CN103930109B

CN103930109B - The substituted heterocyclyl amides of N-

Info

Publication number: CN103930109B
Application number: CN201280055623.8A
Authority: CN
Inventors: K·J·巴拉; E·巴德; L·爱德华兹; C·豪沙姆; D·M·莱格兰德; R·J·泰勒
Original assignee: Novartis AG
Current assignee: Novartis AG
Priority date: 2011-09-16
Filing date: 2012-09-14
Publication date: 2016-11-30
Anticipated expiration: 2032-09-14

Abstract

The present invention relates to compound of formula I and officinal salt thereof and solvate, wherein R¹、R²、R³、R^a, A, B, D and E as herein defined.The activity of compound regulation CFTR, and can be used for treating inflammatory or obstructive airway diseases or mucosal hydration, including such as cystic fibrosis.Also describe the pharmaceutical composition of inclusion compound and prepare the method for compound.

Description

The substituted heterocyclyl amides of N-

The present invention relates to the substituted heterocyclyl amides of N-, their preparation and the purposes as medicine.Specifically, N-takes The heterocyclyl amides regulation CFTR in generation is active and can be used for treating inflammatory or obstructive airway diseases or mucosal hydration, including example Such as cystic fibrosis.

On the one hand, the invention provides compound of formula I:

Or its officinal salt or solvate, wherein:

A is N or CR⁴；

B is N or CR⁵；

D is N or CR⁶；

E is N or CR⁷, condition be D and E be not all N；

R¹Selected from H；The C optionally replaced by one or more halogen atoms₁-C₈Alkyl；C₂-C₈Alkenyl；C₂-C₈Alkynyl； C₃-C₁₀Cycloalkyl；C₅-C₁₀Cycloalkenyl group；-C₁-C₄Alkyl-C₃-C₈Cycloalkyl；Optionally replaced by one or more halogen atoms C₁-C₈Alkoxyl；OH；Halogen；SO₂NR⁸R⁹；SO₂R¹⁰；The S-C optionally replaced by one or more halogen atoms₁-C₈Alkyl；Appoint Select the S-C replaced by one or more Z substituent groups₆-C₁₄Aryl；-(C₀-C₄Alkyl)-C₆-C₁₄Aryl；-(C₀-C₄Alkyl)-3 to 14 yuan of heterocyclic radicals, wherein heterocyclic radical comprises at least one hetero atom selected from N, O and S；CN；NR¹¹R¹²；CONR¹³R¹⁴； NR¹³SO2R¹⁵；NR¹³C(O)R¹⁵And CO₂R¹⁵, wherein cycloalkyl, cycloalkenyl group, aryl and heterocyclic radical are the most one or more Z substituent group replaces；

R²Selected from the C optionally replaced by one or more halogen atoms₁-C₈Alkyl；C₂-C₈Alkenyl； C₂-C₈Alkynyl；C₃- C₁₀Cycloalkyl；C₅-C₁₀Cycloalkenyl group；-C₁-C₄Alkyl-C₃-C₈Cycloalkyl；The C optionally replaced by one or more halogen atoms₁-C₈ Alkoxyl；OH；Cl；Br；I；SO₂NR⁸R⁹；SO₂R¹⁰；The S-C optionally replaced by one or more halogen atoms₁-C₈Alkyl；S- C₆-C₁₄Aryl；-(C₀-C₄Alkyl)-C₆-C₁₄Aryl；-(C₀-C₄Alkyl)-3 to 14 yuan of heterocyclic radicals, wherein heterocyclic radical comprises at least One hetero atom selected from N, O and S；CN；CONR¹³R¹⁴；NR¹³SO2R¹⁵；NR¹³C(O)R¹⁵And CO₂R¹⁵, wherein cycloalkyl, cyclenes Base, aryl and heterocyclic radical are the most optionally replaced by one or more Z substituent groups；

R^aSelected from H；The C optionally replaced by one or more halogen atoms₁-C₈Alkyl；C₂-C₈Alkenyl；C₂-C₈Alkynyl； C₃-C₁₀Cycloalkyl；C₅-C₁₀Cycloalkenyl group；-C₁-C₄Alkyl-C₃-C₈Cycloalkyl；SO₂R¹⁰；-(C₀-C₄Alkyl)-C₆-C₁₄Aryl；- (C₀-C₄Alkyl)-3 to 14 yuan of heterocyclic radicals, wherein heterocyclic radical comprises at least one hetero atom selected from N, O and S；C(O)R¹⁵With CO₂R¹⁵, wherein cycloalkyl, cycloalkenyl group, aryl and heterocyclic radical are the most optionally replaced by one or more Z substituent groups；

R³And R⁴It is each independently selected from H and the C optionally replaced by one or more halogen atoms₁-C₈Alkyl；

R⁵Selected from H；The C optionally replaced by one or more halogen atoms₁-C₈Alkyl；C₂-C₈Alkenyl；C₂-C₈Alkynyl； C₃-C₁₀Cycloalkyl；C₅-C₁₀Cycloalkenyl group；-C₁-C₄Alkyl-C₃-C₈Cycloalkyl；Halogen；-(C₀-C₄Alkyl)-C₆-C₁₄Aryl；With- (C₀-C₄Alkyl)-3 to 14 yuan of heterocyclic radicals, wherein heterocyclic radical comprises at least one hetero atom selected from N, O and S；Wherein cycloalkyl, Cycloalkenyl group, aryl and heterocyclic radical are the most optionally replaced by one or more Z substituent groups；

R⁶And R⁷It is each independently selected from H；The C optionally replaced by one or more halogen atoms₁-C₈Alkyl；C₂-C₈Alkene Base；C₂-C₈Alkynyl；C₃-C₁₀Cycloalkyl；C₅-C₁₀Cycloalkenyl group；-C₁-C₄Alkyl-C₃-C₈Cycloalkyl；Optionally by one or more halogen The element substituted C of atom₁-C₈Alkoxyl；OH；CN；Halogen；-(C₀-C₄Alkyl)-C₆-C₁₄Aryl；-(C₀-C₄Alkyl)-3 to 14 yuan Heterocyclic radical, wherein heterocyclic radical comprises at least one hetero atom selected from N, O and S；With-(C₀-C₄Alkyl)-CO₂R¹⁵, wherein cycloalkanes Base, cycloalkenyl group, aryl and heterocyclic radical are the most optionally replaced by one or more Z substituent groups；Or

R⁶And R⁷It is following formula group independently of one another:

-(CH₂)_n-X-(CH₂)_m-NR¹⁷R¹⁸；Or

R⁶And R⁷Carbon atom in connection formed together 5 to 8 yuan of carbocyclic ring systems or containing one or more selected from N, O and Heteroatomic 5 to 8 yuan of heterocycle systems of S, wherein ring system is optionally replaced by one or more Z substituent groups；

X be do not exist, O, S, CO, SO, SO₂Or CH₂；

N and m is each independently selected from 0,1,2 and 3；

R⁸、R¹¹、R¹³And R¹⁷It is each independently selected from H, the C optionally replaced by one or more halogen atoms₁-C₈Alkyl, C₃-C₁₀Cycloalkyl and-(C₁-C₄Alkyl)-C₃-C₈Cycloalkyl；

R⁹、R¹⁰、R¹²、R¹⁴、R¹⁵、R¹⁶And R¹⁸It is each independently selected from H；Optionally replaced by one or more halogen atoms C₁-C₈Alkyl；C₂-C₈Alkenyl；C₂-C₈Alkynyl；C₃-C₁₀Cycloalkyl；C₅-C₁₀Cycloalkenyl group；-C₁-C₄Alkyl-C₃-C₈Cycloalkyl；- (C₀-C₄Alkyl)-C₆-C₁₄Aryl；With-(C₀-C₄Alkyl)-3 to 14 yuan of heterocyclic radicals, wherein heterocyclic radical comprises at least one and is selected from The hetero atom of N, O and S, wherein cycloalkyl, cycloalkenyl group, aryl and heterocyclic radical are the most optionally replaced by one or more Z substituent groups； Or

R⁸And R⁹、R¹¹And R¹²、R¹³And R¹⁴And R¹⁷And R¹⁸Nitrogen-atoms in connection can be formed together optionally by one Individual or multiple substituted 4 to 14 yuan of heterocyclic radicals of Z substituent group；

Z is independently selected from OH；Aryl；O-aryl；Benzyl；O-benzyl；Optionally by one or more OH groups or NH₂Group Substituted C₁-C₆Alkyl；The C optionally replaced by one or more halogen atoms₁-C₆Alkyl；Optionally by one or more OH groups Or C₁-C₄The substituted C of alkoxyl₁-C₆Alkoxyl；NR¹⁸(SO₂)R²¹；(SO₂)NR¹⁹R²¹；(SO₂)R²¹；NR¹⁸C(O)R²¹；C(O) NR¹⁹R²¹；NR¹⁹C(O)NR¹⁹R²¹；NR¹⁸C(O)OR¹⁹；NR¹⁹R²¹；C(O)OR¹⁹；C(O)R¹⁹；SR¹⁹；OR¹⁹；Oxo；CN；NO₂；Halogen Element；With-(C₀-C₄Alkyl)-3 to 14 yuan of heterocyclic radicals, wherein heterocyclic radical comprises at least one hetero atom selected from N, O and S, optionally By one or more selected from halogen, oxo, C₁-C₆Alkyl and C (O) C₁-C₆The group of alkyl replaces；

R¹⁸And R²⁰It is each independently selected from H and C₁-C₆Alkyl；

R¹⁹And R²¹It is each independently selected from H；C₁-C₈Alkyl；C₃-C₈Cycloalkyl；C₁-C₄Alkoxy-C₁-C₄Alkyl；Optionally By one or more selected from C₁-C₆Alkyl, C₁-C₆Substituted (the C of group of alkoxyl and halogen₀-C₄Alkyl)-aryl；(C₀-C₄Alkane Base)-3-to 14-unit heterocyclic radical, heterocyclic radical comprises one or more hetero atom selected from N, O and S, optionally by one or more choosings From halogen, oxo, C₁-C₆Alkyl and C (O) C₁-C₆The group of alkyl replaces；Optionally by one or more selected from C₁-C₆Alkyl, C₁-C₆Substituted (the C of group of alkoxyl and halogen₀-C₄Alkyl)-O-aryl；(C₀-C₄Alkyl)-O-3-to 14-unit heterocycle Base, heterocyclic radical comprises one or more hetero atom selected from N, O and S, optionally by one or more selected from halogen, C₁-C₆Alkyl and C(O)C₁-C₆The group of alkyl replaces；Wherein alkyl is optionally by one or more halogen atoms, C₁-C₄Alkoxyl, C (O) NH₂、C (O)NHC₁-C₆Alkyl or C (O) N (C₁-C₆Alkyl)₂Replace；Or

R¹⁹And R²¹Nitrogen-atoms in connection forms 5-to 10-unit heterocyclic radical together, and heterocyclic radical comprises one or more Selected from the further hetero atom of N, O and S, heterocyclic radical is optionally replaced selected from following substituent group by one or more: OH；Halogen Element；Aryl；Comprise one or more heteroatomic 5-to 10-unit heterocyclic radical selected from N, O and S；S(O)₂-aryl；S(O)₂-C₁- C₆Alkyl；The C optionally replaced by one or more halogen atoms₁-C₆Alkyl；Optionally by one or more OH groups or C₁-C₄Alkane The substituted C of epoxide₁-C₆Alkoxyl；With C (O) OC₁-C₆Alkyl, wherein aryl and heterocyclic substituent self are optionally by C₁-C₆Alkyl, C₁-C₆Haloalkyl or C₁-C₆Alkoxyl replaces；

It is for treating disease or the obstacle of CFTR mediation.

Second aspect, the invention provides compound of formula I:

Or its officinal salt or solvate, wherein:

A is N or CR⁴；

B is N or CR⁵；

D is N or CR⁶；

E is N or CR⁷, condition is:

I) D and E is not all N；And

Ii) one or both of A or B is N；

R¹Selected from H；The C optionally replaced by one or more halogen atoms₁-C₈Alkyl；C₂-C₈Alkenyl；C₂-C₈Alkynyl； C₃-C₁₀Cycloalkyl；C₅-C₁₀Cycloalkenyl group；-C₁-C₄Alkyl-C₃-C₈Cycloalkyl；Optionally replaced by one or more halogen atoms C₁-C₈Alkoxyl；OH；Halogen； SO₂NR⁸R⁹；SO₂R¹⁰；The S-C optionally replaced by one or more halogen atoms₁-C₈Alkyl； The S-C optionally replaced by one or more Z substituent groups₆-C₁₄Aryl；-(C₀-C₄Alkyl)-C₆-C₁₄Aryl；-(C₀-C₄Alkyl)-3 To 14 yuan of heterocyclic radicals, wherein heterocyclic radical comprises at least one hetero atom selected from N, O and S；CN；NR¹¹R¹²；CONR¹³R¹⁴； NR¹³SO2R¹⁵；NR¹³C(O)R¹⁵And CO₂R¹⁵, wherein cycloalkyl, cycloalkenyl group, aryl and heterocyclic radical are the most one or more Z substituent group replaces；

R²Selected from the C optionally replaced by one or more halogen atoms₁-C₈Alkyl；C₂-C₈Alkenyl；C₂-C₈Alkynyl；C₃- C₁₀Cycloalkyl；C₅-C₁₀Cycloalkenyl group；-C₁-C₄Alkyl-C₃-C₈Cycloalkyl；The C optionally replaced by one or more halogen atoms₁-C₈ Alkoxyl；OH；Cl；Br；I；SO₂NR⁸R⁹；SO₂R¹⁰；The S-C optionally replaced by one or more halogen atoms₁-C₈Alkyl；S- C₆-C₁₄Aryl；-(C₀-C₄Alkyl)-C₆-C₁₄Aryl；-(C₀-C₄Alkyl)-3 to 14 yuan of heterocyclic radicals, wherein heterocyclic radical comprises at least One hetero atom selected from N, O and S；CN；CONR¹³R¹⁴；NR¹³SO2R¹⁵；NR¹³C(O)R¹⁵And CO₂R¹⁵, wherein cycloalkyl, cyclenes Base, aryl and heterocyclic radical are the most optionally replaced by one or more Z substituent groups；

R⁶And R⁷It is each independently selected from H；The C optionally replaced by one or more halogen atoms₁-C₈Alkyl；C₂-C₈Alkene Base；C₂-C₈Alkynyl；C₃-C₁₀Cycloalkyl；C₅-C₁₀Cycloalkenyl group；-C₁-C₄Alkyl-C₃-C₈Cycloalkyl；Optionally by one or more halogen The element substituted C of atom₁-C₈Alkoxyl；OH； CN；Halogen；-(C₀-C₄Alkyl)-C₆-C₁₄Aryl；-(C₀-C₄Alkyl)-3 to 14 yuan Heterocyclic radical, wherein heterocyclic radical comprises at least one hetero atom selected from N, O and S；With-(C₀-C₄Alkyl)-CO₂R¹⁵, wherein cycloalkanes Base, cycloalkenyl group, aryl and heterocyclic radical are the most optionally replaced by one or more Z substituent groups；Or

R⁶And R⁷It is following formula group independently of one another:

-(CH₂)_n-X-(CH₂)_m-NR¹⁷R¹⁸；Or

R⁶And R⁷Carbon atom in connection forms 5 to 8 yuan of carbocyclic ring systems or together containing one or more selected from N, O and S Heteroatomic 5 to 8 yuan of heterocycle systems, wherein ring system is optionally replaced by one or more Z substituent groups；

X be do not exist, O, S, CO, SO, SO₂Or CH₂；

N and m is each independently selected from 0,1,2 and 3；

R⁹、R¹⁰、R¹²、R¹⁴、R¹⁵、R¹⁶And R¹⁸It is each independently selected from H；Optionally replaced by one or more halogen atoms C₁-C₈Alkyl；C₂-C₈Alkenyl；C₂-C₈Alkynyl；C₃-C₁₀Cycloalkyl；C₅-C₁₀Cycloalkenyl group；-C₁-C₄Alkyl-C₃-C₈Cycloalkyl；- (C₀-C₄Alkyl)-C₆-C₁₄Aryl；With-(C₀-C₄Alkyl)-3 to 14 yuan of heterocyclic radicals, wherein heterocyclic radical comprises at least one and is selected from The hetero atom of N, O and S, wherein cycloalkyl, cycloalkenyl group, aryl and heterocyclic radical are the most optionally replaced by one or more Z substituent groups； Or R⁸And R⁹、R¹¹And R¹²、R¹³And R¹⁴And R¹⁷And R¹⁸Nitrogen-atoms in connection can be formed together optionally by one or many Substituted 4 to the 14 yuan of heterocyclic radicals of individual Z substituent group；

Z is independently selected from OH；Aryl；O-aryl；Benzyl；O-benzyl；Optionally by one or more OH groups or NH₂Group Substituted C₁-C₆Alkyl；The C optionally replaced by one or more halogen atoms₁-C₆Alkyl；Optionally by one or more OH groups Or C₁-C₄The substituted C of alkoxyl₁-C₆Alkoxyl；NR^18’(SO₂)R²¹；(SO₂)NR¹⁹R²¹；(SO₂)R²¹；NR^18’C(O)R²¹；C(O) NR¹⁹R²¹；NR¹⁹C(O)NR¹⁹R²¹；NR^18’C(O)OR¹⁹；NR¹⁹R²¹；C(O)OR¹⁹；C(O)R¹⁹；SR¹⁹；OR¹⁹；Oxo；CN；NO₂； Halogen；With-(C₀-C₄Alkyl)-3 to 14 yuan of heterocyclic radicals, wherein heterocyclic radical comprises at least one hetero atom selected from N, O and S, appoints Select by one or more selected from halogen, oxo, C₁-C₆Alkyl and C (O) C₁-C₆The group of alkyl replaces；

R^18’And R²⁰It is each independently selected from H and C₁-C₆Alkyl；

R¹⁹And R²¹It is each independently selected from H；C₁-C₈Alkyl；C₃-C₈Cycloalkyl；C₁-C₄Alkoxy-C₁-C₄Alkyl；Optionally By one or more selected from C₁-C₆Alkyl, C₁-C₆Substituted (the C of group of alkoxyl and halogen₀-C₄Alkyl)-aryl；(C₀-C₄Alkane Base)-3-to 14-unit heterocyclic radical, heterocyclic radical comprises one or more hetero atom selected from N, O and S, optionally by one or more choosings From halogen, oxo, C₁-C₆Alkyl and C (O) C₁-C₆The group of alkyl replaces；Optionally by one or more selected from C₁-C₆Alkyl, C₁-C₆Alkoxyl and the (C of halogen substiuted₀-C₄Alkyl)-O-aryl；(C₀-C₄Alkyl)-O-3-to 14-unit heterocyclic radical, heterocycle Base comprises one or more hetero atom selected from N, O and S, optionally by one or more selected from halogen, C₁-C₆Alkyl and C (O) C₁- C₆The group of alkyl replaces；Wherein alkyl is optionally by one or more halogen atoms, C₁-C₄Alkoxyl, C (O) NH₂、C(O)NHC₁- C₆Alkyl or C (O) N (C₁-C₆Alkyl)₂Replace；Or

R¹⁹And R²¹Forming 5-to 10-unit heterocyclic radical together with the nitrogen-atoms being connected with them, heterocyclic radical includes one or more Selected from the further hetero atom of N, O and S, heterocyclic radical is optionally replaced selected from following substituent group by one or more: OH；Halogen Element；Aryl；Comprise one or more selected from N, O and S heteroatomic 5-to 10-unit heterocyclic radical；S(O)₂-aryl；S(O)₂-C₁-C₆ Alkyl；The C optionally replaced by one or more halogen atoms₁-C₆Alkyl；Optionally by one or more OH groups or C₁-C₄Alcoxyl The substituted C of base₁-C₆Alkoxyl；With C (O) OC₁-C₆Alkyl, wherein aryl and heterocyclic substituent self are optionally by C₁-C₆Alkyl, C₁-C₆Haloalkyl or C₁-C₆Alkoxyl replaces.

Unless specifically stated otherwise, otherwise term " the compounds of this invention " refers to formula (I) compound and subformula thereof, its The salt of prodrug, compound and/or prodrug, the hydrate of compound, salt and/or prodrug or solvate, and stereoisomer (including diastereomer and enantiomer), tautomer and isotope-labeled compound (including that deuterium replaces), with And the part (such as polymorph, solvate and/or hydrate) that inherence is formed.

In order to the purpose of the application is described, apply following definition, and time applicable, the term that odd number uses also includes multiple Number, and vice versa.

" optionally substituted " represents that the group mentioned can be in one or more positions by any one base of listed thereafter The combination of group or any group replaces.

" optionally replaced by one or more Z group " and represent that relevant group can comprise one or more substituent group, often The group that individual substituent group includes in defining independently selected from Z.Therefore, when there is two or more Z group substituent group, they Can be same or different.

" halogen " used herein can be fluorine, chlorine, bromine or iodine.

" C used herein₁-C₈-alkyl " represent the straight or branched alkyl with 1-8 carbon atom.If carbon atom Varying number is specific, such as C₆Or C₃, then correspondingly revise this definition.

" C used herein₁-C₈-alkoxyl " represent the straight or branched alkoxyl with 1-8 carbon atom.If carbon is former The varying number of son is specific, such as C₆Or C₃, then correspondingly revise this definition.

Term " alkylidene " represents the straight or branched saturated hydrocarbon chain containing 1 to 8 carbon atom.If the difference of carbon atom Quantity is specific, such as C₆Or C₃, then correspondingly revise this definition.

" amino-C₁-C₈-alkyl " and " amino-C₁-C₈-alkoxyl " represent be connected to C by nitrogen-atoms₁-C₈-alkyl Amino, such as NH₂-(C₁-C₈)-, or be connected to C by nitrogen-atoms₁-C₈The amino of-alkoxyl, such as NH₂-(C₁-C₈)-O-.As Really the varying number of carbon atom is specific, such as C₆Or C₃, then correspondingly revise this definition.

“C₁-C₈-alkyl amino " or " two (C₁-C₈-alkyl) amino " represent and be connected to the fixed above of amino by carbon atom The C of justice₁-C₈-alkyl.Two (C₁-C₈-alkyl) C in amino₁-C₈-alkyl can be same or different.If carbon atom Varying number be specific, such as C₆Or C₃, then correspondingly revise this definition.

" amino-(hydroxyl)-C₁-C₈-alkyl " represent be connected to C by nitrogen-atoms₁-C₈The amino of-alkyl and former by oxygen Son is connected to identical C₁-C₈The hydroxyl of-alkyl.If the varying number of carbon atom is specific, such as C₆Or C₃, then phase This definition should be revised in ground.

" C used herein₁-C₈-alkyl-carbonyl " and " C₁-C₈-alkoxy carbonyl " represent be connected to by carbon atom respectively The C defined above of carbonyl₁-C₈-alkyl or C₁-C₈-alkoxyl.If the varying number of carbon atom is specific, such as C₆Or C₃, then correspondingly revise this definition.

" C used herein₃-C₈-naphthene base carbonyl " represent the C defined above being connected to carbonyl by carbon atom₃-C₈- Cycloalkyl.If the varying number of carbon atom is specific, such as C₆Or C₃, then correspondingly revise this definition.

" C used herein₇-C₁₄-aralkyl " represent by C defined herein₆-C₁₀On-aromatic carbocyclic group is substituted The alkyl of literary composition definition, such as C₁-C₄-alkyl.If the varying number of carbon atom is specific, such as C₆Or C₃, then correspondingly Revise this definition.

" C used herein₃-C₁₅-carbocylic radical " represent the carbocyclic ring with the saturated of 3-to 15-ring carbon atom or fractional saturation Group, such as C₃-C₈-cycloalkyl.C₃-C₁₅The example of-carbocylic radical includes but not limited to cyclopropyl, cyclobutyl, cyclopenta, hexamethylene Base, suberyl or ring octyl group, or two ring groups, such as bicyclooctyl, diazabicyclononyl include indanyl and indenyl and bicyclo-decyl. If the varying number of carbon atom is specific, such as C₆, then correspondingly revise this definition.

" aryl " used herein or " C₆-C₁₅-aromatic carbon ring base " represent the fragrance with 6-to 15-ring carbon atom Base.C₆-C₁₅The example of-aromatic carbon ring base include but not limited to phenyl, phenylene, benzene three base (benzenetriyl), naphthyl, Naphthylene, naphthalene three base (naphthalenetriyl) or anthrylene.If the varying number of carbon atom is specific, such as C₁₀, The most correspondingly revise this definition.

" 4-to 8-unit heterocyclic radical ", " 5-to 6-unit heterocyclic radical ", " 3-to 10-unit heterocyclic radical ", " 3-to 14-unit heterocycle Base ", " 4-to 14-unit heterocyclic radical " and " 5-to 14-unit heterocyclic radical " represent containing at least one respectively selected from nitrogen, oxygen and the ring of sulfur Heteroatomic 4-to 8-unit, 5-to 6-unit, 3-to 10-unit, 3-to 14-unit, 4-to 14-unit and 5-to 14-unit heterocycle, it is permissible It is saturated, fractional saturation or undersaturated (aromatic series).Heterocyclic radical includes monocycle base, condensed ring radical and bridging group.This kind of miscellaneous The example of ring group includes but not limited to furan, pyrroles, pyrrolidine, pyrazoles, imidazoles, triazole, different triazole, tetrazolium, thiadiazoles, different thiophene Azoles,Diazole, pyridine, piperidines, pyrazine,Azoles, differentAzoles, pyrazine, pyridazine, pyrimidine, piperazine, pyrrolidine, ketopyrrolidine, Morpholine, triazine,Piperazine, oxolane, Tetramethylene sulfide, tetrahydric thiapyran, Pentamethylene oxide., 1,4-bis-Alkane, 1,4-oxygen thia ring Hexane, indazole, quinoline, indazole, indole, 8-aza-bicyclo [3.2.1] octane or thiazole.

This document describes multiple embodiments of the present invention.Should be understood that the feature described in each embodiment can To combine with other specific features, to provide further embodiment.

In the embodiment of the present invention described Anywhere herein, A is N.

In the further embodiment of the present invention described Anywhere herein, R¹Selected from H；Optionally by one or many The substituted C of individual halogen atom₁-C₈Alkyl；Halogen；C₆-C₁₄Aryl；-(C₀-C₄Alkyl)-3 to 14 yuan of heterocyclic radical, wherein heterocyclic radicals Comprise at least one hetero atom selected from N, O and S；And NR¹¹R¹², wherein aryl and heterocyclic radical are the most optionally by one or more Z Substituent group replaces.

In the further embodiment of the present invention described Anywhere herein, R²It is optionally by one or more halogen The element substituted C of atom₁-C₈Alkyl；C₃-C₁₀Cycloalkyl；Or-C₁-C₄Alkyl-C₃-C₈Cycloalkyl.Compatibly, R²It is CF₃。

In the further embodiment of the present invention described Anywhere herein, R³It is H or methyl

In the further embodiment of the present invention described Anywhere herein, R^aIt is H.

In the further embodiment of the present invention described Anywhere herein, E is CR⁷。

Embodiment of the present invention defined above provide compound of formula I, wherein

A is N；

R¹Selected from H；Halogen；C₆-C₁₄Aryl；5 or 6-unit's heterocyclic radicals, wherein heterocyclic radical comprises at least one selected from N, O and S Hetero atom；And NR¹¹R¹², wherein aryl and heterocyclic radical are the most optionally replaced by one or more Z substituent groups.

R²It is CF₃；

R^aIt is H；

R³It is H or Me；And

E is CR⁷, other variablees the most all define the most Anywhere.

In another embodiment, the compounds of this invention is those that example below part is listed.

In another embodiment, the invention provides formula (I) compound, it is selected from:

3-amino-6-bromo-5-trifluoromethyl-pyrazine-2-formic acid (5-trifluoromethyl-2H-pyrazole-3-yl)-amide；

3-amino-6-bromo-5-trifluoromethyl-pyrazine-2-formic acid (5-cyclopropyl-2H-pyrazole-3-yl)-amide；

3-amino-6-bromo-5-trifluoromethyl-pyrazine-2-formic acid (the 5-tert-butyl group-2H-pyrazole-3-yl)-amide；

3-amino-6-bromo-5-trifluoromethyl-pyrazine-2-formic acid (5-methyl-2H-pyrazole-3-yl)-amide；

3-amino-6-bromo-5-trifluoromethylpyridin-2-formic acid (5-trifluoromethyl-2H-pyrazole-3-yl)-amide；

3-amino-6-bromo-5-trifluoromethyl-pyrazine-2-formic acid (2H-pyrazole-3-yl)-amide；

3-amino-6-bromo-5-trifluoromethylpyridin-2-formic acid (2H-pyrazole-3-yl)-amide；

3-amino-6-pyrrolidin-1-yl-5-trifluoromethyl-pyrazine-2-formic acid (5-trifluoromethyl-2H-pyrazole-3-yl)- Amide；

3-amino-6-morpholine-4-base-5-trifluoromethyl-pyrazine-2-formic acid (2H-pyrazole-3-yl)-amide；

3-amino-5-trifluoromethyl-pyrazine-2-formic acid (2H-pyrazole-3-yl)-amide；

5-[(3-amino-6-bromo-5-trifluoromethyl-pyrazine-2-carbonyl)-amino]-1H-[1,2,4] triazole-3-formic acid；

3-amino-6-dimethylamino-5-trifluoromethyl-pyrazine-2-formic acid (2H-pyrazole-3-yl)-amide；

3-amino-6-bromo-5-trifluoromethyl-pyrazine-2-formic acid (2H-[1,2,4] triazole-3-base) amide；

3-amino-6-chloro-5-trifluoromethyl-pyrazine-2-formic acid (2H-pyrazole-3-yl) amide；

3-amino-6-bromo-5-trifluoromethyl-pyrazine-2-formic acid (5-piperidin-1-yl methyl-2H-pyrazole-3-yl)-acyl Amine；

The bromo-N-of 3-amino-6-(1H-pyrazolo [4,3-b] pyrazine-3-base)-5-(trifluoromethyl) pyrazine-2-amide；

3-(3-amino-6-bromo-5-(trifluoromethyl) pyrazine-2-amide groups)-4,6-pyrrolin also [3,4-c] pyrazoles-5 (2H)-t-butyl formate；

The bromo-N-of 3-amino-6-(1H-pyrazolo [4,3-b] pyrazine-3-base)-5-(trifluoromethyl) picolinamide；

The bromo-N-of 3-amino-6-(4-(pyridine-2-base)-1H-pyrazoles-5-base)-5-(trifluoromethyl) pyrazine-2-amide；

The iodo-N-of 2-amino-5-(1H-pyrazoles-5-base)-4-(trifluoromethyl) benzamide hydrochloride salt；

The bromo-N-of 3-amino-6-(1H-pyrazolo [3,4-b] pyridin-3-yl)-5-(trifluoromethyl) pyrazine-2-amide；

The chloro-N-of 3-amino-6-(1H-pyrazoles-5-base)-5-(trifluoromethyl) picolinamide；

5-(3-amino-6-bromo-5-(trifluoromethyl) pyrazine-2-amide groups)-1H-1,2,4-triazole-3-methyl formate；

3-amino-N-(3-benzyl-1H-1,2,4-triazole-5-base)-6-bromo-5-(trifluoromethyl) pyrazine-2-amide；

3-amino-6-(4-fluorophenyl)-N-(1H-pyrazole-3-yl)-5-(trifluoromethyl) picolinamide；

3-amino-6-(4-chloro-2-methyl-phenyl)-5-trifluoromethylpyridin-2-formic acid (2H-pyrazole-3-yl)-acyl Amine；

5-amino-2 '-methoxyl group-3-trifluoromethyl-[2,3 '] bipyridyl-6-formic acid (1H-pyrazole-3-yl)-amide；

3-amino-6-[3-(2-oxo-pyrroli-1-ylmethyl)-phenyl]-5-trifluoromethylpyridin-2-formic acid (2H- Pyrazole-3-yl)-amide；

5-amino-6 '-methoxyl group-3-trifluoromethyl-[2,3 '] bipyridyl-6-formic acid (2H-pyrazole-3-yl)-amide；

3-amino-6-[4-(5-methyl-[1,3,4]Diazole-2-base)-phenyl]-5-trifluoromethylpyridin-2-first Acid (2H-pyrazole-3-yl)-amide；

3-amino-5,6-pair-trifluoromethyl-pyrazine-2-formic acid (4-methyl-2H-pyrazole-3-yl)-amide；

Or its officinal salt or solvate.

It should be appreciated that any for the present invention can be combined with other embodiment any with all of embodiment Thus the other embodiments of the present invention are described.Additionally, by any key element in embodiment and from any embodiment Other key element any and all of combine other embodiments are described.Skilled person would appreciate that can not The substituent group combination of energy the aspect of non-invention.

Unless the context otherwise requires, otherwise in this specification and subsequent claim, term " comprises " or it Version such as " includes " group being understood to refer to include described integer or step or integer or step, but does not arranges Except any other integer or step or integer or the group of step.

Term used herein " isomer " refers to have same molecular formula but atomic arrangement different chemical combination different with configuration Thing.Additionally, the given compound that term used herein " optical isomer " or " stereoisomer " refer to the present invention can exist Multiple stereoisomeric configurations in any one and include geometric isomer.It should be appreciated that substituent group can connect On the chiral centre of carbon atom.Term " chirality " represents have the molecule of not plyability on their mirror image companion, and art Language " achirality " expression is molecule that can be overlapping on their mirror image companion.Therefore, the present invention includes that the mapping of compound is different Structure body, diastereomer or racemic modification." enantiomer " is a pair stereoisomer that can not be overlapped mirror image each other.One 1:1 mixture to enantiomer is " raceme " mixture.Under appropriate circumstances, this term is used for referring to that raceme mixes Thing." diastereomer " but be the stereoisomer with at least two asymmetric atom mirror image the most each other.Absolute stereo Chemistry specifies according to Cahn-lngold-Prelog R-S system.When compound is pure enantiomer, each hands Spatial chemistry on property carbon can illustrate with R or S.The compound that its absolute configuration split is not clear can be according to them at sodium Rotate the direction (dextrorotation-or left-handed-) of linearly polarized light under D line wavelength and be designated as (+) or (-).As herein described some Compound contains one or more asymmetric center or axle, therefore can produce enantiomer, diastereomer and permissible Absolute stereochemical is defined as (R)-or (S)-other stereoisomeric forms in any ratio.

Depending on raw material and the method selected, compound can depend on that asymmetric carbon atom quantity is with a kind of possible Isomer or its form of mixtures exist, the purest optical isomer or isomer mixture, such as racemic modification and non-right Reflect isomer mixture.It is intended that include all these possible isomer, including racemic mixture, diastereo-isomerism Body mixture and the pure form of optically-active.(R) of optically active-and (S)-isomer can be prepared with chiral synthon or chiral reagent Or split with routine techniques.If compound contains double bond, then substituent group can be E or Z configuration.If compound contains Have dibasic cycloalkyl, then naphthenic substituent can have cis-or trans-configuration.Also include all tautomerism shapes Formula.

Term used herein " salt " represents acid-addition salts or the base addition salts of the compounds of this invention." salt " particularly including " officinal salt ".Term " officinal salt " represents biological effectiveness and the salt of character retaining the compounds of this invention, and it is usual It it not biology or other is undesirable.Under many circumstances, the compounds of this invention can by exist amino and/or carboxyl or Similar group forms acid and/or alkali salt.

Pharmaceutically acceptable acid addition salts can be formed with mineral acid and organic acid, such as acetate, aspartate, benzoic acid Salt, benzene sulfonate, bromide/hydrobromate, bicarbonate/carbonate, disulfate/sulfate, camsilate, chlorination Thing/hydrochlorate, chlortheophyllonate, citrate, ethanedisulphonate, fumarate, gluceptate, gluconic acid Salt, glucuronate, hippurate, hydriodate/iodide, isethionate, lactate, Lactobionate, dodecyl Sulfate, malate, maleate, malonate, mandelate, mesylate, Methylsulfate, naphthoate, naphthalene sulphur Hydrochlorate, nicotinate, nitrate, octadecanoate, oleate, oxalates, palmitate, embonate, phosphate/phosphor acid hydrogen Salt/dihydric phosphate, Polygalacturonate, propionate, stearate, succinate, sulfosalicylate, tartrate, Toluene fulfonate and trifluoroacetate.

Such as hydrochloric acid, hydrobromic acid, sulphuric acid, nitric acid, phosphoric acid etc. can be included from its derivative mineral acid obtaining salt.

Can from its derivative organic acid obtaining salt include such as acetic acid, propanoic acid, glycolic, oxalic acid, maleic acid, the third two Acid, succinic acid, fumaric acid, tartaric acid, citric acid, benzoic acid, mandelic acid, methanesulfonic acid, ethyl sulfonic acid, toluenesulfonic acid, sulfosalisylic Acid etc..Pharmaceutically acceptable base addition salts can be formed with inorganic and organic base.

Such as ammonium salt and the metal from periodic table of elements I-XII race can be included from its derivative inorganic base obtaining salt. In certain embodiments, salt is derived from sodium, potassium, ammonium, calcium, magnesium, ferrum, silver, zinc and copper；Particularly suitable salt include ammonium, potassium, Sodium, calcium and magnesium salt.

Such as primary, secondary and tertiary amine, substituted amine can be included, including naturally occurring from its derivative organic base obtaining salt Substituted amine, cyclic amine, deacidite etc..Some organic amine include isopropylamine, benzyl star salt (benzathine), Choline hydrochlorate (cholinate), diethanolamine, diethylamine, lysine, meglumine, piperazine and trometamol.

The officinal salt of the present invention can be synthesized by alkalescence or acidic moiety by conventional chemical method.Generally, permissible By making the free acid form of these compounds and stoichiometric applicable alkali (hydroxide of such as Na, Ca, Mg or K, carbon Hydrochlorate, bicarbonate etc.) react or by making the free alkali form of these compounds come with stoichiometric applicable acid reaction Prepare this kind of salt.This kind of reaction generally or in organic solvent or is carried out in water in both mixture.Generally, if cut Real feasible, then the use of non-aqueous media such as ether, ethyl acetate, ethanol, isopropanol or acetonitrile is desirable.Additionally The list of the salt being suitable for can be at such as " Remington ' s Pharmaceutical Sciences ", the 20th edition, Mack Publishing Company,Easton,Pa.,(1985)；And " the Handbook of of Stahl and Wermuth Pharmaceutical Salts:Properties, Selection and Use " (Wiley-VCH, Weinheim, Germany, 2002) find in.

Any structural formula given herein also aims to represent the unlabelled form of compound and isotope-labeled shape Formula.Isotope-labeled compound has the structure that structural formula given herein is described, the most one or more atoms Replaced by the atom with selected atomic mass or mass number.The isotopic example bag of the compounds of this invention can be mixed Include: the isotope of hydrogen, carbon, nitrogen, oxygen, phosphorus, fluorine and chlorine, be the most respectively²H、³H、¹¹C、¹³C、¹⁴C、¹⁵N、¹⁸F、³¹P、³²P、³⁵S、³⁶Cl、¹²⁵I.The present invention includes multiple isotope-labeled compound defined herein, such as, exist and mix radiosiotope (such as³H and¹⁴C) those, or mix non radioactive isotope (such as²H and¹³C) those.This kind of isotope-labeledization Compound can be used for metabolism research and (uses¹⁴C), kinetics research (such as uses²H or³H), detect or imaging technique, such as Positron emission tomography art (PET) or single photon emission computed tomography Imaging (SPECT), including medicine or substrate group Knit distributional analysis, or the radiation treatment for patient.Especially, PET or SPECT is studied,¹⁸F or the compound of labelling It is probably particularly desirable.Generally, can be by routine techniques well known by persons skilled in the art or by following described Embodiment and preparation example described in the isotope-labeled reagent that is suitable for of similar approach application replace the nonstandard of previously application Isotope-labeled formula (I) compound prepared by the reagent of note.

Additionally, by heavier isotope, particularly deuterium (i.e.²H or D) replace and can provide certain because of bigger metabolic stability Treat advantage, the Half-life in vivo such as increased or the volume requirements of reduction or the improvement of therapeutic index.It should be appreciated that Deuterium is considered the substituent group of formula (I) compound in the context of this article.The concentration of this kind of heavier isotope, particularly deuterium can To be defined as the isotope enrichment factor.Term used herein " the isotope enrichment factor " mean isotope abundance with specifically to The ratio of fixed natural abundance of isotopes.If the substituent group on the compounds of this invention is shown deuterium, the most this compound pair Have at least 3500 for each D-atom specified (on each D-atom specified, 52.5% deuterium mixes), at least 4000 (60% deuterium mix), at least 4500 (67.5% deuterium mixes), at least 5000 (75% deuterium mixes), at least 5500 (82.5% deuterium is mixed Enter), at least 6000 (90% deuterium mix), at least 6333.3 (95% deuterium mixes), at least 6466.7 (97% deuterium mixes), at least 6600 (99% deuterium mixes) or the isotope enrichment factor of at least 6633.3 (99.5% deuterium mixes).

The pharmaceutical acceptable solvates of the present invention includes that wherein recrystallisation solvent can be that isotope is substituted, such as D₂O、d₆-the third Ketone, d₆Those solvates of-DMSO.

The compounds of this invention, i.e. contain formula (I) the compound energy of the group that can work as hydrogen bond donor and/or receptor Eutectic is formed together with applicable eutectic (co-crystal) forming agent.Operation can be formed by formula (I) by known eutectic Compound prepares these eutectics.This generic operation includes grinding, heats, distillation, congruent melting or make formula (I) chemical combination under crystallization condition altogether The eutectic that thing contacts eutectic forming agent in the solution and separation is consequently formed.The eutectic forming agent being suitable for includes WO2004/ Those described in 078163.Therefore, present invention also offers the eutectic comprising formula (I) compound.

Any asymmetric carbon atom (such as carbon etc.) on the compounds of this invention can be with racemic modification or enantiomer The form of enrichment, such as (R)-, (S)-or (R, S)-configuration exist.In certain embodiments, each asymmetric atom exists (R)-or (S)-configuration in there is at least 50% enantiomer excess, at least 60% enantiomer excess, at least 70% right Reflect isomer excess, at least 80% enantiomer excess, at least 90% enantiomer excess, at least 95% enantiomer Excess or at least 99% enantiomer excess.If it is possible, the substituent group on atom with unsaturated double-bond can be with suitable Formula-(Z)-or trans-(E)-form exists.

Therefore, the compounds of this invention used herein can be possible isomer, rotamer, atropisomer, The form of one of tautomer or its mixture, such as, for the purest geometry (cis or trans) isomer, non-right Reflect isomer, optical isomer (enantiomer), racemic modification or its mixture.

The isomer mixture of any generation can be separated into pure or the purest according to the physics and chemistry difference of component Geometric isomer or optical isomer, diastereomer, racemic modification, such as, divided by chromatography and/or fractional crystallization From.

The end-product of any generation or the racemic modification of intermediate can be split into optical antimer by known method, Its diastereomeric salt such as obtained by the acid of separation optically active or alkali and the acidity or the alkali that discharge optically active Property compound splits.Especially, therefore basic moiety can be used for splitting into the compounds of this invention its optical antimer, Such as by the acid such as tartaric acid, dibenzoyl tartaric acid of fractional crystallization optically active, acetyl tartaric acid, two- O, O ' salt that formed of-p-toluoyl tartaric acid, mandelic acid, malic acid or Camphora-10-sulfonic acid splits.Racemic product is also Chiral chromatography can be passed through, such as use the high pressure lipuid chromatography (HPLC) (HPLC) of chiral sorbent to split.

Additionally, the compounds of this invention includes their salt, it is also possible to obtain with their hydrate forms, or include them Other solvent used by crystallization.The compounds of this invention can inherently or be formed molten by design with acceptable solvent (including water) Agent compound；Therefore, the present invention includes solvation or non-solvated form.Term " solvate " represents the compounds of this invention The molecular complex of (including its officinal salt) and one or more solvent molecules.These solvent molecules are conventional in pharmaceutical field Those, it is known that it is harmless to experimenter, such as water, ethanol etc..Term " hydrate " represents wherein solvent molecule It it is the complex of water.

The compounds of this invention includes that its salt, hydrate and solvate can be inherently or by design forming polymorphics Thing.

Some compound of formula I can exist with different tautomeric forms.Tautomerism is people in the art Known to Yuan, and technical staff will easily differentiate which group can occur tautomerism to form different tautomerism shapes Formula.The present invention includes all tautomeric forms of compound of formula I.Particularly, the compounds of this invention can be sent out in the following manner Raw tautomerism:

Two kinds of tautomeric forms are considered within the scope of the claims.Therefore, a kind of specific tautomerism is mentioned Form is intended to include any and all optional tautomeric forms.

The compounds of this invention can exist with non-solvated and solvation form.Term used herein " solvate " is retouched Having stated the molecular complex comprising the compounds of this invention and one or more acceptable solvent molecules, described solvent molecule is e.g. Ethanol.When described solvent is water, use term " hydrate ".

Synthesis

Formula (I) compound can such as use the reaction described in described below and embodiment and technology to prepare.These Reaction and can be suitable in the solvent of conversion to be achieved carrying out being suitable for agents useful for same and material.Organic synthesis field Skilled artisan understands that, the functional group being present on molecule should be consistent with the conversion proposed.Sometimes this needs to judge To change the order of synthesis step or to select a kind of to be better than alternative ad hoc approach flow chart to obtain required of the present inventionization Compound.

Multiple substituent group on the following synthetic intermediate shown in reacting flow chart and end-product is in art technology Personnel think can be presented in the protection group being suitable for fully be modified in the case of needs, or can be can lead to subsequently Cross method well-known to those skilled in the art and be modified to the precursor forms existence of its final form.Substituent group can also closed The different phase of one-tenth order is added or is added after synthesis order completes.In many cases, conventional functional group behaviour Make to can be used for one intermediate changes into another kind of intermediate, or one formula (I) compound is changed into another kind of formula (I) change Compound.The example of this generic operation has: ester or ketone are changed into alcohol；Ester is changed into ketone；The mutual conversion of ester, acid and amide；Alcohol Alkylation with amine, acylated and sulfonylation etc..Popular response such as alkylation, acylated, halogenation or oxidation interpolation can also be used to take Dai Ji.This generic operation is well-known in the art, and many reference workss summarize program and the method for this generic operation.Give Go out the main literature of other organic synthesis converted conventional in many functional group manipulations and organic synthesis field Some reference workss of example and reference are March ' s Organic Chemistry, the 5th edition, Wiley and Chichester compiles Collect (2001)；Comprehensive Organic Transformations, Larock edits, VCH (1989)； (series is compiled for Comprehensive Organic Functional Group Transformations, Katritzky et al. Volume), Pergamon (1995)；(series is compiled with Comprehensive Organic Synthesis, Trost and Fleming Volume), Pergamon (1991).Also it is recognized that in any route of synthesis, another kind of main consideration is in plan this area The protection group of the reactive functionality that careful selection is present in compound of the present invention for protection.Same point can be selected Multiple protection groups in son, in order to these protection groups each can be in the case of other protection group in not removing same a part Being removed and same reactions steps maybe can be used to remove several protection groups, this depends on required result.For the operation people undergone training It is Greene and Wuts that member describes authority's report of many alternative selections, Protective Groups in Organic Synthesis,Wiley and Sons(1999)。

Generally, compound of formula I can be synthesized by the route described in flow chart 1 and embodiment.

When A is nitrogen and B is CH, pyridinyl moieties can be according to generalized flowsheet Fig. 1 synthesis shown below.

Flow chart 1

By using relevant initial compounds, similar synthetic method may be used for the compounds of this invention, and wherein A is CH And B is nitrogen.

When A and B is nitrogen, pyrazine part can be according to the 2-in-1 one-tenth of generalized flow chart shown below.

Flow chart 2

When A and B is CH, phenyl moiety can be according to generalized flowsheet Fig. 3 synthesis shown below.

Flow chart 3

Pyrazoles and triazole part form reaction typically by the amide shown in following generalized flowsheet Fig. 4 and add.

Flow chart 4

R¹Group can be by the cross-coupling reaction of the palladium chtalyst shown in following generalized flowsheet Fig. 5 or by following general Aromatic nucleophilic substitution reaction shown in flow chart 6 and introduce.

Flow chart 5

Flow chart 6

In generalized flow chart above, A, B, D, E, R¹And R²As definition the most Anywhere.

The present invention farther includes the variant embodiments of any the inventive method, wherein obtainable in its any stage Midbody product is used as raw material and carries out remaining step, or wherein raw material is to be formed in situ at reaction conditions, or the most anti- Answer component be the salt with them or the pure raw material of optically-active form use.

Can also mutual inversion of phases according to method well known to those skilled in the art, the compounds of this invention and intermediate.

The skilled person will appreciate that general synthetic routes detailed above shows the conventional reaction as converted raw material.Not yet It is provided with concrete reaction condition, but these are well known to those skilled in the art, and in being technical staff's Conventional wisdom Think applicable condition.

Raw material or commercially available or known compound, and the method that can be described by technique of organic chemistry Preparation.

Can be with well known to a person skilled in the art that conventional method formula (I) compound by free form is converted into salt shape Formula, and vice versa.Free or salt form compound can be with hydrate or the solvation containing solvents used for crystallization The form of thing obtains.Formula (I) compound can be reclaimed from reactant mixture by conventional method and is purified.Isomery Body such as stereoisomer can obtain in a usual manner, such as, by fractional crystallization or from (the example of corresponding Asymmetrical substitute Such as optically active) raw material obtained by asymmetric synthesis.

The compound of formula I of free form or salt form shows valuable pharmacological property, such as CFTR control characteristic, example As shown in the in vitro and in vivo test that lower part provides, thus be accordingly used in treatment.

About the regulation of CFTR activity, free or pharmaceutical acceptable salt formula (I) compound can be used for treatment to regulation The disease of CFTR activity response, the disease be particularly benefited from mucosal hydration, such as cystic fibrosis.

Another aspect of the present invention provides the compound of formula I defined the most Anywhere, and it is used as medicine.

Another aspect of the present invention provides compound of formula I, and it is for treating disease or the obstacle of CFTR mediation, especially It is inflammatory or allergic conditions, particularly inflammatory or obstructive airway diseases or mucosal hydration.These diseases include such as capsule Fibrosis, primary ciliary dyskinesia, chronic bronchitis, chronic obstructive pulmonary disease, asthma, respiratory tract infection, pulmonary carcinoma, Xerostomia and keratoconjunctivitis sicca (keratoconjunctivitis sire) or constipation (IBS, IBD, opium sample thing Matter induction).

Another aspect of the present invention provides the free or pharmaceutical acceptable salt defined in any of above embodiment Formula (I) compound is used for treating the disease of CFTR mediation or obstacle, particularly inflammatory or allergic conditions, particularly inflammation in preparation Purposes in the medicine of property or obstructive airway diseases or mucosal hydration.

One embodiment of the invention provides the free or pharmaceutical acceptable salt defined in any of above embodiment Formula (I) compound in preparation for treating the purposes in the medicine of inflammatory or allergic conditions, described disease is selected from capsule Fibrosis, primary ciliary dyskinesia, chronic bronchitis, chronic obstructive pulmonary disease, asthma, respiratory tract infection, pulmonary carcinoma, Xerostomia and keratoconjunctivitis sicca or constipation (induction of IBS, IBD, opioid).

On the other hand, the invention provides the disease treated by CFTR adjustment for the treatment of or the method for obstacle, the method bag Include formula (I) compound of administering therapeutic acceptable amount.In one embodiment, disease or obstacle are inflammatory or anaphylactia Disease, particularly inflammatory or obstructive airway diseases or mucosal hydration.These diseases include that such as cystic fibrosis, constitutional are fine Hair the dyskinesia, chronic bronchitis, chronic obstructive pulmonary disease, asthma, respiratory tract infection, pulmonary carcinoma, xerostomia and be dried Property keratoconjunctivitis or constipation (induction of IBS, IBD, opioid).

Included by the disease of regulation CFTR activity mediation and regulate the disease relevant by the fluid volume of epithelial membrane.Example As, the volume of Airway surface liquid is mucociliary clearance (mucociliary clearance) and the key maintaining lung health Regulation factor.Regulation CFTR activity is by the fluid cumulative on the mucosa side promoting airway epithelia, thus promotes mucociliary clearance And prevent the accumulation in respiratory tissue (including lung airway) of mucus and expectorant.This kind of disease includes respiratory disorder, and such as capsule is fine Dimensionization, primary ciliary dyskinesia, chronic bronchitis, chronic obstructive pulmonary disease (COPD), asthma, respiratory tract infection (acute and chronic；Viral and bacteroidal) and pulmonary carcinoma.By regulation CFTR activity mediation disease also include with By the abnormal fluid regulation of epithelium, the abnormal physiological of protectiveness surface liquid relevant, that may relate on its surface exhales Inhale the disease beyond disease, such as Sjogren syndrome, xerostomia (dry mouth) or keratoconjunctivitis sicca (xerophthalmia).This Outward, the CFTR activity in regulation kidney can be used for promoting diuresis and therefore inducing hypotensive activity.

The treatment of the present invention can be for symptom or preventative.

Asthma includes endogenous (anallergic) asthma and exogenous (allergia) asthma, mild asthma, moderate asthma, weight Degree asthma, bronchitic asthma, the asthma of exercise induced, occupational asthma (occupational asthma) and antibacterial sense The asthma of induction after dye.The treatment of asthma be also understood as including to e.g., less than 4 years old or 5 years old, demonstrate wheezing symptoms and Be diagnosed as maybe can being diagnosed as " wheezy infants " (patient class that a kind of established Major medical is paid close attention to, the most often Be accredited as initial stage or in early days asthma) individual treatment.(for convenience, this special asthma is referred to as " baby Syndrome of stridulating (wheezy infant syndrome) ".)

Preventative effect for the treatment of asthma is by by reducing paresthesia epilepsy, such as acute asthma or the outbreak of bronchoconstriction Frequency and the order of severity, improve pulmonary function or improve airway hyperreactivity and confirm.It can also be right to other by reducing Disease treatment (be used for the most when a symptom occurs or be intended to the treatment for limiting or stop paresthesia epilepsy), such as antiinflammatory are (such as cortex Steroid) or bronchiectasic demand prove.The prevention benefit of asthma is probably spy for easily there is the individuality of " morning dipping " The most obvious.The Asthma Syndrome that " morning dipping " is well recognized as, is common in the asthma of significant proportion, and it is characterized in that such as Morning, time (i.e. generally apart from the time that any property treating asthma of suiting the medicine to the illness used before the is the most remote) asthma of about 4-6 point was sent out Make.

Chronic obstructive pulmonary disease includes chronic bronchitis or associated dyspnea, emphysema and other medicine Thing treatment, the airway hyperreactivity that particularly other Drug therapy sucked is caused deteriorate.Present disclosure additionally applies for treatment to appoint What type or bronchitis of origin, including such as acute bronchitis, arachidic bronchitis, Catarrhal bronchus Inflammation, croupous bronchitis, chronic or phthinoid bronchitis.

Xerophthalmia is characterised by that tear produces and reduces and tear film lipid, protein and mucin abnormal distribution.Xerophthalmia is deposited In many reasons, some of which includes age, laser eye surgery, arthritis, medication, chemicals/thermal burn, allergia and disease Sick such as cystic fibrosis and Sjogren syndrome.Increase and can promote that fluid is from corneal epithelial cell by the anion secretion of CFTR With circumocular eccrine transport to increase cornea aquation.This contributes to alleviating the symptom relevant to xerophthalmia.

Sjogren syndrome is a kind of autoimmune disease, wherein the gland producing moisture content in immune system attack body Body, including eye, mouth, skin, respiratory tissue, liver, vagina and intestinal.Symptom includes xerophthalmia, dry mouth and vagina drying and lung disease Sick.Rheumatoid arthritis, systemic lupus erythematosus (sle), systemic sclerosis and the polymyositis that this disease is still correlated with (polymypositis)/dermatomyositis.Think that protein import defect causes this disease, limited to its therapeutic choice.CFTR lives Property regulator can the multiple organ encroached on by this disease of aquation and contribute to alleviating relevant symptom.

According to above-mentioned, as on the other hand present invention also offers the disease treated regulation CFTR activity response, such as To regulation through the relevant disease of the fluid volume of epithelial cell membrane, the particularly method of obstructive airway diseases, the method bag Include and use formula (I) compound of free form or pharmaceutical acceptable salt to the individuality needed, particularly individual human.

On the other hand, the invention provides formula (I) compound of free form or pharmaceutical acceptable salt, it is right that it is used for treating The disease of regulation CFTR activity response, such as, to regulation through the relevant disease of the fluid volume of epithelial cell membrane, particularly hinder Plug property airway disorders, such as cystic fibrosis and COPD.

On the other hand, the invention provides formula (I) compound of free form or pharmaceutical acceptable salt, be used for for preparation Treat the disease to regulation CFTR activity response, such as to regulation through the relevant disease of the fluid volume of epithelial cell membrane, spy It it not the medicine of obstructive airway diseases, such as cystic fibrosis and COPD.

CFTR active regulator can be tested by measuring the motion of chloride ion in applicable analysis based on cell Fitness as the treatment of the disease having benefited from mucosal hydration.Such as, endogenous expression CFTR or be modified to process LAN The epithelial cell that is unicellular or that converge of CFTR can be used to use electrophysiological technique or ion stream research evaluation channel function.Ginseng See Hirsh et al., J Pharm Exp Ther (2004)；Moody et al., Am J Physiol Cell Physiol (2005) Described in method.

CFTR active regulator, is alternatively arranged as co-therapeutic agent and other medicines such as anti-inflammatory agent, a gas including formula (I) compound Enlargement of pipe agent, hydryllin or anti-tussive agents are applied in combination, particularly at cystic fibrosis or obstructive or airway inflammatory disease In the treatment of (such as those described above), such as this kind of medicine therapeutic activity reinforcing agent or as reduce this The required dosage of class medicine or the means of potential side effect.

The compounds of this invention can with one or more other therapeutic agents simultaneously or before or after use.Chemical combination of the present invention Thing can be administered alone, used by way of using or with other activating agent in identical pharmaceutical composition one by identical or different Rise and use.

In one embodiment, the invention provides the product comprising formula (I) compound and at least one other therapeutic agent Product, they are as combination preparation in the treatment simultaneously, separately or use successively.In one embodiment, treatment is treatment The disease of CFTR mediation or disease.The product provided as combination preparation comprises containing the formula (I) in same medicine compositions The compositions of compound and other therapeutic agent, or formula (I) compound containing divided mode (such as kit form) and other control Treat the compositions of agent.

In one embodiment, the invention provides and comprise formula (I) compound and the drug regimen of other therapeutic agent Thing.Optionally, pharmaceutical composition can comprise above-mentioned pharmaceutically acceptable excipient.

In one embodiment, the invention provides the medicine box comprising the separate pharmaceutical composition of two or more, extremely Few a kind of pharmaceutical composition comprises formula (I) compound.In one embodiment, medicine box comprises the described compositions of independent preservation Device, such as container, separate bottle or separate foil bag.The example of this medicine box is blister package, is commonly used for packaging Tablet, capsule etc..

The medicine box of the present invention may be used for using different dosage forms, the most oral and parenteral, in different administrations Separate compositions is used or for being gradually increased different compositions antagonism another kinds in interval.In order to increase compliance, this Bright medicine box generally comprises uses description.

In the combined therapy of the present invention, the compounds of this invention and other therapeutic agent can be by identical or different productions Producer manufactures and/or preparation.And, the compounds of this invention and other therapeutic agent can be integrated in a combined therapy: (i) exists Before combination product delivers to doctor (such as in the case of comprising the medicine box of the compounds of this invention and other therapeutic agent)；(ii) Doctor oneself (or under doctor instructs) is soon；(iii) patient oneself, such as, using the compounds of this invention successively During other therapeutic agent.

Therefore, the invention provides formula (I) compound purposes in the disease or disease for the treatment of CFTR mediation, wherein make Standby medicine is for using together with other therapeutic agent.Present invention also offers other therapeutic agent in treatment CFTR mediation Purposes in disease or disease, its Chinese medicine is used together with formula (I) compound.

Present invention also offers formula (I) compound, it, for treating disease or the method for disease of CFTR mediation, is wherein made Standby formula (I) compound is for using together with other therapeutic agent.The present invention also provides for other therapeutic agent, and it is used for treating The disease of CFTR mediation or the method for disease, other therapeutic agent wherein prepared is used together with formula (I) compound.The present invention is also Provide formula (I) compound, its for treat CFTR mediation disease or the method for disease, its Chinese style (I) compound with additionally Therapeutic agent use together.Present invention also offers other therapeutic agent, it is for treating disease or the disease of CFTR mediation Method, wherein other therapeutic agent is used together with formula (I) compound.

Present invention also offers formula (I) compound purposes in the disease or disease for the treatment of CFTR mediation, wherein patient Previously with other therapeutic agent treats (such as in 24 hours).Present invention also offers other therapeutic agent in treatment Purposes in the disease of CFTR mediation or disease, wherein patient previously (such as in 24 hours) controls with formula (I) compound Treat.

Accordingly, as another aspect, the present invention includes CFTR active regulator and penetrating agent (saline of height, dextrorotation Sugar acid anhydride, mannitol, xylitol), ENaC blocker, antiinflammatory, bronchodilator, hydryllin, anti-tussive agents, antibiotic and/ Or the combination of deoxyribonuclease medicine, wherein CFTR active regulator and other medicine can be at identical or different medicines In compositions.

The antibiotic being suitable for includes macrolide antibiotic, such as tobramycin (TOBI^TM)。

The deoxyribonuclease medicine being suitable for includes Dornase Alfa (Pulmozyme^TM)-highly purified recombined human takes off The solution of oxygen ribonuclease I (rhDNase), its selective splitting DNA.Dornase Alfa is used for treating cystic fibrosis.

CFTR active regulator with other available combination of anti-inflammatory agent is and chemokine receptor anagonists, such as CCR- 1, CCR-2, CCR-3, CCR-4, CCR-5, CCR-6, CCR-7, CCR-8, CCR-9 and CCR10, CXCR1, CXCR2, CXCR3, CXCR4, CXCR5, particularly CCR-5 antagonist, as Schering-Plough antagonist SC-351125, SCH-55700 and The antagonist of SCH-D, Takeda such as N-[[4-[[[6,7-dihydro-2-(4-methylphenyl)-5H-benzo-cycloheptene-8-base] Carbonyl] amino] phenyl]-methyl] tetrahydrochysene-N, N-dimethyl-2H-pyrans-4-ammonium chloride (TAK-770) and USP6,166, (particularly right is wanted for 037 (particularly claim 18 and 19), WO00/66558 (particularly claim 8), WO00/66559 Ask 9), the combination of the CCR-5 antagonist described in WO04/018425 and WO04/026873.

The anti-inflammatory agent being suitable for includes steroid, particularly glucocorticoid, such as budesonide, beclometasone In (beclamethasone dipropionate), fluticasone propionate, ciclesonide or momestasone furoate, or following documents Described steroid: WO02/88167, WO02/12266, WO02/100879, WO02/00679 (particularly embodiment 3,11, 14, those of 17,19,26,34,37,39,51,60,67,72,73,90,99 and 101), WO03/35668, WO03/48181, WO03/62259, WO03/64445, WO03/72592, WO04/39827 and WO04/66920；On-steroidal glucocorticoid is subject to Body agonist, such as those described in following documents: DE10261874, WO00/00531, WO02/10143, WO03/ 82280、WO03/82787、WO03/86294、WO03/104195、WO03/101932、WO04/05229、WO04/18429、 WO04/19935 and WO04/26248；LTD4 antagonist, such as montelukast and zafirlukast；PDE4 inhibitor, the most western Lip river Department spy (GlaxoSmithKline), roflumilast (Byk Gulden), V-11294A (Napp), BAY19-8004 (Bayer), SCH-351591 (Schering-Plough), arofylline (Almirall Prodesfarma), PD189659/ PD168787(Parke-Davis)、AWD-12-281(Asta Medica)、CDC-801(Celgene)、SelCID(TM)CC- 10004(Celgene)、VM554/UM565(Vernalis)、T-440(Tanabe)、KW-4490(Kyowa Hakko Kogyo) With at WO92/19594, WO93/19749, WO93/19750, WO93/19751, WO98/18796, WO99/16766, WO01/ 13953、WO03/104204、WO03/104205、WO03/39544、WO04/000814、WO04/000839、WO04/005258、 WO04/018450、WO04/018451、WO04/018457、WO04/018465、WO04/018431、WO04/018449、WO04/ 018450、WO04/018451、WO04/018457、WO04/018465、WO04/019944、WO04/019945、WO04/ Those disclosed in 045607 and WO04/037805；Those described in adenosine A 2B receptor antagonist, such as WO02/42298； And beta-2-adrenoceptor agonist, such as albuterol (salbutamol), orciprenaline, terbutaline, salmaterol, non-promise Special sieve, procaterol and particularly formula (I) chemical combination in formoterol, carmoterol and officinal salt, and WO00/75114 Thing (free form or salt form or solvate forms), is incorporated herein by reference the document, preferably the change of embodiment Compound, particularly following formula: compound:

Corresponding to QAB-149 and officinal salt thereof, and formula (I) compound (free form or the salt shape of WO04/16601 Formula or solvate forms), also the compound in documents below: EP1440966, JP05025045, WO93/18007, WO99/64035、USP2002/0055651、WO01/42193、WO01/83462、WO02/66422、WO02/70490、WO02/ 76933、WO03/24439、WO03/42160、WO03/42164、WO03/72539、WO03/91204、WO03/99764、WO04/ 16578、WO04/22547、WO04/32921、WO04/33412、WO04/37768、WO04/37773、WO04/37807、WO04/ 39762, WO04/39766, WO04/45618, WO04/46083, WO04/80964, WO04/108765 and WO04/108676.

The bronchodilator being suitable for includes anticholinergic or muscarine antagonist, particularly ipratropium bromide, oxygen torr bromine Ammonium, tiotropium salt and CHF4226 (Chiesi) and glycopyrronium bromide, also those described in documents below: EP424021, USP3,714,357、USP5,171,744、WO01/04118、WO02/00652、WO02/51841、WO02/53564、WO03/ 00840, WO03/33495, WO03/53966, WO03/87094, WO04/018422 and WO04/05285.

The dual anti-inflammatory and the bronchodilator that are suitable for include that dual beta-2-adrenoceptor agonist/muscarine is short of money Anti-agent, such as those disclosed in USP2004/0167167, WO04/74246 and WO04/74812.

The antihistaminic being suitable for includes cetirizine hydrochloride, acetaminophen, clemastine fumarate, promethazine, chlorine Lei Tading, Desloratadine, diphenhydramine and fexofenadine hydrochloride, acrivastine (activastine), astemizole, nitrogenSTING, ebastine, epinastine, mizolastine and tefenadine (tefenadine), and JP2004107299, Those disclosed in WO03/099807 and WO04/026841.

As another aspect, the present invention includes the combination of CFTR active regulator and CFTR correction agent (corrector), Wherein CFTR active regulator and CFTR correction agent can be in identical or different pharmaceutical compositions.The CFTR correction agent being suitable for Including VX-809

With

VX-661

On the other hand, the invention provides the pharmaceutical composition comprising the compounds of this invention and pharmaceutically suitable carrier.

Pharmaceutical composition can be formulated for particularly using by way of, the most Orally administered, parenteral administration and rectum are executed With etc..Additionally, the pharmaceutical composition of the present invention can make solid form (include but not limited to capsule, tablet, pill, Granule, powder or suppository), or liquid form (including but not limited to solution, suspensoid or Emulsion).Pharmaceutical composition can enter The pharmaceutical operations that row is conventional, such as sterilizing and/or inert diluent, lubricant or buffer agent and the assistant of routine can be comprised Agent, such as preservative, stabilizer, wetting agent, emulsifying agent and buffer agent etc..

Generally, pharmaceutical composition is tablet or gelatine capsule agent, and it comprises active component, and

A) diluent, such as lactose, dextrose, sucrose, mannitol, sorbitol, cellulose and/or glycine；

B) lubricant, such as silicon dioxide, Pulvis Talci, stearic acid, its magnesium salt or calcium salt and/or Polyethylene Glycol；For sheet Agent can also comprise

C) binding agent, such as aluminium-magnesium silicate, gelatinized corn starch, gelatin, Tragacanth, methylcellulose, sodium carboxymethyl cellulose and/ Or polyvinylpyrrolidone；If needing to comprise

D) disintegrating agent, such as starch, agar, alginic acid or its sodium salt, or effervescent mixture；And/or

E) absorbent, coloring agent, correctives and sweeting agent.

Tablet can carry out film coating or enteric coating according to methods known in the art.

Being applicable to the compounds of this invention that Orally administered compositions comprises effective dose, it can be tablet, lozenge, aqueous Or Oil suspensions, dispersible powder or granule, Emulsion, hard or soft capsule or syrup or elixirs.It is intended for mouth The compositions that clothes use can be prepared according to any method known to pharmaceutical composition of preparing that this area uses, and this type of Compositions can be pharmaceutically smart with offer selected from the composition of sweeting agent, correctives, coloring agent and preservative containing one or more Cause and agreeable to the taste preparation.Tablet can prepare the nontoxic pharmaceutically useful of tablet containing active component and being applicable to of being mixed with Excipient.These excipient include: such as inert diluent, such as calcium carbonate, sodium carbonate, lactose, calcium phosphate or sodium phosphate；System Granule and disintegrating agent, such as corn starch or alginic acid；Binding agent, such as starch, gelatin or arabic gum；And lubricant, example Such as magnesium stearate, stearic acid or Pulvis Talci.Tablet is uncoated, or by known technology coating to delay it in the gastrointestinal tract Disintegrate and absorption, thus provide continuous action in a long time.It is for instance possible to use time delay material is such as single stearic Acid glyceride or distearin.Formulations for oral use can be hard-gelatin capsules, wherein active component with Inert solid diluent (such as calcium carbonate, calcium phosphate or Kaolin) mixes；Or be Gelseal, wherein active component with Water or oil medium (such as Oleum Arachidis hypogaeae semen, liquid paraffin or olive oil) mixing.

Some Injectable composition is isotonic aqueous solution or suspension, and suppository is advantageously from fat milk or suspension system Standby.Described compositions can be aseptic and/or containing adjuvant, such as preservative, stabilizer, wetting agent or emulsifying agent, solution Accelerator, the salt of regulation osmotic pressure and/or buffer agent.Additionally, they can also be containing the upper valuable material of other treatment.Institute State compositions can prepare according to conventional mixing, granulation or coating method respectively, and containing about 0.1-75% or containing about The active component of 1-50%.

It is applicable to the compositions the compounds of this invention that comprises effective dose of percutaneous application and applicable carrier.Be applicable to through The carrier that skin delivers includes contributing to the absorbable pharmacologically acceptable solvent through Host Skin.Such as, transdermal dress Be set to the form of binder, its comprise Backing Member, containing described compound and optionally the reservoir of carrier, optional rate controlling barrier with Within a period of time extended, with controlled and predetermined speed, compound is transferred to Host Skin and secures the appliance to skin Instrument on skin.

Be applicable to the compositions of topical application (being such as applied to skin and eye) include aqueous pharmaceutical, suspensoid, ointment, Ointment, gel or spray agent, such as delivering through aerosol etc..This local delivery system is particularly suitable for skin Application, such as, be used for treating skin carcinoma, such as preventive use in sunscreen cream, washing liquid, spray etc..Therefore they are the suitableeest Use together in local, including cosmetic formulations well known in the art.This type of preparation can contain solubilizing agent, stabilizer, tension force enhancing Agent, buffer agent and preservative.

Topical application used herein can also relate to suck or intranasal application.They can be easily with dried powder Form (single form, form of mixtures, such as with the drying composite of lactose, or mixed type component particles, such as with phospholipid Mixed type component particles) delivered by dry powder inhaler, or with aerosol spray presentation use or do not use suitable Delivered by pressurizing vessel, pump, spraying, aerosol apparatus or nebulizer under the propellant closed.

Invention further provides the anhydrous pharmaceutical composition mediating recipe comprising the compounds of this invention as active component Type, because water may promote the degraded of some compound.

The anhydrous pharmaceutical composition of the present invention and dosage form can use the anhydrous or composition of low water content and at low moisture Or prepare under low-moisture conditions.Anhydrous pharmaceutical composition can be produced and store so that keeping its anhydrous nature.Therefore, adopt Prevent the material being exposed to water from anhydrous composition being packed so that they may be packaged in applicable rule with known Determine in medicine box.The example of the packaging being suitable for includes but not limited to seal paper tinsel, plastics, unit-dose container (such as bottle), bubble-cap Packaging and band are packed.

Invention further provides and comprise one or more decomposition reducing the compounds of this invention as active component The pharmaceutical composition of the composition of speed and dosage form.The herein referred to as specific examples of such components of " stabilizer " includes but not limited to antioxidation Agent, such as ascorbic acid, pH buffer agent or salt buffer agent etc..

When compositions includes aerosol formulation, it preferably comprises such as hydrogen-fluoro-alkane (HFA) propellant, such as HFA134a or HFA227 or their mixture, and one or more cosolvent known in the art, such as second can be contained Alcohol (at most 20% weight), and/or one or more surfactants, such as oleic acid or Sorbitan Trioleate, and/or a kind of or Multiple filler, such as lactose.When compositions includes dry powder formulations, it preferably comprises such as has at most 10 micron grain sizes Formula (I) compound, optionally the diluent or carrier such as lactose containing the distribution of required particle diameter, and contribute to preventing properties of product because of wet The compound that gas deteriorates, such as magnesium stearate.When compositions includes atomization preparation, it preferably comprises and is such as dissolved in or is suspended in Formula (I) compound in vehicle, described vehicle includes water, cosolvent such as ethanol or propylene glycol and can be to live in surface The stabilizer of property agent.

Other aspects of the present invention include:

(a) inhalable form, such as aerosol or other nebulizable composition or inhalable particles such as Micronised form Formula (I) compound；

B () comprises the inhalable drug of formula (I) compound of inhalable form；

C () comprises formula (I) compound and the drug products of suction apparatus of inhalable form；With

What d () comprised the compound of formula I of inhalable form can suction apparatus.

The pharmaceutical composition of the present invention or combination can be the lists of about 1-1000mg active component individual for about 50-70kg Position dosage, or about 1-500mg or about 1-250mg or about 1-150mg or about 0.5-100mg or about 1-50mg active component.Generally, Using for sucking, the daily dose being suitable for is 0.005-10mg level, and for Orally administered, the daily dose being suitable for is 0.05- 100mg level.The therapeutically effective amount of compound, its pharmaceutical composition or combination depends on the type of individual treated, body weight, year Age and individual condition, obstacle or disease or its order of severity.The doctor of ordinary skill, clinician or veterinary can be the most true The effective dose of the fixed every kind of active component preventing, treat or suppressing needed for obstacle or disease process.

Above-mentioned dosage character is advantageously to use mammal, the such as organ of mice, rat, Canis familiaris L., monkey or separation, tissue Measure in testing in vitro and in vivo with its prepared product.The compounds of this invention can with the form of solution, such as aqueous pharmaceutical, External application, and such as use as in suspension or aqueous pharmaceutical enteral, non-bowel, advantageously intravenous body.External dose Weight range can be about 10^-3Molar concentration is to 10^-9Molar concentration.Interior therapeutic effective dose scope can depend on using by way of, Between about 0.1-500mg/kg, or between about 1-100mg/kg.

Term used herein " pharmaceutically suitable carrier " includes any and all of solvent, dispersant media, coating, surface Activating agent, antioxidant, preservative (such as antibacterial, antifungal), isotonic agent, absorption delaying agent, salt, preservative, medicine Stabilizer, binding agent, excipient, disintegrating agent, lubricant, sweeting agent, correctives, dyestuff etc. and combinations thereof, it is for this area For technical staff be known (for example, see Remington ' s Pharmaceutical Sciences, the 18th edition, Mack Printing Company, 1990, the 1289-1329 page).Except any conventional carrier incompatible with active component, its Purposes in treatment or pharmaceutical composition is considered.

The compounds of this invention of term " therapeutically effective amount " refers to make individual this of biology or medicinal response of producing The decline of the amount of bright compound, such as enzyme or protein active or suppression, or remission, alleviate disease, slow down or postpone disease Sick process, or prevention disease etc..In a non-limiting embodiment, term " therapeutically effective amount " refers to chemical combination of the present invention The amount of thing, when it is applied to individuality, (1) can at least partly alleviate, suppresses, prevents and/or improve disease or obstacle effectively Or disease, described disease or obstacle or disease are (i) CFTR mediations, or (ii) is relevant to CFTR activity, or (iii) is special Levy the activity (normal or abnormal) being CFTR；Or (2) reduce or suppress the activity of CFTR；Or (3) reduce or suppress CFTR's Express.In another non-limiting embodiment, term " therapeutically effective amount " mean when be applied to cell or tissue or non-carefully When born of the same parents' biomaterial or medium, the amount of the compounds of this invention the most at least partly reduces or suppresses the activity of CFTR；Or at least Part reduces or the expression of suppression CFTR.

Term used herein " individual " refers to animal.Generally animal is mammal.Individuality can also refer to that such as spirit is long Class (such as people, sex), cattle, sheep, goat, horse, Canis familiaris L., cat, rabbit, rat, mice, fish, bird etc..Some embodiment party In case, individuality is primates.In other embodiments, individuality is people.

Term used herein " suppresses " represent minimizing or limit given disease, symptom or obstacle or disease, or biological Being remarkably decreased of the Baseline activity of activity or process.

In one embodiment, any disease " treated " in term used herein or obstacle represents and improves disease or obstacle (i.e. slowing down or stop or reduce the development of disease or at least its a kind of clinical symptoms).In another embodiment, " treat " Represent and alleviate or improve at least one body parameter, can not be by those of patient identification including possibility.In another embodiment In, " treatment " refers to (such as stablize recognizable symptom) on health or regulates disease physiologically (such as stablizing body parameter) Sick or obstacle, or the two all has.In another embodiment, " treat " expression and prevent or delay the outbreak of disease or obstacle Or develop or process.

If individual physical ability used herein in biology, medical science or benefits from this type for the treatment of in terms of quality of life, then it is assumed that should Individual " needs " described treatment.

Herein, unless otherwise indicated or contradiction clear and definite with context, the term " that otherwise the context of the invention uses Individual ", " a kind of ", " being somebody's turn to do " should be construed to include odd number and plural form with similar term (the most in the claims).

Medicinal usage and analysis

Formula (I) compound and officinal salt (hereinafter also referred to " activating agent of the present invention ") thereof can be used as medicine.Especially, Described compound is applicable CFTR active regulator and can test in following assay.

Transmembrane potential algoscopy

By measuring transmembrane potential, CFTR activity can be carried out quantitatively.The method of the transmembrane potential in measurement biosystem Can make in many ways, including the transmembrane potential algoscopy of electrophysiology and optically-based fluorescence.

Optical membrane potential assay uses electronegative potential measurement dyestuff, such as FLIPR transmembrane potential dyestuff (FMP) (see Baxter DF, Kirk M, Garcia AF, Raimondi A, Holmqvist MH, Flint KK, Bojanic D, Distefano PS,Curtis R,Xie Y.‘A novel membrane potential-sensitive fluorescent Dye improves cell-based assays for ion channels. ' J Biomol Screen.2002 February；7 (1): 79-85), its combined quenching agent when born of the same parents are outer.After cell depolarization, electronegative dyestuff reassign to intracellular every Room, thus discharge from film impermeable quencher, cause fluorescence to increase.This fluorescence changes and may cause because of CFTR activity Transmembrane potential change over direct ratio.Can be by the fluorescence detector such as FLIPR (fluorometric imaging plate reader) of proper distrabution 96 Or 384-hole microtitration plate is monitored the change of fluorescence in real time.

Cell is cultivated:

Transmembrane potential test is carried out with Chinese hamster ovary (CHO) cell of stable expression Δ F508-CFTR passage.By cell In 37 DEG C, at 5%v/v CO₂In, maintain in improvement Eagles culture medium (MEM) under 100% humidity, this culture media supplemented 8%v/v hyclone, 100 μ g/mL methotrexate and 100U/mL penicillin/streptomycin.Cell is made to be grown in 225cm²Group Knit in culture flask.In order to carry out transmembrane potential mensuration, by cell with 40,000 cells/well is seeded in 96 orifice plates so that it is viscous Attached, it is maintained at 26 DEG C and reaches 48 hours so that passage inserts.

Reinforcing agent algoscopy:

Transmembrane potential Screening test method make use of the low chlorine ion (～5mM) containing the outer solution of born of the same parents and two-cocoon feeding to add scheme.The Once adding is the buffer with or without test compound, and adding Forskolin (1-20 μM)-program after 5 minutes has It is beneficial to flow out in response to the maximum chlorine of Δ F508-CFTR activation.The chloride ion of Δ F508-CFTR mediation flows out and causes film to go to pole Change, optionally through FMP dyestuff, it is monitored.

Solution:

Outer solution (mM): 120 gluconic acid Na, the 1.2CaCl of low chlorine born of the same parents₂、3.3KH₂PO₄、0.8K₂HPO₄、1.2MgCl₂、 10.0D-glucose, 20.0HEPES, be adjusted to pH7.4 with NaOH

FMP dyestuff: prepare the outer solution of above-mentioned low chlorine born of the same parents, 10 × final concentration according to the explanation of manufacturer, store with 1mL equal portions In-20 DEG C.

IonWorks Quattro algoscopy:

CFTR activity can be carried out quantitatively in electrophysiology mode to use the patch clamp technique of full cell configuration (Hamill et al. Pflugers Acrhive1981).This algoscopy is directly measured relevant to by the chlorine stream of CFTR passage Electric current, maintains simultaneously or adjusts transmembrane voltage.This algoscopy can use single glass micropipettor or planar array in parallel Measure the CFTR activity from natural or reconstitution cell system.The instrument such as IonWorks of suitably outfit can be used It is fixed that the electric current using planar array in parallel to measure is carried out by Quattro (Molecular Devices) or Qpatch (Sophion) Amount.Quattro system can be measured from unicellular/register hole (HT structure) or the group/hole from 64 cells The CFTR electric current of (Population Patch Clamp PPC) (Finkel A, Wittel A, Yang N, Handran S, Hughes J,Costantin J.‘Population patch clamp improves data consistency and Success rates in the measurement of ionic currents. ' J Biomol Screen.2006 8 Month；11 (5): 488-96).

Cell is cultivated:

IonWorks Quattro is carried out with Chinese hamster ovary (CHO) cell of stable expression Δ F508-CFTR passage Test.By cell in 37 DEG C, at 5%v/v CO₂In, maintain under 100% humidity and supplement 10% (v/v) FCS, 100U/ ML penicillin/streptomycin, 1% (v/v) NEAA, 1mg/mL Zeocin and 500ug/mL HYG D-MEM in.In order to enter Row test, makes cell be grown in 225cm²To closely converging in tissue culture flasks, then in 26 DEG C cultivate 48-72 hour so that Passage inserts.From flask, take out cell, be resuspended in for carrying out immediately in the Recording out cell solution tested or again mixing It is suspended in and supplements in the growth medium of 10%v/v DMSO and be refrigerated to-80 DEG C in case use later with 1-2mL equal portions.

Reinforcing agent algoscopy:

Cell is placed in Quattro system with the density of 1.5-3 million/mL, joins in plane patch array, and Seal so that it is set up 5-10 minute.After evaluating and sealing resistance (generally > 50M Ω), by wearing with 100 μ g/mL amphotericin Bs Hole obtains full cell pathway.By scanning survey base current before apply the compound that voltage obtains from-100 to+100mV. Then the test compound by buffer or with the born of the same parents supplementing 20 μMs of Forskolins outer solution dilution joins plane patch battle array In each hole in 384 holes of row.After incubation step (5-20 minute), again survey by applying voltage from-100 to+100mV Determine compound after current.Difference between current between scanning before compound and after compound defines the effect that CFTR strengthens.

Solution:

Born of the same parents outer solution (ECS): 145mM NaCl, 4mM CsCl, 5mM D-Glucose, 10mMTES, 1mM CaCl₂、1mM MgCl₂, pH7.4NaOH

Intracellular buffer (ICS): 113mM L-Aspartic acid, 113mM CsOH, 27mM CsCl, 1mM NaCl, 1mM MgCl₂、1mM EGTA、10mM TES.It is adjusted to pH7.2 with CsOH.Filter Sterile is used before using.

Ion transport algoscopy:

The another kind of method measuring CFTR function is Ussings room short circuit current measurement.Make transformation or natural epithelium Cell is grown to the monolayer that converges on semi-transparent filter membrane and is clipped between two methyl methacrylate (perspex) blocks.Can With the flow by measurement electric current, the chloride ion flow flowing to opposite side via the side from epithelium of CFTR is carried out quantitatively, with Time transepithelial current potential is maintained 0mV.This be use KCl fill electrode based on agar realize to vise cell monolayer also And measure the flow of electric current.

Cell is cultivated:

32mM NaHCO is being supplemented as growth medium₃, 10%v/v hyclone, 2 mM L-glutaminate, In the F-12 culture medium that the Coon of 100U/mL penicillin, 100 μ g/mL streptomycins and 30 μ g/mL HYGs improves on plastics Cultivate the FRT cell stably expressing Δ F508-CFTR.In order to carry out the test of Ussing room, make cell as polarization epithelial growth Support on insert (500000 cell/inserts in growth medium) in Snapwell permeability, and cultivate 7-9 My god.Every 48 hours and test first 24 hours and fill, carrying out Ussing room, the F-12 growth that fresh Coon improves to insert Culture medium.In order to increase Δ F508CFTR protein expression on cell surface, by plate in 27 DEG C of incubations 48 hours, then carry out Ussing room is tested.

Reinforcing agent algoscopy:

The Fischer of the stable expression people Δ F508-CFTR of use monolayer cultures form on permeable holder Rat thyroid (FRT) epithelial cell.In Ussing room, use short circuit current technology, survey to top the end of from force Cl^-Cl is measured under gradient^-Electric current.In order to measure stable Cl^-Electric current, cultivates 48 hours so that Δ by FRT cell at 27 DEG C F508CFTR inserts plasma membrane.The research of Ussing room is carried out at 27 DEG C equally.Under these conditions, it is possible to use titer and effect are eventually The test compound effect to Δ F508 CFTR electric current is added in point quantitatively accumulation.By compound after adding 10 μMs of Forskolins Top is joined and at the end.The effect of compound is compared with known reinforcing agent such as gensitein.

Solution:

Ringer's solution (mM): 126NaCl, 24NaHCO at the end₃、0.38KH₂PO₄、2.13K₂HPO₄、1MgSO₄、1CaCl₂ With 10 glucoses.

Top ringer's solution (mM): 140 gluconic acid Na, 1MgSO₄、2CaCl₂, 1HCl, 10 glucoses and 24NaHCO₃。

Said determination method test compound can also be used to stimulate Δ F508CFTR to insert the ability of cell membrane.For these Algoscopy, scheme is identical, except for the difference that cell not under low temperature (26 or 27 DEG C) cultivate, but before the assay with testization Compound incubation 12-24 hour together.

In above-mentioned measurement data, the compound of Examples below is generally of Ki value less than 10 μMs.Such as, following enforcement Example compound has a special Ki value: embodiment 5:0.07 μM, embodiment 7:0.01 μM, embodiment 14:0.255 μM, embodiment 18:0.004 μM, embodiment 19:1.55 μM, embodiment 22:0.025 μM, embodiment 24:0.02 μM, embodiment 25.1:3.8nM, Embodiment 25.2:13nM, embodiment 25.3:14nM, embodiment 26.1:30nM, embodiment 26.2:45nM, embodiment 26.3: 0.160 μM and embodiment 27:19nM.

Unless otherwise indicated or otherwise clearly contradicted, otherwise all methods described herein can be with any applicable Order is carried out.The use of provided herein any and all embodiment or exemplary language (such as " such as ") is only for more all right The scope of the present invention that the bright present invention rather than restriction require.

The present invention is to be illustrated by the following example.

Embodiment

General conditions:

The LC-MS system using electron spray ionisation is run mass spectrum.These are Agilent1100HPLC/Micromass Platform Mass Spectrometer combination or the Waters Acquity with SQD Mass Spectrometer UPLC。[M+H]⁺Refer to list-isotopic molecule amount.

The open entrance Bruker AVANCE400NMR spectrophotometer using ICON-NMR runs H NMR spectroscopy.? 298K measures spectrum and uses solvent peak as reference.

The following example is intended to for illustrating the present invention, but should not be understood as limitation of the present invention.Temperature is With degree Celsius be given.Without mentioning other condition, the most all of evaporation is the most under reduced pressure carried out, preferably about Carry out under 15mm Hg to 100mm Hg (=20-133mbar).The structure of end-product, intermediate and raw material is to pass through standard scores Analysis method, such as microanalysis and spectral signature, such as MS, IR and NMR confirm.Abbreviation used is conventional in this area Those.If it is not defined, then term has its implication accepted extensively.

Abbreviation:

App is apparent

ATP ATP

BINAP raceme 2,2 '-bis-(diphenylphosphines)-1,1 '-dinaphthalene

9-BBN 9-boron bicyclo-[3.3.1] nonane

BOC tert-butyl carboxy

Br width

BSA bovine serum albumin

D is bimodal

Dd double doublet

DCM dichloromethane

DIEA diethyl isopropyl amine

DIPEA diisopropyl ethyl amine

DMF N,N-dimethylformamide

DMSO dimethyl sulfoxide

DTT dithiothreitol, DTT

EDTA ethylenediaminetetraacetic acid

ESI electron spray ionisation

EDCI 1-ethyl-3-(3 '-dimethylamino-propyl) carbodiimide

EtOAc ethyl acetate

FCC flash column chromatography

H hour

HATU 2-(7-azepine-1H-benzotriazole-1-base)-1,1,3,3-tetramethylureaHexafluorophosphate

HBTU 1-[double (dimethylamino) methylene]-1H-benzotriazoleHexafluorophosphate (1-) 3-oxide

HOBt 1-hydroxyl-7-azepine benzotriazole

HPLC high pressure lipuid chromatography (HPLC)

IR infrared spectrometry

LCMS liquid chromatography and mass spectrography

MeOH methanol

MEMCl 2-methoxvethoxvmethvl chlorine

MS mass spectrography

MW microwave

M multiplet

Min minute

ML milliliter

M/z mass-to-charge ratio

NMR nuclear magnetic resonance, NMR

Ppm million/

PyBOP benzotriazole-1-base epoxide tripyrrole alkane subbase phosphorusHexafluorophosphate

PS polymer is supported

PPTS pyridineTosilate

PEAX PE-anion exchange be (such as Biotage'sPE-AX post)

Rac racemic modification

RT room temperature

S is unimodal

SCX-2 strong cation exchange be (such as Biotage'sSCX-2 post)

T triplet

TEA triethylamine

TFA trifluoroacetic acid

THF oxolane

Tris HCl amino three (hydroxymethyl) methane hydrochloride salt

With reference to the following examples, method described herein or other method known in the art is used to synthesize preferred reality Execute the compound of scheme.

In a suitable manner, it is possible to use routine techniques such as precipitates, filter, crystallize, evaporate, distill and chromatography divides From plurality of raw materials, intermediate and compound with purification preferred embodiment.Unless otherwise stated, all raw materials are all obtained from Commercial supplier and the most i.e. using.Salt can be prepared by compound by the operation of known one-tenth salt.

It should be appreciated that the organic compound of preferred embodiment can show tautomerism.Due to this explanation Chemical constitution in book is only capable of one of tautomeric form of expressing possibility, it is, therefore, appreciated that preferred embodiment bag Include any tautomeric form of shown structure.

If not stated otherwise, then analytical type HPLC condition is as follows:

Method 2minLC_v002

Post Waters BEH C1850 × 2.1mm, 1.7 μm

Column temperature 50 DEG C

Eluent A:H₂O, B: methanol, all containing 0.1%TFA

Flow velocity 0.8mL/ minute

0.20 minute 5%B of gradient；5% to 95%B in 1.30 minutes, 0.25 minute 95%B

Method 2minLC_v003

Post Waters BEH C1850 × 2.1mm, 1.7 μm

Column temperature 50 DEG C

Eluent A:H₂O, B: acetonitrile, all containing 0.1%TFA

Flow velocity 0.8mL/ minute

0.20 minute 5%B of gradient；5% to 95%B in 1.30 minutes, 0.25 minute 95%B

The preparation of finalization compound

Particularly preferred the compounds of this invention includes that those shown in table 1 below, preparation method describe later.

Table 1

Embodiment 1

3-amino-6-bromo-5-trifluoromethyl-pyrazine-2-formic acid (5-trifluoromethyl-2H-pyrazole-3-yl)-amide

3-amino-6-bromo-5-trifluoromethyl-pyrazine-2-formic acid (intermediate 2) (100mg, 0.350mmol) to stirring Dry NMP (2mL) solution in add NEt₃(0.097mL, 0.699mmol), is subsequently added 3-(trifluoromethyl)-1H-pyrrole Azoles-5-amine (48.0mg, 0.318mmol).Mixture is stirred 5 minutes at RT, then with HATU (133mg, 0.350mmol) place Reason.The orange solution produced is stirred 10 minutes at RT, then distributes between EtOAc (50mL) and 1M NaOH (30mL).Point From organic moiety and with 1M NaOH (20mL), water (20mL) washing, it is dried (MgSO₄) and be concentrated in vacuo.Silica gel color is provided Spectrometry purification of crude residue, with isohexane/EtOAc (gradient of 0 to 50%EtOAc) eluting, obtains title compound, for yellow Solid.

Embodiment 2-7

These compounds are i.e.

3-amino-6-bromo-5-trifluoromethyl-pyrazine-2-formic acid (5-cyclopropyl-2H-pyrazole-3-yl)-amide (embodiment 2),

3-amino-6-bromo-5-trifluoromethyl-pyrazine-2-formic acid (the 5-tert-butyl group-2H-pyrazole-3-yl)-amide (embodiment 3),

3-amino-6-bromo-5-trifluoromethyl-pyrazine-2-formic acid (5-methyl-2H-pyrazole-3-yl)-amide (embodiment 4),

3-amino-6-bromo-5-trifluoromethylpyridin-2-formic acid (5-trifluoromethyl-2H-pyrazole-3-yl)-amide (is implemented Example 5),

3-amino-6-bromo-5-trifluoromethyl-pyrazine-2-formic acid (2H-pyrazole-3-yl)-amide (embodiment 6),

3-amino-6-bromo-5-trifluoromethylpyridin-2-formic acid (2H-pyrazole-3-yl)-amide (embodiment 7), is similar In 3-amino-6-bromo-5-trifluoromethyl-pyrazine-2-formic acid (5-trifluoromethyl-2H-pyrazole-3-yl)-amide (embodiment 1) by Prepared by the initial compounds (described herein) and the amine that are suitable for.Some reaction microwave radiation is carried out.

Embodiment 8

3-amino-6-pyrrolidin-1-yl-5-trifluoromethyl-pyrazine-2-formic acid (5-trifluoromethyl-2H-pyrazole-3-yl)- Amide

To stirring 3-amino-6-bromo-5-trifluoromethyl-pyrazine-2-formic acid (5-trifluoromethyl-2H-pyrazole-3-yl)- In dry THF (2mL) solution of amide (embodiment 1) (40mg, 0.095mmol) add pyrrolidine (7.89 μ L, 0.095mmol).The mixture microwave radiation produced is heated 35 minutes at 120 DEG C.Reactant mixture is at EtOAc (50mL) And distribute between water (30mL), and separate organic moiety, it is dried (MgSO₄) and be concentrated in vacuo, obtain crude product, for orange Grease.Crude product preparative TLC is purified, uses DCM:1%MeOH eluting, obtain title compound, for orange solids.

Embodiment 9

3-amino-6-morpholine-4-base-5-trifluoromethyl-pyrazine-2-formic acid (2H-pyrazole-3-yl)-amide

3-amino-6-bromo-5-trifluoromethyl-pyrazine-2-formic acid (2H-pyrazole-3-yl)-amide (embodiment 6) will be comprised The mixture of (42mg, 0.120mmol) and morpholine (1mL, 11.3mmol) at 100-140 DEG C with microwave heating 8 hours 15 minutes. Reactant mixture is concentrated in vacuo, and by preparative LC-MS purification residues, obtains title compound, for yellow solid.

Embodiment 10

3-amino-5-trifluoromethyl-pyrazine-2-formic acid (2H-pyrazole-3-yl)-amide

Title compound is analogous to 3-amino-6-bromo-5-trifluoromethyl-pyrazine-2-formic acid (5-trifluoromethyl-2H-pyrrole Azoles-3-base)-amide (embodiment 1) 3-amino-5-trifluoromethyl-pyrazine-2-formic acid (intermediate 6) and 2H-pyrazole-3-yl Prepared by amine.

Embodiment 11

5-[(3-amino-6-bromo-5-trifluoromethyl-pyrazine-2-carbonyl)-amino]-1H-[1,2,4] triazole-3-formic acid

Title compound is analogous to 3-amino-6-bromo-5-trifluoromethyl-pyrazine-2-formic acid (5-trifluoromethyl-2H-pyrrole Azoles-3-base)-amide (embodiment 1) by with 5-amino-1H-[1,2,4] triazole-3-methyl formate replace 3-(trifluoromethyl)- 1H-pyrazoles-5-amine and prepare.

Embodiment 12

3-amino-6-dimethylamino-5-trifluoromethyl-pyrazine-2-formic acid (2H-pyrazole-3-yl)-amide

3-amino-6-bromo-5-trifluoromethyl-pyrazine-2-formic acid (2H-pyrazole-3-yl)-amide (embodiment to stirring 7) dry DMF (5mL) solution of (50mg, 0.142mmol) adds diethanolamine (37.4mg, 0.356mmol).By produce Solution heats 66 hours with microwave radiation at 100 DEG C.Reactant mixture in EtOAc (20mL) and water (20mL) between distribute, And separate each phase.Organic facies saline (10mL) washs, and is dried (MgSO₄) and be concentrated in vacuo, obtain orange solids.Pass through Silica gel chromatography crude product, with isohexane/EtOAc (0 to 50%EtOAc gradient) eluting, obtains title compound, for Huang Color solid.

Embodiment 13

3-amino-6-bromo-5-trifluoromethyl-pyrazine-2-formic acid (2H-[1,2,4] triazole-3-base) amide

Title compound is analogous to 3-amino-6-bromo-5-trifluoromethyl-pyrazine-2-formic acid (5-trifluoromethyl-2H-pyrrole Azoles-3-base)-amide (embodiment 1) by with 1H-[1,2,4] triazole-5-amine replace 3-(trifluoromethyl)-1H-pyrazoles-5-amine And prepare.

Embodiment 14

3-amino-6-chloro-5-trifluoromethyl-pyrazine-2-formic acid (2H-pyrazole-3-yl) amide

Title compound is analogous to 3-amino-6-bromo-5-trifluoromethyl-pyrazine-2-formic acid (5-trifluoromethyl-2H-pyrrole Azoles-3-base)-amide (embodiment 1) 3-amino-6-chloro-5-trifluoromethyl-pyrazine-2-formic acid (intermediate 7) and 2H-pyrazoles- Prepared by 3-base amine.Reaction is within 1 hour, to carry out at 100 DEG C with microwave radiation.

Embodiment 15

3-amino-6-bromo-5-trifluoromethyl-pyrazine-2-formic acid (5-piperidin-1-yl methyl-2H-pyrazole-3-yl)-amide

To comprise 3-amino-6-bromo-5-trifluoromethyl-pyrazine-2-formic acid (intermediate 2) (0.349g, 1.221mmol), 3-(piperidin-1-yl methyl)-1H-pyrazoles-5-amine (intermediate 8) (0.220g, 1.221mmol), HATU (0.464g, 1.221mmol) stir 30 minutes at RT with the N-methylmorpholine (0.241mL, 2.441mmol) mixture in DMF (5mL). Reactant mixture use water (20mL) dilutes, and is filtered under vacuo by the yellow mercury oxide of generation and wash with water (20mL) Wash.The residue produced is loaded in SCX-2 post, uses MeOH eluting, the 7M NH being subsequently used in MeOH₃Eluting.It is concentrated in vacuo Methanol ammonia fraction, and residue EtOAc/ isohexane is ground, obtain title compound.

Embodiment 16

The bromo-N-of 3-amino-6-(1H-pyrazolo [4,3-b] pyrazine-3-base)-5-(trifluoromethyl) pyrazine-2-amide

To comprise 3-amino-6-bromo-5-trifluoromethyl-pyrazine-2-formic acid (intermediate 2) (0.212g, 0.740mmol), 1H-pyrazolo [3,4-b] pyrazine-3-amine (0.10g, 0.740mmol), HATU (0.281g, 0.740mmol) and N-methylmorpholine (0.146mL, 1.480mmol) mixture in DMF (5mL) stirs 45 minutes at RT.By reactant mixture use water (25mL) Dilution, and the orange precipitate produced is filtered under vacuo and washs with water (10mL).Solid is passed through silica gel chromatography Method purification, with isohexane: EtOAc (0 to 100%EtOAc gradient) eluting, obtains title compound, for yellow solid.

Embodiment 17

3-(3-amino-6-bromo-5-(trifluoromethyl) pyrazine-2-amide groups)-4,6-pyrrolin also [3,4-c] pyrazoles-5 (2H)-t-butyl formate

To comprise 3-amino-6-bromo-5-trifluoromethyl-pyrazine-2-formic acid (intermediate 2) (0.102g, 0.357mmol), 3-amino-4,6-pyrrolin also [3,4-c] pyrazoles-5 (2H)-t-butyl formate (0.08g, 0.357mmol), HATU (0.136g, 0.357mmol) and the N-methylmorpholine (0.071mL, 0.713mmol) mixture in DMF (5mL) stir at RT 30 minutes.Reactant mixture use water (15mL) is diluted, and the orange precipitate produced is filtered under vacuo, and use water (10mL) washing.The solid produced by Silica gel chromatography, is washed with isohexane: EtOAc (0 to 100%EtOAc gradient) De-, obtain title compound, for yellow solid.

Embodiment 18

The bromo-N-of 3-amino-6-(1H-pyrazolo [4,3-b] pyrazine-3-base)-5-(trifluoromethyl) picolinamide

To comprise 3-amino-6-bromo-5-trifluoromethylpyridin-2-formic acid (intermediate 3) (0.10g, 0.351mmol), 1H-pyrazolo [3,4-b] pyrazine-3-amine (0.047g, 0.702mmol), HATU (0.133g, 0.351mmol) and N-methyl The quinoline (0.069mL, 0.702mmol) mixture in DMF (5mL) stirs 90 minutes at 75 DEG C.After being cooled to RT, by mixture Dilute with water (30mL), and the orange precipitate produced is filtered under vacuo, and wash with water.Pass through silica gel chromatography The solid that purification produces, with isohexane: EtOAc (0 to 100%EtOAc gradient) eluting, obtains title compound, for orange solid Body.

Embodiment 19

The bromo-N-of 3-amino-6-(4-(pyridine-2-base)-1H-pyrazoles-5-base)-5-(trifluoromethyl) pyrazine-2-amide

To comprise 3-amino-6-bromo-5-trifluoromethyl-pyrazine-2-formic acid (intermediate 2) (0.179g, 0.624mmol), 5-amino-4-(pyridine-2-base)-1H-pyrazoles (0.1g, 0.624mmol), HATU (0.237g, 0.624mmol) and N-methyl The quinoline (0.123mL, 1.249mmol) mixture in DMF (5mL) stirs 2.5 hours at 80 DEG C.After being cooled to RT, will mixing Thing use water (40mL) dilutes, and is filtered under vacuo by the yellow mercury oxide of generation, and washs with water (20mL).To solid Middle addition MeOH (150mL), and ultrasonic suspension, obtain trickle suspension.Suspension is filtered under vacuo, and will Solid MeOH (20mL) drip washing, and be vacuum dried 1.5 hours at 45 DEG C, obtain title compound, for yellow solid.

Embodiment 20

The iodo-N-of 2-amino-5-(1H-pyrazoles-5-base)-4-(trifluoromethyl) benzamide hydrochloride salt

2-amino-5-iodo-4-(Trifluoromethyl)benzoic acid. (intermediate 4) (0.414g, 1.251mmol), 1H-pyrrole will be comprised Azoles-5-amine (0.104g, 1.251mmol) and HATU (0.476g, 1.251mmol) the mixture N-methyl in DMF (5mL) Morpholine (0.124mL, 1.251mmol) processes, and is stirred 1 hour at RT by mixture.Reactant mixture is used 2M NaOH (20mL) dilution, after stirring 5 minutes, product EtOAc (2 × 100mL) extracts.The organic extract merged is merged, is dried (MgSO₄) and be concentrated in vacuo.The solid produced by silica gel column chromatography purification, with isohexane: EtOAc (0 to 100% EtOAc gradient) eluting, obtain yellow oil.By grease dried in vacuum overnight at 45 DEG C, and the white size that will produce Shape thing stirs 2 hours in ether (2mL) solution of 2M HCl, filters, and washs with ether, and 45 DEG C of high vacuum dry, obtains Title compound, for white solid.

Embodiment 21

The bromo-N-of 3-amino-6-(1H-pyrazolo [3,4-b] pyridin-3-yl)-5-(trifluoromethyl) pyrazine-2-amide

To comprise 3-amino-6-bromo-5-trifluoromethyl-pyrazine-2-formic acid (intermediate 2) (0.320g, 1.118mmol), 3-amino-pyrazol also [3,4-b] pyridine (0.150g, 1.118mmol) and HATU (0.425g, 1.118mmol) are in DMF (5mL) Mixture process with N-methylmorpholine (0.111mL, 1.118mmol), and be stirred overnight at 80 DEG C.After being cooled to RT, will Mixture use water (25mL) dilutes, and is filtered under vacuo by the yellow mercury oxide of generation and wash with water.By thick material By Silica gel chromatography, with isohexane: EtOAc (0 to 100%EtOAc gradient) eluting, obtain title compound, for Huang Color solid.

Embodiment 22

The chloro-N-of 3-amino-6-(1H-pyrazoles-5-base)-5-(trifluoromethyl) picolinamide

DMF by 3-amino-6-chloro-5-trifluoromethylpyridin-2-formic acid (intermediate 5) (0.143g, 0.594mmol) (5mL) solution HATU (226mg, 0.594mmol), 1H-pyrazoles-5-amine (74.1mg, 0.892mmol) and 4-methyl morpholine (0.118mL, 1.189mmol) processes.Mixture is stirred 30 minutes at RT, stirs 2 hours at 80 DEG C subsequently, be subsequently cooled to RT.Reactant mixture 2M NaOH (20mL) is diluted, and product EtOAc (2 × 60mL) is extracted.Organic substance is closed And, wash with saline (20mL), through MgSO₄It is dried and is concentrated in vacuo, obtaining brown oil.This grease is dissolved in DCM (2mL), after 5 minutes, there is white depositions in solution.Its vacuum is filtered, washs with DCM, and high vacuum dry.By thing Matter is dissolved in EtOAc (50mL), and successively with 2M NaOH (2 × 20mL), saline (10mL) washing, through MgSO₄It is dried and true Empty concentration, obtains yellow solid.It is dissolved in MeOH (3mL), and loads in 10g SCX-2 post, wash with MeOH (50mL) De-, then it is used in the 7M NH in MeOH (50mL)₃Eluant solution.Merge relevant fraction and be concentrated in vacuo, obtaining titled Compound.

Embodiment 23

5-(3-amino-6-bromo-5-(trifluoromethyl) pyrazine-2-amide groups)-1H-1,2,4-triazole-3-methyl formate

Being dried to 3-amino-6-bromo-5-trifluoromethyl-pyrazine-2-formic acid (intermediate 2) (300mg, 1.049mmol) DCM (10mL) solution adds NEt₃(0.322mL, 2.308mmol), is subsequently added 5-amino-1H-1,2,4-triazole-3-first Acid methyl ester (149mg, 1.049mmol).Stir after 5 minutes at RT, by reactant mixture with PyBOP (546mg, 1.049mmol) Process, and stir 3 hours.Mixture is distributed between DCM (20mL) and water (20mL), shakes and separate.By organic Part use water (20mL) is washed, through MgSO₄It is dried, filters and be concentrated in vacuo, obtain orange.Pass through silica gel chromatography Purification of crude residue, the 0-100%EtOAc eluting being used in isohexane, the 0-35%MeOH eluting being subsequently used in EtOAc, Obtain yellow oil.Grind grease with EtOAc/ isohexane, grind with MeCN/ water subsequently, obtain title compound, for Huang Color solid.

Embodiment 24

3-amino-N-(3-benzyl-1H-1,2,4-triazole-5-base)-6-bromo-5-(trifluoromethyl) pyrazine-2-amide

Title compound is analogous to embodiment 23 by 3-amino-6-bromo-5-trifluoromethyl-pyrazine-2-formic acid (intermediate 2) prepare with 3-benzyl-1H-1,2,4-triazole-5-amine.

Embodiment 25.1

3-amino-6-(4-fluorophenyl)-N-(1H-pyrazole-3-yl)-5-(trifluoromethyl) picolinamide

Step 1:3-amino-6-(4-fluorophenyl)-5-(trifluoromethyl) pyridine carboxylic acid methyl ester

By 3-amino-6-bromo-5-trifluoromethylpyridin-2-methyl formate (intermediate 3D) (200mg, 0.669mmol), 4-flurophenyl boronic acid (94mg, 0.669mmol) and 1,1 '-bis-(diphenylphosphino) ferrocene palladium chloride (Apollo) is suspended in THF (2mL) and 1M Cs₂CO₃(0.667mL) in.By bottle N₂Purging, seals and heats 15 with microwave radiation at 160 DEG C Minute.Reactant mixture is distributed between EtOAc (50mL) and water (50mL).Separate organic moiety and with saline (30mL) Washing, through MgSO₄It is dried, filters and be concentrated in vacuo.By Silica gel chromatography, the 0-80% being used in isohexane EtOAc eluting, obtains title compound；

LCMS:Rt=1.47 minute, [M+H]+315.1；Method 2minLC_v002.

Step 2:3-amino-6-(4-fluorophenyl)-5-(trifluoromethyl) pyridine carboxylic acid

3-amino-6-(4-fluorophenyl)-5-(trifluoromethyl) pyridine carboxylic acid methyl ester (step 1) (6.00mmol) to stirring EtOH (5mL) solution in add 2M NaOH (3mL, 6.00mmol), and by solution RT stir 15 minutes.By produce Mixture use water (10mL) dilutes, and with 1MHCl, pH is adjusted to pH6.Mixture DCM (2 × 10mL) is extracted, and uses The post that is separated separates each phase.The organic extract of merging is concentrated in vacuo, and without being further purified and for next step；

LCMS:Rt=1.42 minute, [M+H]+301.1；Method 2minLC_v002.

Step 3:3-amino-6-(4-fluorophenyl)-N-(1H-pyrazole-3-yl)-5-(trifluoromethyl) picolinamide

Dry NMP by the 3-amino-6-(4-fluorophenyl)-5-(trifluoromethyl) pyridine carboxylic acid (0.167mmol) of stirring (2mL) solution 1H-pyrazoles-3-amine (0.183mmol) processes.After RT stirs, add triethylamine (0.366mmol), and Continue stirring 5 minutes.Add HATU (0.183mmol), and the mixture produced is sealed, and with microwave radiation 100 DEG C heating 1 hour.Mixture is distributed between EtOAc (25mL) and 1M NaOH (25mL).Separate organic moiety, use water (25mL) washing, through MgSO₄It is dried, and is concentrated in vacuo.Orient LCMS purification by quality, use TFA/MeCN/ water elution, To title compound, for tfa salt.Salt is distributed between EtOAc (10mL) and sodium bicarbonate (10mL).Separate organic moiety, By separated post and be concentrated in vacuo, obtain title compound；

LCMS:Rt=1.58 minute, [M+H]+366.2；Method 2minLC_v002.

1H NMR(400MHz,DMSO-d6)δ12.6(1H,s),10.2(1H,s),7.82(1H,s),7.78(1H,s), 7.62(2H,m),7.3-7.4(4H,m),6.72(1H,s)。

The compound (table 2) of following table embodiment be by be similar to the method for embodiment 25 by the initial compounds being suitable for and Prepared by amine.

Table 2

Embodiment 26.1-26.3

The compound (table 3) of following table embodiment is prepared according to following universal method:

3-amino-6-bromo-5-fluoroform is added in the microwave vial containing commercially available boric acid (0.100mL) The 1,4-bis-of base-pyridine-2-formic acid (2H-pyrazole-3-yl)-amide (embodiment 7) (0.100mmol)Alkane (1mL) solution, K₂CO₃(0.300mmol) water (167 μ L) solution, is subsequently added Pd (PPh₃)₂Cl₂(0.7mg, 1.000 μm ol).By mixture Heat 30 minutes with microwave radiation at 160 DEG C.The mixture produced is passed through SCX-2 (1g) post of moistening in advance (MeOH), uses MeOH eluting, the 3M NH being subsequently used in MeOH₃Eluting.Methanol ammonia fraction is concentrated in vacuo, and is vacuum dried.To slightly produce Thing passes through preparative LCMS purification, obtains required product.

Table 3

Embodiment 27

3-amino-5,6-pair-trifluoromethyl-pyrazine-2-formic acid (4-methyl-2H-pyrazole-3-yl)-amide

Title compound is analogous to embodiment 1.0 to be prepared by intermediate 1AA and 4-methyl isophthalic acid H-pyrazoles-5-amine.

LCMS:Rt=1.43 minute；[M+H]+354.9；Method 2minLC_v003

1H NMR (400MHz, DMSO-d6) δ 12.5 (1H, s), 10.2 (1H, s), 8.6-8.8 (1H, wide hump), 8.4- (8.6 1H, wide hump), 7.54 (1H, s), 1.92 (3H, s).

The preparation of intermediate

Intermediate 1

3-amino-5-trifluoromethyl-pyrazine-2-Ethyl formate

Intermediate 1A:Amidino groups (carbamimidoyl)-nitroso-group-ethyl acetate

At 0 DEG C to 5 DEG C, go through in ethanol (152mL, the 0.304mmol) solution of 2M ammonia 30 minutes and add ethyoxyl Carbonyl EA HCl (25g, 0.127mmol).Reaction is stirred 3 hours at this high temperature, the most disposably adds Enter sodium nitrite in aqueous solution (9.63g, 0.139mmol).Add 5N HCl and the pH of mixture is adjusted to pH6.By reactant mixture Place at RT and be stirred overnight.The yellow mercury oxide of formation is filtered under vacuo, washes with water and be dried, obtain title compound Thing；

¹H NMR(400MHz,DMSO-d6)δ10.1(2H,br s),7.6(2H,br s),4.3(2H,q),1.3(3H, t)。

Intermediate 1B:Amino-carbamimidoyl group-ethyl acetate

To ethanol/5M HCl (1:1 ratio, the 250mL) solution of amidino groups-nitroso-group-ethyl acetate (5.5g, 31.4mmol) Middle addition 10%Pd/C (1.3g).Reactant mixture is under low pressure hydrogenated (H_2(g)) 2 evenings.Warp(filtering material) mistake Filter Pd/C, and filter vacuum is concentrated, obtain title compound, for white solid.It is used for next step with crude product.

Intermediate 1:3-amino-5-trifluoromethyl-pyrazine-2-Ethyl formate

Trifluoropropyl keto-aldehyde is added in the mixture of amino-carbamimidoyl group-ethyl acetate (2g, 9.22mmol) and water (50mL) 20% aqueous solution (2.32g, 18.43mmol).(reaction is mixed to add sodium acetate (5.29g, 64.52mmol) in this mixture The pH of compound is pH5).Reactant mixture is placed at RT and is stirred overnight.The precipitate of generation is filtered under vacuo, passes through silicon Glue chromatography purification, with isohexane: EtOAc (0 to 10%EtOAc gradient) eluting, obtains title compound.

¹H NMR(400MHz,DMSO-d6)δ8.4(1H,s),7.8(2H,br s),4.4(2H,q),1.4(3H,t)。

Intermediate 1AA

Double (trifluoromethyl) pyrazine-2-formic acid of 3-amino-5,6-

Step 1:Double (trifluoromethyl) pyrazine-2-Ethyl formate of 3-amino-5,6-

Title compound is analogous to intermediate 1 preparation, by with 1,1, Isosorbide-5-Nitrae, 4,4-hexafluoro butyl-2, and 3-diketone replaces Trifluoropropyl keto-aldehyde；

LCMS Rt=4.72 minute, [M+H]+304.2/326.1；Method 10minLC_v002.

Step 2:Double (trifluoromethyl) pyrazine-2-formic acid of 3-amino-5,6-

To 3-amino-5 of stirring, the EtOH of double (trifluoromethyl) pyrazine-2-Ethyl formate (300mg, 0.990mmol) of 6- (10mL) solution is gone through dropping 2M NaOH (0.495mL, 0.990mmol) in 1 minute.After RT stirs 30 minutes, reaction is mixed Compound is poured in water (30mL), and by adding 1M HCl, pH is adjusted to pH4.Mixture EtOAc (2 × 50mL) is extracted Take, and the organic extract saline (30mL) merged is washed, through MgSO₄(5g) it is dried, filters and be concentrated in vacuo, To title compound, for off-white color crystalline solid；

¹H NMR (400MHz, DMSO-d6) δ 8.6-9.2 (2H, wide hump), 7.8-8.3 (2H, wide hump), 4.4 (2H, q),1.32(3H,t)。

Intermediate 2

3-amino-6-bromo-5-trifluoromethyl-pyrazine-2-formic acid

Intermediate 2A:3-amino-6-bromo-5-trifluoromethyl-pyrazine-2-Ethyl formate

Acetic acid to 3-amino-5-trifluoromethyl-pyrazine-2-Ethyl formate (intermediate 1) (30mg, 0.13mmol) (5mL) solution adds sodium carbonate (15mg, 0.14mmol).The bromine (7 μ L, 0.13mmol) of half amount is added in this mixture Acetic acid (5mL) solution, is subsequently added sodium carbonate (15mg, 0.14mmol).Add the acetic acid solution of remaining bromine, and will reaction Mixture places stirring 2 hours at RT.By mixture dilute with water, and the yellow mercury oxide of generation is filtered under vacuo, Obtain title compound.

Intermediate 2:3-amino-6-bromo-5-trifluoromethyl-pyrazine-2-formic acid

Molten to the ethanol (20mL) of the 3-amino-5-trifluoromethyl-pyrazine-2-Ethyl formate (10g, 31.8mmol) of stirring Liquid adds 2M NaOH (20mL, 31.8mmol).The solution of generation is stirred 5 minutes at RT and pours in water (50mL). Add 1M HCl and pH is adjusted to pH6.The suspension of generation is filtered under vacuo, washs with water (20mL), and be dried, obtain Title compound；

MS m/z287[M+H]⁺。¹H NMR(400MHz,DMSO-d6)δ7.98(2H,s)。

Intermediate 3

3-amino-6-bromo-5-trifluoromethylpyridin-2-formic acid

Intermediate 3A:2-bromo-3-nitro-5-trifluoromethylpyridin

3-nitro-5-(trifluoromethyl) pyridine-2-alcohol (31.00g, 149mmol) is dissolved in acetonitrile (250mL), obtains Dark brown solution.Add phosphorus oxybromide (V) (85g, 298mmol), and mixture is heated under reflux 4 hours, then exist RT is stirred overnight.By pouring into reactant mixture quencher in the water (600mL) containing sodium bicarbonate (110g) being stirred vigorously. Dark-brown mixture DCM (3 × 200mL) is extracted, and organic phase washed with water (200mL) and saline (100mL) is washed, It is dried (MgSO₄) and be concentrated in vacuo, obtain title product, for brown oil.¹H-NMR:[400MHz, CDCl₃,δ_H8.87 (1H, d, J=1.4Hz, ArH), 8.39 (1H, d, J=1.9Hz, ArH).

Intermediate 3B:3-nitro-5-trifluoromethylpyridin-2-nitrile

Bromo-for 2-3-nitro-5-trifluoromethylpyridin (10.00g, 36.87mmol) is under agitation dissolved in toluene (250mL), in, yellow solution is obtained.Add tetrabutyl ammonium bromide (11.90g, 36.9mmol), be subsequently added Cupricin. (I) (9.92g, 111mmol), and mixture is heated under reflux 9 hours.After being cooled to RT, by reactant mixture at water (750mL) and distribute between EtOAc (750mL).Merge organic fraction, wash with water (2 × 250mL) and saline (100mL), dry Dry (MgSO₄) and be concentrated in vacuo, obtain title product.¹H-NMR:[400MHz, DMSO-d₆δ_H9.55(1H,m,ArH),9.24 (1H,m,ArH)。

Intermediate 3C:3-amino-5-trifluoromethylpyridin-2-methyl formate

3-nitro-5-trifluoromethylpyridin-2-nitrile (6.5g, 29.9mmol) is dissolved in EtOAc (150mL), obtains yellowish Color solution.Add 10% palladium charcoal (3.19g, 2.99mmol), and reactant mixture is stirred in a hydrogen atmosphere 18 hours.Cross Filter reactant mixture and be concentrated in vacuo.Thick residue is dissolved in dense HCl (45mL), and is heated to backflow and reaches 24 hours.Will Reactant mixture is cooled to RT, and is concentrated in vacuo.Solid is dissolved in MeOH (200mL), and adds sulphuric acid (8mL).To produce Raw solution heats 84 hours under reflux.Reaction is cooled to RT, by adding 10%NaHCO_3(aq)(600mL) neutralize.Will Product is extracted in DCM (3 × 200mL), and by the organic phase washed with water (200mL) merged, saline (50mL) washing, is dried (MgSO₄) and be concentrated in vacuo.The solid of generation is passed through Silica gel chromatography: gradient: isohexane (500mL), 10%EtOAc in isohexane (1000mL), the 20%EtOAc in isohexane (1500mL), obtain title compound, for light Yellow solid.¹ H-NMR: [400MHz, DMSO-d₆,δ_H8.13 (1H, d, J=1.7Hz, ArH), 7.60 (1H, d, J=1.3Hz, ArH),7.01(2H,br,NH₂),3.85(3H,s,ArOCH₃), m/z221.1 [M+H]⁺。

Intermediate 3D:3-amino-6-bromo-5-trifluoromethylpyridin-2-methyl formate

3-amino-5-trifluoromethylpyridin-2-methyl formate (9.49g, 43.16mmol) is dissolved in water (300mL). Add sulphuric acid (4.60mL, 86mmol), go through subsequently 30 minutes dropping bromine (2.222mL, 43.1mmol) acetic acid (29.6mL, 517mmol) solution.Reactant mixture is stirred 18 hours at RT.Add 100mL water, add subsequently the bromine of 0.25 equivalent/ AcOH mixture (550 μ L bromines, in 7.4mL AcOH), and reactant mixture is stirred for 90 minutes at RT.Reaction is mixed Compound 500mL water dilutes, and adds solid NaHCO₃(～85g) neutralizes.Suspension DCM (3 × 300mL) is extracted, And the saturated NaHCO of organic facies that will merge_3(aq)(250mL), water (250mL) and saline (100mL) washing, be dried (MgSO₄) and be concentrated in vacuo.MeOH (～300mL) recrystallization seethed with excitement by thick material, obtains title product, for greenish orange Color solid.m/z301.0(95)[M+H]⁺。¹H-NMR:[400MHz, DMSO-d₆δ_H7.77(1H,s,ArH),7.17(2H,s, NH₂),3.86(3H,s,ArCO₂CH₃)。

Intermediate 3:3-amino-6-bromo-5-trifluoromethylpyridin-2-formic acid

3-amino-6-bromo-5-trifluoromethylpyridin-2-methyl formate (1.40g, 4.68mmol) is suspended in MeOH (15mL) in；Add sodium hydroxide (2.0M aqueous solution) (14.04mL, 28.1mmol), and suspension is stirred overnight at RT. Reactant mixture is concentrated in vacuo, and the residue of generation is dissolved in water (100mL), then by adding 5.0M HCl (aq) acidifying.Product is extracted in ethyl acetate (2 × 75mL), and by organic extract use water (50mL) merged, salt Water (25mL) washs, and is dried (MgSO₄) and be concentrated in vacuo, obtain title product, for yellow solid.¹H-NMR:[400MHz, DMSO-d₆,δ_H13.24(1H,br s,CO₂H),7.74(1H,s,ArH),7.1792H,br s ArNH₂)。m/z285.1, 287.1[M+H]⁺。

Intermediate 4

2-amino-5-iodo-4-(Trifluoromethyl)benzoic acid.

Intermediate 4A:2-amino-4-Trifluoromethyl-benzoic acid methyl ester

MeOH (750mL) solution of 2-amino-4-(trifluoromethyl) benzoic acid (75g, 366mmol) is gone through～15 minutes (exothermic temperature 34 DEG C) dropping sulphuric acid (48.7mL) processes, and maintains temperature at 20 DEG C.The mixture of generation is heated to 65 DEG C reach 24 hours.After being cooled to RT, mixture is under agitation diluted with water (750mL).The precipitate produced is filtered, and will be solid Body makes serosity in water (500mL).Adding saturated sodium bicarbonate (1L), pH is adjusted to pH9, and by mixture with ether (3 × 330mL) extraction.The organic extract saline (700mL) merged is washed, is dried (MgSO₄) and be concentrated in vacuo, marked Topic product.MS m/z220[M+H]⁺。

Intermediate 4B:2-amino-5-iodo-4-Trifluoromethyl-benzoic acid methyl ester

2-amino-4-Trifluoromethyl-benzoic acid methyl ester (48g, 219mmol) is dissolved in MeOH (750mL).Add sulphuric acid Silver (68.3g, 219mmol) and iodine (55.6g, 219mmol), and with MeOH (50mL) drip washing.Reactant mixture is put at RT Put 1 hour.Reactant mixture is filtered under vacuo, and filter vacuum is concentrated.Dissolve the residue in MeOH (100mL) and In water (10mL), and by mixture 80 DEG C of heating.Thick solution heat filtering is removed residue inorganic matter, and by the filter of temperature Liquid is poured slowly in water (750mL) with vigorous stirring.Mixture is placed 10 minutes and form granule, and filter, marked Topic compound, for orange solids.MS m/z345[M+H]⁺。

Intermediate 4:2-amino-5-iodo-4-(Trifluoromethyl)benzoic acid.

2-amino-5-iodo-4-Trifluoromethyl-benzoic acid methyl ester (5.0g, 14.49mmol) is dissolved in MeOH (100mL), Obtain orange solution.Add 2M NaOH (aq) (50.7mL, 101mmol), and orange solution is stirred overnight at RT.Will be anti- Should under reduced pressure remove MeOH, and add dense HCl in the solution of stirring_(aq)It is acidified remaining aqueous solution, until forming thickness Precipitate.Cross filter solid, wash with water (2 × 10mL), be dried 48 hours in vacuum and 40 DEG C subsequently, obtain title compound Thing, for yellow/brown solid.¹H NMR(400MHz,DMSO-d6)δ7.98(2H,s)。

Intermediate 5

3-amino-6-chloro-5-trifluoromethylpyridin-2-formic acid

By molten for 3-amino-6-bromo-5-trifluoromethylpyridin-2-methyl formate (intermediate 3D) (100mg, 0.334mmol) In 5M HCl (2.5mL), and 150 DEG C, 5.5 bar microwaves heat 1 hour.Reactant mixture is passed through reverse-phase chromatography Purification, with water/MeCN eluting, obtains title compound.MS m/z241[M+H]⁺。¹H NMR(400MHz,DMSO-d6)δ13.33 (1H, br hump), 7.79 (1H, s), 7.19 (2H, br s).

Intermediate 6

3-amino-5-trifluoromethyl-pyrazine-2-formic acid

To the 3-amino-5-trifluoromethyl-pyrazine-2-Ethyl formate (intermediate 1) (0.5g, 2.126mmol) of stirring Drying ethanol (5mL) solution adds 2M NaOH (5.32mL, 10.63mmol).Yellow solution is stirred at RT.After 5 minutes, Reactant mixture is poured in water (15mL), and by adding 1M HCl, pH is adjusted to pH6.The yellow mercury oxide formed is existed Filtered under vacuum, and it is dried to obtain title compound.¹H NMR(400MHz,DMSO-d6)δ13.65(1H,s),8.3(1H, s),7.86(2H,br s)。

Intermediate 7

3-amino-6-chloro-5-trifluoromethyl-pyrazine-2-formic acid

Intermediate 7A: 3-amino-6-chloro-5-trifluoromethyl-pyrazine-2-Ethyl formate

Will be at 4M HCl (twoAlkane (2mL)) in 3-amino-6-bromo-5-trifluoromethyl-pyrazine-2-formic acid (middle Body 2) (200mg, 0.637mmol, 1 equivalent) heat 30 minutes with microwave radiation at 110 DEG C, then 1.5 little 100 DEG C of heating Time.Reactant mixture is poured in water (30mL), and use saturated NaHCO₃Neutralize.Product EtOAc (2 × 20mL) is extracted Take, merge Organic substance and wash with saline (20mL), being dried (MgSO₄) and be concentrated in vacuo, obtain yellow oil.Will be thick Material is dissolved in MeOH (3mL), and grinds with water (15mL), obtains title compound, for faint yellow solid.MS m/z270 [M+H]⁺。¹H NMR (400MHz, DMSO-d6) δ 7.96 (2H, wide hump), 4.4 (2H, q), 1.34 (3H, t).

Intermediate 7:3-amino-6-chloro-5-trifluoromethyl-pyrazine-2-formic acid

To stirring 3-amino-6-chloro-5-trifluoromethyl-pyrazine-2-Ethyl formate (intermediate 7A) (165mg, Drying ethanol (10mL) solution 0.612mmol) adds 2M NaOH (0.306mL, 0.612mmol).Stir 10 minutes at RT After, reactant mixture is poured in water (30mL), and add 1M HCl pH is adjusted to 6.By product with EtOAc (2 × 30mL) Extraction.Merge Organic substance and wash with saline (20mL), being dried (MgSO₄) and be concentrated in vacuo.By thick material by preparation Type LCMS purification, uses 0.1%TFA/MeCN/ aqueous systems, obtains title compound.MS m/z244[M+H]⁺。

Intermediate 8

3-(piperidin-1-yl methyl)-1H-pyrazoles-5-amine

NaH (0.234g, 5.84mmol) in dry toluene (5mL) is heated to 80 DEG C.The reactant mixture of thermotropism Middle dropping acetonitrile (0.229mL, 4.38mmol), 1-ethyl piperidine (0.5g, 2.92mmol) and the mixing of toluene (3mL) Thing.Mixture is stirred 1.5 hours at 80 DEG C, after being cooled to RT, by adding water (6mL) quencher.Add hydrazine hydrate (0.142mL, 2.92mmol), and reactant mixture is again heated to 80 DEG C reaches 3 hours.Reactant mixture is used 2N HCl (aq.) it is acidified to pH2, and loads SCX-2 post, use MeOH eluting, the 7M NH being subsequently used in MeOH₃Eluting.It is concentrated in vacuo Methanol ammonia fraction, obtains grease.Two it are added in yellow oil4N HCl in alkane (15mL), and filter product Raw suspension, and with iso-hexane, obtain title compound.

MS m/z181.1[M+H]⁺。¹H NMR(400MHz,DMSO-d6)δ10.95(1H,br),6.28(1H,br),4.24 (2H,s),3.30(2H,br d),2.83(2H,br),1.78(4H,m),1.68(2H,br d),1.37(2H,br)。

Following is the further embodiment of the present invention:

Embodiment 1: compound of formula I

Or its officinal salt or solvate, wherein:

A is N or CR⁴；

B is N or CR⁵；

D is N or CR⁶；

E is N or CR⁷, condition be D and E be not all N；

R¹Selected from H；The C optionally replaced by one or more halogen atoms₁-C₈Alkyl；C₂-C₈Alkenyl；C₂-C₈Alkynyl； C₃-C₁₀Cycloalkyl；C₅-C₁₀Cycloalkenyl group；-C₁-C₄Alkyl-C₃-C₈Cycloalkyl；Optionally replaced by one or more halogen atoms C₁-C₈Alkoxyl；OH；Halogen；SO₂NR⁸R⁹；SO₂R¹⁰；The S-C optionally replaced by one or more halogen atoms₁-C₈Alkyl；S- C₆-C₁₄Aryl；-(C₀-C₄Alkyl)-C₆-C₁₄Aryl；-(C₀-C₄Alkyl)-3 to 14 yuan of heterocyclic radicals, wherein heterocyclic radical comprises at least One hetero atom selected from N, O and S；CN；NR¹¹R¹²；CONR¹³R¹⁴；NR¹³SO2R¹⁵；NR¹³C(O)R¹⁵And CO₂R¹⁵, wherein cycloalkanes Base, cycloalkenyl group, aryl and heterocyclic radical are the most optionally replaced by one or more Z substituent groups；

R²Selected from the C optionally replaced by one or more halogen atoms₁-C₈Alkyl；C₂-C₈Alkenyl；C₂-C₈Alkynyl；C₃- C₁₀Cycloalkyl；C₅-C₁₀Cycloalkenyl group；-C₁-C₄Alkyl-C₃-C₈Cycloalkyl；The C optionally replaced by one or more halogen atoms₁-C₈ Alkoxyl；OH；Cl；Br；I；SO₂NR⁸R⁹；SO₂R¹⁰；The S-C optionally replaced by one or more halogen atoms₁-C₈Alkyl；Optionally The S-C replaced by one or more Z substituent groups₆-C₁₄Aryl；-(C₀-C₄Alkyl)-C₆-C₁₄Aryl；-(C₀-C₄Alkyl)-3 to 14 Unit's heterocyclic radical, wherein heterocyclic radical comprises at least one hetero atom selected from N, O and S；CN；CONR¹³R¹⁴；NR¹³SO2R¹⁵；NR¹³C (O)R¹⁵And CO₂R¹⁵, wherein cycloalkyl, cycloalkenyl group, aryl and heterocyclic radical are the most optionally replaced by one or more Z substituent groups；

R⁶And R⁷It is each independently selected from following formula group:

-(CH₂)_n-X-(CH₂)_m-NR¹⁷R¹⁸；Or

R⁶And R⁷Carbon atom in connection forms 5 to 8 yuan of carbocyclic ring systems or together containing one or more selected from N, O With heteroatomic 5 to 8 yuan of heterocycle systems of S, wherein ring system is optionally replaced by one or more Z substituent groups；

X be do not exist, O, S, CO, SO, SO₂Or CH₂；

N and m is each independently selected from 0,1,2 and 3；

R⁹、R¹⁰、R¹²、R¹⁴、R¹⁵、R¹⁶And R¹⁸It is each independently selected from H；Optionally replaced by one or more halogen atoms C₁-C₈Alkyl；C₂-C₈Alkenyl；C₂-C₈Alkynyl；C₃-C₁₀Cycloalkyl；C₅-C₁₀Cycloalkenyl group；-C₁-C₄Alkyl-C₃-C₈Cycloalkyl；- (C₀-C₄Alkyl)-C₆-C₁₄Aryl；With-(C₀-C₄Alkyl)-3 to 14 yuan of heterocyclic radicals, wherein heterocyclic radical comprises at least one and is selected from The hetero atom of N, O and S, wherein cycloalkyl, cycloalkenyl group, aryl and heterocyclic radical are the most optionally taken by one or more Z substituent groups Generation；Or

Z is independently selected from OH；Aryl；O-aryl；Benzyl；O-benzyl；Optionally by one or more OH groups or NH₂Group Substituted C₁-C₆Alkyl；The C optionally replaced by one or more halogen atoms₁-C₆Alkyl；Optionally by one or more OH groups Or C₁-C₄The substituted C of alkoxyl₁-C₆Alkoxyl；NR^18’(SO₂)R²¹；(SO₂)NR¹⁹R²¹；(SO₂)R²¹；NR^18’C(O)R²¹；C(O) NR¹⁹R²¹；NR¹⁹C(O)NR¹⁹R²¹；NR^18’C(O)OR¹⁹；NR¹⁹R²¹；C(O)OR¹⁹；C(O)R¹⁹；SR¹⁹；OR¹⁹；Oxo；CN；NO₂； Halogen；With-(C₀-C₄Alkyl)-3 to 14 yuan of heterocyclic radicals, wherein heterocyclic radical comprises at least one hetero atom selected from N, O and S, appoints Select by one or more selected from halogen, oxo, C₁-C₆Alkyl and C (O) C₁-C₆Alkyl replaces；

R^18’And R²⁰It is each independently selected from H and C₁-C₆Alkyl；

R¹⁹And R²¹It is each independently selected from H；C₁-C₈Alkyl；C₃-C₈Cycloalkyl；C₁-C₄Alkoxy-C₁-C₄Alkyl；Optionally By one or more selected from C₁-C₆Alkyl, C₁-C₆Alkoxyl and the (C of halogen substiuted₀-C₄Alkyl)-aryl；(C₀-C₄Alkyl)-3- To 14-unit heterocyclic radical, heterocyclic radical includes one or more hetero atom selected from N, O and S, optionally by one or more selected from halogen Element, oxo, C₁-C₆Alkyl and C (O) C₁-C₆The group of alkyl replaces；Optionally by one or more selected from C₁-C₆Alkyl, C₁-C₆Alkane Substituted (the C of group of epoxide and halogen₀-C₄Alkyl)-O-aryl；(C₀-C₄Alkyl)-O-3-to 14-unit heterocyclic radical, heterocycle Base comprises one or more hetero atom selected from N, O and S, optionally by one or more selected from halogen, C₁-C₆Alkyl and C (O) C₁- C₆The group of alkyl replaces；Wherein alkyl is optionally by one or more halogen atoms, C₁-C₄Alkoxyl, C (O) NH₂、C(O)NHC₁- C₆Alkyl or C (O) N (C₁-C₆Alkyl)₂Replace；Or

R¹⁹And R²¹Forming 5-to 10-unit heterocyclic radical together with the nitrogen-atoms being connected with them, heterocyclic radical comprises one or more Selected from the other hetero atom of N, O and S, heterocyclic radical is optionally replaced selected from following substituent group by one or more: OH；Halogen； Aryl；Comprise one or more heteroatomic 5-to 10-unit heterocyclic radical selected from N, O and S；S(O)₂-aryl；S(O)₂-C₁-C₆ Alkyl；The C optionally replaced by one or more halogen atoms₁-C₆Alkyl；Optionally by one or more OH groups or C₁-C₄Alcoxyl The substituted C of base₁-C₆Alkoxyl；With C (O) OC₁-C₆Alkyl, wherein aryl and heterocyclic substituent self are optionally by C₁-C₆Alkyl, C₁-C₆Haloalkyl or C₁-C₆Alkoxyl replaces；

For treating disease or the obstacle of CFTR mediation.

Embodiment 2: according to the compound of embodiment 1, wherein A is N.

Embodiment 3: according to embodiment 1 or the compound of embodiment 2, wherein R¹Selected from H；Optionally by one or many The substituted C of individual halogen atom₁-C₈Alkyl；Halogen；C₆-C₁₄Aryl；-(C₀-C₄Alkyl)-3 to 14 yuan of heterocyclic radical, wherein heterocyclic radicals Comprise at least one hetero atom selected from N, O and S；And NR¹¹R¹², wherein aryl and heterocyclic radical are the most optionally by one or more Z Substituent group replaces.

Embodiment 4: according to the compound of any one, wherein R in the embodiment above²It is the most one or more The substituted C of halogen atom₁-C₈Alkyl；C₃-C₁₀Cycloalkyl；Or-C₁-C₄Alkyl-C₃-C₈Cycloalkyl.

Embodiment 5: according to the compound of embodiment 4, wherein R²It is CF₃。

Embodiment 6: according to the compound of any one, wherein R in the embodiment above³It is H or methyl.

Embodiment 7: according to the compound of any one, wherein R in the embodiment above^aIt is H.

Embodiment 8: according to the compound of any one in the embodiment above, wherein E is CR⁷。

Embodiment 9: according to the compound of embodiment 1, wherein

A is N

R¹Selected from H；Halogen；C₆-C₁₄Aryl；5 or 6-unit's heterocyclic radicals, wherein heterocyclic radical comprises at least one selected from N, O and S Hetero atom；And NR¹¹R¹², wherein aryl and heterocyclic radical are the most optionally replaced by one or more Z substituent groups；

R²It is CF₃；

R^aIt is H；

R³It is H or Me；And

E is CR⁷。

Embodiment 10: compound of formula I, it is selected from following compounds:

Embodiment 11: compound of formula I, it is selected from following compounds:

3-amino-5-trifluoromethyl-pyrazine-2-formic acid (2H-pyrazole-3-yl)-amide；

3-amino-6-[4-(5-methyl-[1,3,4]Diazole-2-base)-phenyl]-5-trifluoromethylpyridin-2-formic acid (2H-pyrazole-3-yl)-amide；

Or its officinal salt or solvate.

Embodiment 12: compound of formula I

Or its officinal salt or solvate, wherein R¹、R²、R³、R^a, A, B, D and E such as embodiment 1 to 11 any one Individual definition, condition be A or B or both be N.

Embodiment 13: comprise the compound according to embodiment 12 and one or more pharmaceutically acceptable loads of therapeutically effective amount The pharmaceutical composition of body.

Embodiment 14: comprise compound and the one of any one definition of the embodiment 1 to 12 of therapeutically effective amount Or the combination of multiple therapeutic activity activating agent altogether.

Embodiment 15: according to the combination of embodiment 14, wherein said common activating agent blocks selected from penetrating agent, ENaC Agent, antiinflammatory, bronchodilator, hydryllin, anti-tussive agents, antibiotic and deoxyribonuclease medicine, wherein the first Can be in identical or different pharmaceutical composition with the second activating agent.

Embodiment 16: according to the compound of embodiment 12, it is used as medicine.

Embodiment 17: according to the compound of any one of embodiment 1 to 12, it is used for treating obstacle or disease, Described obstacle or disease selected from inflammatory or obstructive airway diseases or mucosal hydration.

Embodiment 18: according to the compound of any one of embodiment 1 to 12, it is used for treating obstacle or disease, Described obstacle or disease are selected from cystic fibrosis, primary ciliary dyskinesia, chronic bronchitis, Chronic obstructive pulmonary disease (IBS, IBD, opioid lure for disease, asthma, respiratory tract infection, pulmonary carcinoma, xerostomia and keratoconjunctivitis sicca or constipation Lead).

Embodiment 19: the compound of any one definition of embodiment 1 to 12 is used for treating CFTR mediation in preparation Disease or obstacle medicine in purposes.

Embodiment 20: the compound of any one definition of embodiment 1 to 12 is used for treating obstacle or disease in preparation The sick purposes in medicine, described obstacle or disease selected from inflammatory or obstructive airway diseases or mucosal hydration.

Embodiment 21: the compound of any one definition of embodiment 1 to 12 is used for treating obstacle or disease in preparation The sick purposes in medicine, described obstacle or disease are selected from cystic fibrosis, primary ciliary dyskinesia, chronic bronchial Inflammation, chronic obstructive pulmonary disease, asthma, respiratory tract infection, pulmonary carcinoma, xerostomia and keratoconjunctivitis sicca or constipation (induction of IBS, IBD, opioid).

Embodiment 22: treating disease or the method for obstacle of CFTR mediation in individuality, wherein the method includes to individuality The compound of any one definition of the embodiment 1 to 12 of administering therapeutic effective dose.

Embodiment 23: the method for CFTR activity in regulation individuality, wherein the method includes to individual administering therapeutic effective The compound of any one definition of the embodiment 1 to 12 of amount.

Embodiment 24: according to the method for any one of embodiment 22 to 23, wherein obstacle or disease are selected from capsule Fibrosis, primary ciliary dyskinesia, chronic bronchitis, chronic obstructive pulmonary disease, asthma, respiratory tract infection, pulmonary carcinoma, Xerostomia and keratoconjunctivitis sicca or constipation (induction of IBS, IBD, opioid).

Embodiment 25: for treating obstacle or the pharmaceutical composition of disease of CFTR mediation, it comprises to become as activity The compound of any one according to embodiment 1 to 12 divided.

Embodiment 26: the pharmaceutical composition of embodiment 26, wherein obstacle or disease are selected from cystic fibrosis, constitutional Ciliary dyskinesia, chronic bronchitis, chronic obstructive pulmonary disease, asthma, respiratory tract infection, pulmonary carcinoma, xerostomia and dry Dry property keratoconjunctivitis or constipation (induction of IBS, IBD, opioid).

Claims

1. compound of formula I

Or its officinal salt, wherein:

A is N；

B is CR⁵；

D is CR⁶；

E is CR⁷；

R¹Selected from halogen；C₆-C₁₄Aryl；With 5 or 6-unit's heterocyclic radicals, wherein heterocyclic radical comprises at least one miscellaneous selected from N, O and S Atom；Wherein aryl and heterocyclic radical are the most optionally replaced by one or more Z substituent groups；

R²It is CF₃；

R^aIt is H；

R³It is H；

R⁵It is H；

R⁶And R⁷It is each independently selected from H；The C optionally replaced by one or more halogen atoms₁-C₈Alkyl；

Z is independently selected from C₁-C₆Alkyl；C₁-C₆Alkoxyl；Halogen；With-(C₀-C₄Alkyl)-3 to 14 yuan of heterocyclic radical, wherein heterocycles Base comprises at least one hetero atom selected from N, O and S, optionally by one or more selected from halogen, oxo, C₁-C₆Alkyl and C (O) C₁-C₆The group of alkyl replaces.

2. compound of formula I, it is selected from following compounds:

3-amino-6-(4-chloro-2-methyl-phenyl)-5-trifluoromethylpyridin-2-formic acid (2H-pyrazole-3-yl)-amide；

3-amino-6-[3-(2-oxo-pyrroli-1-ylmethyl)-phenyl]-5-trifluoromethylpyridin-2-formic acid (2H-pyrrole Azoles-3-base)-amide；With

Or its officinal salt.

3. comprise the compound of the claim 1 or 2 of therapeutically effective amount and the pharmaceutical composition of one or more pharmaceutically suitable carrier.

4. comprise compound and the combination of one or more therapeutic activities activating agent altogether of the claim 1 or 2 of therapeutically effective amount Product.

5. the combination product of claim 4, wherein said common activating agent is selected from penetrating agent, ENaC blocker, antiinflammatory, a gas Enlargement of pipe agent, hydryllin, anti-tussive agents, antibiotic and deoxyribonuclease medicine.

6. the compound of claim 1 or 2, it is used as medicine.

7. the compound of claim 1 or 2, it is used for treating obstacle or disease, described obstacle or disease selected from inflammatory or resistance Plug property airway disorders or the disease be benefited from mucosal hydration.

8. the compound of claim 1 or 2 in preparation for treating the purposes in the medicine of obstacle or disease, described obstacle or Disease selected from inflammatory or obstructive airway diseases or the disease be benefited from mucosal hydration.