WO2023113538A1 - Polythérapie à base de griséofulvine pour le traitement d'une tumeur cérébrale - Google Patents

Polythérapie à base de griséofulvine pour le traitement d'une tumeur cérébrale Download PDF

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Publication number
WO2023113538A1
WO2023113538A1 PCT/KR2022/020588 KR2022020588W WO2023113538A1 WO 2023113538 A1 WO2023113538 A1 WO 2023113538A1 KR 2022020588 W KR2022020588 W KR 2022020588W WO 2023113538 A1 WO2023113538 A1 WO 2023113538A1
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WIPO (PCT)
Prior art keywords
protein
temozolomide
pcnt
pericentrin
griseofulvin
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PCT/KR2022/020588
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English (en)
Korean (ko)
Inventor
장창영
백선하
박순범
홍지희
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숙명여자대학교산학협력단
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Priority claimed from KR1020220175813A external-priority patent/KR20230092793A/ko
Application filed by 숙명여자대학교산학협력단 filed Critical 숙명여자대학교산학협력단
Publication of WO2023113538A1 publication Critical patent/WO2023113538A1/fr

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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/335—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin
    • A61K31/34—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin having five-membered rings with one oxygen as the only ring hetero atom, e.g. isosorbide
    • A61K31/343—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin having five-membered rings with one oxygen as the only ring hetero atom, e.g. isosorbide condensed with a carbocyclic ring, e.g. coumaran, bufuralol, befunolol, clobenfurol, amiodarone
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/495—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with two or more nitrogen atoms as the only ring heteroatoms, e.g. piperazine or tetrazines
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents

Definitions

  • the present invention provides glyceofulvin combination therapy for the treatment of brain tumors.
  • Glioblastoma Multiforme is a type of brain tumor arising from glial cells and is a malignant tumor with the highest fatality rate occurring in the brain.
  • radiotherapy and temozolomide an oral treatment, are used as standard treatments for brain tumors, and the treatment effect is insignificant due to drug resistance.
  • Temozolomide is an anticancer drug that attaches a methyl group to DNA. Damaged DNA such as N7-meG or O6-meG cannot be repaired, causing double strand breaks and eventually causing apoptosis, thereby killing cancer cells.
  • MGMT O(6)-methylguanine-DNA methyltransferase
  • Glioblastoma which accounts for about 12-15% of all brain tumors, has an average survival time of only 14 months even after surgery, radiation, and chemotherapy, so the development of new treatments is urgent.
  • An object of the present invention is to provide a pharmaceutical composition for treating or preventing brain tumors containing griseofulvin and temozolomide.
  • Another object of the present invention is to provide a biomarker composition for predicting brain tumor prognosis comprising PCNT (pericentrin) protein or a gene encoding the protein as an active ingredient.
  • PCNT peripheral neuronucleic acid
  • Another object of the present invention is to provide a biomarker composition for predicting brain tumor prognosis containing, as an active ingredient, an agent capable of detecting PCNT (pericentrin) protein expression or activity level, or the expression level of a gene encoding the protein. is in
  • Another object of the present invention is to predict the prognosis of brain tumors, including the step of comparing the expression or activity level of PCNT (pericentrin) protein or the expression level of a gene encoding the protein from a sample isolated from a sample and a normal control group. It is about providing a way to present information.
  • PCNT peripheral neuronucleic acid
  • Another object of the present invention is (1) measuring the expression level of PCNT (pericentrin) protein or the expression level of a gene encoding the protein from a sample isolated from a brain tumor patient; (2) comparing the expression level of PCNT (pericentrin) protein or the expression level of the gene encoding the protein in step (1) with a control sample; And (3) a method for providing information for temozolomide and glyceofulvin combination therapy for brain tumor treatment, including determining the dose of glyceofulvin to be used in combination with temozolomide treatment is to provide
  • the present invention provides a pharmaceutical composition for treating or preventing brain tumors containing griseofulvin and temozolomide.
  • the present invention provides a biomarker composition for predicting brain tumor prognosis comprising a PCNT (pericentrin) protein or a gene encoding the protein as an active ingredient.
  • a biomarker composition for predicting brain tumor prognosis comprising a PCNT (pericentrin) protein or a gene encoding the protein as an active ingredient.
  • the present invention provides a biomarker composition for predicting brain tumor prognosis containing, as an active ingredient, an agent capable of detecting the expression or activity level of PCNT (pericentrin) protein or the expression level of a gene encoding the protein.
  • an agent capable of detecting the expression or activity level of PCNT (pericentrin) protein or the expression level of a gene encoding the protein as an active ingredient, an agent capable of detecting the expression or activity level of PCNT (pericentrin) protein or the expression level of a gene encoding the protein.
  • the present invention provides information for predicting brain tumor prognosis, including the step of comparing the expression or activity level of PCNT (pericentrin) protein or the expression level of a gene encoding the protein from a sample isolated from a sample and a normal control group. provide a way to provide PCNT (pericentrin) protein or the expression level of a gene encoding the protein from a sample isolated from a sample and a normal control group.
  • the present invention comprises the steps of (1) measuring the expression level of PCNT (pericentrin) protein or the expression level of a gene encoding the protein from a sample isolated from a brain tumor patient; (2) comparing the expression level of PCNT (pericentrin) protein or the expression level of the gene encoding the protein in step (1) with a control sample; And (3) a method for providing information for temozolomide and glyceofulvin combination therapy for brain tumor treatment, including determining the dose of glyceofulvin to be used in combination with temozolomide treatment to provide.
  • the present invention relates to a pharmaceutical composition for treating or preventing brain tumors containing griseofulvin and temozolomide. Although the effect was low for patients with temozolomide resistance, synergistic effects can be achieved by using glyceofulvin in combination.
  • Figure 2 shows the results confirmed by staining with Plk1 antibody and centrin antibody after treating human glioblastoma cell lines U87 cell line and U373 cell line with 10 ⁇ M temozolomide for 24 hours.
  • Figure 3 shows the results of confirming the cells by staining with PCNT (pericentrin) antibody and beta-tubulin antibody after treating the U87 and U373 cell lines with the drug for 24 hours.
  • Figure 4 shows the results of counting multipolar spindles and unaligned chromosomes by observing cells after antibody staining after treating 7 glioblastoma cell lines with drugs for 24 hours.
  • PCNT peripheral neurotrophin
  • the present invention provides a pharmaceutical composition for treating or preventing brain tumors containing griseofulvin and temozolomide.
  • the concentration of griseofulvin may be 0.0353 mg/kg to 3.528 mg/kg, and the concentration of temozolomide may be 0.971 mg/kg to 1.94 mg/kg, but is not limited thereto.
  • the brain tumor may be any one selected from the group consisting of glioma, low-grade glioma, glioma multiforme, glioblastoma, glioblastoma multiforme, astrocytoma, anaplastic astrocytoma, oligodendrocyte tumor, and ependymocyte tumor, It is not limited to this.
  • the pharmaceutical composition may be administered to a patient overexpressing MGMT (O(6)-methylguanine-DNA methyltransferase).
  • temozolomide weakens centrosomes to induce multipolar spindles to induce apoptosis
  • griseofulvin can induce centrosome declustering.
  • the pharmaceutical composition is a suitable carrier, excipient, disintegrant, sweetener, coating agent, swelling agent, lubricant, lubricant, flavoring agent, antioxidant, buffer, bacteriostatic agent, One or more additives selected from the group consisting of diluents, dispersants, surfactants, binders and lubricants may be further included.
  • carriers, excipients and diluents are lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, gum acacia, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methyl cellulose, microcrystalline Cellulose, polyvinyl pyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate, and mineral oil may be used, and solid dosage forms for oral administration include tablets, pills, powders, granules, and capsules.
  • solid preparations may be prepared by mixing at least one or more excipients, for example, starch, calcium carbonate, sucrose or lactose, gelatin, etc., with the composition.
  • excipients for example, starch, calcium carbonate, sucrose or lactose, gelatin, etc.
  • lubricants such as magnesium stearate and talc may also be used.
  • Liquid preparations for oral administration include suspensions, solutions for oral use, emulsions, syrups, and the like, and various excipients such as wetting agents, sweeteners, aromatics, and preservatives may be included in addition to commonly used simple diluents such as water and liquid paraffin.
  • Preparations for parenteral administration include sterilized aqueous solutions, non-aqueous solvents, suspensions, emulsions, freeze-dried preparations, suppositories, and the like.
  • Propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and injectable esters such as ethyl oleate may be used as non-aqueous solvents and suspensions.
  • As a base material of the suppository witepsol, macrogol, tween 61, cacao butter, laurin paper, glycerogeratin and the like may be used.
  • the pharmaceutical composition is intravenous, intraarterial, intraperitoneal, intramuscular, intraarterial, intraperitoneal, intrasternal, transdermal, intranasal, inhalational, topical, rectal, oral, intraocular or It can be administered to a subject in a conventional manner via the intradermal route.
  • the dosage of the active ingredient according to the present invention may vary depending on the condition and weight of the subject, the type and severity of the disease, the drug type, the route and duration of administration, and may be appropriately selected by a person skilled in the art, and the daily dosage is 0.01 mg.
  • Administration may be administered once a day or divided into several times, and the scope of the present invention is not limited thereby.
  • the present invention provides a biomarker composition for predicting brain tumor prognosis comprising a PCNT (pericentrin) protein or a gene encoding the protein as an active ingredient.
  • a biomarker composition for predicting brain tumor prognosis comprising a PCNT (pericentrin) protein or a gene encoding the protein as an active ingredient.
  • the "biomarker" of the present invention is a substance that can be diagnosed by distinguishing tissues or cells of a subject with a brain tumor from tissues or cells of a normal control group, and is a pattern of increase or decrease in tissues or cells of a subject with a disease compared to a normal control group. It includes organic biomolecules such as proteins or nucleic acids, lipids, glycolipids, glycoproteins, etc.
  • prognosis prediction refers to the act of predicting the course and outcome of a disease in advance. More specifically, prognosis prediction is interpreted as meaning that the course of a disease after treatment may vary depending on the patient's physiological or environmental state, and that it refers to any act of predicting the course of a disease after treatment by comprehensively considering the patient's condition. It can be.
  • the prediction of prognosis may be interpreted as an act of predicting the disease-free survival rate or survival rate of a cancer patient by predicting in advance the course of a disease and whether or not it is completely cured after treatment of a cancer patient.
  • predicting "good prognosis” means that the disease-free survival rate or survival rate of cancer patients after treatment is high, so that cancer patients are likely to be cured, and predicting "poor prognosis” means that cancer patients are likely to be treated. Since the disease-free survival rate or survival rate of cancer patients after treatment is at a low level, it means that there is a high possibility of cancer recurrence or death due to cancer from cancer patients.
  • the present invention provides a biomarker composition for predicting brain tumor prognosis containing, as an active ingredient, an agent capable of detecting the expression or activity level of PCNT (pericentrin) protein or the expression level of a gene encoding the protein.
  • an agent capable of detecting the expression or activity level of PCNT (pericentrin) protein or the expression level of a gene encoding the protein as an active ingredient, an agent capable of detecting the expression or activity level of PCNT (pericentrin) protein or the expression level of a gene encoding the protein.
  • the agent may be selected from the group consisting of primers, probes, antibodies, peptides and aptamers.
  • primer is a short gene sequence that is the starting point of DNA synthesis, and refers to an oligonucleotide synthesized for the purpose of diagnosis, DNA sequencing, and the like.
  • the primers may be synthesized and used in a length of 15 to 30 base pairs, but may vary depending on the purpose of use, and may be modified by methylation, capping, or the like by a known method.
  • probe refers to a nucleic acid capable of specifically binding to mRNA of several to several hundred bases in length prepared through enzymatic chemical separation and purification or synthesis. The presence or absence of mRNA can be confirmed by labeling a radioactive isotope or an enzyme, and it can be designed and modified using a known method.
  • the term “antibody” is a term known in the art and refers to a specific immunoglobulin directed against an antigenic site.
  • the antibody in the present invention refers to an antibody that specifically binds to the PCNT of the present invention, and the antibody can be prepared according to a conventional method in the art.
  • the type of antibody includes polyclonal antibodies or monoclonal antibodies, and all immunoglobulin antibodies are included.
  • the antibody is meant in its complete form with two full-length light chains and two full-length heavy chains.
  • the antibody also includes special antibodies such as humanized antibodies.
  • peptide used in the present invention has the advantage of high binding ability to a target substance, and does not undergo denaturation even during heat/chemical treatment.
  • peptide because of its small molecular size, it can be attached to other proteins and used as a fusion protein. Specifically, since it can be used by attaching it to a polymer protein chain, it can be used as a diagnostic kit and a drug delivery material.
  • aptamer is a special kind of single-stranded nucleic acid (DNA, RNA or modified nucleic acid) that has a stable tertiary structure and can bind to a target molecule with high affinity and specificity. It means a kind of composed polynucleotide. As described above, aptamers can specifically bind to antigenic substances in the same way as antibodies, but are more stable than proteins, have a simple structure, and are composed of polynucleotides that are easy to synthesize, so they can be used instead of antibodies. can
  • the present invention provides information for predicting brain tumor prognosis, including the step of comparing the expression or activity level of PCNT (pericentrin) protein or the expression level of a gene encoding the protein from a sample isolated from a sample and a normal control group. provide a way to provide PCNT (pericentrin) protein or the expression level of a gene encoding the protein from a sample isolated from a sample and a normal control group.
  • the present invention comprises the steps of (1) measuring the expression level of PCNT (pericentrin) protein or the expression level of a gene encoding the protein from a sample isolated from a brain tumor patient; (2) comparing the expression level of PCNT (pericentrin) protein or the expression level of the gene encoding the protein in step (1) with a control sample; And (3) a method for providing information for temozolomide and glyceofulvin combination therapy for brain tumor treatment, including determining the dose of glyceofulvin to be used in combination with temozolomide treatment to provide.
  • the dose of griseofulvin to be used in combination can be determined to be 0.0176 mg/kg to 0.352 mg/kg.
  • the dose of griseofulvin to be used in combination can be determined to be 0.0176 mg/kg to 0.352 mg/kg.
  • overexpression refers to a case in which the PCNT (pericentrin) protein or gene expression level is increased by 50 to 200% compared to the control group
  • low expression refers to a case in which the PCNT (pericentrin) protein or gene expression level is increased by 20 to 200% compared to the control group. This means a 50% reduction.
  • Example 1 temozolomide according to processing multipolarity Confirmation of spindle formation and mechanism of apoptosis
  • Example 3 glyceofulvin and temozolomide by combined treatment multipolarity Check for spindle formation
  • the ratio was calculated by counting the number of multipolar cells out of 100 metaphase cells, and the same experiment was repeated three times to count a total of 300 cells.
  • Glioblastoma cell lines were treated with LAP200 lysis buffer (50 mM Hepes, pH 7.4, 200 mM KCl, 0.3% NP-40, 10% glycerol, 1 mM EGTA, 1 mM MgCl 2 , 0.5 mM DTT, 0.5 ⁇ M microcystin, 10 ⁇ g/ml each of leupeptin, pepstatin). and chymostatin). Proteins were separated from each cell lysate using an SDS-PAGE gel, and the proteins in the gel were transferred to a membrane and then detected with PCNT, MAD2, MGMT, and Actin antibodies to compare the protein expression levels.
  • LAP200 lysis buffer 50 mM Hepes, pH 7.4, 200 mM KCl, 0.3% NP-40, 10% glycerol, 1 mM EGTA, 1 mM MgCl 2 , 0.5 mM DTT, 0.5 ⁇ M microcystin, 10
  • PCNT peripheral neurotrophin
  • Example 5 glyceofulvin and temozolomide by combined treatment colony Confirm colony formation

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Abstract

La présente invention concerne une composition pharmaceutique comprenant de la griséofulvine et du témozolomide pour le traitement ou la prévention de tumeurs cérébrales. Lorsque le témozolomide est utilisé seul, la prolongation de la période de survie est insignifiante et les patients résistants au témozolomide ne reçoivent que peu d'effet, mais l'utilisation de la griséofulvine en association avec celui-ci présente des effets synergiques de sorte à conduire à de meilleurs effets, et cette association peut ainsi être utilisée dans le traitement de tumeurs cérébrales.
PCT/KR2022/020588 2021-12-17 2022-12-16 Polythérapie à base de griséofulvine pour le traitement d'une tumeur cérébrale WO2023113538A1 (fr)

Applications Claiming Priority (4)

Application Number Priority Date Filing Date Title
KR20210181387 2021-12-17
KR10-2021-0181387 2021-12-17
KR10-2022-0175813 2022-12-15
KR1020220175813A KR20230092793A (ko) 2021-12-17 2022-12-15 뇌종양 치료를 위한 글리세오플빈 병용요법

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Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20040115697A1 (en) * 2002-08-09 2004-06-17 Doxsey Stephen J. Cancer diagnostics and prognostics
WO2006052976A2 (fr) * 2004-11-09 2006-05-18 Schering Corporation Methodes de traitement
US20140235556A1 (en) * 2011-09-27 2014-08-21 Biomed Valley Discoveries, Inc. Compositions and methods of treating gliomas
US20190133985A1 (en) * 2016-03-24 2019-05-09 Orbus Therapeutics, Inc. Compositions and methods for use of eflornithine and derivatives and analogs thereof to treat cancers, including gliomas
KR20200019927A (ko) * 2017-01-11 2020-02-25 의료법인 성광의료재단 뇌신경계 질환의 치료 또는 진단을 위한 표적 단백질로서 pcnt

Patent Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20040115697A1 (en) * 2002-08-09 2004-06-17 Doxsey Stephen J. Cancer diagnostics and prognostics
WO2006052976A2 (fr) * 2004-11-09 2006-05-18 Schering Corporation Methodes de traitement
US20140235556A1 (en) * 2011-09-27 2014-08-21 Biomed Valley Discoveries, Inc. Compositions and methods of treating gliomas
US20190133985A1 (en) * 2016-03-24 2019-05-09 Orbus Therapeutics, Inc. Compositions and methods for use of eflornithine and derivatives and analogs thereof to treat cancers, including gliomas
KR20200019927A (ko) * 2017-01-11 2020-02-25 의료법인 성광의료재단 뇌신경계 질환의 치료 또는 진단을 위한 표적 단백질로서 pcnt

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