WO2016184361A1 - 抗丙肝病毒的核苷类化合物及其应用 - Google Patents

抗丙肝病毒的核苷类化合物及其应用 Download PDF

Info

Publication number
WO2016184361A1
WO2016184361A1 PCT/CN2016/082001 CN2016082001W WO2016184361A1 WO 2016184361 A1 WO2016184361 A1 WO 2016184361A1 CN 2016082001 W CN2016082001 W CN 2016082001W WO 2016184361 A1 WO2016184361 A1 WO 2016184361A1
Authority
WO
WIPO (PCT)
Prior art keywords
alkyl
substituted
unsubstituted
halogens
independently
Prior art date
Application number
PCT/CN2016/082001
Other languages
English (en)
French (fr)
Inventor
张所明
Original Assignee
上海唐润医药科技有限公司
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
Application filed by 上海唐润医药科技有限公司 filed Critical 上海唐润医药科技有限公司
Publication of WO2016184361A1 publication Critical patent/WO2016184361A1/zh

Links

Classifications

    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
    • C07H19/00—Compounds containing a hetero ring sharing one ring hetero atom with a saccharide radical; Nucleosides; Mononucleotides; Anhydro-derivatives thereof
    • C07H19/02—Compounds containing a hetero ring sharing one ring hetero atom with a saccharide radical; Nucleosides; Mononucleotides; Anhydro-derivatives thereof sharing nitrogen
    • C07H19/04—Heterocyclic radicals containing only nitrogen atoms as ring hetero atom
    • C07H19/06—Pyrimidine radicals
    • C07H19/10—Pyrimidine radicals with the saccharide radical esterified by phosphoric or polyphosphoric acids
    • C07H19/11—Pyrimidine radicals with the saccharide radical esterified by phosphoric or polyphosphoric acids containing cyclic phosphate
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
    • A61P1/16—Drugs for disorders of the alimentary tract or the digestive system for liver or gallbladder disorders, e.g. hepatoprotective agents, cholagogues, litholytics
    • 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
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
    • C07H19/00—Compounds containing a hetero ring sharing one ring hetero atom with a saccharide radical; Nucleosides; Mononucleotides; Anhydro-derivatives thereof
    • C07H19/02—Compounds containing a hetero ring sharing one ring hetero atom with a saccharide radical; Nucleosides; Mononucleotides; Anhydro-derivatives thereof sharing nitrogen
    • C07H19/04—Heterocyclic radicals containing only nitrogen atoms as ring hetero atom
    • C07H19/16—Purine radicals
    • C07H19/20—Purine radicals with the saccharide radical esterified by phosphoric or polyphosphoric acids
    • C07H19/213—Purine radicals with the saccharide radical esterified by phosphoric or polyphosphoric acids containing cyclic phosphate
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
    • C07H19/00—Compounds containing a hetero ring sharing one ring hetero atom with a saccharide radical; Nucleosides; Mononucleotides; Anhydro-derivatives thereof
    • C07H19/02—Compounds containing a hetero ring sharing one ring hetero atom with a saccharide radical; Nucleosides; Mononucleotides; Anhydro-derivatives thereof sharing nitrogen
    • C07H19/04—Heterocyclic radicals containing only nitrogen atoms as ring hetero atom
    • C07H19/23—Heterocyclic radicals containing two or more heterocyclic rings condensed among themselves or condensed with a common carbocyclic ring system, not provided for in groups C07H19/14 - C07H19/22

Definitions

  • the invention belongs to the field of antiviral compounds, and particularly relates to a class of anti-hepatitis C virus nucleoside compounds and applications thereof.
  • a virus is a non-cellular form of parasitic living organism composed of a nucleic acid molecule (DNA or RNA) and a protein or only a protein.
  • Hepatitis C virus HCV
  • HCV Hepatitis C virus
  • IFN interferon
  • HCV-infected patients have different responses to interferon therapy, the average response rate is less than 50%, and the recurrence rate after drug withdrawal is higher.
  • Nucleoside compounds have effects on various genotypes and have high gene blocking effects on viral resistance. If it can screen out the safe and effective compounds that patients can afford, it is likely to become an important hepatitis C treatment.
  • the present invention is directed to the technical problems in the prior art, and an object thereof is to provide a compound having the formula (I), and a pharmaceutically acceptable salt thereof,
  • B is a nucleoside base
  • Z is a connection key or CHR 4 ;
  • R is CH(R')C(Y)OR 5 ; when W is CHC(Y)OR 5 , R is H; R' is H, D or not a C 1-6 alkyl group or a C 3-8 cycloalkyl group substituted or substituted with 1 to 5 halogens;
  • Y is O or S
  • R 1 is H, D or CN
  • R 2 is H, D, X, CN, CHCH 2 , CCH or N 3 ;
  • Each R 3 is independently H, D, X, OH, N 3 , NR" 2 or C 1-6 alkyl, C 2-6 alkenyl or C 2 which is unsubstituted or substituted with 1 to 5 halogens. -6 alkynyl;
  • R 5 is C 1-6 alkyl or C 3-8 cycloalkyl which is unsubstituted or substituted by 1 to 5 halogen or C 1-6 alkoxy; each R a is independently hydrogen, X, CN , R", NR" 2 , OR” or CO 2 R", or R a on two adjacent carbon atoms forms a 3-8 carbon ring with the attached carbon atom;
  • Each R" is independently D or a C 1-6 alkyl group, a C 3-8 cycloalkyl group, a C 1-6 acyl group or a C 1-6 sulfonamide group which is unsubstituted or substituted with 1 to 5 halogens; Or two R" may form a 4-8 membered ring with the attached nitrogen atom; X is a halogen.
  • B 1 is N or NH
  • B 2 is N
  • B 3 is C or N, such that B forms a 6-membered aza-aryl ring
  • R p and R q are independently H, D, X, C 1-6 alkyl, C 2-6 alkenyl or C 2-6 alkynyl;
  • the pentose sugar is C-1 is as shown by 1' in the structural formula of pentose.
  • the 9-membered heterocyclic nitrogen ring is B 6 is N or CR c and the ninth element is B 5 .
  • R c is H, D, C 1-6 alkyl, C 2-6 alkenyl or C 2-6 alkynyl.
  • B 4 is N or CR c
  • B 5 is C or N
  • at least one of B 3 , B 4 and B 5 is N.
  • B is Preferably selected from:
  • R p and R q are each H, and B 4 is N or CR c .
  • the representative can be either a single button or a double button, with B being
  • the ring structure needs to satisfy B as the aromatic ring.
  • B is preferred It is also necessary to satisfy the B of the double ring structure as the aromatic ring.
  • R' is H, D or C 1-4 alkyl or C 3-6 cycloalkyl which is unsubstituted or substituted with 1 to 3 halogens; preferably, R' is H, D or unsubstituted or a C 1-2 alkyl group or a C 5-6 cycloalkyl group substituted by 1 to 2 halogens;
  • Each R 3 is independently H, D, X, OH, N 3 , NR" 2 or C 1-4 alkyl, C 2-4 alkenyl or C 2 which is unsubstituted or substituted with 1 to 3 halogens. -4 alkynyl; preferably, each R 3 is independently H, D, X, OH, N 3 , NR" 2 or C 1-2 alkyl unsubstituted or substituted with 1 to 2 halogens, C 2-3 alkenyl or C 2-3 alkynyl;
  • R 5 is C 1-4 alkyl or C 3-6 cycloalkyl which is unsubstituted or substituted by 1 to 3 halogen or C 1-4 alkoxy; preferably, R 5 is unsubstituted or a C 1-2 alkyl group or a C 5-6 cycloalkyl group substituted by 1 to 2 halogens or a C 1-2 alkoxy group;
  • Each R a is independently hydrogen, X, CN, R", NR" 2 , OR” or CO 2 R", or R a on two adjacent carbon atoms forms a 3 to 6 carbon ring with the attached carbon atom. , preferably a 5- to 6-membered carbocyclic ring, more preferably a 6-membered aromatic ring;
  • Each R" is independently D or a C 1-4 alkyl group which is unsubstituted or substituted with 1 to 3 halogens, a C 3-6 cycloalkyl group, a C 1-4 acyl group or a C 1-4 sulfonamide group; Or two R" may form a 4- to 6-membered ring with the attached nitrogen atom; preferably, each R" is independently D or a C 1-2 alkyl group which is unsubstituted or substituted with 1 to 2 halogens, C a 5-6 cycloalkyl group, a C 1-2 acyl group or a C 1-2 sulfonamide group; or two R" may form a 5- to 6-membered ring with a linked nitrogen atom;
  • R p and R q are independently H, D, X, C 1-4 alkyl, C 2-4 alkenyl or C 2-4 alkynyl, preferably R p and R q are independently H, D, X, C 1-2 alkyl, C 2-3 alkenyl or C 2-3 alkynyl;
  • R c is H, D, C 1-4 alkyl, C 2-4 alkenyl or C 2-4 alkynyl; preferably H, D, C 1-2 alkyl, C 2-3 alkenyl or C 2-3 alkynyl;
  • X is F or Cl.
  • Another object of the present invention is to provide a compound represented by the formula (I) for the preparation of an anti-HCV drug Application.
  • a further object of the invention is a method of treating hepatitis C disease comprising the step of administering to a hepatitis C patient an effective amount of a compound of formula (I).
  • the 2-amino-3-(2-hydroxyphenyl)propionic acid was used in place of 2-hydroxyphenylglycine, and the same as in Example 1, the compound 3 was obtained (yield: 70%).
  • Salicylaldehyde (1.22 g) was dissolved in 1,2-dichloroethane (20 mL), and L-alanine isopropyl ester hydrochloride (1.68 g), triethylamine (1.01 g), triacetyl was added.
  • Sodium oxyborohydride (4.2 g) and a few drops of acetic acid. Stir at room temperature until the reaction is complete. The reaction mixture was diluted with water, extracted with methylene chloride, dried over anhydrous magnesium sulfate.
  • L-Alanine isopropyl ester hydrochloride (1.68 g) and triethylamine (1.01 g) were dissolved in 20 mL of dichloromethane, cooled to 0 ° C, and then the above-prepared salicyl chloride dichloromethane was added dropwise. Solution. The reaction mixture was stirred overnight. Then, it was washed with 40 mL of water and 1N of hydrochloric acid, washed with water, dried over anhydrous sodium sulfate, filtered and evaporated to dryness.
  • the following target compounds can be obtained by the following synthesis methods using the following intermediate compounds and corresponding nucleosides;
  • HCV replicon transfected cells Huh7.5.1 cells transfected with HCV replicon (wild type 1b). The transfected cells were seeded in 96-well plates, 8000 cells per well, and cultured at 37 ° C, 5% CO 2 for 24 hours.
  • Sample treatment Different concentrations of Compound 1-4 samples were added to Huh7.5.1 cells transfected with HCV replicon, two replicate wells were set for each concentration, and no sample control wells were set. The test sample was started from the highest concentration tested, and the POD810 automatic microplate pretreatment system was added to the cells with different concentrations of the compound; the concentration was diluted 3 times; the culture was continued for 72 hours.
  • the fluorescence signal was measured by adding Cell Titer-fluor (Promega), and the obtained data (RFU) was calculated using the GraphPad Prism software to calculate the EC 50 of the compound.
  • Compounds 1 to 4 all showed activity for inhibiting the HCV Gt1b genotype, as shown in Table 1.
  • Table 1 EC 50 values of compounds 1 to 4 for HCV 1b genotype replicons
  • A indicates EC 50 ⁇ 1 ⁇ M
  • B indicates 1 ⁇ M ⁇ EC 50 ⁇ 10 ⁇ M.
  • the compounds of the invention having the general formula (I) have excellent anti-hepatitis virus activity, especially against hepatitis C virus.

Abstract

本发明公开了具有通式(I)所示的化合物，及其药物上可接受的盐，A为O或C＝CH2；B为核苷类碱基；Z为连接键或CHR4；当W为连接键或C(O)时，R为CH(R')C(Y)OR5；当W为CHC(Y)OR5时，R为H；Y为O或S；R1为H、D或CN；R2为H、D、X、CN、CH＝CH2、CCH或N3；各R3独立地为H、D、X、OH、N3、NR"2或未被取代的或被1～5个卤素取代的C1-6烷基、C2-6烯基或C2-6炔基；各Ra独立地为H、X、CN、R"、NR"2、OR"或CO2R"，或者相邻两个碳原子上的Ra与连接的碳原子形成3～8元碳环；各R"独立地为D、C1-6烷基、C3-8环烷基、C1-6酰基或C1-6磺酰胺基；或者两个R"可以与相连的氮原子形成4～8元环。本发明的通式(I)所示的化合物具有优异的抗丙肝病毒的效果。

Description

抗丙肝病毒的核苷类化合物及其应用 技术领域
本发明属于抗病毒化合物领域，具体涉及一类抗丙肝病毒的核苷类化合物及其应用。
背景技术
病毒(virus)是由一个核酸分子(DNA或RNA)与蛋白质构成或仅由蛋白质构成的非细胞形态的靠寄生生活的生命体。丙型肝炎病毒(HCV)，为单股正链RNA病毒，是输血后以及散发性肝炎的主要病原。目前公认对丙型肝炎抗病毒治疗有效药物是干扰素(IFN)，但HCV感染者对干扰素治疗的应答不一，平均应答率不足50％，且停药后复发率较高。FDA于2011年批准了两个NS3/4A丝氨酸蛋白酶抑制剂Telaprevir和Boceprevir上市，为丙型肝炎的治疗提供了新的有效方法。2013年FDA又批准了Sofosbuvir和Simeprevir上市。但是耐药性和毒副作用的出现，使得HCV病毒的治疗仍然需要新型药物。
核苷类化合物对各种基因型均有作用，而且对病毒的耐药性具有很高的基因屏蔽作用。若是能筛选出病人所能承受的安全有效的化合物，很有可能成为重要的丙肝治疗药物。
发明内容
本发明针对现有技术中的技术问题，目的在于提供一种具有通式(Ⅰ)所示的化合物，及其药物上可接受的盐，
Figure PCTCN2016082001-appb-000001
A为O或C＝CH2；
B为核苷类碱基；
Z为连接键或CHR4；
当W为连接键或C(O)时，R为CH(R')C(Y)OR5；当W为CHC(Y)OR5时，R 为H；R'为H、D或未被取代的或被1～5个卤素取代的C1-6烷基或C3-8环烷基；
Y为O或S；
R1为H、D或CN；
R2为H、D、X、CN、CHCH2、CCH或N3；
各R3独立地为H、D、X、OH、N3、NR"2或未被取代的或被1～5个卤素取代的C1-6烷基、C2-6烯基或C2-6炔基；
R4为氢、氧代(＝O)或未被取代的或被1～5个卤素取代的C1-6烷基、C3-8环烷基、C2-6烯基或C2-6炔基；
R5为未被取代的或被1～5个卤素或C1-6烷氧基取代的C1-6烷基或C3-8环烷基；各Ra独立地为氢、X、CN、R"、NR"2、OR"或CO2R"，或者相邻两个碳原子上的Ra与连接的碳原子形成3～8元碳环；
各R"独立地为D或未被取代的或被1～5个卤素取代的C1-6烷基、C3-8环烷基、C1-6酰基或C1-6磺酰胺基；或者两个R"可以与相连的氮原子形成4～8元环；X为卤素。
在本发明中，B为
Figure PCTCN2016082001-appb-000002
即通式(Ⅰ)变为通式(Ⅰa)，
Figure PCTCN2016082001-appb-000003
B1为N或NH，B2为N，B3为C或N，使得B形成一6元杂氮芳环；
当Rp与Rq不相连成环时，戊糖上的C-1与6元杂氮芳环上的B2连接形成核苷键，此时Rp和Rq独立地为H、D、X、C1-6烷基、C2-6烯基或C2-6炔基；
本发明中，所述的戊糖为
Figure PCTCN2016082001-appb-000004
C-1如戊糖结构式中的1'所示。
当Rp与Rq相连形成一未被取代或被1～2个Rc取代的5元杂氮芳环，且与6元杂氮芳环一起共同形成9元杂氮稠芳环时，戊糖上的C-1与9元杂氮稠芳环上的第9位元素连接形成核苷键；
在本发明中，9元杂氮稠芳环为
Figure PCTCN2016082001-appb-000005
B6为N或CRc，第9位元素为B5。
Rb独立地为H、D、氧代(＝O)、硫代(＝S)、NR"2或OR"；
Rc为H、D、C1-6烷基、C2-6烯基或C2-6炔基。
式(Ⅰa)中的B为
Figure PCTCN2016082001-appb-000006
优选为
Figure PCTCN2016082001-appb-000007
B4为N或CRc，B5为C或N，且B3、B4和B5至少有1个为N。
式(Ⅰa)中，B为
Figure PCTCN2016082001-appb-000008
优选地选自：
Figure PCTCN2016082001-appb-000009
Figure PCTCN2016082001-appb-000010
其中，Rp与Rq分别为H，B4为N或CRc。
在本发明中，
Figure PCTCN2016082001-appb-000011
代表既可以为单键，也可以为双键，以B为
Figure PCTCN2016082001-appb-000012
为例，环状结构需要满足B为芳香环为准。当B优选为
Figure PCTCN2016082001-appb-000013
亦需要满足双环结构的B为芳香环为准。
具体地，可优选地选自
Figure PCTCN2016082001-appb-000014
式(Ⅰa)中，
Figure PCTCN2016082001-appb-000015
可选自：
Figure PCTCN2016082001-appb-000016
式(Ⅰ)中
Figure PCTCN2016082001-appb-000017
选自：
Figure PCTCN2016082001-appb-000018
在本发明一较佳实施例中，
R'为H、D或未被取代的或被1～3个卤素取代的C1-4烷基或C3-6环烷基；优选地，R'为H、D或未被取代的或被1～2个卤素取代的C1-2烷基或C5-6环烷基；
各R3独立地为H、D、X、OH、N3、NR"2或未被取代的或被1～3个卤素取代的C1-4烷基、C2-4烯基或C2-4炔基；优选地，各R3独立地为H、D、X、OH、N3、NR"2或未被取代的或被1～2个卤素取代的C1-2烷基、C2-3烯基或C2-3炔基；
R4为氢、氧代(＝O)或未被取代的或被1～3个卤素取代的C1-4烷基、C3-6环烷基、C2-4烯基或C2-4炔基；优选地，R4为氢、氧代(＝O)或未被取代的或被1～2个卤素取代的C1-2烷基、C5-6环烷基、C2-3烯基或C2-3炔基；
R5为未被取代的或被1～3个卤素或C1-4烷氧基取代的C1-4烷基或C3-6环烷基；优选地，R5为未被取代的或被1～2个卤素或C1-2烷氧基取代的C1-2烷基或C5-6环烷基；
各Ra独立地为氢、X、CN、R"、NR"2、OR"或CO2R"，或者相邻两个碳 原子上的Ra与连接的碳原子形成3～6元碳环，优选地5～6元碳环，更优选地6元芳环；
各R"独立地为D或未被取代的或被1～3个卤素取代的C1-4烷基、C3-6环烷基、C1-4酰基或C1-4磺酰胺基；或者两个R"可以与相连的氮原子形成4～6元环；优选地，各R"独立地为D或未被取代的或被1～2个卤素取代的C1-2烷基、C5-6环烷基、C1-2酰基或C1-2磺酰胺基；或者两个R"可以与相连的氮原子形成5～6元环；
当Rp与Rq不相连成环时，Rp和Rq独立地为H、D、X、C1-4烷基、C2-4烯基或C2-4炔基，优选地Rp和Rq独立地为H、D、X、C1-2烷基、C2-3烯基或C2-3炔基；
Rc为H、D、C1-4烷基、C2-4烯基或C2-4炔基；优选地为H、D、C1-2烷基、C2-3烯基或C2-3炔基；
X为F或Cl。
在本发明中，具体地，
式(Ⅰa)中
Figure PCTCN2016082001-appb-000019
可选自：
Figure PCTCN2016082001-appb-000020
本发明的另一目的在于提供式(Ⅰ)中所示的化合物在制备抗丙肝病毒药 物中的应用。
本发明的又一目的在于一种治疗丙肝疾病的方法，包括给予丙肝患者施加有效量的式(Ⅰ)中所示的化合物的步骤。
本发明的通式(Ⅰ)所示的化合物的合成流程如下所示：
Figure PCTCN2016082001-appb-000021
具体实施方式
实施例1
Figure PCTCN2016082001-appb-000022
步骤一：中间体1A(2-羟基苯甘氨酸异丙酯)的合成
2-羟基苯甘氨酸2g(0.01197mol,1eq)溶在40mL的异丙醇中，滴入2eq的SOCl2(2.8g,0.239mol)升温回流过夜。母液旋干，加入1mol/L稀盐酸和甲基叔丁基醚萃取2次，水相用饱和碳酸氢钠调节pH到碱性，用乙酸乙酯萃取3次，乙酸乙酯层用盐水洗涤，无水硫酸钠干燥，浓缩得到1.1g粗品，柱层析纯化，得到500mg中间体1A。
1H NMR(400MHz,CDCl3)
Figure PCTCN2016082001-appb-000023
7.211(td,J＝7.2Hz,J＝1.2Hz,1H),7.068(dd,J＝7.2Hz,J＝1.2Hz,1H),6.874(dd,J＝8.0Hz,J＝0.8Hz,1H)，6.827(td,J＝7.6Hz,J＝1.2Hz,1H),5.072～5.134(m,1H),4.802(s,1H),1.282(d,J＝6.4Hz,3H),1.191(d,J＝6.4Hz,3H)。
步骤二：中间体1B的合成
2-羟基苯甘氨酸异丙酯(500mg,2.4mmol)溶入三氯氧磷(10mL)中，在100℃加热搅拌过夜。将该混合物浓缩干，用甲苯溶解，浓缩至干得中间体1B。
步骤三：化合物1的合成
将中间体1B溶解于四氢呋喃(4mL)中，加入(2'R)-2'-脱氧-2'-氟-2'-甲基脲 苷(250mg,0.95mmol)和N-甲基咪唑(1.26g,15.4mmol)，混合物室温搅拌过夜。将反应液倒入水中，用1N盐酸调节pH至～3，用二氯甲烷萃取。二氯甲烷层用0.5N盐酸洗，饱和盐水洗，无水硫酸钠干燥，减压旋干得到粗产物。该粗产物用制备液相色谱(水-乙腈含0.01％甲酸)纯化得到化合物1(70mg,收率14％)。
1HNMR(400MHz,CDCl3)
Figure PCTCN2016082001-appb-000024
9.56-9.46(m,1H),7.67-7.62(m,1H),7.40-7.32(m,3H),7.21-7.14(m,1H),7.07(q,J＝7.2Hz,1H),6.22-6.08(m,1H),5.55-5.38(m,2H),5.15-4.95(m,2H),4.73-4.42(m,2H),4.19-3.89(m,2H),1.42-1.35(dd,3H),1.34-1.26(m,3H),1.17-1.15(dd,3H)；ES-LC/MS：m/z 514.1(M+H)。
实施例2
Figure PCTCN2016082001-appb-000025
用原料异丁醇代替异丙醇，其他同实施例1，得化合物2(收率50％)。
1HNMR(400MHz,CDCl3)
Figure PCTCN2016082001-appb-000026
9.1-9.2(m,1H),7.67-7.64(m,1H),7.40-7.32(m,3H),7.20-7.16(m,1H),7.11-7.07(m,1H),6.22-6.19(m,1H),5.60-5.47(m,1H),5.42-5.32(m,1H),5.19-5.10(m,1H),4.73-4.42(m,2H),4.19-4.05(m,2H),3.96(d,J＝8.0Hz,2H),1.89-1.95(m,1H),1.44-1.34(dd,3H),0.84-0.91(m,6H)；ES-LC/MS：m/z 528.2(M+H)。
实施例3
Figure PCTCN2016082001-appb-000027
用原料2-氨基-3-(2-羟基苯基)丙酸代替2-羟基苯甘氨酸，其他同实施例1，得化合物3(收率：70％)
ES-LC/MS：m/z 528.2(M+H)。
实施例4
Figure PCTCN2016082001-appb-000028
步骤一：中间体4A的合成
将水杨醛(1.22g)溶解在1,2-二氯乙烷(20mL)中，加入L-丙氨酸异丙酯盐酸盐(1.68g)、三乙胺(1.01g)、三乙酰氧基硼氢化钠(4.2g)和几滴醋酸。室温下搅拌直到反应完全。反应混合物加水稀释，用二氯甲烷萃取，无水硫酸镁干燥、过滤、滤液减压浓缩至干、柱层析纯化得中间体4A(收率：62％)。
ES-LC/MS:m/z 238.2(M+H)。
步骤二：中间体4B的合成
用中间体4A代替2-羟基苯甘氨酸异丙酯，其他同实施例1的步骤二，得中间体4B。
步骤三：化合物4的合成
用中间体4B代替中间体1B，其他同实施例1的步骤三，得化合物4(收率：48％)。
1HNMR(400MHz,CDCl3)
Figure PCTCN2016082001-appb-000029
8.25(brs,1H),7.49(d,J＝8.4Hz,1H),7.32-7.28(m,1H),7.16-7.06(m,4H),7.11-7.07(m,1H),6.19(d,J＝18.8Hz,1H),5.54(d,J＝8.0Hz,1H),4.94-4.89(m,1H),4.51-4.46(m,3H),4.39-4.25(m,1H),4.12(d,J＝8.8Hz,2H),3.96(brd,1H),3.26(brs,1H),1.58(d,J＝7.2Hz,1H),1.41(d,J＝22.4Hz,3H),1.18(d,J＝6.4Hz,3H),1.05(d,J＝6.4Hz,3H)；ES-LC/MS：m/z 542.2(M+H)。
实施例5
Figure PCTCN2016082001-appb-000030
中间体5A
向水杨酸(1.38g,10mmol)的二氯甲烷(20mL)中加一滴DMF(N,N-二甲基 甲酰胺)，然后在0℃滴加草酰氯(12mmol)，2小时后，减压浓缩至干，然后溶于20mL二氯甲烷中。
将L-丙氨酸异丙酯盐酸盐(1.68g)和三乙胺(1.01g)溶于20mL二氯甲烷中，冷却至0℃，然后滴加上面制备好的水杨酰氯二氯甲烷溶液。反应混合物搅拌过夜。然后依次用40mL水、1N的盐酸洗涤，水洗涤，无水硫酸钠干燥，过滤，减压浓缩至干，硅胶柱层析纯化得中间体5A(收率76％)。
1H-NMR(400MHz,CDCl3)
Figure PCTCN2016082001-appb-000031
12.2(s,1H),7.40-7.48(m,2H),7.03(brs,1H),7.00(dd,J＝0.8,8.4Hz,1H),6.89(dt,J＝0.8,8.0Hz,1H),5.13(m,1H),4.72(m,1H),1.54(d,J＝7.2Hz,3H),1.34(d,J＝6.4Hz,3H),1.31(d,J＝6.4Hz,3H)。
中间体5B
中间体5A(251mg,1mmol)和三氯化磷(5mmol)的甲苯(5mL)溶液回流5小时，减压浓缩至干。加入5mL无水甲苯，减压浓缩至干得中间体5B。
化合物5
将中间体5B(～1mmol)溶解于四氢呋喃(4mL)中，加入(2'R)-2'-脱氧-2'-氟-2'-甲基脲苷(250mg,0.95mmol)和N-甲基咪唑(1.26g,15.4mmol)，混合物室温搅拌过夜。将反应液倒入水中，用1N盐酸调节pH至～3，用二氯甲烷萃取。二氯甲烷层用0.5N盐酸洗，饱和盐水洗，无水硫酸钠干燥，减压旋干得到粗产物。该粗产物用硅胶柱色谱(二氯甲烷-甲醇100:2至100:10)纯化得到化合物5(收率34％)。ES-LC/MS：m/z 556(M+H)。
以下列原料为起始原料，再根据实施例1、4、5的方法，可得到相应的中间体化合物
表1 中间体化合物的合成
Figure PCTCN2016082001-appb-000032
Figure PCTCN2016082001-appb-000033
以下列中间体化合物和相应核苷为原料，按照化合物1的合成方法可得到下列目标化合物
表2 目标化合物
Figure PCTCN2016082001-appb-000034
Figure PCTCN2016082001-appb-000035
效果实施例HCV复制子细胞抑制活性
按照文献(Science.1999Jul 2；285(5424):110-3以及J.Virol.2003,Mar； 77(5):3007-19)中所述方法，运用HCV基因型GT1b复制子细胞系统(稳定转染HCV 1b复制子的Huh7细胞)对化合物1～4进行HCV 1b复制子的抑制活性检测。
HCV复制子转染细胞：HCV复制子(野生型1b)转染的Huh7.5.1细胞。将转染细胞接种于96孔板中，8000细胞每孔，在37℃、5％CO2培养24小时。
样品处理：在HCV复制子转染的Huh7.5.1细胞中加入不同浓度的化合物1～4样品，每个浓度设二复孔，并设无样品对照孔。受试样品从受试最高浓度开始，用POD810全自动微孔板预处理系统加不同浓度化合物至细胞中；3倍稀释10个浓度；继续培养72小时。
化合物的活性及细胞毒性测定：
加入Cell Titer-fluor(Promega)测定荧光信号，获得的数据(RFU)用GraphPad Prism软件计算化合物的EC50。
化合物1～4都表现出抑制HCV Gt1b基因型的活性，具体见表1。
表1 化合物1～4对HCV 1b基因型复制子的EC50值
Figure PCTCN2016082001-appb-000036
备注：A表示EC50≤1μM；B表示1μM＜EC50≤10μM。
工业实用性
本发明的具有通式(I)的化合物具有优异的抗肝炎病毒的活性，尤其是针对丙型肝炎病毒。
Figure PCTCN2016082001-appb-000037

Claims (10)

  1. 具有通式(Ⅰ)所示的化合物，及其药物上可接受的盐，
    Figure PCTCN2016082001-appb-100001
    A为O或C＝CH2；
    B为核苷类碱基；
    Z为连接键或CHR4；
    当W为连接键或C(O)时，R为CH(R')C(Y)OR5；当W为CHC(Y)OR5时，R为H；R'为H、D或未被取代的或被1～5个卤素取代的C1-6烷基或C3-8环烷基；
    Y为O或S；
    R1为H、D或CN；
    R2为H、D、X、CN、CHCH2、CCH或N3；
    各R3独立地为H、D、X、OH、N3、NR"2或未被取代的或被1～5个卤素取代的C1-6烷基、C2-6烯基或C2-6炔基；
    R4为氢、氧代(＝O)或未被取代的或被1～5个卤素取代的C1-6烷基、C3-8环烷基、C2-6烯基或C2-6炔基；
    R5为未被取代的或被1～5个卤素或C1-6烷氧基取代的C1-6烷基或C3-8环烷基；各Ra独立地为氢、X、CN、R"、NR"2、OR"或CO2R"，或者相邻两个碳原子上的Ra与连接的碳原子形成3～8元碳环；
    各R"独立地为D或未被取代的或被1～5个卤素取代的C1-6烷基、C3-8环烷基、C1-6酰基或C1-6磺酰胺基；或者两个R"可以与相连的氮原子形成4～8元环；X为卤素。
  2. 如权利要求1所述的化合物，其特征在于，B为
    Figure PCTCN2016082001-appb-100002
    即通式(Ⅰ)变为通式(Ⅰa)，
    Figure PCTCN2016082001-appb-100003
    B1为N或NH，B2为N，B3为C或N，使得B形成一6元杂氮芳环；
    当Rp与Rq不相连成环时，戊糖上的C-1与6元杂氮芳环上的B2连接形成核苷键，此时Rp和Rq独立地为H、D、X、C1-6烷基、C2-6烯基或C2-6炔基；
    当Rp与Rq相连形成一未被取代或被1～2个Rc取代的5元杂氮芳环，且与6元杂氮芳环一起共同形成9元杂氮稠芳环时，戊糖上的C-1与9元杂氮稠芳环上的第9位元素连接形成核苷键；
    Rb独立地为H、D、氧代(＝O)、硫代(＝S)、NR"2或OR"；
    Rc为H、D、C1-6烷基、C2-6烯基或C2-6炔基。
  3. 如权利要求2所述的化合物，其特征在于，
    式(Ⅰa)中
    Figure PCTCN2016082001-appb-100004
    B4为N或CRc，B5为C或N，且B3、B4和B5至少有1个为N。
  4. 如权利要求2所述的化合物，其特征在于，
    式(Ⅰa)中，
    Figure PCTCN2016082001-appb-100005
    选自：
    Figure PCTCN2016082001-appb-100006
    Figure PCTCN2016082001-appb-100007
    其中，Rp与Rq分别为H，B4为N或CRc。
  5. 如权利要求4所述的化合物，其特征在于，
    式(Ⅰa)中
    Figure PCTCN2016082001-appb-100008
    选自：
    Figure PCTCN2016082001-appb-100009
  6. 如权利要求1所述的化合物，其特征在于，
    式(Ⅰa)中，
    Figure PCTCN2016082001-appb-100010
    选自
    Figure PCTCN2016082001-appb-100011
  7. 如权利要求2所述的化合物，其特征在于，
    式(Ⅰ)中
    Figure PCTCN2016082001-appb-100012
    选自：
    Figure PCTCN2016082001-appb-100013
  8. 如权利要求2所述的化合物，其特征在于，
    R'为H、D或未被取代的或被1～3个卤素取代的C1-4烷基或C3-6环烷基；优选地，R'为H、D或未被取代的或被1～2个卤素取代的C1-2烷基或C5-6环烷基；
    各R3独立地为H、D、X、OH、N3、NR"2或未被取代的或被1～3个卤素取代的C1-4烷基、C2-4烯基或C2-4炔基；优选地，各R3独立地为H、D、X、OH、N3、NR"2或未被取代的或被1～2个卤素取代的C1-2烷基、C2-3烯基或 C2-3炔基；
    R4为氢、氧代(＝O)或未被取代的或被1～3个卤素取代的C1-4烷基、C3-6环烷基、C2-4烯基或C2-4炔基；优选地，R4为氢、氧代(＝O)或未被取代的或被1～2个卤素取代的C1-2烷基、C5-6环烷基、C2-3烯基或C2-3炔基；
    R5为未被取代的或被1～3个卤素或C1-4烷氧基取代的C1-4烷基或C3-6环烷基；优选地，R5为未被取代的或被1～2个卤素或C1-2烷氧基取代的C1-2烷基或C5-6环烷基；
    各Ra独立地为氢、X、CN、R"、NR"2、OR"或CO2R"，或者相邻两个碳原子上的Ra与连接的碳原子形成3～6元碳环，优选地5～6元碳环，更优选地6元芳环；
    各R"独立地为D或未被取代的或被1～3个卤素取代的C1-4烷基、C3-6环烷基、C1-4酰基或C1-4磺酰胺基；或者两个R"可以与相连的氮原子形成4～6元环；优选地，各R"独立地为D或未被取代的或被1～2个卤素取代的C1-2烷基、C5-6环烷基、C1-2酰基或C1-2磺酰胺基；或者两个R"可以与相连的氮原子形成5～6元环；
    当Rp与Rq不相连成环时，Rp和Rq独立地为H、D、X、C1-4烷基、C2-4烯基或C2-4炔基，优选地Rp和Rq独立地为H、D、X、C1-2烷基、C2-3烯基或C2-3炔基；
    Rc为H、D、C1-4烷基、C2-4烯基或C2-4炔基；优选地为H、D、C1-2烷基、C2-3烯基或C2-3炔基；
    X为F或Cl。
  9. 权利要求1所述的化合物在制备抗丙肝病毒药物中的应用。
  10. 一种治疗丙肝疾病的方法，包括给予丙肝患者施加有效量的权利要求1所述的化合物的步骤。
PCT/CN2016/082001 2015-05-15 2016-05-13 抗丙肝病毒的核苷类化合物及其应用 WO2016184361A1 (zh)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
CN2015102489962 2015-05-15
CN201510248996.2A CN106279322B (zh) 2015-05-15 2015-05-15 抗丙肝病毒的核苷类化合物及其应用

Publications (1)

Publication Number Publication Date
WO2016184361A1 true WO2016184361A1 (zh) 2016-11-24

Family

ID=57319427

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/CN2016/082001 WO2016184361A1 (zh) 2015-05-15 2016-05-13 抗丙肝病毒的核苷类化合物及其应用

Country Status (2)

Country Link
CN (1) CN106279322B (zh)
WO (1) WO2016184361A1 (zh)

Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN114262348A (zh) * 2020-09-16 2022-04-01 上海本仁科技有限公司 环状核苷磷酸酯类化合物及其应用
US11697666B2 (en) 2021-04-16 2023-07-11 Gilead Sciences, Inc. Methods of preparing carbanucleosides using amides
US11767337B2 (en) 2020-02-18 2023-09-26 Gilead Sciences, Inc. Antiviral compounds

Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN1290707A (zh) * 2000-10-09 2001-04-11 清华大学 核苷5’－硫代磷酰氨基酸酯化合物
CA2749394A1 (en) * 2009-01-09 2010-07-15 University College Of Cardiff Consultants Limited Phosphoramidate derivatives of guanosine nucleoside compounds for treatment of viral infections
CN103421068A (zh) * 2012-05-17 2013-12-04 世方药业（杭州）有限公司 具有抗病毒活性的杂芳基磷酰胺类化合物

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN1290707A (zh) * 2000-10-09 2001-04-11 清华大学 核苷5’－硫代磷酰氨基酸酯化合物
CA2749394A1 (en) * 2009-01-09 2010-07-15 University College Of Cardiff Consultants Limited Phosphoramidate derivatives of guanosine nucleoside compounds for treatment of viral infections
CN103421068A (zh) * 2012-05-17 2013-12-04 世方药业（杭州）有限公司 具有抗病毒活性的杂芳基磷酰胺类化合物

Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US11767337B2 (en) 2020-02-18 2023-09-26 Gilead Sciences, Inc. Antiviral compounds
CN114262348A (zh) * 2020-09-16 2022-04-01 上海本仁科技有限公司 环状核苷磷酸酯类化合物及其应用
US11697666B2 (en) 2021-04-16 2023-07-11 Gilead Sciences, Inc. Methods of preparing carbanucleosides using amides

Also Published As

Publication number Publication date
CN106279322B (zh) 2019-07-30
CN106279322A (zh) 2017-01-04

Similar Documents

Publication Publication Date Title
CN104395330B (zh) 尿嘧啶基螺氧杂环丁烷核苷
CA2899763C (en) Nucleoside phosphoramidate compound and use thereof
ES2470568T3 (es) Inhibidores macroc�clicos del virus de la hepatitis C
JP2008533057A (ja) 抗ウィルス剤の機能を有する両複素環連結化合物、その応用および当該化合物を含有する組成物
JP2009525287A (ja) ウイルスポリメラーゼインヒビター
CN104672288A (zh) 一种氘代索菲布韦及其用途
CN105732468B (zh) 一种n’-(2-(1h-吲哚-3-基)乙酰基)芳酰肼类化合物及其制备方法和用途
WO2016184361A1 (zh) 抗丙肝病毒的核苷类化合物及其应用
Shan et al. A novel class of geldanamycin derivatives as HCV replication inhibitors targeting on Hsp90: synthesis, structure–activity relationships and anti-HCV activity in GS4. 3 replicon cells
WO2017162204A1 (zh) 苯并呋喃类似物制备方法及其中间体和晶型
CA2871324A1 (en) Substituted acyloxyamidines as hcv ns3/4a inhibitors
Li et al. Synthesis and biological evaluation of sophocarpinic acid derivatives as anti-HCV agents
CN115504940A (zh) 一种酰胺类化合物、其制备方法和制药用途
CN105051036B (zh) 黄酮类或异黄酮类化合物及其用途
CN112079785A (zh) 一种新型抗流感病毒奥司他韦衍生物、其制备方法及用途
CN106967141B (zh) 核苷氨基磷酸酯化合物及其医药组合物和用途
CN107011206B (zh) 一种丙烯酰胺类化合物、其制备方法及医药用途
WO2020151296A1 (zh) 双核苷酸前体药物及其制备方法
Girijavallabhan et al. 2′-Modified Guanosine Analogs for the Treatment of HCV
CN105461774B (zh) 索非布韦的制备方法
CN112679489B (zh) N-磺酰基杂环衍生物及其制药用途
Zych et al. The effect of substitution patterns on the release rates of opioid peptides DADLE and [Leu5]-enkephalin from coumarin prodrug moieties
CN112574127B (zh) 喹唑啉类化合物及其制备方法、药物组合物及其应用
CN110092791B (zh) 5,6-吲哚并二恶烷类衍生物及其制备方法和应用
CN105985356B (zh) 咪唑[2,1-b]噻唑衍生物及其制备方法和用途

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: 16795848

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: 16795848

Country of ref document: EP

Kind code of ref document: A1