# US 2020/0399306 A1 · High shear solid phase synthesis

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- Number: US 2020/0399306 A1
- Document ID: US20200399306A1
- Status: Published application
- Kind: A1
- Application number: 16/979,212
- Filed: 2019-03-12
- Published: 2020-12-24
- Earliest claimed: 2018-03-13
- Assignees: Yissum Research Development Company of the Hebrew University of Jerusalem
- Applicants: Yissum Research Development Company of the Hebrew University of Jerusalem Ltd.
- Inventors: Chaim GILON; Moshe BENTOLILA
- Web page: https://patentgrep.com/patent/US20200399306A1

## CPC

- C07K 1/045: {using devices to improve synthesis, e.g. reactors, special vessels}
- B01J 19/00: Chemical, physical or physico-chemical processes in general; Their relevant apparatus
- 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)}
- C07K: PEPTIDES (peptides in foodstuffs A23; obtaining protein compositions for foodstuffs, working-up proteins for foodstuffs A23J; preparations for medicinal purposes A61K; peptides containing beta-lactam rings C07D; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, C07D; ergot alkaloids of the cyclic peptide type C07D519/02; macromolecular compounds having statistically distributed amino acid units in their molecules, i.e. when the preparation does not provide for a specific; but for a random sequence of the amino acid units, homopolyamides and block copolyamides derived from amino acids C08G69/00; macromolecular products derived from proteins C08H1/00; preparation of glue or gelatine C09H; single cell proteins, enzymes C12N; genetic engineering processes for obtaining peptides C12N15/00; compositions for measuring or testing processes involving enzymes C12Q; investigation or analysis of biological material G01N33/00)
- B01J: CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS

## Abstract

The present disclosure relates to solid phase synthesis of organic molecules and particularly to highly efficient methods for synthesizing polymers, such as peptides, nucleotides or saccharides, employing solid phase synthesis.

## Claims

### Claim at position 1

1-51. (canceled)

### Claim 52

A method for performing at least one cycle of solid phase synthesis of an organic molecule, the method comprising the steps of:
i. providing a reactor comprising a reaction chamber and a stirring apparatus, wherein the stirring apparatus comprises an impeller having at least two blades rotatable about an axis;
ii. inserting beads of functionalized polymeric resin and at least one solvent into the reactor to provide a reaction mixture, wherein the reaction mixture is in contact with the rotatable blades;
iii. inserting at least one reactant into the reaction chamber;
iv. spinning the impeller for a period of time, at a rotational rate of at least 600 rounds per minute (rpm), while maintaining a shear rate of at least 3·103 sec−1;
thereby performing at least one cycle of the solid phase synthesis of the organic molecule.

### Claim 53 (depends on 52)

The method of claim 52, wherein the stirring apparatus is a mechanical stirrer and spinning of the impeller is performed at a rotational rate of 600 to 1400 rounds per minute, maintaining a shear rate of at least 3·103 sec−1.

### Claim 54 (depends on 53)

The method of claim 53, wherein the spinning of the impeller comprises maintaining shear stress of at least 1.5 N/m2 in the reaction mixture.

### Claim 55 (depends on 52)

The method of claim 52, wherein the stirring apparatus is a homogenizer and spinning of the impeller is performed at a rotational rate of 5,000-30,000 rounds per minutes, maintaining a shear rate of at least 1·106 sec−1.

### Claim 56 (depends on 55)

The method of claim 55, wherein the homogenizer is a rotor-stator homogenizer.

### Claim 57 (depends on 52)

The method of claim 52, wherein the functionalized beads of polymeric resin have particle size of 20-200 μm.

### Claim 58 (depends on 52)

The method of claim 52, wherein the organic molecule is a polymer comprising a molecule selected from the group consisting of a peptide chain, a nucleotide chain and a sugar.

### Claim 59 (depends on 52)

The method of claim 52, wherein the organic molecule comprises a peptide chain.

### Claim 60 (depends on 58)

The method of claim 58, for performing a cycle in the solid phase synthesis of a polymeric organic molecule, wherein the method comprises the steps of:
(a) providing a reactor comprising a reaction chamber and a stirring apparatus comprising an impeller having at least two blades rotatable about an axis;
(b) inserting beads of functionalized polymeric resin and at least one solvent into the reactor to provide a reaction mixture, wherein the reaction mixture is in contact with the rotatable blades;
(c) inserting at least one protected monomeric organic molecule and at least one coupling agent into the reaction chamber and spinning the impeller, thereby forming a coupling product of the protected monomeric organic molecule and the resin;
(d) washing excess of the protected monomeric organic molecule; and
(e) inserting at least one deprotecting reagent into the reaction chamber and spinning the impeller, thereby removing at least one protecting group from the coupling product, forming a coupling product of the deprotected monomeric organic molecule and the resin, thereby completing a cycle in the solid phase synthesis of a polymeric organic molecule;
wherein the spinning of the impeller in at least one of steps (c) and (e) is performed for a period of time, at a rotational rate of at least 600 rounds per minute, while maintaining a shear rate of at least 3·103 sec−1, optionally wherein steps (c) to (e) are repeated a plurality of cycles.

### Claim 61 (depends on 60)

The method of claim 60, wherein both steps (c) and (e) are performed for a period of time, at a rotational rate of at least 600 rounds per minute, while maintaining a shear rate of at least 3·103 sec−1.

### Claim 62 (depends on 58)

The method of claim 58, wherein the at least one reactant is selected from the group consisting of: a deprotection agent, a coupling agent, and a protected monomeric organic molecule.

### Claim 63 (depends on 58)

The method of claim 58, wherein the at least one reactant comprises a coupling agent and a protected monomeric organic molecule, thereby performing at least one coupling cycle of the solid phase synthesis of the organic molecule.

### Claim 64 (depends on 63)

The method of claim 63, wherein the protected monomeric organic molecule is an α-N-protected amino acid.

### Claim 65 (depends on 58)

The method of claim 58, wherein the reactant comprises at least one deprotecting reagent.

### Claim 66 (depends on 52)

The method of claim 52, further comprising the steps of washing the reaction mixture and filtering the beads of polymeric resin.

### Claim 67 (depends on 52)

The method of claim 52, further comprising repeating steps (ii) to (iv) at least one more time thereby performing at least one additional cycle of the solid phase synthesis of the organic molecule.

### Claim 68 (depends on 67)

The method of claim 67, comprising at least two steps of coupling of an amino acid to the resin and at least two steps of removal of protecting group.

### Claim 69 (depends on 58)

The method of claim 58, further comprising the step of cleavage of the polymeric organic molecule from the polymeric resin.

### Claim 70 (depends on 52)

The method of claim 52, comprising an initial step of swelling the beads of polymeric resin in at least one solvent.

### Claim 71 (depends on 70)

The method of claim 70, wherein swelling the beads of polymeric resin comprises mixing the beads of polymeric resin for a specified period of time in the solvent, at a rotational rate of at least 600 rounds per minute and maintaining shear rate of at least 3·103 sec−1.