Apparatus and method for solid phase extraction

Abstract

To provide a device and method for optimizing solid phase extraction (SPE) purification conditions for the isolation of a compound from a composition containing the compound, preferably from a crude reaction mixture.SOLUTION: A cassette comprises: (i) a flowpath comprising a first end and a second end; and (ii) a plurality of valves arranged along the flowpath, where each of the valves is selectively fluidly connected to one of a number of components. The components comprise: (a) 1-5 composition vials; (b) 1-3 SPE cartridges; (c) 4-10 solvent vials; (d) a water vial; and (e) a transfer line.SELECTED DRAWING: Figure 1

Description

[Technical Field]

[0001] The present invention relates to the field of solid phase extraction (SPE). More particularly, the present invention relates to SPE conditions The present invention is directed to an apparatus and method for optimizing the [Background technology]

[0002] Automated synthesis systems are important for the production of radiopharmaceuticals. Synthesis systems such as the GE Healthcare (GE Healthcare) GE Healthcare Synthesizer are being developed for clinical applications. The FASTlab synthesizer provides dose generation. Examples of such radiopharmaceuticals include: 18 F-FLT ([ 18 F]fluorothymidine), 18 F-FDDNP(2-(1-{6-[(2-[ 1 8 F]fluoroethyl)(methyl)amino]2-naphthyl}ethylidene)malononitrile ), 18 F-FHBG(9-[4-[ 18 F]Fluoro-3-(hydroxymethyl)butyl le]guanine or [ 18 F]-penciclovir), 18 F-FESP([ 18 F]-F difluoroethylspiperone), 18 Fp-MPPF(4-(2-methoxyphenyl)-1 -[2-(N-2-pyridinyl)-p-[ 18 F]Fluorobenzamido]ethylpipera Gin) and 18 F-FDG([ 18 F]-2-Deoxy-2-fluoro-D-glucose This includes:

[0003] Such synthesis systems / devices include a flow channel including a first end and a second end; and The cassette is used with a plurality of valves arranged along the flow path, and the plurality of valves Each of the valves is selectively fluidly connected to one of several components, typically , the process of producing a reagent vial, a compound, to synthesize a specific radioactive tracer Reactors, cartridges, filters, syringes, and dispensers in which one or more reactions are carried out. The different radiopharmaceuticals have different capacities for that radiopharmaceutical. The device is made using a customized dedicated cassette. To achieve this, operate the stopcock and syringe to release the radioisotope. and a component cooperating with said component to enable a feed fluid having The synthesis system is also configured to effectively engage any of the components required for the chemical reaction. The first reactor receives one or more (e.g., two) heating The cavity / heating element may be included.

[0004] The synthesis system operates the required pumps, syringes, valves, and heating elements. It is programmed to also direct the source fluids to mix with the reagents and to The chemical reaction is carried out through the reservoir, and the output tracer and waste fluid are separated into appropriate selectively pumping into the receptacle to obtain final product, waste, etc. The delivery of motive gas (e.g., nitrogen) and application of vacuum are controlled. The extracted fluid is typically input into another system for either purification and / or dispensing. However, synthetic systems may also be purified (or partially purified) for further processing. Connect to or include a separate purification system that returns the (removed) compound to the system. can be done.

[0005] Such an automated synthesizer of the prior art is described in WO 2007 / 042781. It is stated in the

[0006] Existing synthesis systems, such as the FASTlab synthesizer, include some form of purification step. Although a system may be included, the system is not designed to optimize purification conditions or It is not optimized and is not used in prior art (e.g., FASTlab system). Such purification methods are currently being optimized using existing SPE manifold systems. In this case, SPE is performed manually. This method is useful for determining the appropriate purification method, as it involves one or more purification parameters that are Moreover, once suitable purification conditions are identified, the resulting purification method is Further adaptation and optimization will be required to make it compatible with the production system.

[0007] There are many examples of SPE robots available on the market. A key advantage is the ability to rapidly develop SPE purification methods using already existing production equipment. The time it takes to transfer an SPE method to an existing device (e.g., FASTlab) The gap is effectively eliminated because the underlying device may be the same in both cases.

[0008] Existing SPE robots can handle larger fractions than are mostly used in this invention. The ability to collect smaller fractions of the eluate for analysis is This is an improvement on the current SPE robot. The larger the fraction collected, the greater the impurity / product ratio. This reduces the inference about where the compound is eluting.

[0009] Furthermore, existing SPE robots occupy a large footprint in the laboratory. If your lab already has an automated synthesis system, some of the additional equipment dedicated to SPE optimization may not be necessary. This brings the advantage of

[0010] For example, the cassette of the present invention can accommodate up to six different mobile phases in one sequence. A FASTlab system programmed to assess three SPE cartridges This also has the advantage that the cartridges can be reused. This allows them to be repeatedly tested, saving both time and cartridge costs. savings are made in terms of

[0011] The process of optimizing product purification is designed to shorten the timescales as described above. It would be beneficial if it could be developed and compatible with current synthesizer systems. Summary of the Invention

[0012] The present invention provides a method for isolating a compound from a composition containing the compound, preferably a crude reaction mixture. The present invention provides a device for optimizing SPE purification conditions to isolate a compound from a composition. The compound to be tested may be any radioactive or non-radioactive compound, preferably a radioactive compound. The device of the first aspect of the present invention is described as a cassette. The set may be a separate piece of equipment and / or may be fitted into an already existing device. It may be part of a device and replace part of the original part. [Brief explanation of the drawings]

[0013] [Figure 1] FIG. 1 shows an example embodiment of a cassette of the present invention containing three SPE cartridges and six solvent vials (labeled mobile phase). [Figure 2] FIG. 1 shows an example embodiment of a cassette of the present invention containing two SPE cartridges and eight solvent vials (labeled mobile phase). [Figure 3] FIG. 1 shows an example embodiment of a cassette of the present invention containing one SPE cartridge and ten solvent vials (labeled mobile phase). [Figure 4] FIG. 1 shows an example of a cassette typically used in the FASTlab system for comparison with the present invention. [Figure 5] FIG. 1 shows the results of an embodiment of the process of the present invention applied to hydroxy impurities in flurpiridaz crude product. [Figure 6] 6 shows the results of an embodiment of the method of the present invention applied to a crude reaction mixture of flurpiridaz. In FIG. 6, the x-axis is volume (1-41 ml) and the y-axis is μg (0-25). DETAILED DESCRIPTION OF THE INVENTION

[0014] In a first aspect, the present invention provides a method for determining optimized solid phase extraction (SPE) purification conditions. A cassette of: (i) a flow channel including a first end and a second end; (ii) a plurality of valves arranged along the flow path, A plurality of valves, each of which is selectively fluidly connected to one of several components. With Lub and wherein said components include: (a) 1 to 5 vials of the composition; (b) 1–3 SPE cartridges; (c) 4–10 solvent vials; (d) a water vial; and (e) Transfer Line A cassette is provided, comprising:

[0015] The cassette of the present invention has a flow path including a first end and a second end. , particularly suitable for transporting fluids such as solvents and compositions, e.g., crude reaction mixtures. The composition vial may be a crude reaction mixture vial, a single reference standard vial, or It may be a reference standard mixture vial. A "crude reaction mixture" is a mixture containing one or more desired products. A "single reference standard" may contain a single impurity, e.g. For example, it may be an impurity that is commonly produced along with the desired product. "Mixture" refers to a mixture of impurities, e.g., impurities that are typically produced together with the desired product. It can also be an object.

[0016] "Fluidly connected" refers to the transfer of fluid to and from the vial (optionally) This indicates that the valve can be passed to other parts of the cassette. may be a three-way valve with three ports, and of the three associated ports This means that any two ports can be fluidly connected to each other while the third port is fluidly isolated. Suitable valves may also be stopcock valves, including rotatable stopcocks. may be.

[0017] The cassettes of the present invention may be used in systems or devices for synthesizing compounds for which the cassettes of the present invention are provided. The present invention may be coupled to or compatible with a variety of devices, such as the known FASTlab system. The set is compatible with the synthesizer system used for radiopharmaceutical synthesis as described above, and is SPE It is specifically designed to optimize the purification conditions. against the same and / or compatible systems used in the manufacture of This allows for the determination of an optimized SPE purification process for the product. This provides clear advantages over existing methods. For example, the purification process allows for the production of the desired product. Optimized for systems that are the same or compatible with those used in manufacturing ,In contrast to existing methods, further adaptations are required for the purification process to work in that system. is not necessary.

[0018] The cassette allows for the combination of SPE cartridges with different solvents (different chemical compositions or The solvent vials provide either a 100% or a different concentration of the solvent, while the SPE and solvent Alternative parameters of the solvent are available and can be combined to determine the optimal purification process. The present invention is designed to provide a device that

[0019] Preferably, the cassette comprises: (i) 3 SPE cartridges and 6 solvent vials; or (ii) two SPE cartridges and eight solvent vials; or (iii) 1 SPE cartridge and 10 solvent vials Includes.

[0020] One of the solvent vials is used to condition the SPE cartridge(s). The preparation can be carried out by passing 100% of the solvent through the SPE cartridge, followed by water. This can be done by passing it through a PE cartridge. The number of solvent vials used was 5 for 3 SPE cartridges and 2 for 7 solvent vials per SPE cartridge and 9 vials per SPE cartridge It can be considered a solvent vial.

[0021] In one embodiment, the cassette of the present invention contains a radioisotope, e.g., 18 [F] fluoride For example, the cassette does not contain any means for processing. cartridges,” e.g., nuclear reactions 18 O(p, n) 18 When the aqueous solution from F passes through, 18 Hold F, 18 It does not include an SPE cartridge through which O is passed. Ion exchange cartridges include anion exchange cartridges, e.g., quaternary methylammonium These include QMA cartridges, which are typically used for the radiosynthesis of radiolabeled compounds. In one embodiment, the cassette also includes a "positive In this case, the cationic counterion is not included in the cryptand. or a positively charged counterion such as a metal complex of a tetraalkylammonium salt, or It may also be a large but soft metal ion such as bidium or cesium.

[0022] In one embodiment, the cassette does not include a reactor.

[0023] The cassette may have 25 valves in a linear array. It has five valves and the cassette holds three SPE cartridges and six solvent vials. If it contains (i) 1 to 5 composition vials are 2, 12, 13, 14, and / or 16 vials wherein the valve is fluidly connected to the valve, and preferably up to three composition vials are used; (ii) Three SPE cartridges, preferably at valves 18, 20 and 22. fluidly connected to the valve; (iii) Six solvent vials flow through the 4th, 5th, 7th, 8th, 9th and 10th valves. Body connected; (iv) A water vial is in fluid connection with the 15th valve.

[0024] The cassette has a linear array of 25 valves, and the cassette contains two SPE cartridges and and 8 solvent vials, (i) 1 to 5 composition vials are 2, 12, 13, 14, and / or 16 vials wherein the valve is fluidly connected to the valve, and preferably up to two composition vials are used; (ii) Two SPE cartridges are preferably connected to the 20th and 22nd valves. Body connected; (iii) Eight solvent vials, numbered 4, 5, 7, 8, 9, 10, 17, and 18 is in fluid communication with the valve; (iv) A water vial is in fluid connection with the 15th valve.

[0025] The cassette has a linear array of 25 valves, and the cassette contains one SPE cartridge and and 10 solvent vials, (i) 1 to 5 composition vials are 2, 12, 13, 14, and / or 16 vials and wherein a maximum of one composition vial is used; (ii) One SPE cartridge is preferably in fluid communication with the 22nd valve. and; (iii) 10 solvent vials are placed at 4, 5, 7, 8, 9, 10, 17, 18, and 19 and is fluidly connected to the 20th valve; (iv) A water vial is in fluid connection with the 15th valve.

[0026] The cassette above is: (v) a syringe, preferably a 1 mL syringe, in fluid communication with the third valve; and; (vi) A syringe, preferably a 7 mL syringe, in fluid communication with the 11th valve. Nji and; (vii) A syringe, preferably a 7 mL syringe, in fluid communication with the 24th valve. With Linzi It may further include:

[0027] The cassette above is: (viii) The piping connecting the 1st and 25th valves It may further include:

[0028] The cassette above is: (ix) From the SPE cartridge(s) fluidically connected to the 23rd valve Output port for the eluate from It may further include:

[0029] In a second aspect of the present invention, the present invention provides an optimized SPE for a compound from a composition. 1. A method for determining purification conditions, comprising: (i) providing a cassette as defined in accordance with the first aspect of the present invention; and; (ii) a cassette containing a composition of compound in each of said 1 to 5 composition vials; or adding such a composition to each of said 1 to 5 composition vials. Steps and; (iii) transferring an aliquot of said composition to each of said 1 to 3 SPE cartridges; and passing the (iv) an aliquot of solvent from at least four of said four to ten solvent vials; passing the specific combination of The solvent in each of the 4 to 10 solvent vials may be a different solvent or a different concentration. and passing the solvent through the same solvent; (v) Dissolve the compound to be purified from the SPE cartridge or cartridges. Step of issuing; (vi) evaluating the eluted product of step (v); (vii) The eluted product of step (v) from each cartridge and each solvent. determining optimized purification conditions by comparing; The present invention provides a method comprising:

[0030] The at least one solvent is: (i) ethanol, (ii) methanol, (iii) acetonitrile, nitrile, or any alternative organic solvent known in the art, or a combination thereof If a different solvent is used, it may be necessary to include an aqueous solvent exchange step. Therefore, the product is preferably eluted with ethanol or aqueous ethanol.

[0031] The method involves preparing one to three SPE cartridges as the first step in the process. The cartridge may further comprise a step of: The solution may be adjusted with either ethanol or acetonitrile, followed by water. The volumes of organic solvent and water used can be varied to suit different cartridges. For example, 7 mL of organic solvent and 7 mL of water may be suitable. Smaller organic solvent volumes, e.g., 2 mL, may be suitable, in which case multiple cartridges may be used. The entire binder can be prepared using solvent from just one syringe. Use a syringe filled with water to ensure complete removal of the solvent. The crude product is then loaded onto the SPE cartridge(s).

[0032] Each cartridge is then washed with a mobile phase eluent in variable volume fractions, where The product is "more detail" or smaller volume and "analysis time" or collected volume. It is a compromise between the smaller volume and the need to analyze more sample. The eluate fractions are collected and analyzed, for example, by analytical HPLC. The cartridge is cleaned with 100% organic solvent and then mixed with different mobile phases during the process. The composition can be reconditioned so that it is ready for use again, reducing operator processing time. To minimize this, samples were collected in a 96-well plate and injected into an autosampler. The collected samples can be analyzed overnight using an HPLC system. The results can then be read the following morning. The evaluation in step (vi) can be carried out by any suitable method. This may be carried out using, for example, HPLC, LC-MS or TLC.

[0033] 1, 2, 3, 4 or 5 composition vials may be used. In particular, 1, 2 or 3 Preferably, the number of composition vials is equal to the number of vials in the SPE cartridge. It may be the same as the number of cartridges.

[0034] The term "eluting" refers to the process of releasing one or more selected compounds bound to a solid phase. Elution refers to passing a solution through an SPE cartridge. by passing it through a trough and then through a transfer line for collection Preferably, collection is in a 96-well plate. The chemical properties of a given compound act to release it from the SPE cartridge. and depending on the nature of the SPE column chemistry, e.g., organic, acidic, or basic solvents. The solvent may be a solvent.

[0035] The above method may further include a step of eluting impurities. , which may be carried out before and / or after step (v) above to elute the desired product. Preferably, the elution of impurities occurs before and after step (v) above, which elutes the desired product. It may be executed later.

[0036] The product obtained from the composition is at least 95%, preferably at least 97%, more preferably The purification process is ideally effective when obtained with a purity of preferably at least 99%. be.

[0037] The present invention also provides a compound purified according to the second aspect of the invention.

[0038] In a third aspect, the present invention provides a method for producing a medicament for the treatment of a pulmonary arthritis, comprising: (i) a cassette according to the first aspect; (ii) 1 to 5 vials of the composition; (iii) 1–3 SPE cartridges; (iv) 4–10 solvent vials; (v) a water vial; and (vi) Transfer lines A kit comprising:

[0039] Figures 1 to 6 are provided to illustrate the present invention in a non-limiting manner. [Example]

[0040] The following examples illustrate the invention in a non-limiting manner:

[0041] The first experiment consisted of 20, 30, and 40% ethanol in 0.1% aqueous formic acid. The mobile phase was used to carry out the analysis using a crude sample containing compound GE-179. The structure of 9 is as follows:

[0042] [ka]

[0043] The FASTlab cassette was set up as shown in Figure 1. All three cartridges was prepared with 100% ethanol (2 mL) followed by 100% water (7 mL). The product (dissolved in 10% ethanol and 90% water) was loaded onto 3 x tC18 cartridges. The first cartridge was filled with 20% ethanol, the second with 30% ethanol, and the third with 3 was washed with 40% ethanol (18 mL was passed through each column, and 1 mL fractions were collected). 54 samples were collected in a 96-well plate and analyzed using an analytical HPLC system equipped with an autosampler. The data were analyzed using the C system. The maximum yield was obtained using 40% EtOH. All of the eluate was eluted from the cartridge in the first 6 mL, but when 20% was used, it took 18 mL of washing time. After cleaning, the desired product and all subsequent eluting peaks were captured on the cartridge. From these results, it was found that the optimum condition for cleaning the cartridge is 20% or less. The optimum selective ethanol concentration for the product is found to be higher but lower than 40%. Further experiments using the same techniques are needed to determine the optimal dissolution.

[0044] Experiments were also carried out directed to the purification of flurpiridaz, which has the following structure:

[0045] [ka]

[0046] The main impurity in the flurpiridaz crude product is a hydroxy impurity, which has the following structure: Has:

[0047] [ka]

[0048] The FASTlab cassette was set up as shown in Figure 2. 100% ethanol (7 Both cartridges were conditioned with 100% water (7 mL). Pure material (dissolved in 1:10 ethanol:water) was loaded onto 2 x tC18 cartridges. The first cartridge was washed with 40% acetonitrile and the second with 35% ethanol. (41 mL was passed through each column, and 1 mL fractions were collected.) 82 samples were placed in a 96-well plate. The extracts were collected in a tray and analyzed using an analytical HPLC system equipped with an autosampler.

[0049] Figure 5 shows that the hydroxy impurity is reduced at 40% acetone compared to 35% ethanol (solid line). The results show that nitrile (dotted line) elutes as a tighter band. From the results, 40% acetonitrile was better than 35% ethanol, and the crude product It was determined that a wash volume of 14–21 mL was sufficient to remove the major impurities. The line in Figure 5 corresponding to the 35% ethanol elution contains a sharp drop, which is Note that this is due to an incorrect data point.

[0050] The experiment was repeated using the crude reaction mixture of flurpiridaz:

[0051] The FASTlab cassette was set up as shown in Figure 3. The cartridge was The crude product (approximately 2 mL) was diluted with ethanol (7 mL) and then 100% water (7 mL). 0% acetonitrile in 80% water) was loaded onto a C18 cartridge. The cartridge was washed with 40% acetonitrile (41 mL, 1 mL fractions were collected). 41 samples were collected in a 96-well plate and analyzed using an autosampler equipped HP analytical system. The LC system was used to analyze the hydroxyl standard. From these results shown in Figure 6, it is clear that the hydroxy impurities can be removed without eluting the product. It was confirmed that a wash volume of approximately 16 mL was sufficient to remove the other compounds present in very small amounts. The impurities were cyano and chloro impurities.

[0052] The embodiments of the invention described herein are likely to be of broad utility and application. It will be readily understood by those skilled in the art that the present invention will now be described with reference to exemplary embodiments. Although described in detail herein, this disclosure is illustrative and is an example of an embodiment. It is understood that this is provided to provide a possible disclosure of exemplary embodiments. The disclosure is not intended to be construed as limiting the embodiments of the present invention. without regard to any other such embodiments, adaptations, variations, modifications, equivalent arrangements, or the like. The scope of the present invention is defined by the appended claims. It has been done.

Claims

1. 1. A method for determining SPE purification conditions for the isolation of a compound from a composition, said method comprising: (i) providing a cassette, said cassette comprising: (a) a flow channel including a first end and a second end; (b) a plurality of valves disposed along the flow path, each of the plurality of valves being selectively fluidly connected to one of several components; wherein the component comprises: (1) 1 to 5 composition vials; (2) 1 to 3 SPE cartridges; (3) 4 to 10 solvent vials; (4) a water vial; and (5) Transfer line providing a cassette comprising: (ii) the cassette contains a composition of the compound in each of the one to five composition vials or adds the composition to each of the one to five composition vials; (iii) passing an aliquot of said composition through each of said 1 to 3 SPE cartridges; (iv) passing specific combinations of aliquots of solvents from at least four of the four to ten solvent vials through one or more SPE cartridges, wherein the solvents in each of the four to ten solvent vials are either different solvents or different concentrations of the same solvent; (v) eluting the compound to be purified from the or each SPE cartridge; (vi) evaluating the eluted product of step (v); (vii) determining the SPE purification conditions by comparing the eluted product of step (v) from each cartridge and each solvent; A method comprising:

2. １８ 10. The method of claim 1, which does not include any means for treating fluorides of F.

3. The cassette (i) 3 SPE cartridges and 6 solvent vials; or (ii) two SPE cartridges and eight solvent vials; or (iii) 1 SPE cartridge and 10 solvent vials 3. The method of claim 1 or claim 2, comprising:

4. In (i) of claim 3, the cassette has a linear array of 25 valves; and (i) the 1 to 5 composition vials are fluidly connected to the 2, 12, 13, 14, and / or 16 valves; (ii) the three SPE cartridges are fluidly connected to valves 18, 20, and 22; (iii) the six solvent vials are in fluid communication with the fourth, fifth, seventh, eighth, ninth, and tenth valves; (iv) the water vial is in fluid communication with a 15th valve; The method of claim 3.

5. The method of claim 4, wherein up to three vials of the composition are used.

6. In (ii) of claim 3, the cassette has a linear array of 25 valves; and (i) the 1 to 5 composition vials are fluidly connected to the 2, 12, 13, 14, and / or 16 valves; (ii) the two SPE cartridges are fluidly connected to the 20th and 22nd valves; (iii) the eight solvent vials are in fluid communication with the fourth, fifth, seventh, eighth, ninth, tenth, seventeenth, and eighteenth valves; (iv) the water vial is in fluid communication with a 15th valve; The method of claim 3.

7. The method of claim 6, wherein a maximum of two vials of the composition are used.

8. In (iii) of claim 3, the cassette has a linear array of 25 valves; and (i) the 1 to 5 composition vials are fluidly connected to the 2, 12, 13, 14, and / or 16 valves; (ii) the one SPE cartridge is in fluid communication with a 22nd valve; (iii) the ten solvent vials are in fluid communication with the fourth, fifth, seventh, eighth, ninth, tenth, seventeenth, eighteenth, nineteenth, and twentieth valves; (iv) the water vial is in fluid communication with a 15th valve; The method of claim 3.

9. The method of claim 8, wherein a maximum of one vial of the composition is used.

10. The cassette (i) a syringe in fluid communication with the third valve; (ii) a syringe in fluid communication with the eleventh valve; (iii) a syringe in fluid communication with the 24th valve; 10. The method of claim 4, further comprising:

11. The method of claim 10, wherein the syringe fluidly connected to the third valve is a 1 mL syringe.

12. The method of claim 10, wherein the syringe fluidly connected to the 11th valve is a 7 mL syringe.

13. The method of claim 10, wherein the syringe fluidly connected to the 24th valve is a 7 mL syringe.

14. 14. The method of claim 1, further comprising eluting impurities.

15. The method of claim 14, wherein the step of eluting the impurities is carried out before and / or after step (v).

16. 16. The method of any of claims 1 to 15, wherein the at least one solvent is selected from the group consisting of: (i) ethanol, (ii) methanol, (iii) acetonitrile, or a combination thereof.

17. The method of any preceding claim, further comprising conditioning the 1 to 3 SPE cartridges prior to step (ii).

18. 18. The method of any of claims 1 to 17, wherein said evaluating in step (vi) is carried out using high performance liquid chromatography (HPLC), liquid chromatography-mass spectrometry (LC-MS), or thin layer chromatography (TLC).

19. 19. The method of any of claims 1 to 18, wherein the elution is carried out by passing an organic solvent through the SPE cartridge and into the transfer line for collection.

20. The method described in claim 19, wherein the recovery is in a 96-well plate.

21. The 1 to 5 composition vials comprise: (i) 1 to 5 crude reaction mixture vials; (ii) 1 to 5 single reference standard vials; or (iii) 1 to 5 reference standard mixture vials 20. The method of any one of claims 1 to 19, wherein 22. The method of claim 21, wherein 1, 2, 3, 4 or 5 vials of the composition are used.

23. The method of claim 21 or 22, wherein one, two or three vials of the composition are used.

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