Apparatus for multiple simultaneous synthesis

• US 5,324,483 A
• Filed: 02/02/1993
• Issued: 06/28/1994
• Est. Priority Date: 10/08/1992
• Status: Expired due to Fees

• Current Assignee: Warner-Lambert Company (Pfizer Inc.)
• Original Assignee: Warner-Lambert Company (Pfizer Inc.)
• Inventors: Hodges, John C., Schroeder, Mel C., Moos, Walter H., Kiely, John S., Roth, Bruce D., Stankovic, Charles J., DeWitt, Sheila H. H., Pavia, Michael R., Cody, Donna R.
• Primary Examiner(s): Housel, James C.
• Assistant Examiner(s): EDWARDS, LAURA ESTELLE
• Application Number:
  US08/012,557
• Time in Patent Office: 511 Days
• Field of Search: 422/131, 422/99-104, 935/88, 530/333, 530/334, 211/71, 211/74, 435/300, 435/301
• US Class Current: 506/40
• CPC Class Codes: B01J 19/0046 Sequential or parallel reactions, e.g. for the synthesis of polypeptides or polynucleotides; Apparatus and devices for combinatorial chemistry or for making molecular arrays synthesis methods per se C40B50/00
• CPC Class Codes: B01J 2219/00283 Reactor vessels with top opening
• CPC Class Codes: B01J 2219/00308 interchangeably mounted in racks or blocks
• CPC Class Codes: B01J 2219/0031 the racks or blocks being mounted in stacked arrangements
• CPC Class Codes: B01J 2219/00373 Hollow needles
• CPC Class Codes: B01J 2219/00416 Vacuum
• CPC Class Codes: B01J 2219/00423 using filtration, e.g. through porous frits
• CPC Class Codes: B01J 2219/00454 by chemical cleavage from the solid support
• CPC Class Codes: B01J 2219/00493 by sparging or bubbling with gases
• CPC Class Codes: B01J 2219/00495 Means for heating or cooling the reaction vessels
• CPC Class Codes: B01J 2219/005 Beads
• CPC Class Codes: B01J 2219/00585 Parallel processes
• CPC Class Codes: B01J 2219/0059 Sequential processes
• CPC Class Codes: B01J 2219/00596 Solid-phase processes
• CPC Class Codes: B01J 2219/00689 using computers
• CPC Class Codes: B01J 2219/00695 Synthesis control routines, e.g. using computer programs
• CPC Class Codes: B01J 2219/0072 Organic compounds
• CPC Class Codes: B01J 2219/00722 Nucleotides
• CPC Class Codes: B01J 2219/00725 Peptides
• CPC Class Codes: C07K 1/045 using devices to improve synthesis, e.g. reactors, special vessels
• CPC Class Codes: C07K 1/047 : Simultaneous synthesis of different peptide species; Peptide libraries
• CPC Class Codes: C40B 40/06 : Libraries containing nucleotides or polynucleotides, or derivatives thereof
• CPC Class Codes: C40B 40/10 : Libraries containing peptides or polypeptides, or derivatives thereof
• CPC Class Codes: C40B 50/14 : Solid phase synthesis, i.e. wherein one or more library building blocks are bound to a solid support during library creation; Particular methods of cleavage from the solid support
• CPC Class Codes: C40B 60/14 : for creating libraries

Abstract

An apparatus and method which provides a suitable location for multiple, simultaneous synthesis of compounds. The apparatus consists of: a reservoir block having a plurality of wells; a plurality of reaction tubes, usually gas dispersion tubes, having filters on their lower ends; a holder block, having a plurality of apertures; and a manifold, which may have ports to allow introduction/maintenance of a controlled environment. The manifold top wall has apertures and a detachable plate with identical apertures. The apparatus is constructed from materials which will accommodate heating, cooling, agitation, or corrosive reagents. Gaskets are placed between the components. Rods or clamps are provided for fastening the components together. Apparatus operation involves placing the filters on the lower ends of the reaction tubes in the reservoir block wells, and the upper ends passing through the holder block apertures and into the manifold. The apparatus provides in excess of 1 mg of each product with structural knowledge and control over each compound. Using the apparatus a series of building blocks are covalently attached to a solid support. These building blocks are then modified by covalently adding additional different building blocks or chemically modifying some existing functionality until the penultimate structure is achieved. This is then cleaved from the solid support by another chemical reaction into the solution within the well yielding an array of newly synthesized individual compounds, which after postreaction modification, if necessary, are suitable for testing for activity.

Claims

1. An apparatus for multiple, simultaneous synthesis of compounds comprising:

a plurality of reaction tubes, each reaction tube having an upper end and a lower end, said lower end of each reaction tube comprising a filter, said upper end and said filter each having an inside diameter and an outside diameter;

a reservoir block having at least one reaction well for receiving said lower end of each reaction tube comprising the filter;

a holder block with a plurality of apertures, said apertures having a diameter at least equal to the outside diameter of at least one of said plurality of reaction tubes;

a manifold located adjacent to said holder block, said manifold enclosing the upper ends of said plurality of reaction tubes;

a means for providing a seal between said holder block and said reservoir block;

a means for providing a seal between said holder block and said manifold; and

a means for fastening together independently said reservoir block to said holder block and said holder block to said manifold.

2. An apparatus according to claim 1 in which the plurality of reaction tubes comprises at least one gas dispersion tube.

3. An apparatus according to claim 2, wherein said at least one gas dispersion tube has a length from 50 to 300 mm.

4. An apparatus according to claim 3 wherein the upper end of said at least one gas dispersion tube has an outside diameter from 5 to 25 mm.

5. An apparatus according to claim 3 wherein the upper end of said at least one gas dispersion tube has an inside diameter from 1 to 24 mm.

6. An apparatus according to claim 5 wherein the filter on the lower end of said at least one gas dispersion tube has a length from 1 to 300 mm.

7. An apparatus according to claim 6 wherein the filter on the lower end of said at least one gas dispersion tube has an outside diameter from 5 to 30 mm.

8. An apparatus according to claim 7 wherein the filter on the lower end of said at least one gas dispersion tube has an inside diameter from 1 to 24 mm.

9. An apparatus according to claim 8 in which the filter on the lower end of said at least one gas dispersion tube has a porosity of 10 to 400 μ

.

10. An apparatus according to claim 9 in which the filter on the lower end of said at least one gas dispersion tube is constructed of fritted glass.

11. An apparatus according to claim 10 wherein said at least one gas dispersion tube has a means for equilibrium of any pressure differentials within the sealed apparatus.

12. An apparatus according to claim 11 in which pressure equalization is achieved with a hollow tube placed around the upper end of said at least one gas dispersion tube, said hollow tube having one or more protrusions located in two or more locations along its inner face, such that said one or more protrusions contact the gas dispersion tube and prevent the gas dispersion tube from lateral or vertical movement.

13. An apparatus according to claim 11 in which pressure equalization is achieved with at least one capillary tube located adjacent and parallel to said at least one gas dispersion tube, said capillary tube extending from the reaction well, through an aperture in the holder block, into the manifold.

14. An apparatus according to claim 11 wherein the means for equilibrium comprises at least one pressure equalization hole located above the filter on the lower end of said at least one gas dispersion tube.

15. An apparatus according to claim 14 wherein the at least one pressure equalization hole is located from 1 to 250 mm above the filter on the lower end of the gas dispersion tube.

16. An apparatus according to claim 15 in which at least one pressure equalization hole has a diameter from 100 to 2000 μ

m.

17. An apparatus according to claim 1 in which the reservoir block comprises a reservoir rack which is adapted to hold at least as many reaction wells as the number of reaction tubes.

18. An apparatus according to claim 17 in which said at least one reaction well is sufficient in depth and diameter so that the reaction well may receive the filter on the lower end of one reaction tube and an amount of reactant required for organic synthesis.

19. An apparatus according to claim 18, wherein said at least one reaction well has an inside diameter of 7 to 100 mm.

20. An apparatus according to claim 19 in which said at least one reaction well is removable.

21. An apparatus according to claim 20 in which said at least one reaction well is constructed of glass.

22. An apparatus according to claim 1 in which at least one of the apertures in the holder block is coaxial to the apertures in the reservoir block.

23. An apparatus according to claim 22 in which the upper end of at least one of the plurality of reaction tubes passes through at least one of the plurality of apertures in the holder block.

24. An apparatus according to claim 1 in which the manifold has at least four side walls and one top wall, said top wall having an upper face and a lower face.

25. An apparatus according to claim 1 in which the manifold has a plurality of ports on one or more walls.

26. An apparatus according to claim 25 in which the manifold has two ports on one face.

27. An apparatus according to claim 26 in which the manifold is constructed of a transparent material.

28. An apparatus according to claim 27 in which the top wall of the manifold has a plurality of apertures which are coaxial to the plurality of apertures in the holder block.

29. An apparatus according to claim 28 in which a plate is located adjacent to the upper face of the top wall of the manifold.

30. An apparatus according to claim 29 in which the plate has a plurality of apertures which are coaxial to the plurality of apertures in the holder block.

31. An apparatus according to claim 1 in which the means for providing a seal between the holder block and the manifold comprises at least one first gasket.

32. An apparatus according to claim 1 in which the means for providing a seal between the holder block and the reservoir block comprises at least one second gasket.

33. An apparatus according to claim 30 in which the manifold and the plate are separated by at least one third gasket.

34. An apparatus according to claim 33 in which the at least one third gasket is penetrable with a needle object.

35. An apparatus according to claim 1 in which the reservoir block is releasably attached to the holder block and the manifold.

36. An apparatus according to claim 35 in which the means for fastening is one or more spring clips affixed to one or more sides of the reservoir block, the holder block, or the manifold.

37. An apparatus according to claim 36, in which the means for fastening is a rod which extends longitudinally upward throughout each of the reservoir block, the holder block, and the manifold.

38. An apparatus according to claim 32 in which a means for fastening the plate and the at least one-third gasket of the upper face of the manifold is two or more spring clips attached to at least one side wall of the manifold which snap into grooves on the upper face of the plate.

39. An apparatus according to claim 1 in which the means for providing a seal between the holder block, at least one reaction tube, and the manifold consists of an o-shaped ring.

40. An apparatus according to claim 39 in which a first spherical joint, opening downward and having a groove to accommodate the o-shaped ring, is fused to the outer wall of the reaction tube.

41. An apparatus according to claim 40 in which a second spherical joint having the same diameter and groove size of the first spherical joint and opening upward to accommodate the o-shaped ring is fused to the top of at least one reaction well.

42. An apparatus according to claim 41, wherein the first and second spherical joints have a diameter from 5 to 50 mm.

43. An apparatus according to claim 42 wherein the first spherical joint is located above the filter and the pressure equalization holes is at a height necessary to adequately accommodate the filter within a sealed reaction well when the seal is achieved.

44. An apparatus according to claim 43 in which the location of the first spherical joint is from 1 to 299 mm above the top of the filter.

45. An apparatus according to claim 44 in which the location of the second spherical joint fused to the top of the reaction well is also at a height necessary to adequately accommodate the filter within the sealed reaction well.

46. An apparatus according to claim 45 in which the location of the second spherical joint is from 1 to 299 mm above the bottom of the reaction well.

47. An apparatus according to claim 46 in which the face of at least one aperture on the lower side of the holder block is chamfered to accommodate at least one first spherical joint.

48. An apparatus according to claim 47 in which the face of at least one aperture on the upper side of the reservoir rack is chamfered to accommodate at least one second spherical joint.

49. An apparatus according to claim 1 in which a vacuum box having an open upper face surrounds the reservoir block.

50. An apparatus according to claim 49 in which a second rack which supports one or more removable cartridges is located adjacent to the open upper face of the vacuum box.

51. An apparatus according to claim 50 in which the means for providing a seal between the open upper face of the vacuum box and second rack is one or more gaskets adhered to the lower side of the base of the second rack.

52. An apparatus according to claim 51 in which the means for fastening the vacuum box and second rack is two or more spring clips attached to the side walls of the vacuum box and which snap into the grooves on the upper side of the base of the second rack.

53. An apparatus according to claim 52 in which one or more needle objects, corresponding in number and location to the reaction wells, are mounted in the base of the second rack, protrude through the gasket, and open into the test tubes.

54. An apparatus according to claim 53 in which the removable cartridges are used in combination with the needle objects.