. Commercially available software can be used for this task, e.g. the software package SurfaceLab Ver 7.1., wherein optionally the number of factors is set to 3, 4 or 5. A general account of how spectral information can be dissected is given by Juan & Tauler (Analytica Chimica Acta 1145, 2021, 59-78).
Summing up, the ToF-SIMS result measured in a specific coating or container can be attributed a specific position in an n-dimensional compositional space. MCR is used to reduce the complexity of the ToF-SIMS result by summarizing the datasets into a more limited number of variables, the so-called “factors”. The results of the MCR are a set of factors, loadings and corresponding scores. Each of the factors has a factor-specific MCR loading, indicating a conceptional component in said n-dimensional compositional space which can be attributed to said factor, wherein the loading characterizes the factor in that it lists the ions that contribute to the definition of said factor. Each factor relates to substances present in or on the coating or container. To be clear, the conceptional component is not in fact present in the coating or container. Each score indicates the intensity of the corresponding factor. It correlates with the abundance of substances in or on the coating or container.
The coated glass vial was treated with either a phosphate-buffered saline (PBS) solution or a Reference-LNP in the same PBS solution. The differently treated coated glass vial were subjected to the above described ToF-SIMS measurement and data were extracted according to the above described MCR analysis.
In a first variant, the reference LNP-composition was the Comirnaty vaccine (license number EU/1/20/1528).
In a second variant, the reference-LNP contained the following lipids in the indicated amounts: 7.2 mg/mL (4-hydroxybutyl)azanediyl)bis(hexane-6,1-diyl)bis(2-hexyldecanoate), 0.83 mg/mL 2 [(polyethylene glycol)-2000]-N,N-ditetradecylacetamide, 1.5 mg/mL 1,2-distearoyl-sn-glycero-3-phosphocholine, and 3.3 mg/mL cholesterol, in phosphate-buffered saline (PBS) (pH 7.4) with a saccharide content of 10 wt. % and the following concentrations
LNPs with similar formulations may alternatively be used which formulations may deviate up to 30 wt. % from the given quantities of individual lipid components.
Additionally the LNPs contain RNA, such as mRNA, particularly based on polynucleotides containing adenine.
A method for evaluating suitability of a pharmaceutical container for lipid-based carrier systems. Lipid-based carrier systems are sensitive pharmaceutical vehicles which require a pharmaceutical container that meets the necessary standards for storage and shipment.
| Filing Document | Filing Date | Country | Kind |
|---|---|---|---|
| PCT/EP2022/058035 | 3/25/2022 | WO |
