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CN-122029182-A - Methods, compositions, and kits comprising cell penetrating agents

CN122029182ACN 122029182 ACN122029182 ACN 122029182ACN-122029182-A

Abstract

The present disclosure provides cell penetrating agents comprising a cell internalization module and a payload, and methods of delivering a payload to a cell using the cell penetrating agents.

Inventors

  • T.S. Nijar
  • S. J. Tam

Assignees

  • 普罗塞纳平台技术有限公司

Dates

Publication Date
20260512
Application Date
20240913
Priority Date
20230915

Claims (20)

  1. 1. A Cell Membrane Internalization Peptide (CMIP) comprising an amino acid sequence selected from the group consisting of formula (IA) to formula (IH): X 1 -X 2 -X 3 -X 4 -X 5 -X 6 -X 7 -F-S-G-K-a-a-A-K-X 16 -E-A-K-X 20 -X 21 -X 22 -X 23 -X 24 -X 25 (IA); I-W-L-T-A-L-X 7 -F-X 9 -X 10 -X 11 -X 12 -X 13 -X 14 -X 15 -A-X 17 -X 18 -X 19 -Q-F-X 22 -X 23 -X 24 -X 25 (IB); I-W-X 3 -X 4 -X 5 -X 6 -K-X 8 -S-X 10 -X 11 -H-X 13 -X 14 -X 15 -A-E-X 18 -X 19 -X 20 -X 21 -L-S-K-L (IC); X 1 -W-X 3 -T-A-X 6 -X 7 -X 8 -X 9 -G-K-A-X 13 -A-K-A-X 17 -A-K-Q-X 21 -X 22 -S-K-X 25 (ID); X 2 -X 3 -X 4 -X 5 -L-K-F-X 9 -X 10 -X 11 -X 12 -a-a-K-X 16 -E-X 18 -K-Q-F-X 22 -X 23 -X 24 -L (IE); X 1 -W-L-T-A-X 6 -X 7 -X 8 -S-G-X 11 -X 12 -a-a-X 15 -A-X 17 -X 18 -K-X 20 -F-X 22 -X 23 -X 24 -X 25 (IF); S-X 2 -L-T-A-X 6 -X 7 -X 8 -X 9 -X 10 -K-H-X 13 -I-T-Q-X 17 -X 18 -K-R-R-X 22 -X 23 -X 24 -X 25 (IG); R-R-L-T-X 5 -L-F-K-X 9 -G-X 11 -X 12 -X 13 -X 14 -E-X 16 -I-X 18 -X 19 -X 20 -F (IH); Wherein the method comprises the steps of X 1 、X 2 、X 3 、X 4 、X 5 、X 6 、X 7 、X 8 、X 9 、X 10 、X 11 、X 12 、X 13 、X 14 、X 15 、X 16 、X 17 、X 18 、X 19 、X 20 And X 21 are each independently a natural or unnatural amino acid residue, and X 22 、X 23 、X 24 and X 25 are each independently absent or are each independently a natural or unnatural amino acid residue; Or a pharmaceutically acceptable salt thereof.
  2. 2. The CMIP or salt of claim 1, wherein at least one of (a) to (c) is present in the CMIP sequence: (a) X 21 is selected from alanine, arginine, lysine, phenylalanine, and tryptophan; (b) X 9 is serine and X 12 is selected from alanine, arginine, glutamic acid, glutamine, phenylalanine and tryptophan, or (C) X 5 is histidine and X 3 is selected from alanine, glutamic acid, leucine, lysine, phenylalanine and tryptophan.
  3. 3. A CMIP or salt as claimed in claim 1 or claim 2, wherein the CMIP consists essentially of an amino acid sequence selected from formulae (IA) to (IH).
  4. 4. A CMIP or salt as claimed in any one of claims 1 to 3, wherein the CMIP consists of an amino acid sequence selected from formulae (IA) to (IH).
  5. 5. The CMIP or salt of any one of claims 1-4, wherein the CMIP comprises formula (IA): X 1 -X 2 -X 3 -X 4 -X 5 -X 6 -X 7 -F-S-G-K-a-a-A-K-X 16 -E-A-K-X 20 -X 21 -X 22 -X 23 -X 24 -X 25 (IA) Or a pharmaceutically acceptable salt thereof.
  6. 6. The CMIP or salt of any one of claims 1-4, wherein the CMIP comprises formula (IB): I-W-L-T-A-L-X 7 -F-X 9 -X 10 -X 11 -X 12 -X 13 -X 14 -X 15 -A-X 17 -X 18 -X 19 -Q-F-X 22 -X 23 -X 24 -X 25 (IB); Or a pharmaceutically acceptable salt thereof.
  7. 7. The CMIP or salt of any one of claims 1-4, wherein the CMIP comprises formula (IC): I-W-X 3 -X 4 -X 5 -X 6 -K-X 8 -S-X 10 -X 11 -H-X 13 -X 14 -X 15 -A-E-X 18 -X 19 -X 20 -X 21 -L-S-K-L (IC); Or a pharmaceutically acceptable salt thereof.
  8. 8. The CMIP or salt of any one of claims 1-4, wherein the CMIP comprises formula (ID): X 1 -W-X 3 -T-A-X 6 -X 7 -X 8 -X 9 -G-K-A-X 13 -A-K-A-X 17 -A-K-Q-X 21 -X 22 -S-K-X 25 (ID); Or a pharmaceutically acceptable salt thereof.
  9. 9. The CMIP or salt of any one of claims 1-4, wherein the CMIP comprises formula (IE): X 2 -X 3 -X 4 -X 5 -L-K-F-X 9 -X 10 -X 11 -X 12 -a-a-K-X 16 -E-X 18 -K-Q-F-X 22 -X 23 -X 24 -L (IE); Or a pharmaceutically acceptable salt thereof.
  10. 10. The CMIP or salt of any one of claims 1-4, wherein the CMIP comprises formula (IF): X 1 -W-L-T-A-X 6 -X 7 -X 8 -S-G-X 11 -X 12 -a-a-X 15 -A-X 17 -X 18 -K-X 20 -F-X 22 -X 23 -X 24 -X 25 (IF); Or a pharmaceutically acceptable salt thereof.
  11. 11. The CMIP or salt of any one of claims 1-4, wherein the CMIP comprises formula (IG): S-X 2 -L-T-A-X 6 -X 7 -X 8 -X 9 -X 10 -K-H-X 13 -I-T-Q-X 17 -X 18 -K-R-R-X 22 -X 23 -X 24 -X 25 (IG); Or a pharmaceutically acceptable salt thereof.
  12. 12. The CMIP or salt of any one of claims 1-4, wherein the CMIP comprises formula (IH): R-R-L-T-X 5 -L-F-K-X 9 -G-X 11 -X 12 -X 13 -X 14 -E-X 16 -I-X 18 -X 19 -X 20 -F (IH); Or a pharmaceutically acceptable salt thereof.
  13. 13. A Cell Membrane Internalization Peptide (CMIP) comprising an amino acid sequence selected from formulas (IIA) to (IID): X 2 -X 3 -T-A-L-X 7 -F-X 9 -G-X 11 -a-a-X 14 -K-X 16 -E-A-X 19 -Q-F-L-X 23 -X 24 -L (IIA); I-W-X 3 -X 4 -A-L-X 7 -F-X 9 -G-X 11 -X 12 -X 13 -A-X 15 -A-E-A-X 19 -X 20 -F-X 22 -S-X 24 -L (IIB); X 1 -X 2 -L-T-X 5 -L-K-X 8 -S-X 10 -K-a-a-A-X 15 -A-E-A-K-Q-X 21 -L-S-X 24 -L (IIC); I-W-L-T-X 5 -X 6 -K-F-S-X 10 -K-X 12 -a-a-K-A-X 17 -X 18 -K-Q-F-L-X 23 -X 24 -X 25 (IID); Wherein the method comprises the steps of X 1 、X 2 、X 3 、X 4 、X 5 、X 6 、X 7 、X 8 、X 9 、X 10 、X 11 、X 12 、X 13 、X 14 、X 15 、X 16 、X 17 、X 18 、X 19 、X 20 、X 21 And X 22 are each independently a natural or unnatural amino acid residue; And X 23 、X 24 and X 25 are either absent or each independently a natural or unnatural amino acid residue; Or a pharmaceutically acceptable salt thereof.
  14. 14. The CMIP or salt of claim 13, wherein the CMIP consists essentially of an amino acid sequence selected from formulas (IIA) to (IID).
  15. 15. The CMIP or salt of claim 13 or claim 14, wherein the CMIP consists of an amino acid sequence selected from formulae (IIA) to (IID).
  16. 16. The CMIP or salt of any one of claims 13-15, wherein the CMIP comprises formula (IIA): X 2 -X 3 -T-A-L-X 7 -F-X 9 -G-X 11 -a-a-X 14 -K-X 16 -E-A-X 19 -Q-F-L-X 23 -X 24 -L (IIA); Or a pharmaceutically acceptable salt thereof.
  17. 17. The CMIP or salt of any one of claims 13-15, wherein the CMIP comprises formula (IIB): I-W-X 3 -X 4 -A-L-X 7 -F-X 9 -G-X 11 -X 12 -X 13 -A-X 15 -A-E-A-X 19 -X 20 -F-X 22 -S-X 24 -L (IIB); Or a pharmaceutically acceptable salt thereof.
  18. 18. The CMIP or salt of any one of claims 13-15, wherein the CMIP comprises formula (IIC): X 1 -X 2 -L-T-X 5 -L-K-X 8 -S-X 10 -K-a-a-A-X 15 -A-E-A-K-Q-X 21 -L-S-X 24 -L (IIC); Or a pharmaceutically acceptable salt thereof.
  19. 19. The CMIP or salt of any one of claims 13-15, wherein the CMIP comprises formula (IID): I-W-L-T-X 5 -X 6 -K-F-S-X 10 -K-X 12 -a-a-K-A-X 17 -X 18 -K-Q-F-L-X 23 -X 24 -X 25 (IID); Or a pharmaceutically acceptable salt thereof.
  20. 20. The CMIP or salt of any one of claims 13-19, wherein the polypeptide comprises at least three positively charged amino acid residues, each occupying a position corresponding to X 7 、X 11 、X 15 、X 19 or X 24 .

Description

Methods, compositions, and kits comprising cell penetrating agents Cross Reference to Related Applications The present application claims priority from U.S. provisional patent application No. 63/538,596 filed on 9/15 of 2023, the contents of which are hereby incorporated by reference in their entirety. Sequence listing The present application contains a sequence table that has been submitted electronically as an XML file named "50887-0007WO1_ST26_SL.XML". The XML file was created at 2024, 8, 30, and size 238,166 bytes. The content in the XML file is hereby incorporated by reference in its entirety. Background Biomolecules and macromolecules are valuable assets for diagnostic and therapeutic applications. Antibodies alone have become a major therapeutic modality with remarkable success in the treatment of a variety of diseases, including cancer, immune disorders and infectious diseases. Over the last forty years, more than one hundred antibody-based drugs have been FDA approved for the treatment of these diseases. However, many biological molecules and macromolecules (including antibodies) have limited ability to enter cells. Thus, therapeutic and diagnostic applications of these molecules are largely limited by binding to and/or modulating extracellular targets, such as cell surface receptors. Although success has been achieved to date by modulating extracellular targets, intracellular targets also play a critical role in disease progression and progression. Treatments based on modulating these intracellular targets have traditionally utilized small molecules. However, many intracellular targets are extremely difficult to modulate with small molecules, and in some cases, these targets are referred to in the literature as "non-patentable". By facilitating the intracellular delivery of payloads that bind and/or modulate these intracellular targets, the therapeutic and diagnostic potential of biomolecules and/or macromolecules can be greatly expanded for many biomedical applications, including diagnostics, therapeutics, and basic research. Current intracellular delivery systems have not had significant success in delivering payloads directed against intracellular targets. Thus, new systems and methods are needed to facilitate intracellular delivery of payload molecules. Disclosure of Invention Provided herein is a Cell Membrane Internalization Peptide (CMIP) comprising an amino acid sequence selected from the group consisting of formulas (IA) to (IH): X1-X2-X3-X4-X5-X6-X7-F-S-G-K-a-a-A-K-X16-E-A-K-X20-X21-X22-X23-X24-X25(IA); I-W-L-T-A-L-X7-F-X9-X10-X11-X12-X13-X14-X15-A-X17-X18-X19-Q-F-X22-X23-X24-X25(IB); I-W-X3-X4-X5-X6-K-X8-S-X10-X11-H-X13-X14-X15-A-E-X18-X19-X20-X21-L-S-K-L (IC); X1-W-X3-T-A-X6-X7-X8-X9-G-K-A-X13-A-K-A-X17-A-K-Q-X21-X22-S-K-X25(ID); X2-X3-X4-X5-L-K-F-X9-X10-X11-X12-a-a-K-X16-E-X18-K-Q-F-X22-X23-X24-L (IE); X1-W-L-T-A-X6-X7-X8-S-G-X11-X12-a-a-X15-A-X17-X18-K-X20-F-X22-X23-X24-X25(IF); S-X2-L-T-A-X6-X7-X8-X9-X10-K-H-X13-I-T-Q-X17-X18-K-R-R-X22-X23-X24-X25(IG); R-R-L-T-X5-L-F-K-X9-G-X11-X12-X13-X14-E-X16-I-X18-X19-X20-F (IH); Wherein X1、X2、X3、X4、X5、X6、X7、X8、X9、X10、X11、X12、X13、X14、X15、X16、X17、X18、X19、X20 and X 21 are each independently a natural or unnatural amino acid residue, and X 22、X23、X24 and X 25 are each independently absent or are each independently a natural or unnatural amino acid residue, or a pharmaceutically acceptable salt thereof. In some embodiments, at least one of (a) to (c) is present in the CMIP sequence: (a) X 21 is selected from alanine, arginine, lysine, phenylalanine, and tryptophan; (b) X 9 is serine and X 12 is selected from alanine, arginine, glutamic acid, glutamine, phenylalanine and tryptophan, or (C) X 5 is histidine and X 3 is selected from alanine, glutamic acid, leucine, lysine, phenylalanine and tryptophan. In some aspects, the present disclosure provides a Cell Membrane Internalization Peptide (CMIP) comprising an amino acid sequence selected from the group consisting of formulas (IA) to (IH): X1-X2-X3-X4-X5-X6-X7-F-S-G-K-a-a-A-K-X16-E-A-K-X20-X21-X22-X23-X24-X25(IA); I-W-L-T-A-L-X7-F-X9-X10-X11-X12-X13-X14-X15-A-X17-X18-X19-Q-F-X22-X23-X24-X25(IB); I-W-X3-X4-X5-X6-K-X8-S-X10-X11-H-X13-X14-X15-A-E-X18-X19-X20-X21-L-S-K-L (IC); X1-W-X3-T-A-X6-X7-X8-X9-G-K-A-X13-A-K-A-X17-A-K-Q-X21-X22-S-K-X25(ID); X2-X3-X4-X5-L-K-F-X9-X10-X11-X12-a-a-K-X16-E-X18-K-Q-F-X22-X23-X24-L (IE); X1-W-L-T-A-X6-X7-X8-S-G-X11-X12-a-a-X15-A-X17-X18-K-X20-F-X22-X23-X24-X25(IF); S-X2-L-T-A-X6-X7-X8-X9-X10-K-H-X13-I-T-Q-X17-X18-K-R-R-X22-X23-X24-X25(IG); R-R-L-T-X5-L-F-K-X9-G-X11-X12-X13-X14-E-X16-I-X18-X19-X20-F (IH); Wherein each of X 1 to X 25 is independently an amino acid residue selected from the group consisting of alanine, arginine, asparagine, aspartic acid, cysteine, glutamic acid, glutamine, glycine, histidine, isoleucine, leucine, lysine, methionine, phenylalanine, proline, serine, threonine, tryptophan, tyrosine and valine, or a pharmaceutically acceptable salt thereof. In so