Compositions targeting giant cells in blood disorders

Abstract

The subject matter disclosed herein relates to a method of treating a Blood Disorder in a subject in need thereof, the method comprising administering to the subject with a Blood Disorder a composition targeting a telomerase in a Giant Cell of the Disorder.

Description

[0001] COMPOSITIONS TARGETING GIANT CELLS IN BLOOD DISORDERS

[0002] CROSS-REFERENCE TO A RELATED APPLICATION

[0003] This application claims priority to U.S. Provisional Application No. 63 / 508,087, filed June 14, 2023 which is incorporated by reference for all purposes.

[0004] This patent disclosure contains material that is subject to copyright protection. The copyright owner has no objection to the facsimile reproduction by anyone of the patent document or the patent disclosure as it appears in the U.S. Patent and Trademark Office patent file or records, but otherwise reserves any and all copyright rights.

[0005] All patents, patent applications and publications cited herein are hereby incorporated by reference in their entirety. The disclosure of these publications in their entireties are hereby incorporated by reference into this application in order to more fully describe the state of the art as known to those skilled therein as of the date of the invention described herein.

[0006] BACKGROUND

[0007] The terms “Blood Disorders” and “Blood Disorder” here refer specifically to Myelodysplastic syndromes (MDS) and Acute myeloid leukemia (AML).

[0008] Myelodysplastic syndromes are a group of disorders caused by blood cells that are poorly formed or do not function properly. This syndrome can be caused by inherited changes in the Runx-related transcription factor 1 (RU NXI ) gene, which control the development of blood cells. Acute myeloid leukemia is a cancer of the blood and bone marrow. MDS and AML are both considered “Blood Disorders”. Myelodysplastic syndromes (MDS) and acute myeloid leukemia (AML) are incurable diseases except for <10% individuals who can be successfully treated with allogeneic transplantation (1, 2). Most therapies fail to produce sustained responses because of rapid clonal evolution and appearance of disease resistance (3, 4).

[0009] Telomerase is an enzyme involved in the maintenance of telomers which are repeated sequences that cap the ends of chromosomes.

[0010] CD 163 is a molecule that is primarily expressed on the surfaces of macrophages. It regulates inflammatory responses and has a role in iron metabolism and wound healing.

[0011] Giant cells are cells often formed by the fusion of multiple cells, including a macrophage / monocyte cell, which can be associated with tumors. Giant cells can range in size upward from 40 microns.

[0012] SUMMARY OF THE INVENTION

[0013] A first aspect of the invention is directed to a method of treating a Blood Disorder in a subject comprising administering to a subject in need thereof a drug or biological product that inhibits telomerase activity, or inhibits expression of, telomerase in a Giant Cell. The Blood Disorder may be myelodysplastic syndrome (MDS) or acute myeloid leukemia (AML).

[0014] A related aspect of the invention further comprises detecting or isolating a Giant Cell from the subject with a Blood Disorder prior to administering the drug or biological product, wherein the Giant Cell expresses telomerase.

[0015] A related aspect is directed to a composition for treating a Blood Disorder comprising a drug or biological product that inhibits telomerase activity, or inhibits expression of, telomerase in a Giant Cell. Such drugs or biological agents may comprise telomerase-specific antisense oligonucleotide, such as Imetelstat, and / or a small molecule inhibitor of telomerase such as BIRBI 52 or comprising antibody drug conjugates targeted to Giant Cells expressing telomerase.

[0016] Another aspect of the invention is directed to a method of treating a Blood Disorder in a subject comprising administering to a subject in need thereof a drug or biological product that binds to a macrophage marker on a Giant Cell and inhibits telomerase activity. The Blood Disorder includes myelodysplastic syndrome (MDS) or acute myeloid leukemia (AML).

[0017] A related aspect is directed to a composition for treating a Blood Disorder comprising a drug or biological product that binds to the macrophage marker CD 163. The composition may comprise a conjugate drug antibody such as a monoclonal antibody or other ligand that targets a macrophage marker on a Giant Cell and a second active component or moiety inhibiting telomerase activity.

[0018] Another aspect of the invention is directed to a combination of therapies or detection methods which target expression of telomerase on Giant Cells as well as expression of macrophage markers such as CD 163 on Giant Cells.

[0019] BRIEF DESCRIPTION OF FIGURES

[0020] This application fde contains at least one drawing executed in color. Copies of this patent application publication with color drawing(s) will be provided by the Office upon request and payment of the necessary fee.

[0021] FIGS. 1A and IB show Giant Cells from MDS patients showing CDI63 expression. Giemsa stained (left) and CD 163 stained (right) giant cell isolated from peripheral blood of a patient with MDS.

[0022] FIG. 1A shows Giemsa staining.

[0023] FIG. IB shows staining for CD163. FIGS. 2A-2D show that giant cells from MDS patients show telomerase expression.

[0024] Giant cells show strong positive signals for hTERT (FIGS. 2A, 2B and 2C) whereas small cells show weak signals for human telomerase reverse transcriptase (hTERT) (FIG. 2D).

[0025] FIG. 3 shows a schematic representation of CD 163 targeting-Imetelstat conjugate ADC.

[0026] DETAILED DESCRIPTION OF THE INVENTION

[0027] In certain aspects, the subject matter disclosed herein provides a method of treating a Blood Disorder in a subject in need thereof, the method comprising administering to the subject a composition targeting Giant Cells expressing telomerase. In some embodiments, the Blood Disorder is Myelodysplastic syndromes (MDS). In some embodiments, the Blood Disorder is acute myeloid leukemia (AML).

[0028] As mentioned above, myelodysplastic syndromes (MDS) and acute myeloid leukemia (AML) are incurable diseases except for < 10% individuals who can be successfiilly treated with allogeneic transplantation. Most therapies fail to produce sustained responses because of rapid clonal evolution and appearance of disease resistance.

[0029] In view of the failure and limitations of past treatments, the inventors searched for the presence of circulating giant cells (GCs) in patients with MDS using a highly sensitive sizebased method (ISET) commonly used to isolate circulating tumor cells.

[0030] GC cells have a size of 40 microns or larger. The inventors found GC in 46 / 99 MDS tested patients and in 1 1 / 66 healthy controls through the analysis of, on average, 9 ml of blood. Overall, 408 GC have been identified in MDS patients and 28 GC in controls. GC could be characterized using immunomorphological analyses. 69.4% of GC in MDS patients scored positive for macrophage specific antibodies (CD163, CDl lb, CD68,) (FIGS. 1A-1B). Interestingly it was also found that 74.4% of GC were strongly positive with the telomerase specific antibody (FIGS. 2A-D). Furthermore, 97.4% of GC scored positive for CD61 and 54% of GC were positive for Runx2. These results show for the first time that the detection of GC in the peripheral blood of MDS patients is not a rare event and demonstrate the clinical relevance of GC pathogenesis in patients with MDS.

[0031] The inventors considered several approaches to targeting telomerase function. Some direct approaches involve targeting the telomerase itself with oligonucleotide inhibitors such as Imetelstat (lipidated 13-mer thiophosphoramidate oligonucleotide complementary to the TERC template region) in AML or MDS. To achieve specific targeting, the inventors consider a conjugate drug compound comprising Imetelstat or other telomerase-specific antisense oligonucleotides (11) with antibodies binding to CD 163 or other macrophage markers.

[0032] FIG. 3 shows a schematic representation of CD 163 targeting-Imetelstat conjugate ADC in Blood Disorders. A small molecule inhibitor, BIBRI 532 (non-competitive small molecule inhibitor of telomerase), can also target telomerase in Blood Disorders.

[0033] Since GC are characterized by the presence of macrophage markers, including CD 163, and high telomerase expression, this offers a unique opportunity to specifically target GC and spare normal macrophages that show' no or low' telomerase activity. Except for B / plasma cells, the majority of the terminally differentiated cell types in the body do not show7telomerase expression.

[0034] Embodiments of the invention include the following:

[0035] This disclosure provides a method of treating a Blood Disorder in a subject in need thereof, the method comprising administering to the subject a composition targeting telomerase expression in a Giant Cell of the Blood Disorder expressing CD 163; wherein the Blood Disorder is Myelodysplastic syndrome (MDS) or acute myeloid leukemia (AML).

[0036] In one embodiment the subject is in need of treatment of MDS. In another embodiment of this method, the subject is in need of treatment for AML. In some embodiments, the composition comprises a telomerase-specific antisense oligonucleotide. In some embodiments, the telomerase-specific antisense oligonucleotide is Imetelstat.

[0037] In some embodiments, the composition comprises a molecule specific for an antigen on the Giant Cell, wherein the antigen comprises a macrophage marker, and wherein the molecule is conjugated to a telomerase inhibitor.

[0038] In some embodiments, the macrophage marker is CD 163. In some embodiments, the telomerase inhibitor is a telomerase-specific antisense oligonucleotide.

[0039] In some embodiments, the telomerase-specific antisense oligonucleotide is Imetelstat. In some embodiments, the telomerase inhibitor is a small molecule inhibitor, BIBRI 532.

[0040] In some embodiments, the molecule is an antibody specific for a macrophage marker.

[0041] In some embodiments, the molecule is an antibody specific for CD 163. The antibody may be a monoclonal antibody or an antigen-binding fragment of an antibody.

[0042] In some embodiments, the composition comprises a small molecule inhibitor of the telomerase such as, but not limited to, BIBRI532.

[0043] Other embodiments of the invention include but are not limited to those which follow.

[0044] 1 . A method of treating a Blood Disorder in a subject comprising administering to a subject in need thereof a drug or biological product that inhibits telomerase activity, or inhibits expression of telomerase in a Giant Cell.

[0045] 2. The method of embodiment 1 , wherein the Giant Cell from the subject expresses telomerase

[0046] 3. The method of embodiment 1 , wherein the Blood Disorder is myelodysplastic syndrome (MDS).

[0047] 4. The method of embodiment 1 , wherein the Blood Disorder is acute myeloid leukemia

[0048] (AML). The method of embodiment 1 , wherein the drug or biological product comprises a telomerase-specific antisense oligonucleotide. The method of embodiment 1 , wherein the telomerase-specific antisense oligonucleotide is Imetelstat. The method of embodiment 1 , wherein the drug or biological product comprises a small molecule inhibitor of telomerase. The method of embodiment 1 , wherein the drug or biological product comprises BIRB1532. The method of embodiment 1 , wherein the drug or biological product comprises an antibody or antibody conjugate that comprises a telomerase-specific antisense oligonucleotide such as Imetelstat and / or a small molecule inhibitor of telomerase such as BIRB1532. The method of any one of embodiments 1-9, further comprising detecting or isolating a Giant Cell from the subject with MDS or AML prior to administering the drug or biological product; wherein the Giant Cell expresses telomerase. The method of embodiment 10, wherein the Giant Cell is detected or isolated from the blood of the subject with MDS or AML A composition for treating a Blood Disorder comprising a drug or biological product that inhibits telomerase activity, or inhibits expression of telomerase in a Giant Cell. The composition of embodiment 12 that comprises a telomerase-specific antisense oligonucleotide, such as Imetelstat, and / or a small molecule inhibitor of telomerase such as BIRB152 The composition of embodiment 12 that comprises an antibody conjugate comprising an antibody that binds to CD 163 on a Giant Cell and a telomerase-specific antisense oligonucleotide, such as Imetelstat, and / or a small molecule inhibitor of telomerase such as BIRBI 52. The composition of embodiment 14 that comprises a monoclonal antibody that binds to a Giant Cell from a MDS or AML patient expressing telomerase. A method of treating a Blood Disorder in a subject comprising administering to a subject in need thereof a drug or biological product that binds to a macrophage marker in or on a Giant Cell. The method of embodiment 16, wherein the Blood Disorder is myelodysplastic syndrome (MDS). The method of embodiment 16, wherein the Blood Disorder is acute myeloid leukemia (AML). The method of embodiment 16, wherein the composition comprises an antibody or other ligand that binds to the macrophage marker. The method of any one of embodiments 16-19, further comprising detecting or isolating a Giant Cell from the subject with MDS or AML prior to administering the drug or biological product; wherein the Giant Cell expresses a macrophage marker and telomerase. The method of embodiment 20, wherein the Giant Cell is detected or isolated from the blood of the subject with MDS or AML. The method of embodiment 20, wherein the drug or biological product is an antibody that binds to a macrophage marker on the Giant Cell. The method embodiment 20, wherein the macrophage marker on the Giant Cell is

[0049] CD163. 24. The method of embodiment 20 that comprises use of an antibody conjugate comprising an antibody that binds to the macrophage marker CD 163 on a Giant Cell and the drug inhibitor of telomerase.

[0050] Other embodiments are described by the following claims.

[0051] References:

[0052] 1. Jam AG, Elmariah H. BMT for Myelodysplastic Syndrome: When and Where and How. Front Oncol. 2021; 1 1 :771614.

[0053] 2. Litzow MR, Tarima S, Perez WS, Bolwell BJ, Cairo MS, Camitta BM, et al. Allogeneic transplantation for therapy-related myelodysplastic syndrome and acute myeloid leukemia. Blood. 201O;l 15(9): 1850-7.

[0054] 3. da Silva-Coelho P, Kroeze LI, Yoshida K, Koorenhof-Scheele TN, Knops R, van de Locht LT, et al. Clonal evolution in myelodysplastic syndromes. Nat Commun. 2017;8: 15099.

[0055] 4. Makishima H, Yoshizato T, Yoshida K, Sekeres MA, Radivoyevitch T, Suzuki H, et al. Dynamics of clonal evolution in myelodysplastic syndromes. Nat Genet.

[0056] 20I7;49(2):204-12.

[0057] 5. Adams DL, Martin SS, Alpaugh RK, Charpentier M, Tsai S, Bergan RC, et al. Circulating giant macrophages as a potential biomarker of solid tumors. Proc Natl Acad Sci US A 2014;l l l(9):3514-9.

[0058] 6. Mirzayans R, Andrais B, Murray D. Roles of Polyploid / Multinucleated Giant Cancer Cells in Metastasis and Disease Relapse Following Anticancer Treatment. Cancers (Basel).20I8; 10(4).

[0059] 7. Mirzayans R, Andrais B, Scott A, Wang YW, Kumar P, Murray D. Multinucleated Giant Cancer Cells Produced in Response to Ionizing Radiation Retain Viability and Replicate Their Genome. Int J Mol Sci. 2017; 18(2).

[0060] 8. Ohashi R, Hayama A, Matsubara M, Watarai Y, Sakatani T, Naito Z, et al. Breast carcinoma with osteociast-like giant cells: A cytological-pathological correlation with a literature review. AnnDiagnPathol. 2018;33:1-5.

[0061] 9. Pienta KJ, Hammarlund EU, Axelrod R, Brown JS, Amend SR. Poly-aneuploid cancer cells promote evolvability, generating lethal cancer. Evol Appl. 2020; 13(7): 1626-34. 10. Pienta KJ, Hammarlund EU, Brown JS, Amend SR, Axelrod RM. Cancer recurrence and lethality are enabled by enhanced survival and reversible cell cycle arrest of polyaneuploid cells. Proc Natl Acad Sci U S A 2021 ; 118(7).

[0062] 11. Vonderheide RH. Telomerase as a universal tumor-associated antigen for cancer immunotherapy. Oncogene. 2002;21 (4):674-9.

Claims

CLAIMSWhat is claimed is:

1. A method of treating a Blood Disorder in a subject in need thereof, the method comprising administering to the subject a composition targeting a telomerase in a Giant Cell of the blood disorder.

2. The method of claim 1, wherein the blood disorder is Myelodysplastic syndromes (MDS).

3. The method of claim 1, wherein the blood disorder is acute myeloid leukemia (AML).

4. The method of claim 1, wherein the composition comprises a telomerase-specific antisense oligonucleotide.

5. The method of claim 4, wherein the telomerase-specific antisense oligonucleotide is Imetelstat.

6. The method of claim 1, wherein the composition comprises a molecule specific for an antigen on the Giant Cell, wherein the antigen comprises a macrophage marker, and wherein the molecule is conjugated to a telomerase inhibitor. The method of claim 6, wherein the macrophage marker is CD 163.

7. The method of claim 6, wherein the telomerase inhibitor is a telomerase-specific antisense oligonucleotide.

8. The method of claim 7, wherein the telomerase-specific antisense oligonucleotide is Imetelstat.

9. The method of claim 6, wherein the telomerase inhibitor is a small molecule inhibitor.

10. The method of claim 9, wherein the small molecule inhibitor is BIBRI 532.

11. The method of claim 6, wherein the molecule is an antibody specific for a macrophage marker.

12. The method of claim 6, wherein the molecule is an antibody specific for CD 163.

13. The method of claim 1, wherein the composition comprises a small molecule inhibitor of the telomerase.

14. The method of claim 13, wherein the small molecule inhibitor is BIBRI 532.

15. The method of any one of claims 1-14, further comprising detecting or isolating a Giant Cell from the subject with Blood Disorder prior to administering a composition; wherein the Giant Cell expresses telomerase.

16. The method of claim 15, wherein the composition comprises a telomerase-specific antisense oligonucleotide.

17. The method of claim 16, wherein the telomerase-specific antisense oligonucleotide is Imetelstat.

18. The method of claim 15, wherein the composition comprises a molecule specific for an antigen on the Giant Cell, wherein the antigen comprises a macrophage marker, and wherein the molecule is conjugated to a telomerase inhibitor.

19. The method of claim 18, wherein the macrophage marker is CD163.

20. The method of claim 18, wherein the telomerase inhibitor is a telomerase-specific antisense oligonucleotide.

21. The method of claim 20, wherein the telomerase-specific antisense oligonucleotide is Imetelstat.

22. The method of claim 18, wherein the telomerase inhibitor is a small molecule inhibitor.

23. The method of claim 22, wherein the small molecule inhibitor is BIBRI 532.

24. The method of claim 18, wherein the molecule is an antibody specific for a macrophage marker.

25. The method of claim 18, wherein the molecule is an antibody specific for CD 163.

26. The method of claim 15, wherein the composition comprises a small molecule inhibitor of the telomerase.

27. The method of claim 26, wherein the small molecule inhibitor is BIBRI 532.

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