Note: Descriptions are shown in the official language in which they were submitted.
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WALDEMAR RUEDIGER 8MS-29
WEN-JENG LI
JOHId W . ALLEN, JR .
HAROLD N. WELLER, III
APPARATUS POR
SYNTHESIS OP MULTIPLE ORGANIC COMPOUNDS
WITH PINCH VALVE $LOCR
The present invention relates to apparatus for combina-
torial drug research to be used in the simultaneous parallel solid
and solution phase synthesis of large numbers of diverse organic
compounds or for the final cleavage step of radio frequency tagged
synthesis and more particularly to a modular apparatus designed for
such purposes which employs a unique pinch valve block, which
includes reactor vessels capable of receiving porus polyethelyene
microcannisters with radio frequency transmitter tags and which can
be used to discharge into all of the wells of a standard microtiter
plate.
Efficient testing of organic compounds in the modern
pharmaceutical laboratory requires the synthesis of large numbers
of diverse organic molecules in an automated and high speed manner.
The apparatus of the present invention is designed for use in such
a system, particularly one which employs solid phase synthesis
techniques. It is useful in performing the entire synthesis or for
performing only the final cleavage step of radio frequency tagged
synthesis.
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During the course of the synthesis, various operations
must be performed on the samples, including reagent introduction
and removal, agitation, washing, and compound removal by cleavage
from a resin support. Precise control of temperature, pressure and
atmospheric gas mixtures may be required at various stages. These
operations are standard and can be performed at task specific work
stations which have been designed or modified for use with one or
more reactors.
Over the last few years, a number of different systems
have been developed to produce libraries of large numbers of speci-
fic types of organic molecules, such as polynucleotides. However,
the usefullness of such systems tends to be limited to the particu-
lar type of molecule the system was designed to produce. Our
invention is much more general in application. It can be used to
synthesize all types of organic compounds including those used in
pharmaceutical research, the study of DNA, protein chemistry,
immunology, pharmacology or biotechnology.
Aside from the lack of versatility, existing equipment
for automated organic synthesis tends to be large and heavy, as
well as very expensive to fabricate and operate. Known automated
systems also tend to be quite complex, requiring equipment which is
limited as to flexibility, speed, and the number and amount of com-
pounds which can as be synthesized. As will become apparent, our
system has a simple, elegant design. It is relatively inexpensive
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to fabricate and operate. However, it is extremely flexible and is
capable of producing large numbers and amounts of all types of
organic cocapounds in a high. speed, automated manner. It is smaller
in size than comparable equipment, permitting more reactors to be
used at one time at a work station and it is lighter, thereby
facilitating movement of the apparatus between work stations with
less effort.
One system of which we are aware was developed for use at
Zeneca Pharmaceuticals, Alderley Park, Macclesfield, Cheshire, SK10
4TG, United ~ Kingdom. That system is built around an XP Zymate
laboratory robot (Zymark Corporation, Hopkinton, MA). The robot
arm is situated in the middle of a plurality~of stationary work
stations arranged in a circle. The arm is programmed to move one
or more tube racks from one station to another. However, the
Zeneca system has a small throughput capability, as the number of
tube racks which can be handled at one time is limited.
An automated peptide synthesizer developed for Chiron
Corporation of Emeryville, CA., which has similar limitations, is
described by Ronald N. Zukermann, Janice_M. Kerr, Michael A. Siani
and Steven C. Banville in an article which appeared in the Interna-
tional Journal of Peptide and Protein Research, Vol. 40, 1992,
pages 497-506 entitled "design, Construction and Application of a
Fully Automated Equimolar Peptide Mixture Synthesizer". See also
U.S. Patent No. 5,240,680 issued August 31, 1993 to Zuckermann and
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Banville and U.S. Patent No. 5,252,296 issued October 12, 1993 to
Zukertnann et al. entitled "Method and Apparatus For Biapolymer
Synthesis".
Another approach was developed at Takeda Chemical Indus-
tries, Ltd. and is described in an article published in the Journal
of Automatic Chemistry, Vol. 11, No. 5 (Sept.-Oct. 1989) pp. 212-
220 by Nobuyoshi Hayashi, Tobru Sugawara, Motoaki Shintani and
Shinji Kato entitled "Computer-assisted Automatic Synthesis II.
Development of a Fully Automated Apparatus for Preparing Substi-
tuted N- (carboxyalkyl) Aminio Acids". The Takeda system includes
a plurality of stationazy units.which are computer controlled. The
reactor unit includes only two reaction flasks. A plurality of
computer controlled solenoid valves regulate the input flow from
the reactant supply unit and wash solvent supply unit as well as
output to the purification unit, exhaust and drainage unit. Sen-
sors and electrodes feed information back to the computer. That
system is complex, costly and inflexible. It is also very limited
with respect to the number of compounds which can be synthesized.
A more flexible approach has been suggested by the Parke-
Davis Pharmaceutical Research Division of Warner-Lambert, as des-
cribed by Sheila Hobbs DeWitt et al. in Proc. National Academy of
Science, USA, Vol. 90, pp. 6909-6913 August 1993 and in the ISLAR
'93 Proceedings. That system employs a Tecan robotic sample pro-
cessor. A manifold of gas dispersion tubes are employed in combi-
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nation with glass vials. The glass frits of the tubes contain the
solid support during reactions. However, like many prior art
systems, in this apparatus, samples from the reaction tubes must be
removed from above, using a modified needle as a probe. There is
no facility for removal from the bottoms of the tubes. Accord-
ingly, obtaining product from the reactor vessels in the Parke-
Davis system is awkward and time consuming.
U.S. Patent No. 5,472,672 issued December 5, 1995 to
Thomas Brennan, entitled "A,pparatus and Method for Polymer Synthe-
sis Using Arrays", teaches the use of an automated system in which
a~ transport mechanism is used to move a base having an array of
reactor wells in conveyor belt fashion from work station to work
astation. Sample removal is performed by creating a pressure dif-
ferential between the ends of the wells. Aside from the difficul-
ties with regard to discharge, this system is complex and lacks
flexibility.
H'e are also aware of system designed by the Ontogen
Corporation of Carlsbad, CA 92009 as disclosed by John Cargill and
Romaine Maiefski in Laboratory Robotics and Automation, Vol. 6 pp.
:139-147 in an article entitled "Automated Combinatorial Chemistry
on Solid Phase" and disclosed in U.S. Patent No. 5, 609, 826 entitled
"Methods and Apparatus for the Generation of Chemical Libraries"
issued March 11, 1997 to John Cargill and Romaine Maiefski. The
system disclosed in the article and patent utilizes a reactor block
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having an array of reactor vessels. The block is moved along an
assembly line of work stations under computer control.
The Ontogen apparatus disclosed in the above mentioned
article and patent has a number of shortcomings. It is highly
complex and expensive. 7:t does not include any valuing structure
capable of regulating the fluid discharge from the reactor
chambers. Instead, it depends upon pressure differential to cause
discharge through s-shaped trap tubes which snap into a fitting on
the bottom of each react:Lon vessel. This takes up a lot of room,
preventing the dense packing of the reactor vessels. It also makes
product removal~awkward.
Because the reactor vessels disclosed in the article and
patent cannot be densely;packed, mirror image reactors are required
in the Ontogen system to discharge into all of the densely packed
wells of a standard microtiter plate. As described in U.S. Patent
No. 5,609,826, two different reactor configurations, each capable
of receiving a set of 48 reaction vessels, are required to deposit
directly into all 96 of the microtiter wells.
Reactor vessels of the type commonly used in the art are
not adapted to receive commercially available pores polyethelene
microcannisters. As is disclosed in the literature noted below,
such microcannisters can be radio frequency transmitter tagged for
automated tracking. Hence, it would be very advantageous to have
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a reactor which could deposit into all the microtiter wells and
still utilize reactor vessels capable of receiving commercially
available microcannisters.
International Publication Number WO 97/10896 under the
Patent Cooperation Treaty published on March 27, 1997 teaches
apparatus for simultaneous solid phase chemical synthesis developed
by Berlex Laboratories, Ine, of Richmond, California. The Berlex
equipment utilizes a manifold valve block including a plurality of
aligned valve inserts which are controlled by valve stems. The
stems are rotated by hydraulic cylinders positioned on either side
of the manifold. The Berlex apparatus accomodates 96 reactor
vessels at one time in a densely packed array. however, the
reactor vessels cannot receive porous microcannisters with radio
frequency tags. Moreover, this reactor requires a specially
designed solvent delivery system.
Personnel at Bristol-Myers Squibb Company of Princeton,
New Jersey 08543 developed an earlier version of the present
apparatus designed for use in the simultaneous synthesis of diverse
organic compounds. Like the present invention, it consisted of
stackable modules which are moveable among nesting sites located on
work station platforms. The reactor module in that version in-
cludes a heat transfer block adapted to receive an array of reactor
vessels. The reactor vessels are in the form of solid phase ex-
traction cartridges without sorbent. Each has a bottom outlet
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port. A plurality of separate valves arranged in rows are located
below the vessels. The valves consist of stopcocks which are gang-
controlled to regulate the discharge from the reactor vessel outlet
ports into aligned channels, each formed by a pair of threated Leur
tip adapters. The reactor module is situated over a discharge
module. The inlet openings in the discharge module are adapted to
accept the threaded ends of the Leur tip adapters. The discharge
module consists of a multi-well collector block or a drain block.
A solvent introduction module, which includes a pressure plate hav-
ing an array of openings and a septum, is received over the reactor
module. The downwardly projecting rim defining each pressure plate
opening cooperates with the septum to engaged the mouth of the
aligned reactor vessel t.o maintain a fluid tight seal.
Although that apparatus was a vast improvement over the
prior art systems, it stall had some disadvantages. For example,
the apparatus was still relatively large and has connectors and
levers extending outwardly from the sides, allowing only two
reactors to fit under a standard fume hood at one time. Each
reactor weighed about 18 pounds and was costly to fabricate. Thus,
improvement in the areas of size, weight and cost are possible. A
more elegant valve system, with fewer moving parts, is also desire-
able. Provision for receiving commercially available porous micro-
cannisters with radio frequency transmitter tags for automated
encoding in the reactor- vessels would be extremely advantagous.
Moreover, a structure which could accommodate standard microtiter
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plates or blocks for specimen collection would be an important
advance. Improvements i.n these areas are embodied in the present
invention.
Our approach to the automation problem in this invention
is to employ modules of simplified design and construction which
can be readily arranged in sets to perform the required operations
and which are light in weight so as to be easily moveable among
nest sites at standard work stations. This permits the greatest
amount of flexibility ait the least cost. . Due to more compact
design, more reactors can be assembled and employed at one time by
creating multiple nest sites at a single work station, such as an
orbital shaker. For time consuming operations, several work
stations can be in use simultaneously, to permit parallel flow of
reactors and therefore eliminate bottlenecks. For less time con-
suming operations, fewer work stations can be used, as long as the
flow of reactors is not impeded. Because the reactors are lighter
in weight, they are easier to transport. Accordingly, maximum
throughput is acheived with minimum investment.
In addition, the apparatus of the present invention is
designed to permit sample removal from the bottom of the reactor
vessels as in the earlier version of the Bristol-Myers Squibb
equipment. However, unlike the earlier equipment system, the
present invention employes simplified valuing in the form of a
unique pinch valve block located beneath the reactor block. The
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valve block includes plates with sets of aligned, relatively move-
able ribs . Each rib set is aligned with the outlet tubes asso-
ciated with a different row of reactor vessels. Movement of the
rib surfaces causes force to be applied to the outlet tubes through
Telfon encapsulated silicone 0-ring cord sections situated between
one rib surface and the .adjacent outlet tubes, such that the tubes
are simultaneously closed (pinched) without crushing or damaging
the tube walls. As a result, the tube walls will reliably resume
their original open condition each time the force is released.
The apparatus of the present invention includes a reactor
block, located above the valve block, which accepts an array of
reactor vessels . The vessels may be any plastic or glass tube with
a bottom port, such as a standard solid phase extraction cartidge
without sorbent. However, the reactor vessels are preferrably
designed to receive poi-us polyethelene microcannisters provided
with radio frequency transmitter tags for automated tracking. The
apparatus can be used for the entire synthesis or only the final
cleavage step in radio frequency tagged synthesis, as desired.
The reactor module is adapted to mount over a discharge
module. The discharge module may consist of a collection block
with an array of wells for collection tubes or vessels. Prefer-
rably, it takes the forth of a 96 well microtiter block of standard
size and dimension. If the reactor vessels are large enough to
accept cotttmercially available porus microcannisters, they may be
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too large to permit them to.be packed tightly enough to discharge
into all of the 96 wells of a standard microtiter block at once.
A funnelling device could be interposed between the valve block and
the microtiter plate to direct the discharge from the reactor
vessels into the wells of the plate. However, such a device is
bulky and expensive to fabricate. Alternatively, as in the Ontogen
system mentioned above and disclosed in U.S. Patent NO. 5,609,826,
mirror image reactors (referred to as type "A" and type "B") could
employed, each capable of holding 48 reactor vessels and discharg-
ing into a different set of 48 wells in the 96 well microtiter
plate.
Our system overcomes the costs and problems of requiring
an interposed funnelling device or having two reactor configura-
tions by employing a single reactor block with 52 possible reactor
vessel positions, instead of the conventional 48. Either one of
two different sets (referred to as "odd" or "even") of 48 positions
out of the possible 52 positions can be selected for use. By
shifting the position of the reactor block relative to the micro-
titer plate, discharge into either the odd or even well set in the
microtiter plate can be achieved.
Internal vertical supports are employed to facilitate
alignment of the blocks as the reactor module sets are formed. The
supports each have a plurality of'different levels. Different
blocks are designed to rest on different levels. In this way, dif-
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ferent reactor configurations are easily formed. For example,
reactors with or without temperature control blocks can be assembl-
ed. Simple nesting brackets with chamfered surfaces make installa-
lion of the reactors on the work stations a quick and easy task.
Since the modifications to standard work stations to
accept the reactors of the present invention are simple and in-
expensive to make, little time or cost is involved in converting a
conventional laboratory for use with the system of present inven-
lion. This dramatically increases the speed of the set up of a
facility to perform ~the synthesis process, as customized work
stations, specialized computers and complex interfaces are not
required.
It is, therefore, a prime object of the present invention
to provide apparatus for the synthesis of multiple organic com-
pounds which is mechanically simple, small in size, light in
weight, relatively inexpensive to construct, does not require ex-
tensive set up time, is extremely flexible and has high throughput.
It is another object of the present invention to provide
apparatus for the synthesis of multiple organic compounds which
consists of a plurality of stackable modules adapted to be moved as
a unit among nest sites on work station platforms.
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It is another object of the present invention to provide
apparatus for the synthesis of multiple organic compounds which can
be used for performing the entire synthesis or only the final
cleavage step of radio frequency tagged synthesis.
It is another object of the present invention to provide
apparatus for the synthesis of multiple organic compounds which
includes a multiple valve block in which sets of aligned ribs of
relatively moveable plata_s act through Teflon encapsulated silicone
O-ring cord sections to close rows of outlet tubes to regulate the
discharge from the reactor vessel ports.
It is another object of the present invention to provide
apparatus for the synthesis of multiple organic compounds which is
compatible for use with a standard 96 well microtiter collection
plate where a single configuration reactor block with 52 reactor
vessel positions can be employed to discharge into either the even
or the "odd" 48 well sets of the plate by shifting the relative
position of the microtiter plate.
It is another object of the present invention to provide
apparatus for the synthesis of multiple organic compounds which
utilizes reactor vessels adapted to receive porus microcannisters
with radio frequency transmitter tags.
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In accordance with one aspect of the present invention,
apparatus useful for the synthesis of multiple organic compounds is
provided. The apparatus is adapted to receive an array of indivi-
dual reactor vessels. Each vessel has a port connected to an out-
let tube. Valve means simultaneously regulate the discharge from
the vessels through the outlet tubes. The valve means includes
first and second relatively moveable surfaces between which the
outlet tubes extend. Resilient means are interposed between one of
the surfaces and the outlet tubes. Relative movement of the sur-
faces causes force to be applied through the resilient means to
close the outlet tubes.
The valve surfaces are the surfaces of ribs located on
relatively moveable plates. The resilient means preferrably takes
the form of Teflon encapsulated silicone O-ring cord, cut in
sections and situated adjacent one of the rib surfaces. The rib
surface adjacent the cord is shaped to correspond to the shape of
the cord. More specifically, it has an arcuate shape which serves
to maintain the cord secaion in proper position. The ends of one
of the plates have openings through which the resilient means can
be inserted so as to be received between the ribs.
The valve means is located below the reactor vessels.
Below the valve means is situated the collection block. The
collection block has an array of wells. A plurality of collection
vessels are adapted to be received in the wells.
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The collection block is capable of receiving 2X number of
collection vessels. The apparatus is adapted to receive X number
of reactor vessels in Y number of possible positions, where Y is a
number larger than X. The collection block can be received in one
of two positions relative to the reactor block.
The reactor vessels may receive porus polyethylene micro-
cannisters with radio frequency transmitter tags. Multi-level com-
ponent internal support and alignment means are provided.
In accordance with another object of the present inven-
tion, valve means are provided for the simultaneous regulation of
the discharge through outlet tubes connected to the ports of
reactor vessels received in an apparatus for the synthesis of
multiple organic compounds. The valve means includes first and
second aligned, relatively moveable surfaces between which the
outlet tubes extend. Resilient means are interposed between one of
the surfaces and the outlet tubes. Relative movement of the
surfaces causes force to be applied through the resilient means to
close the outlet tubes.
The surfaces form portions of plates, and more particu-
larly ribs on plates. The resilient means preferrably comprise
Teflon encapsulated silicone O-ring cord sections. One of the rib
surfaces is shaped to correspond to the arcuate shape of the outer
surface of the cord.
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In accordance with. another aspect of the present inven-
tion, apparatus useful for the synthesis of multiple organic com-
pounds is provided. The apparatus includes means adapted to
receive an array of X number of individual reactor vessels, in at
least Y number of different positions, where Y is a number greater
than X. Each of the vessels has a port connected to an outlet
tube. Collection means are provided with an array of 2X number of
collection vessels. Means are provided for shifting the position
of the collection means relative to the vessel receiving means. Hy
first selecting one and then the other set of X number of reactor
vessels of the possible Y number of positions for use, and shifting
the position of the collection means between uses, each of the 2X
number of collection vessels can receive discharge from the outlet
tubes.
The collection means preferrably comprises a standard 96
well microtiter plate. The number X equals 48. The number Y
equals 52. Each of the vessels is adapted to optionally receive a
pores polyethlyene microcannister with a radio frequency trans-
mitter tag.
Valve means are interposed between the reactor vessels
and the collection means for simultaneously regulating the dis-
charge through the outlet tubes.
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In accordance with another aspect of the present inven-
tion, apparatus useful to the synthesis of multiple organic com-
pounds is provided. The apparatus includes means for receiving an
array of tube-like reactor vessels, a plurality of reactor vessels
and optionally a porous polyethelyene microcannister with a radio
frequency transmitter tack for each vessel. Each of the vessels is
adapted to receive a porus polyethelyene microcannister with a
radio frequency transmitter tag.
The vessel receiving means is capable of receiving X
number of reactor vessels in Y number of wells, where Y is a number
is greater than X. The apparatus also includes collection means
having 2X number of collection well . Each of the reactor vessels
has a port connected to an outlet tube. Valve means are provided
for simultaneously regulating the discharge of fluids through the
outlet tubes into the collection wells.
The valve means includes first and second aligned, rela-
tively moveable surfaces between which the outlet tubes extend.
Resilient means are interposed between one of the surfaces and the
outlet tubes. Relative movement of the surfaces causes force to be
applied through the resilient means to close the outlet tubes.
Preferrably, one of the surfaces has an arcuate shape
corresponding to the shape of the exterior of the resilient means.
This maintains the resilient means in proper position.
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In accordance with another aspect of the present inven-
tion, aparatus useful far the synthesis of multiple organic com-
pounds is provided, including a plurality of functional components
stackable in different configurations to fox~n the apparatus. Mul-
tilevel means cooperate with the components to align them. The
alignment means is adapted to be mounted on one of the components.
It has a ffi rst level adapted to support a second component and a
second level adapted to support a third component. The levels are
defined by different sections of the alignment means.
The components include a valve block. The alignment
means is mounted on the valve block.
The second component may include a temperature control
block. A pressure plate is used in conjunction with the tempera-
ture control block.
The third component may including an alignment plate.
The pressure plate is situated above the alignment plate.
The alignment means also includes a bullet nose shaped
section on the top level. This section facilitates assembly of the
blocks.
The alignment means comprises a standoff. Preferrably,
the alignment means includes four standoffs.
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To these and other objects as may hereinafter appear, the
present invention relates to apparatus for the synthesis of mul-
tiple organic compounds with a pinch valve block, as set forth in
detail in the following specification and recited in the annexed
claims, taken together with the accompanying drawings, wherein like
numerals refer to like parts and in which:
Fig. 1 is an isometric view of a first configuration of
the appartus of the present invention including a temperature con-
trol block and showing the collection block in exploded position;
Fig. 2 is an isometric view of a secand configuration of
the apparatus of the present invention without the temperature
control block and with the collection block in exploded position;
Fig. 3 is an exploded isometric view of the reactor block
and valve block of the apparatus of the present invention;
Fig. 4 is an exploded isometric view of the components of
the valve block;
Fig. 5 is a top cross-sectional view of the valve block,
shown in the open state;
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Fig. 6 is an enlarged side cross-sectional view of the
valve block taken along line 6-6 of Fig. 5 showing the reactor
block with the temperature control block;
Fig. 7 is a top cross-sectional view of the valve block,
shown in the closed state;
Fig. 8 is an enlarged cross-sectional view taken along
line 8-8 of Fig. 7;
Fig. 9 is an enlarged exploded side cross-sectional view
of a portion of the plates and slide of the valve block;
Fig. 10 is a view similar to Fig. 6 showing an enlarged
side cross-sectional view of the valve block showing the reactor
block without the temperature control block;
Fig. 11 is a schematic representation of a first set the
selected reactor vessel wells and the set of collection wells with
which they align;
Fig. l2 is a schematic representation of the second set
of the selected reactor vessel wells and the other set of collec-
tion wells with which they align;
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Fig. 13 is a side cross-sectional view of the collection
plate and vacuum adapter in the first relative position; and
Fig. 14 is a view similar to Fig. 13, showing the collec-
tion plate and vacuum adapter in the second relative position.
The apparatus of the present invention relates to a modu-
lar system for the synthesis of diverse organic compounds in which
components in the form of blocks and/or plates are stacked to form
reactors which can be moved among work stations to perform various
steps in the synthesis. A typical reactor consists of a reactor
block, generally designated A, which is adapted to retain a plura-
lity of tube-like reactor vessels 10. Block A may include an
optional temperature control block, generally designated B, as
shown in Figure 1. Reactor Block A is situated above a valve
block, generally designated C, which controls the discharge from
reactor vessels 10 into the collection vessels situated in the
wells 12 of a microtiter plate which forms a portion of a collec-
tion block, generally designated D.
Reactor block A includes a pressure plate 14 with a sep-
tum 16 situated adjacent its undersurface. Pressure plate 14 has
an array of relatively small openings 17, one for each vessel 10.
Openings 17 permit the needle of a syringe to be inserted into the
aligned reactor vessel, through the septum, to introduce liquids
into the vessel. When the temperature control block B is absent,
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as shown in Fig. 2, an alignment plate 18 is situated below septum
16. Alignment plate 18 has an array of openings 19 each of which
receives a reactor vessa_1 10 so as to retain the vessels in the
correct position relative to the pressure plate. Pressure plate 14
is spaced from valve block C so as to permit a plurality of reactor
vessels 10 to the situated therebetween.
Four multi-level alignment standoffs 20 are provided to
retain the components in proper alignment. Standoffs 20 are mount-
ed on valve block C, at each corner of the apparatus. Each stand-
off 20 has a lower, larger diameter section 22, an intermediate,
mid-sized diameter section 24 and a top, bullet shaped section 26.
Temperature control block B, when used (Fig. 1), rests on lower
section 22.
When block B is not used (Fig. 2) , alignment plate 18
rests on the intermediate sections 24 of the standoffs. Alignment
plate 18 is provided with four holes 28 (Fig. 3) . Holes 28 receive
the top portions 26 of standoffs 20. Thus, plate 18 is spaced from
the top surface of the valve block C by the combined height of
sections 22 and 24.
Pressure plate 14 also has four holes 30, somewhat
smaller than holes 28, which receive bullet shaped sections 26 of
standoffs 20. The pressure plate sits over the septum and hence is
spaced above the top surfaces of sections 24 of the standoffs. It
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rests on the rims of the reactor vessels 10, with the septum 16
situated there between (Fig. 10). Clamp brackets 37, located at
either end of the unit, retain pressure plate 14. The bullet shape
of sections 26 of standoffs 20 facilitate positioning of the
plates.
A pair of side latches 32 are mounted on latch pivot
blocks 34 located on each side of the top surface of valve block C.
Each latch has first anal second slots 36, 38 adapted to engage
screws extending from temperature control block B or alignment
plate 18. As shown in F:ig. 1, when temperature control block B is
present, slots 38 on each latch 32 receive the screws extending
from the sides of the temperature control block to latch tempera-
ture control block B in position, when the latches are pivoted to
the upstanding position.. When no temperature control block B is
present, as in Fig. 2, screws from the alignment plate 18 are
received in slots 36.
If control of the temperature of the reactor vessels is
required, block B is interposed between pressure plate 14 and the
valve block C as shown in Fig. 1. Alignment plate 18 is not used
in this configuration. The temperature control block is of con-
ventional design, with an array of vertical reactor vessel receiv-
ing wells and internal conduits through which water or other fluid
can be pumped to regulate the temperature of the reactor vessels.
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Valve block C is illustrated in exploded form in Figs. 4
and 9. This block consists of a top plate 39, a bottom plate 40
and an end cap 42 which, when assembled, define a rectangular
cavity into which a slide 44 is moveably received.
Top plate 39 is provided with openings 46 for screws to
secure it to bottom plate 40. It also has openings 48 for screws
to secure brackets 37 and openings 50 for screws to secure latch
pivot blocks 34.
Further, plate 39 has an array of small holes 52 adapted
to receive the outlet tubes 54 which are attached by Leur tip
adapters 56 to the bottom outlet ports of the reactor vessels 10.
Outlet tubes of this type are commercially available from Supelco,
Inc., Supelco Park, Helleforte, PA 10823 as part No. 5-7059
disposable f low control valve liners. One hole 52 in plate 39 is
provided for each reactor vessel position in reactor block A.
Bottom plate 40 has corresponding holes 58 for receiving
the tubes 54. Holes 58 are of the same number and in the same
locations as holes 52 in plate 39. As best seen in Fig. 4, plate
40 has a "U" shaped configuration, when viewed from the end. The
upper surface of the middle recessed portion 60 of the plate has
eight spaced upstanding ribs 62. Each rib 62 is situated adjacent
a different row of holes 58 and, as best seen in Fig, 9, actually
extends a small way over the rim of the hole.
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Slide 44 also has an array of holes 64 of the same number
and location as holes 52 and holes 58. However, as best seen in
Fig. 9, the top and bottom of each hole 64 is flared outwardly such
that conic sections are formed adjacent the top and bottom surfaces
of slide 44 so as not to cut or permanently distort tubes 54.
The bottom surface of slide 44 is recessed and provided
with eight downwardly extending ribs 66. Each rib 66 is aligned
with a different rib 62 on bottom plate 40. Between each set of
aligned ribs 66, 62 is situated a row of outlet tubes 54. Movement
of slide 44 relative to bottom plate 40 causes ribs 66 to move
towards ribs 62 such that the outlet tubes are pinched closed,
compare Figs. 5, 7 and 8.
However, as is best seen in Figs. 6 and 8, the surface of
rib 66 does not act directly on the walls of the outlet tubes 54.
Instead, it acts through a resilient member 68. Member 68 is
formed of a sections of Teflon encapsulated silicone O-ring cord
commercially available from Lutz Sales Co., Inc. of 4675 Turnbury
Dr.; Hanover Park, Illionis 60103. Member 68 deforms as ribs 66
and 62 are moved toward each other and pinches tubes 54 as seen in
Figs. 7 and 8 in a manlier which does not crush or permanently
deform the wall of the tube. Thus, the tube reliably returns to
its original shape when t:he slide returns to its original position,
as seen in Figs. 5 and 6.
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As best seen in Fig. 9, which is an enlarged exploded
crass-section of a portion of the valve block, the surface of each
rib 66 adjacent the resilient member is arcuate to accomodate_the
curved surface of the resilient member. This curved rib surface
insures the proper positioning the resilient member relative to the
outlet tubes.
Each end of bottom plate 40 is provided with a plurality
of holes 71 each of which is aligned with the space between a dif-
ferent set of ribs 62, 66. Holes 71 extend to the exterior surface
of the plate and have a diameter slightly larger than that of the
resilient members 68. Holes 71 permit resilient members 68 to be
inserted between the ribs 62, 66 after the valve block has been
assembled. Holes 71 may be capped or stopped after the resilient
members are inserted.
Movement of slide 44 relative to plates 40 and 44 is
achieved through the use of end cap 42. End cap 42 has openings 70
to accommodate screws for securing it to bottom and top plates . It
also has a central opening 72 through which a threaded screw 74
with a knob 76 extends. The inner diameter of opening 72 is larger
then the outer diameter of screw 74 such that screw 74 can rotate
freely within opening 72. Screw 76 engages an internally threaded
opening 78 in slide 44 such that rotation of screw 76 in a clock-
wise direction causes slide 44 to move relative to bottom plate 40
such that ribs 66 move towards ribs 62 to close the rows of outlet
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tubes simultaneously. Rotating screw 76 in the counter-clockwise
direction moves the slide to the extreme open posiiton, as shown in
Figs . 5 and 6 such that discharge of fluids through outlet tubes 54
is unimpeded. Overtightening of the plates and crushing or defor-
mation of the tubes is prevented by this structure while complete
closure of all tubes is insured when the valve is closed.
Situated under valve block C is collection block D. As
seen in Fig. 2, block D consists of a locator plate 77 with a
large, generally rectangular central opening 80. Situated below
locator plate 77 is a vacuum adapter 79. Adapter 79 has a central
recess 81 adapted to receive a collection plate, preferrably in the
form of a 96 well densely packed microtiter plate 82 of standard
dimension and configuration. For reasons explained below, locator
plate 77 functions to shift the position of microtiter plate 82
relative to reactor block A. One simple way to accomplish this is
to provide plate 77 in two forms, such that the vacuum adapter is
shifted in position, in one form of the locator plate relative to
the other. In this way, the position of microtiter plate 82 is
shifted relative to the reactor vessels simply by substituting one
form of locator plate 77 for the other.
Liquid is drawn downward through the valve block into the
microtiter plate using vacuum adapter 79 which is commercially
available from Polyfiltronics, Inc. of 100 Weymouth Street,
Rockland MA 02370 which is sold under part number VAC-003. The
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Polyfiltronics vacuum adapter is modified by adding two upstanding
locator pins 83 which fit: into location holes 84 in locator plate
77. Two different locator plates 77 are used alternately to align
the reactor properly over the microtiter plate for product
collection.
Collection block D is situated immediately below valve
block C. The vertical position of collection block D relative to
valve block C can be altered such that different height collection
vessels can be utilized i.n wells 12 of the microtiter plate simply
by addition of the appropriate size spacers (not shown).
Although microtiter plate 82 is capable of receiving 96
collection vessels in i.ts densely packed well array, reactors
designed to accept microcannisters cannot be positioned densely
enough to align with all 96 wells. Hence, two reactors, of slight-
ly different configuration, are normally necessary if all 96 wells
in the microtiter plate are to be used. However, having different
reactor configurations greatly complicates the situation and
increases the cost.
We overcome this problem by using a single reactor in
which 52 possible reactor vessel positions are provided. Thus, two
different sets, of 48 vessel positions each, can be selected. By
changing the position of the microtiter plate relative to the
reactor block, after selecting the second set of 48 vessel posi-
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CA 02380687 2002-04-30
tions for use, discharge into all of the wells in the microtiter
plate can achieved without using two different reactor configura-
tions. This is illustrated schematically in Figs. 11 and 12.
These figures show pressure plate 14 of reactor block A
and microtiter plate 82 of collection block D. The reactor block
A has 52 possible reactor vessel positions, thirteen columns of
four positions each. In Fig. 11, reactor vessels are shown in all
but the right most column. Microtiter plate 82 is shown as posi-
tinned toward the left of the drawings, such that reactors in the
48 occupied positions will discharge into the 48 collection vessels
located in the wells in tike micrvtiter plate designated with a dot .
Referring now to Figure 12, during the next synthesis
set, all of the positions in the reactor block are selected except
the left most column. ~y utilizing a slightly different locator
plate 77, the microtiter plate is shifted to the right, relative to
the reactor vessels. Vessels located in the positions marked by an
~X" discharge into the unused 48 collection vessels in the micro-
titer plate. In this way, the same apparatus, used a second time
after shifting the relative position of the microtiter plate, can
be used to discharge into all 96 wells of a densely packed
microtiter plate.
Figures 13 and 14 illustrate the structure of the two
forms of locator plate 77. With the locator plate shown in Fig.
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CA 02380687 2002-04-30
13, the recess 8l in vacuum adapter 79 containing microtiter plate
82 is shifted in position as compared to recess 81 in the adapter
shown in Fig. 14. Thus, by selecting the appropriate locator plate
form, the microtiter plate will be aligned with either one or the
other set of 48 reactor vessels.
As illustrated in Figure 8, each reactor vessel 10 is
designed so as to receive a commercially available porus polyethe-
lyene microcannister 88 with a radio frequency transmitter tag.
These microcannisters are sold by Irori Quantum Microchemistry of
of 11025 North Torrey Pines Road, LaJolla, CA 92037 and contain the
solid phase support. They are porus so as to permit solutions to
pass through. They are tagged through the use of a microchip with
a semiconductor memory which can store an identification code and
other information relating to the construction of the compound in
the cannister. The tags can be interrogated to obtain the stored
information.
The technique for encoding and tracking combinatorial
chemical libraries with this type of microcannister is disclosed in
a article entitled "Radiofrequency Encoded Combinatorial Chemistry"
by K.C. Nicolaou, Xiao-~t'i Xiao, Zahra Parandoosh, Andrew Senyei and
Michael P. Nova, published in Angew. Chem. Int. Ed. Engl, 1995,
Vol. 34 No. 20 at pages 2289-2291; and an article entitled "Radio
Frequency Tag Encoded Combinatorial Library Method for the Dis-
covery of Tripeptide-Substituted Cinnamic Acid Inhibitors of the
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Protein Tyrosine Phosp'hatase PTP1B" by Edmund Moran, Sepehr
Sarshar, John G. Cargill,, Manouchehr M. Shahbaz, Anna Lio, Adnan M.
M. Mjalli and Robert Armstrong in J. Am. Chem. Soc. 1995, Vol. 117,
No. 43 10787-10788.
One of the advantages of the present invention is that
reactor vessels large enough to receive commercially available
microcannisters with radio frequency transmitter tags for tracking
of the reactors can be employed and a single reactor can be used to
deposit directly into all of the 96 wells of a standard microtiter
plate. This greatly reduces the complexity and cost of the system
while still permitting automated tracking.
It should also be noted that although the apparatus of
the present invention is primarily designed for solid and solution
phase combinatorial synthesis, the reaction vessels could be filled
with a sorb~ent and the products could be purified by passing them
through the sorbent, a process known as "solid phase extraction".
It is believed that the: present design results in a better solid
phase extraction apparatus than anything currently available.
It will now be appreciated that the apparatus of the pre-
sent invention consists of a modular reactor which is simple in
design, small in size, light in weight and inexpensive to build and
operate. It includes a simplied yet extremely reliable valve means
for simultaneously regulating the discharge from the reactor ves-
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CA 02380687 2002-04-30
eels without crushing or deforming the reactor vessel outlet tubes.
The reactor vessels are suitable for receiving porus polyethelene
microcannisters with radio frequency transmitter tags for use in
automated encoding. By providing 52 reactor vessel positions in
the reactor block and a collector block which can shift a standard
96 microtiter plate between two different relative positions, a
single reactor can be used to discharge into all 96 wells of the
microtiter plate. Multilevel alignment standoffs facilitate mount-
ing and alignment of selected components to obtain the desired con-
figuration. Moreover, the apparatus of the present invention can
be used to perform the entire synthesis or only to perform the
cleavage step in a radio frequency tagged synthesis.
while only a limited number of preferred embodiments of
the present invention have been disclosed for purposes of illustra-
tion, it is obvious that many variations and modifications thereof
are possible. It is intended to cover all of these variations and
modifications, which fall within the scope of the present inven-
tion, as recited by the following claims:
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