CN111166750B - 一种4-氟-2-甲基吲哚类化合物的抗菌新用途 - Google Patents
一种4-氟-2-甲基吲哚类化合物的抗菌新用途 Download PDFInfo
- Publication number
- CN111166750B CN111166750B CN201811330994.8A CN201811330994A CN111166750B CN 111166750 B CN111166750 B CN 111166750B CN 201811330994 A CN201811330994 A CN 201811330994A CN 111166750 B CN111166750 B CN 111166750B
- Authority
- CN
- China
- Prior art keywords
- fluoro
- compound
- methylindole
- mycobacterium tuberculosis
- hydroxy
- 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.)
- Active
Links
- -1 4-fluoro-2-methylindole compound Chemical class 0.000 title claims abstract description 10
- 230000000844 anti-bacterial effect Effects 0.000 title abstract description 9
- 241000187479 Mycobacterium tuberculosis Species 0.000 claims abstract description 18
- 241000191963 Staphylococcus epidermidis Species 0.000 claims abstract description 15
- 150000001875 compounds Chemical class 0.000 claims abstract description 15
- 241000187480 Mycobacterium smegmatis Species 0.000 claims abstract description 10
- XXJWYDDUDKYVKI-UHFFFAOYSA-N 4-[(4-fluoro-2-methyl-1H-indol-5-yl)oxy]-6-methoxy-7-[3-(1-pyrrolidinyl)propoxy]quinazoline Chemical compound COC1=CC2=C(OC=3C(=C4C=C(C)NC4=CC=3)F)N=CN=C2C=C1OCCCN1CCCC1 XXJWYDDUDKYVKI-UHFFFAOYSA-N 0.000 claims description 7
- 229960002412 cediranib Drugs 0.000 claims description 7
- 229940124350 antibacterial drug Drugs 0.000 claims description 5
- WCWUXEGQKLTGDX-LLVKDONJSA-N (2R)-1-[[4-[(4-fluoro-2-methyl-1H-indol-5-yl)oxy]-5-methyl-6-pyrrolo[2,1-f][1,2,4]triazinyl]oxy]-2-propanol Chemical compound C1=C2NC(C)=CC2=C(F)C(OC2=NC=NN3C=C(C(=C32)C)OC[C@H](O)C)=C1 WCWUXEGQKLTGDX-LLVKDONJSA-N 0.000 claims description 3
- 241000192125 Firmicutes Species 0.000 claims 2
- PHGHGDMLJZDQIR-UHFFFAOYSA-N 4-fluoro-2-methyl-1h-indole Chemical compound C1=CC=C2NC(C)=CC2=C1F PHGHGDMLJZDQIR-UHFFFAOYSA-N 0.000 claims 1
- 239000003814 drug Substances 0.000 abstract description 25
- 229940079593 drug Drugs 0.000 abstract description 20
- 230000000694 effects Effects 0.000 abstract description 17
- 230000005764 inhibitory process Effects 0.000 abstract description 9
- 102000004317 Lyases Human genes 0.000 abstract description 3
- 108090000856 Lyases Proteins 0.000 abstract description 3
- 239000005018 casein Substances 0.000 abstract description 3
- 235000021240 caseins Nutrition 0.000 abstract description 3
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 description 21
- 238000012360 testing method Methods 0.000 description 12
- UMWRMOYYUHIPDT-UHFFFAOYSA-N 4-fluoro-2-methyl-1h-indol-5-ol Chemical class OC1=CC=C2NC(C)=CC2=C1F UMWRMOYYUHIPDT-UHFFFAOYSA-N 0.000 description 11
- 239000000243 solution Substances 0.000 description 11
- 102000004190 Enzymes Human genes 0.000 description 8
- 108090000790 Enzymes Proteins 0.000 description 8
- 230000002401 inhibitory effect Effects 0.000 description 7
- 229960003907 linezolid Drugs 0.000 description 7
- TYZROVQLWOKYKF-ZDUSSCGKSA-N linezolid Chemical compound O=C1O[C@@H](CNC(=O)C)CN1C(C=C1F)=CC=C1N1CCOCC1 TYZROVQLWOKYKF-ZDUSSCGKSA-N 0.000 description 7
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 6
- 230000001580 bacterial effect Effects 0.000 description 6
- 238000000338 in vitro Methods 0.000 description 6
- 241000894006 Bacteria Species 0.000 description 5
- 230000009471 action Effects 0.000 description 5
- 239000001963 growth medium Substances 0.000 description 5
- 238000002360 preparation method Methods 0.000 description 5
- 206010011409 Cross infection Diseases 0.000 description 4
- 206010059866 Drug resistance Diseases 0.000 description 4
- 239000003112 inhibitor Substances 0.000 description 4
- 238000009630 liquid culture Methods 0.000 description 4
- 201000008827 tuberculosis Diseases 0.000 description 4
- 230000003115 biocidal effect Effects 0.000 description 3
- 238000006243 chemical reaction Methods 0.000 description 3
- 239000003153 chemical reaction reagent Substances 0.000 description 3
- 238000012258 culturing Methods 0.000 description 3
- 239000004744 fabric Substances 0.000 description 3
- 230000036541 health Effects 0.000 description 3
- 239000007788 liquid Substances 0.000 description 3
- 230000007246 mechanism Effects 0.000 description 3
- 239000002609 medium Substances 0.000 description 3
- 201000009671 multidrug-resistant tuberculosis Diseases 0.000 description 3
- 244000052769 pathogen Species 0.000 description 3
- 239000013641 positive control Substances 0.000 description 3
- 235000018102 proteins Nutrition 0.000 description 3
- 102000004169 proteins and genes Human genes 0.000 description 3
- 108090000623 proteins and genes Proteins 0.000 description 3
- JQXXHWHPUNPDRT-WLSIYKJHSA-N rifampicin Chemical compound O([C@](C1=O)(C)O/C=C/[C@@H]([C@H]([C@@H](OC(C)=O)[C@H](C)[C@H](O)[C@H](C)[C@@H](O)[C@@H](C)\C=C\C=C(C)/C(=O)NC=2C(O)=C3C([O-])=C4C)C)OC)C4=C1C3=C(O)C=2\C=N\N1CC[NH+](C)CC1 JQXXHWHPUNPDRT-WLSIYKJHSA-N 0.000 description 3
- 229960001225 rifampicin Drugs 0.000 description 3
- 229920001817 Agar Polymers 0.000 description 2
- CURLTUGMZLYLDI-UHFFFAOYSA-N Carbon dioxide Chemical compound O=C=O CURLTUGMZLYLDI-UHFFFAOYSA-N 0.000 description 2
- 206010029803 Nosocomial infection Diseases 0.000 description 2
- PLXBWHJQWKZRKG-UHFFFAOYSA-N Resazurin Chemical compound C1=CC(=O)C=C2OC3=CC(O)=CC=C3[N+]([O-])=C21 PLXBWHJQWKZRKG-UHFFFAOYSA-N 0.000 description 2
- 241000191940 Staphylococcus Species 0.000 description 2
- 239000008272 agar Substances 0.000 description 2
- GXJABQQUPOEUTA-RDJZCZTQSA-N bortezomib Chemical compound C([C@@H](C(=O)N[C@@H](CC(C)C)B(O)O)NC(=O)C=1N=CC=NC=1)C1=CC=CC=C1 GXJABQQUPOEUTA-RDJZCZTQSA-N 0.000 description 2
- 229960001467 bortezomib Drugs 0.000 description 2
- 101150043719 clpP1 gene Proteins 0.000 description 2
- 238000011161 development Methods 0.000 description 2
- 238000007865 diluting Methods 0.000 description 2
- 239000012895 dilution Substances 0.000 description 2
- 238000010790 dilution Methods 0.000 description 2
- 238000011534 incubation Methods 0.000 description 2
- 238000005259 measurement Methods 0.000 description 2
- 238000000034 method Methods 0.000 description 2
- 239000012452 mother liquor Substances 0.000 description 2
- 238000012216 screening Methods 0.000 description 2
- 238000007789 sealing Methods 0.000 description 2
- 150000003384 small molecules Chemical class 0.000 description 2
- 230000001954 sterilising effect Effects 0.000 description 2
- 239000000758 substrate Substances 0.000 description 2
- 101710197207 2-iminoacetate synthase Proteins 0.000 description 1
- 241001631457 Cannula Species 0.000 description 1
- 208000035473 Communicable disease Diseases 0.000 description 1
- 101100113734 Cyanophora paradoxa clpP-A gene Proteins 0.000 description 1
- RJQXTJLFIWVMTO-TYNCELHUSA-N Methicillin Chemical compound COC1=CC=CC(OC)=C1C(=O)N[C@@H]1C(=O)N2[C@@H](C(O)=O)C(C)(C)S[C@@H]21 RJQXTJLFIWVMTO-TYNCELHUSA-N 0.000 description 1
- 241001646725 Mycobacterium tuberculosis H37Rv Species 0.000 description 1
- 108700035964 Mycobacterium tuberculosis HsaD Proteins 0.000 description 1
- 108091005804 Peptidases Proteins 0.000 description 1
- 239000004365 Protease Substances 0.000 description 1
- 102100037486 Reverse transcriptase/ribonuclease H Human genes 0.000 description 1
- 241000191967 Staphylococcus aureus Species 0.000 description 1
- 230000003213 activating effect Effects 0.000 description 1
- 239000003242 anti bacterial agent Substances 0.000 description 1
- 238000011482 antibacterial activity assay Methods 0.000 description 1
- 229940088710 antibiotic agent Drugs 0.000 description 1
- 230000003385 bacteriostatic effect Effects 0.000 description 1
- 239000011324 bead Substances 0.000 description 1
- 229910002092 carbon dioxide Inorganic materials 0.000 description 1
- 239000001569 carbon dioxide Substances 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 230000001684 chronic effect Effects 0.000 description 1
- 210000001268 chyle Anatomy 0.000 description 1
- 238000001816 cooling Methods 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
- 208000015355 drug-resistant tuberculosis Diseases 0.000 description 1
- 230000002255 enzymatic effect Effects 0.000 description 1
- 230000005284 excitation Effects 0.000 description 1
- 208000036984 extensively drug-resistant tuberculosis Diseases 0.000 description 1
- 239000011521 glass Substances 0.000 description 1
- 230000007236 host immunity Effects 0.000 description 1
- 230000007062 hydrolysis Effects 0.000 description 1
- 238000006460 hydrolysis reaction Methods 0.000 description 1
- 208000015181 infectious disease Diseases 0.000 description 1
- 238000011081 inoculation Methods 0.000 description 1
- 231100001231 less toxic Toxicity 0.000 description 1
- 210000004072 lung Anatomy 0.000 description 1
- 229960003085 meticillin Drugs 0.000 description 1
- 244000000010 microbial pathogen Species 0.000 description 1
- 239000000178 monomer Substances 0.000 description 1
- 210000004400 mucous membrane Anatomy 0.000 description 1
- 230000008520 organization Effects 0.000 description 1
- 230000001717 pathogenic effect Effects 0.000 description 1
- 230000007918 pathogenicity Effects 0.000 description 1
- 230000002688 persistence Effects 0.000 description 1
- 239000002504 physiological saline solution Substances 0.000 description 1
- 239000000843 powder Substances 0.000 description 1
- 230000008569 process Effects 0.000 description 1
- 108090000765 processed proteins & peptides Proteins 0.000 description 1
- 230000004952 protein activity Effects 0.000 description 1
- 239000012460 protein solution Substances 0.000 description 1
- 239000011535 reaction buffer Substances 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 239000002904 solvent Substances 0.000 description 1
- 230000007480 spreading Effects 0.000 description 1
- 239000011550 stock solution Substances 0.000 description 1
- 239000006228 supernatant Substances 0.000 description 1
- 239000000725 suspension Substances 0.000 description 1
- 238000010267 two-fold dilution method Methods 0.000 description 1
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 1
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
- A61K31/33—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
-
- 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
- A61K31/41—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having five-membered rings with two or more ring hetero atoms, at least one of which being nitrogen, e.g. tetrazole
- A61K31/42—Oxazoles
- A61K31/421—1,3-Oxazoles, e.g. pemoline, trimethadione
-
- 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
- A61K31/505—Pyrimidines; Hydrogenated pyrimidines, e.g. trimethoprim
- A61K31/517—Pyrimidines; Hydrogenated pyrimidines, e.g. trimethoprim ortho- or peri-condensed with carbocyclic ring systems, e.g. quinazoline, perimidine
-
- 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
- A61K31/53—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with three nitrogens as the only ring hetero atoms, e.g. chlorazanil, melamine
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
- A61P31/02—Local antiseptics
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
- A61P31/04—Antibacterial agents
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P35/00—Antineoplastic agents
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
- Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Veterinary Medicine (AREA)
- Chemical & Material Sciences (AREA)
- Public Health (AREA)
- Medicinal Chemistry (AREA)
- General Health & Medical Sciences (AREA)
- Animal Behavior & Ethology (AREA)
- Pharmacology & Pharmacy (AREA)
- Epidemiology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Organic Chemistry (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- General Chemical & Material Sciences (AREA)
- Oncology (AREA)
- Communicable Diseases (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
本发明公开了一种4‑氟‑2‑甲基吲哚类化合物的新用途，属于生物医药领域，该类化合物能够显著抑制结核分枝杆菌酪蛋白裂解酶ClpP1P2的活性，同时能够显著抑制结核分枝杆菌以及耻垢分枝杆菌的生长，且对表皮葡萄球菌的生长也表现出一定抑制作用，因此其可用于抗菌领域药物的制备。
Description
技术领域
本发明属于生物医药技术领域，特别涉及4-氟-5-羟基-2-甲基吲哚类化合物作为酪蛋白酶裂解酶ClpP抑制剂在抗菌药物领域的应用。
背景技术
致病微生物引起的感染性疾病一直都是危害人类健康的重要因素之一。伴随着抗生素的滥用，多种抗生素耐药菌已经产生。据世界卫生组织WHO统计，全球大约有1/3的人口携带结核分枝杆菌，大约有860万例结核病患者，每年约170万人死于该病，而多药耐药结核（multidrug-resistant tuberculosis, MDR-TB）和广泛耐药结核（extensively drug-resistant tuberculosis, XDR-TB）的出现，为结核病的治疗带来更严峻的挑战。
表皮葡萄球菌是一种革兰阴性的葡萄球菌，一般黏附于人体皮肤和黏膜，是重要的条件致病菌。近年来，随着插管、人工瓣膜、人工关节等医疗移植物的广泛使用，由表皮葡萄球菌（Staphylococcus epidermidis）引起的医院感染日趋严重，其中凝固酶阴性葡萄球菌占医院感染病原中的第 1 位，目前已成为医院感染的重要病原菌之一。由于表皮葡萄球菌分泌的毒性因子较金黄葡萄球菌少，因此表皮葡萄球菌的致病性主要与其在医疗移植物表面形成的生物被膜（Biofilm）有关，它可以抵抗宿主免疫力和增强细菌耐药性，从而导致感染呈现慢性、持续性和反复性的特点，很难治愈，而表皮葡萄球菌对常见抗菌药物耐药率已成为医院感染和临床治疗的棘手问题。因此迫切需要发现全新作用机制的新型抗菌药物来应对临床治疗中的耐药问题。
酪氨酸裂解酶ClpP是一个非常重要且作用机制新颖的抗生素作用靶点，其激动和抑制形式，均能够在不同细菌中发挥其抗菌活性，是一种全新的抗生素机制，为临床治疗中耐药问题的解决带来了希望。
发明内容
本发明要解决的技术问题是发现4-氟-5-羟基-2-甲基吲哚类化合物的抗菌新用途，可用于抗菌药物的制备。为解决上述技术问题，本发明通过如下技术方式实现：本发明中4-氟-5-羟基-2-甲基吲哚类化合物在体外酶活筛选模型上，能够显著抑制结核分枝杆菌酪蛋白裂解酶ClpP1P2活性，因此可作为其抑制剂进行药物制备；此外，抗菌活性测定（MIC）结果显示4-氟-5-羟基-2-甲基吲哚类化合物对结核分枝杆菌以及耻垢分枝杆菌的生长有明显的抑制作用，同时对表皮葡萄球菌的生长也表现出一定的抑制作用，因此，4-氟-5-羟基-2-甲基吲哚类化合物可作为结核分枝杆菌ClpP1P2抑制剂用于抗菌领域的药物制备。
附图说明
图1 是本发明实施例1中不同作用时间及不同浓度的4-氟-5-羟基-2-甲基吲哚类化合物对结核分枝杆菌ClpP1P2酶活的影响。
具体实施方式
以下结合实施例对本发明做进一步阐述。
实施例1 4-氟-5-羟基-2-甲基吲哚类化合物对结核分枝杆菌ClpP1P2酶水平活性影响
本发明自行建立结核分枝杆菌ClpP1P2蛋白活性筛选模型，当有benzoyl-leucine-leucine（Bz-L-L）存在下，ClpP1P2蛋白特异水解带有荧光基团的底物Z-Gly-Gly-Leu-AMC，通过检测该荧光短肽被ClpP1P2水解过程产生的荧光信号的强弱来评价小分子对该酶活性的影响。
1）蛋白样品准备：ClpP1/ClpP2单体以摩尔比1:1混合，加入终浓度为2.5mM B-L-L，室温孵育1h，使ClpP1/ClpP2形成有蛋白酶活性的ClpP1P2异源十四聚体形式。
2）反应体系：向96孔板中依次加入反应缓冲液（0.1M KCl，50mM K2HPO4/ KH2PO4 pH7.6, 5mM MgCl2，5%glycerol）、蛋白溶液、B-L-L以及小分子，最后加入作用底物，反应开始。硼替佐米（Bortezomib）作为阳性对照药，用DMSO溶解为1mM母液，受试药物分别设置100μM 、50μM、25μM、10μM、5μM、2.5μM六个作用浓度，反应体系中DMSO含量控制在3%以内。
3）置于30℃孵育，分别于不同时间点进行荧光强度测定。激发波长为380nm，发射波长为460nm，数据读取前震荡15s。
实验结果：
由附图1所示，不同浓度的4-氟-5-羟基-2-甲基吲哚类化合物对结核分枝杆菌ClpP1P2酶活的抑制水平不同，其抑制作用与药物浓度成依赖关系，在100μM作用浓度下，两个4-氟-5-羟基-2-甲基吲哚类化合物—西地尼布（cediranib）和布立尼布（brivanib）对酶活的抑制作用最优。西地尼布（cediranib）对结核分枝杆菌ClpP1P2体外酶活的抑制作用IC50约为4μM，布立尼布（brivanib）对结核分枝杆菌ClpP1P2体外酶活的抑制作用IC50约为14.1μM，因此判定两个4-氟-5-羟基-2-甲基吲哚类化合物均为结核分枝杆菌ClpP1P2抑制剂,用于抗菌药物的开发。
实施例2 4-氟-5-羟基-2-甲基吲哚类化合物在耻垢分枝杆菌模型最低抑菌浓度（MIC）测定
药物及试剂：受试药物购于MCE，利奈唑胺购于辉瑞，Alamar Blue kit 购于Thermo Fisher Scientific，Middlebrook 7H9、7H11购于Gibco，ADC及OADC购于BD, DMSO购于Sigma。
受试菌株：耻垢分枝杆菌（Mycobacterium smegmatis mc2155），来源于中国科学院广州生物医药与健康研究院。
仪器：酶标仪（Biotek）、紫外分光光度计及恒温摇床（Thermo）。
培养基配制：7H9-ADC培养基：7H9干粉4.7g；加入900mL水含2mL甘油，121℃灭菌10min，待温度降至40℃时，加入100mLADC，4℃保存备用，注意无菌操作。
耻垢分枝杆菌培养：将耻垢分枝杆菌接种到7H9-ADC液体培养基，置于220 rpm/min ,37℃恒温摇床进行培养，当OD600达0.6左右取出，在振荡器上振荡培养物2-3min，使菌分散至乳糜状。
受试药物准备：阳性对照以及受试药物母液稀释至1mM，溶剂为DMSO。
实验步骤：
1）药物配置：取6个无菌EP管，分别加入灭菌7H9-ADC培养液，第一管中加入400μL，其余均加入200μL；采用二倍稀释法配置不同浓度的利奈唑胺溶液，依次为2μg/mL、0.5μg/mL、0.25μg/mL、0.125μg/mL、0.0625μg/mL、0.0312μg/mL；受试药物分别采用培养液稀释成128μg/mL 、64μg/mL、32μg/mL、16μg/mL、8μg/mL、4μg/mL；将稀释后的药液，每孔50μL加入96孔板中，每个浓度设置三个复孔。
2）将试管中的菌液进行OD600计数，稀释至1×105个/mL的浓度进行铺板，每孔加入50μL，边孔加入200μL生理盐水，然后用封口膜封口后置于37℃，孵育
3）48h后，每孔中加入10μL Alamar blue，继续培养2-4h。
4）记录96孔板颜色变化。
实验结果：
表1 受试化合物对耻垢分枝杆菌体外抑制（MIC）范围
从上表可知，4-氟-5-羟基-2-甲基吲哚类化合物对耻垢分枝杆菌具有一定的抗菌活性，具有一定的开发前景。
实施例3 4-氟-5-羟基-2-甲基吲哚类化合物对结核分枝杆菌H37Rv抗菌活性及最低抑菌浓度（MIC）测定
药物及试剂：受试药物购于MEC化合物，利福平（RIF）购于Amerco，Middlebrook7H9、7H11购于Gibco，ADC及OADC购于BD。
受试菌株：结核分枝杆菌株（H37Rv，ATCC27294）(委托同济大学附属上海市肺科医院)。
仪器：摇床，二氧化碳培养箱（Thermo）。
实验步骤：
（1）受试菌株的制备：将受试菌株转入液体培养基，经活化后，于37℃培养2周，吸取培养菌液少许，置于4 mL液体培养基中，加入直径3mm无菌玻璃珠20粒，振荡30s，静止沉淀20min，吸取菌悬液上清，用液体培养基调整比浊至1个麦氏单位，相当于1×107CFU/mL备用。
（2）受试化合物的准备：药物用适量DMSO溶解至1mg/mL，0.22μm滤器过滤。再以液体培养基稀释至所需实验浓度（2×终浓度）。受试化合物终浓度设置如下：0.0625μg/mL、0.125μg/mL、0.25μg/mL、0.5μg/mL、1μg/mL、2μg/mL、4μg/mL、8μg/mL、16μg/mL、32μg/mL、64μg/mL，共11个浓度梯度。对照药物利奈唑胺设置浓度为：0.0039μg/mL、0.0078μg/mL、0.0156μg/mL、0.03125μg/mL、0.0625μg/mL、0.125μg/mL、0.25μg/mL、0.5μg/mL、1μg/mL、2μg/mL、4μg/mL，共11个浓度梯度。
（3）操作步骤：检测时，各取上述化合物溶液100μL，加到96孔微孔板中，再加入104CFU/mL（由107 CFU/mL稀释获得）浓度的菌液100μL，使化合物浓度达到2）设置的终浓度。37℃培养，空白对照组不加任何药物，并接种100%、10%、5%和1%的菌量。同一化合物稀释度设两组组平行对照。观察药物对结核分枝杆菌的最低抑菌浓度（MIC）。
实验结果：
表2 化合物对结核分枝杆菌的体外抑菌作用
| 序号 | 化合物 | 分子量 | 结核分枝杆菌MIC（μg/mL） |
| 1 | 利奈唑胺 | 337.35 | 0.0625 |
| 2 | 西地尼布 | 450.5 | 16 |
实施例4 4-氟-5-羟基-2-甲基吲哚类化合物对表皮葡萄球菌最低抑菌浓度（MIC）测定
受试菌株：表皮葡萄球菌耐甲氧西林菌株（B27和ATCC12228）
药物及试剂：受试药物购于MCE，利奈唑胺购于辉瑞公司，MHA、MHB培养基购于BD。
仪器：多点接种仪
药物溶液准备：阳性对照药物利奈唑胺用DMSO配置成1mg/mL的溶液，受试化合物用DMSO配置成10mM的母液。
实验步骤 ：
采用CLSI推荐的标准琼脂二倍稀释法，从MHA平板挑取单克隆菌于3 mL MHB中，过夜培养。取过夜菌液测OD600，稀释至3 mL MHB中（OD=0.05），37 ℃ 220 rpm/min摇菌至对数生长期（0.4-0.8 OD600），测OD600，调整菌液至1×105 CFU/mL备用。利用多点接种仪，涂板于系列倍比稀释的琼脂平板，37℃静置培养18 h，细菌生长的最小浓度即MIC。
实验结果：
表3 4-氟-5-羟基-2-甲基吲哚类化合物对表皮葡萄球菌体外生长抑制MIC的测定
| 序号 | 化合物 | 分子量 | 表皮葡萄球菌MIC（μg/mL） |
| 1 | 利奈唑胺 | 337.4 | 1 |
| 2 | 利福平 | 822.9 | 0.0625 |
| 3 | 西地尼布 | 450.5 | 64 |
| 4 | 布立尼布 | 441.5 | / |
从表3可知，4-氟-5-羟基-2-甲基吲哚类化合物对表皮葡萄球菌表现出一定的抑制作用，具有一定的开发前景。
Claims (1)
1.一种结构式如通式I所示的4-氟-5-羟基-2-甲基吲哚类化合物在制备抗菌药物中的用途，其特征在于，所述化合物的结构式中包含4-氟-2-甲基吲哚结构单元，并可抑制革兰氏阳性菌的生长；所述革兰氏阳性菌为结核分枝杆菌、 耻垢分枝杆菌和表皮葡萄球菌；
其中R为
当R为
时，式I所示分子即为西地尼布(cediranib)；
当R为
时，式I所示分子为布立尼布(brivanib)。
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201811330994.8A CN111166750B (zh) | 2018-11-09 | 2018-11-09 | 一种4-氟-2-甲基吲哚类化合物的抗菌新用途 |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201811330994.8A CN111166750B (zh) | 2018-11-09 | 2018-11-09 | 一种4-氟-2-甲基吲哚类化合物的抗菌新用途 |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN111166750A CN111166750A (zh) | 2020-05-19 |
| CN111166750B true CN111166750B (zh) | 2022-12-27 |
Family
ID=70617303
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201811330994.8A Active CN111166750B (zh) | 2018-11-09 | 2018-11-09 | 一种4-氟-2-甲基吲哚类化合物的抗菌新用途 |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN111166750B (zh) |
Family Cites Families (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20120141491A1 (en) * | 2009-08-11 | 2012-06-07 | University Of Florida Research Foundation Inc. | Methods and compositions for the treatment of cancers and pathogenic infections |
| SG180031A1 (en) * | 2010-10-15 | 2012-05-30 | Agency Science Tech & Res | Combination treatment of cancer |
| CN102516113B (zh) * | 2011-11-11 | 2014-04-09 | 四川大学 | 系列联苯醚衍生物以及在制备抗结核病药物方面的用途 |
| CN105026397B (zh) * | 2013-03-05 | 2017-06-30 | 默克专利股份公司 | 作为抗癌剂的9‑(芳基或杂芳基)‑2‑(吡唑基、吡咯烷基或环戊基)氨基嘌呤衍生物 |
| WO2015037005A1 (en) * | 2013-09-11 | 2015-03-19 | Compugen Ltd. | Anti-vstm5 antibodies and the use thereof in therapy and diagnosis |
| CN108420814A (zh) * | 2017-02-15 | 2018-08-21 | 四川大学 | 一种nps-2143用于抗菌的新用途 |
| CN108420822B (zh) * | 2017-02-15 | 2020-04-17 | 四川大学 | 色瑞替尼的抗菌新用途 |
| CN110563656A (zh) * | 2018-06-06 | 2019-12-13 | 四川大学 | 嘧啶类小分子化合物及在制备抗分枝杆菌药物中的用途 |
-
2018
- 2018-11-09 CN CN201811330994.8A patent/CN111166750B/zh active Active
Also Published As
| Publication number | Publication date |
|---|---|
| CN111166750A (zh) | 2020-05-19 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Barry et al. | An improved single-disk method for testing the antibiotic susceptibility of rapidly-growing pathogens | |
| Casewell et al. | In-vitro activity of mupirocin (‘pseudomonic acid’) against clinical isolates of Staphylococcus aureus | |
| Chakraborty et al. | Biochemical characters and antibiotic susceptibility of Staphylococcus aureus isolates | |
| Ojo et al. | Plasmid Curing Analysis of Antibiotic Resistance in [3-lactamase Producing Staphylococci from Wounds and Burns Patients | |
| Al-Jumaily et al. | Multidrug resistant Proteus mirabilis isolated from urinary tract infection from different hospitals in Baghdad City | |
| Ghatage et al. | Antimicrobial screening | |
| CN108420822B (zh) | 色瑞替尼的抗菌新用途 | |
| CN111166750B (zh) | 一种4-氟-2-甲基吲哚类化合物的抗菌新用途 | |
| Alsohaili et al. | Bacterial pathogen in urinary tract infection and antibiotic resistance patteern in Zaraqa-Jordan | |
| RU2319746C2 (ru) | Способ ускоренного определения чувствительности буркхольдерий к химиопрепаратам | |
| CN111979292B (zh) | 一种同时携带多重耐药基因cfr和lsa(E)的MRSA的用途 | |
| KR101824588B1 (ko) | 메티실린-저항성 ｓ. 아우레우스의 스크리닝 또는 농축을 위한 배양 배지 | |
| CN110804646B (zh) | 一种用于利福平胶囊微生物限度控制菌检查的抗菌活性去除方法 | |
| CN103550217B (zh) | 一种靶向于细菌rna聚合酶的抗结核小分子化合物 | |
| Mustafa et al. | Phenotypic detection of virulence determinants and antibiotics resistance in Staphylococcus aureus from different clinical isolates in Kirkuk city | |
| CN113262214A (zh) | 盐酸西那卡塞在制备抗菌药物中的应用 | |
| RU2788607C1 (ru) | Способ получения питательной среды для селективного выявления Candida auris | |
| US20240132933A1 (en) | Improved anti-biofilm assay methods | |
| CN102370633A (zh) | 一种细菌YycG组氨酸激酶抑制剂 | |
| RU2266956C2 (ru) | Питательная среда для выделения бруцелл | |
| Fatima | Isolation of antibiotic producing bacteria from soil and identification by 16S rRNA gene sequencing | |
| Nureen et al. | Evaluation of bacterial culture and their resistant pattern in pus containing patients of different wards of the hospital, Lahore, Pakistan | |
| Scott et al. | CLINICAL STUDY UPDATE ON A NOVEL RIBOSOMAL RNA-BASED RAPID DIAGNOSTIC METHOD TO DETECT, IDENTIFY AND ASSESS ANTIBIOTIC SUSCEPTIBILITY OF UROPATHOGENS: MP23-18 | |
| Afolabi et al. | ASSESSMENT OF ANTIMICROBIAL ACTIVITY OF PLECTASIN AGAINST LOCAL STRAINS OF GRAM NEGATIVEANDPOSITIVEBACTERIA | |
| Klochko et al. | Antimicrobial and cytotoxic characteristicsof antibiotic streptofungin |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |