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EP-4736894-A1 - ANTIMICROBIAL PEPTIDE-FUNCTIONALIZED CROSSLINKABLE COPOLYMER, CROSSLINKED COPOLYMER AND HYDROGEL WITH ANTIBACTERIAL ACTIVITY

EP4736894A1EP 4736894 A1EP4736894 A1EP 4736894A1EP-4736894-A1

Abstract

The present invention relates to a crosslinkable copolymer, the crosslinkable copolymer comprising a hydrophilic monomer, a monomer functionalized with a photocrosslinkable group and a monomer functionalized with an antimicrobial peptide that is covalently bound via at least one of its amino groups. The present invention further relates to a crosslinked copolymer and a hydrogel derived therefrom, as well as a wound dressing comprising one or more of the crosslinkable polymer(s), the crosslinked copolymer or the hydrogel.

Inventors

  • JONAS, ULRICH
  • ATIF, MUHAMMAD
  • ZAAT, Sebastian A.J.
  • Gizem, Babuccu

Assignees

  • Universität Siegen, Körperschaft des öffentlichen Rechts
  • University Medical Center Amsterdam

Dates

Publication Date
20260506
Application Date
20241031

Claims (14)

  1. A crosslinkable copolymer, the crosslinkable copolymer comprising a hydrophilic monomer, a monomer functionalized with a photocrosslinkable group and a monomer functionalized with an antimicrobial peptide that is covalently bound via at least one of its amino groups.
  2. The crosslinkable copolymer according to claim 1, wherein the crosslinkable copolymer comprises a monomer functionalized with a chromogenic dye convertible by a bacterial enzyme, wherein the chromogenic dye optionally is selected from the group comprising 4-nitrophenyl β-D-glucuronide, 5-bromo-4-chloro-3-indolyl-β-D-glucuronide, 4-methylumbelliferyl β-D-glucuronide and 4-methylumbelliferyl α-D-glucuronide, and wherein the monomer optionally is N-(6-aminohexyl)acrylamide.
  3. The crosslinkable copolymer according to claim 1 or 2, wherein the crosslinkable copolymer comprises a monomer functionalized with a surfactant, wherein the surfactant optionally is selected from the group comprising ethylene oxide-propylene oxide copolymer mono(2-ethylhexyl) ethers, poly(oxyethylene) lauryl ethers and 4-(1,1,3,3-tetramethylbutyl)phenyl-polyethylene glycols, and wherein the monomer optionally is selected from the group comprising acrylate and methacrylate.
  4. The crosslinkable copolymer according to any one of the preceding claims, wherein the antimicrobial peptide is selected from the group of peptides comprising acetyl-LKRVWKRVFKLLKRYWRQLKKPVR (SAAP-148) (SEQ ID NO: 1), acetyl-LKRLYKRLAKLIKRLYRYLKKPVR-amide (SAAP-145) (SEQ ID NO: 2), acetyl-LKRVWKAVFKLLKRYWRQLKKPVR-amide (SAAP-276) (SEQ ID NO: 3), acetyl-IGKEFKRIVERIKRFLRELVRPLR-amide (OP-145) (SEQ ID NO: 4), LRCMCIKWWSGKHPK-amide (TC19) (SEQ ID NO: 5), LRAMCIKWWSGKHPK-amide (TC84) (SEQ ID NO: 6) and GKWKLFKKAFKKFLKILAC (BP2) (SEQ ID NO: 7).
  5. The crosslinkable copolymer according to any one of the preceding claims, wherein the hydrophilic monomer is selected from the group of monomers comprising hydroxyethyl acrylamide, hydroxypropyl acrylamide, N-vinyl-2-pyrrolidone, N,N-dimethylacrylamide, methacrylamide, 2-hydroxyethyl methacrylate, 2-hydroxypropyl methacrylate, 2-methyl-2-oxazoline, 2-ethyl-2-oxazoline, 2-n-propyl-2-oxazoline and 2-isopropyl-2-oxazoline.
  6. The crosslinkable copolymer according to any one of the preceding claims, wherein the monomer functionalized with a photocrosslinkable group is selected from (4-benzoylphenyl)acrylamide and 4-benzoylphenyl)methacrylamide, and optionally is N-(4-benzoylphenyl)acrylamide.
  7. The crosslinkable copolymer according to any one of the preceding claims, wherein the antimicrobial peptide binds to an acrylate monomer functionalized with pentafluorophenole via reaction of at least one of its amino group(s) forming an amide bond with the acrylate.
  8. The crosslinkable copolymer according to any one of the preceding claims, wherein the crosslinkable polymer has the following formula (1): wherein: A is an antimicrobial peptide selected from the group of SAAP-148 (SEQ ID NO: 1), SAAP-145 (SEQ ID NO: 2), SAAP-276 (SEQ ID NO: 3), OP-145 (SEQ ID NO: 4), TC19 (SEQ ID NO: 5), TC84 (SEQ ID NO: 6) and BP2 (SEQ ID NO: 7), B is a chromogenic dye selected from the group comprising 4-nitrophenyl β-D-glucuronide (PNPG), 5-bromo-4-chloro-3-indolyl-β-D-glucuronide (X-Gluc), 4-methylumbelliferyl β-D-glucuronide (MUG) and 4-methylumbelliferyl α-D-glucuronide (MUD), E is a surfactant selected from the group of ethylene oxide-propylene oxide copolymer mono(2-ethylhexyl) ethers, poly(oxyethylene) lauryl ethers and 4-(1,1,3,3-tetramethylbutyl)phenyl-polyethylene glycols, x is in a range of 0.6 to 0.95, preferably 0.88, y is in a range of 0.01 to 0.03, preferably 0.02, z is in a range of 0 to 0.10, preferably 0.01 to 0.10, more preferably 0.07, m is in a range of 0.0001 to 0.1, preferably 0.001, n is in a range of 0 to 0.03, preferably 0.01 to 0.03, more preferably 0.02.
  9. The crosslinkable copolymer according to any one of the preceding claims, wherein the crosslinkable polymer is selected from the group of the following polymers: wherein x is in a range of 0.6 to 0.95, preferably 0.88, y is in a range of 0.01 to 0.03, preferably 0.02, z is in a range of 0.01 to 0.10, preferably 0.07, m is in a range of 0.0001 to 0.1, preferably 0.001, and n is in a range of 0.01 to 0.03, preferably 0.02, wherein x is in a range of 0.6 to 0.95, preferably 0.88, y is in a range of 0.01 to 0.03, preferably 0.02, z is in a range of 0.01 to 0.10, preferably 0.07, m is in a range of 0.0001 to 0.1, preferably 0.001, n is in a range of 0.01 to 0.03, preferably 0.02. and
  10. A crosslinked copolymer, obtained by photo-induced crosslinking of one or more of the crosslinkable polymer(s) according to any one of claims 1 to 9.
  11. A hydrogel, comprising the crosslinked copolymer according to claim 10.
  12. A wound dressing, comprising one or more of the crosslinkable polymer(s) according to any one of claims 1 to 9, the crosslinked copolymer according to claim 10 or the hydrogel according to claim 11.
  13. The hydrogel according to claim 11 or the wound dressing according to claim 12 for use in the diagnosis and/or treatment of a bacterial infection.
  14. The hydrogel or the wound dressing for use according to claim 13, wherein the bacterial infection is caused by Escherichia coli (E. coli) , Staphylococcus aureus , Pseudomonas aeruginosa (P. aeruginosa) and / or Acinetobacter baumanii (A. baumanii).

Description

The present invention relates to a crosslinkable copolymer and a crosslinked copolymer that can form a hydrogel, which is usable as wound dressing in the diagnosis and/or treatment of bacterial infections. Healthcare-Associated Infections (HAIs) arise within medical facilities, where a most severe healthcare-associated infection is wound infection. The presence of bacterial infections in wounds, which are often difficult to detect in an early stage, can significantly impede the healing process. Among the various microorganisms responsible for healthcare-associated infections, Escherichia coli (E. coli) has been identified as a cause of most severe infections. Various wound dressings that are usable in the diagnosis or treatment of bacterial infections are known, such as wound dressings based on hydrogels. CN 11135341 A describes an intelligent hydrogel dressing for early warning and controllable treatment of infected wound surfaces. The hydrogel consists of four parts: polyvinyl alcohol which is biocompatible and is often used as wound surface dressing, nanoparticles which can perceive pH changes of wound surfaces and have fluorescence resonance energy transferring effects, macromolecules responding to ultraviolet rays and capable of controllably releasing antibiotics, and upper transformation nanoparticles. CN 114159615 A describes a hydrogel wound dressing that can indicate the pH of the wound and is also responsive to ultrasound for drug delivery. The dressing is made up of a pH-sensitive hydrogel matrix that changes color when there is a change in the pH of the wound. In addition, the hydrogel matrix is loaded with drug nanoparticles that are responsive to ultrasound. When ultrasound is applied to the wound, the drug nanoparticles are released from the hydrogel matrix, resulting in enhanced drug delivery to the wound site. CA 2644432 A1 describes a wound dressing that contains both an anti-inflammatory pain-killing agent and a complex of silver ion and a transitional element of Group IV of the periodic system of elements. The wound dressing can provide effective pain relief and anti-inflammatory benefits while also offering antimicrobial properties due to the presence of silver ions. The transitional element of Group IV of the periodic system of elements is added to enhance the antimicrobial properties of the silver ions. US 2009209897 A1 describes a photoactivated antimicrobial wound dressing comprising a photocatalytic membrane. The membrane comprises a bacterial cellulose hydrogel membrane with a photosensitizer coated on its surface. When the photosensitizer is activated by light, it produces reactive oxygen species that can kill bacteria on a wound. Wound dressings for bacterial monitoring comprising sensor elements or electronic devices have been described. US 2021/0000409 A1 describes a sensor system for non-implantable medical devices that can detect and identify microbiological infections. The system uses a combination of sensing technologies to provide real-time detection of pathogens and can be integrated with wireless communication for data transmission. WO 2018/211458 A1 describes a wound covering that allows for wound monitoring and therapeutic agent delivery. The wound covering comprises a sensor element that undergoes a change in appearance in response to a change in a wound, and a therapeutic agent delivery system for delivering therapeutic agents to the wound site. The sensor element comprises fibers comprising a hydrogel and sensor particles. US 10702153 B2 describes a wound dressing that includes reusable electronics for wireless monitoring of the wound. The dressing can monitor various aspects of the wound, such as temperature, moisture, and pH levels. The electronics are attached to the dressing using a snap-fit mechanism, making them easy to attach and remove. Also, chromogenic substrates for the identification of bacteria are known. WO 2018/060711 A1 discloses a device for detecting the presence of microorganisms in wounds. A genetically engineered microbial sensor, such as Resazurim, 10-acetyl-3,7-dihydroxyphenoxazine or a tetrazolium salt, produces a detectable signal in response to the presence of a target bacterium, which signal can be measured using various detection techniques, such as fluorescence, luminescence, or electrochemical sensing. CN 109439725 A describes a chromogenic medium containing one chromogenic substrate of 4-nitrophenyl-beta-D-glucopyranoside, o-nitrophenol beta-D glucuronide, 4-nitrophenyl-beta-D-glucopyranoside, 4-methylumbelliferyl-beta-D-glucuronide and 5-bromo-4-chloro-3-indoxyl-N-acetyl-beta-D-glucosaminide. The chromogenic substrate interacts with glucosaccharase of Escherichia coli to release a chromogenic group. Applications in food testing are described. 5-Bromo-4-chloro-1H-indol-3-yl-β-D-glucopyranosiduronic acid (X-Gluc) is commercially available for the colorimetric detection of glucuronidase activity, such as via Carl Roth GmbH & Co. KG. Chi