Patents Assigned to Enzypep B.V.
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Patent number: 10920258Abstract: 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.Type: GrantFiled: February 6, 2020Date of Patent: February 16, 2021Assignee: ENZYPEP B.V.Inventors: Timo Nuijens, Ana Toplak, Peter Jan Leonard Mario Quaedflieg
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Patent number: 10883132Abstract: 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.Type: GrantFiled: March 11, 2019Date of Patent: January 5, 2021Assignee: ENZYPEP B.V.Inventors: Timo Nuijens, Ana Toplak, Peter Jan Leonard Mario Quaedflieg
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Patent number: 10858414Abstract: The invention relates to a method for preparing a coupling product having the sequence Pq-Wv-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.Type: GrantFiled: March 8, 2019Date of Patent: December 8, 2020Assignee: ENZYPEP B.V.Inventors: Timo Nuijens, Ana Toplak, Peter Jan Leonard Mario Quaedflieg
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Publication number: 20200347427Abstract: 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.Type: ApplicationFiled: February 6, 2020Publication date: November 5, 2020Applicant: ENZYPEP B.V.Inventors: Timo NUIJENS, Ana TOPLAK, Peter Jan Leonard Mario QUAEDFLIEG
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Patent number: 10752931Abstract: The invention further relates to a process for the enzymatic synthesis of an (oligo)peptide. The invention relates to a method for designing an enzymatic synthesis process of an (oligo)peptide, comprising identifying two or more (oligo)peptide fragments of an (oligo)peptide, which fragments are (oligo)peptides suitable for preparing the (oligo)peptide by enzymatic condensation of the two or more peptide fragments using a ligase. The invention relates to a method for designing an enzymatic synthesis process of a cyclic (oligo)peptide, comprising identifying a non-cyclic (oligo)peptide from which the cyclic (oligo)peptide can be prepared by cyclisation, catalysed by a cyclase. The invention further relates to a process for the enzymatic synthesis of an (oligo)peptide.Type: GrantFiled: July 8, 2016Date of Patent: August 25, 2020Assignee: ENZYPEP B.V.Inventors: Timo Nuijens, Peter Jan Leonard Mario Quaedflieg
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Publication number: 20200262886Abstract: The invention relates to a method for preparing a coupling product having the sequence Pq-Wv-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.Type: ApplicationFiled: March 8, 2019Publication date: August 20, 2020Applicant: ENZYPEP B.V.Inventors: Timo NUIJENS, Ana TOPLAK, Peter Jan Leonard Mario QUAEDFLIEG
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Publication number: 20190345529Abstract: 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.Type: ApplicationFiled: March 11, 2019Publication date: November 14, 2019Applicant: ENZYPEP B.V.Inventors: Timo NUIJENS, Ana TOPLAK, Peter Jan Leonard Mario QUAEDFLIEG
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Patent number: 10336994Abstract: The invention relates to a subtilisin BPN? variant or homolog thereof, having the following mutations compared to subtilisin BPN? represented by SEQUENCE ID NO: 2 or a homolog sequence thereof:—a deletion of the amino acids corresponding to positions 75-83;—a mutation at the amino acid position corresponding to S221, the mutation corresponding to S221C or S221 selenocysteine, preferably S221C;—at least one further mutation selected from the group consisting of amino acid positions corresponding to F189W, F189Y, S33D, S33T, N218D, N218T, N218E, N62D, N62S, N62W, and N62Y; preferably a mutation at the amino acid position corresponding to P225; wherein the amino acid positions are defined according to the sequence of subtilisin BPN? represented by SEQUENCE ID NO: 2. The invention further relates to enzymatically synthesizing a peptide.Type: GrantFiled: November 16, 2018Date of Patent: July 2, 2019Assignee: ENZYPEP B.V.Inventors: Ana Toplak, Timo Nuijens, Peter Jan Leonard Mario Quaedflieg
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Patent number: 10253061Abstract: The invention relates to a method for enzymatically synthesizing an (oligo)peptide, comprising coupling (a) an (oligo)peptide C-terminal ester or thioester and (b) an (oligo)peptide nucleophile having an N-terminally unprotected amine, wherein the coupling is carried out in a fluid comprising water, and wherein the coupling is catalyzed by a subtilisin BPN? variant or a homolog thereof, which comprises the following mutations compared to subtilisin BPN? represented by SEQUENCE ID NO: 2 or a homolog sequence thereof: a deletion of the amino acids corresponding to positions 75-83; a mutation at the amino acid position corresponding to S221, the mutation being S221C or S221 selenocysteine; preferably a mutation at the amino acid position corresponding to P225 wherein the amino acid positions are defined according to the sequence of subtilisin BPN? represented by SEQUENCE ID NO: 2. Further, the invention relates to an enzyme suitable for use as a catalyst in a method of the invention.Type: GrantFiled: October 17, 2018Date of Patent: April 9, 2019Assignee: ENZYPEP B.V.Inventors: Peter Jan Leonard Mario Quaedflieg, Timo Nuijens, Jan Metske Van Der Laan, Dirk Barend Janssen, Ana Toplak, Bian Wu
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Patent number: 10138268Abstract: The invention relates to a method for enzymatically synthesizing an (oligo)peptide, comprising coupling (a) an (oligo)peptide C-terminal ester or thioester and (b) an (oligo)peptide nucleophile having an N-terminally unprotected amine, wherein the coupling is carried out in a fluid comprising water, and wherein the coupling is catalyzed by a subtilisin BPN? variant or a homo-log thereof, which comprises the following mutations compared to subtilisin BPN? represented by SEQUENCE ID NO: 2 or a homolog sequence thereof: a deletion of the amino acids corresponding to positions 75-83; a mutation at the amino acid position corresponding to S221, the mutation being S221C or S221 selenocysteine; preferably a mutation at the amino acid position corresponding to P225 wherein the amino acid positions are defined according to the sequence of subtilisin BPN? represented by SEQUENCE ID NO: 2. Further, the invention relates to an enzyme suitable for use as a catalyst in a method of the invention.Type: GrantFiled: October 9, 2015Date of Patent: November 27, 2018Assignee: ENZYPEP B.V.Inventors: Peter Jan Leonard Mario Quaedflieg, Timo Nuijens, Jan Metske Van Der Laan, Dirk Barend Janssen, Ana Toplak, Bian Wu
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Publication number: 20170305963Abstract: The invention relates to a method for enzymatically synthesizing an (oligo)peptide, comprising coupling (a) an (oligo)peptide C-terminal ester or thioester and (b) an (oligo)peptide nucleophile having an N-terminally unprotected amine, wherein the coupling is carried out in a fluid comprising water, and wherein the coupling is catalyzed by a subtilisin BPN? variant or a homologue thereof, which comprises the following mutations compared to subtilisin BPN? represented by SEQUENCE ID NO: 2 or a homologue sequence thereof: a deletion of the amino acids corresponding to positions 75-83; a mutation at the amino acid position corresponding to S221, the mutation being S221C or S221 selenocysteine; preferably a mutation at the amino acid position corresponding to P225 wherein the amino acid positions are defined according to the sequence of subtilisin BPN? represented by SEQUENCE ID NO: 2. Further, the invention relates to an enzyme suitable for use as a catalyst in a method of the invention.Type: ApplicationFiled: October 9, 2015Publication date: October 26, 2017Applicant: ENZYPEP B.V.Inventors: Peter Jan Leonard Mario QUAEDFLIEG, Timo NUIJENS, Jan Metske VAN DER LAAN, Dirk Barend JANSSEN, Ana TOPLAK, Bian WU
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Patent number: 9598714Abstract: Method for enzymatically synthesizing an oligopeptide, comprising the coupling of an (optionally N-protected) protected oligopeptide ester with an (optionally C-protected) protected oligopeptide nucleophile in an organic solvent or an organic solvent mixture having a water content of 0.1 vol % or less, by a subtilisin in any possible form.Type: GrantFiled: February 28, 2013Date of Patent: March 21, 2017Assignee: ENZYPEP B.V.Inventors: Peter Jan Leonard Mario Quaedflieg, Timo Nuijens
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Publication number: 20150037840Abstract: Method for enzymatically synthesising an oligopeptide, comprising the coupling of an (optionally N-protected) protected oligopeptide ester with an (optionally C-protected) protected oligopeptide nucleophile in an organic solvent or an organic solvent mixture having a water content of 0.1 vol % or less, by a subtilisin in any possible form.Type: ApplicationFiled: February 28, 2013Publication date: February 5, 2015Applicant: EnzyPep B.V.Inventors: Peter Jan Leonard Mario Quaedflieg, Timo Nuijens
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Patent number: 8883444Abstract: The invention relates to a method for synthesizing a peptide by enzymatically preparing an ester or thioester from (i) an N-terminal protected amino acid or an N-terminal protected peptide where either can have a protected C-terminal ester group and (ii) an alcohol represented by the formula HO—CX2—Z or a thiol represented by the formula HS—CX2—Z, each X independently representing a halogen atom or a hydrogen atom; and Z represents an electron withdrawing group comprising at least one sp3-hybridized carbon comprising at least two substituents comprising a heteroatom directly attached to the at least one sp3-hybridized carbon or at least one sp2-hybridized carbon comprising one or two substituents comprising a heteroatom directly attached to the at least one sp2-hybridized carbon, and enzymatically coupling the prepared ester or thioester with an optionally C-terminal protected amino acid or with an optionally C-terminal protected peptide in a medium comprising 2 wt. % water or less.Type: GrantFiled: November 19, 2009Date of Patent: November 11, 2014Assignee: Enzypep B.V.Inventors: Peter Jan Leonard Mario Quaedflieg, Timo Nuijens, Claudia Cusan, Catharina Hubertina Maria Schepers
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Patent number: 8450084Abstract: The invention relates to a method for preparing an optionally N-protected amino acid C-terminal ester or an optionally N-protected peptide C-terminal ester, comprising transesterifying the C-terminal t-alkyl ester of the amino acid or the C-terminal t-alkyl ester of the peptide with an alcohol (other than the t-alcohol corresponding to the t-alkyl group of the ester) in the presence of a hydrolytic enzyme (E.C. 3). The invention further relates to a method for preparing a peptide comprising coupling an activated, N-protected, amino acid C-terminal ester or an optionally N-protected peptide C-terminal ester with an optionally C-terminal protected amino acid or an optionally C-terminal protected peptide via a peptide bond, in the presence of an enzyme catalysing peptidic bond formation.Type: GrantFiled: October 9, 2008Date of Patent: May 28, 2013Assignee: Enzypep B.V.Inventors: Peter Jan Leonard Mario Quaedflieg, Timo Nuijens, Claudia Cusan, Harold Monro Moody, Theodorus Johannes Godfried Maria Van Dooren