JP2010539204A - Small molecule bradykinin B1 receptor antagonist - Google Patents

Abstract

ブラジキニンＢ１受容体（Ｂ１Ｒ）アンタゴニストである式（Ｉ）の化合物を開示する。これらの化合物は、炎症及び痛みと関連する疾患の処置又は不利な症状の緩和に有用である。本発明は、新規な化合物及び許容され得るその誘導体、製薬学的組成物ならびに炎症及び痛みを含む疾患の予防及び処置のための方法を包含する。[Chemical 1]

Disclosed are compounds of formula (I) that are bradykinin B1 receptor (B1R) antagonists. These compounds are useful for the treatment of diseases associated with inflammation and pain or for alleviating adverse symptoms. The present invention includes novel compounds and acceptable derivatives thereof, pharmaceutical compositions and methods for the prevention and treatment of diseases including inflammation and pain.

Description

  The present invention is responsive to the field of biologically active pharmaceutical compounds and specifically to selective antagonists to the bradykinin B1 receptor (B1R) and bradykinin B1 receptors such as inflammation and pain-related disorders. It relates to their use for the treatment of certain conditions and diseases.

Kinin's biological effects are mediated by two major G-protein coupled receptors, B1R and B2R. Although B2R is constitutively expressed in a variety of cells under physiological conditions, B1R is found in several cell types including endothelial cells, smooth muscle cells, blood cells and neurons, such as tissue damage or inflammation. It is induced under pathophysiological conditions (Non-patent document 1; Non-patent document 2). This makes B1R a particularly attractive drug target. Activation of B1R is associated with edema, pain and blood-derived leukocyte trafficking.
Produces a range of pro-inflammatory effects that include the promotion of trafficking (3).

  Bradykinin (BK) and kallidin (KD) are peptide kinins that act on B2R and mediate the acute physiological effects of kinin on the cardiovascular, renal, nervous and immune systems. BK and kallidin are metabolized by carboxypeptidases N and M, which remove the carboxy-terminal arginine residue to produce des-Arg-9-BK (DABK) or des-Arg-10-kallidine (DAKD). While DAKD is the only known natural ligand for human B1R, des-Arg-9-BK activates B1R in rodents. DAKD that specifically acts on human B1R appears to be an important mediator of inflammation and pain in humans (Non-Patent Document 4).

  Numerous peptides and non-peptide antagonists of B1R have been described in the prior art as novel therapeutics for the treatment of pain and inflammation (Non-Patent Document 5). For example, Patent Literature 1, Patent Literature 2, Patent Literature 3, Patent Literature 4, and Patent Literature 5 disclose biphenyl compounds that are B1R antagonists, and Patent Literature 6 discloses a sulfonamide compound.

  In view of the overshooting and thus the critical conditions associated with both acute and chronic pathological inflammation and pain, there is a need for new compounds that are effective in the selective blockade of B1 receptor activation.

International Publication No. 030666577 Pamphlet International Publication No. 030565789 Pamphlet Pamphlet of International Publication No. 0516886 International Publication No. 04019868 Pamphlet US Patent No. 20060122236 International Publication No. 9725315 Pamphlet

By Regoli and Barbe, Pharmacol. Rev. 32, 1980, 1-46 Marceau et al. Author, Pharmacol. Rev. 50, 1998, 357-386 Calixto et al. Author, Br. J. et al. Pharmacol. 143, 2004, 803-818 Leeb-Lundberg et al. Author, Pharmacol. Rev. 57, 2005, 27-77 Chen et al. Author, Expert Opin. Ther. Targets 11, 2007, 21-35

  Thus, the problem underlying the present invention is to provide highly selective B1R antagonists that have improved properties, preferably over prior art B1R antagonists.

  In the first embodiment of the first aspect, the problem underlying the present invention is formula (I):

Or a pharmacologically acceptable salt, solvate or hydrate thereof, wherein
A is: i) an optionally substituted 5- or 6-membered cycloalkyl;
ii) an optionally substituted 5- or 6-membered heterocycloalkyl;
iii) optionally substituted 6-aryl; or iv) optionally substituted 5- or 6-membered heteroaryl;
T is a hydrogen atom or bonded to B i) forming an optionally substituted heterocycloalkyl; or ii) forming an optionally substituted heteroaryl;
B is an optionally substituted alkyl, an optionally substituted alkenyl, an optionally substituted alkynyl, an optionally substituted heteroalkyl, optionally substituted Optionally substituted cycloalkyl, optionally substituted heterocycloalkyl, optionally substituted alkylcycloalkyl, optionally substituted heteroalkylcycloalkyl, if An aryl which can be optionally substituted, an optionally substituted heteroaryl, an optionally substituted aralkyl, an optionally substituted heteroaralkyl or

And
W is N, alkyl, heteroalkyl, alkenyl, cycloalkyl, alkylcycloalkyl, heteroalkylcycloalkyl, aryl, heteroaryl, aralkyl or heteroaralkyl;
R ⁴ , if present, is a hydrogen atom, an optionally substituted alkyl, an optionally substituted heteroalkyl, an optionally substituted alkylcycloalkyl, optionally A heteroalkylcycloalkyl that may be substituted, an aryl that may be optionally substituted, an aralkyl that may be optionally substituted, a heteroaralkyl that may be optionally substituted, or Bonded to R ⁵ and together with W
i) an optionally substituted cycloalkyl;
ii) an optionally substituted heterocycloalkyl;
iii) an optionally substituted aryl; or iv) forming an optionally substituted heteroaryl;
R ⁵ , if present, is a hydrogen atom, an optionally substituted alkyl, an optionally substituted heteroalkyl, an optionally substituted heteroalkylcycloalkyl, An aryl that can be substituted by, an aralkyl that can be optionally substituted, a heteroaralkyl that can be optionally substituted, or bonded to R ⁴ and taken together with W;
i) an optionally substituted cycloalkyl;
ii) an optionally substituted heterocycloalkyl;
iii) an optionally substituted aryl; or iv) forming an optionally substituted heteroaryl;
E, if present, is heteroalkyl or heteroaralkyl;
G, if present, is alkyl, heteroalkyl, cycloalkyl, heterocycloalkyl, alkylcycloalkyl, heterocycloalkylalkyl, heteroaryl or heteroaralkyl;
K is

And
Here, R ¹ and R ² are each independently selected from C ₂ -C ₆ alkyl, C ₂ -C ₆ alkenyl, C ₂ -C ₆ alkynyl, C ₂ -C ₆ heteroalkyl, or together with N To form an optionally substituted heterocycloalkyl, an optionally substituted heteroaryl or an optionally substituted heteroaralkyl; and R ³ is a hydrogen atom. , C ₁ -or C ₂ alkyl, cyano or a halogen atom.

  In the second aspect of the first aspect, which is also one aspect of the first aspect of the first aspect, A is

Where X ¹ , X ² and X ³ are each independently selected from N, O, S, NR ^x , CR ^x or CR ^x R ^{x ′} , where R ^x and R ^{x ′} are each independently Hydrogen atom, halogen atom, ═O, hydroxy, cyano, amino, nitro, mercapto, alkyl, alkenyl, alkynyl, heteroalkyl, cycloalkyl, heterocycloalkyl, alkylcycloalkyl, heteroalkylcycloalkyl, aryl, heteroaryl , Aralkyl or heteroaralkyl.

  In the third embodiment of the first aspect, which is also one embodiment of the first or second embodiment of the first aspect, A is

Chosen from
Here, R ^x , R ^x1 , R ^x2 , R ^x3 , R ^{x ′} , R ^{x1 ′} , R ^{x2 ′} and R ^{x3 ′} are each independently a hydrogen atom, a halogen atom, ═O, hydroxy, cyano, amino, nitro , Mercapto, alkyl, alkenyl, alkynyl, heteroalkyl, cycloalkyl, heterocycloalkyl, alkylcycloalkyl, heteroalkylcycloalkyl, aryl, heteroaryl, aralkyl or heteroaralkyl; and X is NH, O or S is there.

  In the fourth embodiment of the first aspect, which is also one embodiment of the first embodiment of the first aspect, A is

And
Here, Y ¹ , Y ² , Y ³ and Y ⁴ are each independently selected from N, O, S, NR ^y , CR ^y or CR ^y R ^{y ′} , where R ^y and R ^{y ′} are Each independently a hydrogen atom, a halogen atom, = O, hydroxy, cyano, amino, nitro, mercapto, alkyl, alkenyl, alkynyl, heteroalkyl, cycloalkyl, heterocycloalkyl, alkylcycloalkyl, heteroalkylcycloalkyl, aryl, Selected from heteroaryl, aralkyl or heteroaralkyl.

  In the fifth aspect of the first aspect, which is also one aspect of the first aspect or the fourth aspect of the first aspect, A is

Chosen from
Here, R ^y , R ^y1 , R ^y2 , R ^y3 , R ^y4 , R ^{y ′} , R ^{y1 ′} , R ^{y2 ′} , R ^{y3 ′} and R ^{y4 ′} are each independently a hydrogen atom, a halogen atom, ═O, It is selected from hydroxy, cyano, amino, nitro, mercapto, alkyl, alkenyl, alkynyl, heteroalkyl, cycloalkyl, heterocycloalkyl, alkylcycloalkyl, heteroalkylcycloalkyl, aryl, heteroaryl, aralkyl or heteroaralkyl.

  In a sixth aspect of the first aspect, which is also an aspect of the first, fourth or fifth aspect of the first aspect, A is

And
Here, R ^y1 , R ^y2 , R ^y3 and R ^y4 are each independently a hydrogen atom, a halogen atom, ═O, hydroxy, cyano, amino, nitro, mercapto, alkyl, alkenyl, alkynyl, heteroalkyl, cycloalkyl, hetero It is selected from cycloalkyl, alkylcycloalkyl, heteroalkylcycloalkyl, aryl, heteroaryl, aralkyl or heteroaralkyl.

In the seventh aspect of the first aspect, which is also one aspect of the sixth aspect of the first aspect,
R ^y1 is a hydrogen atom.

In the eighth aspect of the first aspect, which is also one aspect of the sixth or seventh aspect of the first aspect,
R ^y2 is a hydrogen atom, a halogen atom or C ₁ -C ₆ alkyl; and R ^y4 is a hydrogen atom or a halogen atom.

In the ninth aspect of the first aspect, which is also one aspect of the sixth to eighth aspects of the first aspect, R ^y2 is a hydrogen atom.

In the tenth embodiment of the first aspect, which is also one of the sixth to ninth embodiments of the first aspect, R ^y3 is a hydrogen atom or a halogen atom.

In the eleventh embodiment of the first aspect, which is also one of the sixth to tenth aspects of the first aspect, R ^y4 is a hydrogen atom or a halogen atom.

  In the twelfth embodiment of the first aspect, which is also one of the first to eleventh aspects of the first aspect, T is a hydrogen atom.

  In the thirteenth aspect of the first aspect, which is also one aspect of the first to twelfth aspects of the first aspect, K is

Chosen from.

  In the fourteenth embodiment of the first aspect, which is also one of the first to thirteenth embodiments of the first aspect, K is

Chosen from.

  In the fifteenth aspect of the first aspect, which is also one of the first to fourteenth aspects of the first aspect, K is

It is.

  In the sixteenth embodiment of the first aspect, which is also one of the first to fourteenth embodiments of the first aspect, K is

It is.

  In a seventeenth embodiment of the first aspect which is also an embodiment of the first to sixteenth embodiments of the first aspect, B is heteroalkyl, heteroaryl or

And
W is N, alkyl, alkenyl, cycloalkyl, alkylcycloalkyl, heteroalkylcycloalkyl, aryl, heteroaryl, aralkyl or heteroaralkyl;
R ⁴ , if present, is a hydrogen atom, alkyl, heteroalkyl, alkylcycloalkyl, heteroalkylcycloalkyl, aryl, aralkyl, heteroaralkyl, or bonded to R ⁵ and together with N
i) an optionally substituted cycloalkyl;
ii) an optionally substituted heterocycloalkyl;
iii) optionally substituted aryl; or iv) optionally substituted heteroaryl; and R ⁵ , if present, is a hydrogen atom, optionally substituted An optionally substituted heteroalkyl, an optionally substituted heteroalkyl, an optionally substituted heteroalkylcycloalkyl, an optionally substituted aryl, optionally substituted. An aralkyl which can be optionally substituted heteroaralkyl, or bonded to R ⁴ and taken together with N;
i) an optionally substituted cycloalkyl;
ii) an optionally substituted heterocycloalkyl;
iii) an optionally substituted aryl; or iv) an optionally substituted heteroaryl.

In the eighteenth aspect of the first aspect, which is also one aspect of the first to seventeenth aspects of the first aspect,
B is ^-Y aB ^{-CO-L B} - or ^-Y aB ^{-CO-R cB} - a, where in ^{Y aB} is a _bond, C 1 -C _{6 alkylene,} C 2 -C ₆ alkenylene or _C 2 -C ₆ Alkynylene;
L ^B is a cycloalkyl, heterocycloalkyl, alkylcycloalkyl, heteroaryl alkyl cycloalkyl, optionally aryl which may be substituted, optionally heteroaryl may be substituted, aralkyl or heteroaralkyl; and R ^cB is an optionally substituted C ₁ -C ₈ alkyl, an optionally substituted C ₂ -C ₈ alkenyl or an optionally substituted C ₂ -C ₈ Alkynyl.

  In the nineteenth aspect of the first aspect, which is also one of the first to eighteenth aspects of the first aspect, B is

It is.

  In the twentieth aspect of the first aspect, which is also one aspect of the first to eighteenth aspects of the first aspect, B is

And
Where R ⁴ is methyl, ethyl or isopropyl.

  In the twenty-first aspect of the first aspect, which is also one aspect of the first to seventeenth aspects of the first aspect, B is

It is.

In a 22 embodiment of the first aspect which is also one aspect of the first to 21 embodiment of the first aspect, E is ^{-Y aE -CO-L E -,} - Y aE -CO-R cE - ^{, -Y aE -NR cE -CO-R} dE -, - Y aE -NR cE -CO-L E -, - Y aE -NR cE -CO-NR dE -L E - or ^-Y aE ^{-NR cE} - ^CS-NR dE -L ^E - a, where in ^{Y aE} the _bond, C 1 -C _{6 alkylene,} be C 2 -C ₆ alkenylene or _C 2 -C ₆ alkynylene;
R ^cE is a hydrogen atom, an optionally substituted C ₁ -C ₈ alkyl, an optionally substituted C ₂ -C ₈ alkenyl or an optionally substituted C _2- C ₈ alkynyl, provided that in —Y ^aE —CO—R ^cE —, R ^cE is not a hydrogen atom;
R ^dE is a hydrogen atom, an optionally substituted C ₁ -C ₈ alkyl, an optionally substituted C ₂ -C ₈ alkenyl or an optionally substituted C _2- be a C ₈ alkynyl; and L ^E is cycloalkyl, heterocycloalkyl, alkylcycloalkyl, heteroalkyl cycloalkyl, optionally aryl which may be substituted, optionally heteroaryl may be substituted, aralkyl Or heteroaralkyl.

In the twenty-third embodiment of the first aspect, which is also one of the first to twenty-first embodiments of the first aspect, G is alkyl, cycloalkyl, —Y ^aG —O—R ^cG , —Y ^aG —CO ^{-NR aG R bG, -Y aG -S} -R cG, -Y aG -SO-R cG, -Y aG -SO 2 -R cG, heteroaryl, alkyl cycloalkyl, or heterocycloalkyl,
Where Y ^aG is a bond, C ₁ -C ₆ alkylene, C ₂ -C ₆ alkenylene or C ₂ -C ₆ alkynylene;
R ^aG is a hydrogen atom, C ₁ -C ₆ alkyl, C ₂ -C ₆ alkenyl, C ₂ -C ₆ alkynyl, or bonded to R ^bG to form a 4- to 10-membered cycloalkyl or heterocycloalkyl. Forming;
R ^bG is a hydrogen atom, C ₁ -C ₆ alkyl, C ₂ -C ₆ alkenyl, C ₂ -C ₆ alkynyl, or is bonded to R ^aG to form a ^4- to 10-membered cycloalkyl or heterocycloalkyl. And R ^cG is an optionally substituted C ₁ -C ₈ alkyl, an optionally substituted C ₂ -C ₈ alkenyl or an optionally substituted C ₂ -C is ₈ alkynyl.

  In a twenty-fourth embodiment of the first aspect, which is also an embodiment of the first, fourth to sixth, twelfth, seventeenth or twenty-second embodiments of the first aspect, the compound has the formula (II):

Indicated by
Where
Y ¹ is N, CH or CF;
K is

Chosen from;
R ³ is a hydrogen atom or methyl;
R ^y2 is a hydrogen atom, a halogen atom or C ₁ -C ₆ alkyl;
R ^y3 is a hydrogen atom, a halogen atom, C ₁ -C ₆ alkyl or C ₁ -C ₆ heteroalkyl;
R ^y4 is a hydrogen atom or a halogen atom;
The ^{B -Y aB -CO-L B -} , - Y aB -CO-R cB - or

And
Where Y ^aB is a bond;
R ^cB is an optionally substituted C ₁ -C ₈ alkyl or an optionally substituted C ₂ -C ₈ alkenyl;
L ^B is cycloalkyl, heterocycloalkyl, optionally aryl which may be substituted, optionally heteroaryl or heteroaralkyl may be substituted;
W is alkyl or N;
R ⁴ , if present, is a hydrogen atom, alkyl, alkylcycloalkyl, heteroalkylcycloalkyl, aralkyl or heteroaralkyl;
R ⁵ , if present, is a hydrogen atom or alkyl;
E is ^{-Y bE -NR dE -CO-L E} - a, where in ^{Y bE} is _bound, be a C 1 -C ₆ alkylene or _C 2 -C ₆ alkenylene;
R ^dE represents a hydrogen _atom, a C 1 -C ₆ alkyl or _C 2 -C ₆ alkenyl; are substituted and ^{L E} is cycloalkyl, heterocycloalkyl, aryl which may be optionally substituted, optionally Which can be heteroaryl or heteroaralkyl.

  In the 25th aspect of the first aspect, which is also one aspect of the first, fourth to sixth, twelfth, thirteenth, seventeenth to twentieth, twenty-second, or twenty-third aspects of the first aspect, The compound is of formula (III) or (IV):

Indicated by
Where
Y ¹ is N, CH or CF;
K is

Chosen from;
R ³ is a hydrogen atom or methyl;
R ^y2 is a hydrogen atom, a halogen atom or C ₁ -C ₆ alkyl;
R ^y3 is a hydrogen atom, a halogen atom, C ₁ -C ₆ alkyl or C ₁ -C ₆ heteroalkyl;
R ^y4 is a hydrogen atom or a halogen atom, C ₁ -C ₆ alkyl or C ₁ -C ₆ heteroalkyl;
E is ^{-Y aE -NR cE -CO-R dE} -, - Y aE -NR cE -CO-L E -, - Y aE -NR cE -CO-NR dE -L E - or ^-Y aE ^{-NR cE -CS-NR} dE -L ^E
-Where Y ^aE is a bond or C ₁ -C ₆ alkylene;
R ^cE is a hydrogen atom, C ₁ -C ₆ alkyl or C ₂ -C ₆ alkenyl;
R ^dE is a hydrogen atom, an optionally substituted C ₁ -C ₈ alkyl, an optionally substituted C ₂ -C ₈ alkenyl or an optionally substituted C _2- A C ₈ alkynyl group;
L ^E is cycloalkyl, heterocycloalkyl, alkylcycloalkyl, heteroaryl alkyl cycloalkyl, optionally aryl which may be substituted, optionally heteroaryl may be substituted, aralkyl or heteroaralkyl;
G, if present, is cycloalkyl, —Y ^aG —CO—NR ^{aGR bG} , —Y ^aG —O—R ^eG , —Y ^aG —S—R ^eG , —Y ^aG —SO—R ^eG or — Y ^aG —SO ₂ —R ^eG where Y ^aG is a bond or C ₁ -C ₆ alkylene;
R ^aG is a hydrogen atom or C ₁ -C ₆ alkyl;
R ^bG is a hydrogen atom or C ₁ -C ₆ alkyl; and R ^eG is C ₁ -C ₆ alkyl.

  In a twenty-sixth embodiment of the first aspect, which is also an embodiment of the twenty-fifth embodiment of the first aspect, the compound has the formula (V):

Indicated by
^{Wherein, Y 1, K, R 3} , R y2, R y3, R y4 and ^{L E} are as defined in the twenty-fifth aspect of the first aspect.

  In a twenty-seventh embodiment of the first aspect, which is also an embodiment of the twenty-fifth embodiment of the first aspect, the compound has the formula (VI):

Indicated by
^{Wherein, Y 1, K, R 3} , R y2, R y3, R y4 and ^{L E} are as defined in the twenty-fifth aspect of the first aspect.

In the twenty-eighth aspect of the first aspect, which is also one aspect of the twenty-fourth to twenty-seventh aspects of the first aspect,
R ^y3 is a hydrogen atom.

In the twenty-ninth aspect of the first aspect, which is also one of the twenty-fourth to twenty-eighth aspects of the first aspect,
R ^y2 is a hydrogen atom.

  Of the first side surface which is also one aspect of the first, fourth to sixth, ninth, tenth, twelfth, thirteenth, seventeenth, twenty-first, twenty-second, twenty-third or twenty-third aspects of the first side surface. In a thirtieth embodiment, the compound has the formula (VII):

Indicated by
Where
Y ¹ is N, CH or CF;
K is

Chosen from;
R ^y4 is a hydrogen atom, a halogen atom, C ₁ -C ₆ alkyl or C ₁ -C ₆ heteroalkyl;
E is ^-Y aE ^{-CO-L E} - a, where in ^{Y aE} is a bond or _C 1 -C ₆ alkylene;
L ^E is heterocycloalkyl; and G is an alkyl cycloalkyl, heterocycloalkylalkyl, aryl or heteroaryl ares.

In the thirty-first aspect of the first aspect, which is also one aspect of the thirty-first aspect of the first aspect, R ^y4 is a hydrogen atom or a halogen atom.

In the thirty-second aspect of the first aspect, which is also one aspect of the first to thirty-first aspects of the first aspect,
The stereogenic center to which R ³ is attached is in the (R) configuration; and R ³ is methyl.

In a thirty third embodiment of the first aspect which is also an embodiment of the first to thirty second aspects of the first aspect, the compound is 500 nM or more in a standard in vitro BK B1 receptor-mediated assay. A low IC ₅₀ is indicated.

  In the first embodiment of the second aspect, the problem underlying the present application is solved by a compound which is preferably a compound according to any one of the first to thirty-third embodiments of the first aspect, and the compound Is selected from compounds 1 to 282 of Table 1.

  In the first embodiment of the third aspect, the problem underlying the present application is one or more compounds according to the embodiment of any of the first and second aspects, optionally at least one carrier material, Solved by a pharmaceutical composition comprising excipients and / or auxiliaries.

  In a second embodiment of the third aspect, which is also an embodiment of the first embodiment of the third aspect, the pharmaceutical composition is an aerosol, cream, gel, pill, capsule, syrup, solution, transdermal therapeutic system Prepared as a suppository or pharmaceutical device.

  In the first embodiment of the fourth aspect, the problem underlying the present application is the embodiment of any of the first to third aspects for the manufacture of a medicament for the treatment and / or prevention of a disease or condition Solved by the use of a compound or pharmaceutical composition according to.

  In a second embodiment of the fourth aspect, which is also an embodiment of the first embodiment of the fourth aspect, the condition or disease is responsive to BK B1R modulation.

  In the third embodiment of the fourth aspect, which is also an embodiment of the first or second embodiment of the fourth aspect, the condition or disease is selected from the group comprising inflammatory diseases, immune disorders and pain.

  In the fourth embodiment of the fourth aspect, which is also one embodiment of the third embodiment of the fourth aspect, the inflammatory disease or immune disorder is inflammatory bowel disease, rheumatoid arthritis, gouty arthritis, atherosclerotic artery Selected from the group comprising sclerosis and associated fibrotic conditions.

  In the fifth embodiment of the fourth aspect, which is also an embodiment of the third embodiment of the fourth aspect, the pain is visceral pain, neuropathic pain, complex regional pain syndrome CRPS and inflammation Selected from the group comprising sexual pain.

In the first embodiment of the fifth aspect, the problem underlying the present application is BK in a test tube.
Solved by a method of inhibiting the binding of DAKD, KD and DABK to the B1 receptor, which inhibits the binding of DAKD, KD and DABK to the BK B1 receptor to a detectable extent. Contacting with at least one compound according to any of the embodiments of the first to third aspects or a pharmaceutically acceptable salt, solvate or hydrate thereof under conditions and in an amount sufficient thereto. Comprising.

In the first embodiment of the sixth aspect, the problem underlying the present application is solved by a method for localizing or detecting a BK B1 receptor in a tissue, preferably a tissue section, in a test tube. ,that is:
(A) a sample of the tissue presumed to contain a BK B1 receptor, a compound according to any of the embodiments of the first to third aspects labeled to detectably, and a compound to the BK B1 receptor And b) detecting a compound bound to the BK B1 receptor, or detecting binding of the compound to the BK B1 receptor.

  In a second embodiment of the sixth aspect that is also an embodiment of the first embodiment of the sixth aspect, the compound is radiolabeled, fluorescent-labeled, or luminescent labeled, or Label with antibody.

  In a first embodiment of the seventh aspect, the problem underlying this application comprises the administration of a compound or pharmaceutical composition according to any of the embodiments of the first to third aspects, such as It is solved by a method for treating a patient in need of treatment.

  Those skilled in the art will recognize that any compound resulting from the combination of individual generic groups of formula (I) disclosed herein should be encompassed by the present invention. Will.

  The compounds shown in Table 1 below are embodiments of formula (I) according to the present invention.

The B1R antagonists shown herein have high activity on human B1R, ie, an inhibition constant (IC ₅₀ ) lower than 5 micromolar for competition with binding of labeled DAKD to human B1R, or very high activity on human B1R. I.e., an IC ₅₀ preferably lower than 50 nanomolar for competition with binding of labeled DAKD to human B1R. In certain embodiments, such antagonists are highly active on non-human species B1R, ie, rabbit B
An IC ₅₀ lower than 5 micromolar for competition with binding of labeled DAKD to 1R and cynomolgus monkey is shown.

The activity and more particularly the pharmacological activity of the B1R antagonist according to the invention can be assessed using a suitable in vitro assay. For example, via a radioligand binding assay, such as the assay shown in Example 24, the IC ₅₀ value of an antagonist according to the invention for B1R can be determined, thus the assay shown in Example 24 is a standard in vitro BK One embodiment of a B1R-mediated assay. The inhibitory effect of the B1R antagonists shown herein on B1R can be determined via a calcium mobilization assay, such as the assay shown in Example 25, for example.

Preferred compounds of the invention have an IC ₅₀ (half-maximal inhibitory concentration) of about 5 micromolar or lower in the above assay, more preferably about 500 nM or lower, or even 50 nM or even lower. IC _50, even with a certain _{IC 50} more more preferably from about 10nM to or lower than, or 1 nanomolar to or lower than that.

  The invention further provides, within another aspect, a pharmaceutical composition comprising at least one B1R modulator as described herein in combination with a physiologically acceptable carrier or excipient. To do. Also provided are methods of preparing such pharmaceutical compositions. Such compositions are particularly useful in the treatment of B1R-mediated diseases as described below.

  These and other aspects of the invention will become apparent upon reference to the following detailed description.

The compounds are generally described herein using standard nomenclature. With respect to compounds having asymmetric centers, it should be understood that all optical isomers and mixtures thereof are encompassed otherwise. Compounds having two or more asymmetric elements can also exist as a mixture of diastereomers. In addition, compounds having carbon-carbon double bonds may exist in Z- and E-forms, and all isomers of the compounds are included in the present invention unless otherwise stated. Where a compound exists in various tautomers, the listed compounds are not limited to any one tautomer, but are intended to include all tautomers. The listed compounds are further intended to include compounds in which one or more atoms are replaced by isotopes, ie atoms having the same atomic number but different mass numbers. By way of general example and without limitation, hydrogen isotopes include tritium and deuterium, and carbon isotopes include ¹¹ C, ¹³ C, and ¹⁴ C.

  A compound according to the formula set forth herein having one or more stereogenic centers has an enantiomeric excess of at least 50%. For example, such compounds can have an enantiomeric excess of at least 60%, 70%, 80%, 85%, 90%, 95% or 98%. Some embodiments of the compounds have an enantiomeric excess of at least 99%. It will be apparent that single enantiomers (optically active forms) can be obtained by asymmetric synthesis, synthesis from optically pure precursors or resolution of racemates. The racemate can be resolved by conventional methods such as, for example, crystallization in the presence of a resolving agent or chromatography using, for example, a chiral HPLC column.

Certain compounds are herein defined as variables such as A, R ¹ , R ^a , W, Y, Z (vari).
described using a general formula including Unless otherwise stated, each variable in such a formula is defined independently of any other variable, and any variable that occurs more than once in a formula is defined independently in each occurrence. The Thus, for example, where a group is shown to be substituted with 0-2 R ^* , the group can be unsubstituted or substituted with up to 2 R ^* groups, each occurrence R ^{* in} is independently selected from the definition of R ^* . Also, combinations of substituents and / or variables are permitted only if such combinations result in stable compounds, i.e., compounds that can be isolated, characterized, and examined for biological activity. obtain.

  A “pharmaceutically acceptable salt” of a compound disclosed herein is a human or animal tissue without excessive toxicity or carcinogenicity, and preferably without irritation, allergic reactions or other problems or complications. Acid or base salts generally considered in the art to be suitable for use in contact with. Such salts include inorganic and organic acid salts of basic residues such as amines and alkali or organic salts of acidic residues such as carboxylic acids.

Suitable pharmaceutical salts include hydrochloric acid, phosphoric acid, hydrobromic acid, malic acid, glycolic acid, fumaric acid, sulfuric acid, sulfamic acid, sulfanilic acid, formic acid, toluenesulfonic acid, methanesulfonic acid, benzenesulfonic acid, Ethanedisulfonic acid, 2-hydroxyethylsulfonic acid, nitric acid, benzoic acid, 2-acetoxybenzoic acid, citric acid, tartaric acid, lactic acid, stearic acid, salicylic acid, glutamic acid, ascorbic acid, pamoic acid, succinic acid, fumaric acid, maleic acid , Propionic acid, hydroxymaleic acid, hydroiodic acid, phenylacetic acid, alkanoic acid such as acetic acid, n is an integer from 0 to 4, i.e. HOOC- (CH ₂ ) Acid salts such as, but not limited to, _n- COOH. Similarly, pharmaceutically acceptable cations include, but are not limited to sodium, potassium, calcium, aluminum, lithium and ammonium. One of ordinary skill in the art will recognize additional pharmaceutically acceptable salts for the compounds presented herein. In general, a pharmaceutically acceptable acid or base salt can be synthesized from a parent compound that contains a basic or acidic moiety by conventional chemical methods. In essence, such salts are prepared by reacting the free acid or base form of these compounds with a stoichiometric amount of a suitable base or acid in water or an organic solvent, or in a mixture of the two. be able to. In general, the use of non-aqueous media such as ether, ethyl acetate, ethanol, isopropanol or acetonitrile is preferred.

  Each compound of formula (I) can exist as a hydrate, solvate or non-covalent complex, but this is not necessary. In addition, the various crystal forms and polymorphs are within the scope of the invention, as are the prodrugs of the compounds of formula (I) set forth herein.

  “Prodrugs” may not completely meet the structural requirements of the compounds presented herein, but may be modified in vivo following administration to a subject or patient, A compound which gives a compound of formula (I) as indicated in the specification. For example, a prodrug can be an acylated derivative of a compound shown herein. Prodrugs have a hydroxy, carboxy, amine or sulfhydryl group attached to any group that cleaves to form a free hydroxy, carboxy, amino or sulfhydryl group, respectively, when administered to a mammalian patient Is included. Examples of prodrugs include, but are not limited to, acetate, formate, phosphate and benzoate derivatives of alcohol and amine functional groups within the compounds shown herein. Prodrugs of the compounds shown herein can be made by modifying functional groups present in the compound in such a way that the modification is cleaved in vivo to yield the parent compound.

As used herein, “substituent” refers to a molecular moiety that is covalently bonded to an atom in a molecule of interest. For example, a “ring substituent” is a moiety such as a halogen, alkyl group, haloalkyl group, or other substituent described herein that is covalently bonded to an atom that is a ring member, preferably a carbon or nitrogen atom. Can be. The term “substituted” as used herein means that one or more hydrogens on a specified atom are replaced with those selected from the substituents indicated, provided that The normal valence of the atom being made cannot be exceeded, and the substitution results in a stable compound, i.e. a compound that can be isolated, characterized and examined for biological activity. When a substituent is oxo, ie = O, two hydrogens on the atom are replaced. An oxo group that is a substituent of an aromatic carbon atom results in the conversion of —CH— to —C (═O) — and loss of aromaticity. For example, a pyridyl group substituted by oxo is pyridone.

  The expression alkyl is preferably a saturated linear or branched hydrocarbon containing 1 to 20 carbon atoms, preferably 1 to 12 carbon atoms, more preferably 1 to 6 carbon atoms. A group such as methyl, ethyl, propyl, iso-propyl, n-butyl, iso-butyl, sec-butyl, tert-butyl, n-pentyl, n-hexyl, 2,2-dimethylbutyl or n-octyl. .

  The terms alkenyl and alkynyl refer to at least partially unsaturated linear or branched containing 2 to 20 carbon atoms, preferably 2 to 12 carbon atoms, more preferably 2 to 6 carbon atoms. A branched hydrocarbon group such as an ethenyl, allyl, acetylenyl, propargyl, isoprenyl or hex-2-enyl group. Preferably, the alkenyl group has 1 or 2, more preferably 1 double bond, and the alkynyl group has 1 or 2, more preferably 1 triple bond.

  Furthermore, the terms alkyl, alkenyl and alkynyl mean radicals in which one or more hydrogen atoms are each independently replaced by halogen atoms, preferably F or Cl, such as 2,2,2-trichloroethyl or Refers to a trifluoromethyl group.

  The expression heteroalkyl preferably represents one or more, preferably 1, 2 or 3 carbon atoms, independently of each other oxygen, nitrogen, phosphorus, boron, selenium, silicon or sulfur atoms, preferably Refers to an alkyl, alkenyl or alkynyl group substituted by oxygen, sulfur or nitrogen, eg heteroalkenyl, heteroalkynyl. The expression heteroalkyl preferably further denotes a carboxylic acid or a group derived from a carboxylic acid, such as acyl, acylalkyl, alkoxycarbonyl, acyloxy, acyloxyalkyl, carboxyalkylamide, alkylcarbamoylalkyl, alkylcarbamoyloxyalkyl, alkylureido Refers to alkyl or alkoxycarbonyloxy.

Examples of heteroalkyl groups are of the formula ^{-S-Y a -L, -S-} Y a -CO-NR a R b, -Y a -NR c -CO-NR a R b, -Y a -NR c - CO—O—R ^d , —Y ^a —NR ^c —CO—R ^d , —Y ^a —NR ^c —CO—NR ^d —L, —Y ^a —NR ^c —CS—NR ^d —L, —Y ^a —O—CO—NR ^a R ^b , —Y ^a —CO—NR ^a R ^b , —O—Y ^a —CO—NR ^a R ^b , —Y ^a —NR ^c —CO—L, —Y ^a —O ^{-CO-O-R c, -Y} a -O-CO-R c, -Y a -O-R c, -Y a -CO-L, -Y a -NR a R b, R c -S- Y ^a −, R ^a —N (R ^b ) —Y ^a —, R ^c —CO—Y ^a —, R ^c —O—CO—Y ^a —, R ^c —CO—O—Y ^a —, R ^{c -CO-N (R b) -Y} a ^{-, R a -N (R b} ) -CO-Y a -, R c -SO-Y a -, R c -SO 2 -Y a -, - Y a -NR c -SO 2 -NR a R b , —Y ^a —SO ₂ —NR ^a R ^b , —Y ^a —NR ^c —SO ₂ —R ^d , R ^a —O—CO—N (R ^b ) —Y ^a —, R ^a —N (R ^{b ) -C (= NR d) -N} (R c) -Y a -, R c -S-CO-Y a -, R c -CO-S-Y a -, R c -S-CO-N ( R ^b ) —Y ^a —, R ^a —N (R ^b ) —
CO—S—Y ^a —, R ^c —S—CO—O—Y ^a —, R ^c —O—CO—S—Y ^a —, R ^c —S—CO—S—Y ^a —. Where R ^a is a hydrogen atom, C ₁ -C ₆ alkyl, C ₂ -C ₆ alkenyl, C ₂ -C ₆ alkynyl, or is bonded to R ^b to form a 4- to 10-membered cycloalkyl or hetero; Cycloalkyl is formed; R ^b is a hydrogen atom, C ₁ -C ₆ alkyl, C ₂ -C ₆ alkenyl or C ₂ -C ₆ alkynyl, or together with R ^a 4-10-membered Forming a cycloalkyl or heterocycloalkyl; R ^c is a hydrogen atom, an optionally substituted C ₁ -C ₈ alkyl, an optionally substituted C ₂ -C ₈ alkenyl or optionally C that can be substituted _2- C ₈ alkynyl; R ^d is a hydrogen atom, optionally substituted C ₁ -C ₈ alkyl, optionally substituted C ₂ -C ₈ alkenyl or optionally substituted C ₂ -C ₈ alkynyl which can be; L is cycloalkyl, heterocycloalkyl, alkylcycloalkyl, heteroalkylcycloalkyl, optionally substituted aryl, optionally substituted And Y ^a is a bond, a C ₁ -C ₆ alkylene, a C ₂ -C ₆ alkenylene or a C ₂ -C ₆ alkynylene group; and each heteroalkyl group has at least one heteroaryl, aralkyl or heteroaralkyl Of carbon atoms and one or more hydrogen atoms are fluorine It is possible to have been replaced by chlorine atoms. Certain compounds are described herein using formulas that include variables having superscripts B, E, or G as extensions, eg, L ^B , Y ^aE, or R ^cG . Unless otherwise indicated, the definition of these variables correspond to the above definition of the respective variables, i.e., ^{L B} and ^{L E} is defined as ^{L; Y aB,} Y ^aE and ^{Y aG} is defined as ^{Y a} R ^cB , R ^cE and R ^cG are defined as R ^c and so on; Specific examples of heteroalkyl groups are methoxy, trifluoromethoxy, ethoxy, n-propyloxy, isopropyloxy, tert-butyloxy, methoxymethyl, ethoxymethyl, methoxyethyl, methylamino, ethylamino, dimethylamino, diethylamino, isopropyl Ethylamino, methylaminomethyl, ethylaminomethyl, diisopropylaminoethyl, enol ether, dimethylaminomethyl, dimethylaminoethyl, acetyl, propionyl, butyryloxy, acetyloxy, methoxycarbonyl, ethoxycarbonyl, isobutyrylamino-methyl, N- Ethyl-N-methylcarbamoyl and N-methylcarbamoyl. Further examples of heteroalkyl groups are nitrile, isonitrile, cyanate, thiocyanate, isocyanate, isothiocyanate and alkyl nitrile groups. Examples of heteroalkylene groups are groups of the formula —CH ₂ CH (OH) — or —CONH—.

The expression cycloalkyl is preferably 1 or more containing 3 to 14 ring carbon atoms, preferably 3 to 10, more preferably 3, 4, 5, 6 or 7 ring carbon atoms. Refers to a saturated or partially unsaturated cyclic group containing many rings, preferably one or two rings. In one embodiment, the partially unsaturated cyclic group has one, two or more double bonds, such as a cycloalkenyl group. The expression cycloalkyl preferably further means that one or more hydrogen atoms are each independently of the other by fluorine, chlorine, bromine or iodine atoms, or OH, ═O, SH, ═S, NH ₂ , = A group which is replaced by an NH, CN, CF ₃ or NO ₂ group, thus for example a cyclic ketone, such as cyclohexanone, 2-cyclohexenone or cyclopentanone. Yet another specific example of a cycloalkyl group is cyclopropyl, cyclobutyl, cyclopentyl, spiro [4,5] decanyl, norbornyl, cyclohexyl, cyclopentenyl, cyclohexadienyl, decalinyl, bicyclo [4.3.0] nonyl. , Tetralin, cyclopentylcyclohexyl, fluorocyclohexyl or cyclohex-2-enyl group.

The expression heterocycloalkyl is preferably one or more, preferably 1, 2 or 3 ring carbon atoms independently of each other oxygen, nitrogen, silicon, selenium, phosphorus or sulfur atoms, preferably Refers to a cycloalkyl group as defined above which is replaced by oxygen, sulfur or nitrogen. Heterocycloalkyl groups preferably have 1 or 2 rings containing 3 to 10, more preferably 3, 4, 5, 6 or 7 ring atoms. The expression heterocycloalkyl preferably further means that one or more hydrogen atoms are each independently of the other by fluorine, chlorine, bromine or iodine atoms, or OH, ═O, SH, ═S, NH ₂ , = Refers to a group that is replaced by a NH, CN, CF ₃ or NO ₂ group. Examples are piperidyl, piperazinyl, morpholinyl, urotropinyl, pyrrolidinyl, tetrahydrothiophenyl, tetrahydropyranyl, tetrahydrofuryl or 2-pyrazolinyl groups and also lactams, lactones, cyclic imides and cyclic anhydrides.

  The expression alkylcycloalkyl is preferably a group containing both cycloalkyl and also alkyl, alkenyl or alkynyl groups according to the above definition, for example alkylcycloalkyl, cycloalkylalkyl, alkylcycloalkenyl, alkenylcycloalkyl and alkynylcycloalkyl. Refers to an alkyl group. Alkylcycloalkyl groups are preferably cycloalkyl groups containing 1 or 2 ring systems having 3 to 10, preferably 3, 4, 5, 6 or 7 carbon atoms and 1 or 2 to 6 Containing 1 or 2 alkyl, alkenyl or alkynyl groups having 1 carbon atom, the cyclic group can be optionally substituted.

  The expression heteroalkylcycloalkyl is preferably one or more, preferably 1, 2 or 3 carbon atoms independently of each other oxygen, nitrogen, silicon, selenium, phosphorus or sulfur atoms, preferably Refers to an alkylcycloalkyl group as defined above which is replaced by oxygen, sulfur or nitrogen. A heteroalkylcycloalkyl group preferably contains 1 to 2 ring systems having 3 to 10, preferably 3, 4, 5, 6 or 7 ring atoms and 1 or 2 to 6 carbon atoms. Contains 1 or 2 alkyl, alkenyl, alkynyl or heteroalkyl groups having. Examples of such groups are alkylheterocycloalkyl, heterocycloalkylalkyl, alkylheterocycloalkenyl, alkenylheterocycloalkyl, alkynylheterocycloalkyl, heteroalkylcycloalkyl, heteroalkylheterocycloalkyl and heteroalkylheterocycloalkenyl. Yes, the cyclic group can be optionally substituted and can be saturated or mono-, di- or tri-unsaturated.

The expression aryl or Ar preferably represents an aromatic containing one or more rings containing 6 to 14 ring carbon atoms, preferably 6 to 10 and more preferably 6 ring carbon atoms. Refers to the group. The expression aryl (or Ar) preferably further means that one or more hydrogen atoms are each independently of the other by fluorine, chlorine, bromine or iodine atoms, or OH, SH, NH ₂ , CN, CF ₃ Or refers to a group replaced by a NO ₂ group. Examples are the phenyl, naphthyl, biphenyl, 2-fluorophenyl, anilinyl, 3-nitrophenyl or 4-hydroxyphenyl groups.

The expression heteroaryl preferably contains 5 to 14 ring atoms, preferably 5 to 10, more preferably 5 or 6 ring atoms, and one or more, preferably 1, Refers to an aromatic group containing one or more rings containing 2, 3 or 4 oxygen, nitrogen, phosphorus or sulfur ring atoms, preferably O, S or N. The expression heteroaryl preferably further means that one or more hydrogen atoms are each independently of the other by fluorine, chlorine, bromine or iodine atoms, or OH, ═O, SH, NH ₂ , ═NH, CN , Refers to a group that is replaced by a CF ₃ or NO ₂ group. Examples are 4-pyridyl, 2-imidazolyl, 3-phenylpyrrolyl, thiazolyl, oxazolyl, triazolyl, tetrazolyl, isoxazolyl, indazolyl, indolyl, benzimidazolyl, pyridazinyl, quinolinyl, purinyl, carbazolyl, acridinyl, pyrimidyl, 2,3'- Bifuryl, 3-pyrazolyl and isoquinolinyl.

  The expression aralkyl is preferably a group containing both aryl and also alkyl, alkenyl, alkynyl and / or cycloalkyl groups as defined above, eg arylalkyl, arylalkenyl, arylalkynyl, arylcycloalkyl, arylcycloalkenyl. , Alkylarylcycloalkyl and alkylarylcycloalkenyl groups. Specific examples of aralkyl are toluene, xylene, mesitylene, styrene, benzyl chloride, o-fluorotoluene, 1H-indene, tetralin, dihydronaphthalene, indanone, phenylcyclopentyl, cumene, cyclohexylphenyl, fluorene and indane. Aralkyl groups are preferably 1 or 2 aromatic ring systems, 1 or 2 rings containing 6 to 10 carbon atoms and 1 or 2 containing 1 or 2 to 6 carbon atoms. Containing alkyl, alkenyl and / or alkynyl groups and / or cycloalkyl groups containing 5 or 6 ring carbon atoms.

  The expression heteroaralkyl preferably represents one or more, preferably 1, 2, 3 or 4 carbon atoms, independently of each other, oxygen, nitrogen, silicon, selenium, phosphorus, boron or sulfur atoms, Aralkyl groups as defined above which are preferably replaced by oxygen, sulfur or nitrogen, ie aryl or heteroaryl as defined above and also alkyl, alkenyl, alkynyl and / or heteroalkyl and / or cycloalkyl and / or hetero Refers to a group containing both cycloalkyl groups. Heteroaralkyl groups preferably contain 1 or 2 aromatic ring systems, 1 or 2 rings containing 5 or 6 to 10 carbon atoms and 1 or 2 to 6 carbon atoms Containing 1 or 2 alkyl, alkenyl and / or alkynyl groups and / or cycloalkyl groups containing 5 or 6 ring carbon atoms, wherein 1, 2, 3 or 4 of these carbon atoms are each Independently replaced by oxygen, sulfur or nitrogen atoms.

  Examples of heteroaralkyl groups are arylheteroalkyl, arylheterocycloalkyl, arylheterocycloalkenyl, arylalkylheterocycloalkyl, arylalkenylheterocycloalkyl, arylalkynylheterocycloalkyl, arylalkylheterocycloalkenyl, heteroarylalkyl, hetero Arylalkenyl, heteroarylalkynyl, heteroarylheteroalkyl, heteroarylcycloalkyl, heteroarylcycloalkenyl, heteroarylheterocycloalkyl, heteroarylheterocycloalkenyl, heteroarylalkylcycloalkyl, heteroarylalkylheterocycloalkenyl, heteroarylhetero Alkylcycloalkyl, heteroarylheteroalkyl Cycloalkenyl, heteroaryl alkyl heteroarylalkyl and heteroaryl heteroalkyl heterocycloalkyl group, cyclic group or mono- saturated -, di - unsaturated - or birds. Specific examples are tetrahydroisoquinolinyl, benzoyl, 2- or 3-ethylindolyl, 4-methylpyridino, 2-, 3- or 4-methoxyphenyl, 4-ethoxyphenyl, 2-, 3- or 4 -A carboxyphenylalkyl group.

The expressions cycloalkyl, heterocycloalkyl, alkylcycloalkyl, heteroalkylcycloalkyl, aryl, heteroaryl, aralkyl and heteroaralkyl are such that one or more hydrogen atoms of such groups are each independently fluorine. , A chlorine, bromine or iodine atom or a group which is replaced by an OH, ═O, SH, ═S, NH ₂ , ═NH, CN, CF ₃ or NO ₂ group.

The expression “optionally substituted” used in connection with any group is one or more hydrogen atoms independently of each other by fluorine, chlorine, bromine or iodine atoms, or OH, ═O, SH, ═S, NH ₂ , ═NH, CN, CF ₃ or refers to a group that is replaced by a NO ₂ group. The expression further includes one or more hydrogen atoms, each independently of the other, unsubstituted C ₁ -C ₆ alkyl, unsubstituted C ₂ -C ₆ alkenyl, unsubstituted C ₂ -C ₆ alkynyl, unsubstituted C ₁ -C ₆ heteroalkyl, unsubstituted _C 3 _{-C 10} cycloalkyl, unsubstituted _C 2 -C ₉ heterocycloalkyl, unsubstituted _C 6 _{-C 10} aryl, unsubstituted _C 1 -C ₉ heteroaryl, unsubstituted C It refers to ₇ -C ₁₂ aralkyl or unsubstituted _C 2 _{-C 11} group substituted by heteroaralkyl group.

  The expression “halogen” preferably used herein means fluorine, chlorine, bromine, iodine.

  As used herein, terms that define the limits of length ranges, such as “1-5”, mean integers from 1 to 5, ie 1, 2, 3, 4, and 5. In other words, a range defined by two explicitly stated integers shall comprise and disclose an integer that defines the limit and an integer contained within the range.

  Therapeutic uses of the compounds of formula (I), their pharmacologically acceptable salts, solvates and hydrates and also the formulations and pharmaceutical compositions containing them are within the scope of the present invention. is there. The invention also relates to the use of those compounds of formula (I) as active ingredients in the preparation or manufacture of a medicament.

The pharmaceutical composition according to the invention comprises at least one compound of formula (I) and optionally one or more carrier substances, excipients and / or auxiliaries. The pharmaceutical composition further comprises, for example, water, buffering agents such as neutral buffered saline or phosphate buffered saline, ethanol, mineral oil, vegetable oil, dimethyl sulfoxide, carbohydrates such as glucose, mannose, sucrose or dextran, mannitol, It may comprise one or more of proteins, auxiliaries, polypeptides or amino acids such as glycine, antioxidants, chelating agents such as EDTA or glutathione and / or preservatives. In addition, one or more other active ingredients can be included in the pharmaceutical compositions provided herein, but are not required. For example, the compounds of the present invention may be combined with antibiotics, anti-fungal agents, anti-viral agents, anti-histamines, non-steroidal anti-inflammatory agents, disease modifying anti-rheumatic agents.
drug), a cytostatic drug, a drug having smooth muscle activity-regulating activity, or a mixture thereof can be advantageously used.

  Any suitable including, for example, topical, eg transdermal or ocular, oral, buccal, nasal, vaginal, rectal or parenteral administration Pharmaceutical compositions can be prepared for the route of administration. The term parenteral as used herein refers to subcutaneous, intradermal, intravascular, eg, intravenous, intramuscular, spinal, intracranial, intradural, intraocular, periocular, orbital Internal, intrasynovial and intraperitoneal injections and any similar injection or infusion method. In certain embodiments, compositions in a form suitable for oral use are preferred. Such forms include, for example, tablets, troches, lozenges, aqueous or oily suspensions, dispersible powders or granules, emulsions, hard or soft capsules or syrups or elixirs. Within yet another aspect, the compositions provided herein can be prepared as a lyophilizate. For certain conditions, such as in the treatment of skin conditions such as burns or itching, topical formulations may be preferred.

Compositions intended for oral use further comprise one or more ingredients such as sweeteners, flavorings, colorants and / or preservatives to provide attractive and palatable preparations. Can be Tablets contain the active ingredient in admixture with physiologically acceptable excipients that are suitable for the manufacture of tablets. Such excipients include, for example, inert diluents such as calcium carbonate, sodium carbonate, lactose, calcium phosphate or sodium phosphate, granulating and disintegrating agents such as corn starch or alginic acid, binders such as starch, gelatin or Gum arabic and lubricants such as magnesium stearate, stearic acid or talc are included. The tablets can be uncoated or they can be coated in a known manner to delay disintegration and absorption in the gastrointestinal tract and thereby provide a sustained action over a longer period. For example, a time delay material such as glyceryl monostearate or glyceryl distearate can be used.

  Formulations for oral use include hard gelatin capsules in which the active ingredient is mixed with an inert solid diluent such as calcium carbonate, calcium phosphate or kaolin or the active ingredient is water or an oil medium such as peanut oil, liquid paraffin or olive oil It can also be provided as a soft gelatin capsule to be mixed.

  Aqueous suspensions contain the active ingredients in admixture with excipients suitable for the manufacture of aqueous suspensions. Such excipients include suspending agents such as sodium carboxymethylcellulose, methylcellulose, hydropropylmethylcellulose, sodium alginate, polyvinylpyrrolidone, gum tragacanth and gum arabic; and dispersing or wetting agents such as naturally occurring phosphatides such as Condensation products of lecithin, alkylene oxide and fatty acids, such as polyoxyethylene stearate, condensation products of ethylene oxide and long chain aliphatic alcohols, such as heptadecaethyleneoxycetanol, condensation of partial esters derived from ethylene oxide and fatty acids and hexitol Products such as polyoxyethylene sorbitol monooleate or condensation formation of partial esters derived from ethylene oxide and fatty acids and anhydrous hexitol , It includes, for example, polyethylene sorbitan monooleate. Aqueous suspensions are one or more preservatives such as ethyl or n-propyl p-hydroxybenzoate, one or more colorants, one or more flavors and one or more Sweeteners such as sucrose or saccharin can also be included.

  Oily suspensions may be prepared by suspending the active ingredient in a vegetable oil, for example arachis oil, olive oil, sesame oil or coconut oil, or in a mineral oil such as liquid paraffin. Oily suspensions may contain a thickening agent such as beeswax, hard paraffin or cetyl alcohol. Sweetening agents such as those set forth above, and / or flavoring agents can be added to provide an oral suspension that is palatable. Such suspending agents can be preserved by the addition of an antioxidant such as ascorbic acid.

  Dispersible powders and granules suitable for preparation of an aqueous suspension by the addition of water provide the active ingredient in admixture with a dispersing or wetting agent, suspending agent and one or more preservatives. Suitable dispersing or wetting agents and suspending agents are represented by those already mentioned above. Additional excipients such as sweetening, flavoring and coloring agents can also be present.

  The pharmaceutical composition can also be in the form of an oil-in-water emulsion. The oily phase can be a vegetable oil, for example olive oil or arachis oil, a mineral oil, for example liquid paraffin or mixtures of these. Suitable emulsifiers include naturally occurring gums such as gum arabic or tragacanth, naturally occurring phosphatides such as soy lecithin and fatty acids and hexitol, esters or partial esters derived from anhydrides such as sorbitan monooleate and fatty acids. Condensation products of partial esters derived from hexitol and ethylene oxide, such as polyoxyethylene sorbitan monooleate, are included. Emulsions can also comprise one or more sweeteners and / or flavoring agents.

  Syrups and elixirs can be prepared with sweetening agents, for example glycerol, propylene glycol, sorbitol or sucrose. Such formulations can also comprise one or more demulcents, preservatives, flavoring agents and / or coloring agents.

The compounds can be prepared for local or topical administration, for example for topical application to the mucosa such as in the skin or eye. Formulations for topical administration typically comprise a topical vehicle combined with an active agent with or without additional optional ingredients. Suitable topical vehicles and additional components are well known in the art, and it will be apparent that the choice of vehicle will depend on the particular physical form and mode of delivery. Topical vehicles include water; organic solvents such as alcohols such as ethanol or isopropyl alcohol or glycerine; glycols such as butylene, isoprene or propylene glycol; aliphatic alcohols such as lanolin; mixtures of water and organic solvents and alcohols and glycerin Mixtures of organic solvents; lipid-based materials such as fatty acids, acylglycerols including oils, such as mineral oils and fats, phosphoglycerides, sphingolipids and waxes of natural or synthetic origin; protein-based materials such as collagen and gelatin; Included are both volatile and volatile silicone based materials; and hydrocarbon based materials such as microsponges and polymer matrices. The composition further comprises one or more ingredients adapted to improve the stability or effectiveness of the applied formulation, such as stabilizers, suspending agents, emulsifiers, viscosity modifiers, gelling agents, Preservatives, antioxidants, skin penetration enhancers, humectants and sustained release materials can be included. Examples of such ingredients include Martindale The Extra Pharmacopoeia (Pharmaceutical
Press, London 1993) and Martin (ed.), Remington's Pharmaceutical Sciences. The formulation can comprise microcapsules such as hydroxymethylcellulose or gelatin-microcapsules, liposomes, albumin microspheres, microemulsions, nanoparticles or nanocapsules.

  Topical preparations can be prepared in a variety of physical forms including, for example, solids, pastes, creams, foams, lotions, gels, powders, aqueous liquids, emulsions, sprays and skin patches. The presence of emulsifiers and viscosity modifiers and their amounts present in the formulation can dominate the physical appearance and viscosity of such forms. Solids are generally hard and cannot be poured, usually prepared as sticks or sticks, or in particulate form; solids can be opaque or transparent, optionally with solvents, emulsifiers, humectants, softeners, Fragrances, dyes / colorants, preservatives and other active ingredients that increase or enhance the effectiveness of the final product can be included. Creams and lotions are often similar to each other and differ primarily in their viscosity; both lotions and creams can be opaque, translucent or transparent, often emulsifiers, solvents and viscosity modifiers Moisturizers, softeners, fragrances, dyes / colorants, preservatives and other active ingredients that increase or enhance the effectiveness of the final product may be included. Gels with a range of viscosities from rich or high viscosity to dilute or low viscosity can be prepared. These formulations, as well as those in lotions and creams, increase or enhance the effectiveness of solvents, emulsifiers, humectants, softeners, fragrances, dyes / colorants, preservatives and the final product. Active ingredients. Liquids are thinner than creams, lotions or gels and often do not contain emulsifiers. Liquid topical products are often solvents, emulsifiers, humectants, softeners, fragrances, dyes / colorants, preservatives and other activities that increase or enhance the effectiveness of the final product. Contains ingredients.

Suitable emulsifiers for use in topical preparations include ionic emulsifiers, cetearyl alcohols, non-ionic emulsifiers such as polyoxyethylene oleyl ether, PEG-40 stearate, cetealess-12, cetealess-20, Includes, but is not limited to, cetealess-30, cetealess alcohol, PEG-100 stearate and glyceryl stearate. Suitable viscosity modifiers include, but are not limited to, protective colloids or non-ionic rubbers such as hydroxyethylcellulose, xanthan gum, magnesium aluminum silicate, silica, microwax, beeswax, paraffin and cetyl palmitate. . By adding a gelling agent such as chitosan, methylcellulose, ethylcellulose, polyvinyl alcohol, polyquaterniums, hydroxyethylcellulose, hydroxypropylcellulose, hydroxypropylmethylcellulose, carbomer or ammoniated glycyrrhizinate Can be formed. Suitable surfactants include, but are not limited to, nonionic, amphoteric, ionic and anionic surfactants. For example, one or more of dimethicone copolyol, polysorbate 20, polysorbate 40, polysorbate 60, polysorbate 80, lauramide DEA, cocamide DEA and cocamide MEA, oleyl betaine, cocamidopropylphosphatidyl PG-dimonium chloride and ammonium laureth sulfate It can be used in topical formulations.

  Suitable preservatives include antimicrobials such as methylparaben, propylparaben, sorbic acid, benzoic acid and formaldehyde and physical stabilizers and antioxidants such as vitamin E, sodium ascorbate / ascorbic acid and gallic acid Including but not limited to propyl. Suitable humectants include, but are not limited to, lactic acid and other hydroxy acids and their salts, glycerin, propylene glycol and butylene glycol. Suitable softeners include lanolin alcohol, lanolin, lanolin derivatives, cholesterol, petrolatum, isostearyl neopentanoate and mineral oil. Suitable fragrances and colorants include FD & C Red No. 40 and FD & C Yellow No. 5 is included, but is not limited to these. Other suitable additional ingredients that can be included in the topical formulations include abrasives, absorbents, anti-caking agents, antifoaming agents, antistatic agents, astringents such as, for example, hazel hazel, alcohol And herbal extracts such as chamomile extracts, binders / excipients, buffers, chelators, film formers, conditioning agents, propellants, opacifiers, pH adjusters and protectants Including, but not limited to.

  Examples of suitable topical vehicles for the preparation of gels are: hydroxypropylcellulose (2.1%); 70/30 isopropyl alcohol / water (90.9%); propylene glycol (5.1%) and polysorbate 80 (1.9%). Examples of suitable topical vehicles for foam preparation are: cetyl alcohol (1.1%); stearyl alcohol (0.5%); quaternium 52 (1.0%); propylene glycol (2.0% ); Ethanol 95 PGF3 (61.05%); deionized water (30.05%); P75 hydrocarbon propellant (4.30%). All percentages are by weight.

  Typical modes of delivery for topical compositions include finger application; application using a physical applicator such as cloth, tissue, cotton wool, stick or brush; mist, aerosol or foam spray spray Spray spraying including: dropper application; spraying; dipping; and rinsing. Controlled release vehicles can also be used and compositions can be prepared for transdermal administration as transdermal patches.

The pharmaceutical composition can be prepared as an inhalation preparation containing a spray, mist or aerosol. Such formulations are particularly useful for the treatment of asthma and other respiratory conditions. In the case of inhalation formulations, the compounds shown herein can be delivered via any inhalation method known to those skilled in the art. Such inhalation methods and devices include, but are not limited to, metered dose inhalers that use propellants such as CFC or HFA or physiologically and environmentally acceptable propellants. Absent. Other suitable devices are breath manipulated inhalers, multidose dry powder inhalers and aerosol nebulizers. Aerosol formulations for use in the present method typically include propellants, surfactants and cosolvents and can be filled into a conventional aerosol container that is closed by a suitable metering valve.

  Inhalation compositions can comprise liquid or powder compositions containing active ingredients suitable for nebulization and intrabronchial use or aerosol compositions administered via aerosol devices that dispense metered doses. . Suitable liquid compositions comprise the active ingredient in an aqueous pharmaceutically acceptable inhalant solvent such as isotonic saline or bacteriostatic water. The solution may be pumped or squeezed-actuated nebulized spray dispenser, or to allow or enable the required dosage of the liquid composition into the patient's lungs. It is administered by other conventional means. When the carrier is a liquid, preparations suitable for administration, for example as a nasal spray or as a nasal spray, include aqueous or oily solutions of the active ingredients.

  When the carrier is a solid, formulations or compositions suitable for nasal administration include, for example, a coarse powder having a particle size in the range of 20 to 500 microns, that is, in the manner in which the insufflation agent is administered, ie Administered by rapid inhalation through a nasal passage from a container of powder held in close contact with the nose. Suitable powder compositions include, by way of example, powder preparations of the active ingredient intimately mixed with lactose or other inert powders acceptable for intrabronchial administration. In a breakable capsule that can be inserted by a patient into a device in which the powder composition is administered via an aerosol dispenser, or the capsule is pierced and the powder is blown in a flow suitable for inhalation Can be put in.

  The pharmaceutical composition can also be prepared in the form of suppositories, eg for rectal administration. By mixing the drug with a suitable non-irritating excipient that is solid at ambient temperature but liquid at rectal temperature and will therefore melt in the rectum to release the drug, such as A composition can be prepared. Suitable excipients include, for example, cocoa butter and polyethylene glycols.

  The pharmaceutical composition can be prepared as a sustained release formulation, eg, a capsule-like formulation that results in a slow release of the modulator following administration. Such formulations are generally prepared using well-known methods and can be administered, for example, by oral, rectal or subcutaneous implantation, or by implantation at the desired target site. Carriers for use in such formulations are biocompatible and can also be biodegradable; preferably the formulations provide a relatively constant level of modulator release. The amount of modulator contained in the sustained release formulation depends, for example, on the site of implantation, the rate of release and the expected duration and the nature of the condition to be treated or prevented.

For the prevention and / or treatment of diseases mediated by BK or the like, the dosage of the biologically active compound according to the invention can vary within wide limits and can be adapted to individual requirements. . The active compounds according to the invention are generally administered in a therapeutically effective amount. Preferred dosages range from about 0.1 mg to about 140 mg per kilogram of body weight per day and from about 0.5 mg to about 7 g per patient per day. The daily dosage can be administered as a single dosage or in multiple dosages. The amount of active ingredient that can be combined with the carrier materials to produce a single dosage form will vary depending upon the host treated and the particular mode of administration. Dosage unit forms will generally contain about 1 mg to about 500 mg of an active ingredient.

  However, the specific dosage level for any particular patient will depend on the activity, age, weight, general health, sex, diet, time of administration, route of administration and rate of excretion of the particular compound used, It will be appreciated that the combination will depend on a variety of factors, including the other drugs being used for the treatment of the patient and the severity of the particular disease being treated.

  Preferred compounds of the present invention will have certain pharmacological properties. Such properties include, but are not limited to, oral bioavailability so that the preferred oral dosage forms discussed above can provide therapeutically effective levels of compounds in vivo.

  The B1R antagonist of formula (I) according to the present invention can be suitably used as an antagonist of B1R in a variety of applications both in vitro and in vivo. A B1R antagonist according to the present invention can be used to inhibit the binding of a BK B1R ligand, such as DAKD, to B1R in vitro or in vivo. The BK B1R antagonists shown herein are preferably administered orally or topically to a patient, such as a human, and are present in at least one body fluid or tissue of the patient while modulating BK B1R activity. . Accordingly, the present invention further provides a method for treating a patient suffering from a condition responsive to the B1R modulation referred to above. As used herein, the term “treatment” includes both disease-modifying treatment and treatment of symptoms, both of which prevent, delay or reduce the severity of symptoms. That is, before the onset of symptoms, or therapeutic to reduce the severity and / or duration of the symptoms, ie after the onset of symptoms. A condition is “responsive to B1R modulation” if modulation of B1R activity results in alleviation or healing of the condition or its symptoms. Patients may be administered as described herein with primates, particularly humans, domesticated companion animals such as dogs, cats, horses and livestock such as cows, pigs, Sheep can be included, but are not limited to these.

  Since various diseases involve the bradykinin B1 receptor, the B1 antagonist according to the present invention is a condition or disease responsive to BK B1R modulation, as is apparent from the prior art and in particular from the references cited herein. It is effective in the treatment and / or prevention. Animal models of these diseases are generally well known in the art and the compounds of the present invention can be used to evaluate their potential utility. Alternatively, the compounds according to the present invention are useful in other applications including, but not limited to, the diagnosis of such diseases and their use as in vivo and in vitro research tools.

  The compounds of the invention are antagonists of BK B1R and are therefore useful in the treatment and prevention of diseases and conditions mediated through the BK receptor pathway, such as inflammation, immune disorders and pain. The compounds are preferably effective in the treatment or prevention of inflammation such as but not limited to persistent or chronic inflammatory diseases, immune disorders, autoimmune diseases, neurogenic inflammation, inflammation-related edema and fibrosis. This includes, but is not limited to, gastrointestinal inflammation, septic shock, skin disease, respiratory tract disease and angiopathy. The compounds are preferably also effective in the treatment or prevention of pain such as chronic pain, inflammatory pain, visceral pain and neuropathic pain. This includes, but is not limited to, complex regional pain syndrome (CRPS).

It is within the scope of the present invention to use a compound according to the present invention as a diagnostic or for the manufacture thereof, wherein such a diagnostic is used for therapeutic purposes as disclosed herein. For the diagnosis of diseases and conditions to which the compounds of the invention can face.

  The following further defines various diseases and conditions that are responsive to B1R modulation and the use of the compounds according to the invention in certain methods and diagnostics.

Inflammatory Diseases and Immune Disorders Within this application, the term “inflammatory disease” preferably refers to acute-phase reaction, local and systemic inflammation and other forms of pathogenesis or pathogenesis. Including, but not limited to, disorders such as inflammation caused by disease and caused by inflammatory diseases as defined within this application.

  Within this application, the term “immune disorder” preferably refers to hypersensitivity, autoimmune disorders, transplant rejection in transplants, transplant toxicity, granulitis / tissue remodeling, myasthenia gravis, immunosuppression , Immune complex diseases, antibody over- and under-production and disorders such as vasculitis, but are not limited to these.

Since various diseases involve the bradykinin B1 receptor, the B1 antagonists according to the present invention are preferably treated / treated for conditions or diseases responsive to BK B1R modulation, as will be apparent from among the references cited herein, among others. Effective in prevention. Therefore, the compounds according to the present invention are suitable for inflammatory bowel diseases including Crohn's disease and ulcerative colitis (Stadnicki et al., Am. J. Physiol. Gastrointest River Physiol. 289 (2), 2005, G361-6; et al., Am. J. Gastroeneol, 97 (8), 2002, 2026-32; Devani et al., Dig. Liv.Disease, 37 (9), 2005, 665-73), hypersensitivity. It is effective in the treatment / prevention of inflammatory diseases such as but not limited to bowel syndrome, enteritis, liver disease, pancreatitis, gallbladder disease, smooth muscle relaxation for the treatment of gastrointestinal or uterine spasms. They can also be used in kidney diseases such as nephritis, bladder diseases such as cystitis, interstitial cystitis, ocular diseases such as uveitis, retinitis, glaucoma and otic diseases such as otitis media . They can also be used in inflammatory skin diseases such as psoriasis, eczema, atopic diseases, dermatitis and itching. The compounds may be used in joint and bone diseases such as juvenile or adult onset rheumatoid arthritis and gouty arthritis (Cassim).
et al. Author, Pharmacol. Ther. 94, 2002, 1-34; Sharma et al. Author, Exp. Toxic Pathol. 46, 1994, 421-433; Brechter et al. Author, Arthr. Rheum. 56 (3), 2007, 910-923), ankylosing spondylitis, adult-onset or systemic-onset childhood-onset juvenile idiopathic arthritis, psoriasis, psoriasis, psoriasis Effective in arthritis and osteoarthritis. The compounds are also effective in edema associated with burns, sprains or fractures, brain edema, closed head trauma and angioedema. They are immune disorders such as diabetic angiopathy, type I diabetes, diabetic nephropathy, diabetic neuropathy, diabetic retinopathy, diabetic angiopathy, post-capillary resistance or insulits. ) Associated with diabetes syndromes such as hyperglycemia, diuresis, proteinuria and increased urinary excretion of nitrite and kallikrein. Furthermore, they can be used in CNS disorders such as multiple sclerosis, epilepsy, amyotrophic lateral sclerosis, Alzheimer's disease, stroke, Parkinson's disease. They can also be used in cardiovascular diseases such as congestive heart failure, myocardial infarction, systemic inflammatory response syndrome (SIRS), ischemia-reperfusion injury and atherosclerosis (Raidoo et al , Immunopharmacol, 36 (2-3), 1997, 153-60; McLean et al., Cardiovasc. Res. 48, 2000, 194-210). Vascular disorders such as vasculitis, septic shock, anticirculating plasma volume reduction and / or antihypertensive drugs, closed head trauma, cancer, sepsis, gingivitis, osteoporosis, benign prostatic hyperplasia And can also be used in hyperactive bladders. In addition, the compounds can be used to treat recurrent stenosis formation in fibrotic diseases such as, but not limited to, pulmonary fibrosis, renal fibrosis, liver fibrosis, progressive sclerosis and Crohn's disease (Goldstein et sl. , J. Biol. Chem. 259 (14), 1984, 9263-8; Ricpero et al., J. Biol. Chem. 275 (17), 2000, 12475-80; J. Biol. Chem. 15, 2005, 14378-14384).

  They are asthma, atopic or non-atopic asthma, occupational asthma, exercise-induced bronchoconstriction, bronchitis, aluminum lung, charcoal, asbestosis, asbestosis, tendon loss, iron loss , Silicosis, pneumoconiosis including tabaccosis and pneumonia, chronic obstructive pulmonary disease including emphysema, adult respiratory distress syndrome, pneumonia, allergic rhinitis, vasculopathy, and pleurisy It is also effective for other obstacles.

  These compounds according to the present invention may be used for autoinflammatory diseases such as familial Mediterranean fever (FMF), tumor-necrosis factor receptor associated periodic syndrome (TRAPS), neonatal-onset multisystem inflammatory Diseases (neonal onset multisystem indissemination disease) (NOMID), familial cold autoinflammatory syndrome (FCAS), including familial cold urticaria (FCU), pyogenic arthritis nematode pulmonary acne pulmonary acne ) It can also be used to treat syndromes and Maccle-Wells disease.

Pain Within the present application, the term “pain” preferably refers to centrally and peripherally mediated pain, vascular pain, visceral pain, inflammatory mediated pain, neuralgia , Related pain, nociceptive pain , reflex pain, psychosomatic pain, acute injury, trauma or acute pain as caused by bone, muscle, tissue, soft tissue, organ surgery, insect Including, but not limited to, pain after biting, seizure-postpain syndrome, postoperative pain, pain associated with progressive disease, and chronic pain (see Calixto et al., Br. J. Pharmacol. 143, 2004, 803-818; Chen et al., Expert Opin. Ther. ets 11 (1), 2007, 21-35; Porreca et al., J. Pharmacol. Exp. Ther. 318 (1), 2006, 195-205; Ferreira et al., J. Neurosc. (9), 2005, 2405-12; Conley et al., Eur. J. Pharm. 527 (1-3), 2005, 44-51 ;, Levy and Zochodne, Pain 86 (3), 2000. 265-71; Yamaguchi-Sase et al., Inflamm.Res. 52 (4), 2003, 164-9).

The compounds according to the invention are preferably inflammatory pain of various origins such as rheumatoid arthritis or gout, visceral pain such as pancreatitis, interstitial cystitis, kidney or gallbladder pain, postherpetic neuralgia, etc. Neuropathic pain, complex local pain syndrome, hallucinatory pain, root avulsion, trigeminal neuralgia, painful traumatic mononeuropathy, polyneuropathy, vulvar pain, possibly any Central pain syndrome caused by some damage in the peripheral and / or central nervous system, postoperative pain syndromes such as post-mastectomy syndrome, bone and joint pain, repetitive motion pain , Toothache, cancer pain Pain including, but not limited to, fascial pain such as muscle damage and fibromyalgia, perioperative pain such as from systemic surgery, chronic pain, pain associated with dysmenorrhea and angina It is also effective in the treatment and / or prevention. In addition, these compounds are preferably effective against back pain, headache, cluster headache, migraine including prophylactic and acute use, hyperalgesia and fever. Furthermore, the compounds of the present invention are useful as analgesics for use during general and monitored anesthesia.

Diagnosis and Further Uses The present invention also provides a method for localizing or detecting B1R in a tissue, preferably a tissue section, wherein the method comprises a tissue sample presumed to contain B1R, Contacting a detectably labeled compound according to the invention under conditions allowing the binding of the compound to B1R and detecting the bound compound. Such methods and their respective conditions are known to those skilled in the art and include, for example, the radioligand binding assay disclosed in Example 24.

  The present invention also provides a method of inhibiting the binding of DAKD or other B1R ligand to B1R, the method comprising a solution containing a B1R antagonist compound disclosed herein, and a cell expressing B1R; Contacting under sufficient conditions and in a sufficient amount to detect detectable binding of DAKD or other substance to B1R. Such methods and their respective conditions are known to those skilled in the art and include, for example, the calcium kinetic assay disclosed in Example 25.

  It is also within the scope of the present invention to use a compound according to the present invention as or for the manufacture of a diagnostic agent, wherein such diagnostic agent is for therapeutic purposes as disclosed herein. It is for diagnosis of diseases and conditions that the compounds of the present invention can face.

  For various applications, the compounds of the invention can be combined with isotopes, fluorescent or luminescent markers, antibodies or antibody fragments, nanobodies, aptamers, peptides, etc., any other enzyme or enzyme substrate such as It can be labeled with any affinity label. These labeled compounds of the invention can be used in vivo, in vitro, in vitro and in situ, for example in tissue sections, to characterize BK receptors in living patients or other materials. It is useful for mapping the position of the BK receptor via radiography, and as a radio tracer in positron emission tomography (PET) imaging, single photon emission computer linked tomography (SPECT), and the like. Labeled compounds according to the invention can be used in therapeutic, diagnostic and other applications, such as in vivo and in vitro research tools, particularly in the applications disclosed herein.

  The compounds of formula (I) according to the present invention have improved properties compared to the B1R antagonists known in the state of the art, in particular improved selectivity, lower toxicity, lower drug-drug interactions, especially with regard to oral administration Bioavailability, improved metabolic stability, improved stability in microsomal degradation assays and improved solubility.

The invention will now be further illustrated by the following examples, from which further features, aspects and advantages of the invention can be understood.

EXAMPLES Abbreviations used in the following examples are as follows:
amu is an atomic mass unit.
ACN is acetonitrile.
AgNO ₃ is silver nitrate.
B1R is a B1 receptor.
BK is bradykinin.
BSA is bovine serum albumin.
conc. Is dark.
DAKD is des-Arg-10-caridine.
DCM is dichloromethane.
DIPEA is diisopropylethylamine.
DMAP is 4-N, N-dimethylaminopyridine.
DMEM is Dulbecco's modified Eagle medium.
DMF is N, N-dimethylformamide.
DMSO is dimethyl sulfoxide.
EA is ethyl acetate.
FBS is fetal bovine serum.
FCS is fetal calf serum.
h is time.
HATU is 2- (1H-7-azabenzotriazol-1-yl) -1,1,3,3-tetramethyluranium hexafluorophosphate.
HCl is hydrochloric acid.
HEK is a human embryonic kidney.
HEPES is 4-2-hydroxyethyl-1-piperazine ethanesulfonic acid.
HOBt is N-hydroxybenzotriazole.
HPLC is high performance liquid chromatography.
K ₂ CO ₃ is potassium carbonate.
IL is an interleukin.
LiOH is lithium hydroxide.
NaCl is sodium chloride.
Na ₂ CO ₃ is sodium carbonate.
Na ₂ S ₂ O ₃ is sodium thiosulfate.
NaOH is sodium hydroxide.
NaHCO ₃ is sodium bicarbonate.
NH ₄ Cl is ammonium chloride.
NH ₄ OH is ammonia.
Pd (OAc) ₂ is palladium (II) acetate.
PBS is phosphate-buffered saline.
PIPES is piperazine-N, N′-bis (2-ethanesulfonic acid).
RP is in reverse phase.
RT is room temperature.
sat. Is saturated.
tBu is tert. Butyl.
TFA is trifluoroacetate or trifluoroacetic acid.
THF is tetrahydrofuran.
TPP is Techno Plastic Products AG.
WSC is (3-dimethylamino-propyl) -ethyl-carbodiimide hydrochloride.

Materials and Methods Materials and methods and general methods are further illustrated by the following examples:
solvent:
The solvent was used in defined quality and without further purification. Acetonitrile (gradient grade, JT Baker); dichloromethane (for synthesis, Merck Eurolab); diethyl ether (for synthesis, Merck Eurolab); N, N-dimethylformamide (LAB, Merck Eurolab); dioxane (For synthesis, Aldrich); methanol (for synthesis, Merck Eurolab).

water:
Milli-Q Plus, Millipore, deionized water.

Chemicals:
The chemicals were synthesized according to or in analogy to literature methods, or Advanced ChemTech (Bamberg, Deutschland), Sigma-Aldrich-Fluka (Deisenhofen, Germany), Bachem (Heidelberg, Germany. T. T. Baker (Phillipsburg, USA), Lancaster (Muehlheim / Main, Germany), Merck Euroc (Darmstadt, Germany), Neosystem (Strassburg, France), NovaBem (Mastheim, Germany, Germany). (Geel, Belgium, sales company Fisher Scientific GmbH, Schwerte, Germany), Peptech (Cambridge, MA, USA), Syntechtech (Albany, OR, USA), Pharmacore (HighNC USA) A) and Anaspec (San Jose, CA, USA) or other companies and used in the specified quality without further purification.

  Concentrations are given as volume percent unless stated otherwise.

RP-HPLC-MS analysis:
For analytical chromatography, a Hewlett Packard 1100-system (degasser G1322A, quaternary pump G1311A, automatic sample changer G1313A, column heater G1316A, variable UV detector G1314A) was used with ESI-MS (Finigan LCQ ion trap). Mass spectrometer). The system was controlled by “navigator version 1.1 sp1” software (finnigan). Helium was used as the impact gas in the ion trap. For chromatographic separation, in all chromatograms 30 ° C. and linear gradient (5-95% B over 25 minutes, linear, A: 0.05% TFA in water and B: CH ₃ CN RP-18-column (Vydac 218 TP5215, 2.1 × 150 mm, 5 μm, C18, 300A with precolumn) (Merck)) at a flow rate of 0.3 ml / min using 0.05% TFA). UV detection was performed at λ = 220 nm. The retention time (R _t ) is shown in decimal (eg, 1.9 minutes = 1 minute 54 seconds), which refers to detection in a mass spectrometer. The dead time between injection and UV detection (HPLC) was 1.65 minutes, and the dead time between UV detection and mass detection was 0.21 minutes. The accuracy of the mass spectrometer was about ± 0.5 amu.

With a 5 μl injection, using a 95: 5 to 5:95 linear gradient (A: 0.05% TFA in water and B: 0.05% TFA in ACN) in 9.5 minutes, HPLC / MS analysis was performed. The RP column was from Phenomenex (Type Luna
C-18, 3 μm, 50 × 2.00 mm, flow rate 0.3 ml, HPLC at room temperature); mass spectrometer: ThermoFinnigan Advantage and / or LCQ Classic (both ion traps), ESI ionization, helium works as bombardment gas in ion trap It was. Excalibur version 1.3 and / or 1.2 was used as software. The retention time (R _t ) is shown in decimal (for example, 1.9 minutes = 1 minute 54 seconds).

Preparative HPLC :
Preparative HPLC separation was performed using a Vydac R18-RP column with the following gradient solvents: 0.05% TFA in H ₂ O and B: 0.05% TFA in CH ₃ CN.

  The compound was named AutoNom version 2.2 (Beilstein Information system Copyright 1988-1998, Beilstein Institute for Litter Organic Chemistry Chemistry Chem.).

Compound production:
The compounds of the present invention can be prepared in a number of ways well known to those skilled in the art of organic synthesis. The compounds of the present invention can be synthesized using the methods described below, along with synthetic methods known in the art of synthetic organic chemistry or variations thereof recognized by those skilled in the art. Preferred methods include, but are not limited to, the methods described below. Each of the references cited below becomes the content of this specification by quoting.

  The following examples show specific examples for the preparation of compounds of formula (I). Unless otherwise noted, all starting materials and reagents are of standard commercial grade and are used without further purification or are readily prepared from such materials by routine methods . Those skilled in the art of organic synthesis will recognize that the reaction conditions including the starting materials and additional steps used for the preparation of the compounds encompassed by the present invention can be varied.

(R) -pyrimidine-5-carboxylic acid (1- {1- [4- (2,5-cis / trans-dimethyl-pyrrolidine-1-carbonyl) -phenyl] -ethylcarbamoyl} -cyclopropyl) -amide ( Synthesis of compound 1)

Method 1:
A. (R)-[1- (4-Bromo-phenyl) -ethyl] -carbamic acid tert-butyl ester (R) -1- (4-Bromo-phenyl) -ethylamine hydrochloride (2.50 g, 10.5)
7 mmol) was added in portions to a stirred solution of di-tert-butyl dicarbonate (2.77 g, 12.68 mmol) and triethylamine (3.24 mL, 23.25 mmol) in ACN (40 mL). . After stirring at room temperature for 18 hours, the solvent was removed in vacuo and the residue was purified by flash chromatography on silica gel (eluting with n-hexane / EA 4: 1) to give the title compound. MS (m / z): 230.0 [M + H ⁺ -OtBu]

B. (R) -4- (1-tert-Butoxycarbonylamino-ethyl) -benzoic acid methyl ester (R)-[1- (4-Bromo-phenyl) -ethyl] -carbamic acid in anhydrous DMSO (25 mL) tert -Butyl ester (3.18 g, 10.58 mmol), 1,4-bis- (diphenylphosphino) butane (0.32 g, 0.74 mmol) and Pd (OAc) ₂ (0.16 g, 0.74) To a stirred suspension of (mmol) was added triethylamine (8.85 mL, 63.46 mmol) and anhydrous methanol (10 mL) under a nitrogen atmosphere. After stirring at 70 ° C. for 48 hours under an atmosphere of carbon monoxide, the mixture was concentrated in vacuo and the residue was partitioned between EA (200 mL) and saturated NaHCO ₃ solution (200 mL). After extracting the aqueous layer with EA (2 × 200 mL), the combined organic layers were dried over Na ₂ SO ₄ , filtered and concentrated in vacuo. The residue was purified by flash chromatography on silica gel (eluting with n-hexane / EA 19: 1) to give the title compound. MS (m / z): 208.2 [M + H ^<+ > ^- OtBu]

C. (R) -4- (1-tert-butoxycarbonylamino-ethyl) -benzoic acid (R) -4- (1-tert-butoxycarbonylamino-ethyl) -benzoic acid methyl ester (1) in dioxane (25 mL) .16 g, 4.16 mmol) and a mixture of LiOH (1M solution in H ₂ O, 25 mL) was stirred at room temperature for 3 h. The mixture was concentrated in vacuo and EA (150 mL) was added. The pH of the aqueous layer was adjusted to 4 by addition of 1N HCl solution and the aqueous layer was extracted with EA (2 × 150 mL). The combined organic layers were dried over Na ₂ SO ₄ , filtered and concentrated in vacuo. The resulting title compound was used in the next reaction without purification. MS (m / z): 209.9 [M + H ⁺ -OtBu]

D. (R)-{1- [4- (2,5-cis / trans-dimethyl-pyrrolidine-1-carbonyl) -phenyl] -ethyl} -carbamic acid tert-butyl ester in DCM / DMF 9: 1 (10 mL) (R) -4- (1-tert-butoxycarbonylamino-ethyl) -benzoic acid (0.83 g, 3.14 mmol), WSC (0.72 g, 3.76 mmol), HOBt (0.51 g, To a stirred solution of 3.76 mmol) and triethylamine (2.19 mL, 15.68 mmol) was added 2,5-cis / trans-dimethyl-pyrrolidine (0.76 mL, 6.27 mmol). After stirring at room temperature overnight, the solvent was removed in vacuo and the residue was purified by flash chromatography on silica gel (eluting with n-hexane / EA 4: 1) to give the title compound. MS (m / z): 347.0 [M + H ^<+ >]

E. (R)-[4- (1-Amino-ethyl) -phenyl]-(2,5-cis / trans-dimethyl-pyrrolidin-1-yl) -methanone (R)-{1- in DCM (45 mL) [4- (2,5-cis / trans-dimethyl-pyrrolidine-1-carbonyl) -phenyl] -ethyl} -carbamic acid tert-butyl ester (0.56 g, 1.62 mmol) and a solution of TFA (5 mL) Was stirred at room temperature for 4 hours. The reaction mixture was concentrated in vacuo and the residue was partitioned between DCM (50 mL) and 1N NaOH solution (50 mL). After extraction of the aqueous layer with DCM (2 × 50 mL), the combined organic layers were dried over Na ₂ SO ₄ , filtered and concentrated in vacuo to give the title compound. MS (m / z): 247.2 [M + H ^<+ >]

F. (R)-(1- {1- [4- (2,5-cis / trans-dimethyl-pyrrolidine-1-carbonyl) -phenyl] -ethylcarbamoyl} -cyclopropyl) -carbamic acid tert-butyl ester DMF ( 1-tert-butoxycarbonylamino-cyclopropanecarboxylic acid (9.8 mg, 0.049 mmol), WSC (9.3 mg, 0.049 mmol), HOBt (6.6 mg, 0.049 mmol) in 1 mL) And N-methylmorpholine (22.3 μL, 0.203 mmol) in (R)-[4- (1-amino-ethyl) -phenyl]-(2,5-cis / trans-dimethyl). -Pyrrolidin-1-yl) -methanone (10.0 mg, 0.041 mmol) was added. After stirring at room temperature overnight, the solvent was removed in vacuo and the residue was purified by reverse phase HPLC using a gradient of ACN in water containing 0.1% TFA to give the title compound as a TFA salt. MS (m / z): 429.9 [M + H ⁺ ]

G. (R) -1-Amino-cyclopropanecarboxylic acid {1- [4- (2,5-cis / trans-dimethyl-pyrrolidine-1-carbonyl) -phenyl] -ethyl} -amide in DCM (900 μL) R)-(1- {1- [4- (2,5-cis / trans-dimethyl-pyrrolidine-1-carbonyl) -phenyl] -ethylcarbamoyl} -cyclopropyl) -carbamic acid tert-butyl ester (8. A solution of 0 mg, 0.019 mmol) and TFA (100 μL) was stirred at room temperature for 4 hours. The reaction mixture was concentrated in vacuo to give the title compound as a TFA salt. MS (m / z): 330.0 [M + H ^<+ >]

H. (R) -pyrimidine-5-carboxylic acid (1- {1- [4- (2,5-cis / trans-dimethyl-pyrrolidine-1-carbonyl) -phenyl] -ethylcarbamoyl} -cyclopropyl) -amide ( 1- {1- [4- (2,5-cis / trans-dimethyl-pyrrolidine-1-carbonyl) -phenyl] -ethylcarbamoyl} -cyclopropyl) -carbamic acid tert-butyl ester according to the synthesis of (R) -1-Amino-cyclopropanecarboxylic acid {1- [4- (2,5-cis / trans-dimethyl-pyrrolidine-1-carbonyl) -phenyl] -ethyl} -amide (10.0 mg, 0.030 mmol) Was reacted with pyrimidine-5-carboxylic acid (4.5 mg, 0.036 mmol) to give the title compound as a TFA salt. MS (m / z): 436.1 [M + H ^<+ >]

Method 2:
Reaction AD was carried out based on the literature (Dickins et al., Chem. Eur. J. 5, 1999, 1095-1105).

A. (R) -N- [1- (4-Bromo-phenyl) -ethyl] -acetamide (R) -1- (4-Bromo-phenyl) -ethylamine (5.00 g, 25. 25) in diethyl ether (750 mL). To a stirred solution of 0 mmol) and triethylamine (4.53 mL, 32.5 mmol) was added acetyl chloride (2.13 mL, 30.0 mmol) dropwise at −15 ° C. The reaction mixture was allowed to warm to room temperature over 2 hours and then poured into 500 mL of water. The organic layer was washed with 0.1N HCl solution (1 × 500 mL) and water (1 × 500 mL), dried over K ₂ CO ₃ , filtered and concentrated in vacuo to give the title compound. MS (m / z): 243.8 [M + H ^<+ >]

B. (R) -N- [1- (4-Cyano-phenyl) -ethyl] -acetamide (R) -N- [1- (4-Bromo-phenyl) -ethyl] -acetamide in anhydrous DMF (30 mL) To a stirred solution of 3.47 g, 14.3 mmol) was added copper (I) cyanide (2.70 g, 30.1 mmol) under an atmosphere of nitrogen. After stirring at 180 ° C. overnight, the reaction mixture was concentrated in vacuo and the residue was carefully poured into 6N HCl solution (60 mL). The aqueous layer was extracted with DCM (3 × 100 mL). The organic layer was washed with water (1 × 100 mL), dried over Na ₂ SO ₄ , filtered and concentrated in vacuo to give the title compound. MS (m / z): 188.9 [M + H ^<+ >]

C. (R) -4- (1-Amino-ethyl) -benzoic acid (R) -N- [1- (4-cyano-phenyl) -ethyl] -acetamide (1.67 g, in 6N HCl solution (27 mL). 8.87 mmol) was stirred at 120 ° C. overnight. The solvent was removed in vacuo and the solid was washed with diethyl ether, filtered and dried to give the title compound as the HCl salt. MS (m / z): 165.9 [M + H ^<+ >]

D. (R) -4- (1-Amino-ethyl) -benzoic acid methyl ester (R) -4- (1-amino-ethyl) -benzoic acid (1.79 g, 8.87 mmol) in methanol (25 mL) To the stirred solution of was added 3 drops of concentrated HCl. After the mixture was stirred at 85 ° C. for 3 h, the solvent was removed in vacuo to give the title compound as the HCl salt. MS (m / z): 179.8 [M + H ^<+ >]

E. (R) -4- {1-[(1-Amino-cyclopropanecarbonyl) -amino] -ethyl} -benzoic acid methyl ester (R)-{1- [4- (2,5-cis / trans-dimethyl) -Pyrrolidine-1-carbonyl) -phenyl] -ethyl} -carbamic acid tert-butyl ester according to the synthesis of (R) -4- (1-amino-ethyl) -benzoic acid methyl ester (420 mg, 2.34 mmol) Is reacted with 1-tert-butoxycarbonylamino-cyclopropanecarboxylic acid (560 mg, 2.81 mmol) to give (R) -4- {1-[(1-tert-butoxycarbonylamino-cyclopropanecarbonyl) -amino. ] -Ethyl} -benzoic acid methyl ester. According to the synthesis of (R) -1-amino-cyclopropanecarboxylic acid {1- [4- (2,5-cis / trans-dimethyl-pyrrolidine-1-carbonyl) -phenyl] -ethyl} -amide, tert-butyl The ester (103 mg, 0.284 mmol) was treated with TFA to give the title compound as a TFA salt. MS (m / z): 262.9 [M + H ^<+ >]

F. (R) -4- [1-({1-[(pyrimidine-5-carbonyl) -amino] -cyclopropanecarbonyl} -amino) -ethyl] -benzoic acid methyl ester pyrimidine-5 in DMF (2 mL) To a stirred solution of carboxylic acid (42.3 mg, 0.341 mmol), HATU (129.6 mg, 0.341 mmol) and DIPEA (247.3 μL, 1.420 mmol) was added (R) -4- { 1-[(1-Amino-cyclopropanecarbonyl) -amino] -ethyl} -benzoic acid methyl ester (74.5 mg, 0.284 mmol) was added. After stirring overnight, the reaction mixture was concentrated in vacuo and the residue was partitioned between EA (30 mL) and saturated NaHCO ₃ solution (50 mL). After extracting the aqueous layer with EA (2 × 30 mL), the combined organic layers were dried over Na ₂ SO ₄ , filtered, and concentrated in vacuo to give the title compound without further purification. Used for reaction. MS (m / z): 368.9 [M + H ^<+ >]

G. (R) -pyrimidine-5-carboxylic acid (1- {1- [4- (2,5-cis / trans-dimethyl-pyrrolidine-1-carbonyl) -phenyl] -ethylcarbamoyl} -cyclopropyl) -amide ( According to the synthesis of R) -4- (1-tert-butoxycarbonylamino-ethyl) -benzoic acid, (R) -4- [1-({1-[(pyrimidine-5-carbonyl) -amino] -cyclopropane Carbonyl} -amino) -ethyl] -benzoic acid methyl ester (192 mg, 0.521 mmol) is reacted with LiOH to give (R) -4- [1-({1-[(pyrimidine-5-carbonyl) -amino]. ] -Cyclopropanecarbonyl} -amino) -ethyl] -benzoic acid. Followed by (R)-(1- {1- [4- (2,5-cis / trans-dimethyl-pyrrolidine-1-carbonyl) -phenyl] -ethylcarbamoyl} -cyclopropyl) -carbamic acid tert-butyl ester Reaction with 2,5-cis / trans-dimethyl-pyrrolidine following synthesis gave the title compound as the TFA salt. MS (m / z): 436.1 [M + H ^<+ >]

(R) -N- {1- [1- (4-Diisopropylcarbamoyl-phenyl) -ethylcarbamoyl] -cyclopropyl} -5-trifluoromethyl-nicotinamide (Synthesis of Compound 43)

A. (R) -4- {1-[(1-tert-butoxycarbonylamino-cyclopropanecarbonyl) -amino] -ethyl} -benzoic acid methyl ester (R) -4- [1-({1-[(pyrimidine According to the synthesis of -5-carbonyl) -amino] -cyclopropanecarbonyl} -amino) -ethyl] -benzoic acid methyl ester, (R) -4- (1-amino-ethyl) -benzoic acid methyl ester (1.03 g , 4.77 mmol) was reacted with 1-tert-butoxycarbonylamino-cyclopropanecarboxylic acid (0.80 g, 3.97 mmol). The crude compound was purified by flash chromatography on silica gel (eluting with n-hexane / EA 3: 7) to give the title compound. MS (m / z): 362.7 [M + H ⁺ ]

B. (R) -4- {1-[(1-Amino-cyclopropanecarbonyl) -amino] -ethyl} -N, N-diisopropyl-benzamide (R) -4- (1-tert-butoxycarbonylamino-ethyl) -According to the synthesis of benzoic acid, (R) -4- {1-[(1-tert-butoxycarbonylamino-cyclopropanecarbonyl) -amino] -ethyl} -benzoic acid methyl ester (423 mg, 1.17 mmol) is obtained. Reaction with LiOH gave (R) -4- {1-[(1-tert-butoxycarbonylamino-cyclopropanecarbonyl) -amino] -ethyl} -benzoic acid. (R) -4- [1-({1-[(Pyrimidine-5-carbonyl) -amino] -cyclopropanecarbonyl} -amino) -ethyl] -benzoic acid According to the synthesis of methyl ester, benzoic acid (59.6 mg , 0.171 mmol) was reacted with diisopropyl-amine (71.9 μL, 0.513 mmol). Purification of the crude compound by reverse phase HPLC using a gradient of ACN in water containing 0.1% TFA followed by (R) -1-amino-cyclopropanecarboxylic acid {1- [4- (2,5-cis / trans Reaction with TFA following the synthesis of -dimethyl-pyrrolidine-1-carbonyl) -phenyl] -ethyl} -amide gave the title compound as the TFA salt. MS (m / z): 332.0 [M + H ^<+ >]

C. (R) -N- {1- [1- (4-Diisopropylcarbamoyl-phenyl) -ethylcarbamoyl] -cyclopropyl} -5-trifluoromethyl-nicotinamide (R) -4- (1-tert-butoxycarbonyl According to the synthesis of amino-ethyl) -benzoic acid methyl ester, 3-bromo-5-trifluoromethyl-pyridine (1.00 g, 4.42 mmol) is reacted with carbon monoxide to give 5-trifluoromethyl-nicotinic acid. The methyl ester was given. Hydrolysis of the methyl ester following the synthesis of (R) -4- (1-tert-butoxycarbonylamino-ethyl) -benzoic acid followed by (R) -4- [1-({1-[(pyrimidine-5-carbonyl ) -Amino] -cyclopropanecarbonyl} -amino) -ethyl] -benzoic acid According to the synthesis of methyl ester, benzoic acid (4.80 mg, 0.025 mmol) of (R) -4- {1-[(1-amino Reaction with -cyclopropanecarbonyl) -amino] -ethyl} -N, N-diisopropyl-benzamide (4.20 mg, 0.013 mmol) gave the title compound as a TFA salt (containing 0.1% TFA). Purification of the crude compound by reverse phase HPLC using a gradient of ACN in water). MS (m / z): 505.1 [M + H ^<+ >]

(R) -N- {1- [1- (4-Diisopropylcarbamoyl-phenyl) -ethylcarbamoyl] -cyclopropyl} -3-fluoro-5-trifluoromethyl-benzamide (Synthesis of Compound 44)

According to the synthesis of (R) -4- [1-({1-[(pyrimidine-5-carbonyl) -amino] -cyclopropanecarbonyl} -amino) -ethyl] -benzoic acid methyl ester, (R) -4- {1-[(1-Amino-cyclopropanecarbonyl) -amino] -ethyl} -N, N-diisopropyl-benzamide (4.2 mg, 0.013 mmol) was converted to 3-fluoro-5-trifluoromethyl-benzoic acid. (5.3 mg, 0.025 mmol). Purification of the crude compound by reverse phase HPLC using a gradient of ACN in water containing 0.1% TFA gave the title compound as a TFA salt. MS (m / z): 522.1 [M + H ^<+ >]

(R) -pyrimidine-5-carboxylic acid (1- {1- [4-((2R, 5S) -2,5-dimethyl-pyrrolidine-1-carbonyl) -phenyl] -ethylcarbamoyl} -cyclopropyl)- Synthesis of amide (compound 45)

  Reaction AC was performed based on the literature (Katritzky et al., J. Org. Chem. 64, 1999, 1979-1985).

A. (3S, 5R, 7aR) -5- (benzotriazol-1-yl) -3-phenyl [2,1-b] oxazolopyrrolidine 2,5-dimethoxytetrahydrofuran (2.94 mL, 22.7 mmol) and 0 A mixture of .1N HCl (90 mL) was refluxed for 1 hour and then cooled to room temperature. To the cooled mixture was added benzotriazole (2.70 g, 22.7 mmol) and (S)-(+)-2-amino-2-phenyl-ethanol (3.11 g, 22. 7 mmol) was added and it was then stirred overnight. The reaction mixture was washed with 2N NaOH (3 × 70 mL) and water (2 × 70 mL). The organic layer was dried over Na ₂ SO ₄ and filtered. After removal of the solvent, the residue was recrystallized from EA to give the title compound. ¹ H NMR δ 2.20-2.26 (m, 1H), 2.55-2.57 (m, 3H), 3.61-3.68 (m, 1H), 4.44-4.45 (M, 2H), 5.16-5.17 (m, 1H), 5.90-5.94 (m, 1H), 7.03-7.08 (m, 5H), 7.21-7 .26 (m, 2H), 7.44-7.46 (d, J = 7 Hz, 1H), 7.90-7.93 (d, J = 7 Hz, 1H)

B. (2S)-(2- (2R, 5S) -2,5-Dimethyltetrahydro-1H-1-pyrrolyl-2-phenylethane-1-ol (3S, 5R, 7aR) -5 in dry THF (33 mL) To a stirred solution of-(benzotriazol-1-yl) -3-phenyl [2,1-b] oxazolopyrrolidine (500 mg, 1.63 mmol) was added a solution of methylmagnesium bromide in diethyl ether under nitrogen. 3M, 2.20 mL, 6.53 mmol) was added dropwise at 0 ° C. After stirring overnight at room temperature, the reaction mixture was washed with 2N NaOH (3 × 40 mL) and water (2 × 40 mL) .The organic layer was Na ₂ SO _4. Dried over, filtered and concentrated in vacuo The residue was flash chromatographed on silica gel (eluted with n-hexane / EA 19: 1) More purified,. MS gave the title compound (m / z): 220.0 [ M + H +]

C. (2R, 5S) -2,5-dimethyltetrahydro-1H-1-pyrrolidinium chloride (2S)-(2- (2R, 5S) -2,5-dimethyltetrahydro-1H-1-pyrrolyl-2-phenyl ethane-1-ol (100 mg, 0.46 mmol) Thales H-Cube containing cartridge 10% palladium on carbon as a. catalyst dissolved in methanol ^{(9mL) (R)} flow Hydrogenics transilluminator (flow hydrogenator ) (Thales Nanotechnology, H-1031 Budapest, Zahony utca 7 (Graphissoft Park), Hungary. Website: http://www.thalesano.com). Hydrogenation was carried out at room temperature under pressure of bar The product solution was treated with 20 drops of concentrated HCl with stirring at room temperature for 30 minutes The solvent was evaporated in vacuo and the resulting product was diluted with diethyl Washing with ether gave the title compound as the HCl salt, MS (m / z): 100.00 [M + H ⁺ ].

D. (R) -pyrimidine-5-carboxylic acid (1- {1- [4-((2R, 5S) -2,5-dimethyl-pyrrolidine-1-carbonyl) -phenyl] -ethylcarbamoyl} -cyclopropyl)- Amide (R) -4- [1-({1-[(pyrimidine-5-carbonyl) -amino] -cyclopropanecarbonyl} -amino) -ethyl] -benzoic acid according to the synthesis of methyl ester (R) -4 -{1-[(1-tert-Butoxycarbonylamino-cyclopropanecarbonyl) -amino] -ethyl} -benzoic acid (30 mg, 0.086 mmol) was added to (2R, 5S) -2,5-dimethyltetrahydro-1H Reaction with -1-pyrrolidinium chloride (23 mg, 0.172 mmol) gives (R)-(1- {1- [4-((2R, 5S) -2,5-dimethyl). - pyrrolidine-1-carbonyl) - phenyl] - ethylcarbamoyl} - Shikupuropiru) - give carbamic acid tert- butyl ester. (R) -1-Amino-cyclopropanecarboxylic acid {1- [4- (2,5-cis / trans-dimethyl-pyrrolidine-1-carbonyl) -phenyl] -ethyl} -amide with TFA following synthesis The reaction gives (R) -1-amino-cyclopropanecarboxylic acid {1- [4-((2R, 5S) -2,5-dimethyl-pyrrolidine-1-carbonyl) -phenyl] -ethyl} -amide. It was. Pyrimidine-5-carboxylic acid following the synthesis of (R) -4- [1-({1-[(pyrimidine-5-carbonyl) -amino] -cyclopropanecarbonyl} -amino) -ethyl] -benzoic acid methyl ester And purification of the crude product by reverse phase HPLC using a gradient of ACN in water containing 0.1% TFA gave the title compound as a TFA salt. MS (m / z): 436.1 [M + H ^<+ >]

(R) -N- (1- {1- [4- (2,5-cis / trans-dimethyl-2,5-dihydro-pyrrol-1-carbonyl) -phenyl] -ethylcarbamoyl} -cyclopropyl)- Synthesis of 3-fluoro-5-trifluoromethyl-benzamide (compound 55)

According to the synthesis of (R) -4- [1-({1-[(pyrimidine-5-carbonyl) -amino] -cyclopropanecarbonyl} -amino) -ethyl] -benzoic acid methyl ester, (R) -4- {1-[(1-tert-Butoxycarbonylamino-cyclopropanecarbonyl) -amino] -ethyl} -benzoic acid (60 mg, 0.17 mmol) was added to 2,5-cis / trans-dimethyl-2,5-dihydro. -1H-pyrrole (33 mg, 0.34 mmol) and (R)-(1- {1- [4- (2,5-cis / trans-dimethyl-2,5-dihydro-pyrrole-1- Carbonyl) -phenyl] -ethylcarbamoyl} -cyclopropyl) -carbamic acid tert-butyl ester was obtained. (R) -1-Amino-cyclopropanecarboxylic acid {1- [4- (2,5-cis / trans-dimethyl-pyrrolidine-1-carbonyl) -phenyl] -ethyl} -amide with TFA following synthesis The reaction is (R) -1-amino-cyclopropanecarboxylic acid {1- [4- (2,5-cis / trans-dimethyl-2,5-dihydro-pyrrole-1-carbonyl) -phenyl] -ethyl} -The amide was given. The following (R) -4- [1-({1-[(pyrimidine-5-carbonyl) -amino] -cyclopropanecarbonyl} -amino) -ethyl] -benzoic acid methyl ester follows the synthesis of 3-fluoro-5- Purification of the crude compound by reverse phase HPLC using a reaction with trifluoromethyl-benzoic acid and a gradient of ACN in water containing 0.1% TFA gave the title compound as the TFA salt. MS (m / z): 518.2 [M + H ^<+ >]

(R, S) -pyrimidine-5-carboxylic acid (1- {1- [4- (2,5-cis / trans-dimethyl-pyrrolidine-1-carbonyl) -2-fluoro-phenyl] -ethylcarbamoyl}- Synthesis of cyclopropyl) -amide (compound 60)

A. 3-Fluoro-4-methyl-benzoic acid methyl ester To a stirred solution of 3-fluoro-4-methyl-benzoic acid (1.11 g, 7.20 mmol) in methanol (24 mL) was added chloro-trimethyl-silane (3 .64 mL, 28.8 mmol) was added. After stirring at room temperature overnight, the solvent was removed in vacuo and the residue was purified by flash chromatography on silica gel (eluting with n-hexane) to give the title compound. GC / MS (m / z): 168.0
Reaction B was performed based on literature (Gauuan et al., Bioorg. Med. Chem. 10, 2002, 3013-3021).

B. 3-Fluoro-4-formyl-benzoic acid methyl ester 3-Fluoro-4-methyl-benzoic acid methyl ester (500 mg, 2.97 mmol), 1-bromo-pyrrolidine-2,5 in tetrachloromethane (36 mL) After stirring a suspension of dione (1.73 g, 8.33 mmol) and benzoyl peroxide (53.1 mg, 0.47 mmol) at 80 ° C. overnight, the mixture was concentrated in vacuo to give a residue Was partitioned between EA (150 mL) and water (100 mL). The organic layer was washed with saturated NaHCO ₃ solution (1 × 100 mL), water (1 × 100 mL) and saturated NaCl solution (1 × 100 mL). The organic layer was dried over Na ₂ SO ₄ , filtered and concentrated in vacuo. A mixture of the residue, 4-bromomethyl-3-fluoro-benzoic acid methyl ester and 4-dibromomethyl-3-fluoro-benzoic acid methyl ester, was used in the subsequent step without further purification. The residue (50 mg, 0.30 mmol) was dissolved in acetone (2 mL) and water (0.4 mL) and AgNO ₃ (171 mg, 0.87 mmol) was added. The flask was covered with aluminum foil to avoid decomposition of AgNO ₃ . The mixture was stirred at room temperature overnight. After filtration of the mixture and evaporation of the solvent, the residue was partitioned between EA (30 mL) and saturated NaHCO ₃ solution (15 mL). The organic layer was washed with water (1 × 15 mL) and saturated NaCl solution (1 × 15 mL), dried over Na ₂ SO ₄ , filtered and concentrated in vacuo to give the title compound. GC / MS (m / z): 182.2

C. (R, S) -3-Fluoro-4- (1-hydroxy-ethyl) -benzoic acid methyl ester 3-Fluoro-4-formyl-benzoic acid methyl ester (450 mg, 2.47 mmol) in dry THF (10 mL). ) Was added dropwise at −80 ° C. under nitrogen at −80 ° C. with methylmagnesium bromide (3M solution in diethyl ether, 0.99 mL, 2.96 mmol). After stirring for 80 minutes, 5 mL of 2M HCl solution was added, allowing the mixture to warm to room temperature. Saturated NaCl solution was added (100 mL) and the mixture was extracted with EA (4 × 100 mL). The organic layer was dried over Na ₂ SO ₄ , filtered and concentrated in vacuo. The residue was purified by flash chromatography on silica gel (eluting with n-hexane / EA 6: 1) to give the title compound. GC / MS (m / z): 198.2

D. (R, S) -4- (1-Azido-ethyl) -3-fluoro-benzoic acid methyl ester Methanesulfonyl chloride (120 μL, 1.55 mmol) was dissolved in 500 μL DCM and then in DCM (4 mL). Of (R, S) -3-fluoro-4- (1-hydroxy-ethyl) -benzoic acid methyl ester (154 mg, 0.78 mmol), DMAP (4.7 mg, 0.04 mmol) and triethylamine (1. (08 mL, 7.77 mmol) was added dropwise at 0 ° C. to the stirred solution. After stirring at 0 ° C. for 30 minutes, 10 mL of water was added and the mixture was extracted with DCM (3 × 50 mL). The organic layer was dried over Na ₂ SO ₄ , filtered and concentrated in vacuo to give cis / trans-3-fluoro-4- (1-methanesulfonyloxy-ethyl) -benzoic acid methyl ester, which was Used in subsequent steps without further purification. The residue was stirred with sodium azide (204 mg, 4.66 mmol) in ACN (4 mL) at 60 ° C. overnight. The solvent was removed in vacuo and the residue was partitioned between DCM (50 mL) and water (50 mL). The aqueous layer was extracted with DCM (2 × 20 mL). The organic layer was dried over Na ₂ SO ₄ , filtered and concentrated in vacuo. The residue was purified by flash chromatography on silica gel (eluting with n-hexane / EA 9: 1) to give the title compound. GC / MS (m / z): 223.0

E. (R, S) -4- (1-Amino-ethyl) -3-fluoro-benzoic acid methyl ester (R, S) -4- (1-azido-ethyl) -3-fluoro-benzoic acid methyl ester (158 mg , 0.71 mmol) was dissolved in methanol (14 mL). Theles H-Cube ^(R) containing a 10% palladium on carbon cartridge as catalyst
The solution was subjected to continuous flow hydrogenation using a flow hydrogenerator. Hydrogenation was carried out at room temperature under a pressure of 1 bar. The solvent was evaporated to give the title compound. MS (m / z): 197.8 [M + H ^<+ >]

F. (R, S) -3-fluoro-4- [1-({1-[(pyrimidine-5-carbonyl) -amino] cyclopropanecarbonyl} -amino) -ethyl] -benzoic acid methyl ester (R, S) -4- (1-amino-ethyl) -3-fluoro-benzoic acid methyl ester (27.4 mg, 0.14 mmol) is reacted with 1-tert-butoxycarbonylamino-cyclopropanecarboxylic acid and then (R) Treated with TFA according to the synthesis of -4- {1-[(1-amino-cyclopropanecarbonyl) -amino] -ethyl} -benzoic acid methyl ester. Pyrimidine-5-carboxylic acid following the synthesis of (R) -4- [1-({1-[(pyrimidine-5-carbonyl) -amino] -cyclopropanecarbonyl} -amino) -ethyl] -benzoic acid methyl ester Reaction with gave the title compound. MS (m / z): 386.9 [M + H ^<+ >]

G. (R, S) -pyrimidine-5-carboxylic acid (1- {1- [4- (2,5-cis / trans-dimethyl-pyrrolidine-1-carbonyl) -2-fluoro-phenyl] -ethylcarbamoyl}- Cyclopropyl) -amide (R, S) -3-fluoro-4- [1-({1-[(pyrimidine-5-carbonyl) -amino] cyclopropanecarbonyl} -amino) -ethyl] -benzoic acid methyl ester (73.0 mg, 0.19 mmol) was reacted with LiOH according to the synthesis of (R) -4- (1-tert-butoxycarbonylamino-ethyl) -benzoic acid. Subsequent (R) -4- [1-({1-[(pyrimidine-5-carbonyl) -amino] -cyclopropanecarbonyl} -amino) -ethyl] -benzoic acid benzoic acid (13. Reaction with 0 mg, 0.04 mmol) 2,5-cis / trans-dimethyl-pyrrolidine (4.30 μL, 0.04 mmol) and reverse phase HPLC using a gradient of ACN in water containing 0.1% TFA. Purification of the crude compound by gave the title compound as a TFA salt. MS (m / z): 454.1 [M + H ⁺ ]

(R, S) -N- (1- {1- [5- (2,5-cis / trans-dimethyl-pyrrolidin-1-carbonyl) -pyridin-2-yl] -ethylcarbamoyl} -cyclopropyl)- Synthesis of 5-trifluoromethyl-nicotinamide (Compound 65)

  Reaction A was performed based on the literature (Chong et al., J. Med. Chem. 47, 2004, 5230-5234).

A. 6-Hydroxymethyl-nicotinic acid methyl ester pyridine-2,5-dicarboxylic acid dimethyl ester (1.05 g, 5.38 mmol) and calcium chloride (2.39 g, 21.21) in THF / methanol 1: 2 (33 mL). To a stirred suspension of 5 mmol) sodium borohydride (0.51 g, 13.5 mmol) was added in portions at 0 ° C. After stirring at 0 ° C. overnight, 20 mL of water was added and the solvent was removed in vacuo. The aqueous residue was extracted with DCM (3 × 30 mL) and the organic layer was dried over Na ₂ SO ₄ , filtered and concentrated in vacuo. The residue was purified by flash chromatography on silica gel (eluting with n-hexane / EA 1: 1) to give the title compound. MS (m / z): 167.9 [M + H ^<+ >]

B. 6-Formyl-nicotinic acid methyl ester To a stirred solution of 6-hydroxymethyl-nicotinic acid methyl ester (55 mg, 0.33 mmol) in DCM (17 mL) was added Dess-Martin reagent (207 mg, 0.49 mmol). It was. After stirring for 3 hours, the solvent was removed in vacuo. The residue was purified by flash chromatography on silica gel (eluting with n-hexane / EA 4: 1) to give the title compound. GCMS (m / z): 165.2

C. (R, S) -6- (1-Hydroxy-ethyl) -nicotinic acid methyl ester According to the synthesis of (R, S) -3-fluoro-4- (1-hydroxy-ethyl) -benzoic acid methyl ester, 6- Formyl-nicotinic acid methyl ester (47 mg, 0.28 mmol) was reacted with methylmagnesium bromide to give the title compound, which was used in subsequent reactions without further purification. MS (m / z): 182.0 [M + H ^<+ >]

D. (R, S) -6- (1-Amino-ethyl) -nicotinic acid methyl ester (R, S) -6- (1-hydroxy-ethyl) -nicotinic acid methyl ester (150 mg, 0.83 mmol) and methane Reaction of the sulfonyl chloride followed by reaction with sodium azide following the synthesis of (R, S) -4- (1-azido-ethyl) -3-fluoro-benzoic acid methyl ester was performed using (R, S) -6- (1 -Azido-ethyl) -nicotinic acid methyl ester was provided. Hydrogenation of the methyl ester following the synthesis of (R, S) -4- (1-amino-ethyl) -3-fluoro-benzoic acid methyl ester gave the title compound. MS (m / z): 181.0 [M + H ⁺ ]

E. (R, S)-(1- {1- [5- (2,5-cis / trans-dimethyl-pyrrolidin-1-carbonyl) -pyridin-2-yl] -ethylcarbamoyl} -cyclopropyl)
-Carbamic acid tert-butyl ester (R) -4- [1-({1-[(pyrimidine-5-carbonyl) -amino] -cyclopropanecarbonyl} -amino) -ethyl] -benzoic acid according to the synthesis of methyl ester , (R, S) -6- (1-amino-ethyl) -nicotinic acid methyl ester (116 mg, 0.64 mmol) was reacted with 1-tert-butoxycarbonylamino-cyclopropanecarboxylic acid. Purification of the crude compound by reverse phase HPLC using a gradient of ACN in water containing 0.1% TFA was obtained by (R, S) -6- {1-[(1-tert-butoxycarbonylamino-cyclopropanecarbonyl)- Amino] -ethyl} -nicotinic acid methyl ester was provided as the TFA salt. Reaction of methyl ester with LiOH followed by (R) -pyrimidine-5-carboxylic acid (1- {1- [4- (2,5-cis / trans-dimethyl-pyrrolidine-1-carbonyl) -phenyl] -ethyl Reaction with 2,5-cis / trans-dimethyl-pyrrolidine following the synthesis of carbamoyl} -cyclopropyl) -amide gave the title compound as the TFA salt. MS (m / z): 431.0 [M + H ^<+ >]

F. (R, S) -N- (1- {1- [5- (2,5-cis / trans-dimethyl-pyrrolidin-1-carbonyl) -pyridin-2-yl] -ethylcarbamoyl} -cyclopropyl)- 5-trifluoromethyl-nicotinamide (R) -1-amino-cyclopropanecarboxylic acid {1- [4- (2,5-cis / trans-dimethyl-pyrrolidine-1-carbonyl) -phenyl] -ethyl}- According to the synthesis of amide, (R, S)-(1- {1- [5- (2,5-cis / trans-dimethyl-pyrrolidin-1-carbonyl) -pyridin-2-yl] -ethylcarbamoyl} -cyclo Propyl) -carbamic acid tert-butyl ester (33.6 mg, 0.08 mmol) was treated with TFA. The following (R) -4- [1-({1-[(pyrimidine-5-carbonyl) -amino] -cyclopropanecarbonyl} -amino) -ethyl] -benzoic acid methyl ester according to the synthesis of 5-trifluoromethyl- Purification of the crude compound by reverse phase HPLC using a reaction with nicotinic acid and a gradient of ACN in water containing 0.1% TFA gave the title compound as the TFA salt. MS (m / z): 504.1 [M + H ⁺ ]

Synthesis of N- (1- (4- (2,5-dimethylpyrrolidine-1-carbonyl) phenyl) ethyl) -1- (1-hydroxycyclopropanecarboxyamide) cyclopropanecarboxyamide (Compound 7)

According to the synthesis of (1- {1- [4- (2,5-cis / trans-dimethyl-pyrrolidine-1-carbonyl) -phenyl] -ethylcarbamoyl} -cyclopropyl) -carbamic acid tert-butyl ester, (R ) -1-Amino-cyclopropanecarboxylic acid {1- [4- (2,5-cis / trans-dimethyl-pyrrolidine-1-carbonyl) -phenyl] -ethyl} -amide (10.0 mg, 0.03 mmol) ) Was reacted with 1-hydroxy-cyclopropanecarboxylic acid (2.8 mg, 0.03 mmol) to give the title compound as a TFA salt. MS (m / z): 414.1 [M + H ^<+ >]

N- {1- [4- (2,5-cis / trans-dimethyl-pyrrolidine-1-carbonyl)
Synthesis of 2-fluoro-benzylcarbamoyl] -cyclopropyl} -5-trifluoromethyl-nicotinamide (Compound 78)

A. 3-Fluoro-4-formyl-benzoic acid A solution of 3-fluoro-4-formyl-benzoic acid methyl ester (200 mg, 1.10 mmol) and LiOH (1.5 mL) in ACN (1.5 mL) at room temperature. Stir for 1 hour and then at 40 ° C. for 1 hour. The solvent was removed in vacuo and the residue was purified by reverse phase HPLC using a gradient of ACN in water containing 0.1% TFA to give the title compound as a TFA salt. GC / MS (m / z): 168.0

B. 4- (2,5-cis / trans-dimethyl-pyrrolidine-1-carbonyl) -2-fluoro-benzaldehyde 3-fluoro-4-formyl-benzoic acid (150 mg, 0.89 mmol) in DMF (3 mL), To a stirred solution of WSC (256 mg, 1.34 mmol) and HOBt (180 mg, 1.34 mmol) was added 2,5-cis / trans-dimethyl-pyrrolidine (264 mg, 2.67 mmol). After stirring at room temperature for 3 hours, the solvent was removed in vacuo and the residue was purified by reverse phase HPLC using a gradient of ACN in water containing 0.1% TFA to give the title compound as a TFA salt. MS (m / z): 250.2 [M + H ⁺ ]

C. (4-Aminomethyl-3-fluoro-phenyl)-(2,5-cis / trans-dimethyl-pyrrolidin-1-yl) -methanone 4- (2,5-cis / trans-dimethyl-pyrrolidine-1-carbonyl ) A mixture of 2-fluoro-benzaldehyde (50.0 mg, 0.20 mmol), titanium-tetra-isopropylate (570 mg, 2.00 mmol) and ammonia (7N solution in ethanol, 1 mL) was stirred at room temperature overnight. did. Sodium borohydride (23.0 mg, 0.60 mmol) was added. After stirring for 2 hours, the solvent was removed in vacuo and the residue was partitioned between EA (20 mL) and saturated NaHCO ₃ solution (20 mL). Extract the aqueous layer with EA (3 × 20 mL) and dry the organic layer over Na ₂ SO ₄ , filter and concentrate in vacuo to give the title compound, MS (m / z): 251. 1 [M + H ⁺ ]

D. {1- [4- (2,5-cis / trans-dimethyl-pyrrolidine-1-carbonyl) -2-fluoro-benzylcarbamoyl] -cyclopropyl} -carbamic acid tert-butyl ester 1- in DMF (1 mL) tert-Butoxycarbonylamino-cyclopropanecarboxylic acid (80.5 mg, 0.40 mmol), WSC (77.0 mg, 0.40 mmol), HOBt (54.0 mg, 0.40 mmol) and diisopropylamine (100 μL, 0.60 mmol) was added to (4-aminomethyl-3-fluoro-phenyl)-(2,5-cis / trans-dimethyl-pyrrolidin-1-yl) -methanone (50.1 mg, 0.20). Mmol) was added. After stirring at room temperature for 3 hours, the solvent was removed in vacuo and the residue was purified by reverse phase HPLC using a gradient of ACN in water containing 0.1% TFA to give the title compound as a TFA salt, MS (M / z): 433.9 [M + H ⁺ ]

E. 1-amino-cyclopropanecarboxylic acid 4- (2,5-cis / trans-dimethyl-pyrrolidine-1-carbonyl) -2-fluoro-benzylamide {1- [4- (2,5) in DCM (2700 μL) -Cis / trans-dimethyl-pyrrolidine-1-carbonyl) -2-fluoro-benzylcarbamoyl] -cyclopropyl} -carbamic acid tert-butyl ester (33 mg, 0.08 mmol) and TFA (300 μL) were added at room temperature. For 4 hours. The reaction mixture was concentrated in vacuo. The residue was purified by reverse phase HPLC using a gradient of ACN in water containing 0.1% TFA to give the title compound as a TFA salt. MS (m / z): 334.1 [M + H ^<+ >]

F. 1-amino-cyclopropanecarboxylic acid 4- (2,5-cis / trans-dimethyl-pyrrolidine-1-carbonyl) -2-fluoro-benzylamide {1- [4- (2,5-cis / trans-dimethyl) -Pyrrolidine-1-carbonyl) -2-fluoro-benzylcarbamoyl] -cyclopropyl} -carbamic acid tert-butyl ester according to the synthesis of 1-amino-cyclopropanecarboxylic acid 4- (2,5-cis / trans-dimethyl -Pyrrolidine-1-carbonyl) -2-fluoro-benzylamide (13 mg, 0.04 mmol) was reacted with 5-trifluoromethyl-nicotinic acid (33 mg, 0.08 mmol) to give the title compound as the TFA salt. It was. MS (m / z): 507.1 [M + H ^<+ >]

(R) -N- (1- {1- [4-((2R, 5S) -2,5-dimethyl-pyrrolidine-1-carbonyl) -2-fluoro-phenyl] -ethylcarbamoyl} -cyclopropyl)- Synthesis of 3-fluoro-5-trifluoromethyl-benzamide (compound 90)

A. 3-Fluoro-4-[(2-methyl-propane-2-sulfinylimino) -methyl] -benzoic acid methyl ester 3-Fluoro-4-formyl-benzoic acid methyl ester (529 mg, 2. 91 mmol), (S)-(−) 2-methyl-propane-2-sulfinic acid amide (352 mg, 2.91 mmol), pyridinium p-toluenesulfonate (36 mg, 0.15 mmol) and magnesium sulfate (5. 20 g, 43.7 mmol) was stirred at room temperature for 2 days. The solvent was removed in vacuo and the residue was purified by flash chromatography on silica gel (eluting with n-hexane / EA 9: 1) to give the title compound. MS (m / z): 285.8 [M + H ^<+ >]

B. (R) -3-Fluoro-4- [1- (2-methyl-propane-2-sulfinylamino) -ethyl] -benzoic acid methyl ester 3-fluoro-4-[(2-methyl) in DCM (7 mL) -Propane-2-sulfinylimino) -methyl] -benzoic acid methyl ester (386 mg, 1.35 mmol) was added to methylmagnesium chloride (22% in THF, 520 μL, 1.49) at −60 ° C. Mmol) was added dropwise. After stirring at −60 ° C. for 4 hours, saturated ammonium chloride solution (25 mL) was added. The aqueous layer was extracted with DCM (4 × 25 mL) and the organic layer was dried over Na ₂ SO ₄ , filtered and concentrated in vacuo. The residue was purified by flash chromatography on silica gel (eluting with n-hexane / EA 1: 1) to give the title compound. MS (m / z): 301.8 [M + H ⁺ ]

C. (R) -4- (1-Amino-ethyl) -3-fluoro-benzoic acid methyl ester (R) -3-fluoro-4- [1- (2-methyl-propane-2-sulfinylamino) -ethyl] A solution of benzoic acid methyl ester (61 mg, 0.21 mmol) and HCl (4N solution in dioxane, 320 μL, 1.28 mmol) was stirred at room temperature for 30 min. The solvent was removed in vacuo and the residue was recrystallized from diethyl ether to give the title compound as the HCl salt. MS (m / z): 197.8 [M + H ^<+ >]

D. (R) -3-Fluoro-4- (1-{[1- (3-fluoro-5-trifluoromethyl-benzoylamino) -cyclopropanecarbonyl] -amino} -ethyl) -benzoic acid methyl ester (R) 4- (1-Amino-ethyl) -3-fluoro-benzoic acid methyl ester (35 mg, 0.15 mmol) is reacted with 1-tert-butoxycarbonylamino-cyclopropanecarboxylic acid and then (R) -4 Treated with TFA according to the synthesis of-{1-[(1-amino-cyclopropanecarbonyl) -amino] -ethyl} -benzoic acid methyl ester. The following (R) -4- [1-({1-[(pyrimidine-5-carbonyl) -amino] -cyclopropanecarbonyl} -amino) -ethyl] -benzoic acid methyl ester follows the synthesis of 3-fluoro-5- Reaction with trifluoromethyl-benzoic acid gave the title compound. MS (m / z): 470.8 [M + H ⁺ ]

E. (R) -N- (1- {1- [4-((2R, 5S) -2,5-dimethyl-pyrrolidine-1-carbonyl) -2-fluoro-phenyl] -ethylcarbamoyl} -cyclopropyl)- 3-Fluoro-5-trifluoromethyl-benzamide According to the synthesis of (R) -4- (1-tert-butoxycarbonylamino-ethyl) -benzoic acid, (R) -3-fluoro-4- (1-{[ 1- (3-Fluoro-5-trifluoromethyl-benzoylamino) -cyclopropanecarbonyl] -amino} -ethyl) -benzoic acid methyl ester (98 mg, 0.21 mmol) was reacted with LiOH. Benzoic acid following the synthesis of (R) -4- [1-({1-[(pyrimidine-5-carbonyl) -amino] -cyclopropanecarbonyl} -amino) -ethyl] -benzoic acid methyl ester (8.4 mg , 0.02 mmol) of (2R, 5S) -2,5-dimethyltetrahydro-1H-1-pyrrolidinium chloride (3.2 mg, 0.024 mmol) and water containing 0.1% TFA Purification of the crude compound by reverse phase HPLC using a gradient of ACN gave the title compound as the TFA salt. MS (m / z): 538.3 [M + H ⁺ ]

{6- [4- (2,5-cis / trans-dimethyl-pyrrolidin-1-carbonyl) -2-fluoro-benzylamino] -pyridin-3-yl}-(4-pyridin-4-yl-piperazine- Synthesis of 1-yl) -methanone (compound 106)

A. 6-tert-Butoxycarbonylamino-nicotinic acid methyl ester To a stirred solution of di-tert-butyl dicarbonate (1.72 g, 7.89 mmol) in tert-butanol (20 mL) was added 6-amino-nicotinic acid methyl ester. (1.00 g, 6.57 mmol) was added in portions. After stirring at room temperature for 4 days, the solvent was removed in vacuo and the residue was flash chromatographed on silica gel (n-hexane / EA).
Elution with 9: 1) to give the title compound. MS (m / z): 252.8 [M + H ^<+ >]

B. 6-tert-butoxycarbonylamino - 6-tert-butoxycarbonylamino in nicotinic acid-dioxane (40 mL) - nicotinic acid methyl ester (1.62 g, 6.40 mmol) and LiOH _{(H 2} 1M solution in O, 40 mL ) Was stirred at room temperature for 2 hours. The mixture was concentrated in vacuo and EA (150 mL) was added. The pH of the aqueous layer was adjusted to 4 by the addition of 1N HCl solution and the aqueous layer was extracted with EA (2 × 150 mL). The combined organic layers were dried over Na ₂ SO ₄ , filtered and concentrated in vacuo. The resulting title compound was used in the next reaction without purification. MS (m / z): 238.8 [M + H ^<+ >]

C. [5- (4-Pyridin-4-yl-piperazin-1-carbonyl) -pyridin-2-yl] -carbamic acid tert-butyl ester 6-tert-butoxycarbonylamino-nicotinic acid (90 mg in DMF (3 mL) 1-pyridin-4-yl-piperazine (123 mg, 0.75 mmol) was added to a stirred solution of WSC (86 mg, 0.45 mmol) and HOBt (61 mg, 0.45 mmol). It was. After stirring at room temperature for 3 hours, the solvent was removed in vacuo and the residue was purified by reverse phase HPLC using a gradient of ACN in water containing 0.1% TFA to give the title compound as a TFA salt. MS (m / z): 384.0 [M + H ⁺ ]

D. (6-Amino-pyridin-3-yl)-(4-pyridin-4-yl-piperazin-1-yl) -methanone [5- (4-Pyridin-4-yl-piperazine-1 in DCM (5 mL) A solution of -carbonyl) -pyridin-2-yl] -carbamic acid tert-butyl ester (24 mg, 0.06 mmol) and TFA (5 mL) was stirred at room temperature for 4 hours. The reaction mixture was concentrated in vacuo to give the title compound as a TFA salt. MS (m / z): 284.2 [M + H ⁺ ]

E. 4- (2,5-cis / trans-dimethyl-pyrrolidine-1-carbonyl) -2-fluoro-benzaldehyde [5- (4-pyridin-4-yl-piperazin-1-carbonyl) -pyridin-2-yl] -According to the synthesis of carbamic acid tert-butyl ester, 3-fluoro-4-formyl-benzoic acid (46 mg, 0.28 mmol) with 2,5-cis / trans-dimethyl-pyrrolidine (34 [mu] L, 0.28 mmol). Reacted to give the title compound as a TFA salt. MS (m / z): 250.1 [M + H ⁺ ]

F. {6- [4- (2,5-cis / trans-dimethyl-pyrrolidin-1-carbonyl) -2-fluoro-benzylamino] -pyridin-3-yl}-(4-pyridin-4-yl-piperazine- 1-yl) -methanone (6-Amino-pyridin-3-yl)-(4-pyridin-4-yl-piperazin-1-yl) -methanone (17.5 mg, 0.062 mmol) in dichloroethane (2 mL) ), 4- (2,5-cis / trans-dimethyl-pyrrolidine-1-carbonyl) -2-fluoro-benzaldehyde (11.0 mg, 0.044 mmol) and titanium-tetra-isopropylate (62.5 mg, 0 .220 mmol) was stirred at room temperature overnight. Sodium triacetoxyborohydride (18.6 mg, 0.088 mmol) was added and the mixture was stirred at room temperature for an additional 3 days. The solvent was removed in vacuo and the residue was partitioned between EA (20 mL) and saturated NaHCO ₃ solution (20 mL). The aqueous layer was extracted with EA (2 × 20 mL) and the organic layer was dried over Na ₂ SO ₄ , filtered and concentrated in vacuo. The residue was purified by reverse phase HPLC using a gradient of ACN in water containing 0.1% TFA to give the title compound as a TFA salt. MS (m / z): 517.3 [M + H ^<+ >]

{6- [4- (2,5-cis / trans-dimethyl-2,5-dihydro-pyrrol-1-carbonyl) -2-fluoro-benzylamino] -pyridin-3-yl}-(4-pyridine- Synthesis of 4-yl-piperazin-1-yl) -methanone (compound 111)

A. 3-fluoro-4-{[5- (4-pyridin-4-yl-piperazin-1-carbonyl) -pyridin-2-ylamino] -methyl} -benzoic acid methyl ester {6- [4- (2,5 According to the synthesis of -cis / trans-dimethyl-pyrrolidin-1-carbonyl) -2-fluoro-benzylamino] -pyridin-3-yl}-(4-pyridin-4-yl-piperazin-1-yl) -methanone (6-Amino-pyridin-3-yl) (4-pyridin-4-yl-piperazin-1-yl) -methanone (92 mg, 0.32 mmol) was converted to 3-fluoro-4-formyl-benzoic acid methyl ester ( 65 mg, 0.35 mmol) to give the title compound, which was used in the subsequent step without further purification. MS (m / z): 450.3 [M + H ⁺ ]

B. {6- [4- (2,5-cis / trans-dimethyl-2,5-dihydro-pyrrol-1-carbonyl) -2-fluoro-benzylamino] -pyridin-3-yl}-(4-pyridine- 4-yl-piperazin-1-yl) -methanone According to the synthesis of 3-fluoro-4-formyl-benzoic acid, 3-fluoro-4-{[5- (4-pyridin-4-yl-piperazine-1-carbonyl ) -Pyridin-2-ylamino] -methyl} -benzoic acid methyl ester (110 mg, 0.23 mmol) was reacted with LiOH to give 3-fluoro-4-{[5- (4-pyridin-4-yl-piperazine -1-carbonyl) -pyridin-2-ylamino] -methyl} -benzoic acid was provided. [5- (4
-Pyridin-4-yl-piperazin-1-carbonyl) -pyridin-2-yl] -carbamic acid According to the synthesis of tert-butyl ester, benzoic acid (18 mg, 0.04 mmol) was converted to 2,5-cis / trans- Reaction with dimethyl-2,5-dihydro-1H-pyrrolidine (25 μL, 0.20 mmol) gave the title compound as the TFA salt. MS (m / z): 515.2 [M + H ^<+ >]

Synthesis of (R) -pyrimidine-5-carboxylic acid (1- {1- [4- (ethyl-isopropyl-carbamoyl) -phenyl] -ethylcarbamoyl} -1-methyl-ethyl) -amide (Compound 115)

A. (R) -4- [1- (9H-Fluoren-9-ylmethoxycarbonylamino) -ethyl] -benzoic acid (R) -4- (1-amino-ethyl) -benzoic acid in DCM (50 mL) ( To a solution of 2.00 g, 9.92 mmol) was added trimethylsilyl chloride (3.14 mL, 24.8 mmol). The mixture was heated at reflux for 2 hours, then cooled to room temperature and triethylamine (5.53 mL, 39.7 mmol) was added dropwise, followed by 9-fluorenylmethyl chloroformate (2.82 g, 10.9 mmol). It was dripped. The reaction mixture was stirred at room temperature overnight and then concentrated in vacuo. The residue was stirred in saturated sodium carbonate solution and acidified with 2N hydrochloric acid. The resulting aqueous solution was extracted with diethyl ether (2 × 150 mL) and the organic layer was dried over Na ₂ SO ₄ , filtered and concentrated in vacuo to give the title compound. MS (m / z): 387.7 [M + H ^<+ >]

B. Resin-bonded (R) -4- (1-amino-ethyl) -benzoic acid 4- [1- (9H-fluoren-9-ylmethoxycarbonylamino) -ethyl] -benzoic acid (296 mg, 0.8 mmol) And DIPEA (332 μL, 1.9 mmol) were dissolved in DCM (8 mL). The solution was added to 2-chlorotrityl chloride resin (loading: 1.4 mmol / g, 1.09 g, 1.5 mmol) swollen in DCM (8 mL) and the mixture was shaken at room temperature for 30 min. The mixture was quenched with methanol (800 μL). The resin was filtered and washed with DCM / methanol / DIPEA (16: 3: 3, 1 × 10 mL) and DMF (3 × 10 mL). Piperidine (25% solution in DMF, 10 mL) was added and the mixture was shaken for 20 minutes. The resin was filtered, washed with DMF (6 × 10 mL) and dried in vacuo to give the title compound at a predicted loading of 45% (0.65 mmol / g).

C. Resin-bonded (R) -4- [1- (2-amino-2-methyl-propionylamino) -ethyl] -benzoic acid 2- (9H-fluoren-9-ylmethoxycarbonylamino) in DMF (1 mL) A solution of 2-methyl-propionic acid (63 mg, 0.19 mmol), HATU (74 mg, 0.19 mmol) and DIPEA (68 μL, 0.39 mmol) was stirred at room temperature for 5 minutes, then DCM (1 mL) Resin-bonded (R) -4- (1-amino-) swollen in
Ethyl) -benzoic acid (100 mg, 0.07 mmol). The mixture was shaken at room temperature for 1.5 hours and then quenched with methanol. The resin was filtered and washed with DMF (4 × 2 mL) to give resin-bound 4- {1- [2- (9H-fluoren-9-ylmethoxycarbonylamino) -2-methyl-propionylamino] -ethyl} -benzoic acid. Acid was given. Test cleavage with TFA showed the correct product. MS (m / z): 472.9 [M + H ^<+ >]. Piperidine (25% solution in DMF, 4 mL) was added and the mixture was shaken for 20 minutes. The resin was filtered, washed with DMF (6 × 10 mL) and dried in vacuo to give the title compound.

D. Resin-bonded (R) -4- (1- {2-methyl-2-[(pyrimidine-5-carbonyl) -amino] -propionylamino} -ethyl) -benzoic acid Resin-bonded 4- {1- [2 According to the synthesis of-(9H-fluoren-9-ylmethoxycarbonylamino) -2-methyl-propionylamino] -ethyl} -benzoic acid, resin-bonded (R) -4- [1- (2-amino-2- Methyl-propionylamino) -ethyl] -benzoic acid (100 mg, 0.065 mmol) was reacted with 5-pyrimidinecarboxylic acid. Test cleavage with TFA showed the correct product. MS (m / z): 356.9 [M + H ^<+ >]

E. (R) -4- (1- {2-methyl-2-[(pyrimidine-5-carbonyl) -amino] -propionylamino} -ethyl) -benzoic acid 1,1,1,3,3,3-hexa Resin-bonded (R) -4- (1- {2-methyl-2-[(pyrimidine-5-carbonyl) -amino] -propionyl in fluoro-2-propanol / DCM (4: 1, 2.5 mL) A suspension of amino} -ethyl) -benzoic acid (100 mg, 0.065 mmol) was shaken at room temperature for 3 hours. The resin was filtered and washed with DCM (2 × 3 mL). The filtrate was concentrated in vacuo to give the title compound. MS (m / z): 356.9 [M + H ^<+ >]

F. (R) -pyrimidine-5-carboxylic acid (1- {1- [4- (ethyl-isopropyl-carbamoyl) -phenyl] -ethylcarbamoyl} -1-methyl-ethyl) -amide in DMF (0.4 mL) (R) -4- (1- {2-methyl-2-[(pyrimidine-5-carbonyl) -amino] -propionylamino} -ethyl) -benzoic acid (12.0 mg, 0.034 mmol) and HATU ( To a stirred solution of 12.8 mg (0.034 mmol) was added N-ethylisopropylamine (41 μL, 0.337 mmol). After stirring overnight, the reaction mixture was concentrated in vacuo and the residue was purified by reverse phase HPLC using a gradient of ACN in water containing 0.1% TFA to give the title compound as a TFA salt. MS (m / z): 426.1 [M + H ^<+ >]

(R) -N- (cyclopropyl- {1- [4- (2,5-cis / trans-dimethyl-pyrrolidine-1-carbonyl) -phenyl] -ethylcarbamoyl} -methyl) -3-fluoro-5- Synthesis of trifluoromethyl-benzamide (Compound 116)

A. Resin-bonded (R) -4- [1- (2-amino-2-cyclopropyl-acetylamino) -ethyl] -benzoic acid 4- {1- [2- (9H-fluoren-9-ylmethoxycarbonylamino] According to the synthesis of) -2-methyl-propionylamino] -ethyl} -benzoic acid, resin-bound (R) -4- (1-amino-ethyl) -benzoic acid (58.7 mg, 0.038 mmol) is ( Reaction with R) -cyclopropyl-[(9H-fluoren-9-ylmethoxycarbonylamino)]-acetic acid (38.6 mg, 0.114 mmol). Test cleavage with TFA showed the correct product 4- {1- [2-cyclopropyl-2- (9H-fluoren-9-ylmethoxycarbonylamino) -acetylamino] -ethyl} -benzoic acid. MS (m / z): 485.0 [M + H ^<+ >]. Deprotection with piperidine following the synthesis of resin-bound (R) -4- [1- (2-amino-2-methyl-propionylamino) -ethyl] -benzoic acid gave the title compound.

B. (R) -N- (cyclopropyl- {1- [4- (2,5-cis / trans-dimethyl-pyrrolidine-1-carbonyl) -phenyl] -ethylcarbamoyl} -methyl) -3-fluoro-5- Trifluoromethyl-benzamide 4- {1- [2- (9H-fluoren-9-ylmethoxycarbonylamino) -2-methyl-propionylamino] -ethyl} -benzoic acid according to the synthesis of resin-bonded (R)- 4- [1- (2-Amino-2-cyclopropyl-acetylamino) -ethyl] -benzoic acid (58.7 mg, 0.038 mmol) was converted to 3-fluoro-5-trifluoromethylbenzoic acid (15.8 mg). , 0.076 mmol). According to the synthesis of (R) -4- (1- {2-methyl-2-[(pyrimidine-5-carbonyl) -amino] -propionylamino} -ethyl) -benzoic acid (12 mg, 0.034 mmol) 1, Cleavage with 1,1,3,3,3-hexafluoro-2-propanol / DCM is 4- {1- [2-cyclopropyl-2- (3-fluoro-5-trifluoromethyl-benzoylamino) -Acetylamino] -ethyl} -benzoic acid was given. (R) -Pyrimidine-5-carboxylic acid Acid according to the synthesis of (1- {1- [4- (ethyl-isopropyl-carbamoyl) -phenyl] -ethylcarbamoyl} -1-methyl-ethyl) -amide (12.5 mg , 0.028 mmol) of 2,5-cis / trans-dimethylpyrrolidine (6.8 μL, 0.055 mmol) gave the title compound as the TFA salt. MS (m / z): 534.1 [M + H ^<+ >]

(R, S) -N- (1- {1- [2-Chloro-4-((2R, 5R) -2,5-dimethyl-pyrrolidine-1-carbonyl) -phenyl] -ethylcarbamoyl} -cyclopropyl ) Synthesis of -3-fluoro-5-trifluoromethyl-benzamide (Compound 100)

A. 3-Chloro-4-methyl-benzoic acid methyl ester To a stirred solution of 3-chloro-4-methyl-benzoic acid (2.0 g, 11.7 mmol) in methanol (40 mL) was added chloro-trimethyl-silane (6 0.0 mL, 46.9 mmol) was added. After stirring at room temperature overnight, the solvent was removed in vacuo to give the title compound, which was used in the subsequent step without further purification. GC / MS (m / z): 184

B. 3-Chloro-4-formyl-benzoic acid methyl ester 3-chloro-4-methyl-benzoic acid methyl ester (1.04 g, 5.61 mmol), 1-bromo-pyrrolidine-2 in tetrachloromethane (68 mL) , 5-dione (3.26 g, 15.7 mmol) and a suspension of benzoyl peroxide (100.2 mg, 0.88 mmol) after stirring at 80 ° C. overnight, the mixture was concentrated in vacuo, The residue was partitioned between EA (300 mL) and water (200 mL). The organic layer was washed with saturated NaHCO ₃ solution (1 × 200 mL), water (1 × 200 mL) and saturated NaCl solution (1 × 200 mL). The organic layer was dried over Na ₂ SO ₄ , filtered and concentrated in vacuo. A mixture of the residue, 4-bromomethyl-3-chloro-benzoic acid methyl ester and 4-dibromomethyl-3-chloro-benzoic acid methyl ester, was used in the subsequent step without further purification. The residue (1.48 g, 5.61 mmol) was dissolved in acetone (45 mL) and water (9 mL) and AgNO ₃ (3.22 g, 16.3 mmol) was added. The flask was covered with aluminum foil to avoid decomposition of AgNO ₃ . The mixture was stirred at room temperature overnight. After filtration of the mixture and evaporation of the solvent, the residue was partitioned between EA (700 mL) and saturated NaHCO ₃ solution (350 mL). The organic layer was washed with water (1 × 350 mL) and saturated NaCl solution (1 × 350 mL), dried over Na ₂ SO ₄ , filtered and concentrated in vacuo. The residue was purified by flash chromatography on silica gel (eluting with n-hexane / EA 9: 1) to give the title compound. GC / MS (m / z): 198

C. Synthesis of (R, S) -4- (1-azido-ethyl) -3-chloro-benzoic acid methyl ester (R, S) -3-fluoro-4- (1-hydroxy-ethyl) -benzoic acid methyl ester 3-chloro-4-formyl-benzoic acid methyl ester (398 mg, 2.01 mmol) was reacted with methylmagnesium bromide according to Reaction of the ester with methanesulfonyl chloride followed by reaction with sodium azide following the synthesis of (R, S) -4- (1-azido-ethyl) -3-fluoro-benzoic acid methyl ester gave the title compound. GC / MS (m / z): 239

D. Synthesis of (R, S) -4- (1-amino-ethyl) -3-chloro-benzoic acid methyl ester (R, S) -4- (1-amino-ethyl) -3-fluoro-benzoic acid methyl ester (R, S) -4- (1-azido-ethyl) -3-chloro-benzoic acid methyl ester (181 mg, 0.76 mmol) was added according to THales H-Cube ^(R)
Subjected to continuous flow hydrogenation using a flow hydrogenerator to give the title compound. MS (m / z): 213.8 [M + H ^<+ >]

E. (R, S) -3-chloro-4- (1-{[1- (3-fluoro-5-trifluoromethyl-benzoylamino) -cyclopropanecarbonyl] -amino} -ethyl) -benzoic acid methyl ester ( R, S) -4- (1-amino-ethyl) -3-chloro-benzoic acid methyl ester (17.7 mg, 0.83 mmol) is reacted with 1-tert-butoxycarbonylamino-cyclopropanecarboxylic acid, It was then treated with TFA according to the synthesis of (R) -4- {1-[(1-amino-cyclopropanecarbonyl) -amino] -ethyl} -benzoic acid methyl ester. The following (R) -4- [1-({1-[(pyrimidine-5-carbonyl) -amino] -cyclopropanecarbonyl} -amino) -ethyl] -benzoic acid methyl ester follows the synthesis of 3-fluoro-5- Reaction with trifluoromethyl-benzoic acid gave the title compound. MS (m / z): 472.9 [M + H ^<+ >].

F. (R, S) -N- (1- {1- [2-Chloro-4-((2R, 5R) -2,5-dimethyl-pyrrolidine-1-carbonyl) -phenyl] -ethylcarbamoyl} -cyclopropyl ) -3-Fluoro-5-trifluoromethyl-benzamide (R, S) -3-chloro-4- (1-{[1- (3-fluoro-5-trifluoromethyl-benzoylamino) -cyclopropanecarbonyl ] -Amino} -ethyl) -benzoic acid methyl ester (40.3 mg, 0.083 mmol) was treated with LiOH (1.4 ml) to give (R, S) -3-chloro-4- (1-{[ 1- (3-Fluoro-5-trifluoromethyl-benzoylamino) -cyclopropanecarbonyl] -amino} -ethyl) -benzoic acid was given. Followed by (R, S) -pyrimidine-5-carboxylic acid (1- {1- [4- (2,5-cis / trans-dimethyl-pyrrolidine-1-carbonyl) -2-fluoro-phenyl] -ethylcarbamoyl} Reaction of benzoic acid with (2R, 5R) -2,5-dimethyl-pyrrolidine following the synthesis of -cyclopropyl) -amide gave the title compound as the TFA salt. MS (m / z): 554.0 [M + H ⁺ ]

(R, S) -N- (1- {1- [2-Chloro-4-((2S, 5S) -2,5-dimethyl-pyrrolidine-1-carbonyl) -phenyl] -ethylcarbamoyl} -cyclopropyl ) Synthesis of 3-Fluoro-5-trifluoromethyl-benzamide (Compound 99)

  Reaction AD was performed based on the literature (Schlessinger et al, work, Tetrahedron Lett. 28, 1987, 2083-2086).

A. (2-Hydroxy-1 (S) -methyl-ethyl) -carbamic acid benzyl ester Lithium aluminum hydride (10.2 g, 0.27 mol) was suspended in anhydrous THF (450 mL). To this mixture, L-alanine (12.0 g, 0.13 mol) was added in portions at 0 ° C. The mixture was refluxed overnight and then cooled to 0 ° C. 2N NaOH solution (70 mL) was added. After stirring at room temperature for 3 hours, the mixture was filtered and the solid was washed with THF. The solid was suspended in THF (250 mL) and the mixture was refluxed for 1 hour. The solution was filtered and the solid was washed with THF. This procedure was repeated twice. To the combined THF solution was added 2N NaOH solution (170 mL) and benzyl chloroformate (25.3 g, 0.15 mol).
Was added. After stirring for 1 hour, the two-phase system was separated and the aqueous layer was extracted with EA (1 × 50 mL). The organic layer was dried over Na ₂ SO ₄ , filtered and concentrated in vacuo. Crystallization of the crude compound from THF / cyclohexane gave the title compound.

B. (S) -Toluene-4-sulfonic acid 2-benzyloxycarbonylamino-propyl ester (2-hydroxy-1 (S) -methyl-ethyl) -carbamic acid benzyl ester (12.0 g, 0) in pyridine (190 mL) 0.06 mol) was added p-toluenesulfonyl chloride (4.40 g, 0.11 mol) at 0 ° C. After stirring at room temperature overnight, diethyl ether (50 mL) was added. The mixture was filtered and the solid was washed with diethyl ether. The organic extract is washed with 0.5N H ₂ SO ₄ solution (3 × 20 mL), 5% NaHCO ₃ solution (1 × 20 mL) and saturated NaCl solution (1 × 20 mL), dried over Na ₂ SO ₄ , filtered and in vacuo. Concentrated. Crystallization of the crude compound from THF / n-hexane gave the title compound.

C. (S) -Toluene- (2-iodo-1-methyl-ethyl) -carbamic acid benzyl ester (S) -Toluene-4-sulfonic acid 2-benzyloxycarbonylamino-propyl ester in acetone (113 mL) (12. To a solution of 3 g, 0.03 mol), solid sodium iodide (50.7 g, 0.34 mol) was added at 0 ° C. After stirring at 0 ° C. for 30 minutes, the reaction mixture was stirred at room temperature for 48 hours. The solvent was removed in vacuo at room temperature. The residue was suspended in EA. The mixture was filtered and the solid was washed with EA. The organic extract is washed with H ₂ O ( ₂ × 20 mL), 5% Na ₂ S ₂ O ₃ solution (1 × 20 mL) and saturated NaCl solution (1 × 20 mL), dried over Na ₂ SO ₄ , filtered and concentrated in vacuo. did. Crystallization of the crude compound from THF / n-hexane gave the title compound.

D. (S)-(1-Methyl-pent-4-enyl) -carbamic acid benzyl ester To a stirred suspension of copper iodide (3.60 g, 0.019 mol) in anhydrous THF (64 mL) was added -78 ° C. Allylmagnesium chloride (19.0 mL, 0.038 mol, 2M solution in THF) was added dropwise. When the addition was complete, the mixture was warmed to −36 ° C. for 5 minutes, then cooled to −78 ° C. and benzyl (S) -toluene- (2-iodo-1-methyl-ethyl) -carbamate in THF (19 mL). Ester (4.00 g, 0.013 mol) was added dropwise. After stirring at −36 ° C. for 6 hours, saturated NH ₄ Cl solution (10 mL) was added, allowing the mixture to warm to room temperature. The solvent was removed in vacuo. The residue was suspended in diethyl ether. The mixture was filtered and the solid was washed with diethyl ether. The organic extract was washed with saturated NaCl solution (1 × 20 mL), NH ₄ OH solution (3 × 20 mL) and saturated NaCl solution (1 × 20 mL), dried over Na ₂ SO ₄ , filtered and concentrated in vacuo. The residue was purified by flash chromatography on silica gel (eluting with n-hexane / diethyl ether 1: 1) to give the title compound. MS (m / z): 233.8 [M + H ^<+ >]

  Reaction E was performed based on literature (Harding et al., J. Org. Chem. 46, 1981, 3920-3922).

E. (2S, 5S) -2,5-Dimethyl-pyrrolidine-1-carboxylic acid benzyl ester (S)-(1-methyl-pent-4-enyl) -carbamic acid benzyl ester (1.00 g) in THF (43 mL) , 4.29 mmol) was added mercury acetate (2.05 g, 6.43 mmol). The mixture was purged with nitrogen, covered with aluminum foil and stirred at room temperature overnight. Sodium borohydride (1) in 2N NaOH (0.32 mL)
60 mg, 4.24 mmol) was added dropwise and the mixture was stirred at room temperature overnight. Saturated Na ₂ CO ₃ solution (3 mL) was added. After stirring at room temperature for 4 hours, the solvent was removed in vacuo. The residue was diluted with diethyl ether and extracted with saturated Na ₂ CO ₃ solution ( ₃ × 15 mL). The aqueous layer was extracted with diethyl ether (3 × 20 mL). The organic layer was dried over Na ₂ SO ₄ , filtered and concentrated in vacuo. The residue was purified by flash chromatography on silica gel (eluting with DCM / methanol 30: 1) to give the title compound. MS (m / z): 233.8 [M + H ^<+ >]

F. (2S, 5S) -2,5-Dimethyl-pyrrolidine (2S, 5S) -2,5-Dimethyl-pyrrolidine-1-carboxylic acid benzyl ester (100 mg, 0.43 mmol) was dissolved in methanol (9 mL). . With Thales H-Cube ^(R) flow Hydrogenics Noether containing cartridge 10% palladium on carbon as catalyst, the solution was subjected to continuous flow hydrogenation of. Hydrogenation was carried out at room temperature under a pressure of 10 bar. Twenty drops of concentrated HCl was added. The solvent was evaporated to give the title compound as the HCl salt. MS (m / z): 100.0 [M + H ⁺ ]

G. (R, S) -N- (1- {1- [2-chloro-4-((2S, 5S) -2,5-dimethyl-pyrrolidine-1-carbonyl) -phenyl] -ethylcarbamoyl} -cyclopropyl ) -3-Fluoro-5-trifluoromethyl-benzamide (R, S) -pyrimidine-5-carboxylic acid (1- {1- [4- (2,5-cis / trans-dimethyl-pyrrolidine-1-carbonyl) ) -2-Fluoro-phenyl] -ethylcarbamoyl} -cyclopropyl) -amide according to the synthesis of (R, S) -3-chloro-4- (1-{[1- (3-fluoro-5-trifluoro Methyl-benzoylamino) -cyclopropanecarbonyl] -amino} -ethyl) -benzoic acid is reacted with (2S, 5S) -2,5-dimethyl-pyrrolidine to give the title compound as a TFA salt I was painting. MS (m / z): 554.0 [M + H ⁺ ]

N-((1R) -1- (4- (2,5-cis / trans-dimethylpyrrolidine-1-carbonyl) phenyl) ethyl) -2- (3-fluoro-5- (trifluoromethyl) benzoyl) hydrazine Synthesis of Carboxamide (Compound 150)

  Based on the literature, the first stage of reaction A was carried out (Gibson et al., J. Org. Chem. 64, 1999, 7388-7394).

A. Resin-bound (R) -4- (1- (hydrazinecarboxamido) ethyl) benzoic acid hydrazinecarboxylic acid 9H-fluoren-9-ylmethyl ester (150 mg, in DCM (5 mL) and saturated NaHCO ₃ solution (5 mL) 0.59 mmol) was suspended. The biphasic mixture was stirred at 0 ° C. for 5 minutes. The two layers were separated and phosgene (20% solution in toluene, 1.3 mL, 2.95 mmol) was added to the organic phase via syringe. The mixture was stirred at 0 ° C. for 10 minutes, the layers were separated and the aqueous layer was extracted with DCM (3 × 10 mL). The combined organic layers were dried over _Na 2 SO _4, filtered, and concentrated in vacuo, 5-(9H-fluoren-9-ylmethoxy) -3H- [1,3,4] oxadiazol-2 -Given ON. The crude product (350 mg, 0.483 mmol) was dissolved in DCM (2 mL) and swollen in DCM (5 mL) and collidine (136 μL, 1.035 mmol) resin-bound (R) -4- ( To a suspension of 1-amino-ethyl) -benzoic acid (106 mg, 0.069 mmol). The mixture was shaken for 90 minutes at room temperature. The resin was filtered and washed with DMF (4 × 2 mL), methanol (4 × 2 mL) and DCM (4 × 2 mL). Test cleavage with TFA showed the correct product (R) -4- (1- (2-(((9H-fluoren-9-yl) methoxy) carbonyl) hydrazinecarboxamido) ethyl) benzoic acid. MS (m / z): 445.8 [M + H ^<+ >]. Piperidine (20% solution in DMF, 4 mL) was added and the mixture was shaken for 15 minutes. The resin was filtered, washed with DMF (4 × 2 mL), methanol (4 × 2 mL) and DCM (4 × 2 mL) and dried in vacuo. Test cleavage with TFA showed the correct product. MS (m / z): 223.8 [M + H ^<+ >]

B. (R) -4- (1- (2- (3-Fluoro-5- (trifluoromethyl) benzoyl) hydrazinecarboxamido) ethyl) benzoic acid 3-fluoro-5-trifluoromethyl-in DCM (1 mL) A solution of benzoic acid (17.2 mg, 0.083 mol), HATU (31.5 mg, 0.083 mmol) and collidine (45.4 μL, 0.345 mmol) was added to resin-bound (R) -4- ( 1- (hydrazinecarboxamido) ethyl) benzoic acid (106 mg, 0.069 mmol) was added. The mixture was shaken for 2 hours at room temperature. The resin was filtered and washed with DMF (4 × 2 mL), methanol (4 × 2 mL) and DCM (4 × 2 mL) and suspended in DCM (1 mL) and TFA (1 mL). The suspension was shaken for 30 minutes at room temperature. The resin was filtered and washed with DCM (2 × 2 mL). The filtrate was concentrated in vacuo. The residue was purified by reverse phase HPLC using a gradient of ACN in water containing 0.1% TFA to give the title compound. MS (m / z): 413.8 [M + H ^<+ >]

C. N-((1R) -1- (4- (2,5-cis / trans-dimethylpyrrolidine-1-carbonyl) phenyl) ethyl) -2- (3-fluoro-5- (trifluoromethyl) benzoyl) hydrazine Carboxamide DCM / DMF 9: 1 (R) -4- (1- (2- (3-Fluoro-5- (trifluoromethyl) benzoyl) hydrazinecarboxamido) ethyl) benzoic acid (16. 2 mg, 0.039 mmol), WSC (9.0 mg, 0.047 mmol) and collidine (25.8 μL, 0.196 mmol) in a solution of 2,5-cis / trans-dimethyl-pyrrolidine (4.7 mg). 0.047 mmol) was added. After stirring at room temperature overnight, the mixture was concentrated in vacuo. The residue was purified by reverse phase HPLC using a gradient of ACN in water containing 0.1% TFA to give the title compound. MS (m / z): 495.1 [M + H ^<+ >]

N-((1R) -1- (4- (2,5-cis / trans-dimethylpyrrolidine-1-carbonyl) phenyl) ethyl) -2- (3-fluoro-5- (trifluoromethyl) benzoyl)- Synthesis of 1-methylhydrazinecarboxamide (Compound 151)

  Based on the literature, the first stage of reaction A and reaction B was carried out (Gibson et al., J. Org. Chem. 64, 1999, 7388-7394).

A. N-methyl-hydrazinecarboxylic acid 9H-fluoren-9-ylmethyl ester To a stirred solution of di-tert-butyl dicarbonate (924 mg, 4.23 mmol) in DCM (5 mL) was added methylhydrazine (173 μL, 3.26). Mmol) was added at -78 ° C. The solution was allowed to warm to room temperature and stirred at room temperature for 30 minutes. 9H-fluorenylmethyl chloroformate (842 mg, 3.26 mmol) and ethyl-diisopropyl-amine (664 μL, 3.91 mmol) were added dropwise and the mixture was stirred at room temperature for 8 hours. TFA (5 mL) was added and the mixture was stirred for an additional 2 hours. The mixture was concentrated in vacuo and the residue was purified by flash chromatography on silica gel (eluting with n-hexane / EA 7: 3) to give the title compound. MS (m / z): 268.8 [M + H ⁺ ]

B. (R) -4- (1- (2- (3-Fluoro-5- (trifluoromethyl) benzoyl) -1-methylhydrazinecarboxamido) ethyl) benzoic acid N-methyl-hydrazinecarboxylic acid in dioxane (15 mL) To a stirred solution of acid 9H-fluoren-9-ylmethyl ester (1.81 g, 3.26 mmol) phosgene (20% solution in toluene, 3.43 ml, 6.51 mmol) was added dropwise at 10 ° C. did. The mixture was stirred at 10 ° C. for 5 minutes and then at room temperature for 1.5 hours. The mixture was concentrated in vacuo to give 2- (chlorocarbonyl) -2-methylhydrazinecarboxylic acid (9H-fluoren-9-yl) methyl. The crude product (68.5 mg, 0.207 mmol) was dissolved in DCM (1 mL) and swollen in DCM (5 mL) and collidine (136 μL, 1.035 mmol). Resin-bond (R) -4 To a suspension of-(1-amino-ethyl) -benzoic acid (106 mg, 0.069 mmol). The mixture was shaken for 90 minutes at room temperature. The resin was filtered and washed with DMF (4 × 2 mL), methanol (4 × 2 mL) and DCM (4 × 2 mL). Test cleavage with TFA yields the correct product (R) -4- (1- (2-(((9H-fluoren-9-yl) methoxy) carbonyl) -1-methylhydrazinecarboxamido) ethyl) benzoic acid. Indicated. MS (m / z): 459.8 [M + H ^<+ >]. Piperidine (20% solution in DMF, 4 mL) was added and the mixture was shaken for 30 minutes to give resin-bound (R) -4- (1- (1-methylhydrazinecarboxamido) ethyl) benzoic acid. . The resin was filtered, washed with DMF (4 × 2 mL), methanol (4 × 2 mL) and DCM (4 × 2 mL) and 3-fluoro-5-trifluoromethyl-benzoic acid (17 in DCM / DMF 2: 1 (1.5 ml). .2 mg, 0.083 mmol), HATU (31.5 mg, 0.083 mmol) and collidine (45.4 μL, 0.345 mmol) were added. The mixture was shaken for 4 hours at room temperature. The resin was filtered and washed with DMF (4 × 2 mL), methanol (4 × 2 mL) and DCM (4 × 2 mL) and suspended in DCM (1 mL) and TFA (1 mL). The suspension was shaken for 20 minutes at room temperature. The resin was filtered and washed with DCM (2 × 2 mL). The filtrate was concentrated in vacuo. The residue was purified by reverse phase HPLC using a gradient of ACN in water containing 0.1% TFA to give the title compound. MS (m / z): 427.7 [M + H ⁺ ]

C. N-((1R) -1- (4- (2,5-cis / trans-dimethylpyrrolidine-1-carbonyl) phenyl) ethyl) -2- (3-fluoro-5- (trifluoromethyl) benzoyl)- 1-methylhydrazinecarboxamide (R) -4- (1- (2- (3-fluoro-5- (trifluoromethyl) benzoyl) -1-methylhydrazinecarboxamide) in DCM / DMF 9: 1 (1 mL) ) Ethyl) benzoic acid (10 mg, 0.023 mmol), WSC (5.4 mg, 0.028 mmol) and collidine (15.4 μL, 0.117 mmol) in a solution of 2,5-cis / trans-dimethyl. -Pyrrolidine (2.8 mg, 0.028 mmol) was added. After stirring at room temperature overnight, the mixture was concentrated in vacuo. The residue was purified by reverse phase HPLC using a gradient of ACN in water containing 0.1% TFA to give the title compound. MS (m / z): 509.0 [M + H ⁺ ]

N-((R) -1- (5-((2R, 5S) -2,5-dimethylpyrrolidin-1-carbonyl) -3-fluoropyridin-2-yl) ethyl) -1-ethyl-2- ( Synthesis of 3-fluoro-5- (trifluoromethyl) benzoyl) hydrazinecarboxamide (Compound 268)

A. 5-Bromo-3-fluoro-pyridine-2-carbonitrile To a solution of TBAF in THF (657 ml, 0.66 mol, 1M) was added H ₂ SO ₄ (1 ml, 5 μmol). DMF (200 mL) was added at −40 ° C. until the suspension became a clear solution. 5-Bromo-3-nitropyridine-2-carbonitrile (50.0 g, 0.22 mol) in DMF (800 mL) was added slowly. The solution was stirred at −40 ° C. for 30 minutes until no starting material could be detected by TLC (n-hexane / EA 5: 1). The reaction was quenched with 2M HCl (500 mL) at −40 ° C. to give pH 3. The mixture was extracted twice with EA. The combined organic layers were washed with water, dried over sodium sulfate and concentrated in vacuo. The crude product was purified by chromatography on silica gel.

B. 5-Bromo-3-fluoro-pyridine-2-carboxylic acid Concentrated HCl (300 mL) was added to 5-bromo-3-fluoro-pyridine-2-carbonitrile (21.0 g, 0.11 mol) at 60 ° C. Stir overnight. After evaporation, ether was added and the suspension was stirred for an additional time. The solid was collected and dried. The crude product was used without further purification. MS (m / z): 219.8 [M + H ^<+ >]

C. 5-Bromo-3-fluoro-pyridine-2-carboxylic acid Methoxy-methyl-amide of 5-bromo-3-fluoro-pyridine-2-carboxylic acid (55.5 g, 0.22 mol) in DMF (1000 mL) To the solution, TEA (17.4 ml, 0.13 mol), EDCI (83.2 g, 0.43 mol), HOBt (2.90 g, 0.02 mol) and N, O-
Dimethylhydroxylamine xHCl (25.0 g, 0.26 mol) was added. After stirring overnight, the solvent was reduced in vacuo. EA and brine were added and the aqueous layer was extracted with EA. The combined organic layers were washed with brine, dried over sodium sulfate and the solvent removed under reduced pressure. The crude product was purified by chromatography on silica gel. MS (m / z): 262.8 [M + H ^<+ >]

D. 5-Bromo-3-fluoro-pyridine-2-carbaldehyde of 5-bromo-3-fluoro-pyridine-2-carboxylic acid methoxy-methyl-amide (29.0 g, 0.11 mol) in THF (350 mL) To the solution was slowly added 1M LiAlH _{4 in} THF (50 ml, 0.05 mol) at −78 ° C. After stirring at −78 ° C. for 20 minutes, the mixture was quenched with water and brine and allowed to reach room temperature. EA was added and the suspension was filtered over celite. The aqueous layer was extracted with EA and the combined organic layers were washed with brine and dried over sodium sulfate. After evaporation, the crude product was purified by chromatography on silica gel (n-hexane / EA). MS (m / z): 203.9 [M + H ^<+ >].

E. 2-Methyl-propane-2-sulfinic acid 5-Bromo-3-fluoro-pyridin-2-ylmethyleneamide 5-Bromo-3-fluoro-pyridine-2-carbaldehyde (19.5 g, in DCM (600 ml)) 0.10 mol) solution was added (R)-(+)-2-methyl-2-propanesulfinamide (11.6 g, 0.10 mol), magnesium sulfate (115 g, 0.10 mol), PPTS ( 2.40 g, 0.01 mol) and cesium carbonate (33.7 g, 0.10 mol) were added. After stirring overnight, the reaction was filtered over celite and the solvent removed in vacuo. The crude product was purified by chromatography on silica gel (n-hexane / EA). MS (m / z): 306.6 [M + H ⁺ ]

F. 2-Methyl-propane-2-sulfinic acid [1- (5-Bromo-3-fluoro-pyridin-2-yl) -ethyl] -amide 2-Methyl-propane-2-sulfinic acid in DCM (570 mL) 5 -A solution of bromo-3-fluoro-pyridin-2-ylmethyleneamide (17.5 g, 0.06 mol) in methyl magnesium chloride solution (29.9 ml, 0.09 mol, 22 in THF) at -55 ° C. %) Was added slowly. After stirring at −55 ° C. for 20 minutes, the reaction mixture was quenched with brine. Celite was added and the suspension was filtered. The layers were separated and the aqueous layer was extracted with DCM. The combined organic layers were dried over sodium sulfate and the solvent removed in vacuo. . The crude product was purified by chromatography on silica gel (n-hexane / EA). MS (m / z): 322.6 [M + H ⁺ ]

G. 5-Fluoro-6- [1- (2-methyl-propane-2-sulfinylamino) -ethyl] -nicotinic acid methyl ester 2-methyl-propane-2-sulfinic acid in anhydrous DMSO (10 mL) [1- ( 5-Bromo-3-fluoro-pyridin-2-yl) -ethyl] -amide (1.40 g, 4.33 mmol), 1,4-bis- (diphenylphosphino) ferrocene (0.53 g, 0.96) Mmol) and Pd (OAc) ₂ (0.21 g, 0.91 mmol) into a stirred suspension of triethylamine (3.6 mL, 26.0 mmol) and anhydrous methanol (4 mL) under an atmosphere of nitrogen. Was added. After stirring at 70 ° C. for 14 hours under an atmosphere of carbon monoxide, the mixture was concentrated in vacuo and the residue was partitioned between EA (200 mL) and saturated NaHCO ₃ solution (200 mL). After extraction of the aqueous layer with EA (2 × 200 mL), the combined organic layers were dried over Na ₂ SO ₄ , filtered and concentrated in vacuo. The residue was purified by flash chromatography on silica gel (eluting with n-hexane / EA 19: 1) to give the title compound. MS (m / z): 302.7 [M + H ⁺ ]

H. 6-((1R) -1- (1,1-dimethylethylsulfinamido) ethyl) -5-fluoronicotinic acid 5-fluoro-6--6 in MeOH (1.5 mL) and H ₂ O (0.5 mL) To a solution of [1- (2-methyl-propane-2-sulfinylamino) -ethyl] -nicotinic acid methyl ester (910 mg, 3.01 mmol) was added LiOH (215 mg, 9.03 mmol). The solution was stirred overnight and then concentrated. The residue was dissolved in 15% MeOH / CH ₂ Cl ₂ and filtered through a short silica column to remove inorganic salts. Crude 6-((1R) -1- (1,1-dimethylethylsulfinamido) ethyl) -5-fluoronicotinic acid was determined to be of sufficient purity for the next reaction. MS (m / z): 288.7 [M + H ^<+ >]

I. N-((R) -1- (5-((2R, 5S) -2,5-dimethylpyrrolidin-1-carbonyl) -3-fluoropyridin-2-yl) ethyl) -2-methylpropane-2- Sulfinamide (2R, 5S) -2,5-dimethyltetrahydro-1H-1-pyrrolidinium chloride (Example 4C) (35.3 mg, 0.26 mmol), HATU (65 0.9 mg, 0.17 mmol) and DIPEA (151 μL, 0.87 mmol) to a stirred solution of 6-((1R) -1- (1,1-dimethylethylsulfinamido) ethyl) -5-fluoro Nicotinic acid (50.0 mg, 0.17 mmol) was added. After stirring overnight, the reaction mixture was concentrated in vacuo and the residue was partitioned between EA (30 mL) and saturated NaHCO ₃ solution (50 mL). After extraction of the aqueous layer with EA (2 × 30 mL), the combined organic layers were dried over Na ₂ SO ₄ , filtered and concentrated in vacuo to give the crude title compound, which was flash chromatographed on silica gel. (Elution with n-hexane / EA 5: 1). MS (m / z): 369.8 [M + H ^<+ >]

J. et al. (6-((R) -1-aminoethyl) -5-fluoropyridin-3-yl) ((2R, 5S) -2,5-dimethylpyrrolidin-1-yl) methanone hydrochloride N-((R) -1- (5-((2R, 5S) -2,5-dimethylpyrrolidin-1-carbonyl) -3-fluoropyridin-2-yl) ethyl) -2-methylpropane-2-sulfinamide (50 mg, 0 To a solution of HCl in MeOH (3N, 1 mL). The reaction mixture is stirred at room temperature for 2 hours and then concentrated to dryness. A purification step is not necessary and the isolated title compound is used in the next reaction. MS (m / z): 265.9 [M + H ⁺ ]

K. 2-((R) -1- (5-((2R, 5S) -2,5-dimethylpyrrolidin-1-carbonyl) -3-fluoropyridin-2-yl) ethylcarbamoyl) -2-ethylhydrazinecarboxylic acid tert-Butyl tert-Butyl 2- (chlorocarbonyl) -2-ethylhydrazinecarboxylate was synthesized according to the literature (Bailey et al., J. Med. Chem. 47, 2004, 3788-3799).

Tert-butyl 2- (chlorocarbonyl) -2-ethylhydrazinecarboxylate (46 mg, 0.21 mmol) was dissolved in THF (2 mL), treated with DIPEA (361 μL, 2.07 mmol) and brought to 0 ° C. After cooling, (6-((R) -1-aminoethyl) -5-fluoropyridin-3-yl) ((2R, 5S) -2,5-dimethylpyrrolidine-1- in THF (2 mL) Yl) methanone A solution of hydrochloride (25 mg, 0.08 mmol) and DIPEA (32 μL, 0.18 mmol) was added in one portion. The reaction was kept at 0 ° C. (1 hour) and then stirred at room temperature for 14 hours. The mixture is poured into EA and washed successively with HCl (1N), saturated NaHCO ₃ and brine, then dried (MgSO ₄ ), filtered,
Concentrated. The product was flash chromatographed on silica gel (n-hexane / EA
Elution with 3: 1). MS (m / z): 451.9 [M + H ⁺ ]

L. N-((R) -1- (5-((2R, 5S) -2,5-dimethylpyrrolidin-1-carbonyl) -3-fluoropyridin-2-yl) ethyl) -1-ethyl-2- ( 3-Fluoro-5- (trifluoromethyl) benzoyl) hydrazinecarboxamide 2-((R) -1- (5-((2R, 5S) -2,5-dimethylpyrrolidine-1-carbonyl) -3-fluoro To a solution of tert-butyl pyridin-2-yl) ethylcarbamoyl) -2-ethylhydrazinecarboxylate (25 mg, 0.06 mmol) is added a solution of HCl in MeOH (3N, 1 mL). The reaction mixture is stirred at room temperature for 2 hours and then concentrated to dryness. A purification step is not necessary and the isolated hydrochloride salt is used in the next reaction.

Hydrochloride (20 mg, 0.05 mmol) was dissolved in DMF (0.5 mL) and DIPEA (90 μL, 0.52 mmol) and 3-fluoro-5- (trifluoromethyl) benzoyl chloride (10 mg, .0. 08 mmol) and stir at room temperature for 2 hours. The mixture was then concentrated in vacuo and the residue was purified by reverse phase HPLC using a gradient of ACN in water containing 0.1% TFA to give the title compound. MS (m / z): 541.8 [M + H ^<+ >]

(R) -N- (1- (2-chloro-4- (diethylcarbamoyl) phenyl) ethyl) -1-ethyl-2- (3-fluoro-5- (trifluoromethyl) benzoyl) hydrazinecarboxamide (compound 256)

A. 4-((tert-Butylsulfinylimino) methyl) -3-chlorobenzoic acid methyl The synthesis of 3-chloro-4-formyl-benzoic acid methyl ester is described in Example 15B.

(S)-(−)-2-methyl-2-propanesulfinamide (1.26 g, 10.4 mol), magnesium sulfate (12.5 g, 104 mmol), PPTS (0.52 g, 2.08 mol) Was added to a solution of 3-chloro-4-formyl-benzoic acid methyl ester (2.07 g, 10.4 mmol) in DCM (52 mL). After stirring overnight, the reaction was filtered over celite and the solvent removed in vacuo. The crude product was purified by chromatography on silica gel (n-hexane / EA). MS (m / z): 301.7 [M + H ⁺ ]

B. 3-Chloro-4-((1R) -1- (1,1-dimethylethylsulfinamido) ethyl) methyl benzoate 4-((tert-butylsulfinylimino) methyl) -3-chloro in DCM (30 mL) To a solution of methyl benzoate (1.73 g, 5.73 mmol), methylmagnesium chloride solution (3.4 ml, 9.75 mmol, 22% in THF) was slowly added at -55 ° C. After stirring at −55 ° C. for 1 hour, the reaction mixture was quenched with brine. Celite was added and the suspension was filtered. The layers were separated and the aqueous layer was extracted with DCM. The combined organic layers were dried over sodium sulfate and the solvent removed in vacuo. . The crude product was purified by chromatography on silica gel (n-hexane / EA). MS (m / z): 317.8 [M + H ^<+ >]

C. 4- (1-Aminoethyl) -3-chlorobenzoic acid (R) -methyl hydrochloride 3-Chloro-4-((1R) -1- (1,1-dimethylethylsulfinamido) ethyl) methyl benzoate ( To a solution of 199 mg, 0.63 mmol) is added a solution of HCl in MeOH (3N, 10 mL). The reaction mixture is stirred at room temperature for 1 hour and then concentrated to dryness. A purification step is not necessary and the isolated title compound is used in the next reaction. MS (m / z): 213.8 [M + H ^<+ >]

D. 2- (1- (2-Chloro-4- (methoxycarbonyl) phenyl) ethylcarbamoyl) -2-ethylhydrazinecarboxylic acid (R) -tert-butyl 2- (chlorocarbonyl) -2-ethylhydrazinecarboxylic acid tert- Butyl was synthesized according to the literature (Bailey et al., J. Med. Chem. 47, 2004, 3788-3799).

Tert-butyl 2- (chlorocarbonyl) -2-ethylhydrazinecarboxylate (417 mg, 1.87 mmol) was dissolved in THF (2 mL) and treated with DIPEA (718 μL, 4.11 mmol)) at 0 ° C. After cooling to 4- (1-aminoethyl) -3-chlorobenzoic acid (R) -methyl hydrochloride (79 mg, 0.32 mmol) and DIPEA (121 μL, 0.69 mmol) in THF (2 mL). ) Was added all at once. The reaction was kept at 0 ° C. (1 hour) and then stirred at room temperature for 14 hours. The mixture was poured into EA and washed successively with HCl (1N), saturated NaHCO ₃ and brine, then dried (MgSO ₄ ), filtered and concentrated. The product was purified by flash chromatography on silica gel (eluting with n-hexane / EA). MS (m / z): 399.7 [M + H ⁺ ]

E. 3-chloro-4- (1- (1-ethyl-2- (3-fluoro-5- (trifluoromethyl) benzoyl) hydrazinecarboxamido) ethyl) benzoic acid (R) -methyl 2- (1- (2 -Chloro-4- (methoxycarbonyl) phenyl) ethylcarbamoyl) -2-ethylhydrazinecarboxylic acid (R) -tert-butyl (75 mg, 0.19 mmol) was added a solution of HCl in MeOH (3N, 2 mL). Add. The reaction mixture is stirred at room temperature for 14 hours and then concentrated to dryness. A purification step is not necessary and the isolated title compound is used in the next reaction.

Hydrochloride (63 mg, 0.19 mmol) was dissolved in DMF (1.0 mL) and DIPEA (163 μL, 0.94 mmol) and 3-fluoro-5- (trifluoromethyl) benzoyl chloride (47 mg,. 21 mmol) and stirred at room temperature for 2 hours. The mixture was then concentrated in vacuo and the residue was purified by chromatography on silica gel (n-hexane / EA). MS (m / z): 489.6 [M + H ⁺ ]

F. (R) -N- (1- (2-chloro-4- (diethylcarbamoyl) phenyl) ethyl) -1-ethyl-2- (3-fluoro-5- (trifluoromethyl) benzoyl) hydrazinecarboxamide dioxane ( 3-chloro-4- (1- (1-ethyl-2- (3-fluoro-5- (trifluoromethyl) benzoyl) hydrazinecarboxamido) ethyl) benzoic acid (R) -methyl (92 mg, 2 mL) To a solution of 0.19 mmol) was added a 2N solution of LiOH in water (1 mL) and the reaction was stirred for 3 hours. The organic solvent was then removed in vacuo, the aqueous layer was extracted twice with EA, the combined organic layers were dried over sodium sulfate, and the solvent was removed in vacuo. The crude product was used in the next step without further purification.

(R) -3-Chloro-4- (1- (1-ethyl-2- (3-fluoro-5- (trifluoromethyl) benzoyl) hydrazinecarboxamido) ethyl) benzoic acid (27.0 mg, 0.06 Mmol) was added to a stirred solution of diethylamine (44.0 mg, 0.60 mmol) and HATU (22.9 mg, 0.06 mmol) in DMF (1.5 mL). After stirring for 2 hours, the reaction mixture was concentrated in vacuo and the residue was purified by reverse phase HPLC using a gradient of ACN in water containing 0.1% TFA to give the title compound. MS (m / z): 530.9 [M + H ^<+ >]

N-((R) -1- (3-chloro-5-((2R, 5S) -2,5-dimethylpyrrolidin-1-carbonyl) pyridin-2-yl) ethyl) -1-ethyl-2- ( Synthesis of 3-fluoro-5- (trifluoromethyl) benzoyl) hydrazinecarboxamide (Compound 263)

A. Dimethyl 3-chloropyridine-2,5-dicarboxylate Dimethyl 3-chloropyridine-2,5-dicarboxylate was synthesized according to the literature (Hendrickson et al., Org. Lett. 2004, 3-5).

Pyridine-2,5-dicarboxylic acid (10.0 g, 60.0 mmol) was suspended in 300 mL of 0.2% (w / w) Na ₂ WO ₄ aqueous solution (0.6 g). To this solution was added H ₂ O _{2 in} water (30% w / w, 34 mL, 315 mmol). The resulting mixture was stirred at 80-85 ° C. for 10 hours and heated. The resulting solid was collected by filtration and washed with cold water. Drying the material under high vacuum overnight gave pyridine-2,5-dicarboxylic acid N-oxide, which was used in the next step without further purification.

To a solution of thionyl chloride (17.4 mL, 240 mmol) in DCM (500 mL) was added DMF (5 mL) at 0 ° C. Pyridine-2,5-dicarboxylic acid N-oxide from the first stage was added in portions to this mixture. The resulting mixture was heated at 65 ° C. for 2 hours. The reaction was cooled to room temperature and then placed in an ice-bath. The reaction was quenched using methanol (100 mL) slowly at 0 ° C. The solvent was removed in vacuo and the crude product was partitioned between DCM (200 mL) and aqueous NaHCO ₃ (saturated 100 mL). The organic layer was separated and the aqueous layer was extracted with additional DCM (2 × 200 mL). The combined organic layers were dried (Na ₂ SO ₄ ), filtered and the solvent removed in vacuo. Purification of the crude oil through a short plug of silica gel with hexane / ethyl acetate (4: 1) gave dimethyl 3-chloropyridine-2,5-dicarboxylate. GC / MS (m / z): 229.0

B. To a solution of dimethyl 3-chloropyridine-2,5-dicarboxylate (1.0 g, 4.4 mmol) in methyl 5-chloro-6- (hydroxymethyl) nicotinate THF (10 mL) and MeOH (20 mL) was added. Powdered calcium chloride (3.9 g, 35 mmol) was added. The suspension was cooled to 0 ° C. and sodium borohydride (416 mg, 11.0 mmol) was added in portions. The resulting mixture was stirred at 0 ° C. for 3 hours, poured into ice-water (200 mL) and extracted with DCM (2 × 100 mL). The combined organic layers are dried (Na ₂ SO ₄ ), filtered and the solvent removed in vacuo to give methyl 5-chloro-6- (hydroxymethyl) nicotinate, which is used in the next step without further purification. Used in. GC / MS (m / z): 201.0

C. Methyl 5-chloro-6-formylnicotinate To a solution of methyl 5-chloro-6- (hydroxymethyl) nicotinate (3.2 g, 15.8 mmol) in DCM (20 mL) was added Dess-Martin periodinane (10. 1 g, 23.7 mmol) was added. The solution was stirred for 1 h, diluted with DCM and filtered over a pad of celite. After evaporation of the solvent in vacuo, the crude material was purified by flash chromatography (hexane / ethyl acetate 3: 1) to give the title compound. GC / MS (m / z): 201.0

D. 6-((tert-Butylsulfinylimino) methyl) -5-chloronicotinic acid (R) -methyl (R)-(−)-2-methyl-2-propanesulfinamide (0.73 g) in DCM (25 mL) , 6.0 mmol) in a solution of methyl 5-chloro-6-formylnicotinate (1.2 g, 6.0 mmol), pyridinium p-toluenesulfonate (0.15 mg, 0.6 mmol), magnesium sulfate (7.2 g, 60 mmol) and freshly ground cesium carbonate (1.9 g, 6.0 mmol) were added. The reaction was stirred at room temperature for 6 hours. The mixture was filtered through celite, washed with DCM and concentrated in vacuo. The resulting residue was subjected to silica gel chromatography (0-10% ethyl acetate in hexanes) to give the title compound. MS (m / z): 302.6 [M + H ⁺ ]

E. 5-chloro-6-((R) -1-((R) -1,1-dimethylethylsulfinamido) ethyl) methyl nicotinate 6-((tert-butylsulfinylimino) methyl) in DCM (30 mL) To a stirred solution of (5-chloronicotinic acid (R) -methyl (1.2 g, 4.0 mmol) at −60 ° C. methyl magnesium chloride (22% in THF, 2.1 mL, 6.0 mmol). ) Was added dropwise. After stirring at −60 ° C. for 30 minutes, saturated ammonium chloride solution (40 mL) was added. The aqueous layer was extracted with DCM (4 × 25 mL) and the organic layer was dried over Na ₂ SO ₄ , filtered and concentrated in vacuo. The residue was purified by flash chromatography on silica gel (eluting with hexane / EA 1: 1) to give the title compound. MS (m / z): 318.8 [M + H ⁺ ]

F. 6- (1-Aminoethyl) -5-chloronicotinic acid (R) -methyl hydrochloride 5-chloro-6-((R) -1-((R) -1,1-dimethylethylsulfinamido) ethyl) A solution of methyl nicotinate (230 mg, 0.72 mmol) and HCl (3N solution in MeOH, 2 mL) was stirred at room temperature for 1.5 hours. The solvent was removed in vacuo and the residue was recrystallized from diethyl ether to give the title compound as the HCl salt. MS (m / z): 214.8 [M + H ⁺ ]

G. 6- (1- (2- (tert-butoxycarbonyl) -1-ethylhydrazinecarboxamido) ethyl) -5-chloronicotinic acid (R) -methyl according to literature 2- (chlorocarbonyl) -2-ethylhydrazinecarboxylic acid Tert-butyl was synthesized (Bailey et al., J. Med. Chem. 47, 2004, 3788-3799).

Tert-butyl 2- (chlorocarbonyl) -2-ethylhydrazinecarboxylate (443 mg, 1.99 mmol) was dissolved in THF (2.5 mL) and treated with DIPEA (705 μL, 3.98 mmol) After cooling to ° C., 6- (1-aminoethyl) -5-chloronicotinic acid (R) -methyl hydrochloride (100 mg, 0.40 mmol) and DIPEA (155 μL, 0) in THF (2.5 mL). .88 mmol) solution was added in one portion. The reaction was kept at 0 ° C. (1 hour) and then stirred at room temperature for 14 hours. The mixture was poured into EA and washed successively with HCl (1N), saturated NaHCO ₃ and brine, then dried (MgSO ₄ ), filtered and concentrated. The product was purified by flash chromatography on silica gel (eluting with hexane / EA). MS (m / z): 400.8 [M + H ⁺ ]

H. (R) -6- (1- (2- (tert-butoxycarbonyl) -1-ethylhydrazinecarboxamido) ethyl) -5-chloronicotinic acid 6- (1- (2- (tert-butoxycarbonyl) -1 -Ethylhydrazinecarboxamido) ethyl) -5-chloronicotinic acid (R) -methyl (220 mg, 0.55 mmol) was dissolved in ACN (5 mL) and treated with LiOH (1M solution in water, 3.5 mL). And stirred at room temperature for 13 hours. The mixture was concentrated in vacuo and EA (150 mL) was added. The pH of the aqueous layer was adjusted to 4 by addition of 1N HCl solution, and the aqueous layer was extracted with EA (3x) and DCM / iPrOH (4: 1, 3x). The combined organic layers were dried over Na ₂ SO ₄ , filtered and concentrated in vacuo. The resulting title compound was used in the next reaction without purification. MS (m / z): 386.8 [M + H ^<+ >]

I. 2-((R) -1- (3-Chloro-5-((2R, 5S) -2,5-dimethylpyrrolidin-1-carbonyl) pyridin-2-yl) ethylcarbamoyl) -2-ethylhydrazinecarboxylic acid tert-Butyl (R) -6- (1- (2- (tert-Butoxycarbonyl) -1-ethylhydrazinecarboxamido) ethyl) -5-chloronicotinic acid (150 mg, 0. 1) in DCM (2.5 mL). 39 mmol), WSC (149 mg, 0.78 mmol), HOBt (105 mg, 0.78 mmol) and DIPEA (330 μL, 1.94 mmol) into a stirred solution of (2R, 5S) -2,5- Dimethylpyrrolidine hydrochloride (105 mg, 0.78 mmol) was added. After stirring at room temperature overnight, the solvent was removed in vacuo and the crude product was partitioned between EE and aqueous Na ₂ CO ₃ solution. The organic layer was separated and the aqueous layer was extracted with additional EA (2x). The combined organic layers were dried (Na ₂ SO ₄ ), filtered and the solvent removed in vacuo. The resulting title compound was used in the next reaction without purification. MS (m / z): 467.9 [M + H ^<+ >]

J. et al. N-((R) -1- (3-chloro-5-((2R, 5S) -2,5-dimethylpyrrolidin-1-carbonyl) pyridin-2-yl) ethyl) -1-ethyl-2- ( 3-Fluoro-5- (trifluoromethyl) benzoyl) hydrazinecarboxamide 2-((R) -1- (3-chloro-5-((2R, 5S) -2,5-dimethylpyrrolidine-1-carbonyl) To a solution of tert-butyl pyridin-2-yl) ethylcarbamoyl) -2-ethylhydrazinecarboxylate (180 mg, 0.39 mmol) is added a solution of HCl in MeOH (3N, 20 mL). The reaction mixture is stirred at room temperature for 13 hours and then concentrated to dryness. A purification step is not necessary and the isolated hydrochloride salt is used in the next reaction.

N-((R) -1- (3-chloro-5-((2R, 5S) -2,5-dimethylpyrrolidin-1-carbonyl) pyridin-2-yl) ethyl) -1-ethyl-2- ( 3-Fluoro-5- (trifluoromethyl) benzoyl) hydrazinecarboxamide hydrochloride (8.0 mg, 0.02 mmol) was added to 3-fluoro-5- (trifluoromethyl) benzoic acid in DMF (0.2 mL). Stirred solution of acid (9.1 mg, 0.04 mmol), WSC (8.3 mg, 0.04 mmol), HOBt (5.9 mg, 0.04 mmol) and DIPEA (19 μL, 0.11 mmol) Added to. After stirring at room temperature overnight, the residue was purified by reverse phase HPLC using a gradient of ACN in water containing 0.1% TFA to give the title compound. MS (m / z): 557.9 [M + H ^<+ >]

(R) -5-chloro-N, N-diethyl-6- (1- (1-ethyl-2- (3-fluoro-5- (trifluoromethyl) benzoyl) hydrazinecarboxamido) ethyl) nicotinamide (compound 265)

A. 2- (1- (3-Chloro-5- (diethylcarbamoyl) pyridin-2-yl) ethylcarbamoyl) -2-ethylhydrazinecarboxylic acid (R) -tert-butyl (R) in DCM (1.5 mL) -6- (1- (2- (tert-butoxycarbonyl) -1-ethylhydrazinecarboxamido) ethyl) -5-chloronicotinic acid (82 mg, 0.21 mmol), WSC (82 mg, 0.42 mmol), To a solution of HOBt (57 mg, 0.42 mmol) and DIPEA (180 μL, 1.06 mmol) was added diethylamine (31 mg, 0.42 mmol). After stirring at room temperature overnight, the solvent was removed in vacuo and the crude product was partitioned between EE and aqueous Na ₂ CO ₃ solution. The organic layer was separated and the aqueous layer was extracted with additional EA (2x). The combined organic layers were dried (Na ₂ SO ₄ ), filtered and the solvent removed in vacuo. The resulting title compound was used in the next reaction without purification. MS (m / z): 441.9 [M + H ^<+ >]

B. N-((R) -1- (3-chloro-5-((2R, 5S) -2,5-dimethylpyrrolidin-1-carbonyl) pyridin-2-yl) ethyl) -1-ethyl-2- ( 3-Fluoro-5- (trifluoromethyl) benzoyl) hydrazinecarboxamide 2- (1- (3-chloro-5- (diethylcarbamoyl) pyridin-2-yl) ethylcarbamoyl) -2-ethylhydrazinecarboxylic acid (R ) -Tert-Butyl (93 mg, 0.21 mmol) is added a solution of HCl in MeOH (3N, 15 mL). The reaction mixture is stirred at room temperature for 13 hours and then concentrated to dryness. A purification step is not necessary and the isolated hydrochloride salt is used in the next reaction.

(R) -5-chloro-N, N-diethyl-6- (1- (1-ethylhydrazinecarboxamido) ethyl) nitrinamide hydrochloride (8.3 mg, 0.02 mmol) was added to DMF (0.2 mL). 3-fluoro-5- (trifluoromethyl) benzoic acid (10 mg, 0
. 05 mmol), WSC (9.3 mg, 0.05 mmol), HOBt (6.6 mg, 0.05 mmol) and DIPEA (22 μL, 0.12 mmol) were added to a stirred solution. After stirring at room temperature overnight, the residue was purified by reverse phase HPLC using a gradient of ACN in water containing 0.1% TFA to give the title compound. MS (m / z): 531.8 [M + H ^<+ >]

(R) -N- (1- (4-Diethylcarbamoyl) -2-fluorophenyl) ethyl) -1-ethyl-2- (3-fluoro-5- (trifluoromethyl) benzoyl) hydrazinecarboxamide (Compound 221 )

A. (R) -4- (1-(((9H-Fluoren-9-yl) methoxy) carbonylamino) ethyl) -3-fluorobenzoic acid (R) -4- (1-amino-ethyl) -3-fluoro -Methyl benzoate (Example 10C) (100 mg, 0.43 mmol) was stirred in 6N HCl solution (10 mL) at reflux overnight. The solvent was reduced in vacuo to give (R) -4- (1-aminoethyl) -3-fluorobenzoic acid hydrochloride. To the crude product (94 mg, 0.43 mmol) dissolved in DCM (2 mL) was added chlorotrimethylsilane (135 μL, 1.07 mmol). After stirring for several minutes at reflux, (9H-fluoren-9-yl) methylcarbonochloridate and triethylamine (230 mL, 1.7 mmol) were added at room temperature and the solution was stirred overnight. The residue was concentrated in vacuo and saturated Na ₂ CO ₃ solution (50 mL) was added. After acidification of the aqueous solution with 2N HCl (pH = 3), the aqueous layer was extracted with EA (2 × 30 mL) and the combined organic layers were dried over Na ₂ SO ₄ , filtered and concentrated in vacuo. The crude title compound was obtained, which was purified by flash chromatography on silica gel (eluting with DCM / methanol 2: 1). MS (m / z): 405.8 [M + H ^<+ >]

B. Resin-bonded (R) -4- (1-aminoethyl) -3-fluorobenzoic acid (R) -4- (1-(((9H-fluoren-9-yl) methoxy) carbonylamino) ethyl) -3 -Fluorobenzoic acid (324 mg, 0.8 mmol) and DIPEA (332 μL, 1.9 mmol) were dissolved in DCM (8 mL). The solution was added to 2-chlorotrityl chloride resin (loading: 1.4 mmol / g, 1.09 g, 1.5 mmol) swollen in DCM (8 mL) and the mixture was shaken at room temperature for 30 min. The mixture was quenched with methanol (800 μL). The resin was filtered and washed with DCM / methanol / DIPEA (16: 3: 3, 1 × 10 mL) and DMF (3 × 10 mL). Piperidine (25% solution in DMF, 10 mL) was added and the mixture was shaken for 20 minutes. The resin was filtered, washed with DMF (6 × 10 mL) and dried in vacuo to give the title compound at the expected loading of 45% (0.65 mmol / g).

C. 2-Ethylhydrazinecarboxylic acid (9H-fluoren-9-yl) methyl A solution of hydrazinecarboxylic acid (9H-fluoren-9-yl) methyl (500 mg, 1.97 mmol) in ethanol (9.8 mL) was added to acetaldehyde. (86.7 mg, 1.97 mmol) was added and the mixture was refluxed overnight. To this solution was added acetic acid (124 μL, 2.16 mmol) and sodium cyanotrihydroborate (136 mg, 2.16 mmol). After stirring at room temperature overnight, the solvent was reduced in vacuo and the residue was partitioned between EA (30 mL) and saturated NaCl solution (50 mL). After extraction of the aqueous layer with EA (2 × 30 mL), the combined organic layers were dried over Na ₂ SO ₄ , filtered and concentrated in vacuo to give the crude title compound, which was flash chromatographed on silica gel. Purified by chromatography (elution with n-hexane / EA 1: 1). MS (m / z): 282.7 [M + H ^<+ >]

D. (R) -4- (1- (1-Ethyl-2- (3-fluoro-5- (trifluoromethyl) benzoyl) hydrazinecarboxamido) ethyl) -3-fluorobenzoic acid in DCM (4.4 mL) To a stirred solution of methyl 2-ethylhydrazinecarboxylate (9H-fluoren-9-yl) (125 mg, 0.44 mmol) was added phosgene (20% solution in toluene, 0.463 ml, 0.88 mmol). It was dripped at 0 ° C. The mixture was stirred at 0 ° C. for 30 minutes and then concentrated in vacuo to give 2- (chlorocarbonyl) -2-ethylhydrazinecarboxylic acid (9H-fluoren-9-yl) methyl. The crude product (68.5 mg, 0.207 mmol) was dissolved in DCM (1 mL) and swollen in DCM (5 mL) and collidine (136 μL, 1.035 mmol). Resin-bond (R) -4 To a suspension of-(1-amino-ethyl) -3-fluorobenzoic acid (106 mg, 0.069 mmol). The mixture was shaken for 90 minutes at room temperature. The resin was filtered and washed with DMF (4 × 2 mL), methanol (4 × 2 mL) and DCM (4 × 2 mL). Test cleavage with TFA yields the correct product (R) -4- (1- (2-(((9H-fluoren-9-yl) methoxy) carbonyl) -1-ethylhydrazinecarboxamido) ethyl) -3- Fluorobenzoic acid was indicated. MS (m / z): 491.7 [M + H ^<+ >]. Piperidine (20% solution in DMF, 4 mL) is added and the mixture is shaken for 30 minutes to give resin-bound (R) -4- (1- (1-ethylhydrazinecarboxamido) ethyl) -3-fluorobenzoic acid. Acid was given. The resin was filtered, washed with DMF (4 × 2 mL), methanol (4 × 2 mL) and DCM (4 × 2 mL) and 3-fluoro-5-trifluoromethyl-benzoic acid (17 in DCM / DMF 2: 1 (1.5 ml). .2 mg, 0.083 mmol), HATU (31.5 mg, 0.083 mmol) and collidine (45.4 μL, 0.345 mmol) were added. The mixture was shaken for 4 hours at room temperature. The resin was filtered and washed with DMF (4 × 2 mL), methanol (4 × 2 mL) and DCM (4 × 2 mL) and suspended in DCM (1 mL) and TFA (1 mL). The suspension was shaken for 20 minutes at room temperature. The resin was filtered and washed with DCM (2 × 2 mL). The filtrate was concentrated in vacuo. The residue was purified by reverse phase HPLC using a gradient of ACN in water containing 0.1% TFA to give the title compound. MS (m / z): 459.6 [M + H ⁺ ]

E. (R) -N- (1- (4-Diethylcarbamoyl) -2-fluorophenyl) ethyl) -1-ethyl-2- (3-fluoro-5- (trifluoromethyl) benzoyl) hydrazinecarboxamide (R) -4- (1- (1-ethyl-2- (3-fluoro-5- (trifluoromethyl) benzoyl) hydrazinecarboxamido) ethyl) -3-fluorobenzoic acid (12 mg, 0.028 mmol) was added to DMF. (1.5 mL) was added to a stirred solution of diethylamine (5.7 μL, 0.055 mmol), HATU (10.6 mg, 0.028 mmol) and DIPEA (23.4 μL, 0.14 mmol). . After stirring overnight, the reaction mixture was concentrated in vacuo and the residue was partitioned between EA (30 mL) and saturated NaHCO ₃ solution (50 mL). After extraction of the aqueous layer with EA (2 × 30 mL), the combined organic layers were dried over Na ₂ SO ₄ , filtered and concentrated in vacuo to give the crude title compound in water containing 0.1% TFA. It was purified by reverse phase HPLC using a gradient of ACN to give the title compound. MS (m / z): 514.8 [M + H ^<+ >]

Biological evaluation:

Radioligand binding assay Assessing the affinity of selected compounds that bind to different types of BK Bl receptors HEK923 can be induced to express human, rabbit, mouse, dog, pig or rat Bl receptors by tetracycline Radioligand binding assays were performed using cells.

  Using gene design and codon usage optimized for stable expression in human cells, the genes for human, rat, mouse, rabbit, dog and pig BK B1R are synthesized synthetically (GENEART, Regensburg) was formed. Use of the Flp-In system from Invitrogen with the host cell line Flp-In T-REx HEK293 (human embryonic kidney) together with pcDNA5 / FRT / TO-vector (for stable but tetracycline-inducible expression) 1, all receptor genes were stably and isogenically expressed by insertion into the same position in the genome. The pcDNA5 / FRT / TO-vector avoids all B1R subsequences in order to avoid problems with the potential for constitutive activity of these receptors that could prevent the growth and selection of stably expressing clones. Especially used for type.

  For all receptor types, each HEK293 cell harboring each transgene was cultured in DMEM high glucose medium supplemented with 10% FCS. At 80% confluence, cells were harvested from the culture flask by trypsinization and resuspended in DMEM 10% FCS containing 5 μg / mL tetracycline. Cells were seeded at approximately 80.000 cells per well in poly-lysine coated flat bottom 96 well plates (TPP) and used in binding assays after overnight incubation at 37 ° C.

Cell plates were washed twice with ice cold PBS and kept on ice throughout the assay. Test compounds were assayed at 10 concentrations diluted in half-log steps in duplicate wells. Assay binding buffer (40 mM PIPES, 109 mM NaCl, 5 mM KCl, 0.1% glucose) containing the protease inhibitors Captopril (100 μM), 1,10-phenanthroline (20 μM) and bacitracin (500 μM), Compounds were diluted in 0.05% BSA, 2 mM CaCl ₂ , 1 mM MgCl ₂ , 60 mM NaOH; pH 7.4). 1 nM 3H-DAKD (Perkin Elmer) and antagonist compound dilutions are prepared in dilution plates, which are standards for assessing total binding (1 nM 3H-DAKD) and standards for assessing non-specific binding (1 nM
3H DAKD + 10 mM DAKD). 100 μL from the dilution plate was added to the cell plate and incubated for 90 minutes on ice. The supernatant was then aspirated and the plate was washed 4 times with ice-cold PBS, followed by the addition of 200 μL of dissociation buffer (0.5M NaCl, 0.2M acetic acid) for 10 minutes on ice.

Transfer the supernatant into a 6 mL scintillation bin (Sarstedt) containing 2 mL scintillation solution (Ultima Gold, Perkin-Elmer), mix well, followed by Packard Topcount Scintillation.
Measured at the counter. Specific binding to B1R was defined as the difference between total binding and non-specific binding wells. Total specific binding was set at 100% binding and 0% inhibition. Specific counts in the presence of compound were plotted against compound concentration (log M). IC ₅₀ values were calculated (Xlfit, IDBS ID Business Solutions Ltd) by fitting a 4-parameter logic function to the concentration-response data using non-linear regression. The compounds of the invention have an affinity for B1R of less than 5 μM, as shown by the results in the above assay.

Functional Assay for BK B1 Antagonists The potency and efficacy of compounds in the present invention that antagonize BK B1R were determined in a cell-based fluorescent calcium-mobilization assay. The assay measures the ability of a test compound to inhibit the BK B1R agonist-induced increase in intracellular free calcium in various cell lines. Endogenous B1R expression was induced by IL-1 beta pretreatment of human embryonic lung fibroblasts IMR-90. Several sub-human species (rat, mouse, rabbit, dog and pig) B1R were recombinantly expressed following tetracycline induction in HEK 293 cells harboring the respective transgene.

  Cells loaded with calcium-indicator were preincubated in the absence or presence of various concentrations of test compound followed by stimulation with selective B1R agonist peptides. B1R agonist-induced calcium mobilization was monitored using a FlexStation fluorescence imaging plate reader platform.

IMR-90 human embryonic lung fibroblasts (ATCC # CCL 186) were cultured in DMEM high (4.5 g / l) glucose supplemented with 15% FCS and 4 mM glutamine. After 5 days of culture, confluent cells were harvested by trypsinization and seeded at 40.000 cells per well in black wall / clear bottom 96-well plates (Costar # 3603). After overnight incubation (approximately 12-16 hours), cells were treated with 1.36 ng / mL human recombinant IL-1β in DMEM 15% FBS for 4 hours at 37 ° C. to upregulate B1R. Induced. Thereafter, the cells were washed twice with Hank's balanced salt solution / 20 mM HEPES buffer, followed by the presence of the anion transport inhibitor probenecid at 2.5 mM. Below, the fluorescent calcium indicator dye no-wash Calcium 3 assay kit (Molecular Devices) was loaded for 1 hour at 37 ° C. Test compounds were assayed at 7 concentrations in duplicate wells. Compound loaded plates contained test compounds or standards at a concentration 5 times the final concentration in 5% DMSO. Test compound was added in 5 μL followed by a 10 minute equilibration step at 37 ° C. in an incubator. The plates were then placed in a FlexStation II apparatus (Molecular Devices) set at 37 ° C. While continuously monitoring the Ca-dependent increase in fluorescence, 50 μL of B1R agonist desArgKallidin (DAKD, Bachem) was added at FlexStation II. The fluorescence peak height was calculated by subtracting the baseline average from the maximum peak height. Peak height without added antagonist was defined as 100% response and 0% inhibition, and peak height was plotted as a function of test compound concentration. The relative fluorescence peak height was used to calculate the degree of inhibition of the B1R agonist response by the test compound. IC ₅₀ values were calculated by fitting a 4-parameter logic function to concentration-response data using non-linear regression (Xlfit, IDBS ID Business So
lutions Ltd). The compounds of the invention have an affinity for B1R of less than 5 μM, as shown by the results in the above assay.

The most preferred compounds of formula (I) exhibited IC ₅₀ values of less than 50 nanomolar in this assay.

  The features of the invention disclosed in the description, claims and / or drawings can be materials for implementing the invention in its various forms both individually and in any combination thereof.

Claims (45)

Formula (I):

[Where:
A is: i) an optionally substituted 5- or 6-membered cycloalkyl;
ii) an optionally substituted 5- or 6-membered heterocycloalkyl;
iii) optionally substituted 6-aryl; or iv) optionally substituted 5- or 6-membered heteroaryl;
T is a hydrogen atom or bonded to B i) forming an optionally substituted heterocycloalkyl; or ii) forming an optionally substituted heteroaryl;
B is an optionally substituted alkyl, an optionally substituted alkenyl, an optionally substituted alkynyl, an optionally substituted heteroalkyl, optionally substituted Optionally substituted cycloalkyl, optionally substituted heterocycloalkyl, optionally substituted alkylcycloalkyl, optionally substituted heteroalkylcycloalkyl, if An aryl which can be optionally substituted, an optionally substituted heteroaryl, an optionally substituted aralkyl, an optionally substituted heteroaralkyl or

And
W is N, alkyl, heteroalkyl, alkenyl, cycloalkyl, alkylcycloalkyl, heteroalkylcycloalkyl, aryl, heteroaryl, aralkyl or heteroaralkyl;
R ⁴ , if present, is a hydrogen atom, an optionally substituted alkyl, an optionally substituted heteroalkyl, an optionally substituted alkylcycloalkyl, optionally A heteroalkylcycloalkyl that may be substituted, an aryl that may be optionally substituted, an aralkyl that may be optionally substituted, a heteroaralkyl that may be optionally substituted, or Bonded to R ⁵ and together with W
v) an optionally substituted cycloalkyl;
vi) an optionally substituted heterocycloalkyl;
vii) optionally substituted aryl; or viii) forming optionally substituted heteroaryl;
R ⁵ , if present, is a hydrogen atom, an optionally substituted alkyl, an optionally substituted heteroalkyl, an optionally substituted heteroalkylcycloalkyl, An aryl that can be substituted by, an aralkyl that can be optionally substituted, a heteroaralkyl that can be optionally substituted, or bonded to R ⁴ and taken together with W;
v) an optionally substituted cycloalkyl;
vi) an optionally substituted heterocycloalkyl;
vii) optionally substituted aryl; or viii) forming optionally substituted heteroaryl;
E, if present, is heteroalkyl or heteroaralkyl;
G, if present, is alkyl, heteroalkyl, cycloalkyl, heterocycloalkyl, alkylcycloalkyl, heterocycloalkylalkyl, heteroaryl or heteroaralkyl;
K is

And
Here, R ¹ and R ² are each independently selected from C ₂ -C ₆ alkyl, C ₂ -C ₆ alkenyl, C ₂ -C ₆ alkynyl, C ₂ -C ₆ heteroalkyl, or together with N To form an optionally substituted heterocycloalkyl, an optionally substituted heteroaryl or an optionally substituted heteroaralkyl; and R ³ is a hydrogen atom. , C ₁ -or C ₂ alkyl, cyano or halogen atom]
Or a pharmacologically acceptable salt, solvate or hydrate thereof. A is

[Wherein X ¹ , X ² and X ³ are each independently selected from N, O, S, NR ^x , CR ^x or CR ^x R ^{x ′} , wherein R ^x and R ^{x ′} are each independently Hydrogen atom, halogen atom, = O, hydroxy, cyano, amino, nitro, mercapto, alkyl, alkenyl, alkynyl, heteroalkyl, cycloalkyl, heterocycloalkyl, alkylcycloalkyl, heteroalkylcycloalkyl, aryl, heteroaryl, Selected from Aralkyl or Heteroaralkyl]
A compound according to claim 1 wherein A is

[Where:
R ^x , R ^x1 , R ^x2 , R ^x3 , R ^{x ′} , R ^{x1 ′} , R ^{x2 ′} and R ^{x3 ′} are each independently a hydrogen atom, a halogen atom, ═O, hydroxy, cyano, amino, nitro, mercapto , Alkyl, alkenyl, alkynyl, heteroalkyl, cycloalkyl, heterocycloalkyl, alkylcycloalkyl, heteroalkylcycloalkyl, aryl, heteroaryl, aralkyl or heteroaralkyl; and X is NH, O or S]
A compound according to claim 1 or claim 2 selected from. A is

[Wherein Y ¹ , Y ² , Y ³ and Y ⁴ are each independently selected from N, O, S, NR ^y , CR ^y or CR ^y R ^{y ′} , where R ^y and R ^{y ′.} Each independently represents a hydrogen atom, a halogen atom, ═O, hydroxy, cyano, amino, nitro, mercapto, alkyl, alkenyl, alkynyl, heteroalkyl, cycloalkyl, heterocycloalkyl, alkylcycloalkyl, heteroalkylcycloalkyl, Selected from aryl, heteroaryl, aralkyl or heteroaralkyl]
A compound according to claim 1 wherein A is

[Where:
R ^y , R ^y1 , R ^y2 , R ^y3 , R ^y4 , R ^{y ′} , R ^{y1 ′} , R ^{y2 ′} , R ^{y3 ′} and R ^{y4 ′} are each independently a hydrogen atom, a halogen atom, ═O, hydroxy, Cyano, amino, nitro, mercapto, alkyl, alkenyl, alkynyl, heteroalkyl, cycloalkyl, heterocycloalkyl, alkylcycloalkyl, heteroalkylcycloalkyl, aryl, heteroaryl, aralkyl or heteroaralkyl]
5. A compound according to claim 1 or claim 4 selected from. A is

[Where:
R ^y1 , R ^y2 , R ^y3 and R ^y4 are each independently a hydrogen atom, a halogen atom, ═O, hydroxy, cyano, amino, nitro, mercapto, alkyl, alkenyl, alkynyl, heteroalkyl, cycloalkyl, heterocycloalkyl , Alkylcycloalkyl, heteroalkylcycloalkyl, aryl, heteroaryl, aralkyl or heteroaralkyl]
A compound according to any one of claims 1, 4 or 5. The compound according to claim 6, wherein R ^y1 is a hydrogen atom. R ^y2 is a hydrogen atom, a halogen atom or a _C 1 -C ₆ alkyl; and a compound according to claim 6 or claim 7 ^{R y4} is a hydrogen atom or a halogen atom. The compound according to any one of claims 6 to 8, wherein ^Ry2 is a hydrogen atom. The compound according to any one of claims 6 to 9, wherein ^Ry3 is a hydrogen atom or a halogen atom. The compound according to any one of claims 6 to 10, wherein ^Ry4 is a hydrogen atom or a halogen atom. The compound according to any one of claims 1 to 11, wherein T is a hydrogen atom. K is

A compound according to any one of claims 1 to 12 selected from. K is

14. A compound according to any one of claims 1 to 13 selected from. K is

15. A compound according to any one of claims 1-14. K is

15. A compound according to any one of claims 1-14. B is heteroalkyl, heteroaryl or

And
W is N, alkyl, alkenyl, cycloalkyl, alkylcycloalkyl, heteroalkylcycloalkyl, aryl, heteroaryl, aralkyl or heteroaralkyl;
R ⁴ , if present, is a hydrogen atom, alkyl, heteroalkyl, alkylcycloalkyl, heteroalkylcycloalkyl, aryl, aralkyl, heteroaralkyl, or bonded to R ⁵ and together with N
v) an optionally substituted cycloalkyl;
vi) an optionally substituted heterocycloalkyl;
vii) optionally substituted aryl; or viii) forming optionally substituted heteroaryl; and R ⁵ , if present, is a hydrogen atom, optionally substituted An optionally substituted heteroalkyl, an optionally substituted heteroalkyl, an optionally substituted heteroalkylcycloalkyl, an optionally substituted aryl, optionally substituted. An aralkyl which can be optionally substituted heteroaralkyl, or bonded to R ⁴ and taken together with N;
v) an optionally substituted cycloalkyl;
vi) an optionally substituted heterocycloalkyl;
17. A compound according to any one of claims 1 to 16 forming a vii) optionally substituted aryl; or viii) an optionally substituted heteroaryl. B is —Y ^aB —CO—L ^B — or —Y ^aB —CO—R ^cB —, where Y ^aB is a bond, C ₁ -C ₆ alkylene, C ₂ -C ₆ alkenylene or C ₂ -C _6. Alkynylene;
L ^B is a cycloalkyl, heterocycloalkyl, alkylcycloalkyl, heteroaryl alkyl cycloalkyl, optionally aryl which may be substituted, optionally heteroaryl may be substituted, aralkyl or heteroaralkyl; and R ^cB is an optionally substituted C ₁ -C ₈ alkyl, an optionally substituted C ₂ -C ₈ alkenyl or an optionally substituted C ₂ -C ₈ 18. A compound according to any one of claims 1 to 17 which is alkynyl. B is

19. A compound according to any one of claims 1-18. B is

And
Wherein R ⁴ is methyl, ethyl or isopropyl. The compound according to claim 1. B is

The compound according to any one of claims 1 to 17. E is ^{-Y aE -CO-L E -,} - Y aE -CO-R cE -, - Y aE -NR cE -CO-R dE -, - Y aE -NR cE -CO-L E -, - Y ^{aE -NR cE -CO-NR dE -L} E - or ^{-Y aE -NR cE -CS-NR dE} -L E - a, where in ^{Y aE} the _bond, C 1 -C _{6 alkylene,} C 2 -C It is ₆ alkenylene or _C 2 -C ₆ alkynylene;
R ^cE is a hydrogen atom, an optionally substituted C ₁ -C ₈ alkyl, an optionally substituted C ₂ -C ₈ alkenyl or an optionally substituted C _2- C ₈ alkynyl, provided that in —Y ^aE —CO—R ^cE —, R ^cE is not a hydrogen atom;
R ^dE is a hydrogen atom, an optionally substituted C ₁ -C ₈ alkyl, an optionally substituted C ₂ -C ₈ alkenyl or an optionally substituted C _2- be a C ₈ alkynyl; and L ^E is cycloalkyl, heterocycloalkyl, alkylcycloalkyl, heteroalkyl cycloalkyl, optionally aryl which may be substituted, optionally heteroaryl may be substituted, aralkyl Or a heteroaralkyl compound according to any one of claims 1 to 21. G is alkyl, ^{cycloalkyl, -Y aG -O-R cG,} -Y aG -CO-NR aG R bG, -Y aG -S-R cG, -Y aG -SO-R cG, -Y aG -SO _2- R ^cG , heteroaryl, alkylcycloalkyl or heterocycloalkyl, wherein Y ^aG is a bond, C ₁ -C ₆ alkylene, C ₂ -C ₆ alkenylene or C ₂ -C ₆ alkynylene;
R ^aG is a hydrogen atom, C ₁ -C ₆ alkyl, C ₂ -C ₆ alkenyl, C ₂ -C ₆ alkynyl, or bonded to R ^bG to form a 4- to 10-membered cycloalkyl or heterocycloalkyl. Forming;
R ^bG is a hydrogen atom, C ₁ -C ₆ alkyl, C ₂ -C ₆ alkenyl, C ₂ -C ₆ alkynyl, or bonded to R ^aG to form a ^4- to 10-membered cycloalkyl or heterocycloalkyl. And R ^cG is an optionally substituted C ₁ -C ₈ alkyl, an optionally substituted C ₂ -C ₈ alkenyl or an optionally substituted C ₂ the compound according to any one of claims 1 to 22 -C is ₈ alkynyl. The compound is of formula (II):

Indicated by
Where
Y ¹ is N, CH or CF;
K is

Chosen from;
R ³ is a hydrogen atom or methyl;
R ^y2 is a hydrogen atom, a halogen atom or C ₁ -C ₆ alkyl;
R ^y3 is a hydrogen atom, a halogen atom, C ₁ -C ₆ alkyl or C ₁ -C ₆ heteroalkyl;
R ^y4 is a hydrogen atom or a halogen atom;
The ^{B -Y aB -CO-L B -} , - Y aB -CO-R cB - or

And
Where Y ^aB is a bond;
R ^cB is an optionally substituted C ₁ -C ₈ alkyl or an optionally substituted C ₂ -C ₈ alkenyl;
L ^B is cycloalkyl, heterocycloalkyl, optionally aryl which may be substituted, optionally heteroaryl or heteroaralkyl may be substituted;
W is alkyl or N;
R ⁴ , if present, is a hydrogen atom, alkyl, alkylcycloalkyl, heteroalkylcycloalkyl, aralkyl or heteroaralkyl;
R ⁵ , if present, is a hydrogen atom or alkyl;
E is ^{-Y bE -NR dE -CO-L E} - a, where in ^{Y bE} is _bound, be a C 1 -C ₆ alkylene or _C 2 -C ₆ alkenylene;
R ^dE represents a hydrogen _atom, a C 1 -C ₈ alkyl or _C 2 -C ₆ alkenyl; are substituted and ^{L E} is cycloalkyl, heterocycloalkyl, aryl which may be optionally substituted, optionally 23. A compound according to any one of claims 1, 4-6, 12, 17 or 22 which is heteroaryl or heteroaralkyl. The compound is of formula (III) or (IV):

Indicated by
Where
Y ¹ is N, CH or CF;
K is

Chosen from;
R ³ is a hydrogen atom or methyl;
R ^y2 is a hydrogen atom, a halogen atom or C ₁ -C ₆ alkyl;
R ^y3 is a hydrogen atom, a halogen atom, C ₁ -C ₆ alkyl or C ₁ -C ₆ heteroalkyl;
R ^y4 is a hydrogen atom or a halogen atom, C ₁ -C ₆ alkyl or C ₁ -C ₆ heteroalkyl;
E is ^{-Y aE -NR cE -CO-R dE} -, - Y aE -NR cE -CO-L E -, - Y aE -NR cE -CO-NR dE -L E - or ^-Y aE ^{-NR cE -CS-NR} dE -L ^E - a, where in ^{Y aE} is a bond or _C 1 -C ₆ alkylene;
R ^cE is a hydrogen atom, C ₁ -C ₆ alkyl or C ₂ -C ₆ alkenyl;
R ^dE is a hydrogen atom, an optionally substituted C ₁ -C ₈ alkyl, an optionally substituted C ₂ -C ₈ alkenyl or an optionally substituted C _2- A C ₈ alkynyl group;
L ^E is cycloalkyl, heterocycloalkyl, alkylcycloalkyl, heteroaryl alkyl cycloalkyl, optionally aryl which may be substituted, optionally heteroaryl may be substituted, aralkyl or heteroaralkyl;
G, if present, is cycloalkyl, —Y ^aG —CO—NR ^{aGR bG} , —Y ^aG —O—R ^eG , —Y ^aG —S—R ^eG , —Y ^aG —SO—R ^eG or — Y ^aG —SO ₂ —R ^eG where Y ^aG is a bond or C ₁ -C ₆ alkylene;
R ^aG is a hydrogen atom or C ₁ -C ₆ alkyl;
R ^bG is a hydrogen atom or C ₁ -C ₆ alkyl; and R ^eG is C ₁ -C ₆ alkyl, any one of claims 1, 4-6, 12, 13, 17-20, 22 or 23. A compound according to claim. The compound is of formula (V):

Indicated by
^{Wherein, Y 1, K, R 3} , R y2, R y3, R y4 and ^{L E} are compounds according to claim 25 are as defined in claim 25. The compound is of formula (VI):

Indicated by
^{Wherein, Y 1, K, R 3} , R y2, R y3, R y4 and ^{L E} are compounds according to claim 25 are as defined in claim 25. ^{28. A} compound according to any one of claims 24 to 27 wherein ^Ry3 is a hydrogen atom. 29. The compound according to any one of claims 24 to 28, wherein ^Ry2 is a hydrogen atom. The compound is of formula (VII):

Indicated by
Where
Y ¹ is N, CH or CF;
K is

Chosen from;
R ^y4 is a hydrogen atom, a halogen atom, C ₁ -C ₆ alkyl or C ₁ -C ₆ heteroalkyl;
E is ^-Y aE ^{-CO-L E} - a, where in ^{Y aE} is a bond or _C 1 -C ₆ alkylene;
L ^E is heterocycloalkyl; and G is alkyl cycloalkyl, heterocycloalkylalkyl, claim 1,4～6,9,10,12,13,17,21,22 or 23 aryl or heteroaryl Earl A compound according to any one of the above. The compound according to claim 30, wherein ^Ry4 is a hydrogen atom or a halogen atom. Stereogenic centers R ³ is attached is (R) is in the configuration; and the compound according to any one of claims 1 to 31 wherein R ³ is methyl. Compound, a standard in vitro BK B1 receptor - The compound according to any one of claims 1 to 32 showing a 500nM to or lower _{IC 50} than in mediating assay, a salt, solvate or hydrate.   34. A compound according to any one of claims 1 to 33, wherein the compound is selected from compounds 1 to 282 of Table 1.   35. A pharmaceutical composition comprising one or more compounds according to any one of claims 1 to 34 and optionally at least one carrier substance, excipient and / or auxiliary.   36. The pharmaceutical composition according to claim 35, wherein the pharmaceutical composition is prepared as an aerosol, cream, gel, pill, capsule, syrup, solution, transdermal therapeutic system, suppository or pharmaceutical device.   Use of a compound or pharmaceutical composition according to any one of claims 1 to 36 for the manufacture of a medicament for the treatment and / or prevention of a disease or condition.   38. Use according to claim 37, wherein the condition or disease is responsive to BK B1R modulation.   39. Use according to claim 37 or claim 38, wherein the condition or disease is selected from the group comprising inflammatory diseases, immune disorders and pain.   40. Use according to claim 39, wherein the inflammatory disease or immune disorder is selected from the group comprising inflammatory bowel disease, rheumatoid arthritis, gouty arthritis, atherosclerosis and related fibrotic conditions.   40. Use according to claim 39, wherein the pain is selected from the group comprising visceral pain, neuropathic pain, complex regional pain syndrome CRPS and inflammatory pain.   37. At least one species according to any one of claims 1-36, under conditions and in an amount sufficient to detectably detect the binding of DAKD, KD and DABK to the BK B1 receptor. A method of inhibiting the binding of DAKD, KD and DABK to the BK B1 receptor in vitro comprising contacting with a compound of claim 1 or a pharmaceutically acceptable salt, solvate or hydrate thereof. (A) a sample of tissue presumed to contain a BK B1 receptor is detectably labeled with a compound according to any one of claims 1 to 36 and allowing the compound to bind to the BK B1 receptor And (b) detecting a compound bound to the BK B1 receptor or detecting binding of the compound to the BK B1 receptor in a tissue, preferably a tissue in vitro. A method for locating or detecting a BK B1 receptor in a section.   44. The method according to claim 43, wherein the compound is radiolabeled, fluorescent-labeled or luminescent labeled, or labeled with an antibody.   37. A method for treating a patient in need of such treatment comprising administering a compound or pharmaceutical composition according to any one of claims 1-36.