System and method for characterization of lypohilized and other solid products

Abstract

The disclosure relates to a system and method for predicting the stability of an injectable product.

Description

SYSTEM AND METHOD FOR CHARACTERIZATION OF LYPOHILIZED AND OTHER SOLID PRODUCTSCROSS-REFERENCE TO RELATED APPLICATIONS

[0001] This application claims the benefit of priority from U.S. Appl. No. 63 / 677,821 , filed July 31 , 2024, which is incorporated by reference as if fully set forth herein.BACKGROUND

[0002] Lyophilized pharmaceutical products had a market value of >$40 B in 2018. Lyophilized products are in sealed vials that must remain intact to ensure sterility. This significantly limits the ability to characterize the lyophilized product in situ. Currently, the quality of the vial contents are determined by a visual inspection of the lyophilized product, which is called a “cake.” Most other characterization techniques require that the vial be opened and the contents analyzed, which both destroys the sterility and the contents of the vial. For example, Karl Fischer (KF) titration, which is the most common method for determining the amount of water in a product, is destructive, difficult to perform, unreliable, and provides only information about the total water content. New methods for determining the water content, stability, and / or quality of a lyophilized product are therefore needed.SUMMARY

[0003] The instant disclosure generally relates to methods that use of1H NMR spectroscopy to analyze intact (e.g., in an unopened vial) lyophilized products in situ, providing significant new information about the product quality beyond the visual inspection of the cake and the total water content via KF titration. Specifically, the location of the water in the product can be discerned, which can be critical to determining the likelihood that the product will remain stable overtime. For example, water that is bound in the product will show little or no signal using the technology, whereas free or less bound water will show a significant signal in the product. Moreover, an analysis of lyophilized products can be performed relatively rapidly (potentially in less than a minute) and without removing the product from its container (e.g., a vial). This means that every vial from a lyophilized batch can be measured for product quality.

[0004] Preliminary data has shown that the1H NMR spectrum of a lyophilized product provides significantly more information about the product beyond simply water content. For example, vials from different locations in the lyophilizer have been shown to have different product qualities, with vials in the center of the lyophilizer having potentially a higher free water content and potentially less stable.Data have shown that one can determine the difference between vials from the edge compared to vials in the center.DESCRIPTION OF THE DRAWINGS

[0005] The drawings illustrate generally, by way of example, but not by way of limitation, various embodiments discussed herein.

[0006] FIG. 1 are1H NMR spectra of an empty vial and water standards.

[0007] FIG. 2A is a photograph of a first generation1H NMR probe specially designed to perform the methods of the disclosure.

[0008] FIG. 2B is a photograph of a second generation1H NMR probe specially designed to perform the methods of the disclosure.

[0009] FIGS. 3-4 are1H NMR spectra taken three days apart for the three lyophilized samples shown in the photographs.

[0010] FIGS. 5 are1H NMR spectra taken three days apart for the three lyophilized samples shown in the photographs. In FIG. 5, the spectra in FIGS. 3-4 are placed side-by-side for easier comparison.

[0011] FIG. 6 is a 20 x 16 grid showing the location in a lyophilizer of 320 samples, with samples 2 and 4 located in the center-most region of the lyophilizer and sample 1 being closest to the outer edge of the lyophilizer.

[0012] FIG. 7 are1H NMR spectra for the four lyophilized samples shown in the photographs. Samples 1-4 are the same samples shown in FIG. 6.

[0013] FIGS. 8A-8D are1H free induction decays for four lyophilized samples taken using a time-domain (TD) NMR spectrometer.

[0014] FIG. 9 is a schematic of a high throughput system according to the disclosure.

[0015] FIG. 10A is a plot of human serum albumin (HSA) percent monomer versus time at 50°C.

[0016] FIG. 10B is1H NMR spectra obtained for lyophilized bovine serum albumin (BSA) and BSA / sucrose samples.

[0017] Unless otherwise indicated, all figures and drawings in this document are not to scale and are chosen for the purpose of illustrating different embodiments of the invention. The dimensions of the various components are depicted in illustrative terms only, and no relationship between the dimensions of the various components should be inferred from the drawings, unless so indicated. Although terms such as “top”, “bottom”, “upper”, “lower”, “under”, “over”, “front”, “back”, “up” and “down”, and “first” and “second” can be used in this disclosure, it should be understood that those terms are used in their relative sense only unless otherwise noted.DESCRIPTION

[0018] Reference will now be made in detail to certain embodiments of the disclosed subject matter. While the disclosed subject matter will be described in conjunction with the enumerated claims, it will be understood that the exemplified subject matter is not intended to limit the claims to the disclosed subject matter.

[0019] Currently there are few methods for the analysis of intact lyophilized pharmaceutical products, except for very limited non-destructive methods such as visual inspection of the cake and measuring the components of the headspace above a lyophilized cake. Alternative methods, such as evaluating reconstitution time, measuring pH, measuring osmolarity, measuring water content by KF titration, thermogravimetric analysis, differential scanning calorimetry, and size exclusion chromatography / activity are all destructive in that one cannot repeat the same test again on the same vial after measurement and one cannot use the sample after measurement. Further, none of these techniques provide information about where the water is in the lyophilized product, which can be critical to predicting product stability. Free water is generally detrimental to product stability, whereas bound water has little impact on stability.

[0020] NMR spectroscopy has been shown to be a powerful method for the analysis of lyophilized pharmaceutical products. For example, NMR spectroscopy has been used to determine mobility and phase separation of lyophilized products. In this approach, the1H Ti and1H Tirho values of the sample have been shown to correlate with product stability, especially when a bi-exponential relaxation rate indicates phase separation, and shorter1H Ti relaxation times correlate with poorer stability. However, these measurements have required the removal of a lyophilized sample from a sealed vial and packed into a solid-state NMR rotor for analysis.

[0021] The total water content in a lyophilized product has previously been determined by1H NMR spectroscopy. In that study, the authors showed that low field or time domain (TD) NMR correlated with the amount of water present in a sample, as determined by Karl Fischer titration. They were able to show that the minimum amount of water that could be measured was around 3%, and their data showed some correlation with water content. They also highlighted many of the advantages of using NMR spectroscopy to analyze lyophilized products, such as being fast, noninvasive, nondestructive, with minimal or no sample preparation, and that it is a bulk measurement of the cake for on-line or at-line measurements. The authors also compared the advantages and disadvantages of TD NMR ascompared to Karl Fischer analysis. While product quality was mentioned, it was only in the context of the overall water content of the sample.

[0022] In the investigation of lyophilized samples using1H NMR spectroscopy, it was found that samples containing similar amounts of water displayed dramatically different NMR signals. More specifically, the NMR signals differed in both appearance and intensity, which in certain cases were related to the lyophilized cake quality as assessed by the NMR signal intensity. In most cases the NMR signal was substantially weaker than the signal assessed for the total water content, indicating that the signal corresponded to only the mobile component of the sample, likely the free or unbound water. In certain cases the amount of free water approached the signal intensity expected for free water in the lyophilized sample, but mostly for cases where there were noticeable differences in the cake appearance due to poor product quality. In one example, there was a slight difference in cake appearance that was correlated with a dramatic change in the NMR spectrum.

[0023] The described method provides information about whether water in a lyophilized product is free or bound, and can do so non-destructively, which has advantages in that each lyophilized vial can be measured for product quality. It can also be used in product development, as products can be analyzed both prior to being put on stability and after being stored to monitor the changes in the product over time. It also allows other metrics, such as1H relaxation times, to be determined, which will provide additional information about sample mobility, composition, and stability.

[0024] The described method can also be implemented on a system designed for the rapid analysis of samples. Specifically, a high-throughput system is envisioned incorporating a rapid analysis of samples such that the samples are analyzed in an NMR spectrometer either at low field (less than or equal to 80 MHz1H NMR frequency) or at high field (greater than 80 MHz1H NMR frequency). Figure 9 shows one potential embodiment of a system designed for a high-throughput analysis of solid injectable products in situ. In Figure 9, the vials are transported through a horizontal-bore superconducting magnet on a conveyer-like device, where samples stop when in the magnetic field for analysis. A similar approach can be used at low field, or an autosampler approach can also be implemented for the exchange of samples.

[0025] The disclosure relates to a method for determining product quality of a solid injectable product in situ, the method comprising: acquiring the1H NMR signal of the solid injectable product; andassessing the1H NMR signal properties as a measure of the quality of the solid injectable product.

[0026] As used herein, the term “assessing” generally refers to the analysis of signal from the injectable product. The1H NMR signal properties can be assessed either in the time domain by analysis of the free induction decay (FID) or by performing a Fourier transform of the signal to convert it from the time domain to the frequency domain. The display of data in the frequency domain is commonly referred to as a spectrum. The signal from the sample is most commonly referred to as a peak or combination of peaks. Peaks in the spectrum have unique characteristics related to their intensity, shape, and location in the spectrum. Location is often denoted by a chemical shift, but can also be represented by Hertz, or 1 / second. The properties of the peak may be complex depending on whether it is composed of multiple signals from mobile and immobile components. The assessment of the peaks can rely upon the analysis of one or more of these characteristics. For example, a simple determination of signal intensity may be sufficient for some samples, whereas a more complex analysis may incorporate analysis of one or more of peak intensity, peak shape, peak area (a combination of peak intensity and peak shape, often determined by integrating the peak), and peak location. Other factors can also be included in the analysis, including signal relaxation time or how quickly the signal returns to equilibrium. Signal relaxation time can be measured by using one or more experiments, such as a Carr Purcell Meiboom Gill sequence to measure spin-spin relaxation, often described as T2; an inversion recovery experiment or other experiment to measure spin-lattice relaxation, often described as T1; and a spin-locking experiment that can measure spin-lattice relaxation in the rotating frame, often described as Tirho.

[0027] Differences between samples can be defined as changes in peak intensities as a percentage or ratio, changes in peak breadth as a percentage or ratio, and changes in peak location as a percentage or ratio. Other characteristics, such as changes in relaxation time, may be assessed similarly. A threshold for acceptable versus non-acceptable samples, where the acceptance criteria could be related to mobility, stability, reconstitution time, or other critical quality attributes that define the acceptability of the sample, may be determined based upon the differences described above.

[0028] There are several ways in which the differences between samples can be assessed (e.g., identified and quantified) in the assessing step of the claimed methods. For example, assessing can include identifying a region of the spectrum, such as a peak, and determine if the intensity of that region exceeds a threshold,which would indicate if the sample is acceptable or would fail the analysis (poor quality). FIG. 10A is a plot of human serum albumin (HSA) percent monomer versus time at 50°C. It is clear from the plot that the stability of the HSA increases with increasing sucrose concentration. When there is no sucrose (100% protein, which is referenced as “100P” in FIG. 10A) the protein is unstable and degrades over 36 weeks at 50°C. The stability of the HSA begins to be high at approximately 70% HSA and 30% sucrose (70P30S) such that there is only a small amount of degradation after 36 weeks at 50°C. The stability of the HSA is consistently high at 50% sucrose, 50% HSA (50P50S) and samples containing higher ratios of sucrose to HSA. The data in FIG. 10A were obtained at various time points using the method described in Mol. Pharmaceutics 21: 3163-3172 (2024), which is incorporated by reference as if fully set forth herein and is a destructive method for analysis of the amount of degradation of a sample. Samples tested in this way cannot be used again for any further testing once the sample is reconstituted for analysis, which is one reason why the claimed methods are so powerful. In this destructive method, samples are stored in a desiccator in an incubator at 50°C for 36 weeks. Storage at 50°C provides accelerated stability conditions but is still significantly below the glass transition temperature of the disaccharides. At 0, 1 , 2, 4, 8, 18, and 36 weeks, three vials were removed from the incubator, and the relative amount of aggregation was measured by size-exclusion chromatography (SEC). A TOSOH TSK Gel G2000SWXL column (30 cm x 7.8 mm ID, 5 pm particle size) can be used for all SEC analyses. The column temperature can be set at 30°C during the analysis. A mobile phase of 50 mM potassium phosphate at pH 6.5 with 200 mM sodium sulfate and a flow rate of 0.5 mL / min can be used. The sample can be reconstituted with Milli-Q ultrapure water to a concentration of 1 mg / mL HSA and 20 pL can be injected onto the column. HSA peaks can be detected by UV absorbance at 254 nm.

[0029] FIG. 10B is spectra obtained for bovine serum albumin (BSA) and BSA / sucrose samples. From left to right, each sample contains increasing amounts of sucrose as shown. As was the case in FIG. 10A with 100% HSA (sample 1) one can see that the peak intensity is approximately 5000 units. And as was observed in FIG. 10A, the relative intensity of the peaks decrease with increasing amounts of sucrose until there is little deviation at about 50% BSA / 50% sucrose. In sum, the data for HSA, measured using a destructive method tracks the data obtained for BSA using the claimed methods. And the data obtained by the claimed method and shown in FIG. 10B has been validated using the destructive method in Mol. Pharmaceutics 21: 3163-3172 (2024). In the exampleshown in FIG. 10B, the assessing the1H NMR signal properties involves measuring the1H NMR signal intensity, where a change in signal intensity of about 25% or greater (higher or lower intensity, though in the case of FIG. 10B, the signal intensity increases as a function of the quality of the solid injectable product, with the lower quality sample being sample 1) can be interpreted as a measure of the quality of the solid injectable product. Thus, for example, sample 4 has a signal intensity of approximately 1000. Samples 5-10, on the other hand, have a signal intensity of approximately 750. The percent change in intensity is approximately 25%. Accordingly, in one example, a change in the signal intensity of about 20% or greater (e.g., about 30% or greater, about 40% or greater, about 50% or greater, about 75% or greater, about 125% or greater, about 200% or greater, about 300% or greater, about 500% or greater, about 750% or greater, or about 1000% or greater; or about 25% to about 1000%, about 25% to about 500%, about 25% to about 100%, about 25% to about 50% or about 25% to about 30%) can be interpreted as a measure of a solid injectable product having poor quality. Samples with changes of less than about 20% can be interpreted as a measure of a solid injectable product having acceptable quality.

[0030] Assessing can also include integrating the region of the1H NMR spectrum and determine if the integrated area exceeds a threshold value, which would indicate that the sample is acceptable or unacceptable, of good quality and poor quality, respectively. In the case of signal integration, one would be measuring a change in signal integration, such that, for example, a change in signal integration of about 20% or greater (higher or lower integration; e.g., about 30% or greater, about 40% or greater, about 50% or greater, about 75% or greater, about 125% or greater, about 200% or greater, about 300% or greater, about 500% or greater, about 750% or greater, or about 1000% or greater; or about 25% to about 1000%, about 25% to about 500%, about 25% to about 100%, about 25% to about 50% or about 25% to about 30%) can be interpreted as a measure of a solid injectable product having poor quality. Samples with changes of less than about 20% can be interpreted as a measure of a solid injectable product having acceptable quality.

[0031] Assessing can also include measuring signal breadth of a region of the1H NMR spectrum and determine if the signal breadth exceeds a threshold value, which would indicate that the sample is acceptable or unacceptable, of good quality and poor quality, respectively. In the case of signal breadth, one would be measuring a change in signal breadth, such that, for example, a change in signal breadth of about 20% or greater (higher or lower integration; e.g., about 30% or greater, about 40% or greater, about 50% or greater, about 75% or greater, about125% or greater, about 200% or greater, about 300% or greater, about 500% or greater, about 750% or greater, or about 1000% or greater; or about 25% to about 1000%, about 25% to about 500%, about 25% to about 100%, about 25% to about 50% or about 25% to about 30%) can be interpreted as a measure of a solid injectable product having poor quality. Samples with changes of less than about 20% can be interpreted as a measure of a solid injectable product having acceptable quality.

[0032] Assessing can also include measuring relaxation time, which could be the spin-lattice relaxation time (Ti), the spin-lattice relaxation time in the rotating frame (Tirho), and / or the spin-spin relaxation time (T2 or T2*) when obtaining a1H NMR spectrum, and determine if the spin-lattice relaxation time exceeds a threshold value, which would indicate that the sample is acceptable or unacceptable, of good quality and poor quality, respectively. In the case of spin-lattice relaxation time, one would be measuring a change in spin-lattice relaxation time, such that, for example, a change in spin-lattice relaxation time of about 20% or greater (higher or lower integration; e.g., about 30% or greater, about 40% or greater, about 50% or greater, about 75% or greater, about 125% or greater, about 200% or greater, about 300% or greater, about 500% or greater, about 750% or greater, or about 1000% or greater; or about 25% to about 1000%, about 25% to about 500%, about 25% to about 100%, about 25% to about 50% or about 25% to about 30%) can be interpreted as a measure of a solid injectable product having poor quality. Samples with changes of less than about 20% can be interpreted as a measure of a solid injectable product having acceptable quality.

[0033] A combination of the foregoing four methods of assessing (i.e., assessing changes in signal intensity, signal integration, signal breadth, and spin-lattice relaxation time) can also be used as a measure of a solid injectable product having poor quality. The binning of samples may also not be limited to simply poor or acceptable quality. There could be multiple bins reflecting a gradation in the properties of the samples. The identification and quantitation of differences can be used for several purposes. For example, one purpose would be to determine product acceptability if specific product quality attributes were identified. Another example would be to use these differences to identify samples for further study. For example, a sample that had an identifiable characteristic as defined earlier may be chosen for a subsequent stability study or other testing (e.g. water content assessment) to determine product quality.

[0034] Even though the solid injectable product may be solid, the solid injectable product may comprise solid and liquid components, where the liquid componentmay correspond to free water or other highly-mobile components that impact product quality. Examples of1H NMR signal properties of the solid injectable product that can assess product quality include the measurement of relaxation parameters, such as1H Ti ,1H Tirho, and / or1H T2.

[0035] The solid injectable product can be a lyophilized injectable product. Alternatively, the solid injectable product is a spray dried, foam dried, or produced by any other method known in the art to produce a solid injectable product. Alternatively, or in addition, the solid injectable product comprises a protein pharmaceutical product. Alternatively, the solid injectable product comprises a small molecule pharmaceutical product. Alternatively, the solid injectable product comprises a polynucleotide pharmaceutical product.

[0036] The assessing of the1H NMR signal properties can include correlating the mobility of the solid injectable product to injectable product stability. Alternatively, the assessing of the1H NMR signal properties can include correlating the solid injectable product mobility to injectable product quality. Alternatively, the assessing of the1H NMR signal properties can include evaluating a1H NMR signal intensity, and / or signal shape, and / or the Fourier transformed spectrum. The signal intensity and / or signal shape can be that of the1H NMR spectrum. Alternatively, the signal intensity and / or signal shape can be that of the free induction decay (FID). Thus, for example, a broad and intense1H NMR spectrum signal is indicative of low injectable product stability. Alternatively, or in addition, a slow free induction decay, for example, lasting more than 200 microseconds, lasting more than 500 microseconds before 90% signal decay, is indicative of low injectable product stability.

[0037] The disclosure also includes a system for predicting the quality of a solid injectable product in situ, the system comprising: a nuclear magnetic resonance (NMR) device comprising an NMR coil; and a device for introducing the solid injectable product in a container into an NMR coil. An example of such a system is provided in FIG. 9. An example of the device for introducing the solid injectable product in a container into an NMR coil is shown in FIG. 2. The system allows for performing the methods described herein such that an analysis of the solid injectable product in situ can be performed for the purpose of evaluating product quality. The system can be a high-throughput system. For example, the system can evaluate a plurality of containers at a rate of at least 10 containers per hour; at a rate of at least 60 containers per hour; or at a rate of at least 120 containers per hour.

[0038] Values expressed in a range format should be interpreted in a flexible manner to include not only the numerical values explicitly recited as the limits of the range, but also to include all the individual numerical values or sub-ranges encompassed within that range as if each numerical value and sub-range were explicitly recited. For example, a range of “about 0.1 % to about 5%” or “about 0.1 % to 5%” should be interpreted to include not just about 0.1 % to about 5%, but also the individual values (e.g., 1 %, 2%, 3%, and 4%) and the sub-ranges (e.g., 0.1 % to 0.5%, 1.1% to 2.2%, 3.3% to 4.4%) within the indicated range. The statement “about X to Y” has the same meaning as “about X to about Y,” unless indicated otherwise. Likewise, the statement “about X, Y, or about Z” has the same meaning as “about X, about Y, or about Z,” unless indicated otherwise.

[0039] In this document, the terms “a,” “an,” or “the” are used to include one or more than one unless the context clearly dictates otherwise. The term “or” is used to refer to a nonexclusive “or” unless otherwise indicated. In addition, it is to be understood that the phraseology or terminology employed herein, and not otherwise defined, is for the purpose of description only and not of limitation. Any use of section headings is intended to aid reading of the document and is not to be interpreted as limiting. Further, information that is relevant to a section heading can occur within or outside of that particular section. Furthermore, all publications, patents, and patent documents referred to in this document are incorporated by reference herein in their entirety, as though individually incorporated by reference. In the event of inconsistent usages between this document and those documents so incorporated by reference, the usage in the incorporated reference should be considered supplementary to that of this document; for irreconcilable inconsistencies, the usage in this document controls.

[0040] In the methods described herein, the steps can be carried out in any order without departing from the principles of the invention, except when a temporal or operational sequence is explicitly recited. Furthermore, specified steps can be carried out concurrently unless explicit claim language recites that they be carried out separately. For example, a claimed step of doing X and a claimed step of doing Y can be conducted simultaneously within a single operation, and the resulting process will fall within the literal scope of the claimed process.

[0041] The term “about” as used herein can allow for a degree of variability in a value or range, for example, within 10%, within 5%, or within 1 % of a stated value or of a stated limit of a range.

[0042] The term “substantially” as used herein refers to a majority of, or mostly, as in at least about 50%, 60%, 70%, 80%, 90%, 95%, 96%, 97%, 98%, 99%, 99.5%, 99.9%, 99.99%, or at least about 99.999% or more.

[0043] The term “substantially no” as used herein refers to less than about 30%, 25%, 20%, 15%, 10%, 5%, 3%, 2%, 1 %, 0.5%, 0.1 %, 0.05%, 0.001 %, or at less than about 0.0005% or less or about 0% or 0%.

[0044] Those skilled in the art will appreciate that many modifications to the embodiments described herein are possible without departing from the spirit and scope of the present disclosure. Thus, the description is not intended and should not be construed to be limited to the examples given but should be granted the full breadth of protection afforded by the appended claims and equivalents thereto. In addition, it is possible to use some of the features of the present disclosure without the corresponding use of other features. Accordingly, the foregoing description of or illustrative embodiments is provided for the purpose of illustrating the principles of the present disclosure and not in limitation thereof and can include modification thereto and permutations thereof.

[0045] The disclosure also relates to the following numbered Embodiments, which are listed in no particular order of importance:1. A method for determining product quality of a solid injectable product in situ, the method comprising: acquiring the1H NMR signal of the solid injectable product; and assessing the1H NMR signal properties as a measure of the quality of the solid injectable product.2. The method of Embodiment 1 , wherein the1H NMR signal properties of the solid injectable product are used to measure relaxation parameters, such as1H Ti,1H Tirho, and / or1H T2.3. The method of Embodiment 1 , wherein the solid injectable product is a lyophilized injectable product.4. The method of Embodiment 1 , wherein the solid injectable product is a spray dried or foam dried.5. The method of Embodiment 1 , wherein the solid injectable product comprises a protein pharmaceutical product.6. The method of Embodiment 1 , wherein the solid injectable product comprises a small molecule pharmaceutical product.7. The method of Embodiment 1 , wherein the solid injectable product comprises a polynucleotide pharmaceutical product.8. The method of Embodiment 1 , wherein the assessing comprises correlating the mobility of the solid injectable product to injectable product stability.9. The method of Embodiment 1 , wherein the assessing comprises correlating the solid injectable product mobility to injectable product quality.10. The method of Embodiment 1 , wherein the assessing comprises evaluating a1H NMR signal intensity, and / or signal shape, and / or the Fourier transformed spectrum.11. The method of Embodiment 9, wherein the signal intensity and / or signal shape is that of the1H NMR spectrum.12. The method of Embodiment 9 or 10, wherein the signal intensity and / or signal shape is that of the free induction decay (FID).13. The method of Embodiment 1 , wherein a broad and intense1H NMR spectrum signal is indicative of low injectable product stability.14. The method of Embodiment 1 , wherein a long free induction decay (FID) is indicative of low injectable product stability.15. The method of Embodiment 1 , wherein the assessing comprises assessing changes in signal intensity, signal integration, signal breadth, and / or spin-lattice relaxation time.16. The method of Embodiment 15, wherein a change in signal integration, signal breadth, and / or spin-lattice relaxation time of about 20% or greater is a measure of a solid injectable product having poor quality.17. The method of Embodiment 15, wherein a change in signal integration, signal breadth, and / or spin-lattice relaxation time of about 25% to about 1000% is a measure of a solid injectable product having poor quality.18. The method of Embodiment 15, wherein a change in signal integration, signal breadth, and / or spin-lattice relaxation time of less than about 20% is a measure of a solid injectable product having acceptable quality.19. The method of Embodiment 1 , wherein the assessing comprises assessing changes in signal intensity.20. The method of Embodiment 19, wherein a change in signal integration of about 20% or greater is a measure of a solid injectable product having poor quality.21. The method of Embodiment 19, wherein a change in signal integration of about 25% to about 1000% is a measure of a solid injectable product having poor quality.22. The method of Embodiment 19, wherein a change in signal integration of less than about 20% is a measure of a solid injectable product having acceptable quality.23. The method of Embodiment 1 , wherein the assessing comprises assessing changes in signal integration.24. The method of Embodiment 1 , wherein the assessing comprises assessing changes in signal breadth.25. The method of Embodiment 1 , wherein the assessing comprises assessing changes in spin-lattice relaxation time.26. A system for predicting the quality of a solid injectable product in situ, the system comprising: a nuclear magnetic resonance (NMR) device comprising an NMR coil; and a device for introducing a solid injectable product in a container into an NMR coil.27. The system of Embodiment 26, wherein the system is a high-throughput system.28. The system of Embodiment 26, wherein the system evaluates a plurality of containers at a rate of at least 10 containers per hour.29. The system of Embodiment 26, wherein the system evaluates a plurality of containers at a rate of at least 60 containers per hour.30. The system of Embodiment 26, wherein the system evaluates a plurality of containers at a rate of at least 120 containers per hour.Examples

[0046] The disclosure can be better understood by reference to the following examples which are offered by way of illustration. The disclosure is not limited to the examples given herein.Example 1 : Water Standards

[0047] A sealed sample containing ~10 pL water was prepared. The sealed sample was placed in a 2 mL vial. The1H NMR spectrum 102 of the sample (FIG. 1) was obtained (50 acquisitions, 4 second repetition delay; measurement time ~4 min; high signal-to-noise ratio). The1H NMR spectrum was taken using a specially designed probe having an elevated coil designed to observe the lyophilized cake and to avoid background signals arising from the vial stopper and rest of the NMR probe (FIG. 2). In FIG. 2, the lyophilized sample is contained in vial 202 comprising a vial stopper and an aluminum cap 204 placed on elevated coil 206. A copper cap (not shown), similar to a cup with a diameter of ~18 mm and a length of ~12 mm, is placed over the sealed vial such that it shields the cap from the RF field to provide additional background signals arising from the vial stopper.

[0048] A1H NMR spectrum was also obtained for an empty vial without a stopper 104 (1000 acquisitions, 1 second repetition delay); and for an empty vial with a stopper 106 (1000 acquisitions, 1 second repetition delay). For the latter spectrumthe copper cap was used to minimize background signals arising from the vial stopper.Example 2: 20% Sucrose Formulations

[0049] Three identical 20% sucrose samples were prepared that contained 100 mg (total solids) in a 2 mL vial with 0.5 mL fill volume. Other samples prepared in the same lyophilization cycle had a water content of 3.47% ± 0.18 as measured by Karl Fischer titration of other vials prepared in the same cycle. Samples were stored at -20 °C after lyophilization prior to analysis. The1H NMR spectra of each sample were taken using the same probe set up as described in Example 1. The1H NMR spectra of each sample is shown in FIG. 3. Another set of1H NMR spectra were taken three days later (FIG. 4) after samples were stored at ambient conditions. Even though the samples were prepared to be otherwise identical, it is clear that sample 2 is different, since it has a1H NMR spectrum where the sole peak is more intense that the spectra for samples 1 and 3 (see FIG. 5 for side-by- side comparison). The more intense peak is indicative of water that is more mobile, which, in turn, is indicative of a less stable product. The NMR spectra are consistent with the visual appearance of the cakes, where the smallest signal corresponds to the largest cake (sample 1), and the largest signal corresponds to the smallest cake (sample 2).Example 2: 65% Trehalose:35% Bovine Serum Albumin (BSA) Formulations

[0050] In this experiment, 320 identical 65% trehalose:35% BSA samples were prepared and lyophilized, that contained 25 mg (total solids; 8.75 mg BSA and 16.25 mg trehalose) in a 2 mL vial with 0.5 mL fill volume. Analysis of vials in the vicinity of the samples tested showed that they had a water content of ~1 .6-2.2% using Karl Fischer Titration of other vials prepared in the same cycle. Four samples were randomly selected from the distribution shown in FIG. 6, where each rectangle represents a sample and its location in a lyophilizer. Samples that are closer to the edges will dry more quickly and are expected to have lower moisture content.

[0051] FIG. 7 shows the1H NMR spectra of each of the samples tested, 1-4. Though the cakes appear very similar from visual inspection, the spectrum of sample 4 shows that the sample has a higher unbound water content and is of poor quality relative to samples 1-3.Example 4: Time Domain NMR (TDNMR) for Sample Stability on 20% Sucrose Formulations

[0052] Solid-state TD-NMR experiments were performed on a MQ20 Minispec (Bruker, Billerica, MA; B0(1 H) = 20 MHz). All TD-NMR experiments were performed using a Bruker Absolute 18 mm probe).

[0053] Data collection was obtained at a nominal temperature of 18°C. Data collection was acquired using the solid echo fast relaxation experiment. Data were collected using a 0.3 s recycle delay, receiver gain of 102 dB, 3000 scans, magnitude detection mode and an acquisition time of 1 ms.

[0054] Four samples were prepared using 20% sucrose (total solids: 100 mg; 2 mL vial with 0.5 mL fill volume; 3.47% ± 0.18 MC). The samples were prepared similarly to those described in Example 2. The free induction decay (FID) was evaluated for each sample: intensity decay as a function of time. It was found that if the sample had a FID that decayed to close to 0 on the order of 1 ms (FIG. 8A) was indicative of water in the sample that is highly mobile, which, in turn, is indicative of a less stable product. The other three samples had a Fl D that decayed to close to 0 on the order of 0.1 ms as shown in FIGS. 8B-8D. FIG. 9 shows images of the four samples. The trends of cake size vs. signal intensity for the TD-NMR data follow the trends observed in Example 2.

Claims

What is claimed is:

1. A method for determining product quality of a solid injectable product in situ, the method comprising: acquiring the1H NMR signal of the solid injectable product; and assessing the1H NMR signal properties as a measure of the quality of the solid injectable product.

2. The method of claim 1 , wherein the1H NMR signal properties of the solid injectable product are used to measure relaxation parameters, such as1H Ti,1H Tirho, and / or 1H T2.

3. The method of claim 1 , wherein the solid injectable product is a lyophilized injectable product.

4. The method of claim 1 , wherein the solid injectable product is a spray dried or foam dried.

5. The method of claim 1 , wherein the solid injectable product comprises a protein pharmaceutical product.

6. The method of claim 1 , wherein the solid injectable product comprises a small molecule pharmaceutical product.

7. The method of claim 1 , wherein the solid injectable product comprises a polynucleotide pharmaceutical product.

8. The method of claim 1 , wherein the assessing comprises correlating the mobility of the solid injectable product to injectable product stability.

9. The method of claim 1 , wherein the assessing comprises correlating the solid injectable product mobility to injectable product quality.

10. The method of claim 1 , wherein the assessing comprises evaluating a1H NMR signal intensity, and / or signal shape, and / or the Fourier transformed spectrum.

11. The method of claim 9, wherein the signal intensity and / or signal shape is that of the1H NMR spectrum.

12. The method of claim 9 or 10, wherein the signal intensity and / or signal shape is that of the free induction decay (FID).

13. The method of claim 1 , wherein a broad and intense1H NMR spectrum signal is indicative of low injectable product stability.

14. The method of claim 1 , wherein a long free induction decay (FID) is indicative of low injectable product stability.

15. The method of claim 1, wherein the assessing comprises assessing changes in signal intensity, signal integration, signal breadth, and / or spin-lattice relaxation time.

16. The method of claim 15, wherein a change in signal integration, signal breadth, and / or spin-lattice relaxation time of about 20% or greater is a measure of a solid injectable product having poor quality.

17. The method of claim 15, wherein a change in signal integration, signal breadth, and / or spin-lattice relaxation time of about 25% to about 1000% is a measure of a solid injectable product having poor quality.

18. The method of claim 15, wherein a change in signal integration, signal breadth, and / or spin-lattice relaxation time of less than about 20% is a measure of a solid injectable product having acceptable quality.

19. The method of claim 1, wherein the assessing comprises assessing changes in signal intensity.

20. The method of claim 19, wherein a change in signal integration of about 20% or greater is a measure of a solid injectable product having poor quality.

21. The method of claim 19, wherein a change in signal integration of about 25% to about 1000% is a measure of a solid injectable product having poor quality.

22. The method of claim 19, wherein a change in signal integration of less than about 20% is a measure of a solid injectable product having acceptable quality.

23. The method of claim 1, wherein the assessing comprises assessing changes in signal integration.

24. The method of claim 1, wherein the assessing comprises assessing changes in signal breadth.

25. The method of claim 1, wherein the assessing comprises assessing changes in spin-lattice relaxation time.

26. A system for predicting the quality of a solid injectable product in situ, the system comprising: a nuclear magnetic resonance (NMR) device comprising an NMR coil; and a device for introducing a solid injectable product in a container into an NMR coil.

27. The system of claim 26, wherein the system is a high-throughput system.

28. The system of claim 26, wherein the system evaluates a plurality of containers at a rate of at least 10 containers per hour.

29. The system of claim 26, wherein the system evaluates a plurality of containers at a rate of at least 60 containers per hour.

30. The system of claim 26, wherein the system evaluates a plurality of containers at a rate of at least 120 containers per hour.

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