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US-20260125736-A1 - FURIN SENSORS AND APPLICATIONS THEREOF

US20260125736A1US 20260125736 A1US20260125736 A1US 20260125736A1US-20260125736-A1

Abstract

Diagnostic furin sensors, respective furin sensitive peptides and related kits are disclosed, in addition to methods and medical uses involving the sensors, peptides or kits, such as methods for diagnosing cancer (particularly HNSCC), in a subject, as well as to methods for determining furin activity.

Inventors

  • Lorenz Meinel
  • Marcus Otto GUTMANN
  • Debora REINHARDT
  • Björn TER MORS
  • Marc DRIESSEN

Assignees

  • FLAREON BIOTECH GMBH

Dates

Publication Date
20260507
Application Date
20251107
Priority Date
20230508

Claims (20)

  1. 1 . A diagnostic sensor comprising: a quenching moiety R1, a Furin-sensitive peptide Pep, and a signaling moiety R2, wherein the quenching moiety R1 suppresses the signal from the signaling moiety R2 in the said sensor.
  2. 2 . The sensor of claim 1 , A) wherein the Furin-sensitive peptide Pep comprises amino acid residues having the sequence P4-P3-P2-P1-P1′-P2′-P3′-P4′ (SEQ ID NO: 17), wherein P4 is independently selected from G and R, P3 is T, P2 is A, P1 is independently selected from M and G, P1′ is independently selected from T, G and S, P2′ is A, P3′ is G, and P4′ is A; or B) wherein the Furin-sensitive peptide Pep comprises amino acid residues having the sequence RRARSVAS (SEQ ID NO: 1) or SSARSVAS (SEQ ID NO: 2).
  3. 3 . The sensor of claim 1 , further comprising, a first additional moiety L1, a second additional moiety L2.
  4. 4 . The sensor of claim 3 , wherein the sensor has a structure selected from the group consisting of R1-L1-Pep-L2-R2, L1-R1-Pep-L2-R2, L1-R1-Pep-R2-L2, and R1-L2-Pep-R2-L2.
  5. 5 . The sensor of claim 1 , wherein the quenching moiety R1 comprises a moiety selected from the group consisting of commercially available quenching moieties.
  6. 6 . The sensor of claim 5 , wherein R1 comprises 4-((4-(Dimethylamino)phenyl)azo)benzoic acid.
  7. 7 . The sensor of claim 5 , wherein R1 is acetylated.
  8. 8 . The sensor of claim 5 , wherein R1 comprises Ac-K(Dabcyl).
  9. 9 . The sensor of claim 1 , wherein the fluorophore R2 comprises a moiety selected from the group consisting of small organic dyes, fluorescent proteins, and quantum dots.
  10. 10 . The sensor of claim 1 , wherein R2 comprises 5-((2-Aminoethyl)amino)naphthalene-1-sulfonic acid.
  11. 11 . The sensor of claim 1 , wherein R2 comprises -E(Edans).
  12. 12 . The sensor of claim 3 , wherein A) L1 comprises a moiety selected from the group consisting of linkers, natural polymers and synthetic polymers, or B) L2 comprises a moiety selected from the group consisting of linkers, natural polymers and synthetic polymers.
  13. 13 . A furin-sensitive peptide having a length of from 8 to 16 amino acids, wherein A) the Furin-sensitive peptide Pep comprises amino acid residues having the sequence P4-P3-P2-P1-P1′-P2′-P3′-P4′ (SEQ ID NO: 17), wherein P4 is independently selected from G and R, P3 is T, P2 is A, P1 is independently selected from M and G, P1′ is independently selected from T, G and S, P2′ is A, P3′ is G, and P4′ is A; or B) the Furin-sensitive peptide Pep comprises amino acid residues having the sequence RRARSVAS (SEQ ID NO: 1) or SSARSVAS (SEQ ID NO: 2).
  14. 14 . A diagnostic kit comprising the sensor in accordance with claim 1 .
  15. 15 . A diagnostic kit comprising the peptide in accordance with claim 13 .
  16. 16 . A method of diagnosing cancer in a subject, wherein the method comprises using a sensor in accordance with claim 1 , wherein a sample has been obtained from the subject, wherein an increased level of furin activity relative to the level of a control is indicative of the subject having cancer; wherein the method comprises a) determining a level of furin activity in the sample, b) comparing the level in the said sample to a control level of furin activity, c) diagnosing whether the subject has cancer, wherein an increased level of furin activity is indicative of the subject having cancer.
  17. 17 . The method of claim 16 , wherein the sample has been obtained from the oral cavity of the subject.
  18. 18 . The method of claim 16 , wherein the subject is a human subject.
  19. 19 . The method of claim 16 , wherein the cancer is a head and neck cancer.
  20. 20 . The method of claim 19 , wherein the cancer is HNSCC.

Description

CROSS-REFERENCES TO RELATED APPLICATIONS This application is a continuation of copending international patent application PCT/EP2024/062630 filed on 7 May 2024 and designating the United States of America, which was published under Article 21(2) PCT in English, and which claims priority of European patent application EP 23172136.6 filed on 8 May 2023, all of which are herewith incorporated herein by reference. FIELD OF THE INVENTION The present invention relates to the field of cancer, in particular head and neck cancer, such as squamous cell carcinomas of the head and neck (“HNSCC”, “head and neck squamous cell carcinoma”). REFERENCE TO SEQUENCE LISTING A Sequence Listing submitted as an XML file via Patent Center is hereby incorporated by reference in accordance with 35 U.S.C. § 1.52(e). The name of the XML file for the Sequence Listing is 62378522_1.XML, the date of creation of the XML file is Nov. 7, 2025, and the size of the XML file is 22,022 bytes. BACKGROUND Head and neck cancer is a major global health problem. It shows a 5-year survival rate of about two-thirds, while ranking among the top 10 most prevailing cancers worldwide [1, 2]. The malignancies are mostly defined as squamous cell carcinomas of the head and neck (HNSCC), which typically arise in the oral cavity, nasal cavity, pharynx and larynx [3]. Main risk factors for development of HNSCC include smoking, alcohol consumption and human papillomavirus (HPV) infection [4]. The diagnosis of HNSCC is complex and often delayed, as most patients with HNSCC (over 60%) are diagnosed in later stages (Ill or IV), resulting in poorer survival rates for the effected patients, bringing up the need for earlier detection of HNSCC [5, 6]. Common methods for diagnosis include physical examination, radiographic findings, and biopsies of the tissues of interest, resulting in a TNM-classification, separated into different categories from I to IV. The stage is based on the anatomic extent of the tumor, lymph node involvement, and metastasis (“TNM”: “Tumor, lymph node, metastasis”) [5, 7]. TNM-classification of the tumor dictates treatment options, involving multiple specialists, that handle a wide variety of therapy options including surgery, radiotherapy, chemotherapy, chemoradiotherapy, small molecule therapy and targeted therapies e.g., anti-EGFR therapy with Cetuximab [8, 9]. Among others, innovations in surgery have improved HNSCC therapy significantly over the last dec-ades and continuing these trends is desirable [1, 10]. The key to reaping the benefits of the various treatments is considered the (i) early detection and (ii) broad applicability/accessibility. Hence, there is a general need in the art to improve diagnosis accordingly. Specifically, as squamous cell carcinomas of the head and neck (HNSCC) are one of most prevailing cancers worldwide and present a massive global health burden, Therapy of the malignancies would e.g. benefit greatly if overall diagnosis was shifted to an earlier stage. Consequently, there generally is a need in the art to provide for earlier diagnosis. Moreover, particularly also in the context of head and neck cancer, there is a need in the art for further improvements of diagnosis, such as by addressing a broader population or making testing easier for medical professionals or even at the individual level. In general, to this end point-of-care-testing (POCT) does not only offer the possibility to approach diagnostic problems via non-invasive testing, but can also address a broader population, compared to traditional diagnostic procedures performed by medical professionals [20]. The uncomplicated application paired with the simple readout makes POCT an effective tool in different medical situations ranging from infectious diseases to cancer diagnosis [21, 22]. POCT systems can be used on the individual level or by medical professionals as bed-side testing, to acquire faster diagnostic information compared to traditional laboratory-based methods. Furin is a ubiquitous transmembrane endoprotease. It is involved in a broad range of physiological cellular functions, as well as involvement in multiple diseases, ranging from COVID-19 to various cancer types [11, 12]. Furins cleavage motif in physiological conditions is described as Arg-X-Lys/Arg-Arg 1 [13, 14]. Many pathogens, however, use this consensus sequence to exploit the extracellular protease function for maturation of their envelope proteins to enable infection of the host cell [15]. In cancer on the other hand, furins aberrant activity has been described as an important point to malignancies via proteolytic activation of e.g., hormones, receptors and growth factors [16] and elevated expression or increased activity of furin has been studied to some extent in connection with cancer [17-19]. FRET techniques (where FRET stands for “F6rster resonance energy transfer”, which is often also referred to “fluorescence resonance energy transfer” even though it is not restricted to