CN101302182A - Virus polymerase inhibitors - Google Patents
Virus polymerase inhibitors Download PDFInfo
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- CN101302182A CN101302182A CNA2008101085783A CN200810108578A CN101302182A CN 101302182 A CN101302182 A CN 101302182A CN A2008101085783 A CNA2008101085783 A CN A2008101085783A CN 200810108578 A CN200810108578 A CN 200810108578A CN 101302182 A CN101302182 A CN 101302182A
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- C07D333/00—Heterocyclic compounds containing five-membered rings having one sulfur atom as the only ring hetero atom
- C07D333/50—Heterocyclic compounds containing five-membered rings having one sulfur atom as the only ring hetero atom condensed with carbocyclic rings or ring systems
- C07D333/52—Benzo[b]thiophenes; Hydrogenated benzo[b]thiophenes
- C07D333/62—Benzo[b]thiophenes; Hydrogenated benzo[b]thiophenes with hetero atoms or with carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, e.g. ester or nitrile radicals, directly attached to carbon atoms of the hetero ring
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- C07D209/00—Heterocyclic compounds containing five-membered rings, condensed with other rings, with one nitrogen atom as the only ring hetero atom
- C07D209/02—Heterocyclic compounds containing five-membered rings, condensed with other rings, with one nitrogen atom as the only ring hetero atom condensed with one carbocyclic ring
- C07D209/04—Indoles; Hydrogenated indoles
- C07D209/08—Indoles; Hydrogenated indoles with only hydrogen atoms or radicals containing only hydrogen and carbon atoms, directly attached to carbon atoms of the hetero ring
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- C07D—HETEROCYCLIC COMPOUNDS
- C07D209/00—Heterocyclic compounds containing five-membered rings, condensed with other rings, with one nitrogen atom as the only ring hetero atom
- C07D209/02—Heterocyclic compounds containing five-membered rings, condensed with other rings, with one nitrogen atom as the only ring hetero atom condensed with one carbocyclic ring
- C07D209/04—Indoles; Hydrogenated indoles
- C07D209/30—Indoles; Hydrogenated indoles with hetero atoms or with carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, directly attached to carbon atoms of the hetero ring
-
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- C07D—HETEROCYCLIC COMPOUNDS
- C07D209/00—Heterocyclic compounds containing five-membered rings, condensed with other rings, with one nitrogen atom as the only ring hetero atom
- C07D209/02—Heterocyclic compounds containing five-membered rings, condensed with other rings, with one nitrogen atom as the only ring hetero atom condensed with one carbocyclic ring
- C07D209/04—Indoles; Hydrogenated indoles
- C07D209/30—Indoles; Hydrogenated indoles with hetero atoms or with carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, directly attached to carbon atoms of the hetero ring
- C07D209/42—Carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, e.g. ester or nitrile radicals
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- C07D—HETEROCYCLIC COMPOUNDS
- C07D401/00—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom
- C07D401/02—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom containing two hetero rings
- C07D401/04—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom containing two hetero rings directly linked by a ring-member-to-ring-member bond
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- C07D403/00—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00
- C07D403/02—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00 containing two hetero rings
- C07D403/04—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00 containing two hetero rings directly linked by a ring-member-to-ring-member bond
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- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D405/00—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom
- C07D405/02—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom containing two hetero rings
- C07D405/04—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom containing two hetero rings directly linked by a ring-member-to-ring-member bond
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- C07—ORGANIC CHEMISTRY
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- C07D409/00—Heterocyclic compounds containing two or more hetero rings, at least one ring having sulfur atoms as the only ring hetero atoms
- C07D409/02—Heterocyclic compounds containing two or more hetero rings, at least one ring having sulfur atoms as the only ring hetero atoms containing two hetero rings
- C07D409/04—Heterocyclic compounds containing two or more hetero rings, at least one ring having sulfur atoms as the only ring hetero atoms containing two hetero rings directly linked by a ring-member-to-ring-member bond
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- C07D—HETEROCYCLIC COMPOUNDS
- C07D413/00—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and oxygen atoms as the only ring hetero atoms
- C07D413/02—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and oxygen atoms as the only ring hetero atoms containing two hetero rings
- C07D413/04—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and oxygen atoms as the only ring hetero atoms containing two hetero rings directly linked by a ring-member-to-ring-member bond
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- C—CHEMISTRY; METALLURGY
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- C07D—HETEROCYCLIC COMPOUNDS
- C07D417/00—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for by group C07D415/00
- C07D417/02—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for by group C07D415/00 containing two hetero rings
- C07D417/04—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for by group C07D415/00 containing two hetero rings directly linked by a ring-member-to-ring-member bond
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- C—CHEMISTRY; METALLURGY
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- C07D—HETEROCYCLIC COMPOUNDS
- C07D471/00—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, at least one ring being a six-membered ring with one nitrogen atom, not provided for by groups C07D451/00 - C07D463/00
- C07D471/02—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, at least one ring being a six-membered ring with one nitrogen atom, not provided for by groups C07D451/00 - C07D463/00 in which the condensed system contains two hetero rings
- C07D471/04—Ortho-condensed systems
Abstract
An isomer, enantiomer, diastereoisomer, or tautomer of a compound, represented by formula (I):wherein:A is O, S, NR<1>, or CR1, wherein R1 is defined herein; represents either a single or a double bond;R2 is selected from: H, halogen, R21, OR21, SR21, COOR21, SO2N(R<22>)2, N(R<22>)2, , CON(R<22>)2, NR<22>C(O)R<22> or NR<22>C(O)NR<22> wherein R<21> and each R<22> is defined herein;B is NR<3> or CR<3>, with the proviso that one of A or B is either CR<1> or CR<3>, wherein R<3> is defined herein;K is N or CR<4>, wherein R<4> is defined herein;L is N or CR<5>, wherein R<5> has the same definition as R<4> defined above;M is N or CR<7>, wherein R<7> has the same definition as R<4> defined above;Y<1> is O or S;Z is N(R<6a>)R<6> or OR<6>, wherein R<6a> is H or alkyl or NR<61>R<62> wherein R<61> and R<62> are defined herein;a salt or a derivative thereof, as an inhibitor of HCV NS<5B> polymerase.
Description
The application is a Chinese patent application (denomination of invention: viral polymerase inhibitors; The applying date: on July 18th, 2002; Application number: dividing an application 02818797.0).
Technical field
The present invention relates to those varial polymerases of RNA RNA-dependent polysaccharase, particularly flaviviridae, especially the inhibitor of HCV polysaccharase.
Background technology
It is estimated that the hepatitis C virus (HCV) of about 30000 examples of the annual kainogenesis of the U.S. infects (Kolykhalov, A.A.; Mihalik, K.; Feinstone, S.M.; Rice, C.M.; 2000; J.Virol.74:2046-2051).HCV is difficult for being removed by host's immune defense; Nearly HCV the infected of 85% transfers chronic infection to.A lot of this persistent infections cause chronic hepatopathy, comprise liver cirrhosis and hepatocellular carcinoma (Hoofnagle, J.H.; 1997; Hepatology 26:15S-20S).According to estimates, 1.7 hundred million HCV carrier are arranged worldwide, and the late period hepatopathy relevant with HCV is the major cause that causes liver transplantation now.Only in the U.S., hepatitis C just causes 8000～10000 people's death every year.If effectively do not intervene, expection this number in 10～20 years futures will reach now three times.There is not to prevent the vaccine of HCV infection at present.Adopting Interferon, rabbit or Interferon, rabbit and ribavirin is the therapy of present unique approval to chronic infection person's continued treatment, but this also only obtains lasting reaction (Lindsay, K.L. in less than 50% case; 1997; Hepatology 26:71S-77S, and Reichard, O.; Schvarcz, R.; Weiland, O.; 1997 Hepatology 26:108S-111S).
HCV belongs to flaviviridae, Hepacivirus, and it comprises that three belong to (Rice, C.M. for parcel film positive chain RNA virus; 1996; " Flaviviridae:the viruses and their replication "; Pp.931-960 in Fields Virology; Fields, B.N.; Knipe, D.M.; Howley, P.M. (eds.); Lippincott-Raven Publishers, Philadelphia Pa.).The 9.6 kb genomes of HCV are made up of the non-translational region (NTR) of lateral long open reading frame (ORF) and 5 ' and 3 ' flank thereof.The length of HCV 5 ' NTR is 341 Nucleotide, and serves as internal ribosome entry site (Lemon, the S.H. of cap-dependent/non-dependent translation initiation; Honda, M.; 1997; Semin.Virol.8:274-288).The HCV polyprotein is in translation or be cracked at least 10 peptide species (Reed, K.E. afterwards; Rice, C.M.; 1999; Curr.Top.Microbiol.Immunol.242:55-84).Structural protein are produced by the signal peptidase of polyprotein N-end.Two kinds of virus protease mediation downstream cracking produce unstructuredness (NS) albumen, and these albumen can be used as the integral part of HCV rna replicon enzyme.The C-that NS2-3 proteolytic enzyme is crossed over NS2 holds the N-of a half-sum NS3 to hold 1/3rd, and the cis cracking in catalyzing N S2/3 site.The same section of NS3 is also encoded and can be carried out cracked NS3-4A serine protease catalytic domain in site, four downstreams.Between various HCV isolates, the C-of NS3 end 2nd/3rd, high conservative, have RNA-combination, RNA-pungency NTPase and rna helicase activity.Although NS4B and NS5A phosphorprotein also may be the integral parts of replicative enzyme, its concrete effect is not clear.C-end polyprotein split product, NS5B is the extension subunit of HCV replicative enzyme, has RNA-RNA-dependent polysaccharase (RdRp) active (Behrens, S.E.; Tomei, L.; DeFrancesco, R.; 1996; EMBO J.15:12-22; And Lohmann, V.;
F.; Herian, U.; Bartenschlager, R.; 1997; J.Virol.71:8416-8428).Verified recently, the active sudden change of destruction NS5B can be eliminated RNA infectivity (Kolykhalov, the A.A. in chimpanzee (chimp) model; Mihalik, K.; Feinstone, S.M.; Rice, C.M.; 2000; J.Virol.74:2046-2051).
Be badly in need of new the treating of exploitation at present, and the necessary specific function of virus replication is a target the most attractive in the drug development with specific anti-HCV.Do not have RNA RNA-dependent polysaccharase in the Mammals, and this enzyme is that the necessary fact of virus replication hints that the NS5B polysaccharase is the dreamboat of anti-HCV therapy.
WO 00/06529 has reported the NS5B inhibitor, and it is alpha, gamma-two ketone acid.WO 00/13708, and WO 00/10573, and WO 00/18231, reaches WO 01/47883 and also reported the NS5B inhibitor, and recommend to be used for the treatment of HCV.
Summary of the invention
Therefore, the purpose of this invention is to provide and a series ofly have improved anti-HCV polysaccharase and suppress active new compound.
First aspect the invention provides compound and isomer thereof shown in the following formula I, enantiomer, and diastereomer, or tautomer:
In the formula:
A is O, S, NR
1, or CR
1, R wherein
1Be selected from: H, (C
1-6) alkyl, it is chosen wantonly and is replaced by following groups:
-halogen, OR
11, SR
11Or N (R
12)
2, R wherein
11And each R
12Be H independently, (C
1- 6) alkyl, (C
3-7) cycloalkyl, (C
1-6) alkyl-(C
3-7) cycloalkyl, (C
1-6) alkyl-aryl or (C
1-6) alkyl-Het, described aryl or Het are optional by R
10Replace; Perhaps
Two R
12, and be attached on the nitrogen that they connect by covalent bonds together, form 5,6 or the saturated heterocycle of 7-person;
-----represent singly-bound or two key;
R
2Be selected from: H, halogen, R
21, OR
21, SR
21, COOR
21, SO
2N (R
22)
2, N (R
22)
2, CON (R
22)
2, NR
22C (O) R
22Or NR
22C (O) NR
22, R wherein
21And each R
22Be H independently, (C
1-6) alkyl, haloalkyl, (C
2-6) alkenyl, (C
3-7) cycloalkyl, (C
2-6) alkynyl, (C
5-7) cycloalkenyl group, 6 or 10-person's aryl or Het, described R
21And R
22Optional by R
20Replace perhaps two R
22Be combined together to form 5,6 or the saturated heterocycle of 7-person with its nitrogen that is connected;
R wherein
10And R
20Respectively do for oneself:
-1～4 substituting groups are selected from: halogen, OPO
3H, NO
2, cyano group, azido-,
C (=NH) NH
2, C (=NH) NH (C
1-6) alkyl or C (=NH) NHCO (C
1-6) alkyl; Perhaps-1～4 substituting group is selected from:
A) (C
1-6) alkyl or haloalkyl, (C
3-7) cycloalkyl, optional comprise 1 or 2 heteroatomic C
3-7Spiro cycloalkyl group, (C
2-6) alkenyl, (C
3-6) cycloalkenyl group, (C
2-8) alkynyl, (C
1-6) alkyl-(C
3-7) cycloalkyl, all these groups are all randomly by R
150Replace;
B) OR
104, R wherein
104Be H, (C
1-6Alkyl), (C
3-7) cycloalkyl, or (C
1-6) alkyl-(C
3-7) cycloalkyl, aryl, Het, (C
1-6Alkyl) aryl or (C
1-6Alkyl) Het, described alkyl, cycloalkyl, aryl, Het, (C
1-6Alkyl) aryl or (C
1-6Alkyl) Het is all randomly by R
150Replace;
C) OCOR
105, R wherein
105Be (C
1-6) alkyl, (C
3-7) cycloalkyl, (C
1-6) alkyl-(C
3-7) cycloalkyl, Het, (C
1-6Alkyl) aryl or (C
1-6Alkyl) Het, described alkyl, cycloalkyl, aryl, Het, (C
1-6Alkyl) aryl or (C
1-6Alkyl) Het is all randomly by R
150Replace;
D) SR
108, SO
2N (R
108)
2Or SO
2N (R
108) C (O) R
108, each R wherein
108Be H independently, (C
1-6) alkyl, (C
3-7) cycloalkyl or (C
1-6) alkyl-(C
3-7) cycloalkyl, aryl, Het, (C
1-6Alkyl) aryl or (C
1-6Alkyl) Het, perhaps two R
108Combine formation 5,6 or the saturated heterocycle of 7-person, described alkyl, cycloalkyl, aryl, Het, (C by the connected nitrogen of covalent linkage
1-6Alkyl) aryl or (C
1-6Alkyl) Het or heterocycle are all optional by R
150Replace;
E) NR
111R
112, R wherein
111Be H, (C
1-6) alkyl, (C
3-7) cycloalkyl or (C
1-6) alkyl-(C
3-7) cycloalkyl, aryl, Het, (C
1-6Alkyl) aryl or (C
1-6Alkyl) Het, and R
112Be H, CN, (C
1-6) alkyl, (C
3-7) cycloalkyl or (C
1-6) alkyl-(C
3-7) cycloalkyl, aryl, Het, (C
1-6Alkyl) aryl, (C
1-6Alkyl) Het, COOR
115Or SO
2R
115' R wherein
115Be (C
1-6) alkyl, (C
3-7) cycloalkyl, or (C
1-6) alkyl-(C
3-7) cycloalkyl, aryl, Het, (C
1-6Alkyl) aryl or (C
1-6Alkyl) Het, perhaps two R
111And R
112Be combined together to form 5,6 or the saturated heterocycle of 7-person by the connected nitrogen of covalent linkage, described alkyl, cycloalkyl, aryl, Het, (C
1-6Alkyl) aryl or (C
1-6Alkyl) Het, or heterocycle is all randomly by R
150Replace;
F) NR
116COR
117, R wherein
116And R
117The H that respectively does for oneself, (C
1-6) alkyl, (C
3-7) cycloalkyl, (C
1-6) alkyl-(C
3-7) cycloalkyl, aryl, Het, (C
1-6Alkyl) aryl or (C
1-6Alkyl) Het, described (C
1-6) alkyl, (C
3-7) cycloalkyl, (C
1-6) alkyl-(C
3-7) cycloalkyl, aryl, Het, (C
1-6Alkyl) aryl or (C
1-6Alkyl) Het is all randomly by R
150Replace;
G) NR
118CONR
119R
120, R wherein
118, R
119And R
120The H that respectively does for oneself, (C
1-6) alkyl, (C
3-7) cycloalkyl, (C
1-6) alkyl-(C
3-7) cycloalkyl, aryl, Het, (C
1-6Alkyl) aryl or (C
1-6Alkyl) Het, perhaps R
118By covalent linkage and R
119In conjunction with and the nitrogen that is connected with them combine, form 5,6 or the saturated heterocycle of 7-person;
Perhaps R
119And R
120The nitrogen coupled by covalent linkage combines, formation 5,6 or the saturated heterocycle of 7-person, described alkyl, cycloalkyl, (C
1-6) alkyl-(C
3-7) cycloalkyl, aryl, Het, (C
1-6Alkyl) aryl or (C
1-6Alkyl) Het or heterocycle are all randomly by R
150Replace;
H) NR
121COCOR
122, R wherein
121And R
122The H that respectively does for oneself, (C
1-6) alkyl, (C
3-7) cycloalkyl, (C
1-6) alkyl-(C
3-7) cycloalkyl, 6-or 10-person's aryl, Het, (C
1-6Alkyl) aryl or (C
1-6Alkyl) Het, described alkyl, cycloalkyl, alkyl-cycloalkyl, aryl, Het, (C
1-6Alkyl) aryl or (C
1-6Alkyl) Het is all randomly by R
150Replace;
Perhaps R
122Be OR
123Or N (R
124)
2, R wherein
123And each R
124Be H independently, (C
1-6Alkyl), (C
3-7) cycloalkyl, (C
1-6) alkyl-(C
3-7) cycloalkyl, aryl, Het, (C
1-6Alkyl) aryl or (C
1-6Alkyl) Het, perhaps R
124Be OH or O (C
1-6Alkyl), perhaps two R
124By covalent bonds together, formation 5,6 or the saturated heterocycle of 7-person, described alkyl, cycloalkyl, alkyl-cycloalkyl, aryl, Het, (C
1-6Alkyl) aryl or (C
1-6Alkyl) Het and heterocycle are all randomly by R
150Replace;
I) COR
127, R wherein
127Be H, (C
1-6) alkyl, (C
3-7) cycloalkyl or (C
1-6) alkyl-(C
3-7) cycloalkyl, aryl, Het, (C
1-6Alkyl) aryl or (C
1-6Alkyl) Het, described alkyl, cycloalkyl, aryl, Het, (C
1-6Alkyl) aryl or (C
1-6Alkyl) Het is all randomly by R
150Replace;
J) COOR
128, R wherein
128Be H, (C
1-6) alkyl, (C
3-7) cycloalkyl, (C
1-6) alkyl-(C
3-7) cycloalkyl, aryl, Het, (C
1-6Alkyl) aryl or (C
1-6Alkyl) Het, described (C
1-6) alkyl, (C
3-7) cycloalkyl, or (C
1-6) alkyl-(C
3-7) cycloalkyl, aryl, Het, (C
1-6Alkyl) aryl and (C
1-6Alkyl) Het is all randomly by R
150Replace;
K) CONR
129R
130, R wherein
129And R
130Be H independently, (C
1-6) alkyl, (C
3-7) cycloalkyl, (C
1-6) alkyl-(C
3-7) cycloalkyl, aryl, Het, (C
1-6Alkyl) aryl or (C
1-6Alkyl) Het, perhaps two R
129And R
130The nitrogen coupled by covalent linkage combines, formation 5,6 or the saturated heterocycle of 7-person, described alkyl, cycloalkyl, alkyl-cycloalkyl, aryl, Het, (C
1-6Alkyl) aryl, (C
1-6Alkyl) Het and heterocycle are all randomly by R
150Replace;
L) aryl, Het, (C
1-6Alkyl) aryl or (C
1-6Alkyl) Het, all these groups are all randomly by R
150Replace; R wherein
150Be defined as follows:
-1～3 substituting groups are selected from: halogen, OPO
3H, NO
2, cyano group, azido-, C (=NH) NH
2, C (=NH) NH (C
1-6) alkyl or C (=NH) NHCO (C
1-6) alkyl; Perhaps
-1～3 substituting groups are selected from:
A) (C
1-6) alkyl or haloalkyl, (C
3-7) cycloalkyl, optional comprise 1 or 2 heteroatomic C
3-7Spiro cycloalkyl group, (C
2-6) alkenyl, (C
2-8) alkynyl, (C
1-6) alkyl-(C
3-7) cycloalkyl, all these groups are all optional by R
160Replace;
B) OR
104, R wherein
104Be H, (C
1-6Alkyl), (C
3-7) cycloalkyl, (C
1-6) alkyl-(C
3-7) cycloalkyl, aryl, Het, (C
1-6Alkyl) aryl or (C
1-6Alkyl) Het, described alkyl, cycloalkyl, aryl, Het, (C
1-6Alkyl) aryl or (C
1-6Alkyl) Het is all optional by R
160Replace;
C) OCOR
105, R wherein
105Be (C
1-6) alkyl, (C
3-7) cycloalkyl, (C
1-6) alkyl-(C
3-7) cycloalkyl, Het, (C
1-6Alkyl) aryl or (C
1-6Alkyl) Het, described alkyl, cycloalkyl, aryl, Het, (C
1-6Alkyl) aryl or (C
1-6Alkyl) Het is all optional by R
160Replace;
D) SR
108, SO
2N (R
108)
2Or SO
2N (R
108) C (O) R
108, each R wherein
108Be H independently, (C
1-6) alkyl, (C
3-7) cycloalkyl or (C
1-6) alkyl-(C
3-7) cycloalkyl, aryl, Het, (C
1-6Alkyl) aryl or (C
1-6Alkyl) Het, perhaps two R
108The nitrogen coupled by covalent linkage combines, formation 5,6 or the saturated heterocycle of 7-person, described alkyl, cycloalkyl, aryl, Het, (C
1-6Alkyl) aryl or (C
1-6Alkyl) Het or heterocycle are all optional by R
160Replace;
E) NR
111R
112, R wherein
111Be H, (C
1-6) alkyl, (C
3-7) cycloalkyl or (C
1-6) alkyl-(C
3-7) cycloalkyl, aryl, Het, (C
1-6Alkyl) aryl or (C
1-6Alkyl) Het, and R
112Be H, CN, (C
1-6) alkyl, (C
3-7) cycloalkyl or (C
1-6) alkyl-(C
3-7) cycloalkyl, aryl, Het, (C
1-6Alkyl) aryl, (C
1-6Alkyl) Het, COOR
115Or SO
2R
115, R wherein
115Be (C
1-6) alkyl, (C
3-7) cycloalkyl, (C
1-6) alkyl-(C
3-7) cycloalkyl, aryl, Het, (C
1-6Alkyl) aryl or (C
1-6Alkyl) Het, perhaps two R
111And R
112The nitrogen coupled by covalent linkage combines, formation 5,6 or the saturated heterocycle of 7-person, described alkyl, cycloalkyl, aryl, Het, (C
1-6Alkyl) aryl or (C
1-6Alkyl) Het, or heterocycle is all optional by R
160Replace;
F) NR
116COR
117, R wherein
116And R
117The H that respectively does for oneself, (C
1-6) alkyl, (C
3-7) cycloalkyl, (C
1-6) alkyl-(C
3-7) cycloalkyl, aryl, Het, (C
1-6Alkyl) aryl or (C
1-6Alkyl) Het, described (C
1-6) alkyl, (C
3-7) cycloalkyl, (C
1-6) alkyl-(C
3-7) cycloalkyl, aryl, Het, (C
1-6Alkyl) aryl or (C
1-6Alkyl) Het is all optional by R
160Replace;
G) NR
118CONR
119R
120, R wherein
118, R
119And R
120The H that respectively does for oneself, (C
1-6) alkyl, (C
3-7) cycloalkyl, (C
1-6) alkyl-(C
3-7) cycloalkyl, aryl, Het, (C
1- 6Alkyl) aryl or (C
1-6Alkyl) Het, perhaps R
118By covalent linkage and R
119In conjunction with and the nitrogen combination that is connected with it, form 5,6 or the saturated heterocycle of 7-person, perhaps R
119And R
120The nitrogen coupled by covalent linkage combines, formation 5,6 or the saturated heterocycle of 7-person, described alkyl, cycloalkyl, (C
1-6) alkyl-(C
3-7) cycloalkyl, aryl, Het, (C
1-6Alkyl) aryl or (C
1-6Alkyl) Het or heterocycle are all optional by R
160Replace;
H) NR
121COCOR
122, R wherein
121And R
122The H that respectively does for oneself, (C
1-6) alkyl, (C
3-7) cycloalkyl, (C
1-6) alkyl-(C
3-7) cycloalkyl, 6-or 10-person's aryl, Het, (C
1-6Alkyl) aryl or (C
1-6Alkyl) Het, described alkyl, cycloalkyl, alkyl-cycloalkyl, aryl, Het, (C
1-6Alkyl) aryl or (C
1-6Alkyl) Het is all optional by R
160Replace perhaps R
122Be OR
123Or N (R
124)
2, R wherein
123And each R
124Be H independently, (C
1-6Alkyl), (C
3-7) cycloalkyl, (C
1-6) alkyl-(C
3-7) cycloalkyl, aryl, Het, (C
1-6Alkyl) aryl or (C
1-6Alkyl) Het, perhaps R
124Be OH or O (C
1-6Alkyl), perhaps two R
124By covalent bonds together, formation 5,6 or the saturated heterocycle of 7-person, described alkyl, cycloalkyl, alkyl-cycloalkyl, aryl, Het, (C
1-6Alkyl) aryl or (C
1-6Alkyl) Het and heterocycle are all optional by R
160Replace;
I) COR
127, R wherein
127Be H, (C
1-6) alkyl, (C
3-7) cycloalkyl or (C
1-6) alkyl-(C
3-7) cycloalkyl, aryl, Het, (C
1-6Alkyl) aryl or (C
1-6Alkyl) Het, described alkyl, cycloalkyl, aryl, Het, (C
1-6Alkyl) aryl or (C
1-6Alkyl) Het is all optional by R
160Replace;
J) tetrazolium, COOR
128, R wherein
128Be H, (C
1-6) alkyl, (C
3-7) cycloalkyl, (C
1-6) alkyl-(C
3-7) cycloalkyl, aryl, Het, (C
1-6Alkyl) aryl or (C
1-6Alkyl) Het, described (C
1-6) alkyl, (C
3-7) cycloalkyl, (C
1-6) alkyl-(C
3-7) cycloalkyl, aryl, Het, (C
1-6Alkyl) aryl and (C
1-6Alkyl) Het is all optional by R
160Replace; And
K) CONR
129R
130, R wherein
129And R
130Be H independently, (C
1-6) alkyl, (C
3-7) cycloalkyl, (C
1-6) alkyl-(C
3-7) cycloalkyl, aryl, Het, (C
1-6Alkyl) aryl or (C
1-6Alkyl) Het, perhaps two R
129And R
130The nitrogen coupled by covalent linkage combines, formation 5,6 or the saturated heterocycle of 7-person, described alkyl, cycloalkyl, alkyl-cycloalkyl, aryl, Het, (C
1-6Alkyl) aryl, (C
1-6Alkyl) Het and heterocycle are all optional by R
160Replace;
R wherein
160Be defined as 1 or 2 substituting group, be selected from: tetrazolium, halogen, CN, C
1-6Alkyl, haloalkyl, COOR
161, SO
3H, SR
161, SO
2R
161, OR
161, N (R
162)
2, SO
2N (R
162)
2, NR
162COR
162Or CON (R
162)
2, R here
161And each R
162Be H independently, (C
1-6) alkyl, (C
3-7) cycloalkyl or (C
1-6) alkyl-(C
3-7) cycloalkyl; Perhaps two R
162The nitrogen coupled by covalent linkage combines, formation 5,6 or the saturated heterocycle of 7-person;
B is NR
3Or CR
3, condition is that one of A or B are CR
1Or CR
3, R wherein
3Be selected from: (C
1-6) alkyl, haloalkyl, (C
3-7) cycloalkyl, (C
5-7) cycloalkenyl group, (C
6-10) bicyclic alkyl, (C
6-10) bicycloenyl, 6-or 10-person's aryl, Het, (C
1-6) alkyl-aryl or (C
1-6) alkyl-Het,
Described alkyl, cycloalkyl, bicyclic alkyl, aryl, Het, alkyl-aryl and alkyl-Het be optional to be selected from following substituting group by 1～4 and to replace: halogen, perhaps
A) (C
1-6) alkyl, it is chosen wantonly and is replaced by following groups:
-OR
31Or SR
31, R wherein
31Be H, (C
1-6Alkyl), (C
3-7) cycloalkyl, (C
1-6) alkyl-(C
3-7) cycloalkyl, aryl, Het, (C
1-6) alkyl-aryl or (C
1-6) alkyl-Het; Perhaps
-N (R
32)
2, each R wherein
32Be H independently, (C
1-6) alkyl, (C
3-7) cycloalkyl, (C
1-6) alkyl-(C
3-7) cycloalkyl, aryl, Het, (C
1-6) alkyl-aryl or (C
1-6) alkyl-Het; Perhaps two R
32The nitrogen coupled by covalent linkage combines, formation 5,6 or the saturated heterocycle of 7-person;
B) OR
33, R wherein
33Be H, (C
1-6) alkyl, (C
3-7) cycloalkyl or (C
1-6) alkyl-(C
3-7) cycloalkyl, aryl, Het, (C
1-6) alkyl-aryl or (C
1-6) alkyl-Het;
C) SR
34, R wherein
34Be H, (C
1-6) alkyl, (C
3-7) cycloalkyl, perhaps (C
1-6) alkyl-(C
3-7) cycloalkyl, aryl, Het, (C
1-6) alkyl-aryl or (C
1-6) alkyl-Het; And
D) N (R
35)
2, each R wherein
35Be H independently, (C
1-6) alkyl, (C
3-7) cycloalkyl, (C
1-6) alkyl-(C
3-7) cycloalkyl, aryl, Het, (C
1-6) alkyl-aryl or (C
1-6) alkyl-Het; Perhaps two R
35The nitrogen coupled by covalent linkage combines, formation 5,6 or the saturated heterocycle of 7-person;
K is N or CR
4, R wherein
4Be H, halogen, (C
1-6) alkyl, haloalkyl, (C
3-7) cycloalkyl or (C
1-6) alkyl-(C
3-7) cycloalkyl; Perhaps R
4Be OR
41Or SR
41, COR
41Or NR
41COR
41, each R wherein
41Be H independently, (C
1-6) alkyl), (C
3-7) cycloalkyl or (C
1-6) alkyl-(C
3-7) cycloalkyl, perhaps R
4Be NR
42R
43, R wherein
42And R
43Be H independently, (C
1-6) alkyl, (C
3-7) cycloalkyl, (C
1-6) alkyl-(C
3-7) cycloalkyl, perhaps two R
42And R
43The nitrogen coupled by covalent linkage combines, formation 5,6 or the saturated heterocycle of 7-person;
L is N or CR
5, R wherein
5Definition and above-mentioned R
4Definition identical;
M is N or CR
7, R wherein
7Definition and above-mentioned R
4Definition identical;
Y
1Be O or S;
Z is OR
6, R wherein
6Be H, (C
1-6) alkyl, it is chosen wantonly and is replaced by following groups: halogen, hydroxyl, carboxyl, amino, C
1-6Alkoxyl group, C
1-6Alkoxy carbonyl, and C
1-6Alkylamino; Perhaps R
6Be C
1-6Alkylaryl, it is chosen wantonly and is replaced by following groups: halogen, cyano group, nitro, C
1-6Alkyl, C
1-6Haloalkyl, C
1-6Alkyloyl ,-(CH
2)
1-6-COOR
7,-(CH
2)
1-6-CONR
7R
8,-(CH
2)
1-6-NR
7R
8,-(CH
2)
1-6-NR
7COR
8,-(CH
2)
1-6-NHSO
2R
7,-(CH
2)
1-6-OR
7,-(CH
2)
1-6-SR
7,-(CH
2)
1-6-SO
2R
7, and-(CH
2)
1-6-SO
2NR
7R
8, each R wherein
7And each R
8Be H or C
1-6Alkyl,
Perhaps Z is NR
9R
10, each R wherein
9And R
10Be selected from: H, C
1-6Alkoxyl group, or C
1-6Alkyl, it is optional by halogen, hydroxyl, carboxyl, amino, C
1-6Alkoxyl group, C
1-6Alkoxy carbonyl, and C
1-6Alkylamino replaces;
Or its salt;
Condition is, when A is CR
1, R
1Be Me, R
2For pyridine or
, B is NR
3, R
3Be Me, K, L, M are CH, Y
1Be O, and Z is OR
6The time, R
6Be not H;
The condition of reaching is, when A is NR
1, R
1Be H, R
2Be phenyl, B is CR
3, R
3Be phenyl, K, L, M are CH, Y
1Be O, and Z is OR
6The time, R
6Be not H;
And condition is, when A is S, and R
2Be bromine, B is CR
3, R
3Be Me, K is CH, and L is CH, and M is CR
7, R
7Be H or Me, Y
1Be O, and Z is OR
6The time, R
6Be not H;
And condition is, when A is O, and R
2Be H, B is CR
3, R
3Be phenyl, K, L, M are CH, Y
1Be O, and Z is OR
6The time, R
6Be not H;
The condition of reaching is, when A is CR
1, R
1Be Me, R
2Be pyridine, B is NR
3, R
3Be Me, K, L, M are CH, Y
1Be O, and Z is OR
6The time, R
6Be not Me;
The condition of reaching is, when A is CR
1, R
1Be Me, R
2For
, B is NR
3, R
3Be Me, K, L, M are CH, Y
1Be O, and Z is OR
6The time, R
6Be not Et;
The condition of reaching is, when A is CR
1, R
1Be CH, R
2Be Me, B is NR
3, R
3Be Me, K, L, M are CH, Y
1Be O, and Z is OR
6The time R
6Be not Et;
The condition of reaching is, when A is CR
1, R
1Be Et, R
2Be Me, B is NR
3, R
3Be Me, K, L, M are CH, Y
1Be O, and Z is OR
6The time, R
6Be not CH
2CH
2N (Me)
2
And condition is, when A is CH, and R
2Be Me, B is NR
3, R
3For
, K is N, L is CR
5, R
5Be Me, M is CR
7, R
7Be OH, Y
1Be O, and Z is OR
6The time, R
6Be not Et;
The condition of reaching is, when A is NR
1, R
1Be Me, R
2Be Br, B is CR
3, R
3For
, K is N, L is CR
5, R
5Be Me, M is CR
7, R
7Be Br, Y
1Be O, and Z is OR
6The time, R
6Be not Me;
The condition of reaching is, when A is NR
1, R
1Be H, R
2Be Cl, B is CR
3, R
3Be Et, K is CH, and L is CH, and M is CH, Y
1Be O, Z is OR
6The time, R
6Be not Me;
The condition of reaching is, when A is NR
1, R
1Be H, R
2Be phenyl, B is CR
3, R
3Be phenyl, K is CH, and L is CH, and M is CR
7, R
7Be Me, Y
1Be O, Z is OR
6The time, R
6Be not Et;
The condition of reaching is, when A is NR
1, R
1Be H, R
2For
, B is CR
3, R
3For
, K is CH, and L is N, and M is CH, Y
1Be O, and Z is OR
6The time, R
6Be not Et;
And condition is, when A is S, and R
2Be Br, B is CR
3, R
3Be Me, K is CH, and L is CH, and M is CH, Y
1Be O, and Z is OR
6The time, R
6Be not Me;
The condition of reaching is, when A is NR
1, R
1Be H, R
2For
, B is NR
3, R
3Be cyclohexyl, K, L, M are CH, Y
1Be O, Z is OR
6The time, R
6Be not H.
As selection, in first aspect, the present invention also provides the compound shown in the following formula Ia:
In the formula:
A is O, S, NR
1, or CR
1
B is NR
3Or CR
3
R
1Be selected from: H, (C
1-6) alkyl, benzyl, (C
1-6Alkyl)-(C
6-10Aryl), (C
1-6Alkyl)-and 5-or 6-person have 1～4 heteroatomic heterocycle that is selected from O, N and S, and 5-or 6-person have 1～4 heteroatomic heterocycle that is selected from O, N and S,
Wherein said benzyl and described heteroatoms be optional to be selected from following substituting group by 1～4 and to replace:
COOH, COO (C
1-6Alkyl), halogen, and (C
1-6Alkyl);
R
2Be selected from: H, halogen, (C
1-6) alkyl, (C
3-7) cycloalkyl, phenyl, 5-or 6-person have 1～4 heteroatomic heterocycle that is selected from O, N and S, and pyridine-N-oxide reaches 9-or 10-person and has 1～4 heteroatomic assorted dicyclo that is selected from O, N and S,
Described phenyl, heterocycle and assorted dicyclo be optional to be selected from following substituting group by 1～4 and to replace: halogen, C (halogen)
3, (C
1-6) alkyl, OH, O (C
1-6Alkyl), NH
2, and N (C
1-6Alkyl)
2
R
3Be selected from: 5-, 6-or 7-person have 1～4 heteroatomic heterocycle that is selected from O, N and S, norcamphane, (C
3-7) cycloalkyl and (C
3-7) cycloalkyl-(C
1-6Alkyl);
M is N, CR
4, or COR
5, R wherein
4Be selected from: H, halogen, and (C
1-6Alkyl); And R
5Be selected from: H and (C
1-6Alkyl);
K and L are N or CH;
-----represent singly-bound or two key;
Y is O;
Z is OR
6Or NR
6R
6a, R wherein
6Be selected from: H, (C
1-6) alkyl, wherein said alkyl is optional to be selected from following substituting group by 1～4 and to replace: OH, COOH, COO (C
1-6) alkyl, (C
1-6) alkyl, described alkyl is optional to be selected from following substituting group by 1～4 and to replace: COOH, NHCO (C
1-6Alkyl), NH
2, NH (C
1-6Alkyl), reach N (C
1-6Alkyl)
2
Or its salt.
The third aspect the invention provides the compound or pharmaceutically acceptable salt thereof of formula I, as the inhibitor of the RNA RNA-dependent polymerase activity of the NS5B enzyme of HCV coding.
Fourth aspect the invention provides the compound or pharmaceutically acceptable salt thereof of formula I, the inhibitor that duplicates as HCV.
The 5th aspect the invention provides the method that treatment or prevention Mammals HCV infect, and comprises that the compound or pharmaceutically acceptable salt thereof with the formula I of significant quantity delivers medicine to Mammals.
The 6th aspect the invention provides the pharmaceutical composition that is used for the treatment of or prevents HCV to infect, and it comprises the compound or pharmaceutically acceptable salt thereof of formula I, and pharmaceutically useful carrier.
According to its specific embodiment, pharmaceutical composition of the present invention comprises extra immunomodulator.The example of extra immunomodulator include but not limited to α-, β-, δ-γ-, and ω-Interferon, rabbit.
According to alternative embodiment, pharmaceutical composition of the present invention can comprise antiviral agent extraly.The example of antiviral agent comprises virazole and amantadine.
According to another alternative embodiment, pharmaceutical composition of the present invention can comprise other HCV AG14361 extraly.
According to an alternative embodiment again, pharmaceutical composition of the present invention can comprise the inhibitor of other target in the HCV life cycle such as helicase, polysaccharase, metalloprotease or IRES extraly.
The 7th aspect, the purposes of the compound that the invention provides formula I in the medicine that preparation treatment HCV infects.
Eight aspect the invention provides the purposes of the compound of formula I as the HCV AG14361.
The 9th aspect the invention provides the method that a kind of treatment or prevention Mammals HCV infect, and comprises the compound or pharmaceutically acceptable salt thereof of the formula I of significant quantity and other anti-HCV medicament combination medicine-feeding in Mammals.
The tenth aspect the invention provides following formula (1a) or intermediate (1b):
Or
A in the formula, B, K, L, and M is as described herein, PG is H or carboxyl-protecting group.
The tenth on the one hand, the invention provides intermediate purposes in the formula compound (iii) below preparation of formula (Ia),
A in the formula, R
2, B, K, L, M, and PG is as described herein,
Comprise:
A) metal catalyst (as Pd, Ni, Ru, Cu), alkali and additive (as phosphine ligand, Cu salt, Li salt, ammonium salt CsF) exists down, in The suitable solvent, makes following intermediate (1a)
With R
2-X coupling, wherein R
1, R
3, K, L, M and PG as described herein, and X is (but being not limited to): Sn (C
1-6Alkyl)
3, Sn (aryl)
3, metal halide, B (OH)
2, and B (O (C
1-6) alkyl)
2, with preparation formula compound (iii).
In the tenth one side, the present invention alternatively provides intermediate (Ib) purposes in the formula compound (iii) below preparation:
A in the formula, R
2, B, K, L, M, and PG is as described herein,
Comprise:
B) metal catalyst (as Pd, Ni, Ru, Cu), alkali and additive (as phosphine ligand, Cu salt, Li salt, ammonium salt CsF) exists down, in The suitable solvent, makes following intermediate (1b)
With R
2-X ' coupling, X ' is a halogen in the formula, OSO
2(C
1-6Alkyl), OSO
2Ar, OSO
2CF
3Deng, and M is a metal, and as Li, Sn (C
1-6Alkyl)
3, Sn (aryl)
3, B (OH)
2, B (OC
1-6Alkyl)
2, metal halide is with preparation formula compound (iii).
Embodiment
Definition
Unless otherwise indicated, following definition is applicable to the present invention:
Term " (C used herein
1-3) alkyl ", " (C
1-4) alkyl " or " (C
1-6) alkyl ", no matter use separately still and use with other moiety combinations, all be meant aliphatics straight or branched alkyl, comprise nearly 3,4 and 6 carbon atoms respectively.This examples of groups comprises methyl, ethyl, propyl group, butyl, hexyl, 1-methylethyl, 1-methyl-propyl, 2-methyl-propyl, 1,1-dimethyl ethyl.
Term " (C used herein
2-6) alkenyl ", no matter use separately still and use with other moiety combinations, all be meant the unsaturated aliphatic straight chain group that comprises 2～6 carbon atoms.
Term " (C used herein
2-6) alkynyl " no matter use separately still and use with other moiety combinations, all be meant the group of the unsaturated aliphatic straight chain sp hydridization that comprises 2～6 carbon atoms.
Term " (C used herein
3-7) cycloalkyl ", no matter use separately still and use with other moiety combinations, all be meant the cycloalkyl that comprises 3～7 carbon atoms, comprise cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl and suberyl.
Term " (C used herein
5-7) cycloalkenyl group ", no matter use separately still and use with other moiety combinations, all be meant the undersaturated cyclic group that comprises 5～7 carbon atoms.
Term used herein " carboxyl-protecting group " is defined as the protecting group of using in the coupling, sees Greene for details, " Protective Groups in Organic Chemistry ", John Wiley ﹠amp; Sons, New York (1981) and " The Peptides:Analysis, Synthesis, Biology ", Vol.3, Academic Press, NewYork (1981), its disclosure is incorporated herein by reference.
The common protected one-tenth ester of α-carboxyl (CPG) of C-end residue, it can decompose and provides carboxylic acid.Spendable protecting group comprises: 1) alkyl ester; as methyl ester; trimethyl silyl ethyl ester and tertiary butyl ester; 2) aralkyl ester; benzyl ester as benzyl ester and replacement; perhaps 3) ester that can decompose by the alkaline purification or the gentle reduction of gentleness is as three chloro-ethyl esters and phenacyl ester.
No matter term used herein " aryl " or " 6-or 10-person's aryl " use separately still and use with other moiety combinations, all are meant the aryl that comprises 6 or 10 carbon atoms, for example phenyl or naphthyl.
Term used herein " heteroatoms " is meant O, S or N.
Term used herein " heterocycle ", no matter use separately still and use with other moiety combinations, be meant all that from 5-remove hydrogen atom in 6-or 7-person saturated or unsaturated (the containing aromaticity) heterocycle and the univalent perssad that obtains, this heterocycle comprises 1～4 heteroatoms that is selected from nitrogen, oxygen, sulphur.And " assorted two ring " used herein is meant defined above heterocyclic fused to one or more other rings, and it can be heterocycle or other ring.This heterocyclic example includes but not limited to tetramethyleneimine, tetrahydrofuran (THF), thiazolidine, pyrroles, thiophene, tonka bean camphor, glycolylurea, diazepine (diazepine), 1H-imidazoles ， isoxazole, thiazole, tetrazolium, piperidines, 1,4-diox, 4-morpholine, pyridine, pyridine-N-oxide, pyrimidine, thiazole also [4,5-b]-pyridine, quinoline, indoles, perhaps following heterocycle:
No matter term used herein " 9-or 10-person assorted two ring " or " heterocycle " use separately still and other moiety combinations use, and what all be meant above-mentioned definition is heterocyclic fused to one or more other rings, and it can be heterocycle or other ring.This heterocyclic example includes but not limited to also [4,5-b]-pyridine of thiazole, quinoline, and indoles, perhaps following heterocycle:
Term used herein " Het " is defined as has 1～4 heteroatomic 5-or 6-element heterocycle that is selected from O, N and S, perhaps may have 1～5 heteroatomic 9-or assorted two ring of 10-person that are selected from O, N and S under the situation.
Term used herein " halogen " is meant halogen atom, comprises fluorine, chlorine, bromine, iodine.
Term used herein " haloalkyl " is meant that wherein each hydrogen atom is successively by the displaced abovementioned alkyl of halogen atom, for example CH
2Br or CF
3
Term used herein " metal halide " be meant use in the cross-coupling reaction of metal catalytic with any metal of halogen atom bonded.The example of this metal halide includes but not limited to-MgCl ,-CuCl ,-ZnCl etc.
Term used herein " OH " is meant hydroxyl.Those skilled in the art is well-known to be, hydroxyl can be replaced by the functional group of equivalence.The example of used this equivalent functional group includes but not limited to ether among the present invention, sulfydryl, and primary amine, secondary amine or tertiary amine.
Term used herein " SH " is meant sulfydryl.Within the scope of the invention, " SH " or " SR " group no matter occurs, be meant that all this group can also be by any other suitable oxidation state such as SOR, SO
2R or SO
3R replaces.
Group such as C that term " replacement " is above with having a part
1-6Alkyl-aryl or C
1-6During alkyl-Het logotype, be meant in two portions and all can use this replacement, that is to say that alkyl and aryl or Het all can be replaced by defined substituting group.
Term used herein " COOH " is meant hydroxy-acid group.Those skilled in the art is well-known to be, hydroxy-acid group can be replaced by the functional group of equivalence.The example of this equivalent functional group of the present invention includes but not limited to ester, acid amides, boric acid (boronic acid) or tetrazolium.
Term used herein " functional group of equal value " is meant the derivative of element or its replacement, and it can or become other similar element of key character to replace by electronics, hydridization.
Term used herein " metal catalyst " be meant use in the cross-coupling reaction with leavings group bonded metal such as palladium (0) or palladium (2).The example of this palladium catalyst includes but not limited to Pd (Ph
3)
4, Pd/C, Pd (OAc)
2, PdCl
2Deng.Alternative metal that can the catalysis cross-coupling reaction includes but not limited to Ni (acac)
2, Ni (OAc)
2Or NiCl
2
Term used herein " derivative " is meant " can detected marker ", " affinity tag " or " photoreactive groups ".Term " can " be meant any group that can be connected on polysaccharase or the compound of the present invention by detected marker, make when this compound combines with the target polymerization enzyme, this marker allows to discern described compound directly or indirectly, and then this compound of detection, measurement and quantification.The example of this " marker " includes but not limited to fluorescent marker, chemiluminescent labels, colorimetric marker, enzyme labelling thing, radio isotope and affinity tag such as vitamin H.This marker is connected on compound or the polysaccharase by well-known method.
Term " affinity tag " is meant a kind of part (it links to each other with polysaccharase or compound of the present invention), and it can be used for extracting the entity that this part connected to acceptor strong affine from solution.The example of this part comprises the vitamin H or derivatives thereof, Histidine polypeptide, poly arginine, straight-chain polysaccharide part or the appointment epitope that can be discerned by antibody specific.This affinity tag is connected on compound or the polysaccharase by well-known method.
Term " photoreactive groups " is meant such group, and it can change into the group with reactive behavior, for example free radical from inertia group by photoactivation.This examples of groups includes but not limited to benzophenone, trinitride etc.
Term used herein " pharmaceutical salts " comprises that those come from the derivative of pharmaceutically acceptable alkali, and is nontoxic.The example of suitable alkali comprises choline, thanomin and quadrol.Na
+, K
+, and Ca
++Salt be also included within the scope of the present invention (can also be with reference to Pharmaceutical Salts, Birge, S.M.et al., J.Pharm.Sci., (1977),
66, 1-19, the document is incorporated herein by reference).
Preferred embodiment
A：
Preferred compound of the present invention has as shown in the formula the structure shown in (II):
In the formula, preferred A is O, S or NR
1
Preferred A is NR
1
Preferred compound of the present invention has as shown in the formula the structure shown in (III):
In the formula, preferred B is NR
3
For formula (II) and compound (III), preferred M, K and L are CH or N.More preferably M, K and L are CH.
Compound more preferably of the present invention has following structure shown in various:
R
1
：
Preferred R
1Be selected from: H or (C
1-6) alkyl.More preferably R
1Be H, CH
3, sec.-propyl, or isobutyl-.
Also preferred R
1Be H or CH
3R most preferably
1Be CH
3
R
2
：
Preferred R
2Be selected from: H, halogen, (C
2-6) alkenyl, (C
5-7) cycloalkenyl group, 6-or 10-person's aryl or Het; (C wherein
2-6) alkenyl, (C
5-7) cycloalkenyl group, aryl or Het are optional by R
20Replace R wherein
20Be defined as:
-1～4 substituting groups, it is selected from: halogen, NO
2, cyano group, azido-,
C (=NH) NH
2, C (=NH) NH (C
1-6) alkyl or C (=NH) NHCO (C
1-6) alkyl; Perhaps-1～4 substituting group, it is selected from:
A) (C
1-6) alkyl or haloalkyl, (C
3-7) cycloalkyl, (C
2-6) alkenyl, (C
2-8) alkynyl, (C
1-6) alkyl-(C
3-7) cycloalkyl, all these substituting groups are all randomly by R
150Replace;
B) OR
104, R wherein
104Be H, (C
1-6Alkyl), (C
3-7) cycloalkyl, or (C
1-6) alkyl-(C
3-7) cycloalkyl, aryl, Het, (C
1-6Alkyl) aryl or (C
1-6Alkyl) Het, described alkyl, cycloalkyl, aryl, Het, (C
1-6Alkyl) aryl or (C
1-6Alkyl) Het is all randomly by R
150Replace;
C) OCOR
105, R wherein
105Be (C
1-6) alkyl, (C
3-7) cycloalkyl, (C
1-6) alkyl-(C
3-7) cycloalkyl, Het, (C
1-6Alkyl) aryl or (C
1-6Alkyl) Het, described alkyl, cycloalkyl, aryl, Het, (C
1-6Alkyl) aryl or (C
1-6Alkyl) Het is all randomly by R
150Replace;
D) SR
108, SO
2N (R
108)
2Or SO
2N (R
108) C (O) R
108, each R wherein
108Be H independently, (C
1-6) alkyl, (C
3-7) cycloalkyl or (C
1-6) alkyl-(C
3-7) cycloalkyl, aryl, Het, (C
1-6Alkyl) aryl or (C
1-6Alkyl) Het or two R
108The nitrogen coupled by covalent linkage combines, formation 5,6 or the saturated heterocycle of 7-person, described alkyl, cycloalkyl, aryl, Het, (C
1-6Alkyl) aryl or (C
1-6Alkyl) Het or heterocycle are all randomly by R
150Replace;
E) NR
111R
112, R wherein
111Be H, (C
1-6) alkyl, (C
3-7) cycloalkyl or (C
1-6) alkyl-(C
3-7) cycloalkyl, aryl, Het, (C
1-6Alkyl) aryl or (C
1-6Alkyl) Het, and R
112Be H, CN, (C
1-6) alkyl, (C
3-7) cycloalkyl or (C
1-6) alkyl-(C
3-7) cycloalkyl, aryl, Het, (C
1-6Alkyl) aryl, (C
1-6Alkyl) Het, COOR
115Or SO
2R
115, R wherein
115Be (C
1-6) alkyl, (C
3-7) cycloalkyl, or (C
1-6) alkyl-(C
3-7) cycloalkyl, aryl, Het, (C
1-6Alkyl) aryl or (C
1-6Alkyl) Het, perhaps two R
111And R
112The nitrogen coupled by covalent linkage combines, formation 5,6 or the saturated heterocycle of 7-person, described alkyl, cycloalkyl, aryl, Het, (C
1-6Alkyl) aryl or (C
1-6Alkyl) Het, or heterocycle is all randomly by R
150Replace;
F) NR
116COR
117, R wherein
116And R
117The H that respectively does for oneself, (C
1-6) alkyl, (C
3-7) cycloalkyl, (C
1-6) alkyl-(C
3-7) cycloalkyl, aryl, Het, (C
1-6Alkyl) aryl or (C
1-6Alkyl) Het, described (C
1-6) alkyl, (C
3-7) cycloalkyl, (C
1-6) alkyl-(C
3-7) cycloalkyl, aryl, Het, (C
1-6Alkyl) aryl or (C
1-6Alkyl) Het is all randomly by R
150Replace;
G) NR
118CONR
119R
120, R wherein
118, R
119And R
120The H that respectively does for oneself, (C
1-6) alkyl, (C
3-7) cycloalkyl, (C
1-6) alkyl-(C
3-7) cycloalkyl, aryl, Het, (C
1-6Alkyl) aryl or (C
1-6Alkyl) Het, perhaps R
118By covalent linkage and R
119And the nitrogen that is connected combines formation 5,6 or the saturated heterocycle of 7-person;
Perhaps R
119And R
120The nitrogen coupled by covalent linkage combines, formation 5,6 or the saturated heterocycle of 7-person;
Described alkyl, cycloalkyl, (C
1-6) alkyl-(C
3-7) cycloalkyl, aryl, Het, (C
1-6Alkyl) aryl or (C
1-6Alkyl) Het or heterocycle are all randomly by R
150Replace;
H) NR
121COCOR
122, R wherein
121And R
122The H that respectively does for oneself, (C
1-6) alkyl, (C
3-7) cycloalkyl, (C
1-6) alkyl-(C
3-7) cycloalkyl, 6-or 10-person's aryl, Het, (C
1-6Alkyl) aryl or (C
1-6Alkyl) Het, described alkyl, cycloalkyl, alkyl-cycloalkyl, aryl, Het, (C
1-6Alkyl) aryl or (C
1-6Alkyl) Het is all randomly by R
150Replace;
Perhaps R
122Be OR
123Or N (R
124)
2, R wherein
123And each R
124Be H independently, (C
1-6Alkyl), (C
3-7) cycloalkyl, (C
1-6) alkyl-(C
3-7) cycloalkyl, aryl, Het, (C
1-6Alkyl) aryl or (C
1-6Alkyl) Het, perhaps R
124Be OH or O (C
1-6Alkyl), perhaps two R
124By covalent bonds together, formation 5,6 or the saturated heterocycle of 7-person, described alkyl, cycloalkyl, alkyl-cycloalkyl, aryl, Het, (C
1-6Alkyl) aryl or (C
1-6Alkyl) Het and heterocycle are all randomly by R
150Replace;
I) COR
127, R wherein
127Be H, (C
1-6) alkyl, (C
3-7) cycloalkyl or (C
1-6) alkyl-(C
3-7) cycloalkyl, aryl, Het, (C
1-6Alkyl) aryl or (C
1-6Alkyl) Het, described alkyl, cycloalkyl, aryl, Het, (C
1-6Alkyl) aryl or (C
1-6Alkyl) Het is all randomly by R
150Replace;
J) COOR
128, R wherein
128Be H, (C
1-6) alkyl, (C
3-7) cycloalkyl, (C
1-6) alkyl-(C
3-7) cycloalkyl, aryl, Het, (C
1-6Alkyl) aryl or (C
1-6Alkyl) Het, described (C
1-6) alkyl, (C
3-7) cycloalkyl, or (C
1-6) alkyl-(C
3-7) cycloalkyl, aryl, Het, (C
1-6Alkyl) aryl and (C
1-6Alkyl) Het is all randomly by R
150Replace;
K) CONR
129R
130, R wherein
129And R
130Be H independently, (C
1-6) alkyl, (C
3-7) cycloalkyl, (C
1-6) alkyl-(C
3-7) cycloalkyl, aryl, Het, (C
1-6Alkyl) aryl or (C
1-6Alkyl) Het, perhaps two R
129And R
130The nitrogen coupled by covalent linkage combines, formation 5,6 or the saturated heterocycle of 7-person, described alkyl, cycloalkyl, alkyl-cycloalkyl, aryl, Het, (C
1-6Alkyl) aryl, (C
1-6Alkyl) Het and heterocycle are all randomly by R
150Replace;
L) aryl, Het, (C
1-6Alkyl) aryl or (C
1-6Alkyl) Het, all these groups are all randomly by R
150Replace;
R wherein
150Be preferably:
-1～3 substituting groups, it is selected from: halogen, NO
2, cyano group or azido-; Perhaps
-1～3 substituting groups, it is selected from:
A) (C
1-6) alkyl or haloalkyl, (C
3-7) cycloalkyl, (C
2-6) alkenyl, (C
2-8) alkynyl, (C
1-6) alkyl-(C
3-7) cycloalkyl, all these groups are all randomly by R
160Replace;
B) OR
104, R wherein
104Be H, (C
1-6Alkyl) or (C
3-7) cycloalkyl, described alkyl or cycloalkyl is all randomly by R
160Replace;
D) SR
108, SO
2N (R
108)
2Or SO
2N (R
108) C (O) R
108, each R wherein
108Be H independently, (C
1-6) alkyl, (C
3-7) cycloalkyl or (C
1-6) alkyl-(C
3-7) cycloalkyl, aryl, Het, perhaps two R
108The nitrogen coupled by covalent linkage combines, formation 5,6 or the saturated heterocycle of 7-person, and described alkyl, cycloalkyl, aryl, Het and heterocycle are all optional by R
160Replace;
E) NR
111R
112, R wherein
111Be H, (C
1-6) alkyl, or (C
3-7) cycloalkyl, and R
112Be H, (C
1-6) alkyl or (C
3-7) cycloalkyl, COOR
115Or SO
2R
115, R wherein
115Be (C
1-6) alkyl or (C
3-7) cycloalkyl, perhaps two R
111And R
112The nitrogen coupled by covalent linkage combines, and formation 5,6 or the saturated heterocycle of 7-person, described alkyl, cycloalkyl and heterocycle are all optional by R
160Replace;
F) NR
116COR
117, R wherein
116And R
117The H that respectively does for oneself, (C
1-6) alkyl or (C
3-7) cycloalkyl, described (C
1-6) alkyl and (C
3-7) cycloalkyl is all optional by R
160Replace;
G) NR
118CONR
119R
120, R wherein
118, R
119And R
120The H that respectively does for oneself, (C
1-6) alkyl or (C
3-7) cycloalkyl, perhaps R
118By covalent linkage and R
119In conjunction with also coupled nitrogen combination, formation 5,6 or the saturated heterocycle of 7-person;
Perhaps R
119And R
120The nitrogen coupled by covalent linkage combines, formation 5,6 or the saturated heterocycle of 7-person;
Described alkyl, cycloalkyl, and heterocycle is all optional by R
160Replace;
H) NR
121COCOR
122, R wherein
121Be H, (C
1-6) alkyl or (C
3-7) cycloalkyl, described alkyl and cycloalkyl are all optional by R
160Replace;
Perhaps R
122Be OR
123Or N (R
124)
2, R wherein
123And each R
124Be H independently, (C
1-6Alkyl) or (C
3-7) cycloalkyl, perhaps two R
124By covalent bonds together, formation 5,6 or the saturated heterocycle of 7-person, described alkyl, cycloalkyl and heterocycle are all optional by R
160Replace;
I) COR
127, R wherein
127Be H, (C
1-6) alkyl or (C
3-7) cycloalkyl, described alkyl and cycloalkyl are all optional by R
160Replace;
J) COOR
128, R wherein
128Be H, (C
1-6) alkyl or (C
3-7) cycloalkyl, described (C
1-6) alkyl and (C
3-7) cycloalkyl is all optional by R
160Replace; And
K) CONR
129R
130, R wherein
129And R
130Be H independently, (C
1-6) alkyl or (C
3-7) cycloalkyl, perhaps two R
129And R
130The nitrogen coupled by covalent linkage combines, and formation 5,6 or the saturated heterocycle of 7-person, described alkyl, cycloalkyl and heterocycle are all optional by R
160Replace;
R wherein
160Be defined as 1 or 2 substituting groups, it is selected from:
Halogen, CN, C
1-6Alkyl, haloalkyl, COOR
161, OR
161, N (R
162)
2, SO
2N (R
162)
2, NR
162COR
162Or CON (R
162)
2, R wherein
161And each R
162Be H independently, (C
1-6) alkyl, (C
3-7) cycloalkyl or (C
1-6) alkyl-(C
3-7) cycloalkyl; Perhaps two R
162The nitrogen coupled by covalent linkage combines, formation 5,6 or the saturated heterocycle of 7-person.
More preferably R
2Be selected from: aryl or Het, each all randomly is selected from following substituting group list and is replaced or two replacements: halogen, haloalkyl, N
3, perhaps
A) (C
1-6) alkyl, optional by OH, O (C
1-6) alkyl or SO
2(C
1-6Alkyl) replaces;
B) (C
1-6) alkoxyl group;
E) NR
111R
112, R wherein
111And R
112Be H independently, (C
1-6) alkyl, (C
3-7) cycloalkyl, perhaps R
112Be 6-or 10-person's aryl, Het, (C
1-6) alkyl-aryl or (C
1-6) alkyl-Het; Perhaps two R
111And R
112The nitrogen coupled by covalent linkage combines, and forms nitrogenous heterocycle, described alkyl, and cycloalkyl, aryl, Het, each among alkyl-aryl or the alkyl-Het is all randomly replaced by following groups: halogen, perhaps
-OR
161Or N (R
162)
2, R wherein
161And each R
162Be H independently, (C
1-6) alkyl, perhaps two R
162The nitrogen coupled by covalent linkage combines, and forms nitrogenous heterocycle;
F) NHCOR
117, R wherein
117Be (C
1-6) alkyl, O (C
1-6) alkyl or O (C
3-7) cycloalkyl;
I) CO-aryl; And
K) CONH
2, CONH (C
1-6Alkyl), CON (C
1-6Alkyl)
2, CONH-aryl, or CONHC
1-6Alkyl-aryl.
Also preferred R
2Be aryl or Het, each all randomly is selected from following substituting group list and is replaced or two replacements: halogen, and haloalkyl, perhaps
A) (C
1-6) alkyl, it is optional by OH, O (C
1-6) alkyl or SO
2(C
1-6Alkyl) replaces;
B) (C
1-6) alkoxyl group; And
E) NR
111R
112, R wherein
111And R
112Be H independently, (C
1-6) alkyl, (C
3-7) cycloalkyl, perhaps R
112Be 6-or 10-person's aryl, Het, (C
1-6) alkyl-aryl or (C
1-6) alkyl-Het; Perhaps two R
111And R
112The nitrogen coupled by covalent linkage combines, and forms nitrogenous heterocycle, described alkyl, and cycloalkyl, aryl, Het, each among alkyl-aryl or the alkyl-Het is all randomly replaced by following groups: halogen, perhaps
-OR
161Or N (R
162)
2, R wherein
161And each R
162Be H independently, (C
1-6) alkyl, perhaps two R
162The nitrogen coupled by covalent linkage combines, and forms nitrogenous heterocycle.
Even more preferably R
2For phenyl or be selected from following heterocycle:
Even more preferably R
2Be selected from:
H，Br，CH＝CH
2，
More preferably R also
2Be selected from:
R most preferably
2Be selected from:
R
3
：
Preferred R
3Be selected from: (C
3-7) cycloalkyl, (C
3-7) cycloalkenyl group, (C
6-10) bicyclic alkyl, (C
6-10) bicycloenyl, 6-or 10-person's aryl, perhaps Het.More preferably R
3Be (C
3-7) cycloalkyl.R most preferably
3Be cyclopentyl or cyclohexyl.
Y：
Preferred Y
1Be O.
Z：
Preferred Z is OR
6, R wherein
6Be H, (C
1-6) alkyl, it is chosen wantonly and is replaced by following groups: halogen, hydroxyl, carboxyl, amino, C
1-6Alkoxyl group, C
1-6Alkoxy carbonyl, and C
1-6Alkylamino; Perhaps R
6Be C
1-6Alkylaryl, it is chosen wantonly and is replaced by following groups: halogen, cyano group, nitro, C
1-6Alkyl, C
1-6Haloalkyl, C
1-6Alkyloyl ,-(CH
2)
1-6-COOR
7,-(CH
2)
1-6-CONR
7R
8,-(CH
2)
1-6-NR
7R
8,-(CH
2)
1-6-NR
7COR
8,-(CH
2)
1-6-NHSO
2R
7,-(CH
2)
1-6-OR
7,-(CH
2)
1-6-SR
7,-(CH
2)
1-6-SO
2R
7, and-(CH
2)
1-6-SO
2NR
7R
8, each R wherein
7With each R
8Be H or C
1-6Alkyl,
Perhaps Z is NR
9R
10, R wherein
9And R
10In each be selected from: H, C
1-6Alkoxyl group, or C
1-6Alkyl, it is randomly by halogen, hydroxyl, carboxyl, amino, C
1-6Alkoxyl group, C
1-6Alkoxy carbonyl, and C
1-6Alkylamino replaces;
More preferably Z is OH or O (C
1-6Alkyl), perhaps Z is NR
9R
10, R wherein
9Be preferably H, and R
10Be preferably H or C
1-6Alkyl.
Most preferably Z is OH.
Specific embodiments
The present invention includes the compound of all formula I, as shown in table 1 and 2.
Polymerase activity
The RNA RNA-dependent polysaccharase of formula (I) compound by HCV suppresses RNA synthetic ability, can by any can measure R NA RNA-dependent polymerase activity test confirm.Suitable test is described among the embodiment.
The specificity of RNA RNA-dependent polymerase activity
In order to confirm that compound of the present invention plays a role by suppressing the HCV polysaccharase specifically, can in the dna dependent rna polymerase assay, check the inhibition activity of compound.
When one of compound or pharmaceutically acceptable salt thereof of formula (I) during as antiviral agent, it can per os, part or whole body deliver medicine to Mammals (as people, rabbit or mouse) in solvent, described solvent comprises one or more pharmaceutically acceptable carriers, its ratio is according to the solubleness and the chemical property of compound, and the biology convention of selected route of administration and standard is determined.
For oral administration, compound or pharmaceutically acceptable salt thereof can be mixed with form such as the capsule or the tablet of unitary dose, each self-contained predetermined amount, and the activeconstituents that is generally 25～500mg is in pharmaceutically acceptable carrier.
For topical, compound can be formulated in the pharmaceutically acceptable solvent, and it comprises 0.1～5%, preferred 0.5～5% active agents.This formulation can be solution, emulsion or lotion.For parenteral admin, the compound of formula (I) can with the form of the composition of pharmaceutically acceptable solvent or carrier, by intravenous injection, subcutaneous injection or administered intramuscular.For drug administration by injection, preferably use the solution of compound in aseptic aqueous vehicle, it also can comprise other solute such as buffer reagent or sanitas, and the pharmacy acceptable salt of q.s or glucose, so that the solution isoosmotic pressure.
The solvent or the carrier that are suitable for above-mentioned formulation can make reference to the text-book, for example " Remington ' s The Scienceand Practice of Pharmacy ", 19th ed., Mack Publishing Company, Easton, Penn., 1995, perhaps " Pharmaceutical Dosage Forms And Drugs Delivery Systems ", 6thed., H.C.Ansel et al., Eds., Williams ﹠amp; Wilkins, Baltimore, Maryland, 1995.
The dosage of compound can be along with form of medication and selected concrete active agents and is changed.And it can also change according to concrete treatment host.Usually, treatment begins with little increment, progressively reaches the best effect under this situation.Usually, most preferably the compound of formula I is with such concentration level administration, promptly can provide antiviral effect but do not cause the concentration level of any injury or harmful side effect.
For oral administration, compound or pharmaceutically acceptable salt thereof is with per kg body weight per day 10～200mg, the amount administration of preferred 25～150mg.
For the whole body administration, the compound of formula (I) is with the dosed administration of per kg body weight per day 10～150mg, although aforesaid change can occur.In order to obtain effect, most preferably the dosage level of Shi Yonging is per kg body weight per day 10～100mg.
When composition of the present invention comprised the combination of the compound of formula I and one or more healing potions or prevention medicament, the dosage of compound and other medicament all should be the about 10～100% of bio-occlusion pharmaceutical quantities in the single therapy mode, and more preferably from about 10～80%.
When these compound or pharmaceutically acceptable salt thereofs are prepared with pharmaceutically acceptable carrier, resulting composition can vivo medicine-feeding in Mammals (as the people), to suppress HCV polysaccharase or treatment or prevention HCV virus infection.This treatment also can utilize the combination of compound of the present invention and following medicament to realize that described medicament includes but not limited to: immunomodulator, as α-, β-, or gamma-interferon; Other antiviral agent, as virazole, amantadine; Other HCV NS5B AG14361; The inhibitor of other target in the HCV life cycle, this target includes but not limited to helicase, NS2/3 proteolytic enzyme, NS3 proteolytic enzyme, or internal ribosome entry site (IRES); Perhaps their combination.Other medicament can with compound of the present invention, make single formulation.As selection, the part that these other medicaments can be used as multiple formulation is individually dosed.
Method is with synthetic
Indole derivatives of the present invention or analogue can utilize known literature method to prepare by known monocycle aromatic compound, referring to J.W.Ellingboe et al. (Tet.Lett.1997,38,7963) and S.Cacchi et al. (Tet.Lett.1992,33,3915).Below shown in scheme 1, R wherein
1, R
2, R
3, R
6, K, L, and M is as described herein, illustrates the compound that can how to adopt these methods to synthesize formula I of the present invention.
Scheme 1
Shown in the scheme 1 in the enforcement of approach, the 3-trifluoroacetamido-4-iodobenzoic acid I (i) that makes due care and alkynes I are (ii) at metal catalyst (for example metallic palladium complex compound such as PdCl
2(PPh
3)
2, Pd
2Dba
3, Pd (PPh
3)
4Deng), alkali (Et
3N, DIEA etc., perhaps alkaline, inorganic salts comprises metal carbonate, fluorochemical and phosphoric acid salt) exist down, and optional other phosphine ligand (triaryl or heteroaryl phosphine, dppe, dppf, dppp etc.) is left and is reacted.The suitable solvent of this reaction comprises DMF ， diox, THF, and DME, toluene, MeCN, DMA etc., suitable temperature is 20～170 ℃, as selection, can also react by heating each component together without solvent.Alternatively be; this cross-coupling reaction can also carry out at the 3-amino-4-iodobenzoic acid salt of due care; and described amino can carry out trifluoroacetylation by the subsequent step described in the J.W.Ellingboe et al. (Tet.Lett.1997,38,7963).
Above-mentioned diaryl alkynes I (iii) with the enol triflate in the reaction that is similar under the above-mentioned cross-coupling reaction condition, after the two keys of hydrogenation, provide indole derivatives I (iv).The enol triflate is known, and can (for example prepare by corresponding ketone according to following known literature method, the tetrahydrobenzene triflate can be by pimelinketone, and trifluoromethanesulfanhydride anhydride and sterically hindered organic bases are as 2, and 6-di-t-butyl-4-picoline prepares).Be present in R at first
1In two keys can utilize hydrogen or hydrogen donor (ammonium formiate, formic acid etc.), in the presence of metal catalyst (preferred Pd), in appropriate solvent (lower alkyl alcohol, THF etc.), carry out hydrogenation.
At last, after the ester protecting group of hydrolysis I in (iv), with gained 6-carboxyl indole derivative I (v) by with formula H
2N-R
6Suitable amine coupling and transform the compound of accepted way of doing sth I.6-indole-carboxylic acid and amine H
2N-R
6Condensation can utilize the amido linkage of standard to generate reagent such as TBTU, HATU, BOP, BroP, EDAC, DCC, isobutyl chlorocarbonate wait and realize, perhaps by activated carboxyl, promptly with the amine condensation before carboxyl changed into corresponding acyl chlorides realize.After this step, remove the protecting group of any remnants, promptly obtain the compound of formula I.
As selection, the compound of formula I can also be according to the literature method of revising, form by the indole nucleus that is pre-existing in is refining, (Tetrahedron 1996 referring to P.Gharagozloo et al. for the example of described literature method, 52,10185) or K.Freter (J.Org.Chem.1975,40,2525).This method is illustrated in the scheme 2.
Scheme 2
In the enforcement of approach shown in the scheme 2, adopt commercial available 6-indole-carboxylic acid 2 (i) as raw material, it also can be according to people's such as S.Kamiya method (Chem.Pharm.Bull.1995,43,1692) preparation.Indoles 2 (i) (ii) reacts under alkalescence or acid alcohol aldehyde-type reaction conditions with ketone 2.The suitable reaction condition that influences this condensation comprises highly basic such as alkali metal hydroxide, alkoxide and hydride in solvent, and the example of described solvent comprises lower alkyl alcohol (MeOH, EtOH, tertBuOH etc.), THF ， diox, DMF, DMSO, DMA etc., temperature of reaction is-20～120 ℃.As selection, this condensation can utilize organic acid or mineral acid or organic acid and mineral acid to carry out under acidic conditions.Suitable condition comprises the mixture of AcOH and phosphate aqueous solution, and 15～120 ℃ temperature.
Utilize after the form protection hydroxy-acid group of currently known methods with ester (being generally lower alkyl esters), can use R as required
3Make the indole nitrogen alkylation.The reaction conditions that makes the azanylization of indole derivatives is that those skilled in the art is well-known, and be included in and use highly basic such as alkalimetal hydride under-78～140 ℃ the temperature, oxyhydroxide, amides, alkoxide and metal alkylide in The suitable solvent (as THF diox, DME, DMF, MeCN, DMSO, alcohol etc.) in.Use the R of close electric form
3The indoles negatively charged ion is carried out alkylation.This close isoelectric substance comprises iodide, bromide, muriate and sulphonate (methanesulfonates, tosylate, brosylate or triflate).
Halogenation (generally being bromination, also can be iodate) indoles 2 2-position (iv) obtains 2 (v).Suitable halogenating agent comprises for example elemental bromine, N-bromosuccinimide, pyridine tribromide, two bromo glycolylurea and corresponding iodo derivative.The solvent that is suitable for this reaction is inertia to the halogenating agent with reactive behavior, and comprises for example hydrocarbon, chlorinated hydrocarbon (DCM, CCl
4, CHCl
3), ether (THF, DME ， diox), acetate, ethyl acetate, IPA, and the mixture of these solvents.Temperature of reaction is-40～100 ℃.See L.Chu (Tet.Lett.1997,38,3871) for carrying out the method that the indoles bromination shown in the scheme 2 selects.
2-bromo indole derivative 2 (v) can by with aryl or heteroaryl boric acid (boronic acid), the key intermediate I that the cross-coupling reaction of boric acid ester or trialkyltin alkane derivatives and directly change into fully replaces is (v).These boron or tin organo-metallic material can obtain from commercial source, also can be by the literature method preparation of standard.Can be undertaken by the variant of any Suzuki cross-coupling reaction of being reported in the document with the cross-coupling reaction of organoboron reagent.This generally includes and uses transition-metal catalyst (normally Pd °), triaryl or three heteroaryl phosphine ligands, additive such as butter (for example LiCl), and alkali (the normally carbonate of mineral alkali such as sodium or potassium or the phosphatic aqueous solution).This reacts generally at alcoholic solvent (EtOH), DME, and toluene among the THF etc., carries out under 25～140 ℃ temperature.
Can be undertaken by any variant of the Stille cross-coupling reaction reported in the document with the cross-coupling of tin reagent.This generally includes and uses transition-metal catalyst (being generally Pd °), triaryl or three heteroaryl phosphine ligands, and additive such as butter (for example LiCl) or iodide (for example CuI).The solvent that is suitable for this reaction comprises toluene, DMF, and THF, DME etc., temperature of reaction is 25～140 ℃.As described in the scheme 1, intermediate compound I (is v) transformed the compound of accepted way of doing sth I then.
As selection, 2-bromo indole intermediate 2 (v) can change into organotin (or organic zinc) material by trans-metal, and be used for Stille-type cross-coupling reaction under these conditions.In this case, utilize the halogenide (muriate, bromide, iodide) of aromatic series or heteroaromatic or triflate to introduce R
2(v) (conversion vi) is to utilize alkyl lithium reagents (for example n-BuLi or tert-BuLi) or utilize the lithium metal to 2-bromo indole derivative 2, is undertaken by the halogen-metal exchange of initial low temperature (being generally-78～-30 ℃) to corresponding organotin material 2.Then, the 2-lithium of transient state for indoles by trialkyltin halogenide (n-Bu for example
3SnCl or Me
3SnCl) catch.As selection, lithium can be chlorinated zinc for the indoles intermediate and catch, form corresponding organic zinc hydrochlorate, this organic zinc hydrochlorate can described aromatic series and the halogenide or the triflate of heteroaromatic carry out transition metal-catalyzed cross-coupling reaction, for example referring to M.Rowley (J.Med.Chem.2001,44,1603).
The present invention comprises that also carboxyl is positioned at the compound of the formula I of indoles system 5-position.The synthetic literature method of this compound based on correction, and be illustrated in the scheme 3.
Scheme 3
In the enforcement of synthesis path shown in the scheme 3,4-acetylaminohydroxyphenylarsonic acid 3-iodobenzoic acid ethyl ester 3 (i) and alkynes 3 (ii) carry out the cross-coupling of metal catalytic, obtain 2,3-is dibasic-and 5-indoxyl carboxylic acid ester 3 is (iii), the method that is adopted is seen A.Bedeschi et al. (Tet.Lett.1997,38,2307).Then, with the electric R of parent
1Group (halogenide, sulphonate), under the effect of alkali such as alkali-metal oxyhydroxide, fluorochemical, hydride, amides, lithium alkylide, phosphoric acid salt etc., alkylated indoles derivative 3 (iii) obtains 3 (iv) on nitrogen.This alkylating suitable solvent comprises DMF, DMA, DMSO, MeCN, THF ， diox, DME etc.After basic solution saponification ester group, utilize aforementioned amido linkage to generate reagent (scheme 1), make gained 5-indole-carboxylic acid derivative 3 (v) with H
2N-R
6Coupling obtains the compound of formula I.
Embodiment
Now illustrate in greater detail the present invention by following indefiniteness embodiment.Institute responds and all carries out in nitrogen atmosphere or argon atmospher.Temperature is degree centigrade to provide.Flash chromatography carries out on silica gel.Unless otherwise indicated, otherwise the per-cent of solution or ratio by volume/volumeter.Electrospray mass spectrograph record is adopted in mass spectroscopy.Abbreviation used herein or symbol comprise:
DIEA: diisopropyl ethyl amine;
DMAP:4-(dimethylamino) pyridine;
DMSO: methyl-sulphoxide;
DMF:N, dinethylformamide;
Et: ethyl;
EtOAc: ethyl acetate;
Et
2O: ether;
HPLC: high pressure liquid chromatography;
iPr: sec.-propyl
Me: methyl;
MeOH: methyl alcohol;
MeCN: acetonitrile;
Ph: phenyl;
TBE: three-borate-EDTA;
TBTU:2-(1H-benzotriazole-1-yl)-N, N, N ', N '-tetramethyl-urea a tetrafluoro borate;
TFA: trifluoroacetic acid;
TFAA: trifluoroacetic anhydride;
THF: tetrahydrofuran (THF);
MS (ES): electrospray mass spectrum;
PFU: become the plate unit;
DEPC: diethylpyrocarbonate;
DTT: dithiothreitol (DTT);
EDTA: edetate;
HATU:O-(7-azepine benzo triazol-1-yl)-N, N, N
/, N
/-tetramethyl-urea hexafluorophosphate;
BOP: benzotriazole-1-base oxygen base-three (dimethylamino) Phosphonium hexafluorophosphate;
EDAC: see ECD;
DCC:1,3-dicyclohexyl phosphinylidyne diimine
The HOBt:1-hydroxybenzotriazole;
ES
+: electrospray (positive ionization);
ES
-: electrospray (negative electricity from);
DCM: methylene dichloride;
TBME: t-butyl methyl ether;
TLC: thin-layer chromatography;
AcOH: acetate;
EtOH: ethanol;
DBU:1,8-diazabicylo [5.4.0] 11 carbon-7-alkene;
BOC: tert-butoxycarbonyl;
Cbz: carbobenzoxy-(Cbz);
iPrOH: Virahol;
The NMP:N-methyl-2-pyrrolidone;
EDC:1-(3-dimethylaminopropyl)-3-ethyl phosphinylidyne diimmonium salt hydrochlorate;
RNAsin: ribonuclease inhibitor, sell by Promega company;
Tris:2-amino-2-hydroxymethyl-1, ammediol;
UMP: uridine 5 '-monophosphate;
UTP: uridine 5 '-triphosphate; And
IPA: isopropyl acetate.
The method of the synthetic representative compounds of the present invention of embodiment 1-22 explanation.
Embodiment 1
3-amino-4-iodobenzoic acid methyl esters:
(13.35g 50.8mmol) is added among the MeOH (150mL) and adds SOCl with 3-amino-4-iodobenzoic acid
2(4.8mL, 65.8mmol, 1.3 equivalents).This mixture was refluxed 3 hours, under reduced pressure remove volatile matter then.Residuum and MeOH coevaporation three times are then at evaporated in vacuo dry (15.23g).3-trifluoroacetamido-4-iodobenzoic acid methyl esters:
With derive from above-mentioned anils (14.53g, 52mmol) be dissolved in DCM (200mL) and add TFAA (15mL, 104mmol).The solution backflow of this mulberry is spent the night.Under reduced pressure removing volatile matter, and make residuum pass through short silicagel pad, is eluent with DCM.Obtain required product, it is peach solid (13.81g).
4-phenylacetylene base-3-(2,2,2-three fluoro-acetylamino)-methyl benzoate:
With derive from above-mentioned iodide (0.742g, 2mmol), phenylacetylene (0.37mL, 3.9mmol, 1.7 equivalents) and Et
3N (6mL) is added in the exsiccant flask under argon atmospher.Add PdCl
2(PPh
3)
2(0.241g 0.3mmol) also at room temperature stirs this mixture, till judging complete reaction by HPLC (～5 hours).Under reduced pressure, the reaction mixture volume is concentrated half, and water (80mL) dilution.(3 * 100mL) extractions, organic extract liquid is used 5%HCl (100mL) again with 5%HCl (100mL), and uses salt solution (40mL) washing with EtOAc with this mixture.Through MgSO
4After the drying, residuum carries out purifying by flash chromatography, is eluent with the 20%EtOAc-hexane, obtains the alkynes of required cross-coupling, and it is brown solid (0.442g).
3-(cyclohexenyl)-2-phenylindone 6-carboxylate methyl ester:
The anhydrous K that in flame-dried flask, adds finely powdered
2CO
3(0.153g, 1.1mmol) and derive from above-mentioned alkyne derivatives (0.390g, 1.1mmol).Adding exsiccant DMF (4mL) also outgases this suspension with argon gas stream.Add the enol triflate (0.802g, 3.3mmol, 3 equivalents) that comes from pimelinketone, it is according to A.G.Martinez, people such as M.Hanack (J.Heterocyclic Chem.1988,25,1237) suitable with it method prepares in described method or the document, adds Pd (PPh then
3)
4(0.086g 0.07mmol), and at room temperature stirred this mixture 8 hours.Under reduced pressure remove DMF, residuum carries out purifying by flash chromatography, is eluent (0.260g) with DCM.
3-cyclohexyl-2-phenylindone-6-carboxylate methyl ester:
In a usual manner, make solvent, by 20%Pd (OH) with MeOH
2Above-mentioned material hydrogenation (1atm H will be derived from
2Gas).After the filtration catalizer, separate required hexanaphthene indoles.
3-cyclohexyl-2-phenylindone-6-carboxylic acid:
(0.154g 0.15mmol) refluxes in the mixture of MeOH (10mL) and 2N NaOH (6mL) and spends the night, till HPLC analysis revealed complete hydrolysis with deriving from above-mentioned methyl ester.Be cooled to after the room temperature, add 2N HCl (5mL), adding AcOH to pH then is 7.Under reduced pressure remove MeOH, add water (50mL), product extracts with EtOAc.Extraction liquid water and salt water washing, and dry (MgSO
4).Under reduced pressure remove volatile matter, obtain the indole-carboxylic acid of title, it is the solid (0.149g) of light orange.
According to identical method, but replace phenylacetylene, obtain 3-hexanaphthene-2-(2-pyridyl) Indole-6-carboxylic acid with the 2-ethynyl pyridine.
Embodiment 2:
3-cyclohexenyl-6-indole-carboxylic acid:
In the round-bottomed flask of 12L, load onto reflux exchanger and mechanical stirrer, and with system's nitrogen purging.6-indole-carboxylic acid (300.00g, 1.86 moles, 3 equivalents) is added in the flask, adds MeOH (5.5L) then.After stirring 10 minutes under the room temperature, and the adding pimelinketone (579mL, 5.58mole).In 10 minutes, piecemeal adds methanol solution of sodium methylate (25%w/w, 2.6L, 11.37 moles, 6.1 equivalents).This mixture was refluxed 48 hours.Be cooled to after the room temperature, add water (4L) and under reduced pressure remove methyl alcohol.Remaining water with dense HCl (～1.2L) to be acidified to pH be 1.The little yellow mercury oxide of filter collection gained washes with water, and 50 ℃ of following vacuum-dryings.Obtain required cyclohexane derivant, it is beige solid (451.0g, 100% productive rate).
3-cyclohexyl-6-indole-carboxylic acid:
With deriving from above-mentioned unsaturated derivative under the 55psi hydrogen pressure, in 20%Pd (OH)
2/ C (10.25g) goes up hydrogenation 20 hours, and the THF-MeOH (2.5L) that uses 1: 1 is as solvent.After the filtration catalizer, under reduced pressure remove volatile matter, residuum is pulverized with hexane.Filter collection beige solid is used hexane wash, and vacuum-drying (356.4g, 78% productive rate).
3-cyclohexyl-6-indole-carboxylic acid methyl esters:
In the three-necked flask of 5L, load onto reflux exchanger and mechanical stirrer, and with system's nitrogen purging.(300.00g 1.233mole) is added in the flask, and makes it to be suspended among the MeOH (2L) with deriving from above-mentioned indole-carboxylic acid.Thionyl chloride (5mL, 0.0685 mole, 0.05 equivalent), and with this mixture backflow 48 hours.Under reduced pressure remove volatile matter, residuum is pulverized with hexane, obtains beige solid, with this solid with hexane wash and vacuum-drying (279.6g, 88% productive rate).
2-bromo-3-cyclohexyl-6-indole-carboxylic acid methyl esters:
Revise the method (Tet.Lett.1997,38,3871) of L.Chu, (4.65g 18.07mmol) is dissolved in THF (80mL) and CHCl with 3-cyclohexyl-6-indole-carboxylic acid methyl esters
3In the mixture (80mL).This solution is cooled off in ice bath, and add pyridinium bromide perbromide (pyridine tribromide, 7.22g, 22.6mmol, 1.25 equivalents).After 0 ℃ is stirred 1.5 hours down, judge according to TLC whether reaction is complete.Use CHCl
3(200mL) 1M NaHSO is used in dilution
3(2 * 50mL), saturated NaHCO
3The aqueous solution (2 * 50mL) and salt solution (50mL) washing.Through Na
2SO
4After the drying, removal of solvent under reduced pressure, residuum TBME-hexane crystallization.The 2-bromo indole derivative that the filter collection is required is with hexane wash and dry (3.45g).Mother liquid evaporation obtains red solid, and it carries out purifying by flash chromatography, utilizes the hexane solution of 15%EtOAc, obtains the pure substance of 3.62g in addition.Ultimate production is 5.17g (85% productive rate).
Embodiment 3:
The general method of the Suzuki cross-coupling of aryl and heteroaryl boric acid and 2-bromo indole derivative:
The cross-coupling of aryl/hetaryl boric acid or (boronic acid) or ester derivative and 2-bromo indole 2-bromo indole as described in example 2 above can utilize any variant of the known standard metal catalysis of the those skilled in the art described in document Suzuki cross-coupling reaction to carry out.The following example is used to illustrate this method, and right and wrong are determinate.
3-cyclohexyl-2-furans-3-base-1H-indole-6-carboxylic methyl ester:
With the 2-bromo indole of embodiment 2 (8.92g, 26.5mmol), 3-furans boric acid (B.P.Roques et al., J.Heterocycl.Chem.1975,12,195; 4.45g, 39.79mmol, 1.5 equivalents) and LiCl (2.25g, 53mmol, 2 equivalents) be dissolved in the mixture of EtOH (100mL) and toluene (100mL).The Na that adds 1M
2CO
3(66mL 66mmol) and with this mixture uses argon-degassed 45 minutes to the aqueous solution.Add Pd (PPh
3)
4(3.06g, 2.65mmol, 0.1 equivalent) also stirs this mixture and to spend the night under 75～85 ℃ and argon atmospher.Under reduced pressure remove volatile matter, residuum is dissolved in EtOAc (500mL) again.With this solution with water, saturated NaHCO
3(100mL) and salt solution (100mL) washing.Stand-by MgSO
4After the mixture drying of decolorizing charcoal, filter this mixture and concentrating under reduced pressure.Remaining oily matter is pulverized with the mixture of TBME (20mL) and hexane (40mL), cool off in, and filter collection precipitated solid is washed with the hexane solution of cold 25%TBME, and drying (3.09g).Filtrate and the washing lotion that derives from above-mentioned grinding are merged, concentrate and carry out purifying, adopt the EtOAc hexane solution of 10-25%, obtain the 4.36g product extraly by flash chromatography.The ultimate production of the 2-of embodiment 3 (3-furyl) indoles is 8.25g.
Embodiment 4:
3-cyclohexyl-1-methyl-6-indole-carboxylic acid methyl esters:
3-cyclohexyl-6-indole-carboxylic acid methyl esters (150.00g, 0.583 mole) of embodiment 2 is added in the three-necked flask of 3L, and this flask is equipped with mechanical stirrer and uses nitrogen purging.Add DMF (1L) and solution is cooled off in ice bath.One aliquot, one aliquot ground adds NaH (60% oil suspension, 30.35g, 0.759 mole, 1.3 equivalents) (15 minutes), and this mixture was stirred 1 hour under cooling.One aliquot, one aliquot ground adds methyl iodide (54.5mL, 0.876 mole, 1.5 equivalents), and keeping internal temperature is 5～10 ℃.Reaction mixture at room temperature stirred spend the night.Make the reaction cancellation by adding ice-water (3L), generate creamy throw out simultaneously.This material of filter collection washes with water, and in 45 ℃ of following vacuum-dryings (137.3g, 86% productive rate).
2-bromo-3-cyclohexyl-1-methyl-6-indole-carboxylic acid methyl esters:
Be added in the three-necked flask of 5L deriving from above-mentioned 1-methyl indole derivatives (136.40g, 0.503 mole), this flask is equipped with mechanical stirrer and uses nitrogen purging.Add CHCl
3(750mL) and THF (750mL), and with this solution be cooled to 0 ℃.One aliquot, one aliquot ground adds pyridine tribromide (pyridinium bromide perbromide, 185.13g, 0.579 mole, 1.15 equivalents), and this mixture was stirred 1 hour down at 0 ℃.Removal of solvent under reduced pressure at room temperature, and residuum is dissolved in EtOAc (3L).With this solution with water and salt water washing, dry (decolorizing charcoal/MgSO
4) and concentrating under reduced pressure.Be suspended in residuum among the TBME and be heated to 50 ℃.This suspension stored in refrigerator spend the night, and the creamy crystallized product of filter collection.It is washed and vacuum-drying (134.3g, 76% productive rate) with TBME.
Embodiment 5:
Cyclohexyl-methyl-tributyl tin alkyl-1H-indole-6-carboxylic methyl ester:
Under argon atmospher, (2.70g 7.71mmol) is dissolved in exsiccant THF (40mL) and this solution is cooled to-78 ℃ with the bromo indole derivative of embodiment 4.In 15 minutes, drip the hexane solution (1.4M, 6.90mL, 9.64mmol, 1.25 equivalents) of n-BuLi, and continue at low temperatures to stir 75 minutes.In 5 minutes, in gained suspension, add n-Bu
3SnCl (2.93mL, 10.8mmol, 1.4 equivalents).Suspension dissolves, and gained solution was stirred 1 hour down at-78 ℃.Reaction mixture is warming up to room temperature, and under reduced pressure removes THF.Residuum is dissolved in TBME (150mL),, and uses MgSO with salt solution-water washing of 1: 1
4Dry.This material carries out purifying by silica gel chromatography, described silica gel in advance by with 5% Et
3The N hexane solution mixes and activates.Identical solvent is also as the stratographic eluent.Separate the stannane of title, it is xanchromatic oily matter (3.42g, 79% productive rate).
Embodiment 6:
The general method of the 2-stannane indoles of embodiment 5 and the halid Stille cross-coupling of aryl/hetaryl: aryl/hetaryl halogenide or class halogenide (preferred bromide, iodide and triflate) with the cross-coupling of the stannane derivative of embodiment 5, can utilize any variant of the standard metal catalysis Stille cross-coupling reaction described in the document to carry out.The following example is used to illustrate this method.
3-cyclohexyl-1-methyl-2-pyridine-2-base-1H-indole-6-carboxylic methyl ester:
(3.42g 6.1mmol) is dissolved in DMF (10mL), and adds CuI (0.116g with the stannane derivative of embodiment 5,0.61mmol, 0.1 equivalent), LiCl (0.517g, 12.21mmol, 2 equivalents), triphenylphosphine (0.320g, 1.22mmol, 0.2 equivalent) and 2-bromopyridine (0.757mL, 7.94mmol, 1.3 equivalents).With this solution with the argon gas stream degassing (30 minutes) and add Pd (PPh
3)
4(0.352g, 0.31mmol, 0.05 equivalent).Use again after the argon purge 10 minutes, stir with the solution heating and under 100 ℃ and argon atmospher and spend the night.DMF is removed in decompression then, and residuum is dissolved in EtOAc (150mL).This solution is washed and uses MgSO with 1N NaOH (25mL) and salt solution (25mL)
4Dry.Removal of solvent under reduced pressure, residuum carries out purifying by flash chromatography, uses CHCl
3And the EtOAc of 5-10% subsequently is in CHCl
3In eluant solution (1.516g, 71% productive rate).
Embodiment 7:
The general method of the Stille cross-coupling of 2-bromo indole and aryl or heteroaryl stannane:
3-cyclohexyl-1-methyl-2-pyridine-2-base-1H-indole-6-carboxylic methyl ester:
With the 2-bromo indole derivative of embodiment 4 (0.150g, 0.428mmol) and 2-trimethylammonium stannyl thiophene (S.F.Thames et al., J.Organometal.Chem.1972,38,29; 0.150g, 0.61mmol, 1.4 equivalents) be dissolved in the dry THF (7mL) in the sealing test tube, and this solution was outgased 30 minutes with argon gas stream.Add Pd (Cl)
2(PPh
3)
2(0.018g, 0.026mmol, 0.06 equivalent) seals test tube then.This solution was heated 40 hours down at 80 ℃.Reaction mixture is cooled to room temperature, adds EtOAc (10mL) and filters this suspension.After the evaporating solvent, make residuum experience described reaction conditions 20 hours more again, adopt new 2-stannyl thiophene (0.150g, 0.61mmol) and catalyzer (0.020g).Be cooled to room temperature and also cross after the filter solid, evaporating solvent, residuum carries out the CHCl that purifying adopts 15-100% by flash chromatography
3Hexane solution is eluent (0.133g, 88% productive rate).
Can utilize identical method that the stannane derivative is coupled on the 2-bromo indole of embodiment 2.
Embodiment 8:
2-aryl and 2-heteroaryl-alkylating general method of 6-indoxyl carboxylic acid ester N-:
3-cyclohexyl-1-methyl-2-pyridine-2-base-1H-indole-6-carboxylic methyl ester:
NaH (60% oil suspension, 0.186g, 4.64mmol, 1.5 equivalents) is deoiled to remove with hexane (20mL) washing, be suspended in again then among the DMF (5mL).In ice bath, be cooled to after 0 ℃, drip indole derivatives (1.000g, 3.09mmol) solution in DMF (3mL+2mL flushing) of embodiment 3.After stirring 15 minutes, piecemeal adds methyl iodide (0.385mL, 6.18mmol, 2 equivalents), and this mixture was stirred 2 hours under cooling, and at room temperature stirs 2 hours.Make the reaction cancellation by adding 1N HCl (1mL) then, and dilute with TBME (100mL).This solution is washed with 1N HCl (25mL), and dry (MgSO
4).Under reduced pressure remove after the volatile matter, residuum carries out purifying by flash chromatography, and adopting the EtOAc hexane solution of 5-10% is eluent, obtains inscribing compound, and it is white solid (0.903g, 86% productive rate).
Other N-alkyl indole derivatives among the present invention can utilize similar method, and (iBuI BnBr) prepares for EtI for example, iPrI by suitable electrophilic reagent.
Embodiment 9:
The 6-indoxyl carboxylic acid ester is corresponding free carboxy acid's general method for saponification extremely:
This method is applicable to indoles and N-skatol carboxylic acid ester.
3-cyclohexyl-1-methyl-2-pyridine-2-base-1H-Indole-6-carboxylic acid:
(1.517g 4.35mmol) is dissolved in DMSO (8mL) and add 5N NaOH (4.4mL) with the 6-indoxyl carboxylic acid ester of embodiment 6.This mixture was stirred 30 minutes down at 50 ℃.Then this solution is cooled to room temperature and drips water (15mL).The insoluble black impurity of filtering also drips AcOH (2mL) to filtrate.The white precipitate that the filter collection is generated washes with water and dry (1.37g, 94% productive rate).
Embodiment 10:
1-cyclohexyl-2-phenyl-1H-indole-5-carboxylic acid:
4-amino-3-iodobenzoic acid ethyl ester:
With the 4-subcutin (15.00g, 91mmol) and iodine (11.80g is 46.5mmol) with water (80mL) and chlorobenzene (4.60g, 41mmol) mixing.Stir this mixture, in 30 minutes, temperature progressively is increased to 90 ℃ simultaneously.Under 90 ℃ in 10 hours, add hydrogen peroxide (30%, 50mL).Under this temperature, after the restir 6 hours, cool off this mixture, and decant goes out solution from residual solid.This solid is dissolved in DCM, and this solution is used Sulfothiorine and salt water washing successively.Dry (MgSO
4) afterwards, removal of solvent under reduced pressure, the gained brown solid grinds with hexane, to remove two iodinating by products.Obtain required compound, it is brown solid (22.85g, 86% productive rate).
4-acetylaminohydroxyphenylarsonic acid 3-iodobenzoic acid ethyl ester:
(1.00g 3.44mmol) is dissolved in pyridine (5mL) and also this solution is cooled off in ice with deriving from above-mentioned aniline.Drip AcCl (0.32mL, 4.47mmol, 1.3 equivalents), and this mixture was stirred 1 hour at 0 ℃, at room temperature stirred 2 hours.Reaction mixture is diluted with 1NHCl, and with TBME (100mL) extraction product.Organic phase is washed with 1N HCl (50mL), dry (MgSO
4) and concentrate, obtaining desired substance, it is brown solid (1.121g, 97% productive rate).
2-phenyl-indole-5-carboxylic acid, ethyl ester:
According to people's such as A.Bedeschi method (Tet.Lett.1997,38,2307), make from above-mentioned acetanilide derivative (0.900g, 2.7mmol) with phenylacetylene (0.385mL, 3.5mmol, 1.3 equivalents) at PdCl
2(PPh
3)
2(10 moles of %) and CuI (10 moles of %) exist down, react in the mixture of Yu diox (5mL) and tetramethyl guanidine (5mL).After the purification by flash chromatography through the 15%EtOAc hexane solution, separate required 2-phenylindone (0.589g, 82% productive rate), it is a yellow solid.
1-hexamethylene-1-thiazolinyl-2-phenyl-1H-indole-5-carboxylic acid ethyl ester:
(0.265g 1.0mmol) is dissolved in DMF (2mL) and add cesium hydroxide monohydrate (0.208g, 1.2mmol, 1.2 equivalents) with deriving from above-mentioned 2-phenylindone derivative.Cool off this solution in ice bath and drip the solution (5 minute) of 3-bromine tetrahydrobenzene (0.193g, 1.2mmol, 1.2 equivalents) in DMF (1mL).This mixture was stirred 30 minutes down at 0 ℃.With reactant water (25mL) dilution, use Et
2O (2x 50mL) extraction, extraction liquid MgSO
4Dry.Solvent evaporated under reduced pressure obtains white foam (0.095g), and it need not purifying and promptly can be used for next step.
1-cyclohexyl-2-phenyl-1H-indole-5-carboxylic acid:
To derive from above-mentioned thick indoles by general method (1atm H
2Gas) in EtOH, pass through 20%Pd (OH)
2-carbon hydrogenation at room temperature 20 hours.After the filtration catalizer, EtOH is removed in decompression.Residuum is dissolved in the mixture of MeOH (1mL) and DMSO (1mL), and adds 5NNaOH (0.5mL).The stirring under 50 ℃ of this mixture is spent the night.With reaction mixture cooling and add water (10mL).After the 1NHCl acidifying, product extracts in Et
2Among the O (70mL), and dry this solution (Na
28O
4).Evaporating solvent obtains green residuum, its be required 1-cyclohexyl-2-phenylindone-5-carboxylic acid in 1,2: 1 the mixture (85mg) of 3-dicyclohexyl-2-phenylindone-5-carboxylic acid.
Embodiment 11:
1-cyclohexyl-3-methyl-2-phenyl-1H-indole-5-carboxylic acid:
2-phenyl-3-methyl-indole-5-carboxylic acid's ethyl ester:
According to the method (Tet.Lett.1997,38,2439) of H.-C.Zhang, (derive from embodiment 10,0.500g 1.72mmol) is dissolved in DMF (5mL) and add LiCl (0.073g, 1.72mmol, 1 equivalent), PPh with 4-amino-3-iodobenzoic acid ethyl ester
3(0.090g, 0.34mmol, 0.2 equivalent), K
2CO
3(1.188g, 8.6mmol, 5 equivalents) and phenyl propine (0.645mL, 5.76mmol, 3 equivalents).With this solution with argon-degassed 1 hour and add acid chloride (0.039g, 0.17mmol, 0.1 equivalent).This mixture was stirred 20 hours under 80 ℃ and argon atmospher.Extract with reaction mixture water (25mL) dilution and with EtOAc (50mL).Extraction liquid washs with salt solution (3x 25mL) and dry (MgSO
4).Concentrating under reduced pressure and the flash chromatography by the 10-15%EtOAc-hexane solution carry out purifying, obtain required 2-phenyl-3-skatole (0.275g, the component of polarity minimum) and 3-phenyl-2-methyl isomer (0.109g, the component that polarity is bigger).
1-(3-cyclohexenyl)-3-methyl-2-phenylindone-5-carboxylic acid, ethyl ester:
(0.264g 0.95mmol) is dissolved in DMSO (2mL) and add cesium hydroxide monohydrate (0.191g, 1.14mmol with deriving from the less isomer of above-mentioned polarity, 1.2 equivalent), add 3-bromine tetrahydrobenzene (1.2 equivalents are in the DMSO of 0.7mL for 0.183g, 1.14mmol) then.This mixture was at room temperature stirred 30 minutes.Add additional C sOH monohydrate (0.400g, 2.4 equivalents) and 3-bromine tetrahydrobenzene (0.400g, 2.4 equivalents), and restir 30 minutes.Two kinds of reagent that add same amount once more, and under room temperature restir 30 minutes, afterwards, reactant is diluted with 1N HCl (6mL) and water (20mL).Product extracts with TBME (100mL), dry (MgSO
4), and after concentrating under reduced pressure, residuum carries out purifying by flash chromatography, adopting the 5-10%EtOAc hexane solution is eluent.Obtain required N-alkylated indoles (0.130g).
1-cyclohexyl-3-methyl-2-phenylindone-5-carboxylic acid, ethyl ester:
Above-mentioned unsaturated product (1atm H will be derived from
2Gas) at room temperature in the usual way by 20% Pd (OH)
2Solution hydrogenation in EtOH 3 hours.
1-cyclohexyl-3-methyl-2-phenyl-1H-indole-5-carboxylic acid:
To derive from the mixture that above-mentioned hydrogenated products is dissolved in DMSO (2mL) and MeOH (2mL).
Adding 5N NaOH (0.5mL) also spends the night the stirring under 60 ℃ of this mixture.After water (40mL) dilution, water-phase product was with 1: 1 Et
2O-hexanes mixtures (50mL) washing, being acidified to pH with 1N HCl then is 1.(2 * 50mL) extractions, extraction liquid passes through Na to the free acid that is discharged with ether
2SO
4Carry out drying.Removal of solvent under reduced pressure obtains required indoles, and it is light brown solid (0.074g).
Embodiment 12:
2-bromo-3-cyclopentyl-1-Methyl-1H-indole-6-carboxylate methyl ester:
In being housed, the 3L three-necked flask of mechanical stirrer adds indoles 6-carboxylic acid (220g, 1.365 moles) and KOH particle (764.45g, 13.65 moles, 10 equivalents).Add water (660mL) and MeOH (660mL) and with this mixture heating up to 75 ℃.In 18 hours, drip cyclopentanone (603.7mL, 6.825 moles, 5 equivalents) with pump.With reaction mixture reheat 3 hours (judging with HPLC whether reaction is complete thereafter) and in 1 hour internal cooling to 0 ℃.The sedimentary sylvite of filter collection, (2 * 500mL) washings are to remove the product of cyclopentanone from condensation with TBME.Brown solid is dissolved in water (2.5L) again, and also (2 * 1L) wash with TBME with this solution.Being acidified to pH with dense HCl (425mL) is after 3, the cream-coloured throw out of filter collection, water (2 * 1L) washings and in 70 ℃ of vacuum-dryings.Weight of crude product is 275.9g (88.9% mass recovery) and has 85% homogeneity (HPLC).
Be dissolved in MeOH (750mL) and add 20% Pd (OH) deriving from above-mentioned crude product (159.56g, 0.70 mole)
2-carbon (8.00g).With the hydrogenation 18 hours under 50psi hydrogen in the Parr device of this mixture.After reaction is finished, by the diatomite filtration catalizer, and removal of solvent under reduced pressure.Gained brown solid under 70 ℃ and vacuum dry 12 hours.Obtain crude product (153.2g), it is a brown solid, and has 77% HPLC uniformity.
Thick 3-cyclopentyl Indole-6-carboxylic acid (74.00g, 0.323 mole) is added to 3L is equipped with in the three-necked flask of mechanical stirrer and thermometer.With system's nitrogen purging, and add dry DMF (740mL).After the material dissolution, add Anhydrous potassium carbonate (66.91g, 0.484 mole, 1.5 equivalents), and this mixture was stirred 5 minutes.Add methyl iodide (50mL, 0.807 mole, 2.5 equivalents) and also this mixture was stirred 5 hours, thereafter, the HPLC analysis revealed of reaction mixture, 97% changes into methyl esters.Reaction mixture is cooled off in ice bath, and an aliquot one aliquot ground adds sodium hydride (95%, do not have oil, 10.10g, 0.420 mole, 1.3 equivalents) (heat release: internal temperature rises 8～30 ℃) in 3 minutes.After stirring separately 15 minutes, remove cooling bath, and at room temperature continue to stir 1.5 hours, afterwards, do not observe any reaction progress (HPLC).Add other NaH (1.55g, 65mmol, 0.2 equivalent) and methyl iodide (1.0mL, 16mmol, 0.05 equivalent) again, and after stirring 15 minutes, judge reaction whether fully (96% N-methylates) by HPLC.Under vigorous stirring, with reaction mixture lentamente (2 minutes) pour in the water (4L), and after 10 minutes, be acidified to pH＜2 with dense HCl (85mL).This mixture was stirred 5 minutes, make remaining salt of wormwood and saleratus change into more diffluent muriate fully.With 4N NaOH (40mL) with pH regulator to～7, and this mixture at room temperature stirred spends the night.Filter collection throw out, water (600mL) washing is also dry under 60 ℃ and vacuum.Obtain crude product (79% HPLC uniformity), it is brown solid (72.9g).Grind composition with deriving from the MeOH of above-mentioned crude product, to remove a series of accessory impurity with minimum.Filter collects solid then, and it is dissolved among the hot EtOAc of minimum.Be cooled to after the room temperature, add hexane (5X volume), cool off in ice this mixture and filtration.Evaporate to dryness filtrate obtains required product then.(10.60g 41.2mmol) is dissolved in isopropyl acetate (150mL) and add sodium acetate (5.07g, 62mmol, 1.5 equivalents) with deriving from above-mentioned N-skatole.This suspension is cooled off in ice bath, and in 2 minutes, add bromine (2.217mL, 43.3mmol, 1.05 equivalents).The suspension of light amber becomes garnet (temperature rises to 13 ℃ by 5 ℃).It was stirred 1 hour at 0 ℃.Shown in the HPLC analysis,, make to react completely by adding other bromine (0.21mL, 4.2mmol, 0.10 equivalent).Make the reaction cancellation by adding 10% sodium sulfite aqueous solution (15mL) then, afterwards, add water (50mL) and K
2CO
3(10.6g, 1.8 equivalents) with in and HBr.Tell organic layer, sodium sulfite aqueous solution with 10% and K
2CO
3Solution washing and dry (MgSO
4).Removal of solvent under reduced pressure, residuum and TBME (75mL) condistillation obtains beige solid, and it is dried overnight (13.80g) under vacuum.Crude product was ground 30 minutes Yu Bingzhong cooling, and filter collection beige solid with ebullient MeOH (80mL).Product is dry (10.53g, 76% rate of recovery) under 60 ℃ and vacuum.
Embodiment 13
3-cyclopentyl-1-methyl-2-vinyl-1H-Indole-6-carboxylic acid:
To the 2-of embodiment 12 bromo indole derivative (2.044g, add in dry diox (20mL) solution 6.08mmol) the vinyl tributyl tin (1.954mL, 6.69mmol).With this solution by the foaming nitrogen degassing 15 minutes, add then two (triphenylphosphine) palladium (II) muriate (213.4mg, 0.304mmol) and with reaction mixture 100 ℃ of following heated overnight.Reaction mixture is also washed with hydration salt solution successively with the ether dilution.Conventional processing (MgSO
4, filter and concentrate) afterwards, residuum carries out flash chromatography (5cm, 10%AcOEt-hexane), obtains required compound (1.32g, 4.70mmol, 77% productive rate), and it is a white solid.
(153mg 0.54mmol) adds lithium hydroxide aqueous solution (226.6mg, 5.40mmol is in the water of 1.6mL) in the solution in the mixture of THF (2.8mL) and methyl alcohol (1.4mL) to deriving from above-mentioned ester.Reaction mixture was stirred 1.5 hours down and dilute with water at 50 ℃.Water layer is with the dilution of the HCl aqueous solution of 1M and use CH
2Cl
2Extract three times.Organic layer after the merging is water (2X) and salt water washing successively.(MgSO after the conventional processing
4, filter and concentrate), isolate required thick acid (150mg).
Embodiment 14
3-cyclohexyl-1-methyl-2-oxazole 5-base-1H-Indole-6-carboxylic acid:
To the bromide of embodiment 4 (1.00g, 2.855mmol) add in the solution in dry diox (10mL) the vinyl tributyl tin (917.8 μ L, 3.141mmol).By the foaming nitrogen degassing 15 minutes, (101mg 0.144mmol) and with this solution refluxed 7 hours to add two (triphenylphosphine) palladium (II) muriate then with this solution.Reaction mixture is diluted and water and salt water washing successively with ether.(MgSO after the conventional processing
4, filter and concentrate), residuum carries out flash chromatography (5cm, the AcOEt-hexane solution of the AcOEt to 10% of the AcOEt to 5% of hexane to 2.5%), obtains required compound (773mg, 2.60mmol, 91% productive rate), and it is a faint yellow solid.
(100mg 0.336mmol) in acetone (690 μ L), adds N-methylmorpholine N-oxide compound (NMMO in the mixture of the trimethyl carbinol (690 μ L) and water (690 μ L) successively to deriving from above-mentioned olefinic derivatives; 48mg, 0.410mmol) and the t-butanol solution (33 μ L) of 2.5% perosmic anhydride.Reaction mixture at room temperature stirred concentrated then in 3 days.Residue is dissolved in EtOAc than domestic water successively (2X) and salt water washing.(MgSO after the conventional processing
4, filter and concentrate), isolate thick glycol (117mg).Under 0 ℃, to derive from above-mentioned thick glycol (approximately 0.336mmol) in the mixture of THF (3.2mL) and water (3.2mL), add sodium periodate (86.2mg, 0.403mmol).Remove ice bath then, and reaction mixture was at room temperature stirred 1 hour 45 minutes.Add AcOEt then.Gained solution is used 10% aqueous citric acid solution successively, water, saturated NaHCO
3The aqueous solution, water (2X) and salt water washing.(MgSO after the conventional processing
4, filter and concentrate), isolate required thick aldehyde (92mg, 0.307mmol, 91% productive rate).
With derive from above-mentioned aldehyde mixture (25.8mg, 0.086mmol), Anhydrous potassium carbonate (12.4mg, 0.090mmol) and Tosmic (17.57mg 0.090mmol) refluxed in absolute MeOH (500 μ L) 2 hours.Add AcOEt then, and with this mixture water successively (2X) and salt water washing.(MgSO after the conventional processing
4, filter and concentrate), isolate required De Cu oxazole (28mg, 0.083mmol, 96% productive rate).
Derive from mutually above-mentioned ester (28mg, 0.083mmol) in THF (425 μ L), add in the mixture of MeOH (210 μ L) and water (250 μ L) lithium hydroxide (34.8mg, 0.830mmol).Reaction mixture at room temperature stirred spend the night, then dilute with water and with 1N HCl acidified aqueous solution.Water layer extracts with methylene dichloride (3X), and water (2X) and salt water washing successively.(MgSO after the conventional processing
4, filter and concentrate), the thick acid (30mg) of isolating title.
Embodiment 15
2-(the 1H-benzimidazolyl-2 radicals-yl)-3-cyclohexyl-1-Methyl-1H-indole-6-carboxylic acid:
To the aldehyde that derives from embodiment 14 (28mg, 0.094mmol) and 1, the 2-diaminobenzene (10.9mg, 0.101mmol) add in the mixture in acetonitrile (500 μ L) and DMF (200 μ L) chloranil (24.8mg, 0.101mmol).Reaction mixture was at room temperature stirred 3 days.Add AcOEt, and reaction mixture is used the NaOH aqueous solution (2X) of 1N, water (4X) and salt water washing successively.(MgSO after the conventional processing
4, filter and concentrate), residuum is implemented flash chromatography (1cm, 30%AcOEt-hexane), obtains required benzoglyoxaline ester derivative (11mg, 0.028mmol, 30% productive rate).
To above-mentioned ester (11mg, 0.028mmol) in THF (240 μ L), add in the mixture in MeOH (120 μ L) and the water (140 μ L) lithium hydroxide (11.7mg, 0.280mmol).Reaction mixture at room temperature stirred spend the night, dilute with water is with the HCl acidified aqueous solution of 1N then.Water layer extracts and water (2X) and salt water washing successively with methylene dichloride (3X).(MgSO after the conventional processing
4, filter and concentrate), separate the thick acid (9mg, 0.0241mmol, 86% productive rate) that obtains title.
Embodiment 16
3-cyclopentyl-1-Methyl-1H-indole-2,6-dicarboxylic acid 6-methyl esters:
Under 0 ℃, to the 3-cyclopentyl aldehyde (20mg for preparing in the mode that is similar to embodiment 15,0.07mmol) and 2-methyl-2-butene (541 μ L, 5.11mmol) the trimethyl carbinol (500 μ L) solution in add freshly prepd Textone (64.2mg be 0.711mmol) in the phosphate buffered saline buffer (NaH of 98mg
2PO
4In the water of 150 μ L) in solution.Reaction mixture was at room temperature stirred 45 minutes, add salt solution then.Water layer EtOAc extracting twice.Organic layer after the merging is used the HCl aqueous solution and the salt water washing of 0.5N successively.(MgSO after the conventional processing
4, filter and concentrate), obtain the required thick acid of 23.1mg, it is a yellow solid.
Embodiment 18
3-cyclopentyl-2-pyridine-2-base-cumarone-6-carboxylic acid:
2-bromobenzene benzofuran derivs (0.850g with embodiment 17,2.93mmol), 2-three (normal-butyl) stannyl pyridine (1.362g, 3.7mmol), triphenylphosphine (0.760g, 2.90mmol), lithium chloride (0.250g, 5.9mmol) and CuI (0.057g 0.3mmol) is dissolved in DMF (30mL), and with this mixture with bubbling argon-degassed 30 minutes.(0.208g 0.18mmol), and stirs this mixture under 100 ℃ and argon atmospher to add tetrakis triphenylphosphine palladium.After 19 hours, reactant is cooled to room temperature, pours in the water (70mL) and and extract with TBME.Organic phase water (2X) and salt water washing, dry (MgSO
4) and concentrate, the gained residuum carries out purifying by flash chromatography.Obtain required 2-(2-pyridyl) benzofuran derivative (0.536g, 63% productive rate), it is a white solid.(0.200g 0.694mmol) is suspended in dense H with deriving from above-mentioned nitrile
2SO
4(5mL), in the mixture of AcOH (4mL) and water (2mL).After refluxing 1.5 hours, TLC shows complete hydrolysis.This mixture is cooled off in ice, and the NaOH to pH that drips 10N then is 9.The water layer washed with dichloromethane, being acidified to pH with the HCl of 5N then is 6.Product extracts with EtOAc, dry (MgSO
4) and removal of solvent under reduced pressure.Obtain required carboxylic acid, it is a white solid.
Embodiment 19
2-bromo-3-cyclopentyl-benzo [b] thiophene-6-carboxylic acid, ethyl ester
At room temperature, (5.194g adds 1 in DMF 19.30mmol) (58.0mL) solution, 4-diazabicylo [2.2.2] octane (4.33g to the 3-of embodiment 17 bromo-6-pentamethylene carbonyl phenol, 38.60mmol) and dimethyl sulphide for urea chloride (4.77g, 38.6mmol).This mixture was at room temperature stirred 3 hours.It is 3 that this mixture is acidified to pH with 1N HCl, extracts with EtOAc then.Merge organic layer and use the salt water washing, and pass through MgSO
4Carry out drying.Thick mixture carries out purifying by the EtOAc/ hexane of silica gel plug and 3%, obtains the required thiocarbamate of 6.976g (100%), and it is colourless oily matter.
To derive from above-mentioned purified O-3-bromo-6-pentamethylene carbonyl N, (43.147g 121.1mmol) is heated to 180～190 ℃ internal temperature 5 hours to N-dimethyl thiocarbamate.Utilize TLC (20% EtOAc/ hexane: Rf0.6 (raw material), 0.5 (product)) to monitor reaction process.Crude product need not to be further purified and promptly can be used for next step reaction.
To derive from above-mentioned S-3-bromo-6-pentamethylene carbonyl N, N-dimethyl thiocarbamate is dissolved in MeOH (600mL), and (40.0g 714mmol) and with this mixture heating up refluxed 1.5 hours to add KOH.This mixture is cooled to room temperature, removes by rotary evaporation then and desolvate.Residuum is dissolved in water and is acidified to pH with the HCl of 6N is 3.It is extracted with EtOAc, and crude product carries out purifying by silica gel chromatography with 1～5% EtOAc/ hexane.Obtain the required thiophene derivant of 31.3g (91%), it is an xanchromatic oily matter.
(0.314g adds K in acetone 1.105mmol) (5.0mL) solution to deriving from above-mentioned 3-bromo-6-pentamethylene carbonyl thiophene
2CO
3(0.477g, 3.45mmol), add then ethyl bromoacetate (0.221g, 0.147mL, 1.33mmol).This mixture stirring is spent the night.With reaction mixture with filter paper filtering and concentrated filtrate.Carry out purifying by silica gel and with 5% EtOAc/ hexane, obtain 0.334g (82%) product, it is colourless oily matter.
Be dissolved in THF (12.0mL) with deriving from above-mentioned thick ester, at room temperature add the NaOH (5.0mL) of 1N.This mixture was at room temperature stirred 2～3 hours, perhaps until TLC show react completely till.Remove by rotary evaporation and to desolvate.Adding water and this mixture is acidified to pH with the HCl of 6N is 3, extracts with EtOAc then, uses the salt water washing, and uses MgSO
4Dry.Removal of solvent under reduced pressure, residuum need not purifying and can use.
In deriving from above-mentioned thick acid, add diacetyl oxide (16.0mL), add NaOAc (0.573g) then, and this mixture heating up backflow is spent the night.This mixture is cooled to room temperature and pours in the mixture of ice and toluene.Add 6N NaOH and reach about 7,, use the salt water washing, and use MgSO with the EtOAc extraction until pH
4Dry.Except that desolvating, residuum obtains the 6-bromo-3-cyclopentyl thionaphthene of 0.795g (80%) by silica gel hexane purifying by rotary evaporation, and it is colourless oily matter.
With derive from above-mentioned 6-bromo-3-cyclopentyl thionaphthene (0.723g, 2.57mmol) and cupric cyanide (0.272g, 3.04mmol) mixture heating up in DMF (1.4mL) refluxes and to spend the night.This mixture is cooled to room temperature and dilutes with EtOAc.The NH that adds 2N
4OH, and with this mixture stirring 10 minutes, filter by diatomite then.Water layer extracts with EtOAc.Merge organic layer, and use the salt water washing, use MgSO
4Drying, and removal of solvent under reduced pressure.Product need not purifying and can use.
With 3-cyclopentyl-6-cyano group thionaphthene (17.65g 77.65mmol) is dissolved in acetate (310mL), and at room temperature add bromine (49.64g, 310.6mmol).This mixture at room temperature stirred spend the night, and with HPLC monitoring reaction process.After reacting completely, in reaction mixture, add toluene (3 * 100mL), to remove acetate.With the crude product drying under reduced pressure, and need not to be further purified and to use.Be added to ethanol (150mL, sex change) and dense H with deriving from above-mentioned thick cyano derivative
2SO
4(45mL), and with this mixture heating up backflow 1～2 day.After reaction is finished (HPLC), reaction mixture is cooled to room temperature and pours in ice-water usefulness methylene dichloride (5 * 100mL) extractions, merging organic layer, NaHCO into 5%
3With the salt water washing.Removal of solvent under reduced pressure, residuum carries out purifying by the EtOAc/ hexane of silica gel and 1%.Concentrate collected cut, the residuum pulp is in methyl alcohol.Cross filter solid, and with ice-cold methanol wash, obtain the 15.9g pure ethyl ester in (58%, two step), it is flaxen solid.
Embodiment 20
3-cyclopentyl-2-pyridine-2-base-benzo [b] thiophene-6-carboxylic acid:
2-bromobenzene thiophthene (0.354g with embodiment 19,1.00mmol), 2-three (normal-butyl) stannyl pyridine (0.442g, 1.2mmol), triphenylphosphine (0.262g, 1.00mmol), lithium chloride (0.085g, 2.0mmol) and CuI (0.019g 0.1mmol) is dissolved in DMF (10mL), with this mixture by the argon gas bubbling degassing 30 minutes.(0.069g 0.06mmol), and will stir under 100 ℃ of this mixture soaps and the argon atmospher to add tetrakis triphenylphosphine palladium.After 24 hours, reactant is cooled to room temperature, pours in the water (70mL) and and extract with TBME.Organic phase water (2X) and salt water washing, dry (MgSO
4) concentrate, the gained residuum carries out purifying by flash chromatography.Obtain required 2-(2-pyridyl) benzothiophene ester (0.197g, 56% productive rate), it is flaxen waxy solid.
Use the NaOH hydrolysis in the usual way with deriving from above-mentioned ester, obtain the acid of title, it can directly use, and also can carry out purifying by HPLC and flash chromatography.
This acid can be according to general method and sulfonamide derivatives coupling described in the following embodiment 37.
Embodiment 21
3-cyclopentyl-2-furans-3-base-benzo [b] thiophene-6-carboxylic acid:
With the 2-bromobenzene thiophthene ester of embodiment 19 and the 3-furans boric acid coupling described in the embodiment 3, obtain required 2-(3-furyl) benzothiophene ester, productive rate is 85%.At room temperature use this ethyl ester of NaOH saponification, obtain the carboxylic acid derivative of title.
Embodiment 22
3-cyclohexyl-1-methyl-2-phenyl-1H-pyrrolo-[2,3, b] pyridine-6-carboxylic acid:
7-azaindole (15.00g .127 mole) is dissolved in MeOH (330mL) and adds sodium methylate (25%w/w in MeOH, 172mL, 0.753 mole) and pimelinketone (52.86mL, 0.51 mole).This mixture was refluxed 60 hours, then concentrating under reduced pressure.After cooling off in ice-water, it is 8 that reaction mixture is acidified to pH with 3N HCl, and filter collection precipitated solid.Product washes with water, grinds with the TBME-hexane, and by coming dry (19.8g) with the methylbenzene azeotropic distillation.
(15.00g 75.65mmol) is dissolved in the mixture of EtOH (130mL) and THF (30mL), and adds 20% Pd (OH) with above-mentioned substance
2-carbon (1.30g).With the H of this mixture at 1atm
2Hydrogenation is 24 hours under the gas, thereafter, adds extra catalyzer (1.30g) immediately.At H
2After additionally stirring 16 hours under the atmosphere, filtration catalizer, and with this solution decompression evaporation, the gained residuum is pulverized with TBME, obtains amber solid (13.9g).
With above-mentioned 7-azaindole derivatives (7.50g, 37.45mmol) be dissolved in DME (130mL) and add metachloroperbenzoic acid (12.943g, 60.0mmol).After stirring 2 hours, under reduced pressure remove volatile matter, residuum is suspended in the water (100mL).By adding saturated Na under the vigorous stirring
2CO
3The aqueous solution with this mixture alkalize to pH be 10.Filter collects solid then, water and a small amount of TBME washing, and dry (7.90g).
With above-mentioned thick N-oxide compound (4.00g, 18.49mmol) be dissolved in DMF (350mL) and in 5 minutes an aliquot one aliquot ground add NaH (60% suspension, 1.52g, 38mmol).This mixture was stirred 30 minutes, and (1.183mL 19mmol) is added drop-wise in this suspension with methyl iodide in 20 minutes.After at room temperature stirring 3 hours, get nowhere again by HPLC assay determination reaction.Reaction mixture poured in the water and with EtOAc extraction three times.Extraction liquid salt water washing, dry (MgSO
4) and evaporation, obtaining amber solid (3.65g, 60% NMR uniformity), this solid can use immediately, need not purifying.
(0.80g 3.47mmol) is dissolved in MeCN (10mL) with above-mentioned crude product.Add triethylamine (1.13mL, 8.1mmol), add then trimethylsilyl cyanide (2.13mL, 16mmol).Then this solution was refluxed 19 hours.Be cooled to after the room temperature, by slow adding NaHCO
3The aqueous solution makes the reaction cancellation, and product extracts with EtOAc.Extraction liquid salt water washing, dry (MgSO
4) and concentrate, residuum carries out purifying (0.285g) by flash chromatography on silica gel with 15% EtOAc-hexane.(0.300g 1.254mmol) is suspended among the EtOH (15mL) and fed the hydrogen chloride gas bubbling 15 minutes, obtains clear soln with this nitrile.Then this solution was refluxed 1.5 hours, till TLC shows that raw material transforms fully.Be cooled to after the room temperature, under reduced pressure remove volatile matter, residuum is dissolved in EtOAc.The salt water washing of this solution, dry (MgSO
4) and concentrate.Residuum carries out purifying (the EtOAc-hexane of 15-20%) by flash chromatography on silica gel, obtains required ethyl ester, and it is flaxen jelly (0.227g).
With above-mentioned ester (0.100g, 0.35mmol) be dissolved in THF (4mL) and add pyridine perbromide hydrobromate (0.200g, 0.532mmol).This mixture is stirring 16 hours (transformation efficiency＞80%) in the bottle of sealing under 65 ℃.Evaporate this solution, residuum is absorbed among the EtOAc.With this solution with water and salt water washing, dry (MgSO
4) and concentrate.Crude product carries out purifying through flash chromatography on silica gel (15% EtOAc-hexane).
With above-mentioned bromide (0.100g, 0.274mmol), phenyl-boron dihydroxide (0.049g, 0.4mmol) and lithium chloride (0.019g 0.45mmol) is dissolved in toluene (2mL), EtOH (2mL) and 1M Na
2CO
3In the mixture (0.43mL).Make argon gas pass through this solution so that this mixture degassing 30 minutes, and add tetrakis triphenylphosphine palladium (0.035g, 0.03mmol).This mixture was refluxed 18 hours, thereafter, add once more catalyzer (0.035g, 0.03mmol).After refluxing 2 hours in addition, EtOH is removed in decompression again.Residuum is dissolved in EtOAc, with this solution with 10% the HCl aqueous solution and salt water washing, and dry (MgSO
4).Under reduced pressure remove volatile matter, the orange jelly of gained carries out purifying (0.105g, crude product) by flash chromatography on silica gel with 20% EtOAc-hexane.
(0.100g 0.276mmol) is dissolved in the mixture of THF (2mL) and EtOH (2mL) with deriving from above-mentioned partially purified ester.Adding 1N NaOH (2.8mL) also at room temperature stirred this mixture 4 hours.Under reduced pressure remove volatile matter, residuum dilutes with 10% the HCl aqueous solution.Product extracts with EtOAc (3X), dry (MgSO
4), evaporate and prepare HPLC and carry out purifying by anti-phase, obtain title compound.
Embodiment 23: the activity that suppresses NS5B RNA RNA-dependent polysaccharase
The activity that suppresses HCV RNA dependency polysaccharase (NS5B) with following experimental test The compounds of this invention:
Substrate is:
12 Nucleotide RNA widow-uridylic acids (or widow-uridine-single phosphoric acid) are primer (oligo-U), at its free 5 ' C position biotin modification;
The complementary polyadenylic acid (or adenosine monophosphate) of different lengths (1000-10000 Nucleotide) is template (polyA); And
UTP-[5，6
3H]。
With UMP-[5,6
3H] insert in the chain that goes out by the oligo-U primer extension, measure polymerase activity with this.
3The reaction product of H-mark is caught by the SPA pearl of streptavidin by bag, and quantitative on TopCount.
All solution [are added to 2ml DEPC in the 1L MilliQ water by the MilliQ water that DEPC handles; This mixture is acutely shaken with dissolving DEPC, then 121 ℃ of autoclavings 30 minutes] preparation.Enzyme: total length HCV NS5B (SEQ ID NO.1) is purified out with the form of the terminal six Histidine fusion roteins of N-from the insect cell of baculovirus infection.This enzyme can be preserved in-20 ℃ preservation damping fluid (as follows).Under these conditions, find that it can keep activity 6 months at least.
Substrate: biotinylation oligo-U
12Primer, Poly (A) template, and UTP-[5,6
3H] all be dissolved in water.These solution can be-80 ℃ of preservations.
Test damping fluid: 20mM Tris-HCl pH 7.5
5mM MgCl
2
25mM KCl
1mM EDTA
1mM DTT
NS5B preserves damping fluid: 0.1 μ M NS5B
25mM Tris-HCl pH 7.5
300mMNaCl
5mM DTT
1mM EDTA
0.1% dodecyl maltoside
30% glycerine
Testing compound mixture: be about to begin before the test testing compound of the present invention is dissolved in the test damping fluid that contains 15%DMSO.
Substrate mixture: before being about to begin test, various substrates are mixed in the test damping fluid according to following concentration:
| Component | Concentration in the substrate mixture | Final concentration in the test damping fluid |
| RNAsin TM | 0.5U/μL | 1.67U/μL |
| Vitamin H-widow-U 12Primer | 3ng/μL | 1ng/μL |
| The PolyA template | 30ng/μL | 10ng/μL |
| UTP-[5，6- 3H] 35Ci/mmol | 0.025μCi/μL | 0.0083μCi/μL 0.25μM |
| UTP | 2.25μM | 0.75μM |
Enzyme mixture: before being about to begin test, in the test damping fluid, prepare RNA polymerase (NS5B) mixture according to following explanation:
| Component | Concentration in the mixture |
| Tris-HCl，pH 7.5 | 20mM |
| MgCl 2 | 5mM |
| KCl | 25mM |
| EDTA | 1mM |
| DTT | 1mM |
| The dodecyl maltoside | 1％ |
| NS5B | 30nM |
Scheme:
Analytical reaction are at Microfluor
TMWhite " U " shape base plate (Dynatech
TMCarry out #7105), add successively:
20 μ L testing compound mixtures;
20 μ L substrate mixture; With
20 μ L enzyme mixtures
(final [the NS5B]=10nM in the test; Final [dodecyl maltoside]=0.33% in the test; Final DMSO=5% in the test).
Reactant was room temperature insulation 1.5 hours.Add STOP solution (20 μ L; 0.5M EDTA, 150ng/ μ l tRNA), add again 30 μ l streptavidin bag quilts the PVT pearl (8mg/ml is in 20mM Tris-HCl, and pH 7.5,25mM KCl, 0.025%NaN3).Then with flat board vibration 30 minutes.(70 μ L 5M), make CsCl concentration reach 1.95M to add CsCl solution.This mixture was left standstill 1 hour.These pearls are at Hewlett Packard TopCount
TMInstrument is counted in order to following scheme:
Data pattern: counting/minute
Scintiloscope: liq/plast
Energy region: low
Effectiveness pattern: normal
Interval: 0-50
Counting time-delay: 5 minutes
Gate time: 1 minute
The result who estimates: 6000cpm/ hole
The 200cpm/ hole does not contain the contrast of enzyme.
According to the The above results that test compounds obtains at 10 kinds of different concns, drawing standard concentration-% suppresses curve, by analyzing the IC that determines compound of the present invention
50For some compounds, at two some assessment IC
50
The specificity that embodiment 24:NS5B RNA RNA-dependent polysaccharase suppresses
Compound of the present invention is to the inhibition activity of polio virus rna RNA-dependent polysaccharase and calf thymus DNA RNA-dependent polymerase II, test according to the pattern identical with the HCV polysaccharase, difference is to replace HCV NS5B polysaccharase with another kind of polysaccharase.
Embodiment 25: the HCV rna replicon based on cell is analyzed
Cell cultures
The Huh7 of energy stable maintenance subgene group HCV replicon is according to the described foundation of document (Lohman et al., 1999.Science 285:110-113), with its called after S22.3 clone.The S22.3 cell is kept in the Dulbecco modified form Earle substratum (DMEM) of having added 10%FBS and 1mg/mL Xin Meisu (standard medium).During the analysis, adopt the DMEM substratum (test medium) contain 10%FBS and 0.5%DMSO but to lack Xin Meisu.Add before 16 hours of compound,, and be diluted to 50000 cell/ml with standard medium with S22.3 cell trypsin treatment.In each hole of 96 orifice plates, add 200 μ L (10000 cells).Then with flat board at 37 ℃, 5%C0
2Insulation is up to next day.
Reagent and material:
| Product | Company | Catalog number (Cat.No.) | Store |
| DMEM | Wisent Inc. | 10013CV | 4℃ |
| DMSO | Sigma | D-2650 | RT |
| Dulbecco′s PBS | Gibco-BRL | 14190-136 | RT |
| Foetal calf serum | Bio-Whittaker | 14-901F | -20℃/4℃ |
| Xin Meisu (G418) | Gibco-BRL | 10131-027 | -20℃/4℃ |
| Trypsinase-EDTA | Gibco-BRL | 25300-054 | -20℃/4℃ |
| The 96-orifice plate | Costar | 3997 | RT |
| PVDF 0.22 μ m filter membrane | Millipore | SLGV025L S | RT |
| Polypropylene deep hole titer plate | Beckman | 267007 | RT |
The preparation of test compound
10 μ L test compounds (in 100%DMSO) are added in the 2ml test medium, and making the DMSO final concentration is 0.5%, and this solution is used supersound process 15 minutes, filters 0.22 μ M Millipore filter membrane.900 μ l are transferred in the A round of polypropylene deep hole titer plate.Comprise the test medium (containing 0.5%DMSO) of 400 μ L sample aliquot in H row's the hole at B, with they preparation serial dilution things (1/2), method is to get 400 μ l to transfer to another row (H row does not contain compound) from a row at every turn.
Testing compound is applied to cell
From containing 96 orifice plate sucking-off cell culture mediums of S22.3 cell.The test medium that 175 μ L is contained corresponding extent of dilution testing compound from each hole of compound flat board is transferred in the respective aperture on the cell cultures flat board (H row is as " unrestraint contrast ").With the cell cultures flat board at 37 ℃, 5%C0
2Middle insulation 72 hours.
Extract cell total rna
Be incubated after 72 hours, with RNeasy 96 test kit (Qiagen
, RNeasy Handbook.1999.) and from the S22.3 cell of 96 orifice plates, extract cell total rna.In brief, test medium is thoroughly removed from cell, in each hole of 96 porocyte culture plates, added the RLT damping fluid (Qiagen that 100 μ L contain the 143mM beta-mercaptoethanol
).With the gentle vibration of plate 20 seconds.In each hole, add 100 μ L, 70% ethanol again, mix by piping and druming.Take out lysate, be loaded into as for Qiagen
RNeasy 96 (the Qiagen at Square-Well Block top
) the hole in.With RNeasy 96 dull and stereotyped sealings, the Square-Well Block that will have RNeasy 96 flat boards fixes with fixer, places the turning barrel of 4K15C whizzer with adhesive tape.With sample room temperature with 6000rpm (～5600xg) centrifugal 4 minutes.Remove adhesive tape from flat board, in each hole of RNeasy 96 flat boards, add 0.8ml BufferRW1 (Qiagen
RNeasy 96kit).Use new rubber belt sealing with RNeasy 96 is dull and stereotyped again, room temperature centrifugal 4 minutes with 6000rpm.RNeasy 96 flat boards are placed the top of another clean Square-WellBlock, remove adhesive tape, every hole adds 0.8ml Buffer RPE (Qiagen
RNeasy 96kit).Use new rubber belt sealing with RNeasy 96 is dull and stereotyped, room temperature centrifugal 4 minutes with 6000rpm.Remove adhesive tape, in each hole of RNeasy 96 flat boards, add another part 0.8ml Buffer RW1 (Qiagen
RNeasy 96kit).Use new rubber belt sealing with RNeasy 96 is dull and stereotyped again, room temperature centrifugal 10 minutes with 6000rpm.Remove adhesive tape, RNeasy 96 flat boards are placed on the support that contains 1.2-mL collection small test tube.Every hole adds the water that 50 μ L do not contain RNase, in case eluted rna, with the new rubber belt sealing of flat board, room temperature insulation 1 minute, room temperature 6000rpm is centrifugal 4 minutes then.Add 50 μ l once more and do not contain the water of RNase, repeat elution step.The small test tube that will contain cell total rna is kept at-70 ℃.
Quantitative cell total rna
RNA is at STORM
(the Molecular Dynamics of system
) on use RiboGreen
RNA quantification kit (Molecular Probes
) carry out quantitatively.In brief, with TE (10mMTris-HCl pH=7.5,1mM EDTA) dilution 200-doubly with RiboGreen reagent.Generally, 50 μ L reagent are diluted among the 10mL TE.A kind of typical curve of ribosome-RNA(rRNA) is diluted to 2 μ g/mL in TE, then predetermined amount (100,50,40,20,10,5,2 and 0 μ L) ribosome-RNA(rRNA) solution is transferred in new 96 orifice plates (COSTAR#3997), volume is supplied 100 μ L with TE.Generally, with first row's production standard curve of this 96 orifice plate, with the quantitative RNA sample in other hole.Need every part 10 μ L of quantitative RNA sample, they are transferred in the respective aperture of 96 orifice plates, add 90 μ L TE.Each hole adds the RiboGreen reagent of the dilution of a volume (100 μ L), room temperature lucifuge insulation 2-5 minute (10 μ L RNA samples are the 20X dilution in 200 μ L final volume).At STORM
(the Molecular Dynamics of system
) the last fluorescence intensity of measuring each hole.Ribosome-RNA(rRNA) and gained fluorescence intensity creating a standard curve according to known quantity.RNA concentration in the test sample is determined from typical curve, proofreaies and correct according to the 20X dilution.
Reagent and material:
| Product | Company | Catalog number (Cat.No.) | Store |
| DEPC | Sigma | D5758 | 4℃ |
| EDTA | Sigma | E5134 | RT |
| Trizma-Base | Sigma | T8524 | RT |
| Trizma-HCl | Sigma | T7149 | RT |
| Collection Tube Strips | Qiagen | 19562 | RT |
| Ribogreen RNA quantification kit | Molecular Probe | R11490 | -20℃ |
| Rneasy 96 test kits | Qiagen | 74183 | RT |
| Square-Well Blocks | Qiagen | 19573 | RT |
Real-time RT-PCR
Real-time RT-PCR is used TaqMan EZ RT-PCR test kit (Perkin-Elmer Applied Biosystems on ABI Prism 7700 sequence detection systems
) carry out.Utilization is similar to technical literature (Martell etal., 1999.J.Clin.Microbiol.37:327-332) Taqman technology (Roche MolecularDiagnostics Systems) RT-PCR is optimized, make it be suitable for 5 ' IRES of HCV RNA.This system adopts 5 '-3 ' nucleosides lytic activity of AmpliTaq archaeal dna polymerase.In brief, this method is utilized a kind of double-tagging fluorescent hybridization probe (PUTR Probe), it specifically and template between the PCR primer (primer 8125 and 7028) annealing.This probe contains fluorescent reporter molecule (6-Fluoresceincarboxylic acid [FAM]) at 5 ' end, contains fluorescent quenching agent (6-carboxyl tetramethyl-rhodamine [TAMRA]) at its 3 ' end.The emmission spectrum of FAM reporter molecules is suppressed by the quencher on the complete hybridization probe.The nuclease degradation of hybridization probe discharges reporter molecules, causes fluorescent emission to increase.The ABI Prism 7700 sequential detection instrument increases that continuously measured should be launched in the pcr amplification process, thus make the product of amplification and this signal directly proportional.Can represent the analysing amplified collection of illustrative plates of time point (plot) of the logarithmic phase of product accumulation in early days in reaction.The point of having represented the predetermined detection threshold value of the fluorescent signal increase relevant with the exponential growth of PCR product is defined as the cycle threshold (C of this sequential detection instrument
T).C
TValue and HCV RNA input are inversely proportional to; Therefore, under identical PCR condition, the initial concentration of HCV RNA is high more, C
TLow more.ABI Prism 7700 detection systems are passed through C
TTo each standard extent of dilution mapping of known HCV RNA concentration and generate typical curve automatically.
What typical curve was required all is included in each piece RT-PCR flat board with reference to sample.The HCV replicon rna is external synthetic (transcribing by T7), and purifying passes through OD
260Quantitatively.Consider this RNA=2.15X 10 of 1 μ g
11The RNA copy, the preparation diluent is so that have 10
8, 10
7, 10
6, 10
5, 10
4, 10
3Or 10
2Geneome RNA copy/5 μ L.Also mix the cell total rna (50ng/5 μ L) of Huh-7 in each extent of dilution.Every kind of reference standard of 5 μ L (HCV replicon+Huh-7 RNA) is mixed with 45 μ L reagent mixtures, be used for the real-time RT-PCR reaction.
By every kind of cell total rna sample of 5 μ l is mixed with 45 μ L reagent mixtures, set up real-time RT-PCR reaction at the test sample of purifying on the RNeasy 96 hole flat boards.
Reagent and material:
| Product | Company | Catalog number (Cat.No.) | Store |
| TaqMan EZ RT-PCR Kit | PE Applied Biosystems | N808-0236 | -20℃ |
| MicroAmp Optical Caps | PE Applied Biosystems | N801-0935 | RT |
| MicroAmp Optical 96- Well Reaction Plate | PE Applied Biosystems | N801-0560 | RT |
The reagent mix Tetramune:
| Composition | Volume in one duplicate samples (μ L) | Volume (μ L) (91 sample+dead volumes) in flat board | Final concentration |
| The water of no Rnase | 16.5 | 1617 | |
| 5X TaqManEZ damping fluid | 1O | 980 | 1X |
| Mn(OAc) 2(25mM) | 6 | 588 | 3mM |
| dATP(10mM) | 1.5 | 147 | 300μM |
| dCTP(10mM) | 1.5 | 147 | 300μM |
| dGTP(10mM) | 1.5 | 147 | 300μM |
| dUTP(20mM) | 1.5 | 147 | 600μM |
| Forward primer (10 μ M) | 1 | 98 | 200nM |
| Reverse primer (10 μ M) | 1 | 98 | 200nM |
| PUTR probe(5μM) | 2 | 196 | 200nM |
| RTth archaeal dna polymerase (2.5U/ μ L) | 2 | 196 | 0.1U/μL |
| AmpErase UNG (1U/μL) | 0.5 | 49 | 0.01U/μL |
| Cumulative volume | 45 | 4410 |
Forward primer sequence (SEQ ID.2): 5 '-ACG CAG AAA GCG TCT AGC CAT GGCGTT AGT-3 '
Reverse primer sequence (SEQ ID NO.3): 5 '-TCC CGG GGC ACT CGC AAG CAC CCTATC AGG-3 '
Attention: these primer amplifications go out the zone of 256-nt in HCV5 ' non-translational region.
PUTR probe sequence (SEQ ID NO.4):
-TGG TCT GCG GAA CCG GTGAGT ACA CC-
No template contrast (NTC):On each flat board, with 4 holes conducts " NTC ".In these contrasts, every hole adds 5 μ l water and replaces RNA.
Thermal cycle conditions:
50℃ 2min
60℃ 30min
95℃ 5min
After finishing the RT-PCR reaction, data analysis requires to be provided with the fluorescence signal threshold of PCR flat board, by the Ct value is generated typical curve to each with reference to RNA copy number mapping used in the reaction.With the Ct value that derives from test sample interpolation RNA copy number on typical curve.
At last, with the stdn of RNA copy number (quantitative), and be expressed as the genome equivalent/total RNA[ge/ μ of μ g g] based on the RiboGreen RNA that total RNA that each cell cultures hole is extracted carries out.
The RNA copy number in each hole of Tissue Culture Plate [g.e./μ g] is the metric of HCV rna replicon amount under the condition that the different concns inhibitor is arranged.% suppresses in order to equation calculating down:
100-[(g.e./μg inh)/(g.e./μg ctl)x100]。
To be applied to the inhibition concentration data, 50% effective concentration (EC with the nonlinear curve that the Hill pattern is coincide
50) usefulness SAS software (Statistical Software System; SAS Institute, Inc.Cary N.C.) calculates.
In following table 1, use following scope:
IC
50：A＝≥1μM；B＝1μM-500nM；C＜500nM。
Ec
50：A＝≥1μM；and B＝＜1μM
Table 1
Table 2
Sequence table
＜110〉Behringerl. Ying Gehaimu (Canada) limited liability company (Boehringer Ingelheim (Canada) Ltd.)
＜120〉viral polymerase inhibitors
<130>13/095
<140>60/307,674
<141>2001-07-25
<150>60/338,061
<151>2001-12-07
<160>4
<170>FastSEQ for Windows Version 4.0
<210>1
<211>621
<212>PRT
<213>HCV NS5B
<400>1
Met Ser Tyr Tyr His His His His His His Asp Tyr Asp Ile Pro Thr
1 5 10 15
Thr Glu Asn Leu Tyr Phe Gln Gly Ala Met Asp Pro Glu Phe Ser Met
20 25 30
Ser Tyr Thr Trp Thr Gly Ala Leu Ile Thr Pro Cys Ala Ala Glu Glu
35 40 45
Ser Gln Leu Pro Ile Asn Ala Leu Ser Asn Ser Leu Val Arg His Arg
50 55 60
Asn Met Val Tyr Ser Thr Thr Ser Arg Ser Ala Ala Leu Arg Gln Lys
65 70 75 80
Lys Val Thr Phe Asp Arg Leu Gln Val Leu Asp Asp His Tyr Arg Asp
85 90 95
Val Leu Lys Glu Met Lys Ala Lys Ala Ser Thr Val Lys Ala Lys Leu
100 105 110
Leu Ser Val Glu Glu Ala Cys Lys Leu Thr Pro Pro His Ser Ala Lys
115 120 125
Ser Lys Phe Gly Tyr Gly Ala Lys Asp Val Arg Asn Leu Ser Ser Lys
130 135 140
Ala Val Asp His Ile Arg Ser Val Trp Lys Asp Leu Leu Glu Asp Thr
145 150 155 160
Glu Thr Pro Ile Asp Thr Thr Ile Met Ala Lys Asn Glu Val Phe Cys
165 170 175
Val Gln Pro Glu Lys Gly Gly Arg Lys Pro Ala Arg Leu Ile Val Phe
180 185 190
Pro Asp Leu Gly Val Arg Val Cys Glu Lys Met Ala Leu Tyr Asp Val
195 200 205
Val Ser Thr Leu Pro Gln Ala Val Met Gly Ser Ser Tyr Gly Phe Gln
210 215 220
Tyr Ser Pro Lys Gln Arg Val Glu Phe Leu Val Asn Ala Trp Lys Ser
225 230 235 240
Lys Lys Cys Pro Met Gly Phe Ser Tyr Asp Thr Arg Cys Phe Asp Ser
245 250 255
Thr Val Thr Glu Ser Asp Ile Arg Val Glu Glu Ser Ile Tyr Gln Cys
260 265 270
Cys Asp Leu Ala Pro Glu Ala Arg Gln Ala Ile Lys Ser Leu Thr Glu
275 280 285
Arg Leu Tyr Ile Gly Gly Pro Leu Thr Asn Ser Lys Gly Gln Asn Cys
290 295 300
Gly Tyr Arg Arg Cys Arg Ala Ser Gly Val Leu Thr Thr Ser Cys Gly
305 310 315 320
Asn Thr Leu Thr Cys Tyr Leu Lys Ala Ser Ala Ala Cys Arg Ala Ala
325 330 335
Lys Leu Gln Asp Cys Thr Met Leu Val Asn Gly Asp Asp Leu Val Val
340 345 350
Ile Cys Glu Ser Ala Gly Thr Gln Glu Asp Ala Ala Asn Leu Arg Val
355 360 365
Phe Thr Glu Ala Met Thr Arg Tyr Ser Ala Pro Pro Gly Asp Leu Pro
370 375 380
Gln Pro Glu Tyr Asp Leu Glu Leu Ile Thr Ser Cys Ser Ser Asn Val
385 390 395 400
Ser Val Ala His Asp Ala Ser Gly Lys Arg Val Tyr Tyr Leu Thr Arg
405 410 415
Asp Pro Thr Thr Pro Leu Ala Arg Ala Ala Trp Glu Thr Ala Arg His
420 425 430
Thr Pro Ile Asn Ser Trp Leu Gly Asn Ile Ile Met Tyr Ala Pro Thr
435 440 445
Leu Trp Ala Arg Met Val Leu Met Thr His Phe Phe Ser Ile Leu Leu
450 455 460
Ala Gln Glu Gln Leu Glu Lys Ala Leu Asp Cys Gln Ile Tyr Gly Ala
465 470 475 480
Cys Tyr Ser Ile Glu Pro Leu Asp Leu Pro Gln Ile Ile Glu Arg Leu
485 490 495
His Gly Leu Ser Ala Phe Ser Leu His Ser Tyr Ser Pro Gly Glu Ile
500 505 510
Asn Arg Val Ala Ser Cys Leu Arg Lys Leu Gly Val Pro Pro Leu Arg
515 520 525
Val Trp Arg His Arg Ala Arg Ser Val Arg Ala Lys Leu Leu Ser Gln
530 535 540
Gly Gly Arg Ala Ala Thr Cys Gly Lys Tyr Leu Phe Asn Trp Ala Val
545 550 555 560
Arg Thr Lys Leu Lys Leu Thr Pro Ile Pro Ala Ala Ser Arg Leu Asp
565 570 575
Leu Ser Gly Trp Phe Val Ala Gly Tyr Asn Gly Gly Asp Ile Tyr His
580 585 590
Ser Leu Ser Arg Ala Arg Pro Arg Trp Phe Met Leu Cys Leu Leu Leu
595 600 605
Leu Ser Val Gly Val Gly Ile Tyr Leu Leu Pro Asn Arg
610 615 620
<210>2
<211>30
<212>DNA
＜213〉forward primer
<400>2
acgcagaaag cgtctagcca tggcgttagt 30
<210>3
<211>30
<212>DNA
＜213〉reverse primer
<400>3
tcccggggca ctcgcaagca ccctatcagg 30
<210>4
<211>26
<212>DNA
＜213〉PUTR probe
<400>4
tggtctgcgg aaccggtgag tacacc 26
Claims (42)
1. compound and isomer thereof shown in the following formula I, enantiomer, diastereomer, or tautomer:
In the formula:
A is O, S, NR
1, or CR
1, R wherein
1Be selected from: H, (C
1-6) alkyl, it is chosen wantonly and is replaced by following groups:
-halogen, OR
11, SR
11Or N (R
12)
2, R wherein
11And each R
12Be H independently, (C
1-6) alkyl, (C
3-7) cycloalkyl, (C
1-6) alkyl-(C
3-7) cycloalkyl, (C
1-6) alkyl-aryl or (C
1-6) alkyl-Het, described aryl or Het are optional by R
10Replace; Perhaps
Two R
12, and be attached on the nitrogen that they connect by covalent bonds together, form 5,6 or the saturated heterocycle of 7-person;
-----represent singly-bound or two key;
R
2Be selected from: H, halogen, R
21, OR
21, SR
21, COOR
21, SO
2N (R
22)
2, N (R
22)
2, CON (R
22)
2, NR
22C (O) R
22Or NR
22C (O) NR
22, R wherein
21And each R
22Be H independently, (C
1-6) alkyl, haloalkyl, (C
2-6) alkenyl, (C
3-7) cycloalkyl, (C
2-6) alkynyl, (C
5-7) cycloalkenyl group, 6 or 10-person's aryl or Het, described R
21And R
22Optional by R
20Replace perhaps two R
22Be combined together to form 5,6 or the saturated heterocycle of 7-person with its nitrogen that is connected;
R wherein
10And R
20Respectively do for oneself:
-1～4 substituting groups are selected from: halogen, OPO
3H, NO
2, cyano group, azido-, C (=NH) NH
2, C (=NH) NH (C
1-6) alkyl or C (=NH) NHCO (C
1-6) alkyl; Perhaps
-1～4 substituting groups are selected from:
A) (C
1-6) alkyl or haloalkyl, (C
3-7) cycloalkyl, optional comprise 1 or 2 heteroatomic C
3-7Spiro cycloalkyl group, (C
2-6) alkenyl, (C
3-6) cycloalkenyl group, (C
2-8) alkynyl, (C
1-6) alkyl-(C
3-7) cycloalkyl, all these groups are all randomly by R
150Replace;
B) OR
104, R wherein
104Be H, (C
1-6Alkyl), (C
3-7) cycloalkyl, or (C
1-6) alkyl-(C
3-7) cycloalkyl, aryl, Het, (C
1-6Alkyl) aryl or (C
1-6Alkyl) Het, described alkyl, cycloalkyl, aryl, Het, (C
1-6Alkyl) aryl or (C
1-6Alkyl) Het is all randomly by R
150Replace;
C) OCOR
105, R wherein
105Be (C
1-6) alkyl, (C
3-7) cycloalkyl, (C
1-6) alkyl-(C
3-7) cycloalkyl, Het, (C
1-6Alkyl) aryl or (C
1-6Alkyl) Het, described alkyl, cycloalkyl, aryl, Het, (C
1-6Alkyl) aryl or (C
1-6Alkyl) Het is all randomly by R
150Replace;
D) SR
108, SO
2N (R
108)
2Or SO
2N (R
108) C (O) R
108, each R wherein
108Be H independently, (C
1-6) alkyl, (C
3-7) cycloalkyl or (C
1-6) alkyl-(C
3-7) cycloalkyl, aryl, Het, (C
1-6Alkyl) aryl or (C
1-6Alkyl) Het, perhaps two R
108Combine formation 5,6 or the saturated heterocycle of 7-person, described alkyl, cycloalkyl, aryl, Het, (C by the connected nitrogen of covalent linkage
1-6Alkyl) aryl or (C
1-6Alkyl) Het or heterocycle are all optional by R
150Replace;
E) NR
111R
112, R wherein
111Be H, (C
1-6) alkyl, (C
3-7) cycloalkyl or (C
1-6) alkyl-(C
3-7) cycloalkyl, aryl, Het, (C
1-6Alkyl) aryl or (C
1-6Alkyl) Het, and R
112Be H, CN, (C
1-6) alkyl, (C
3-7) cycloalkyl or (C
1-6) alkyl-(C
3-7) cycloalkyl, aryl, Het, (C
1-6Alkyl) aryl, (C
1-6Alkyl) Het, COOR
115Or SO
2R
115, R wherein
115Be (C
1-6) alkyl, (C
3-7) cycloalkyl, or (C
1-6) alkyl-(C
3-7) cycloalkyl, aryl, Het, (C
1-6Alkyl) aryl or (C
1-6Alkyl) Het, perhaps two R
111And R
112Be combined together to form 5,6 or the saturated heterocycle of 7-person by the connected nitrogen of covalent linkage, described alkyl, cycloalkyl, aryl, Het, (C
1-6Alkyl) aryl or (C
1-6Alkyl) Het, or heterocycle is all randomly by R
150Replace;
F) NR
116COR
117, R wherein
116And R
117The H that respectively does for oneself, (C
1-6) alkyl, (C
3-7) cycloalkyl, (C
1-6) alkyl-(C
3-7) cycloalkyl, aryl, Het, (C
1-6Alkyl) aryl or (C
1-6Alkyl) Het, described (C
1-6) alkyl, (C
3-7) cycloalkyl, (C
1-6) alkyl-(C
3-7) cycloalkyl, aryl, Het, (C
1-6Alkyl) aryl or (C
1-6Alkyl) Het is all randomly by R
150Replace;
G) NR
118CONR
119R
120, R wherein
118, R
119And R
120The H that respectively does for oneself, (C
1-6) alkyl, (C
3-7) cycloalkyl, (C
1-6) alkyl-(C
3-7) cycloalkyl, aryl, Het, (C
1-6Alkyl) aryl or (C
1-6Alkyl) Het, perhaps R
118By covalent linkage and R
119In conjunction with and the nitrogen that is connected with them combine, form 5,6 or the saturated heterocycle of 7-person;
Perhaps R
119And R
120The nitrogen coupled by covalent linkage combines, formation 5,6 or the saturated heterocycle of 7-person, described alkyl, cycloalkyl, (C
1-6) alkyl-(C
3-7) cycloalkyl, aryl, Het, (C
1-6Alkyl) aryl or (C
1-6Alkyl) Het or heterocycle are all randomly by R
150Replace;
H) NR
121COCOR
122, R wherein
121And R
122The H that respectively does for oneself, (C
1-6) alkyl, (C
3-7) cycloalkyl, (C
1-6) alkyl-(C
3-7) cycloalkyl, 6-or 10-person's aryl, Het, (C
1-6Alkyl) aryl or (C
1-6Alkyl) Het, described alkyl, cycloalkyl, alkyl-cycloalkyl, aryl, Het, (C
1-6Alkyl) aryl or (C
1-6Alkyl) Het is all randomly by R
150Replace;
Perhaps R
122Be OR
123Or N (R
124)
2, R wherein
123And each R
124Be H independently, (C
1-6Alkyl), (C
3-7) cycloalkyl, (C
1-6) alkyl-(C
3-7) cycloalkyl, aryl, Het, (C
1-6Alkyl) aryl or (C
1-6Alkyl) Het, perhaps R
124Be OH or O (C
1-6Alkyl), perhaps two R
124By covalent bonds together, formation 5,6 or the saturated heterocycle of 7-person, described alkyl, cycloalkyl, alkyl-cycloalkyl, aryl, Het, (C
1-6Alkyl) aryl or (C
1-6Alkyl) Het and heterocycle are all randomly by R
150Replace;
I) COR
127, R wherein
127Be H, (C
1-6) alkyl, (C
3-7) cycloalkyl or (C
1-6) alkyl-(C
3-7) cycloalkyl, aryl, Het, (C
1-6Alkyl) aryl or (C
1-6Alkyl) Het, described alkyl, cycloalkyl, aryl, Het, (C
1-6Alkyl) aryl or (C
1-6Alkyl) Het is all randomly by R
150Replace;
J) COOR
128R wherein
128Be H, (C
1-6) alkyl, (C
3-7) cycloalkyl, (C
1-6) alkyl-(C
3-7) cycloalkyl, aryl, Het, (C
1-6Alkyl) aryl or (C
1-6Alkyl) Het, described (C
1-6) alkyl, (C
3-7) cycloalkyl, or (C
1-6) alkyl-(C
3-7) cycloalkyl, aryl, Het, (C
1-6Alkyl) aryl and (C
1-6Alkyl) Het is all randomly by R
150Replace;
K) CONR
129R
130, R wherein
129And R
130Be H independently, (C
1-6) alkyl, (C
3-7) cycloalkyl, (C
1-6) alkyl-(C
3-7) cycloalkyl, aryl, Het, (C
1-6Alkyl) aryl or (C
1-6Alkyl) Het, perhaps two R
129And R
130The nitrogen coupled by covalent linkage combines, formation 5,6 or the saturated heterocycle of 7-person, described alkyl, cycloalkyl, alkyl-cycloalkyl, aryl, Het, (C
1-6Alkyl) aryl, (C
1-6Alkyl) Het and heterocycle are all randomly by R
150Replace;
L) aryl, Het, (C
1-6Alkyl) aryl or (C
1-6Alkyl) Het, all these groups are all randomly by R
150Replace; R wherein
150Be defined as follows:
-1～3 substituting groups are selected from: halogen, OPO
3H, NO
2, cyano group, azido-, C (=NH) NH
2, C (=NH) NH (C
1-6) alkyl or C (=NH) NHCO (C
1-6) alkyl; Perhaps
-1～3 substituting groups are selected from:
A) (C
1-6) alkyl or haloalkyl, (C
3-7) cycloalkyl, optional comprise 1 or 2 heteroatomic C
3-7Spiro cycloalkyl group, (C
2-6) alkenyl, (C
2-8) alkynyl, (C
1-6) alkyl-(C
3-7) cycloalkyl, all these groups are all optional by R
160Replace;
B) OR
104, R wherein
104Be H, (C
1-6Alkyl), (C
3-7) cycloalkyl, (C
1-6) alkyl-(C
3-7) cycloalkyl, aryl, Het, (C
1-6Alkyl) aryl or (C
1-6Alkyl) Het, described alkyl, cycloalkyl, aryl, Het, (C
1-6Alkyl) aryl or (C
1-6Alkyl) Het is all optional by R
160Replace;
C) OCOR
105, R wherein
105Be (C
1-6) alkyl, (C
3-7) cycloalkyl, (C
1-6) alkyl-(C
3-7) cycloalkyl, Het, (C
1-6Alkyl) aryl or (C
1-6Alkyl) Het, described alkyl, cycloalkyl, aryl, Het, (C
1-6Alkyl) aryl or (C
1-6Alkyl) Het is all optional by R
160Replace;
D) SR
108, SO
2N (R
108)
2Or SO
2N (R
108) C (O) R
108, each R wherein
108Be H independently, (C
1-6) alkyl, (C
3-7) cycloalkyl or (C
1-6) alkyl-(C
3-7) cycloalkyl, aryl, Het, (C
1-6Alkyl) aryl or (C
1-6Alkyl) Het, perhaps two R
108The nitrogen coupled by covalent linkage combines, formation 5,6 or the saturated heterocycle of 7-person, described alkyl, cycloalkyl, aryl, Het, (C
1-6Alkyl) aryl or (C
1-6Alkyl) Het or heterocycle are all optional by R
160Replace;
E) NR
111R
112, R wherein
111Be H, (C
1-6) alkyl, (C
3-7) cycloalkyl or (C
1-6) alkyl-(C
3-7) cycloalkyl, aryl, Het, (C
1-6Alkyl) aryl or (C
1-6Alkyl) Het, and R
112Be H, CN, (C
1-6) alkyl, (C
3-7) cycloalkyl or (C
1-6) alkyl-(C
3-7) cycloalkyl, aryl, Het, (C
1-6Alkyl) aryl, (C
1-6Alkyl) Het, COOR
115Or SO
2R
115, R wherein
115Be (C
1-6) alkyl, (C
3-7) cycloalkyl, (C
1-6) alkyl-(C
3-7) cycloalkyl, aryl, Het, (C
1-6Alkyl) aryl or (C
1-6Alkyl) Het, perhaps two R
111And R
112The nitrogen coupled by covalent linkage combines, formation 5,6 or the saturated heterocycle of 7-person, described alkyl, cycloalkyl, aryl, Het, (C
1-6Alkyl) aryl or (C
1-6Alkyl) Het, or heterocycle is all optional by R
160Replace;
F) NR
116COR
117, R wherein
116And R
117The H that respectively does for oneself, (C
1-6) alkyl, (C
3-7) cycloalkyl, (C
1-6) alkyl-(C
3-7) cycloalkyl, aryl, Het, (C
1-6Alkyl) aryl or (C
1-6Alkyl) Het, described (C
1-6) alkyl, (C
3-7) cycloalkyl, (C
1-6) alkyl-(C
3-7) cycloalkyl, aryl, Het, (C
1-6Alkyl) aryl or (C
1-6Alkyl) Het is all optional by R
160Replace;
G) NR
118CONR
119R
120, R wherein
118, R
119And R
120The H that respectively does for oneself, (C
1-6) alkyl, (C
3-7) cycloalkyl, (C
1-6) alkyl-(C
3-7) cycloalkyl, aryl, Het, (C
1-6Alkyl) aryl or (C
1-6Alkyl) Het, perhaps R
118By covalent linkage and R
119In conjunction with and the nitrogen combination that is connected with it, form 5,6 or the saturated heterocycle of 7-person, perhaps R
119And R
120The nitrogen coupled by covalent linkage combines, formation 5,6 or the saturated heterocycle of 7-person, described alkyl, cycloalkyl, (C
1-6) alkyl-(C
3-7) cycloalkyl, aryl, Het, (C
1-6Alkyl) aryl or (C
1-6Alkyl) Het or heterocycle are all optional by R
160Replace;
H) NR
121COCOR
122, R wherein
121And R
122The H that respectively does for oneself, (C
1-6) alkyl, (C
3-7) cycloalkyl, (C
1-6) alkyl-(C
3-7) cycloalkyl, 6-or 10-person's aryl, Het, (C
1-6Alkyl) aryl or (C
1-6Alkyl) Het, described alkyl, cycloalkyl, alkyl-cycloalkyl, aryl, Het, (C
1-6Alkyl) aryl or (C
1-6Alkyl) Het is all optional by R
160Replace perhaps R
122Be OR
123Or N (R
124)
2, R wherein
123And each R
124Be H independently, (C
1-6Alkyl), (C
3-7) cycloalkyl, (C
1-6) alkyl-(C
3-7) cycloalkyl, aryl, Het, (C
1-6Alkyl) aryl or (C
1-6Alkyl) Het, perhaps R
124Be OH or O (C
1-6Alkyl), perhaps two R
124By covalent bonds together, formation 5,6 or the saturated heterocycle of 7-person, described alkyl, cycloalkyl, alkyl-cycloalkyl, aryl, Het, (C
1-6Alkyl) aryl or (C
1-6Alkyl) Het and heterocycle are all optional by R
160Replace;
I) COR
127, R wherein
127Be H, (C
1-6) alkyl, (C
3-7) cycloalkyl or (C
1-6) alkyl-(C
3-7) cycloalkyl, aryl, Het, (C
1-6Alkyl) aryl or (C
1-6Alkyl) Het, described alkyl, cycloalkyl, aryl, Het, (C
1-6Alkyl) aryl or (C
1-6Alkyl) Het is all optional by R
160Replace;
J) tetrazolium, COOR
128, R wherein
128Be H, (C
1-6) alkyl, (C
3-7) cycloalkyl, (C
1-6) alkyl-(C
3-7) cycloalkyl, aryl, Het, (C
1-6Alkyl) aryl or (C
1-6Alkyl) Het, described (C
1-6) alkyl, (C
3-7) cycloalkyl, (C
1-6) alkyl-(C
3-7) cycloalkyl, aryl, Het, (C
1-6Alkyl) aryl and (C
1-6Alkyl) Het is all optional by R
160Replace; And
K) CONR
129R
130, R wherein
129And R
130Be H independently, (C
1-6) alkyl, (C
3-7) cycloalkyl, (C
1-6) alkyl-(C
3-7) cycloalkyl, aryl, Het, (C
1-6Alkyl) aryl or (C
1-6Alkyl) Het, perhaps two R
129And R
130The nitrogen coupled by covalent linkage combines, formation 5,6 or the saturated heterocycle of 7-person, described alkyl, cycloalkyl, alkyl-cycloalkyl, aryl, Het, (C
1-6Alkyl) aryl, (C
1-6Alkyl) Het and heterocycle are all optional by R
160Replace;
R wherein
160Be defined as 1 or 2 substituting group, be selected from: tetrazolium, halogen, CN, C
1-6Alkyl, haloalkyl, COOR
161, SO
3H, SR
161, SO
2R
161, OR
161, N (R
162)
2, SO
2N (R
162)
2, NR
162COR
162Or CON (R
162)
2, R here
161And each R
162Be H independently, (C
1-6) alkyl, (C
3-7) cycloalkyl or (C
1-6) alkyl-(C
3-7) cycloalkyl; Perhaps two R
162The nitrogen coupled by covalent linkage combines, formation 5,6 or the saturated heterocycle of 7-person;
B is NR
3Or CR
3, condition is that one of A or B are CR
1Or CR
3, R wherein
3Be selected from: (C
1-6) alkyl, haloalkyl, (C
3-7) cycloalkyl, (C
5-7) cycloalkenyl group, (C
6-10) bicyclic alkyl, (C
6-10) bicycloenyl, 6-or 10-person's aryl, Het, (C
1-6) alkyl-aryl or (C
1-6) alkyl-Het,
Described alkyl, cycloalkyl, bicyclic alkyl, aryl, Het, alkyl-aryl and alkyl-Het be optional to be selected from following substituting group by 1～4 and to replace: halogen, perhaps
A) (C
1-6) alkyl, it is chosen wantonly and is replaced by following groups:
-OR
31Or SR
31, R wherein
31Be H, (C
1-6Alkyl), (C
3-7) cycloalkyl, (C
1-6) alkyl-(C
3-7) cycloalkyl, aryl, Het, (C
1-6) alkyl-aryl or (C
1-6) alkyl-Het; Perhaps
-N (R
32)
2, each R wherein
32Be H independently, (C
1-6) alkyl, (C
3-7) cycloalkyl, (C
1-6) alkyl-(C
3-7) cycloalkyl, aryl, Het, (C
1-6) alkyl-aryl or (C
1-6) alkyl-Het; Perhaps two R
32The nitrogen coupled by covalent linkage combines, formation 5,6 or the saturated heterocycle of 7-person;
B) OR
33, R wherein
33Be H, (C
1-6) alkyl, (C
3-7) cycloalkyl or (C
1-6) alkyl-(C
3-7) cycloalkyl, aryl, Het, (C
1-6) alkyl-aryl or (C
1-6) alkyl-Het;
C) SR
34, R wherein
34Be H, (C
1-6) alkyl, (C
3-7) cycloalkyl, perhaps (C
1-6) alkyl-(C
3-7) cycloalkyl, aryl, Het, (C
1-6) alkyl-aryl or (C
1-6) alkyl-Het;
And
D) N (R
35)
2, each R wherein
35Be H independently, (C
1-6) alkyl, (C
3-7) cycloalkyl, (C
1-6) alkyl-(C
3-7) cycloalkyl, aryl, Het, (C
1-6) alkyl-aryl or (C
1-6) alkyl-Het; Perhaps two R
35The nitrogen coupled by covalent linkage combines, formation 5,6 or the saturated heterocycle of 7-person;
K is N or CR
4, R wherein
4Be H, halogen, (C
1-6) alkyl, haloalkyl, (C
3-7) cycloalkyl or (C
1-6) alkyl-(C
3-7) cycloalkyl; Perhaps R
4Be OR
41Or SR
41, COR
41Or NR
41COR
41, each R wherein
41Be H independently, (C
1-6) alkyl), (C
3-7) cycloalkyl or (C
1-6) alkyl-(C
3-7) cycloalkyl, perhaps R
4Be NR
42R
43, R wherein
42And R
43Be H independently, (C
1-6) alkyl, (C
3-7) cycloalkyl, (C
1-6) alkyl-(C
3-7) cycloalkyl, perhaps two R
42And R
43The nitrogen coupled by covalent linkage combines, formation 5,6 or the saturated heterocycle of 7-person;
L is N or CR
5, R wherein
5Definition and above-mentioned R
4Definition identical;
M is N or CR
7, R wherein
7Definition and above-mentioned R
4Definition identical;
Y
1Be O or S;
Z is OR
6, R wherein
6Be H, (C
1-6) alkyl, it is chosen wantonly and is replaced by following groups: halogen, hydroxyl, carboxyl, amino, C
1-6Alkoxyl group, C
1-6Alkoxy carbonyl, and C
1-6Alkylamino; Perhaps R
6Be C
1-6Alkylaryl, it is chosen wantonly and is replaced by following groups: halogen, cyano group, nitro, C
1-6Alkyl, C
1-6Haloalkyl, C
1-6Alkyloyl ,-(CH
2)
1-6-COOR
7,-(CH
2)
1-6-CONR
7R
8,-(CH
2)
1-6-NR
7R
8,-(CH
2)
1-6-NR
7COR
8,-(CH
2)
1-6-NHSO
2R
7,-(CH
2)
1-6-OR
7,-(CH
2)
1-6-SR
7,-(CH
2)
1-6-SO
2R
7, and-(CH
2)
1-6-SO
2NR
7R
8, each R wherein
7And each R
8Be H or C
1-6Alkyl,
Perhaps Z is NR
9R
10, each R wherein
9And R
10Be selected from: H, C
1-6Alkoxyl group, or C
1-6Alkyl, it is optional by halogen, hydroxyl, carboxyl, amino, C
1-6Alkoxyl group, C
1-6Alkoxy carbonyl, and C
1-6Alkylamino replaces;
Or its salt;
Condition is, when A is CR
1, R
1Be Me, R
2For pyridine or
B is NR
3, R
3Be Me, K, L, M are CH, Y
1Be O, and Z is OR
6The time, R
6Be not H; The condition of reaching is, when A is NR
1, R
1Be H, R
2Be phenyl, B is CR
3, R
3Be phenyl, K, L, M are CH, Y
1Be O, and Z is OR
6The time, R
6Be not H;
And condition is, when A is S, and R
2Be bromine, B is CR
3, R
3Be Me, K is CH, and L is CH, and M is CR
7, R
7Be H or Me, Y
1Be O, and Z is OR
6The time, R
6Be not H;
And condition is, when A is O, and R
2Be H, B is CR
3, RA
3Be phenyl, K, L, M are CH, Y
1Be O, and Z is OR
6The time, R
6Be not H;
The condition of reaching is, when A is CR
1, R
1Be Me, R
2Be pyridine, B is NR
3, R
3Be Me, K, L, M are CH, Y
1Be O, and Z is OR
6The time, R
6Be not Me;
The condition of reaching is, when A is CR
1, R
1Be Me, R
2For
B is NR
3, R
3Be Me, K, L, M are CH, Y
1Be O, and Z is OR
6The time, R
6Be not Et;
The condition of reaching is, when A is CR
1, R
1Be CH, R
2Be Me, B is NR
3, R
3Be Me, K, L, M are CH, Y
1Be O, and Z is OR
6The time R
6Be not Et;
The condition of reaching is, when A is CR
1, R
1Be Et, R
2Be Me, B is NR
3, R
3Be Me, K, L, M are CH, Y
1Be O, and Z is OR
6The time, R
6Be not CH
2CH
2N (Me)
2
And condition is, when A is CH, and R
2Be Me, B is NR
3, R
3For
K is N, and L is CR
5, R
5Be Me, M is CR
7, R
7Be OH, Y
1Be O, and Z is OR
6The time, R
6Be not Et;
The condition of reaching is, when A is NR
1, R
1Be Me, R
2Be Br, B is CR
3, R
3For
K is N, and L is CR
5, R
5Be Me, M is CR
7, R
7Be Br, Y
1Be O, and Z is OR
6The time, R
6Be not Me;
The condition of reaching is, when A is NR
1, R
1Be H, R
2Be Cl, B is CR
3, R
3Be Et, K is CH, and L is CH, and M is CH, Y
1Be O, Z is OR
6The time, R
6Be not Me;
The condition of reaching is, when A is NR
1, R
1Be H, R
2Be phenyl, B is CR
3, R
3Be phenyl, K is CH, and L is CH, and M is CR
7, R
7Be Me, Y
1Be O, Z is OR
6The time, R
6Be not Et;
The condition of reaching is, when A is NR
1, R
1Be H, R
2For
B is CR
3, R
3For
K is CH, and L is N, and M is CH, Y
1Be O, and Z is OR
6The time, R
6Be not Et;
And condition is, when A is S, and R
2Be Br, B is CR
3, R
3Be Me, K is CH, and L is CH, and M is CH, Y
1Be O, and Z is OR
6The time, R
6Be not Me;
The condition of reaching is, when A is NR
1, R
1Be H, R
2For
B is NR
3, R
3Be cyclohexyl, K, L, M are CH, Y
1Be O, Z is OR
6The time, R
6Be not H.
2. according to the compound of claim 1, it has the structure of following formula (II):
In the formula, preferred A is O, S, or NR
1
3. according to the compound of claim 2, wherein A is NR
1
4. according to the compound of claim 1, it has the structure shown in the following formula (III):
In the formula, preferred B is NR
3
5. according to the compound of claim 1, M wherein, K and L are CH or N.
6. according to the compound of claim 5, M wherein, K and L are CH.
7. according to the compound of claim 1, it has following structure shown in various:
Perhaps
R in the formula
1, R
2, R
3With Z as defined in claim 1.
8. according to the compound of claim 1, R wherein
1Be selected from: H or (C
1-6) alkyl.
9. compound according to Claim 8, wherein R
1Be H, CH
3, sec.-propyl, or isobutyl-.
10. according to the compound of claim 9, R wherein
1Be H or CH
3
11. according to the compound of claim 10, wherein R
1Be CH
3
12. according to the compound of claim 1, wherein R
2Be selected from: H, halogen, (C
2-6) alkenyl, (C
5-7) cycloalkenyl group, 6-or 10-person's aryl or Het; (C wherein
2-6) alkenyl, (C
5-7) cycloalkenyl group, aryl or Het are optional by R
20Replace R wherein
20Be defined as:
-1～4 substituting groups are selected from: halogen, NO
2, cyano group, azido-, C (=NH) NH
2, C (=NH) NH (C
1-6) alkyl or C (=NH) NHCO (C
1-6) alkyl; Perhaps
-1～4 substituting groups are selected from:
A) (C
1-6) alkyl or haloalkyl, (C
3-7) cycloalkyl, (C
2-6) alkenyl, (C
2-8) alkynyl, (C
1-6) alkyl-(C
3-7) cycloalkyl, all these groups are optional by R
150Replace;
B) OR
104, R wherein
104Be H, (C
1-6Alkyl), (C
3-7) cycloalkyl, or (C
1-6) alkyl-(C
3-7) cycloalkyl, aryl, Het, (C
1-6Alkyl) aryl or (C
1-6Alkyl) Het, described alkyl, cycloalkyl, aryl, Het, (C
1-6Alkyl) aryl or (C
1-6Alkyl) Het is all randomly by R
150Replace;
C) OCOR
105, R wherein
105Be (C
1-6) alkyl, (C
3-7) cycloalkyl, (C
1-6) alkyl-(C
3-7) cycloalkyl, Het, (C
1-6Alkyl) aryl or (C
1-6Alkyl) Het, described alkyl, cycloalkyl, aryl, Het, (C
1-6Alkyl) aryl or (C
1-6Alkyl) Het is all randomly by R
150Replace;
D) SR
108, SO
2N (R
108)
2Or SO
2N (R
108) C (O) R
108, each R wherein
108Be H independently, (C
1-6) alkyl, (C
3-7) cycloalkyl or (C
1-6) alkyl-(C
3-7) cycloalkyl, aryl, Het, (C
1-6Alkyl) aryl or (C
1-6Alkyl) Het, perhaps two R
108The nitrogen coupled by covalent linkage combines, formation 5,6 or the saturated heterocycle of 7-person, described alkyl, cycloalkyl, aryl, Het, (C
1-6Alkyl) aryl or (C
1-6Alkyl) Het or heterocycle are all randomly by R
150Replace;
E) NR
111R
112, R wherein
111Be H, (C
1-6) alkyl, (C
3-7) cycloalkyl or (C
1-6) alkyl-(C
3-7) cycloalkyl, aryl, Het, (C
1-6Alkyl) aryl or (C
1-6Alkyl) Het, and R
112Be H, CN, (C
1-6) alkyl, (C
3-7) cycloalkyl or (C
1-6) alkyl-(C
3-7) cycloalkyl, aryl, Het, (C
1-6Alkyl) aryl, (C
1-6Alkyl) Het, COOR
115Or SO
2R
115, R wherein
115Be (C
1-6) alkyl, (C
3-7) cycloalkyl, or (C
1-6) alkyl-(C
3-7) cycloalkyl, aryl, Het, (C
1-6Alkyl) aryl or (C
1-6Alkyl) Het, perhaps two R
111And R
112The nitrogen coupled by covalent linkage combines, formation 5,6 or the saturated heterocycle of 7-person, described alkyl, cycloalkyl, aryl, Het, (C
1-6Alkyl) aryl or (C
1-6Alkyl) Het, or heterocycle is all randomly by R
150Replace;
F) NR
116COR
117, R wherein
116And R
117The H that respectively does for oneself, (C
1-6) alkyl, (C
3-7) cycloalkyl, (C
1-6) alkyl-(C
3-7) cycloalkyl, aryl, Het, (C
1-6Alkyl) aryl or (C
1-6Alkyl) Het, described (C
1-6) alkyl, (C
3-7) cycloalkyl, (C
1-6) alkyl-(C
3-7) cycloalkyl, aryl, Het, (C
1-6Alkyl) aryl or (C
1-6Alkyl) Het is all randomly by R
150Replace;
G) NR
118CONR
119R
120, R wherein
118, R
119And R
120The H that respectively does for oneself, (C
1-6) alkyl, (C
3-7) cycloalkyl, (C
1-6) alkyl-(C
3-7) cycloalkyl, aryl, Het, (C
1-6Alkyl) aryl or (C
1-6Alkyl) Het, perhaps R
118By covalent linkage and R
119In conjunction with and the nitrogen that is connected with them combine, form 5,6 or the saturated heterocycle of 7-person;
Perhaps R
119And R
120The nitrogen coupled by covalent linkage combines, formation 5,6 or the saturated heterocycle of 7-person;
Described alkyl, cycloalkyl, (C
1-6) alkyl-(C
3-7) cycloalkyl, aryl, Het, (C
1-6Alkyl) aryl or (C
1-6Alkyl) Het or heterocycle are all randomly by R
150Replace;
H) NR
121COCOR
122, R wherein
121And R
122The H that respectively does for oneself, (C
1-6) alkyl, (C
3-7) cycloalkyl, (C
1-6) alkyl-(C
3-7) cycloalkyl, 6-or 10-person's aryl, Het, (C
1-6Alkyl) aryl or (C
1-6Alkyl) Het, described alkyl, cycloalkyl, alkyl-cycloalkyl, aryl, Het, (C
1-6Alkyl) aryl or (C
1-6Alkyl) Het is all randomly by R
150Replace;
Perhaps R
122Be OR
123Or N (R
124)
2, R wherein
123And each R
124Be H independently, (C
1-6Alkyl), (C
3-7) cycloalkyl, or (C
1-6) alkyl-(C
3-7) cycloalkyl, aryl, Het, (C
1-6Alkyl) aryl or (C
1-6Alkyl) Het, perhaps R
124Be OH or O (C
1-6Alkyl), perhaps two R
124By covalent bonds together, formation 5,6 or the saturated heterocycle of 7-person, described alkyl, cycloalkyl, alkyl-cycloalkyl, aryl, Het, (C
1-6Alkyl) aryl or (C
1-6Alkyl) Het and heterocycle are all randomly by R
150Replace;
I) COR
127, R wherein
127Be H, (C
1-6) alkyl, (C
3-7) cycloalkyl or (C
1-6) alkyl-(C
3-7) cycloalkyl, aryl, Het, (C
1-6Alkyl) aryl or (C
1-6Alkyl) Het, described alkyl, cycloalkyl, aryl, Het, (C
1-6Alkyl) aryl or (C
1-6Alkyl) Het is all randomly by R
150Replace;
J) COOR
128, R wherein
128Be H, (C
1-6) alkyl, (C
3-7) cycloalkyl, or (C
1-6) alkyl-(C
3-7) cycloalkyl, aryl, Het, (C
1-6Alkyl) aryl or (C
1-6Alkyl) Het, described (C
1-6) alkyl, (C
3-7) cycloalkyl, or (C
1-6) alkyl-(C
3-7) cycloalkyl, aryl, Het, (C
1-6Alkyl) aryl and (C
1-6Alkyl) Het is all randomly by R
150Replace;
K) CONR
129R
130, R wherein
129And R
130Be H independently, (C
1-6) alkyl, (C
3-7) cycloalkyl, (C
1-6) alkyl-(C
3-7) cycloalkyl, aryl, Het, (C
1-6Alkyl) aryl or (C
1-6Alkyl) Het, perhaps two R
129And R
130The nitrogen coupled by covalent linkage combines, formation 5,6 or the saturated heterocycle of 7-person, described alkyl, cycloalkyl, alkyl-cycloalkyl, aryl, Het, (C
1-6Alkyl) aryl, (C
1-6Alkyl) Het and heterocycle are all randomly by R
150Replace;
L) aryl, Het, (C1-6 alkyl) aryl or (C1-6 alkyl) Het, all these groups are all optional by R
150Replace R wherein
150Be preferably:
-1～3 substituting groups, it is selected from: halogen, NO
2, cyano group or azido-; Perhaps
-1～3 substituting groups, it is selected from:
A) (C
1-6) alkyl or haloalkyl, (C
3-7) cycloalkyl, (C
2-6) alkenyl, (C
2-8) alkynyl, (C
1-6) alkyl-(C
3-7) cycloalkyl, all these groups are all randomly by R
160Replace;
B) ORA
104, R wherein
104Be H, (C
1-6Alkyl) or (C
3-7) cycloalkyl, described alkyl or cycloalkyl is all randomly by R
160Replace;
D) SR
108, SO
2N (R
108)
2Or SO
2N (R
108) C (O) R
108, each R wherein
108All be H independently, (C
1-6) alkyl, (C
3-7) cycloalkyl or (C
1-6) alkyl-(C
3-7) cycloalkyl, aryl, Het, perhaps two R
108The nitrogen coupled by covalent linkage combines, formation 5,6 or the saturated heterocycle of 7-person, and described alkyl, cycloalkyl, aryl, Het and heterocycle are all optional by R
160Replace;
E) NR
111R
112, R wherein
111Be H, (C
1-6) alkyl, or (C
3-7) cycloalkyl, and R
112Be H, (C
1-6) alkyl or (C
3-7) cycloalkyl, COOR
115Or SO
2R
115, R wherein
115Be (C
1-6) alkyl or (C
3-7) cycloalkyl, perhaps two R
111And R
112The nitrogen coupled by covalent linkage combines, and formation 5,6 or the saturated heterocycle of 7-person, described alkyl, cycloalkyl and heterocycle are all optional by R
160Replace;
F) NR
116COR
117, R wherein
116And R
117The H that respectively does for oneself, (C
1-6) alkyl or (C
3-7) described (C of cycloalkyl
1-6) alkyl and (C
3-7) cycloalkyl is all optional by R
160Replace;
G) NR
118CONR
119R
120, R wherein
118, R
119And R
120The H that respectively does for oneself, (C
1-6) alkyl or (C
3-7) cycloalkyl, perhaps R
118By covalent linkage and R
119In conjunction with and the nitrogen that is connected with them combine, form 5,6 or the saturated heterocycle of 7-person, perhaps R
119And R
120The nitrogen coupled by covalent linkage combines, formation 5,6 or the saturated heterocycle of 7-person, and described alkyl, cycloalkyl, and heterocycle is all optional by R
160Replace;
H) NR
121COCOR
122, R wherein
121Be H, (C
1-6) alkyl or (C
3-7) cycloalkyl, described alkyl and cycloalkyl are all optional by R
160Replace perhaps R
122Be OR
123Or N (R
124)
2, R wherein
123And each R
124Be H independently, (C
1-6Alkyl) or (C
3-7) cycloalkyl, perhaps two R
124By covalent bonds together, formation 5,6 or the saturated heterocycle of 7-person, described alkyl, cycloalkyl and heterocycle are all optional by R
160Replace;
I) COR
127, R wherein
127Be H, (C
1-6) alkyl or (C
3-7) cycloalkyl, described alkyl and cycloalkyl are all optional by R
160Replace;
J) COOR
128, R wherein
128Be H, (C
1-6) alkyl or (C
3-7) cycloalkyl, described (C
1-6) alkyl and (C
3-7) cycloalkyl is all optional by R
160Replace; And
K) CONR
129R
130, R wherein
129And R
130Be H independently, (C
1-6) alkyl or (C
3-7) cycloalkyl, perhaps two R
129And R
130The nitrogen coupled by covalent linkage combines, and formation 5,6 or the saturated heterocycle of 7-person, described alkyl, cycloalkyl and heterocycle are all optional by R
160Replace;
R wherein
160Be defined as 1 or 2 and be selected from following substituting group:
Halogen, CN, C
1-6Alkyl, haloalkyl, COOR
161, OR
161, N (R
162)
2, SO
2N (R
162)
2, NR
162COR
162Or CON (R
162)
2, R wherein
161And each R
162Be H independently, (C
1-6) alkyl, (C
3-7) cycloalkyl or (C
1-6) alkyl-(C
3-7) cycloalkyl; Perhaps two R
162The nitrogen coupled by covalent linkage combines, formation 5,6 or the saturated heterocycle of 7-person.
13. according to the compound of claim 12, wherein R
2Be selected from: aryl or Het, each group choose wantonly and are selected from following replacement of substituting group list or two replacements: halogen, haloalkyl, N
3, perhaps
A) (C
1-6) alkyl, it is optional by OH, O (C
1-6) alkyl or SO
2(C
1-6Alkyl) replaces;
B) (C
1-6) alkoxyl group;
E) NR
111R
112, R wherein
111And R
112Be H independently, (C
1-6) alkyl, (C
3-7) cycloalkyl, perhaps R
112Be 6-or 10-person's aryl, Het, (C
1-6) alkyl-aryl or (C
1-6) alkyl-Het; Perhaps two R
111And R
112The nitrogen coupled by covalent linkage combines, and forms nitrogenous heterocycle, described alkyl, and cycloalkyl, aryl, Het, each among alkyl-aryl or the alkyl-Het is optional to be replaced by following groups: halogen, perhaps
-OR
161Or N (R
162)
2, R wherein
161And each R
162Be H independently, (C
1-6) alkyl, perhaps two R
162The nitrogen coupled by covalent linkage combines, and forms nitrogenous heterocycle;
F) NHCOR
117, R wherein
117Be (C
1-6) alkyl, O (C
1-6) alkyl or O (C
3-7) cycloalkyl;
I) CO-aryl; And
K) CONH
2, CONH (C
1-6Alkyl), CON (C
1-6Alkyl)
2, CONH-aryl, or CONHC
1-6Alkyl-aryl.
14. according to the compound of claim 13, wherein R
2Be aryl or Het, each group is chosen wantonly and is selected from following replacement of substituting group list or two replacements: halogen, and haloalkyl, perhaps
A) (C
1-6) alkyl, it is optional by OH, O (C
1-6) alkyl or SO
2(C
1-6Alkyl) replaces;
B) (C
1-6) alkoxyl group; And
E) NR
111R
112, R wherein
111And R
112Be H independently, (C
1-6) alkyl, (C
3-7) cycloalkyl, perhaps R
112Be 6-or 10-person's aryl, Het, (C
1-6) alkyl-aryl or (C
1-6) alkyl-Het; Perhaps two R
111And R
112The nitrogen coupled by covalent linkage combines, and forms nitrogenous heterocycle, described alkyl, and cycloalkyl, aryl, Het, each among alkyl-aryl or the alkyl-Het is all randomly replaced by following groups: halogen, perhaps
-OR
161Or N (R
162)
2, R wherein
161And each R
162Be H independently, (C
1-6) alkyl, perhaps two R
162The nitrogen coupled by covalent linkage combines, and forms nitrogenous heterocycle.
15. according to the compound of claim 14, wherein R
2For phenyl or be selected from following heterocycle:
16. according to the compound of claim 15, wherein R
2Be selected from:
H，Br，CH＝CH
2，
And
17. according to the compound of claim 16, wherein R
2Be selected from:
18. according to the compound of claim 17, wherein R
2Be selected from:
19. according to the compound of claim 1, wherein R
3Be selected from: (C
3-7) cycloalkyl, (C
3-7) cycloalkenyl group, (C
6-10) bicyclic alkyl, (C
6-10) bicycloenyl, 6-or 10-person's aryl, or Het.
20. according to the compound of claim 19, wherein R
3Be (C
3-7) cycloalkyl.
21. according to the compound of claim 20, wherein R
3Be cyclopentyl, or cyclohexyl.
22. according to the compound of claim 1, wherein Y
1Be O.
23. according to the compound of claim 1, wherein Z is OR
6, R wherein
6Be H, (C
1-6) alkyl, it is chosen wantonly and is replaced by following groups: halogen, hydroxyl, carboxyl, amino, C
1-6Alkoxyl group, C
1-6Alkoxy carbonyl, and C
1-6Alkylamino; Perhaps R
6Be C
1-6Alkylaryl, it is chosen wantonly and is replaced by following groups: halogen, cyano group, nitro, C
1-6Alkyl, C
1-6Haloalkyl, C
1-6Alkyloyl ,-(CH
2)
1-6-COOR
7,-(CH
2)
1-6-CONR
7R
8,-(CH
2)
1-6-NR
7R
8,-(CH
2)
1-6-NR
7COR
8,-(CH
2)
1-6-NHSO
2R
7,-(CH
2)
1-6-OR
7,-(CH
2)
1-6-SR
7,-(CH
2)
1-6-SO
2R
7, and-(CH
2)
1-6-SO
2NR
7R
8, each R wherein
7And each R
8Be H or C
1-6Alkyl,
Perhaps Z is NR
9R
10, each R wherein
9And R
10Be selected from: H, C
1-6Alkoxyl group, or C
1-6Alkyl, it is optional by halogen, hydroxyl, carboxyl, amino, C
1-6Alkoxyl group, C
1-6Alkoxy carbonyl, and C
1-6Alkylamino replaces.
24. according to the compound of claim 23, wherein Z is OH or O (C
1-6Alkyl), perhaps Z is NR
9R
10, R wherein
9Be H, and R
10Be H or C
1-6Alkyl.
25. according to the compound of claim 24, wherein Z is OH.
26. be selected from the compound of following formula:
A in the formula, R
2, R
3And Z is defined as follows:
27. be selected from the compound of following formula:
A in the formula, R
2, R
3Be defined as follows with Z:
28. the compound shown in the following formula Ia:
In the formula:
A is O, S, NR
1, or CR
1
B is NR
3Or CR
3
R
1Be selected from: H, (C
1-6) alkyl, benzyl, (C
1-6Alkyl)-(C
6-10Aryl), (C
1-6Alkyl)-5-or 6-person have 1～4 heteroatomic heterocycle that is selected from O, N and S, and have 1～4 heteroatomic 5-or 6-element heterocycle that is selected from O, N and S,
Wherein said benzyl and described heteroatoms are optional to be selected from COOH, COO (C by 1～4
1-6Alkyl), halogen, and (C
1-6Alkyl) substituting group replaces;
R
2Be selected from: H, halogen, (C
1-6) alkyl, (C
3-7) cycloalkyl, phenyl has 1～4 heteroatomic 5-or 6-element heterocycle that is selected from O, N and S, pyridine-N-oxide, and have 1～4 heteroatomic 9-or assorted two ring of 10-person that are selected from O, N and S,
Described phenyl, heterocycle and assorted two rings are chosen wantonly and are selected from halogen, C (halogen) by 1～4
3, (C
1-6) alkyl, OH, O (C
1-6Alkyl), NH
2, and N (C
1-6Alkyl)
2Substituting group replace;
R
3Be selected from: have 1～4 heteroatomic 5-that is selected from O, N and S, 6-or 7-element heterocycle, norcamphane, (C
3-7) cycloalkyl and (C
3-7) cycloalkyl-(C
1-6Alkyl);
M is N, CR
4, or COR
5, R wherein
4Be selected from: H, halogen, and (C
1-6Alkyl); And R
5Be selected from: H and (C
1-6Alkyl);
K and L are N or CH;
-----represent singly-bound or two key;
Y is O;
Z is OR
6Or NR
6R
6a, R wherein
6Be selected from: H, (C
1-6) alkyl, wherein said alkyl is optional to be selected from OH, COOH, COO (C by 1～4
1-6) alkyl, (C
1-6) substituting group of alkyl replaces, alkyl described later is optional to be selected from COOH, NHCO (C by 1～4
1-6Alkyl), NH
2, NH (C
1-6Alkyl), reach N (C
1-6Alkyl)
2Substituting group replace;
Or its salt.
29. the inhibitor that duplicates as HCV according to the compound or pharmaceutically acceptable salt thereof of the formula I of claim 1.
30. the pharmaceutical composition that treatment or prevention HCV infect comprises the compound or pharmaceutically acceptable salt thereof according to the formula I of claim 1 of significant quantity and pharmaceutically acceptable carrier.
31., also comprise immunomodulator according to the pharmaceutical composition of claim 30.
32. according to the pharmaceutical composition of claim 31, wherein said immunomodulator is selected from: α-, β-, δ-γ-, and ω-Interferon, rabbit.
33., also comprise virazole or amantadine according to the pharmaceutical composition of claim 30.
34., also comprise other HCV AG14361 according to the pharmaceutical composition of claim 30.
35., also comprise and be selected from helicase, polysaccharase, the inhibitor of other HCV target of metalloprotease and IRES according to the pharmaceutical composition of claim 34.
36. following formula (1a) or intermediate (1b):
A in the formula, B, K, L, and M is as defined in claim 1, and PG is H or carboxyl-protecting group, X is a metal.
37. a method for preparing following formula compound (iii),
A in the formula, R
2, B, K, L, M, PG and X such as claim 36 are defined, comprising:
A), under the existence of alkali and additive, in suitable solvent, make the intermediate (1a) and the R of claim 36 at metal catalyst
2-X coupling.
38. a method for preparing following formula compound (iii),
A in the formula, R
2, B, K, L, M, and PG such as claim 36 definition, comprising:
A), under the existence of alkali and additive, in suitable solvent, make the intermediate (1b) and the R of claim 36 at metal catalyst
2-X ' coupling, wherein X ' such as claim 36 definition.
39. according to the method for claim 37 or 38, wherein said metal catalyst is selected from: Pd, Ni, Ru and Cu.
40. according to the method for claim 37 or 38, wherein said additive is selected from: phosphine ligand, Cu salt, Li salt, ammonium salt and CsF.
41. according to the method for claim 37 or 38, wherein said metal is selected from: Li, Sn (C
1-6Alkyl)
3, Sn (aryl)
3, B (OH)
2, B (OC
1-6Alkyl)
2And metal halide.
42. the application of compound in the medicine that preparation treatment HCV infects according to the formula I of claim 1.
Applications Claiming Priority (3)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| US30767401P | 2001-07-25 | 2001-07-25 | |
| US60/307,674 | 2001-07-25 | ||
| US60/338,061 | 2001-12-07 |
Related Parent Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CNB028187970A Division CN100443469C (en) | 2001-07-25 | 2002-07-18 | Viral polymerase inhibitors |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN101302182A true CN101302182A (en) | 2008-11-12 |
Family
ID=34572608
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CNA2008101085783A Pending CN101302182A (en) | 2001-07-25 | 2002-07-18 | Virus polymerase inhibitors |
Country Status (4)
| Country | Link |
|---|---|
| CN (1) | CN101302182A (en) |
| EC (1) | ECSP044958A (en) |
| UA (2) | UA101465C2 (en) |
| ZA (2) | ZA200400122B (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102427726A (en) * | 2009-03-27 | 2012-04-25 | 普雷西迪奥制药公司 | Fused ring inhibitors of hepatitis c |
-
2002
- 2002-07-18 CN CNA2008101085783A patent/CN101302182A/en active Pending
- 2002-07-18 UA UAA200809972A patent/UA101465C2/en unknown
- 2002-07-18 UA UA2004021314A patent/UA85366C2/en unknown
-
2004
- 2004-01-08 ZA ZA200400122A patent/ZA200400122B/en unknown
- 2004-01-13 ZA ZA200400240A patent/ZA200400240B/en unknown
- 2004-01-23 EC EC2004004958A patent/ECSP044958A/en unknown
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102427726A (en) * | 2009-03-27 | 2012-04-25 | 普雷西迪奥制药公司 | Fused ring inhibitors of hepatitis c |
Also Published As
| Publication number | Publication date |
|---|---|
| ECSP044958A (en) | 2004-03-23 |
| ZA200400240B (en) | 2004-11-01 |
| ZA200400122B (en) | 2004-11-03 |
| UA85366C2 (en) | 2009-01-26 |
| UA101465C2 (en) | 2013-04-10 |
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| CN100443469C (en) | Viral polymerase inhibitors | |
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| AU2002355150A1 (en) | Hepatitis C virus polymerase inhibitors with heterobicyclic structure | |
| CN101302182A (en) | Virus polymerase inhibitors |
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