Use of sodium trans-[tetrachloridobis(1h-indazole)ruthenate(III)] for treating TP53 mutant cancers

Abstract

Methods and corresponding uses are provided for treating a cancer having a TP53 mutation in a patient in need thereof, comprising administering an effective amount of sodium trans-[tetrachloridobis(1H-indazole)ruthenate(III)] (BOLD-100). BOLD-100 may be used in combination with an effective amount of an additional chemotherapeutic agent.

Description

USE OF SODIUM TRANS-[TETRACHLORIDOBIS(1H-INDAZOLE)RUTHENATE(III)] FOR TREATING TP53 MUTANT CANCERS FIELD

[0001] The invention is in the field of therapeutics, including the use of sodium trans- [tetrachloridobis(1H-indazole)ruthenate(III)] for treating cancers bearing mutations in the TP53 gene. BACKGROUND

[0002] Sodium trans-[tetrachloridobis(1H-indazole)ruthenate(III)] is a coordinated complex of ruthenium having anticancer activity (also known as BOLD-100, KP1339, NKP-1339, IT-139, and Na[RuIIICl4(Hind)2]). Methods of making alkali metal salts of trans-[tetrachlorobis(1H- indazole)ruthenate (III)] are for example described in PCT Patent Publication WO2018204930, such compounds having Formula I:wherein M is an alkali metal cation, including the sodium salt:

[0003] The TP53 gene in humans is a tumour suppressor gene encoding p53 proteins (p53 isoforms a through l) having roles in regulating progression of the cell cycle, apoptosis, and genomic stability (also known as tumor protein P53, cellular tumor antigen p53 (UniProt name), or transformation-related protein 53 (TRP53)). The TP53 gene is identified by NCBI Gene ID 7157, on chromosome 17 at map location 17p13.1, and its canonical transcript is identified as NM_000546. TP53 is commonly referred to as the “guardian of the genome” reflecting its role as a crucial tumour suppressor involved in regulating cell division and preventing tumor development. Mutations in TP53 are some of the most common genetic alterations in a wide variety of cancers (see Chen, X., Zhang, T., Su, W. et al., Mutant p53 in cancer: from molecular mechanism to therapeutic modulation. Cell Death Dis 13, 974 (2022)). The Clinical Knowledgebase (CKB) managed by The Jackson Laboratory as a resource for interpreting complex cancer genomic profiles lists hundreds of known variants of the TP53 gene, ascribing effects to many of them. TP53 mutations are often prognostic for worse overall survival (OS) in a range of cancers. Mutations can occur throughout the TP53 gene and can be separated into gain-of-function (GOF) and non-GOF mutations. GOF mutations are those that impact additional anticancer functions in addition to the standard loss of TP53 function. GOF mutations are commonly prognostic for worse OS in a range of cancers, although this varies with the type of cancer (see Ottaiano et al. The prognostic role of p53 mutations in metastatic colorectal cancer: A systematic review and meta-analysis. Crit Rev Oncol Hematol.2023 Jun;186:104018). SUMMARY

[0004] Methods and corresponding uses are provided for treating a cancer characterized by a TP53 mutation (TP53MT) in patients in need thereof, such as a human patient, comprising administering an effective amount of sodium trans-[tetrachloridobis(1H- indazole)ruthenate(III)] (BOLD-100). BOLD-100 may be used alone or in combination with another therapeutic agent. The cancer may be a cancer that is resistant to treatment with BOLD-100 alone or is a cancer that is resistant to treatment with another chemotherapy agent or chemotherapy regimen, and may for example be a metastatic cancer. The cancer may be a colorectal cancer (CRC), such as a CRC adenocarcinoma.

[0005] One general aspect includes a method of treating a cancer with a mutation in a TP53 gene in a human patient in need thereof. The method may also include identifying thecancer in the patient as having the TP53 mutation. The method involves administering an effective amount of sodium trans-[tetrachloridobis(1h-indazole)ruthenate(III)] to treat the cancer.

[0006] Implementations may include one or more of the following features. The method where the TP53 mutation mediates a substitution, deletion or insertion at an altered amino acid position in a p53 protein encoded by the TP53 gene. The altered amino acid position may for example be at one or more of positions 175, 245, 248, 249, 273, or 282 in the sequence set out in SEQ ID NO:2. The tp53 mutation mediates a substitution in the p53 protein that may for example be one or more of R175A, R175C, R175G, R175H, R175L R175P, G245A, G245C, G245D, G245F, G245R, G245S, G245V, R248E, R248G, R248L, R248P, R248Q, R248W, R249G, R249K, R249M, R249S, R249T, R249W, R273C, R273H, R273L, R273P, R273S, R273W, R273Y, R282D, R282G, R282H, R282Q or R282W. The mutation may for example be a gain-of-function (GOF) mutation in the TP53 gene. The GOF mutation mediates a substitution in the p53 protein that may for example be one or more of S121X, C124X, V157X, A161X, R175X, L194X, T211X, F212X, H214X, G245X, R248X, R249X, R273X, C277X, R280X, D281X, R282X, R290X, K291X, K292X, or G360X, where x is any substitute amino acid. The substitution in the p53 protein may for example be one or more of S121A, S121C, S121F, C124S, V157S, A161T, R175H, L194R, T211A, F212Y, H214R, G245A, G245C, G245D, G245S, R248G, R249W, R273H, C277S, C277Y, R280K, D281G, R282Q, R290G, K291E, K291Q, K291T, K291I, K292T, or G360V. The cancer may for example be a colorectal, gastric, pancreatic, esophageal, breast or biliary tract cancer. The cancer may for example be a metastatic colorectal cancer (mCRC). The cancer may for example be a stage II, III or IV cancer. Sodium trans-[tetrachloridobis(1h-indazole)ruthenate(III)] may for example be administered in combination with one or more additional therapeutic agent to treat the cancer. The additional therapeutic agent may include a platinum-based chemotherapeutic; an antimetabolite; an anthracycline; an EGFR targeted therapeutic; a poly (ADP-ribose) polymerase parp targeted therapeutic; a vascular endothelial growth factor (VEGF) or vascular endothelial growth factor receptor (VEGFR) targeted therapeutic, or an immunotherapeutic. The platinum-based chemotherapeutic may include cisplatin, carboplatin or oxaliplatin. The antimetabolite may include methotrexate, 5-FU, aminopterin, cytarabine, thioguanine, or mercaptopurine. The anthracycline may include daunorubicin, doxorubicin, epirubicin, idarubicin, mitoxantrone or valrubicin. The EGFR targeted therapeutic may include afatinib, gefitnib, amivantamab-vmjw, osimertinib, erlotinib, or dacomitinib. The parp targeted therapeuticolaparib, rucaparib, or niraparib. VEGF / VEGFR targeted therapeutic may include sunitinib, pazopanib, foretinib, sorafenib, tivozanib, anlotinib, axitinib, golvatinib, fresolimumab, trastuzumab, ramucirumab, tanibirumab, sym004, bevacizumab, aflibercept, or sevacizumab. The cancer may for example be a colorectal cancer (CRC) or metastatic colorectal cancer (mCRC) and sodium trans-[tetrachlorobis(1h-indazole)ruthenate (III)] may for example be administered in combination with a FOLFOX treatment, such as an mFOFOX6 regimen. The cancer may for example be a colorectal cancer (CRC) or metastatic colorectal cancer (mCRC) and sodium trans-[tetrachlorobis(1h-indazole)ruthenate (III)] may for example be administered in combination with irinotecan. The cancer may for example be a colorectal cancer (CRC) or metastatic colorectal cancer (mCRC) and sodium trans-[tetrachlorobis(1h-indazole)ruthenate (III)] may for example be administered in combination with a VEGF inhibitor, an EGFR inhibitor, a PARP inhibitor. The treatment may for example be a first line treatment. The treatment may for example be second, third, fourth, or subsequent line treatment. The treatment may for example be an adjuvant treatment. BRIEF DESCRIPTION OF THE DRAWINGS

[0007] Figure 1 includes two line graphs, illustrating Kaplan-Meier survival curves for TP53 mutational status in mCRC patients showing improved OS (Figure 1A) and PFS (Figure 1B) with BOLD-100 treatment.

[0008] Figure 2 includes two line graphs, illustrating Kaplan-Meier survival curves for TP53 mutational status in mCRC patients with gain-of-function (GOF) and non-GOF mutations, showing improved OS (Figure 1A) and PFS (Figure 1B) with BOLD-100 treatment particularly in patients with GOF mutations.

[0009] Figure 3 includes two line graphs, illustrating Kaplan-Meier survival curves for TP53 mutational status in mCRC patients with gain-of-function (GOF) mutations by sidedness, showing improved OS (Figure 3A) and PFS (Figure 3B) with BOLD-100 treatment mediating improved responses in these patients independent of tumour sidedness.

[0010] Figure 4 sets out the canonical 393 amino acid long sequence of the p53 protein, isoform P04637-1, SEQ ID NO:2.DETAILED DESCRIPTION

[0011] As disclosed herein, BOLD-100 is shown to have a significant impact on the clinical outcomes of patients undergoing chemotherapy for cancers having mutations in the TP53 gene. The NCBI Reference Sequence for the TP53 gene is set out in NCBI Reference Sequence: NG_017013.2:5001-24149 Homo sapiens tumor protein p53 (TP53), RefSeqGene (LRG_321) on chromosome 17 (SEQ ID NO:1). Genomic TP53 mutations relevant to treatment with BOLD- 100 may include any substitutions, deletions or insertions in the TP53 gene sequence, including mutations that affect the sequence of the p53 protein encoded by the gene.

[0012] The canonical protein sequence of p53 is identified in the UniProt database by reference P04637-1, having the 393 amino acid long sequence set out in Figure 4 (SEQ ID NO:2), with alternative isoforms identified by sequences P04637-2 though P04637-9. Analysis presented in the Examples herein identifies relevant TP53 mutations as for example including mutations at positions encoding amino acids in the p53 protein, including positions: 175, 245, 248, 249, 273, or 282. These may for example include the substation of the canonical amino acid at this positions for any other amino acid (“X”), i.e.: R175X, G245X, R248X, R249X, R273X, R273X, or R282X. Recognized variants at these positions for example include R175A, R175C, R175G, R175H, R175L R175P, G245A, G245C, G245D, G245F, G245R, G245S, G245V, R248E, R248G, R248L, R248P, R248Q, R248W, R249G, R249K, R249M, R249S, R249T, R249W, R273C, R273H, R273L, R273P, R273S, R273W, R273Y, R282D, R282G, R282H, R282Q and R282W. Alternative mutations include frameshift mutations (fs), insertions (ins), deletions (del) and truncations (*), including such mutations that affect positions R175, G245, R248, R248, R249, R273, R273, or R282. Gain of function (GOF) mutations, or mutations that are identified as conferring at least some GOF, identified in The Clinical Knowledgebase (CKB) managed by The Jackson Laboratory include: S121A, S121C, S121F, C124S, V157S, A161T, R175H, L194R, T211A, F212Y, H214R, G245A, G245C, G245D, G245S, R248G, R249W, R273H, C277S, C277Y, R280K, D281G, R282Q, R290G, K291E, K291Q, K291T, K291I, K292T, and G360V. Relevant mutations may accordingly include other variants at these positions: S121X, C124X, V157X, A161X, R175X, L194X, T211X, F212X, H214X, G245X, R248X, R249X, R273X, C277X, R280X, D281X, R282X, R290X, K291X, K292X, and G360X. In select embodiments, relevant GOF mutations herein may for example include: R175H, G245S, R248Q, R248W, R249S, R273H, R273L, or R282W.

[0013] In various aspects, treatments are provided that involve the use of BOLD-100 alone or in combination with other chemotherapeutic agents to treat TP53MT cancers. In an exemplified embodiment, BOLD-100 was used in combination with FOLFOX in treatment of metastatic colorectal cancers (mCRC), as disclosed in detail in the Examples herein. FOLFOX treatment comprises use of the drugs folinic acid (leucovorin, FOL), fluorouracil (5-FU, F), and oxaliplatin (Eloxatin, OX). In alternative embodiments, BOLD-100 may be used with a variety of implementations of FOLFOX therapy. For example, a FOLFOX4 dosing schedule may be adopted as follows, in cycles that may be repeated every two weeks: • Day 1: Oxaliplatin 85 mg / m2intravenous (IV) infusion in 250-500 mL D5W and leucovorin 200 mg / m2IV infusion in D5W administered concurrently over 120 minutes in separate bags using a Y-line, followed by fluorouracil (5-FU) 400 mg / m2IV bolus given over 2–4 minutes, followed by 5-FU 600 mg / m2IV infusion in 500 mL D5W as a 22-hour continuous infusion. • Day 2: Leucovorin 200 mg / m2IV infusion over 120 minutes, followed by 5-FU 400 mg / m2IV bolus given over 2–4 minutes, followed by 5-FU 600 mg / m2IV infusion in 500 mL D5W as a 22-hour continuous infusion.

[0014] An alternative FOLFOX6 dosing schedule may be adopted as follows, in cycles that may be repeated every two weeks: • Day 1–2: Oxaliplatin 100 mg / m2IV infusion, given as a 120 minutes IV infusion in 500 mL D5W, concurrent with leucovorin 400 mg / m2(or levoleucovorin 200 mg / m2) IV infusion, followed by 5-FU 400 mg / m2 IV bolus, followed by 46-hour 5- FU infusion (2400 mg / m2for first two cycles, and may be increased to 3000 mg / m2if tolerated by patient (no toxicity > grade 1 during the first two cycles).

[0015] Patients may for example receive BOLD-100 treatment with FOLFOX on day 1 or day 2 of each 14-day cycle. BOLD-100 may for example be administered at a dosage of 320, 420, 500, or 625 mg / m2, for example in a dosing regimen of BOLD-100 in combination with FOLFOX chemotherapy, as follows: • 320 mg / m2IV over 30-60 minutes, or 30-90 minutes, or 60-90 minutes, every two weeks; • 420 mg / m2IV over 30-60 minutes, or 30-90 minutes, or 60-90 minutes, every two weeks;• 500 mg / m2IV over 30-60 minutes, or 30-90 minutes, or 60-90 minutes, every two weeks; or • 625 mg / m2IV over 30-60 minutes, or 30-90 minutes, or 60-90 minutes, every two weeks.

[0016] A very wide range of TP53MT cancers are amenable to treatment with BOLD-100 alone or in combination. TP53 is generally understood to be the most mutated cancer gene collectively over all cancers. Querying the cBioPortal database of 76363 patients (all cancer types and stages, using a curated set of non-redundant studies in 2024), 33% had a TP53 mutation (as is reflected in the literature, see Chen, et al. Mutant p53 in cancer: from molecular mechanism to therapeutic modulation. Cell Death Dis 13, 974 (2022)). BOLD-100 may for example be combined with a variety of therapies targeting cancers having TP53 mutations (see Nishikawa and Iwakuma, Drugs Targeting p53 Mutations with FDA Approval and in Clinical Trials. Cancers (Basel).2023 Jan 9;15(2):429). Relevant cancers include: advanced solid tumors and malignancies (bladder, gastric, NSCLC, urothelial); myelodysplastic syndromes (MDS); oligoblastic acute myeloid leukemia (AML), such as AML / MDS in post-hematopoietic stem-cell transplant (HCT) maintenance therapy; platinum-sensitive recurrent high grade serous ovarian cancer (HGSOC); platinum-resistant recurrent HGSOC; myeloid malignancies; oral cancers; refractory solid tumors; recurrent and metastatic ovarian and endometrial cancer; high- grade platinum-resistant ovarian cancer; solid tumor and re-lapsed AML; refractory and resistant ovarian cancer; platinum-sensitive ovarian tumors; platinum-resistant ovarian cancer; metastatic colorectal cancer with RAS; recurrent uterine serous carcinoma; relapsed small cell lung cancer (SCLC) with cyclin-dependent kinase inhibitor 2A (CDKN2A) mutations; untreated stage IV non-small cell lung cancer (NSCLC); NSCLC; triple negative breast cancer.

[0017] Specific cancers in which the cBioPortal database discloses that at least approximately 10% of cancers include TP53 mutations, and accordingly amenable to BOLD-100 treatment alone or in combination, include: large cell lung carcinoma; diffuse astrocytoma; high- grade serous ovarian cancer; non small cell lung cancer other; ovarian epithelial tumor; poorly differentiated non-small cell lung cancer; uterine serous carcinoma / uterine papillary serous carcinoma; esophageal squamous cell carcinoma; ovarian carcinosarcoma / malignant mixed mesodermal tumor; uterine carcinosarcoma / uterine malignant mixed mullerian tumor; adenosquamous carcinoma of the pancreas; lung squamous cell carcinoma; small cell lung cancer; pancreatic cancer other; esophageal carcinoma other; ovarian adenocarcinoma; gastriccarcinoma other; gallbladder carcinoma other; esophageal adenocarcinoma; cutaneous squamous cell carcinoma; mucinous ovarian cancer; gliosarcoma; adenocarcinoma of the gastroesophageal junction; gallbladder adenocarcinoma; larynx squamous cell carcinoma; biliary tract; uterine mixed endometrial carcinoma; lung adenosquamous carcinoma; rectal adenocarcinoma; undifferentiated sarcoma; serous ovarian cancer; large cell neuroendocrine carcinoma; pancreatic adenocarcinoma; astrocytoma; anaplastic oligoastrocytoma; oral cavity squamous cell carcinoma; metaplastic breast cancer; anaplastic astrocytoma; basal cell carcinoma; salivary duct carcinoma; high-grade neuroendocrine carcinoma of the colon and rectum; pancreatobiliary ampullary carcinoma; colorectal adenocarcinoma; burkitt lymphoma; tubular stomach adenocarcinoma; extrahepatic cholangiocarcinoma; undifferentiated pleomorphic sarcoma malignant fibrous histiocytoma; signet ring cell carcinoma of the stomach; head and neck squamous cell carcinoma; skin cancer, non-melanoma; liver hepatocellular carcinoma plus intrahepatic cholangiocarcinoma; gastric adenocarcinoma; colon adenocarcinoma; urothelial carcinoma other; ampullary carcinoma; oligoastrocytoma; small bowel cancer; leiomyosarcoma; pleomorphic liposarcoma; liver hepatocellular carcinoma; ovarian serous carcinoma; sarcomatoid carcinoma of the lung; intestinal type stomach adenocarcinoma; uterine leiomyosarcoma; myxoid fibrosarcoma; uterine clear cell carcinoma; breast invasive carcinoma; breast invasive carcinoma, nos; esophagogastric adenocarcinoma; small bowel adenocarcinoma; cancer of unknown primary; undifferentiated pleomorphic sarcoma / malignant fibrous histiocytoma / high-grade spindle cell sarcoma; breast carcinoma other; pleomorphic rhabdomyosarcoma; uterine perivascular epithelioid cell tumor; penile squamous cell carcinoma; mucinous stomach adenocarcinoma; neuroendocrine carcinoma, nos; adenocarcinoma, nos; poorly differentiated carcinoma, nos; stomach adenocarcinoma; anaplastic thyroid cancer; lung adenocarcinoma; perihilar cholangiocarcinoma; appendiceal adenocarcinoma; head and neck squamous cell carcinomaãš; papillary stomach adenocarcinoma; bladder urothelial carcinoma; non-small cell lung cancer; high-grade glioma, nos; bladder squamous cell carcinoma; prostate neuroendocrine carcinoma; breast invasive ductal carcinoma; glioblastoma; mucinous cystic neoplasm; hepatocellular carcinoma plus intrahepatic cholangiocarcinoma; myxofibrosarcoma; gallbladder cancer; breast invasive cancer, nos; lung neuroendocrine tumor; invasive breast carcinoma; non-hodgkin lymphoma; burkitt lymphoma; perivascular epithelioid cell tumor; diffuse type stomach adenocarcinoma; malignant phyllodes tumor of the breast; squamous cell carcinoma of the vulva / vagina;intrahepatic cholangiocarcinoma; chondroblastic osteosarcoma; angiosarcoma; cholangiocarcinoma; mucinous adenocarcinoma of the appendix; mucinous adenocarcinoma of the colon and rectum; osteosarcoma; sarcoma, nos; chondrosarcoma; hepatocellular carcinoma; mixed cancer types; mature b-cell neoplasms; merkel cell carcinoma; endometrial adenocarcinoma; glioblastoma multiforme; mucinous carcinoma; desmoplastic melanoma; chromophobe renal cell carcinoma; pediatric high grade gliomas; mucosal melanoma of the vulva / vagina; melanoma of unknown primary; embryonal rhabdomyosarcoma; intraductal papillary mucinous neoplasm; choriocarcinoma; prostate adenocarcinoma; solitary fibrous tumor / hemangiopericytoma; yolk sac tumor; infiltrating ductal carcinoma; diffuse large b-cell lymphoma; hepatocellular adenoma; endometrioid ovarian cancer; breast invasive carcinoma (nos); therapy-related myeloid neoplasms; endometrial carcinoma; aml with myelodysplasia- related changes; thymic carcinoma; aml with myelodysplasia-related changes; rhabdomyosarcoma; therapy-related myeloid neoplasms; unclassified kidney renal cell carcinoma; myeloid neoplasm; mantle cell lymphoma; upper tract urothelial carcinoma; adrenocortical carcinoma; uterine endometrioid carcinoma; head and neck carcinoma other; pleural mesothelioma, biphasic type; fibrosarcoma; giant cell tumor of bone; diffuse glioma; malignant peripheral nerve sheath tumor; breast mixed ductal and lobular carcinoma; activated b-cell type; oligodendroglioma; spindle cell sarcoma; prostate; breast; liposarcoma; germinal center b-cell type; peripheral t-cell lymphoma, nos; hurthle cell thyroid cancer; myxoid / round-cell liposarcoma; castration-resistant prostate cancer; melanoma; cutaneous melanoma; poorly differentiated thyroid cancer; oropharynx squamous cell carcinoma; endometrial stromal sarcoma; sezary syndrome; pleural mesothelioma, epithelioid type; malignant tumor; ewing sarcoma; adenoid cystic breast cancer; breast invasive lobular carcinoma; soft tissue sarcoma other; small cell carcinoma of the ovary; pancreatic neuroendocrine tumor; ossifying fibromyxoid tumor; head and neck mucosal melanoma; follicular lymphoma; anorectal mucosal melanoma; mucinous liposarcoma; anal squamous cell carcinoma; acute myeloid leukemia; myelodysplastic syndromes; diffuse large b-cell lymphoma, nos; nasopharyngeal carcinoma; fibroblastic myofibroblastic tumor; acinar cell carcinoma of the pancreas; mixed germ cell tumor; and, kidney renal cell carcinoma.

[0018] The cBioPortal database identifies cancers having GOF TP53 mutations, and accordingly amenable to BOLD-100 treatment alone or in combination, including adenocarcinoma of the gastroesophageal junction; gallbladder carcinoma other; undifferentiatedsarcoma; uterine serous carcinoma / uterine papillary serous carcinoma; mucinous ovarian cancer; rectal adenocarcinoma; esophageal adenocarcinoma; burkitt lymphoma; gliosarcoma; uterine carcinosarcoma / uterine malignant mixed mullerian tumor; mucinous cystic neoplasm; colorectal adenocarcinoma; gastric carcinoma other; pancreatic cancer other; ovarian adenocarcinoma; myxoid fibrosarcoma; small bowel cancer; non-hodgkin lymphoma; colon adenocarcinoma; liver hepatocellular carcinoma plus intrahepatic cholangiocarcinoma; high- grade neuroendocrine carcinoma of the colon and rectum; pancreatobiliary ampullary carcinoma; tubular stomach adenocarcinoma; astrocytoma; diffuse astrocytoma; high-grade serous ovarian cancer; ovarian carcinosarcoma / malignant mixed mesodermal tumor; cutaneous squamous cell carcinoma; uterine clear cell carcinoma; uterine mixed endometrial carcinoma; signet ring cell carcinoma of the stomach; mucinous adenocarcinoma of the appendix; anaplastic astrocytoma; ovarian serous carcinoma; large cell lung carcinoma; salivary duct carcinoma; high-grade glioma, nos; small bowel adenocarcinoma; pancreatic adenocarcinoma; metaplastic breast cancer; intraductal papillary mucinous neoplasm; basal cell carcinoma; pleomorphic liposarcoma; anaplastic oligoastrocytoma; head and neck squamous cell carcinoma; therapy-related myeloid neoplasms; liver hepatocellular carcinoma; biliary tract; esophagogastric adenocarcinoma; ampullary carcinoma; esophageal squamous cell carcinoma; gastric adenocarcinoma; oral cavity squamous cell carcinoma; bladder squamous cell carcinoma; hepatocellular carcinoma plus intrahepatic cholangiocarcinoma; extrahepatic cholangiocarcinoma; head and neck squamous cell carcinoma; burkitt lymphoma; stomach adenocarcinoma; oligoastrocytoma; squamous cell carcinoma of the vulva / vagina; neuroendocrine carcinoma, nos; mature b-cell neoplasms.

[0019] BOLD-100 may be used in combination with other agents for treating TP53MT cancers, including: platinum-based chemotherapeutics (e.g. cisplatin, carboplatin, oxaliplatin); antimetabolites (e.g. methotrexate, 5-FU, aminopterin, cytarabine, thioguanine, and mercaptopurine); anthracyclines (e.g daunorubicin, doxorubicin, epirubicin, idarubicin, mitoxantrone or valrubicin); EGFR targeted therapies; PARPi targeted therapies; VEGF inhibitors, VEGFR inhibitors, bevacizumab, and immunotherapies.

[0020] Specific cancers in which the database discloses that at least approximately 10% of cancer include TP53 mutations include: large cell lung carcinoma; diffuse astrocytoma; high- grade serous ovarian cancer; non-small cell lung cancer other; ovarian epithelial tumor; poorly differentiated non-small cell lung cancer; uterine serous carcinoma / uterine papillary serouscarcinoma; esophageal squamous cell carcinoma; ovarian carcinosarcoma / malignant mixed mesodermal tumor; uterine carcinosarcoma / uterine malignant mixed mullerian tumor; adenosquamous carcinoma of the pancreas; lung squamous cell carcinoma; small cell lung cancer; pancreatic cancer other; esophageal carcinoma other; ovarian adenocarcinoma; gastric carcinoma other; gallbladder carcinoma other; esophageal adenocarcinoma; cutaneous squamous cell carcinoma; mucinous ovarian cancer; gliosarcoma; adenocarcinoma of the gastroesophageal junction; gallbladder adenocarcinoma; larynx squamous cell carcinoma; biliary tract; uterine mixed endometrial carcinoma; lung adenosquamous carcinoma; rectal adenocarcinoma; undifferentiated sarcoma; serous ovarian cancer; large cell neuroendocrine carcinoma; pancreatic adenocarcinoma; astrocytoma; anaplastic oligoastrocytoma; oral cavity squamous cell carcinoma; metaplastic breast cancer; anaplastic astrocytoma; basal cell carcinoma; salivary duct carcinoma; high-grade neuroendocrine carcinoma of the colon and rectum; pancreatobiliary ampullary carcinoma; colorectal adenocarcinoma; Burkitt lymphoma; tubular stomach adenocarcinoma; extrahepatic cholangiocarcinoma; undifferentiated pleomorphic sarcoma malignant fibrous histiocytoma; signet ring cell carcinoma of the stomach; head and neck squamous cell carcinoma; skin cancer, non-melanoma; liver hepatocellular carcinoma plus intrahepatic cholangiocarcinoma; gastric adenocarcinoma; colon adenocarcinoma; urothelial carcinoma other; ampullary carcinoma; oligoastrocytoma; small bowel cancer; leiomyosarcoma; pleomorphic liposarcoma; liver hepatocellular carcinoma; ovarian serous carcinoma; sarcomatoid carcinoma of the lung; intestinal type stomach adenocarcinoma; uterine leiomyosarcoma; myxoid fibrosarcoma; uterine clear cell carcinoma; breast invasive carcinoma; breast invasive carcinoma, nos; esophagogastric adenocarcinoma; small bowel adenocarcinoma; cancer of unknown primary; undifferentiated pleomorphic sarcoma / malignant fibrous histiocytoma / high-grade spindle cell sarcoma; breast carcinoma other; pleomorphic rhabdomyosarcoma; uterine perivascular epithelioid cell tumor; penile squamous cell carcinoma; mucinous stomach adenocarcinoma; neuroendocrine carcinoma, nos; adenocarcinoma, nos; poorly differentiated carcinoma, nos; stomach adenocarcinoma; anaplastic thyroid cancer; lung adenocarcinoma; perihilar cholangiocarcinoma; appendiceal adenocarcinoma; head and neck squamous cell carcinomaãš; papillary stomach adenocarcinoma; bladder urothelial carcinoma; non-small cell lung cancer; high-grade glioma, nos; bladder squamous cell carcinoma; prostate neuroendocrine carcinoma; breast invasive ductal carcinoma; glioblastoma; mucinous cystic neoplasm; hepatocellular carcinoma plusintrahepatic cholangiocarcinoma; myxofibrosarcoma; gallbladder cancer; breast invasive cancer, nos; lung neuroendocrine tumor; invasive breast carcinoma; non-hodgkin lymphoma; burkitt lymphoma; perivascular epithelioid cell tumor; diffuse type stomach adenocarcinoma; malignant phyllodes tumor of the breast; squamous cell carcinoma of the vulva / vagina; intrahepatic cholangiocarcinoma; chondroblastic osteosarcoma; angiosarcoma; cholangiocarcinoma; mucinous adenocarcinoma of the appendix; mucinous adenocarcinoma of the colon and rectum; osteosarcoma; sarcoma, nos; chondrosarcoma; hepatocellular carcinoma; mixed cancer types; mature b-cell neoplasms; Merkel cell carcinoma; endometrial adenocarcinoma; glioblastoma multiforme; mucinous carcinoma; desmoplastic melanoma; chromophobe renal cell carcinoma; pediatric high grade gliomas; mucosal melanoma of the vulva / vagina; melanoma of unknown primary; embryonal rhabdomyosarcoma; intraductal papillary mucinous neoplasm; choriocarcinoma; prostate adenocarcinoma; solitary fibrous tumor / hemangiopericytoma; yolk sac tumor; infiltrating ductal carcinoma; diffuse large b-cell lymphoma; hepatocellular adenoma; endometrioid ovarian cancer; breast invasive carcinoma (nos); therapy-related myeloid neoplasms; endometrial carcinoma; aml with myelodysplasia- related changes; thymic carcinoma; aml with myelodysplasia-related changes; rhabdomyosarcoma; therapy-related myeloid neoplasms; unclassified kidney renal cell carcinoma; myeloid neoplasm; mantle cell lymphoma; upper tract urothelial carcinoma; adrenocortical carcinoma; uterine endometrioid carcinoma; head and neck carcinoma other; pleural mesothelioma, biphasic type; fibrosarcoma; giant cell tumor of bone; diffuse glioma; malignant peripheral nerve sheath tumor; breast mixed ductal and lobular carcinoma; activated b-cell type; oligodendroglioma; spindle cell sarcoma; prostate; breast; liposarcoma; germinal center b-cell type; peripheral t-cell lymphoma, nos; hurthle cell thyroid cancer; myxoid / round-cell liposarcoma; castration-resistant prostate cancer; melanoma; cutaneous melanoma; poorly differentiated thyroid cancer; oropharynx squamous cell carcinoma; endometrial stromal sarcoma; sezary syndrome; pleural mesothelioma, epithelioid type; malignant tumor; ewing sarcoma; adenoid cystic breast cancer; breast invasive lobular carcinoma; soft tissue sarcoma other; small cell carcinoma of the ovary; pancreatic neuroendocrine tumor; ossifying fibromyxoid tumor; head and neck mucosal melanoma; follicular lymphoma; anorectal mucosal melanoma; mucinous liposarcoma; anal squamous cell carcinoma; acute myeloid leukemia; myelodysplastic syndromes; diffuse large b-cell lymphoma, nos; nasopharyngeal carcinoma;fibroblastic myofibroblastic tumor; acinar cell carcinoma of the pancreas; mixed germ cell tumor; and, kidney renal cell carcinoma.

[0021] Treatments of TP53MT cancers involving the use of BOLD-100 in combination with an antimetabolite may for example include treatment with: 5-FU for the treatment of breast cancer, colorectal cancer, stomach cancer, pancreatic cancer, skin cancer; azacitidine in the treatment of acute myeloid leukemia, myelodysplastic syndromes; capecitabine in the treatment of breast cancer, colorectal cancer, stomach cancer, esophageal cancer, pancreatic cancer; cladribine in the treatment of hairy cell leukemia; clofarabine in the treatment of acute lymphoblastic leukemia; cytarabine in the treatment of acute non-lymphocytic leukemia, meningeal leukemia, acute lymphoblastic leukemia (ALL), chronic myelogenous leukemia (CML); decitabine in the treatment of myelodysplastic syndromes; floxuridine in the treatment of stomach cancer; fludarabine phosphate in the treatment of chronic lymphocytic leukemia; gemcitabine in the treatment of breast cancer, non-small cell lung cancer, ovarian cancer, pancreatic cancer; hydroxyurea in the treatment of chronic myelogenous leukemia, squamous cell carcinoma of the head and neck; methotrexate in the treatment of acute lymphoblastic leukemia, breast cancer, head and neck cancer, lung cancer, mycosis fungoides, non-Hodgkin lymphoma, osteosarcoma; nelarabine in the treatment of T-cell acute lymphoblastic leukemia, T-cell lymphoblastic lymphoma; pemetrexed in the treatment of pleural mesothelioma, non- squamous non-small cell lung cancer; pentostatin in the treatment of hairy cell leukemia; pralatrexate in the treatment of peripheral T-cell lymphoma; thioguanine in the treatment of acute myeloid leukemia; trifluridine / tipiracil in the treatment of colorectal cancer, or stomach cancer.

[0022] Treatments of TP53MT cancers involving the use of BOLD-100 in combination with an anthracycline may for example include treatment with daunorubicin, doxorubicin, epirubicin, idarubicin, mitoxantrone or valrubicin. Cancers amenable to such treatment include acute lymphocytic leukemia, acute myelogenous leukemia, Hodgkin’s lymphoma, non-Hodgkin’s lymphoma, bladder cancer, rapidly recurrent (ta or ti) or in situ transitional cell carcinoma (intravesical therapy), neoadjuvant treatment, metastatic transitional cell bladder cancer, breast cancer, adjuvant therapy following axillary lymph node resection in primary breast cancer, neoadjuvant therapy, metastatic breast cancer, other metastatic cancers, ovarian cancer, osteogenic sarcoma, ewing sarcoma, soft tissue sarcoma, thyroid cancer, neuroblastoma, wilm's tumor, and small cell lung cancer.

[0023] Treatments of TP53MT cancers involving the use of BOLD-100 in combination with an EGFR targeted therapies may for example include treatment of NSCLC with afatinib, gefitnib, amivantamab-vmjw, osimertinib, erlotinib, or dacomitinib.

[0024] Treatments of TP53MT cancers involving the use of BOLD-100 in combination with a platinum chemotherapeutic may for example include treatment with cisplatin, carboplatin, and / or oxaliplatin of: advanced ovarian cancer, testicular cancer, and bladder carcinoma; breast cancer; cervical and endometrial carcinoma; gestational trophoblastic neoplasia; hormone- sensitive and Her2neu-positive tumors; triple-negative breast cancer; esophageal cancer ; gastric cancer; hepatobiliary cancer; advanced cervical cancer; lung cancer, both small and non-small cells; metastatic, advanced, and refractory cancers; Hodgkin lymphoma; non-Hodgkin lymphoma; penile cancer; thymoma; head and neck cancers; osteosarcoma; multiple myeloma; and mesothelioma.

[0025] Treatments of TP53MT cancers involving the use of BOLD-100 in combination with a poly (ADP-ribose) polymerase (PARP) inhibitor may for example include treatment with olaparib, rucaparib, or niraparib, for: pancreatic cancer, prostate cancer, ovarian cancer, fallopian tube cancer, peritoneal cancer, lung cancer, pancreatic cancer, head and neck cancer, glioblastoma multiforme, prostate cancer, stomach cancer, oesophageal cancer, womb and cervical cancer, kidney and bladder cancer.

[0026] Treatments of TP53MT cancers involving the use of BOLD-100 in combination with an epidermal growth factor receptor (EGFR) inhibitor may for example include treatment with tyrosine kinase inhibitors (e.g., afatinib, dacomitinib, erlotinib, gefitinib, osimertinib) or monoclonal anti-EGFR antibodies (e.g., cetuximab, panitumumab, nimotuzumab, necitumumab), to treat cancers characterized by EGFR upregulation, including non-small-cell lung cancer, pancreatic cancer, breast cancer, and colon cancer.

[0027] Treatments of TP53MT cancers involving the use of BOLD-100 in combination with a vascular endothelial growth factor (VEGF) or vascular endothelial growth factor receptor (VEGFR) inhibitor may for example include treatment with sunitinib, pazopanib, foretinib, sorafenib, tivozanib, anlotinib, axitinib, golvatinib, fresolimumab, trastuzumab, ramucirumab, tanibirumab, sym004, bevacizumab, aflibercept, or sevacizumab. These treatments may for example include the treatment of advanced renal carcinoma, meninginoma, pancreatic cancer, soft tissue sarcoma, head / neck cancers, gastric cancers, solid tumours, colorectal cancer, stroma cancer, breast cancer, liver cancer, hepatocellular carcinoma,malignant melanoma, renal cancer, breast cancer, thyroid carcinoma, advanced gastric cancer, recurrent glioblastoma, or metastatic colorectal cancer.

[0028] Treatments of TP53MT cancers involving the use of BOLD-100 in combination with an immunotherapeutic, may for example involve treatment with an immune checkpoint inhibitor, a T-cell transfer therapy, a monoclonal antibody (e.g. alemtuzumab, atezolizumab, Avelumab, ipilimumab, elotuzumab, ofatumumab, nivolumab, pembrolizumab, rituximab or durvalumab), a therapeutic cancer vaccine, or an immune system modulator.

[0029] Additional embodiments of the present invention provide methods for preparing drug products containing the sodium salt of trans-[tetrachlorobis(1H-indazole)ruthenate (III)] (i.e. BOLD-100).

[0030] One aspect of the current invention provides a method for preparing a sterile, lyophilized drug product containing sodium trans-[tetrachlorobis(1H-indazole)ruthenate (III)]. This formulation would be suitable for administration to a patient. The formulation is comprised of sodium trans-[tetrachlorobis(1H-indazole)ruthenate (III)], a pH buffer, and a cryoprotective agent. The general method for providing said formulation comprises the steps of preparing aqueous buffer solution, preparing aqueous cryoprotectant solution, dissolution of sodium trans- [tetrachlorobis(1H-indazole)ruthenate (III)] in the buffer solution, addition of the cryoprotectant solution, sterile filtration (e.g. aseptic filtration), filling of vials under sterile conditions, and lyophilization under sterile conditions. Suitable buffers include, but are not limited to: citrate, TRIS, acetate, EDTA, HEPES, tricine, and imidazole. The use of a phosphate buffer is possible but is not preferred. A preferred aspect of the present invention is the use of a citric acid / sodium citrate buffer. Suitable cryoprotective agents include, but are not limited to: sugars, monosaccharides, disaccharides, polyalcohols, mannitol, sorbitol, sucrose, trehalose, dextran, and dextrose. A preferred aspect of the present invention is the use of mannitol as the cyroprotecive agent.

[0031] Sodium trans-[tetrachlorobis(1H-indazole)ruthenate (III)] can degrade in water. Limiting this degradation reaction may be advantageous to obtaining the highest purity product. Cooling the sodium trans-[tetrachlorobis(1H-indazole)ruthenate (III)] solution during the formulation process may reduce the amount of degraded product present in the lyophilized product. For example, the sodium trans-[tetrachlorobis(1H-indazole)ruthenate (III)]solution may be cooled to 4 °C during the formulation process, or cooled to 2-8 °C during the formulation process, or cooled to 2-15 °C during the formulation process.

[0032] One embodiment of the present invention provides a composition comprising sodium trans-[tetrachlorobis(1H-indazole)ruthenate (III)], a suitable buffer, and mannitol. In some embodiments, a suitable buffer comprises a citrate buffer. For instance, in some embodiments, a citrate buffer comprises sodium citrate and citric acid. One embodiment of the present invention provides a composition comprising sodium trans-[tetrachlorobis(1H- indazole)ruthenate (III)], sodium citrate, citric acid, and mannitol. One embodiment of the present invention provides a composition comprising sodium trans-[tetrachlorobis(1H- indazole)ruthenate (III)], sodium citrate, citric acid, mannitol, and mer,trans- [RuIIICl3(Hind)2(H2O)]. One embodiment of the present invention provides a composition comprising sodium trans-[tetrachlorobis(1H-indazole)ruthenate (III)], sodium citrate, citric acid, mannitol, mer,trans-[RuIIICl3(Hind)2(H2O)], and a cesium salt. One embodiment of the present invention provides a composition comprising sodium trans-[tetrachlorobis(1H-indazole)ruthenate (III)], sodium citrate, citric acid, and mannitol, wherein the sodium trans-[tetrachlorobis(1H- indazole)ruthenate (III)] is amorphous. One embodiment of the present invention provides a composition comprising sodium trans-[tetrachlorobis(1H-indazole)ruthenate (III)], sodium citrate, citric acid, mannitol, and mer,trans-[RuIIICl3(Hind)2(H2O)], wherein the sodium trans- [tetrachlorobis(1H-indazole)ruthenate (III)] is amorphous. One embodiment of the present invention provides a composition comprising sodium trans-[tetrachlorobis(1H-indazole)ruthenate (III)], sodium citrate, citric acid, mannitol, mer,trans-[RuIIICl3(Hind)2(H2O)], and a cesium salt, wherein the sodium trans-[tetrachlorobis(1H-indazole)ruthenate (III)] is amorphous.

[0033] One embodiment of the present invention provides a composition comprising sodium trans-[tetrachlorobis(1H-indazole)ruthenate (III)], sodium citrate, citric acid, mannitol, mer,trans-[RuIIICl3(Hind)2(H2O)], and a cesium salt; wherein: mer,trans-[RuIIICl3(Hind)2(H2O)] is between about 0.01 and about 0.4 weight percent of the composition, and cesium is between about 0.00001 and about 0.01 weight percent of the composition.

[0034] One embodiment of the present invention provides a composition comprising sodium trans-[tetrachlorobis(1H-indazole)ruthenate (III)], sodium citrate, citric acid, mannitol, mer,trans-[RuIIICl3(Hind)2(H2O)], and a cesium salt; wherein:mer,trans-[RuIIICl3(Hind)2(H2O)] is between about 0.01 and about 0.4 weight percent of the composition, and cesium is between about 0.00001 and about 0.01 weight percent of the composition.

[0035] One embodiment of the present invention provides a composition comprising sodium trans-[tetrachlorobis(1H-indazole)ruthenate (III)], sodium citrate, citric acid, mannitol, mer,trans-[RuIIICl3(Hind)2(H2O)], and a cesium salt; wherein: mer,trans-[RuIIICl3(Hind)2(H2O)] is between about 0.01 and about 0.2 weight percent of the composition, and cesium is between about 0.00001 and about 0.01 weight percent of the composition.

[0036] One embodiment of the present invention provides a composition comprising sodium trans-[tetrachlorobis(1H-indazole)ruthenate (III)], mer,trans-[RuIIICl3(Hind)2(H2O)], and a cesium salt; wherein: mer,trans-[RuIIICl3(Hind)2(H2O)] is between about 0.01 and about 0.40 weight percent of the composition, and cesium is between about 0.00001 and about 0.01 weight percent of the composition.

[0037] One embodiment of the present invention provides a composition comprising sodium trans-[tetrachlorobis(1H-indazole)ruthenate (III)], mer,trans-[RuIIICl3(Hind)2(H2O)], and a cesium salt; wherein: the composition is a lyophilized powder, mer,trans-[RuIIICl3(Hind)2(H2O)] is between about 0.01 and about 0.40 weight percent of the composition, and cesium is between about 0.00001 and about 0.01 weight percent of the composition.

[0038] One embodiment of the present invention provides a composition comprising sodium trans-[tetrachlorobis(1H-indazole)ruthenate (III)], sodium citrate, citric acid, mannitol, mer,trans-[RuIIICl3(Hind)2(H2O)], and a cesium salt; wherein: the composition is a lyophilized powder, mer,trans-[RuIIICl3(Hind)2(H2O)] is between about 0.01 and about 0.3 weight percent of the composition,and cesium is between about 0.00001 and about 0.1 weight percent of the composition.

[0039] One embodiment of the present invention provides a composition comprising sodium trans-[tetrachlorobis(1H-indazole)ruthenate (III)], sodium citrate, citric acid, mannitol, mer,trans-[RuIIICl3(Hind)2(H2O)], and a cesium salt; wherein: mer,trans-[RuIIICl3(Hind)2(H2O)] is between about 0.01 and about 0.3 weight percent of the composition, and cesium is between about 0.00001 and about 0.1 weight percent of the composition.

[0040] One embodiment of the present invention provides a composition comprising sodium trans-[tetrachlorobis(1H-indazole)ruthenate (III)], sodium citrate, citric acid, mannitol, mer,trans-[RuIIICl3(Hind)2(H2O)], and a cesium salt; wherein: the composition is a lyophilized powder, sodium trans-[tetrachlorobis(1H-indazole)ruthenate (III)] is about 11.5 to about 14.0 weight percent of the compositon, citric acid is about 43.9 to about 53.7 weight percent of the composition, sodium citrate is about 25.7 to about 23.1 weight percent of the composition, mannitol is about 11.5 to about 14.0 weight percent of the composition, mer,trans-[RuIIICl3(Hind)2(H2O)] is about 0.01 and about 0.3 weight percent of the composition, and cesium is between about 0.00001 and about 0.1 weight percent of the composition.

[0041] One embodiment of the present invention provides a composition comprising sodium trans-[tetrachlorobis(1H-indazole)ruthenate (III)], sodium citrate, citric acid, mannitol, mer,trans-[RuIIICl3(Hind)2(H2O)], and a cesium salt; wherein: the composition is a lyophilized powder, sodium trans-[tetrachlorobis(1H-indazole)ruthenate (III)] is about 10.2 to about 15.3 weight percent of the composition, citric acid is about 39.0 to about 58.5 weight percent of the composition, sodium citrate is about 20.5 to about 30.8 weight percent of the compositon, mannitol is about 10.2 to about 15.3 weight percent of the composition,mer,trans-[RuIIICl3(Hind)2(H2O)] is about 0.01 and about 0.3 weight percent of the composition, and cesium is between about 0.00001 and about 0.1 weight percent of the composition.

[0042] One embodiment of the present invention provides a composition comprising sodium trans-[tetrachlorobis(1H-indazole)ruthenate (III)], sodium citrate, citric acid, mannitol, mer,trans-[RuIIICl3(Hind)2(H2O)], and a cesium salt; wherein: the composition is a lyophilized powder, sodium trans-[tetrachlorobis(1H-indazole)ruthenate (III)] is about 10.2 to about 15.3 weight percent of the composition, mer,trans-[RuIIICl3(Hind)2(H2O)] is about 0.01 and about 0.3 weight percent composition, and cesium is between about 0.00001 and about 0.1 weight percent of the composition.

[0043] One embodiment of the present invention provides a composition comprising sodium trans-[tetrachlorobis(1H-indazole)ruthenate (III)], mannitol, citric acid, and sodium citrate; wherein: sodium trans-[tetrachlorobis(1H-indazole)ruthenate (III)] is about 49.86 weight percent of the composition, mannitol is about 49.86 weight percent of the composition, citric acid is about 0.187 weight percent of the composition, and sodium citrate is about 0.093 weight percentage of the composition. In some such embodiments, the composition is a lyophilized powder.

[0044] One embodiment of the present invention provides a composition comprising sodium trans-[tetrachlorobis(1H-indazole)ruthenate (III)], mannitol, citric acid, and sodium citrate; wherein: sodium trans-[tetrachlorobis(1H-indazole)ruthenate (III)] is about 40 to about 60 weight percent of the composition, mannitol is about 40 to about 60 weight percent of the composition, citric acid is about 0.01 to about 0.5 weight percent of the composition, and sodium citrate is about 0.001 to about 0.25 weight percentage of the composition. In some such embodiments, the composition is a lyophilized powder.

[0045] One embodiment of the present invention provides a composition comprising sodium trans-[tetrachlorobis(1H-indazole)ruthenate (III)], mannitol, citric acid, and sodium citrate; wherein: sodium trans-[tetrachlorobis(1H-indazole)ruthenate (III)] is about 30 to about 70 weight percent of the composition, mannitol is about 30 to about 70 weight percent of the composition, citric acid is about 0.001 to about 1 weight percent of the composition, and sodium citrate is about 0.0001 to about 1 weight percentage of the composition. In some such embodiments, the composition is a lyophilized powder.

[0046] One embodiment of the present invention provides a composition comprising sodium trans-[tetrachlorobis(1H-indazole)ruthenate (III)], mannitol, citric acid, sodium citrate, and RuIIICl3(Hind)2(H2O); wherein: sodium trans-[tetrachlorobis(1H-indazole)ruthenate (III)] is about 49.86 weight percent of the composition, mannitol is about 49.86 weight percent of the composition, citric acid is about 0.187 weight percent of the composition, sodium citrate is about 0.093 weight percentage of the composition, and RuIIICl3(Hind)2(H2O) is not more than 0.5 weight percentage of the composition. In some such embodiments, the composition is a lyophilized powder.

[0047] One embodiment of the present invention provides a composition comprising sodium trans-[tetrachlorobis(1H-indazole)ruthenate (III)], mannitol, citric acid, sodium citrate, and RuIIICl3(Hind)2(H2O); wherein: sodium trans-[tetrachlorobis(1H-indazole)ruthenate (III)] is about 40 to about 60 weight percent of the composition, mannitol is about 40 to about 60 weight percent of the composition, citric acid is about 0.01 to about 0.5 weight percent of the composition, sodium citrate is about 0.001 to about 0.25 weight percentage of the composition, and RuIIICl3(Hind)2(H2O) is about 0 to about 0.5 weight percentage of the composition. In some such embodiments, the composition is a lyophilized powder.

[0048] One embodiment of the present invention provides a composition comprising sodium trans-[tetrachlorobis(1H-indazole)ruthenate (III)], mannitol, citric acid, sodium citrate, RuIIICl3(Hind)2(H2O), and cesium; wherein: sodium trans-[tetrachlorobis(1H-indazole)ruthenate (III)] is about 30 to about 70 weight percent of the composition, mannitol is about 30 to about 70 weight percent of the composition, citric acid is about 0.001 to about 1 weight percent of the composition, sodium citrate is about 0.0001 to about 1 weight percentage of the composition, RuIIICl3(Hind)2(H2O) is not more than 0.5 weight percentage of the composition, and cesium is not more than 0.25 weight percentage of the composition. In some such embodiments, the composition is a lyophilized powder.

[0049] One embodiment of the present invention provides a composition comprising sodium trans-[tetrachlorobis(1H-indazole)ruthenate (III)], mannitol, citric acid, sodium citrate, and cesium; wherein: sodium trans-[tetrachlorobis(1H-indazole)ruthenate (III)] is about 49.61 weight percent of the composition, mannitol is about 49.86 weight percent of the composition, citric acid is about 0.187 weight percent of the composition, sodium citrate is about 0.093 weight percentage of the composition and cesium is about 0.25 weight percentage of the composition. In some such embodiments, the composition is a lyophilized powder.

[0050] One embodiment of the present invention provides a composition comprising sodium trans-[tetrachlorobis(1H-indazole)ruthenate (III)], mannitol, citric acid, sodium citrate, and cesium; wherein: sodium trans-[tetrachlorobis(1H-indazole)ruthenate (III)] is about 40 to about 60 weight percent of the composition, mannitol is about 40 to about 60 weight percent of the composition, citric acid is about 0.01 to about 0.5 weight percent of the composition, sodium citrate is about 0.001 to about 0.25 weight percentage of the composition,and cesium is about 0.1 to about 0.5 weight percentage of the composition. In some such embodiments, the composition is a lyophilized powder.

[0051] One embodiment of the present invention provides a composition comprising sodium trans-[tetrachlorobis(1H-indazole)ruthenate (III)], mannitol, citric acid, sodium citrate, and cesium; wherein: sodium trans-[tetrachlorobis(1H-indazole)ruthenate (III)] is about 30 to about 70 weight percent of the composition, mannitol is about 30 to about 70 weight percent of the composition, citric acid is about 0.001 to about 1 weight percent of the composition, sodium citrate is about 0.0001 to about 1 weight percentage of the composition, and cesium is about 0.01 to about 1 weight percentage of the composition. In some such embodiments, the composition is a lyophilized powder.

[0052] One embodiment of the present invention provides a composition comprising sodium trans-[tetrachlorobis(1H-indazole)ruthenate (III)], mannitol, citric acid, sodium citrate, RuIIICl3(Hind)2(H2O), RuIIICl3(Hind)2(CH3CN), RuIIICl3(Hind)(HN=C(Me)ind), and cesium; wherein: sodium trans-[tetrachlorobis(1H-indazole)ruthenate (III)] about 46.61 weight percent of the composition, mannitol is about 49.86 weight percent of the composition, citric acid is about 0.187 weight percent of the composition, sodium citrate is about 0.093 weight percentage of the composition, RuIIICl3(Hind)2(H2O) is not more than 0.5 weight percentage of the composition, RuIIICl3(Hind)2(CH3CN) is not more than 1.25 weight percentage of the composition, RuIIICl3(Hind)(HN=C(Me)ind) is not more than 1.0 weight percentage of the composition, and cesium is not more than 0.25 weight percentage of the composition. In some such embodiments, the composition is a lyophilized powder.

[0053] One embodiment of the present invention provides a composition comprising sodium trans-[tetrachlorobis(1H-indazole)ruthenate (III)], mannitol, citric acid, sodium citrate, RuIIICl3(Hind)2(H2O), RuIIICl3(Hind)2(CH3CN), RuIIICl3(Hind)(HN=C(Me)ind), and cesium; wherein:sodium trans-[tetrachlorobis(1H-indazole)ruthenate (III)] about between 46.61 weight percent of the composition, mannitol is about 49.86 weight percent of the composition, citric acid is about 0.187 weight percent of the composition, sodium citrate is about 0.093 weight percentage of the composition, RuIIICl3(Hind)2(H2O) is not more than 0.5 weight percentage of the composition, RuIIICl3(Hind)2(CH3CN) is not more than 1.25 weight percentage of the composition, RuIIICl3(Hind)(HN=C(Me)ind) is not more than 1.0 weight percentage of the composition, and cesium is not more than 0.25 weight percentage of the composition. In some such embodiments, the composition is a lyophilized powder.

[0054] One embodiment of the present invention provides a composition comprising sodium trans-[tetrachlorobis(1H-indazole)ruthenate (III)], mannitol, citric acid, sodium citrate, RuIIICl3(Hind)2(H2O), RuIIICl3(Hind)2(CH3CN), RuIIICl3(Hind)(HN=C(Me)ind), and cesium; wherein: sodium trans-[tetrachlorobis(1H-indazole)ruthenate (III)] is about 40 to about 60 weight percent of the composition, mannitol is about 40 to about 60 weight percent of the composition, citric acid is about 0.01 to about 0.5 weight percent of the composition, sodium citrate is about 0.001 to about 0.25 weight percentage of the composition, RuIIICl3(Hind)2(H2O) is not more than about 0.5 weight percentage of the composition, RuIIICl3(Hind)2(CH3CN) is not more than about 1.25 weight percentage of the composition, RuIIICl3(Hind)(HN=C(Me)ind) is not more than about 1.0 weight percentage of the composition, and cesium is not more than 0.25 percentage of the composition. In some such embodiments, the composition is a lyophilized powder.

[0055] One embodiment of the present invention provides a composition comprising sodium trans-[tetrachlorobis(1H-indazole)ruthenate (III)], mannitol, citric acid, sodium citrate, RuIIICl3(Hind)2(H2O), RuIIICl3(Hind)2(CH3CN), RuIIICl3(Hind)(HN=C(Me)ind), and cesium; wherein: sodium trans-[tetrachlorobis(1H-indazole)ruthenate (III)] is about 30 to about 70 weight percent of the composition,mannitol is about 30 to about 70 weight percent of the composition, citric acid is about 0.001 to about 1 weight percent of the composition, sodium citrate is about 0.0001 to about 1 weight percentage of the composition, RuIIICl3(Hind)2(H2O) is not more than about 0.5 weight percentage of the composition, RuIIICl3(Hind)2(CH3CN) is not more than about 1.25 weight percentage of the composition, RuIIICl3(Hind)(HN=C(Me)ind) is not more than about 1.0 weight percentage of the composition, and cesium is not more than 0.25 percentage of the composition. In some such embodiments, the composition is a lyophilized powder.

[0056] One embodiment of the present invention provides a composition comprising sodium trans-[tetrachlorobis(1H-indazole)ruthenate (III)], mannitol, citric acid, sodium citrate, RuIIICl3(Hind)2(H2O), RuIIICl3(Hind)2(CH3CN), RuIIICl3(Hind)(HN=C(Me)ind), and cesium; wherein: sodium trans-[tetrachlorobis(1H-indazole)ruthenate (III)] is about 20 to about 80 weight percent of the composition, mannitol is about 20 to about 80 weight percent of the composition, citric acid is about 0.0001 to about 5 weight percent of the composition, sodium citrate is about 0.00001 to about 5 weight percentage of the composition, RuIIICl3(Hind)2(H2O) is not more than about 0.5 weight percentage of the composition, RuIIICl3(Hind)2(CH3CN) is not more than about 1.25 weight percentage of the composition, RuIIICl3(Hind)(HN=C(Me)ind) is not more than about 1.0 weight percentage of the composition, and cesium is not more than 0.25 percentage of the composition. In some such embodiments, the composition is a lyophilized powder.

[0057] In some embodiments, the present invention provides a unit dosage form comprising a formulation or composition described herein. The expression "unit dosage form" as used herein refers to a physically discrete unit of a provided formulation appropriate for the subject to be treated. It will be understood, however, that the total daily usage of provided formulation will be decided by the attending physician within the scope of sound medical judgment. The specific effective dose level for any particular subject or organism will dependupon a variety of factors including the disorder being treated and the severity of the disorder; activity of specific active agent employed; specific formulation employed; age, body weight, general health, sex and diet of the subject; time of administration, and rate of excretion of the specific active agent employed; duration of the treatment; drugs and / or additional therapies used in combination or coincidental with specific compound(s) employed, and like factors well known in the medical arts.

[0058] Compositions of the present invention can be provided as a unit dosage form. In some embodiments, a vial comprising sodium trans-[tetrachlorobis(1H-indazole)ruthenate (III)], mannitol, citric acid, sodium citrate is a unit dosage form.

[0059] In some embodiments, the present invention a vial comprising sodium trans- [tetrachlorobis(1H-indazole)ruthenate (III)], mannitol, citric acid, sodium citrate, and cesium is a unit dosage form.

[0060] In some embodiments, the present invention a vial comprising sodium trans- [tetrachlorobis(1H-indazole)ruthenate (III)], mannitol, citric acid, sodium citrate, RuIIICl3(Hind)2(H2O), RuIIICl3(Hind)2(CH3CN), RuIIICl3(Hind)(HN=C(Me)ind),and cesium is a unit dosage form.

[0061] Still further encompassed by the invention are pharmaceutical packs and / or kits comprising compositions described herein, or a unit dosage form comprising a provided composition, and a container (e.g., a foil or plastic package, or other suitable container). Optionally instructions for use are additionally provided in such kits.

[0062] In some embodiments, the present invention can be provided as a unit dosage form. Indeed, a vial comprising sodium trans-[tetrachlorobis(1H-indazole)ruthenate (III)], mannitol, citric acid, sodium citrate is a unit dosage form depicted in Table 3 Table 3: Pharmaceutical Components

[0063] In some embodiments, the pharmaceutical components described in Table 3 further comprise cesium; wherein: cesium is not more than 0.25 weight percentage of the composition.

[0064] In some embodiments, the pharmaceutical components described in Table 3 further comprise cesium, RuIIICl3(Hind)2(H2O), RuIIICl3(Hind)2(CH3CN), and RuIIICl3(Hind)(HN=C(Me)ind); wherein: cesium is not more than about 0.25 weight percentage of the composition, RuIIICl3(Hind)2(H2O) is not more than about 0.5 weight percentage of the composition, RuIIICl3(Hind)2(CH3CN) is not more than about 1.25 weight percentage of the composition, and RuIIICl3(Hind)(HN=C(Me)ind) is not more than about 1.0 weight percentage of the composition.

[0065] In some embodiments, the pharmaceutical composition is selected from those in Table 4: Table 4: Pharmaceutical Component Ranges

[0066] In some embodiments, the pharmaceutical components described in Table 4 further comprise cesium; wherein: cesium is not more than 0.25 weight percentage of the composition.

[0067] In some embodiments, the pharmaceutical components described in Table 4 further comprise cesium, RuIIICl3(Hind)2(H2O), RuIIICl3(Hind)2(CH3CN), and RuIIICl3(Hind)(HN=C(Me)ind);wherein: cesium is not more than about 0.25 weight percentage of the composition, RuIIICl3(Hind)2(H2O) is not more than about 0.5 weight percentage of the composition, RuIIICl3(Hind)2(CH3CN) is not more than about 1.25 weight percentage of the composition, and RuIIICl3(Hind)(HN=C(Me)ind) is not more than about 1.0 weight percentage of the composition.

[0068] In some embodiments, the present invention can be provided as a unit dosage form. Indeed, a vial comprising sodium trans-[tetrachlorobis(1H-indazole)ruthenate (III)], mannitol, citric acid, sodium citrate is a unit dosage form depicted in Table 5: Table 5: Pharmaceutical Components

[0069] In some embodiments, the pharmaceutical components described in Table 5 further comprise cesium; wherein: cesium is not more than 0.25 weight percentage of the composition.

[0070] In some embodiments, the pharmaceutical components described in Table 5 further comprise cesium, RuIIICl3(Hind)2(H2O), RuIIICl3(Hind)2(CH3CN), and RuIIICl3(Hind)(HN=C(Me)ind); wherein: cesium is not more than about 0.25 weight percentage of the composition, RuIIICl3(Hind)2(H2O) is not more than about 0.5 weight percentage of the composition, RuIIICl3(Hind)2(CH3CN) is not more than about 1.25 weight percentage of the composition, and RuIIICl3(Hind)(HN=C(Me)ind) is not more than about 1.0 weight percentage of the composition.

[0071] In some embodiments, the pharmaceutical composition is selected from those in Table 6: Co sod inda Ma Citr Sod0.1012

[0072] In some embodiments, the pharmaceutical components described in Table 6 further comprise cesium; wherein: cesium is not more than 0.25 weight percentage of the composition.

[0073] In some embodiments, the pharmaceutical components described in Table 6 further comprise cesium, RuIIICl3(Hind)2(H2O), RuIIICl3(Hind)2(CH3CN), and RuIIICl3(Hind)(HN=C(Me)ind); wherein: cesium is not more than about 0.25 weight percentage of the composition, RuIIICl3(Hind)2(H2O) is not more than about 0.5 weight percentage of the composition, RuIIICl3(Hind)2(CH3CN) is not more than about 1.25 weight percentage of the composition, and RuIIICl3(Hind)(HN=C(Me)ind) is not more than about 1.0 weight percentage of the composition.

[0074] In some embodiments, the pharmaceutical components are as described in any of Tables 3-6, and further comprise cesium. In some embodiments, cesium is present in an amount of about 0.001, 0.002, 0.003, 0.004, 0.005, 0.006, 0.007, 0.008, 0.009, 0.010, 0.015, 0.020, 0.025, 0.030, 0.035, 0.040, 0.045, 0.050, 0.055, 0.060, 0.065, 0.070, 0.075, 0.080, 0.085, 0.090, 0.095, 0.10, 0.15, 0.20, 0.25, 0.30, 0.35, 0.40, 0.45, 0.50, 0.55, 0.60, 0.65, 0.70, 0.75, 0.80, 0.85, 0.90, 0.95, or 1.0 weight percentage of the composition.

[0075] In some embodiments, the effective amount of BOLD-100 may be effective to reduce the amount of GRP78 in cancer cells following administration of BOLD-100, either alone or in combination.

[0076] According to one embodiment of the present invention provides a method for treating cancer in a patient in need thereof, comprising the steps of: 1) identifying the patient as having a mutation in a TP53 gene; 2) administering to the patient a chemotherapy agent; 2) subsequently administering BOLD-100, or a pharmaceutically acceptable composition thereof; to the patient; and 3) optionally repeating steps 1 and 2.

[0077] In certain embodiments, the BOLD-100, or a pharmaceutically acceptable composition thereof, is administered before, after or simultaneously with a chemotherapeutic agent, for example 1 day before or after the chemotherapy agent, 1 week before or after the chemotherapy agent, between 1 and seven days before or after the chemotherapy agent. In certain embodiments, the BOLD-100, or a pharmaceutically acceptable composition thereof, and the chemotherapy agent are administered within about 20-28 hours of each other, or within about 22-26 hours of each other, or within about 24 hours of each other.

[0078] A titratable dosage may for example be adapted to allow a patient to take the medication in doses smaller than the unit dose, wherein a "unit dose" is defined as the maximum dose of medication that can be taken at any one time or within a specific dosage period. Titration of doses will allow different patients to incrementally increase the dose until they feel that the medication is efficacious, as not all patients will require the same dose to achieve the same benefits. A person with a larger build or faster metabolism may require larger doses to achieve the same effect as another with a smaller build or slower metabolism. Therefore, a titratable dosage has advantages over a standard dosage form.

[0079] In select embodiments, formulations may be adapted to be delivered in such a way as to target one or more of the following: sublingual, buccal, oral, rectal, nasal, parenteral and via the pulmonary system. Formulations may for example be in one or more of the following forms: gel, gel spray, tablet, liquid, capsule, by injection, or for vaporization.

[0080] Conventional pharmaceutical practice may be employed to provide suitable formulations or compositions to administer the formulations to subjects. Routes of administration may for example include parenteral, intravenous, intradermal, subcutaneous, intramuscular,intracranial, intraorbital, ophthalmic, intraventricular, intracapsular, intraspinal, intrathecal, intracisternal, intraperitoneal, intranasal, inhalational, aerosol, topical, sublingual, or oral administration. Therapeutic formulations may be in the form of liquid solutions or suspensions; for oral administration, formulations may be in the form of tablets or capsules; for intranasal formulations, in the form of powders, nasal drops, or aerosols; and for sublingual formulations, in the form of drops, aerosols or tablets.

[0081] Methods well known in the art for making formulations are found in, for example, “Remington: The Science and Practice of Pharmacy” (21st edition), ed. David Troy, 2006, Lippincott Williams & Wilkins. Formulations for parenteral administration may, for example, contain excipients, sterile water, or saline, polyalkylene glycols such as polyethylene glycol, oils of vegetable origin, or hydrogenated napthalenes. Biocompatible, biodegradable lactide polymer, lactide / glycolide copolymer, or polyoxyethylene-polyoxypropylene copolymers may be used to control the release of the compounds. Other potentially useful parenteral delivery systems include ethylene-vinyl acetate copolymer particles, osmotic pumps, implantable infusion systems, and liposomes. Formulations for inhalation may contain excipients, for example, lactose, or may be aqueous solutions containing, for example, polyoxyethylene-9-lauryl ether, glycocholate and deoxycholate, or may be oily solutions for administration in the form of nasal drops, or as a gel.

[0082] Pharmaceutical compositions of the present invention may be in any form which allows for the composition to be administered to a patient. For example, the composition may be in the form of a solid, liquid or gas (aerosol). Pharmaceutical composition of the invention are formulated so as to allow the active ingredients contained therein to be bioavailable upon administration of the composition to a patient. Compositions that will be administered to a patient may take the form of one or more dosage units, where for example, a tablet, capsule or cachet may be a single dosage unit, and a container of the compound in aerosol form may hold a plurality of dosage units.

[0083] Materials used in preparing the pharmaceutical compositions should be pharmaceutically pure and non-toxic in the amounts used. The inventive compositions may include one or more compounds (active ingredients) known for a particularly desirable effect. It will be evident to those of ordinary skill in the art that the optimal dosage of the active ingredient(s) in the pharmaceutical composition will depend on a variety of factors. Relevant factors include, without limitation, the type of subject (e.g., human), the particular form of the active ingredient, the manner of administration and the composition employed.

[0084] In general, the pharmaceutical composition includes a formulation of the present invention as described herein, in admixture with one or more carriers. The carrier(s) may be particulate, so that the compositions are, for example, in tablet or powder form. The carrier(s) may be liquid, with the compositions being, for example, an oral syrup or injectable liquid. In addition, the carrier(s) may be gaseous, so as to provide an aerosol composition useful in, e.g., inhalatory administration.

[0085] When intended for oral administration, the composition is preferably in either solid or liquid form, where semi-solid, semi-liquid, suspension and gel forms are included within the forms considered herein as either solid or liquid.

[0086] As a solid formulation for oral administration, the composition may be formulated into a powder, granule, compressed tablet, pill, capsule, cachet, chewing gum, wafer, lozenges, or the like form. Such a solid composition will typically contain one or more inert diluents or edible carriers. In addition, one or more of the following adjuvants may be present: binders such as syrups, acacia, sorbitol, polyvinylpyrrolidone, carboxymethylcellulose, ethyl cellulose, microcrystalline cellulose, gum tragacanth or gelatin, and mixtures thereof; excipients such as starch, lactose or dextrins, disintegrating agents such as alginic acid, sodium alginate, Primogel, corn starch and the like; lubricants such as magnesium stearate or Sterotex; fillers such as lactose, mannitols, starch, calcium phosphate, sorbitol, methylcellulose, and mixtures thereof; lubricants such as magnesium stearate, high molecular weight polymers such as polyethylene glycol, high molecular weight fatty acids such as stearic acid, silica, wetting agents such as sodium lauryl sulfate, glidants such as colloidal silicon dioxide; sweetening agents such as sucrose or saccharin, a flavoring agent such as peppermint, methyl salicylate or orange flavoring, and a coloring agent. When the composition is in the form of a capsule, e.g., a gelatin capsule, it may contain, in addition to materials of the above type, a liquid carrier such as polyethylene glycol or a fatty oil.

[0087] The formulation may be in the form of a liquid, e.g., an elixir, syrup, solution, aqueous or oily emulsion or suspension, or even dry powders which may be reconstituted with water and / or other liquid media prior to use. The liquid may be for oral administration or for delivery by injection, as two examples. When intended for oral administration, preferred compositions contain, in addition to the present compounds, one or more of a sweetening agent, thickening agent, preservative (e.g., alkyl p-hydoxybenzoate), dye / colorant and flavor enhancer (flavorant). In a composition intended to be administered by injection, one or more of a surfactant, preservative(e.g., alkyl p-hydroxybenzoate), wetting agent, dispersing agent, suspending agent (e.g., sorbitol, glucose, or other sugar syrups), buffer, stabilizer and isotonic agent may be included. The emulsifying agent may be selected from lecithin or sorbitol monooleate.

[0088] The liquid pharmaceutical formulations of the invention, whether they be solutions, suspensions or other like form, may include one or more of the following adjuvants: sterile diluents such as water for injection, saline solution, preferably physiological saline, Ringer’s solution, isotonic sodium chloride, fixed oils such as synthetic mono or digylcerides which may serve as the solvent or suspending medium, polyethylene glycols, glycerin, propylene glycol or other solvents; antibacterial agents such as benzyl alcohol or methyl paraben; antioxidants such as ascorbic acid or sodium bisulfite; chelating agents such as ethylenediaminetetraacetic acid; buffers such as acetates, citrates or phosphates and agents for the adjustment of tonicity such as sodium chloride or dextrose. The parenteral preparation can be enclosed in ampoules, disposable syringes or multiple dose vials made of glass or plastic. Physiological saline is a preferred adjuvant. An injectable pharmaceutical composition is preferably sterile.

[0089] The pharmaceutical formulation may be intended for topical administration, in which case the carrier may suitably comprise a solution, emulsion, ointment, cream or gel base. The base, for example, may comprise one or more of the following: petrolatum, lanolin, polyethylene glycols, bee wax, mineral oil, diluents such as water and alcohol, and emulsifiers and stabilizers. Thickening agents may be present in a pharmaceutical composition for topical administration. If intended for transdermal administration, the composition may include a transdermal patch or iontophoresis device.

[0090] The formulation may be intended for rectal administration, in the form, e.g., of a suppository which will melt in the rectum and release the drug. The composition for rectal administration may contain an oleaginous base as a suitable nonirritating excipient. Such bases include, without limitation, lanolin, cocoa butter and polyethylene glycol. Low-melting waxes are preferred for the preparation of a suppository, where mixtures of fatty acid glycerides and / or cocoa butter are suitable waxes. The waxes may be melted, and the aminocyclohexyl ether compound is dispersed homogeneously therein by stirring. The molten homogeneous mixture is then poured into convenient sized molds, allowed to cool and thereby solidify.

[0091] The formulation may include various materials which modify the physical form of a solid or liquid dosage unit. For example, the composition may include materials that form a coating shell around the active ingredients. The materials which form the coating shell aretypically inert, and may be selected from, for example, sugar, shellac, and other enteric coating agents. Alternatively, the active ingredients may be encased in a gelatin capsule or cachet.

[0092] The pharmaceutical formulation may consist of gaseous dosage units, e.g., it may be in the form of an aerosol. The term aerosol is used to denote a variety of systems ranging from those of colloidal nature to systems consisting of pressurized packages. Delivery may be by a liquefied or compressed gas or by a suitable pump system which dispenses the active ingredients. Aerosols of compounds of the invention may be delivered in single phase, bi-phasic, or tri-phasic systems in order to deliver the active ingredient(s). Delivery of the aerosol includes the necessary container, activators, valves, subcontainers, and the like, which together may form a kit.

[0093] Some biologically active compounds may be in the form of the free base or in the form of a pharmaceutically acceptable salt such as the hydrochloride, sulfate, phosphate, citrate, fumarate, methanesulfonate, acetate, tartrate, maleate, lactate, mandelate, salicylate, succinate and other salts known in the art. The appropriate salt would be chosen to enhance bioavailability or stability of the compound for the appropriate mode of employment (e.g., oral or parenteral routes of administration).

[0094] The present invention also provides kits that contain a pharmaceutical formulation, together with instructions for the use of the formulation. Preferably, a commercial package will contain one or more unit doses of the formulation. Formulations which are light and / or air sensitive may require special packaging and / or formulation. For example, packaging may be used which is opaque to light, and / or sealed from contact with ambient air, and / or formulated with suitable coatings or excipients.

[0095] The formulations of the invention can be provided alone or in combination with other compounds (for example, small molecules, nucleic acid molecules, peptides, or peptide analogues), in the presence of a carrier or any pharmaceutically or biologically acceptable carrier. As used herein “pharmaceutically acceptable carrier” or “excipient” includes any and all solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonic and absorption delaying agents, and the like that are physiologically compatible. The carrier can be suitable for any appropriate form of administration. Pharmaceutically acceptable carriers generally include sterile aqueous solutions or dispersions and sterile powders. Supplementary active compounds can also be incorporated into the formulations.

[0096] An “effective amount” of a formulation according to the invention includes a therapeutically effective amount or a prophylactically effective amount. A “therapeuticallyeffective amount” refers to an amount effective, at dosages and for periods of time necessary, to achieve the desired therapeutic result. A therapeutically effective amount of a formulation may vary according to factors such as the disease state, age, sex, and weight of the individual, and the ability of the compound to elicit a desired response in the individual. Dosage regimens may be adjusted to provide the optimum therapeutic response. A therapeutically effective amount may also be one in which any toxic or detrimental effects of the formulation or active compound are outweighed by the therapeutically beneficial effects. A “prophylactically effective amount” refers to an amount effective, at dosages and for periods of time necessary, to achieve the desired prophylactic result. Typically, a prophylactic dose is used in subjects prior to or at an earlier stage of disease, so that a prophylactically effective amount may be less than a therapeutically effective amount. For any particular subject, the timing and dose of treatments may be adjusted over time (e.g., timing may be daily, every other day, weekly, monthly) according to the individual need and the professional judgment of the person administering or supervising the administration of the compositions. EXAMPLES

[0097] As illustrated in the following Examples, BOLD-100 in combination with a FOLFOX chemotherapy regimen provides improved overall survival (OS) and progression free survival (PFS) in metastatic colorectal cancer (mCRC) patients with cancers having TP53 mutations (TP53MT). TP53 gain-of-function (GOF) mutations were associated with the greatest improved OS and PFS compared to the TP53 non-GOF and TP53 wildtype patients. These Examples according illustrate that TP53 mutations, and especially TP53 GOF mutations, are predictive for improved OS and PFS when treated with BOLD-100.

[0098] As illustrated herein, in mCRC patients (n=42) TP53 mutation was associated with improved OS and PFS. Gain-of-function mutations were associated with a particularly improved OS and PFS. Results were similar when only looking in the 3rdline and beyond subset (n=38).

[0099] Overall, advanced heavily pre-treated metastatic colorectal cancer patients treated with BOLD-100 and FOLFOX showed a median PFS of 4.2 months, OS of 8.6 months, ORR of 11%, and DCR of 77%, substantially higher than standard-of-care data (Bayer’s Stivarga® and Taiho’s Lonsurf®) for a similar patient population which showed a median PFS of 2.0 months, OS of 7.1 months, ORR of only 1.5%, and DCR of 44%, with some of the most remarkable outcomes for BOLD-100 in late-line patients that had previously failed on FOLFOX alone.BOLD-100 in combination with FOLFOX was well-tolerated, with patients remaining on therapy for up to 18 treatment cycles. Against this backdrop of an overall very effective therapy, the discovery that positive TP53 mutational status is a strong predictive indicator of clinical efficacy is especially meaningful. Example 1: Patients

[0100] Results herein are obtained from a cohort of 42 patients treated with BOLD-100 in combination with FOLFOX (folinic acid / leucovorin, fluorouracil, and oxaliplatin) chemotherapy for metastatic colorectal cancer (mCRC).42 patients with advanced metastatic colorectal cancer median age 62 years were treated.35 were evaluable for efficacy endpoints. Patients received a median of 4 prior systemic therapies (range 1-8), 38 had received prior FOLFOX and 41 (98%) were enrolled with stage IV disease. Median number of cycles completed was 6 (range 1-18). Median PFS was 4.2 [3.0, 5.9] months, median OS 8.6 [6.0, 13] months, and ORR 11% [4, 25] compared to the historical benchmark of 2.0 months, 7.1 months, and 1.6% respectively for similar patients treated with approved standard of care. One patient (2.8%) achieved a complete response, three patients (8.6%) achieved a partial response, and 23 patients (65.7%) had stable disease for an overall disease control rate of 77% (27 / 35 [61%, 89%]). This compares favorably to the historical control 44%.

[0101] Patients were administered BOLD-100+mFOLFOX6 on day 1 of each 14-day cycle. Four cohorts are treated at the BOLD-100 dose of 625 mg / m2with mFOLFOX6 until progressive disease or unacceptable toxicity. The mFOLFOX6 drug regimen is as follows: • oxaliplatin 85 mg / m² IV in 500mL D5W over 120 minutes, Day 1; • leucovorin 400 mg / m² IV diluted in D5W, over 120 minutes (concurrently with oxaliplatin), Day 1; • fluorouracil 400 mg / m² IV bolus, after leucovorin, Day 1; • THEN; • fluorouracil 2400 mg / m² IV continuous infusion over 46 hours (single dose).

[0102] Plasma was collected from each patient during the first 2 cycles of treatment. Circulating tumour DNA (ctDNA) analysis was completed on the pooled samples using IMBDx’s Alpha-100 profiling. Kaplan-Meier survival and Cox proportional hazards models were used to assess patient outcome with respect to gene mutation status.Example 2: Univariate Hazard Ratios

[0103] In all mCRC patients, presence of a TP53 mutation was associated with a hazard ratio below 1 for both overall survival (OS; p = 0.089) and progression free survival (PFS; p = 0.0077). Example 3: Multivariate Hazard Ratio for TP53 Mutational Status

[0104] A multivariate hazard ratio model with all mCRC patients was completed with TP53 and standard clinical characteristics (age, sex, race, etc.). TP53 mutation had a significant hazard ratio <1 for both overall survival and progression-free survival. Example 4: Kaplan-Meier Survival Curves in TP53 mCRC Patients

[0105] As illustrated in Figure 1, Kaplan-Meier Survival Curves show patients with TP53 mutations had longer overall survival (Figure 1A) and progression-free survival (Figure 1B). Example 5: Kaplan-Meier Curves in mCRC Patients with GOF Mutations

[0106] As illustrated in Figure 2, TP53 “Gain-of-Function” (GOF) mutations provide an additional degree of predictive value for response to BOLD-100 treatment compared to non- GOF mutations. In this context, GOF mutations are generally defined as mutations that not only inhibit TP53 cancer suppression function, but also confer additional pro-cancerous phenotypes. The following mutations were defined as GOF mutations for the TP53 gene: R175H, G245S, R248Q, R248W, R249S, R273H, R273L, and R282W. In total, 11 out of the 32 TP53MT patients had at least one GOF mutation.

[0107] Kaplan-Meier Survival Curves show patients with TP53 GOF mutations had longer OS (Figure 2A) and PFS (Figure 2B) than patients with TP53 non-GOF mutations. Both the TP53 GOF and TP53 non-GOF mutation cohorts had improved OS (Figure 2A) and PFS (Figure 2B) compared to WT patients. Example 6: Kaplan-Meier Curves with GOF Mutations by Sidedness

[0108] TP53 mutations are often more present in left sided mCRC tumours, which are understood to have a better prognosis. When looking at a subset of patients only with left-sided tumours, patients with TP53 mutations had improved OS (Figure 3A) and PFS (Figure 3B).These results illustrate that the improved response of patients with TP53 mutations to BOLD- 100 treatment is independent of tumour sidedness. REFERENCES

[0109] P. Heffeter, M. Pongratz, E. Steiner, P. Chiba, M. A. Jakupec, L. Elbling, B. Marian, W. Körner, F. Sevelda, M. Micksche, B. K. Keppler and W. Berger, Intrinsic and Acquired Forms of Resistance against the Anticancer Ruthenium Compound KP1019 [Indazolium trans-[tetrachlorobis(1H-indazole)ruthenate (III)] (FFC14A), Journal of Pharmacology and Experimental Therapeutics January 1, 2005, 312 (1) 281-289.

[0110] Venderbosch S, Nagtegaal ID, Maughan TS, et al. Clin Cancer Res. 2014;20(20):5322-30.

[0111] Lochhead P, Kuchiba A, Imamura Y, et al. J Natl Cancer Inst. 2013;105(15):1151-6.

[0112] Kopetz, S. et al. Encorafenib, Binimetinib, and Cetuximab in BRAF V600E- Mutated Colorectal Cancer. N Engl J Med.381, 1632-1643 (2019).

[0113] Guinney, J. et al. The consensus molecular subtypes of colorectal cancer. Nat Med.21, 1350-1356 (2015).

[0114] Medico, E. et al. The molecular landscape of colorectal cancer cell lines unveils clinically actionable kinase targets. Nature communications.6, 7002 (2015).

[0115] Burris, H.A. et al. Safety and activity of IT-139, a ruthenium-based compound, in patients with advanced solid tumours: a first-in-human, open-label, dose-escalation phase I study with expansion cohort. ESMO open.1, e000154 (2016).

[0116] Although various embodiments of the invention are disclosed herein, many adaptations and modifications may be made within the scope of the invention in accordance with the common general knowledge of those skilled in this art. Such modifications include the substitution of known equivalents for any aspect of the invention in order to achieve the same result in substantially the same way. Terms such as “exemplary” or “exemplified” are used herein to mean “serving as an example, instance, or illustration.” Any implementation described herein as “exemplary” or “exemplified” is accordingly not to be construed as necessarily preferred or advantageous over other implementations, all such implementations being independent embodiments. Unless otherwise stated, numeric ranges are inclusive of the numbers defining the range, and numbers are necessarily approximations to the given decimal.The word "comprising" is used herein as an open-ended term, substantially equivalent to the phrase "including, but not limited to", and the word "comprises" has a corresponding meaning. As used herein, the singular forms "a", "an" and "the" include plural referents unless the context clearly dictates otherwise. Thus, for example, reference to "a thing" includes more than one such thing. Citation of references herein is not an admission that such references are prior art to the present invention. Any priority document(s) and all publications, including but not limited to patents and patent applications, cited in this specification, and all documents cited in such documents and publications, are hereby incorporated herein by reference as if each individual publication were specifically and individually indicated to be incorporated by reference herein and as though fully set forth herein. The invention includes all embodiments and variations substantially as hereinbefore described and with reference to the examples and drawings.

[0117] In some embodiments, the invention excludes steps that involve medical or surgical treatment.

Claims

CLAIMS 1. A method of treating a cancer with a mutation in a TP53 gene in a human patient in need thereof, comprising: identifying the cancer in the patient as having the TP53 mutation; and, administering an effective amount of sodium trans-[tetrachloridobis(1H- indazole)ruthenate(III)] to treat the cancer.

2. The method of claim 1, wherein the TP53 mutation mediates a substitution, deletion or insertion at an altered amino acid position in a p53 protein encoded by the TP53 gene.

3. The method of claim 2, wherein the altered amino acid position is at one or more of positions 175, 245, 248, 249, 273, or 282 in the sequence set out in SEQ ID NO:

2.

4. The method of claim 3, wherein the TP53 mutation mediates a substitution in the p53 protein that is one or more of R175A, R175C, R175G, R175H, R175L R175P, G245A, G245C, G245D, G245F, G245R, G245S, G245V, R248E, R248G, R248L, R248P, R248Q, R248W, R249G, R249K, R249M, R249S, R249T, R249W, R273C, R273H, R273L, R273P, R273S, R273W, R273Y, R282D, R282G, R282H, R282Q or R282W.

5. The method of claim 1, wherein the mutation is a gain-of-function (GOF) mutation in the TP53 gene.

6. The method of claim 5, wherein the GOF mutation mediates a substitution in the p53 protein that is one or more of S121X, C124X, V157X, A161X, R175X, L194X, T211X, F212X, H214X, G245X, R248X, R249X, R273X, C277X, R280X, D281X, R282X, R290X, K291X, K292X, or G360X, where X is any substitute amino acid.

7. The method of claim 6, wherein the substitution in the p53 protein is one or more of S121A, S121C, S121F, C124S, V157S, A161T, R175H, L194R, T211A, F212Y, H214R, G245A, G245C, G245D, G245S, R248G, R249W, R273H, C277S, C277Y, R280K, D281G, R282Q, R290G, K291E, K291Q, K291T, K291I, K292T, or G360V.

8. The method of claim 7, wherein the substitution in the p53 protein is one or more of R175H, G245S, R248Q, R248W, R249S, R273H, R273L, or R282W.

9. The method of any one of claims 1-8, wherein the cancer is a colorectal, gastric, pancreatic, esophageal, breast or biliary tract cancer.

10. The method of any one of claims 1-8, wherein the cancer is a metastatic colorectal cancer (mCRC).

11. The method of any one of claims 1-10, wherein the cancer is a stage II, III or IV cancer.

12. The method of any one of claims 1-11, wherein sodium trans- [tetrachloridobis(1H-indazole)ruthenate(III)] is administered in combination with one or more additional therapeutic agent to treat the cancer.

13. The method of claim 12, wherein the additional therapeutic agent comprises a platinum-based chemotherapeutic; an antimetabolite; an anthracycline; an EGFR targeted therapeutic; a poly (ADP-ribose) polymerase PARP targeted therapeutic; a vascular endothelial growth factor (VEGF) or vascular endothelial growth factor receptor (VEGFR) targeted therapeutic, or an immunotherapeutic.

14. The method of claim 13, wherein the platinum-based chemotherapeutic comprises cisplatin, carboplatin or oxaliplatin.

15. The method of claim 13, wherein the antimetabolite comprises methotrexate, 5- FU, aminopterin, cytarabine, thioguanine, or mercaptopurine.

16. The method of claim 13, wherein the anthracycline comprises daunorubicin, doxorubicin, epirubicin, idarubicin, mitoxantrone or valrubicin.

17. The method of claim 13, wherein the EGFR targeted therapeutic comprises afatinib, gefitnib, amivantamab-vmjw, osimertinib, erlotinib, or dacomitinib.

18. The method of claim 13, wherein the PARP targeted therapeutic olaparib, rucaparib, or niraparib.

19. The method of claim 13, wherein VEGF / VEGFR targeted therapeutic comprises sunitinib, pazopanib, foretinib, sorafenib, tivozanib, anlotinib, axitinib, golvatinib, fresolimumab, trastuzumab, ramucirumab, tanibirumab, sym004, bevacizumab, aflibercept, or sevacizumab.

20. The method of any one of claims 1-8, wherein the cancer is a colorectal cancer (CRC) or metastatic colorectal cancer (mCRC) and sodium trans-[tetrachlorobis(1H- indazole)ruthenate (III)] is administered in combination with a FOLFOX treatment.

21. The method of any one of claims 1-8, wherein the cancer is a colorectal cancer (CRC) or metastatic colorectal cancer (mCRC) and sodium trans-[tetrachlorobis(1H- indazole)ruthenate (III)] is administered in combination with irinotecan.

22. The method of any one of claims 1-8, wherein the cancer is a colorectal cancer (CRC) or metastatic colorectal cancer (mCRC) and sodium trans-[tetrachlorobis(1H- indazole)ruthenate (III)] is administered in combination with a VEGF inhibitor, an EGFR inhibitor, a PARP inhibitor.

23. The method of any one of claims 1-22, wherein the treatment is a first line treatment.

24. The method of any one of claims 1-22, wherein the treatment is second, third, fourth, or subsequent line treatment.

25. The method of any one of claims 1-22, wherein the treatment is an adjuvant treatment.

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