[go: up one dir, main page]

WO2018190680A1 - NUTLINE-3α PERMETTANT DE MAÎTRISER MYCOBACTERIUM TUBERCULOSIS INTRACELLULAIRE, ET COMPOSITION ET MÉTHODE PERMETTANT DE RÉGULER L'EXPRESSION DE P53 - Google Patents

NUTLINE-3α PERMETTANT DE MAÎTRISER MYCOBACTERIUM TUBERCULOSIS INTRACELLULAIRE, ET COMPOSITION ET MÉTHODE PERMETTANT DE RÉGULER L'EXPRESSION DE P53 Download PDF

Info

Publication number
WO2018190680A1
WO2018190680A1 PCT/KR2018/004350 KR2018004350W WO2018190680A1 WO 2018190680 A1 WO2018190680 A1 WO 2018190680A1 KR 2018004350 W KR2018004350 W KR 2018004350W WO 2018190680 A1 WO2018190680 A1 WO 2018190680A1
Authority
WO
WIPO (PCT)
Prior art keywords
tuberculosis
nutlin
mycobacterium tuberculosis
mycobacterium
pharmaceutical composition
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Ceased
Application number
PCT/KR2018/004350
Other languages
English (en)
Korean (ko)
Inventor
송창화
임윤지
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Industry and Academy Cooperation In Chungnam National University
Original Assignee
Industry and Academy Cooperation In Chungnam National University
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Priority claimed from KR1020180041520A external-priority patent/KR102082285B1/ko
Application filed by Industry and Academy Cooperation In Chungnam National University filed Critical Industry and Academy Cooperation In Chungnam National University
Publication of WO2018190680A1 publication Critical patent/WO2018190680A1/fr
Anticipated expiration legal-status Critical
Ceased legal-status Critical Current

Links

Images

Classifications

    • 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
    • A61K31/13—Amines
    • 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
    • A61K31/435—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom
    • A61K31/44—Non condensed pyridines; Hydrogenated derivatives thereof
    • A61K31/4409—Non condensed pyridines; Hydrogenated derivatives thereof only substituted in position 4, e.g. isoniazid, iproniazid
    • 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
    • A61K31/496—Non-condensed piperazines containing further heterocyclic rings, e.g. rifampin, thiothixene or sparfloxacin

Definitions

  • the present invention relates generally to the field of immunology and, in part, to methods of inhibiting survival and proliferation of Mycobacterium tuberculosis and to methods for providing Nutlin-3 ⁇ (neutrin-3 ⁇ ) and p53 expression control compositions.
  • Tuberculosis is a chronic infectious disease caused by infection of Mycobacterium tuberculosis and other Mycobacterium species. Tuberculosis is a major disease in developing countries that kills about 8 million new infections each year, killing 3 million lives, and is also a problem in developed countries. While the infection may be asymptomatic for a significant period of time, tuberculosis is the most common symptom, indicating acute inflammation of the lung, resulting in fever and nonproductive cough. If left untreated, it usually results in serious complications and death.
  • Tuberculosis can be treated using prolonged antibiotic therapy, but this treatment is not sufficient to prevent the spread of tuberculosis. Infected individuals do not exhibit symptoms but may be contagious for a period of time. In addition, it is difficult to control the patient's behavior even if the treatment regimen is strictly followed. Some patients do not complete the course of treatment, which can lead to ineffective treatment and development of drug resistance. Even when the entire course of treatment is completed, the TB germ infection is not eradicated from the infected individual and remains a latent infection that can still be reactivated.
  • BCG Bacillus Calmette-Guerin
  • PPD tuberculin proteinpurified derivative
  • Antigen-specific T cell responses result in measurable degree induration at the site of infusion up to 48-72 hours post infusion, indicating exposure to mycobacterium antigens.
  • the assay is problematic in sensitivity and specificity, and individuals vaccinated with BCG cannot be distinguished from those infected.
  • p53 molecular weight 53 kDa
  • PUMA molecular weight 53 kDa
  • the P53 protein is labeled ubiquitin by MDM2 protein and degraded by proteasome.
  • Nutlin-3 ⁇ (molecular weight 581.49), known as an anticancer agent, binds to this MDM2 and inhibits p53-MDM2 interaction to increase p53 pathway activity. to be. Therefore, as a method for inhibiting Mycobacterium tuberculosis with p53 as a target, Nutlin-3 ⁇ alone or in combination with existing anti-tuberculosis therapeutics may be used.
  • the present invention contributes to the activation of macrophages using Nutlin-3 ⁇ , which induces expression of p53 or induces p53 activity, to provide a method for preventing and effectively treating tuberculosis, which has a different mechanism of action and relatively fewer side effects.
  • the present invention is to provide a pharmaceutical composition for preventing and treating tuberculosis comprising a Nutlin-3 ⁇ compound as an active ingredient.
  • the Nutlin-3 ⁇ compound thus provided is characterized by overexpressing p53, and the overexpressed p53 inhibits the growth of Mycobacterium tuberculosis by inducing apoptosis of macrophages infected with Mycobacterium tuberculosis.
  • the anti-tuberculosis agent used herein may be at least one selected from the group consisting of Rifampicin, Isoniazid, Ethambutol, and Pyrazinamide.
  • the present invention utilizes the function of Mycobacterium tuberculosis of Nutlin-3 ⁇ , which is not known at all, it can be expected to have a prophylactic and therapeutic effect against Mycobacterium tuberculosis having multidrug resistance with respect to existing drugs.
  • the growth of latent Mycobacterium tuberculosis can be suppressed, so that latent Mycobacterium tuberculosis can be removed when used alone or in combination with a conventional medicine.
  • Figure 2 is a protein expression electrophoresis picture by western blotting showing p53 expression and Caspase-3 activity according to Nutlin-3 ⁇ concentrations treated in Mycobacterium tuberculosis-infected macrophages.
  • 3 is a graph showing that the number of Mycobacterium tuberculosis in macrophages is significantly reduced by Nutlin-3 ⁇ treatment.
  • 4 and 5 are graphs showing that the number of Mycobacterium tuberculosis in macrophages is significantly reduced by the combination of antituberculosis therapeutic agents (isoniazid, rifampicin, pyrazinamide, ethambutol) and Nutlin-3 ⁇ .
  • antituberculosis therapeutic agents isoniazid, rifampicin, pyrazinamide, ethambutol
  • Nutlin-3 ⁇ are graphs showing that the number of Mycobacterium tuberculosis in macrophages is significantly reduced by the combination of antituberculosis therapeutic agents (isoniazid, rifampicin, pyrazinamide, ethambutol) and Nutlin-3 ⁇ .
  • FIG. 6 is a graph showing a significant increase in Mycobacterium tuberculosis survival in mice lungs with p53-deficient macrophage.
  • FIG. 7 is a graph showing that the number of Mycobacterium tuberculosis in lung tissue is significantly decreased when pneumoniae-infected mice are intraperitoneally injected with Nutlin-3a (10 mg / kg / day).
  • FIG. 8 is a graph showing that the expression of p53 in the macrophage of tuberculosis patients is significantly lower than that of healthy macrophage.
  • FIG. 9 is a graph showing that the number of Mycobacterium tuberculosis in cells is significantly reduced when p53 activity is increased through Nutlin-3a.
  • the present invention was first identified that the Nutlin-3 ⁇ compound has an effect of preventing or treating Mycobacterium tuberculosis through p53 overexpression and acting on apoptosis of macrophages.
  • the present invention comprises a Nutlin-3 ⁇ compound as an active ingredient. It is characterized by providing a composition for preventing or treating Mycobacterium tuberculosis.
  • 'Nutlin-3 ⁇ ' is a small molecule compound that binds to MDM2 and inhibits binding between MDM2 and p53 protein, and is known for its anticancer effect, and its chemical structure is shown in FIG. 1. More specifically, Nutlin-3 ⁇ is a small molecule compound that binds to MDM2 and inhibits binding between MDM2 and p53 protein. It is an anticancer drug currently being developed for the treatment of solid cancer. In detail, the MDM2 protein binds to the transactivation site of p53, a gene that inhibits cancer, and blocks the activation ability. The Nutlin-3 ⁇ compound has a molecular structure similar to that of p53.
  • Nutlin-3 ⁇ compound is only known as an anticancer drug that is currently being developed in the clinic, and there is no report on the effect that it has on the treatment of tuberculosis.
  • the present inventors have completed the present invention by confirming that Nutlin-3 ⁇ compound can improve tuberculosis through a mechanism of activating p53.
  • Example 1 of the present invention when the Nutlin-3 ⁇ compound was injected into Bone marrow-derived macrophages (BMDMs) cells, which are mouse-derived macrophages, the number of Mycobacterium tuberculosis bacteria was significantly reduced, resulting in an improvement in tuberculosis. It could be confirmed.
  • BMDMs Bone marrow-derived macrophages
  • Example 2 of the present invention when used in combination with the existing anti-tuberculosis agent was confirmed to show an effect of significantly higher tuberculosis bacteria compared to the use of the existing anti-tuberculosis agent alone.
  • Nutlin-3 ⁇ compound is effective in improving tuberculosis
  • the composition of the present invention comprising it as an active ingredient can be usefully used in the prevention or treatment for tuberculosis.
  • Tuberculosis in the present invention includes eye tuberculosis, skin tuberculosis, adrenal tuberculosis, kidney tuberculosis, epididymal tuberculosis, lymphatic tuberculosis, laryngeal tuberculosis, middle ear tuberculosis, intestinal tuberculosis, multidrug-resistant tuberculosis, pulmonary tuberculosis, tuberculosis, bone tuberculosis, throat tuberculosis, lymphatic tuberculosis We can include ruins, breast tuberculosis or spinal tuberculosis.
  • existing anti-tuberculosis agents may include Rifampicin, Isoniazid, Ethambutol or Pyrazinamide used in Example 2 of the present invention.
  • the term 'treatment' unless stated otherwise, reverses, alleviates, inhibits, or prevents the disease or condition to which the term applies, or one or more symptoms of the disease or condition, As used herein, the term 'treatment' refers to the act of treating when 'treating' is defined as above.
  • BMDMs Bone marrow-derived macrophages
  • Mycobacterium tuberculosis H37Ra non-pathogenic Mycobacterium tuberculosis
  • Figure 2 shows the expression level of p53 protein expressed during Mycobacterium tuberculosis infection and the apoptosis marker Caspase-3 activity as a result of Western blot method in accordance with the increased treatment concentration of Nutlin-3 ⁇ (Fig. 1), a p53 inducing agent in macrophages infected with Mycobacterium tuberculosis. Increasing p53 and Caspas-3 activity was observed. ( ⁇ -actin is an internal control in Figure 2)
  • FIG. 3 was treated with Nutlin-3 ⁇ (30 uM) in large macrophages infected with Mycobacterium tuberculosis and after 24 hours, the number of viable Mycobacterium tuberculosis cells was cultured in 7H10 agar medium for 14-21 days. As a result, the number of survival of the intracellular pathogenic Mycobacterium tuberculosis and nonpathogenic Mycobacterium tuberculosis was significantly reduced in the macrophages treated with Nutlin-3 ⁇ (30 uM) compared with the control group.
  • Figures 4 and 5 show anti-tuberculosis treatment of isoniazid (10 ng / ml), rifampicin (10 ng / ml), pyrazinamide (100 ng / ml) or etambutol (100 ng / ml) during the infection of Mycobacterium tuberculosis bacteria. And Nutlin-3 ⁇ (30 uM) in combination to inhibit the growth of mycobacterium tuberculosis.
  • Macrophage cells infected with pathogenic Mycobacterium tuberculosis or non-pathogenic Mycobacterium tuberculosis were treated with antituberculosis treatment alone or in combination with Nutlin-3 ⁇ to determine the number of tuberculosis viable in cells after 24 hours.
  • the experimental results showed that the number of intracellular tuberculosis bacteria decreased significantly when anti-TB treatment and Nutlin-3 ⁇ were used in combination with anti-TB treatment alone.
  • FIG. 6 shows the number of viable Mycobacterium tuberculosis bacteria in lung tissue after intranasal infection of non-pathogenic Mycobacterium tuberculosis in a control (wild type) or macrophage-deficient p53 (p53 conditional knockout) mouse.
  • wild type wild type
  • macrophage-deficient p53 p53 conditional knockout
  • FIG. 7 shows that the number of Mycobacterium tuberculosis in lung tissue was significantly reduced when p53 activity was increased by intraperitoneal injection of Nutlin-3a (10 mg / kg / day) into mice infected with non-pathogenic Mycobacterium tuberculosis via the nasal cavity.
  • the expression of p53 in the macrophage of tuberculosis patients was significantly lower than that of healthy macrophage.
  • Figure 9 was infected with non-pathogenic Mycobacterium tuberculosis in macrophage derived from the blood of healthy or tuberculosis patients, treated with Nutlin-3a (10 uM) and the number of intracellular Mycobacterium viability was measured. As a result, when the p53 activity was increased through Nutlin-3a, it was confirmed that the number of Mycobacterium tuberculosis in the cells was significantly reduced.
  • the mycobacterium tuberculosis can be suppressed by increasing the expression of p53 in the cells. Therefore, when the composition according to the present invention (Nutlin-3 ⁇ ) is treated with Mycobacterium tuberculosis-infected cells as an active ingredient to control p53, that is, when the composition of the present invention is treated to Mycobacterium tuberculosis carriers, tuberculosis patients or patients with existing anti-tuberculosis drug resistance It can be expected to kill my tuberculosis bacteria.
  • the statistical method used was Bonferroni's multiple comparison test, marked with * when p-value ⁇ 0.05, ** when p-value ⁇ 0.01, and *** when p-value ⁇ 0.001.

Landscapes

  • Health & Medical Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Medicinal Chemistry (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Epidemiology (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Animal Behavior & Ethology (AREA)
  • General Health & Medical Sciences (AREA)
  • Public Health (AREA)
  • Veterinary Medicine (AREA)
  • Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
  • Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)

Abstract

La présente invention concerne une composition pharmaceutique permettant de prévenir ou de traiter la tuberculose, comprenant comme principe actif un composé à base de Nutline-3α. Un composé à base de Nutline-3α, qui est un principe actif selon la présente invention, a pour effet d'inhiber la prolifération de Mycobacterium tuberculosis en induisant, par l'intermédiaire de la surexpression de p53, l'apoptose des macrophages qui ont été infectés par Mycobacterium tuberculosis. Ainsi, une méthode selon la présente invention comprenant le composé à base de Nutline-3α comme principe actif peut être utilisée de manière utile pour prévenir ou traiter la tuberculose.
PCT/KR2018/004350 2017-04-13 2018-04-13 NUTLINE-3α PERMETTANT DE MAÎTRISER MYCOBACTERIUM TUBERCULOSIS INTRACELLULAIRE, ET COMPOSITION ET MÉTHODE PERMETTANT DE RÉGULER L'EXPRESSION DE P53 Ceased WO2018190680A1 (fr)

Applications Claiming Priority (4)

Application Number Priority Date Filing Date Title
KR10-2017-0047868 2017-04-13
KR20170047868 2017-04-13
KR10-2018-0041520 2018-04-10
KR1020180041520A KR102082285B1 (ko) 2017-04-13 2018-04-10 세포내 결핵균 제어를 위한 Nutlin-3α (뉴트린-3α) 및 p53 발현 조절 조성물 또는 방법

Publications (1)

Publication Number Publication Date
WO2018190680A1 true WO2018190680A1 (fr) 2018-10-18

Family

ID=63793436

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/KR2018/004350 Ceased WO2018190680A1 (fr) 2017-04-13 2018-04-13 NUTLINE-3α PERMETTANT DE MAÎTRISER MYCOBACTERIUM TUBERCULOSIS INTRACELLULAIRE, ET COMPOSITION ET MÉTHODE PERMETTANT DE RÉGULER L'EXPRESSION DE P53

Country Status (1)

Country Link
WO (1) WO2018190680A1 (fr)

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN112575078A (zh) * 2020-12-30 2021-03-30 山东大学第二医院 lncRNAs作为活动性肺结核病特异性标志物的应用
US20210379057A1 (en) * 2018-10-16 2021-12-09 Massachusetts Institute Of Technology Nutlin-3a for use in treating a mycobacterium tuberculosis infection

Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
KR20110063518A (ko) * 2008-09-03 2011-06-10 화이자 인코포레이티드 결핵에 대한 병용 요법
KR20140032312A (ko) * 2012-09-06 2014-03-14 충남대학교산학협력단 결핵 치료용 약학적 조성물
KR20150128731A (ko) * 2013-03-15 2015-11-18 보드 오브 리전츠, 더 유니버시티 오브 텍사스 시스템 누트린-3a 및 펩티드에 의한 폐 섬유증의 저해

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
KR20110063518A (ko) * 2008-09-03 2011-06-10 화이자 인코포레이티드 결핵에 대한 병용 요법
KR20140032312A (ko) * 2012-09-06 2014-03-14 충남대학교산학협력단 결핵 치료용 약학적 조성물
KR20150128731A (ko) * 2013-03-15 2015-11-18 보드 오브 리전츠, 더 유니버시티 오브 텍사스 시스템 누트린-3a 및 펩티드에 의한 폐 섬유증의 저해

Non-Patent Citations (2)

* Cited by examiner, † Cited by third party
Title
CRANE, E. K.: "Nutlin-3a: A potential therapeutic opportunity for TP53 wild-type ovarian carcinomas", PLOS ONE, 2015, pages 1 - 13 *
MADENSPACHER, J. H.: "p53 integrates host defense and cell fate during bacterial pneumonia", THE JOURNAL OF EXPERIMENTAL MEDICINE, 2013, pages 891 - 904 *

Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20210379057A1 (en) * 2018-10-16 2021-12-09 Massachusetts Institute Of Technology Nutlin-3a for use in treating a mycobacterium tuberculosis infection
CN112575078A (zh) * 2020-12-30 2021-03-30 山东大学第二医院 lncRNAs作为活动性肺结核病特异性标志物的应用
CN112575078B (zh) * 2020-12-30 2022-10-04 山东大学第二医院 lncRNAs作为活动性肺结核病特异性标志物的应用

Similar Documents

Publication Publication Date Title
Martins et al. The curative activity of thioridazine on mice infected with Mycobacterium tuberculosis
Yang et al. A tobramycin vector enhances synergy and efficacy of efflux pump inhibitors against multidrug-resistant Gram-negative bacteria
Elkashif et al. Investigation of auranofin and gold-containing analogues antibacterial activity against multidrug-resistant Neisseria gonorrhoeae
Thomas et al. Galleria mellonella as a model system to test the pharmacokinetics and efficacy of antibiotics against Burkholderia pseudomallei
Legendre et al. Hansen's disease (leprosy): current and future pharmacotherapy and treatment of disease‐related immunologic reactions
Zhou et al. Trained immunity contributes to the prevention of Mycobacterium tuberculosis infection, a novel role of autophagy
Wang et al. Baicalein resensitizes multidrug-resistant gram-negative pathogens to doxycycline
J. Barton et al. Host restriction, pathogenesis and chronic carriage of typhoidal Salmonella
Holloway et al. Cystatins 9 and C as a novel immunotherapy treatment that protects against multidrug-resistant New Delhi metallo-beta-lactamase-1-producing Klebsiella pneumoniae
Lu et al. Antibacterial activity of an FtsZ inhibitor celastrol and its synergistic effect with vancomycin against enterococci in vitro and in vivo
Wang et al. FDA-approved amoxapine effectively promotes macrophage control of mycobacteria by inducing autophagy
WO2018190680A1 (fr) NUTLINE-3α PERMETTANT DE MAÎTRISER MYCOBACTERIUM TUBERCULOSIS INTRACELLULAIRE, ET COMPOSITION ET MÉTHODE PERMETTANT DE RÉGULER L'EXPRESSION DE P53
Park et al. Understanding the reciprocal interplay between antibiotics and host immune system: how can we improve the anti-mycobacterial activity of current drugs to better control tuberculosis?
Wu et al. Effects of l-Serine on Macrolide Resistance in Streptococcus suis
WO2018008932A1 (fr) Composition visant à inhiber la survie ou la croissance du bacille de la tuberculose par réglage de la polarisation des macrophages, et méthode utilisant cette dernière
KR102082285B1 (ko) 세포내 결핵균 제어를 위한 Nutlin-3α (뉴트린-3α) 및 p53 발현 조절 조성물 또는 방법
KR102039696B1 (ko) 세포내 결핵균 제어를 위한 p53 발현 조절 가능한 조성물 (K279-1558) 또는 방법
Chen et al. Inhibitory and injury-protection effects of O-glycan on gastric epithelial cells infected with Helicobacter pylori
Mir et al. Immunotherapeutic potential of N-formylated peptides of ESAT-6 and glutamine synthetase in experimental tuberculosis
Suzuki et al. Buruli ulcer in Japan
KR20180031277A (ko) Scotin 단백질의 과발현에 의하여 결핵균 생존 및 증식을 억제하는 방법
Guo et al. Honokiol inhibits Vibrio harveyi hemolysin virulence by reducing its haemolytic activity
KR101935292B1 (ko) 대식세포의 미토콘드리아 막 단백질 mfn2의 녹다운을 이용한 결핵균 억제방법
Naamara et al. Treating chancroid with enoxacin.
Singh et al. Tuberculosis: introduction, drug regimens, and multidrug-resistance

Legal Events

Date Code Title Description
121 Ep: the epo has been informed by wipo that ep was designated in this application

Ref document number: 18784167

Country of ref document: EP

Kind code of ref document: A1

NENP Non-entry into the national phase

Ref country code: DE

122 Ep: pct application non-entry in european phase

Ref document number: 18784167

Country of ref document: EP

Kind code of ref document: A1