APPARATUS AND METHOD FOR ITERATIVE POLYMER SYNTHESIS

Abstract

The present invention discloses a method and apparatus for fully automated iterative polymer synthesis at a large scale.

Claims

1-17. (canceled)

18. A method of iterative polymer synthesis comprising the following steps 1) to 11): 1) providing a plurality of movable, closable storage vessels (1), which are shaped so as to enable transport by an automated means (3) and each comprise a first transfer port (26) suitable for the transfer of solid material, wherein each of the storage vessels (1) contains a defined amount of a building block B in solid form to be used in one cycle of the iterative polymer synthesis process; 2) using an automated transport means (3) to transfer a specific storage vessel (1) selected from the plurality of y storage vessels (1) to a first reaction vessel RV1 (4), which comprises a second transfer port (27) suitable for the transfer of solid material, wherein: the first transfer port (26) of each storage vessel is constructed and arranged such that it can be releasably docked to the second transfer port (27) of the first reaction vessel RV1 (4), thereby forming a connection between both vessels; and material can pass though the transfer ports (26, 27) when in connected state, but not in disconnected state; 3) aligning and docking the first transfer port (26) on the storage vessel (1) to the second port (27) on the first reaction vessel RV1 (4); 4) opening the docked transfer ports (26, 27) and transferring the building block B from the storage vessel into the first reaction vessel RV1 (4); 5) dissolving the building block B by addition of a suitable solvent, thereby forming a solution of the building block B inside the first reaction vessel RV1 (4); 6) transferring the solution obtained in step 5), to a second reaction vessel RV2 (10) containing a carrier, to which a molecule C is tethered, thereby obtaining a reaction mixture containing the building block B.sub.i and the carrier with the molecule C; 7) cleaning the first reaction vessel RV1 (4), including the transfer port (27), by rinsing with solvent, which may be the same as or different from the solvent used in step 5); 8) incubating the reaction mixture obtained in step 6) under conditions, which allow for the formation of a chemical bond between the building block B.sub.i and the molecule C so as to form a molecule C′, which is extended by one building block unit; 9) retaining the carrier with the extended molecule C′ inside the second reaction vessel RV2 (10), while purging a liquid comprising byproducts and residual educts of the coupling reaction from the second reaction vessel RV2 (10); 10) conditioning the carrier with the extended molecule C′ for the next synthetic cycle with the extended molecule C′ as molecule C; and 11) undocking and removing the empty storage vessel from the first reaction vessel RV1(4) using the automated transport means (3); wherein at least the steps 2) to 11) are carried out at least once in automated fashion.

19. A method of iterative polymer synthesis comprising the following steps i) to xi): i) providing a plurality of movable, closable storage vessels (1), which are shaped so as to enable transport by an automated means (3) and each comprise a first transfer port (26) suitable for the transfer of solid material, wherein each of the storage vessels (1) contains a defined amount of a building block B in solid form to be used in one cycle of the iterative polymer synthesis process; ii) providing a first reaction vessel RV1 (4), which contains a carrier, to which a molecule C is tethered, and which comprises a second transfer port (27) suitable for the transfer of solid material, wherein: the first transfer port (26) of each storage vessel is constructed and arranged such that it can be releasably docked to the second transfer port (27) of the first reaction vessel RV1 (4), thereby forming a connection between both vessels; and material can pass though the transfer ports (26, 27) when in connected state, but not in disconnected state; iii) using an automated transport means (3) to transfer a specific storage vessel (1) selected from the plurality of storage vessels (1) to the first reaction vessel RV1 (4) containing the carrier with the molecule C; iv) aligning and docking the first transfer port (26) of the storage vessel (1) to the second transfer port (27) of the first reaction vessel RV1 (4); v) opening the docked transfer ports (26, 27) and transferring the amount of the building block B from the storage vessel into the first reaction vessel RV1 (4); vi) dissolving the building block B by addition of a suitable solvent, thereby forming a reaction mixture containing the building block B and the carrier with the molecule C inside the first reaction vessel RV1 (4); vii) incubating the reaction mixture obtained in step vi) under conditions, which allow for the formation of a chemical bond between the building block B and the molecule C so as to form a molecule C′, which is extended by one building block unit; viii) retaining the carrier with the extended molecule C′ inside the first reaction vessel RV1 (4), while purging a liquid comprising byproducts and residual educts of the coupling reaction from the first reaction vessel RV1 (4); ix) rinsing the first reaction vessel RV1 (4), including the second transfer port (27) with a solvent, which may be the same as or different from the solvent used in step vi); x) conditioning the carrier with the extended molecule C′ for the next coupling round with the extended molecule C′ as molecule C; and xi) undocking and removing the empty storage vessel from the first reaction vessel RV1(4) using the automated transport means (3); wherein at least the steps iii) to xi) are carried out at least once in automated fashion.

20. The method according to claim 18, wherein the first transfer port (26) comprises the passive part (2) of a split valve device, which is suitable for the transfer of solid material, and the second transfer port (27) comprises the active part (5) of the split valve device.

21. The method according to claim 18, wherein the plurality of movable, closable storage vessels (1) comprises at least y of the movable, closable storage vessels (1), wherein each of the y storage vessels (1) contains a defined amount of a building block B.sub.i in solid form to be used in one cycle of the iterative polymer synthesis process, where y is an integer equal to or larger than 2 and i is an index ranging from 1 to y, and wherein y synthetic cycles are carried out, each comprising the steps 2) to 11).

22. The method according to claim 18, further comprising a step of adding an activating reagent to a solution of step 5) in at least one synthetic cycle.

23. The method according to claim 18, further comprising a step of using a device M to determine the amount of building block B, which has been transferred from the storage vessel (1) into the reaction vessel first reaction vessel RV1 (4).

24. An apparatus suitable for performing iterative polymer synthesis, comprising: a) a plurality of movable, closable storage vessels (1), which are shaped so as to enable transport by an automated means (3) and each comprise a first transfer port (26) suitable for the transfer of solid material; b) at least one first reaction vessel RV1 (4), which comprises a second transfer port (27) suitable for the transfer of solid material, wherein: the first transfer port (26) of each storage vessel is constructed and arranged such that it can be releasably docked to the second transfer port (27) of the first reaction vessel RV1 (4), thereby forming a connection between both vessels; and material can pass though said transfer ports (26, 27) when in connected state, but not in disconnected state; c) an automated transport means (3) suitable for bringing a defined sequence of individual storage vessels (1) to a specific first reaction vessel RV1 (4) and away from it, wherein the automated transport means is capable of aligning the transfer port (26) on the storage vessel (1) with the transfer port (27) on the first reaction vessel RV1 (4) with sufficient precision to enable their docking to each other; and d) at least one control unit CU1(7) controlling the actions of the automated transport means (3), the docking of the ports (26, 27), and the opening and closing of the port, wherein the apparatus is configured to carry out the method according to claim 21.

25. The apparatus according to claim 24, wherein the first transfer port (26) comprises the passive part (2) of a split valve device, which is suitable for the transfer of solid material, and the second transfer port (27) comprises the active part (5) of the split valve device.

26. The apparatus according to claim 24, wherein the automated transport means comprises a robot or a conveyor device.

27. The apparatus according to claim 24, wherein the automated transport means comprises a robotic arm equipped with a gripping device (24).

28. The apparatus according to claim 24, wherein each of the storage vessels (1) comprises a liquid inlet (8), which allows connection to a mobile solvent line (9).

29. The apparatus according to claim 24, wherein the first reaction vessel RV1 (4) comprises a liquid inlet (8), which allows connection to a liquid line (11).

30. The apparatus according to claim 24, further comprising a device M (18) for monitoring the extent of material transfer from the storage vessel to the first reaction vessel RV1 (4).

31. The apparatus according to claim 24, further comprising at least one cleaning device (17) for the active part (5) of the split valve device of the first reaction vessel(s) RV1 (4), wherein the actions of the cleaning device are controlled by the control unit CU1 (7).

32. The apparatus according to claim 24, further comprising: e) at least one second reaction vessel RV2 (10) connected to at least one of the one or more first reaction vessel(s) RV1 (4) f) a device (20) for controlling liquid flow from the first reaction vessel(s) RV1 (4) into the connected second reaction vessel(s) RV2 (10), which is controlled by the control unit CU1 (7).

33. The apparatus according to claim 31, further comprising a waste line (13) allowing to drain liquid from the first reaction vessel RV1 (4) and from a liquid connection line (6) connecting the first reaction vessel RV1 (4) with the second reaction vessel RV2 (10), without passage of the liquid through the second reaction vessel RV2 (10).

34. The apparatus according to claim 32, comprising a number of n first reaction vessels RV1 (4) and a number of m second reaction vessels RV2 (10), wherein n and m are integers chosen independently from the range of 1 to 10.

35. The apparatus according to claim 24 comprising at least one reaction vessel RV1 (4) or RV2 (10), wherein the reaction vessel or reaction vessels further comprise(s) one or more elements selected independently for each reaction vessel from the group consisting of a sensor (16), a heating and/or cooling device (19), a mixing device (14), a liquid line (11), a liquid port (29), a means (28) for rinsing the inner walls of the reaction vessel, and a means (15) for separating the carrier with the growing polymer chain from the remaining components of the reaction mixture.

36. The method according to claim 19, wherein the first transfer port (26) comprises the passive part (2) of a split valve device, which is suitable for the transfer of solid material, and the second transfer port (27) comprises the active part (5) of the split valve device.

37. An apparatus suitable for performing iterative polymer synthesis, comprising: g) a plurality of movable, closable storage vessels (1), which are shaped so as to enable transport by an automated means (3) and each comprise a first transfer port (26) suitable for the transfer of solid material; h) at least one first reaction vessel RV1 (4), which comprises a second transfer port (27) suitable for the transfer of solid material, wherein: the first transfer port (26) of each storage vessel is constructed and arranged such that it can be releasably docked to the second transfer port (27) of the first reaction vessel RV1 (4), thereby forming a connection between both vessels; and material can pass though the transfer ports (26, 27) when in connected state, but not in disconnected state; i) an automated transport means (3) suitable for bringing a defined sequence of individual storage vessels (1) to a specific first reaction vessel RV1 (4) and away from it, wherein the automated transport means is capable of aligning the transfer port (26) on the storage vessel (1) with the transfer port (27) on the first reaction vessel RV1 (4) with sufficient precision to enable their docking to each other; and j) at least one control unit CU1(7) controlling the actions of the automated transport means (3), the docking of the ports (26, 27), and the opening and closing of the port, wherein the apparatus is configured to carry out the method according to claim 22.

38. The method according to claim 19, wherein the plurality of movable, closable storage vessels (1) comprises at least y of the movable, closable storage vessels (1), wherein each of the y storage vessels (1) contains a defined amount of a building block B.sub.i in solid form to be used in one cycle of the iterative polymer synthesis process, where y is an integer equal to or larger than 2 and i is an index ranging from 1 to y, and wherein y synthetic cycles are carried out, each comprising the steps iii) to xi).

39. The method according to claim 19, further comprising a step of adding an activating reagent to a solution of step vi) in at least one synthetic cycle.

Description

EXAMPLES

Example 1: Flowability and Solubility of Amino Acid Derivatives

[0289] The flowability of 15 samples of amino acid powders was tested. The flowability of each powder in unconfined state was assessed by powder flow analysis in a rotating drum (“revolution analyzer”), by testing mass flow through a funnel, and by determining the angle of repose. The latter two measurements were performed according to chapter 1174 of the U.S. Pharmacopeia. The flowability of the samples at a confided state was analyzed by measurements with an annular shear cell and by using the Evolution Powder Tester device by PS Prozesstechnik GmbH, Basel. In these tests, the samples were initially densified with different pre-consolidation forces.

[0290] In the tests without confinement, Fmoc-Cys(Trt)-OH and Fmoc-Met-OH consistently stood out for good flowability, and flowability parameters for Fmoc-Gly-OH were likewise good. On the other hand, poor flowability parameters were determined for Fmoc-Ala-OH and Fmoc-Pro-OH. Interestingly, in the tests with confinement, Fmoc-Cys(Trt)-OH and Fmoc-Pro-OH consistently stood out for good flowability. Medium values were determined for Fmoc-Ala-OH, while Fmoc-Gly-OH and Fmoc-Lys(Boc)-OH consistently stood out for poor flowability. This might indicate that Fmoc-Gly-OH may lose its flowability upon compression. On the other hand, the compression may have broken down lumps in the Fmoc-Pro-OH sample, thereby increasing its flowability.

[0291] The concentration of the same amino acids in saturated solutions of dimethyl formamide at room temperature was determined by analytical reversed phase ultrahigh pressure liquid chromatography. The values ranged from below 200 mg/ml for Fmoc-Pro-OH over around 400 mg/ml for Fmoc-Ala-OH to above 600 mg/ml for Fmoc-Gly-OH.

Example 2: Transfer of an Amino Acid Derivative Powder Through a Split Valve Device

[0292] Fmoc-Ala-OH and Fmoc-Gly-OH were chosen as test substances because of their poor flowability revealed by the previous experiments.

[0293] 2 kg of amino acid derivative powder were filled into a 25 l storage vessel made of stainless steel. The storage vessel was closed by the passive part of a DN100 split valve device, i.e. a split valve device with an inner opening of roughly 10 cm. The active part of the split valve device was docked and a transparent bag attached to it. The split valve device was opened manually and the flow of the powder observed by eye though the transparent bag. A pulsating vibrator was applied to the active part of the split valve device to facilitate the powder flow. The powder transfer was completed within a maximum time of 5 minutes. Visual inspection was performed by opening the valve. Remnants of white powder were observed on the walls of the vessel and valve an on the valve's flap.

[0294] The vessel was subsequently rinsed with ca. twenty liters of dimethyl formamide though a fixed spray ball, which was placed inside the storage vessel opposite to the split valve device and was connected to a solvent line. Visual inspection was repeated after rinsing. No remnants were observable.

[0295] These findings demonstrate that the powder transfer is achievable within a suitable time frame and that efficient rinsing of the split valve device is possible. They therefore can be considered a proof of concept for the present invention.

LIST OF REFERENCE SIGNS

[0296] 1 storage vessel [0297] 2 passive part of split valve device [0298] 3 automated transport means [0299] 4 first reaction vessel RV1 [0300] 5 active part of split valve device [0301] 6 liquid connection line [0302] 7 control unit CU1 [0303] 8 liquid inlet [0304] 9 mobile solvent line [0305] 10 second reaction vessel RV2 [0306] 11 liquid line [0307] 12 drain [0308] 13 waste line [0309] 14 mixing device [0310] 15 separation means [0311] 16 sensor [0312] 17 cleaning device [0313] 18 device M for monitoring material transfer [0314] 19 heating/cooling device [0315] 20 device controlling liquid flow [0316] 21 connection to source of protective [0317] 25 gas [0318] 22 connection to vacuum source [0319] 23 pressure controller [0320] 24 gripping device [0321] 25 gripper plate [0322] 26 first solid material transfer port [0323] 27 second solid material transfer port [0324] 28 rinsing means [0325] 29 liquid port

BRIEF DESCRIPTION OF THE FIGURES

[0326] FIG. 1 shows a basic layout of some embodiments according to the present invention. Out of a plurality of such storage vessels, an automated transport means (3) selects a storage vessel (1) comprising a passive part (2) of a split valve device. The automated transport means (3) transfers said storage vessel (1) to a first reaction vessel RV1 (4), and aligns the passive part (2) of the split valve device with an active part (5) of said split valve device on the first reaction vessel RV1. The active and passive parts (2, 5) of the split valve device are docked and the valve opened to allow for passage of solid material from the storage vessel (1) into the first reaction vessel RV1 (4). After the transfer, the split valve device is closed and undocked and the storage vessel (1) is transported away from the first reaction vessel RV1 (4). Liquid may be added to the first reaction vessel RV1 (4) via a liquid line (11) and drained via a line (6). A device controlling liquid flow (20) is integrated into the liquid line (6). The device for controlling liquid flow (20) is, for example, constructed as a valve. The actions of the automated transport means (3), the docking/undocking of the split valve device, and the opening/closing of the split valve device are controlled by a control unit CU1 (7).

[0327] FIG. 2 shows various embodiments of the automated transport means (3). a) the automated transport means (3) comprises a conveyer system, where individual storage vessels (1) are transported along rails into the docking position on top of the first reaction vessel RV1 (4) and back. b) the automated transport means (3) comprises a “carousel type” conveyer system, where individual storage vessels (1) are transported by rotation of a wheel into the docking position on top of the first reaction vessel RV1 (4) and back. c) the automated transport means (3) comprises a robotic arm, which engages with an individual storage vessel (1) via a gripping device (24), and shifts it into the docking position on top of the first reaction vessel RV1 (4) and back.

[0328] FIG. 3 shows one embodiment of the apparatus according to the present invention comprising one first reaction vessel RV1 (4) connected to a second reaction vessel RV2 (10) via a liquid connection line (6). A device controlling liquid flow (20) is integrated into the liquid connection line (6). The storage vessel (1) comprising a passive part (2) of a split valve device, the automated transport means (3) and the first reaction vessel RV1 (4) with the active part (5) of said split valve device are as described with respect to FIG. 1. A cleaning device (17) allows to clean the active part (5) of the split valve device on the first reaction vessel RV1 (4). The first reaction vessel RV1 (4) and the second reaction vessel RV2 (10) are each equipped with a connection (21) to a source of protecting gas, with a pressure controller (23), and with a connection (22) to a vacuum source. Various liquid lines (11) allow for addition of solvent and liquid reagents. Liquid flow from the first reaction vessel (4) to the second reaction vessel RV2 (10) may be driven by any suitable means, e.g. by an overpressure applied to first reaction vessel RV1 (4), by gravity, by a vacuum applied to second reaction vessel RV2 (10), or by a pump [not pictured]. Liquid may be drained from the second reaction vessel RV2 (10) via a drain (12), in which a device controlling liquid flow (20) is integrated. A control unit CU1 (7) controls at least the actions of the actions of the automated transport means (3), the docking/undocking of the split valve device, the opening/closing of the split valve device, the devices controlling liquid flow (20), and the cleaning device (17). The control unit CU1 (7) may further control the dosing of solvent and reagents via the liquid lines (11), the settings of the pressure controller (23), the flow of protective gas into the reaction vessels RV1 and RV2, and/or the vacuum suction applied to the reaction vessels RV1 and RV2.

[0329] FIG. 4 shows one embodiment of the apparatus according to the present invention comprising three separate synthesis lines, each comprising one first reaction vessel RV1 (4) connected to a second reaction vessel RV2 (10). A single automated transport means (3) allows to bring a selected storage vessel (1) to a selected first reaction vessel RV1 for docking and material transfer. The other elements of the figure are as set out with respect to FIG. 3. This embodiment of the apparatus thus allows to carry out three independent and different synthesis reactions in parallel.

[0330] FIG. 5 shows another embodiment of the apparatus according to the present invention. Each of the storage vessels (1) comprises a liquid inlet (8) feeding into a rinsing means (28) inside the storage vessel (1). A mobile solvent line (9) may be connected to the liquid inlet (8) of any storage vessel (1). Further, liquid lines (11) are each connected to a liquid inlet (8) feeding into a rinsing means inside the first reaction vessel RV1 (4) and inside the second reaction vessel RV2 (10), respectively. An additional liquid port (29) is provided on the first reaction vessel RV1 (4). This may allow for connection to incoming or outgoing lines. Both reaction vessels (4, 10) further comprise a mixing device (14),—e.g. a stirrer—, a sensor (16)—e.g. a temperature sensor, a pressure sensor, a level sensor, a turbidity sensor, an optical sensor, a conductivity sensor, an impedance sensor-, and a heating/cooling device (19); the second reaction vessel RV2 (10) additionally comprises a separation means (15) installed in the lower part of the vessel. This separation means (15) may allow to separate the carrier with the growing polymer chain from the remaining components of the reaction mixture by retaining the carrier inside the second reaction vessel RV2 (10) while draining the majority of other components via the drain (12). The cleaning device (17), which in this embodiment comprises a passive part (2) of the split valve device, allows to clean the active part (5) of the split valve device on the first reaction vessel RV1 (4). The liquid used for the cleaning process may be drained via a waste line (13). A monitoring device (18)—e.g. a scale—allows to verify whether material transfer from the storage vessel (1) to the first reaction vessel RV1 (4) is sufficiently complete. The other elements of the figure are as set out with respect to FIG. 3.

[0331] FIG. 6 shows one embodiment of the apparatus according to the present invention comprising two first reaction vessels RV1 (4), each connected to two second reaction vessels RV2 (10). A single automated transport means (3) allows to bring a selected storage vessel (1) to a selected first reaction vessel RV1 (4) for docking and material transfer. The other elements of the figure are as set out with respect to FIG. 3. This embodiment of the apparatus thus allows to carry out two independent and different synthesis reactions in parallel. In addition, two first reaction vessels RV1 (4) may be used in parallel for preparing building block solutions to be added to the same second reaction vessel RV2 (10).

[0332] FIG. 7 shows another embodiment of the apparatus according to the present invention, which comprises a plurality of storage vessels (1), each comprising a first solid material transfer port (26) and an optional gripper plate (25). An automated transport means (3) selects a storage vessel (1), transfers said storage vessel (1) to a first reaction vessel RV1 (4), and aligns the first solid material transfer port (26) of the storage vessel with a second solid material transfer port (27) on the first reaction vessel RV1 (4). The solid material transfer ports (26, 27) are docked together and subsequently opened to allow for passage of solid material from the storage vessel (1) into the first reaction vessel RV1 (4). After the transfer, the solid material transfer ports (26, 27) are closed, the vessels undocked, and the storage vessel (1) is transported away from the first reaction vessel RV1 (4). Liquid may be drained from the first reaction vessel RV1 (4) via a liquid connection line (6). A device controlling liquid flow (20) is integrated into the liquid connection line (6). The actions of the automated transport means (3), the docking/undocking of the solid material transfer ports (26, 27), and the opening/closing of the solid material transfer ports (26, 27) are controlled by a control unit CU1 (7). In the embodiment pictured, the first reaction vessel RV1 (4) further comprises an optional mixing device (14) and an optional separation means (15) installed in the lower part of the vessel.