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KR-20260066182-A - MATRIX METALLOPROTEASE-CLEAVABLE AND SERINE OR CYSTEINE PROTEASECLEAVABLE SUBSTRATES AND METHODS OF USE THEREOF

KR20260066182AKR 20260066182 AKR20260066182 AKR 20260066182AKR-20260066182-A

Abstract

The present invention relates to a polypeptide comprising, in general, at least a first cleavable moiety (CM1) which is a substrate for at least one matrix metalloproteinase (MMP) and at least a second cleavable moiety (CM2) which is a substrate for at least one serine protease (SP) or at least one cysteine protease (CP); an activable antibody; other larger molecules comprising such polypeptide comprising at least CM1 which is a substrate for at least one MMP protease and at least CM2 which is a substrate for at least one SP protease or at least one cysteine protease (CP); and methods for preparing and using such polypeptide comprising at least CM1 which is a substrate for at least one MMP protease and at least CM2 which is a substrate for at least one SP protease or at least one cysteine protease (CP) in various therapeutic, diagnostic, and prophylactic indications.

Inventors

  • 바실리예바 올가
  • 윈터 마이클 비.

Assignees

  • 싸이톰스 테라퓨틱스, 인크.

Dates

Publication Date
20260512
Application Date
20191205
Priority Date
20181206

Claims (14)

  1. A cleavable polypeptide comprising a cleavable moiety (CM) comprising an amino acid sequence selected from the group consisting of GLPTFVHL (SEQ No. 135) and GLPTFVH (SEQ No. 136).
  2. In claim 1, the cleavable polypeptide wherein the CM comprises the amino acid sequence of GLPTFVHL (SEQ ID NO. 135).
  3. In claim 1, the cleavable polypeptide wherein the CM comprises the amino acid sequence of GLPTFVH (SEQ No. 136).
  4. A cleavable polypeptide according to claim 1, comprising an amino acid sequence selected from the group consisting of ALAHGLPTFVHLGGGS (SEQ No. 235), GLPTFVHLPRQVGGGS (SEQ No. 236), ALAHGLPTFVHLGGS (SEQ No. 250), and GLPTFVHLPRQVGGS (SEQ No. 251).
  5. A cleavable polypeptide comprising the amino acid sequence of SEQ ID NO. 459 or 460 in claim 1.
  6. A composition comprising the cleavable polypeptide and carrier of claim 1.
  7. A conjugated polypeptide comprising the cleavable polypeptide of claim 1 conjugated to an agent.
  8. In claim 7, the above formulation is a conjugated polypeptide selected from the group consisting of the following: (a) Toxin or fragment of it; (b) microtubule inhibitor; (c) nucleic acid damage agents; (d) dolastatin or a derivative thereof; (e) Auristatin or a derivative thereof; (f) maytansinoid or a derivative thereof; (g) Duocarmycin or a derivative thereof; (h) calicheamicin or a derivative thereof; (i) pyrrolobenzodiazepine or a derivative thereof; (j) Auristatin E or a derivative thereof; (k) monomethylauristatin E (MMAE); (l) Monomethylauristatin D (MMAD); (m) Meitansinoid DM1; (n) Meitansinoid DM4; (o) Detectable area; and (p) Diagnostic preparations.
  9. A cleavable polypeptide comprising a cleavable site (CM) comprising an amino acid sequence selected from the group consisting of SEQ ID NOs 226-255.
  10. In claim 9, the cleavable polypeptide comprising an amino acid sequence selected from the group consisting of SEQ ID NOs 226-240.
  11. In claim 9, the cleavable polypeptide comprising an amino acid sequence selected from the group consisting of SEQ ID NOs 241-255.
  12. In claim 9, the cleavable polypeptide comprising an amino acid sequence selected from the group consisting of SEQ ID NOs 226-234.
  13. In claim 9, the cleavable polypeptide comprising an amino acid sequence selected from the group consisting of SEQ ID NOs 237-249.
  14. In claim 9, the cleavable polypeptide comprising an amino acid sequence selected from the group consisting of SEQ ID NOs 252-255.

Description

Matrix Metalloprotease-Cleavable Substrates and Serine or Cysteine Protease-Cleavable Substrates and Methods of Use Thereof Cross-reference regarding related applications This application claims priority to U.S. provisional application No. 62/776,409 filed December 6, 2018 and No. 62/778,062 filed December 11, 2018, the contents of which are incorporated herein by reference in their entirety. Technology field The present invention generally involves at least one matrix metalloprotease: At least a first cleavable moiety (CM1) that is a substrate for MMPs and at least one serine protease The invention relates to a polypeptide comprising at least a second cleavable moiety (CM2) that is a substrate for SP and/or at least one cysteine protease (CP), an activable antibody comprising at least CM1 that is a substrate for at least one MMP protease and CM2 that is a substrate for at least one SP protease and/or at least one CP protease, and other larger molecules and a method for preparing and using such polypeptide comprising at least CM1 that is a substrate for at least one MMP protease and CM2 that is a substrate for at least one SP protease and/or at least one CP protease in various therapeutic, diagnostic and prophylactic indications. Reference to the sequence list The “Sequence List,” which was electronically submitted concurrently with this application in a computer-readable format (CFR) via EFS-Web under the filename “CYTX-058-PCT_ST25” in accordance with 37 C.F.R. § 1.821, is incorporated herein by reference. An electronic copy of the Sequence List was created on November 26, 2019, and has a disk size of 159 kilobytes. Proteases are enzymes that break down proteins by cleaving peptide bonds between amino acid residues. Proteases occur naturally in all organisms and are involved in a wide range of physiological responses, from simple degradation to highly regulated pathways. Some proteases are known to break specific peptide bonds based on the presence of specific amino acid sequences within a protein. Therefore, it is necessary to identify new substrates for protease and utilize these substrates for various therapeutic, diagnostic, and prophylactic indications. In an embodiment of the present invention, an isolated polypeptide comprising a tandem substrate is provided herein, wherein the tandem substrate comprises at least a first cleavable moiety (CM1) which is a substrate for at least one matrix metalloproteinase (MMP) and at least a second cleavable moiety (CM2) which is a substrate for at least one serine protase (SP) or cysteine protase (CP), and CM1 comprises the amino acid sequence AHGL or PRQV, and the N-terminal to C-terminal arrangement of the tandem substrate is CM1-CM2 or CM2-CM1. In some embodiments, CM1 of the isolated polypeptide comprises an amino acid sequence selected from the group consisting of ALAHGLF (SEQ No. 1), ALAHGL (SEQ No. 52), LAHGLF (SEQ No. 50), LAHGL (SEQ No. 53), and AHGLF (SEQ No. 51). In some embodiments, CM1 of the isolated polypeptide comprises an amino acid sequence selected from the group consisting of HVPRQV (SEQ No. 8) and VPRQV (SEQ No. 60). In some embodiments, the isolated polypeptide of the present disclosure comprises CM1 and CM2 connected by a linking peptide. In some embodiments, CM1 and CM2 of the isolated polypeptide are directly connected to each other. In some embodiments, the isolated polypeptide of the present disclosure comprises CM2 comprising a substrate for a CP enzyme, wherein the CP enzyme is legumain. In some embodiments, the isolated polypeptide of the present disclosure comprises CM2 comprising a substrate for an SP enzyme selected from the group consisting of urokinase, matriptase, and neutrophil elastase. In some embodiments, the isolated polypeptide of the present disclosure comprises CM2 comprising a substrate for an SP enzyme selected from the group consisting of urokinase, matrixtase, and neutrophil elastase, and a substrate for a CP enzyme, wherein the CP enzyme is legumine. In some embodiments, the isolated polypeptide of the present disclosure comprises CM1 comprising a substrate for an MMP enzyme selected from the group consisting of MMP2, MMP9, or MMP14. In some embodiments, the isolated polypeptide of the present disclosure comprises CM2 having an amino acid sequence selected from the group consisting of SGR, LSGR (SEQ No. 73), ARG, PRS, TFVH (SEQ No. 141), AAN, SAN, and GPTN (SEQ No. 152). In some embodiments, the isolated polypeptide of the present disclosure comprises an amino acid sequence selected from the group consisting of SGR, LSGR (SEQ No. 73), LSGRS (SEQ No. 72), LSGRSD (SEQ No. 71), LSGRSA (SEQ No. 110), LSGRSDN (SEQ No. 70), LSGRSAN (SEQ No. 109), LSGRSDNH (SEQ No. 20), LSGRSGNH (SEQ No. 78), LSGRSDNP (SEQ No. 90), LSGRSDNI (SEQ No. 84), LSGRSANI (SEQ No. 108), LSGRSANP (SEQ No. 114), LSGRSDYH (SEQ No. 86), LSGRSDTH (SEQ No. 92), LSGRSDQH (SEQ No. 96), LSGRSDIH (SEQ No. 100), and LSGRSDDH (SEQ No. 104). Includes CM2. In s