Combinations for treating cancer

Abstract

The present invention relates to combinations, pharmaceutical compositions, and kits comprising tinostamustine and an immune checkpoint inhibitor. These combinations, pharmaceutical compositions, and kits are effective for use in the treatment of cancer.

Description

[Technical Field]

[0001] The present invention relates to combinations comprising tinostamustine, or a pharmaceutically acceptable salt thereof, and an immune checkpoint inhibitor. The present invention further relates to the use of such combinations in the treatment of cancer, in particular to the medical use of the combinations described herein. [Background technology]

[0002] Manipulating the immune system by blocking ligands and receptors that act as regulators of the immune response (so-called immune checkpoints) has become an important strategy in cancer treatment in recent years. Therapies that block ligands and receptors that act as regulators of the immune response are sometimes referred to as "immune checkpoint inhibitors" in this technology. To date, several immune checkpoint inhibitors have been approved for clinical use in various cancers. Examples of immune checkpoint inhibitors include cytotoxic T lymphocyte-associated antigen 4 (CTLA-4) inhibitors and programmed cell death protein 1 (PD-1) inhibitors, as well as inhibitors of its ligand (PD-L1).

[0003] Programmed cell death protein-1 (PD-1, CD279), a 55kD type I transmembrane protein, is a member of the CD28 family of T cell costimulatory receptors, which also includes CD28, CTLA-4, ICOS, and BTLA. PD-1 contains an intracellular membrane proximal immunoreceptor tyrosine repression motif (ITIM) and a membrane distal immunoreceptor tyrosine-based switch motif (ITSM). PD-1 is composed of four polypeptide chains, each containing two identical heavy chains and two identical light chains. PD-1 is a negative regulatory molecule primarily expressed on activated T cells, B cells, and myeloid cells (Nishimura, H. and T. Honjo, PD-1: an inhibitory immunoreceptor involved in peripheral tolerance. Trends Immunol, 2001. 22(5): pp. 265-268). PD-1 delivers a negative signal by recruiting the protein tyrosine phosphatase SHP-2 to phosphorylated tyrosine residues in the ITSM within its cytoplasmic domain (Sheppard, KA et al., PD-1 inhibits T-cell receptor induced phosphorylation of the ZAP70 / CD3zeta signalosome and downstream signaling to PKCtheta. FEBS Lett, 2004. 574(1-3): pp. 37-41).

[0004] Two ligands specific to PD-1, namely PD-L1 (B7-H1 / CD274) and PD-L2 (B7-DC / CD273), have been identified. Both PD-L1 and PD-L2 have been shown to downregulate T cell activation upon binding to PD-1 in both mouse and human systems (Latchman, Y. et al., PD-L2 is a second ligand for PD-1 and inhibits T cell activation. Nat Immunol, 2001. 2(3): pp. 261-268; Carter, L. et al., PD-1:PD-L inhibitory pathway affects both CD4(+) and CD8(+) T cells and is overcome by IL-2. Eur J Immunol, 2002. 32(3): pp. 634-643).

[0005] Nivolumab is a human monoclonal antibody (immunoglobulin G4[IgG4]-S228P) that targets programmed death-1 (PD-1). PD-1 blockade with nivolumab is considered a promising immunotherapy, and nivolumab has received clinical approval for several cancer types, including advanced NSCLC, melanoma, renal cell carcinoma, high-MSI colorectal cancer, squamous cell carcinoma of the head and neck, and Hodgkin's disease. When administered as a first-line treatment according to the CheckMate 066 trial, nivolumab has shown improved overall survival compared to dacarbazine chemotherapy in patients with advanced melanoma (Robert, C. et al., Nivolumab in previously untreated melanoma without BRAF mutation. N Engl J Med, 2015. 372(4): pp. 320-330). In CheckMate 066, overall survival improved by 58% (HR=0.42, p<0.001), while the objective response rate improved from 13.9% with dacarbazine to 40.0% with nivolumab, and progression-free survival improved by 57% from 2.2 months to 5.1 months (HR=0.43, p<0.001).

[0006] Nivolumab is approved for use in combination with the anti-CTLA-4 monoclonal antibody ipilimumab in patients with advanced melanoma, based on the results of the CheckMate 067 study, which showed improved median overall survival compared to nivolumab monotherapy (37.6 months vs. not achieved, HR=0.85, (95% CI, 0.68~1.07)) (Wolchok, JD et al., Overall Survival with Combined Nivolumab and Ipilimumab in Advanced Melanoma. N Engl J Med, 2017. 377(14): pp. 1345~1356). However, treatment of patients with the nivolumab and ipilimumab combination resulted in grade 3 or 4 serious treatment-related adverse events in 59% of patients. As a result, administration of the nivolumab and ipilimumab combination is limited to relatively younger patients with higher performance scores.

[0007] These data suggest that current standard treatments, including immune checkpoint inhibitors as monotherapy or in combination with other immune checkpoint inhibitors, can lead to adverse patient outcomes. Therefore, improved immunotherapy is needed to treat cancer.

[0008] Melanoma is a particularly difficult type of cancer to treat. Although melanoma accounts for only 1% of all malignant lesions, it is difficult to treat due to its high metastatic potential and mortality rate. In recent decades, the number of melanoma cases has increased more rapidly and dramatically than any other type of cancer. Patients diagnosed early have a 5-year survival rate of approximately 90%, but this figure drops to 10% with a median survival time of 6 to 12 months in patients with advanced melanoma. Thus, advanced melanoma has traditionally been associated with limited treatment options and a poor prognosis. Historically, the median survival time has ranged from 6 to 9 months, but in recent years, the emergence of new therapeutic agents (e.g., immune checkpoint inhibitors) has improved the treatment of advanced melanoma. Objective responses to immune checkpoint inhibitors can be observed in more than 50% of patients with advanced melanoma, and the survival rate after treatment has increased from 12 months to 48-60 months (Hodi FS, KH, Sznol M, Carvajal R, Lawrence D, Atkins M et al., Durable, long-term survival in previously treated patients with advanced melanoma (MEL) who received nivolumab (NIVO) monotherapy in a phase I trial. Cancer Res., 2016. 76). Nevertheless, approximately 9,000 patients still die from melanoma each year in the United States, highlighting the need for more effective treatments.

[0009] International Publication No. 2010 / 085377 discloses tinostamustin (also known as EDO-S101), a first-in-class alkylated deacetylase inhibitor (HDACi) molecule. Tinostamustin has shown anticancer activity in in vitro and in vivo models and is the subject of clinical trials for the treatment of glioblastoma (NCT05432375, NCT03452930), relapsed / refractory hematological malignancies (NCT02576496), and solid tumors (NCT03345485). [Prior art documents] [Chartered documents]

[0010]

Patent Document 1

Patent document 2

Patent Document 3

Patent document 4

Patent document 5

Patent document 6

Patent document 7

Patent document 8

Non-licensed literature

[0011] [Non-licensed document 1] Nishimura, H. and T. Honjo, PD-1: an inhibitory immunoreceptor involved in peripheral tolerance. Trends Immunol, 2001. 22(5): 265~8 pages [Non-licensed document 2] Sheppard, KAら, PD-1 inhibits T-cell receptor phosphorylation of the ZAP70 / CD3zeta signalosome and downstream signaling to PKCtheta. FEBS Lett, 2004. 574(1-3): pp. 37~41 [Non-licensed document 3] Latchman, Y. et al., PD-L2 is a second ligand for PD-1 and inhibits T cell activation. Nat Immunol, 2001. 2(3): pp. 261-8. [Non-Patent Document 4] Carter, L. et al., PD-1:PD-L inhibitory pathway affects both CD4(+) and CD8(+) T cells and is overcome by IL-2. Eur J Immunol, 2002. 32(3): pp. 634-43 [Non-Patent Document 5] Robert, C. et al. Nivolumab in previously untreated melanoma without BRAF mutation. N Engl J Med, 2015. 372(4): pp. 320-30 [Non-Patent Document 6] Wolchok, JD et al. Overall survival with Combined Nivolumab and Ipilimumab in Advanced Melanoma. N Engl J Med, 2017. 377(14): pp. 1345-1356 [Non-Patent Document 7] Hodi FS, KH, Sznol M, Carvajal R, Lawrence D, Atkins M, et al. Durable, long-term survival in previously treated patients with advanced melanoma (MEL) who received nivolumab (NIVO) monotherapy in a phase I trial. Cancer Res., 2016. 76 [Non-Patent Document 8] Kim SK, Cho SW. The Evasion Mechanisms of Cancer Immunity and Drug Intervention in the Tumor Microenvironment. Front Pharmacol. 2022 May 24;13:868695. doi: 10.3389 / fphar.2022.868695. PMID: 35685630; PMCID: PMC9171538 [Non-licensed Document 9] Han Y, Liu D, Li L. PD-1 / PD-L1 pathway: current researches in cancer. Am J Cancer Res. 2020 Mar 1;10(3):pages 727~742. PMID: 32266087; PMCID: PMC7136921 [Non-licensed Document 10] Johnson, Immune-checkpoint inhibitors: long-term implications of toxicity. Nat Rev Clin Oncol 19, pp. 254~267 (2022). https: / / doi.org / 10.1038 / s41571-022-00600-w [Non-licensed Document 11] EW Martin, "Remington's Pharmaceutical Sciences" [Non-licensed Document 12] Pitha, J Pharm Sci, 84 (8), pp. 927-32 (1995) [Non-licensed Document 13] Keilholzら(doi: https: / / doi.org / 10.1016 / j.annonc.2020.07.004) [Non-licensed Document 14] Van Wilpe S, Koornstra R, Den Brok M, De Groot JW, Blank C, De Vries J, Gerritsen W, Mehra N. Lactate dehydrogenase: a marker of diminished antitumor immunity. Oncoimmunology. 2020 Feb 26;9(1):1731942. doi: 10.1080 / 2162402X.2020.1731942. PMID: 32158624; PMCID: PMC7051189 [Non-Patent Document 15] Dubois D, Dubois EF, A formula to estimate the approximate surface area if height and weight be known, Arch Intern Med, 1916, 17, pp. 863-871. [Non-Patent Document 16] NCT01721772; Topalian et al., N Engl J Med 366: 2443~2454, 2012 [Non-Patent Document 17] Zimmer L et al; Ipilimumab alone or in combination with nivolumab after progression on anti-PD-1 therapy in advanced melanoma. Eur J Cancer. 2017 Apr;75:47~55. doi: 10.1016 / j.ejca.2017.01.009. Epub 2017 Feb 17. PMID: 28214657 [Non-Patent Document 18] Baron K, et al. [Overview of the project] [Problems that the invention aims to solve]

[0012] The object of the present invention is to address one or more of the above-mentioned problems. [Means for solving the problem]

[0013] This invention relates to the surprising and unexpected finding that when immune checkpoint inhibitors are administered in combination with the alkylated HDACi molecule, tinostamustine, the efficacy of immune checkpoint inhibitors in treating cancer may be enhanced. While we do not wish to be bound by theory, it is believed that tinostamustine enhances the efficacy of immune checkpoint inhibitors, and therefore results in improved overall progression-free survival and improved overall survival, as demonstrated in the clinical data described herein. The clinical data described herein demonstrate that the combination according to the present invention increases median progression-free survival (mPFS) and median overall survival (mOS) by several months compared to the current standard of care for advanced melanoma. This represents a significant increase in survival for patients for whom surgery is not an option and other treatment courses, including chemotherapy and radiotherapy, are considered ineffective or potentially ineffective. Given that immune checkpoint inhibitors are used in the treatment of a wide range of cancers, in addition to advanced melanoma, the unexpected discovery that the combination of this invention is effective in the treatment of advanced cancer is likely to have broader applicability in the treatment of cancer in general.

[0014] Furthermore, the combinations described herein have been found to avoid adverse events associated with conventional combinations of two or more immune checkpoint inhibitors used to treat cancer (e.g., the combination of nivolumab and ipilimumab). Therefore, the present invention not only addresses combinations that improve patient prognosis (as determined by mPFS and mOS) but also mitigates adverse side effects associated with immunotherapy.

[0015] According to a first aspect of the present invention, a combination comprising tinostamustine, or a pharmaceutically acceptable salt thereof, and an immune checkpoint inhibitor is provided.

[0016] The immune checkpoint inhibitor may be a programmed cell death protein-1 (PD-1) inhibitor. The immune checkpoint inhibitor may be an anti-PD-1 antibody. The immune checkpoint inhibitor may be selected from pembrolizumab, nivolumab, dostallimab, retifanlimab, and cemiprimab. The immune checkpoint inhibitor may be nivolumab.

[0017] The immune checkpoint inhibitor may be a programmed cell death protein ligand 1 (PD-L1) inhibitor and / or a programmed cell death protein ligand 2 (PD-L2) inhibitor. The immune checkpoint inhibitor may be a PD-L1 inhibitor. The immune checkpoint inhibitor may be an anti-PD-L1 antibody. The immune checkpoint inhibitor may be selected from atezolizumab, avelumab, and durvalumab.

[0018] The combination may further include one or more additional immune checkpoint inhibitors. One or more additional immune checkpoint inhibitors may include cytotoxic T lymphocyte-associated antigen 4 (CTLA-4) inhibitors. The CTLA-4 inhibitor may be ipilimumab. The CTLA-4 inhibitor may be tremelimumab.

[0019] A second aspect of the present invention provides a pharmaceutical composition comprising the combination according to the first aspect of the present invention and a pharmaceutically acceptable diluent or carrier.

[0020] According to a third aspect of the present invention, a kit is provided which includes a combination according to the first aspect of the present invention or a pharmaceutical composition according to the second aspect of the present invention, and optionally, instructions for treating a patient.

[0021] Embodiments provide a combination according to a first aspect of the present invention for therapeutic use (for example, for pharmaceutical use), a pharmaceutical composition according to a second aspect of the present invention, or a kit according to a third aspect of the present invention.

[0022] According to a fourth aspect of the present invention, a combination according to the first aspect of the present invention, a pharmaceutical composition according to the second aspect of the present invention, or a kit according to the third aspect of the present invention is provided for use in the treatment of cancer. Cancers may be selected from melanoma, small cell lung cancer, non-small cell lung cancer, mesothelioma (e.g., malignant pleural mesothelioma), renal cell carcinoma, head and neck cancer, urothelial carcinoma, colorectal cancer, esophageal squamous cell carcinoma, hepatocellular carcinoma, gastric cancer, esophageal cancer, esophageal adenocarcinoma, gastroesophageal junction cancer, high microsatellite instability or mismatch repair deficiency cancer, primary mediastinal large B-cell lymphoma (PMBCL), Merkel cell carcinoma (MCC), endometrial cancer, high tumor mutational load (TMB-H) cancer, cutaneous squamous cell carcinoma (cSCC), soft tissue sarcoma, osteosarcoma, ovarian cancer, trinegative breast cancer, brain cancer (e.g., glioblastoma), multiple myeloma, Hodgkin lymphoma, and non-Hodgkin lymphoma. Cancers may be melanoma, or, at the discretion of the researchers, advanced melanoma. Cancers may be recurrent and / or refractory.

[0023] In the treatment of cancer according to a fourth aspect of the present invention, tinostamustine, or a pharmaceutically acceptable salt thereof, may be administered to patients in need as an adjuvant and / or neoadjuvant, with the option of administering tinostamustine as a neoadjuvant to patients in need.

[0024] In a cancer treatment according to a fourth aspect of the present invention, tinostamustine, or a pharmaceutically acceptable salt thereof, is administered to the patient at a dose of 10-50 mg / m² of body surface area. 2 Optionally, 20-40 mg / m² 2 Additionally, an optional 25-35 mg / m² is available. 2 Furthermore, an optional 30 mg / m² is available. 2 It may be administered within the specified dosage range.

[0025] In the treatment of cancer according to a fourth aspect of the present invention, tinostamustine, or a pharmaceutically acceptable salt thereof, and an immune checkpoint inhibitor may be administered to the patient sequentially, simultaneously, or separately. Tinostamustine, or a pharmaceutically acceptable salt thereof, and an immune checkpoint inhibitor may be administered separately, and optionally, the immune checkpoint inhibitor may be administered first to the patient in need, followed by tinostamustine. For example, tinostamustine or a pharmaceutically acceptable salt thereof may be administered to the patient in need 20 to 120 minutes after administration of the immune checkpoint inhibitor to the patient, optionally 30 to 90 minutes later, and optionally a further 50 to 70 minutes (e.g., 60 minutes) later. Preferably, tinostamustine or a pharmaceutically acceptable salt thereof may be administered to the patient in need at least 30 minutes after administration of the immune checkpoint inhibitor to the patient.

[0026] In a cancer treatment according to a fourth aspect of the present invention, in the first cycle of treatment, tinostamustine or a pharmaceutically acceptable salt thereof may be administered to the patient in need without a checkpoint inhibitor, and in all subsequent cycles (e.g., the second, third, and fourth cycles), tinostamustine and / or a pharmaceutically acceptable salt thereof and an immune checkpoint inhibitor may be administered to the patient successively, simultaneously, or separately, preferably separately.

[0027] According to a fifth aspect of the present invention, tinostamustine, or a pharmaceutically acceptable salt thereof, is provided for use in the treatment of cancer in combination with an immune checkpoint inhibitor. Cancer and / or the treatment may be as defined by a fourth aspect of the present invention.

[0028] According to a sixth aspect of the present invention, an immune checkpoint inhibitor is provided for use in the treatment of cancer, wherein the immune checkpoint inhibitor is used in combination with tinostamstine or a pharmaceutically acceptable salt thereof. Cancer and / or said treatment may be as defined by a fourth aspect of the present invention.

[0029] A seventh aspect of the present invention provides a method for treating cancer, comprising administering to a patient in need a therapeutically effective dose of tinostamustine, or a pharmaceutically acceptable salt thereof, in combination with a therapeutically effective dose of an immune checkpoint inhibitor. The cancer and / or the treatment may be as defined by the fourth aspect of the present invention.

[0030] According to an eighth aspect of the present invention, the use of a combination comprising tinostamustine, or a pharmaceutically acceptable salt thereof, and an immune checkpoint inhibitor in the treatment of cancer is provided. Cancer and / or said treatment may be as defined by a fourth aspect of the present invention.

[0031] According to a ninth aspect of the present invention, the use of tinostamustine or a pharmaceutically acceptable salt thereof in the manufacture of a pharmacopoeia for use in the treatment of cancer is provided, wherein the tinostamustine or a pharmaceutically acceptable salt thereof is used in combination with an immune checkpoint inhibitor in the treatment. Cancer and / or the treatment may be as defined by a fourth aspect of the present invention.

[0032] According to a tenth aspect of the present invention, the use of an immune checkpoint inhibitor in the manufacture of a pharmacopoeia for use in the treatment of cancer is provided, wherein the immune checkpoint inhibitor is used in combination with tinostamstine or a pharmaceutically acceptable salt thereof in the treatment. Cancer and / or the treatment may be as defined by a fourth aspect of the present invention.

[0033] According to an eleventh aspect of the present invention, tinostamustine or a pharmaceutically acceptable salt thereof is provided for use in the treatment of melanoma, optionally for use in the treatment of progressive melanoma, and optionally for use in the treatment of recurrent and / or refractory melanoma.

[0034] According to a twelfth aspect of the present invention, tinostamustine or a pharmaceutically acceptable salt thereof is provided for use in the treatment of colorectal cancer, in which, optionally, the colorectal cancer is a carcinoma, and again optionally, the carcinoma is an adenocarcinoma. [Brief explanation of the drawing]

[0035] [Figure 1] This graph shows progression-free survival (PFS) in patients treated with the combination of tinostamustine and nivolumab according to the examples described herein, and a comparison of median progression-free survival (mPFS) versus the standard treatment reference (Zimmer et al.). “All mPFS” refers to the mPFS calculated for all patients mobilized for the study, as described in the examples herein. “mPFS Responders” refers to the mPFS calculated for patients considered to respond to the treatment, as described in the examples herein. “mPFS ipi / nivo(Zimmer)” refers to the reference mPFS value for patients treated with the combination of ipilimumab and nivolumab, as provided by Zimmer et al., as described in the examples herein. The patient reference number corresponds to the number assigned to each patient in Table 1 of the examples herein. [Figure 2] This graph shows overall survival (OS) and median overall survival (mOS) versus standard treatment reference (Baron et al.) in patients treated with the combination of tinostamustine and nivolumab according to the examples described herein. "All mOS" refers to the mOS calculated for all patients mobilized for the study, as described in the examples described herein. "mOS Responders" refers to the mOS calculated for patients considered to respond to the treatment, as described in the examples described herein. "mOS ipi / nivo (Baron)" refers to the reference mOS value in patients treated with the combination of ipilimumab and nivolumab, as provided by Baron et al., as described in the examples described herein. The patient reference number corresponds to the number assigned to each patient in Table 1 of the examples described herein. [Figure 3]This graph shows that tinostamustine as a monotherapy or in combination with an anti-PD1 antibody is well-tolerated and leads to weight gain in the MC38 colorectal cancer model. The graph also shows the mean body weight of mice in different groups during treatment in the MC38 model. [Figure 4] This graph shows that tinostamustine, as a combination therapy with an anti-PD1 antibody in an MC38 colorectal cancer model, significantly reduces tumor volume compared to tinostamustine as a monotherapy and to a control. P-values, Ns = not significant; P>0.05; ***: P<0.001. [Figure 5] This table shows that tinostamustine as a monotherapy or in combination with an anti-PD1 antibody has antitumor effects in the treatment of MC38. a. Mean ± SEM (number of mice); b. TGI% = [1-T / C] × 100%; RTV_TGI% = [1-RTV_T / RTV_C] × 100%; c. Bartlett's test was performed to test for uniformity of variance and normality, p < 0.05; then Kruskal-Wallis test was performed, p < 0.05; then Conover's nonparametric many-to-one comparison test was performed for each treatment against one control. ns: not significant; ***: P < 0.001. [Figure 6] This table shows that tinostamustine as a single agent, or as a combination treatment with an anti-PD1 antibody, increases the survival time of mice with colorectal cancer (MC38 model). Since more than 50% of mice survived at the end of the study in groups 2 through 6, median survival was not simply defined in these groups. Survival time was assessed by the time a mouse was found dead or euthanized. [Modes for carrying out the invention]

[0036] definition As used herein, the term “immune checkpoint inhibitor” means, unless otherwise specified, a drug molecule that can bind to an immune checkpoint receptor on the surface of an immune cell, thereby blocking the receptor and preventing the interaction between the receptor and its endogenous ligand. Alternatively, the term “immune checkpoint inhibitor” may mean a drug molecule that can bind to a ligand for an immune checkpoint receptor, thereby blocking the ligand from interacting with the immune checkpoint receptor. Immune checkpoint receptors are proteins typically presented on the surface of immune cells (e.g., T cells) involved in the checkpoint regulation of the immune response. Ligands for immune checkpoint receptors are also proteins and may be present, for example, on the surface of cancer cells.

[0037] More specifically, immune checkpoint inhibitors, as described, may include drugs that block or inhibit immune checkpoint receptors, and optionally, immune checkpoint receptors include adenosine A2A receptor (A2AR / ADORA2A), adenosine A2B receptor (A2BR / ADORA2B), B7 Homolog 3 (CD276 / B7-H3), V-set domain-containing T cell activation inhibitor 1 (VTCN1 / B7-H4), B and T lymphocyte attenuator (CD272 / BTLA), cytotoxic T lymphocyte-associated protein 4 (CD152 / CTLA-4), indoleamine-pyrrole 2,3-dioxygenase (IDO), killer cell immunoglobulin-like receptor (KIR), lymphocyte activation gene 3 (LAG-3), and NADPH oxidase. 2 (NOX2 / cytochrome b(558) subunit beta / cytochrome b-245 heavy chain), programmed cell death protein 1 (PD-1), T cell immunoglobulin and mucin domain-containing 3 (TIM-3 / hepatitis A virus cell receptor 2 [HAVCR2]), T cell activation V-domain Ig suppressor (VISTA), sialic acid-binding Ig-like lectin 7 (SIGLEC7), and optionally, one or more of their ligands may be selected.

[0038] In some embodiments, immune checkpoint inhibitors as described herein include programmed cell death protein 1 (PD-1) and cytotoxic T lymphocyte-associated protein 4 (CD152 / CTLA-4), and optionally, drugs that block or inhibit immune checkpoint receptors selected from their ligands.

[0039] In some embodiments, the immune checkpoint inhibitors described herein include drugs that block or inhibit immune checkpoint receptor programmed cell death protein 1 (PD-1). In some embodiments, the immune checkpoint inhibitors described herein include drugs that block or inhibit a ligand of immune checkpoint receptor programmed cell death protein 1 (PD-1) selected from programmed cell death protein ligand 1 (PD-1) and programmed cell death protein ligand 2 (PD-2).

[0040] In some embodiments, the immune checkpoint inhibitors described herein include monoclonal antibodies that target and bind to immune checkpoint receptors selected from programmed cell death protein 1 (PD-1) and cytotoxic T lymphocyte-associated protein 4 (CTLA-4) (Kim SK, Cho SW. The Evasion Mechanisms of Cancer Immunity and Drug Intervention in the Tumor Microenvironment. Front Pharmacol. 2022 May 24;13:868695. doi: 10.3389 / fphar.2022.868695. PMID: 35685630; PMCID: PMC9171538).

[0041] In some embodiments, the immune checkpoint inhibitor as described herein includes a monoclonal antibody that targets and binds to programmed cell death protein 1 (PD-1). In some embodiments, the immune checkpoint inhibitor as described herein includes a monoclonal antibody that targets and binds to a ligand of programmed cell death protein 1 (PD-1) selected from programmed cell death protein ligand 1 (PD-1) and programmed cell death protein ligand 2 (PD-2).

[0042] As used herein, the term "thinostamstine" refers to a compound having the following structural formula:

[0043] [ka]

[0044] It refers to.

[0045] "Pharmacologically acceptable salts" means salts of the compounds of the present invention (e.g., tinostamustine) that are pharmaceutically acceptable and retain the desired pharmacological activity. Such salts include acid addition salts formed with inorganic acids or organic acids. Pharmaceutically acceptable salts also include base addition salts that can be formed when the present acidic protons can react with inorganic or organic bases. Generally, such salts are prepared by reacting the free acid or base form of these compounds with a stoichiometric amount of a suitable base or acid, for example, in water, or in an organic solvent, or in a mixture of the two. Generally, non-aqueous media such as ether, ethyl acetate, ethanol, isopropanol, or acetonitrile are preferred. Examples of acid addition salts include inorganic acid addition salts such as hydrochloride, hydrobromide, hydroiodide, sulfate, bisulfate, sulfamate, nitrate, and phosphate, as well as organic acid addition salts such as acetate, trifluoroacetate, maleate, fumarate, citrate, oxalate, succinate, tartrate, salicylate, tosylate, lactate, naphthalene sulfonate, malate, mandelate, methanesulfonate, and p-toluenesulfonate. Examples of alkali addition salts include inorganic salts such as sodium, potassium, calcium, and ammonium salts, as well as organic alkali salts such as ethylenediamine, ethanolamine, N,N-dialkyleneethanolamine, triethanolamine, and basic amino acid salts.

[0046] As used herein, the term “antibody” refers to any immunoglobulin, preferably full-length immunoglobulin, unless otherwise specified. Preferably, the term extends to monoclonal antibodies, polyclonal antibodies, bispecific antibodies, and other multispecific antibodies, and antibody fragments thereof, insofar as they exhibit the desired biological activity. Antibodies may originate from any species, but are preferably mouse, human, or rabbit. Alternatively, antibodies, preferably monoclonal antibodies, may be humanized, chimeric, or antibody fragments thereof. The term “chimeric antibody” may also include “primatised” antibodies containing variable domain antigen-binding sequences and human constant region sequences derived from non-human primates (e.g., Old World monkeys, apes, etc.). Immunoglobulins may also be any type (e.g., IgG, IgE, IgM, IgD, and IgA), class (e.g., IgG1, IgG2, IgG3, IgG4, IgAl, and IgA2), or subclass of immunoglobulin molecules.

[0047] As used herein, the term “monoclonal antibody” refers, unless otherwise specified, to a substantially homogeneous population of antibody molecules produced by a single clone of B lineage cells, often a hybridoma, where the individual antibodies constituting the population are identical except for possible naturally occurring mutations that may be present in small amounts. Importantly, each monoclonal antibody has the same antigen specificity, i.e., it is directed toward a single determinant on the antigen.

[0048] As used herein, the term “to treat” means, unless otherwise indicated, to reverse, reduce, alleviate or inhibit the progression of a disease or condition, or one or more symptoms of such disorder or condition, to which such term applies. As used herein, the term “to treat” means, unless otherwise indicated, the act of treating as defined above.

[0049] As used herein, the term “patient” includes humans, non-human mammals (e.g., dogs, cats, rabbits, cattle, horses, sheep, goats, pigs, deer, and the like) and non-mammals (e.g., birds, and the like). Preferably, the patient is a human patient.

[0050] As used herein, the term “progressive” in relation to cancer (e.g., progressive melanoma) means, unless otherwise indicated, cancer that is not curable or controllable by treatment (i.e., clinically approved treatment). Cancer may spread from its original site (primary tumor) to other areas of the body and may therefore be metastatic cancer. Advanced cancer may be referred to as terminal cancer or palliative cancer.

[0051] As used herein, the term “recurrent” in relation to cancer means, unless otherwise specified, cancer that has recurred after a period of remission in response to a preceding line of treatment (either partial response [PR] or complete response [CR]), but has recurred after the termination of the preceding line of treatment. Recurrent cancer is sometimes referred to as “recurrent” cancer.

[0052] As used herein, the term “refractory” in relation to cancer means cancer that has not responded to one or more prior lines of treatment, unless otherwise specified. Refractory cancers are sometimes referred to as “resistant” to standard treatment.

[0053] As used herein, the term “adjuvant” in relation to cancer treatment means, unless otherwise specified, a treatment administered to a patient who requires it to reduce the risk of cancer recurrence, following the administration of primary treatment (e.g., chemotherapy, radiation therapy, surgery).

[0054] As used herein, the term “neoadjuvant” in relation to cancer treatment means, unless otherwise specified, a treatment administered to a patient who requires it to reduce tumor volume before the administration of the primary treatment (e.g., chemotherapy, radiotherapy, surgery). For example, if the primary treatment is chemotherapy (e.g., immunotherapy), the neoadjuvant may be administered to induce a biological response (e.g., an immunological response) that enhances the effectiveness of the primary treatment.

[0055] To provide a more concise explanation, some of the quantitative expressions presented herein are not modified with the term “approximately.” Whether the term “approximately” is explicitly used or not, any quantity presented herein is meant to refer to the actual value shown, and also to an approximation of the such value that could be reasonably inferred on the basis of ordinary skill in the art, including equivalents and approximations resulting from experimental and / or measurement conditions for the such value.

[0056] Detailed explanation The following detailed description relates to programmed cell death protein 1 (PD-1) and programmed cell death protein ligand 1 (PD-L1) as exemplary immune checkpoint inhibitors, but those skilled in the art will recognize that the scope of the invention is not limited to these particular classes of immune checkpoint inhibitors, and that the benefits of the invention can be understood by providing combinations including any suitable immune checkpoint inhibitors.

[0057] As described above, the interaction between PD-1 and its receptor PD-L1 (and / or PD-L2) is involved in promoting tumor cell survival and is therefore an attractive target for novel immunotherapies. PD-L1 (and PD-L2) has been shown to be upregulated in a variety of cancers, including solid tumors such as melanoma, lung cancer, colorectal cancer, gastric cancer, bladder cancer, pancreatic cancer, prostate cancer, ovarian cancer, breast cancer, and non-Hodgkin lymphoma (Han Y, Liu D, Li L. PD-1 / PD-L1 pathway: current researches in cancer. Am J Cancer Res. 2020 Mar 1;10(3):727~742. PMID: 32266087; PMCID: PMC7136921), and immune checkpoint inhibitors targeting this target have been widely investigated.

[0058] Combination therapies involving one or more immune checkpoint inhibitors have been shown to be more effective in treating cancer than immune checkpoint inhibitors as monotherapy. For example, combination immunotherapies involving immune checkpoint inhibitors targeting PD-1 and CTLA-4 (e.g., nivolumab, an anti-PD-1 antibody, combined with the anti-CTLA4 antibody ipilimumab) are clinically approved for treating cancer. For example, a recent clinical review (Johnson et al., Immune-checkpoint inhibitors: long-term implications of toxicity. Nat Rev Clin Oncol 19, pp. 254-267 (2022). https: / / doi.org / 10.1038 / s41571-022-00600-w) suggests that treatment with nivolumab in combination with ipilimumab is associated with a response rate of 59% in metastatic melanoma (compared to 43% with nivolumab alone and 15-20% with ipilimumab alone), and approximately 40% in renal cell carcinoma (RCC) (compared to approximately 25% with nivolumab alone and minimal activity with ipilimumab as monotherapy). This combination is also approved for use in patients with non-small cell lung cancer (NSCLC), highly microsatellite-instability colorectal cancer, or hepatocellular carcinoma in whom CTLA-4 inhibition is characterized as having little (or insufficient) activity as monotherapy.

[0059] However, parallel inhibition of immune checkpoints such as PD-1 and CTLA-4 simultaneously increases the risk of autoimmune toxicity. Such combination therapies have been observed to lead to an increased incidence of severe immune-related adverse events (irAEs). Specifically, nivolumab monotherapy, ipilimumab monotherapy, and the combination of ipilimumab and nivolumab have been shown to cause severe adverse events in 23%, 28%, and 59% of patients with advanced melanoma, respectively (Johnson et al., Immune-checkpoint inhibitors: long-term implications of toxicity. Nat Rev Clin Oncol 19, pp. 254-267 (2022). https: / / doi.org / 10.1038 / s41571-022-00600-w).

[0060] The present invention addresses this problem by providing a combination according to a first aspect of the present invention. Surprisingly, the combination disclosed herein not only avoids adverse events in patients associated with existing combinations including immune checkpoint inhibitors, but also improves overall survival and progression-free survival in the clinical patient population, as will be described in more detail in the examples below. Therefore, the present invention is a promising treatment for use in the treatment of cancer, reducing or even eliminating adverse events in patients receiving current standard treatments including combinations of immune checkpoint inhibitors, and further improving clinical outcomes.

[0061] Furthermore, the combinations described herein according to a first aspect of the present invention have been shown to be effective in increasing overall survival and progression-free survival in patients with progressive diseases, i.e., patients who have received one or more prior lines of treatment and whose diseases are not considered likely to respond to continued treatment using lines of current standard care. Surprisingly, the examples herein illustrate observations of increased progression-free survival and overall survival on a scale of several months, representing a significant delay in disease progression onset and death.

[0062] In some embodiments, pharmaceutically acceptable salts of tinostamstine by compositions and methods provided herein are hydrochloride, hydrobromide, methanesulfonate, toluenesulfonate, acetate, fumarate, sulfate, bisulfate, succinate, citrate, phosphate, maleate, nitrate, tartrate, benzoate, biocarbonate, carbonate, sodium hydroxide, calcium hydroxide, potassium hydroxide, tromethamine, or mixtures thereof.

[0063] In some embodiments, an immune checkpoint inhibitor blocks or substantially blocks the binding of PD-L1 to PD-1, for example, by binding to PD-1. In one embodiment, an immune checkpoint inhibitor is any agent or drug that can bind to PD-1 and block or substantially block the binding of PD-L1 or PD-1. Examples of such agents include peptides, cyclic peptides, small molecules / pharmaceuticals, antibodies, functional fragments of antibodies, nanobodies, or aptamers.

[0064] According to a first aspect of the present invention, the immune checkpoint inhibitor may be an anti-PD-1 antibody optionally selected from pembrolizumab, nivolumab, dostallimab, retifanlimab, and semiprimab.

[0065] In some embodiments, the immune checkpoint inhibitor is pembrolizumab. In such embodiments, the claimed combination according to the present invention may be used in the treatment of cancer selected from melanoma, non-small cell lung cancer, Hodgkin lymphoma, urothelial carcinoma, head and neck squamous cell carcinoma, and renal cell carcinoma.

[0066] In some embodiments, the immune checkpoint inhibitor is nivolumab. In such embodiments, the claimed combination according to the present invention may be used in the treatment of cancers selected from melanoma, non-small cell lung cancer, malignant pleural mesothelioma, advanced renal cell carcinoma, Hodgkin lymphoma, squamous cell carcinoma of the head and neck, urothelial carcinoma, metastatic colorectal cancer with high microsatellite instability or mismatch repair deficiency, hepatocellular carcinoma, esophageal cancer, gastric cancer, gastroesophageal junction cancer, and esophageal adenocarcinoma.

[0067] In some embodiments, the immune checkpoint inhibitor is dostallimab. In such embodiments, the claimed combination of the present invention may be used in the treatment of cancers selected from endometrial cancer, colorectal cancer and solid tumors (e.g., sarcomas and carcinomas), optionally from advanced solid tumors.

[0068] In some embodiments, the immune checkpoint inhibitor is retifanlimab. In such embodiments, the claimed combination according to the present invention may be used in the treatment of cancer selected from advanced Merkel cell carcinoma.

[0069] In some embodiments, the immune checkpoint inhibitor is cemiplimab. In such embodiments, the claimed combination according to the present invention may be used in the treatment of cancers selected from melanoma, metastatic cutaneous squamous cell carcinoma (CSCC), myeloma, lung cancer, and cervical cancer.

[0070] In some embodiments, the immune checkpoint inhibitor is a programmed cell death protein ligand 1 (PD-L1) inhibitor and / or a programmed cell death protein ligand 2 (PD-L2) inhibitor. In some embodiments, the immune checkpoint inhibitor is a PD-L1 inhibitor.

[0071] In some embodiments, the immune checkpoint inhibitor is an anti-PD-L1 antibody selected at least selectively from atezolizumab, avelumab, and durvalumab.

[0072] In some embodiments, the immune checkpoint inhibitor is atezolizumab. In such embodiments, the claimed combination according to the present invention may be used in the treatment of cancer selected from melanoma, urothelial carcinoma, small cell lung cancer, non-small cell lung cancer, hepatocellular carcinoma, bladder cancer, renal cell carcinoma, and trinegative breast cancer.

[0073] In some embodiments, the immune checkpoint inhibitor is avelumab. In such embodiments, the claimed combination according to the present invention may be used in the treatment of cancer selected from melanoma, urothelial carcinoma, Merkel cell carcinoma, and renal cell carcinoma.

[0074] In some embodiments, the immune checkpoint inhibitor is durvalumab. In such embodiments, the claimed combination according to the present invention may be used in the treatment of cancer selected from melanoma, urothelial carcinoma, non-small cell lung cancer, small cell lung cancer, and cholangiocarcinoma.

[0075] In some embodiments, the combination may further include one or more additional immune checkpoint inhibitors.

[0076] For example, the combination may further include a cytotoxic T lymphocyte-associated antigen 4 (CTLA-4) inhibitor, which may optionally be selected from ipilimumab and tremelimumab, and optionally be ipilimumab.

[0077] In some embodiments, the combination further comprises an additional immune checkpoint inhibitor, the additional immune checkpoint inhibitor being ipilimumab. In such embodiments, the claimed combination according to the present invention may be used in the treatment of cancers selected from melanoma, renal cell carcinoma, colorectal cancer, hepatocellular carcinoma, non-small cell lung cancer, malignant pleural mesothelioma, prostate cancer, and urothelial carcinoma.

[0078] In some embodiments, the combination further comprises an additional immune checkpoint inhibitor, the additional immune checkpoint inhibitor being tremelimumab. In such embodiments, the claimed combination according to the present invention may be used in the treatment of cancers selected from melanoma (e.g., advanced melanoma), mesothelioma, and non-small cell lung cancer.

[0079] The combination according to the first aspect of the present invention may further include one or more additional pharmaceutically active agents. Particularly preferred pharmaceutically active agents are antitumor agents having a different mechanism of action than tinostamstine and / or immune checkpoint inhibitors. Preferred antitumor agents include alkylating agents, e.g., nitrosurea, ethyleneimine, alkyl sulfonates, hydrazine and triazine, and platinum-based agents; plant alkaloids, taxanes, vinca alkaloids; antitumor antibiotics, e.g., chromomycin, anthracyclines, and various antibiotics, e.g., mitomycin and bleomycin; antimetabolites, e.g., folate antagonists, pyrimidine antagonists, purine antagonists, and Adenosine deaminase inhibitors; topoisomerase inhibitors, e.g., topoisomerase I inhibitors, topoisomerase II inhibitors; various antineoplasms, e.g., ribonucleotide reductase inhibitors, corticosteroid inhibitors, antimicrotubule agents, and retinoids; protein kinases; heat shock proteins, poly-ADP (adenosine diphosphate)-ribose polymerase (PARP), hypoxia-inducible factor (HIF), proteasomes, Wnt / Hedgehog / Notch signaling proteins, TNF-alpha, matrix metalloproteinases, farnesyltransferases, apoptotic pathways, histone deacetylases (HDAC), histone acetyltransferases (HAT), and methyltransferases; hormone therapy, vasolytic agents, gene therapy, RNAi cancer therapy, chemoprotective agents, antibody conjugates, cancer immunotherapy, e.g., interleukin-2, cancer vaccines, or monoclonal antibodies; and preferably DNA damage This may include drugs, antimetabolites, topoisomerase inhibitors, antimicrotubule agents, EGFR inhibitors, HER2 inhibitors, VEGFR2 inhibitors, BRAF inhibitors, Bcr-Abl inhibitors, PDGFR inhibitors, ALK inhibitors, PLK inhibitors, MET inhibitors, epigenetic drugs, HSP90 inhibitors, PARP inhibitors, CHK inhibitors, aromatase inhibitors, estrogen receptor antagonists, and antibodies targeting VEGF, HER2, EGFR, CD50, CD20, CD30, and CD33.

[0080] In one preferred embodiment of the combination of the present invention, the immune checkpoint inhibitor and tinostamustine are adapted for simultaneous, sequential, or separate administration. Preferably, the immune checkpoint inhibitor and tinostamustine are adapted for separate administration.

[0081] In some embodiments, tinostamustine is administered to the patient without an immune checkpoint inhibitor during the first cycle of treatment, and tinostamustine and an immune checkpoint inhibitor are administered to the patient sequentially, simultaneously, or separately during all subsequent cycles of treatment (e.g., second, third, etc.). For example, in some embodiments, tinostamustine is administered to the patient without an immune checkpoint inhibitor during the first cycle of treatment, and then, during the second cycle of treatment, tinostamustine and an immune checkpoint inhibitor are administered to the patient sequentially (i.e., one after the other without delay) or separately (i.e., the other after a certain period following the administration of one), and all subsequent cycles may follow the second cycle.

[0082] In embodiments where tinostamustine and an immune checkpoint inhibitor are administered separately, the immune checkpoint inhibitor may be administered to the patient first, followed by tinostamustine or a pharmaceutically acceptable salt thereof. In some embodiments, the time between the separate administration of the immune checkpoint inhibitor and tinostamustine or a pharmaceutically acceptable salt thereof is 20 to 120 minutes, optionally 30 to 90 minutes, preferably 50 to 70 minutes (e.g., 60 minutes). Preferably, the time between the separate administration of the immune checkpoint inhibitor and tinostamustine or a pharmaceutically acceptable salt thereof is at least 30 minutes.

[0083] In some embodiments, the patient is administered the treatment on the first day of each cycle. In some embodiments, the treatment cycle is at least 7 days, preferably at least 14 days (for example, the cycle is 14 days long). In some embodiments, the period between the first treatment cycle and the second treatment cycle is at least 7 days, preferably at least 14 days (for example, 14 days).

[0084] In some preferred embodiments, each cycle of treatment is 14 days long, and on day 1 of cycle 1, tinostamustine is administered to patients who require it, without an immune checkpoint inhibitor. In the second cycle of treatment, on day 1, tinostamustine and an immune checkpoint inhibitor are administered separately to patients who require them, with the immune checkpoint inhibitor administered first, followed by tinostamustine or a pharmaceutically acceptable salt thereof at least 30 minutes after the administration of the immune checkpoint inhibitor. In all subsequent cycles, on day 1 of each cycle, tinostamustine and an immune checkpoint inhibitor are administered separately to patients who require them, with the immune checkpoint inhibitor administered first, followed by tinostamustine or a pharmaceutically acceptable salt thereof at least 30 minutes after the administration of the immune checkpoint inhibitor. The immune checkpoint inhibitor may be a PD-1 inhibitor (e.g., nivolumab).

[0085] In some embodiments of the combination of the present invention, the molar ratio of immune checkpoint inhibitor to tinostamustine is 1:8000 to 1:500, optionally 1:7000 to 1:1000, preferably 1:6000 to 1:2000, and preferably even more preferably 1:5000 to 1:3000, for example, about 1:4000.

[0086] In some embodiments, combinations of an immune checkpoint inhibitor according to the present invention and tinostamustine, or a pharmaceutically acceptable salt thereof, may be synergistic combinations. For example, combinations of a PD-1 inhibitor and tinostamustine, or a pharmaceutically acceptable salt thereof, may be synergistic combinations.

[0087] According to a second aspect, the present invention further provides a pharmaceutical composition comprising a pharmaceutically acceptable diluent or carrier and a combination according to a first aspect of the present invention. Preferred compositions include preferred combinations of the present invention as described herein. The pharmaceutically acceptable diluent or carrier of a pharmaceutical composition according to a second aspect of the present invention may be any suitable dispersant, additive, adjuvant, or other substance that acts as a carrier for the active agent of the combination of the present invention and does not interfere with the active agent present in the combination. Typical examples of pharmaceutically acceptable carriers and diluents can be found in "Remington's Pharmaceutical Sciences" by E.W. Martin, and these include water, saline, dextrose solution, serum solution, Ringer's solution, polyethylene glycol (e.g., PEG400), surfactants (e.g., cremofor), cyclopolysaccharides (e.g., hydroxypropyl-β-cyclodextrin or sulfobutyl ether β-cyclodextrin), polymers, liposomes, micelles, nanoparticles, etc.

[0088] In some embodiments, the pharmaceutical composition containing tinostamstine also contains cyclodextrin. Cyclodextrin is a cyclic oligomer of dextrose having a shortened conical structure consisting of hydrophilic external pores and hydrophobic internal pores. Cyclodextrin can form inclusion complexes with guest molecules by complexing with all or some of the hydrophobic guest molecules within its pores. The size of the pores is determined by the number of glucopyranose units in the cyclodextrin. Alpha-(α), beta-(β), and gamma-(γ) cyclodextrins are the most common cyclodextrins, having 6, 7, and 8 glucopyranose units, respectively. Natural cyclodextrins have relatively low water solubility and are associated with toxicity; therefore, chemically modified cyclodextrin derivatives have been developed to overcome these limitations. Such cyclodextrin derivatives typically have chemical modifications at one or more of the 2, 3, or 6-position hydroxyl groups. Cyclodextrin derivatives are described, for example, in U.S. Patents 5,134,127; 5,376,645; 5,571,534; 5,874,418; 6,046,177 and 6,133,248. As used herein, the terms “cyclodextrin,” “α-cyclodextrin,” “β-cyclodextrin,” and “γ-cyclodextrin” are intended to encompass unmodified cyclodextrins and their chemically modified derivatives.

[0089] In some embodiments, the composition comprises a cyclodextrin selected from the group consisting of α-cyclodextrin, β-cyclodextrin, and γ-cyclodextrin. In a particular embodiment, the cyclodextrin is β-cyclodextrin. In a further embodiment, the cyclodextrin is selected from the group consisting of hydroxypropyl-β-cyclodextrin (Pitha et al., J Pharm Sci, 84 (8), pp. 927-932 (1995)) and sulfobutyl-derivative-β-cyclodextrin (e.g., as described in U.S. Patents 5,134,127; 5,376,645; 5,874,418; 6,046,177 and 6,133,248). In another embodiment, the cyclodextrin is hydroxypropyl-β-cyclodextrin. In yet another embodiment, the cyclodextrin is sulfobutyl ether-β-cyclodextrin. Other preferred cyclopolysaccharides include, but are not limited to, β-cyclodextrins substituted with 2-hydroxy-N,N,N-trimethylpropaneammonium, carboxymethylated β-cyclodextrins, O-phosphorylated β-cyclodextrins, succinyl-(2-hydroxyl)propyl-beta-cyclodextrins, sulfopropylated β-cyclodextrins, heptakis(6-amino-6-deoxy)-β-cyclodextrins, O-sulfated β-cyclodextrins, and 6-monodeoxy-6-mono(3-hydroxy)propylamino-β-cyclodextrins.

[0090] In some embodiments, cyclodextrin is included in an amount that increases the solubility of the active compound in the composition. In one embodiment, the amount of cyclodextrin included in the composition is the minimum amount required to dissolve the drug in the composition. In a further embodiment, the composition is a parenteral formulation, and the amount of cyclodextrin included in the formulation is the minimum amount of cyclodextrin required to dissolve the drug.

[0091] In some embodiments, the composition contains at least 2.5%, at least 5%, or at least 10% (by weight / volume) of cyclodextrin. In some other embodiments, the composition contains 2.5–40%, 5–20%, or 7.5–15% of cyclodextrin. In yet another embodiment, the composition contains about 10% of cyclodextrin.

[0092] In some embodiments, the composition further comprises a pH adjuster. In some further embodiments, the pH adjuster is one or more acids, bases, or salts. Examples of acids that may be included in the composition include inorganic acids such as hydrochloric acid, sulfuric acid, phosphoric acid, or mixtures thereof, and organic acids such as citric acid, L(-)-malic acid and L(+) tartaric acid, or mixtures thereof. Examples of bases that may be included in the composition include sodium hydroxide, potassium hydroxide, calcium hydroxide, tromethamine, or mixtures thereof. Examples of salts that may be included in the composition include sodium bicarbonate, sodium carbonate, sodium citrate, or mixtures thereof. In one further embodiment, the composition comprising one or more pH adjusters has a pH range of 6.0 to 9.0, preferably 7.0 to 8.0.

[0093] In some further embodiments, the composition contains dextran. In another embodiment, the composition contains dextran in an amount ranging from about 1% to about 5% by weight / volume. In yet another embodiment, the composition contains about 2% to about 4% by weight / volume of dextran.

[0094] Any inert additives commonly used as carriers or diluents, such as sugars, polyalcohols, soluble polymers, salts, and lipids, may be used in the compositions of the present invention. Examples of sugars and polyalcohols that may be used include, but are not limited to, lactose, sucrose, mannitol, and sorbitol. Examples of soluble polymers that may be used include polyoxyethylene, poloxamer, polyvinylpyrrolidone, and dextran. Useful salts include, but are not limited to, sodium chloride, magnesium chloride, and calcium chloride. Lipids that may be used include, but are not limited to, fatty acids, glycerol fatty acid esters, glycolipids, and phospholipids.

[0095] In addition, the composition further contains binders (e.g., gum arabic, corn starch, gelatin, carbomer, ethylcellulose, gum acacia, hydroxypropylcellulose, hydroxypropylmethylcellulose, povidone), disintegrants (e.g., corn starch, potato starch, alginic acid, silicon dioxide, croscarmellose sodium, crospovidone, gum acacia, sodium starch glycolate, Primogel), buffers of various pH and ionic strengths (e.g., Tris-HCl, acetate, phosphate), additives such as albumin or gelatin to prevent absorption to the surface, and detergents (e.g., Tween 20, Tween 80, Pluronic). F68 (bile salt), protease inhibitors, surfactants (e.g., sodium lauryl sulfate), penetration enhancers, solubilizers (e.g., glycerol, polyethyleneglycerol, cyclodextrin), fluidity enhancers (e.g., colloidal silicon dioxide), antioxidants (e.g., ascorbic acid, sodium metabisulfite, butylated hydroxyanisole), stabilizers (e.g., hydroxypropyl cellulose, hydroxypropyl methylcellulose), thickeners (e.g., carbomer, colloidal silicon dioxide, ethylcellulose, gargomuth), sweeteners (e.g., sucrose, aspartame, citric acid), flavorings (e.g., peppermint, salicylic acid) It may contain methyl cylate or orange flavoring agents, preservatives (e.g., thimerosal, benzyl alcohol, parabens), lubricants (e.g., stearic acid, magnesium stearate, polyethylene glycol, sodium lauryl sulfate), fluidizing agents (e.g., colloidal silicon dioxide), plasticizers (e.g., diethyl phthalate, triethyl citrate), emulsifiers (e.g., carbomer, hydroxypropyl cellulose, sodium lauryl sulfate), polymer coatings (e.g., poloxamer or poloxamine), coating and film-forming agents (e.g., ethyl cellulose, acrylate, polymethacrylate), and / or adjuvants.

[0096] In some embodiments, the composition is prepared with a carrier that will protect the compound from rapid elimination from the body, such as a controlled-release formulation including implants and microencapsulation delivery systems. Biodegradable, biocompatible polymers such as ethylene vinyl acetate, polyanhydride, polyglycolic acid, collagen, polyorthoesters, and polylactic acid may be used. Methods for preparing such formulations will be obvious to those skilled in the art. These materials are also commercially available from Alza Corporation and Nova Pharmaceuticals, Inc. Liposome suspensions (containing liposomes targeting infected cells, including monoclonal antibodies against viral antigens) may also be used as pharmaceutically acceptable carriers. These can be prepared by methods known to those skilled in the art, for example, as described in U.S. Patent No. 4,522,811.

[0097] According to a third aspect, the present invention further provides a kit comprising a combination according to the first aspect of the present invention and, optionally, instructions for treating a patient. Typically, the kit may include tinostamustine, or a pharmaceutically acceptable salt thereof, and an immune checkpoint inhibitor, together with instructions for treating a patient. Each active agent may be provided in a suitable container. The kit may further include, for example, a delivery system for tinostamustine, or a pharmaceutically acceptable salt thereof, or an immune checkpoint inhibitor, or any combination thereof.

[0098] The instructions may advise on the combination of tinostammustine or pharmaceutically acceptable salts thereof, or immune checkpoint inhibitors, simultaneously, sequentially, or separately, according to variables such as the specific condition being treated, the circumstances of that condition, the activity of the specific compound used; the specific combination used; the patient's age, weight, general health, sex, and diet; the timing of administration, route of administration, and excretion rate of the specific compound used; the duration of treatment; drugs used in combination with or concurrently with the specific compound used; and similar factors well known in the medical field. A preferred kit according to a third aspect of the present invention includes one containing the preferred combination of the present invention as described herein.

[0099] According to a fourth aspect of the present invention, a combination, composition, or kit according to the first, second, or third aspect of the present invention is provided for use in the treatment of cancer.

[0100] A fifth aspect of the present invention provides tinostamustine, or a pharmaceutically acceptable salt thereof, for use in the treatment of cancer, in combination with an immune checkpoint inhibitor.

[0101] According to a sixth aspect of the present invention, an immune checkpoint inhibitor is provided for use in the treatment of cancer, wherein the immune checkpoint inhibitor is used in combination with tinostamstine or a pharmaceutically acceptable salt thereof.

[0102] A seventh aspect of the present invention provides a method for treating cancer in a patient in need, comprising administering to the patient a combination, composition, or kit according to the first, second, or third aspect of the present invention. The method comprises administering to the patient in need a therapeutically effective dose of tinostamustine, or a pharmaceutically acceptable salt thereof, in combination with a therapeutically effective dose of an immune checkpoint inhibitor.

[0103] The combinations, compositions, and kits of the present invention have been found to be effective in the treatment of cancer, particularly advanced cancer, and more particularly advanced melanoma.

[0104] The combinations, compositions, and kits of the present invention have also been found to be effective in the treatment of colorectal cancer, particularly colorectal carcinoma.

[0105] In some preferred embodiments, the cancer is recurrent and / or refractory. In some preferred embodiments, the cancer is metastatic. In some preferred embodiments, the cancer is unresectable.

[0106] Cancers treatable with the combinations, compositions, and kits of the present invention include hematological cancers (such as multiple myeloma, lymphoma, and leukemia), breast cancer, lung cancer, colorectal cancer, prostate cancer, testicular cancer, pancreatic cancer, liver cancer, gastric cancer, bile duct cancer, esophageal cancer, gastrointestinal stromal tumors, cervical cancer, ovarian cancer, uterine cancer, kidney cancer, melanoma, basal cell carcinoma, squamous cell carcinoma, bladder cancer, sarcoma, mesothelioma, thymoma, myelodysplastic syndromes, brain cancers (e.g., glioblastoma), and myeloproliferative disorders.

[0107] Cancers may be selected from melanoma, small cell lung cancer, non-small cell lung cancer, mesothelioma (e.g., malignant pleural mesothelioma), renal cell carcinoma, head and neck cancer, urothelial carcinoma, colorectal cancer, esophageal squamous cell carcinoma, hepatocellular carcinoma, gastric cancer, esophageal cancer, esophageal adenocarcinoma, gastroesophageal junction cancer, high-frequency microsatellite instability or mismatch repair deficiency cancer, primary mediastinal large B-cell lymphoma (PMBCL), Merkel cell carcinoma (MCC), endometrial cancer, high tumor mutational load (TMB-H) cancer, cutaneous squamous cell carcinoma (cSCC), soft tissue sarcoma, osteosarcoma, ovarian cancer, trinegative breast cancer, brain cancer (e.g., glioblastoma), multiple myeloma, Hodgkin lymphoma, and non-Hodgkin lymphoma.

[0108] Cancers may be selected from melanoma, small cell lung cancer, non-small cell lung cancer, mesothelioma (e.g., malignant pleural mesothelioma), renal cell carcinoma, head and neck cancer, urothelial carcinoma, colorectal cancer (e.g., MSI-H or dMMR metastatic colorectal cancer), esophageal squamous cell carcinoma, hepatocellular carcinoma, gastric cancer, esophageal cancer, esophageal adenocarcinoma, gastroesophageal junction cancer, high microsatellite instability or mismatch repair deficiency cancer, primary mediastinal large B-cell lymphoma (PMBCL), Merkel cell carcinoma (MCC), endometrial cancer, high tumor mutational load (TMB-H) cancer, and cutaneous squamous cell carcinoma (cSCC).

[0109] The cancer may be selected from melanoma, soft tissue sarcoma, osteosarcoma, ovarian cancer, trinegative breast cancer, brain cancer (e.g., glioblastoma), multiple myeloma, Hodgkin lymphoma, and non-Hodgkin lymphoma.

[0110] Cancers may be selected from cancers for which immune checkpoint inhibitors are clinically approved (e.g., by the U.S. Food and Drug Administration; or the European Medicines Agency) for use in the treatment of said cancers. For example, cancers may be selected from cancers for which PD-1 or PD-L1 inhibitors are clinically approved for use in the treatment of said cancers. For example, cancers may be selected from cancers for which nivolumab is clinically approved for use in the treatment of said cancers (e.g., melanoma, non-small cell lung cancer, malignant pleural mesothelioma, renal cell carcinoma, Hodgkin lymphoma, squamous cell carcinoma of the head and neck, urothelial carcinoma, MSI-H or dMMR metastatic colorectal cancer, hepatocellular carcinoma, esophageal cancer, gastric cancer, and gastroesophageal junction cancer).

[0111] When the combination, composition, or kit of the present invention is intended for use in the treatment of hematological cancers, it is preferably multiple myeloma (e.g., active myeloma, plasma cell tumor, light chain myeloma, or non-secretory myeloma, including all treatable forms in all phases, including relapsed and refractory phases), lymphoma (e.g., Hodgkin lymphoma or non-Hodgkin lymphoma) and leukemia [acute lymphoblastic leukemia (ALL), chronic lymphocytic leukemia (CLL)]. The following may be selected from acute myeloid leukemia (including all treatable forms in all phases, including relapsed and refractory phases, including myeloblastic leukemia, acute promyelocytic leukemia, acute myeloid monocytic leukemia, acute monocytic leukemia, acute erythroleukemia and acute megakaryoblastic leukemia, AML), chronic myeloid leukemia (CML), hairy cell leukemia (HCL), T-cell prelymphoblastic leukemia (T-PLL), macrogranular lymphocytic leukemia, or T-cell acute lymphoblastic leukemia.

[0112] If the combination, composition, or kit of the present invention is intended for use in the treatment of brain cancer, optionally, the brain cancer is a glioma or glioblastoma, and optionally, the glioblastoma may be preferably selected from MGMT-positive astrocytic brain tumor, MGMT-negative astrocytic brain tumor, metastatic brain cancer, and primary CNS lymphoma. The glioblastoma may be an MGMT-positive astrocytic brain tumor. The glioblastoma may be an MGMT-negative astrocytic brain tumor.

[0113] In particular, the combinations, compositions and kits of the present invention are effective against melanoma, particularly advanced melanoma. When the combination, composition or kit of the present invention is for use in the treatment of melanoma, the melanoma may preferably be selected from acral lentiginous type, mucosal type, uveal type, nodular type, and superficial spreading melanoma.

[0114] In particular, the combinations, compositions or kits of the present invention are effective against colorectal cancer. When the combination, composition or kit of the present invention is for use in the treatment of colorectal cancer, the colorectal cancer may preferably be selected from carcinoma, primary colorectal lymphoma, gastrointestinal stromal tumor, leiomyosarcoma, carcinoid tumor and melanoma, and even more preferably, the colorectal cancer is carcinoma, and preferably the carcinoma is adenocarcinoma.

[0115] According to the 4th, 5th, 6th and 7th aspects of the present invention, chinostamustine or a pharmaceutically acceptable salt thereof is typically administered to a patient who needs it in a dosage range of 10 - 50 mg / patient body surface area m 2 , optionally in a dosage range of 20 - 40 mg / patient body surface area m 2 , preferably in a dosage range of 25 - 40 mg / patient body surface area m 2 , more preferably in a dosage range of 25 - 35 mg / patient body surface area m 2 (e.g., 30 mg / m 2 ). Chinostamustine or a pharmaceutically acceptable salt thereof may be administered to a patient who needs it in a dosage range of 10 - 50 mg / patient body weight kg, optionally in a dosage range of 20 - 40 mg / patient body weight kg, preferably in a dosage range of 25 - 40 mg / patient body weight kg, more preferably in a dosage range of 25 - 35 mg / patient body weight kg.

[0116] Advantageously, it has been found that the administration of chinostamustine to a patient who needs it within the dosage ranges shown above does not have a harmful effect on the immune system and, in particular, does not cause significant myelosuppression. In some preferred embodiments, the dosage range of chinostamustine is 40 mg / patient body surface area m 2 since myelosuppression is not caused when it is less than, and the dosage range of chinostamustine is 40 mg / patient body surface area m 2The doses shall not exceed [amount]. Therefore, the use of tinostamustine, or a pharmaceutically acceptable salt thereof, within the indicated dosage range shall not adversely affect the intended mechanism of action of the immune checkpoint inhibitors included in the claimed combination. More specifically, the dosage range of tinostamustine described herein is preferable in which tinostamustine may avoid causing cytotoxicity to immune cells. For comparison, the doses of tinostamustine administered according to the present invention are less than the doses typically used to achieve anti-cancer neoplasm activity (e.g., by inducing cytotoxicity in tumor cells) with tinostamustine as monotherapy (typically 80–120 mg / kg).

[0117] According to the fourth, fifth, sixth, and seventh aspects of the present invention, an immune checkpoint inhibitor may typically be administered to a patient in need in a dosage range of 1 to 5 mg / kg of patient body weight, preferably 2 to 4 mg / kg of patient body weight, and more preferably about 3 mg / kg of patient body weight (e.g., 3 mg / kg). The immune checkpoint inhibitor may be administered to a patient in need at a dosage range of 1 to 5 mg / m² of patient body surface area. 2 Within this dosage range, preferably 2-4 mg / patient body surface area m² 2 Preferably, an additional approximately 3 mg / patient body surface area m² 2 (For example, 3 mg / m²) 2 It may be administered within the dosage range of ).

[0118] The therapeutically effective dose of a combination, composition, or kit according to the present invention is the amount of the combination, composition, or kit that imparts the therapeutic effect according to the fourth, fifth, sixth, and seventh aspects of the present invention to the subject being treated in a reasonable benefit / risk ratio applicable to any medical treatment. The therapeutic effect may be objective (i.e., measurable by several tests or markers) or subjective (i.e., the subject shows signs of or feels the effect). The effective dose of a combination, composition, or kit according to the present invention is a dosage range of 10-50 mg / kg of patient body weight (e.g., 20-40 mg / m² of body surface area) in the combination of tinostamustine or a pharmaceutically acceptable salt thereof. 2For example, 25, 30, or 35 mg / m² of body surface area. 2 It is thought that this includes immune checkpoint inhibitors in a dosage range of 1-5 mg / kg of patient body weight (e.g., 2-4 mg / kg of body weight, e.g., 3 mg / kg of body weight).

[0119] The effective dose will vary depending on the route of administration and the possibility of concomitant use with other active agents. However, it will be understood that the total daily dose of the combinations, compositions, and kits of the present invention will be determined by the attending physician within the bounds of appropriate medical judgment. The specific therapeutic effective dose level for any particular patient will depend on a variety of factors, including the disorder being treated (e.g., cancer) and its severity; the activity of the specific compound used; the specific composition used; the patient's age, weight, overall health, sex, and diet; the timing, route of administration, and excretion rate of the specific compound used; the duration of treatment; drugs used in combination with or concurrently with the specific compound used; and similar factors well known in the medical field.

[0120] In some embodiments, the patient has already received at least one line of treatment, meaning the patient has already undergone a course of treatment (e.g., chemotherapy, radiation therapy, surgery), but the cancer is still present in the patient. In some embodiments, the patient has a progressive disease and the tumor burden is increasing (e.g., a larger tumor volume and / or the cancer is spreading to other areas of the body).

[0121] In some embodiments, the patient has received at least one prior line of treatment. In some embodiments, the patient has received at least two prior lines of treatment. In some embodiments, the patient has received at least three prior lines of treatment. In some embodiments, the patient has received at least four prior lines of treatment. In some embodiments, the patient has received at least five prior lines of treatment.

[0122] In some embodiments, the patient has been treated with at least one prior line of treatment, which includes the administration of an immune checkpoint inhibitor. In such embodiments, preferably, the immune checkpoint inhibitor has not been administered to the patient for at least six months prior to initiating treatment according to the claimed invention. In some embodiments, the patient has not previously received an immune checkpoint inhibitor in a prior line of treatment.

[0123] Preferred forms of administration of the combinations, compositions, or kits of the present invention include, but are not limited to, oral, topical, parenteral, sublingual, rectal, vaginal, ocular, and intranasal administration. Parenteral administration includes subcutaneous injection, intravenous, intramuscular, intrasternal injection, or infusion techniques. Preferably, the combinations, compositions, or kits are administered parenterally. The combinations and compositions of the present invention may be formulated to enable the combinations or compositions of the present invention to be bioavailable for administration to animals, preferably humans. The compositions may take the form of one or more dosing units; for example, a tablet may be a single dosing unit, and a container of the combinations or compositions of the present invention in aerosol form may hold multiple dosing units.

[0124] Preferably, the combinations of the present invention are provided in the form of a kit. Typically, the kit comprises an immune checkpoint inhibitor and tinostamustine or a pharmaceutically acceptable salt thereof. In certain embodiments, the kit may comprise one or more delivery systems, e.g., an immune checkpoint inhibitor, tinostamustine or a pharmaceutically acceptable salt thereof, and instructions for using the kit (e.g., instructions for treating a subject). These instructions / instructions may advise administering the combination of the immune checkpoint inhibitor and tinostamustine or a pharmaceutically acceptable salt thereof simultaneously, sequentially, or separately, according to variables such as the specific condition being treated, the circumstances of that condition, the activity of the specific compounds used; the specific combination used; the patient's age, weight, overall health, sex, and diet; the timing of administration, route of administration, and excretion rate of the specific compounds used; the duration of treatment; drugs used in combination with or concurrently with the specific compounds used; and similar factors well known in the medical field.

[0125] Since pharmaceutically acceptable diluents or carriers may be fine particles, the compositions may be, for example, in tablet or powder form. The carrier may be a liquid, and the combination, composition, or kit may be, for example, an oral syrup or an injectable solution. In addition, the carrier may be a gas, which may provide an aerosol composition useful for, for example, in inhalation administration. Such pharmaceutical carriers may be liquids such as water and oil, including petroleum, animal, plant, or synthetic sources such as peanut oil, soybean oil, mineral oil, sesame oil, and the like. The carrier may be saline solution, gum arabic, gelatin, starch paste, talc, keratin, colloidal silica, urea, and the like. In addition, auxiliaries, stabilizers, thickeners, lubricants, and colorants may be used. In one embodiment, when administered to animals, the combination, composition, or kit of the present invention and the pharmaceutically acceptable carrier are sterile. When the combination or composition of the present invention is administered intravenously, water is a preferred carrier. Saline solutions and aqueous solutions of dextrose and glycerol may also be used as liquid carriers, particularly for injectable solutions. Suitable pharmaceutical carriers also include additives such as starch, glucose, lactose, sucrose, gelatin, malt, rice, wheat, chalk, silica gel, sodium stearate, glyceryl monostearate, talc, sodium chloride, skim milk powder, glycerol, propylene glycol, water, ethanol, and similar substances. The composition may also contain small amounts of wetting agents or emulsifiers, or pH buffers, if desired.

[0126] When intended for oral administration, the combination, composition, or kit may be in solid or liquid form, and semi-solid, semi-liquid, suspension, and gel forms are included within the range of forms that are determined to be either solid or liquid as herein.

[0127] As solid compositions for oral administration, the combinations, compositions, or kits may be formulated in the form of powders, granules, compressed tablets, pills, capsules, chewing gums, wafers, or similar forms. Such solid compositions typically contain one or more inert diluents as a single tablet containing all the active agents, or as several separate solid compositions (in the case of a kit) each containing one of the single active agents of the combination of the present invention. In addition, one or more of the following may be present: binders such as carboxymethylcellulose, ethylcellulose, microcrystalline cellulose, or gelatin; additives such as starch, lactose, or dextrin; disintegrants such as alginic acid, sodium alginate, corn starch, and the like; lubricants such as magnesium stearate; flow enhancers such as colloidal silicon dioxide; sweeteners such as sucrose or saccharin; flavorings such as peppermint, methyl salicylate, or orange flavorings; and colorings.

[0128] If the combination or composition is in the form of a capsule (e.g., a gelatin capsule), it may contain, in addition to the above types of substances, a liquid carrier such as polyethylene glycol, cyclodextrin, or fatty oil.

[0129] The combination, composition, or kit may be in the form of a liquid, such as an elixir, syrup, solution, emulsion, or suspension. The liquid may be useful for oral administration or for delivery by injection. When intended for oral administration, the combination, composition, or kit may contain one or more sweeteners, preservatives, colorants, and flavor enhancers. In combinations or compositions intended for administration by injection, one or more surfactants, preservatives, wetting agents, dispersants, suspending agents, buffers, stabilizers, and isotonic agents may also be included. In the kit of the present invention, the liquid components containing one or more of the active agents of the composition may be combined and administered simultaneously before administration, or each active agent may be administered sequentially or separately.

[0130] The preferred route of administration is parenteral administration, including, but is not limited to, intradermal, intramuscular, intraperitoneal, intravenous, subcutaneous, intranasal, epidural, intracerebral, intraventricular, intrathecal, vaginal, or percutaneous. The preferred mode of administration is left to the discretion of the practitioner and, in some cases, will depend on the location of the medical condition (e.g., the location of cancer).

[0131] In a more preferred embodiment, the combination, composition and kit of the present invention are administered intravenously.

[0132] The liquid combinations, compositions, and kits of the present invention, whether they are solutions, suspensions, or other similar forms, may also include one or more of the following: sterile diluents such as water for injection, saline solution, preferably physiological saline, Ringer's solution, isotonic sodium chloride, non-volatile oils such as synthetic mono or diglycerides, polyethylene glycol, glycerin, or other solvents; antimicrobial agents such as benzyl alcohol or methylparaben; and agents for adjusting tonicity such as sodium chloride or dextrose. Parenteral combinations or compositions may be sealed in ampoules, disposable syringes, or multiple dose vials made from glass, plastic, or other materials. Physiological saline is a preferred adjuvant.

[0133] For administration (e.g., intravenously), the combination, composition, or kit typically contains 10-50 mg of tinostamstine or a pharmaceutically acceptable salt thereof per patient's body surface area m². 2 Within the dosage range, patients can choose to receive 20-40 mg / m² of their body surface area. 2 Within this dosage range, preferably 25-40 mg / patient body surface area m². 2 Preferably, an additional 25-35 mg / patient body surface area m² 2 (For example, 30 mg / m²) 2 ) within the dosage range, and immune checkpoint inhibitors at 1-5 mg / patient body surface area m 2 Within this dosage range, preferably 2-4 mg / patient body surface area m² 2 Preferably, an additional approximately 3 mg / patient body surface area m² 2 (For example, 3 mg / m²) 2 It may be included within the dosage range of )

[0134] The combinations, compositions, or kits of the present invention can be formulated so that an immune checkpoint inhibitor and tinostamustine or a pharmaceutically acceptable salt thereof are administered simultaneously, sequentially, or separately. Preferably, they are administered separately.

[0135] The combinations, compositions, or kits of the present invention can be administered by any convenient route, for example, by injection or large-volume injection, or by absorption through the epithelium or mucocutaneous lining.

[0136] In specific embodiments, it may be desirable to administer one or more combinations, compositions, or kits of the present invention topically to an area requiring treatment. In one embodiment, administration is obtained by direct injection to a site (or previous site) of cancer, tumor, neoplasm, or pre-neoplastic tissue.

[0137] Lung administration may also be used, for example, by the use of an inhaler or nebulizer and a formulation containing an aerosolizing agent, or by perfusion in fluorocarbon or synthetic alveolar surfactant. In certain embodiments, the combination, composition or kit or composition of the present invention may be formulated as a suppository using a conventional binder and carrier such as a triglyceride.

[0138] This combination, composition, or kit may take the form of a solution, suspension, emulsion, tablet, pill, pellet, capsule, liquid-containing capsule, powder, sustained-release formulation, suppository, emulsion, aerosol, spray, suspension, or any other form suitable for use. Other examples of suitable pharmaceutical carriers are described in "Remington's Pharmaceutical Sciences" by E.W. Martin.

[0139] Pharmaceutical combinations, compositions, and kits can be prepared using methods well known in the pharmaceutical field. For example, a composition intended to be administered by injection can be prepared by combining the components of the kit of the present invention with water to form a solution. Surfactants may be added to facilitate the formation of a homogeneous solution or suspension.

[0140] The combinations, compositions, and kits of the present invention are particularly effective in the treatment of cancer. As described in more detail in the examples below, the claimed combinations of the present invention have been found to be more effective in the treatment of cancer than current standard treatments.

[0141] According to an eighth aspect of the present invention, the use of a combination comprising tinostamustine, or a pharmaceutically acceptable salt thereof, and an immune checkpoint inhibitor in the treatment of cancer is provided. Cancer and / or said treatment may be as defined by the aspects of the present invention described above.

[0142] According to the ninth aspect, the present invention also covers the use of combinations, compositions, or kits according to the first, second, or third aspects of the present invention in the manufacture of pharmaceuticals for treating cancer. Cancer and / or the treatment may be as defined by any of the aspects of the present invention described above.

[0143] According to a tenth aspect of the present invention, the use of an immune checkpoint inhibitor in the manufacture of a pharmacopoeia for use in the treatment of cancer is provided, wherein in the treatment, the immune checkpoint inhibitor is used in combination with tinostamstine or a pharmaceutically acceptable salt thereof. Cancer and / or the treatment may be as defined by any of the aspects of the present invention described above.

[0144] According to an eleventh aspect of the present invention, tinostamustine or a pharmaceutically acceptable salt thereof is provided for use in the treatment of melanoma, optionally for use in the treatment of progressive melanoma, and optionally for use in the treatment of recurrent and / or refractory melanoma. Cancer and / or said treatment may be as defined by any of the aspects of the present invention described above.

[0145] According to a twelfth aspect of the present invention, tinostamustine, or a pharmaceutically acceptable salt thereof, is provided for use in the treatment of colorectal cancer, in which the colorectal cancer is optionally the carcinoma.

[0146] As defined, the treatment method, which involves administering to the cancer being treated and the patient a therapeutic dose of tinostammustine, or a pharmaceutically acceptable salt thereof, or a therapeutically effective dose of an immune checkpoint inhibitor, will be found to be equally applicable to all aspects of the present invention as defined herein. [Examples]

[0147] (Example 1) The examples below relate to the final clinical results from clinical trial registration numbers National Clinical Trials (NCT) 030903458 and Swiss National Clinical Trials Portal (SNCTP) 000003243, which are ongoing.

[0148] The clinical trial concerns an open-label, single-center, Phase 1B study to determine the safety, tolerability, recommended dose, and anticancer efficacy of tinostamustine in combination with the anti-PD-1 monoclonal antibody (mAb), nivolumab, in patients with advanced melanoma receiving standard treatment. Tinostamustine has the following structural formula:

[0149] [ka]

[0150] It is a compound that has [a certain characteristic].

[0151] Guidance on current standard treatment for patients with metastatic melanoma can be found in Keilholz et al. (doi: https: / / doi.org / 10.1016 / j.annonc.2020.07.004). As those skilled in the art will recognize, the treatment given to a particular patient depends on the individual patient's circumstances.

[0152] material Tinostamustine was supplied by Mundipharma Research Limited (Cambridge, UK) as 100 mg of lyophilized powder for reconstitution and intravenous administration. Nivolumab was supplied by Bristol-Myers-Squibb (BMS) as a 100 mg / 10 mL concentrate for intravenous administration (optionally diluted).

[0153] Important selection criteria Patients with inoperable stage III or metastatic stage IV melanoma confirmed by histological or cytological means; The patient is (i) not being considered for first-line combination anti-PD(L)1 / anti-CTLA4 immunotherapy, or (ii) has failed standard palliative systemic treatment including anti-PD(L)1 / anti-CTLA4 immunotherapy and / or BRAF / MEK-targeted tyrosine kinase inhibitors; • Performance status of the East Coast Cancer Clinical Group (ECOG) in the United States is ≤2. Patients with brain metastases should have undergone curative treatment (surgery or radiation therapy) at least two weeks prior to initiating the study drug and should be documented as having a stable disease on imaging; • Appropriate bone marrow, kidney, and liver function; • Appropriate contraception.

[0154] Important Exclusion Criteria • Patients prior to adjuvant treatment with PD(L)1-targeted monoclonal antibodies, except for those who completed adjuvant PD(L)1-targeted treatment at least 6 months prior to the start of the investigational treatment; • Patients who discontinued prior adjuvant anti-PD(L)-1 treatment due to anti-PD(L)-1 related toxicity; • Patients who have received systemic treatment or radiation therapy within two weeks prior to starting the investigational drug; • Concomitant treatment with a systemic steroid at a daily dose equivalent to ≥10 mg of prednisone, or with an immunosuppressant such as methotrexate; Patients with a preceding malignant lesion (excluding non-melanoma skin cancer and carcinoma in situ such as bladder, colon, cervical / dysplasia, melanoma, or breast cancer). Patients with another secondary malignant lesion diagnosed two years prior to the present, who have been treated with the intention of curing, have had no evidence of disease during that period, and who the principal investigator has determined to have a low risk of recurrence are eligible; • New York Heart Association (NYHA) Stage III / IV congestive heart failure and / or arrhythmias are not adequately controlled; • Corrected QT (QTc) interval (Friderician formula) > 450 msec • Patients treated with drugs known to prolong the QT / QTc interval (Credible Meds list: Known risk of TdP: https: / / crediblemeds.org / new-drug-list) • Pregnant and breastfeeding patients.

[0155] Patient cohort The clinical trial recruited patients with advanced melanoma based solely on the exclusion and selection criteria described above. Melanoma has generally been shown to respond to immunotherapy, particularly immune checkpoint inhibitors. Therefore, the trial focused on this homogeneous patient population to gain insight into the clinical activity of the combination of tinostamustine and an immune checkpoint inhibitor, namely nivolumab, in patients with advanced melanoma. However, those skilled in the art will recognize that the scope of the present invention is not limited to the treatment of melanoma and that the combination described herein may be equally used for the treatment of any cancer.

[0156] A total of 17 patients with advanced melanoma were recruited to a Phase IB trial to characterize the safety, tolerability, recommended dose, and preliminary efficacy of tinostamustine in combination with the immune checkpoint inhibitor nivolumab for the treatment of cancer.

[0157] Thirteen patients (77%) received treatment with immune checkpoint inhibitors as an initial treatment line, and of those, 11 patients (64%) received treatment with immune checkpoint inhibitors as first-line treatment.

[0158] Ten patients (59%) had elevated lactate dehydrokinase (LDH) baselines. Lactate dehydrokinase (LDH) levels were inversely correlated with the response to checkpoint inhibitors. High LDH levels are products of enhanced glycolytic activity in tumors and hypoxic tumor necrosis, the latter being associated with high tumor burden (Van Wilpe S, Koornstra R, Den Brok M, De Groot JW, Blank C, De Vries J, Gerritsen W, Mehra N. Lactate dehydrogenase: a marker of diminished antitumor immunity. Oncoimmunology. 2020 Feb 26;9(1):1731942. doi: 10.1080 / 2162402X.2020.1731942. PMID: 32158624; PMCID: PMC7051189).

[0159] Four patients (ENI-SG-03, ENI-SG-01, ENI-SG-10, ENI-SG-15) were treatment-naive, and ten patients had received at least one line of treatment (ENI-SG-13, ENI-SG-06, ENI-SG-16, ENI-SG-05, ENI-SG-14, ENI-GR-04, ENI-SG-11, ENI-SG-12, ENI-SG-08, ENI-GR-17, ENI-SG-09, ENI-SG-07). Seven patients had melanoma phenotypes associated with poor prognosis, including acral lentiginous melanoma (3 patients), mucosal melanoma (1 patient), and uveal melanoma (3 patients). Other melanoma phenotypes included nodular melanoma (3 patients) and superficial spreading melanoma (2 patients).

[0160] Dosage The dosage of tinostamstine is expressed herein as the patient's surface area, i.e., mg / m². 2 It is defined by reference to the amount of tinostamustine used for [the patient]. An experienced clinician can use common sense to calculate the patient's surface area (also referred to herein as body surface area). In particular, the patient's surface area (PSA) can be calculated using the following formula (Dubois D, Dubois EF, A formula to estimate the approximate surface area if height and weight be known, Arch Intern Med, 1916, 17, pp. 863-871): PSA = 0.007184 × (patient's height in cm) 0.725 × (Patient's weight in kg) 0.425

[0161] Tinostamustine was administered intravenously. Four patients received 15 mg / m² of tinostamustine per cycle. 2 The drug was administered at a dose (referred to as dose level 1-DL1). Thirteen patients received tinostamstine at 30 mg / m² per cycle. 2 It was administered at the dose (referred to as dose level 2-DL2).

[0162] Nivolumab was administered intravenously. All patients received nivolumab at a dose of 3 mg / kg of patient body weight per cycle, with the exception of cycle 1 (C1), as outlined below. The nivolumab dose was calculated according to the patient's body weight and was based on clinical recommendations for the use of nivolumab (NCT01721772; Topalian et al., N Engl J Med 366: pp. 2443-2454, 2012).

[0163] Patient weight and BSA were determined within two days prior to the start of each new treatment cycle.

[0164] Dosage plan The treatment was administered in 14-day cycles. A total of 36 cycles were planned for the experimental portion.

[0165] Cycle 1 (C1) - On day 1 (D1) of Cycle 1 (C1), only tinostamustine was administered to the patient at the prescribed dose; nivolumab was not administered to the patient. Tinostamustine was administered intravenously by infusion over 1 hour.

[0166] Second cycle (C2) - On day 1 (D1) of the second cycle (C2), nivolumab was administered intravenously to the patient at a dose of 3 mg / kg over 1 hour by infusion. Tinostamustine was administered to the patient at the prescribed dose at least 30 minutes after the completion of nivolumab administration. Tinostamustine was administered intravenously by infusion over 1 hour.

[0167] For all subsequent cycles – after C2, nivolumab was administered intravenously to the patient at a dose of 3 mg / kg by infusion over 30 minutes on day 1 (D1) of each subsequent cycle (Cn; n is comprehensively selected from a consecutive integer between 3 and 36, depending on the total number of cycles the patient is participating in the study). Tinostamustine was administered to the patient at the prescribed dose at least 30 minutes after the completion of nivolumab administration. Tinostamustine was administered intravenously by infusion over 1 hour.

[0168] Therefore, nivolumab and tinostamustine were administered to the patients every two weeks (i.e., starting with cycle 2 and on day 1 of each cycle).

[0169] Evaluation items Treatment was continued until the first occurrence of any of the following: progressive disease, unacceptable toxicity, withdrawal of informed consent, or the investigator's judgment. Patients with progressive disease could continue the trial provided that clinical benefit continued.

[0170] The primary endpoint of the study was dose-limiting toxicity (DLT) observed during the first six weeks of the study treatment.

[0171] The secondary evaluation criteria for the exam include: • Objective tumor response (OR) to the investigational treatment, as measured by Response Evaluation Criteria in Solid Tumors (RECIST) v1.1 and iRECIST; • All adverse events (AEs) according to the Common Terminology Criteria for Adverse Events (CTCAE); • Progression-free survival (PFS) is defined as the time from enrollment in the trial until the first occurrence of disease progression or patient death. · Overall survival (OS) It included.

[0172] Objective tumor response (OR) may be defined as complete response (CR) or partial response (PR) during the study treatment. Progression-free survival (PFS) may be defined as the time from enrollment in the study to the first occurrence of disease progression (PD) or patient death. Overall survival (OS) is defined as the length of time from the start of the study treatment to death for any reason.

[0173] Patients were followed up until at least 100 days after the last experimental treatment with nivolumab, or at least 30 days after the last experimental treatment with tinostamustine, whichever was later. Patients were also followed up to assess disease progression and overall survival.

[0174] result The following data reports the number of treatment cycles received, as well as the PFS and OS days for each mobilized patient.

[0175] [Table 1]

[0176] Three patients withdrew from the trial in the early stages due to progressive disease. Fourteen patients continued the trial, forming the trial cohort, as outlined in more detail below.

[0177] The safety part of the study was 10 mg / m² of body surface area. 2 From 30 mg / m² of body surface area 2 The administration of tinostamustine at doses between these levels progressed without any significant safety concerns being identified. Therefore, the recommended dose of tinostamustine is 30 mg / m². 2 It was decided that this was the case.

[0178] In the efficacy part of the trial, the mean number of cycles for all patients was 10.8 (22 weeks). The median progression-free survival (mPFS) for all patients was 1.94 (95% CI 1.55, 7.89) months. The median overall survival (mOS) for all patients was 9.27 (95% CI 1.97, 22.29) months. One treatment-related serious adverse event (SAE), nivolumab-associated immune-associated pneumonitis (ENI-GR-04), was observed. No other patients were determined to have treatment-related adverse events.

[0179] Three patients (ENI-SG-02, ENI-SG-07, and ENI-SG-09) received less than four weeks or equivalent (i.e., less than two cycles) of the study treatment due to disease progression in the early stages of the trial.

[0180] Of the remaining 14 patients, stable disease was observed in 3 patients (21.4%; ENI-SG-01, ENI-GR-04, ENI-SG-06). Partial response was observed in 3 patients (21.4%; ENI-SG-03, ENI-SG-13, ENI-SG-16).

[0181] Therefore, seven patients (ENI-SG-1, ENI-SG-3, ENI-GR-4, ENI-SG-6, ENI-SG-13, ENI-SG-15, and ENI-SG-16; shaded in Table 1 above) were considered to have a clinical response to treatment, defined as at least 60 days without disease progression (PFS = ≥ 60 days). Considering only these patients who were considered to have a clinical response to treatment, the median progression-free survival (mPFS) increased to 7.04 months and the median overall survival (mOS) increased to 10.73 months.

[0182] Three patients (ENI-SG-03, ENI-GR-04, and ENI-GR-17) were alive at the time of writing. Of these three patients, two (ENI-SG-13 and ENI-SG-16) were alive at the time of writing without developing progressive disease (PD).

[0183] The mPFS and mOS data outlined above were compared with previously published reference data describing mPFS and mOS with the current standard of care for advanced melanoma, namely the combination of nivolumab and ipilimumab. The references outlined below, along with the mPFS and mOS data, are considered by those skilled in the art to be representative of the predicted clinical outcomes in patients with advanced melanoma treated with the combination of nivolumab and ipilimumab as a second line of treatment.

[0184] One trial reported a mPFS of 2 months in patients with advanced melanoma treated with a combination of nivolumab and ipilimumab as a second line of treatment (Zimmer L et al.; Ipilimumab alone or in combination with nivolumab after progression on anti-PD-1 therapy in advanced melanoma. Eur J Cancer. 2017 Apr;75:47~55. doi: 10.1016 / j.ejca.2017.01.009. Epub 2017 Feb 17. PMID: 28214657).

[0185] Another study reported a mOS of 5.6 months in patients with advanced melanoma treated with a combination of nivolumab and ipilimumab as a second line of treatment (Baron K et al.; Comparative effectiveness of second-line ipilimumab vs. nivolumab in combination with ipilimumab in patients with advanced melanoma who received frontline anti-PD-1 antibodies. Journal of Oncology Pharmacy Practice. 2021;27(3):555~559. doi:10.1177 / 1078155220924719).

[0186] The median progression-free survival and median overall survival for patients in the trials outlined above can be summarized below (Table 2):

[0187] [Table 2]

[0188] Figure 1 shows the PFS for all patients mobilized in the trial (see Table 1 above), and further includes comparisons of mPFS for all patients, mPFS for patients considered to have responded to the treatment (PFS = ≥ 60 days), and mPFS reported by Zimmer et al. (see reference provided above; see Table 2). These data suggest that the nivolumab and tinostamustine combination increased mPFS by 8.95 months in patients considered to have responded to the treatment (PFS = ≥ 60 days) compared to the current absolute standard of care, the nivolumab and ipilimumab combination.

[0189] Figure 2 shows the overall survival (OS) for all patients mobilized in the trial (see Table 1 above), and further includes comparisons of the overall life-sustaining (mOS) for all patients, mOS (PFS ≥ 60 days) for patients considered to have responded to the treatment, and mOS reported by Baron et al. (see reference above). These data suggest that the nivolumab and tinostamustine combination increased mOS by 3.87 months compared to the current absolute standard of care, the nivolumab and ipilimumab combination, and increased mOS by 21.07 months in patients considered to have responded to the treatment (PFS ≥ 60 days).

[0190] These data demonstrate for the first time that the combination of tinostamustine and nivolumab is an effective treatment for cancer, particularly for advanced melanoma. Remarkably, the combination of tinostamustine and nivolumab has been shown to increase PFS and OS compared to current standard treatments, and is therefore found to be a promising treatment for use in cancer treatment. Those skilled in the art will recognize that nivolumab is approved for use in the treatment of multiple forms of cancer, and that the remarkably effective combination of tinostamustine and nivolumab, as described herein, can for the first time be used in the treatment of any cancer.

[0191] (Example 2) The inventors have tested the in vivo efficacy of tinostamustine and immune checkpoint inhibitors in the treatment of colorectal cancer in mice and found them to be effective. Specifically, the effects of combined therapy with tinostamustine and an anti-PLD1 antibody in a subcutaneous mouse colorectal allograft model of female C57BL / 6 mice were evaluated after three dose cycles. MC38 is a mouse model commonly used for colorectal cancer.

[0192] The mice from the six groups were randomly assigned to groups 1 through 6, as shown in Table 3.

[0193] [Table 3]

[0194] The inventors demonstrate in Figures 3 to 6 that a treatment regimen of tinostamustine and immune checkpoint inhibitors is effective against colorectal cancer in vivo. Specifically, a treatment regimen of tinostamustine and PD-1 inhibitors (anti-mouse PD1, RMP1-14, Bioxcell) was effective in treating a B16-F10 subcutaneous mouse melanoma allograft model in female C57BL / 6 mice.

[0195] Figure 3 shows that tinostamustine monotherapy or combination therapy with an anti-PD1 inhibitor was well-tolerated in mice and did not result in weight loss. In fact, the mean body weight of mice during treatment was observed in both the monotherapy and combination therapy groups. Combination therapy (groups 5 and 6) achieved a significant reduction in tumor volume compared to the control group (group 1) and the group treated with tinostamustine monotherapy.

[0196] Supporting this, Figure 6 shows that the claimed combination therapy significantly increased the mean survival time of mice compared to vehicle controls and tinostamustine monotherapy.

[0197] In summary, compared to the vehicle group, the combination treatments (Group 5: tinostamustine 5 mg / kg and anti-PD1 10 mg / kg, Group 6: tinostamustine 15 mg / kg and anti-PD1 10 mg / kg) showed significant antitumor efficacy. Furthermore, the combination treatment groups showed additive effects in the MC38 model. Tinostamustine monotherapy and tinostamustine combined with an anti-PD1 inhibitor significantly extended the survival time of mice in the MC38 model. In this study, the mice tolerated the drugs well, and there was no significant weight loss during the study.

[0198] Methods and materials Dosage According to Example 2, the dosage of combination therapy may be 5 mg / kg or 15 mg / kg of tinostamustine and 10 mg / kg of a PD-1 inhibitor. According to Example 2, the preferred dosage schedule is tinostamustine once a week and the PD-1 inhibitor once every two weeks.

[0199] Tinostamustine was administered intravenously, specifically by slow intravenous injection at a rate of approximately 60 seconds per mouse. Two groups of mice were administered tinostamustine at a dose of 5 mg / kg once a week, either as monotherapy (group 3) or in combination with other therapies (group 5). The other two groups of mice were administered tinostamustine at a dose of 15 mg / kg once a week, either as monotherapy (group 4) or in combination with other therapies (group 6).

[0200] The anti-PDL1 antibody (RMP1-14) was administered by intraperitoneal injection to the combination therapy groups (groups 4 and 6). Mice in groups 4 and 6 received the anti-PDL1 antibody at a dose of 10 mg / kg of mouse body weight once every two weeks.

[0201] cell culture MC38 cells were maintained at 37°C in DMEM medium supplemented with 10% fetal bovine serum under an atmosphere of 5% CO2 in air. Cells in the exponential growth phase were collected and quantified using a cell counter before tumor inoculation.

[0202] Tumor Inoculation Each mouse was subcutaneously inoculated with MC38 tumor cells (1 × 10⁶) in 0.1 ml of PBS in the right lower flank region to induce tumor development.

[0203] Randomization Randomization was initiated when the average tumor size reached approximately 79 mm³. Ninety mice were assigned to the study, and all animals were randomly assigned to six test groups, with 15 mice in each group.

[0204] Administration of test substance The procedure was initiated on day 0 for each test design.

[0205] Observation and data collection After tumor cell inoculation, animals were checked daily for disease and death. During routine monitoring, animals were checked for tumor growth, behavioral changes such as motility, food and water consumption, weight gain / loss (weight was measured twice a week after randomization), eye / hairball and any other abnormalities, and the effect of treatment. Deaths and observed clinical signs were recorded in detail for each individual animal.

[0206] Tumor volume was measured twice weekly in two dimensions using calipers after randomization, and the volume was expressed in mm³ using the formula V = (L × W × W) / 2, where V is the tumor volume, L is the tumor length (longest tumor dimension), and W is the tumor width (longest tumor dimension perpendicular to L). Medication, as well as tumor and body weight measurements, were performed using a Laminar Flow Cabinet. Body weight and tumor volume were measured using StudyDirector™ software (version 3.1.399.19).

[0207] End of exam The procedure was performed over 20 days (days 0-19). The trial concluded on day 19.

Claims

1. A combination comprising tinostamustine, or a pharmaceutically acceptable salt thereof, and an immune checkpoint inhibitor.

2. The combination according to claim 1, wherein the immune checkpoint inhibitor is a programmed cell death protein-1 (PD-1) inhibitor.

3. The combination according to claim 2, wherein the immune checkpoint inhibitor is an anti-PD-1 antibody.

4. The combination according to claim 3, wherein the immune checkpoint inhibitor is selected from pembrolizumab, nivolumab, dostallimab, retifanlimab, and semiprimab, and optionally the immune checkpoint inhibitor is nivolumab.

5. The combination according to claim 1, wherein the immune checkpoint inhibitor is a programmed cell death protein ligand 1 (PD-L1) inhibitor and / or a programmed cell death protein ligand 2 (PD-L2) inhibitor, and optionally the immune checkpoint inhibitor is a PD-L1 inhibitor.

6. The combination according to claim 5, wherein the immune checkpoint inhibitor is a PD-L1 inhibitor that is an anti-PD-L1 antibody, and is optionally selected from atezolizumab, avelumab, and durvalumab.

7. The combination according to any one of claims 1 to 6, further comprising a cytotoxic T lymphocyte-associated antigen 4 (CTLA-4) inhibitor, wherein the CTLA-4 inhibitor is optionally selected from ipilimumab and tremelimumab, and optionally further comprising ipilimumab.

8. A pharmaceutical composition comprising a combination according to any one of claims 1 to 7 and a pharmaceutically acceptable diluent or carrier.

9. A kit comprising a combination according to any one of claims 1 to 7 or the pharmaceutical composition according to claim 8, and optionally, instructions for treating a patient.

10. A combination according to any one of claims 1 to 7, a pharmaceutical composition according to claim 8, or a kit according to claim 9 for use in the treatment of cancer.

11. A combination, pharmaceutical composition, or kit for use according to claim 10, wherein the cancer is selected from melanoma, small cell lung cancer, non-small cell lung cancer, mesothelioma (e.g., malignant pleural mesothelioma), renal cell carcinoma, head and neck cancer, urothelial carcinoma, colorectal cancer, esophageal squamous cell carcinoma, hepatocellular carcinoma, gastric cancer, esophageal cancer, esophageal adenocarcinoma, gastroesophageal junction cancer, high-frequency microsatellite instability or mismatch repair deficiency cancer, primary mediastinal large B-cell lymphoma (PMBCL), Merkel cell carcinoma (MCC), endometrial cancer, high tumor mutational load (TMB-H) cancer, cutaneous squamous cell carcinoma (cSCC), soft tissue sarcoma, osteosarcoma, ovarian cancer, trinegative breast cancer, glioblastoma, multiple myeloma, Hodgkin lymphoma, and non-Hodgkin lymphoma.

12. The combination, pharmaceutical composition, or kit for use according to claim 11, wherein the cancer is, optionally, melanoma, or progressive melanoma.

13. A combination, pharmaceutical composition, or kit for use according to any one of claims 10 to 12, wherein the cancer is recurrent and / or refractory.

14. A combination, pharmaceutical composition, or kit for use according to any one of claims 10 to 13, wherein in the treatment described above, tinostamustine is administered to patients in need as an adjuvant and / or neoadjuvant, and optionally, tinostamustine is administered to patients in need as a neoadjuvant.

15. In the aforementioned treatment, tinostamustine, or a pharmaceutically acceptable salt thereof, is administered to the patient at a dose of 10-50 mg / m² of body surface area. 2 Optionally, 20-40 mg / m² 2 Additionally, an optional additional 25-35 mg / m² 2 The dosage range is as follows, with an optional 30 mg / m². 2 A combination, pharmaceutical composition, or kit for use according to any one of claims 10 to 14, administered by...

16. A combination, pharmaceutical composition, or kit for use according to any one of claims 10 to 15, wherein the treatment involves administering tinostamstine, or a pharmaceutically acceptable salt thereof, and the immune checkpoint inhibitor to the patient sequentially, simultaneously, or separately.

17. The combination, pharmaceutical composition, or kit according to claim 16, comprising administering tinostamustine, or a pharmaceutically acceptable salt thereof, and the immune checkpoint inhibitor separately, and optionally administering the immune checkpoint inhibitor first to a patient in need, followed by the administration of tinostamustine.

18. The combination, pharmaceutical composition, or kit according to claim 17, wherein tinostamstine is administered to a patient in need for 20 to 120 minutes, optionally for 30 to 90 minutes, and optionally for a further 50 to 70 minutes.

19. A combination, pharmaceutical composition, or kit for use according to any one of claims 10 to 18, wherein in the first cycle of treatment, tinostamustine, or a pharmaceutically acceptable salt thereof, is administered to the patient in need, without the checkpoint inhibitor, and in all subsequent cycles, tinostamustine and / or a pharmaceutically acceptable salt thereof, and the immune checkpoint inhibitor are administered to the patient successively, simultaneously, or separately, preferably separately.

20. Tinostamustine, or a pharmaceutically acceptable salt thereof, for use in the treatment of cancer, in combination with an immune checkpoint inhibitor.

21. The cancer is as defined in any one of claims 11 to 13, and / or the treatment is as defined in any one of claims 14 to 19, tinostamstine for use according to claim 20, or a pharmaceutically acceptable salt thereof.

22. An immune checkpoint inhibitor for use in the treatment of cancer, used in combination with tinostamustine or a pharmaceutically acceptable salt thereof.

23. An immune checkpoint inhibitor for use according to claim 22, wherein the cancer is as defined in any one of claims 11 to 13, and / or the treatment is as defined in any one of claims 14 to 19.

24. A method for treating cancer, comprising administering to a patient in need of such treatment a therapeutically effective dose of tinostamustine, or a pharmaceutically acceptable salt thereof, in combination with a therapeutically effective dose of an immune checkpoint inhibitor.

25. The use of tinostamustine, or a pharmaceutically acceptable salt thereof, and a combination including immune checkpoint inhibitors in the treatment of cancer.

26. Use of tinostamustine or a pharmaceutically acceptable salt thereof in the manufacture of a pharmaceutical product for use in the treatment of cancer, wherein the tinostamustine or a pharmaceutically acceptable salt thereof is used in combination with an immune checkpoint inhibitor in the treatment.

27. Use of an immune checkpoint inhibitor in the manufacture of a pharmaceutical product for use in the treatment of cancer, wherein the immune checkpoint inhibitor is used in combination with tinostamstine or a pharmaceutically acceptable salt thereof in the treatment.

28. The method according to claim 24, the use according to claim 25, or the use according to claim 26 or claim 27, wherein the cancer is as defined in any one of claims 11 to 13, and / or the treatment is as defined in any one of claims 14 to 19.

29. Tinostamustine, or pharmaceutically acceptable salt thereof, for use in the treatment of melanoma, optionally for use in the treatment of progressive melanoma, and further optionally for use in the treatment of recurrent and / or refractory melanoma.

Images