CN112076182B - Dna拓扑异构酶抑制剂在制备预防和/或治疗新型冠状病毒感染药物中的应用 - Google Patents
Dna拓扑异构酶抑制剂在制备预防和/或治疗新型冠状病毒感染药物中的应用 Download PDFInfo
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Abstract
本发明属于抗病毒药物领域，公开了DNA拓扑异构酶抑制剂在制备预防和/或治疗新型冠状病毒感染药物中的应用，所述DNA拓扑异构酶抑制剂包括米托蒽醌、阿霉素或以上小分子化合物药学上可接受的盐。马米托蒽醌、阿霉素等DNA拓扑异构酶抑制剂可极强的抑制新冠病毒SARS‑CoV‑2复制子RNA的合成及降低野生型新冠病毒SARS‑CoV‑2在细胞中的RNA拷贝量，从而抑制了新型冠状病毒SARS‑CoV‑2的复制，具有预防和/或治疗新型冠状病毒SARS‑CoV‑2感染的作用。
Description
技术领域
本发明涉及抗病毒药物领域，尤其涉及DNA拓扑异构酶抑制剂在制备预防和/或治疗新型冠状病毒感染药物中的应用。
背景技术
自新型冠状病毒病COVID-19于2019年底爆发以来，该疾病已在全球迅速蔓延，造成巨大的全球经济下滑和医疗负担。截止2020年8月5日，根据美国约翰霍普金斯大学的数据显示，全球累计报告确证病例18,543,662例，累计死亡病例700,714例。
目前，2019冠状病毒病尚无特效药，目前治疗多为对症治疗，无法有效逆转重症患者病程进展。新型冠状病毒与SARS-CoV以及MERS-CoV虽有部分相似，但截然不同，部分患者在经过抗病毒和抗炎治疗后效果不佳，容易合并感染，造成多发性，弥漫性病变。因此针对新型冠状病毒的靶点开展靶向药物研究具有重大意义。
DNA拓扑异构酶抑制剂是一类能够抑制DNA拓扑异构酶活性的化合物一类能够抑制DNA拓扑异构酶活性的化合物，常见的DNA拓扑异构酶抑制剂包括米托蒽醌(Mitoxantrone)、阿霉素(Adriamycin)等DNA拓扑异构酶II抑制剂。
其中，米托蒽醌(Mitoxantrone)是一种type II topoisomerase和proteinkinase C(PKC)的抑制剂，对于PKC的IC50值为8.5μM。Mitoxantrone可抑制MCF-7/wt cells的细胞增殖，对应的IC50值为0.42μM。Mitoxantrone还可诱导细胞凋亡。米托蒽醌的结构式如下所示：
阿霉素(Adriamycin)是一种抗生素类试剂，可抑制DNA topoisomerase II，并在肿瘤细胞中诱导DNA损伤、线粒体自噬和凋亡。Doxorubicin可降低AMPK的基础磷酸化。Doxorubicin可应用于HIV感染病人的联合治疗，但是在免疫治疗中有将HBV重活化的风险。阿霉素的结构式如下所示：
以上各种DNA拓扑异构酶抑制剂对多种疾病中具有良好的效果，但是目前尚无将DNA拓扑异构酶抑制剂应用于预防和/或治疗新型冠状病毒感染的相关报道。
发明内容
本发明针对目前由新型冠状病毒SARS-CoV-2引起的感染缺乏有效防治药物的问题，提供米托蒽醌、阿霉素等DNA拓扑异构酶抑制剂或以上小分子化合物药学上可接受的盐在制备预防和/或治疗新型冠状病毒SARS-CoV-2感染药物中的应用。本发明利用新型冠状病毒SARS-CoV-2安全型复制子，及分离出的野生型新型冠状病毒SARS-CoV-2，通过细胞实验证实了米托蒽醌、阿霉素等DNA拓扑异构酶抑制剂可抑制病毒的RNA合成，阻止病毒的转录复制。可见米托蒽醌、阿霉素等DNA拓扑异构酶抑制剂具有预防和/或治疗新型冠状病毒感染的作用，可制备成为预防和/或治疗新型冠状病毒炎症药物进行应用。
本发明的第二个目的是提供DNA拓扑异构酶抑制剂在制备预防和/或治疗新型冠状病毒炎症药物中的应用。
本发明的第三个目的是提供DNA拓扑异构酶抑制剂在制备新型冠状病毒RNA合成抑制剂中的应用。
本发明的第四个目的是提供一种预防和/或治疗新型冠状病毒感染的药物。
本发明的上述目的是通过以下方案予以实现的：
DNA拓扑异构酶抑制剂在制备预防和/或治疗新型冠状病毒感染药物中的应用。
进一步，所述DNA拓扑异构酶抑制剂选自米托蒽醌、阿霉素及其药学上可接受的盐的至少一种。
进一步，所述新型冠状病毒为2019新型冠状病毒(2019-nCoV，即SARS-CoV-2)。
DNA拓扑异构酶抑制剂在制备预防和/或治疗新型冠状病毒炎症药物中的应用。
进一步，所述DNA拓扑异构酶抑制剂选自米托蒽醌、阿霉素及其药学上可接受的盐的至少一种。
进一步，所述新型冠状病毒炎症为新型冠状病毒肺炎(COVID-19)。
DNA拓扑异构酶抑制剂在制备新型冠状病毒RNA复制抑制剂中的应用。
进一步，所述新型冠状病毒为2019新型冠状病毒(2019-nCoV，即SARS-CoV-2)。
一种预防和/或治疗新型冠状病毒感染的药物，包括DNA拓扑异构酶抑制剂。
进一步，所述DNA拓扑异构酶抑制剂选自米托蒽醌、阿霉素及其药学上可接受的盐的至少一种。
进一步，所述药物为胶囊、片剂、丸剂、颗粒剂、冲剂、注射药剂或喷剂。
进一步，所述药物还可以包括药学上可接受的辅料或载体。
进一步，所述药物还可以包含其他抗冠状病毒药物。
进一步，所述药物还可以包含增强人体免疫能力的成分。
当所述米托蒽醌，阿霉素及其以上药物药学上可接受的盐抑制新型冠状病毒复制的同时，采用其他抗冠状病毒抑制已有新型冠状病毒，并增加人体的免疫力，增强人体自身对新型冠状病毒的抑制和清除，从而达到清除新型冠状病毒的目的。
与现有技术相比，本发明具有以下有益效果：
本发明首次将DNA拓扑异构酶抑制剂应用于预防和/或治疗新型冠状病毒感染，米托蒽醌、阿霉素等DNA拓扑异构酶抑制剂能较强的抑制新型冠状病毒RNA的转录和复制，从而抑制了新型冠状病毒的复制，因此所述米托蒽醌、阿霉素等DNA拓扑异构酶抑制剂或其药学上可接受的盐具有预防和/或防治新型冠状病毒感染的作用，可制备为抗新冠病毒药物进行应用；同时还可以与其他抗冠状病毒药物、增强人体免疫能力的药物共同应用，进一步强化疗效。
附图说明
图1表示米托蒽醌、阿霉素对SARS-CoV-2安全型复制子的RNA合成的抑制作用；
图2表示米托蒽醌、阿霉素对野生型SARS-CoV-2复制的抑制作用。
具体实施方式
下面结合说明书附图和具体实施例对本发明作出进一步地详细阐述，所述实施例只用于解释本发明，并非用于限定本发明的范围。下述实施例中所使用的试验方法如无特殊说明，均为常规方法；所使用的材料、试剂等，如无特殊说明，为可从商业途径得到的试剂和材料。
实施例1
SARS-CoV-2安全型复制子可以模拟SARS-CoV-2在细胞中的RNA转录和复制，细胞中的荧光素酶活性可以直接反应SARS-CoV-2的RNA转录和复制程度，同时不会产生出具有感染性的新型冠状病毒SARS-CoV-2。因此本发明利用新型冠状病毒SARS-CoV-2安全型复制子验证米托蒽醌、阿霉素对SARS-CoV-2的RNA合成的抑制作用。
相关实验可以在P2实验室中进行，具体实验步骤如下：
将SARS-CoV-2安全型复制子构成质粒ps2V(SEQ ID NO：1)、ps2AN(SEQ ID NO：2)、ps2AC(SEQ ID NO：3)、ps2B(SEQ ID NO：4)，按照一定比例用Thermo Fisher公司的Lipo2000转染试剂及Opti-MEM转染到提前一天种植在用多聚赖氨酸处理的12孔细胞上HEK293T细胞中(细胞密度约为6.5×104/cm2)，试剂用量见下表1。具体地，使用100μL Opti-MEM溶解质粒，100μLOpti-MEM溶解Lipo 2000，分别孵育5min后混合，继续孵育20min后加入HEK293T细胞。
表1.Opti-Lipo2000-DNA混合液体系
转染6h后，按照浓度梯度(20μM，10μM，5μM，2.5μM，1.25μM，0.625μM，0.3125μM，0.15625μM，0.078125μM，0.0390625μM)加入米托蒽醌或阿霉素。药物处理24h后，检测细胞荧光素酶活性，以DMSO对照为基准，计算抑制率，并利用Graphpad Prism 7.0软件统计药物的半抑制浓度(下称IC50)。每个浓度下重复3组实验，所得数据如下表2所示，对表2的数据进行处理得到图1所示曲线。
表2.不同浓度下米托蒽醌、阿霉素的抑制率
图1和表2结果显示，米托蒽醌、阿霉素对SARS-CoV-2的RNA合成具有抑制作用，其中米托蒽醌的IC50为0.3378±0.2348μM，阿霉素的IC50为0.8244±0.1136μM，说明米托蒽醌、阿霉素可抑制新型冠状病毒RNA的转录和复制，对新型冠状病毒的感染有治疗作用。
实施例2
本发明利用分离出的野生型新型冠状病毒SARS-CoV-2验证米托蒽醌、阿霉素对病毒复制的抑制作用，具体实验方法如下：
将生长状态良好的HEK293T细胞平均铺于用多聚赖氨酸处理的48孔培养板中(细胞密度约为6.5×104/cm2)。当细胞生长16h后(细胞密度约为1.6×105/mL)，转染从北京义翘神州公司购买的表达SARS-CoV-2受体ACE2的pCMV-ACE2-FLAG质粒0.1μg。转染后24h，用PBS冲洗细胞后，感染野生型SARS-CoV-2(MOI＝0.1，37℃，1h)。随后，替换含有不同浓度梯度(20μM，5μM，1.25μM，0.3125μM，0.078125μM，0.01953125μM)的米托蒽醌和阿霉素的DMEM(2％FBS)。药物处理24h后，利用TRIZOL提取细胞RNA。SARS-CoV-2的RNA拷贝利用达安基因的新型冠状病毒2019-nCOV核酸检测(PCR-荧光探针法)进行检测，获得Ct值，根据标准曲线计算病毒拷贝数，计算抑制率，并利用Graphpad Prism 7.0软件统计药物的IC50。每个浓度下重复3组实验，所得数据如下表3所示，对表3的数据进行处理得到图2所示曲线。
表3.不同浓度下米托蒽醌、阿霉素的抑制率
根据图2和表3的测试结果可以看出，米托蒽醌、阿霉素能够降低野生型新冠病毒SARS-CoV-2在细胞中的RNA拷贝量，抑制野生型SARS-CoV-2生长。在抑制野生型SARS-CoV-2生长时，米托蒽醌的IC50为0.1333±0.0901μM，阿霉素的IC50为0.6117±0.2282μM。以上实验结果进一步说明米托蒽醌、阿霉素可抑制新型冠状病毒RNA的转录和复制，对新型冠状病毒的感染有治疗作用。
SEQUENCE LISTING
<110> 中山大学
<120> DNA拓扑异构酶抑制剂在制备预防和/或治疗新型冠状病毒感染药物中的应用
<130>
<160> 4
<170> PatentIn version 3.5
<210> 1
<211> 4364
<212> DNA
<213> 人工序列
<400> 1
gctagcatta aaggtttata ccttcccagg taacaaacca accaactttc gatctcttgt 60
agatctgttc tctaaacgaa ctttaaaatc tgtgtggctg tcactcggct gcatgcttag 120
tgcactcacg cagtataatt aataactaat tactgtcgtt gacaggacac gagtaactcg 180
tctatcttct gcaggctgct tacggtttcg tccgtgttgc agccgatcat cagcacatct 240
aggtttcgtc cgggtgtgac cgaaaggtaa ggtggagagc cttgtccctg gtttcaacga 300
gaaaacacac gtccaactca gtttgcctgt tttacaggtt cgcgacgtgc tcgtacgtgg 360
ctttggagac tccgtggagg aggtcttatc agaggcacgt caacatctta aagatggcac 420
ttgtggctta gtagaagttg aaaaaggcgt tttgcctcaa cttgaacagc ctgagctttg 480
ggctaagcgc aacattaaac cagtaccaga ggtgaaaata ctcaataatt tgggtgtgga 540
cattgctgct aatactgtga tctgggacta caaaagagat gctccagcac atatatctac 600
tattggtgtt tgttctatga ctgacatagc caagaaacca actgaaacga tttgtgcacc 660
actcactgtc ttttttgatg gtagagttga tggtcaagta gacttattta gaaatgcccg 720
taatggtgtt cttattacag aaggtagtgt taaaggttta caaccatctg taggtcccaa 780
acaagctagt cttaatggag tcacattaat tggagaagcc gtaaaaacac agttcaatta 840
ttataagaaa gttgatggtg ttgtccaaca attacctgaa acttacttta ctcagagtag 900
aaatttacaa gaatttaaac ccaggagtca aatggaaatt gatttcttag aattagctat 960
ggatgaattc attgaacggt ataaattaga aggctatgcc ttcgaacata tcgtttatgg 1020
agattttagt catgagggcc cggaaacctg gccctgtctt cttgacgagc attcctaggg 1080
gtctttcccc tctcgccaaa ggaatgcaag gtctgttgaa tgtcgtgaag gaagcagttc 1140
ctctggaagc ttcttgaaga caaacaacgt ctgtagcgac cctttgcagg cagcggaacc 1200
ccccacctgg cgacaggtgc ctctgcggcc aaaagccacg tgtataagat acacctgcaa 1260
aggcggcaca accccagtgc cacgttgtga gttggatagt tgtggaaaga gtcaaatggc 1320
tctcctcaag cgtattcaac aaggggctga aggatgccca gaaggtaccc cattgtatgg 1380
gatctgatct ggggcctcgg tgcacatgct ttacatgtgt ttagtcgagg ttaaaaaaac 1440
gtctaggccc cccgaaccac ggggacgtgg ttttcctttg aaaaacacga tgataagcgg 1500
ccgcatggtg agcaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct 1560
ggacggcgac gtaaacggcc acaagttcag cgtgtccggc gagggcgagg gcgatgccac 1620
ctacggcaag ctgaccctga agttcatctg caccaccggc aagctgcccg tgccctggcc 1680
caccctcgtg accaccctga cctacggcgt gcagtgcttc agccgctacc ccgaccacat 1740
gaagcagcac gacttcttca agtccgccat gcccgaaggc tacgtccagg agcgcaccat 1800
cttcttcaag gacgacggca actacaagac ccgcgccgag gtgaagttcg agggcgacac 1860
cctggtgaac cgcatcgagc tgaagggcat cgacttcaag gaggacggca acatcctggg 1920
gcacaagctg gagtacaact acaacagcca caacgtctat atcatggccg acaagcagaa 1980
gaacggcatc aaggtgaact tcaagatccg ccacaacatc gaggacggca gcgtgcagct 2040
cgccgaccac taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacaa 2100
ccactacctg agcacccagt ccgccctgag caaagacccc aacgagaagc gcgatcacat 2160
ggtcctgctg gagttcgtga ccgccgccgg gatcactctc ggcatggacg agctgtacaa 2220
gtaataataa taagatatct gatcttctgg tctaaacgaa ctaaatatta tattagtttt 2280
tctgtttgga actttaattt tagccatggc cgatgctaag aacattaaga agggccctgc 2340
tcccttctac cctctggagg atggcaccgc tggcgagcag ctgcacaagg ccatgaagag 2400
gtatgccctg gtgcctggca ccattgcctt caccgatgcc cacattgagg tggacatcac 2460
ctatgccgag tacttcgaga tgtctgtgcg cctggccgag gccatgaaga ggtacggcct 2520
gaacaccaac caccgcatcg tggtgtgctc tgagaactct ctgcagttct tcatgccagt 2580
gctgggcgcc ctgttcatcg gagtggccgt ggcccctgct aacgacattt acaacgagcg 2640
cgagctgctg aacagcatgg gcatttctca gcctaccgtg gtgttcgtgt ctaagaaggg 2700
cctgcagaag atcctgaacg tgcagaagaa gctgcctatc atccagaaga tcatcatcat 2760
ggactctaag accgactacc agggcttcca gagcatgtac acattcgtga catctcatct 2820
gcctcctggc ttcaacgagt acgacttcgt gccagagtct ttcgacaggg acaaaaccat 2880
tgccctgatc atgaacagct ctgggtctac cggcctgcct aagggcgtgg ccctgcctca 2940
tcgcaccgcc tgtgtgcgct tctctcacgc ccgcgaccct attttcggca accagatcat 3000
ccccgacacc gctattctga gcgtggtgcc attccaccac ggcttcggca tgttcaccac 3060
cctgggctac ctgatttgcg gctttcgggt ggtgctgatg taccgcttcg aggaggagct 3120
gttcctgcgc agcctgcaag actacaaaat tcagtctgcc ctgctggtgc caaccctgtt 3180
cagcttcttc gctaagagca ccctgatcga caagtacgac ctgtctaacc tgcacgagat 3240
tgcctctggc ggcgccccac tgtctaagga ggtgggcgaa gccgtggcca agcgctttca 3300
tctgccaggc atccgccagg gctacggcct gaccgagaca accagcgcca ttctgattac 3360
cccagagggc gacgacaagc ctggcgccgt gggcaaggtg gtgccattct tcgaggccaa 3420
ggtggtggac ctggacaccg gcaagaccct gggagtgaac cagcgcggcg agctgtgtgt 3480
gcgcggccct atgattatgt ccggctacgt gaataaccct gaggccacaa acgccctgat 3540
cgacaaggac ggctggctgc actctggcga cattgcctac tgggacgagg acgagcactt 3600
cttcatcgtg gaccgcctga agtctctgat caagtacaag ggctaccagg tggccccagc 3660
cgagctggag tctatcctgc tgcagcaccc taacattttc gacgccggag tggccggcct 3720
gcccgacgac gatgccggcg agctgcctgc cgccgtcgtc gtgctggaac acggcaagac 3780
catgaccgag aaggagatcg tggactatgt ggccagccag gtgacaaccg ccaagaagct 3840
gcgcggcgga gtggtgttcg tggacgaggt gcccaagggc ctgaccggca agctggacgc 3900
ccgcaagatc cgcgagatcc tgatcaaggc taagaaaggc ggcaagatcg ccgtgtaagg 3960
atccgtgggc tatataaacg ttttcgcttt tccgtttacg atatatagtc tactcttgtg 4020
cagaatgaat tctcgtaact acatagcaca agtagatgta gttaacttta atctcacata 4080
gcaatcttta atcagtgtgt aacattaggg aggacttgaa agagccacca cattttcacc 4140
gaggccacgc ggagtacgat cgagtgtaca gtgaacaatg ctagggagag ctgcctatat 4200
ggaagagccc taatgtgtaa aattaatttt agtagtgcta tccccatgtg attttaatag 4260
cttcttagga gaatgacaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa ctagcataac 4320
cccttggggc ctctaaacgg gtcttgaggg gttttttgtc taga 4364
<210> 2
<211> 10429
<212> DNA
<213> 人工序列
<400> 2
gctagcgagg gcccggaaac ctggccctgt cttcttgacg agcattccta ggggtctttc 60
ccctctcgcc aaaggaatgc aaggtctgtt gaatgtcgtg aaggaagcag ttcctctgga 120
agcttcttga agacaaacaa cgtctgtagc gaccctttgc aggcagcgga accccccacc 180
tggcgacagg tgcctctgcg gccaaaagcc acgtgtataa gatacacctg caaaggcggc 240
acaaccccag tgccacgttg tgagttggat agttgtggaa agagtcaaat ggctctcctc 300
aagcgtattc aacaaggggc tgaaggatgc ccagaaggta ccccattgta tgggatctga 360
tctggggcct cggtgcacat gctttacatg tgtttagtcg aggttaaaaa aacgtctagg 420
ccccccgaac cacggggacg tggttttcct ttgaaaaaca cgatgataaa tggagtccct 480
ggtgcccggc ttcaacgaga agacccacgt gcagctgtct ctgcctgtgc tgcaggtgag 540
ggatgtgctg gtgcgcggct ttggcgactc cgtcgaggag gtgctgtctg aggccaggca 600
gcacctgaag gacggaacct gcggactggt ggaggtggag aagggcgtgc tgccacagct 660
ggagcagcct tacgtgttca tcaagaggtc cgatgcaagg acagcaccac acggacacgt 720
gatggtggag ctggtggccg agctggaggg catccagtat ggccgctctg gagagaccct 780
gggcgtgctg gtgccacacg tgggagagat cccagtggcc tatcggaagg tgctgctgag 840
aaagaacggc aataagggag caggaggaca ctcttacgga gcagacctga agagcttcga 900
tctgggcgac gagctgggca ccgatcctta tgaggacttt caggagaact ggaatacaaa 960
gcacagctcc ggcgtgaccc gggagctgat gagagagctg aacggcggcg cctacaccag 1020
atatgtggat aacaatttct gcggaccaga cggatacccc ctggagtgta tcaaggatct 1080
gctggccaga gcaggcaagg cctcctgcac cctgtctgag cagctggact tcatcgacac 1140
aaagcggggc gtgtattgct gtagagagca cgagcacgag atcgcctggt ataccgagcg 1200
gtccgagaag tcttacgagc tgcagacacc attcgagatc aagctggcca agaagttcga 1260
caccttcaac ggcgagtgtc caaacttcgt gtttcccctg aatagcatca tcaagaccat 1320
ccagcccaga gtggagaaga agaagctgga tggctttatg ggcaggatcc gcagcgtgta 1380
ccctgtggcc tccccaaacg agtgcaatca gatgtgcctg tccacactga tgaagtgcga 1440
tcactgtggc gagacctctt ggcagacagg cgacttcgtg aaggccacct gcgagttttg 1500
tggcaccgag aacctgacaa aggagggcgc caccacatgc ggctatctgc ctcagaatgc 1560
cgtggtgaag atctactgcc cagcctgtca caactccgaa gtgggaccag agcactctct 1620
ggccgagtac cacaatgagt ccggcctgaa gacaatcctg aggaagggag gaaggaccat 1680
cgccttcggc ggatgcgtgt tttcttatgt gggctgccac aacaagtgtg catactgggt 1740
gccaagggcc agcgccaata tcggctgtaa ccacaccgga gtggtgggag agggatccga 1800
gggcctgaac gataatctgc tggagatcct gcagaaggag aaggtgaaca tcaatatcgt 1860
gggcgacttc aagctgaacg aggagatcgc catcatcctg gcctccttct ctgccagcac 1920
atccgccttt gtggagaccg tgaagggcct ggactacaag gccttcaagc agatcgtgga 1980
gagctgcggc aacttcaagg tgaccaaggg caaggccaag aagggcgcct ggaacatcgg 2040
cgagcagaag agcatcctgt cccctctgta tgccttcgcc agcgaggcag caagggtggt 2100
gagatctatc tttagccgga ccctggagac agcccagaat tccgtgagag tgctgcagaa 2160
ggccgccatc accatcctgg atggcatctc ccagtactct ctgaggctga tcgatgccat 2220
gatgttcacc tccgacctgg ccacaaacaa tctggtggtc atggcctaca tcaccggcgg 2280
cgtggtgcag ctgacctctc agtggctgac aaacatcttt ggcaccgtgt atgagaagct 2340
gaagccagtg ctggattggc tggaggagaa gttcaaggag ggcgtggagt ttctgcgcga 2400
cggctgggag atcgtgaagt tcatcagcac ctgcgcatgt gagatcgtgg gaggacagat 2460
cgtgacctgt gccaaggaga tcaaggagtc cgtgcagaca ttctttaagc tggtgaacaa 2520
gttcctggcc ctgtgcgccg actctatcat catcggcggc gccaagctga aggccctgaa 2580
cctgggcgag acctttgtga cacacagcaa gggcctgtac aggaagtgcg tgaagtcccg 2640
cgaggagacc ggactgctga tgcccctgaa ggcacctaag gagatcatct tcctggaggg 2700
cgagaccctg cccacagagg tgctgacaga ggaggtggtg ctgaagaccg gcgacctgca 2760
gccactggag cagcccacca gcgaggcagt ggaggcacct ctggtgggca caccagtgtg 2820
catcaatggc ctgatgctgc tggagatcaa ggataccgag aagtactgtg ccctggcccc 2880
taacatgatg gtgacaaaca ataccttcac actgaagggc ggcgccccaa ccaaggtgac 2940
atttggcgac gataccgtga tcgaggtgca gggctacaag tctgtgaata tcacattcga 3000
gctggatgag agaatcgaca aggtgctgaa cgagaagtgc agcgcctata cagtggagct 3060
gggcaccgag gtgaacgagt ttgcctgcgt ggtggccgac gccgtgatca agaccctgca 3120
gccagtgtcc gagctgctga cacccctggg catcgatctg gacgagtggt ctatggccac 3180
ctactatctg ttcgacgaga gcggcgagtt taagctggcc tcccacatgt actgctcttt 3240
ctatccccct gatgaagacg aggaggaggg cgattgcgag gaggaggagt ttgagcccag 3300
cacacagtac gagtatggca ccgaggacga ttaccagggc aagccactgg agttcggagc 3360
cacctccgcc gccctgcagc cagaggagga gcaggaggag gattggctgg acgatgactc 3420
ccagcagacc gtgggccagc aggatggctc tgaggacaat cagaccacaa ccatccagac 3480
aatcgtggag gtgcagcctc agctggagat ggagctgacc ccagtggtgc agaccatcga 3540
ggtgaactct ttcagcggct atctgaagct gacagataac gtgtacatca agaacgccga 3600
cattgtggag gaggccaaga aggtgaagcc taccgtggtg gtgaacgccg ccaacgtgta 3660
cctgaagcac ggaggaggag tggcaggcgc cctgaacaag gccaccaaca atgccatgca 3720
ggtggagagc gatgactata tcgccacaaa tggacccctg aaggtcggag gaagctgcgt 3780
gctgtccgga cacaacctgg ccaagcactg tctgcacgtg gtgggcccta acgtgaataa 3840
gggcgaggac atccagctgc tgaagtccgc ctacgagaac ttcaatcagc acgaggtgct 3900
gctggcccct ctgctgagcg ccggcatctt tggcgccgat ccaatccact ccctgagggt 3960
gtgcgtggac accgtgcgca caaacgtgta cctggccgtg ttcgataaga acctgtacga 4020
caagctggtg tctagctttc tggagatgaa gagcgagaag caggtggagc agaagatcgc 4080
cgagatccct aaggaggagg tgaagccatt catcaccgag agcaagcctt ccgtggagca 4140
gaggaagcag gatgacaaga agatcaaggc ctgcgtggag gaggtgacaa ccacactgga 4200
ggagaccaag ttcctgacag agaacctgct gctgtacatc gatatcaacg gcaatctgca 4260
cccagacagc gccacactgg tgtccgatat cgacatcacc tttctgaaga aggatgcccc 4320
atatatcgtg ggcgacgtgg tgcaggaggg cgtgctgaca gccgtggtca tccccaccaa 4380
gaaggccggc ggcaccacag agatgctggc caaggccctg cgcaaggtgc ctaccgacaa 4440
ttacatcacc acatatccag gccagggcct gaacggctat accgtggagg aggccaagac 4500
cgtgctgaag aagtgcaaga gcgccttcta catcctgcct tctatcatca gcaatgagaa 4560
gcaggagatc ctgggcaccg tgtcctggaa cctgagggag atgctggccc acgccgagga 4620
gacacgcaag ctgatgcccg tgtgcgtgga gacaaaggcc atcgtgagca ccatccagcg 4680
gaagtataag ggcatcaaga tccaggaggg agtggtggac tacggagcaa gattctactt 4740
ttatacctct aagaccacag tggccagcct gatcaacaca ctgaatgatc tgaacgagac 4800
cctggtgaca atgcccctgg gctatgtgac ccacggcctg aatctggagg aggccgccag 4860
gtacatgcgc tccctgaagg tgccagcaac cgtgagcgtg agctctcctg acgccgtgac 4920
agcctacaac ggctatctga caagctcctc taagacccca gaggagcact tcatcgagac 4980
catctctctg gccggcagct ataaggattg gtcctactct ggccagtcca cacagctggg 5040
catcgagttt ctgaagaggg gcgacaagag cgtgtactat accagcaatc ccaccacatt 5100
ccacctggat ggcgaagtga tcaccttcga caacctgaag accctgctga gcctgcggga 5160
ggtgagaacc atcaaggtgt tcaccacagt ggataacatc aatctgcaca cacaggtggt 5220
ggacatgtcc atgacctatg gccagcagtt tggcccaaca tacctggatg gcgccgacgt 5280
gaccaagatc aagccccaca atagccacga gggcaagaca ttctacgtgc tgcctaatgc 5340
caccaacttt tccctgctga agcaggcagg cgacgtggag gagaacccag gaccagatga 5400
caccctgagg gtggaggcct tcgagtacta tcacaccaca gatcctagct ttctgggccg 5460
ctatatgtcc gccctgaatc acaccaagaa gtggaagtac ccacaggtga acggcctgac 5520
aagcatcaag tgggccgaca acaattgcta cctggccacc gccctgctga cactgcagca 5580
gatcgagctg aagttcaacc cacccgccct gcaggatgca tactataggg caagagcagg 5640
agaggcagcc aatttttgcg ccctgatcct ggcctattgt aacaagaccg tgggagagct 5700
gggcgatgtg cgggagacaa tgagctacct gttccagcac gccaatctgg actcctgcaa 5760
gagagtgctg aacgtggtgt gcaagacatg tggccagcag cagaccacac tgaagggcgt 5820
ggaggccgtg atgtatatgg gcaccctgag ctacgagcag tttaagaagg gcgtgcagat 5880
cccctgcaca tgtggcaagc aggccaccaa gtacctggtg cagcaggagt cccctttcgt 5940
gatgatgtct gcccctccag cccagtatga gctgaagcac ggcaccttta catgcgcctc 6000
tgagtacacc ggcaattatc agtgtggcca ctataagcac atcaccagca aggagacact 6060
gtactgcatc gatggcgccc tgctgaccaa gagctccgag tacaagggcc ccatcacaga 6120
cgtgttctat aaggagaatt cttacaccac aaccatcgcc accaacttta gcctgctgaa 6180
gcaggccggc gatgtggagg agaaccctgg accaaagccc gtgacctata agctggacgg 6240
cgtggtgtgc acagagatcg atcctaagct ggacaactac tacaagaagg ataactctta 6300
tttcaccgag cagcccatcg acctggtgcc taatcagcct tacccaaacg ccagcttcga 6360
taatttcaag ttcgtgtgcg acaatatcaa gtttgccgat gacctgaacc agctgaccgg 6420
atacaagaag ccagccagcc gggagctgaa ggtgacattc tttcctgatc tgaacggcga 6480
cgtggtggcc atcgactaca agcactatac accttccttc aagaagggcg ccaagctgct 6540
gcacaagcca atcgtgtggc acgtgaacaa tgccaccaat aaggccacat acaagccaaa 6600
cacctggtgc atcagatgtc tgtggtctac aaagcccgtg gagaccagca attcctttga 6660
tgtgctgaag agcgaggatg cccagggcat ggacaacctg gcctgcgagg acctgaagcc 6720
cgtgagcgag gaggtggtgg agaatcctac catccagaag gatgtgctgg agtgtaacgt 6780
gaagacaacc gaggtggtgg gcgacatcat cctgaagcct gccaacaatt ccctgaagat 6840
cacagaggaa gtgggccaca ccgatctgat ggccgcctac gtggacaatt ctagcctgac 6900
catcaagaag ccaaacgagc tgagcagggt gctgggcctg aagaccctgg ccacacacgg 6960
cctggccgca gtgaattccg tgccatggga caccatcgcc aattatgcca agcccttcct 7020
gaacaaggtg gtgagcacaa ccacaaacat cgtgacacgg tgcctgaacc gggtgtgcac 7080
caattacatg ccatatttct ttacactgct gctgcagctg tgcaccttta caaggtccac 7140
caattctcgc atcaaggcct ccatgcccac cacaatcgcc aagaacacag tgaagagcgt 7200
gggcaagttc tgcctggagg cctcctttaa ctacctgaag tcccccaatt tctctaagct 7260
gatcaacatc atcatctggt ttctgctgct gagcgtgtgc ctgggcagcc tgatctattc 7320
cacagccgcc ctgggcgtgc tgatgagcaa cctgggcatg ccttcctact gcaccggcta 7380
tcgggagggc tacctgaata gcaccaacgt gacaatcgcc acctactgta caggctctat 7440
cccatgcagc gtgtgcctgt ccggcctgga ttctctggac acctatcctt ccctggagac 7500
catccagatc acaatctcct ctttcaagtg ggacctgacc gcctttggcc tggtggcaga 7560
gtggttcctg gcctatatcc tgtttacaag attcttttac gtgctgggcc tggccgccat 7620
catgcagctg ttctttagct acttcgccgt gcactttatc tctaatagct ggctgatgtg 7680
gctgatcatc aacctggtgc agatggcccc catctccgcc atggtgagga tgtatatctt 7740
ctttgcctct ttctactacg tgtggaagag ctacgtgcac gtggtggacg gctgcaatag 7800
ctccacctgc atgatgtgct acaagaggaa ccgcgccaca cgcgtggagt gtaccacaat 7860
cgtgaatggc gtgcggagaa gcttctacgt gtatgccaac ggcggcaagg gcttttgcaa 7920
gctgcacaac tggaattgcg tgaactgtga tacattctgt gccggcagca cctttatctc 7980
cgatgaggtg gcaagggacc tgtccctgca gttcaagaga ccaatcaatc ccaccgatca 8040
gtctagctac atcgtggact ccgtgacagt gaagaacggc tctatccacc tgtatttcga 8100
taaggccggc cagaagacat acgagaggca ctccctgtct cactttgtga atctggacaa 8160
cctgcgcgcc aacaatacca agggcagcct gcccatcaac gtgatcgtgt tcgatggcaa 8220
gtccaagtgc gaggagtcct ctgccaagag cgcctccgtg tactatagcc agctgatgtg 8280
ccagcctatc ctgctgctgg accaggccct ggtgtccgat gtgggcgact ctgccgaggt 8340
ggcagtgaag atgtttgatg cctacgtgaa taccttcagc agcaccttca acgtgccaat 8400
ggagaagctg aagaccctgg tggcaacagc agaggcagag ctggccaaga acgtgtccct 8460
ggacaatgtg ctgtctacct tcatcagcgc cgcccgccag ggctttgtgg attctgacgt 8520
ggagacaaag gatgtggtgg agtgcctgaa gctgagccac cagtccgata tcgaggtgac 8580
cggcgacagc tgtaacaatt atatgctgac ctacaataag gtggagaaca tgacaccccg 8640
ggatctgggc gcctgcatcg actgttctgc cagacacatc aatgcccagg tggccaagag 8700
ccacaatatc gccctgatct ggaacgtgaa ggacttcatg tctctgagcg agcagctgag 8760
gaagcagatc cgctccgccg ccaagaagaa caatctgccc ttcaagctga cctgcgccac 8820
cacaaggcag gtggtgaacg tggtcaccac aaagatcgcc ctgaagggcg gcaagatcgt 8880
gaacaattgg ctgaagcagc tgatcaaggt gaccctggtg ttcctgtttg tggccgccat 8940
cttctacctg atcacccccg tgcacgtgat gtctaagcac acagattttt ctagcgagat 9000
catcggctat aaggccatcg acggaggagt gaccagggat atcgccagca ccgacacatg 9060
cttcgccaat aagcacgccg atttcgacac ctggtttagc cagaggggcg gctcctacac 9120
aaacgacaag gcctgtccac tgatcgcagc cgtgatcacc agggaagtgg gattcgtggt 9180
gcctggactg ccaggaacaa tcctgaggac cacaaatggc gacttcctgc actttctgcc 9240
tcgcgtgttt tccgccgtgg gcaacatctg ctatacccca tctaagctga tcgagtacac 9300
cgatttcgcc acatccgcct gcgtgctggc cgcagagtgt accatcttta aggatgcctc 9360
tggcaagccc gtgccttact gttatgacac aaatgtgctg gagggctctg tggcctatga 9420
gagcctgcgg ccagatacca gatacgtgct gatggacggc agcatcatcc agttccccaa 9480
cacatatctg gagggctctg tgcgggtggt gaccacattt gacagcgagt actgccggca 9540
cggcacctgt gagagatctg aggccggcgt gtgcgtgtcc acatctggca ggtgggtgct 9600
gaacaatgat tactatcgca gcctgcctgg cgtgttctgt ggcgtggacg ccgtgaatct 9660
gctgaccaac atgtttacac ctctgatcca gccaatcggc gccctggata tcagcgcctc 9720
catcgtggca ggaggaatcg tggcaatcgt ggtgacatgc ctggcctact atttcatgcg 9780
gttccggagg gccttcggcg agtactctca cgtggtggcc tttaataccc tgctgttcct 9840
gatgagcttc accgtgctgt gcctgacccc cgtgtatagc ttcctgcctg gcgtgtactc 9900
cgtgatctac ctgtatctga ccttctacct gacaaacgac gtgagctttc tggcccacat 9960
ccagtggatg gtcatgttca cccccctggt gcctttttgg atcacaatcg cctatatcat 10020
ctgcatctcc accaagcact tctattggtt cttttctaat tacctgaagc ggagagtggt 10080
gtttaacggc gtgtctttca gcacctttga ggaggccgcc ctgtgcacat tcctgctgaa 10140
caaggagatg tacctgaagc tgcggtccga cgtgctgctg ccactgaccc agtacaatag 10200
atatctggcc ctgtataaca agtacaagta tttctctggc gccatggata ccacaagcta 10260
cagagaggca gcatgctgtc acctggcaaa ggccctgaat gatttttcca actctggcag 10320
cgacgtgctg taccagcccc ctcagacctc tatcacaagc gccgtgctgc agtaactagc 10380
ataacccctt ggggcctcta aacgggtctt gaggggtttt ttgtctaga 10429
<210> 3
<211> 4012
<212> DNA
<213> 人工序列
<400> 3
gctagcgagg gcccggaaac ctggccctgt cttcttgacg agcattccta ggggtctttc 60
ccctctcgcc aaaggaatgc aaggtctgtt gaatgtcgtg aaggaagcag ttcctctgga 120
agcttcttga agacaaacaa cgtctgtagc gaccctttgc aggcagcgga accccccacc 180
tggcgacagg tgcctctgcg gccaaaagcc acgtgtataa gatacacctg caaaggcggc 240
acaaccccag tgccacgttg tgagttggat agttgtggaa agagtcaaat ggctctcctc 300
aagcgtattc aacaaggggc tgaaggatgc ccagaaggta ccccattgta tgggatctga 360
tctggggcct cggtgcacat gctttacatg tgtttagtcg aggttaaaaa aacgtctagg 420
ccccccgaac cacggggacg tggttttcct ttgaaaaaca cgatgataaa tgagcggctt 480
tcggaagatg gcattcccat ccggcaaggt ggagggatgc atggtgcagg tgacatgtgg 540
caccacaacc ctgaatggcc tgtggctgga cgatgtggtg tattgcccta gacacgtgat 600
ctgtaccagc gaggacatgc tgaacccaaa ttacgaggat ctgctgatca ggaagtccaa 660
ccacaatttc ctggtgcagg caggaaacgt gcagctgcgc gtgatcggcc acagcatgca 720
gaattgcgtg ctgaagctga aggtggacac agccaaccca aagaccccca agtacaagtt 780
tgtgaggatc cagcctggcc agacattctc cgtgctggcc tgctataacg gctctcccag 840
cggcgtgtac cagtgtgcca tgcgccctaa ctttaccatc aagggctctt tcctgaatgg 900
cagctgcggc tccgtgggct ttaacatcga ctatgattgc gtgagcttct gttacatgca 960
ccacatggag ctgccaacag gagtgcacgc aggaaccgac ctggagggaa acttctacgg 1020
ccccttcgtg gacaggcaga ccgcacaggc agcaggcaca gatacaacca tcaccgtgaa 1080
cgtgctggcc tggctgtacg ccgccgtgat caacggcgac cggtggtttc tgaatagatt 1140
cacaaccaca ctgaacgatt tcaatctggt ggccatgaag tacaactatg agccactgac 1200
acaggaccac gtggatatcc tgggaccact gagcgcccag accggaatcg ccgtgctgga 1260
catgtgcgcc tccctgaagg agctgctgca gaacggcatg aatggaagga caatcctggg 1320
aagcgccctg ctggaggacg agtttacccc attcgatgtg gtgagacagt gttccggcgt 1380
gacatttcag gccaccaatt tctctctgct gaagcaggca ggcgatgtgg aggagaaccc 1440
tggaccatcc gccgtgaagc gcacaatcaa gggcacccac cactggctgc tgctgacaat 1500
cctgacctct ctgctggtgc tggtgcagtc tacccagtgg agcctgttct ttttcctgta 1560
tgagaatgcc tttctgccct tcgccatggg catcatcgcc atgtccgcct ttgccatgat 1620
gttcgtgaag cacaagcacg cctttctgtg cctgttcctg ctgccatccc tggccaccgt 1680
ggcctacttc aacatggtgt atatgcctgc ctcttgggtc atgaggatca tgacatggct 1740
ggacatggtg gatacctccc tgtctggctt taagctgaag gactgcgtga tgtatgccag 1800
cgccgtggtg ctgctgatcc tgatgacagc aaggaccgtg tacgacgatg gagcaaggag 1860
agtgtggaca ctgatgaatg tgctgaccct ggtgtacaag gtgtactatg gcaacgccct 1920
ggatcaggcc atctccatgt gggccctgat catctctgtg accagcaatt attccggcgt 1980
ggtgaccaca gtgatgtttc tggcccgggg catcgtgttc atgtgcgtgg agtactgtcc 2040
tatctttttc atcacaggca acaccctgca gtgcatcatg ctggtgtact gttttctggg 2100
ctatttctgc acctgttact ttggcctgtt ctgcctgctg aataggtatt ttcgcctgac 2160
actgggcgtg tacgactatc tggtgtctac ccaggagttc agatacatga acagccaggg 2220
cctgctgccc cctaagaact ccatcgatgc cttcaagctg aatatcaagc tgctgggcgt 2280
gggcggcaag ccatgcatca aggtggccac agtgcagtct aagatgagcg acgtgaagtg 2340
taccagcgtg gtgctgctgt ccgtgctgca gcagctgagg gtggagagct cctctaagct 2400
gtgggcccag tgcgtgcagc tgcacaacga catcctgctg gccaaggata ccacagaggc 2460
cttcgagaag atggtgtccc tgctgtctgt gctgctgagc atgcagggcg ccgtggacat 2520
caataagctg tgcgaggaga tgctggataa ccgcgccaca ctgcaggcca tcgcctctga 2580
gtttagctcc ctgccaagct atgcagcctt cgccaccgca caggaggcat acgagcaggc 2640
cgtggccaat ggcgactccg aggtggtgct gaagaagctg aagaagagcc tgaacgtggc 2700
caagtccgag ttcgaccggg atgccgccat gcagagaaag ctggagaaga tggccgacca 2760
ggccatgaca cagatgtata agcaggccag gtctgaggat aagcgcgcca aggtgaccag 2820
cgccatgcag acaatgctgt ttaccatgct gcggaagctg gacaatgatg ccctgaacaa 2880
tatcatcaac aatgccagag acggctgcgt gcccctgaac atcatccctc tgaccacagc 2940
cgccaagctg atggtggtca tccctgacta caacacatat aagaatacct gtgatggcac 3000
cacattcaca tacgcctctg ccctgtggga gatccagcag gtggtggacg ccgatagcaa 3060
gatcgtgcag ctgagcgaga tctccatgga taactcccca aatctggcat ggccactgat 3120
cgtgaccgcc ctgagggcca atagcgccgt gaagctgcag aacaatgagc tgtccccagt 3180
ggccctgagg cagatgtctt gcgcagcagg aaccacacag acagcctgta ccgacgataa 3240
cgccctggcc tactataata ccacaaaggg aggccggttt gtgctggccc tgctgtctga 3300
cctgcaggat ctgaagtggg ccagattccc taagagcgac ggcaccggca caatctacac 3360
cgagctggag ccaccctgcc ggtttgtgac cgatacacct aagggcccaa aggtgaagta 3420
cctgtatttc atcaagggcc tgaacaatct gaacagggga atggtgctgg gatctctggc 3480
cgcaaccgtg cgcctgcagg caggaaacgc cacagaggtg cccgccaatt ccaccgtgct 3540
gtctttttgt gccttcgccg tggacgcagc aaaggcatac aaggattatc tggcctccgg 3600
cggccagcct atcaccaatt gcgtgaagat gctgtgcacc cacacaggaa ccggacaggc 3660
catcacagtg accccagagg ccaacatgga ccaggagtct tttggcggcg ccagctgctg 3720
tctgtattgc cggtgtcaca tcgaccaccc caatcctaag ggcttctgcg atctgaaggg 3780
caagtacgtg cagatcccta ccacatgtgc caatgatcca gtgggcttta ccctgaagaa 3840
cacagtgtgc accgtgtgcg gcatgtggaa gggctacggc tgcagctgtg accagctgag 3900
agagcccatg ctgcagtccg ccgatgccca gtcttttctg aacggcttcg ccgtgtaact 3960
agcataaccc cttggggcct ctaaacgggt cttgaggggt tttttgtcta ga 4012
<210> 4
<211> 8641
<212> DNA
<213> 人工序列
<400> 4
gctagcgagg gcccggaaac ctggccctgt cttcttgacg agcattccta ggggtctttc 60
ccctctcgcc aaaggaatgc aaggtctgtt gaatgtcgtg aaggaagcag ttcctctgga 120
agcttcttga agacaaacaa cgtctgtagc gaccctttgc aggcagcgga accccccacc 180
tggcgacagg tgcctctgcg gccaaaagcc acgtgtataa gatacacctg caaaggcggc 240
acaaccccag tgccacgttg tgagttggat agttgtggaa agagtcaaat ggctctcctc 300
aagcgtattc aacaaggggc tgaaggatgc ccagaaggta ccccattgta tgggatctga 360
tctggggcct cggtgcacat gctttacatg tgtttagtcg aggttaaaaa aacgtctagg 420
ccccccgaac cacggggacg tggttttcct ttgaaaaaca cgatgataaa tgtcagcaga 480
tgcacaatca tttcttaaca gagtgtgcgg agtgtcagca gcaagactta caccttgcgg 540
aacaggaaca tcaacagatg tagtttatag ggccttcgat atctacaacg ataaagtggc 600
aggatttgca aagttcttaa agaccaattg ctgcagattt caagagaagg acgaggatga 660
taaccttatc gattcatact ttgtggtgaa gaggcataca ttcagcaatt accaacacga 720
agaaacaatc tacaaccttc ttaaagattg ccctgcagtg gcaaagcatg acttcttcaa 780
gttcagaatc gatggagata tggtgcctca catctcaaga caaagactta caaagtatac 840
gatggcagat ctcgtttatg cgttgcgcca tttcgacgag ggtaattgtg acaccctgaa 900
ggagatcctg gtcacgtata attgctgcga tgatgattac tttaacaaga aggactggta 960
tgatttcgta gagaatcctg acattcttag agtgtacgca aaccttggag aaagagtgag 1020
acaagcactc ctaaagacag ttcaattctg cgacgcaatg agaaacgcag gaatcgtggg 1080
agtgcttaca cttgataacc aagatcttaa cggaaactgg tatgactttg gcgactttat 1140
acagacaaca cctggatcag gagtgcctgt ggtggattca tattatagcc tgctgatgcc 1200
tatccttaca cttacaagag cacttacagc agaatcacat gtggataccg acttgaccaa 1260
accctatatt aaatgggatc tgctgaaata tgactttaca gaagaacgac ttaaactctt 1320
cgacagatac tttaaatact gggatcaaac ataccaccct aactgcgtga actgccttga 1380
tgatagatgc atccttcact gcgcaaactt taacgtgctg ttctcgaccg tgtttcctcc 1440
tacatcattt ggacctcttg tgagaaagat ctttgtggac ggagtacctt tcgtcgtatc 1500
aacaggatac cactttagag aacttggagt agtgcataat caagatgtga acctacattc 1560
tagccgatta tcatttaaag aacttctggt ttatgccgcg gaccctgcaa tgcacgcagc 1620
aagtggcaat ttattacttg acaaacggac aacctgtttc tcggttgccg cacttacaaa 1680
caatgtagct ttccagaccg taaagccagg gaatttcaac aaagatttct atgacttcgc 1740
cgtatcaaag ggattcttca aggagggatc atcagtggaa cttaaacact tcttcttcgc 1800
ccaggatgga aacgcagcaa tctcagatta cgattactac agatacaacc ttcctacaat 1860
gtgcgatatc agacaacttc tcttcgtagt tgaagtggtg gataaatact ttgattgcta 1920
cgatggagga tgcatcaacg caaaccaagt gatcgtgaac aacttggata aatccgctgg 1980
attcccgttt aataagtggg gtaaagcccg cctttactac gattcaatgt catacgaaga 2040
tcaagatgca ttattcgctt atacaaagag gaatgtgatc cctacaatca cacaaatgaa 2100
ccttaaatac gcaatctcag caaagaatcg agcaagaaca gtggcaggag tgtcaatctg 2160
ctcaacaatg acaaacagac aatttcacca gaagctcctg aaatcaatcg cagcaacaag 2220
aggagcaaca gtggtgatcg gaacatcaaa gttctatgga ggttggcaca acatgctcaa 2280
gaccgtgtat agcgatgttg agaatccgca tctcatggga tgggattacc ctaaatgcga 2340
tagagctatg cccaatatgc tgagaatcat ggcatcactt gtgcttgcaa gaaagcatac 2400
cacatgctgc tcactttcac acagattcta tcgacttgca aacgaatgcg cacaggtcct 2460
ctccgagatg gtgatgtgcg gcgggagctt gtatgtgaaa ccaggtggaa catcatcagg 2520
agatgcaaca acagcatacg caaactcagt gtttaacatc tgccaagcag tgacagctaa 2580
tgtaaacgct ctcttgagca ctgacggaaa caagatagcc gataaatacg tgcgtaatct 2640
gcagcatcga ctttacgaat gcctttacag aaacagagat gtagacacgg actttgtaaa 2700
tgaattctat gcttacctta gaaagcattt ctccatgatg atactgagtg acgatgctgt 2760
tgtatgtttc aactcaacat acgcatcaca aggacttgtg gcatcaatca agaatttcaa 2820
atcagtgctt tactaccaga ataatgtgtt tatgtcagaa gcaaagtgtt ggacagaaac 2880
tgacctcact aagggccctc acgagttctg tagccaacac acaatgcttg tgaaacaagg 2940
agatgactat gtttatctcc cataccctga tccttcaaga atcttgggtg cagggtgttt 3000
cgtggatgat atcgtgaaga ctgacggaac acttatgatc gaaagatttg tgtcacttgc 3060
aatcgatgca taccctctta caaagcatcc gaaccaagaa tacgcagatg tgtttcacct 3120
ttaccttcaa tacatcagaa agttgcatga tgaacttaca ggacacatgc ttgatatgta 3180
ctcagtgatg cttacaaacg ataacacatc aagatactgg gaacctgaat tctatgaggc 3240
aatgtacaca cctcacacag tgcttcaagc agtgggagca tgcgtgcttt gcaactcaca 3300
aacatcactt agatgcggag catgcatcag aagacctttc ctgtgttgca aatgctgcta 3360
cgatcacgtg atctcaacat cacacaaact tgtgctttca gtgaaccctt acgtgtgcaa 3420
cgcaccaggc tgtgacgtaa ctgacgttac gcagctctat cttggaggaa tgtcatacta 3480
ctgcaaatca cacaaacctc ctatctcatt tcctctttgc gcaaacggac aagtgtttgg 3540
actttacaag aatacttgcg tgggatcaga taacgtgaca gatttcaatg ctatcgcaac 3600
atgcgattgg acaaacgcag gagattacat ccttgcaaac acatgcacag agcgtctgaa 3660
gttgtttgcg gccgaaacac ttaaagcaac agaagaaaca tttaaacttt catacggaat 3720
cgcaacagtg agagaggtcc tatcggacag ggaactccac ctttcatggg aagtgggcaa 3780
accacgcccg ccgcttaaca gaaactacgt gtttacagga tacagagtga caaagaattc 3840
taaggtacag atcggagaat acacatttga gaagggcgac tacggagacg ccgtggtgta 3900
cagagggacg actacgtata aacttaacgt gggagattac tttgtgctta catcacacac 3960
agtgatgcct ctttcagcac ctacacttgt gcctcaagag cattatgtcc gaataacggg 4020
tctctatccg acacttaaca tctcagatga attctcgagt aacgtggcaa actaccagaa 4080
agtgggtatg cagaaatact ccaccttaca gggacctcct ggtacaggaa agtctcattt 4140
cgcgataggt ctagctctct attacccttc agcaagaatc gtgtacacag catgctcaca 4200
cgcagcagtg gatgcacttt gcgagaaggc gctgaaatac cttcctatcg ataaatgctc 4260
aagaatcatc cctgcaagag caagagtgga atgctttgat aaatttaaag tgaactcaac 4320
acttgaacaa tacgtgttct gtactgtaaa tgctctgcct gaaactaccg cggatatcgt 4380
ggtgttcgac gagatatcca tggcaacaaa ctacgaccta tcggtcgtaa acgcgcggct 4440
aagagcaaag cattatgtgt acatcggaga tcctgcacaa cttcctgcac ctagaacatt 4500
actaactaaa gggacgctcg aacctgaata ctttaacagt gtttgtcgcc taatgaagac 4560
gatcgggccg gacatgtttc ttggaacatg cagaagatgc cctgcagaaa tcgtggatac 4620
agtgtcagca cttgtgtacg ataacaaact taaagcacac aaagacaagt cggctcagtg 4680
tttcaagatg ttttacaaag gagtgatcac acacgatgtg tcatcagcaa tcaacagacc 4740
tcaaatcgga gtggtgagag aatttcttac aagaaaccct gcatggagaa aggcggtctt 4800
cataagtcct tacaactcac agaatgccgt ggcatcaaag atactcgggc ttcctacaca 4860
aacagtggat tcatcacaag gatcagaata cgattacgtg atctttacac aaacaacaga 4920
aacagcacac tcatgcaacg tgaacagatt taacgtggca atcacaagag caaaggtagg 4980
gatcctctgt atcatgtcag atagagatct ttacgataaa cttcaattta catcacttga 5040
aatccctaga agaaacgtgg cgactctgca ggctgagaac gtgacaggat tgttcaagga 5100
ctgctcaaag gtaattacgg gtttacatcc gacacaagca cctacacacc tttcagtgga 5160
tacaaagttc aagactgaag gactttgcgt ggatatccct ggaatcccta aagatatgac 5220
atacagaaga cttatctcaa tgatgggatt taagatgaat taccaagtga acggataccc 5280
taacatgttt atcacaagag aagaagcaat cagacacgtg agagcatgga taggcttcga 5340
cgtcgaggga tgccacgcaa caagagaagc agtgggaaca aaccttcctc ttcaacttgg 5400
attctccact ggagtgaacc ttgtggcagt gcctacagga tacgtggata cacctaacaa 5460
cacagatttc tcgcgagtgt cagcaaagcc accacctgga gatcaattta aacaccttat 5520
ccctcttatg tacaaaggac ttccttggaa cgtggtgaga atcaagatag tccaaatgct 5580
atccgatacc ttaaagaatc ttagtgaccg tgtcgtattt gtgctttggg cacacggatt 5640
tgaacttaca tcaatgaaat actttgtgaa gatcggtccc gagcgtacat gctgcctttg 5700
cgatagaaga gctacgtgtt tcagtaccgc ttcagataca tacgcatgct ggcaccactc 5760
aataggcttc gattacgttt ataatccgtt catgatagat gtgcaacaat ggggattcac 5820
gggcaatctg cagagcaacc acgatcttta ctgccaagtg cacggaaacg cacacgtggc 5880
atcatgcgat gcaatcatga caagatgcct tgcagtgcac gaatgctttg tgaagcgggt 5940
cgattggaca atcgaatacc ctatcatcgg agatgaactt aagataaatg cagcatgcag 6000
aaaggtccag cacatggtgg tgaaagcagc acttcttgca gataaatttc ctgtgcttca 6060
cgatatcgga aaccctaaag caatcaaatg cgtgcctcaa gcagatgtgg aatggaaatt 6120
ctatgacgca caaccttgct cagataaagc atacaagata gaggaactat tctatagtta 6180
cgcaacacac tcagataaat ttacagatgg agtgtgcctg ttctggaatt gcaacgtgga 6240
tagataccct gcaaactcaa tcgtgtgcag atttgataca agagtgcttt caaaccttaa 6300
ccttccaggt tgtgacggcg gcagtctata tgttaataag cacgcatttc acacacctgc 6360
attcgataag tccgcattcg tcaatttaaa gcagctacct ttcttctatt attcagattc 6420
accttgcgaa tcacacggaa agcaggttgt cagtgacatc gattacgtgc ctcttaaatc 6480
agcaacatgt attaccaggt gtaatcttgg aggagccgtc tgtcgacatc atgcaaacga 6540
atacagactt taccttgatg catacaacat gatgatctcc gccgggttct ccctatgggt 6600
gtacaaacaa tttgatacat acaacctttg gaacacattt acaagacttc aatcacttga 6660
gaacgttgcg ttcaatgtag tcaataaggg acacttcgac ggtcaacagg gtgaggttcc 6720
tgtgtcaatc atcaacaata ccgtttatac taaagttgac ggcgtggatg tggaactctt 6780
cgagaataag actacgcttc ctgtgaatgt tgccttcgag ttgtgggcaa agcgcaatat 6840
caaacctgtg cctgaagtga agatactcaa taaccttgga gtggatatcg cagcaaacac 6900
agtgatctgg gattacaaga gggacgcacc tgcacacatc tcaacaatcg gagtgtgctc 6960
aatgacagat atcgcaaaga agccgactga aacaatctgc gcacctctta ctgtattctt 7020
cgacggaaga gtggatggac aagtggattt attccgaaat gcaagaaacg gagtgcttat 7080
cacagaagga tcagtgaaag gacttcaacc ttcagtggga cctaaacaag catcacttaa 7140
cggagtgact ctgataggcg aggccgtgaa gactcagttt aactactaca agaaagtaga 7200
cggtgtcgtc cagcagctgc ccgagaccta tttcacacaa tcacggaatc tgcaggagtt 7260
caaacctaga tcacaaatgg aaatcgattt cctggagctt gcaatggatg aatttatcga 7320
aagatacaaa cttgaaggat acgcatttga acacatcgtg tacggagatt tcagtcattc 7380
acaacttgga ggacttcacc ttcttattgg cctagccaaa cgtttcaaag aatcaccttt 7440
cgagctcgaa gatttcattc caatggattc aacagtgaag aattatttca ttactgacgc 7500
ccagacggga tcatcaaagt gtgtatgctc agtgatcgat ctactactag acgatttcgt 7560
tgaaattatt aaatcacaag acttgagtgt agttagtaag gttgtgaagg tcacaatcga 7620
ttacacagaa atctcattta tgctttggtg caaagatgga cacgtggaaa cattctatcc 7680
caaacttcaa tcatcacaag catggcaacc tggagtggcc atgccgaatt tgtataagat 7740
gcagagaatg cttcttgaga agtgtgacct tcagaattat ggagattcag caacacttcc 7800
taaaggaatc atgatgaacg tggcaaagta tactcaactt tgccaatacc ttaacacact 7860
tacacttgca gtgccttaca acatgagagt gatccacttc ggtgcagggt cggacaaagg 7920
agtggcacct ggtactgctg tccttagaca atggcttcct acaggaacac ttcttgtgga 7980
ttcagatctt aacgatttcg tctccgatgc agattcaacc ctcattggtg actgtgcaac 8040
agtgcacaca gcaaacaagt gggacttaat aatatcagat atgtacgatc ctaagactaa 8100
gaatgtaacg aaagagaatg actcaaagga aggtttcttc acctatatct gcggatttat 8160
ccaacagaag ttagctcttg gaggatcagt ggcaatcaag attacggaac actcatggaa 8220
cgcagatctt tacaaactta tgggacactt tgcatggtgg accgcgttcg ttacaaacgt 8280
aaacgcgtcg tcctcagaag catttcttat cggatgcaac taccttggga aaccaagaga 8340
gcagatcgat ggatacgtga tgcacgcaaa ctacatcttc tggaggaaca caaaccctat 8400
ccaactttca tcatactcac tcttcgacat gtcaaagttc ccgcttaaac ttagagggac 8460
tgccgtaatg tcgcttaaag aaggacaaat caacgatatg atactcagcc tcctaagtaa 8520
agggaggctt atcatcagag agaataatag agtggtgatc tcatcagatg tgcttgtgaa 8580
caactaacta gcataacccc ttggggcctc taaacgggtc ttgaggggtt ttttgtctag 8640
a 8641
Claims (1)
1.阿霉素或其药学上可接受的盐作为唯一活性成分在制备治疗SARS-CoV-2感染药物中的应用，所述药物通过抑制SARS-CoV-2 RNA复制达到治疗目的。
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| CN114377010B (zh) * | 2022-01-26 | 2023-04-07 | 中山大学 | Pkc激酶抑制剂在制备具有防治新型冠状病毒感染相关疾病的药物中的应用 |
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| CN101428025A (zh) * | 2008-10-24 | 2009-05-13 | 中国人民解放军第四军医大学 | 抗疟药物羟基氯喹的一种新用途 |
| WO2010048603A1 (en) * | 2008-10-24 | 2010-04-29 | Indiana University Research & Technology Corporation | Materials and methods for sensitizing multidrug resistant cells |
| CN105287383A (zh) * | 2015-11-19 | 2016-02-03 | 吉林大学 | 新型米托蒽醌脂质体联合化疗药物在抗肿瘤治疗中的应用 |
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| CN107184586B (zh) * | 2017-06-25 | 2020-06-19 | 浙江大学 | 血红素加氧酶-1抑制剂在制备抑制阿霉素心脏毒性药物中的应用 |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN101428025A (zh) * | 2008-10-24 | 2009-05-13 | 中国人民解放军第四军医大学 | 抗疟药物羟基氯喹的一种新用途 |
| WO2010048603A1 (en) * | 2008-10-24 | 2010-04-29 | Indiana University Research & Technology Corporation | Materials and methods for sensitizing multidrug resistant cells |
| CN105287383A (zh) * | 2015-11-19 | 2016-02-03 | 吉林大学 | 新型米托蒽醌脂质体联合化疗药物在抗肿瘤治疗中的应用 |
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| Title |
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| DNA拓扑异构酶Ⅱ抑制剂研究进展;王汝冰等;《海峡药学》;20090415(第04期);全文 * |
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