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EP-4118431-B1 - INHIBITION OF FKBP1A FOR THE TREATMENT OF TRIPLE-NEGATIVE MAMMARY CARCINOMA

EP4118431B1EP 4118431 B1EP4118431 B1EP 4118431B1EP-4118431-B1

Inventors

  • ESSER, Knud
  • KULIK, Andrea
  • FEHM, TANJA
  • STEIMEL, Judith

Dates

Publication Date
20260506
Application Date
20210311

Claims (15)

  1. A method for finding inhibitors of peptidyl-prolyl cis-trans isomerase FKBP1A that are suitable for treating triple-negative breast cancer, comprising the steps of (a) providing an inhibitor (i) of the expression of FKBP1A; and/or (ii) of the enzymatic activity of FKBP1A; and/or (iii) of the interaction(s) of FKBP1A with interaction partner(s); (b) incubating cancer cells of a cancer cell line with the inhibitor of FKBP1A, and determining a property of the cancer cells selected from the group consisting of (A) cellular differentiation status, (B) epithelial properties, (C) cellular immunogenicity, (D) apoptosis induction or apoptosis capacity, and (E) cellular stem cell character; (c) incubating cancer cells of the same cancer cell line as in step (b) in the absence of the FKBP1A inhibitor and determining the same property of the cancer cells as in step (b) under the same conditions as step (b); (d) comparing the determined property of the cancer cells according to steps (b) and (c).
  2. The method according to claim 1, wherein step (a) comprises the sub-steps (a 1 ) providing a plurality of test substances; (a 2 ) screening the test substances for their inhibitory effect on (i) the expression of FKBP1A; and/or (ii) the enzymatic activity of FKBP1A; and/or (iii) interaction(s) of FKBP1A with interaction partner(s); (a 3 ) selecting at least one screened test substance whose inhibitory effect is stronger than the inhibitory effect of at least one other screened test substance, and providing this selected test substance as an inhibitor of FKBP1A.
  3. The method according to any of the preceding claims, wherein the inhibitor provided in step (a) - inhibits the interaction(s) of FKBP1A with interaction partner(s) and wherein at least one interaction partner is selected from the group consisting of interaction partners which reduce or arrest (A) the cellular differentiation status, (B) the epithelial properties, (C) the cellular immunogenicity, (D) the apoptosis induction or apoptosis capacity, and/or (E) the cellular stem cell character; - inhibits the interaction(s) of FKBP1A with interaction partner(s), wherein at least one interaction partner is selected from the group consisting of interacting proteins, interacting peptides, and interacting nucleic acids; - inhibits the expression of FKBP1A and not simultaneously (ii) the enzymatic activity of FKB1A; and/or - (ii) inhibits the enzymatic activity of FKB1A and simultaneously does not inhibit (i) the expression of FKBP1A.
  4. The method according to any of the preceding claims, wherein - in step (b), the property is determined by quantifying differentiation markers, epithelial markers, (tumor) stem cell markers, immunomodulatory proteins, immune-mediating proteins, or combinations thereof; - step (b) comprises incubating a mixture of cancer cells of a cancer cell line and immune cells of an immune cell line with the inhibitor of FKBP1A, and wherein the property of the cancer cells is determined as their cellular immunogenicity by quantifying immune cell activation and/or by quantifying the immune cell-mediated cytotoxic effect on the cancer cells; and wherein step (c) comprises incubating a mixture of cancer cells of the same cancer cell line and immune cells of the same immune cell line as in step (b) in the absence of the FKBP1A inhibitor and determining the same property of the cancer cells as in step (b) under the same conditions as step (b); - the property determined in step (b) is (A) cellular differentiation status; preferably wherein in step (b) the property is determined by quantifying mesenchymal markers; preferably by quantifying vimentin; - the property (C) determined in step (b) is cellular immunogenicity; preferably wherein in step (b) the property is determined by quantifying immunomodulatory proteins; preferably by quantifying PD-L1; - the property (E) determined in step (b) is cellular stem cell character; preferably wherein in step (b) the property is determined by quantifying (tumor) stem cell markers; preferably by quantifying CD44/CD24 and ALDH1; and/or - the property determined in step (b) is the quantification of the expression of FKBP1A.
  5. The method according to any of the preceding claims, comprising the steps (a) providing an inhibitor (i) of the expression of FKBP1A; preferably wherein step (a) comprises the sub-steps (a 1 ) providing a plurality of test substances; (a 2 ) screening the test substances for their inhibitory effect on (i) the expression of FKBP1A; (a 3 ) selecting at least one screened test substance whose inhibitory effect is stronger than the inhibitory effect of at least one other screened test substance, and providing this selected test substance as an inhibitor of FKBP1A; (b) incubating cancer cells of a cancer cell line with the FKBP1A inhibitor and determining a property of the cancer cells; (c) incubating cancer cells of the same cancer cell line as in step (b) in the absence of the FKBP1A inhibitor and determining the same property of the cancer cells as in step (b) under the same conditions as step (b); (d) comparing the determined property of the cancer cells according to steps (b) and (c).
  6. The method according to any of the preceding claims, wherein the cancer cell line is characteristic of triple-negative breast cancer; preferably is a cancer cell line of triple-negative breast cancer; more preferably is a cancer cell line of triple-negative breast cancer of the mesenchymal stem-like subtype.
  7. The method according to any of the preceding claims, comprising the steps (a) providing an inhibitor (i) of FKBP1A expression; (b) incubating cancer cells of a cancer cell line of triple-negative breast cancer, preferably of the mesenchymal stem-like subtype, more preferably of the MDA-MB-231 cancer cell line; with the inhibitor of FKBP1A, and determining a property of the cancer cells, preferably the cellular differentiation status, more preferably by quantifying mesenchymal markers; (c) incubating cancer cells of the same cancer cell line as in step (b) in the absence of the FKBP1A inhibitor and determining the same property of the cancer cells as in step (b) under the same conditions as step (b); (d) comparing the determined property of the cancer cells according to steps (b) and (c).
  8. The method according to any one of claims 1 to 7, comprising the steps of (a) providing an inhibitor (i) of FKBP1A expression; (b) incubating cancer cells of a triple-negative breast cancer cell line, preferably of the mesenchymal stem-like subtype, more preferably of the MDA-MB-231 cancer cell line, with the FKBP1A inhibitor, and determining a property of the cancer cells, preferably cellular immunogenicity, more preferably by quantifying immunomodulatory proteins; (c) incubating cancer cells of the same cancer cell line as in step (b) in the absence of the FKBP1A inhibitor and determining the same property of the cancer cells as in step (b) under the same conditions as step (b); (d) comparing the determined property of the cancer cells according to steps (b) and (c).
  9. The method according to any one of claims 1 to 7, comprising the steps of (a) providing an inhibitor (i) of FKBP1A expression; (b) incubating cancer cells of a triple-negative breast cancer cell line, preferably of the mesenchymal stem-like subtype, more preferably of the MDA-MB-231 cancer cell line, with the FKBP1A inhibitor, and determining a property of the cancer cells, preferably cellular stem cell character, more preferably by quantifying (tumor) stem cell markers; (c) incubating cancer cells of the same cancer cell line as in step (b) in the absence of the FKBP1A inhibitor and determining the same property of the cancer cells as in step (b) under the same conditions as step (b); (d) comparing the determined property of the cancer cells according to steps (b) and (c).
  10. The method according to any one of claims 1 to 7, comprising the steps of (a) providing an inhibitor (i) of FKBP1A expression; (b) incubating cancer cells of a triple-negative breast cancer cell line, preferably of the mesenchymal stem-like subtype, more preferably of the MDA-MB-231 cancer cell line, with the FKBP1A inhibitor, and determining a property of the cancer cells, preferably quantifying the expression of FKBP1A; (c) incubating cancer cells of the same cancer cell line as in step (b) in the absence of the FKBP1A inhibitor and determining the same property of the cancer cells as in step (b) under the same conditions as step (b); (d) comparing the determined property of the cancer cells according to steps (b) and (c).
  11. The method according to any of the preceding claims, wherein step (a) comprises the sub-steps (a 1 ) providing a plurality of test substances; (a 2 ) screening the test substances for their inhibitory effect on (i) the expression of FKBP1A; (a 3 ) selecting at least one screened test substance whose inhibitory effect is stronger than the inhibitory effect of at least one other screened test substance, and providing this selected test substance as an inhibitor of FKBP1A.
  12. The method according to any of the preceding claims, wherein the triple-negative breast cancer corresponds to the mesenchymal stem-like subtype.
  13. An FKBP1A-specific siRNA for use in the treatment of triple-negative breast cancer; preferably wherein the triple-negative breast cancer corresponds to the mesenchymal stem-like subtype.
  14. (A) An inhibitor (i) of the expression of FKBP1A; and/or (ii) of the enzymatic activity of FKBP1A; and/or (iii) of the interaction(s) of FKBP1A with interaction partner(s); and/or (B) a PROTAC (PROteolysis TArgeting Chimeras) that specifically induces the cellular degradation of FKBP1A, namely FKBP12 PROTAC RC32 (CAS No.: 2375555-66-9), for use in the treatment of triple-negative breast cancer; wherein the inhibitor is a - FK506; and/or - a specific inhibitor of FKBP1A without immunosuppressive effect for synergistic immunotherapy, namely ElteN378, V-10367 (Vertex, USA), GPI-1046 (Guilford, USA), GPI-1485 (Guilford, USA), SLF, FK1706, FK-1012, AG5437 and/or AG5507.
  15. The inhibitor for use according to claim 14 for the adjuvant treatment of triple-negative breast cancer; preferably for (i) adjuvant treatment in immunotherapy of triple-negative breast cancer and/or for (ii) preventing metastasis formation and/or for (iii) targeted therapy of metastasis-inducing circulating tumor cells and/or cell clusters (CTC and/or CTC clusters), preferably with additional targeting of PIN1.

Description

Es werden die Prioritäten beansprucht von deutscher Patentanmeldung Nr. 10 2020 203 224.6, eingereicht am 12. März 2020, sowie von deutscher Patentanmeldung Nr. 10 2020 207 900.5, eingereicht am 25. Juni 2020. Die Erfindung betrifft ein Verfahren zum Auffinden von Inhibitoren der Peptidyl-prolyl cis-trans Isomerase FKBP1A oder von Antikörpern, Proteinen oder Molekülen mit spezifischer Affinität zu FKBP1A. Die Erfindung betrifft außerdem FKBP1A-spezifische siRNA, Inhibitoren der Expression von FKBP1A, Inhibitoren der enzymatischen Aktivität von FKBP1A, sowie Inhibitoren der Interaktion(en) von FKBP1A mit Interaktionspartner(n) jeweils zur Behandlung von Erkrankungen, insbesondere Krebserkrankungen oder neurodegenerative Erkrankungen. Darüber hinaus betrifft die Erfindung die Verwendung von FKBP1A als prognostischer oder diagnostischer Marker von Krebserkrankungen. Bei den Krebserkrankungen handelt es sich bevorzugt um das Mammakarzinom, insbesondere um das Triple-negative Mammakarzinom (TNBC, triple-negative breast cancer), ganz bevorzugt um den mesenchymal stem-like Subtyp. Die unabhängigen Ansprüche 1, 13 und 14 definieren die Erfindung. Die abhängigen Ansprüche definieren bevorzugte Ausführungsformen der Erfindung. Das Triple-negative Mammakarzinom wird bei 10-15% aller Brustkrebspatientinnen diagnostiziert und ist durch eine hohe Rezidivrate, aggressives Wachstum, frühe Metastasierung und schlechte Prognose charakterisiert. Kennzeichnend und wahrscheinlich mitursächlich für diese hohe Aggressivität sind der epigenetisch fehlgesteuerte geringe Differenzierungsstatus sowie der hohe Stammzellcharakter der Tumorzellen. Dies trifft insbesondere auf den mesenchymal stem-like Subtyp des TNBC zu. Im Gegensatz zu anderen Subformen des Mammakarzinoms wie z.B. des Hormonrezeptor- oder des Her2-positiven Mammakarzinoms besteht die aktuelle Therapie des Triple-negativen Mammakarzinoms gemäß den aktuellen Leitlinien ausschließlich in einer Behandlung mit Chemotherapeutika. Diese wirken jedoch unspezifisch mit bedeutenden Nebenwirkungen im gesunden Gewebe. Bis heute stehen beim Triple-negativen Mammakarzinom aufgrund der negativen Expression bekannter onkologischer Zielstrukturen (Östrogenrezeptor, Progesteronrezeptor oder Her2-Rezeptor) keine pharmakologisch adressierbaren zellulären Zielstrukturen für Therapieansätze zur Verfügung (vgl. Kumar, P. & Aggarwal, R. An overview of triple-negative breast cancer. Archives of Gynecology and Obstetrics (2016). doi:10.1007/s00404-015-3859-y). Viele zielgerichtete Therapieansätzen wurden bzw. werden in klinischen Studien untersucht, wie z.B. der Einsatz von PARP-Inhibitoren, Angiogenese-Inhibitoren oder mTOR-Inhibitoren, oder eine EGFR-gerichtete Behandlung. Diese Ansätze zeigten jedoch nur eine begrenzte therapeutische Effizienz (Fedele, P., Orlando, L. & Cinieri, S. Targeting triple negative breast cancer with histone deacetylase inhibitors. Expert Opinion on Investigational Drugs (2017). doi:10.1080/13543784.2017.1386172). Bis heute konnte keine effiziente personalisierte Therapie des Triple-negativen Mammakarzinoms entwickelt werden. Epigenetische Wirkstoffe, z.B. HDAC-Inhibitoren, befinden sich ebenfalls in ersten klinischen Studien zur Behandlung des Triple-negativen Mammakarzinoms. Dem Ansatz, mit epigenetischen Wirkstoffen den hohen Stammzellcharakter, die geringe Apoptoseinduktion sowie die geringe Immunogenität der Krebszellen zu adressieren, wird ein großes therapeutisches Potential zugesprochen (Fedele, P., Orlando, L. & Cinieri, S. Targeting triple negative breast cancer with histone deacetylase inhibitors. Expert Opinion on Investigational Drugs (2017). doi:10.1080/13543784.2017.1386172; Mazzone, R., Zwergel, C., Mai, A. & Valente, S. Epi-drugs in combination with immunotherapy: a new avenue to improve anticancer efficacy. Clinical Epigenetics (2017). doi:10.1186/s13148-017-0358-y). Die aktuell untersuchten Wirkstoffe zielen jedoch lediglich auf allgemeine zelluläre epigenetische Prozesse und sind wenig tumorspezifisch (Roberti, A., Valdes, A. F., Torrecillas, R., Fraga, M. F. & Fernandez, A. F. Epigenetics in cancer therapy and nanomedicine. Clinical Epigenetics (2019). doi:10.1186/s13148-019-0675-4). US 2009/0215812 betrifft Zusammensetzungen und Verfahren zum Bewerten der Wahrscheinlichkeit, dass ein Tumor gegenüber einem mTOR-Inhibitor, z. B. Rapamycin oder einem Rapamycin-Analogon, empfindlich ist. US 2016/0235731 betrifft bifunktionelle Verbindungen, die als Proteinabbau induzierende Einheiten wirken und Verfahren zum gezielten Abbau endogener Proteine durch Verwendung der bifunktionellen Verbindungen, die eine Cereblon-bindende Einheit an einen Liganden binden, der in der Lage ist, an das Zielprotein zu binden, das bei der Behandlung von proliferativen Störungen verwendet werden kann. US 2019/0092788 betrifft 32-Desoxo-Rapamycin-Derivate und deren Verwendungsverfahren. WO 2009/030770 betrifft Verfahren und Werkzeuge zum Erhalten einer effizienten Prognose (