Thiostrepton dosing regimen

Abstract

Disclosed herein are methods of administering thiostrepton or pharmaceutical compositions thereof, and methods of treating cancer.

Description

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS

[0002] This application claims the benefit of priority to U.S. Provisional Patent Application No. 63 / 404,728, filed on September 8, 2022, and U.S. Provisional Patent Application No. 63 / 403,553, filed on September 2, 2022, each of which is hereby incorporated by reference in its entirety. Background Art

[0003] Thiostrepton is a cyclic oligopeptide antibiotic, which also has other names such as Bryamycin, Thiactin, Alaninamide, HR4S203Y18, etc. Thiostrepton has the following structure:

[0004]

[0005] or a pharmaceutically acceptable salt thereof. Recent studies have shown that thiostrepton, in addition to its antibiotic properties, also has promising anticancer activity. There remains a need for safe and effective methods of administering thiostrepton to treat cancer. Summary of the invention

[0006] In certain aspects, provided herein are methods of treating cancer comprising administering thiostrepton to a subject in need thereof once per week in a single dose of about 50 mg to about 500 mg, thereby treating the cancer.

[0007] In certain aspects, provided herein are pharmaceutical compositions comprising thiostrepton and at least one pharmaceutically acceptable carrier, wherein the pharmaceutical composition comprises about 50 mg to about 500 mg of thiostrepton.

[0008] In certain embodiments, the present disclosure provides for administration of thiostrepton or any pharmaceutical composition comprising thiostrepton disclosed herein.

[0009] Also provided herein is a method for treating cancer, the method comprising administering thiostrepton as described herein or a pharmaceutical composition comprising thiostrepton to a subject in need thereof. In certain preferred embodiments, cancer is malignant mesothelioma. In certain embodiments, thiostrepton is administered intraperitoneally, intrapleurally, subcutaneously or intratumorally. In certain embodiments, thiostrepton is administered intraperitoneally, for example, using an indwelling intraperitoneal catheter (IPC) or a drainage port catheter. In certain embodiments, the method further includes removing liquid (such as pleural effusion or ascites from the peritoneal cavity) from the body cavity before administering thiostrepton, for example to dryness. In some embodiments, thiostrepton is administered once a week for at least 3 weeks. In a preferred embodiment, a single dose of thiostrepton is administered once a week with about 90 mg to about 450 mg, such as about 90 mg, about 180 mg, about 270 mg, about 360 mg and about 450 mg. BRIEF DESCRIPTION OF THE DRAWINGS

[0010] Figure 1A The nucleus and mitochondria of a normal mesothelial cell are depicted.

[0011] Figure 1B The nucleus and mitochondria of a malignant mesothelial cell are depicted.

[0012] Figure 2A EC of thiostrepton in normal mesothelial cell lines and various mesothelioma cell lines (different BAP1 expression) are plotted. 50 .

[0013] Figure 2B It is shown that knockdown of PRX3 using siRNA significantly reduced MM (malignant melanoma, HM cell line-pleural biphasic) proliferation (red squares). 2 O 2 Co-expression of the scavenger catalase (CAT, green triangles) or mitochondria-targeted catalase (mCAT, blue inverted triangles) rescued the proliferation of cells lacking PRX3 expression.

[0014] Figure 2C Depicted are the weights (grams) of residual tumors excised from mice bearing MM xenografts in the peritoneal cavity following four weeks of treatment with 20 mg / ml thiostrepton composition twice weekly. **p<0.01

[0015] Figure 3A Depicted is a malignant pleural effusion (MPE) collected from a patient with metastatic disease.

[0016] Figure 3B Adherent tumor spheroids grown in MPE supernatant are depicted.

[0017] Figure 3CNon-adherent immune cells grown in MPE supernatant are depicted.

[0018] Figure 3D Depicted are the relative PRX3 inactivation by thiostrepton in tumor (adherent) and immune (non-adherent) cells.

[0019] Figure 3E This indicates that MPE-derived tumor cells are as sensitive to thiostrepton as established MM cell lines.

[0020] Figure 4A Drainage of pleural effusion to the trunk is depicted.

[0021] Figure 4B Depicted is the administration of thiostrepton using an indwelling intraperitoneal catheter (IPC).

[0022] Figure 4C Securing the IPC to the patient is depicted.

[0023] Figure 5 A summary of the thiostrepton dose escalation study is depicted.

[0024] Figure 6 Depicted is an exemplary Phase 1 dose escalation trial for treating cancer with the compounds and / or compositions of the present disclosure.

[0025] Figure 7 An exemplary Phase 1 dose expansion trial at RP2D is depicted, and TS represents a compound or composition of the present disclosure.

[0026] Figure 8 is a Swimmer plot of three exemplary patient results from the Phase 1 / 2 MITOPE trial. BOR indicates best overall response, PR indicates partial disease reduction, and SD indicates stable disease.

[0027] Fig. 9 The treatment profile of Patient 1 is shown.

[0028] Fig.10 is a graph showing the decrease in pleural effusion drainage volume over time for Patient 1.

[0029] Figures 11A-11C Shown are cross-sectional CT scans of the CP nodule of patient 1 before treatment (A), 6 weeks after treatment (B), and 12 weeks after treatment (C). The scans confirmed a partial response to treatment.

[0030] Figure 12A-12C Shown are cross-sectional CT scans of the right mediastinal pleural nodule of Patient 1 before treatment (A), 6 weeks after treatment (B), and 12 weeks after treatment (C). The scans confirmed a partial response to treatment.

[0031] Fig.13 The treatment profile of Patient 2 is shown.

[0032] Figure 14A-Figure 14C Shown are cross-sectional CT scans of the right pleural nodule of Patient 2 before treatment (A), 6 weeks after treatment (B), and 12 weeks after treatment (C). The scans confirmed stable disease.

[0033] Figure 15A-Figure 15C Cross-sectional CT scans of GB fundus lesions before treatment (A), 6 weeks after treatment (B), and 12 weeks after treatment (C) are shown for Patient 2. The scans confirmed stable disease.

[0034] Fig.16 is a graph showing the decrease in pleural effusion drainage volume over time for Patient 2.

[0035] Fig.17 The treatment profile of Patient 3 is shown.

[0036] Fig.18 is a graph showing the decrease in pleural effusion drainage volume over time in Patient 3.

[0037] Figure 19A-Figure 19C Cross-sectional CT scans of the tumor of Patient 3 before treatment (A), 6 weeks after treatment (B), and 12 weeks after treatment (C) are shown. The scans demonstrated stable disease up to 12 weeks after treatment.

[0038] Fig. 20 Shown is a cross-sectional CT scan of new liver metastases in Patient 3, indicating progressive disease at the week 12 scan.

[0039] Fig.21 Shown is a CT scan of new osteolytic bone metastases in Patient 3, indicating disease progression at the Week 12 scan. DETAILED DESCRIPTION

[0040] Tumor cells generate high levels of reactive oxygen species (ROS) and therefore exhibit increased expression and activity of key ROS scavenging pathways, including the mitochondrial peroxide scavenger enzyme peroxireductase 3 (PRX3). PRX3 is a peroxidase that is responsible for metabolizing approximately 90% of mitochondrial ROS, primarily H 2 O 2 . PRX3 transcription levels are upregulated in approximately 50% of cancers compared to normal tissues (data from the GEPIA2 database). Gene knockdown of PRX3 in human tumor cells resulted in sensitivity to apoptosis. Mitochondria in malignant mesothelioma (MM) cells are structurally and functionally altered, leading to metabolic dysfunction that supports tumor growth and could serve as a therapeutic target (see Figure 1A , Figure 1B ).

[0041] Thiostrepton is a covalent inhibitor that inactivates PRX3 peroxidase activity by direct adducting to the active site cysteine ​​residue, thereby inducing oxidative stress to levels incompatible with tumor cell survival. 50 exist Figure 2A Knockdown of PRX3 by siRNA significantly reduced MM (HM cell line-pleural biphasic) proliferation, as shown in Figure 2B As shown. 2 O 2 Co-expression of the scavenger catalase rescued the proliferation of cells lacking PRX3 expression. After treatment with 20 mg / ml thiostrepton composition twice a week for four weeks, the weight of the residual tumors excised from mice bearing MM xenografts in the peritoneal cavity was significantly less than that of the control ( Figure 2C ), thereby demonstrating a preclinical proof of principle for the biological activity of thiostrepton.

[0042] Small amounts of pleural fluid in the pleural cavity are physiologically normal. Mesothelioma and metastatic disease in the lungs commonly result in excessive fluid accumulation (approximately 15% of cancers). Malignant pleural effusions (MPE) are routinely drained using intrapleural catheters. Thiostrepton retains activity in patient-derived MPE. MPE contain both tumor and immune cells and allow for the evaluation of the efficacy of thiostrepton in this disease, where both cell types are inhibited by thiostrepton. Specifically, adherent tumor spheroids and non-adherent immune cells can be grown in culture medium containing MPE supernatant. ( Figure 3A , Figure 3B , Figure 3C In both tumor (adherent) and immune (non-adherent) cells, PRX3 was inactivated by thiostrepton ( Figure 3D ). In addition, MPE-derived tumor cells were equally sensitive to thiostrepton as established MM cell lines, such as Figure 3E shown.

[0043] In certain aspects, provided herein are methods of treating cancer comprising administering thiostrepton to a subject in need thereof once per week in a single dose of about 50 mg to about 500 mg, thereby treating the cancer.

[0044] In certain embodiments, a single dose is about 90 mg to about 450 mg. In some such embodiments, a single dose is selected from about 90 mg, about 120 mg, about 180 mg, about 270 mg, about 360 mg, and about 450 mg.

[0045] In some embodiments, a single dose is administered to a subject once a week for at least 3 weeks. In certain embodiments, a single dose in one week is the same as a single dose administered in a different week. In various embodiments, a first single dose is administered for 3 weeks, followed by a second single dose for the next 3 weeks, wherein the first single dose and the second single dose are different. In certain such embodiments, the second single dose is greater than the first single dose. In other embodiments, the second single dose is less than the first single dose.

[0046] In certain embodiments, thiostrepton is present in a pharmaceutical composition further comprising at least one pharmaceutically acceptable carrier.

[0047] In some embodiments, thiostrepton is administered topically. In certain embodiments, thiostrepton is administered intraperitoneally, intrapleurally, subcutaneously, or intratumorally.

[0048] In certain embodiments, thiostrepton is administered via a catheter, tube, or needle. In some embodiments, thiostrepton is administered via an indwelling intraperitoneal catheter (IPC).

[0049] In some embodiments, the cancer is selected from lung cancer, breast cancer, prostate cancer, melanoma, esophageal cancer, leukemia, cervical cancer, liver cancer, colon cancer, gastric cancer, colorectal cancer, glioblastoma, head and neck cancer, pancreatic cancer, mesothelioma and ovarian cancer. In further embodiments, the cancer is malignant mesothelioma. In further embodiments, the cancer is malignant pleural mesothelioma. In yet further embodiments, the cancer is malignant epithelioid pleural mesothelioma. In other embodiments, the cancer is malignant peritoneal mesothelioma. In certain embodiments, the cancer is lung cancer. In certain such embodiments, the lung cancer is adenocarcinoma. In some embodiments, the cancer is metastatic.

[0050] In certain embodiments, the subject experiences disease stabilization for at least 12 weeks after administering the first dose of thiostrepton. In some such embodiments, the subject experiences disease stabilization for at least about 12 weeks to about 52 weeks after administering the first dose of thiostrepton. In some embodiments, the subject experiences disease stabilization for at least 18 weeks after administering the first dose of thiostrepton. In some such embodiments, the subject experiences disease stabilization for at least about 18 weeks to about 52 weeks after administering the first dose of thiostrepton. In certain embodiments, the subject experiences disease stabilization for at least 24 weeks after administering the first dose of thiostrepton. In some such embodiments, the subject experiences disease stabilization for at least about 24 weeks to about 52 weeks after administering the first dose of thiostrepton.

[0051] In some embodiments, the subject experiences a progression-free survival period of at least 12 weeks after administering the first dose of thiostrepton. In some such embodiments, the subject experiences a progression-free survival period of at least about 12 weeks to about 52 weeks after administering the first dose of thiostrepton. In some embodiments, the subject experiences a progression-free survival period of at least 18 weeks after administering the first dose of thiostrepton. In certain such embodiments, the subject experiences a progression-free survival period of at least about 18 weeks to about 52 weeks after administering the first dose of thiostrepton. In certain embodiments, the subject experiences a progression-free survival period of at least 24 weeks after administering the first dose of thiostrepton. In some such embodiments, the subject experiences a progression-free survival period of at least about 24 weeks to about 52 weeks after administering the first dose of thiostrepton.

[0052] In certain embodiments, the subject experiences a partial response for at least 12 weeks after administering the first dose of thiostrepton. In some such embodiments, the subject experiences a partial response for at least about 12 weeks to about 52 weeks after administering the first dose of thiostrepton. In some embodiments, the subject experiences a partial response for at least 18 weeks after administering the first dose of thiostrepton. In some such embodiments, the subject experiences a partial response for at least about 18 weeks to about 52 weeks after administering the first dose of thiostrepton. In certain embodiments, the subject experiences a partial response for at least 24 weeks after administering the first dose of thiostrepton. In some such embodiments, the subject experiences a partial response for at least about 24 weeks to about 52 weeks after administering the first dose of thiostrepton.

[0053] In certain embodiments, a partial response comprises a reduction in tumor size by at least 10% relative to the size of the tumor prior to administration of thiostrepton. In further embodiments, a partial response comprises a reduction in tumor size by at least 30% relative to the size of the tumor prior to administration of thiostrepton. In still further embodiments, a partial response comprises a reduction in tumor size by at least 60% relative to the size of the tumor prior to administration of thiostrepton.

[0054] In certain embodiments, the subject experiences pleural effusion. In some such embodiments, 10 weeks after the first dose of thiostrepton is administered, the volume of the pleural effusion is reduced relative to the volume of the pleural effusion before the administration of thiostrepton. In certain embodiments, 5 weeks after the first dose of thiostrepton is administered, the volume of the pleural effusion is reduced relative to the volume of the pleural effusion before the administration of thiostrepton. In some embodiments, 1 week after the first dose of thiostrepton is administered, the volume of the pleural effusion is reduced relative to the volume of the pleural effusion before the administration of thiostrepton.

[0055] In certain embodiments, the volume of the pleural effusion is about 30% less than the volume of the pleural effusion before the administration of thiostrepton. In some embodiments, the volume of the pleural effusion is about 60% less than the volume of the pleural effusion before the administration of thiostrepton. In certain embodiments, the volume of the pleural effusion is about 90% less than the volume of the pleural effusion before the administration of thiostrepton.

[0056] In certain aspects, provided herein are pharmaceutical compositions comprising thiostrepton and at least one pharmaceutically acceptable carrier, wherein the pharmaceutical composition comprises about 50 mg to about 500 mg of thiostrepton.

[0057] In certain embodiments, the composition comprises about 90 mg to about 450 mg of thiostrepton. In some embodiments, the composition comprises about 90 mg, about 180 mg, about 270 mg, about 360 mg, or about 450 mg of thiostrepton.

[0058] The compositions and methods described herein can be used to treat individuals in need. The term "pharmaceutical composition" means a composition comprising thiostrepton and at least one pharmaceutically acceptable carrier. The terms "active compound" and "active ingredient" refer to thiostrepton. In certain embodiments, the individual is a mammal, such as a human or a non-human mammal. In certain preferred embodiments, the subject or mammal is a human. When applied to an animal such as a human, the composition or compound is preferably administered as a pharmaceutical composition comprising, for example, an active compound as described herein and a pharmaceutically acceptable carrier. Pharmaceutically acceptable carriers are well known in the art and, as non-limiting examples, include aqueous solutions such as water or physiologically buffered saline or other solvents or vehicles such as glycols, glycerol, oils such as olive oil or injectable organic esters. In preferred embodiments, when such pharmaceutical compositions are used for human administration, particularly for invasive routes of administration (i.e., routes of injection or implantation that circumvent transport or diffusion through an epithelial barrier), the aqueous solution is pyrogen-free or substantially pyrogen-free. Excipients can be selected, for example, to achieve delayed release of the agent or selectively target one or more cells, tissues or organs. The pharmaceutical composition can be in dosage unit form, such as tablets, capsules (including dispersible capsules and gelatin capsules), granules, lyophilized agents for reconstitution, powders, solutions, syrups, suppositories, injections, etc. The composition can also be present in a transdermal delivery system, such as a skin patch. The composition can also be present in a solution suitable for topical application, such as a lotion, cream or ointment.

[0059] A pharmaceutically acceptable carrier may contain a physiologically acceptable agent, which, for example, acts to stabilize an active compound (such as an active compound described herein), increase its solubility, or increase its absorption. Such physiologically acceptable agents include, for example, carbohydrates such as glucose, sucrose, or dextran; antioxidants such as ascorbic acid or glutathione; chelating agents; low molecular weight proteins or other stabilizers or excipients. The selection of a pharmaceutically acceptable carrier (including a physiologically acceptable agent) depends, for example, on the route of administration of the composition. The formulation or pharmaceutical composition may be a self-emulsifying drug delivery system or a self-microemulsifying drug delivery system. The pharmaceutical composition (preparation) may also be a liposome or other polymer matrix, into which a compound such as described herein may be incorporated. For example, liposomes comprising phospholipids or other lipids are non-toxic, physiologically acceptable, and metabolizable carriers that are relatively simple to prepare and administer.

[0060] The phrase "pharmaceutically acceptable" is used herein to refer to those compounds, materials, compositions and / or dosage forms which are, within the scope of sound medical judgment, suitable for use in contact with the tissues of humans and animals without excessive toxicity, irritation, allergic response, or other problems or complications, commensurate with a reasonable benefit / risk ratio.

[0061] As used herein, the phrase "pharmaceutically acceptable carrier" means a pharmaceutically acceptable material, composition, or vehicle, such as a liquid or solid filler, diluent, excipient, solvent, or encapsulating material. Each carrier must be "acceptable" in the sense of being compatible with the other ingredients of the formulation and not injurious to the patient. Some examples of materials that can be used as pharmaceutically acceptable carriers include: (1) sugars such as lactose, glucose and sucrose; (2) starches such as corn starch and potato starch; (3) cellulose and derivatives such as sodium carboxymethylcellulose, ethylcellulose and cellulose acetate; (4) tragacanth powder; (5) malt; (6) gelatin; (7) talc; (8) excipients such as cocoa butter and suppository waxes; (9) oils such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil and soybean oil; (10) glycols such as propylene glycol; (11) polyols such as glycerol, sorbitol, mannitol and polyethylene glycol; (12) esters such as ethyl oleate and ethyl laurate; (13) agar; (14) buffers such as magnesium hydroxide and aluminum hydroxide; (15) alginic acid; (16) pyrogen-free water; (17) isotonic saline; (18) Ringer's solution solution); (19) ethanol; (20) phosphate buffer solution; and (21) other non-toxic compatible substances used in pharmaceutical formulations.

[0062] The pharmaceutical composition (preparation) can be administered to a subject by any of a variety of routes of administration, including, for example, intraperitoneal; intrapleural; subcutaneous; intratumoral; oral (e.g., an infusion in water or non-aqueous solution or suspension for application to the tongue, tablets, capsules (including dispersible capsules and gelatin capsules), boluses, powders, granules, pastes); absorption through the oral mucosa (e.g., sublingual); transdermal (e.g., as a patch applied to the skin); and topical administration (e.g., as a cream, ointment or spray applied to the skin). The compound can also be formulated for inhalation. In some embodiments, the pharmaceutical composition is topically administered. In some embodiments, the pharmaceutical composition is administered by a catheter, tube or needle. In certain embodiments, the compound can simply be dissolved or suspended in sterile water. Details of appropriate routes of administration and compositions useful therefor may be found, for example, in U.S. Pat. Nos. 6,110,973, 5,763,493, 5,731,000, 5,541,231, 5,427,798, 5,358,970, and 4,172,896, and patents cited therein.

[0063] The formulation may conveniently be presented in unit dosage form and may be prepared by any method well known in the pharmaceutical art. The amount of active ingredient that may be combined with a carrier material to produce a single dosage form will vary depending on the host being treated, the particular mode of administration. The amount of active ingredient that may be combined with a carrier material to produce a single dosage form will generally be the amount of compound that produces a therapeutic effect.

[0064] The methods for preparing these formulations or compositions include the step of bringing into association the active compound (such as thiostrepton) with a carrier and optionally one or more accessory ingredients. In general, the formulations are prepared by uniformly and intimately bringing into association the compound described herein with liquid carriers, or finely divided solid carriers, or both, and then, if necessary, shaping the product.

[0065] Liquid dosage forms include pharmaceutically acceptable emulsions, lyophilized agents for reconstitution, microemulsions, solutions, suspensions, syrups and elixirs. In addition to the active ingredient, the liquid dosage form may contain an inert diluent commonly used in the art, such as, for example, water or other solvents, cyclodextrins and derivatives thereof, solubilizers and emulsifiers, such as ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butylene glycol, oils (particularly cottonseed oil, peanut oil, corn oil, germ oil, olive oil, castor oil and sesame oil), glycerol, tetrahydrofurfuryl alcohol, polyethylene glycol and fatty acid esters of sorbitan and mixtures thereof.

[0066] Besides inert diluents, the liquid composition may also include adjuvants such as wetting agents, emulsifying and suspending agents, sweetening, flavoring, coloring, perfuming and preservatives.

[0067] Suspensions, in addition to the active compounds, may contain suspending agents such as ethoxylated isostearyl alcohols, polyoxyethylene sorbitol and sorbitan esters, microcrystalline cellulose, aluminum metahydroxide, bentonite, agar-agar and tragacanth, and mixtures thereof.

[0068] As used herein, the phrases "parenteral administration" and "administered parenterally" mean modes of administration other than enteral administration and topical administration, usually by injection, and include, but are not limited to, intraperitoneal, intrapleural, subcutaneous, intratumoral, intravenous, intramuscular, intraarterial, intrathecal, intracapsular, intraorbital, intracardiac, intradermal, transtracheal, subcutaneous, intraarticular, subcapsular, subarachnoid, intraspinal, and intrasternal injection and infusion. In some embodiments, the pharmaceutical composition is administered intraperitoneally, intrapleurally, subcutaneously, or intratumorally. Pharmaceutical compositions suitable for parenteral administration comprise a combination of one or more active compounds with one or more pharmaceutically acceptable sterile isotonic aqueous or non-aqueous solutions, dispersions, suspensions, or emulsions, or sterile powders that can be reconstituted into sterile injectable solutions or dispersions immediately prior to use, which combination may contain antioxidants, buffers, bacteriostats, solutes that render the formulation isotonic with the blood of the intended recipient, or suspending or thickening agents.

[0069] Examples of suitable aqueous and nonaqueous carriers that can be used in the pharmaceutical compositions described herein include water, ethanol, polyols (such as glycerol, propylene glycol, polyethylene glycol, etc.), and suitable mixtures thereof, vegetable oils (such as olive oil), and injectable organic esters (such as ethyl oleate). Proper fluidity can be maintained, for example, by the use of coating materials (such as lecithin), by maintaining the required particle size in the case of dispersions, and by the use of surfactants.

[0070] These compositions may also contain adjuvants, such as preservatives, wetting agents, emulsifiers and dispersants. Various antibacterial and antifungal agents, such as parabens, chlorobutanol, phenol, sorbic acid, etc., may be included to ensure that the effects of microorganisms are prevented. It may also be desirable to include isotonic agents, such as sugar, sodium chloride, etc., in the composition. In addition, extended absorption of injectable drug forms may be achieved by including agents that delay absorption, such as aluminum monostearate and gelatin.

[0071] In some cases, in order to prolong the effect of the drug, it is desirable to slow down the injection, for example, the absorption of the drug after subcutaneous injection. This can be achieved by using a liquid suspension of a crystalline or amorphous material with poor water solubility. The absorption rate of the drug depends on its dissolution rate, which in turn can depend on crystal size and crystalline form. Alternatively, the delayed absorption of the drug form administered parenterally can be accomplished by dissolving or suspending the drug in an oil vehicle.

[0072] Injectable depot forms are prepared by forming a microencapsulated matrix of the subject compound in a biodegradable polymer such as polylactide-polyglycolide. Depending on the ratio of the drug to the polymer and the properties of the specific polymer used, the drug release rate can be controlled. Examples of other biodegradable polymers include poly(orthoesters) and poly(anhydrides). Depot injection formulations are also prepared by embedding the drug in liposomes or microemulsions compatible with body tissues.

[0073] For use in the methods described herein, the active compound may be provided per se or as a pharmaceutical composition containing, for example, 0.1% to 99.5% (more preferably 0.5% to 90%) active ingredient in combination with a pharmaceutically acceptable carrier.

[0074] The method of introduction can also be provided by a refillable or biodegradable device. Regarding the controlled delivery of drugs (including protein biopharmaceuticals), various slow-release polymer devices have been developed and tested in vivo in recent years. A variety of biocompatible polymers (including hydrogels), including both biodegradable and non-degradable polymers, can be used to form implants to continuously release compounds at specific target sites.

[0075] Actual dosage levels of the active ingredients in the pharmaceutical compositions may be varied so as to obtain an amount of the active ingredient which is effective to achieve the desired therapeutic response for a particular patient, composition, and mode of administration, without being toxic to the patient.

[0076] The selected dosage level will depend upon a variety of factors including the activity of the specific compound or combination of compounds employed, or the esters, salts or amides thereof, the route of administration, the time of administration, the rate of excretion of the specific compound employed, the duration of the treatment, other drugs, compounds and / or materials used in combination with the specific compound employed, the age, sex, weight, condition, general health and prior medical history of the patient being treated, and like factors well known in the medical arts.

[0077] A physician or veterinarian with ordinary skills in the art can easily determine and prescribe a therapeutically effective amount of the desired pharmaceutical composition. For example, a physician or veterinarian can start the dosage of the pharmaceutical composition or compound at a level lower than the level required to achieve the desired therapeutic effect and gradually increase the dosage until the desired effect is achieved. "Therapeutically effective amount" means the concentration of the compound sufficient to cause the desired therapeutic effect. It is generally understood that the effective amount of the compound will vary according to the subject's weight, sex, age and medical history. Other factors affecting the effective amount may include, but are not limited to, the severity of the patient's illness, the condition being treated, the stability of the compound, and, if desired, another type of therapeutic agent administered with the compound described herein. A larger total dose can be delivered by multiple administrations of the agent. Methods for determining efficacy and dosage are known to those skilled in the art (Isselbacher et al. (1996) Harrison's Principles of Internal Medicine 13th edition, 1814-1882, incorporated herein by reference).

[0078] In general, a suitable dose of the active compound used in the compositions and methods described herein will be that amount of the compound which is the lowest dose effective to produce a therapeutic effect. Such an effective dose will generally depend on the factors described above.

[0079] If desired, the effective dose of the active compound can be administered as one, two, three, four, five, six or more sub-doses, separated by appropriate intervals, optionally in unit dosage form. In some embodiments, thiostrepton is administered once a week. In certain embodiments, thiostrepton is administered for two, three, four, five or six consecutive weeks. In some embodiments, thiostrepton is administered every other week, once every three weeks or once every four weeks at a rate of once or twice a week.

[0080] Patients receiving such treatment are any animal in need thereof, including primates, particularly humans; and other mammals, such as horses, cattle, pigs, sheep, cats, and dogs; poultry; and pets in general.

[0081] In certain embodiments, the compounds described herein may be used alone or in combination with another type of therapeutic agent.

[0082] The present disclosure includes the use of pharmaceutically acceptable salts of compounds described herein in compositions and methods described herein. In certain embodiments, the salts considered include but are not limited to alkyl, dialkyl, trialkyl or tetraalkyl ammonium salts. In certain embodiments, the salts considered include but are not limited to L-arginine, benethamine, benzathine, betaine, calcium hydroxide, choline, deanol, diethanolamine, diethylamine, 2-(diethylamino)ethanol, ethanolamine, ethylenediamine, N-methylglucosamine, hydrabamine, 1H-imidazole, lithium, L-lysine, magnesium, 4-(2-hydroxyethyl)morpholine, piperazine, potassium, 1-(2-hydroxyethyl)pyrrolidine, sodium, triethanolamine, tromethamine and zinc salts. In certain embodiments, the salts considered include but are not limited to Na, Ca, K, Mg, Zn or other metal salts. In certain embodiments, contemplated salts include, but are not limited to, 1-hydroxy-2-naphthoic acid, 2,2-dichloroacetic acid, 2-hydroxyethanesulfonic acid, 2-oxoglutaric acid, 4-acetamidobenzoic acid, 4-aminosalicylic acid, acetic acid, adipic acid, 1-ascorbic acid, l-aspartic acid, benzenesulfonic acid, benzoic acid, (+)-camphoric acid, (+)-camphor-10-sulfonic acid, capric acid / decanoic acid, caproic acid / hexanoic acid, caprylic acid / octanoic acid, carbonic acid, cinnamic acid, citric acid, cyclamic acid, acid), dodecyl sulfuric acid, ethane-1,2-disulfonic acid, ethanesulfonic acid, formic acid, fumaric acid, galactaric acid, gentisic acid, d-glucoheptonic acid, d-gluconic acid, d-glucuronic acid, glutamic acid, glutaric acid, glycerophosphoric acid, glycolic acid, hippuric acid, hydrobromic acid, hydrochloric acid, isobutyric acid, lactic acid, lactobionic acid, lauric acid, maleic acid, 1-malic acid, malonic acid, phenylglycolic acid, methanesulfonic acid, naphthalene-1,5-disulfonic acid, naphthalene-2-sulfonic acid, niacin, nitric acid, oleic acid, oxalic acid, palmitic acid, pamoic acid, phosphoric acid, propionic acid, l-pyroglutamic acid, salicylic acid, sebacic acid, stearic acid, succinic acid, sulfuric acid, 1-tartaric acid, thiocyanic acid, p-toluenesulfonic acid, trifluoroacetic acid, and undecylenate.

[0083] Pharmaceutically acceptable acid addition salts may also exist in the form of various solvates, such as solvates with water, methanol, ethanol, dimethylformamide, etc. Mixtures of such solvates may also be prepared. The source of such solvates may be from the solvent of crystallization, inherent in the solvent of preparation or crystallization, or insoluble in such solvents.

[0084] Wetting agents, emulsifiers and lubricants, such as sodium lauryl sulfate and magnesium stearate, as well as coloring agents, release agents, coating agents, sweetening, flavoring and perfuming agents, preservatives and antioxidants can also be present in the compositions.

[0085] Examples of pharmaceutically acceptable antioxidants include: (1) water-soluble antioxidants, such as ascorbic acid, cysteine ​​hydrochloride, sodium bisulfate, sodium metabisulfite, sodium sulfite, etc.; (2) oil-soluble antioxidants, such as ascorbyl palmitate, butylated hydroxyanisole (BHA), butylated hydroxytoluene (BHT), lecithin, propyl gallate, α-tocopherol, etc.; and (3) metal chelators, such as citric acid, ethylenediaminetetraacetic acid (EDTA), sorbitol, tartaric acid, phosphoric acid, etc.

[0086] definition

[0087] Unless otherwise defined herein, scientific and technical terms used in this application shall have the meanings commonly understood by those of ordinary skill in the art. In general, the nomenclature and techniques used in conjunction with chemistry, cell and tissue culture, molecular biology, cell and cancer biology, neurobiology, neurochemistry, virology, immunology, microbiology, pharmacology, genetics, and protein and nucleic acid chemistry described herein are those well-known and commonly used in the art.

[0088] The methods and techniques of the present disclosure are generally performed according to conventional methods well known in the art and as described in various general and more specific references that are cited and discussed throughout the present specification unless otherwise indicated.

[0089] All of the above and any other publications, patents, and published patent applications mentioned in this patent application are expressly incorporated herein by reference. In the event of a conflict, the present specification (including its specific definitions) will control.

[0090] The term "agent" as used herein refers to a compound (such as an organic or inorganic compound, a mixture of compounds), a biomacromolecule (such as a nucleic acid, an antibody, including parts thereof and humanized, chimeric and human antibodies and monoclonal antibodies, a protein or a part thereof, such as a peptide, a lipid, a carbohydrate) or an extract made from a biological material such as a bacterium, a plant, a fungus or an animal (particularly a mammal) cell or tissue. Agents include, for example, agents with known structures and agents with unknown structures.

[0091] "Patient", "subject" or "individual" are used interchangeably and refer to humans or non-human animals. These terms include mammals, such as humans, primates, livestock animals (including cattle, pigs, etc.), companion animals (e.g., dogs, cats, etc.) and rodents (e.g., mice and rats). The subject may be male or female. In some embodiments, the subject is older than 18 years old. In certain embodiments, the subject's ECOG (Eastern Cooperative Oncology Group) score is preferably 0-1. In certain embodiments, the patient may have a histological diagnosis of MPE caused by a non-mesothelioma solid tumor or mesothelioma. In some embodiments, the patient has received at least one previous standard of care treatment regimen, with a record of progression and no approved alternatives available. In some embodiments, all acute reversible toxic effects of the patient's previous therapy have subsided to ≤ grade 1. In certain embodiments, the patient has a paraffin block of his / her most recent biopsy. In some embodiments, the patient has adequate organ function before administering thiostrepton, as defined by laboratory values. In some embodiments, the subject is postmenopausal, surgically sterilized, or using effective contraception.

[0092] In some embodiments, the patient has not received existing systemic anticancer or radiotherapy before the administration of thiostrepton. In some embodiments, the patient has not undergone surgery within 3 weeks or 5 half-lives before the administration of thiostrepton. In some embodiments, the patient has not been treated with a research product / device within 30 days before the administration of thiostrepton. In certain embodiments, the patient has not previously suffered from other malignancies except for the cancer to be treated before the administration of thiostrepton. In certain embodiments, the patient does not have a tumor or loculation that causes incomplete or ineffective intrapleural administration. In certain embodiments, the patient has no known hypersensitivity to thiostrepton or pharmaceutical composition excipients. In some embodiments, the patient does not have any surgery or medical condition that may interfere with thiostrepton treatment. In some embodiments, the patient has no human immunodeficiency virus (HIV) or hepatitis B in the absence of a sustained virological response; or active infection with hepatitis C. In certain embodiments, the patient is not pregnant or breastfeeding. In some embodiments, the patient has no symptomatic or unstable CNS tumors or metastases or carcinomatous meningitis. In some embodiments, the patient has not used systemic corticosteroids within 15 days prior to the administration of thiostrepton, or other immunosuppressive drugs within 3 weeks prior to the administration of thiostrepton.

[0093] "Treatment" of a disease or patient refers to taking steps to obtain beneficial or desired results, including clinical results. As used herein and as well understood in the art, "treatment" is a means for obtaining beneficial or desired results (including clinical results). Beneficial or desired clinical results may include, but are not limited to, the mitigation or improvement of one or more symptoms or conditions, the reduction of the extent of the disease, the stabilization of the disease state (i.e., no deterioration), the prevention of disease spread, the delay or slowing of the disease process, the improvement or alleviation of the disease state, and relief (whether partial relief or total relief), whether detectable or undetectable. "Treatment" may also mean prolonged survival compared to the survival expected when not receiving treatment.

[0094] The term "prevention" is art-recognized and when used with respect to a condition such as a local recurrence (e.g., pain), a disease such as cancer, a symptom such as heart failure, or any other medical condition, is well known in the art and includes administering a composition that reduces the frequency of symptoms of the medical condition in a subject, or delays the onset thereof, relative to a subject not receiving the composition. Thus, prevention of cancer includes, for example, reducing the number of detectable cancerous growths in a patient population receiving prophylactic treatment relative to an untreated control population, and / or delaying the appearance of detectable cancerous growths in a treated population relative to an untreated control population, for example, by a statistically and / or clinically significant amount.

[0095] "Administering / administration of" a substance, compound or agent to a subject can be performed using one of the various methods known to those skilled in the art. For example, a compound or agent can be administered intraperitoneally, intrapleurally, subcutaneously, intratumorally, intravenously, intraarterially, intradermally, intramuscularly, subcutaneously, ocularly, sublingually, orally (by ingestion), intranasally (by inhalation), intraspinal, intracerebrally, and transdermally (by absorption, for example, through a skin tube). The compound or agent can also be suitably introduced by a rechargeable or biodegradable polymer device or other device, for example, a patch and a pump, or a formulation that provides extended, slow or controlled release of the compound or agent. Administration can also be performed, for example, once, multiple times, and / or over one or more extended periods of time.

[0096] In certain embodiments, administration of thiostrepton or any pharmaceutical composition comprising thiostrepton disclosed herein can be performed using an indwelling intraperitoneal catheter (IPC). In some embodiments, administration occurs once a week or twice a week, preferably once a week. In some embodiments, the method further comprises, for example, removing fluid from the pleural effusion prior to administering thiostrepton. Figure 4A In certain embodiments, after administration of thiostrepton ( Figure 4B ), IPC was fixed until the next dosing time point ( Figure 4C ).

[0097] In certain embodiments, the single dose amount of thiostrepton ranges from about 50 mg to about 500 mg. In some embodiments, the single dose amount of thiostrepton ranges from about 90 mg to about 450 mg. In preferred embodiments, the single dose is selected from about 90 mg, about 180 mg, about 270 mg, about 360 mg, and about 450 mg. In some embodiments, a single dose is administered to a subject once a week, for example, for at least 3 weeks.

[0098] In some embodiments, the single dose of thiostrepton may be increased once every three-week period, for example, such that the patient is dosed with 90 mg once a week during weeks 1-3, then optionally with 180 mg once a week during weeks 4-6, then optionally with 270 mg once a week during weeks 7-9, then optionally with 360 mg once a week during weeks 10-12, and finally optionally with 450 mg once a week during weeks 13-15. Each patient may complete any or all of the 3-week courses ( Figure 5 ). In some embodiments, the dosing regimen may be paused, stopped, or the patient may be switched to a lower dose, for example, upon the occurrence of toxicity or adverse events.

[0099] In some embodiments, the method further comprises obtaining a tumor biopsy from the patient prior to administering thiostrepton. In some embodiments, the method further comprises obtaining a tumor biopsy after administering a third dose of thiostrepton.

[0100] In other embodiments, the pharmaceutical composition comprises about 5 mg thiostrepton / mL to about 50 mg thiostrepton / mL. In some embodiments, the pharmaceutical composition comprises about 10 mg thiostrepton / mL, about 20 mg thiostrepton / mL, about 30 mg thiostrepton / mL, about 40 mg thiostrepton / mL and about 50 mg thiostrepton / mL. In a preferred embodiment, the pharmaceutical composition comprises 20 mg thiostrepton / mL.

[0101] The appropriate method of administering a substance, compound or agent to a subject will also depend on, for example, the age and / or physical condition of the subject and the chemical and biological properties (e.g., solubility, digestibility, bioavailability, stability and toxicity) of the compound or agent. In some embodiments, the compound or agent is orally administered to the subject, for example, by ingestion. In some embodiments, the orally administered compound or agent is in an extended release or slow release formulation, or is administered using a device for such slow or extended release.

[0102] As used herein, the phrase "combined administration" refers to any form of administration of two or more different therapeutic agents, such that a second dose is administered when the previously administered therapeutic agent is still effective in the body (e.g., when at least 5% of the drug product can be systematically detected by industry-accepted methods, or when the two agents are simultaneously effective in the patient, which may include a synergistic effect of the two agents). For example, different therapeutic compounds may be administered simultaneously or sequentially in the same formulation or in separate formulations. In certain embodiments, different therapeutic compounds may be administered within one hour, 12 hours, 24 hours, 36 hours, 48 ​​hours, 72 hours, or a week of each other. Therefore, individuals receiving such treatment may benefit from the combined effects of different therapeutic agents.

[0103] In some embodiments, the amount of the first single dose in one week is the same as the amount of the single dose administered in a different week. In certain embodiments, the first single dose is administered for 3 weeks, followed by the second single dose for the next 3 weeks, wherein the first single dose and the second single dose are different. In some embodiments, the second single dose is larger than the first single dose. In other embodiments, the second single dose is smaller than the first single dose.

[0104] A "therapeutically effective amount" or "therapeutically effective dose" of a compound or other agent described herein is an amount of the drug or agent that will have the intended therapeutic effect when administered to a subject. The full therapeutic effect does not necessarily occur by administering one dose of such a drug or agent, but may only occur after administering a series of doses (multiple consecutive doses). Thus, a therapeutically effective amount may be administered in one or more administrations. The precise effective amount required for a subject will depend, for example, on the subject's size, health, and age, as well as the nature and extent of the disorder (such as cancer) being treated.

[0105] As used herein, the terms "optional" or "optionally" mean that the subsequently described event or circumstance may or may not occur, and that the description includes instances where the event or circumstance occurs and instances where it does not.

[0106] As used herein, the term "modulate" includes inhibiting or suppressing a function or activity (such as cell proliferation) as well as enhancing a function or activity.

[0107] The phrase "pharmaceutically acceptable" is recognized in the art. In certain embodiments, the term includes compositions, excipients, adjuvants, polymers and other materials and / or dosage forms that are suitable for use in contact with the tissues of humans and animals without excessive toxicity, irritation, allergic response or other problems or complications, and commensurate with a reasonable benefit / risk ratio, within the scope of sound medical judgment.

[0108] In certain embodiments, disclosed herein is a method for treating cancer (e.g., solid tumors or hematological cancers), comprising administering to a subject in need thereof a therapeutically effective amount of any compound described herein or a composition of the compound. In certain embodiments, cancer (solid tumor or hematological tumor) is selected from lung cancer, breast cancer, prostate cancer, melanoma, esophageal cancer, leukemia, cervical cancer, liver cancer, colon cancer, gastric cancer, colorectal cancer, glioblastoma, head and neck cancer, pancreatic cancer, mesothelioma and ovarian cancer. In certain embodiments, cancer is selected from mesothelioma, lung cancer, ovarian cancer and breast cancer. In some embodiments, cancer is malignant mesothelioma.

[0109] Example

[0110] Exemplary Phase 1 / 2 Dose Escalation and Expansion Study

[0111] A Phase 1 / 2 dose escalation and expansion study was conducted to determine the safety, tolerability, and recommended Phase 2 dose of an exemplary thiostrepton composition in patients with malignant pleural effusions due to advanced / metastatic solid tumors, including mesothelioma. Preliminary results for three patients (Patient 1, Patient 2, and Patient 3) are provided herein. Figure 6-Figure 20 An overview of the trial, treatment details, and exemplary preliminary data for these three patients is provided.

[0112] The terms "partial response" ("PR"), "stable disease" ("SD"), and "best overall response" ("BOR") are used according to the definitions and criteria set forth in the RECIST guidelines, version 1.1 (Eur. J. Cancer. 2009, 45, 228-247).

[0113] Patient 1:

[0114] 84-year-old male, ECOG 1

[0115] Background medical history: hypertension, hypercholesterolemia, benign prostatic hyperplasia

[0116] Diagnosed with malignant epithelioid pleural mesothelioma in December 2020

[0117] Previous first-line (1L) therapy: carboplatin + pemetrexed BOR = stable disease

[0118] Patients progressed after 1L therapy

[0119] Treatment with thiostrepton at a dose of 90 mg once a week

[0120] like Fig. 9 As shown in the graph in , patient 1 showed a partial response (approximately 59% reduction in tumor size) 18 weeks after initial thiostrepton treatment and had stable disease at week 24. Fig.10It was shown that patient 1 experienced a significant reduction in pleural effusion through 30 weeks of thiostrepton treatment. Figures 11A-11C Patient 1 before treatment ( Fig.11A ), 6 weeks after treatment ( Fig. 11B ) and 12 weeks after treatment ( Fig. 11C ) CT scan.

[0121] Patient 2:

[0122] 75-year-old female, ECOG 1

[0123] On April 12, 2019, he was diagnosed with malignant epithelioid pleural mesothelioma with subdiaphragmatic disease.

[0124] Background medical history: hypertension, bronchiectasis complicated by TB 25 years ago.

[0125] 1 st L: Chemotherapy carboplatin + pemetrexed BOR = stable disease

[0126] 2 nd L: Vinorelbine; BOR = PD

[0127] ·3 rd L: Nivolumab; BOR = SD

[0128] ·4 rd L: gemcitabine; BOR = PD

[0129] 5 th L: Pem / Carbo (reattack); BOR = PR

[0130] Patient progressed after Carb / Pem rechallenge

[0131] Treatment with 90 mg thiostrepton once a week

[0132] like Fig.13 As shown, Patient 2 showed stable disease through at least 18 weeks of treatment. Fig.16 Patient 2 experienced a reduction in the amount of pleural effusion drained through 18 weeks of treatment. Figure 14A-Figure 14C and Figure 15A-Figure 15C Patient 2 before treatment ( Fig.14A , Fig.15A ), 6 weeks after treatment ( Fig. 14B and Fig. 15B ) and 12 weeks after treatment ( Fig. 14C and Fig. 15C ) CT scan.

[0133] Patient 3:

[0134] 74-year-old male, ECOG 1

[0135] Diagnosed as T4N2M1a lung adenocarcinoma on July 17, 2021

[0136] Background medical history: glaucoma, atrial flutter - treated with ablation.

[0137] 1 st Line: Carboplatin + Pemetrexed + Pembro; BOR = stable disease

[0138] The patient progressed on 25 / 02 / 23

[0139] Significant pleural effusion-MPE

[0140] · IPC inserted on 03 / 09 / 23 - draining large hemorrhagic pleural effusions, >4 liters, weekly.

[0141] · Trial treatment started on 17MAR23

[0142] Treatment with 90 mg thiostrepton once a week

[0143] like Fig.17 As shown, patient 3 showed stable disease through at least 6 weeks of treatment, with progressive disease observed on the week 12 scan. Fig.18 Patient 3 experienced a reduction in the amount of pleural effusion drained through 12 weeks of treatment. Figure 19A-Figure 19C Patient 3 before treatment ( Fig.19A ), 6 weeks after treatment ( Fig.19B ) and 12 weeks after treatment ( Fig.19C ) CT scan. Fig. 20 and Fig.21 This is a CT scan of patient 3 at 12 weeks of treatment.

[0144] Incorporated by Reference

[0145] All publications and patents mentioned herein are hereby incorporated by reference in their entirety to the same extent as if each individual publication or patent was specifically and individually indicated to be incorporated by reference. In the event of a conflict, the present application, including any definitions herein, will control.

[0146] Equivalent solutions

[0147] The foregoing written description is considered sufficient to enable those skilled in the art to practice the present invention. The scope of the present invention is not limited by the examples provided, as the examples are only used to illustrate one aspect of the present invention alone, and other functionally equivalent embodiments are also within the scope of the present invention. In addition to the modifications shown and described herein, various modifications of the present invention will become apparent to those skilled in the art from the foregoing description and fall within the scope of the appended claims. The advantages and objects of the present invention are not necessarily encompassed in every embodiment of the present invention.

Claims

1. A method of treating cancer, the method comprising administering thiostrepton to a subject in need thereof once a week in a single dose of about 50 mg to about 500 mg, thereby treating the cancer.

2. The method of claim 1, wherein the single dose is from about 90 mg to about 450 mg.

3. The method of claim 2, wherein the single dose is selected from about 90 mg, about 120 mg, about 180 mg, about 270 mg, about 360 mg, and about 450 mg.

4. The method of any preceding claim, wherein the single dose is administered to the subject once a week for at least 3 weeks.

5. The method of claim 4, wherein the amount of the single dose within one week is the same as the amount of the single dose administered in a different week.

6. The method of claim 4, wherein a first single dose is administered for 3 weeks followed by a second single dose for the next 3 weeks, wherein the first single dose and the second single dose are different.

7. The method of claim 6, wherein the second single dose is greater than the first single dose.

8. The method of claim 6, wherein the second single dose is less than the first single dose.

9. The method of any one of claims 1 to 8, wherein the thiostrepton is present in a pharmaceutical composition further comprising at least one pharmaceutically acceptable carrier.

10. The method of any one of claims 1 to 9, wherein the thiostrepton is administered topically.

11. The method of any one of claims 1 to 10, wherein the thiostrepton is administered intraperitoneally, intrapleurally, subcutaneously, or intratumorally.

12. The method of any one of claims 1 to 11, wherein the thiostrepton is administered via a catheter, tube, or needle.

13. The method of any one of claims 1 to 10, wherein the thiostrepton is administered via an indwelling intraperitoneal catheter (IPC).

14. The method of any one of claims 1 to 10, wherein the cancer is selected from lung cancer, breast cancer, prostate cancer, melanoma, esophageal cancer, leukemia, cervical cancer, liver cancer, colon cancer, gastric cancer, colorectal cancer, glioblastoma, head and neck cancer, pancreatic cancer, mesothelioma, and ovarian cancer.

15. The method of any one of claims 1 to 10, wherein the cancer is malignant mesothelioma.

16. The method of claim 15, wherein the cancer is malignant pleural mesothelioma.

17. The method of claim 16, wherein the cancer is malignant epithelioid pleural mesothelioma.

18. The method of claim 16, wherein the cancer is malignant peritoneal mesothelioma.

19. The method of claim 14, wherein the cancer is lung cancer.

20. The method of claim 19, wherein the lung cancer is adenocarcinoma.

21. The method of any one of claims 14 to 20, wherein the cancer is metastatic.

22. The method of any preceding claim, wherein the subject experiences stable disease for at least 12 weeks following administration of the first dose of thiostrepton.

23. The method of any preceding claim, wherein the subject experiences stable disease for at least 18 weeks following administration of the first dose of thiostrepton.

24. The method of any one of claims 1 to 21, wherein the subject experiences stable disease for at least 24 weeks after administration of the first dose of thiostrepton.

25. The method of any preceding claim, wherein the subject experiences progression-free survival of at least 12 weeks after administration of the first dose of thiostrepton.

26. The method of any preceding claim, wherein the subject experiences progression-free survival of at least 18 weeks after administration of the first dose of thiostrepton.

27. The method of any one of claims 1 to 21, wherein the subject experiences progression-free survival of at least 24 weeks after administration of the first dose of thiostrepton.

28. The method of any one of claims 1 to 21, wherein the subject experiences a partial response for at least 12 weeks after administration of the first dose of thiostrepton.

29. The method of any one of claims 1 to 21, wherein the subject experiences a partial response for at least 18 weeks after administration of the first dose of thiostrepton.

30. The method of any one of claims 1 to 21, wherein the subject experiences a partial response for at least 24 weeks after administration of the first dose of thiostrepton.

31. The method of any one of claims 28 to 30, wherein the partial response comprises a decrease in tumor size of at least 10% relative to tumor size prior to administration of thiostrepton.

32. The method of any one of claims 28 to 31, wherein the partial response comprises a decrease in tumor size of at least 30% relative to tumor size prior to administration of thiostrepton.

33. The method of any one of claims 28 to 32, wherein the partial response comprises a reduction in tumor size of at least 60% relative to tumor size prior to administration of thiostrepton.

34. The method of any preceding claim, wherein the subject experiences a pleural effusion.

35. The method of claim 34, wherein 10 weeks after administration of the first dose of thiostrepton, the volume of the pleural effusion is reduced relative to the volume of the pleural effusion prior to administration of thiostrepton.

36. The method of claim 34, wherein 5 weeks after administration of the first dose of thiostrepton, the volume of the pleural effusion is reduced relative to the volume of the pleural effusion prior to administration of thiostrepton.

37. The method of claim 34, wherein one week after administration of the first dose of thiostrepton, the volume of the pleural effusion is reduced relative to the volume of the pleural effusion prior to administration of thiostrepton.

38. The method of any one of claims 35 to 37, wherein the volume of the pleural effusion is about 30% less than the volume of the pleural effusion prior to administration of thiostrepton.

39. The method of any one of claims 35 to 37, wherein the volume of the pleural effusion is about 60% less than the volume of the pleural effusion prior to administration of thiostrepton.

40. The method of any one of claims 35 to 37, wherein the volume of the pleural effusion is about 90% less than the volume of the pleural effusion prior to administration of thiostrepton.

41. A pharmaceutical composition comprising thiostrepton and at least one pharmaceutically acceptable carrier, wherein the pharmaceutical composition comprises about 50 mg to about 500 mg of thiostrepton.

42. The pharmaceutical composition of claim 41, wherein the composition comprises about 90 mg to about 450 mg of thiostrepton.

43. The pharmaceutical composition of claim 42, wherein the composition comprises about 90 mg, about 180 mg, about 270 mg, about 360 mg, or about 450 mg of thiostrepton.

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