CN114573600B - 四个具有激活潜伏hiv作用的二萜类成分的制备方法和应用 - Google Patents
四个具有激活潜伏hiv作用的二萜类成分的制备方法和应用 Download PDFInfo
- Publication number
- CN114573600B CN114573600B CN202210239249.2A CN202210239249A CN114573600B CN 114573600 B CN114573600 B CN 114573600B CN 202210239249 A CN202210239249 A CN 202210239249A CN 114573600 B CN114573600 B CN 114573600B
- Authority
- CN
- China
- Prior art keywords
- diterpenoid
- components
- component
- latent hiv
- chamaedaphnelide
- 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
- 230000003213 activating effect Effects 0.000 title claims abstract description 20
- 150000004141 diterpene derivatives Chemical class 0.000 title claims abstract description 16
- 238000002360 preparation method Methods 0.000 title claims abstract description 16
- -1 diterpenoid compounds Chemical class 0.000 claims abstract description 13
- 239000003814 drug Substances 0.000 claims abstract description 12
- 230000000694 effects Effects 0.000 claims abstract description 8
- 239000008194 pharmaceutical composition Substances 0.000 claims abstract description 7
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 claims description 27
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 claims description 21
- YMWUJEATGCHHMB-UHFFFAOYSA-N Dichloromethane Chemical compound ClCCl YMWUJEATGCHHMB-UHFFFAOYSA-N 0.000 claims description 15
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 claims description 14
- 239000000546 pharmaceutical excipient Substances 0.000 claims description 12
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 11
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 11
- 238000010898 silica gel chromatography Methods 0.000 claims description 9
- 238000010828 elution Methods 0.000 claims description 8
- 239000003208 petroleum Substances 0.000 claims description 8
- 238000002953 preparative HPLC Methods 0.000 claims description 8
- 239000000284 extract Substances 0.000 claims description 7
- 238000000926 separation method Methods 0.000 claims description 7
- 238000004440 column chromatography Methods 0.000 claims description 6
- XELZGAJCZANUQH-UHFFFAOYSA-N methyl 1-acetylthieno[3,2-c]pyrazole-5-carboxylate Chemical compound CC(=O)N1N=CC2=C1C=C(C(=O)OC)S2 XELZGAJCZANUQH-UHFFFAOYSA-N 0.000 claims description 6
- 239000003937 drug carrier Substances 0.000 claims description 3
- 238000002156 mixing Methods 0.000 claims description 3
- 239000002904 solvent Substances 0.000 claims description 3
- 241001465754 Metazoa Species 0.000 claims description 2
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 claims description 2
- 239000002024 ethyl acetate extract Substances 0.000 claims description 2
- 238000001914 filtration Methods 0.000 claims description 2
- 238000004128 high performance liquid chromatography Methods 0.000 claims description 2
- 238000000034 method Methods 0.000 claims description 2
- 231100000252 nontoxic Toxicity 0.000 claims description 2
- 230000003000 nontoxic effect Effects 0.000 claims description 2
- 238000010992 reflux Methods 0.000 claims description 2
- 150000003839 salts Chemical class 0.000 claims description 2
- 239000000741 silica gel Substances 0.000 claims description 2
- 229910002027 silica gel Inorganic materials 0.000 claims description 2
- 229940126214 compound 3 Drugs 0.000 claims 1
- 230000008030 elimination Effects 0.000 claims 1
- 238000003379 elimination reaction Methods 0.000 claims 1
- 150000001875 compounds Chemical class 0.000 abstract description 30
- 229940079593 drug Drugs 0.000 abstract description 8
- 241001263989 Wikstroemia Species 0.000 abstract description 5
- 239000003795 chemical substances by application Substances 0.000 abstract description 3
- 238000007142 ring opening reaction Methods 0.000 abstract description 2
- 240000003538 Chamaemelum nobile Species 0.000 abstract 1
- 235000007866 Chamaemelum nobile Nutrition 0.000 abstract 1
- 235000007232 Matricaria chamomilla Nutrition 0.000 abstract 1
- 241000725303 Human immunodeficiency virus Species 0.000 description 20
- 210000004027 cell Anatomy 0.000 description 8
- MNXGCOMZNRZZJF-UHFFFAOYSA-N 6alpha,7alpha-epoxy-5beta-hydroxy-12-deoxyphorbol-13-decanoate Natural products C1C(C)C2(O)C3C=C(C)C(=O)C3(O)C(O)C3(CO)OC3C2C2C1(OC(=O)CCCCCCCCC)C2(C)C MNXGCOMZNRZZJF-UHFFFAOYSA-N 0.000 description 6
- 208000030507 AIDS Diseases 0.000 description 6
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 description 6
- 230000006870 function Effects 0.000 description 6
- 239000000843 powder Substances 0.000 description 6
- 230000003612 virological effect Effects 0.000 description 5
- 241000700605 Viruses Species 0.000 description 4
- ROAYSRAUMPWBQX-UHFFFAOYSA-N ethanol;sulfuric acid Chemical compound CCO.OS(O)(=O)=O ROAYSRAUMPWBQX-UHFFFAOYSA-N 0.000 description 4
- 239000008187 granular material Substances 0.000 description 4
- 238000002347 injection Methods 0.000 description 4
- 239000007924 injection Substances 0.000 description 4
- 239000007788 liquid Substances 0.000 description 4
- 238000004020 luminiscence type Methods 0.000 description 4
- 210000001744 T-lymphocyte Anatomy 0.000 description 3
- 230000004913 activation Effects 0.000 description 3
- 238000011225 antiretroviral therapy Methods 0.000 description 3
- 239000002775 capsule Substances 0.000 description 3
- 239000013592 cell lysate Substances 0.000 description 3
- 238000002114 high-resolution electrospray ionisation mass spectrometry Methods 0.000 description 3
- 239000007787 solid Substances 0.000 description 3
- 239000000243 solution Substances 0.000 description 3
- 241000713772 Human immunodeficiency virus 1 Species 0.000 description 2
- 210000000987 immune system Anatomy 0.000 description 2
- OIPPWFOQEKKFEE-UHFFFAOYSA-N orcinol Chemical compound CC1=CC(O)=CC(O)=C1 OIPPWFOQEKKFEE-UHFFFAOYSA-N 0.000 description 2
- 230000000284 resting effect Effects 0.000 description 2
- 230000001225 therapeutic effect Effects 0.000 description 2
- 241000723435 Chamaedaphne Species 0.000 description 1
- 108010077544 Chromatin Proteins 0.000 description 1
- 208000035473 Communicable disease Diseases 0.000 description 1
- IGXWBGJHJZYPQS-SSDOTTSWSA-N D-Luciferin Chemical compound OC(=O)[C@H]1CSC(C=2SC3=CC=C(O)C=C3N=2)=N1 IGXWBGJHJZYPQS-SSDOTTSWSA-N 0.000 description 1
- CYCGRDQQIOGCKX-UHFFFAOYSA-N Dehydro-luciferin Natural products OC(=O)C1=CSC(C=2SC3=CC(O)=CC=C3N=2)=N1 CYCGRDQQIOGCKX-UHFFFAOYSA-N 0.000 description 1
- BJGNCJDXODQBOB-UHFFFAOYSA-N Fivefly Luciferin Natural products OC(=O)C1CSC(C=2SC3=CC(O)=CC=C3N=2)=N1 BJGNCJDXODQBOB-UHFFFAOYSA-N 0.000 description 1
- 208000031886 HIV Infections Diseases 0.000 description 1
- 208000037357 HIV infectious disease Diseases 0.000 description 1
- 241000700721 Hepatitis B virus Species 0.000 description 1
- 208000032420 Latent Infection Diseases 0.000 description 1
- DDWFXDSYGUXRAY-UHFFFAOYSA-N Luciferin Natural products CCc1c(C)c(CC2NC(=O)C(=C2C=C)C)[nH]c1Cc3[nH]c4C(=C5/NC(CC(=O)O)C(C)C5CC(=O)O)CC(=O)c4c3C DDWFXDSYGUXRAY-UHFFFAOYSA-N 0.000 description 1
- 102000006833 Multifunctional Enzymes Human genes 0.000 description 1
- 108010047290 Multifunctional Enzymes Proteins 0.000 description 1
- 108010047956 Nucleosomes Proteins 0.000 description 1
- 108091023040 Transcription factor Proteins 0.000 description 1
- 102000040945 Transcription factor Human genes 0.000 description 1
- 230000021736 acetylation Effects 0.000 description 1
- 238000006640 acetylation reaction Methods 0.000 description 1
- 230000009471 action Effects 0.000 description 1
- 239000012190 activator Substances 0.000 description 1
- 239000002671 adjuvant Substances 0.000 description 1
- 239000000443 aerosol Substances 0.000 description 1
- 230000000798 anti-retroviral effect Effects 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 238000007664 blowing Methods 0.000 description 1
- 239000007853 buffer solution Substances 0.000 description 1
- 230000010307 cell transformation Effects 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 238000012512 characterization method Methods 0.000 description 1
- 210000003483 chromatin Anatomy 0.000 description 1
- 229940125904 compound 1 Drugs 0.000 description 1
- 229940125782 compound 2 Drugs 0.000 description 1
- 238000012258 culturing Methods 0.000 description 1
- 230000034994 death Effects 0.000 description 1
- 231100000517 death Toxicity 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
- 239000003085 diluting agent Substances 0.000 description 1
- 208000037771 disease arising from reactivation of latent virus Diseases 0.000 description 1
- 238000004090 dissolution Methods 0.000 description 1
- 239000012153 distilled water Substances 0.000 description 1
- 239000002552 dosage form Substances 0.000 description 1
- 238000002651 drug therapy Methods 0.000 description 1
- 238000001035 drying Methods 0.000 description 1
- 239000000839 emulsion Substances 0.000 description 1
- 230000004049 epigenetic modification Effects 0.000 description 1
- 239000000469 ethanolic extract Substances 0.000 description 1
- SRCZQMGIVIYBBJ-UHFFFAOYSA-N ethoxyethane;ethyl acetate Chemical compound CCOCC.CCOC(C)=O SRCZQMGIVIYBBJ-UHFFFAOYSA-N 0.000 description 1
- 239000000945 filler Substances 0.000 description 1
- 230000008014 freezing Effects 0.000 description 1
- 238000007710 freezing Methods 0.000 description 1
- 238000000227 grinding Methods 0.000 description 1
- 239000001963 growth medium Substances 0.000 description 1
- 231100000206 health hazard Toxicity 0.000 description 1
- 208000033519 human immunodeficiency virus infectious disease Diseases 0.000 description 1
- 230000036737 immune function Effects 0.000 description 1
- 238000001727 in vivo Methods 0.000 description 1
- 208000015181 infectious disease Diseases 0.000 description 1
- 230000002401 inhibitory effect Effects 0.000 description 1
- 230000010354 integration Effects 0.000 description 1
- 239000007927 intramuscular injection Substances 0.000 description 1
- 238000010255 intramuscular injection Methods 0.000 description 1
- 239000007928 intraperitoneal injection Substances 0.000 description 1
- 238000001990 intravenous administration Methods 0.000 description 1
- 230000007246 mechanism Effects 0.000 description 1
- 238000011034 membrane dialysis Methods 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 230000009456 molecular mechanism Effects 0.000 description 1
- 210000004400 mucous membrane Anatomy 0.000 description 1
- 101150015886 nuc-1 gene Proteins 0.000 description 1
- 210000001623 nucleosome Anatomy 0.000 description 1
- 230000003389 potentiating effect Effects 0.000 description 1
- 238000004382 potting Methods 0.000 description 1
- 238000002203 pretreatment Methods 0.000 description 1
- 230000001566 pro-viral effect Effects 0.000 description 1
- 230000010076 replication Effects 0.000 description 1
- 238000007873 sieving Methods 0.000 description 1
- 239000007921 spray Substances 0.000 description 1
- 239000008227 sterile water for injection Substances 0.000 description 1
- 230000001954 sterilising effect Effects 0.000 description 1
- 238000004659 sterilization and disinfection Methods 0.000 description 1
- 239000007929 subcutaneous injection Substances 0.000 description 1
- 238000010254 subcutaneous injection Methods 0.000 description 1
- 239000000758 substrate Substances 0.000 description 1
- 239000000725 suspension Substances 0.000 description 1
- 239000006188 syrup Substances 0.000 description 1
- 235000020357 syrup Nutrition 0.000 description 1
- 239000003826 tablet Substances 0.000 description 1
- 238000010257 thawing Methods 0.000 description 1
- 238000011285 therapeutic regimen Methods 0.000 description 1
- 231100000331 toxic Toxicity 0.000 description 1
- 230000002588 toxic effect Effects 0.000 description 1
- 108091006106 transcriptional activators Proteins 0.000 description 1
- 230000002103 transcriptional effect Effects 0.000 description 1
- 230000029812 viral genome replication Effects 0.000 description 1
- 238000005406 washing Methods 0.000 description 1
- 239000008215 water for injection Substances 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D493/00—Heterocyclic compounds containing oxygen atoms as the only ring hetero atoms in the condensed system
- C07D493/22—Heterocyclic compounds containing oxygen atoms as the only ring hetero atoms in the condensed system in which the condensed system contains four or more hetero rings
-
- 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
- A61P31/12—Antivirals
- A61P31/14—Antivirals for RNA viruses
- A61P31/18—Antivirals for RNA viruses for HIV
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
- C07C67/00—Preparation of carboxylic acid esters
- C07C67/48—Separation; Purification; Stabilisation; Use of additives
- C07C67/56—Separation; Purification; Stabilisation; Use of additives by solid-liquid treatment; by chemisorption
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
- C07C69/00—Esters of carboxylic acids; Esters of carbonic or haloformic acids
- C07C69/02—Esters of acyclic saturated monocarboxylic acids having the carboxyl group bound to an acyclic carbon atom or to hydrogen
- C07C69/22—Esters of acyclic saturated monocarboxylic acids having the carboxyl group bound to an acyclic carbon atom or to hydrogen having three or more carbon atoms in the acid moiety
- C07C69/33—Esters of acyclic saturated monocarboxylic acids having the carboxyl group bound to an acyclic carbon atom or to hydrogen having three or more carbon atoms in the acid moiety esterified with hydroxy compounds having more than three hydroxy groups
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D301/00—Preparation of oxiranes
- C07D301/32—Separation; Purification
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D303/00—Compounds containing three-membered rings having one oxygen atom as the only ring hetero atom
- C07D303/02—Compounds containing oxirane rings
- C07D303/04—Compounds containing oxirane rings containing only hydrogen and carbon atoms in addition to the ring oxygen atoms
- C07D303/06—Compounds containing oxirane rings containing only hydrogen and carbon atoms in addition to the ring oxygen atoms in which the oxirane rings are condensed with a carbocyclic ring system having three or more relevant rings
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07B—GENERAL METHODS OF ORGANIC CHEMISTRY; APPARATUS THEREFOR
- C07B2200/00—Indexing scheme relating to specific properties of organic compounds
- C07B2200/07—Optical isomers
-
- 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
Abstract
本发明属于药物技术领域,具体涉及四个具有激活潜伏HIV作用的二萜类成分的制备方法和应用。为解决现有技术中激活剂无效或激活剂有效但副作用大的问题,本发明从瑞香科荛花属植物河朔荛花(Wikstroemia chamaedaphneMeisn.)枝叶中分离得到2个新的A环开环的瑞香烷型二萜化合物chamaedaphnelideA(1)和epi‑chamaedaphnelide A(2),以及1个新的惕各烷型二萜化合物chamaedaphnelide B(3),该类化合物具有高效的激活潜伏HIV病毒的作用,因此将类化合物及含有该类化合物的药物组合物应用在制备清除潜伏HIV病毒药物中。
Description
技术领域
本发明属于药物技术领域,具体涉及四个具有激活潜伏HIV作用的二萜类成分的制备方法和应用。
背景技术
获得性免疫缺陷综合征(AIDS)是由HIV感染引起的一种严重危害人们生命健康的传染性疾病。据WHO统计全球艾滋病患者已超过4000万,每年新增患者500万,而每年死亡约300万。
目前,艾滋病临床治疗方法主要是高效抗逆转录病毒疗法(Highly activeantiretroviral therapy,HAART),该疗法不仅有效控制HIV复制,并且能重建AIDS 患者的免疫功能,为AIDS的治疗打开了希望之门。
人们曾寄希望于凭借HAART完全清除体内的HIV从而达到彻底治愈AIDS 的目的。但随后的实践证明,虽然HAART可以最大限度地抑制患者体内病毒复制,使血浆病毒载量(virus load)降低至现有常规检测方法测不出的水平,但感染者体内仍有病毒持续存在,一旦停止药物治疗,病毒载量又会反弹到治疗前水平。
HIV难以在体内被完全清除的一个重要原因是HIV-1能够潜伏在静息的记忆CD4+T细胞中,该潜伏感染细胞是由一小部分HIV感染的活化CD4+T细胞转化而产生的,其整合的前病毒缺乏转录活性,因此不会被免疫系统和抗逆转录酶病毒的药物攻击。尽管感染个体携带潜伏感染细胞数量较少,但衰减率是如此之慢,以至于欲在个体生存期内仅靠HAART治疗将其彻底清除是不可能的。因此,HIV潜伏感染的静息CD4+T细胞是构成机体内病毒储藏库(reservoir) 的主要部分,同时也是目前临床治疗不能彻底清除HIV的巨大障碍。
一般认为,HIV-1潜伏感染细胞形成的分子机制与整合位点处的染色质状态、抑制性核小体nuc-1的存在,以乙酰化为代表的表观遗传修饰、宿主转录因子如NF-KB以及病毒转录活化因子Tat等因素有关。据此机制,研究者提出了清除潜伏病毒储藏库的治疗策略,即试图通过药物诱导HIV潜伏感染细胞的前病毒表达,使其潜伏病毒再次激活,同时结合高效抗逆转录病毒疗法及在人体免疫系统作用下,来杀死激活的潜伏感染的细胞,以此加速病毒储藏库的清除。尽管该策略在临床上已有几个治疗方案,但其结果仍未尽人意,不是激活剂无效,就是虽有效但其毒副作用大。因而,研发安全有效的能消除病毒储藏库的试剂及相应的治疗方案已是当务之急。
本发明从瑞香科荛花属植物河朔荛花(Wikstroemia chamaedaphne Meisn.) 枝叶中分离得到2个新的A环开环的瑞香烷型二萜化合物chamaedaphnelide A(1)和epi-chamaedaphnelide A(2),以及1个新的惕各烷型二萜化合物 chamaedaphnelide B(3),该类化合物具有高效的激活潜伏HIV病毒的作用,该化合物至今尚未见到相关报道。
发明内容
针对上述问题本发明提供了四个具有激活潜伏HIV作用的二萜类成分的制备方法和应用。
为了达到上述目的,本发明采用了下列技术方案:
四个具有激活潜伏HIV作用的二萜类成分,所述二萜类成分为chamaedaphnelideA(1)、epi-chamaedaphnelideA(2)、chamaedaphnelide B(3)、和6α,7α-epoxy-5β-hydroxy 12-deoxyphorbol-13-decanoate(4),其结构式为:
四个具有激活潜伏HIV作用的二萜类成分的制备方法,包括以下步骤:
(1)取河朔荛花枝叶,用10倍重量的95%乙醇回流提取(体积分数),滤过,合并提取液,减压浓缩,将所得浸膏加入3-5倍重量水,依次用石油醚,乙酸乙酯萃取,萃取液回收溶剂,减压浓缩得各萃取物;
(2)萃取物经正相硅胶柱色谱梯度洗脱,得到6个组分A1~A6,A4组分经硅胶柱色谱分离洗脱,得到A41~A45五个组分,A43经Sephadex LH-20柱色谱分离,继续得到3个组分A431~A433,A432组分经半制备高效液相色谱分离得到二萜类化合物1和二萜类化合物2;A5组分经Sephadex LH-20柱色谱分离,继续得到5个组分A51~A55,A53组分经半制备高效液相色谱分离得到化合物二萜类化合物3和二萜类化合物4。
进一步,所述正相硅胶柱色谱梯度洗脱采用石油醚和乙酸乙酯进行梯度洗脱,石油醚和乙酸乙酯的体积比为1:1~2;
所述A4组分经硅胶柱色谱分离洗脱采用二氯甲烷和甲醇洗脱,二氯甲烷和甲醇的体积比为8:1~1:3;
所述A432组分经半制备高效液相色谱分离采用55%甲醇-45%水(体积分数);
所述A53组分经半制备高效液相色谱分离采用55%甲醇-45%水(体积分数)。
四个具有激活潜伏HIV作用的二萜类成分的应用,在制备清除潜伏HIV病毒药物中的应用。
进一步,所述药物以二萜化合物直接使用,或以药学上可接受的盐以及药物组合物的形式使用。
进一步,所述药物组合物含有0.1~99%的二萜类化物(优选为0.5~90%,质量分数),其余为药物学上可接受的、对人和动物无毒和惰性的可药用载体和/ 或赋形剂
所述的药用载体或赋形剂是一种或多种固体、半固体和液体稀释剂、填料以及药物制品辅剂。将本发明的药物组合物以单位体重服用量的形式使用。苔黑酚-1-氧-β-D-葡萄糖吡喃苷、苔黑酚及其衍生物的组合物采用制药和食品领域公认的方法制备成各种剂型、如液体制剂(注射剂、混悬剂、乳剂、溶液剂、糖浆剂等)、固体制剂(片剂、胶囊剂、颗粒剂,冲剂等)、喷剂、气雾剂等。本发明的药物可经注射(静脉注射、静脉滴注、肌肉注射、腹腔注射、皮下注射)和口服、舌下给药、粘膜透析等给药途径进行抗乙肝病毒的治疗。
与现有技术相比本发明具有以下优点:
本发明所制备的化合物1-3为新化合物,即目前未有这类结构报道,包括其制备方法和激活潜伏HIV活性都是新颖的。化合物4则是已知化合物,之前有被报道过该结构,但是未报道过化合物4可以激活潜伏HIV作用。
附图说明
图1为化合物1-4在Hela-NH2和J-Lat A2激活潜伏HIV效果(浓度为10 μM);
图2为化合物1-4在J-LatA2激活潜伏HIV的荧光共聚焦图。
具体实施方式
实施例1
chamaedaphnelideA(1)、epi-chamaedaphnelideA(2)、chamaedaphnelide B (3)、和6α,7α-epoxy-5β-hydroxy 12-deoxyphorbol-13-decanoate(4)对激活潜伏 HIV的效果。
(1)接种HeLa-NH2或者J-LatA2细胞于96孔培养板中,培养12h后,加入不同的药物处理,设置三组平行,空白组做相同处理;
(2)收细胞,弃去培养基,用PBS缓冲液洗3次,吸净PBS;
(3)加入100μL细胞裂解液,-80℃反复冻融,溶解后吹打混匀细胞,不要产生气泡;
(4)实验组从步骤3细胞裂解液中吸取50μL转移至荧光白板中,对照组加入50μL细胞裂解液。所有孔中加入适量萤光素底物,检测发光强度在多功能酶标仪上进行;
(5)用实验组测定得到的发光强度除以对照组的读数,得到发光强度的变化倍数,即fold ofchange(fold ofchange一不同加药组发光信号强度/空白组发光信号强度),以指示药物对潜伏HIV的激活强度。
实验结果如图1和图2所示(****p<0.0001),4个化合物在两个浓度下均显示出激活效果,其中chamaedaphnelide A(1)、chamaedaphnelide B(3)和 6α,7α-epoxy-5β-hydroxy 12-deoxyphorbol-13-decanoate(4)显示出高效的激活效果。
实施例2
化合物chamaedaphnelide A(1)、epi-chamaedaphnelide A(2)、chamaedaphnelide B(3)、和6α,7α-epoxy-5β-hydroxy 12-deoxyphorbol-13-decanoate(4)的制备及表征
取河朔荛花枝叶500g,用95%乙醇(5L*2)回流提取两次,每次3小时,合并乙醇提液,减压回收至无醇,用加入相同体积的蒸馏水(400mL)溶解,然后依次用石油醚(1L)和乙酸乙酯(1L)各萃取3次,将乙酸乙酯萃取液浓缩干燥成浸膏(80g),浸膏用80%乙醇溶解吸附于硅胶上,室温放置挥干溶剂,研碎过筛后经硅胶柱层析,用1:1、1:1.5、1:2的石油醚-乙酸乙酯梯度洗脱,得到6 个组分A1-A6,A4组分经硅胶柱色谱分离,采用二氯甲烷:甲醇(8:1~1:3)洗脱得到A41-A45五个组分。A43经Sephadex LH-20柱色谱分离,继续得到3个组分A431-A433。A432组分经半制备高效液相色谱分离(45%甲醇-55%水)得到化合物chamaedaphnelideA(1)和epi-chamaedaphnelideA(2)。A5组分经 Sephadex LH-20柱色谱分离,继续得到5个组分A51-A55。A53组分经半制备高效液相色谱分离(55%甲醇-45%水)得到化合物chamaedaphnelide B(3)和 6α,7α-epoxy-5β-hydroxy 12-deoxyphorbol-13-decanoate(4)。
化合物chamaedaphnelideA:无定形粉末,5%浓硫酸-乙醇显棕色,-22.0 (c0.10,MeOH),IR(KBr)νmax:2929,2858,1709,1460,1388,1363,1223,1100, 1051,912cm-1,HR-ESI-MS m/z 571.2879[M+Na]+,(计算值为571.2883);1H和13C NMR数据,见下表1。
化合物epi-chamaedaphnelide A:无定形粉末,5%浓硫酸-乙醇显棕色,-9.0(c 0.10,MeOH),IR(KBr)νmax:2929,2858,1711,1450,1386,1363,1222, 1079,995cm-1,HR-ESI-MS m/z 571.2879[M+Na]+,(计算值为571.2883);1H 和13C NMR数据,见下表1。
化合物chamaedaphnelide B:色无定形粉末,5%浓硫酸-乙醇显棕色,IR(KBr)νmax:3413,2885,2856,1710,1630,1460,1380,1034,929cm-1;HR-ESI-MS m/z 553.3367[M+H]+(计算值为553.3377),1H和13C NMR数据,见下表1。
化合物6α,7α-epoxy-5β-hydroxy 12-deoxyphorbol-13-decanoate:无定形粉末,5%浓硫酸-乙醇显棕色,该化合物的1H和13C NMR数据与已知化合物 6α,7α-epoxy-5β-hydroxy 12-deoxyphorbol-13-decanoate相一致,故鉴定为 6α,7α-epoxy-5β-hydroxy12-deoxyphorbol-13-decanoate。
表1,化合物1-3的NMR数据
实施例3
按实施例2的方法先制得化合物chamaedaphnelide A(1)、epi-chamaedaphnelideA(2)、chamaedaphnelide B(3)、和6α,7α-epoxy-5β-hydroxy 12-deoxyphorbol-13-decanoate(4),用少量的DMSO溶解后,按常规加注射用水,精滤,灌封灭菌制成注射液。
实施例4
按实施例2的方法先制得化合物chamaedaphnelide A(1)、 epi-chamaedaphnelideA(2)、chamaedaphnelide B(3)、和6α,7α-epoxy-5β-hydroxy 12-deoxyphorbol-13-decanoate(4),用少量的DMSO溶解后,将其溶于无菌注射用水中,搅拌使溶解,用无菌抽滤漏斗过滤,再无菌精滤,分装于安瓿中,低温冷冻干燥后无菌熔封得粉针剂。
实施例5
将实施例2所分离得到的化合物chamaedaphnelide A(1)、 epi-chamaedaphnelideA(2)、chamaedaphnelide B(3)、和6α,7α-epoxy-5β-hydroxy 12-deoxyphorbol-13-decanoate(4),分别按其与赋形剂重量比为9:1的比例加入赋形剂,制成粉剂。
实施例6
按实施例2的方法先制得化合物chamaedaphnelide A(1)、epi-chamaedaphnelideA(2)、chamaedaphnelide B(3)、和6α,7α-epoxy-5β-hydroxy 12-deoxyphorbol-13-decanoate(4),分别按其与赋形剂重量比为5:1的比例加入赋形剂,制粒压片。
实施例7
按实施例2的方法先制得化合物chamaedaphnelide A(1)、 epi-chamaedaphnelideA(2)、chamaedaphnelide B(3)、和6α,7α-epoxy-5β-hydroxy 12-deoxyphorbol-13-decanoate(4),分别按常规口服液制法制成口服液。
实施例8
按实施例2的方法先制得化合物chamaedaphnelide A(1)、 epi-chamaedaphnelideA(2)、chamaedaphnelide B(3)、和6α,7α-epoxy-5β-hydroxy 12-deoxyphorbol-13-decanoate(4),按其与赋形剂重量比为5:1的比例加入赋形剂,制成胶囊。
实施例9
按实施例2的方法先制得化合物chamaedaphnelide A(1)、 epi-chamaedaphnelideA(2)、chamaedaphnelide B(3)、和6α,7α-epoxy-5β-hydroxy 12-deoxyphorbol-13-decanoate(4),按其与赋形剂重量比为3:1的比例加入赋形剂,制成胶囊。
实施例10
按实施例2的方法先制得化合物chamaedaphnelide A(1)、 epi-chamaedaphnelideA(2)、chamaedaphnelide B(3)、和6α,7α-epoxy-5β-hydroxy 12-deoxyphorbol-13-decanoate(4),分别按其与赋形剂重量比为5:1的比例加入赋形剂,制成颗粒剂。
Claims (5)
1.三个具有激活潜伏HIV作用的二萜类成分,其特征在于,所述二萜类成分为chamaedaphnelide A (1)、epi-chamaedaphnelide A(2)、chamaedaphnelide B(3),其结构式为:
。
2.一种权利要求1所述三个具有激活潜伏HIV作用的二萜类成分的制备方法,其特征在于,包括以下步骤:
(1)取河朔荛花枝叶,用10倍重量的95%乙醇回流提取,滤过,合并提取液,减压浓缩,将所得浸膏加入3-5倍重量水,依次用石油醚,乙酸乙酯萃取,萃取液回收溶剂,减压浓缩得各萃取物;
(2)乙酸乙酯萃取物经正相硅胶柱色谱梯度洗脱,得到6个组分 A1~A6,A4组分经硅胶柱色谱分离洗脱,得到A41~A45五个组分,A43经Sephadex LH-20柱色谱分离,继续得到3个组分A431~A433,A432组分经半制备高效液相色谱分离得到二萜类化合物1和二萜类化合物2;A5组分经Sephadex LH-20柱色谱分离,继续得到5个组分A51~A55,A53组分经半制备高效液相色谱分离得到化合物二萜类化合物3;
所述正相硅胶柱色谱梯度洗脱采用石油醚和乙酸乙酯进行梯度洗脱,石油醚和乙酸乙酯的体积比为1:1~2;
所述A4组分经硅胶柱色谱分离洗脱采用二氯甲烷和甲醇洗脱,二氯甲烷和甲醇的体积比为8:1~1:3;
所述A432组分经半制备高效液相色谱分离采用55%甲醇-45%水;
所述A53组分经半制备高效液相色谱分离采用55%甲醇-45%水。
3.一种权利要求1所述三个具有激活潜伏HIV作用的二萜类成分的应用,其特征在于,在制备清除潜伏HIV病毒药物中的应用。
4.根据权利要求3所述的三个具有激活潜伏HIV作用的二萜类成分的应用,其特征在于,所述药物以二萜类化合物直接使用,或以药学上可接受的盐以及药物组合物的形式使用。
5.根据权利要求4所述的三个具有激活潜伏HIV作用的二萜类成分的应用,其特征在于,所述药物组合物含有质量分数为0.1~99%的二萜类化物,其余为药物学上可接受的、对人和动物无毒和惰性的可药用载体和/或赋形剂。
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN202210239249.2A CN114573600B (zh) | 2022-03-11 | 2022-03-11 | 四个具有激活潜伏hiv作用的二萜类成分的制备方法和应用 |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN202210239249.2A CN114573600B (zh) | 2022-03-11 | 2022-03-11 | 四个具有激活潜伏hiv作用的二萜类成分的制备方法和应用 |
Publications (2)
Publication Number | Publication Date |
---|---|
CN114573600A CN114573600A (zh) | 2022-06-03 |
CN114573600B true CN114573600B (zh) | 2024-03-12 |
Family
ID=81780497
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN202210239249.2A Active CN114573600B (zh) | 2022-03-11 | 2022-03-11 | 四个具有激活潜伏hiv作用的二萜类成分的制备方法和应用 |
Country Status (1)
Country | Link |
---|---|
CN (1) | CN114573600B (zh) |
Families Citing this family (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN115232143B (zh) * | 2022-07-18 | 2023-10-13 | 沈阳药科大学 | 黄芫花中制备的瑞香烷型二萜类化合物及其制备方法和应用 |
CN115785043B (zh) * | 2022-12-12 | 2024-03-12 | 山西大学 | 愈创木烷型倍半萜激活潜伏hiv的应用及其制备方法 |
Citations (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN110317209A (zh) * | 2019-07-31 | 2019-10-11 | 山西大学 | 二萜类化合物wikstroelide E及在制备清除潜伏HIV病毒的药物中的应用 |
-
2022
- 2022-03-11 CN CN202210239249.2A patent/CN114573600B/zh active Active
Patent Citations (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN110317209A (zh) * | 2019-07-31 | 2019-10-11 | 山西大学 | 二萜类化合物wikstroelide E及在制备清除潜伏HIV病毒的药物中的应用 |
Non-Patent Citations (2)
Title |
---|
Anti-HIV-1 tigliane diterpenoids from Excoecaria acertiflia Didr;Sheng-Zhuo Huang,等;《Fitoterapia》;第95卷;第39页化合物1 * |
Daphnane- and Tigliane-Type Diterpenoid Esters and Orthoesters from Pimelea elongata;Patricia Y. Hayes;《J. Nat. Prod.》;20100411;第73卷;1908页化合物5 * |
Also Published As
Publication number | Publication date |
---|---|
CN114573600A (zh) | 2022-06-03 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
CN114573600B (zh) | 四个具有激活潜伏hiv作用的二萜类成分的制备方法和应用 | |
CN110317209B (zh) | 二萜类化合物wikstroelide E及在制备清除潜伏HIV病毒的药物中的应用 | |
CN102727563B (zh) | 大戟属植物具有抗hiv潜伏作用的有效部位及其应用 | |
CN101669979B (zh) | 滨蒿提取物及其生产方法和应用 | |
CA2006263A1 (en) | Sulfated tannins and their salts | |
CN100497279C (zh) | 一类对映桉烷醇类倍半萜抑制乙肝病毒的医药用途 | |
CN112168951A (zh) | 茜草环肽化合物在制药中的应用 | |
CN113304165B (zh) | 单体化合物Ciliatoside A在制备乙型肝炎治疗药物中的用途 | |
CN115894418A (zh) | 蒙古蒿内酯a-f及其药物组合物和其制备方法与应用 | |
CN115785043B (zh) | 愈创木烷型倍半萜激活潜伏hiv的应用及其制备方法 | |
JPH05331061A (ja) | アポトーシス誘起剤 | |
CN108888628B (zh) | 一种人参皂苷GRh2在制备抗弓形虫复方制剂中的应用及其药物 | |
EP3398596A1 (en) | Ingenol compounds and use thereof in anti-hiv latency treatment | |
CN102584846B (zh) | 一种抗甲型h1n1流感病毒的莪术醇衍生物 | |
CN102727551B (zh) | 一种中药有效部位在抗hiv潜伏治疗上的应用 | |
CN100482217C (zh) | 2β-羟基冬青酸在制备抑制乙肝表面抗原药物中的应用 | |
CN1105238A (zh) | 抗aids病毒的药物组合物 | |
CN1935131B (zh) | 1β-羟基冬青酸用于制备抑制乙肝病毒的药物用途 | |
CN1303673A (zh) | 槐果碱作为制备柯萨奇b病毒性心肌炎病因治疗药物的用途及其制法 | |
JPH03271226A (ja) | 腎炎治療剤 | |
CN1927197A (zh) | 1β-氧代-5,11(13)-二烯桉烷-12-酸抑制乙肝病毒的医药用途 | |
CN1035767C (zh) | 新的青蒿素衍生物及其用途 | |
JPH11193242A (ja) | 抗インフルエンザウイルス剤 | |
CN1200708C (zh) | 四环香豆素类化合物在制备抗艾滋病毒类药物中的应用 | |
CN112107571A (zh) | 巨大戟二萜醇类化合物及其衍生物在抗hiv治疗上的应用 |
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 |