The present disclosure relates to atypical split N- and C-inteins and variants thereof. This disclosure also relates to complexes comprising the split N- or C-inteins of this disclosure and a compound of interest and compositions comprising said complexes. In addition, this disclosure relates to methods of using the atypical split N- and C-inteins.

The object of the present invention is to provide a method for manufacturing an amino acid polymer more simply and efficiently compared to conventional methods for manufacturing amino acid polymers.

The present invention provides a method for manufacturing an amino acid polymer with thioacid amino acids. Specifically, the manufacturing method of the present invention comprises (A) a step of preparing first and second thioacid amino acids, (B) a step of subjecting said first and second thioacid amino acids to an oxidation reaction to obtain an amino acid polymer linked by peptide bonds. The manufacturing method of the present invention is characterized in that it partially uses thioacid amino acids that do not possess a protecting group.

The present invention has an object of providing a peptide synthesis method using a carrier capable of reversibly repeating the dissolved state and the insolubilized state, wherein the problem of an amino acid active species existing in the reaction system in de-protection reaction can be easily solved. The present invention provides a peptide synthesis method comprising the following steps: a step of condensing an N-Fmoc protected amino acid with a peptide having a C-terminal protected with a carrier which is crystallized according to a change of a composition of a dissolving solvent, in the presence of a condensing agent, to obtain an N-Fmoc-C-carrier protected peptide, a step of adding an alkylamine having 1 to 14 carbon atoms or hydroxyl amine to the reaction system, a step of de-protecting the N-terminal, and a step of changing the composition of the solvent dissolving the C-carrier protected peptide, to crystallize and separate the peptide.

The present invention provides a method for preparing a PNA oligomer. More specifically, the present invention can prepare a PNA oligomer which is easily separable from byproducts through a simple and short process by using PNA dimers, PNA trimers or PNA tetramers, and which has extremely high yields and purity.

A method for synthesizing a peptide having the sequence His-X-Glu-Gly-Thr-Phe-Thr-Ser-Asp-Val-Ser-Ser-Tyr-Leu-Glu-Gly-Gln-Ala-Ala-Y-Glu-Phe-Ile-Ala-Trp-Leu-Val-Z-Gly-Arg-Gly is disclosed. The method includes enzymatically coupling: (a) a peptide C-terminal ester or thioester having a first peptide fragment with the sequence His-X-Glu-Gly-Thr-Phe-Thr-Ser-Asp-Val-Ser-(thio)ester; and (b) a peptide nucleophile having an N-terminally unprotected amine having a second peptide fragment with the sequence H-Ser-Tyr-Leu-Glu-Gly-Gln-Ala-Ala-Y-Glu-Phe-Ile-Ala-Trp-Leu-Val-Z-Gly-Arg-Gly; wherein: X is Ala or an -amino-isobutyric acid (Aib) residue; Y is Lys, which Lys has a free side-chain -amino group or of which Lys the side-chain -amino group is protected with a protective group or of which Lys the side-chain -amino group is functionalized with an amino acid or another functional group; and Z is Arg or Lys.

A method for synthesizing a peptide having the sequence His-X-Glu-Gly-Thr-Phe-Thr-Ser-Asp-Val-Ser-Ser-Tyr-Leu-Glu-Gly-Gln-Ala-Ala-Y-Glu-Phe-Ile-Ala-Trp-Leu-Val-Z-Gly-Arg-Gly is disclosed. The method includes enzymatically coupling: (a) a peptide C-terminal ester or thioester having a first peptide fragment with the sequence His-X-Glu-Gly-Thr-Phe-Thr-Ser-Asp-Val-Ser-(thio)ester; and (b) a peptide nucleophile having an N-terminally unprotected amine having a second peptide fragment with the sequence H-Ser-Tyr-Leu-Glu-Gly-Gln-Ala-Ala-Y-Glu-Phe-Ile-Ala-Trp-Leu-Val-Z-Gly-Arg-Gly; wherein: X is Ala or an -amino-isobutyric acid (Aib) residue; Y is Lys, which Lys has a free side-chain -amino group or of which Lys the side-chain -amino group is protected with a protective group or of which Lys the side-chain -amino group is functionalized with an amino acid or another functional group; and Z is Arg or Lys.

In certain embodiments a device is provided for electrorotation flow. In certain embodiments the device comprises a microfluidic channel comprising a plurality of electrodes disposed to provide dielectrophoretic (DEP) forces that are perpendicular to hydrodynamic flows along the channel; and a fluid within the channel providing the hydrodynamic flow along the channel; wherein the device is configured to apply focusing voltages to the electrodes that provide an electric field minimum in the channel and that focus cells, particles, and/or molecules or molecular complexes within the channel; and where the device is configured to apply rotation-inducing voltages to the electrodes that induce rotation of the cells, particles, molecules and/or molecular complexes as they flow through the channel.

The instant disclosure is directed to solution phase fragment coupling methods for preparing etelcalcetide and its pharmaceutically acceptable salts.

A method for synthesizing a peptide having the sequence His-X-Glu-Gly-Thr-Phe-Thr-Ser-Asp-Val-Ser-Ser-Tyr-Leu-Glu-Gly-Gln-Ala-Ala-Y-Glu-Phe-Ile-Ala-Trp-Leu-Val-Z-Gly-Arg-Gly is disclosed. The method includes enzymatically coupling: (a) a peptide C-terminal ester or thioester having a first peptide fragment with the sequence His-X-Glu-Gly-Thr-Phe-Thr-Ser-Asp-Val-Ser-(thio)ester; and (b) a peptide nucleophile having an N-terminally unprotected amine having a second peptide fragment with the sequence H-Ser-Tyr-Leu-Glu-Gly-Gln-Ala-Ala-Y-Glu-Phe-Ile-Ala-Trp-Leu-Val-Z-Gly-Arg-Gly; wherein: X is Ala or an -amino-isobutyric acid (Aib) residue; Y is Lys, which Lys has a free side-chain -amino group or of which Lys the side-chain -amino group is protected with a protective group or of which Lys the side-chain -amino group is functionalized with an amino acid or another functional group; and Z is Arg or Lys.

It was found that a salt formed of an acid and a base having characteristics set forth below can inactivate a deprotecting agent, thereby suppressing redundant peptide elongation: (i) the base is different in type from a base used as a deprotecting agent, and (ii) a conjugate acid of the base has a pKa smaller than that of a conjugate acid of a base used as a deprotecting agent.