Oral PCLX-001 in the treatment of human cancer

Abstract

There is described herein a method for treating cancer in a subject in need thereof, comprising, administering an oral dosage form of PCLX-001, wherein said oral dosage comprises about 20mg PCLX-001 per day to about 210 mg PCLX-001 per day.

Description

ORAL PCLX-001 IN THE TREATMENT OF HUMAN CANCER FIELD

[0001] The present disclosure relates generally to oral PCLX-001 in the treatment of human cancer. BACKGROUND

[0002] Myristoylation, the N-terminal modification of proteins with the fatty acid myristate, is critical for membrane targeting of several hundred human protein, including ones critical for intracellular signaling. Because cancer cells often have increased N- myristoyltransferase (NMT) protein expression, NMTs have been proposed as anti-cancer targets. SUMMARY

[0003] In one aspect there is provided a method for treating cancer in a human subject in need thereof, comprising, administering an oral dosage form of PCLX-001, wherein said oral dosage comprises about 20mg PCLX-001 per day to about 210 mg PCLX- 001 per day, wherein said oral dosage provides an effect comprising a Tmax of about 1 hours to about 4 hours or wherein said oral dosage provides an effect comprising a Tmax of about 1 hours to about 6 hours if said subject is taking a proton pump inhibitor (PPI).

[0004] In one example, said oral dosage comprises about 20mg PCLX-001 per day, 40mg PCLX-001 per day, 70mg PCLX-001 per day, 100mg PCLX-001 per day, 140mg PCLX-001 per day, or 210mg PCLX-001 per day.

[0005] In one example, said administration is daily administration from 1 to 28 days.

[0006] In one example, the cancer is ovarian cancer, refractory ovarian cancer, colorectal adenocarcinoma, or metastatic colorectal cancer.

[0007] In one example, the cancer is Diffuse Large B Cell Lymphoma (DLBCL).

[0008] In one example, said DLBC is refractory DLBC.

[0009] In one example, the cancer is lymphoma, leukemia, myeloma, breast cancer, small cell lung cancer, non-small cell lung cancer cells; melanoma, adenocarcinoma, pancreas cancer, bladder cancer, ovarian cancer, brain cancer, colon cancer, rectal cancer,colorectal cancer, neuroblastoma, carcinoma, endometrial cancer, sarcoma, thyroid cancer, fibrosarcoma, oral cancer, tongue cancer, nasopharynx cancer, larynx cancer, esophageal cancer, germ cell cancer, stomach cancer, hepatocellular cancer, biliary tract cancer, small intestinal cancer, testis cancer, epidermoid cancer, cervical cancer, kidney cancer, prostate cancer, or pharynx cancer.

[0010] In one aspect there is provided a use of an oral dosage of PCLX-001 for treating cancer in a human subject, wherein said oral dosage comprises about 20mg PCLX- 001 per day to about 210 mg PCLX-001 per day, wherein said oral dosage provides an effect comprising a Tmax of about 1 hours to about 4 hours or wherein said oral dosage provides an effect comprising a Tmax of about 1 hours to about 6 hours if said subject is taking a proton pump inhibitor (PPI).

[0011] In one aspect there is provided a use of an oral dosage of PCLX-001 in the manufacture of a medicament for treating cancer in a subject, wherein said oral dosage comprises about 20mg PCLX-001 per day to about 210 mg PCLX-001 per day, wherein said oral dosage provides an effect comprising a Tmax of about 1 hours to about 4 hours or wherein said oral dosage provides an effect comprising a Tmax of about 1 hours to about 6 hours if said subject is taking a proton pump inhibitor (PPI).

[0012] In one example, said oral dosage comprises about 20mg PCLX-001 per day, 40mg PCLX-001 per day, 70mg PCLX-001 per day, 100mg PCLX-001 per day, 140mg PCLX-001 per day, or 210mg PCLX-001 per day.

[0013] In one example, the cancer is ovarian cancer, refractory ovarian cancer, colorectal adenocarcinoma, or metastatic colorectal cancer.

[0014] In one example, the cancer is Diffuse Large B Cell Lymphoma (DLBCL).

[0015] In one example, said DLBC is refractory DLBC.

[0016] In one example, the cancer is lymphoma, leukemia, myeloma, breast cancer, small cell lung cancer, non-small cell lung cancer cells; melanoma, adenocarcinoma, pancreas cancer, bladder cancer, ovarian cancer, brain cancer, colon cancer, rectal cancer, colorectal cancer, neuroblastoma, carcinoma, endometrial cancer, sarcoma, thyroid cancer, fibrosarcoma, oral cancer, tongue cancer, nasopharynx cancer, larynx cancer, esophageal cancer, germ cell cancer, stomach cancer, hepatocellular cancer, biliary tract cancer, smallintestinal cancer, testis cancer, epidermoid cancer, cervical cancer, kidney cancer, prostate cancer, or pharynx cancer. . BRIEF DESCRIPTION OF THE FIGURES

[0017] Embodiments of the present disclosure will now be described, by way of example only, with reference to the attached Figures.

[0018] Figure 1 depicts the chemical structure of PCLX-001 (2,6-dichloro-N-(3- isobutyl-1,5-dimethyl-1H-pyrazol-4-yl)-4-(2- (piperazin-1-yl)pyridin-4-yl)benzene sulfonamide) C24H30Cl2N6O2S. DETAILED DESCRIPTION

[0019] In one aspect there is provided a method of treating cancer in a subject, comprising: administering an oral dosage form of PCLX-001.

[0020] PCLX-001 is also known as DDD86418 (WO 2010 / 026365) and Zelenirstat.

[0021] In one aspect there is provided a method of treating cancer in a subject, comprising: administering an oral dosage form of PCLX-001, wherein said oral dosage comprises from about 20mg PCLX-001 per day to about 210mg PCLX-001 per day. In a specific example, the oral dosage is about 210mg PCLX-001 per day. In another specific examples, the oral dosage is less than 280mg PCLX-001 per day. In another specific example, the oral dosage is less than about or equal to about 210mg PCLX-001 per day.

[0022] In some examples, the oral dosage of PCLX-001 is administered about 28 days. In some examples, the oral dosage of PCLX-001 is administered more than about 28 days. In some examples, the oral dosage of PCLX-001 is administered less than about 28 days.

[0023] The term “cancer”, as used herein, refers to a variety of conditions caused by the abnormal, uncontrolled growth of cells. Cells capable of causing cancer, referred to as “cancer cells”, possess characteristic properties such as uncontrolled proliferation, immortality, metastatic potential, rapid growth and proliferation rate, and / or certain typical morphological features. Cancer cells may be in the form of a tumor, but such cells may also exist alone within a subject, or may be a non-tumorigenic cancer cell.

[0024] A cancer may be detected in any of a number of ways, including, but not limited to, detecting the presence of a tumor or tumors (e.g., by clinical or radiological means),examining cells within a tumor or from another biological sample (e.g., from a tissue biopsy), measuring blood markers indicative of cancer, and detecting a genotype indicative of a cancer. However, a negative result in one or more of the above detection methods does not necessarily indicate the absence of cancer, e.g., a patient who has exhibited a complete response to a cancer treatment may still have a cancer, as evidenced by a subsequent relapse.

[0025] It will be appreciated that, in general, determination of the severity of disease requires identification of certain disease characteristics, for example, whether the cancer is pre-metastatic or metastatic, the stage and / or grade of cancer, and the like.

[0026] Staging is a process used to describe how advanced a cancer is in a subject. Staging may be important in determining a prognosis, planning treatment and evaluating the results of such treatment. While different cancer staging systems may need to be used for different types of cancer, most staging systems generally involve describing how far the cancer has spread anatomically and attempt to put subjects with similar prognosis and treatment in the same staging group.

[0027] Examples of common staging systems used for most solid tumours, some leukemias and lymphomas are the Overall Stage Grouping system and the ΤΜΝ system. In the Overall Stage Grouping system, Roman numerals I through IV are utilized to denote the four stages of a cancer. Generally, if a cancer is only detectable in the area of the primary lesion without having spread to any lymph nodes it is called Stage I. Stage II and III cancers are generally locally advanced and / or have spread to the local lymph nodes. For example, if the cancer is locally advanced and has spread only to the closest lymph nodes, it is called Stage II. In Stage III, the cancer is locally advanced and has generally spread to the lymph nodes in near proximity to the site of the primary lesion. Cancers that have metastasized from the primary tumour to a distant part of the body, such as the liver, bone, brain or another site, are called, Stage IV, the most advanced stage. Accordingly, stage I cancers are generally small localized cancers that are curable, while stage IV cancers usually represent inoperable or metastatic cancers. As with other staging systems, the prognosis for a given stage and treatment often depends on the type of cancer. For some cancers, classification into four prognostic groups is insufficient and the overall staging is further divided into subgroups. In contrast, some cancers may have fewer than four stage groupings.

[0028] A cancer that recurs after all visible tumour has been eradicated is called recurrent disease, with local recurrence occurring in the location of the primary tumour and distant recurrence representing distant metastasis.

[0029] Variations to the staging systems may depend on the type of cancer. Moreover, certain types of cancers. The staging system for individual cancers maybe revised with new information and subsequently, the resulting stage may change the prognosis and treatment for a specific cancer.

[0030] The “grade” of a cancer may be used to describe how closely a tumour resembles normal tissue of its same type. Based on the microscopic appearance of a tumour, pathologists identify the grade of a tumour based on parameters such as cell morphology, cellular organization, and other markers of differentiation. As a general rule, the grade of a tumour corresponds to its rate of growth or aggressiveness and tumours are typically classified from the least aggressive (Grade I) to the most aggressive (Grade IV).

[0031] Accordingly, the higher the grade, the more aggressive and faster growing the cancer. Information about tumour grade is useful in planning treatment and predicting prognosis.

[0032] In some examples, in the case of lymphoma, Stage 1 refers to lymphoma in only one group of lymph nodes. Stage II refers to two or more groups of lymph nodes are affected but they are all either above or below the diaphragm, either all in the chest or all in the abdomen. Stage III refers to two or more groups of lymph nodes are affected in both the chest and the abdomen. Stage IV refers to lymphoma is in at least one organ (e.g., bone marrow, liver or lungs) as well as the lymph nodes. Additional designations may be added to the foregoing stages. For example, “A” generally means the patient has not experiences any troublesome symptoms. “B” means the patient has experienced B symptoms (e.g., fever, night sweats, weight loss). X means the patient has bulky disease (e.g., large tumour greater than 10cm in size). E means the patient has extranodal disease (e.g., disease outside the lymph nodes).

[0033] In a specific example, the cancer is a lymphoma.

[0034] The term “lymphoma” generally refers to a malignant neoplasm of the lymphatic system, including cancer of the lymphatic system. The two main types of lymphoma are Hodgkin’s disease (HD or HL) and non-Hodgkin’s lymphoma (NHL). Abnormal cells appear ascongregations which enlarge the lymph nodes, form solid tumours in the body, or more rarely, like leukemia, circulate in the blood. Hodgkin’s disease lymphomas, include nodular lymphocyte predominance Hodgkin’s lymphoma; classical Hodgkin’s lymphoma; nodular sclerosis Hodgkin’s lymphoma; lymphocyterich classical Hodgkin’s lymphoma; mixed cellularity Hodgkin’s lymphoma; lymphocyte depletion Hodgkin’s lymphoma. Non-Hodgkin’s lymphomas include small lymphocytic NHL, follicular NHL; mantle cell NHL; mucosa- associated lymphoid tissue (MALT) NHL; diffuse large cell B-cell NHL; mediastinal large B-cell NHL; precursor Τ lymphoblastic NHL; cutaneous T-cell NHL; T-cell and natural killer cell NHL; mature (peripheral) T-cell NHL; Burkitt’s lymphoma; mycosis fungoides; Sezary Syndrome; precursor B-lymophoblastic lymphoma; B-cell small lymphocytic lymphoma; lymphoplasmacytic lymphoma; splenic marginal zone B-cell lymphoma; nodal marginal zone lymphoma; plasma cell myeloma / plasmacytoma; intravascular large B-cell NHL; primary effusion lymphoma; blastic natural killer cell lymphoma; enteropathy-type T-cell lymphoma; hepatosplenic gamma-delta T-cell lymphoma; subcutaneous panniculitis-like T-cell lymphoma; angioimmunoblastic Τcell lymphoma; primary systemic anaplastic large T / null cell lymphoma, diffuse large B-cell lymphoma (DLBCL), refractory diffuse large B-cell lymphoma (DLBCL).

[0035] In a specific example, the caner is ovarian cancer. In another specific example, the caner is refractory ovarian cancer.

[0036] In a specific example, the cancer is colorectal adenocarcinoma. In another specific example the cancer is metastatic colorectal adenocarcinoma.

[0037] In a specific example, the lymphoma is a B-cell lymphoma.

[0038] In one example, the cancer is diffuse large B-cell lymphoma (DLBCL).

[0039] In one example, the cancer is refractory diffuse large B-cell lymphoma (DLBCL).

[0040] In some examples, the compositions and / or compositions described herein (for example, PCLX-001) may be used to treat various stages and grades of cancer development and progression. In some examples, PCLX-001 may be used in the treatment of early stage cancers including early neoplasia that may be small, slow growing, localized and / or nonaggressive, for example, with the intent of curing the disease or causing regression of the cancer, as well as in the treatment of intermediate stage and in the treatment of late stage cancers including advanced and / or metastatic and / or aggressive neoplasia, for example, to slow the progression of the disease, to reduce metastasis or to increase the survival of thepatient. Similarly, PCLX-001 may be used in the treatment of low grade cancers, intermediate grade cancers and or high grade cancers.

[0041] In some examples, it is contemplated that PCLX-001 may be used in the treatment of indolent cancers, recurrent cancers including locally recurrent, distantly recurrent and / or refractory cancers (i.e., cancers that have not responded to treatment), metastatic cancers, locally advanced cancers and aggressive cancers.

[0042] In another example, the cancer is leukemia, myeloma, breast cancer, small cell lung cancer, non-small cell lung cancer cells; melanoma, adenocarcinoma, pancreas cancer, bladder cancer, ovarian cancer, brain cancer, colon cancer, neuroblastoma, carcinoma, endometrial cancer, sarcoma, thyroid cancer, fibrosarcoma, oral cancer, tongue cancer, nasopharynx cancer, larynx cancer, esophageal cancer, germ cell cancer, stomach cancer, hepatocellular cancer, biliary tract cancer, small intestinal cancer, testis cancer, epidermoid cancer, cervical cancer, kidney cancer, prostate cancer, or pharynx cancer

[0043] In some examples, PCLX-001 may be used alone or in combination with one or more therapeutic agents as part of a primary therapy or an adjuvant therapy. “Primary therapy” or “first-line therapy” refers to treatment upon the initial diagnosis of cancer in a subject. Exemplary primary therapies may involve surgery, a wide range of chemotherapies, immunotherapy and / or radiotherapy. When first-line or primary therapy is not systemic chemotherapy or immunotherapy, then subsequent chemotherapy or immunotherapy may be considered as “first-line systemic therapy”. In one example, PCLX-001 may be used for first- line systemic therapy.

[0044] The term “adjuvant therapy” refers to a therapy that follows a primary therapy and that is administered to subjects at risk of relapsing. Adjuvant systemic therapy is typically begun soon after primary therapy to delay recurrence, prolong survival or cure a subject. Treatment of a refractory cancer may be termed a “second-line therapy” and is a contemplated use of the present invention, in addition to first-line therapy.

[0045] In some examples, a subject is also treated with a proton pump inhibitor (PPI). PPIs are typically prescribed for short-term treatment of active duodenal ulcers, gastrointestinal ulcers, gastroesophageal reflux disease (GERD), severe erosive esophagitis, and / or poorly responsive symptomatic GERD.

[0046] Examples of proton pump inhibitors include, omeprazole (Prilosec®), lansoprazole (Prevacid®), esomeprazole (Nexium®), rabeprazole (Aciphex®), pantoprazole (Protonix®), pariprazole, tenatoprazole, and leminoprazole. The drugs of this class suppress gastrointestinal acid secretion by the specific inhibition of the H+ / K+-ATPase enzyme system (proton pump) at the secretory surface of the gastrointestinal parietal cell.

[0047] In a specific examples, the PPI used is pantoprazole.

[0048] In one embodiment of the present invention, the combinations are used in the treatment of an early stage cancer. In another embodiment, the combinations are used as a first-line systemic therapy for an early stage cancer.

[0049] In an alternate example, PCLX-001 may be used in the treatment of a late stage and / or advanced and / or metastatic cancer. In a further embodiment, PCLX-001 may be administered as a first-line systemic therapy for the treatment of a late stage and / or advanced and / or metastatic cancer.

[0050] A refractory cancer refers to a cancer that is not amenable to surgical intervention and the cancer is either initially unresponsive to chemo- or radiation therapy or the cancer becomes unresponsive over time.

[0051] The term “relapse” refers to those patients who achieved a clinical response (CR) or partial response (PR) and subsequently experienced disease progression.

[0052] The term “clinical response” refers to inhibition of disease progression, inhibition of tumor growth, reduction of primary tumor, relief of tumor-related symptoms, inhibition of tumor secreted factors (including tumor secreted hormones, such as those that contribute to carcinoid syndrome), delayed appearance of primary or secondary tumors, slowed development of primary or secondary tumors, decreased occurrence of primary or secondary tumors, slowed or decreased severity of secondary effects of disease, arrested tumor growth and regression of tumors, increased Time To Progression (TTP), increased Progression Free Survival (PFS), increased Overall Survival (OS), among others.

[0053] The term “overall survival” or “OS” refers to the time from enrollment to death from any cause.

[0054] The term “duration of response” or “DOR” refers to the time from the first occurrence of a documented objective response to disease progression.

[0055] The term “progression free survival” or “PFS” refers to the time from enrollment to the date of the first recorded occurrence of disease progression.

[0056] The term “stable disease” or “SD” refers to neither sufficient shrinkage to qualify for CR or PR nor sufficient increase growth of tumor to qualify for PD.

[0057] The term “partial response” refers to persistence of one or more non-target lesions and / or (if applicable) maintenance of tumor marker level above the normal limits.

[0058] The term “subject”, as used herein, refers to an animal, and can include, for example, domesticated animals, such as cats, dogs, etc., livestock (e.g., cattle, horses, pigs, sheep, goats, etc.), laboratory animals (e.g., mouse, rabbit, rat, guinea pig, etc.), mammals, non-human mammals, primates, non-human primates, rodents, birds, reptiles, amphibians, fish, and any other animal.

[0059] In a specific example, the subject is a human.

[0060] The term “treatment” or “treat” as used herein, refers to obtaining beneficial or desired results, including clinical results. Beneficial or desired clinical results can include, but are not limited to, alleviation or amelioration of one or more symptoms or conditions, diminishment of extent of disease, stabilized (i.e. not worsening) state of disease, preventing spread of disease, delay or slowing of disease progression, amelioration or palliation of the disease state, diminishment of the reoccurrence of disease, and remission (whether partial or total), whether detectable or undetectable. "Treating" and "Treatment" can also mean prolonging survival as compared to expected survival if not receiving treatment. "Treating" and "treatment" as used herein also include prophylactic treatment. For example, a subject with early cancer, for example an early stage lymphoma, can be treated to prevent progression or alternatively a subject in remission can be treated with a compound or composition described herein to prevent recurrence.

[0061] The term "pharmaceutically acceptable" as used herein includes compounds, materials, compositions, and / or dosage forms (such as unit dosages) which are suitable for use in contact with the tissues of a subject without excessive toxicity, irritation, allergic response, or other problem or complication, commensurate with a reasonable benefit / risk ratio. Each carrier, excipient, etc. is also be "acceptable" in the sense of being compatible with the other ingredients of the formulation.

[0062] The term "excipient" means a pharmacologically inactive component such as a diluent, lubricant, surfactant, carrier, or the like. Excipients that are useful in preparing a pharmaceutical composition are generally safe, non-toxic and are acceptable for human pharmaceutical use. Reference to an excipient includes both one and more than one such excipient.

[0063] As used herein, the term "pharmaceutically acceptable carrier" refers to any of the standard pharmaceutical carriers including, but not limited to, phosphate buffered saline solution, water, emulsions (e.g., such as an oil / water or water / oil emulsions), and various types of wetting agents, any and all solvents, dispersion media, coatings, sodium lauryl sulfate, isotonic and absorption delaying agents, disintegrants (e.g., potato starch or sodium starch glycolate), stabilizers and preservatives, and the like.

[0064] A "treatment or dosage regimen" as used herein refers to a combination of dosage, frequency of administration, or duration of treatment, with or without addition of a second medication.

[0065] The term “diagnosis” as used herein, refers to the identification of a molecular and / or pathological state, disease or condition, such as the identification of lymphoma, or other type of cancer.

[0066] The term "alleviates" as used herein refers to a decrease, reduction or elimination of a condition, disease, disorder, or phenotype, including an abnormality or symptom.

[0067] In some aspects there is provided a pharmaceutical composition comprising PCLX-001 in admixture with a pharmaceutically acceptable adjuvant, diluent or carrier.

[0068] Pharmaceutical compositions of the present disclosure may also contain adjuvants such as preservative, wetting agents, emulsifying agents and dispersing agents. Prevention of the action of microorganisms may be ensured by the inclusion of various antibacterial and antifungal agents, for example, paraben, chlorobutanol or phenol sorbic acid. It may also be desirable to include isotonic agents such as sugars or sodium chloride, for example.

[0069] Solid dosage forms for oral administration include capsules, tablets, pills, powders and granules. In such solid dosage forms, an active compound (e.g., PCLX-001) is typically mixed with at least one inert, pharmaceutically acceptable excipient or carrier such assodium citrate or dicalcium phosphate and / or one or more: a) fillers or extenders such as starches, lactose, sucrose, glucose, mannitol and silicic acid; b) binders such as carboxymethylcellulose, alginates, gelatin, polyvinylpyrrolidone, sucrose and acacia; c) humectants such as glycerol; d) disintegrating agents such as agar-agar, calcium carbonate, potato or tapioca starch, alginic acid, certain silicates and sodium carbonate; e) solution retarding agents such as paraffin; f) absorption accelerators such as quaternary ammonium compounds; g) wetting agents such as cetyl alcohol and glycerol monostearate; h) absorbents such as kaolin and bentonite clay and i) lubricants such as talc, calcium stearate, magnesium stearate, solid polyethylene glycols, sodium lauryl sulfate and mixtures thereof. In the case of capsules, tablets and pills, the dosage form may also comprise buffering agents. Solid compositions of a similar type may also be employed as fillers in soft and hard-filled gelatin capsules using such excipients as lactose or milk sugar as well as high molecular weight polyethylene glycol, for example.

[0070] The term “solid” as used herein refers to a substance that is solid or semi-solid at room temperature. Hence, as used herein, a “solid” substance may become liquid at, e.g., body temperature.

[0071] Cmax is the maximum plasma concentration of PCLX-001.

[0072] Tmax is the time at which the maximum plasma concentration of PCLX-001 is achieved.

[0073] The term “pharmacokinetic” (or PK) refers to the factors which determine the attainment and maintenance of the appropriate concentration of drug at a site of action.

[0074] The term "AUC" refers to the area under concentration (or area under the curve) representing the concentration of a compound or metabolite thereof in a biological fluid of a patient as a function of time following administration of the compound to the patient.

[0075] In vitro studies of PCLX-001 have shown inhibition of cell growth of hematological cancer cells, including lymphomas, leukemia, and myeloma; breast cancer cells; small cell lung cancer cells; non-small cell lung cancer cells; melanoma cells; adenocarcinoma cells; pancreas cancer cells; bladder cancer cells; ovary cancer cells; brain cancer cells; colon cancer cells; neuroblastoma; carcinoma cancer cells; endometrial cancer cells; sarcoma cancer cells; thyroid cancer cells; Fibrosarcoma cancer cells; tongue cancer cells; testis cancercells; epidermoid cancer cells; cervix cancer cells; kidney cancer cells; prostate cancer cells; and pharynx cancer cells.(3)

[0076] The term “bioavailability" refers to the rate and amount of a drug that reaches the systemic circulation of a patient following administration of the drug or prodrug thereof to the patient and can be determined by evaluating, for example, the plasma or blood concentration-versus-time profile for a drug. Parameters useful in characterizing a plasma or blood concentration-versus-time curve include the area under concentration (AUC), the time to peak concentration (Tmax), and the maximum drug concentration (Cmax).

[0077] Absolute oral bioavailability is the bioavailability of a compound or metabolite thereof following oral administration compared to the bioavailability following intravenous administration of an equivalent amount of the compound or metabolite thereof. Relative oral bioavailability of a compound or metabolite thereof is the bioavailability following oral administration of a compound or metabolite thereof relative to administration of an equivalent amount of the compound or metabolite thereof in another dosage form and / or route of administration.

[0078] The term "Dosage form" refers to a form of a formulation that contains an amount of active agent or prodrug of an active agent, e.g., PCLX-001, which may be administered to a patient to achieve a therapeutic effect.

[0079] An oral dosage form is intended to be administered to a patient taken orally, for example via the mouth and swallowed.

[0080] The terms “taken orally” and “oral administration” refer to a manner of providing an active agent to a subject or patient by the mouth through the gastrointestinal tract (digestive tract, digestional tract, GI tract, GIT, gut, or alimentary canal) and are used interchangeably. The gastrointestinal tract is an organ system within humans and other animals which takes in food, digests it to extract and absorb energy and nutrients, and expels the remaining waste as feces. The mouth, esophagus, stomach and intestines are part of the gastrointestinal tract.

[0081] Suitably, oral formulations may contain a dissolution aid. The dissolution aid is not limited as to its identity so long as it is pharmaceutically acceptable. Examples include non- ionic surface active agents, such as sucrose fatty acid esters, glycerol fatty acid esters, sorbitan fatty acid esters (e.g. sorbitan trioleate), polyethylene glycol, polyoxyethylene hydrogenated castor oil, polyoxyethylene sorbitan fatty acid esters, polyoxyethylene alkylethers, methoxypolyoxyethylene alkyl ethers, polyoxyethylene alkylphenyl ethers, polyethylene glycol fatty acid esters, polyoxyethylene alkylamines, polyoxyethylene alkyl thioethers, polyoxyethylene polyoxypropylene copolymers, polyoxyethylene glycerol fatty acid esters, pentaerythritol fatty acid esters, propylene glycol monofatty acid esters, polyoxyethylene propylene glycol monofatty acid esters, polyoxyethylene sorbitol fatty acid esters, fatty acid alkylolamides, and alkylamine oxides; bile acid and salts thereof (e.g. chenodeoxycholic acid, cholic acid, deoxycholic acid, dehydrocholic acid and salts thereof, and glycine or taurine conjugate thereof); ionic surface active agents, such as sodium laurylsulfate, fatty acid soaps, alkylsulfonates, alkylphosphates, ether phosphates, fatty acid salts of basic amino acids; triethanolamine soap, and alkyl quaternary ammonium salts; and amphoteric surface active agents, such as betaines and aminocarboxylic acid salts.

[0082] In one example, the tosylate salt of PCLX-001 is used.

[0083] The solid dosage forms of tablets, dragees, capsules, pills, and granules can be prepared with coatings and shells such as enteric coatings and other coatings well known in the pharmaceutical formulating art. They may optionally contain opacifying agents and may also be of a composition such that they release the active ingredient(s) only, or preferentially, in a certain part of the intestinal tract, and / or in delayed fashion. Examples of embedding compositions include polymeric substances and waxes.

[0084] PCLX-001 may also be in a micro-encapsulated form, if appropriate, with one or more of the above-mentioned excipients.

[0085] Liquid dosage forms for oral administration include pharmaceutically acceptable emulsions, solutions, suspensions, syrups and elixirs. In addition to PCLX-001, the liquid dosage forms may contain inert diluents commonly used in the art such as water or other solvents, solubilizing agents and emulsifiers such as ethyl alcohol, isopropyl alcohol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butylene glycol, dimethyl formamide, oils (in particular, cottonseed, groundnut, corn, germ, olive, castor, and sesame oils), glycerol, tetrahydrofurfuryl alcohol, polyethylene glycols and fatty acid esters of sorbitan and mixtures thereof. Besides inert diluents, the oral compositions may also include adjuvants such as wetting agents, emulsifying and suspending agents, sweetening, flavoring and perfuming agents. Suspensions, in addition to the active compounds, may contain suspending agents such as ethoxylated isostearyl alcohols, polyoxyethylene sorbitol andsorbitan esters, microcrystalline cellulose, aluminum metahydroxide, bentonite, agar-agar, and tragacanth and mixtures thereof.

[0086] In other examples, pharmaceutical compositions of the present disclosure may be used for parenteral injection suitably comprise pharmaceutically acceptable sterile aqueous or nonaqueous solutions, dispersions, suspensions or emulsions as well as sterile powders for reconstitution into sterile injectable solutions or dispersions just prior to use. Examples of suitable aqueous and nonaqueous carriers, diluents, solvents or vehicles include water, ethanol, polyols (such as glycerol, propylene glycol, polyethylene glycol and the like), 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 the maintenance of the required particle size in the case of dispersions and by the use of surfactants.

[0087] Prolonged absorption of an injectable pharmaceutical form may be brought about by the inclusion of agents (for example aluminum monostearate and gelatin) which delay absorption.

[0088] In some cases, in order to prolong the effect of the pharmaceutical compositions of the present disclosure, it is desirable to slow the absorption of the composition, or PCLX- 001 from subcutaneous or intramuscular injection. This may be accomplished by the use of a liquid suspension, where PCLX-001 is suspended in. The rate of absorption of PCLX-001 then depends upon its rate of dissolution.

[0089] Alternatively, delayed absorption of a parenterally administered pharmaceutical composition of the present disclosure is accomplished by dissolving or suspending PCLX-001 in an oil vehicle. Injectable depot forms are suitably made by forming microencapsulated matrices of PCLX-001 in biodegradable polymers, for example polylactide-polyglycolide. Depending upon the ratio of PCLX-001 to polymer and the nature of the particular polymer employed, the rate of release can be controlled. Examples of other biodegradable polymers include poly(orthoesters) and poly(anhydrides).

[0090] Depot injectable formulations may also be prepared by entrapping PCLX-001 in liposomes or microemulsions which are compatible with body tissues. The injectable formulations can be sterilized, for example, by filtration through a bacterial- retaining filter orby incorporating sterilizing agents in the form of sterile solid compositions which can be dissolved or dispersed in sterile water or other sterile injectable media just prior to use.

[0091] In other examples, pharmaceutical compositions of the present disclosure for rectal or vaginal administration are preferably suppositories which can be prepared by mixing PCLX-001 with suitable non-irritating excipients or carriers such as cocoa butter, polyethylene glycol or a suppository wax which are solid at room temperature but liquid at body temperature and therefore melt in the rectum or vaginal cavity and release PCLX-001.

[0092] PCLX-001 may also be administered in the form of liposomes. As is known in the art, liposomes are generally derived from phospholipids or other lipid substances. Liposomes are formed by mono- or multi-lamellar hydrated liquid crystals which are dispersed in an aqueous medium. Any non-toxic, physiologically acceptable and metabolizable lipid capable of forming liposomes can be used. Pharmaceutical compositions of the present disclosure in liposome form can contain, in addition to PCLX-001, stabilisers, preservatives, excipients and the like. The preferred lipids are the phospholipids and the phosphatidyl cholines (lecithins), both natural and synthetic. Methods to form liposomes are known in the art.

[0093] Dosage forms for topical administration of PCLX-001include powders, sprays, ointments and inhalants. PCLX-001 may be mixed under sterile conditions with a pharmaceutically acceptable carrier and any needed preservatives, buffers or propellants which may be required. PCLX-001 may also be administered, for example as part of a pharmaceutical composition, in ophthalmic formulations, or eye ointments, powders and solutions.

[0094] Method of the invention are conveniently practiced by providing the compounds and / or compositions used in such method in the form of a kit. Such kit preferably contains the composition. Such a kit preferably contains instructions for the use thereof.

[0095] To gain a better understanding of the invention described herein, the following examples are set forth. It should be understood that these examples are for illustrative purposes only. Therefore, they should not limit the scope of this invention in anyway.

[0096] EXAMPLES

[0097] Introduction

[0098] The two human N-myristoyltransferases, NMT1 and NMT2, add a fourteen- carbon fatty acid, myristate, to the N terminus of other proteins, stabilizing them from degradation and targeting them to intracellular membranes (1). More than 300 human proteins undergo myristoylation (1). These include key proteins that regulate cell growth and apoptosis, including Src, and seven of the human Src Family tyrosine Kinases (SFKs; Blk, Fgr, Fyn, Hck, Lck, Lyn, and Yes). Aberrant myristoylation has been identified in many common human cancers (2,3,4), and inhibitors of NMT have been proposed as potential anti- cancer therapeutics (2).

[0099] In vitro preclinical studies with zelenirstat (also known as PCLX-001 and DDD86481), a potent, small molecule inhibitor of both human NMT proteins, induced dose- and time-dependent apoptosis in cancer cells at concentrations lower than that required to kill normal cells. Zelenirstat was cytotoxic or cytostatic across a wide variety of cultured human cancer cells, with sensitivity noted in a broad array of hematologic malignancies including B-cell NHL, AML, and multiple myeloma (3). Comparative transcriptomic analysis of zelenirstat sensitive and insensitive cell lines identified an expression signature, MISS-91, that predicts those cancers most likely to be responsive to NMT inhibitor therapy (7,8). The cancer types with the highest sensitivity scores included Diffuse Large B-cell lymphoma (DLBCL), acute myelogenous leukemia (AML), colorectal adenocarcinoma, testicular germ cell tumors, cervical squamous cell carcinomas, squamous cell carcinoma of the lung, basal subtype breast adenocarcinoma, and ovarian carcinoma.

[0100] In mice-bearing established human tumor xenografts, zelenirstat achieved tumor growth inhibition, but no regression, in cell line derived xenografts from multiple solid tumors (Pacylex, data on file). However, profound tumor regressions and complete remissions were seen in patient derived xenograft models of refractory Diffuse Large B-Cell lymphoma, where loss of Lyn and HGAL, which require myristylation to function, inhibited B- cell receptor signaling (3). Zelenirstat produced similarly profound remissions in patient derived xenograft models of acute myelogenous leukemia. In murine tail-vein injection models of human AML, zelenirstat reduced the leukemic cell burden and dramatically reduced the human leukemic stem cell population (8a).

[0101] We conducted formal toxicologic evaluation in rats and dogs and found acute gastrointestinal and hematologic toxicities at the highest doses examined. With daily oral dosing at or slightly above the maximally tolerated dose, diarrhea, dehydration and weight loss were rapidly reversible with discontinuation of zelenirstat and did not recur when re- introduced at lower doses (5). No cumulative toxicities were identified.

[0102] The highly novel mechanisms of action, the promising pre-clinical findings, and the potential for a convenient oral therapy prompted the advancement of zelenirstat into clinical testing. The principal objectives of this phase I, first in human, and pharmacokinetic study of zelenirstat were to determine (i) the maximum-tolerated dose (MTD) / recommended phase II dose, (ii) safety, and (iii) the pharmacokinetic characteristics of daily oral zelenirstat in patients with refractory solid tumors and relapsed B-cell NHL.

[0103] Purpose

[0104] Myristoylation regulates numerous membrane-bound signal transduction pathways important in cancer biology, including Src and Src family tyrosine kinases. This modification is catalyzed by N-myristoyltransferases 1 and 2 (NMT1 and NMT2). Zelenirstat is an orally available small molecule potent NMT inhibitor of both NMT proteins. The primary objective of this phase I multiple ascending dose study is to determine the safety, tolerability, and pharmacokinetics MTD and / or recommended phase II dose, of zelenirstat as monotherapy in patients with refractory lymphomas and advanced solid tumors.

[0105] Patients and Methods

[0106] Twenty six patients with advanced solid malignancies or lymphoma were treated with escalating doses of daily zelenitrstat administered as oral capsules in 28 day cycles. Escalation proceeded using a 3 + 3 design. Plasma samples were assayed to determine pharmacokinetic parameters.

[0107] Results

[0108] Between September 2021 and July 2023, twenty-six patients received sixty- three 28-day cycles of zelenirstat at doses ranging from 20 mg / d to 280 mg / d. Medication compliance was high. Dose limiting toxicities were not observed up to and including the 210 mg / d cohort, but dose limiting gastrointestinal toxicities were seen in the 280 mg / d cohort, establishing 210 mg / d to be the Maximally Tolerated Dose (MTD). In doses up to and including 210 mg / d, the most common treatment-emergent all-grade adverse events weregastrointestinal disorders, reported in 29% of patients; these were primarily low-grade diarrhea, nausea or vomiting. Transient grade II thrombocytopenia was noted in three patients. No other adverse events were seen at a frequency of greater than five percent. There was no evidence of neuropathy, alopecia, or QT interval prolongation induced by zelenirstat.

[0109] Plasma concentrations peaked between 1 and 4 hours across the dose cohorts, with terminal half-life values ranging from 6.7 to 12 hours. Drug exposure tended to be lower in patients on concomitant proton pump inhibitors, which were prohibited in the higher dose cohorts. Steady state was achieved by day 8 to 15 in most patients, and trough concentrations exceeded the levels predicted to be therapeutic in the higher dose cohorts.

[0110] Seventeen patients were evaluable for formal response assessment. Stable disease as best response was seen in five (30%) patients, including three solid tumor patients treated at 210 mg / day, two of whom had reductions in serum tumor markers.

[0111] Conclusion

[0112] Zelenirstat was well-tolerated on a daily oral schedule at doses as high as 210 mg daily. This study established the zelenirstat MTD to be 210 mg daily by continuous oral administration.

[0113] The absence of severe toxicities, attainment of plasma concentrations highly active in preclinical models, and early evidence of antitumor activity warrant further disease- directed studies of this agent alone and in combination with other agents in NHL, AML, and selected solid tumors.

[0114] Materials and Methods

[0115] Study Design

[0116] This phase I, multicenter, nonrandomized, open-label study of zelenirstat is entitled “Phase I Trial of PCLX-001 in B-cell Non-Hodgkin Lymphoma and Advanced Solid Malignancies” and registered at clinicaltrials.gov as NCT04836195. The study was conducted according to the guidelines of the Declaration of Helsinki, approved by Health Canada and the relevant institutional review boards in each of the four study centres. Written informed consent was obtained from all study subjects.

[0117] The study was designed in two parts, a dose escalation / multiple ascending dose phase I, reported here, and phase IIa dose expansion cohorts, now ongoing in patients with relapsed B-cell NHL, and planned for selected advanced solid tumors.

[0118] A primary endpoint of this dose escalation study is to determine the recommended dose of zelenirstat for the dose expansion phases of the trial. Using a standard 3+3 design, the recommended dose was to be the dose level below that for the cohort in which maximum tolerated dose (MTD) was reached. The MTD will have been reached when 2 or more patients in a cohort experience a first cycle dose-limiting toxicity (DLT).

[0119] Patient population

[0120] Eligible patients were 18 or older and had i) advanced solid tumor with progression after at least one prior therapy and were not eligible for therapies expected to provide clinical benefit, and ii) B-cell lymphomas including DLBCL, HGBL, FL (grades 1 to 3b), MCL, and Burkitt lymphoma who have failed at least two prior therapies and were not eligible for therapies expected to provide clinical benefit. Patients required evaluable or measurable disease, ECOG performance status of 0 or 1, and adequate bone marrow, hepatic, renal, and cardiac function. All other anticancer systemic therapies were required to be discontinued for at least three weeks, and to be off corticosteroids above prednisone equivalents of 10 mg / day. Strong CYP3A4 inhibitors and inducers were prohibited.

[0121] The investigational agent

[0122] Zelenirstat is a potent, small molecule inhibitor of human NMT1 and NMT2 proteins. In preclinical models, it has complete oral bioavailability, is highly plasma protein bound, and is metabolized in the liver primarily by CYP3A4 to water soluble metabolites excreted by the kidney (Pacylex, data on file). Zelenirstat has no relevant off-target kinase inhibition [3]. In GLP non-clinical safety testing, diarrhea was dose limiting in multiple daily oral administration, and hematologic toxicity was seen at the highest doses [5].

[0123] Drug administration

[0124] Patients received daily oral zelenirstat on 28-day cycles, escalated as per Table 2. The medication was administered as 10 mg and 70 mg capsules, with instructions to take on an empty stomach each morning with 250 ml of tapwater.

[0125] Pretreatment and Follow-Up Studies

[0126] A complete history, physical examination, and routine laboratory studies, including CBC, WBC, chemistry including electrolytes, BUN, creatinine, uric acid, glucose, alkaline phosphatase, lactate dehydrogenase, ALT, AST, total bilirubin, calcium, total protein, albumin, cholesterol, triglycerides, HIV and hepatitis B / C serologic studies, triplicate ECGs, relevant radiologic studies, and (optional) tumor markers and urinalysis were performed pre- treatment. During the study, radiologic studies for disease status were repeated at the end of every even numbered cycle. Response was assessed by Response Evaluation Criteria in Solid Tumors version 1.1 (reference) for solid tumors, and Lugano Response Criterion for NHL (reference Cheson 2014)

[0127] Plasma pharmacokinetic sampling, assay, and analyses

[0128] Peripheral blood samples were collected over six time points in the first 8 hours of cycle 1 day 1, and again on cycle 1 day 15, with pre-treatment day 1 levels obtained with every subsequent cycle. Zelenirstat plasma concentrations were quantified using a validated ultra-performance liquid chromatography with tandem mass spectrometry detection and analyzed using a nonlinear mixed-effects model.

[0129] Results

[0130] The results of the study will be discussed below under General and specific aspects of the study such as Safety, Pharmacokinetic Analysis, and others.

[0131] General

[0132] Between September 14, 2021 and 12 July, 2023, 26 patients were treated with zelenirstat at doses of 20, 40, 70, 100, 140, 210, and 280 mg / d. Patients who did not complete the first 28-day cycle of therapy with at least 21 days of treatment for reasons other than DLT were deemed not assessable and were replaced. This report was triggered by sufficient safety assessment data available to establish 210 mg / d as the maximally tolerated dose of continuous daily zelenirstat, the determination of the nature of dose-limiting toxicities, and achievement of systemic exposures projected to exceed that required for activity in selected solid tumors, B-cell lymphoma, and acute myelogenous leukemia.

[0133] Twenty-six patients, whose pertinent characteristics are listed in Table 1, received a total of sixty-three cycles of zelenirstat at doses ranging from 20 mg / d to 280 mg / d. The total number of patients treated and cycles at each dose level, as well as the overall dose-escalation scheme, are listed in Table 2.

[0134] Safety

[0135] Each subject was assessed for safety using daily diary entries, clinical assessments at baseline, days 8, 15, 22, 28, 42, 56, and every 28 days while on treatment. Dose limiting toxicities (DLTs) were defined as drug related CTCAE version 5.0 adverse events: Gr 4 platelets, Gr ≥ 3 platelets with bleeding and / or transfusions, Gr 4 ANC for ≥ 7 days, Gr ≥ 3 febrile neutropenia, and Gr ≥ 3 non-hematological toxicity.

[0136] All twenty-six treated patients were evaluable for safety endpoints. The number of cycles completed ranged from one to eight. Medication compliance was high, with patient diaries documenting 90% or higher median compliance across all dose cohorts apart from the 280 mg cohort, in which dose interruptions due to toxicity were frequent.

[0137] Across dose cohorts up to and including 210 mg, the most common treatment- emergent all-grade adverse events were gastrointestinal disorders, reported in 6 patients (29%), all of which were grade 1 or 2. Diarrhea was reported in 4 patients (19%), and nausea and vomiting in 3 patients (14%). Two patients (9%) reported GERD symptoms. All GI toxicities were self-limiting or responsive to routine supportive measures and zelenirstat continued at full dose in each case. Fatigue was reported in 4 patients (19%). Low platelets of grade 2 severity were noted in three patients (14%) but resolved despite continuation of the drug at full dose. No other drug-related adverse events were seen at a frequency of greater than five percent. No treatment emergent alopecia was observed. No neuropathy was reported. At doses up to and including 210 mg, all of the treatment emergent serious adverse events (SAEs) were attributed to progressive disease rather than drug toxicity, including one patient with pleural and pericardial mesothelioma with extensive and progressive mediastinal involvement who experienced a fatal cardiac arrest.

[0138] In the 280 mg cohort, five patients began treatment, but two discontinued therapy due to rapid clinical deterioration associated with progression of mucosal melanoma and prostatic small cell carcinoma. These patients were replaced, and two of the three subsequent patients treated at 280 mg daily experienced first cycle dose limiting gastrointestinal toxicity, including grade 3 diarrhea in the setting of biliary stent dysfunction and cholangitis, and grade 3 diverticulitis and abdominal pain in a patient with known diverticular disease.

[0139] The distributions of treatment emergent grade 3 and 4 AEs as a function of dose are listed in Table 3.

[0140] Pre and post dosing ECG assessments showed no evidence of drug-induced QT prolongation and no trend for increasing QT duration with increasing drug dose (data not shown).

[0141] Given that no DLTs were observed DLTs were observed in any of the dose escalation cohorts up to an including the 210 mg / d cohort, and that two patients experienced first cycle DLTs when treated at 280 mg / d, we established 210 mg to be the maximally tolerated dose of oral daily zelenirstat.

[0142] Pharmacokinetic Analysis

[0143] All twenty-one treated patients were included in the pharmacokinetic analysis. All patients had measurable plasma zelenirstat concentrations, with the pharmacokinetic parameters listed in Table 4.

[0144] Following single oral administration of 20 mg to 210 mg zelenirstat on Cycle 1, Day 1 (C1D1), plasma concentrations of zelenirstat steadily increased reaching peak levels with median Tmaxvalues ranging from 1 to 4 hours across the dose cohorts. After repeated daily dosing, zelenirstat Tmaxvalues on C1D15 were comparable to values obtained on C1D1. The median Tmaxrange was slightly wider (1 to 6 hours) in patients taking proton pump inhibitors (PPI) with a maximum Tmaxvalue observed of about 8 hours in those patients. After reaching peak concentrations (Cmax) on C1D1, zelenirstat was eliminated from plasma with mean terminal half-life ranging from 6.7 to 9.5 hours across the dose cohorts. Zelenirstat half-life values on C1D15 (7.67 to 12.0 hours) were comparable to values obtained on C1D1. One patient in the 40 mg cohort had prior pancreatico-duodenal resection and had very limited oral absorption of zelenirstat but was included in these analyses. Plasma concentrations (Cmaxand AUC values) tended to be lower in patients on PPIs compared to patients not taking PPIs. The lower exposure in patients on PPIs is likely due to reduced absorption of zelenirstat as half-life was roughly similar in patients on PPIs and those who were not taking PPIs.

[0145] Systemic exposure to zelenirstat (Cmaxand AUC values) tended to increase with increasing dose over the dose range of 20 mg to 210 mg. Dose-proportionality over this dose range was not supported by statistical analysis due to the limited number of subjectsand the variability introduced by the concomitant use of PPIs. The majority of patients had limited accumulation of zelenirstat (< 2-fold) following repeated administration for 14-days, with accumulation ratios for Cmax(ArCmax) and AUC0-24(ArAUC0-24) values ranging from 0.41 to 1.72 and 1.0 to 1.9, respectively. Three subjects had accumulation ratios for both (ArCmaxand ArAUC0-24) between 3 and 6-fold. Based upon pre-dose concentrations over multiple cycles, no accumulation was apparent beyond Day 15. Time to steady state was assessed by visual observation of pre-dose zelenirstat concentrations. Steady state, with some variability, was achieved by day 8 to 15 of dose administration in most patients.

[0146] Although zelenirstat is metabolized by liver to water soluble metabolites which are excreted primarily by the kidney, there were no apparent relationships noted between PK and hepatic or renal function tests. The relationships among pharmacokinetic parameters that reflect zelenirstat exposure and clinical toxicities were explored, but none were found (data not shown).

[0147] Anticancer Activity

[0148] Each subject had baseline cross sectional imaging with CT and response assessment scans on completion of every 2ndcycle. Evaluation of response was by the RECIST 1.1 criterion or the Lugano classification criteria for the evaluation of non-Hodgkin lymphoma. Nine treated patients were excluded from the efficacy cohort (one in the 20 mg cohort, one in the 70 mg cohort for progressive mediastinal mesothelioma and fatal cardiac arrest, two in the 100 mg cohort, and one in the 210 mg cohort, and 4 in the 280 mg cohort) due to lack of response assessment scans, most frequently due clinical progression or deterioration. Seventeen patients were included in the efficacy cohort, defined as those with baseline imaging and at least one follow-up imaging study. Progressive disease as the best response on trial scans was identified in twelve of seventeen patients (70%). Stable disease (SD) as best response was seen in five patients (30%). SD was noted at 15 weeks and 15 weeks in two patients treated at 40 mg and 140 mg respectively, each of whom stayed on study drug until progressive disease was identified at 24 weeks. Two patients with refractory ovarian cancer treated at 210 mg achieved SD and remained on study treatment at full dosing for six and eight cycles prior to coming off study; one of these patients had a 20% reduction in Cancer Antigen 125 (CA125). A third patient treated at 210 mg achieved SD and continues full-dose therapy beyond six cycles; this male with metastatic colonadenocarcinoma and five prior lines of chemotherapy (including flurouracil, oxaliplatin, bevacizumab, irinotecan, panitumumab, anti PD-1 monoclonal antibodies, and an investigational SIRPA antagonist) has an ongoing 13% reduction in sum of longest dimensions of the target lesions, which represents a 50% reduction in measured tumor volumes, and shows a parallel reduction in serum Carcinoembryonic Antigen (CEA) levels from baseline of approximately 50%. No objective responses were yet observed in this study, although radiologic regressions of individual target lesions were seen.

[0149] Discussion

[0150] Herein, we report a study of human exposure to any inhibitor of N- myristoyltransferase. Zelenirstat, an orally available, potent pan-NMT inhibitor was given daily in patients with refractory solid tumors and relapsed B-cell NHL. Dose dose-escalation identified gastrointestinal symptoms as the most frequent adverse events, they were not dose limiting up to doses of 210 mg daily, and responded to standard medical management without dose reduction. At doses of 280 mg daily, gastrointestinal toxicities were dose limiting. Pharmacokinetic studies confirmed rapid oral absorption, dose-dependent drug exposure, and pharmacokinetic parameters suitable for a once daily administration schedule. Zelenirstat safely achieved blood levels and exposures well above that required for cytotoxicity in cultured human cancer lines.

[0151] Efficacy was an exploratory endpoint of this study. As preclinical xenograft models suggested DLBCL was more sensitive to zelenirstat than solid tumor xenografts, and given the significant disease burden and heavily pretreated population, we did not expect to see clinical benefit in patients with solid tumours. Nonetheless, five patients with advanced solid tumors achieved stable disease as best response, and solid tumor patients treated at 210 mg daily appear to have had clinical benefit, receiving prolonged and uninterrupted durations of therapy (six and eight cycles, and a third continuing beyond six cycles) with minimal toxicity and some evidence of minor response and reductions in serum tumor markers. Ovarian cancer was among the most zelenirstat-sensitive tumor types in pre-clinical screens and in the MISS-91 analysis. Ovarian cancer metastases have higher NMT1 protein levels than paired ovarian primary cancers, and aggressive ovarian cancers have higher levels of Src, suggesting a possible mechanism for zelenirstat activity in this disease (9). Furthermore, zelenirstat has anti-angiogenic effects mediated by Src inactivation which isrequired for Vascular Endothelial Growth Factor signaling, (10), which may contribute to anticancer effects in colon cancer and other solid tumors. Zelenirstat dosing at 210 mg daily in patients with selected advanced solid tumors is ongoing to better understand the magnitude and spectrum of zelenirstat anticancer activity.

[0152] Table 1. Patient Characteristics

[0153] Table 2. Dose-Escalation Scheme

[0154] Table 3. Grade 3 or 4 Treatment Emergent Adverse Events by System Organ Class

[0155] Table 4. Summary Statistics of Plasma Pharmacokinetic Parameters

[0156] Abbreviations: AUC0-24, area under the plasma concentration-time curve;Cmax, maximum concentration; T1 / 2, half-life; Tmax, time to maximum serum concentration;PK = pharmacokinetic. N = number of participants in the PK analysis set, n = number of participants. SD = standard deviation. NC = not calculated. NA = not available

[0157] References

[0158] 1. Castrec, B., Dian, C., Ciccone, S., Ebert, C.L., Bienvenut, W.V., Le Caer, J.P., Steyaert, J.M., Giglione, C., and Meinnel, T. (2018). Structural and genomicdecoding of human and plant myristoylomes reveals a definitive recognition pattern. Nat Chem Biol 14, 671-679.

[0159] 2. Selvakumar, P., Lakshmikuttyamma, A., Shrivastav, A., Das, S.B., Dimmock, J.R., and Sharma, R.K. (2007). Potential role of N-myristoyltransferase in cancer. Prog Lipid Res 46, 1-36.

[0160] 3. Beauchamp, E., Yap, M.C., Iyer, A., Perinpanayagam, M.A., Gamma, J.M., Vincent, K.M., Lakshmanan, M., Raju, A., Tergaonkar, V., Tan, S.Y., et al. (2020). Targeting N-myristoylation for therapy of B-cell lymphomas. Nat Commun 11, 5348.

[0161] 4. Mackey, J.R., Lai, J., Chauhan, U., Beauchamp, E., Dong, W.F., Glubrecht, D., Sim, Y.W., Ghosh, S., Bigras, G., Lai, R., et al. (2021). N-myristoyltransferase proteins in breast cancer: prognostic relevance and validation as a new drug target. Breast Cancer Res Treat 186, 79-87.

[0162] 5. Weickert, M., Dillberger, J., Mackey, J.R., Wyatt, P., Gray, D., Read, K., Li, C., Parenteau, A., and Berthiaume, L.G. (2019). Initial Characterization and Toxicology of an Nmt Inhibitor in Development for Hematologic Malignancies. Blood 134, 3362-3362.

[0163] 6. Sangha, R., Mackey, J.R., Sehn, L.H., Kuruvilla, J., Weickert, M.J., and Berthiaume, L.G. (2021). An Open-Label, First-in-Human, Phase I Trial of Daily PCLX- 001. Blood 138, 1364-1364.

[0164] 7. Erwan Beauchamp, Chistopher Cromwell, Eman Moussa, Aishwarya Iyer, Megan Yap, Rony Pain, Jay Gamma, Olivier Julien, Basil Hubbard, Luc Berthiaume. Understanding the sensitivity of cancer cells to myristoylation inhibitors for oncology applications [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 1662.

[0165] 8. Erwan Beauchamp, Jay Gamma, Christopher R. Cromwell, Eman W. Moussa, Aishwarya Iyer, Megan Yap, Rony Pain, Morris Kostiuk, Krista M. Vincent, Lynne M. Postovit, Olivier Julien, Basil P. Hubbard, John R. Mackey & Luc G. Berthiaume. Myristoylation inhibition impacts oxidative phosphorylation and defines a 91 gene sensitivity signature that identifies new solid cancer indications. Manuscript submitted.

[0166] 8a AACR 2022: Jay Gamma, Erwan Beauchamp, Qiang Liu, Aishwarya Iyer, Megan Yap, Zoulika Zak, Cassidy Ekstrom, Krista Vincent, Joseph Brandwein, Jean C.Y.Wang, Naveen Pemmaraju, Gautam Borthakur, John R. Mackey, Luc G. Berthiaume; Targeting N-Myristoylation for Adult Acute Myeloid Leukemia Therapy. Blood 2022; 140 (Supplement 1): 088–3089. doi: https: / / doi.org / 10.1182 / blood-2022-170544 (abstract)

[0167] https: / / ashpublications.org / blood / article / 140 / Supplement%201 / 3088 / 490403 / T argeting-N-Myristoylation-for-Adult-Acute-Myeloid

[0168] 9. Zhang, Qingyu, Wei Zhou, Shan Yu, Yaojun Ju, Sally Kit Yan To, Alice Sze Tsai Wong, Yufei Jiao, Terence Chuen Wai Poon, Kin Yip Tam, and Leo Tsz On Lee. "Metabolic reprogramming of ovarian cancer involve ACSL1-mediated metastasis stimulation through upregulated protein myristoylation." Oncogene 40, no.1 (2021): 97-111)

[0169] 10. Rony Pain, Erwan Beauchamp, Katia Carmine-Simmen, Jay Gamma, Rebecca Reif, Abul Azad, Allan Murray, John Lewis, Luc Berthiaume; Abstract 3620: N- myristoylation inhibition reduces angiogenesis and cancer cell migration. Cancer Res 1 April 2023; 83 (7_Supplement): 3620. https: / / doi.org / 10.1158 / 1538-7445.AM2023-3620

[0170] The embodiments described herein are intended to be examples only. Alterations, modifications and variations can be effected to the particular embodiments by those of skill in the art. The scope of the claims should not be limited by the particular embodiments set forth herein, but should be construed in a manner consistent with the specification as a whole.

[0171] All publications, patents and patent applications mentioned in this Specification are indicative of the level of skill those skilled in the art to which this invention pertains and are herein incorporated by reference to the same extent as if each individual publication patent, or patent application was specifically and individually indicated to be incorporated by reference.

[0172] The invention being thus described, it will be obvious that the same may be varied in many ways. Such variations are not to be regarded as a departure from the spirit and scope of the invention, and all such modification as would be obvious to one skilled in the art are intended to be included within the scope of the following claims.

Claims

WHAT IS CLAIMED IS:

1. A method for treating cancer in a human subject in need thereof, comprising, administering an oral dosage form of PCLX-001, wherein said oral dosage comprises about 20mg PCLX-001 per day to about 210 mg PCLX-001 per day, wherein said oral dosage provides an effect comprising a Tmax of about 1 hours to about 4 hours or wherein said oral dosage provides an effect comprising a Tmax of about 1 hours to about 6 hours if said subject is taking a proton pump inhibitor (PPI).

2. The method of claim 1, wherein said oral dosage comprises about 20mg PCLX-001 per day, 40mg PCLX-001 per day, 70mg PCLX-001 per day, 100mg PCLX-001 per day, 140mg PCLX-001 per day, or 210mg PCLX-001 per day.

3. The method of claim 1 or 2, wherein said administration is daily administration from 1 to 28 days.

4. The method of any one of claims 1 to 3, wherein the cancer is ovarian cancer, refractory ovarian cancer, colorectal adenocarcinoma, or metastatic colorectal cancer.

5. The method of any one of claims 1 to 3, wherein the cancer is Diffuse Large B Cell Lymphoma (DLBCL).

6. The method of claim 5, wherein said DLBC is refractory DLBC.

7. The method of any one of claims 1 to 3, wherein the cancer is lymphoma, leukemia, myeloma, breast cancer, small cell lung cancer, non-small cell lung cancer cells; melanoma, adenocarcinoma, pancreas cancer, bladder cancer, ovarian cancer, brain cancer, colon cancer, rectal cancer, colorectal cancer, neuroblastoma, carcinoma, endometrial cancer, sarcoma, thyroid cancer, fibrosarcoma, oral cancer, tongue cancer, nasopharynx cancer, larynx cancer, esophageal cancer, germ cell cancer, stomach cancer, hepatocellular cancer,biliary tract cancer, small intestinal cancer, testis cancer, epidermoid cancer, cervical cancer, kidney cancer, prostate cancer, or pharynx cancer.

8. Use of an oral dosage of PCLX-001 for treating cancer in a human subject, wherein said oral dosage comprises about 20mg PCLX-001 per day to about 210 mg PCLX-001 per day, wherein said oral dosage provides an effect comprising a Tmax of about 1 hours to about 4 hours or wherein said oral dosage provides an effect comprising a Tmax of about 1 hours to about 6 hours if said subject is taking a proton pump inhibitor (PPI).

9. Use of an oral dosage of PCLX-001 in the manufacture of a medicament for treating cancer in a subject, wherein said oral dosage comprises about 20mg PCLX-001 per day to about 210 mg PCLX-001 per day, wherein said oral dosage provides an effect comprising a Tmax of about 1 hours to about 4 hours or wherein said oral dosage provides an effect comprising a Tmax of about 1 hours to about 6 hours if said subject is taking a proton pump inhibitor (PPI).

10. The use of claim 8 or 9, wherein said oral dosage comprises about 20mg PCLX-001 per day, 40mg PCLX-001 per day, 70mg PCLX-001 per day, 100mg PCLX-001 per day, 140mg PCLX-001 per day, or 210mg PCLX-001 per day.

11. The use of any one of claims8 to 10, wherein the cancer is ovarian cancer, refractory ovarian cancer, colorectal adenocarcinoma, or metastatic colorectal cancer.

12. The use of any one of claims 8 to 10, wherein the cancer is Diffuse Large B Cell Lymphoma (DLBCL).

13. The use of claim 12, wherein said DLBC is refractory DLBC.

14. The use of any one of claims 8 to 10, wherein the cancer is lymphoma, leukemia, myeloma, breast cancer, small cell lung cancer, non-small cell lung cancer cells; melanoma, adenocarcinoma, pancreas cancer, bladder cancer, ovarian cancer, brain cancer, coloncancer, rectal cancer, colorectal cancer, neuroblastoma, carcinoma, endometrial cancer, sarcoma, thyroid cancer, fibrosarcoma, oral cancer, tongue cancer, nasopharynx cancer, larynx cancer, esophageal cancer, germ cell cancer, stomach cancer, hepatocellular cancer, biliary tract cancer, small intestinal cancer, testis cancer, epidermoid cancer, cervical cancer, kidney cancer, prostate cancer, or pharynx cancer.

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