Hydroxylurea methyl acylflubene for the treatment of brain cancer or CNS cancer

Abstract

The present invention provides a pharmaceutical composition for treating brain cancer or central nervous system cancer. [Solution] A pharmaceutical composition is provided comprising hydroxyureamethyl-acylfluben or a pharmaceutically acceptable salt thereof as an active ingredient.

Description

Technical Field

[0001] The present application relates to the fields of chemistry and oncology. More specifically, the present application relates to a method of treating brain cancer using hydroxyureamethyl-acylfulvene. Background Art

[0002] Glioblastoma multiforme (GB, GBM, or glioblastoma) is ranked among the most aggressive human cancers, with a median survival of approximately 15 months. The current standard of care for GB consists of de-bulking surgery followed by combination therapy with fractionated ionizing radiation (IR) and temozolomide (TMZ), a DNA alkylating agent. Since the sensitivity of GBM to TMZ depends on the methylation status of O6-methylguanine methyltransferase (MGMT), a DNA repair enzyme, the effectiveness of standard treatment with TMZ is limited.

[0003] Glioblastoma is the most common and aggressive malignant brain tumor. Despite advances in treatment, improvements in overall survival remain limited. GB patients almost uniformly experience recurrence, and despite aggressive treatment with surgery, radiation, and chemotherapy, the median survival is only 15 to 20 months. Surgical resection of the entire tumor is extremely difficult and impossible in most cases. Glioblastoma has finger-like tentacles extending from the main tumor mass into surrounding normal brain tissue. In many cases, surgical resection is limited by the balance between tumor removal and the risk to cognitive function, or indeed between immediate patient survival. GB may proliferate or metastasize to other regions of the brain.

[0004] Another reason why GB is difficult to treat is that many drugs cannot efficiently cross the "blood-brain barrier" to enter the brain and act on the tumor. Glioblastoma contains heterogeneous cell populations, some of which respond to treatment and some of which do not. These non-responding cells survive, spread throughout the brain, and rarely achieve long-term success. These and other factors make GB a malignant tumor that is difficult to characterize and treat.

[0005] Therefore, there is always a need for improved methods and processes for treating GB and other brain cancers.

[0006] definition Unless otherwise defined, all technical and scientific terms used herein have the same meanings as those generally understood by those skilled in the art to which the subject matter of this specification belongs. For the purpose of facilitating the understanding of the invention, the following definitions are provided:

[0007] An amino acid sequence (referring to a mutant polypeptide) that aligns with the amino acid sequence described in Sequence ID X means that the mutant amino acid sequence and the amino acid sequence described in Sequence ID X are aligned in a preferred manner that enables sequence comparison and allows identification of the position in the mutant amino acid sequence where, when compared with the amino acid sequence described in Sequence ID X, the same amino acid is present (at the same position), a different amino acid is present (substitution), one or more extra amino acids are present (insertion or extension), or amino acids are absent (deletion or truncation).

[0008] As used herein, the term “antibody” includes intact molecules, as well as molecules containing or composed of fragments thereof, such as Fab, F(ab')2, Fv, and scFv, and engineered variants, including diabodies, triabodies, minibodies, and single-domain antibodies that can bind to epitope determinants. Thus, antibodies may exist as intact immunoglobulins or as various forms of modification.

[0009] As used herein, the terms “patient,” “subject,” “individual,” and “host” refer to either a human or non-human animal that is suffering from or suspected to be suffering from a disease or disorder associated with abnormal biological or cellular proliferation activity.

[0010]

[0024] The term "biomarker" refers to any molecule, such as a gene, gene transcript (e.g., mRNA), peptide or protein, or fragment thereof, produced by a subject that is useful in identifying a subject to predict a patient's response to a treatment containing 2,2'-dithio-bis-ethanesulfonate disodium or an analogue thereof. A biomarker that is differentially present (i.e., increased or decreased) in a biological sample from a subject or group of subjects is considered to have a first phenotype (e.g., disease-bearing) compared to a biological sample from a subject or group of subjects having a second phenotype (e.g., disease-free). Biomarkers can be differentially present at any level, but generally at at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 100%, at least 110%, at least 120%, at least 130%, and at least The biomarker is present at a level that increases by 140%, at least 150%, or more, or is generally present at a level that decreases by at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or 100% (i.e., absent). The biomarker is preferably present differentially at a statistically significant level (e.g., a p-value less than 0.05 and / or a q-value less than 0.10 determined using either Welch's t-test or Wilcoxon's rank-sum test).

[0011] The terms “glioblastoma multiforme,” “GB,” or “GBM” refer to the most common and aggressive type of primary brain tumor in humans. GB tumors are characterized by the presence of small areas of necrotic tissue surrounded by undifferentiated cells (pseudopalisade necrosis). This feature, as well as the presence of hyperplastic blood vessels, distinguishes this tumor from grade 3 astrocytomas, which lack these features. In some cases, the terms “glioblastoma multiforme,” “glioblastoma,” or “malignant glioma” are used interchangeably herein to refer to brain tumors arising from astrocytic cells.

[0012] The terms “to treat” and “to treat” in reference to such diseases or disorders refer to improving at least one symptom of the disease or disorder. When these terms are used in reference to conditions such as cancer, they refer to one or more of the following: inhibiting cancer growth, reducing the weight or volume of cancer, extending the patient’s expected survival, inhibiting tumor growth, reducing tumor burden, reducing the size or number of metastatic lesions, suppressing the development of new metastatic lesions, extending survival, extending progression-free survival, extending time to progression, and / or improving quality of life. The terms “to treat” or “to treat” glioblastoma multiforme may include preventing the development of glioblastoma, reversing symptoms, and / or improving the clinical outcome of a patient with glioblastoma or recurrent glioblastoma. Examples of improved clinical outcomes include extended survival, reduction in tumor size, non-proliferative tumors, and / or absence of worsening neurological symptoms. Non-specific examples of neurological symptoms include double vision, vomiting, loss of appetite, mood and personality changes, changes in thinking and learning abilities, seizures, speech disorders, and cognitive impairment.

[0013] When used in relation to a condition or disease such as cancer, the term “prevent” refers to a reduction in the frequency of symptoms of the condition or disease, or a delay in its onset. Therefore, cancer prevention includes, for example, reducing the number of detectable cancerous growths in a group of patients receiving prophylactic treatment compared to an untreated control group, and / or delaying the appearance of detectable cancerous growths in the treated group versus the untreated group by, for example, a statistically and / or clinically significant amount.

[0014] As used herein, the terms “expression level” and “expression level” refer to the amount of gene product, e.g., DNA, RNA (e.g., messenger RNA (mRNA)), or protein corresponding to a particular gene in a cell, tissue, biological specimen, organism, or patient.

[0015] The term "pharmaceutically acceptable" generally means safe, non-toxic, and useful for preparing a pharmaceutical composition that is biologically or otherwise desirable, and includes those that are acceptable for veterinary and human medicinal use.

[0016] "Pharmacologically acceptable salt" refers to a salt that is acceptable for administration to patients such as mammals (e.g., a salt that has mammalian safety acceptable for a given drug regimen). Such salts can be derived from pharmaceutically acceptable inorganic or organic bases and pharmaceutically acceptable inorganic or organic acids, depending on the specific substituents found in the compounds described herein. If the compounds of this disclosure contain relatively acidic functional groups, the base addition salt can be obtained by contacting the neutral form of such compound with a sufficient amount of the desired base in a pure (neat) or suitable inert solvent. Examples of salts derived from pharmaceutically acceptable inorganic bases include aluminum, ammonium, calcium, copper, iron, lithium, magnesium, manganese, potassium, sodium, and zinc. Examples of salts derived from pharmaceutically acceptable organic bases include salts of primary, secondary, tertiary, and quaternary amines, such as substituted amines, cyclic amines, and natural amines, including arginine, betaine, caffeine, choline, N,N'-dibenzylethylenediamine, diethylamine, 2-diethylaminoethanol, 2-dimethylaminoethanol, ethanolamine, ethylenediamine, N-ethylmorpholine, N-ethylpiperidine, glucamine, glucosamine, histidine, hydravamin, isopropylamine, lysine, methylglucamine, morpholine, piperazine, piperidine, polyamine resins, procaine, purine, theobromine, triethylamine, trimethylamine, tripropylamine, tromethamine, N,N'-dibenzylethylenediamine, chloroprocaine, choline, diethanolamine, meglumine (N-methylglucamine), and other substituted amines, cyclic amines, and natural amines. If the compounds of the present disclosure contain relatively basic functional groups, the acid addition salt can be obtained by contacting the neutral form of such compound with a sufficient amount of the desired acid in a pure or suitable inert solvent.Salts derived from pharmaceutically acceptable acids include acetate, trifluoroacetate, propionate, ascorbate, benzenesulfonate, benzoate, camphor sulfonate, citrate, ethanesulfonate, fumarate, glycolate, gluconate, glucuronate, glutamate, hippuric acid, hydrobromide, hydrochloride, isethionate, lactate, lactobionate, maleate, malate, mandelate, methanesulfonate, mucoate, naphthalenesulfonate, nicotinate, nitrate, pamoate, pantothenate, and phosphate. Examples include salts, succinates, sulfates, hydroiodides, carbonates, tartrates, p-toluenesulfonates, pyruvates, aspartates, benzoates, anthranilates, mesylates, salicylates, p-hydroxybenzoates, phenylacetates, embonic (pamoic), ethanesulfonates, benzenesulfonates, 2-hydroxyethanesulfonates, sulfanilates, stearates, cyclohexylaminosulfonates, algenic salts, hydroxybutyrates, galactarates, and galacturonates.

[0017] The term "therapeutic effect" refers to a beneficial local or systemic effect in animals, particularly mammals, and more specifically humans, caused by the administration of the compound or composition of the present invention. The term "therapeutic effective dose" means the amount of the compound or composition of the present invention that is effective in treating a disease or condition caused by abnormal biological activity in a reasonable benefit / risk ratio. In some embodiments, the therapeutically effective dose of hydroxyureamethylacylfluben or a pharmaceutically acceptable salt thereof is selected from the group consisting of 0.5 mg / day, 1 mg / day, 2.5 mg / day, 5 mg / day, 10 mg / day, 20 mg / day, 30 mg / day, 60 mg / day, 90 mg / day, 120 mg / day, 150 mg / day, 180 mg / day, 210 mg / day, 240 mg / day, 270 mg / day, 300 mg / day, 360 mg / day, 400 mg / day, 440 mg / day, 480 mg / day, 520 mg / day, 580 mg / day, 600 mg / day, 620 mg / day, 640 mg / day, 680 mg / day, and 720 mg / day.

[0018] The term “healthy individual” is interpreted to mean an individual known not to have cancer, and such knowledge is derived from clinical data of the individual, including but not limited to diagnostic assays other than those described herein.

[0019] "Reference level" means the level of the compound or additional biomarker of the present invention that indicates a specific disease state, phenotype, or lack thereof, or a combination of disease states, phenotypes, or lack thereof.

[0020] A "reference sample" refers to a sample containing a reference level of biomarkers. For example, reference samples can be obtained from subjects that do not have a specific disease, medical condition, or phenotype such as cancer or acute injury.

[0021] The therapeutically effective dose of such a substance varies depending on the subject and the condition being treated, the subject's weight and age, the severity of the condition, the method of administration, etc., and these can be easily determined by those skilled in the art. [Overview of the project]

[0022] One aspect of this application includes a method for therapeutically treating brain cancer, comprising administering an effective amount of hydroxyureamethyl-acylfluben to a subject in need thereof. In one embodiment, the brain cancer is glioblastoma multiforme.

[0023] Another aspect of this application includes a method for treating brain cancer, comprising using an effective amount of hydroxyureamethyl acylfluben together with an effective amount of one or more additional therapeutic agents selected from the group consisting of temozolomide, bevacizumab, everolimus, carmustine, lomustine, procarbazine, vincristine, irinotecan, cisplatin, carboplatin, methotrexate, etoposide, vinblastine, bleomycin, actinomycin, cyclophosphamide, and ifosfamide.

[0024] Another aspect of the present application includes a method for treating brain cancer by using an effective amount of hydroxyureamethyl-acylfulvene in combination with radiation therapy. The radiation therapy can be selected from whole brain irradiation, fractionated radiation therapy, radiosurgery, and combinations thereof.

[0025] Another aspect of the present application includes the method wherein the patient is a human or an animal. BRIEF DESCRIPTION OF THE DRAWINGS

[0026] [Figure 1] It shows that hydroxyureamethyl-acylfulvene exhibits nanomolar potency in GB cell lines. [Figure 2] It shows that hydroxyureamethyl-acylfulvene exhibits nanomolar potency in additional GB cell lines. [Figure 3] It shows an analysis of the modulation of the blood-brain barrier by hydroxyureamethyl-acylfulvene. [Figure 4] It shows the effect on cell viability in the presence of hydroxyureamethyl-acylfulvene. [Figure 5] It shows a profile of in vivo hydroxyureamethyl-acylfulvene activity in the GBM U87 subcutaneous (Sc) xenograft tumor model. [Figure 6A] It shows the results of treatment with vehicle as a control in mice transplanted with GBM neurospheres (M1123). [Figure 6B] It shows the results of treatment with 4 mg / kg of hydroxyureamethyl-acylfulvene administered intravenously four times every other day in mice transplanted with GBM neurospheres (M1123). [Figure 7] It shows images of physical observation of mice transplanted with GBM neurospheres (M1123) treated with a vehicle control or hydroxyureamethyl-acylfulvene. MODES FOR CARRYING OUT THE INVENTION

[0027] Sequence SEQ ID NO: 1 -- the amino acid sequence of EGFR,

[0028] The amino acid sequence of sequence number 2-NF1,

[0029] SEQ ID NO: 3 -- Amino acid sequence of IDH1,

[0030] SEQ ID NO: 4 -- Amino acid sequence of LAB1,

[0031] SEQ ID NO: 5 -- Amino acid sequence of IGDH,

[0032] SEQ ID NO: 6 -- Amino acid sequence of ANXA2,

[0033] The amino acid sequence of sequence number 7--S100A11,

[0034] SEQ ID NO: 8 -- Amino acid sequence of CTSB,

[0035] The amino acid sequence of sequence number 9-TP53, and

[0036] Sequence ID 10 -- Amino acid sequence of MGMT

[0037] Hydroxyureamethylacylfluben or hydroxyureamethyl-acylfluben (currently referred to as LP-184 by Lantern Pharma, Inc.) is a semi-synthetic or synthetic antitumor drug derived from the mushroom toxin illudin S. The structures of each isomer are shown below. [ka]

[0038] Certain embodiments relate to a method for treating brain cancer, the method comprising administering an effective amount of hydroxyureamethyl-acylfluben to a subject in need thereof. In some embodiments, the brain cancer may originate from metastatic brain tumors, glioblastoma multiforme, or a combination thereof. In a particular example, the brain tumor is glioblastoma multiforme. In one example, hydroxyureamethyl-acylfluben may be administered as monotherapy.

[0039] One embodiment involves administering hydroxyureamethyl-acylfluben and an additional therapeutic agent in separate compositions or simultaneously in the same composition. Therefore, some embodiments include a first pharmaceutical composition comprising (a) a safe and therapeutically effective amount of hydroxyureamethyl-acylfluben or a pharmaceutically acceptable salt thereof, and (b) a pharmaceutically acceptable carrier, diluent, excipient, or combination thereof, and a second pharmaceutical composition comprising (1) a safe and therapeutically effective amount of an additional therapeutic agent, and (2) a pharmaceutically acceptable carrier, diluent, excipient, or combination thereof. Some embodiments include a pharmaceutical composition comprising (a) a safe and therapeutically effective amount of an additional therapeutic agent, and (b) a pharmaceutically acceptable carrier, diluent, excipient, or combination thereof. In some embodiments, the methods described herein may further include subjecting the subject to radiotherapy. In some embodiments, the radiotherapy may be whole-brain irradiation, fractional radiotherapy, and radiosurgery.

[0040] Another embodiment includes a method for inhibiting the proliferation of brain tumor cells, which involves contacting brain tumor cells with hydroxyureamethyl-acylfluben. In some embodiments, contact involves administering an effective amount of hydroxyureamethyl-acylfluben to a subject having brain tumor cells. In some embodiments, the brain tumor is glioblastoma multiforme. In some embodiments, this method can be used to treat primary central nervous system (CNS) tumors. CNS cancers are a type of cancer that occurs in the central nervous system. These include brainstem gliomas, craniopharyngiomas, medulloblastomas, and meningiomas. CNS tumors arise from normal cells of the brain and spinal cord called “neurons” and “glia.” Tumors arising from neurons include medulloblastomas and primitive neuroectodermal tumors (PNETs). Tumors arising from glia include gliomas, astrocytomas, oligodendrogliomas, and ependymoblastomas. The specific names of tumors often reflect the tissue of origin of the CNS tumor.

[0041] Another embodiment is a method for therapeutically treating brain cancer or CNS tumors in a subject, comprising the steps of: obtaining a tumor sample from the subject; measuring the expression of MGMT+ or MGMT- in the tumor sample; and, if MGMT+ is measured to be higher than a baseline, administering an effective amount of hydroxyureamethyl-acylfluben or a pharmaceutically acceptable salt thereof to the subject in need, wherein the baseline is the level of MGMT in a healthy person. In one example, the characterization further includes molecular subtype classification as classical, mesenchymal, proneural, or neural glioblastoma.

[0042] The protein sequences of the specific proteins identified herein include EGFR (SEQ ID NO: 1), NF1 (SEQ ID NO: 2), IDH1 (SEQ ID NO: 3), LAMB1 (SEQ ID NO: 4), UGDH (SEQ ID NO: 5), ANXA2 (SEQ ID NO: 6), S100A11 (SEQ ID NO: 7), CTSB (SEQ ID NO: 8), TP53 (SEQ ID NO: 9), and MGMT (SEQ ID NO: 10).

[0043] Some embodiments include a method for inducing apoptosis in brain tumor cells, the method comprising contacting brain tumor cells with hydroxyureamethyl-acylfluben. In some embodiments, contact comprises administering an effective amount of hydroxyureamethyl-acylfluben to a subject having brain tumor cells. In some embodiments, the brain tumor is glioblastoma multiforme.

[0044] Some embodiments include a treatment for GB, comprising the steps of (1) performing a physical examination of the patient; (2) using magnetic resonance imaging ("MRI") to confirm the presence of a tumor in the patient's brain; (3) performing a brain biopsy to obtain genetic information about the cancer, including molecular subtypes and genetic markers; and (4) prescribing an effective treatment of hydroxyureamethylacylflubene to a subject having GB.

[0045] The duration of administration can be a treatment cycle of several weeks, as long as the tumor is under control and the regimen is clinically tolerable. In some embodiments, a single dose of hydroxyureamethyl-acylfluben or other therapeutic agent may be administered once a week, preferably once on day 1 and once on day 8 of a 3-week (21-day) treatment cycle. In some embodiments, a single dose of hydroxyureamethyl-acylfluben or other therapeutic agent may be administered once a week, twice a week, three times a week, four times a week, five times a week, six times a week, or daily during a 1-week, 2-week, 3-week, 4-week, or 5-week treatment cycle. Administration may be on the same day or different days each week of the treatment cycle.

[0046] Another embodiment includes a method for determining the treatment or sensitivity of brain cancer (e.g., glioblastoma or CNS cancer) to hydroxyureamethyl acylfluben therapy by evaluating the level of MGMT expression. Higher levels of MGMT than in healthy individuals indicate efficacy of LP-184.

[0047] A method for treating brain cancer or CNS cancer, comprising the steps of: detecting the presence of specific genetic information in a human subject; and, if the human subject overexpresses or underexpresses a specific marker, administering hydroxyureamethylacylfluben or a pharmaceutically acceptable salt thereof to the subject. The marker may be substantially similar to EGFR (SEQ ID NO: 1), NF1 (SEQ ID NO: 2), IDH1 (SEQ ID NO: 3), LAMB1 (SEQ ID NO: 4), UGDH (SEQ ID NO: 5), ANXA2 (SEQ ID NO: 6), S100A11 (SEQ ID NO: 7), CTSB (SEQ ID NO: 8), TP53 (SEQ ID NO: 9), and MGMT (SEQ ID NO: 10).

[0048] Another embodiment includes a method for treating or determining the sensitivity of brain cancer (e.g., glioblastoma) to hydroxyurea methyl acylflubene therapy by evaluating the level of MGMT expression and / or at least one gene selected from the group consisting of laminin subunit beta-1 (LAMB1), UDP-glucose 6-dehydrogenase (UGDH), annexin A2 (ANXA2), S100 calcium-binding protein A11 (S100A11), and cathepsin B (CTSB). The nucleotide and / or amino acid sequences are well known and published in public databases (e.g., Genebank) and available publications.

[0049] Another embodiment is a method for treating brain cancer in a subject, comprising the steps of (a) obtaining or having obtained expression levels in a sample from the subject for a plurality of targets, wherein the plurality of targets include the group consisting of MGMT (SEQ ID NO: 10), LAMB1 (SEQ ID NO: 4), UGDH (SEQ ID NO: 5), ANXA2 (SEQ ID NO: 6), S100A11 (SEQ ID NO: 7), and / or CTSB (SEQ ID NO: 8); (b) determining that the subject is sensitive to treatment with hydroxyurea methyl acylflubene; and (c) administering a cancer treatment comprising hydroxyurea methyl acylflubene.

[0050] Another embodiment is a method for treating brain cancer in a subject, comprising the steps of (a) obtaining or having obtained expression levels of a plurality of targets in a sample from the subject, wherein the plurality of targets include the group consisting of EGFR, NF1, and / or PDGFRA / IDH1; (b) determining that the subject is sensitive to treatment with hydroxyurea methyl acylflubene; and (c) administering a cancer treatment comprising hydroxyurea methyl acylflubene.

[0051] Another embodiment includes the detection of genetic information, including high levels of EGFR amplification, deletion of the NF1 gene, high expression of the MET gene, alteration of the PDFRA gene, point mutations in IDH1, TP53 mutations, expression of neuronal markers by astrocytic cells, mutations in oncogenes, MGMT methylation, or combinations thereof.

[0052] Hydroxyurea methylacylfluben for use in accordance with the present invention can be administered primarily by parenteral administration, specifically including subcutaneous, intramuscular, intravenous, transdermal, intrathecal, epidural, intra-articular, and topical administration, or by oral administration if possible, in various dosage forms.

[0053] Examples of injectable preparations for parenteral administration include sterile aqueous or non-aqueous solutions, suspensions, and emulsions. Examples of aqueous solutions and suspensions include distilled water for injection and physiological saline. Examples of non-aqueous solutions and suspensions include propylene glycol, polyethylene glycol, vegetable oils such as olive oil, alcohols such as ethanol, and polysorbate 80 (trade name). Such compositions may contain auxiliary agents such as preservatives, wetting agents, emulsifiers, dispersants, stabilizers (e.g., lactose), and solubilizers (e.g., meglumine). These are sterilized by filtration with a bacteria-retaining filter, formulation with a bactericide, or irradiation. Alternatively, they may be prepared once as a sterile solid composition and then dissolved or suspended in sterile water or a sterile solvent for injection before use.

[0054] In some embodiments, the brain tumor can be selected from metastatic brain tumors, anaplastic astrocytomas, glioblastomas multiforme, oligodendrogliomas, ependymomas, meningiomas, mixed gliomas, and combinations thereof. In some embodiments, the brain tumor is a glioblastoma multiforme. In some embodiments, the brain tumor is a metastatic brain tumor.

[0055] In some embodiments, the brain tumor can be selected from anaplastic astrocytoma, central neurocytoma, choroid plexus carcinoma, choroid plexus papilloma, choroid plexus tumor, germinal dysplastic neuroepithelial tumor, ependymal tumor, fibrous astrocytoma, giant cell glioblastoma, multiform glioblastoma, cerebral glioma, gliosarcoma, perivascular cell tumor, medulloblastoma, medullary epithelioma, meningeal carcinomatosis, neuroblastoma, neurocytoma, oligodendroglioma, oligodendroglioma, optic nerve sheath meningioma, pediatric ependymoma, pilocytic astrocytoma, pineoblastoma, pineocytoma, pleomorphic anaplastic neuroblastoma, pleomorphic xanthoblastoma, primary central nervous system lymphoma, butterfly tumor, subependymal giant cell astrocytoma, subependymal tumor, central nervous system myeloma, and trilateral retinoblastoma.

[0056] Liquid compositions for oral administration include, for example, pharmaceutically acceptable emulsions, liquids, suspensions, syrups, and elixirs, and include, for example, distilled water and ethanol as inert diluents for general use. In addition to inert diluents, the compositions may also contain auxiliary agents such as wetting agents and suspending agents, sweeteners, flavoring agents, fragrances, and preservatives.

[0057] The active compound can also be administered intravenously or intraperitoneally by injection or infusion. Solutions of the active compound can be prepared in water and may be mixed with non-toxic surfactants. Dispersions can also be prepared in glycerol, liquid polyethylene glycol, triacetin, and mixtures thereof, and in oil. Under normal storage and use conditions, these formulations contain preservatives to prevent microbial growth.

[0058] It should be understood that specific dosages and treatment plans for any particular patient will depend on a variety of factors, including the activity of the specific compound used, age, weight, overall health, sex, diet, administration time, excretion rate, drug combinations, the judgment of the treating physician, and the severity of the specific disease being treated. The amount of the compound of the present invention in the composition will also depend on the specific compound in the composition. [Examples]

[0059] Example 1 Table 1 shows that LP-184 / hydroxyureamethyl-acylfluben exhibits high sensitivity in tumors expressing MGMT. In addition to MGMT expression, other GB-specific genes also show multi-omics parameter correlations with the possibility of LP-184 / hydroxyureamethyl-acylfluben being sensitive to GB.

[0060] Table 1 shows genes that can be used to indicate GB sensitivity to hydroxyureamethyl-acylflubene. [Table 1]

[0061] Table 2 shows genes associated with TMZ resistance that are downregulated in GB and negatively correlated with LP-184 sensitivity; therefore, LP-184 may be beneficial in such tumors. [Table 2]

[0062] Table 3 shows genes whose upregulation is known to promote GB. Tumors with elevated expression of such genes (compared to healthy individuals) may be more susceptible to hydroxyureamethylacylflubene. [Table 3]

[0063] Example 2 Figure 1 shows that hydroxyureamethyl acylflubene exhibits nanomolar potency in GB cell lines. The cytotoxicity of LP-184 in terms of IC50 in four cell lines ranges from 45 to 230 nM.

[0064] Results from tools used to evaluate chemical ADMET (absorption, distribution, metabolism, excretion-toxicity) properties indicate that LP-184 crosses the blood-brain barrier. NC160 data showed that drug sensitivity-gene expression correlation analysis of hydroxyureamethyl-acylfluben and the TMZ precursor molecule mitozolomide did not reveal any duplicate genes likely to mediate resistance to either drug. Hydroxyureamethyl-acylfluben may be useful as a potential monotherapy or combination therapy for GB tumors. Example 3

[0065] Hydroxyureamethyl-acylfluben was tested and showed activity in a broad spectrum of in vitro and in vivo brain cancer models representing various molecular and clinical subtypes of glioblastoma. Hydroxyureamethyl-acylfluben was shown to cross the blood-brain barrier in the in vitro model system.

[0066] Figure 2 shows that hydroxyureamethyl acylflubene exhibits nanomolar potency in various GB cell lines beyond that shown in Figure 1. The cytotoxicity of hydroxyureamethyl acylflubene in terms of IC50 in a series of six cell lines ranged from 46 to 2620 nM.

[0067] Figure 3 shows an analysis of the regulation of the blood-brain barrier by compound interactions. Hydroxyureamethyl-acylfluben did not significantly affect cell viability, and the integrity of the blood-brain barrier was not impaired. To evaluate the in vivo therapeutic activity of hydroxyureamethyl-acylfluben in mice with glioblastoma, U87-SC cells (without in vitro drug treatment) were subcutaneously inoculated.

[0068] The physical, metabolic, and immunological privileged status of the brain presents diagnostic and therapeutic limitations, along with treatment-related neurotoxicity issues. A key characteristic of drugs against GBM is their ability to cross the blood-brain barrier (BBB). An independent in vitro study evaluating the cross-barrier kinetics of hydroxyureamethyl-acylflubene demonstrated that it crosses the blood-brain barrier. In a highly representative neuromics in vitro 3D model that closely replicates the human blood-brain barrier, the transport properties of the blood-brain barrier are reflected due to junctional zone formation, higher expression of specific carriers, or superior cell viability. The blood-brain barrier in this 3D in vitro model was created by co-culturing endothelial cells with pericytes and astrocytes in layers on an insert, improving endothelial cell polarization, promoting junctional zone formation, providing better interendothelial cell contact crucial for barrier development, and preventing dilution of secreted neurotrophic factors. These conditions combined lead to the development of an in vivo model that can truly mimic the blood-brain barrier. This assay utilizes a novel 3D blood-brain barrier model that allows for the study of both the effects of compounds on the structure and function of the blood-brain barrier, as well as the transport of compounds across the barrier. The behavior of LP-184 and TMZ, along with the current standard of treatment chemotherapy for GBM, was compared using known positive and negative controls, antipyrine and cyclosporine A, as benchmarks. LP-184 is as effective as TMZ in crossing the blood-brain barrier. As shown in the following graph, the apparent blood-brain barrier permeability measured for TMZ was 1.72*10⁴ cm / sec at 30 minutes, while for LP-184 it was 1.53*10⁴ cm / sec at 30 minutes.

[0069] Figure 4 shows the effect on cell viability after treatment with hydroxyureamethyl-acylfluben.

[0070] Figure 5 shows the in vivo hydroxyureamethylacylfluben activity profile in the GB U87-Sc xenograft tumor model. Mice in the U87-SC model were treated with hydroxyureamethylacylfluben at a dose of 4 mg / kg intravenously every other day for four doses. As a result, all five treated mice showed complete tumor regression, with no measurable tumors 12 days after the last dose.

[0071] Figures 6A and 6B show that mice (M1123) treated with intravenous administration of 4 mg / kg of hydroxyurea methyl-acylfluben every other day for four doses resulted in complete tumor regression. Figure 6A shows the results from mice (controls) transplanted with vehicle-treated GBM neurospheres (M1123). Figure 6B shows the results of treating mice transplanted with GBM neurospheres (M1123) with intravenous administration of 4 mg / kg of hydroxyurea methyl-acylfluben every other day for four doses. Three out of four mice showed no measurable tumors 12 days after the last dose.

[0072] Figure 7 shows images of physical observations of mice transplanted with GBM neurospheres (M1123) either as a vehicle control or treated with hydroxyureamethyl-acylfluben. Tumor volume showed clear inhibition of tumor growth in hydroxyureamethyl-acylfluben-treated mice compared to vehicle-treated control mice. This example demonstrates the in vivo efficacy of hydroxyureamethyl-acylfluben in a subcutaneous tumor model of GB.

[0073] Protein sequences of EGFR (SEQ ID NO: 1), NF1 (SEQ ID NO: 2), IDH1 (SEQ ID NO: 3), LAMB1 (SEQ ID NO: 4), UGDH (SEQ ID NO: 5), ANXA2 (SEQ ID NO: 6), S100A11 (SEQ ID NO: 7), CTSB (SEQ ID NO: 8), TP53 (SEQ ID NO: 9), and MGMT (SEQ ID NO: 10) from the Uniprot database: Sequence ID 1 (Sequence ID 1) EGFR / Homo sapiens / Human Sequence ID 2 NF1 / Homo sapiens / Human Sequence ID 3 (Sequence ID 3) IDH1 / Homo sapiens / Human MSKKISGGSVVEMQGDEMTRIIWELIKEKLIFPYVELDLHSYDLGIENRDATNDQVTKDAAEAIKKHNVGVKCATITPDEKRVEEFKLKQMWKSPNGTIRNIL GGTVFREAIICKNIPRLVSGWVKPIIIGRHAYGDQYRATDFVVPPGGKVEITYTPSDGTQKVTYLVHNFEEGGGVAMGMYNQDKSIEDFAHSSFQMALSKGWPL YLSTKNTILKKYDGRFKDIFQEIYDKQYKSQFEAQKIWYEHRLIDDMVAQAMKSEGGFIWACKNYDGDVQSDSVAQGYGSLGMMTSVLVCPDGKTVEAEAAHG TVTRHYRMYQKGQETSTNPIASIFAWTRGLAHRAKLDNNKELAFFANALEEVSIETIEAGFMTKDLAACIKGLPNVQRSDYLNTFEFMDKLGENLKIKLAQAKL Sequence ID 4 (Sequence ID 4) LAMB1 / Homo sapiens / Human MGLLQLLAFSFLALCRARVRAQEPEFSYGCAEGSCYPATGDLLIGRAQKLSVTSTCGLHKPEPYCIVSHLQEDKKCFICNSQDPYHETLNPDSHLIENVVTTFAPNRLKIWWQSENGVEN VTIQLDLEAEFHFTHLIMTFKTFRPAAMLIERSSDFGKTWGVYRYFAYDCEASFPGISTGPMKKVDDIICDSRYSDIEPSTEGEVIFRALDPAFKIEDPYSPRIQNLLKITNLRIKFVKL Sequence ID 5 UGDH / Homo sapiens / Human MFEIKKICCIGAGYVGGPTCSVIAHMCPEIRVTVVDVNESRINAWNSPTLPIYEPGLKEVVESCRGKNLFFSTNIDDAIKEADLVFISVNTPTKTYGMGKGRAADLKYIEACARRIVQNSNGY KIVTEKSTVPVRAAESIRRIFDANTKPNLNLQVLSNPEFLAEGTAIKDLKNPDRVLIGGDETPEGQRAVQALCAVYEHWVPREKILTTNTWSSELSKLAANAFLAQRISSINSISALCEATGAD VEEVATAIGMDQRIGNKFLKASVGFGGSCFQKDVLNLVYLCEALNLPEVARYWQQVIDMNDYQRRRFASRIIDSLFNTVTDKKIAILGFAFKKDTGDTRESSSIYISKYLMDEGAHLHIYDPK VPREQIVVDLSHPGVSEDDQVSRLVTISKDPYEACDGAHAVVICTEWDMFKELDYERIHKKMLKPAFIFDGRRVLDGLHNELQTIGFQIETIGKKVSSKRIPYAPSGEIPKFSLQDPPNKKPKV Sequence ID 6 (Sequence ID 6) ANXA2 / Homo sapiens / Human MSTVHEILCKLSLEGDHSTPPSAYGSVKAYTNFDAERDALNIETAIKTKGVDEVTIVNILTNRSNAQRQDIAFAYQRRTKKELASALKSALSGHLETVILGLLKTPAQYDASELKASMKGLGTDEDSLIEIICSRTNQELQEINRVYKEMYKTDLEKDIISDTSGDFRK LMVALAKGRRAEDGSVIDYELIDQDARDLYDAGVKRKGTDVPKWISIMTERSVPHLQKVFDRYKSYSPYDMLESIRKEVKGDLENAFLNLVQCIQNKPLYFADRLYDSMKGKGTRDKVLIRIMVSRSEVDMLKIRSEFKRKYGKSLYYYIQQDTKGDYQKALLYLCGGDD Sequence ID 7 (Sequence ID 7) S100A11 / Homo sapiens / Human MAKISSPTETERCIESLIAVFQKYAGKDGYNYTLSKTEFLSFMNTELAAFTKNQKDPGVLDRMMKKLDTNSDGQLDFSEFLNLIGGLAMACHDSFLKAVPSQKRT Sequence ID 8 (Sequence ID 8) CTSB / Homo sapiens / Human MWQLWASLCCLLVLANARSRPSFHPLSDELVNYVNKRNTTWQAGHNFYNVDMSYLKRLCGTFLGGPKPPQRVMFTEDLKLPASFDAREQWPQCPTIKEIRDQGSCGSCWAFGAVEAISDRICIHTNAHVSVEVSAEDLLTCCGSMCGDGCNGGYPAEAWNFWTRKGLVS GGLYESHVGCRPYSIPPCEHHVNGSRPPCTGEGDTPKCSKICEPGYSPTYKQDKHYGYNSYSVSNSEKDIMAEIYKNGPVEGAFSVYSDFLLYKSGVYQHVTGEMMGGHAIRILGWGVENGTPYWLVANSWNTDWGDNGFFKILRGQDHCGIESEVVAGIPRTDQYWEKI Sequence ID 9 (Sequence ID 9) TP53 / Homo sapiens / Human MEEPQSDPSVEPPLSQETFSDLWKLLPENNVLSPLPSQAMDDLMLSPDDIEQWFTEDPGPDEAPRMPEAAPPVAPAPAAPTPAAPAPAPSWPLSSSVPSQKTYQGSYGFRLGFLHSGTAKSVTCTYSPALNKMFCQLAKTCPVQLWVDSTPPPGTRVRAMAIYKQSQHMTEVVRRCPHHERCSDSDGLAPPQHLIR VEGNLRVEYLDDRNTFRHSVVVPYEPPEVGSDCTTIHYNYMCNSSCMGGMNRRPILTIITLEDSSGNLLGRNSFEVRVVCACPGRDRRTEEENLRKKGEPHHELPPGSTKRALPNNTSSSPQPKKKPLDGEYFTLQIRGRERFEMFRELNEALELKDAQAGKEPGGSRAHSSHLKKSKKGQSTSRHKKLMFKTEGPDSD Sequence ID 10 (Sequence ID 10) MGMT / Homo sapiens / Human MDKDCEMKRTTLDSPLGKLELSGCEQGLHEIKLLGKGTSAADAVEVPAPAAVLGGPEPLMQCTAWLNAYFHQPEAIEEFPVPALHHPVFQQESFTRQVLWKLL KVVKFGEVISYQQLAALAGNPKAARAVGGAMRGNPVPILIPCHRVVCSSGAVGNYSGGLAVKEWLLAHEGHRLGKPGLGGSSGLAGAWLKGAGATSGSPPAGRN

Claims

1. A method for therapeutically treating brain cancer or CNS cancer, comprising administering an effective amount of hydroxyureamethylacylfluben to a subject in need thereof.

2. The method according to claim 1, wherein the brain cancer is glioblastoma multiforme.

3. The method according to claim 1, wherein the additional therapeutic agent is selected from the group consisting of temozolomide, bevacizumab, everolimus, carmustine, lomustine, procarbazine, vincristine, irinotecan, cisplatin, carboplatin, methotrexate, etoposide, vinblastine, bleomycin, actinomycin, cyclophosphamide, and ifosfamide.

4. The method according to the claim, wherein the additional therapeutic agent is selected from the group consisting of cisplatin, paclitaxel, and other available therapies.

5. The method according to claim 1, further comprising subjecting the subject to radiotherapy.

6. The method according to claim 1, wherein the radiotherapy is selected from whole-brain irradiation, fractionated radiotherapy, radiosurgery, and combinations thereof.

7. The method according to claim 1, wherein the cells are located within an animal.

8. The method according to claim 1, wherein the subject or mammal is human.

9. The method according to claim 1, further comprising measuring the expression of brain cancer gene information and determining whether the expression of the gene information is greater than or less than a reference level for the gene information.

10. The method according to claim 8, wherein the gene information is EGFR, NF1, PDGFRA, or IDH1.

11. The method according to claim 8, wherein the gene information is LAMB1, UGDH, ANXA2, S100A11, and CTSB.

12. The method according to claim 8, wherein the genetic information is high-level EGFR amplification, deletion of the NF1 gene, high expression of the MET gene, alteration of the PDFRA gene, point mutation of IDH1, TP53 mutation, expression of neuronal markers by astrocytocytes, mutation of oncogenes, MGMT methylation, or a combination thereof.

13. The method according to claim 1, further comprising measuring the expression of MGMT+ or MGMT-.

14. The method according to claim 14, wherein the characterization further includes molecular subtype classification as classical, mesenchymal, proneuroral, or neural glioblastoma.

15. A method for therapeutically treating brain cancer or CNS cancer in a subject, The steps include obtaining a tumor sample from the aforementioned subject, The steps include measuring the expression of MGMT+ or MGMT- in the tumor sample, A method comprising the step of administering an effective amount of hydroxyureamethyl-acylfluben or a pharmaceutically acceptable salt thereof to a subject in need, where the MGMT is measured to be higher than a standard, the standard being the level of MGMT in a healthy person.

16. The method according to claim 15, further comprising a molecular subtype classification of the brain cancer as classical, mesenchymal, anterior nerve, or glioblastoma.

17. A method of treating brain cancer, a) A step of detecting the presence of genetic information in a human subject, b) The step of administering hydroxyureamethyl-acylfluben or a pharmaceutically acceptable salt thereof to the subject, c) The step of administering the hydroxyureamethyl-acylfluben or a pharmaceutically acceptable salt thereof, and then obtaining a tumor sample from the subject, d) Measuring the level of the marker in the tumor sample compared to a reference level, wherein the reference level is the MGMT level of a healthy person; e) A method comprising the step of continuing to administer the hydroxyureamethyl-acylfluben or a pharmaceutically acceptable salt thereof to the subject if a mutation is detected.

18. The method according to claim 17, wherein the marker is substantially similar to EGFR (SEQ ID NO: 1), NF1 (SEQ ID NO: 2), IDH1 (SEQ ID NO: 3), LAMB1 (SEQ ID NO: 4), UGDH (SEQ ID NO: 5), ANXA2 (SEQ ID NO: 6), S100A11 (SEQ ID NO: 7), CTSB (SEQ ID NO: 8), TP53 (SEQ ID NO: 9), and MGMT (SEQ ID NO: 10).

Images