UA103034C2 - Picolinamide derivatives as kinase inhibitors - Google Patents

Abstract

New picolinamide compounds, compositions and methods of inhibition of Provirus Integration of Maloney Kinase (PIM kinase) activity associated with tumorigenesis in a human or animal subject are provided. In certain embodiments, the compounds and compositions are effective to inhibit the activity of at least one PIM kinase. The new compounds and compositions may be used either alone or in combination with at least one additional agent for the treatment of a serine/threonine kinase-or receptor tyrosine kinase-mediated disorder, such as cancer.

Description

"

Asia Ripaptasoi bip 26(3):364-8 (2005); Etsi ei a. 2005; her ey aYi 2006). In addition, it is believed that the mutational activation of some well-known oncogenes in hematopoietic malignant neoplasms at least partially manifests its effects with the help of Rit kinases. For example, the targeted downregulation of Rit expression disrupts the vital activity of hematopoietic cells transformed with the help of RIZ and VSK/AVI. (Adat ei ai. 2006). Thus, inhibitors of Rit!, 2 and Z should be used for the treatment of these malignant neoplasms. In addition to potential applications in the treatment of cancer and myeloproliferative diseases, such an inhibitor should be useful in regulating the proliferation of immune cells in other pathological conditions, such as autoimmune diseases, allergic reactions, and organ transplant rejection syndromes. In favor of such a conclusion, the data show that the differentiation of TP1 helper T cells with the help of

I--12 ii IEM-a leads to the induction of expression of both Riti and 2 (Ano Tei ai., "Ehrgezviop oi pitap Rit

Tatiiu depes with zeIesiimeIzu ir-geashaiea Bu suioKipe5 rgootoiypa T Peireg iure 1, ryi poi T Peireg iure 2, sei ayegepiayop, " Ittypoiodu 116: 82-88 (2005)). In addition, the expression of Rit kinases is repressed in both cell types by immunosuppressive TOEs - (Ayo et al. 2005) These results show that Rit kinases are involved in the early process of differentiation of helper T cells, which coordinates immune responses in autoimmune diseases, allergic reactions and rejection of tissue transplants.

There remains a need for compounds that suppress capillary proliferation, suppress tumor growth, treat cancer, modulate cell cycle arrest, and/or inhibit molecules such as Rit!, Rit2,

Rit3, and in pharmaceutical preparations and medicines that contain such a compound. Also needed are methods of administering such compounds, pharmaceuticals, and drugs to patients or subjects in need.

Brief summary of the essence of the invention

This invention relates to new compounds of formula I and to their stereoisomers, tautomers and pharmaceutically acceptable salts

H5 in Hn r i i ai y: ? what

Xz:x rah (c) Vu

Ho in which

X1, X2, X3Z and X4 are independently selected from the group consisting of SKZ and M; provided that at least one, but not more than two of X1, X2, X3Z and X4 denote M;

In selected from the group including cycloalkyl, partially unsaturated cycloalkyl and heterocycloalkyl, where each representative of the specified group can contain up to 4 substituents; 22 and 73 are independently selected from the group consisting of SKT12 and M; provided that no more than one of 22 and 273 may denote M;

CI is selected from the group consisting of hydrogen, MNEC, halogen, hydroxy, alkyl, cyano, and nitro;

K2 and Ky: in each case independently selected from the group consisting of hydrogen, halogen, hydroxy, nitro, cyano, ZOZN and substituted or unsubstituted alkyl, alkenyl, alkynyl, alkoxy, amino, cycloalkyl, heterocycloalkyl and partially saturated cycloalkyl;

EZ is selected from the group consisting of hydrogen, SOK" and substituted or unsubstituted alkyl, cycloalkyl, heterocyclyl, aryl and heteroaryl;

Va is selected from the group consisting of alkyl, substituted alkyl, alkyl, substituted alkyl, amino, substituted amino, and alkylamino; and

AB5 represents a group selected from the group consisting of substituted or unsubstituted aryl, C3-C7-cycloalkyl, heteroaryl, partially unsaturated cycloalkyl, and alkyl, in which each said substituted group C5 may contain up to 4 substituents selected from the group consisting of halogen, cyano group, amino group,

C1-C6-alkyl, C3-C6-cycloalkyl, alkoxy group, nitro group, carboxy group, carbonyl, carbolic group, aminocarboxy group, substituted aminocarbonyl, aminosulfonyl, substituted aminosulfonyl and alkoxyalkyl.

Some embodiments relate to new compounds of formula I or their stereoisomer, tautomer or pharmaceutically acceptable salt, in which X2 denotes M and Xy, Xz and Xy. denote CE".

Some embodiments relate to novel compounds of formula I or a stereoisomer, tautomer, or pharmaceutically acceptable salt thereof, wherein B2 is selected from the group consisting of hydrogen, methyl, ethyl,

halogen, cyano group.

Some variants of implementation refer to compounds of formula I or their stereoisomer, tautomer or pharmaceutically acceptable salt, in which 2" and 7z denote SKi".

Some embodiments relate to compounds of formula I or a stereoisomer, tautomer, or pharmaceutically acceptable salt thereof, wherein K:i2 is selected from the group consisting of hydrogen, halogen, methyl, ethyl, and cyano.

Other embodiments relate to new compounds and their stereoisomers, tautomers and pharmaceutically acceptable salts of formula II

Av

Vu2 in Nn M and ! dSh M Zh

Wu2

In o); IN,

I am in which

In selected from the group consisting of cyclohexyl, partially unsaturated cyclohexyl and heterocyclo-

St-alkyl, where each representative of the specified group can contain up to 4 substituents;

K1 is selected from the group consisting of hydrogen, MNEZ, halogen, hydroxy group, alkyl, C3-C4-cycloalkyl, cyano group and nitro group;

Wig in each case is independently selected from the group consisting of hydrogen, halogen, hydroxy, amino, nitro, cyano, 3ZZN, and substituted or unsubstituted alkyl, alkenyl, alkynyl, alkoxy, amino, cycloalkyl, heterocycloalkyl, and partially saturated cycloalkyl;

EZ is selected from the group consisting of hydrogen, SOK" and substituted or unsubstituted alkyl, cycloalkyl, heterocyclyl, aryl and heteroaryl;

Va is selected from the group consisting of alkyl, substituted alkyl, alkyl, substituted alkyl, amino, substituted amino, and alkylamino; and

B5 represents a group selected from the group consisting of hydrogen and substituted or unsubstituted alkyl, C1-cycloalkyl, aryl and heteroaryl, in which each said substituted group Co may contain up to 4 substituents selected from the group consisting of halogen, cyano, amino, C--Ca-alkyl, C3-Cv-cycloalkyl, alkoxy group, nitro group, carboxy group, carbonyl, carb carboxy group, aminocarboxy group, substituted aminocarbonyl, aminosulfonyl, substituted aminosulfonyl and alkoxyalkyl.

Some embodiments refer to the compounds of formulas I or II or their stereoisomer, tautomer or pharmaceutically acceptable salt, in which M is selected from the group consisting of substituted or unsubstituted cycloalkyl, cycloalkenyl, piperidinyl and piperazinyl, where each representative of said group contains up to 4 substituents. In some embodiments, Y contains up to 4 substituents selected from the group consisting of cyano, nitro, halogen, hydroxy, amino, alkoxy, substituted amino, C1-C4-alkyl, C1-C4-haloalkyl, and C3-C4-cycloalkyl . In other embodiments, U contains up to 4 substituents selected from the group consisting of methyl, propyl, isopropyl, ethyl, hydroxy, amino, halogen, monohalo-Ci-C3-alkyl, trihalo-C:-C3-alkyl, and dihalo-C :1-C3-alkyl.

Some embodiments relate to novel compounds of formula II or a stereoisomer, tautomer, or pharmaceutically acceptable salt thereof, wherein M is selected from the group consisting of substituted or unsubstituted cyclohexyl, cyclohexynyl, and piperidinyl, wherein each member of said group contains up to 4 substituents.

Some embodiments relate to novel compounds of formula II or a stereoisomer, tautomer, or pharmaceutically acceptable salt thereof wherein U contains up to 4 substituents independently selected from the group consisting of hydrogen, cyano, nitro, halogen, hydroxy, amino, alkoxy, substituted amino , C-C-alkyl, C-C-haloalkyl and C-C-cycloalkyl. In some embodiments, the substituents are independently selected from the group consisting of methyl, propyl, isopropyl, ethyl, hydroxy, amino, halogen, monohalo-C1-C3-alkyl, trihalo-C1-C3-alkyl, and dihalo-C1-C3-alkyl.

Some embodiments relate to compounds of formula III or a stereoisomer, tautomer, or pharmaceutically acceptable salt thereof, wherein K:i2 is selected from the group consisting of hydrogen, halogen, methyl, ethyl, and cyano.

Some embodiments relate to compounds of formulas I or II or a stereoisomer, tautomer or pharmaceutically acceptable salt thereof, wherein M is selected from the group consisting of substituted or unsubstituted cyclohexyl, cyclohexenyl, piperidinyl, piperazinyl, wherein said group M may contain up to 3 substituents , selected from the group including methyl, ethyl, hydroxy group, amino group and methoxy group; CI is selected from the group consisting of hydrogen and an amino group; and K1I2 independently in each case represents hydrogen, halogen, or methyl.

Some embodiments relate to compounds of formula II or a stereoisomer, tautomer, or pharmaceutically acceptable salt thereof, wherein M is selected from the group consisting of substituted cyclohexyl, cyclohexenyl, piperidinyl, and piperazinyl; K1 is selected from the group consisting of hydrogen, -MNAI, halogen, Si1-

C4-alkyl, C3-Cb4-cycloalkyl and CM; K12 in each case is independently selected from the group consisting of hydrogen, halogen, C-C-alkyl and amino; and K5 is selected from the group consisting of substituted or unsubstituted phenyl, cyclohexyl, cyclopentyl, thiazole, pyridyl, pyrimidyl and pyrazinyl, wherein the group K5 may contain up to 3 substituents selected from the group consisting of halogen, hydrogen, methyl, substituted aminocarbonyl and Alkoxy group.

In a typical embodiment, a compound of formulas I or II or a stereoisomer, tautomer or pharmaceutically acceptable salt thereof is selected from the group consisting of M-(4-((18, ZA, 48, 55)-3-amino-4-hydroxy-

B-methylcyclohexyl)pyridin-3-yl)-6-(2,6-difluorophenyl)-5-fluoropicolinamide, M-(4-(1B8, 35, 55)-3-amino-5-methylcyclohexyl)pyridin-3- yl)-6-(2,6-difluorophenyl)-5-fluoropicolinamide. M-(4-(Z3А8, 48, 55)-3-amino-4-hydroxy-5-methylpiperidin-1-yl)pyridin-3-yl)-6-(2,6-difluorophenyl)-5-fluoropicolinamide, 3-amino-M-(4-(ZA, AB, 55)-3-amino-4-hydroxy-5-methylpiperidin-1-yl)pyridin-3-yl)-6-(2,6-difluorophenyl)- 5-fluoropicolinamide, and M-(4-(18, 35)-3-aminocyclohexyl)pyridin-3-yl)-6-(2,6-difluorophenyl)-5-fluoropicolinamide, and 3-amino-M-(4 -(18,35)-3-aminocyclohexyl)pyridin-3-yl)-6-(2,6-difluorophenyl)-5-fluoropicolinamide.

In other objects, the present invention relates to methods of treating disorders associated with proviral integration of Meloni kinase (RIM kinase) in a human or animal in need of such treatment, comprising administering to said subject a compound of formula I! or ITS in an amount effective to inhibit RIM activity in the subject.

In other objects, the present invention relates to methods of treating disorders associated with RIM in a human or animal in need of such treatment, comprising administering to said subject a compound of formula I or II in an amount effective to reduce or prevent growth tumors in the subject.

In other objects, the present invention relates to methods of treating disorders associated with RIM in a human or animal in need of such treatment, comprising administering to said subject a compound of formula I or II in an amount effective to reduce or prevent growth tumor in the subject, in combination with at least one additional agent for the treatment of cancer.

In other objects, the present invention relates to a therapeutic composition containing at least one compound of formula I or II in combination with one or more additional agents intended for the treatment of cancer, which are commonly used for the treatment of cancer.

The compounds of this invention can be used to treat cancers, including hematopoietic malignancies, carcinomas (e.g., lung, liver, pancreas, ovary, thyroid, bladder, or colon), melanomas, myeloid disorders (e.g., myeloid leukemia, multiple myeloma and erythroleukosis), adenomas (for example, villous adenoma of the colon), sarcomas (for example, osteosarcoma), autoimmune diseases, allergic reactions and organ transplant rejection syndromes.

This invention also relates to the compositions, methods of application and methods of preparation described in the detailed description of the invention.

Description of drawings

The above objects and many other advantages of the present invention will be more easily appreciated when they are better understood from the following detailed description together with the accompanying drawings, in which the following are shown:

In fig. 1 presents the dependence for the effectiveness of the compound of example 99 according to the data of the KM511-Ins xerograft model described in example 144.

In fig. 2 shows the dependence for the effectiveness of the compound of example 70 according to the data of the KM511-Ins xerograft model described in example 144.

In fig. C shows the dependence for the effectiveness of the compound of example 96 according to the data of the KM511-Ins xerograft model described in example 144.

Detailed description of the invention

Another object of the present invention are new compounds and their stereoisomers, tautomers and pharmaceutically acceptable salts of formula I:

AB in NM

M third

Khzh ri Vi

Ho in which

X1, X2, X3Z and X4 are independently selected from the group consisting of SKZ and M; provided that at least one, but not more than two of X1, X2, X3Z and X4 denote M;

In selected from the group including cycloalkyl, partially unsaturated cycloalkyl and heterocycloalkyl, where each representative of the specified group can contain up to 4 substituents; 22 and 73 are independently selected from the group consisting of SKT12 and M; provided that no more than one of 22 and 273 may denote M;

KI is selected from the group consisting of hydrogen, MNH, halogen, hydroxy, alkyl, cyano, and nitro;

K2 and Ky: in each case independently selected from the group consisting of hydrogen, halogen, hydroxy, nitro, cyano, ZOZN and substituted or unsubstituted alkyl, alkenyl, alkynyl, alkoxy, amino, cycloalkyl, heterocycloalkyl and partially saturated cycloalkyl;

EZ is selected from the group consisting of hydrogen, SOK" and substituted or unsubstituted alkyl, cycloalkyl, heterocyclyl, aryl and heteroaryl;

Ba is selected from the group consisting of alkyl, substituted alkyl, alkoxy, substituted alkoxy, amino, substituted amino, and alkylamino; and

AB5 represents a group selected from the group consisting of substituted or unsubstituted aryl, C3-C7-cycloalkyl, heteroaryl, partially unsaturated cycloalkyl, and alkyl, in which each said substituted group C5 may contain up to 4 substituents selected from the group consisting of halogen, cyano group, amino group,

C1-C6-alkyl, C3-C6-cycloalkyl, alkoxy group, nitro group, carboxy group, carbonyl, carbolic group, aminocarboxy group, substituted aminocarbonyl, aminosulfonyl, substituted aminosulfonyl and alkoxyalkyl.

Some variants of implementation refer to new compounds of formula I or their stereoisomer, tautomer or pharmaceutically acceptable salt, in which X2 denotes M and Xy, Xz and X denote SK2.

Some embodiments relate to new compounds of formula I or their stereoisomer, tautomer or pharmaceutically acceptable salt, in which K2 is selected from the group consisting of hydrogen, methyl, ethyl, halogen, cyano group.

Some variants of implementation relate to compounds of formula I or their stereoisomer, tautomer or pharmaceutically acceptable salt, in which 2" and 7z denote SK12.

Some embodiments relate to compounds of formula I or a stereoisomer, tautomer, or pharmaceutically acceptable salt thereof, wherein K:i2 is selected from the group consisting of hydrogen, halogen, methyl, ethyl, and cyano.

Other embodiments relate to new compounds and their stereoisomers, tautomers, and pharmaceutically acceptable salts of formula I

IN

Ag in N M

AND

M. p

OO | Vi" with V.

I am in which

In selected from the group consisting of cyclohexyl, partially unsaturated cyclohexyl and heterocyclo-

St-alkyl, where each representative of the specified group can contain up to 4 substituents;

K1 is selected from the group consisting of hydrogen, MNEZ, halogen, hydroxy group, alkyl, C3-C4-cycloalkyl, cyano group and nitro group;

Wig is in each case independently selected from the group consisting of hydrogen, halogen, hydroxy, amino, nitro, cyano, ZOZZN, and substituted or unsubstituted alkyl, alkenyl, alkynyl,

alkoxy group, amino group, cycloalkyl, heterocycloalkyl and partially saturated cycloalkyl;

EZ is selected from the group consisting of hydrogen, SOK" and substituted or unsubstituted alkyl, cycloalkyl, heterocyclyl, aryl and heteroaryl;

Va is selected from the group consisting of alkyl, substituted alkyl, alkyl, substituted alkyl, amino, substituted amino, and alkylamino; and

B5 represents a group selected from the group consisting of hydrogen and substituted or unsubstituted alkyl, C1-cycloalkyl, aryl and heteroaryl.

Some embodiments refer to the compounds of formulas I or II or their stereoisomer, tautomer or pharmaceutically acceptable salt, in which M is selected from the group consisting of substituted or unsubstituted cycloalkyl, cycloalkenyl, piperidinyl and piperazinyl, where each representative of said group contains up to 4 substituents. Some embodiments relate to compounds of formulas I or II, or a stereoisomer, tautomer, or pharmaceutically acceptable salt thereof, wherein X is selected from the group consisting of substituted or unsubstituted cyclohexyl, cyclohexynyl, and piperidinyl, wherein each member of said group contains up to 4 substituents. In some embodiments, Y contains up to 4 substituents selected from the group consisting of hydrogen, cyano, nitro, halogen, hydroxy, amino, alkoxy, substituted amino, C--C a-alkyl, C--C--haloalkyl, and C 3-C --cycloalkyl. In other embodiments, U contains up to 4 substituents selected from the group consisting of methyl, propyl, isopropyl, ethyl, hydroxy, amino, halogen, monohalo-C1-C3-alkyl, trihalo-C1-C3-alkyl, and dihalo-

C.-C3-alkyl.

Some variants of implementation refer to compounds of formulas | or II or their stereoisomer, tautomer or pharmaceutically acceptable salt, in which EC: denotes hydrogen, an amino group or fluorine. One embodiment relates to compounds of formula II selected from Table I or Table II.

Some embodiments relate to compounds of formulas I or II, or a stereoisomer, tautomer, or pharmaceutically acceptable salt thereof, wherein K5 is selected from the group consisting of substituted or unsubstituted aryl, C5-C6-cycloalkyl, heteroaryl, partially unsaturated C5-C6-cycloalkyl, and C1-C4 alkyl, where each specified group may contain up to 4 substituents selected from the group consisting of halogen, cyano, amino, C1-C6-alkyl, C3-C6-cycloalkyl, alkoxy, nitro, carboxy, carbonyl, carbocarboxy, aminocarboxy , substituted aminocarbonyl, aminosulfonyl, substituted aminosulfonyl and alkoxyalkyl. Some embodiments relate to compounds of formulas I or II or a stereoisomer, tautomer or pharmaceutically acceptable salt thereof, wherein K5 is substituted or unsubstituted phenyl, where the phenyl group may contain up to 4 substituents selected from the group consisting of hydrogen, cyano, nitro, halogen, hydroxy group, amino group, alkoxy group, substituted amino group, C1-C4-alkyl, C1-C4-haloalkyl and C3-C4-cycloalkyl. Some embodiments relate to compounds of formulas I or II or their stereoisomer, tautomer or pharmaceutically acceptable salt in which

AB5 denotes 2,6-difluorophenyl.

Some embodiments relate to compounds of formulas I or II or their stereoisomer, tautomer, or pharmaceutically acceptable salt, wherein K:2 is selected from the group consisting of hydrogen, halogen, methyl, ethyl, and cyano. Some embodiments refer to compounds of formulas I or II or their stereoisomer, tautomer or pharmaceutically acceptable salt, in which X is selected from the group consisting of substituted or unsubstituted cyclohexyl, cyclohexenyl, piperidinyl, piperazinyl, in which said group M can contain up to 3 substituents , selected from the group including methyl, ethyl, hydroxy group, amino group and methoxy group; K1 is selected from the group consisting of hydrogen and an amino group; and K1I2 independently in each case represents hydrogen, halogen, or methyl.

Some embodiments relate to compounds of formula II or a stereoisomer, tautomer, or pharmaceutically acceptable salt thereof, wherein M is selected from the group consisting of substituted cyclohexyl, cyclohexenyl, piperidinyl, and piperazinyl; K1 is selected from the group consisting of hydrogen, -MNa, halogen, Si

C1-Ca-alkyl, C3-b4-cycloalkyl and CM; K12 in each case is independently selected from the group consisting of hydrogen, halogen, C-C-alkyl and amino; and K5 is selected from the group consisting of substituted or unsubstituted phenyl, cyclohexyl, cyclopentyl, thiazole, pyridyl, pyrimidyl and pyrazinyl, wherein the group K5 may contain up to 3 substituents selected from the group consisting of halogen, hydrogen, methyl, substituted aminocarbonyl and Alkoxy group.

The best embodiment of the present invention is a compound of the formula (I), in which M denotes cyclohexyl, containing up to 3 substituents, these substituents are preferably selected from the group including a hydroxy group, an amino group, C1-C4-alkyl or C1-C4-haloalkyl and more preferably selected from the group consisting of methyl, hydroxy, amino, and C3, and most preferably selected from the group consisting of methyl, amino, and hydroxy; K: denotes hydrogen, MH" or halogen (better, K: denotes hydrogen, an amino group or fluorine, even better Ki denotes hydrogen); Kig2 all independently represent hydrogen or halogen (it is better if each Ki represents hydrogen, chlorine or fluorine); Co represents cyclohexyl, phenyl, or pyridyl, wherein said cyclohexyl, said phenyl, and said pyridyl all independently contain up to C substituents selected from the group consisting of halogen, hydroxy, C1-C6-alkyl, and C1-C6-alkyl (preferably, if K5 represents pyridyl or phenyl, each of which independently contains up to C substituents selected from the group consisting of halogen, hydroxy, C1-C4-alkyl or C1-C4-alkyl, more preferably,

if K5 represents phenyl containing up to C substituents selected from the group consisting of halogen, hydroxy, C1-C4-alkyl and C1-C4-alkyl, preferably phenyl containing up to C substituents selected from the group consisting of fluorine, hydroxy group, methyl, ethyl, methoxy group, or propoxy group, preferably if K5 is 2,6-difluorophenyl.

In another preferred embodiment, the present invention relates to a compound of formula II, in which Y represents piperidinyl substituted with methyl, hydroxy group and amino group; WHO! denotes hydrogen, MH", or fluorine; Cig is in each case independently selected from the group consisting of hydrogen and halogen; and K5 represents pyridyl, fluoropyridyl, cyclohexyl, or phenyl, where said phenyl contains up to 3 substituents selected from the group consisting of fluorine, hydroxy, and methyl, preferably if K5 represents difluorophenyl. In another preferred embodiment, Y preferably denotes C3-amino-4-hydroxy-5-methylpiperidin-1-yl; Ki represents hydrogen and Co represents 2,6-difluorophenyl.

In a typical embodiment, preferred compounds of formulas I or II or a stereoisomer, tautomer or pharmaceutically acceptable salt thereof are selected from the group consisting of M-(4-(35,55)-3-amino-5-methylcyclohexyl)pyridin-3-yl) -6-(2,6-difluorophenyl)-5-fluoropicolinamide; Z-amino-M-(4-(18, ZB, 45, 55)-

3-amino-4-hydroxy-5-methylcyclohexyl)pyridin-3-yl)-6-(2,6-difluorophenyl)picolinamide; M-(4-(3A, 48, 55)-3-amino-4-hydroxy-5-methylpiperidin-1-yl)pyridin-3-yl)-6-(2,6-difluorophenyl)-5-fluoropicolinamide; M-(4-(ZB, AB, 55)-3-amino-4-hydroxy-5-methylpiperidin-1-yl)pyridin-3-yl)-6-(2,6-difluorophenyl)-5-fluoropicolinamide; 3-amino-M-(4-(3А8, 48, 55)-3-amino-4-hydroxy-5-methylpiperidin-1-yl)pyridin-3-yl)-6-(2,6-difluorophenyl)- 5-fluoropicolinamide; M-(4-(3А8, 48, 55)-3-amino-4-hydroxy-5-methylpiperidin-1-yl)pyridin-3-yl)-6-(2,6-difluoro-3-methylphenyl)- 5-fluoropicolinamide; 3-amino-N-(4-(18,35)-3-aminocyclohexyl)pyridin-3-yl)-6-(2,6-difluorophenyl)-5-fluoropicolinamide; M-(4-((35)-3-aminocyclohexyl)pyridin-3-yl)-6-(2,6-difluorophenyl)-5-fluoropicolinamide; M-(4-(18, ZA, 48, 55)-3-amino-4-hydroxy-5--methylcyclohexyl)pyridin-3-yl)-6-(2,6-difluorophenyl)-5-fluoropicolinamide; M-(4-(18, 35, 55)-3-amino-5-methylcyclohexyl)pyridin-3-yl)-6-(2,6-difluorophenyl)-5-fluoropicolinamide); M-(4-(Z38, 48, 55)-3-amino-4-hydroxy-5-methylpiperidin-1-yl)pyridin-3-yl)-6-(2,6-difluorophenyl)-5-fluoropicolinamide; M-(4-18, /35)-3-aminocyclohexyl)pyridin-3-yl)-6-(2,6-difluorophenyl)-5-fluoropicolinamide; 3-amino-M-(4-(ZA, AB, 55)-3-amino-4-hydroxy-5-methylpiperidin-1-yl)pyridin-3-yl)-6-(2,6-difluorophenyl)- 5- fluoropicolinamide; and 3-amino-N-(4-(18,35)-3-aminocyclohexyl)pyridin-3-yl)-6-(2,6-difluorophenyl)-5-fluoropicolinamide.

In other objects, the present invention relates to methods of treating disorders associated with the proviral integration of Meloni kinase (RIM kinase) in a human or animal in need of such treatment, comprising administering to said subject a compound of formula I or I in an amount , effective for inhibiting the activity of RIM in the subject. In a preferred embodiment, the present invention relates to a method of treating a pathological condition by modulating the activity of Maloni's proviral integration kinase (RIM kinase), which includes administering to a patient in need of such treatment a compound of formula I in an effective amount.

In other objects, the present invention relates to methods of treating disorders associated with RIM in a human or animal in need of such treatment, comprising administering to said subject a compound of formula I or II in an amount effective to reduce or prevent growth tumors in the subject. In other objects, this invention relates to methods of treatment of disorders associated with

RIM, in a human or animal in need of such treatment, comprising administering to said subject a compound of formula I or II in an amount effective to reduce or prevent tumor growth in the subject, in combination with at least one additional agent for the treatment cancer

In another object, the present invention relates to a therapeutic composition containing at least one compound of formula I or II in combination with one or more additional agents intended for the treatment of cancer, which are commonly used for the treatment of cancer. Thus, the present invention relates to a pharmaceutical composition containing a compound of formula I or formula

I. The best embodiment of this object is a pharmaceutical composition containing a compound selected from the group consisting of M-(4-((35, 55)-3-amino-5-methylcyclohexyl)pyridin-3-yl)- 6-(2,6-difluorophenyl)-5-fluoropicolinamide; 3-amino-M-(4-(18, ZA, 45, 55)-3-amino-4-hydroxy-5-methylcyclohexyl)pyridin-3-yl)-6-(2,6-difluorophenyl)picolinamide; M-(4-(3А8, 48, 55)-3-amino-4-hydroxy-5-methylpiperidin-1-yl)pyridin-3-yl)-6-(2,6-difluorophenyl)-5-fluoropicolinamide; 3-amino-M-(4-(18, 35)-33-aminocyclohexyl)pyridin-3-yl)-6-(2,6-difluorophenyl)-5-fluoropicolinamide; M-(4-((35)-3-aminocyclohexyl)pyridin-3-yl)-6-(2,6-difluorophenyl)-5-fluoropicolinamide; M-(4-(18, ZB, AB, 55)-3-amino-4-hydroxy-5-methylcyclohexyl)pyridin-3-yl)-6-(2,6-difluorophenyl)-5-fluoropicolinamide; M-(4-(18, 35, 55)-3-amino-5-methylcyclohexyl)pyridin-3-yl)-6-(2,6-difluorophenyl)-5-fluoropicolinamide); M-(4-(ЗВ, 48, 55)-3-amino-4-hydroxy-o-methylpiperidin-1-yl)/pyridin-3-yl)-6-(2,6-difluorophenyl)-5-fluoropicolinamide ; M-(4-(1А8, 35)-3-aminocyclohexyl)pyridin-3-yl)-6-(2,6-difluorophenyl)-5-fluoropicolinamide; 3-amino-M-(4-(3А8, 48, 55)-3-amino-4-hydroxy-5-methylpiperidin-1-yl)pyridin-3-yl)-6-(2,6-difluorophenyl)- 5-fluoropicolinamide; C-amino-

M-(4-(1B8,35)-3-aminocyclohexyl)pyridin-3-yl)-6-(2,6-difluorophenyl)-5-fluoropicolinamide; M-(4-(ZA, 48, 55)-3-amino-4-hydroxy-5-methylpiperidin-1-yl)pyridin-3-yl)-6-(2,6-difluorophenyl)-5-fluoropicolinamide; 3-

amino-M-(4-(ЗВ, 48, 55)-3-amino-4-hydroxy-5-methylpiperidin-1-yl)pyridin-3-yl)-6-(2,6-difluorophenyl)-5- fluoropicolinamide; and M-(4-(3A8, 4B8, 55)-3-amino-4-hydroxy-5o-methylpiperidin-1-yl)pyridin-3-yl)-6-(2,6-difluoro-3-methylphenyl) -5-fluoropicolinamide. Another preferred embodiment relates to a pharmaceutical composition additionally containing an additional agent intended for the treatment of cancer, and the preferred additional agent is selected from the group including irinotecan, topotecan, gemcitabine, 5-fluorouracil, leucovorin, carboplatin, cisplatin, taxanes, tezacitabine, cyclophosphamide , periwinkle alkaloids, imatinib (Gleevec), anthracyclines, rituximab, and trastuzumab.

The compounds of this invention can be used to treat cancers, including hematopoietic malignancies, carcinomas (e.g., lung, liver, pancreas, ovary, thyroid, bladder, or colon), melanomas, myeloid disorders (e.g., myeloid leukemia, multiple myeloma and erythroleukosis), adenomas (for example, villous adenoma of the colon), sarcomas (for example, osteosarcoma), autoimmune diseases, allergic reactions and organ transplant rejection syndromes.

In another object, the present invention relates to the use of a compound of formula I or formula I for the preparation of a medicinal product intended for the treatment of a pathological condition by modulating the activity of proviral integration of Meloni's kinase (RIM kinase). In a preferred embodiment of this object of the present invention, the pathological condition is a cancer selected from the group consisting of carcinoma of the lung, pancreas, thyroid, ovary, bladder, breast, prostate or colon, melanoma, myelogenous leukemia, multiple myeloma and erythroleukosis, villous adenoma of the colon and osteosarcoma.

In another object, the present invention relates to methods of inhibiting the activity of at least one kinase selected from the group consisting of Rit!, Rit2 and Rit3 in a subject or treating a biological pathological condition mediated by at least one of Rit!, Rit2 and Rit3, in a human or animal in need of such treatment, comprising administering to the subject at least one compound of formula I or II in an amount effective to inhibit the kinase in the subject. Therapeutic compounds can be used to treat patients in need of such inhibitors (eg, those suffering from cancers mediated by abnormal serine/greonine kinase receptor signaling).

The term "PIM inhibitor" is used herein to refer to a compound that has an IC value for PIM kinase activity of no more than about 100 µM, and more commonly no more than about 50

MKM obtained in studies of consumption under the influence of RIM, described below in this invention.

The term "alkyl" refers to alkyl groups that do not contain heteroatoms. This term includes straight chain alkyl groups such as methyl, ethyl, propyl, butyl, pentyl, hexyl, heptyl, octyl, nonyl, decyl, undecyl, dodecyl, and the like. The term also includes isomers having branched chain, straight chain alkyl groups, including but not limited to the following, which are exemplified: -CH(CH3)», CH(ISCH3)(CHeCH»3), CH( ISNeaCH»z)2, C(CHz)z, C(CHNaCH»z)z, CHgCH(CHZ)", snesn(CHz3)(SnNesCHzI), SnNegsSnH(CHcCH»z)", SNeb(CHz)zi, CHgS( СНеСН3)з, СсН(СН3СнН(СН3)(СНеСН3),

СснНснсн(СНз), СНАСНСНн(СНИСНегсСН»з»), СНЕСН.Н(СНеСН»)», СНгСНоС(СНЗ3)з, СНСНС(СНСН z) z, сн(Сснза)снес(СНзг:5,, СН(снНЗ3СнН( CHzZISNHn(CHz), CH(CH.HzSnH(CHz)Cn(CHzuCHnesCH"), etc. Thus, the expression alkyl group includes primary alkyl groups, secondary alkyl groups and tertiary alkyl groups. Preferred alkyl groups include alkyl groups that have a linear and branched chain containing from 1 to 12 carbon atoms. A preferred definition of "alkyl" refers to straight-chain C 1 -C 4 -alkyl groups such as methyl, ethyl, n-propyl, and n-butyl. A preferred definition of " alkyl" also includes branched C3-Cv-alkyl groups, including CH(CH3)g2, CHCH(CH3)", CH(CH3)CHnesCnH 5,

C(CHnz)z, CH(ISNHz3ISnN.H»osH», CH(CHZISN(CHz)», SNESN(CHNaiz)SNesSN», SNESNSN(CH Z)», and CH(CHNoCH z)», etc.

The term "alkenyl" means alkyl groups, as defined above, which contain at least one unsaturated moiety, that is, in which two adjacent carbon atoms are linked by a double bond. The term "alkynyl" refers to alkyl groups in which two adjacent carbon atoms are linked by a triple bond. "Alkoxy group" means OK, where K is alkyl.

As used herein, the term "halogen" refers to the chloride, bromide, fluoride, and iodide groups. "Haloalkyl" means an alkyl radical substituted with one or more halogen atoms. Thus, the term "haloalkyl" includes monohaloalkyl, dihaloalkyl, trihaloalkyl, and the like. Typical monohaloalkyl groups include -СНеЕ, -СН2СИ, -СНеСН»еЕ, -

СНеСНеСИ, -СН(Р)СН», -СН(СИ)СН»; typical dihaloalkyl groups include СНСЙИг, -"СНЕ", -СС12СН", -

СН(С)СН2СИ, -СНа.СНСиИ», -СНоСНЕ»; typical trihaloalkyl groups include -C3, -C3, -

ССИ2СНоСИ, -СР2СНоЕ, СН(ИС)СНОИ», -СН(Е)СНЕ» and typical perhaloalkyl groups include -ССизв, -

СЕз, -ССИ2ССиз, -СЕ»2СЕз. "Amino group" in this invention means the group -MN', which can be substituted to form -MAV'.

The term "alkylamino group" as used herein means the group -MEK, wherein K and KE are all independently selected from the group consisting of hydrogen and lower. alkyl The term "arylamino group" in this invention means a group -

ME, in which EC is aryl and E" is hydrogen, below. alkyl or aryl. The term "arylalkylamino group"

in this invention means the group -MKK" in which K is lower arylalkyl and K' is hydrogen, lower alkyl, aryl or lower arylalkyl. The term cyano group means the group -CM. The term nitro group means the group -MO".

"Alkoxyalkyl" refers to the group -alkyl:-O-alkyl, wherein alkyl or alkenyl and alkyl or alkenyl. The term "lower alkoxyalkyl" means alkoxyalkyl, where aiYIK: denotes lower. alkyl or lower. alkenyl and alkenyl means lower. alkyl or lower. alkenyl The term "aryloxyalkyl" means the group - alkyl-o-aryl. The term "arylalkyloxyalkyl" means an alkylenyl-o-arylalkyl group in which arylalkyl means lower. arylalkyl

The term "aminocarbonyl" in this invention means the group -С(0)-МН». "Substituted aminocarbonyl" in the present invention means a C(O)-MEK group in which E is lower. alkyl and E" represents hydrogen or lower alkyl. In some embodiments, K and KE together with the M atom attached to them can form a "heterocycloalkylcarbonyl" group. The term "arylaminocarbonyl" as used herein means a group

C(O)-MAP, in which LC is aryl and EE is hydrogen, is lower. alkyl or aryl. "arylalkylaminocarbonyl" in this invention means the group -C(O0)-MKEK, in which K is lower. arylalkyl and EK denotes hydrogen, lower. alkyl, aryl, or lower. arylalkyl "Aminosulfonyl" in this invention refers to the group -5(0)2-MHg. "Substituted aminosulfonyl" in the present invention means a 5(0)2-MERE group in which E is lower. alkyl and E" represents hydrogen or lower alkyl. The term "arylalkylaminosulfonylaryl" in this invention means the group -aryl-5(0)2-AMN- arylalkyl, in which the arylalkyl is lower. arylalkyl. "Carbonyl" means the divalent group -C (О0). "Carboxy group" means -C(-0)-OH. "Alkoxycarbonyl" means an ester -C(-0)OK, in which K is alkyl. "Lower. "Alkoxycarbonyl" means an ester -C(-0)OK, in which CK is lower alkyl. "Cycloalkylcarbonyl" means -C(-0)-OE, in which E is cycloalkyl. "Cycloalkyl" means mono- or polycyclic, heterocyclic or a carbocyclic alkyl substituent. Carbocycloalkyl groups are cycloalkyl groups in which all ring atoms are carbon atoms. Typical cycloalkyl substituents contain from 3 to 8 atoms in a main chain (ie, ring) in which each main chain atom is a carbon atom or a heteroatom. The term "heterocycloalkyl" in this invention refers to cycloalkyl substituents that contain in the ring structure from 1 to 5 and more often from 1 to 4 heteroatoms. Suitable heteroatoms used in the compounds of this invention are nitrogen, oxygen and typical heterocycloalkyl moieties include, for example, a morpholine group, piperazinyl, opiperidinyl, and the like.

Carbocycloalkyl groups are cycloalkyl groups in which all ring atoms are carbon atoms. When used in connection with cycloalkyl substituents, the term "polycyclic" in this invention refers to fused and unfused cycloalkyl structures. The terms "partially unsaturated cycloalkyl", "partially saturated cycloalkyl" and "cycloalkenyl" all mean a cycloalkyl group which contains at least one unsaturated moiety, i.e. in which two adjacent carbon atoms are linked by a double bond or a triple bond. Illustrative examples include cyclohexenyl, cyclohexynyl, cyclopropenyl, cyclobutynyl, and the like.

The terms "substituted heterocycle" or "heterocyclic group" or "heterocycle" as used herein means any 3- or 4-membered ring containing a heteroatom selected from the group consisting of nitrogen, oxygen, and sulfur, or 5- or a 6b-membered ring containing from 1 to 3 heteroatoms selected from the group consisting of nitrogen, oxygen and sulfur; where the 5-membered ring contains 0-2 double bonds and the b-membered ring contains 0-3 double bonds; where nitrogen and sulfur atoms can optionally be oxidized; where nitrogen and sulfur atoms can optionally be quaternized; and includes any bicyclic group in which each of the above heterocyclic rings is fused to a benzene ring or other 5- or b-membered heterocyclic ring independently defined above. The term "heterocycloalkyl" as used herein means a 5- or b-membered ring containing from 1 to 3 heteroatoms selected from the group consisting of nitrogen, oxygen and sulfur, and the ring does not contain double bonds.

For example, the term heterocyclo-C 1 -alkyl means a b-membered ring containing 5 carbon atoms and a heteroatom such as M. Thus, the term "heterocycle" includes rings in which nitrogen is the heteroatom, as well as partially and fully saturated rings Preferred heterocycles include, for example: diazepinyl, pyryl, pyrrolinyl, pyrrolidinyl, pyrazolyl, pyrazolinyl, pyrazolidinyl, imidazolyl, imidazolinyl, imidazolidinyl, pyridyl, piperidinyl, pyrazinyl, piperazinyl, M-methylpiperazinyl, azetidinyl, M-methylazetidinyl, pyrimidinyl, pyridazinyl, oxazolyl, oxazolidinyl, isoxazolyl, isazolidinyl, morpholinyl, thiazolyl, thiazolidinyl, isothiazolyl, isothiazolidinyl, indolyl, quinolinyl, isoquinolinyl, benzimidazolyl, benzothiazolyl, benzoxazolyl, furyl, thienyl, triazolyl, and benzothienyl.

Heterocyclic fragments can be unsubstituted or monosubstituted or disubstituted by various substituents independently selected from the group consisting of a hydroxy group, a halogen, an oxo group (C-O), an alkylimino group (KM-, in which K represents a lower alkyl or lower alkoxy group), an amino group, an alkylamino group, a dialkylamino group, an acylaminoalkyl group, an alkoxy group, a thioalkyl group, a polyalkyl group, lower. alkyl, cycloalkyl or haloalkyl.

Heterocyclic groups can be attached in various positions, as should be apparent to those skilled in the art of organic and medicinal chemistry in connection with the disclosure of this invention.

Typical heterocyclyls include, for example, imidazolyl, pyridyl, piperazinyl, piperidinyl, azetidinyl, thiazolyl, furanyl, triazolyl benzimidazolyl, benzothiazolyl, benzoxazolyl, quinolinyl, isoquinolinyl, quinazolinyl, quinoxalinyl, phthalazinyl, indolyl, naphthopyridinyl, indazolyl, and quinolizinyl. "Aryl" means optionally substituted monocyclic and polycyclic aromatic groups containing in the main chain from 3 to 14 carbon atoms or heteroatoms and includes carbocyclic aryl groups and heterocyclic aryl groups. Carbocyclic aryl groups are aryl groups in which all ring atoms of the aromatic ring are carbon atoms. The term "heteroaryl" in this invention means aryl groups containing from 1 to 4 heteroatoms as ring atoms of the aromatic ring, and the other ring atoms are carbon atoms. When used in connection with aryl substituents, the term "polycyclic aryl" herein refers to fused and unfused ring structures in which at least one ring structure is aromatic, such as, for example, a benzodioxazole group (which contains a heterocyclic structure fused to a phenyl group , that is, naphthyl, etc.

Typical aryl moieties used as substituents in the compounds of this invention include phenyl, pyridyl, pyrimidinyl, thiazolyl, indolyl, imidazolyl, oxadiazolyl, tetrazolyl, pyrazinyl, triazolyl, thiophenyl, furanyl, quinolinyl, purinyl, naphthyl, benzothiazolyl, benzopyridyl and benzimidazolyl, etc. "Optionally substituted" or "substituted" means the replacement of one or more hydrogen atoms by a monovalent or divalent radical. Suitable substituting groups include, for example, hydroxy, nitro, amino, imino, cyano, halogen, thio, sulfonyl, thioamide, amidine, imidino, oxo, oxamidine, methoxamidine, imidino, guanidine, sulfonamide, carboxy group, formyl, lower. alkyl, halogen - lower. alkyl, lower. alkylamino group, halogen-lower. alkylamino group, lower. alkoxy group, halogen-lower. an alkoxy group, lower. alkoxyalkyl, alkylcarbonyl, aminocarbonyl, arylcarbonyl, arylalkylcarbonyl, heteroarylcarbonyl, heteroarylalkylcarbonyl, alkylthio, aminoalkyl, cyanoalkyl, aryl, etc.

A substituting group can itself be substituted. The substituent of the substituting group can be a carboxy group, halogen, nitro group, amino group, cyano group, hydroxy group, lower. alkyl, lower. alkoxy group, aminocarbonyl, -5E, thioamide group, -5OzH, -5O2K or cycloalkyl, where K is usually hydrogen, hydroxy or lower. alkyl

If the substituted substituent includes a group having a linear chain, then the substitution can occur within the chain (for example, 2-hydroxypropyl, 2-aminobutyl, etc.) or at the end of the chain (for example, 2-hydroxyethyl, 3-cyanopropyl, etc. .). Substituted substituents can be a branched or cyclic group of covalently bonded carbon atoms or heteroatoms having a linear chain. It should be understood that the above definitions do not include unacceptable substitution patterns (eg, methyl substituted with 5 fluoride groups, or a halogen atom substituted with another halogen atom). Such unacceptable substitution schemes are well known to those skilled in the art.

It should be understood by those skilled in the art that the compounds of this invention, or stereoisomers thereof, as well as pharmaceutically acceptable salts, esters, metabolites and prodrugs of each, may undergo tautomeric transformations and therefore may exist in various tautomeric forms, in in which the proton of one atom of the molecule moves to another atom and the chemical bonds between the atoms of the molecule are rearranged accordingly. See, for example, Magsi, Admapsei Ogdapis

Spetivzigu: VNeasiiopv5, Mespapiztv apa zygisiigev, Boshpii Ediiop, dopp U/ieu b 5op5, rade5 69-74 (1992).

As used herein, the term "tautomer" refers to compounds formed by proton transfer, and it is to be understood that all tautomeric forms, if they may exist, are within the scope of the present invention.

The compounds proposed in this invention, as well as pharmaceutically acceptable salts, esters, metabolites and prodrugs of each of them, may contain asymmetrically substituted carbon atoms. Such asymmetrically substituted carbon atoms can lead to the compounds proposed in this invention existing in enantiomeric, diastereoisomeric and other stereoisomeric forms, which can be designated as (K)- or (5)-forms by means of designations of absolute stereochemical configurations. Therefore, all such possible isomers, individual stereoisomers in their optically pure forms, their mixtures, racemic mixtures (or "racemates"), mixtures of diastereoisomers, as well as individual diastereoisomers of the compounds proposed in this invention are included in the scope of the present invention. The terms "5" and "KE" configuration when used in this invention have the meanings defined in the publication ISRAS 1974 Kesottepaaiyop5 og besiyop E,

Appdatenpia! 5iegeospetivigh, Rita Arry. Spent 45:13-30 (1976). Notations са and Б are used to determine the positions of rings in cyclic compounds. "-The position in the plane in question is the position in which the best substituent is located in the position with the lower number. Substituents located on the opposite side of the considered plane are designated by the symbol B. It should be noted that these designations differ from the designations of cyclic stereoisomers, for which "a" means "below the plane" and denotes the absolute configuration. The terms a- and VD-configuration when used in this invention have the meanings defined in the publication of Spetis! Arzigashi5 Ipdeh Scide-Arrepaikh IM (1987)

ragadgari 203.

As used herein, the term "pharmaceutically acceptable salts" means non-toxic acid salts or non-toxic alkaline earth metal salts of compounds of formulas I or II. These salts can be obtained by final isolation and purification of compounds of formulas I or II or by separate reactions of basic or acidic groups with a suitable organic or inorganic acid or base, respectively. Exemplary salts include, but are not limited to, the following: acetate, adipate, alginate, citrate, aspartate, benzoate, benzenesulfonate, bisulfate, butyrate, camphorate, camphorsulfonate, digluconate, cyclopentanepropionate, dodecylsulfate, ethanesulfonate, glucoheptanoate, glycerophosphate, hemisulfate, heptanoate, hexanoate, fumarate, hydrochloride, hydrobromide, hydroiodide, 2-hydroxyethanesulfonate, lactate, maleate, methanesulfonate, nicotinate, 2-naphthalene sulfonate, oxalate, pamoate, pectinate, persulfate, Z-phenylpropionate, picrate, pivalate, propionate, succinate, sulfate, tartrate, thiocyanate, p-toluenesulfonate and undecanoate. In addition, nitrogen-containing main groups can be quaternized with reagents such as below. alkyl halides, such as methyl-, ethyl-, propyl- and butyl chlorides, -bromides and -iodides; dialkyl sulfates, such as dimethyl, diethyl, dibutyl, and diamyl sulfates, long-chain halides, such as decyl, lauryl, myristyl, and stearyl chlorides, -bromides and -iodides, arylalkyl halides, such as benzyl and phenethyl bromides, etc. In this way, products that dissolve or disperse in water or oil are obtained.

Examples of acids that can be used to prepare pharmaceutically acceptable acid addition salts include inorganic acids such as hydrochloric acid, nitric acid, sulfuric acid and phosphoric acid, and organic acids such as oxalic acid, maleic acid, methanesulfonic acid, succinic acid and citric acid. Addition salts with bases can be obtained by final isolation and purification of compounds of formula (I) or by separate reactions of acid groups of carboxylic acids with a suitable base, such as the hydroxide, carbonate or bicarbonate of a pharmaceutically acceptable metal cation, or with ammonia or with a primary, secondary or a tertiary organic amine. Pharmaceutically acceptable salts include, but are not limited to, salts containing alkali and alkaline earth metal cations, such as sodium, lithium, potassium, calcium, magnesium, aluminum, and the like. as well as non-toxic ammonium, quaternary ammonium and amine salts, including but not limited to ammonium, tetramethylammonium, tetraethylammonium, methylamine, dimethylamine, trimethylamine, triethylamine, ethylamine, etc. Other typical organic amines used to prepare base addition salts include diethylamine, ethylenediamine, ethanolamine, diethanolamine, piperazine, and the like.

As used herein, the term "Pharmaceutically acceptable ester" refers to esters that are hydrolyzed by ip mimo and include those that are readily degraded in the human body to release the parent compound or a salt thereof. Suitable ester groups include, for example, those formed from pharmaceutically acceptable aliphatic carboxylic acids, preferably alkanecarboxylic, alkenecarboxylic, cycloalkanecarboxylic, and alkanedicarboxylic, in which each alkyl or alkenyl moiety preferably contains no more than 6 carbon atoms. Typical examples of preferred esters include, but are not limited to, formates, acetates, propionates, butyrates, acrylates, and ethyl succinates.

The term "pharmaceutically acceptable prodrugs" when used in this invention means such prodrugs of the compounds proposed in this invention, which from a medical point of view can be used to interact with the tissues of humans and lower animals without the manifestation of undesirable toxicity, irritation, allergic reactions, etc. , are characterized by a reasonable benefit/risk ratio and are effective for their intended use, as well as, when possible, the zwitterionic forms of the compounds proposed in this invention. The term "prodrugs" refers to compounds that rapidly undergo conversion to ip mimo and form the original compounds of the above formula, for example, by hydrolysis in the blood. A detailed discussion is given in the publications of T. Nidispi apa M. 5ieiYiia, Rgo-dpyde5 az Momei! OyeiYimegu Zuzietv, Moi. 14 "EE She A.S.5. 5utrovzisht zZegie5, and Edulaga V. Kospe, ea., Viohemegvirie Saigieg5 ip Ogyd YuOezidp,

Ategisap RIPagtasetschisa! Avzosiayop apa Regdatop Rge55, 1987, which are incorporated herein by reference.

Any formula given in this invention also characterizes unlabeled forms as well as isotopically-labeled forms of compounds. Isotopically labeled compounds have the structures described by the formulas given in this invention, except that one or more atoms are replaced by atoms having the atomic masses or mass numbers indicated. Examples of isotopes that can be incorporated into the compounds of this invention include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorus, sulfur, fluorine, and chlorine, such as 2H, ЗН, "C, 7196, 190, 19M, 8tr, z2p, 855, 360, 125|, respectively. The scope of the present invention includes various isotopically-labeled compounds proposed in this invention, for example, which include radioactive isotopes such as ZN, 7zC, and 1460. Such isotopically-labeled compounds can be used to study metabolism (preferably containing 77), reaction kinetics (containing, for example, hN or ZN), in detection and imaging methodologies such as positron emission tomography (PET) or single photon emission computed tomography (SPECT), including studies distribution of the drug or substrate in tissues or radiation therapy of patients.

In particular, a compound containing or labeled with E may be particularly preferred for PET or SPECT studies. The isotopically labeled compounds of the present invention and their prodrugs can generally be prepared by methods disclosed in the Schemes or Examples and syntheses described below by replacing the isotope-free reagent with a readily available isotope-labeled reagent.

In addition, substitution with heavier isotopes such as deuterium (i.e. 2H or 0) may provide some therapeutic advantages due to their higher metabolic stability, for example, an increased half-life of ip mimo or the possibility of using lower doses, or an improvement in the therapeutic index. It should be understood that in this context deuterium is considered as a substituent in the compound of formula (I). The concentration of such a heavier isotope, in particular, deuterium, can be determined using the isotopic enrichment factor. The term "isotope enrichment factor" when used in this invention means the ratio of the content of the isotope to the content of a particular isotope in nature. If the substituent in the compound proposed in this invention means deuterium, then such a compound is characterized by an isotopic enrichment factor for each labeled deuterium atom equal to at least 3500 (the deuterium content for each labeled deuterium atom is equal to 52.5 95), not less than 4000 (deuterium content equal to 60 95), not less than 4500 (deuterium content equal to 67.5 95), not less than 5000 (deuterium content equal to 75 95), not less than 5500 (deuterium content equal to 82.5 95), not less than 6000 (content deuterium equal to 90 95), not less than 6333.3 (deuterium content equal to 95 9), not less than 6466.7 (deuterium content equal to 97 95), not less than 6600 (deuterium content equal to 99 95) or not less than 6633.3 (content deuterium equal to 99.5 b).

Isotopically labeled compounds of formula (I) can usually be prepared by conventional methods known to those skilled in the art, or by methods similar to those described in the appended examples and syntheses using an isotopically labeled reagent instead of the unlabeled reagent previously used.

It should be apparent to those skilled in the art that the compounds of this invention, or tautomers, prodrugs, and stereoisomers thereof, as well as pharmaceutically acceptable salts, esters, and prodrugs of each, may undergo further transformations by metabolism in the human body or animal or in the cell with the formation of metabolites. The term "metabolite" as used herein means any derivative formed in a subject after administration of the parent compound. Derivatives can be formed from the parent compound by various biochemical transformations occurring in the subject, such as, for example, oxidation, reduction, hydrolysis or conjugation, and include, for example, oxides and demethylated derivatives. Metabolites of a compound of the present invention can be identified by standard techniques known in the art. See, for example, Vepoiipi, ch. ei ai., U. Med. Spent 40:2011-2016 (1997); Znap, 0. ei ai., 9). RNnagtt. 5 86(7):765-767; Vad5pame K., Ogyd Oem. Nev. 34:220-230 (1995); Vodog, M., Admapsez and Ogyd Nev. 13:224-331 (1984); Vypddaat, N., Oyevidp oi Rgodgtzoz5 (EiIbzemieg Rge55 1985); and I agzep, I. K., Oezidp apa Arriisaeop oi Rgodgydz, Ogia Oezidp apa OyemeIortepi (Ktodzdaaga-i agzep sei a!., ed. 5., Naglosy Asadetis RybiivNetgv, 1991). It should be understood that individual chemical compounds, which are metabolites of compounds of formulas II or II, or their tautomers, prodrugs and stereoisomers, as well as pharmaceutically acceptable salts, esters and prodrugs of each of them are included in the scope of the present invention.

The term "cancer" refers to cancers that can be effectively treated by inhibition of Rit kinase, including, for example, solid cancers such as carcinomas (eg, lung, pancreas, thyroid, ovary, bladder, breast, prostate, or colon), melanomas, myeloid disorders (eg, myeloid leukemia, multiple myeloma, and erythroleukemia), adenomas (eg, villous adenoma of the colon), and sarcomas (eg, osteosarcoma).

Methods of synthesis

The compounds of this invention can be prepared by methods known to those skilled in the art. For example, as shown in the scheme, cyclohexanediones through monotriflates can be converted into the corresponding esters of cyclohexenoneboronic acid, which, under palladium catalysis, form a bond with the carbon atom of 4-chloro, 3-nitropyridine to obtain nitropyridine-substituted cyclohexenones I. Reduction of the enone group can give cyclohexenol II , which after protection of the hydroxy group, reduction of the nitro group and alkene, reaction of the combination with the formation of amide and removal of the protecting group can give cyclohexanolamides PI. Cyclohexenol I can also be introduced into the reaction

Mitsunobu with phthalimide and obtain protected aminocyclohexene IM. After the reduction of the nitro group and the alkene aminocyclohexylpyridylaniline Ma containing a phthalimide protecting group, it is possible to react the combination with the formation of an amide and remove the protecting group and obtain aminocyclohexanamides

E. The corresponding aminocyclohexanepyridylaniline MO, containing the Bos protective group, can also be obtained from cyclohexenol II in the following way: protect the hydroxy group, reduce the alkene and nitro group, introduce the CbB7 protective group into the pyridylamine, remove the silyl protective group, carry out oxidation by

Dessom-Martin in cyclohexanone, carry out reductive amination with benzylamine, remove protective groups Сб? and Vp and remove the Bos protecting group from the primary aliphatic amine. In the case of amides

ШІ and Mi, if K2 denotes a halogen or triflate, then the amides PSh and MI can be additionally modified according to standard modification techniques by introducing substituted aryl, alkyl, and heteroaryl by

K2. For example, if Ka2 denotes Bg, various modifications of K2 can be performed by reaction with boronic acids or organometallic reagents or by conversion into the corresponding boronic acid ester and reaction with aryl/heteroaryl halides or triflates.

Scheme 1. : i si V ische in Nasoya in kole, Ra arri si in Su cho, ;

Executioner; schi BO Kati to dioxai VO "During Var and | and o

Sle ru oh nya -- ---- shi Ko shshnhhu . only MO p ote vov 0 gcharROS', | nya disvsanisy MN azs macho I mavn Code ee Aa oo V an Kai "YAR Cat. Me sesiat NGO Re; iVAYU TT Gu

EN, what. I ftapimid sce Na Ros; aso N t kilk, Vshad Ko Tge pes bu bu shkh HT - і: tvemiosi, Sh 1.tvomsi, M sho imidazol, DIYAf ltidahopo, DIMF 2. In FIG. Man. 2: On RKS, MeON 1: lions; NOAT, EDOE, DM

Pitch, notes, / Z. colors, DMA, DAM | 2. Nam Na; MeOH,

ED DM, Zbtod. / 4 NS EYUN | vkas 4. NOSI, EMIN WE! 8. Redgent of Des-Martin, a copy of V. Vomna, ChVN, kh t. in: horse Va and ey Ke M Ka MNVos Katoret and MNa Ka . Come on! : NO: t i lagi t-sh iv) КБХДАСНТ з и в, ба и чно выше рол у 0

NNYA M and p

PN NN zbo INN. In cha ELENA; E or NN

Alternatively, as shown in Scheme 2, cyclohexenol II can be dehydrated to obtain cyclohexadiene, which after epoxidation (through the formation of bromohydrin and cleavage of HBg or directly from MHPBK (m-chloroperoxybenzoic acid)) and opening of the epoxide ring with azide gives cyclohexenyl azido alcohol MI. Cyclohexenyl azido alcohol MI can be converted into trans-protected aminohydroxyaniline Ma by reduction of the azide group, protection of the hydroxy group, and reduction of the alkene and nitro group. Alternatively, the cyclohexenyl azido alcohol MI can be converted into a cis-protected aminohydroxyaniline Mr by reduction of the azide group and introduction of the Bos protecting group, mesylation of the alcohol and intramolecular cyclization into a cis-cyclic carbamate, subsequent introduction of the Bos protecting group and reduction of the alkene and nitro group. Obtained cyclohexylpyridylanilines

Mia and MIIB can be converted into the corresponding pyridinamides MPa and MIB by means of a combination reaction with the formation of an amide, cleavage of an acetate or cyclic carbamate, and removal of the Bos protecting group.

If K2 denotes a halogen or a triflate, then the amides MISHa and MB can be further modified according to standard modification techniques by introducing substituted aryls, alkyls, and heteroaryls by

K2 after the formation of an amide bond and until the complete removal of protective groups. For example, if K2 denotes B, then various modifications of K2 can be performed by reaction with boronic acids or organometallic reagents or by conversion into the corresponding boronic acid ester and reaction with aryl/heteroaryl halides or triflates.

Scheme 2.

Ka v v Ve Esh ay

Kaya on Na syu, Va ton shk

A ptsk S. ve; Nai T a and dioxane, 10 s Her tgF KONVy, TGf for room three

I. I And - What t

M M in Iyai district

Ii z МХПК:ДХМ d-t di ANNA

Ode he

And "MNVoye ku Ke."

TRU that 11 Acid; NOAT, ED, DM shchi Ka 2 bossa t gi svasOz, Mas EN nu v. HE vkso le eMNa. ZI 25'yaTFKSNaS and che

Vako "z I Na ras scheche neni: an y "la mita VEN, og Na von u 1) eMe i dn t zo vuVOSO du "Vas va ho Na

IM code all tek n 1) Acid, noltkdyu blows pe in pn n grvi, Men i Ghvsnnai you

FPiridnk ps 007 mn. Зо5жЖ ТЕКИсНАС В щ ее 00 ГТ яна ао тн зр в

MV KAN, or MN.

IN

Alternatively, as shown in scheme C, it is possible to obtain trisubstituted 5-alkyl-4-hydroxy-3-aminopiperidines and modify them to obtain trisubstituted 5-alkyl-4-hydroxy-3-aminopiperidinylpyridinamides IX in this way. The Garner reaction of aldehyde with (2)-4-benzyl-3-propionyloxazolidin-2-one, followed by the introduction of the protective group TV5 into the resulting alcohol gives compound X. Reduction of oxazolidinone, followed by the addition of the azide group gives the intermediate product XI. Removal of the protective group in an acidic environment gives the corresponding amino alcohol, which after the introduction of the protective group Bos and the subsequent mesylation of the primary alcohol gives the intermediate product XII. Reduction of the azide allows to obtain piperidine, which is then introduced into the reaction with 4-chloro-3-nitropyridine, reduced to the amine, introduced into the reaction of the combination with the corresponding carboxylic acid, and the protective group is removed, and trisubstituted 5-methyl-4-hydroxy-3-aminopiperidinylpliridinamides are obtained THEIR. If K1 denotes a halogen or a triflate, then the amide IH can be additionally modified according to standard modification methods by introducing substituted aryls, alkyls and heteroaryls by K1! after the formation of an amide bond and until the complete removal of protective groups. For example, if K1! denotes Bg, then various modifications of K1 can be performed by reaction with boronic acids or organometallic reagents or by conversion into the corresponding boronic acid ester and reaction with aryl/heteroaryl halides or triflates.

Scheme 3.

to a 13 fast; diZA a a stvo Z SHVN,, ECON, t, lis

Ku nu dam, as Hr from OS to so in rus o V pot no) Vosia pot Vosv y

What voasr 21 TE5O TE, lutidin t. p. 7) DYAD, ERI and - more dhmovs I DFFATG ! ha SU x 1ZEKON, PTS kvyatnya with: an reverse holodipenixite and es, diIZA, Men Nam. with Wasp; DYZA, DHM Sto a chloro-3-nitropyridin Sty v. cha, RY nt to DYZA, ERUZN. VSS vl G and 7 DMAN, OS MNNvo z Ra/s, men g u

ZHI "and Kispota, ED, nal Ddyaf not me

YAM NSHE doksani A KI1- NE or MN

Their

Alternatively, as shown in Scheme 4, it is possible to obtain trisubstituted 5-methyl-4-hydroxy-3-aminopiperidines and modify them to obtain trisubstituted 5-methyl-4-hydroxy-3-aminopiperidinyl amides Xi in this way. The reaction of crotyl ethers of boronic acid with aldehyde 5egoVp, the subsequent formation of a cyclic carbamate, oxidative cleavage of the alkene and reduction gives the compound KHIM containing a hydroxy group. Removal of the benzyl protecting group, subsequent bistosylation and reaction with p-methoxybenzylamine, and removal of the amine protecting group gives piperidine ХУ. The reaction of substituted piperidine XM with halonitrosubstituted arenes or heteroarenes, subsequent introduction of a carbamate protecting group, reduction of a nitro group, combination reaction with the formation of an amide, opening of a cyclic carbamate, and removal of the protecting group gives trisubstituted 5-methyl-4-hydroxy-3-aminopiperidinylamides ХИЙ. If KZ denotes a halogen or triflate, then the amide XM can be further modified according to standard modification techniques by introducing substituted aryls, alkyls, and heteroaryls by KZ. For example, if KZ denotes Bg, then various modifications of KZ can be performed by reaction with boronic acids or organometallic reagents or by conversion into the corresponding ester of boronic acid and reaction with aryl/heteroaryl halides or triflates.

Scheme 4.

Va SNI, Mon 5th G ts-i o«MNVos ! ry Man, li F a

In T sa - ! - in I ova ovo OV and she . Shchy) o I. Na Raiu Men o - 1. Mag OvSuibmolu 00-50 8 PTO Prydnn Vus

MON, SCHO ts ge 3. MNORMV, DE A iy

M

2: Mavn,, EN NS no ov MR. TOM, and Hor. N bob" stage A nkh and "Konni: and kt; MON, 2 h. hu a a- in 1. 1055 RAS, On MON on

Ia. NeBSI DIKASVS. . | hey Not in Vosa, DMAP m 8: YSpota, NOBT, EDH, DF |. and -- | ("F 4 l f f 5 ' - - - -- "k still MO, f M n pa o yar

M Za Svsoz, MeOon And Her dove

ZOM NO shokoan M x.

The compounds of this invention can be used ip miigo or ip mimo to suppress the growth of cancer cells. The compound may be used alone or in compositions with a pharmaceutically acceptable carrier or inert excipient. Suitable pharmaceutically acceptable carriers or inert excipients include, for example, processing additives and substances that modify and improve drug delivery, such as, for example, calcium phosphate, magnesium stearate, talc, monosaccharides, disaccharides, starch, gelatin, cellulose, methylcellulose, sodium carboxymethylcellulose salt, dextrose, hydroxypropyl-B-cyclodextrin, polyvinylpyrrolidinone, low melting point waxes, ion exchange resins, etc., as well as combinations of any two or more of them. Other suitable pharmaceutically acceptable inert excipients are described in the publication Ketipdiopye Rpaptaseshisa! Zsiepse5, Musk Ryb. So., Mem degheu (1991), which is incorporated herein by reference.

Effective amounts of the compounds of this invention will generally be any amount sufficient to inhibit detectable Rit activity as determined by each of the assays described herein, by other assays for Rit kinase activity known to those skilled in the art. general training in this field of technology, or such as is found by suppressing or weakening the symptoms of cancer. The amount of active ingredient that can be combined with carriers to form a single dosage form will vary depending on the recipient being treated and the particular route of administration. It should be understood, however, that the exact dose for any particular patient will depend on many factors, including the activity of the particular compound used, age, body weight, general health, sex, diet, time of administration, route of administration, rate of elimination , the combination of drugs and the weight of the specific disease being treated. In each case, the therapeutically effective amount can be easily determined with the help of standard tests and the decision is made by the doctor based on his professional training.

For the purposes of the present invention, a therapeutically effective dose is usually a full daily dose administered to the recipient as a single or divided doses in amounts that may range, for example, from 0.001 to 1000 mg/(kg body weight) per day, and even better from 1 .0 to 30 mg/kg of body weight) per day. A single dose of the composition can be a fraction of such amounts, so that in total they form a daily dose.

The compounds of this invention may be administered orally, parenterally, sublingually, by aerosol or inhalation spray, rectally, or topically in the form of single dose compositions optionally containing conventional non-toxic pharmaceutically acceptable carriers,

excipients and solvents. Topical administration may also involve the use of transdermal means such as transdermal patches or iontophoresis equipment. The term "parenteral" as used herein includes subcutaneous injections, intravenous, intramuscular, epigastric injections and infusions.

Preparations for injection, for example, sterile aqueous or oily suspensions for injection can be prepared using suitable dispersing or wetting agents and suspending agents known in the art. A sterile preparation for injection can also be a sterile solution, suspension or emulsion for injection in a non-toxic parenterally acceptable diluent or solvent, for example, a solution in 1,3-propanediol. Acceptable carriers and diluents that may be used include water, Ringer's solution, and isotonic sodium chloride solution. In addition, sterile non-volatile oils are usually used as a solvent or suspending medium. Any liquid fixed oil can be used for this purpose, including synthetic mono- or diglycerides. In addition, fatty acids such as oleic acid are used to prepare injectables.

Suppositories for rectal administration of the drug can be prepared by mixing the drug with a suitable inert, non-irritating excipient, such as cocoa butter or polyethylene glycols, which are solid at ambient temperature but liquid at rectal temperature and therefore melt in the rectum. and release the drug.

Solid dosage forms for oral administration may include capsules, tablets, pills, powders, and granules. In such solid dosage forms, the active compound can be mixed with at least one inert diluent such as sucrose, lactose or starch. Such solid dosage forms may include, as is common practice, additional substances that are not inert diluents, such as lubricating agents such as magnesium stearate. In the case of capsules, tablets and pills, dosage forms may also include buffering agents. Tablets and pills can also be made with enteric coatings. Liquid dosage forms for oral administration may include pharmaceutically acceptable emulsions, solutions, suspensions, syrups and elixirs containing inert diluents commonly used in the art, such as water. Such compositions may also contain auxiliary substances, such as wetting agents, emulsifying and suspending agents, cyclodextrins and sweeteners, flavors and aromas.

The compounds of the present invention can also be administered in the form of liposomes. As known in the art, liposomes are usually derived from phospholipids or other lipids. Liposomes are formed by mono- or multi-layered hydrated liquid crystals that are dispersed in an aqueous medium. Any non-toxic, physiologically acceptable, metabolizable lipid capable of forming liposomes can be used. The compositions proposed in this invention, in liposomal form, in addition to the compound proposed in this invention, may contain stabilizers, preservatives, inert fillers, and the like. The best lipids are phospholipids and phosphatidylcholines (lecithins), natural and synthetic. Methods for preparing liposomes are known in the art. See, e.g., the publication of Rgezsoi, Ea., Meijoa5 ip Seii Viiodou, Moishte KHIM, Asadetis Rgev55, Mem/ Mogk,

M.MU., Z3Z vy zed. (1976).

Although the compounds of this invention can be administered as a single drug, they can also be used in combination with one or more other agents used to treat cancer. The compounds proposed in this invention can also be used in combination with known drugs and anticancer agents, and combinations of the compounds proposed in this invention with other anticancer agents or chemotherapeutic agents are within the scope of the present invention. Examples of such tools are given in the publication Sapseg Rgipsirie5 apa Rgasiise oi

Opsoiodu, U. T. Oemiga apa 5. NeiItap (eayogv), bii eyiop (Reb. 15, 2001), Ghirripsoii MUShatv 8 UMiiYKip5

Rybiyhpeg5. One of ordinary skill in the art should be able to determine which combinations of agents will be used according to the characteristics of the particular drugs and those undergoing cancer treatment. Such anticancer agents include, but are not limited to, the following: estrogen receptor modulators, androgen receptor modulators, retinoid receptor modulators, cytotoxic/cytostatic agents, antiproliferative agents, prenylprotein transferase inhibitors, HMO-CoA reductase inhibitors and other angiogenesis inhibitors, cell proliferation inhibitors, and viability signaling agents, agents that induce apoptosis, and agents that affect key stages of the cell cycle. The compounds of this invention may also be used in conjunction with radiation therapy.

Therefore, in one embodiment of the present invention, the compounds of the present invention are also used in combination with known anticancer agents, including, for example, estrogen receptor modulators, androgen receptor modulators, retinoid receptor modulators, cytotoxic agents, antiproliferative agents, prenylprotein transferase inhibitors, NMO-CoA reductases, HIV (human immunodeficiency virus) protease inhibitors, reverse transcriptase inhibitors and other angiogenesis inhibitors.

In some currently preferred embodiments of the present invention, typical agents that can be used in combination with the compounds of this invention for the treatment of cancer include, for example, irinotecan, topotecan, gemcitabine, 5-fluorouracil, leucovorin, carboplatin, cisplatin, taxanes, tezacitabine , cyclophosphamide, periwinkle alkaloids, imatinib (Gleevec), anthracyclines, rituximab, trastuzumab, as well as anticancer chemotherapy agents.

The above compounds used in combination with the compounds of the present invention can be used in therapeutic amounts as described in the publication Rpuzisiape' rezk Ketiegepse (ROC) 471p Eyop (1993), which is incorporated herein by reference, or in such therapeutically applicable amounts as would be known to one of ordinary skill in the art.

The compounds of this invention and other anticancer agents can be administered at the recommended maximum clinical dosage or at lower doses. The dosage amounts of the active compounds in the compositions proposed in this invention can be varied to provide the desired therapeutic effect according to the route of administration, the severity of the disease and the response of the patient. The combination can be administered in the form of separate compositions or in the form of a single dosage form containing both agents. When administered in the form of a combination, the drugs can be prepared in the form of separate compositions that are administered simultaneously or at different points in time, or the drugs can be administered in the form of a single composition.

In one embodiment, the present invention relates to a method of inhibiting Rit!, Rit2 or

RitZ3 in humans or animals. The method includes administering to a subject in need of it a compound or a pharmaceutically acceptable salt thereof, which corresponds to each of the embodiments of the compounds of formula I or

II, in an effective amount.

The present invention will be more easily understood using the following examples, which are presented for illustration only and are not intended to limit the present invention.

Examples

In the following examples, the compounds corresponding to the preferred embodiments were synthesized according to the methods described in this invention or according to other methods known in the art.

Compounds and/or intermediates were characterized by high-performance liquid chromatography (HPLC) using an Umayeg5 MiShepisht chromatographic system with a separation modulus of 2695 (Lightning, MA). Analytical columns with reversed phase Rpepotepeh Hypa C18-5 μm, 4.6x50 mm, manufactured by AiShesp (Oyeepieia, I) were used. Elution was used in a gradient mode (flow rate 2.5 ml/min), usually starting from a mixture of 5 95 acetonitrile/95 956 water and ending with 100 95 acetonitrile, within 10 min. All solvents contained 0.1 96 trifluoroacetic acid (TFA). The compound was detected by absorption in the ultraviolet (UV) region of the spectrum at a wavelength of 220 or 254 nm. We used solvents for HPLC produced by the companies VygaiskK apa daskzop (MizKedap, MI) or Rihpeg Zsiepiyis (RINbBrygoi, RA).

In some cases, purity was determined by thin-layer chromatography (TLC) using silica gel-coated glass or plastic plates, such as flexible Wackeg-Rieh 5iis Se! 182-R. TLC data were easily obtained by visual detection under ultraviolet light or by using the well-known iodine vapor staining technique or various other staining techniques.

Mass spectrometric studies were performed on one of three instruments for LCMS (liquid chromatography-mass spectroscopy): MUaseg5 Zuziet (AyShapse NT NR'S and Misgotav55 20) mass spectrometer; column: Esiirse HOV-S18, 2.1x50 mm; gradient mode: 5-95 95 (or 35-95 95, or 65-95 95 or 95-95 95) acetonitrile in water with the addition of 0.05 95 TFA for 4 min; flow rate 0.8 ml/min; molecular weight range 200-1500; potential difference on the cone is 20 V; column temperature 40 C), another

Umayet5 Zuziet (ASSOSHITM ORI S 5uziet and 20 2000 5zuziet; column: ASOSHITU ORI S NZZ-S18, 1.8 μm, 2.1x50 mm; gradient mode: 5-95 95 (or 35-95 95, or 65-95 95 or 95-95 95) acetonitrile in water with the addition of 0.05 95 TFC for 1.3 min; flow rate 1.2 ml/min; molecular weight range 150-850; potentials on the cone 20 V; column temperature 50 "С) or Neulei RasKaga Zuyeiet (Zegie5 1100

NRIS; column: Esiirse HOV-S18, 2.1x50 mm; gradient mode: 5-95 95 acetonitrile in water with the addition of 0.05 95 TFC for 4 min; flow rate 0.8 ml/min; molecular weight range 150-850; the potential difference on the cone is 50 V; the temperature of the column is 30 "C). All given masses refer to protonated initial ions.

Nuclear magnetic resonance (NMR) studies were performed for some compounds on a Magiap 400 MHz NMR instrument (Raio Akyo, SA). TMS (trimethylsilane) or a known chemical shift of the solvent was used as a standard.

Preparative separations were carried out using a Riazp 40 chromatographic system by Kroii, OA (Vioiade, Spapoitsevmije, MA) or by flash column chromatography using silica gel (230-400 mesh) as a packing, or by HPLC using Umayeg5 2767 Zatrie

Mapadeg, C-18 columns with reversed phase, 30x50 mm, flow rate 75 ml/min. Typical solvents used for the RiIazp 40 Violiade system and flash column chromatography were dichloromethane, methanol, ethyl acetate, hexane, acetone, aqueous ammonia (or ammonium hydroxide), and triethylamine. Typical solvents used for reversed-phase HPLC were acetonitrile at various concentrations and water supplemented with 0.1 95 trifluoroacetic acid.

It should be understood that the organic compounds of the preferred embodiments may have tautomerism. Since the chemical structures in this application may represent only one of the possible tautomeric forms, it should be understood that preferred embodiments include any tautomeric form of the depicted structure.

It should be understood that the present invention is not limited to the variants of implementation presented in this invention for illustration, but includes all forms thereof corresponding to the scope of the above description.

In the examples below and throughout this application, the following abbreviations have the meanings indicated. Terms for which definitions are not given have generally accepted meanings.

Abbreviations

DMF 0 |M.oM-dimethylformamide.7/:/ o)

DFFFO 0/1 -bis(idiphenylphosphino)ferrocene ethyl carbodiimide hydrochloride tris(dibenzylideneacetone)dipalladium(0) tetrakis(triphenylphosphine)palladium (0) adduct of dichloro-(1,2- bisidiphenylphosphino)ethane)-palladium) with dichloromethane

Synthesis of 3-oxocyclohex-1-enyltrifluoromethanesulfonate in dO

OT

To a solution of cyclohexane-1,3-dione (1 equiv.) in DCM (0.4 M) was added MagSO3 (1.0 equiv.) and cooled to 0 °C dropwise over 1 h at room temperature under a nitrogen atmosphere 1120 (1.0 eq.) in DCM (5 M) was added. After the addition, the reaction mixture was stirred for 2 h (dark red solution). The solution was filtered and a saturated solution of MansSoO2 was added to the filtrate (carefully), then the organics were extracted, washed with brine, then dried over Magn5Ox and concentrated.

The crude substance was used in the next stage without additional purification. 3-oxocyclohex-1-enyl trifluoromethanesulfonate was obtained in a yield of 67 95. Triflate decomposes during storage and must be used immediately in the next reaction. РХ/МСОС-244.9/286.0 (MAN and

MASNZSM); KI-O, 88 min.

Synthesis of 3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl cyclohex-2-enone in

IN.

AH

To a solution of 3-oxocyclohex-1-enyltrifluoromethanesulfonate (1.0 equiv.) in degassed dioxane (0.3

M) bis(pinacolato)diboron (2.0 equiv.), COAc (3.0 equiv.) and Ra(arr)Si»-DCM (0.05 equiv.) were added. The reaction mixture was heated at 80 "C for 2 hours, then filtered. The solution in dioxane was used at the next stage without additional purification. LC/MS - 140.9 (MAN of boronic acid).

Synthesis of 3-(3-nitropyridin-4-yl)ucycloheco-2-enone

F) more

M

To a solution of /3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)ucyclohex-h-enone (1.0 equiv.) in degassed dioxane and 2M MagzhbOz was added 4-chloro -3-nitropyridine (1.2 equiv.) and Pa(arrOCl--DCM (0.05 equiv.). The reaction mixture was heated in an oil bath at 110 "C for 2 h. It was cooled to room temperature, then diluted with EOAc , added HgO - a dark solution with a lot of emulsified substance was formed. It was filtered to remove solids, then the organic phase was extracted, dried over Ma»5O: and concentrated. The crude substance was purified by chromatography on silica gel, eluting with ethyl acetate and hexanes (1:11) and 3-(3-nitropyridin-4-yl)ucyclohex-2-enone was obtained (55 90, 2 stages). LC/MS « 219 (MAN), LC :- 2.294 min.

Synthesis of 3-(3-nitropyridin-4-yl cyclohex-2-enol).

ON zi MO" more

M

To a solution of 3-(3-nitropyridin-4-yl)ucyclohex-2-enone (1.0 equiv.) was added EN (0.2 M) and CeSys-7HgO (1.3 equiv.). The reaction mixture was cooled to 0 °C, then MavVvVNa (1.3 equiv.) was added in portions.

Stirred for 2 hours. at 0 "C, then the reaction was stopped by adding water, concentrated to remove EUN, added E(OAc, extracted organic substances, washed with brine, then dried over Mag50" and concentrated to obtain 3-(3-nitropyridin-4-yl)ucyclohex - 2-enol (99 95). LC/MS - 221.1 (MN), LC - 2.235 min.

Synthesis of 2-(3-(3-nitropyridin-4-yl)ucyclohex-2-enyl)isoindoline-1,3-dione (F)

M c) with MO" more

M

To a solution of 3-(3-nitropyridin-4-yl)ucycloheco-2-enol (1.0 equiv.), triphenylphosphine (1.5 equiv.) and phthalimide (1.5 equiv.) in THF (0.3 M ) at 0"С, (E)-di-tert-butyldiazene-1,2-dicarboxylate (1.5 equiv.) was added dropwise. The reaction mixture was stirred at 0С for 2 h. It was concentrated to dryness under vacuum, then the crude substance was purified by column chromatography on silica gel eluting with EtOAc and hexanes (111 with the addition of 5 95 methanol) and 2-(3-(3-nitropyridin-4-yl)ucyclohex-2-enyl)isoindoline-1,3-dione was obtained (output 63 90) RH/MS - 350.3 (MN), RH - 3.936 min.

Synthesis of 2-(3-(3-aminopyridin-4-yl)ucyclohex-2-enyl)isoindoline-1,3-dione (F)

M c) more

M

Re (6.0 equiv.) and the reaction mixture was stirred at room temperature for 2 h.

It was filtered, then washed with methanol and the filtrate was concentrated. Ethyl acetate and a saturated solution of MaHsSOO were added to the crude substance, the organic substances were dried over Ma»boOx and concentrated to obtain 2-(3-(3-aminopyridin-4-yl)cyclohex-2-enyl)isoindolin-1,3-dione in the form thick yellow resin with a yield of 96 95. RH/MS - 320.0 (MAN), RH - 2.410 min.

Synthesis of 2-(3-(3Z-aminopyridin-4-yl)ucyclohexyl)isoindoline-1,3-dione (F)

M c) more

M

10 95 Pa/ C (0.2 equiv.) and the reaction mixture were stirred in an atmosphere of He supplied from a cylinder. Three days later, the reaction mixture was filtered through celite, washed with ethyl acetate and methanol, the filtrate was diluted with ethyl acetate and washed twice with a 2M solution of MgCO3. The organic phase was dried over magnesium sulfate, filtered and concentrated. The crude substance was triturated with hexanes and ethyl acetate and 2-(3-(3-aminopyridin-4-yl)ucyclohexyl)isoindolin-1,3-dione was obtained with a yield of 73 95. LC/MS - 322.2 (MH), RF- 0.64 min.

Synthesis of 5,5-dimethyl-3-oxocyclohex-1-enyltrifluoromethanesulfonate - - to

OT

In a 3-neck round-bottom flask, 5,5-dimethylcyclohexane-1,3-dione (1.0 equiv.) was dissolved in DCM (0.2 M). Sodium carbonate (1.1 equiv.) was added and the mixture was stirred under an atmosphere of M" while stirring with a magnetic stirrer in an ice bath/(the salt solution was cooled to - -5 "Sb. Trifluoromethanesulfonic acid anhydride (1.05 equiv.), diluted with DCM , was added dropwise through a dropping funnel for 90 min. After the addition was complete, the reaction mixture was stirred at - 0 "C for 1 h. According to LCMS and 1H NMR data, the starting substance still remained. Sodium carbonate (0.51 equiv.) was additionally added and trifluoromethanesulfonic anhydride (0.50 eq.) After 2 h, the mixture was filtered through a funnel with a coarse porous glass filter (the precipitate on the filter was washed with

DXM), transferred to an Erlenmeyer flask, the reaction mixture was neutralized to pH-7 by careful addition of a saturated aqueous solution of sodium bicarbonate with vigorous stirring, transferred to a separatory funnel and the layers were separated. The organic layer was washed with brine, dried over M95O", filtered and concentrated to obtain 5,5-dimethyl-3-oxocyclohex-1-enyltrifluoromethanesulfonate, which was used in the next stage without additional purification.

LC/MS (t/2): МНА-273.1, NI-1.03.

Synthesis of 5,5-dimethyl-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)ucyclohex-2-enone

My "V

I

All reagents: 5,5-dimethyl-3-oxocyclohex-1-enyltrifluoromethanesulfonate (1.0 equiv.), potassium acetate (3.0 equiv.), and bis(pinacolato)diboron (2.0 equiv.) were added to 1 ,4-dioxane (0.2 M) in a round-bottom flask and degassed by passing Me through the mixture for 10 min. The adduct Rasig(arry) - DXM (0.03 equiv.) was added and the reaction mixture was heated in an M" atmosphere overnight at 80 2С in an oil bath in a flask equipped with a reflux condenser. The mixture was cooled to room temperature, filtered through a funnel with a coarse porous glass filter, the precipitate on the filter was washed with 1,4-dioxane and 5,5-dimethyl-3-(4,4,5,5-tetramethyl-1,3, 2-dioxaborolan-2-yl)ucyclohex-2-enone in 1,4-dioxane, which was used in the next step without further purification. LC/MS (t/27): MH"" (boronic acid) -169.1, NI - 0.50.

Synthesis of 5,5-dimethyl-3-(3-nitropyridin-4-yl)ucycloheco-2-enone (c) d MO" more

M

Boronic acid ester of 5,5-dimethyl-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)cyclohex-2-enone (1.0 equiv.) was dissolved in 1, 4-dioxane in a round-bottom flask and degassed by passing Me2 through the solution for 30 min. 4-Chloro-3-nitropyridine (1.3 equiv.) and 2M aqueous sodium carbonate solution (2.0 equiv.) were added, and for 10 min. .

The reaction mixture was stirred at 110 "C for 2 h. The mixture was added to EOAc and water. The resulting mixture was filtered through Celite, the filter residue was washed with EOAc. The organic layer was separated and the aqueous layer was extracted with EOAc. The combined organic layers were washed with brine , dried over Ma5O», filtered and concentrated. The residue was purified by chromatography on silica gel (eluated with a mixture of EAs':hexanes - from 1:10 to 2:1) and 5,5-dimethyl-3-(3-nitropyridine- 4-Ilucycloheco-2-enone (46.7 95 for three stages). LC/MS (t/2): МН:-247 2,

VI-0.79.

Synthesis of 5,5-dimethyl-3-(3-nitropyridin-4-yl)ucycloheco-2-enol

ON d MO" sh

M

To a solution of 5,5-dimethyl-3-(3-nitropyridin-4-yl)ucyclohex-2-enone (1.0 equiv.) and CeCl2-7H2H2O (1.2 equiv.) in

Meon (0.2 M) was added at 0 °C with MavBnae (1.0 equiv). The solution was stirred for 1 h and then the reaction was stopped by adding 5 ml of water. The volatiles were removed in vacuo and the residue was partitioned between EAs and H2H0 The organic layer was separated and washed with brine.

The combined aqueous layer was back-extracted with EOAc and the organic matter was washed with brine. The combined organic substances were dried over Md5O", filtered and concentrated.

The residue was purified on a column (5 96 methanol in a 1:11 mixture of ethyl acetate and hexanes) and 5,5-dimethyl-3-(3-nitropyridin-4-yl)ucyclohexo-2-enol (74 95) was obtained. LC/MS (t/2): МНА-249.2, NI-0.76.

Synthesis of 2-(5,5-dimethyl-3-(3-nitropyridin-4-yl)-cyclohex-2-enyl)isoindoline-1,3-dione

(F)

M c) with MO" more

M

To a homogeneous solution of 5,5-dimethyl-3-(3-nitropyridin-4-yl)ucyclohexo-2-enol (1.0 equiv.) was added triphenylphosphine (1.5 equiv.) and phthalimide (1.5 equiv.) in THF (0.2 M), cooled to 0 "C, di-tert-butyl azodicarboxylate (1.5 equiv.) in THF was added to the solution. The mixture was stirred at 0 "C for 2 h.

The reaction mixture was concentrated in vacuo. The residue was purified on a column (5 95 methanol in a 1:1 mixture of ethyl acetate and hexanes) and 2-(5,5-dimethyl-3-(3-nitropyridin-4-yl)ucyclohex-2-enyl)isoindolin-1,3 -dione (99 95). LC/MS (t/2): MH--378.2, NI-1.10.

Synthesis of 2-(3-(33-aminopyridin-4-yl)-5,5-dimethylcyclohex-2-enyl)isoindoline-1,3-dione (F)

M c) with MN" more

M

A solution of 2-(5,5-dimethyl-3-(3-nitropyridin-4-yl)ucyclohex-2-enyl)isoindoline-1,3-dione (1 eq.) in acetic acid (0.1 M) was purged with nitrogen within 10 min. Then 10 95 Ra/C (0.10 equiv.) was added. The reaction mixture was stirred at room temperature overnight under a hydrogen atmosphere. The solids were removed by filtration through Celite, then washed with EtOAc and Meon. The filtrate was concentrated, diluted with EOAc and washed twice with a 2M aqueous solution of MaCoO3.

The organic layer was dried over Md5O5, filtered and concentrated. The residue was purified on a column (5 95 methanol in a 1:11 mixture of ethyl acetate and hexanes) and 2-(3-(3-aminopyridin-4-yl)-5,5-dimethylcyclohex-2-enyl)isoindolin-1,3- dion (89 95). LC/MS (t/2): MH--348.3, NI-O.79.

Synthesis of 2-(5-(3-aminopyridin-4-yl)-3,3-dimethylcyclohexyl)isoindoline-1,3-dione (F)

M c) d MN" what

M

A solution of 2-(3-(3-aminopyridin-4-yl)-5,5-dimethylcyclohex-2-enyl)isoindoline-1,3-dione (1.0 eq.) in acetic acid (0.1 M) was purged with nitrogen for 10 min. Then 10 95 Ra/C (0.1 equiv.) was added.

The reaction mixture was stirred at 45°C under 300 psi hydrogen in a steel autoclave overnight and at 65°C at 300 psi for 5 h. The solids were removed by filtration through Celite, then washed with EtOAc and Meon. The filtrate was concentrated, diluted with EOAc and washed twice with a 2M aqueous solution of MaCoO3.

The organic layer was dried over Md5O5, filtered and concentrated. The residue was purified on a column (5 95 methanol in a 1:11 mixture of ethyl acetate and hexanes) and 2-(5-(33-aminopyridin-4-yl)-3,3-dimethylcyclohexyl)isoindoline-1,3-dione was obtained (53 96 ). LC/MS (t/2): MH--350.3, NI-0.78. Enantiomerically pure 2-(18,5H8)-5-(3-aminopyridin-4-yl)-3,3-dimethylcyclohexyl)isoindoline-1,3-dione and 2-(15,55)-5-(3- aminopyridine -4-yl)-3,3-dimethylcyclohexyl)isoindoline-1,3-dione was isolated using chiral HPLC (for analysis K-7.526 min and 13.105 min, respectively; hexanes:ethanol-80:20 (vol.:0b6.) , SpigaIse! 0О-Н 100x4.6 mm at 1 ml/min. For preparative separation, hexanes:ethanol - 80:20 (vol.:vol.), SpigaIseI 0U-H (250x20 mm at 20 ml/min). " H NMR (SOCIi"): 5 8.04 (b, 1H), 8.00 (a, 1H), 7.82 (t, 2H), 7.71 (t, 2H), 7.06 (a, 1), 4.54 (t, 1H), 3.71 (t, 2H), 2.89 (t, 1H), 2.23-2.44 (t, 2H), 1.90 (t, 1H ), 1.20-1.60 (t, ZN), 1.18 (in, ZN), 1.07 (5, ZN).

Synthesis of 4-(cyclohexa-1,3-dienyl)-3-nitropyridine by MO»:

To a solution of 3-(3-nitropyridin-4-yl)cyclohex-2-enol (1.0 equiv.) was added dioxane (0.18 M) and p-TOC (p-toluenesulfonic acid) (1.1 equiv.) . The solution was heated at 100 "C for 4 hours. It was cooled to room temperature, treated with a saturated solution of ManHsSoOz and ethyl acetate, the organic phase was dried over Mag5O: and concentrated. The crude substance was purified by column chromatography on silica gel eluting with 100 96 DXM and obtained 4 -(cyclohexa-1,3-dienyl)-Z-nitropyridine in the form of a yellow oil (yield 27 95). LCMS (t/2): 203.1 (MNU), LC -3.53 min, "H-

NMR (SOSI»z): 9.02 (5, 1H), 8.70 (a, 9-5.3, 1H), 7.30 (a, 9-5.3, 1H), 6.15- 6.17 (t, 1H), 6.02-6.11 (t, 2H), 2.35-2.38 (t, 4H).

Synthesis of (w/-)-2-azido-4-(3-nitropyridin-4-yl)ucycloheco-3-enol

ON

Mi zi MO" more

M

To a solution of 4-(cyclohexa-1,3-dienyl)-3-nitropyridine (1.0 equiv.) in DCM (0.1 M) was added MansSoOz (1.2 equiv.) and a yellow solution was obtained. It was cooled to 0 "C, then MHPBC (1.0 equiv.) was added to the solution in one portion as a solid. The reaction mixture was stirred at 0 C for 3.5 h. The course of the reaction was monitored by TLC and LC/MS. The product is ionized in the form of MAN-237 (diol); KI-0.41 min in ORI C. The reaction was stopped with a saturated solution of Manso»z, then extracted with DCM (3 times). The organic phase was additionally washed with brine, then dried over Mag5O». :, was filtered and concentrated to give the crude epoxide as a yellow oil, which was used without further purification.

To a solution of the crude substance obtained above in EN and water (3:11) (turbid yellow solution) was added MaM»z (2.0 equiv.) and MNACI (2.0 equiv.) and a clear yellow-hot solution was obtained. The reaction mixture was stirred for 16 h, then concentrated. EIOAc and water were added, the organic phase was further dried over Md50O" and concentrated to give a brown oil. The oil was loaded onto silica gel and purified by column chromatography (IZCO, 0-50 90 EtOAc) to give (/-)-2-azido-4-(3-nitropyridin-4-yl)ucyclohexo-3-enol as a yellow oil (44 Fo in 2 stages). RHMS (t/2) - 262 (MNU), RH Ke2.35 min.

Synthesis of (17-)-4-(3-azido-4-(tert-butyldimethylsilyloxy)cyclohex-1-enyl)-3-nitropyridine

TV5

M hu MO more

M

To a solution of (17-)-2-azido-4-(3-nitropyridin-4-yl)ucycloheco-33-enol (1.0 equiv.) in DCM (0.15 M) at room temperature was added TVZSI (2, 0 equiv.), imidazole (2.0 equiv.) and DMAP (0.1 equiv.). After 18 hours water was added, the organic substances were washed with brine, then dried over Magn5Ox and concentrated.

The crude substance was loaded into silica gel and purified by column chromatography (LCC) eluting with ethyl acetate and hexanes (20 95). (/-)-4-(3-Azido-4-(tert-butyldimethylsilyloxy)cyclohex-1-enyl)-3-nitropyridine was obtained as a yellow oil with a yield of 60 95. LCMS (t/g): 376.3 ( MNU), RK K-5,848 min.

Synthesis of (1y-)-tert-butyl-6-(tert-butyldimethylsilyloxy)-3-(3-nitropyridin-4-yl cyclohex-2-enylcarbamate)

TV5

MNVoOs

With MO" more

M

(/-)-4-(3-azido-4-(tert-butyldimethylsilyloxy)cyclohex-1-enyl)-3-nitropyridine (1.0 equiv.) and pyridine (0.1 M) were added to a round-bottomed flask to obtain yellow solution. Ammonium hydroxide (10:1 pyridine:ammonium hydroxide) was added, followed by RMez (3.0 equiv). After 10 min, the reaction mixture turned dark brown. Stirred at room temperature for 1.5 hours. The reaction was stopped by adding EN and concentrated. Repeated 2 more times. A saturated solution of Manso»z and dioxane (1:1, 0.1M) was added to the crude substance. VosgO (1.0 equiv.) was added. Stirred for 1 hour. at room temperature. Washed with H2O and EIOAc, the organic phase was dried over

M95O5", filtered and concentrated. The residue was purified by column chromatography on silica gel (IZCO, 5:11 Nex/E(Ac). Pure fractions were collected and concentrated to give (h/-)-tert-butyl-6-(tert-butyldimethylsilyloxy)-3-(Z -nitropyridin-4-yl)ucyclohex-2-enylcarbamate in the form of a foamy substance.

Synthesis of (1Y-)-tert-butyl-3-(3-aminopyridin-4-yl)-6-(tert-butyldimethylsilyloxy)cyclohex-2-enylcarbamate tv5

MNVoOs from MN" more

M

To a solution of /((47-)-tert-butyl-6-(tert-butyldimethylsilyloxy)-3-(3-nitropyridin-4-yl cyclohex-2-enyl carbamate (1.0 equiv.) in ASON (0.18 M) Re (6.0 equiv.) was added and the reaction mixture was stirred for 20 h. The reaction mixture was worked up by diluting the reaction mixture with methanol, filtered and the filtrate was concentrated. To the crude material was added ethyl acetate and saturated Manso's solution, the organics were dried over sodium sulfate and concentrated and (/-)-tert-butyl-3-(3-aminopyridin-4-yl)-6-(tert-butyldimethylsilyloxy)cyclohex-2-enylcarbamate was obtained as a yellow oil with a yield of 94 95. LCMS (n/z): 420.3 (MNU), RH E-3.88 min.

Synthesis of (1Y-)-tert-butyl-5-(3-aminopyridin-4-yl)-2-(tert-butyldimethylsilyloxy)cyclohexylcarbamate tv5

МНВос х МН still

M

To a solution of ((47-)-tert-butyl-3-(3-aminopyridin-4-yl)-6-(tert-butyldimethylsilyloxy)cyclohex-2-enylcarbamate (1.0 equiv.) in MeOHN (0.1 M ) added Ra/cS (20 wt. 95) and the reaction mixture was stirred in an atmosphere of hydrogen supplied from a cylinder for 18 h. LC/MS of the reaction mixture indicated the presence of diastereoisomers, the reaction mixture was filtered, washed with E(OAc and the filtrate The crude substance was purified by preparative HPLC (dimethyl sulfoxide in DMSO) and the pure fractions were combined, neutralized with solid Manso»z, extracted with ethyl acetate, washed with brine, dried over Mag5O5 and concentrated to obtain product A (yield 8 9o) and product B ( exit 51 Ob).

Product A: RHMS (n/z): 422.4 (MNU), RH Ki-3.75 min.

Product B: RHMS (n/z): 422.4 (MNU), RH Ki-3.94 min.

Synthesis of 1,4-dioxaspiro|4,5|dec-7-en-v-yltrifluoromethanesulfonate gl ( and

OT

1,4-Dioxaspiro|4,5|decan-8-one (1.0 equiv.) was dissolved in ether (0.1 M) and stirred at -15 "С, then 1 M Manmose (1.05 equiv.) was added and was stirred for 70 min, then TO (1.05 eq.) was added and the reaction mixture was allowed to slowly warm to RT. The mixture was stirred for 28 h, washed with a saturated aqueous solution of ManNSO 3 and then with water. The aqueous layers were combined and extracted with ether.

The organic layers were combined, dried over Mo 5 O 5 , filtered and concentrated. The residue was purified on a column (ethyl ether:ohexanes - 1:4) and 1,4-dioxaspiroI4,5|dec-7-en-8-yltrifluoromethanesulfonate (65 95) was obtained. LC/MS (t/2): MH--289.0, Ne0.97. TOP Ki-3.77.

Synthesis of 4,4,5,5-tetramethyl-2-(1,4-dioxaspiro|4,5|dec-7-en-8-yl)-1,3,2-dioxaborolane background (

ВХ о о

A solution of 1,4-dioxaspiro|4,5|dec-7-en-8-yltrifluoromethanesulfonate (1.0 equiv) in dioxane (0.5 M) was purged with nitrogen for 30 min. Then 4,4,445,5,5,5'-octamethyl-2,2'-6i(1,3,2- dioxaborolane) (1.0 equiv.), COAc (3.0 equiv.), Ra(arr )CI.-DCM (0.2 equiv.) and the solution was stirred in a closed autoclave at 80 "C. The reaction mixture was filtered through a layer of celite, then ethyl acetate was added to the filtrate and washed with brine, dried over Mo950O54, filtered and concentrated.

The residue was purified on a column (ethyl acetate: hexanes - 1:1) and 4,4,5,5-tetramethyl-2-(1,4-dioxaspiro|4,5|dec-7-en-8-yl)-1 was obtained ,3,2-dioxaborolane (95 95). LC/MS (t/27): МНА-267.1, В-0.95.

Synthesis of 3-nitro-4-(1,4-dioxaspiro|(4,5|dec-7-en-8-yl)pyridine background 94.0 d MO»

M

DME solution (0.2 M) and 2 M aqueous sodium carbonate solution (1.7 equiv.) were purged with nitrogen for 20 min. Then 4-chloro-3-nitropyridine (1.6 equiv.), 4,4,5,5-tetramethyl-2-(1,4-dioxaspiro|4,5|dec-7-en-8-yl) was added -1,3,2-dioxaborolane (1.0 equiv.), Ra(arrOCI»--DCM (0.05 equiv.) and stirred in a closed autoclave at 110 "C. The reaction mixture was stirred at this temperature for 3.5 h. The reaction mixture was diluted with ethyl acetate, washed with water, dried over Ma5O»4, filtered and concentrated.

The residue was purified on a column (ethyl acetate: hexanes - 1:1 with the addition of 10 95 methanol) and 3-nitro-4-(1,4-dioxaspiro|4,5|dec-7-en-8-yl)pyridine (83 905). LC/MS (t/2): MH--263.2, NI-O.71.

Synthesis of 4-(3-nitropyridin-4-yl)ucycloheco-3-enone

F) d MO" sh

M

A mixture of 3-nitro-4-(1,4-dioxaspiro(|4,5|dec-7-en-8-yl)pyridine (1.0 equiv.) in 20 95 TFC in SneoSi» (0.2 M) stirred at room temperature overnight Solvents were removed under reduced pressure.

The residue was dissolved in ethyl acetate (200 ml) and washed with a saturated solution of ManHsSoOz (30 ml) and a saturated solution of Mas! (30 ml). The organic substances were dried over MozoO»4, filtered and concentrated to give 4-(3Z-nitropyridin-4-yl)ucycloheco-Z-enone (85 90). The crude product was used in the next step without additional purification. LC/MS (It/2): MH--218.9, B-0.60

Synthesis of 4-(Z-nitropyridin-4-yl)ucycloheco-3-enol on db MO» sh

M

Sodium borohydride (1.8 equiv.) was added to a solution of 4-(3-nitropyridin-4-yl)ucyclohex-3-enone (1.0 equiv.) in methanol (0.2 M) at 0 "C. The reaction mixture stirred at 0 "C for 2 hours. Methanol was removed under reduced pressure. The residue was dissolved in ethyl acetate (200 mL) and washed with saturated Massey solution (30 mL). The organic substances were dried over Mo950O5, filtered and concentrated to give 4-(3-nitropyridin-4-yl)ucyclohex-3-enol (85 95). The crude product was used in the next step without additional purification. LC/MS (t/7): MH--221.0, VI-0.55

Synthesis of 4-(3-nitropyridin-4-yl)ucyclohexo-3-enylmethanesulfonate

OM5 and MO"

M

To a solution of 4-(3-nitropyridin-4-yl)ucyclohexo-3-enol (1.0 equiv.) and DIPEA (diisopropylethylamine) (2.5 equiv.) in СНоСи» (0.15 M) at 0"С methanesulfonyl chloride (1.8 equiv) was added. The reaction mixture was stirred at 0 C for 1 h. The reaction mixture was diluted with ethyl acetate (200 mL) and washed with saturated Massey solution (30 mL). The organics were dried over Mo5O", filtered and concentrated to give 4-(3-nitropyridin-4-yl)cyclohex-3-enylmethanesulfonate (93 95).

The residue was used in the next stage without additional purification. LC/MS (t/2): MH--299.0,

VI-0.70

Synthesis of 4-(cyclohexa-1,3-dienyl)-3-nitropyridine d MO» sh

M

DBU (1,8-diazabicyclo/5.4.O1undec-7 -ene) (1.8 equiv.).

The reaction mixture was stirred at RT overnight. The reaction mixture was diluted with ethyl acetate (200 mL) and washed with saturated Massey solution (30 mL). The organic substances were dried over Mazoch, filtered and concentrated. The residue was purified on a column (5 95 methanol in a 1:1 mixture of ethyl acetate and hexanes) and 4-(cyclohexa-1,3-dienyl)-3-nitropyridine was obtained. LC/MS (t/2): МНА-203.2, NI-0.85.

Synthesis of (17-)-tert-butyl-6-hydroxy-3-(3-nitropyridin-4-yl)ucyclohexo-2-enylcarbamate

N

MNVos d MO" more

M

To a solution of (/-)-2-azido-4-(3-nitropyridin-4-yl)ucyclohex-3-enol (1.0 eq.) in pyridine and MNAOH (871, 0.23 M) at room temperature was added trimethylphosphine (3.0 equiv.). The mixture was stirred at room temperature for three hours. The solvents were removed. Ethanol was added to the residue. The ethanol was then removed in vacuo to ensure complete ammonia removal. The residue was dissolved in 1,4-dioxane and a saturated aqueous solution of sodium bicarbonate was added to the mixture, followed by BosgO (1.0 equiv.) in

THF. The resulting mixture was stirred at room temperature for 2 hours. The reaction mixture was diluted with ethyl acetate and washed with saturated Massey solution. Organic substances were dried over

Ma95O", filtered and concentrated. The residue was purified on a column (5 95 methanol in a 1:1 mixture of ethyl acetate and hexanes) and (/-)-tert-butyl-β6-hydroxy-3-(3-nitropyridin-4-yl cyclohex-2-enylcarbamate) was obtained (82 95) RH/MS (t/2): MH--336.0, VI-0.71

Synthesis of (1U-)-2-(tert-butoxycarbonylamino)-4-(3-nitropyridin-4-yl)ucyclohex-3-enylmethanesulfonate

M5

MNVos d MO" more

M

To a solution of (/-)-tert-butyl-6-hydroxy-3-(3-nitropyridin-4-yl cyclohex-2-enylcarbamate (1.0 equiv.) and triethylamine (1.5 equiv.) in СНоСи» (0 .2 M) at 0 "C, methanesulfonyl chloride (1.2 equiv.) was added. The mixture was stirred for 2 h at this temperature. The reaction mixture was diluted with ethyl acetate and washed with saturated Massey solution. The organic substances were dried over NaCl, filtered and concentrated to obtain (h/-)-2-(tert-butoxycarbonylamino)-4-(3-nitropyridin-4-yl cyclohex-3-enylmethanesulfonate (85 95), which was used in the next stage without additional purification.

LC/MS (t/2): МНА-414.0, Н-0.82

Synthesis of (7/-)-5-(Z-nitropyridin-4-yl)-3,Za,7,7a-tetrahydrobenzo|Ch|oxazol-2(6H)-one (c) 0-5

Ministry of Education and Culture

M

A mixture of (7-)-2-(tert-butoxycarbonylamino)-4-(3-nitropyridin-4-yl cyclohex-3-enylmethanesulfonate (1.0 eq.) and pyridine (0.21 M) was stirred in a microwave oven at 110 " C for 10 min. The pyridine was removed under reduced pressure. The residue was dissolved in ethyl acetate and washed with a saturated solution of the mass. The organics were dried over Md5O5", filtered and concentrated to give (-/-)-5-(3-nitropyridin-4-yl) -3,Za,7,7a-tetrahydrobenzo|Foxazol-2(6H)-one (8595), which was used in the next step without additional purification. LC/MS (t/27): МН-А-262,1, B -0.49.

Synthesis of (-/-)-tert-butyl-5-(Z-nitropyridin-4-yl)-2-oxo-Za, 6,7,7a-tetrahydrobenzo|Foxazole-Z(2H)-carboxylate (6) 0- 4

Me Vos d MO» schi

M

To a solution of (h7/-)-5-(3-nitropyridin-4-yl)-3,Za,7,7a-tetrahydrobenzo|H|oxazol-2(6H)-one (1.0 equiv.),

TEA (1.8 equiv.) and a catalytic amount of DMAP in SNecSig (0.19 M) were added at room temperature to di-tert-butyldicarbonate (1.2 equiv.). The reaction mixture was stirred for 1 hour. The reaction mixture was diluted with ethyl acetate (100 mL) and washed with saturated Massey solution (30 mL). Organic substances were dried over Md5O5, filtered and concentrated. The residue was purified on a column (5 o methanol in a 1:11 mixture of ethyl acetate and hexanes) and obtained (/-)-tert-butyl-5-(3-nitropyridin-4-yl)-2-oxo-Za, 6.7, 7a-tetrahydrobenzo|H|oxazole-Z(2H)-carboxylate (98 95). LC/MS (t/2): МН-А-306.0, Бе-0.75.

Synthesis of (1Y-)-tert-butyl-5-(3-aminopyridin-4-yl)-2-oxohexahydrobenzo|H|oxazole-3(2H)-carboxylate

0-4

M-Vos d MN" school

M

To a solution of (h7/-)-tert-butyl-5-(3-nitropyridin-4-yl)-2-oxo-Za, 6,7,7a-tetrahydrobenzo|H|oxazole-

C(2H)-carboxylate (1.0 equiv.) in methanol and ethyl acetate (1:11, 0.1 M) was added to 1095 Pa/C (0.1 equiv.).

The resulting mixture was stirred in an atmosphere of NO for two hours. The solid was removed by filtration. The filtrate was concentrated under reduced pressure and obtained (/-)-tert-butyl-5-(3-aminopyridin-4-yl)-2-oxohexahydrobenzo|H|oxazole-3(2H)-carboxylate (87 95), which was used for the next stage without additional purification. LC/MS (t/2): МН--334.1, Н-0.51.

Synthesis of 5-methyl-3-oxocyclohex-1-enyltrifluoromethanesulfonate - I

OT

To a solution of 5-methylcyclohexane-1,3-dione (1.0 equiv.) in DCM (0.5 M) was added MazbOz (1.1 equiv.) and cooled to 0 "С. For 1 h. at 0 "С in under a nitrogen atmosphere, TO (1.0 equiv.) in DCM (5.0 M) was added dropwise. After addition, the reaction mixture was stirred for 1 hour. at room temperature (dark purple solution). The solution was filtered and the filtrate was neutralized to pH-7 by careful addition of a saturated solution of MaHsSoOz with vigorous stirring. The solution was transferred to a separatory funnel and the layers were separated. The organic layer was washed with brine, dried over

Mag5O", was filtered, concentrated in vacuo and dried in high vacuum for 15 min to give 5-methyl-3-oxocycloheco-1-enyltrifluoromethanesulfonate as a clear yellow oil with a yield of 7895. The triflate decomposes on storage and must be used immediately in the next reactions РХ/МС-259.1/300.1 (MAN and MASNziSM); MKI-0.86 min, RH - 3.84 min. "H-NMR (400 MHz,

SOFI") 6 parts/million: 6.05 (5, 1H), 2.70 (da, 9-17.2, 4.3, 1H), 2.53 (aa, 9U-16.6, 3 ,7, 1H), 2.48-2.31 (t, 2H), 2.16 (ad, U-16.4, 11.7, 1H), 1.16 (a, 9-5.9, ZN).

Synthesis of 5-methyl-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)ucyclohex-2-enone - in to "In her

To a solution of 5-methyl-3-oxocyclohex-1-enyltrifluoromethanesulfonate (1.0 equiv.) in degassed dioxane (0.7 M) was added bis(pinacolato)diboron (2.0 equiv.), KoAc (3.0 equiv. ) and Ra(arr)CI»-DCM (0.03 equiv.).

The reaction mixture was heated at 80 "C for 10 hours (strong initial heating leads to the formation of yellow-hot foam in the upper part of the heated solution, the heating bath should be removed before the foam settles, reheated to 80 "C and at this temperature the foam does not is formed), then cooled to room temperature and filtered through a funnel with a coarse filter made of porous glass.

The sediment on the filter was additionally washed with dioxane and the filtrate was used in the next stage without additional purification. LC/MS (liquid chromatography/mass spectroscopy) - 155.1 (MAN of boronic acid); K-O, 41 min, RH - 1.37 min.

Synthesis of 5-methyl-3-(3-nitropyridin-4-yl)ucycloheco-2-enone

F)

With MO"

M

To a solution of 5-methyl-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)ucyclohex-2-enone (1.0 equiv.) in degassed dioxane (0.5 M) and 2M Ma»zhbOz (2 equiv.) was added 4-chloro-3-nitropyridine (1.3 equiv.) and

Ra(ary)Si»-DHM (0.05 equiv.). The reaction mixture was placed in a flask equipped with a reflux condenser and heated in an oil bath at 110 "С for 1 hour. It was cooled to room temperature, filtered through a layer of celite, the layer was washed with ethyl acetate, and the filtrate was concentrated in vacuo.

The residue was additionally evaporated at 80 "C in a rotary evaporator for 1 hour to remove intermediate products (MAN-101) containing boron by sublimation. The residue was partitioned between brine and ethyl acetate and the layers were separated, the aqueous phase was further extracted with ethyl acetate (4x) , the organic substances were combined, dried over sodium sulfate, filtered and concentrated.

The crude substance was purified by chromatography on silica gel in DCM and eluting with a mixture of 2-50 90 ethyl acetate and hexanes. The pure fractions were concentrated in vacuo and a yellow-hot oil was obtained. The oil was kept under high vacuum (7500 mtorr) with ignition crystals overnight and a yellow-hot solid was obtained. The solid was further purified by trituration with hexanes to give 5-methyl-3-(3-nitropyridin-4-yl)cyclohex-2-enone (48 95, 2 stages). LC/MS - 233.2 (MAN); EC -0.69 min, РХ - 2.70 min. "H-NMR (400 MHz, Sasi»z) 6 ppm: 9.31 (5, 1H), 8.88 (a, 9-51, 1H), 7.30 (a, U-5.1, 1H), 6.00 (a, 9-24, 1H), 2.62 (aa, 2-16.4, 3.5, 1H), 2.53-2.34 (t, ZN), 2.23 (aa, 9-16.1, 11.7, 1H), 1.16 (a, 9-6.3, ZN).

Synthesis of cis-(-/-)-5-methyl-3-(3-nitropyridin-4-yl)ucycloheco-2-enol and, AOoHn db MO» )

M

To a solution of 5-methyl-3-(3-nitropyridin-4-yl)ucycloheco-2-enone (1.0 equiv.) in EN (0.3 M) was added

Sesiz-7HgO (1.2 equiv.). The reaction mixture was cooled to 0 "C, then MavBna (1.2 equiv.) was added in portions. It was stirred for 1 h at 0 "C, then the reaction was stopped by adding water, concentrated to remove EN, EOAc was added, organic substances were extracted, washed brine, then dried over Mag250", filtered and concentrated to give cis-(-/-)-5-methyl-3-(3-nitropyridin-4-yl)ucyclohex-2-enol (94,905). РХ/МСО - 235.2 (M.N), РХ - 2.62 min.

Synthesis of 4-(3-(tert-butyldimethylsilyloxy)-5o-methylcyclohex-1-enyl)-3-nitropyridine 1, "o. t! 9 d MO" that

M

To a solution of 5-methyl-3-(3-nitropyridin-4-yl)ucyclohexo-2-enol (1.0 equiv.) in DMF (0.5 M) was added imidazole (4.0 equiv.) .5 equiv.). After stirring for 18 hours. the solution was partitioned between EtOAc and HO and separated. After additional washing with NgO (Zh) and

MassCl(za), drying over MozO», filtration and removal of solvents gave 4-(3-(tert-butyldimethylsilyloxy)-5-methylcyclohex-1-enyl)-3-nitropyridine (85 95). LC/MS - 349, 2 (MN), RH - 5.99 min.

Synthesis of 4-(3-(tert-butyldimethylsilyloxy)-5o-methylcyclohex-1-enyl)/pyridin-3-amine and, "ОХ р! 9 de МН" more

M

A heterogeneous solution of 4-(3-(tert-butyldimethylsilyloxy)-5-methylcyclohex-1-enyl)-3-nitropyridine (1.0 equiv.) and iron (6.0 equiv.) in acetic acid at a concentration equal to 0, 4 M, vigorously stirred for 2 hours. The mixture was then passed through a pad of Celite, eluting with Meon. After removal of volatile substances in a vacuum, the residue was dissolved in EIOAc, washed with MgCO3 (za), MaCl(zao), dried over MazO5, filtered and volatile substances were removed in a vacuum to obtain 4-(3-(tert-butyldimethylsilyloxy)-5-methylcyclohex -1-enyl)pyridin-3-amine (78,965). LCMS (t/2): 319.3 (MNU; LC 23.77 min.

Synthesis of 4-(3-(tert-butyldimethylsilyloxy)-o-methylcyclohexyl)pyridin-3-amine 1, code

And 9

S Ge dyO MN"

FI

M

10 95 palladium on carbon ( 0.1 equiv.). The obtained heterogeneous solution was placed in a hydrogen atmosphere and stirred for 15 hours. Then the mixture was filtered through a layer of celite eluting with methanol. Volatiles were removed in vacuo to give 4-(3-(tert-butyldimethylsilyloxy)-5-methylcyclohexyl)pyridine-3-amine, (9095). RHMS (t/2): 321.3 (MNU); RH KCH-3.85 min.

Synthesis of cis-(-/-)-benzyl-4-(3-(tert-butyldimethylsilyloxy)-o-methylcyclohexyl)pyridin-3-ylcarbamate pr

Wed

M

Benzyl-2,5-dioxopyrrolidine- 1-yl carbonate (1.1 equiv.) and DMAP (0.05 equiv.). After stirring for 16 h. benzyl-2,5-dioxopyrrolidin-1-yl carbonate (0.55 equiv.) and DMAP (0.03 equiv.) were also added during CT. After stirring for another 24 hours. benzyl-2,5-dioxopyrrolidin-1-yl carbonate (0.1 equiv.) and DMAP (0.03 equiv.) were also added during CT. After stirring for another 18 hours. the solution was partitioned between EIOAc and MgCO3(za) and separated.

After additional washing with MgSO3(za) (2x) and MacSivza), drying over M4azox", filtration and removal of solvents, cis-(-/-)-benzyl-4-(3-(tert-butyldimethylsilyloxy)-5-methylcyclohexyl was obtained )pyridin-3-ylcarbamate. The crude substance was used without treatment. RH/MS - 455.3 (MAN), RH :- 4.39 min.

Synthesis of cis-(-/-)benzyl-4-(3-hydroxy-o-methylcyclohexyl)pyridin-3-ylcarbamate pu

Surl city

M

A solution of cis-(-/-)-benzyl-4-(3-(tert-butyldimethylsilyloxy)-o-methylcyclohexyl)pyridin-3-ylcarbamate in a mixture of 1:2:1 6 n. NSI/THF/Meon at a concentration equal to 0.1 M was stirred at RT for 6 h. Then the pH value was adjusted to -/7/ by adding 6 n. Mahon and volatiles were removed in vacuo. The aqueous layer was extracted with EtOAc and the organic matter was washed with MacCl(za), dried over Ma5O5, filtered, and after removal of volatile substances in a vacuum, cis-(-/-)benzyl-4-(3-hydroxy-5o-methylcyclohexyl) was obtained pyridin-3-ylcarbamate. The crude substance was used without treatment. RH/MS - 341.2 (M.N), RH - 2.38 min.

Synthesis of cis-(-/-)-benzyl-4-(3Z-methyl-5-oxocyclohexyl)pyridin-3-ylcarbamate by db МНОБ2

FI

M

Dessa-Martin periodinan was added to a solution of cis-(-/-)-benzyl-4-(3-hydroxy-5-methylcyclohexyl)pyridin-3-ylcarbamate in wet СНеСи» at a concentration equal to 0.16 M at 0"С (1.5 eq.) and the solution was stirred for 18 h while it was heated to RT.

ESOAc 1:11 10 95 Ma»252Oz/MaHsSOOz(za» and separated. After additional washing with 1:1

95 Mag252Oz3/ManNsCoOzvza) (2x) and MaSi(za), drying over Md5O", filtration, removal of solvents and purification by chromatography on silica gel (75-100 96 E(OAc/hexanes) gave cis-(-n/-) - benzyl-4-(3-methyl-5-oxocyclohexyl)pyridin-3-ylcarbamate in the form of a white solid (53 95, 5 stages). LC/MS - 339.2 (M.H).

Synthesis of cis-(-/-)-benzyl-4-(-3-(benzylamino)-5-methylcyclohexyl)pyridin-3-ylcarbamate by МНОВ2

FI

M

A solution of // cis-(-/-)-benzyl-4-(3-methyl-5-oxocyclohexyl)pyridin-3-ylcarbamate (1.0 equiv.) and benzylamine (3.0 equiv.) in MeoN at a concentration, equal to 0.25 M, stirred at CT for 2 hours.

When cooling in a bath to -78 "С, I and VNa (1.1 equiv., 2.0 M in THF) were added, and the solution was allowed to heat up to RT during 16 h with stirring. The solution was subjected to distribution between ЕУАСс and МаНсоОзиз), separated, additionally washed with ManHsSoOz(za) and MaSi(za), dried over Mazoch, filtered, and after removal of volatile substances in a vacuum, cis-(-/-)-benzyl-4-(-3-(benzylamino)-5-methylcyclohexyl was obtained )pyridine-Z-ylcarbamate in the form of a 4:11 mixture of isomers with a preference for the all-cis isomer.LC/MS - 430.3 (MH), LC - 0.62 min.

Synthesis of cis-(-/-)-tert-butyl-(-3-(3-aminopyridin-4-yl)-o-methylcyclohexylcarbamate pu dyO MH»

FI

M

To a solution of cis-(-/-)-benzyl-4-(-3-(benzylamino)-5-methylcyclohexyl)pyridin-3-ylcarbamate (1.0 equiv.) in methanol at a concentration equal to 0.07 M was added 20 95 palladium hydroxide on charcoal (0.2 equiv.).

The obtained heterogeneous solution was placed in a hydrogen atmosphere and stirred for 14 hours. Then, the reaction mixture was purged with Ar, BosgO (1.0 equiv.) was added, and the solution was stirred for 8 h. More H2O (1.0 equiv.) was added and the solution was stirred for another 16 h. Then the mixture was filtered through a layer of celite eluting with methanol. After removal of volatile substances in vacuo, purification by chromatography on silica gel (2.5-2.5 MeEOH/CHNeCl with the addition of 0.1 95 DIEA) and recrystallization from 1095 EtOAc/hexanes gave cis-(-/-)-tert-butyl -(-3-(Z-aminopyridin-4-yl)-5-methylcyclohexylcarbamate (49 95). LCMS (t/g2): 306.3 (MNU), LC h-2.59 min. Pure enantiomers could be obtained using chiral chromatography.

Synthesis of (-/-)-4-(5-methylcyclohexa-1,3-dienyl)-3-nitropyridine

With MO"

M

To a solution of (h/-)-5-methyl-3-(3-nitropyridin-4-yl)ucyclohex-2-enol (1.0 equiv.) in dioxane (0.1 M) was added p-TSC (1.0 eq.), and the reaction mixture was stirred at 100 "C for 3 h. The solution was cooled to room temperature, then passed through a layer of neutral aluminum oxide while eluting with E(OAc) and obtained /(-/-)-4--5- methylcyclohexa-1,3-dienyl)-3-nitropyridine in the form of a yellow oil with a yield of 68 95. LC/MS - 217.1 (MAN), LC - 3.908 min.

Synthesis of (w/-)-2-azido-b-methyl-4-(3-nitropyridin-4-yl)ucycloheco-3-enol

ON

M zh MO" more

M

To a solution of (/-)-4-(5-methylcyclohexa-1,3-dienyl)-3-nitropyridine (1.0 eq.) in DCM (0.1 M) at 0 "C was added MHPBK (1.1 eq. .) and the reaction mixture was allowed to warm to room temperature. After 3 h, the reaction was quenched with saturated Manso solution", extracted with DCM, and the organic phase was dried over sodium sulfate, filtered and concentrated to give a yellow oil. The crude substance was dissolved in ethanol and water ( 3:1, 0.1 M) and sodium azide (2.0 equiv.) and ammonium chloride (2.0 equiv.) were added. The reaction mixture was stirred for 4 h, then concentrated in vacuo. To the crude material was added ethyl acetate and water, the organic phase was washed with brine, dried over sodium sulfate, filtered and concentrated. 4-(Z-nitropyridin-4-yloxycycloheco-3-enol) in the form of an oil with a yield of 49 95. LC/MS - 276.1 (MH), 0.71 min.

Synthesis of tert-butyl-(-/-)-6-hydroxy-5-methyl-3-(3-nitropyridin-4-yl)ucyclohex-2-enylcarbamate

ON oMNVos zh MO" more

M

To a solution of (17-)-2-azido-6-methyl-4-(3-nitropyridin-4-yl)ucyclohexo-3-enol (1.0 equiv.) in pyridine and ammonium hydroxide (8:1, 0, 08 M) was added trimethylphosphine (3.0 equiv.) and the brown solution was stirred at room temperature for 2 h. Ethanol was added to the mixture and the solution was concentrated in vacuo (2x). Then the crude mixture was dissolved in dioxane and a saturated solution of ManNsoz (171, 0.08 M) and VosgO (1.0 equiv.) was added. The solution was stirred at room temperature for 2 hours, then partitioned between ethyl acetate and water. The organic phase was dried over magnesium sulfate, filtered and concentrated in vacuo. The crude product was purified by column chromatography on silica gel eluting with ethyl acetate and hexanes (1:1) to give tert-butyl-(i/-)-6-hydroxy-5-methyl-3-(3-nitropyridin-4-yl cyclohex- 2-enylcarbamate with a yield of 6995. LC/MS - 350.1 (MAN), -0.76 min.

Synthesis of (1Y-)-2-(tert-butoxycarbonylamino)-6-methyl-4-(3-nitropyridin-4-yl)ucyclohex-3-enylmethanesulfonate (FI "MNVos zu MO"

M

To a solution of tert-butyl-(-/-)-6-hydroxy-5-methyl-3-(3-nitropyridin-4-yl)ucyclohex-2-enylcarbamate (1.0 equiv.) in DCM (0.09 M ) triethylamine (1.5 equiv.) was added. The reaction mixture was cooled to 0 "C and M5ClII (1.2 equiv.) was added to the reaction mixture and stirred for 3 h. Water was added to the solution, the organic phase was dried over sodium sulfate, filtered and concentrated. The crude substance was purified by column chromatography on silica gel eluted with ethyl acetate and hexanes (111) and obtained /(h/-)-2-(tert-butoxycarbonylamino)-6-methyl-4-(3-nitropyridin-4-yl)cyclohex-3-enylmethanesulfonate in the form of a white foam substances with a yield of 65 95. LC/MS - 428.2 (MN), Ch-0.88 min.

Synthesis of (1Y-)-tert-butyl-7-methyl-5-(3-nitropyridin-4-yl)-2-oxo-Za,b,7,7a-tetrahydrobenzo|N|oxazole-Z(2H)-carboxylate at 0-5

M MVos with MO"

M

A solution of (1N-)-2-(tert-butoxycarbonylamino)-6-methyl-4-(3-nitropyridin-4-yl)ucyclohex-3-enylmethanesulfonate (1.0 eq.) in pyridine (0.2 M) in microwave oven dishes were heated at 110 C for 10 min. The yellow-hot solution was then concentrated to dryness and worked up by partitioning between ethyl acetate and water. The organic phase was dried over sodium sulfate, filtered and concentrated. The crude substance was dissolved in DCM (0.2 M) and triethylamine (1.8 equiv.), then VosgO (1.2 equiv.) was added to the reaction mixture. After stirring at room temperature for 40 min, the reaction mixture was concentrated in vacuo and purified by column chromatography on silica gel eluting with ethyl acetate and hexanes (1:1) to give (/-)-tert-butyl-7-methyl-5-(Z -nitropyridin-4-yl)-2-oxo-Za, 6,7,7a-tetrahydrobenzo|Ch|oxazole-Z(2H)-carboxylate in the form of a white foamy substance with a yield of 66 95. LC/MS - 376.0 ( MAN), CI-0.87 min.

Synthesis of (1Y-)-tert-butyl-5-(3-aminopyridin-4-yl)-7-methyl-2-oxohexahydrobenzo|hCh|oxazole-3(2H)-carboxylate (c) 0-X x MVos with MH »

M

To the solution of (1U-)-tert-butyl-7-methyl-5-(3-nitropyridin-4-yl)-2-oxo-Za, 6,7,7a-tetrahydrobenzo|H|Ioxazole-Z(2H)- carboxylate (1.0 equiv.) in Meon and ethyl acetate (11, 0.07 M) was added 95 Pa/C (0.1 equiv.) and the reaction mixture was stirred at room temperature under a hydrogen atmosphere.

After completion of the reaction, the solution was filtered through a layer of Celite, washed with Meon and ethyl acetate, the filtrate was concentrated to dryness in vacuo to obtain (h/-)-tert-butyl-5-(3-aminopyridin-4-yl)-7-methyl-2 -oxohexahydrobenzo|Ch|oxazole-Z3(2H)-carboxylate in the form of a mixture of diastereoisomers with a yield of 299 95. LC/MS - 348.2 (MAN), K-0.50 min.

Synthesis of (17-)-6-bromo-5-methyl-3-(3-nitropyridin-4-yl)ucyclohex-2-enol

Hg

AN xx MO" this

M

To a solution of 4-(5-methylcyclohexa-1,3-dienyl)-3-nitropyridine (1.0 equiv.) in THF and water (1:1, 0.13 M) was added MV5 (1.5 equiv.) and the reaction mixture was stirred at room temperature for 30 min.

After completion of the reaction, ethyl acetate and water were added to the reaction mixture, the organic phase was washed with brine, then dried over sodium sulfate, filtered and concentrated. The crude substance was purified by column chromatography on silica gel eluting with ethyl acetate and hexanes (1:1) to give (/-)-6-bromo-5-methyl-3-(3-nitropyridin-4-yl)ucyclohex-2-enol in the form of a yellow oil with a yield of 80 95. LC/MS - 315.0/313.0 (MAN), LC - 2.966 min.

Synthesis of (h/-)-2-azido-b-methyl-4-(3-nitropyridin-4-yl)ucycloheco-3-enolone: M zy MO» more

M

Potassium tert-butoxide was added to a solution of (h7/-)-6-bromo-5-methyl-3-(3-nitropyridin-4-yl cyclohexo-2-enol (1.0 eq.) in THF (0.1 M) (1.5 eq.). The reaction mixture turned from yellow-brown to black almost immediately. According to TLC, product formation was complete after 30 min. The reaction was stopped by adding a saturated solution of ammonium chloride and ethyl acetate. The organic phase was washed with brine, then dried over sodium sulfate, filtered and concentrated. The crude product was dissolved in ethanol and water (3:11, 0.1 M) and ammonium chloride (2.0 equiv.) and sodium azide (2.0 equiv.) were added. The dark yellow-hot reaction mixture was stirred at room temperature temperature during the night

LC/MS conversion to product was being completed. The reaction mixture was concentrated to remove ethanol, ethyl acetate and water were added and the organic phase was dried over sodium sulfate, filtered and concentrated. The crude substance was purified by column chromatography on silica gel eluting with ethyl acetate and hexanes (1:1) to give (h/-)-2-azido-b-methyl-4-(3-nitropyridin-4-yl)ucycloheco-3- enol with a yield of 55 95. LC/MS - 276.0 (MA-H), LC - 2.803 min.

Synthesis of (17-)-tert-butyl-6-hydroxy-5-methyl-3-(3-nitropyridin-4-yl)ucyclohex-2-enylcarbamate "МНВОос жи МО" more

M

To a solution of (17-)-2-azido-6-methyl-4-(3-nitropyridin-4-yl)ucyclohexo-3-enol (1.0 equiv.) in pyridine and ammonium hydroxide (8:1, 0, 08 M) was added trimethylphosphine (3.0 equiv.) and the brown solution was stirred at room temperature for 2 h. After completion of the reaction, EN was added and the solution was concentrated in vacuo. Ethanol was added and the reaction mixture was re-concentrated. Dioxane and a saturated solution of ManHsSoOz (1:1, 0.08 M), then BosgO (1.0 equiv.) were added to the crude substance. The reaction mixture was stirred at room temperature for 2 hours, then water and ethyl acetate were added. The organic phase was dried over Md5oO5x and concentrated. The crude product was purified by column chromatography on silica gel eluting with ethyl acetate and hexanes (1-1) and obtained (1U-)-tert-butyl-b-hydroxy-5-methyl-3-(3-nitropyridin-4-yl)ucyclohex -2-enyl carbamate (59 95). LC/MS - 350.1 (MAN), KE 0.76 min.

Synthesis of (17-)-2-(tert-butoxycarbonylamino)-b-methyl-4-(3-nitropyridin-4-yl)ucyclohexo-3-enyl acetate

OAS and MNVOos

Zh MO" more

M

To a solution of (/-)-tert-butyl-6-hydroxy-5-methyl-3-(3-nitropyridin-4-yl)ucyclohex-2-enylcarbamate (1.0 equiv.) in pyridine (0.1 M) AsgO (2.0 equiv.) was added and the reaction mixture was stirred at room temperature overnight. After completion of the reaction, the reaction mixture was concentrated to dryness, then treated with ethyl acetate and water. The organic phase was washed with brine, then dried over sodium sulfate, filtered and concentrated to give (/-)-2-(tert-butoxycarbonylamino)-6b-methyl-4-(3-nitropyridin-4-yl)ucyclohecon-3-enyl acetate with output 94 95. LC/MS - 392.2 (MN), Ch-0.94 min.

Synthesis of (17-)-4-(3-aminopyridin-4-yl)-2-(tert-butoxycarbonylamino)-b-methylcyclohexyl acetate

ОАс - МНВос х МН» more

M

To a degassed solution of (1N-)-2-"tert-butoxycarbonylamino)-6-methyl-4-(3-nitropyridin-4-yl)ucyclohex-3-enyl acetate (1.0 equiv.) in MeOH and EAs (1: 1, 0.1 M) was added to 10 95 Ra/С (0.1 equiv.) and the reaction mixture was stirred for 3 days at room temperature in an atmosphere of hydrogen supplied from a cylinder. After completion of the reaction, the solution was filtered through a layer of celite, the layer was washed with ethyl acetate, and the filtrate was concentrated. The crude material contained approximately 10 95 of the undesired isomer. The crude substance was dissolved in ethyl acetate (20 95) and hexanes and heated until all substances dissolved. The solution was kept at room temperature for 2 days. Then the precipitate was collected and (/-)-4-(3-aminopyridin-4-yl)-2-(tert-butoxycarbonylamino)-6-methylcyclohexyl acetate was obtained as a pure product with a yield of 59 95. LC/MS - 364.3 ( MN), CI-0.63 min.

Synthesis of 2-(tert-butoxycarbonylamino)-6-methyl-4-(3-nitropyridin-4-yl)ucyclohex-3-enylmethanesulfonate

OM5 - the same MNVOs or MO" more

M

Triethylamine (1, 5 equiv.) and the reaction mixture was cooled to 0 °C. M5CI (1.2 equiv.) was added to the reaction mixture and stirred for 3 h. Another 1.0 equiv. M5CI was added to the reaction mixture and stirred for another 2 h. the mixture was treated by adding water, the organic phase was washed with brine, dried over sodium sulfate and concentrated.

The crude product was purified by silica gel column chromatography eluting with ethyl acetate and hexanes (1:1) to give 2-(tert-butoxycarbonylamino)-6-methyl-4-(3-nitropyridin-4-ylylcyclohex-3-enylmethanesulfonate) as a white foamy substance with a yield of 65 95. LC/MS - 428.2 (MN), LC: 3.542 min.

Synthesis of (1Y-)-tert-butyl-7-methyl-5-(3-nitropyridin-4-yl)-2-oxo-Za,b,7,7a-tetrahydrobenzo|N|oxazole-Z(2H)-carboxylate (in; 0-5

MVos in the Ministry of Defense" more

M

A solution of (1U-)-2-(tert-butoxycarbonylamino)-6-methyl-4-(3-nitropyridin-4-yl)ucyclohex-3-enylmethanesulfonate (1.0 equiv.) in pyridine (0.2 M) was heated in a microwave oven at 110 "C for a minute. Then the yellow-hot reaction mixture was concentrated in vacuo, the crude substance was dissolved in ethyl acetate and water, the organic phase was dried over sodium sulfate and concentrated in vacuo.

The crude substance was dissolved in DCM (0.2 M), triethylamine (1.8 equiv.) was added, followed by VosgO (1.2 equiv.). The reaction mixture was stirred for 40 min, then concentrated to dryness. The crude substance was purified by column chromatography on silica gel eluting with a mixture of hexane and ethyl acetate (1:71) to give (/-)-tert-butyl-7-methyl-5-(3-nitropyridin-4-yl)-2-oxo -By,

6,7,7a-tetrahydrobenzo|chCh|oxazole-Z(2H)-carboxylate in the form of a white foamy substance with a yield of 66 95.

RH/MS - 376.0 (MAN), MOVEMENT: 3.424 min.

Synthesis of (1Y-)-tert-butyl-5-(3-aminopyridin-4-yl)-7-methyl-2-oxohexahydrobenzoic acid oxazole-3(2H)-carboxylate 0-4

MVos still

M

To the degassed solution /(17-)-tert-butyl-7-methyl-5-(3-nitropyridin-4-yl)-2-oxo-Za, 6,7,7a-tetrahydrobenzo|H|Ioxazole-Z(2H )-carboxylate (1.0 equiv.) in MeOH and EtOAc (1:1, 0.1 M) was added 10 95

Ra/C (0.1 equiv.). The reaction mixture was stirred overnight in an atmosphere of hydrogen supplied from a cylinder. After completion of the reaction, the solution was filtered through a layer of celite and the layer was washed with ethyl acetate. The filtrate was concentrated in vacuo and (/)-tert-butyl-5-(3-aminopyridin-4-yl)y-7-methyl-2-oxohexahydrobenzo|N|oxazole-3(2H)-carboxylate was obtained as the desired product in the form yellow foamy substance with a yield of 93 95. LC/MS - 348.1 (MAN), ECH-055 min.

Synthesis of ((147-)-(18, 28, 65)-6-methyl-4-(3-nitropyridin-4-yl)ucyclohexo-3-ene-1,2-diol and (w/-)-(18 , 25, 65)-6-methyl-4-(3-nitropyridin-4-yl)ucyclohex-3-en-1 2-diol)

ON ON tp, «on t, Finn! the differences between the Ministry of Defense and the Ministry of Defense"

An.) | AND.)

M M

To a solution of (/-)-(15, 55, 65)-6-bromo-5-methyl-3-(3-nitropyridin-4-yl)cyclohex-2-enol (1.0 equiv.) in

Potassium tert-butoxide (1.5 equiv.) was added to THF (0.1M) at room temperature. The reaction mixture was stirred for 10 min. The reaction was stopped with MNACI solution and treated with EOAc and washed with water and brine. The organic layer was dried over anhydrous sodium sulfate, filtered and dried under vacuum. The crude product was used in the next step without additional purification. KCh-0.5 (50 95 EOAc/hexanes). LCMS: MH.251.2 (in the form of diol), Ech-0.49 min. To a solution of crude (/-)-4-((15, 55)-o-methyl-7-oxabicyclo|4.1.0|hept-2-en-3-yl)-3-nitropyridine (1.0 equiv.) acetic acid (0.3 equiv.) was added to a mixture of 2:11 SNISM/HgO (0.1 M) at room temperature.

The reaction mixture was stirred for 16 h. at room temperature. Then the reaction was stopped with Mansoz solution, the reaction mixture was concentrated to remove most of the CH3ZSM, and the residue was partitioned between EIOAc and water. The combined organic layer was washed with water and brine, dried over anhydrous sodium sulfate, filtered and concentrated in vacuo. A mixture of diols ((/- )-1β8, 28, 65)-6-methyl-4-(3-nitropyridin-4-yl cyclohexo-3-ene-1,2-diol and (/-)-(18, 25, 65)-6-methyl-4-(3-nitropyridin-4-yl)ucyclohexo-3-ene-1,2-diol) was obtained by column flash chromatography as a white solid with a yield of 33.1 95. No -0.3 (100 95 EIHUAc; diols were not separated by TLC). RHMS: MNYA251.2, Ri-0.49 min.

Synthesis of (z/-)-4-((35, 48, 55)-3,4-bis(tert-butyldimethylsilyloxy)-5-methylcyclohex-1-enyl)-3-nitropyridine and (z/-)-4- ((3А8, 48, 55)-3,4-bis(tert-butyldimethylsilyloxy)-5-methylcyclohex-1-enyl)-3-nitropyridine

OotTvomM5 OotTvomM5 p, OotTvomM5 /p,, "OotTvom5 yin shchi iy i

M

(n/-) (n/-)

To a solution of a mixture of diols (1.0 equiv.) in DMF (0.3 M) at room temperature was added TVOMZSIII (7.0 equiv.) and imidazole (9 equiv.). The reaction mixture was stirred at room temperature overnight.

Then the reaction was stopped with saturated Mansoz solution. The reaction mixture was extracted using

EUAs. The combined organic layer was washed with water and brine, dried over anhydrous sodium sulfate, filtered and concentrated in vacuo. The mixture was successively purified by automatic column chromatography on silica (gradient mode with elution with EOAc and hexanes) and preparative reversed-phase HPLC ( 55 95-95 95 acetonitrile in water, then 5995-9595 acetonitrile in water) and obtained (/-)-4-((35, 48, 55)-3,4-bis(tert-butyldimethylsilyloxy)-5-methylcyclohex- 1-enyl)-3-nitropyridine (27,2 95) and (w/-)-4-((3А8, 48, 55)-3,4-bis(tert-butyldimethylsilyloxy)-5-methylcyclohex-1-enyl )-Z-nitropyridine (50.2 965). RHMS: МНя479.2,

Vi-1.60 and 1.63 min.

Synthesis of 4-(15, 35, 45, 5H8)-3,4-bis(tert-butyldimethylsilyloxy)-5-methylcyclohexyl)pyridin-3-amine and 4-18, ZB, 48, 55)-3,4-bis (tert-butyldimethylsilyloxy)-5-methylcyclohexyl)pyridin-3-amine atom OotvoMme oTvoMe no ptustme no, - tah ootszhtya also -- -i as chiral M chiral

To a solution of (-/-)-4-((3A8, 48, 55)-3,4-bis(tert-butyldimethylsilyloxy)-5-methylcyclohex-1-enyl)-3-nitropyridine (1.0 equiv.) in 10 95 palladium on charcoal (0.1 equiv.) was added to the ethanol/EOAc mixture at a concentration equal to 0.1 M. The obtained heterogeneous solution was placed in a hydrogen atmosphere and stirred for 14 hours. Then the mixture was filtered through a layer of celite eluting with EtOAc.

Volatiles were removed in vacuo and the crude was purified by automatic column chromatography on silica (K-0.2, 40,956 EOAc in heptane) to give the pure racemic product. RHMOS: МН-я451.3, КИ-1.35 min. The racemic compound was separated by chiral chromatography (IC column, 1 mL/min, 5 95 IPS (isopropyl alcohol) in heptane) to give 4-(15, 35, 45, 58)-3,4-bis(tert-butyldimethylsilyloxy) -5-methylcyclohexyl)pyridin-3-amine (6.01 min) and 4-(18, ZA, 48, 55)-3,4-bis(tert-butyldimethylsilyloxy)-5-methylcyclohexyl)pyridin-3-amine ( 8.34 min).

Synthesis of 4-(18, ZA, 45, 5H8)-3,4-bis(tert-butyldimethylsilyloxy)-5-methylcyclohexyl)pyridin-3-amine and 4-((15, 35, 4H, 55)-3,4 -bis(tert-butyldimethylsilyloxy)-5-methylcyclohexyl)pyridin-3-amine

OZ tesmMO OotvOomMo pes no How. Me's motto

De Tl, MN; not in

M zralny M chiral

To a solution of (-/-)-4-((35, 48, 55)-3,4-bis(tert-butyldimethylsilyloxy)-5-methylcyclohex-1-enyl)-3-nitropyridine (1.0 equiv.) in 10 95 palladium on carbon (0.1 equiv.) was added to ethanol at a concentration equal to 0.1 M. The obtained heterogeneous solution was placed in a hydrogen atmosphere and stirred for 14 hours.

Then the mixture was filtered through a layer of celite eluting with ethanol. Volatiles were removed in vacuo and the crude was purified by automatic column chromatography on silica (K-0.2, 40 906 EUAs in heptane) to give the pure racemic product (50.4 95).

RHMS: MNya451.3, E-1.35 min. The racemic compound was separated by chiral chromatography (IC column, 1 mL/min, 595 IPS in heptane) to give 4-(18, ZA, 45, 5H)-3,4-bis(tert-butyldimethylsilyloxy)-5-methylcyclohexyl) Pyridin-3-amine (6.98 min) and 4-(15, 35, 48, 55)-3,4-bis(tert-butyldimethylsilyloxy)-5-methylcyclohexyl)pyridin-3-amine (8.67 min) .

Synthesis of sodium 6-(methoxycarbonyl)-3-oxo-5-(trifluoromethyl)cyclohex-1-enolate

Svv 07 Va; (c) OM

Ethyl acetoacetate (1.0 equiv.) was added dropwise to a freshly prepared solution of sodium (1.0 equiv.) in I1-VChON (1 M) and the mixture was stirred in an ice bath for another 15 min. Ethyl 4,4,4-trifluorocrotonate (1.0 equiv.) was added dropwise and the mixture was stirred at room temperature for another 30 min. After boiling under reflux for 2 hours. the mixture was cooled and hexanes were added. The precipitate was filtered and sodium 6-(methoxycarbonyl)-3-oxo-5-(trifluoromethyl)cyclohex-1-enolate (4695) was obtained without further purification. LC/MS (t/27): MH: - 253.1, EC-0.70 min.

Synthesis of 5-(trifluoromethyl)cyclohexane-1,3-dione sodium 6-(Methoxycarbonyl)-3-oxo-5-(trifluoromethyl)cyclohex-1-enolate (1.0 equiv.) was dissolved in 1M

Mahon (1.0 eq.) and the mixture was refluxed for 1 h. After cooling to room temperature, the mixture was acidified with 5 M sulfuric acid. The mixture was extracted with ESUACS. After washing with water, the organic layer was dried over magnesium sulfate, the solvent was removed under reduced pressure and 5-(trifluoromethyl)cyclohexane-1,3-dione was obtained, which was used in the next stage without additional purification (98 95). РХ/MS (t/27): МН" -1811,

E-0.55 min.

Synthesis of 3-oxo-5-(trifluoromethyl)cyclohex-1-enyltrifluoromethanesulfonate

OT

To a suspension of 5-trifluoromethyl)cyclohexane-1,3-dione (1.0 equiv.) in DCM (0.23 M) was added TEA (1.2 equiv.) and a clear solution was obtained. The mixture was cooled to 0 °C. Then 20 (1.05 eq.) in DCM was added dropwise. The reaction mixture was stirred at this temperature for 2 h. The reaction mixture was diluted with DCM and washed with water, aqueous Mansoz solution, brine and dried over

MaBO:, was filtered and concentrated to obtain 3-oxo-5-(trifluoromethyl)cyclohex-1-enyltrifluoromethanesulfonate, which was immediately used in the next stage. LC/MS (t/g2): MH" - 313.0, 1.02 min.

Synthesis of 3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-5-(trifluoromethyl)cyclohex-2-enone

EzbU Zh "V o o

All reagents: 3-oxo-5-(trifluoromethyl)cyclohex-1-enyltrifluoromethanesulfonate (1.0 equiv.), Massas (3.0 equiv.), and bis(pinacolato)diboron (2.0 equiv.) were added to 1, of 4-dioxane (0.23 M) in a round-bottom flask and degassed by passing Me" through the mixture for 10 min. Rasiharri adduct).SNesi" (0.1 equiv.) was added and the reaction mixture was heated in an M2 atmosphere for 2 h. at 80 "C in an oil bath in a flask equipped with a reflux condenser. The mixture was cooled to room temperature, filtered through a funnel with a coarse porous glass filter, the precipitate on the filter was washed with "10 ml of 1,4-dioxane and obtained 3-(4 ,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-5-(trifluoromethyl)cyclohex-2-enone in 1,4-dioxane, which was immediately used in the next stage. LC/MS ( t/27): MH - 209.1 (boronic acid), CI - 0.60 min.

Synthesis of 3-(3-nitropyridin-4-yl)-5-(trifluoromethyl)cyclohex-2-enone

Es F) d MO" ;

M

Boronic acid ester 3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-5-(trifluoromethyl)cyclohex-2-enone (1.0 equiv.) was dissolved in 1, 4-dioxane (0.14 M) in a round-bottom flask was degassed by passing MeHg through the solution for 30 min. 4-Chloro-3-nitropyridine (1.3 eq.) and an aqueous solution of MgCO3 (2M, 2.0 eq.) were added, and Me was passed for 10 min, and then the adduct was added

Rasikh(arry).SNesSi (0.1 equiv.). The reaction mixture was stirred at 100 "C for 2 h. EAs and brine were added to the mixture. The resulting mixture was filtered through celite, the filter residue was washed with EtOAc. The organic layer was separated and washed with brine, dried over Mo5ox, filtered and concentrated. The crude product was purified by chromatography on silica gel (eluated with a mixture of E(OAc':hexanes - from 1:10 to 2:1) and obtained 3-(3-nitropyridin-4-yl)-5-(trifluoromethyl)cyclohex-2-enone (73 95 from the diketone in three stages.) LC/MS (t/2): MH - 287.1, Ch-0.85 min.

Synthesis of cis-3-(3-nitropyridin-4-yl)-5-(trifluoromethyl)cyclohex-2-enol

EzS,, hoNn zu MO"

Mo 3-(3-Nitropyridin-4-yl)-5-(trifluoromethyl)cyclohexo-2-enone (1.0 eq.) was mixed with cerium(II) chloride heptahydrate (1.0 eq.) and absolute ethanol ( 0.17). The mixture was stirred at ambient temperature until all solids dissolved. The mixture was cooled in an ice bath, and Mavn" (1.2 equiv.) was added in portions. The reaction mixture was stirred in an ice bath for 1 hour. The mixture was diluted with EtOAc, washed with water, dried over NaOH, filtered and concentrated. The residue was purified on a column (1:11 ethyl acetate and hexanes) to give cis-3-(3-nitropyridin-4-yl)-5-(trifluoromethyl)cyclohexo-2-enol (66 96). LC/MS (t/2): MH - 289.2, -0.72 min.

Synthesis of cis-4-(3-azido-5-(trifluoromethyl)cyclohex-1-enyl)-3-nitropyridine

EzS,,, MMZ with MO"

Spit

To a solution of cis-3-(3-nitropyridin-4-yl)-5-trifluoromethyl)cyclohex-2-enol (1.0 equiv.) in DCM (0.14

M) at room temperature, TEA (2.5 equiv.) and then M5Cl (1.8 equiv.) were added. The reaction mixture was stirred at room temperature for 2 hours. The solvent was removed. The residue was dissolved in

DMF (0.19 M) and then sodium azide (1.2 equiv.) was added to the mixture. The resulting mixture was stirred at room temperature for 1 hour. Another 1.2 equiv was added. sodium azide. The mixture was stirred at room temperature overnight. The reaction mixture was diluted with ethyl acetate and heptane and washed with saturated Massey solution. Organic substances were dried over M95O", filtered and concentrated. The residue was purified on a column (1:1 ethyl acetate and hexanes) to give cis-4-(3-azido-5-(trifluoromethyl)cyclohex-1-enyl)-3-nitropyridine (58 95). LC/MS (t/2): MH - 314.1, Ch-0.96 min.

Synthesis of tert-butyl-(18, ZA, 55)-3-(3Z-aminopyridin-4-yl)-5-(trifluoromethyl)cyclohexylcarbamate background with МН»

WITH

M

A solution of cis-4-(3-azido-5-(trifluoromethyl)cyclohex-1-enyl)-3-nitropyridine (1.0 eq.) in ethanol (0.13

M) was blown with the help of M" for 20 minutes. Then, Bos anhydride (1.5 equiv.) and Ra/C (0.2 equiv.) were added to the reaction mixture. The reaction mixture was stirred at room temperature under a He atmosphere overnight. The solid was removed by filtration through celite and washed with EN.

The residue was purified on a column (595 methanol in a 1:11 mixture of ethyl acetate and hexanes) and racemic cis-3-(3-aminopyridin-4-yl)-5-(trifluoromethyl)cyclohexylcarbamate (57 95) was obtained. LC/MS (ti/lg):

MN. - 360.2, -0.72 min. Enantiomerically pure tert-butyl-(1A8, ZA, 55)-3-(3-aminopyridin-4-yl)-5-(trifluoromethyl)cyclohexylcarbamate and M-(4-((15, 35, 5A)-3-amino -5-(trifluoromethyl)cyclohexyl)pyridin-3-yl)-6-(2,6-difluorophenyl)-5-fluoropicolinamide was isolated by chiral HPLC (for analysis Ke-8.14 min and 10.59 min, respectively; heptane :isopropanol-- 90:10 (vol.:06.), SpigaIse! IS 100 (4.6 mm at 1 ml/min. For preparative separation, heptane:isopropanol - 90:10 (vol.:06.), SpigaiseI IC 250 (20 mm at 15 ml/min).

Synthesis of (K)-4-benzyl-3-(2B, 3H)-3-(H)-2,2-dimethyl-1,3-dioxolan-4-yl)-3-hydroxy-2-methylpropanoyl)oxazolidin- 2nd point

M

(F) IF)

M not (8)-4-benzyl-3-(2B, 3H)-3-(H)-2,2-dimethyl-1,3-dioxolan-4-yl)-3-hydroxy-2-methylpropanoyl)oxazolidine -2-one was obtained according to the method (Rhos.Mai.Asai.Bsieepse5, 101, 33, 2004, pp. 12042-12047) described for the enantiomeric compound using (K)-4-benzyl-Z-propionyloxazolidine- 2-one and acetonide of E-glyceraldehyde.

Synthesis of (K)-4-benzyl-3-(28, C3A)-3-(tert-butyldimethylsilyloxy)-3-(H)-2,2-dimethyl-1,3-dioxolan-4-yl)y-2 -methylpropanoyl)oxazolidin-2-one

X Heh

M

(F) |c)

MA, Fr. (8)-4-benzyl-3-(28,3НА)-3-(tert-butyldimethylsilyloxy)-3-(H)-2,2-dimethyl-1,3-dioxolan-4-yl)-2-methylpropanoyl )oxazolidin-2-one was obtained according to the method (Rhos.Mai.Asai.Bsieepse5, 101, 33, 2004, pp. 12042-12047) described for the enantiomeric compound using (K)-4-benzyl-3- (28, 3H)-3-(B)-2,2-dimethyl-1,3-dioxolan-4-yl)-3-hydroxy-2-methylpropanoyl)oxazolidin-2-one.

Synthesis of (25, 3K)-3-(tert-butyldimethylsilyloxy)-3-((B)-2,2-dimethyl-1,3-dioxolan-4-yl)-2-methylpropan-1-ol

There is no answer

To the solution /(K)-4-benzyl-3-(28, C3B)-3-(tert-butyldimethylsilyloxy)-3-((B8)-2,2-dimethyl-1,3-dioxolan-4-yl) -2-methylpropanoyl)oxazolidin-2-one (1.0 equiv.) and ethanol (3.0 equiv.) in THF (0.09 M) at - 40 "C was added I IVNa (1.0 equiv.). The reaction mixture was allowed to slowly warm to room temperature and stirred at this temperature for 12 h. The solution was recooled to -40 °C and additional GiIVNa (0.3 equiv.) was added. After reheating to RT and stirring for 2 h. the solution was diluted with diethyl ether and added 1 n. Mahon). The resulting mixture was extracted with ethyl acetate, the organic layer was separated, washed with Massi(za), dried over magnesium sulfate, filtered and concentrated. The residue was purified using column chromatography on silica gel (10-30 Fo

EMTOAc/ n-heptanes) and obtained (25, C3K)-3-(tert-butyldimethylsilyloxy)-3-((B)-2,2-dimethyl-1,3-dioxolan-4-yl)-2-methylpropan- 1-ol in (75 95). LC/MS - 2471 (MaN-ketal-NgO), He-0.64 min.

Synthesis of (Kk, 25)-3-azido-1-((H)-2,2-dimethyl-1,3-dioxolan-4-yl)-2-methylpropoxy)(tert-butyl)dimethylsilane

Отв5 tuyu oh,

To a solution of (25, 3K)-3-(tert-butyldimethylsilyloxy)-3-((B)-2,2-dimethyl-1,3-dioxolan-4-yl)-2-methylpropan-1-ol (1, 0 equiv.), DIAD (diisopropyl azodicarboxylate) (2.0 equiv.) and PPIz (2.0 equiv.) in THF (0.18 M) was added with DFFA (diphenylphosphorylazide) (1.0 equiv., 1 M solution in THF) . The reaction mixture was stirred at room temperature overnight. After removal of volatile substances in a vacuum, the residue was purified by column chromatography on silica gel (2-3-5 96 EOASc/ n-heptanes) and obtained (Kk, 25)-3-azido-1-((H)-2,2- dimethyl-1,3-dioxolan-4-yl)-2-methylpropoxy)(tert-butyl)dimethylsilane (62 95). "H-NMR (400 MHz, COsSys): 5 4.04-4.10 (t, 1H), 3.94 (aa, 9U-8.0, 6.4, 1H), 3.72 (a, y-7.2, 1H), 3.53 (I, 9U-8.0, 1H), 3.36 (aa, U-12, 8.0, 1H), 3.19 (aa, 9U-12 ,0, 6.7, 1H), 1.52-1.60 (t, 1H), 1.41 (5, ЗН), 1.34 (5, ЗН), 0.92 (9, 9-72 , ЗН), 0.90 (5, 9 Н), 0.12 (5, ЗН), 0.09 (5, ЗН).

Synthesis of (2E,, ZK, 45)-5-azido-3-(tert-butyldimethylsilyloxy)-4-methylpentane-1 2-diol

ОтТв5 throne on

To a solution of ((1K, 25)-3-azido-1-(H)-2,2-dimethyl-1,3-dioxolan-4-yl)-2-methylpropoxy)(tert-butyl)dimethylsilane (1.0 eq.) in MeOH (0.1 M) was added PPTS (pyridine-p-toluenesulfonate) (1.0 eq.) and the mixture was stirred at RT for 14 h, at 50 "C for 2 h and at 80 "C within 1 hour Volatiles were removed in vacuo and the residue was purified by column chromatography on silica gel (10-2595 EtOAc/n-heptanes) to give (2K, ZK, 45)-5-azido-3-(tert-butyldimethylsilyloxy)-4-methylpentane- 1,2-diol (40 Ob).

Synthesis of (2K, ZK, 45)-5-azido-3-(tert-butyldimethylsilyloxy)-2-hydroxy-4-methylpentyl /-4-methylbenzenesulfonate

ОтТв5 чтрттота он

To a solution of tert-butyl-(28, ZB, 45)-5-azido-3-(tert-butyldimethylsilyloxy)-1-hydroxy-4-methylpentan-2-ylcarbamate (1.0 equiv.) in pyridine (0.2 M) was added RT5SiI (1.3 eq.) at 0 "C. The mixture was kept at the same temperature for 16 h. Volatile substances were removed in vacuo and the residue was purified by column chromatography on silica gel (10-15 2095 EUAbs/n-heptanes ) and obtained (2, ZK, 45)-5-azido-3-(tert-butyldimethylsilyloxy)-2-hydroxy-4-methylpentyl /-4-methylbenzenesulfonate.

Synthesis of (2K, ZE, 45)-5-azido-2,3-bis(tert-butyldimethylsilyloxy)-4-methylpentyl 4-methylbenzenesulfonate

ОтТв5 tr тртот отв

To a solution of (2, ZK, 45)-5-azido-3-"(tert-butyldimethylsilyloxy)-2-hydroxy-4-methylpentyl 4-methylbenzenesulfonate (1.0 equiv.) and 2,6-lutidine (3.4 eq.) at 0 "С, TVOM5OTI (1.7 eq.) was added.

The solution was stirred for 7 h while it was heated to RT. The solution was diluted with

EtOAc, washed with 1095 Si5O», H2O, Ma»zhSO3(za), MaSivzau, dried over M4oazoa, filtered and concentrated. The residue was purified by column chromatography on silica gel (2,5-5-10-20 95 EtOAc/ n-heptanes) and obtained (2, ZK, 45)-5-azido-2,3-bis(tert-butyldimethylsilyloxy)- 4-methylpentyl 4-methylbenzenesulfonate (75 9).

Synthesis of 4-(ЗВ, AB, 55)-3,4-bis(tert-butyldimethylsilyloxy)-5-methylpiperidin-1-yl)-3-nitropyridine

Otv5 bSru

M what

M

A solution of (2k, ZE, 45)-5-azido-2,3-bis(tert-butyldimethylsilyloxy)-4-methylpentyl 4-methylbenzenesulfonate in EN (0.05 M) was degassed with argon. DIEA (1.5 eq.) was added, followed by 10 95

Ra/C (0.1 equiv.). The reaction mixture for 3 hours. stirred in the atmosphere of hydrogen supplied from a cylinder. The solution was degassed and purged with argon, and then 4-chloro-3-nitropyridine (1.5 eq.) was added and further

DIEA (1.5 equiv.). After stirring at CT for 15 h. the solution was filtered to remove Ra/C and the volatiles were removed in vacuo. The residue was diluted with ethyl acetate and washed with

MagSO3(zayu, MasSiza", dried over Mo933O", filtered and concentrated. The residue was purified using column chromatography on silica gel (10-15 96 E(OAc/ n-heptanes) and obtained 4-((ЗВ,

AB, 55)-3,4-bis(tert-butyldimethylsilyloxy)-o-methylpiperidin-1-yl)-3-nitropyridine (40 95). PH/MS - 482.4 (MAN), Ne1.26 min.

Synthesis of 4-(ЗНВ, 48, 55)-3,4-bis(tert-butyldimethylsilyloxy)-5-methylpiperidin-1-yl)pyridin-3-amine

Answer that

M

There are more

M

To a solution of 4-(Z3А8, 48, 55)-3,4-bis(tert-butyldimethylsilyloxy)-5-methylpiperidin-1-yl)-3-nitropyridine (1.0 equiv.) in ethanol at a concentration equal to 0, 05 M, 10 95 palladium on charcoal (0.1 equiv.) was added. The obtained heterogeneous solution was placed in a hydrogen atmosphere and stirred for 14 hours. Then the mixture was filtered through a layer of celite eluting with ethanol. Volatiles were removed in vacuo to give 4-(3B8, 48, 55)-3,4-bis(tert-butyldimethylsilyloxy)-5-methylpiperidin-1-yl)upyridin-3-amine. LC/MS - 452.4 (MN), K-1.31 min.

Synthesis of (K)-tert-butyl-4-((1B8, 2H8)-3-((H)-4-benzyl-2-oxooxazilidin-3-yl)-1-hydroxy-2-methyl-3-oxopropyl) -2,2-dimethyloxazilidine-3-carboxylate shah

M

M, wax

To a solution of (K)-4-benzyl-3-propionyloxazolidin-2-one (1.0 equiv.) in DCM (0.13 M) at -407C was added TisSi (1.0 equiv.). The mixture was stirred at -40 "C for 10 min (yellow suspension), then DIPEA (2.5 equiv.) (dark red solution) was added and stirred at 0 "C for 20 min. Then (K)-tert-butyl-4-formyl-2,2-dimethyloxazilidine-3-carboxylate (1.0 equiv.) in DCM (0.5 M) was added dropwise and the resulting mixture was stirred for 1.5 h. The reaction was stopped by adding an aqueous solution of ammonium chloride and the mixture was extracted with ethyl acetate. The organic phase was separated, washed with brine, dried over magnesium sulfate, filtered and concentrated. The residue was purified by column chromatography eluting with ethyl acetate and hexanes (1:4) to obtain (K)-tert-butyl-4-((18, 28)-3-(B)-4-benzyl-2-oxooxazilidine-3 -yl)-1-hydroxy-2-methyl-3-oxopropyl)-2,2-dimethyloxazilidine-Z-carboxylate as the main product (5:2) with a yield of 58 95. LC/MS - 363.3 (Ma-H -Vos), KCh-1.09 min.

Synthesis of (B)-tert-butyl-4-(1B, 28)-3-((8)-4-benzyl-2-oxooxazilidin-3-yl)-1-(tert-butyldimethylsilyloxy)-2-methyl-3 -oxopropyl)-2,2-dimethyloxazilidine-3-carboxylate

In OotTV5

M

IF) ru

MA Wes,

To the solution (K)-tert-butyl-4-(1B8, 2H8)-3-(B8)-4-benzyl-2-oxooxazilidin-3-yl)-1-hydroxy-2-methyl-

3-oxopropyl)-2,2-dimethyloxazilidine-3-carboxylate (1.0 equiv.) and lutidine (1.8 equiv.) in DCM (0.1 M) at -40 "C was added TV5OTI (1.4 equiv. ). The reaction mixture was stirred at -40 "C for 2 hours. The solution was diluted with ethyl acetate and washed with a saturated solution of MnSO 2 , a saturated solution of Masi, dried over magnesium sulfate, filtered and concentrated. The residue was purified by column chromatography on silica gel eluting with ethyl acetate and hexanes (1:4) and (B)-tert-butyl-4-(18, 28)-3-((8)-4-benzyl-2-oxooxazilidine) was obtained -3-yl)-1-(tert-butyldimethylsilyloxy)-2-methyl-3-oxopropyl)-2,2-dimethyloxazilidine-3-carboxylate as the main product (5:2) in 83 95 yield.

LC/MS - 577.3 (MH), EC - 1.33 min (methodology with selection of fractions 65-95 95).

Synthesis of (K)-tert-butyl-4-((18A, 25)-1-(tert-butyldimethylsilyloxy)-3-hydroxy-2-methylpropyl)-2,2-dimethyloxazilidine-3-carboxylate

Отв5 nut

weight,

To the solution (K)-tert-butyl-4-(1B, 2I8)-3-((8)-4-benzyl-2-oxooxazilidin-3-yl)-1-(tert-butyldimethylsilyloxy)-2-methyl- 3-oxopropyl)-2,2-dimethyloxazilidine-3-carboxylate (1.0 equiv.) and ethanol (3.0 equiv.) in THF (0.09 M) at -30 "C was added GiVNa (3.0 equiv. .). The reaction mixture was allowed to warm up to 0 C and was stirred at this temperature for 3 hours. Then the solution was diluted with diethyl ether and 1 N Mn was added. The resulting mixture was extracted with ethyl acetate, the organic layer was separated, washed with a saturated solution of MgSi, dried over magnesium sulfate, The residue was purified by silica gel column chromatography eluting with ethyl acetate and hexanes (1:4) to give (K)-tert-butyl-4-(1B, 25)-1-(tert-butyldimethylsilyloxy)-3- hydroxy-2-methylpropyl)-2,2-dimethyloxazilidine-3-carboxylate as the main product (ratio 5:22) with a yield of 7195. LC/MS - 304.3 (Mae-H-Boc), K-0.95 min (methodology with selection of fractions 65-95 9).

Synthesis of (K)-tert-butyl-4-(18, 25)-3-azido-1-(tert-butyldimethylsilyloxy)-2-methylpropyl)-2,2-dimethyloxazilidine-3-carboxylate

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weight,

To a solution of (K)-tert-butyl-4-(18, 25)-1-(tert-butyldimethylsilyloxy)-3-hydroxy-2-methylpropyl)-2,2-dimethyloxazilidine-3-carboxylate (1.0 equiv. ), DIAD (2.0 equiv.) and PPAz (2.0 equiv.) in THF (0.18 M) were added DFFA (2.0 equiv., 1 M solution in THF). The reaction mixture was stirred at room temperature overnight. After removal of volatile substances in a vacuum, the residue was purified by column chromatography on silica gel eluting with ethyl acetate and hexanes (1:6) and obtained (K)-tert-butyl-4-(1А8, 25)-3-azido-1-(tert -butyldimethylsilyloxy)-2-methylpropyl)-2,2-dimethyloxazilidine-3-carboxylate as the main product (5:2) with a yield of 8695. LC/MS - 329.3 (Ma-H-Boc), H-1.40 min (methodology with selection of fractions 65-95 9).

Synthesis of tert-butyl-(28, ZA, 45)-5-azido-3-(tert-butyldimethylsilyloxy)-1-hydroxy-4-methylpentan-2-ylcarbamate

Отв5 pana

Ministry of Higher Education

To a solution of (K)-tert-butyl-4-((18, 25)-3-azido-1-(tert-butyldimethylsilyloxy)-2-methylpropyl)-2,2-dimethyloxazilidine-3-carboxylate (1.0 equiv .) in EN (0.1 M) was added PPTS (1.3 equiv.) and the mixture was refluxed for 2 days. Volatile substances were removed in vacuo, the residue was dissolved in DCM (0.1 M) and DIEA (1.5 equiv.) and VosgO (1.0 equiv.) was added to the reaction mixture. The solution was stirred for three hours. at room temperature. Solvents were removed under reduced pressure and the residue was diluted with ethyl acetate, washed with water, aqueous Manzo solution, aqueous

Manso", saturated Masi solution. The organic phase was dried over magnesium sulfate, filtered and concentrated. The residue was purified by column chromatography on silica gel eluting with ethyl acetate and hexanes (1:3) to obtain tert-butyl-(2А8, ЗА, 45)-5-azido-3-(tert-butyldimethylsilyloxy)-1-hydroxy-4- methylpentan-2-ylcarbamate as the main isomer (5:2) with a yield of 70 95. LC/MS - 289.3 (MaH-Vos), EC-0.76 min (methodology with selection of fractions 65-95 Ob).

Synthesis of (2K, ZK, 45)-5-azido-2-(tert-butoxycarbonylamino)-3-(tert-butyldimethylsilyloxy)-4-methylpentylmethanesulfonate

Отв5 pana

Ministry of Higher Education

To a solution of tert-butyl-(28, ZB, 45)-5-azido-3-(tert-butyldimethylsilyloxy)-1-hydroxy-4-methylpentan-2-ylcarbamate (1.0 equiv.) in pyridine (0.2 M) at 0 "C, M5ClII (1.3 equiv.), then DMAP (catalytic amount) was added. The mixture was stirred at this temperature for 1 hour. The solution was diluted with ether and ethyl acetate (4:11), washed with an aqueous solution of ManzoO»x, saturated Mansoz solution, brine, dried over magnesium sulfate, filtered and concentrated. butoxycarbonylamino)-3-"(tert-butyldimethylsilyloxy)-4-

methylpentyl methanesulfonate as the main isomer (5:2) with a yield of 90 90. LC/MS - 367.3 (Ma-H-Bocs),

K-O, 81 min (methodology with the selection of fractions 65-95 9).

Synthesis of tert-butyl-(ZB, 48, 55)-4-(tert-butyldimethylsilyloxy)-5-methylpiperidin-3-ylcarbamate

Ootv5 -tbeu i

A solution of (2K, ZK, 45)-5-azido-2-(tert-butoxycarbonylamino)-3-tert-butyldimethylsilyloxy)-4-methylpentylmethanesulfonate in Meon (0.09 M) was degassed with nitrogen for 20 min. DIEA (2.5 eq.) was added, followed by 10 95 Ra/C (0.1 eq.). The reaction mixture was stirred for 2 hours. in an atmosphere of hydrogen supplied from a cylinder. The solution was filtered and the filtrate was concentrated in vacuo to give tert-butyl-(ZB, 48, 55)-4-(tert-butyldimethylsilyloxy)-5-methylpiperidin-3-ylcarbamate as the major isomer (5:2) with a yield of 299 95. РХ /MS - 345.2 (Ma-N-Vos), CI-0.95 and 0.99 min.

Synthesis of tert-butyl-(ZA, 48, 55)-4-(tert-butyldimethylsilyloxy)-5-methyl-1-(3-nitropyridin-4-ylpiperidin-3-ylcarbamate)

Ootv5 -teu

M

ZMO" more

M

DIEA ( 2.5 equiv.) and 4-chloro-3-nitropyridine (1.5 equiv.).

The reaction mixture was stirred at 60 "C for 2 h. Volatile substances were removed in vacuo, the residue was diluted with ethyl acetate and washed with saturated Massey solution. The organic phase was dried over magnesium sulfate, filtered and concentrated. The crude substance was purified by column chromatography on silica gel eluting with ethyl acetate and hexanes (1:2) and obtained tert-butyl-(ZA8, 48, 55)-4-(tert-butyldimethylsilyloxy)-5-methyl-1-(3-nitropyridin-4-yl)piperidin-3-ylcarbamate with yield 76 95. RH/MS - 467.3 (MAN), K-1.09 min.

Synthesis of tert-butyl-(ZA, 48, /55)-1-(3-aminopyridin-4-yl)-4-tert-butyldimethylsilyloxy)-5-methylpiperidin-3-ylcarbamate

Ootv5 -tjeu

M from MN" more

M

A solution of tert-butyl-(ZA, 48, 55)-4-(tert-butyldimethylsilyloxy)-5-methyl-1-(3-nitropyridin-4-ylpiperidin-3-ylcarbamate) (1.0 equiv.) in MeOH (0 .05 M) was degassed with nitrogen for 20 min. 1095 Pa/C (0.2 equiv.) was added to the mixture and the solution was stirred for 3 h in an atmosphere of hydrogen supplied from a cylinder. The reaction mixture was filtered and the filtrate was concentrated under reduced pressure and obtained tert-butyl-(ZA, 48, /55)-1-(3-aminopyridin-4-yl)-4-(tert-butyldimethylsilyloxy)-5-methylpiperidin-3-ylcarbamate as the desired product with a yield of 94 95. РХ / MS - 437.4 (MAN), Ch-1.08 min. "H-NMR (300 MHz, SOS"): 6 8.01 (in, 1H), 7.95 (a, 9-6.0 Gu, 1H), 6.76 (a, U-6.0 Hz, 1H), 4.44 (g 5, 1H), 3.74 (horn 5, 2H), 3.59-3.55 (t , 1H), 3.25-3.13 (t, 2H), 2.47-2.35 (t, 2H), 1.89 (reg 5, 2H), 1.44 (5, 9H), 1 .04 (а, 9-6.0, ЗН), 0.92 (5, 9Н), 0.13 (а, 9У-9.0, 6Н).

Synthesis of tert-butyl-(28)-1-(benzyloxy)-3-hydroxy-4-methylhex-5-en-2-ylcarbamate

N N

M u and sov

(72)-2-(but-2-enyl) was added to a solution of M-Boc-O-benzyl-Y-serine aldehyde (1.0 equiv.) in DCM (0.1 M) at -78"7С in an Ar atmosphere )-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (1.1 equiv) and the clear solution was stirred for 16 h while it warmed to RT.The solution was added to EtOAc and washed with H2O (Zh) and MaSyza)", dried over Mo9a5O4 and purified by chromatography on silica gel (15 95 EAs/hexanes) and according to "H NMR data, tert-butyl-(28)-1-(benzyloxy)-3-hydroxy- 4-methylhex-5-en-2-ylcarbamate (54 95) in the form of a mixture of isomers with a composition of 3:1.

RHMS (t/g2): 236.3 (MH"-Voc); RH Ki-4.37 and 4.51 min.

Synthesis of (4K)-4-(benzyloxymethyl)-5-(but-3-en-2-yl)uoxazolidin-2-one c) 2-4 - ОВп

To a solution of (2H8)-1-(benzyloxy)-3-hydroxy-4-methylhex-5-en-2- in THF (0.1 M) was added 60 95 sodium hydride in mineral oil (1.5 equiv.). After stirring for 3 days, the reaction was stopped by adding MNaCl(a) and the solution was diluted with EtOAc and washed with MNaCl(a)) and

MaSi(za), dried over Mo95O5 and purified by chromatography on silica gel (50 95

EtOAc/hexanes) and obtained (4H)-4-(benzyloxymethyl)-5-(but-3-en-2-yl)uoxazolidin-2-one (89 95)..-in the form of a mixture of composition 3:1. RHMS (t/2): 262.2 (MNU); РХ Ke3.47 min.

Synthesis of (48)-4-(benzyloxymethyl)-5-(1-hydroxypropan-2-yl)uoxazolidin-2-one in) 24 no OVp

To a solution of (4H8)-4-(benzyloxymethyl)-5-(but-3-en-2-yl)uoxazolidin-2-one (1.0 equiv.) in a 2:1 mixture

Meon/No (0.04 M) was added with 4 95 osmium tetroxide in HgO (0.07 equiv.) and sodium periodate (3.0 equiv.).

After stirring for three hours. the white precipitate was filtered off and washed with EtOAc.

The combined filtrate was concentrated in vacuo and the residue was dissolved in EIOAc, washed with

Mass(za), dried over Ma5O»5, filtered and concentrated. The crude aldehyde was dissolved in EOH (0.08 M) and sodium borohydride (2.0 equiv.) was added upon cooling to 0 °C. After stirring for 15 h and heating to room temperature, the reaction was stopped by adding H2O.

After stirring for 20 min, EN was removed under vacuum, EIOAc was added, and the solution was washed with 1 n. NCI, MaHSO3(zaz and MaSyza), dried over MdoZO", filtered and concentrated, and after purification by chromatography on silica gel, (4H)-4-(benzyloxymethyl)-5-(1-hydroxypropan-2-yl)uoxazolidin-2 was obtained -one in the form of a mixture of isomers of composition 3:1 (60 95). RHMS (t/h2): 266.1 (MNU); РХ 22.28 min.

Synthesis of (4H8)-4-(hydroxymethyl)-5-(1-hydroxypropan-2-yl)uoxazolidin-2-one in) o-4 no OH

10 95 palladium was added to a solution of (4H)-4-(benzyloxymethyl)-5-(1-hydroxypropan-2-yl)uoxazolidin-2-one (1.0 equiv.) in methanol at a concentration equal to 0.1 M on coal (0.1 equiv.). The obtained heterogeneous solution was placed in a hydrogen atmosphere and stirred for 15 hours. Then the mixture was filtered through a layer of celite eluting with methanol. Volatiles were removed in vacuo to give (4H)-4-(hydroxymethyl)-5-(1-hydroxypropan-2-yl)uoxazolidin-2-one (9995). RHMS (t/y): 176.1 (MNU).

Synthesis of 2-(48)-2-oxo-4-(tosyloxymethyl)oxazolidin-5-yl)-propyl 4-methylbenzenesulfonate c) 2-4 tvo Ot

To a solution of (4H)-4-(hydroxymethyl)-5-(1-hydroxypropan-2-yl)uoxazolidin-2-one (1.0 equiv.) in pyridine

(0.15 M) at 0 "С, p-toluenesulfonyl chloride (2.1 equiv.) was added. The solution was allowed to warm to RT with stirring for 14 h, and then E(Ac) was added and the solution was washed with HgO (Zh),

SibOdvats (2x), HgO, MgSO2(za" and MaSi(zao, dried over Md5O", filtered, concentrated and purified using chromatography on silica gel (75 90EOAs/hexanes eluent) and obtained 2-((4H)-2-oxo -4-(tosyloxymethyl)oxazolidin-5-yl/upropyl 4-methylbenzenesulfonate (68 95) LCMS (t/g): 4841 (MNU);

RH G-4.06 min.

Synthesis of (Zav, 7B, 7ab)-5-(4-methoxybenzyl)-7-methylhexahydrooxazolo|4,5-s|Ipyridin-2(ZH)-one and (Zav, 75, 7an)-5-(4-methoxybenzyl) )-7-methylhexahydrooxazolo|4,5-s|Ipyridin-2(ZH)-one (c) 0-4 0-Х и и

RMV RMV

A solution of 2-((48)-2-oxo-4-(tosyloxymethyl)oxazolidin-5-ylylpropyl. 4-methylbenzenesulfonate (1.0 eq.), diisopropylethylamine (3.0 eq.) and para-methoxybenzylamine (1.5 equiv.) in MMR (0.05 M) was heated at 100 "C for 14 h. The solution was immediately purified by ZF (reversed phase) HPLC. The product fractions were desalted by addition to EIOAc and MgCO3(eg), additionally washed

MaSi(za), dried over Ma5oO" and concentrated and obtained 2 separate isomers, (Zak, 7K, 7az)-5-(4- methoxybenzyl)-7-methylhexahydrooxazolo|4,5-c|Ipyridine-2(ZH)- one and (zac, 15, tatk)-5-(4-methoxybenzyl)-7-methylhexahydrooxazolo|4,5-c|Ipyridin-2(ZH)-one (27 95 and 895). RHMS (t/2): 277.2 (MNU) at 0.40 and 0.42 min.

Synthesis of (Zav, 7B, 7a5)-7-methylhexahydrooxazolo|4,5-s|Ipyridin-2(ZH)-one 0-4

M

N

To a solution of (Zary, 7B, 7ab5)-5-(4-methoxybenzyl)-7-methylhexahydrooxazolo|4,5-s|Ipyridin-2(ZH)-one (1.0 equiv.) in methanol at a concentration equal to 0 .1 M, 20 95 palladium hydroxide on charcoal (0.3 equiv.) was added. The obtained heterogeneous solution was placed in a hydrogen atmosphere and stirred for 2 hours.

Then the mixture was filtered through a layer of celite eluting with methanol. Volatile substances were removed in vacuo and (Zav, 7B, 7авб5)-7-methylhexahydrooxazolo!|4,5-с|Ipyridin-2(ZH)-one (99 95) was obtained. RHMS (t/2): 157.1 (MNU) at 0.16 min.

Synthesis of (Zak, 7K, 7avz)-tert-butyl-7-methyl-5-(3-nitropyridin-4-yl)-2-oxohexahydrooxazolo|4,5-sipyridine-3(2H)-carboxylate (c) 9- X ru

M d MO" schi

M

A solution of 4-chloro-3-nitropyridine (1.3 equiv.) and (Zav, 7B, 7ab5)-7-methylhexahydrooxazolo!|4,5-c|Ipyridin-2(ZN)-one (1.5 equiv.) in СНоСі" at a concentration equal to 0.1 M, was stirred at CT for 48 hours. and then piperidine (0.4 eq.) was added to consume excess 4-chloro-3-nitropyridine. After stirring for another 2 hours. di-tert-butyldicarbonate (2.0 equiv.) and dimethylaminopyridine (0.1 equiv.) were added. After stirring for 4 hours. the solution was subjected to distribution between

EUAs and Mansoz (za, additionally washed with Mansoz (za) and MaSivao, DRIED over Mao, filtered and purified by silica gel chromatography and obtained (Zav, 7B, 7a5)-tert-butyl-7-methyl-5-( 3-Nitropyridin-4-yl)-2-oxohexahydrooxazolo|4,5-s|Ipyridine-3(2H)carboxylate (62 Ob).

RHMS (t/g2): 379.0 (MNU) at 0.58 min.

Synthesis of (Zai, 7B, 7ab)-tert-butyl-5-(3-aminopyridin-4-yl)-7-methyl-2-oxohexahydrooxazolo|4,5-

Cypyridine-Z(2H)-carboxylate c) 9-5 ru

More

M

To a solution of (zan, 7B, Ttav5)-tert-butyl-7-methyl-5-(3-nitropyridin-4-yl)-2-oxohexahydrooxazolo!|4,5-s|Ipyridine-3(2H)-carboxylate ( 1.0 equiv.) in methanol at a concentration equal to 0.1 M, 10 95 palladium on carbon (0.1 equiv.) was added. The obtained heterogeneous solution was placed in a hydrogen atmosphere and stirred for 14 hours. Then the mixture was filtered through a layer of celite eluting with methanol. The volatile substances were removed in vacuo and obtained (Zav, 7B, 7ab5)-tert-butyl-5-(3-aminopyridin-4-yl)-7-methyl-2-oxohexahydrooxazolo|4,5-c|Ipyridine-3(2H )-carboxylate. RHMS (t/y): 3491 (MNU); РХ h-2.06 min.

Synthesis of (zak, 75, 7ak)-7-methylhexahydrooxazolo|4,5-s|Ipyridin-2(ZH)-one 0-6 "su

M

N

To a solution of (Zan, 75, 7an)-5-(4-methoxybenzyl)-7-methylhexahydrooxazolo|4,5-s|Ipyridin-2(ZH)-one (1.0 equiv.) in methanol at a concentration equal to 0 .1 M, 20 95 palladium hydroxide on charcoal (0.3 equiv.) was added. The obtained heterogeneous solution was placed in a hydrogen atmosphere and stirred for 2 hours.

Then the mixture was filtered through a layer of celite eluting with methanol. Volatile substances were removed in vacuo and (Zan, 75, 7an)-7-methylhexahydrooxazolo!|4,5-c|Ipyridin-2(ZH)-one (99 95) was obtained. RHMS (t/2): 157.1 (MNU) at 0.17 min.

Synthesis of (Zak, 75, 7ak)-tert-butyl-7-methyl-5-(3-nitropyridin-4-yl)-2-oxohexahydrooxazolo|4,5-sipyridine-3(2H)-carboxylate (c) 2- 4 "ears

More

M

A solution of 4-chloro-3-nitropyridine (1.3 equiv.) and (Zav, 75, 7an)-7-methylhexahydrooxazolo!|4,5-c|Ipyridin-2(ZN)-one (1.5 equiv.) in СНоСі" at a concentration equal to 0.1 M, was stirred at RT for 48 h, then piperidine (0.4 equiv.) was added to consume excess 4-chloro-3-nitropyridine. After stirring for another 2 hours. di-tert-butyldicarbonate (2.0 equiv.) and dimethylaminopyridine (0.1 equiv.) were added. After stirring for 4 hours. the solution was subjected to distribution between

EuAc and Mansoz (za, additionally washed with Mansoz (za) and MaSivao, DRIED over Mao, filtered and purified by chromatography on silica gel (eluent 75 95 EOAc/hexanes) and obtained (Zai, 75, 7an)-tert-butyl -7-methyl-5-(3-nitropyridin-4-yl)-2-oxohexahydrooxazolo|4,5-sipyridine-3(2H)-carboxylate (35 95). LCMS (t/2): 379.0 (MNU ).RH Ke2.42 min.

Synthesis of (Zak, 7K, 7az)-tert-butyl-5-(3-aminopyridin-4-yl)-7-methyl-2-oxohexahydrooxazolo|4,5-sipyridine-3(2H)-carboxylate c) 2-4 "uh

More

M

To a solution of (zan, 75, tav)-tert-butyl-7-methyl-5-(3-nitropyridin-4-yl)-2-oxohexahydrooxazol!|4,5-c|Ipyridine-3(2H)-carboxylate ( 1.0 equiv.) in methanol at a concentration equal to 0.1 M, 10 95 palladium on carbon (0.1 equiv.) was added. The obtained heterogeneous solution was placed in a hydrogen atmosphere and stirred for 14 hours. Then the mixture was filtered through a layer of celite eluting with methanol. Volatiles were removed in vacuo to give (Zan, 75, 7an)-tert-butyl-5-(3Z-aminopyridin-4-yl)-7-methyl-2-oxohexahydrooxazolo!|4,5-c|Ipyridine-Z( 2H)-carboxylate. RHMS (t/2): 349.1 (MNU; RH Ch-2.18 min.

Methodology 1

Synthesis of methyl-33-amino-6-(2,6-difluorophenyl)picolinate

E E ro, Zh

MN"

A solution of methyl 3-amino-6-bromopicolinate (1.0 equiv.), 2,6-difluorophenylboronic acid (3.0 equiv.) and

Ra(arry)CI--DHM (0.1 equiv.) in a mixture of 31 DME/2M Mag2bOz (0.5 M) was heated by microwave radiation at 120 "C with 15-minute intervals. The reaction mixture was filtered and washed with EOAc. The organics were partitioned with H2O (25 mL), further washed with NaCl (25 mL), dried over Md 5 O, and the volatiles removed in vacuo. The residue was diluted with EOAc and passed through a pad of silica gel and the volatiles were removed in vacuum and obtained methyl-3-amino-6-(2,6-difluorophenyl)picolinate (47 90).LCMS (t/2): 265.1 (MNU);

РХ e-2.70 min

Synthesis of 6-(2,3-difluorophenyl)-5-fluoropicolinic acid

IS

What are we? - c)

2,3-Difluorophenylboronic acid (1.3 equiv) was added to a solution of b-bromo-5-fluoropicolinic acid (1.0 equiv.) in DME and 2M MgSO3 (3:1, 0.25 M) in a microwave oven vessel. ) and Pa(arrOCl»--DXM (0.05 equiv.). The vessel was heated in a microwave oven at 120 "C for 30 min. The mixture was diluted with ethyl acetate and 1 N. Mn was added. The organic phase was separated and extracted three more times with 1N NaOH and 1 time with bN NaOH. The combined aqueous phases were filtered and acidified to pH 1 by addition of conc. HCl and extracted with ethyl acetate. The organic layer was dried over magnesium sulfate, filtered and concentrated to give 6-(2, 3-difluorophenyl)-

B-fluoropicolinic acid with a yield of 78 95. РХ/МСО - 254.1 (MAN), Ch-0.75 min.

Methodology 2

Synthesis of 3-amino-6-(-2,6-difluorophenyl)picolinic acid

E E

Well, well

MN"

To a solution of methyl-3-amino-6-(-2,6-difluorophenyl)picolinate (1.0 equiv.) in THF (0.5 M) was added 1M

MON (4.0 eq.). After stirring for 4 hours. at 60 "C, 1N HCl (4.0 equiv.) was added and THF was removed in vacuo. The resulting solid was filtered and washed with cold H2O (3x20 mol) to give 3-amino-6-(2,6-difluorophenyl) picolinic acid (90 95). LCMS (t/2): 251.1 (MNU); LC Ke-2.1

MIN.

Synthesis of 3-amino-6-(2-fluoro-5-propoxyphenyl)picolinic acid

IS

MO schi) t o Mne"

According to method 1 using 33-amino-6-bromopicolinic acid (1.0 equiv.) and 2-fluoro-5-propoxyphenylboronic acid (1.5 equiv.) and Ra(arrOSI-A-DCM (0.05 equiv. ) obtained 3-amino-6-(2-fluoro-5-propoxyphenyl)picolinic acid with a yield of 75 90. LC/MS "- 291.0 (MH), K-0.81 min.

Synthesis of 3-amino-5-fluoro-6-(2-fluoro-5-propoxyphenyl)picolinic acid

Ot

E we shchi) -7b o Mn,

According to method 1 using 3-amino-6-bromo-5-fluoropicolinic acid (1.0 equiv.) and 2-fluoro-5-propoxyphenylboronic acid (1.3 equiv.) .05 equiv.) 3-amino-5-fluoro-6-(2-fluoro-5-propoxyphenyl)picolinic acid was obtained with a yield of 28 90. LC/MS - 309.1 (Ma-H), K-1.00 min.

Synthesis of methyl-3-amino-5-fluoro-6-(2-fluorophenyl)picolinate

E shk and c) - oh Me,

According to method 1 using methyl-3-amino-6-bromo-5-fluoropicolinate (1.0 equiv.) and 2-fluorophenylboronic acid (1.5 equiv.) .) obtained methyl-3-amino-5-fluoro-6-(2-fluorophenyl)picolinate with a yield of 299 95. LC/MS - 265.0 (MH), Ch-0.77 min.

Synthesis of 3-amino-5-fluoro-6-(2-fluorophenyl)picolinic acid

E we shchi) -d o Mn.

According to method 2 using 3-amino-5-fluoro-6-(2-fluorophenyl)picolinate (1.0 equiv.) and PON (5.0 equiv.) 3-amino-5-fluoro-6-(2 -fluorophenyl)picolinic acid with a yield of 90 95. LC/MS - 251.1 (Man), CH-0.80 min.

Synthesis of methyl-3-amino-6-(2,6-difluorophenyl)-5-fluoropicolinate

E E shk i c) -th about Me,

According to method 1 using methyl-3-amino-6-bromo-5-fluoropicolinate (1.0 equiv.) and 2,6-difluorophenylboronic acid (1.3 equiv.) equiv.) 3-amino-6-(2,6-difluorophenyl)-5-fluoropicolinate was obtained with a yield of 94 95. РХ/МСО - 283.0 (MAN), Ch-0.76 min.

Synthesis of 3-amino-6-(-2,6-difluorophenyl)-5-fluoropicolinic acid

E E throw things) | -

OM Nn

According to method 2 using 3-amino-6-(2,6-difluorophenyl)-5-fluoropicolinate (1.0 equiv.) and (OH (1.0 equiv.) -difluorophenyl)-5-fluoropicolinic acid with a yield of 79 95. PH/MO - 269.0 (MH), KE-O, 79 min.

Synthesis of 5-fluoro-6-(2-fluorophenyl)picolinic acid

E in: "ai no -X7-b in)

According to method 1 using b-bromo-5-fluoropicolinic acid (1.0 equiv.) and 2-fluorophenylboronic acid (1.3 equiv.) and Pa(arrOSI--DXM (0.05 equiv.)) 5-fluoro- 6-(2-fluorophenyl)picolinic acid with a yield of 43 95. LC/MS - 236.1 (Ma-H), 0.72 min.

Synthesis of 6-(3,4-difluorophenyl)-5-fluoropicolinic acid

IS

E ki: y shchi) 7 c)

According to method 1 using 6-bromo-5-fluoropicolinic acid (1.0 equiv.) and 3,4-difluorophenylboronic acid (1.3 equiv.) and Pa(arrOSI--DCM (0.05 equiv.)) 6 was obtained -(3,4-difluorophenyl)-5-fluoropicolinic acid with a yield of 70 95. РХ/МСО - 254.1 (MAN), K-0.81 min.

Synthesis of 6-(2,5-difluorophenyl)-5-fluoropicolinic acid

IS

What are we? - c)

According to method 1 using 6-bromo-5-fluoropicolinic acid (1.0 equiv.) and 2,5-difluorophenylboronic acid (1.3 equiv.) and Pa(arrOSI--DCM (0.05 equiv.)) 6 was obtained -(2,5-difluorophenyl)-5-fluoropicolinic acid with a yield of 80 95. РХ/МСО - 254.1 (MAN), K-0.74 min.

Synthesis of 6-(2,4-difluorophenyl)-5-fluoropicolinic acid

IS

And: and but - )

According to method 1 using 6-bromo-5-fluoropicolinic acid (1.0 equiv.) and 2,4-difluorophenylboronic acid (1.3 equiv.) and Pa(arrOSI--DCM (0.05 equiv.)) 6 was obtained -(2,4-difluorophenyl)-5-fluoropicolinic acid with a yield of 79 95. РХ/МСО - 254.1 (MAN), K-O, 75 min.

Synthesis of 5-fluoro-6-(2-fluoro-5-propoxyphenyl)picolinic acid

And where: y shchi) - c)

According to method 1 using b-bromo-5-fluoropicolinic acid (1.0 equiv.) and 2-fluoro-5-propoxyphenylboronic acid (1.5 equiv.) and Pa(arr)SiI2-DCM (0.05 equiv. ) obtained 5-fluoro-6-(2-fluoro-

B-propoxyphenyl) picolinic acid. LC/MS - 294.2 (MN), CI-0.95 min.

Synthesis of 6-(2-fluorophenyl)picolinic acid

IS

MO schi) -b c)

6-(2- fluorophenyl)picolinic acid with a yield of 93 95. LC/MS - 218.0 (Ma-H), Ch-0.66 min.

Synthesis of 6-(2,6-difluorophenyl)picolinic acid

E E

MO schi) - c)

According to method 1 using b-bromopicolinic acid (1.0 equiv.) and 2,6-difluorophenylboronic acid (1.5 equiv.) and Pa(arrOSI»-A-DCM (0.05 equiv.) 6-( 2,6-difluorophenyl)picolinic acid with a yield of 38 95. LC/MS - 236.0 (MH), EC - 0.87 min.

Synthesis of 6-(2-fluoro-5-methoxyphenyl)picolinic acid po,

IS

Most) - c)

According to method 1 using b-bromopicolinic acid (1.0 equiv.) and 2-fluoro-5-methoxyphenylboronic acid (1.3 equiv.) and Pa(arrOSI--DCM (0.15 equiv.)) 6-( 2-fluoro-5-methoxyphenyl)picolinic acid with a yield of 95 95. LC/MS - 248.2 (MAN), CI-0.78 min.

Synthesis of 6-(2-fluoro-5-propoxyphenyl)picolinic acid ra o)

IS

MO schi) - c)

According to method 1 using b-bromopicolinic acid (1.0 equiv.) and 2-fluoro-5-propoxyphenylboronic acid (1.5 equiv.) and Pa(arr)CI--DCM (0.15 equiv.) 6 was obtained -(2-fluoro-5-propoxyphenyl)picolinic acid with a yield of 20 95. LC/MS - 276.0 (MAN), EC-0.87 min.

Synthesis of 6-(2,6-difluoro-4-methoxyphenyl)picolinic acid

E E

Powers) - d c)

According to method 1 using b-bromopicolinic acid (1.0 equiv.) and 2,6b-difluoro-4-methoxyphenylboronic acid (1.3 equiv.) and Pa(arr)CI--DCM (0.15 equiv.) 6-(2,6-difluoro-4-methoxyphenyl)picolinic acid was obtained with a yield of 42 95. LC/MS - 266.1 (MAN), K-0.75 min.

Synthesis of 3-fluoro-6-(2-fluorophenyl)picolinic acid

IS

Mo shchi) ! - c) E

According to method 1 using b-bromo-3-fluoropicolinic acid (1.0 equiv.) and 2-fluorophenylboronic acid (1.5 equiv.) and Pa(arr)CI--DxHM (0.05 equiv.) 3- fluoro-6-(2-fluorophenyl)picolinic acid with a yield of 81 95. LC/MS - 236.1 (Ma-H), 0.72 min.

Synthesis of 3-fluoro-6-(2-fluoro-5-methoxyphenyl)picolinic acid c)

E shchi) - c) E

According to method 1 using b-bromo-3-fluoropicolinic acid (1.0 equiv.) and 2-fluoro-5-

of methoxyphenylboronic acid (1.3 equiv.) and Pa(arrOCl»-DCM (0.15 equiv.) gave 3-fluoro-6-(2-fluoro-

B-methoxyphenyl)picolinic acid with a yield of 89 95. LC/MS - 266.1 (MN), Ch-0.79 min.

Synthesis of 5-fluoro-6-(2-fluoro-5-methoxyphenyl)picolinic acid po,

E i: i shchi) | - c)

According to method 1 using b-bromo-5-fluoropicolinic acid (1.0 equiv.) and 2-fluoro-5-methoxyphenylboronic acid (1.3 equiv.) and Pa(arr)CI--DCM (0.15 equiv. .) obtained 5-fluoro-6-(2-fluoro-

B-methoxyphenyl)picolinic acid with a yield of 86 95. РХ/МСО - 266.1 (МН), Ch-0.79 min.

Synthesis of 6-(4-(benzyloxy)-2-fluorophenyl)-5-fluoropicolinic acid

E ki: y shchi) | - c)

According to method 1 using b-bromo-5-fluoropicolinic acid (1.0 equiv.) and 4-(benzyloxy)-2-fluorophenylboronic acid (1.3 equiv.) and Pa(arr)CI--DCM (0. 15 equiv.) obtained 6-(4-(benzyloxy)-2-fluorophenyl)-5-fluoropicolinic acid with a yield of 28 95. LC/MS - 342.1 (MH), K-1.05 min.

Synthesis of 6-(4-(benzyloxy)-2-fluorophenyl)-3-fluoropicolinic acid

IS

Mo shchi) -dj

About B

According to method 1 using b-bromo-3-fluoropicolinic acid (1.0 equiv.) and 4-(benzyloxy)-2-fluorophenylboronic acid (1.3 equiv.) and Pa(arr)CI--DCM (0, 15 equiv.) obtained 6-(4-(benzyloxy)-2-fluorophenyl)-3-fluoropicolinic acid with a yield of 41 95. LC/MS - 342.1 (MH), K-1.06 min.

Synthesis of 6-(2,6-difluoro-4-methoxyphenyl)-3-fluoropicolinic acid -

E E

Powers) | -dyb c) E

According to method 1 using 6-bromo-3-fluoropicolinic acid (1.0 equiv.) and 2,6-difluoro-4-methoxyphenylboronic acid (1.3 equiv.) and Pa(arr)CI--DCM (0, 15 equiv.) obtained 6-(2,6-difluoro-4-methoxyphenyl)-3-fluoropicolinic acid with a yield of 9 95. РХ/МСО - 284.0 (MAN), Ch-0.74 min.

Synthesis of 6b-cyclohexenyl-5-fluoropicolinic acid i: ai shchi) - c)

6b-cyclohexenyl-5 - fluoropicolinic acid with a yield of 61 95. LC/MS - 222.0 (MH), K-0.52 min.

Methodology Z

Synthesis of 6b-cyclohexyl-5-fluoropicolinic acid where: ay shchi) -db c)

To a degassed solution of b-cyclohexenyl-5-fluoropicolinic acid (1.0 eq.) in MeOH (0.07 M) was added 10 95 Ra/C (0.1 eq.) and the reaction mixture was stirred overnight under a hydrogen atmosphere, which served from a balloon. Then the solution was filtered, washed with MeOH and the filtrate was concentrated to obtain b-cyclohexyl-5-fluoropicolinic acid with a yield of 6595. LC/MS - 224.2 (М--Н), EC-0.95 min.

Synthesis of 5-fluoro-6-(1,4-dioxaspiro|4,5|dec-7-en-8-yl)picolinic acid 04.0 ie: i schi) - c)

According to method 1 using b-bromo-5-fluoropicolinic acid (1.0 equiv.) and 4,4,5,5-tetramethyl-2-(1,4-dioxaspiro|4,5|dec-7-ene- 8-yl)-1,3,2-dioxaborolane (2.0 equiv.) and Pa(arr)CI-DCM (0.2 equiv.) gave 5-fluoro-6-(1,4-dioxaspiro (4, 5|dec-7-en-b8-yl)picolinic acid, LC/MS, 280.2 (MH), -0.66 min.

Synthesis of methyl-5-fluoro-6-(1,4-dioxaspiro(|4,5|dec-7-en-v8-yl)picolinate do) sha c) -j c)

EDH-HCII was added to a solution of 5-fluoro-6-(1,4-dioxaspiro|4,5|dec-7-en-8-yl)picolinic acid (1.0 equiv.) in DCM (0.3 M) (1.0 equiv), DMAP (1.0 equiv), and MeOnH (10 equiv). The reaction mixture was stirred at room temperature for 5 days, then diluted with ethyl acetate, washed with water, brine, dried over magnesium sulfate, filtered and concentrated. The crude product was purified by silica gel column chromatography eluting with 25-50 90 ethyl acetate in hexanes to give methyl-5-fluoro-6-(1,4-dioxaspiroI(4,5|dec-7-en-8-yl )upicolinate as the desired product with a yield of 35 95. LC/MS - 294.2 (MeN), 0.79 min.

Synthesis of methyl-5-fluoro-6-(1,4-dioxaspiro|4,5|decan-8-yl)picolinate od)

E shk: c) and c)

To a degassed solution of methyl 5-fluoro-6-(1,4-dioxaspiroI4,5|dec-7-en-8-yl)picolinate (1.0 equiv.) in

Meon (0.07M) was added at 1095 Pa/C (0.1 equiv.) and the reaction mixture was stirred overnight under a hydrogen atmosphere supplied from a cylinder. Then the solution was filtered, washed with MeOH and the filtrate was concentrated to obtain methyl 5-fluoro-6-(1,4-dioxaspiro|4,5|decan-8-yl)picolinate with a yield of 91 95. LC/MS - 296, 2 (MN), Ch-0.83 min.

Synthesis of methyl-5-fluoro-6-(4-oxocyclohexyl)picolinate

E shk B c) - c)

To a solution of methyl 5-fluoro-6-(1,4-dioxaspiro|4,5|decan-8-yl)picolinate (1.0 equiv.) in acetone and water (1:1, 0.04 M) was added dehydrated oxalic acid (2.0 equiv.) and the reaction mixture was stirred for 3 days. Then the solution was neutralized by adding solid ManHsSoOz, the mixture was added to ethyl acetate and brine, the organic phase was dried over magnesium sulfate, filtered and concentrated.

Methyl 5-fluoro-6-(4-oxocyclohexyl)picolinate was obtained with a yield of 9895. LC/MS, 252.1 (MH), -0.68 min.

Synthesis of methyl-5-fluoro-6-(4-hydroxycyclohexyl)picolinate

N shi c) -- c)

At 0"C, to a solution of methyl-5-fluoro-6-(4-oxocyclohexyl)picolinate (1.0 equiv.) in Mmeon (0.08 M) was added Mavna." The solution was allowed to warm to room temperature overnight, then subjected to distribution between ethyl acetate and brine, the organic phase was dried over magnesium sulfate, filtered and concentrated to give methyl 5-fluoro-6-(4-hydroxycyclohexyl)picolinate as a mixture of two isomers (5:1). LC/MS - 254.2 ( MN), CI-0.63 min.

Synthesis of methyl-6-(4-(tert-butyldimethylsilyloxy)cyclohexyl)-5-fluoropicolinate tv5

E shi Sh c) - c)

To a solution of methyl 5-fluoro-6-(4-hydroxycyclohexyl)picolinate (1.0 equiv.) in DMF (0.15 M) was added imidazole (4.0 equiv.) and TVOM5ZSI (2.5 equiv.). The reaction mixture was stirred at room temperature for 2 days, then added to ethyl acetate, washed with water, brine, dried over magnesium sulfate, filtered and concentrated to give methyl 6-(4-(tert-butyldimethylsilyloxy)cyclohexyl)-5-fluoropicolinate in a yield 97 95 in the form of a mixture of isomers (3:1).

LC/MS - 368.3 (MN), Ch-T1.4 and 1.42 min.

Synthesis of 6-(4-(tert-butyldimethylsilyloxy)cyclohexyl)-5-fluoropicolinic acid

TV5 and: ay shchi) - c)

To a solution of methyl 6-(4-(tert-butyldimethylsilyloxy)cyclohexyl)-5-fluoropicolinate (1.0 equiv.) in tTHF/Meon (2:1, 0.09 M) was added GION (1.5 equiv.) . The reaction mixture was stirred overnight at room temperature, then 1 n. HCl and ethyl acetate, the organic phase was washed with brine, dried over magnesium sulfate, filtered and concentrated to obtain 6-(4-(tert-butyldimethylsilyloxy)cyclohexyl)-5-fluoropicolinic acid in the form of a mixture of isomers (3:1) with a yield of 82 95. LC/MS - 354.2 (MN), EC-1.38 and 1.41 min.

Synthesis of bromo-5-fluoropicolinic acid

M Y shchi) U

To 2-bromo-3-fluoro-6-methylpyridine (1.0 equiv.) in H2O (30 mL) was added potassium permanganate (1.0 equiv.).

The solution was heated at 100 °C for 5 h and then more potassium permanganate (1.0 equiv) was added. After heating for another 48 h, the material was filtered through Celite (4 cm x 2 in) and washed with H2O (150 mL). The combined aqueous solution was acidified with 1N HCl to pH-4, extracted with ethyl acetate (200 ml), washed with MCl, dried over Mazoch, filtered and concentrated to obtain b-bromo-5fluoropicolinic acid (17 95 in the form of a white solid. LCMS (n/2): 221.9 (MN); LC CI-2.05 min.

Methodology 4

Synthesis of 2-(2,6-difluorophenyl)-3-fluoro-6-methylpyridine

Eh hey -

To a solution of 2-bromo-3-fluoro-6-methylpyridine (1.0 equiv.) in THF and water (10:11, 0.2 M) was added 2,6-difluorophenylboronic acid (2.0 equiv.) and fluoride potassium (3.3 equiv.). The reaction mixture was degassed for 10 min, then Raz(aba)z (0.05 equiv.) was added, followed by tri-tert-butylphosphine (0.1 equiv.). The reaction mixture was stirred at 60 "C for 1 hour, then according to LC/MS all the starting substances were consumed. The reaction mixture was allowed to cool to room temperature, it was partitioned between ethyl acetate and water, the organic phase was dried over sodium sulfate, filtered and concentrated .

The crude substance was diluted in EN to 0.1 M, and 0.5 equiv was added. MaVNa to remove aBa.

The reaction mixture was stirred for 1 hour. at room temperature, then the reaction was quenched with water and concentrated in vacuo to remove ethanol. The product was extracted with ether, washed with brine, the organic matter was dried over sodium sulfate, filtered and concentrated.

The crude substance was loaded into silica gel and purified by column chromatography (I5CO) eluting with a mixture of hexanes and ethyl acetate (0595-1095 ethyl acetate). The pure fractions were combined and concentrated to give 2-(2,6-difluorophenyl)-3-fluoro-6-methylpyridine as a light yellow oil with a yield of 86 95. LC/MS - 224.0 (MAN), Ch-0, 84 min.

Methodology 5

Synthesis of 6-(2,6-difluorophenyl)-5-fluoropicolinic acid

E E with and but - )

To a solution of 2-(2,6-difluorophenyl)-3-fluoro-b-methylpyridine (1.0 equiv.) in water (0.05 M) was added KMpO4 (2.0 equiv.) and the reaction mixture was boiled with reflux overnight. Another 2.0 equiv was added.

KMpOi and stirred under reflux for another 8 hours. The solution was cooled to room temperature, filtered through celite and washed with water. The filtrate was acidified with b.n. NCI to rnH-Z, the white precipitate was filtered. The filtrate was further acidified to pH-1 and filtered again. The filtrate was extracted with ethyl acetate until all the product was removed from the aqueous layer. The organic phase was washed with brine and dried over magnesium sulfate, filtered and concentrated. The residue was dissolved in ethyl acetate, washed with 1 n. Mahon, the aqueous layer was acidified to pH. and the white crystals were filtered off. The products were combined to give 6-(2,6-difluorophenyl)-5-fluoropicolinic acid in a yield of 32 95 as a white solid. RH/MS - 254.0 (MAN), EKCH-O, 71 min.

Synthesis of 6-(2,6-difluorophenyl)-3-fluoro-2-methylpyridine

E E

M 'os -d

IS

To a solution of 6-bromo-3-fluoro-2-methylpyridine (1.0 equiv.) in ethanol and toluene (1:1, 0.2 M) was added 2,6-difluorophenylboronic acid, DIEA (5 equiv.) and RaA (PpH3)" (0.2 equiv.). The reaction mixture was heated in a microwave oven at 120 "С for 30 min. The solution was filtered and washed with ethyl acetate. Volatile substances were removed in vacuo and the crude substance was purified using column chromatography on silica gel eluting with ethyl acetate and hexanes (2.5-20 96 ethyl acetate). After concentration of the pure fractions, 6-(2,6-difluorophenyl)-3-fluoro-2-methylpyridine was isolated with a yield of 88 95. РХ/МСО - 224.1 (Man), Ch-0.87 min.

Synthesis of 6-(2,6-difluorophenyl)-3-fluoropicolinic acid

E E m 'os no - ) E

According to method 5 using 6-(2,6-difluorophenyl)-3-fluoro-2-methylpyridine (1.0 equiv.) and potassium permanganate (6.0 equiv.) 6-(2,6-difluorophenyl)- 3Z-fluoropicolinic acid with yield

ZO to РХ/МСО - 254.1 (MAN), KI-O, 70 min.

Synthesis of 2-(2,6-difluoro-3-methoxyphenyl)-3-fluoro-b-methylpyridine

F)

E E where: and -

2-(2,6-difluoro -3-methoxyphenyl)-3-fluoro-6-methylpyridine with a yield of 60 95. РХ/МСО - 254.1 (Ma-Н), K-0.85 min.

Synthesis of 6-(2,6-difluoro-3-methoxyphenyl)-5-fluoropicolinic acid

F)

E E i: ay shchi) | -d c)

6-(2,6 -difluoro-3-methoxyphenyl)-5-fluoropicolinic acid with a yield of 27 95. LC/MS - 284.1 (MH), EC-0.75 min.

Synthesis of 3-fluoro-6b-methyl-2-(2,3,5-trifluorophenyl)pyridine

E E

Well, give it up

To a solution of 2-bromo-3-fluoro-b6-methylpyridine (1.0 equiv.) in dioxane (0.2 M) was added 2,3,5-trifluorophenylboronic acid and Pa(arr)CI--DCM (0.1 eq.). An aqueous solution of sodium carbonate (2M solution, 2.0 equiv.) was added and the reaction mixture was heated in a microwave oven at 120 °C for 15 min. The solution was partitioned between ethyl acetate and saturated Manso's solution, the organic phase was washed with brine, dried over sulfate magnesium, filtered and concentrated. The crude substance was purified by silica gel column chromatography eluting with ethyl acetate and hexanes (1:3) to give 3-fluoro-6b-methyl-2-(2,3,5-trifluorophenyl)pyridine in a yield of 87 95. LC/MS - 242.1 (MH), EC-0.98 min.

Synthesis of 5-fluoro-6-(2,3,5-trifluorophenyl)picolinic acid

E E

E i: i shchi) - c)

Potassium permanganate (10 equiv.) and the solution was heated at 90 "C for 5 h. Upon cooling to room temperature, the solution was filtered and the filtrate was concentrated under reduced pressure to give 5-fluoro-6-(2,3,5-trifluorophenyl)picolinic acid with a yield of 89 95. PH/MS - 272.0 (MH),

E-0.80 min.

Synthesis of methyl-6-bromo-5-fluoropicolinate

And you: "Hey, - )

To a solution of 6b-bromo-5-fluoropicolinic acid (1.0 equiv.) in methanol (0.2 M) was added No5Ox (42 equiv.) and the reaction mixture was stirred at room temperature for 2 h. After completion of the reaction according to LC/MS, the reaction mixture was diluted with ethyl acetate and the reaction was slowly quenched with a saturated aqueous solution of ManHsSoOz. The reaction mixture was poured into a separatory funnel and extracted with ethyl acetate. The organic phase was dried over magnesium sulfate, filtered and concentrated in vacuo to give methyl 6-bromo-5-fluoropicolinate as a white solid (299 95). LC/MS - 233.9/235.9 (MH), Ch-0.69 min.

Methodology 6

Synthesis of methyl-6-(3-(benzyloxy)-2,6-difluorophenyl)-5-fluoropicolinate by

E E i ro) -d c)

To a solution of methyl-6-bromo-5-fluoropicolinate (1.0 equiv.) in THF and water (10:1.01 M) was added 3-(benzyloxy)-2,6-difluorophenylboronic acid (2.5 equiv.) and potassium fluoride (3.3 eq.). The reaction mixture was degassed with nitrogen, then Raz(aba)z (0.25 equiv.) and tri-tert-butylphosphine (0.5 equiv.) were added and the reaction mixture was heated at 80 °C for 1 h. Analysis by LC/MS indicated complete conversion of the starting material to product. The reaction mixture was cooled to room temperature, then concentrated in vacuo and loaded onto silica gel. The crude product was purified by IBESO flash chromatography eluting with ethyl acetate and hexanes (from 0 to 30 95 ethyl acetate) to give methyl-6-(3-(benzyloxy)-2,6-difluorophenyl)-5-fluoropicolinate as the desired product in the form of a light yellow oil with a yield of 96 95. РХ/МСО - 374.0 (Me-Н), K- 1.07 min.

Synthesis of methyl-6-(3-(benzyloxy)-2,6-difluorophenyl)picolinate o.

E E

MO Rio, - c)

By method b using methyl-b-bromopicolinate (1.0 equiv.) and 3-(benzyloxy)-2,6-difluorophenylboronic acid (2.5 equiv.) methyl-6-(3-(benzyloxy)-2 ,6-difluorophenyl)picolinate in the form of a light yellow solid with a yield of 95 95. LC/MS - 356.2 (MH),

MKE1.03 min.

Synthesis of methyl-6-(2,6-difluoro-4-methoxyphenyl)-5-fluoropicolinate -

E E i: i c)

By method b using methyl-b-bromopicolinate (1.0 equiv.) and 2,6-difluoro-4-methoxyphenylboronic acid (2.5 equiv.) methyl-6-(2,6-difluoro-4-methoxyphenyl) was obtained )-5-fluoropicolinate in the form of a white solid with a yield of 85 95. LC/MS - 298.0 (MH), RI-0.89 min.

Synthesis of 6-(2,6-difluoro-4-methoxyphenyl)-5-fluoropicolinic acid -

E E ki: i shchi) -b c)

To a solution of methyl 6-(2,6-difluoro-4-methoxyphenyl)-5-fluoropicolinate (1.0 equiv.) in THF/MeOnH (2:11, 0.09 M) was added GION (1.5 equiv. ) and the reaction mixture was stirred at room temperature for 1 h. The reaction was stopped with 1 n. NS, extracted with ethyl acetate, washed with brine, dried over sodium sulfate, filtered and concentrated to obtain 6-(2,6-difluoro-4-methoxyphenyl)-5-fluoropicolinic acid with a yield of 84 95. LC/MS - 284.1 (MH ), CI-0.76 min.

Methodology 7

Synthesis of methyl 6-(2,6-difluoro-3-hydroxyphenyl)-5-fluoropicolinate

Finn!

E E

S ro, - c)

To a solution of methyl 6-(3-(benzyloxy)-2,6-difluorophenyl)-5-fluoropicolinate (1.0 eq.) in methanol (0.1

M) was added to 1095 Ra/C (0.1 equiv.) in ethyl acetate. The reaction mixture was placed in a hydrogen atmosphere and stirred for 2 hours. After completion of the reaction, the solution was filtered through a layer of celite, the layer was washed with methanol, the filtrate was concentrated in vacuo and methyl 6-(2,6-difluoro-3-hydroxyphenyl)-5-fluoropicolinate was obtained as a gray oil with a yield of 86 95. LC/MS - 284.0 (MN), Ch-0.90 min.

Synthesis of methyl-6-(2,6-difluoro-3-hydroxyphenyl)picolinate

E E

MO yes, - c)

According to method 7, using methyl 6-(3-(benzyloxy)-2,6-difluorophenyl)picolinate (1.0 equiv.), methyl 6-(2,6-difluoro-3-hydroxyphenyl)picolinate was obtained in the form of - brown solid with a yield of 96 95. LC/MS - 266.0 (MAN), EC-0.68 min.

Synthesis of methyl 6-(2-fluoro-5-formylphenyl)picolinate

N

IS

And: ro, - c)

To a solution of methyl β6-bromopicolinate (1.0 eq.) in DME (0.03 M) in a microwave oven vessel was added Pa(arry)Si--DCM (0.05 eq.), 2-fluoro-5- formylphenylboronic acid (1.5 equiv.) and 2M MazbOz (2 equiv.). The reagents were heated at 120 "C for 20 min. A mixture of the desired product and the corresponding carboxylic acid was detected by LC/MS, the reaction mixture was diluted with ethyl acetate, washed with HCl (pH.-5), the acidic phase was extracted with ethyl acetate, the organic layers were combined dried over magnesium sulfate, filtered, and concentrated in vacuo to give a light brown solid. The solid was dissolved in Meon and treated at room temperature with 3 equiv TMCO-diazomethane. After complete conversion of the carboxylic acid to the corresponding methyl ether, the reaction mixture was concentrated to vacuum and the crude substance was purified by column chromatography on silica gel (LCSO) eluting with 30 95 ethyl acetate in hexanes to give methyl 6-(2-fluoro-5-formylphenyl)picolinate as a yellow solid with a yield of 58 95. /MS - 260.0 (MAN), 0.70 min.

Synthesis of (E)-methyl-6-(2-fluoro-5-(prop-1-enylphenyl)picolinate

J

IS

M's d-O -d c)

To a solution of methyl 6-(2-fluoro-5-formylphenyl)picolinate (1.0 equiv.) in MeOH (0.17 M) was added ethyltriphenylphosphonium bromide (1.0 equiv.), then sodium methoxide (1.5 equiv. ). The reaction mixture was heated at 65 °C for 5 hours, then cooled to room temperature and concentrated in vacuo.

The crude substance was purified by column chromatography on silica gel (IZSO) eluting with 50% ethyl acetate in hexanes and (E)-methyl-6-(2-fluoro-5-(prop-1-enyl)phenyl)picolinate was obtained as white solid with a yield of 81 95. LC/MS - 272.0 (MH), -0.73

MIN.

Synthesis of methyl 6-(2-fluoro-5-propylphenyl)picolinate

IS

M osh

She c)

1095 Ra/С (0 .5 equiv.) and the reaction mixture was placed in a hydrogen atmosphere and stirred overnight. The mixture was filtered through a celite pad and washed with Meon. The filtrate was concentrated in vacuo and methyl 6-(2-fluoro-5-propylphenyl)picolinate was obtained as a light gray oil with a yield of 97 95. LC/MS - 274.2 (MH), K-0.61 min.

Synthesis of 6-(2-fluoro-5-propylphenyl)picolinic acid

IS

She: but -d )

To a solution of methyl 6-(2-fluoro-5-propylphenyl)picolinate (1.0 eq.) in THF was added lithium hydroxide (10 eq.) and the reaction mixture was stirred at room temperature for 1 h. THF was removed in vacuo and the remaining alkaline phase was acidified with concentrated HCl. The aqueous layer was extracted with ethyl acetate (2x), the organic phase was dried over sodium sulfate, filtered and concentrated to obtain 6-(2-fluoro-5-propylphenyl)picolinic acid with a yield of 35 95. РХ/MO - 260.2 (МН), КЕ -0.36 min.

Methodology 8

Synthesis of methyl 6-(2,6-difluoro-3-"(trifluoromethylsulfonyloxy)phenyl)-5-fluoropicolinate

OT

E E

«y ro) -d c)

To a solution of methyl 6-(2,6-difluoro-3-hydroxyphenyl)-5-fluoropicolinate (1.0 equiv) in DCM (0.2 M) was added DIEA (2.0 equiv) and 1.1, 1-trifluoro-M-phenyl-M-(trifluoromethylsulfonyl)methanesulfonamide (1.5 eq.). The reaction mixture was stirred overnight at room temperature. The reaction was quenched with water, the organic phase was dried over sodium sulfate and concentrated. The crude substance was purified by HSO chromatography eluting with ethyl acetate and hexanes (0-30 95 ethyl acetate).

Pure fractions were concentrated and methyl-6-(2,6-difluoro-3-

(trifluoromethylsulfonyloxy)phenyl)-5-fluoropicolinate as the desired product in the form of a clear oil with a yield of 68 95. LC/MS - 416.1 (MAN), Ch-1.08 min.

Synthesis of methyl 6-(2,6-difluoro-3-"(trifluoromethylsulfonyloxy)-phenyl)picolinate

OT

E E

Mo ro, - c)

According to method 8, using methyl 6-(2,6-difluoro-3-trifluoromethylsulfonyloxy)phenyl)-5-fluoropicolinate (1.0 equiv.), methyl 6-(2,6-difluoro-3-(trifluoromethylsulfonyloxy) was obtained )phenyl)picolinate in the form of a colorless oil with a yield of 299 95. RH/MSO - 397.9 (MAN), Ch-1.03 min.

Synthesis of 6-(2,6-difluoro-3-methylphenyl)-5-fluoropicolinic acid

E E

Su no t c)

Methylboronic acid (3 .0 equiv.) and potassium carbonate (3.0 equiv.). The reaction mixture was degassed with nitrogen for 10 min, then Pa(PpNH3)a (0.1 equiv.) was added to the solution and heated at 100 °C for 3 h. At this point, LC/MS of the reaction mixture indicated complete conversion to carboxylic acid ( MA-Н-268). Cooled to room temperature and water and ethyl acetate were added. These two layers were separated, the aqueous phase was acidified with concentrated HCl to pH and extracted with ethyl acetate. The organic phase was dried over sodium sulfate, filtered and concentrated in vacuo to give 6 -(2,6-difluoro-3-methylphenyl)-5-fluoropicolinic acid in the form of a clear oil with a yield of 97 95. LC/MS - 268.1 (MeN), -0.82 min.

Synthesis of methyl-6-(2,6-difluoro-3-methylphenyl)picolinate

E E

Mos rio, -d c)

To a solution of methyl 6-(2,6-difluoro-3-"-"trifluoromethylsulfonyloxy)-phenyl)picolinate (1.0 equiv.) in toluene was added Ra(arrOCI--DCM (0.1 equiv.), then dimethylzinc (3.0 equiv). The solution turned from orange to bright yellow. The reaction mixture was heated at 80 °C for 2 h and then LC/MS analysis indicated complete conversion to the product. The reaction mixture was cooled to room temperature, diluted with ethyl acetate and washed with brine. The organic layer was dried over magnesium sulfate, filtered and concentrated in vacuo to give methyl 6-(2,6-difluoro-3-methylphenyl)picolinate as a brown oil in quantitative yield. LC/MS - 264, 0 (MAN), K-0.90 min.

Synthesis of 6-(2,6-difluoro-3-methylphenyl)picolinic acid

E E

MO shchi) ! -dj c)

To a solution of methyl 6-(2,6-difluoro-3-methylphenyl)picolinate (1.0 equiv.) in THF was added sodium hydroxide (10 equiv.) and the reaction mixture was stirred for 2 h. The solution was diluted with ethyl acetate and washed with 1 n. Mahon (2x). Aqueous alkaline washing solutions were combined and acidified with concentrated NSI. The acidic aqueous phase was extracted with ethyl acetate (2x), the combined organic layers were dried over magnesium sulfate, filtered and concentrated in vacuo to give 6-(2,6-difluoro-33-methylphenyl)picolinic acid as a white solid with a yield of 85 95 LC/MS - 250.0 (MH), KE-O, 76 min.

Synthesis of 6-(3-ethyl-2,6-difluorophenyl)picolinic acid

E E

MO shchi) -7b c)

To a solution of methyl 6-(2,6-difluoro-3-"-"trifluoromethylsulfonyloxy)-phenyl)picolinate (1.0 equiv.) in toluene (0.15 M) was added Ra(arr)CI.--DCM ( 0.1 eq.), then diethylzinc (3.0 eq.). The solution turned from orange to bright yellow. The reaction mixture was heated at 70 "C for 2 h, and then LC/MS analysis showed the presence of a 1:3:1 mixture of the hydrolyzed product, the desired product, and an unknown byproduct. The reaction mixture was cooled to room temperature, diluted with ethyl acetate, and washed with with 1N Mahon (2x) The organic layer was dried over magnesium sulfate, filtered and concentrated in vacuo to give a brown oil.

The oil was redissolved in THF and treated with 1 n. Mahon within 1 hour. Then the reaction mixture was diluted with ethyl acetate and washed with 1 n. Mahon (2x). Alkaline washing solutions were combined, acidified with the help of concentrated NS! and extracted with ethyl acetate (III).

The organic phase was dried over magnesium sulfate, filtered and concentrated in vacuo to give 6-(3-ethyl-2,6b-difluorophenyl)picolinic acid in the form of a light brown oil with a yield of 299 95. LC/MS - 264.1 (Ma- H), Ch-0.88 min.

Methodology 9

A homogeneous solution of 1 equiv. of each amine, carboxylic acid, NOAT and EDH in DMF at a concentration equal to 0.5 M was kept for 24 h. and then water and ethyl acetate were added. The organic phase was dried over sodium sulfate and purified by silica gel column chromatography eluting with ethyl acetate and hexanes to give the desired protected amide. Alternatively, the crude reaction mixture was immediately purified by HPLC. After lyophilization, a salt of the protected amide was obtained from TFC. Alternatively, HPLC-derived fractions could be added to

EtOAc and solid Mg2CO3, separate and wash with MCl(a). After drying over Mdzoha, filtration and removal of volatile substances in a vacuum, the protected amide was obtained in the form of a free base. Alternatively, the crude reaction mixture was used in the step of removing the protective group without further purification.

If an amine protected by M-Vos was present, it was removed by treatment with an excess of 4M HCl/dioxane mixture for 14 h. or by treatment with 25,956 TFC/SNeoSi" for 2 hours. After removal of volatile substances in a vacuum, the substance was purified by ZF HPLC and after lyophilization, the amide was obtained in the form of a TFC salt. Alternatively, the HPLC fractions could be added to EIOAc and solid MgCO3, separated and washed with Mass).

After drying over Ma5O5, filtering and removal of volatile substances in a vacuum, the free base was obtained. After dissolving in a mixture of MeSM/H2HO, adding 1 equiv. 1 n. NOI and lyophilization obtained the amide salt from NOI.

If M-ros-1,2-aminohydroxycyclic carbamate was present, before removal of the Bos protecting group, the cyclic carbamate could be cleaved by treatment with С52СО3 (0.5 equiv.) in ethanol at a concentration equal to 0.1 M for 3 h . After removal of volatiles in vacuo, the Bos amino protecting group was removed as described above.

If the M-Boc, OAc group was present, the acetate group could be cleaved by treatment with KgCO3 (2.0 equiv.) in ethanol at a concentration equal to 0.1 M for 24 h before removing the Boc protecting group.

If the M-phthalimide group was contained, then the protective group of the amino group was removed by treatment with hydrazine in MeOH at 65 "С for 3 h. After cooling and filtering off the white precipitate, the filtrate was concentrated and purified by ZF HPLC and the amino amide was obtained.

If the TVOM5 ether was contained, the protecting group was removed before removing the Bos protecting group by treatment with a mixture of 6 n. NSI, THF, methanol (1:2:1) at room temperature for 12 hours. After removal of volatiles in vacuo, the Bos amino protecting group was removed as described above. Alternatively, the TVOM5 ether and the Bos group could be removed using a mixture of 6 n. NOAH,

THF, methanol (1:2:1) by holding at RT for 24 hours. or heating at 60 C for 3 hours.

If an OMe group was present, this protecting group was removed by treatment with 1 M VVhz in DCM (2.0 eq.) for 24 h. Water was added dropwise and the volatiles were removed in vacuo.

The substance was purified by reverse-phase HPLC as described above.

If an OVp group was present, this protecting group was removed by treatment with 10 95

Ra/C (0.2 equiv.) in a hydrogen atmosphere in ethyl acetate and methanol (1:2). After completion of the reaction, the reaction mixture was filtered through celite, washed with methanol, and the filtrate was concentrated in vacuo.

Synthesis of (Z-)-3-amino-N-(4-(3-amino-4-hydroxycyclohex-1-enyl)pyridin-3-yl)-6-(2,6-difluorophenyl)picolinamide

N

Nam E E and no zi M Kh

M c) MN"

According to method 9 (17-)-tert-butyl-3-(33-aminopyridin-4-yl)-6-(tert-butyldimethylsilyloxy)cyclohex-2-enylcarbamate and 3-amino-6-(2,6-difluorophenyl) picolinic acid was introduced into the combination reaction and the protective group was removed to obtain (/-)-3-amino-M-(4-(3-amino-4-hydroxycyclohex-1-enyl)pyridine-

3-yl)-6-(2,6-difluorophenyl)picolinamide as a salt with TFC. RHMS (t/l2): 438.2 (MNU), RH Ke2.00 min.

Synthesis of (1u/-)-3-amino-N-(4-(3-amino-4-hydroxycyclohexyl)-pyridin-3-yl)-6-(2,6-difluorophenyl)picolinamide

N

TO US

? E E and no - c) MN

M 2

According to method 9 (17-)-tert-butyl-5-(3-aminopyridin-4-yl)-2-(tert-butyldimethylsilyloxy)cyclohexylcarbamate and /3-amino-6-(2,6-difluorophenyl)picolinic acid were introduced in the combination reaction and the protective group was removed and (/-)-3-amino-N-(4-(3-amino-4-hydroxycyclohexyl)pyridin-3-yl)-6-(2,6-difluorophenyl)picolinamide was obtained in in the form of a salt from TFC with a yield of 18 95. LCMS (t/2): 440.3 (MNU), LC h-2.04 min.

According to method 9, the following compounds were obtained:

Table 1

Example | Structure of RH/MS RHMS Chemical name Y

SHU (MAN at | (KG at

ORSI) PRSI)

What 7 Y Zhiralnyi! 6-(2,6-difluorophenyl)-5-no CH E and ! fluoro-M-(4-41K,35,55)-3-slya!

And | iii 442.2 0.75 hydroxy-5- i Zhe : i | mo! methylcyclohexyl pyride mn-3-ylncolinamide and OH Khvvlnii M-(4-(3K4K,553-3- and not in ! amino-4-hydroxy-5-ki ku VI | methylpiperidin-1-2 С 5 474, 3 0.52 and ilopridin-z-yl)-6-(2,6- difluoro-4-! | hydroxyphenyl)-5- and ! ! ' fluoropicolinamide

ШИ Son puchirate now Z-amino-M-(4-mn, not Y is E (KAK, 55)-3-amino-4- ! i o m 473.3 0.55 methylpiperidin-1-yl iridin-3-yl)-6-(2,6-difluorophenyl)-5-fluoropicolinamide

OH Chralnyi M-(44(В3К4К,553-3-bu | : amino-4-hydroxy-5- : «v nut | methylcyclohexyl) 4 s bu 473.3 0.52 I shk

C 4 pyridin-3-yl)-6-(2,6-difluoro-4-dihydroxyphenyl)-5-! --

Example )| Structure РХ/MS РХ/MS Chemical name

Co. (MAN at | (KI at

ORSI) ORSI) on Occhiral M-(4-(18,3K,48,55)-3-er amino-4-hydroxy-5-

And tone! methylcyclohexyl)

I I I ts

According to M. Sho

C o 455.3 0.55 pyridine-3-1l)-5-fluoro-b-(2-fluoro-5-hydroxyphenyl) picolinamide

Н Chiral M-(4-(18K,35)-3- sia nm i i in aminocyclohexyl) and mit) 407.2 0.52 pyridin-3-yl)-6-(2-fluoro- what ! I r o 4-hydroxyphenyl)

I M I picolinamide

Shchi no Chralnyi M-A(4-(C18,35)-3- n. - ! she KE aminocyclohexyl) 7 "and 407,2 0,53 pyridin-3-yl)-6-(2-fluoro-

She a I mo 5-hydroxyphenyl) picolinamide

HO Chiral M-A4-(1K,35)-3- nn E v aminocyclohexyl) m pyridin-3-yl)-6-(2,6-8 x A. A 425.2 0.54 and mech

And about difluoro-4-hydroxyphenyl) picolinamide

I OH o Chiralmium M-(44(1K,35)-3-nm E aminocyclohexyl)

E a "what" pyridin-3-yl)-5-fluoro-o- (9 x i i 425.2 0.53 i y 5-rtor sh in (2-fluoro-4-hydroxyphenyl)! picolinamide

"Example | Structure What RHLMS GRH/MS Chemical name

Ho | (MAN at | (VE at and IZ

SHRSI) | ORSI)

ON Hralnii | SHO! OST MAV) 5self-

Жеру" І і 3, З-dimethylacyclo- or г І ! hexyl)niridin-З-ія)-5-. gen 453.3 and 0.58 and і

Her. swarm 3 | and fluoro-b-(2-fluoro-4-

M and hydroxyphenyl) ! | picolinamide n with CH ukral | CM444(1K,5K)-5-amino-

SNU many! nefiche she shk: | and 3,3-dimethylcyclo- in I and I | zpi n-3-yl)-6 : hexyl)pyridin-3-yl)-6-

VI and a and 459.2 0.7 ' | mo (2,6-difluoro-3- methylphenyl)-5- i i, ! and fluoropicolinamide

Shchi SHY shk SNujchiral SHHI I M44-(1K.5K)-5-amino-sn MH H "in lk: 3,3-dimethylcyclo-

EC I | Y I hexyl)pyridin-3-yl)-o» 12 ! and zl 451.1 0.65 Sho and 8 Sh (2,6-difluoro-3-methylphenyl) and ! and oh! and picolinamide tt pi o iii in li pet tu on with "ON Khralnyi and MA4-4(3K4K,55)-3-

Mn ne t, E y y t " ve ak: amino-4-hydroxy-5-in LY methylpiperidine-1- 13 | Z Z 454.2 0.58 shi sche ' | mo il)niridin-3-yl ) -6-(2,6- !difluoro-3-methylphenyl) ! | | picolinamide ru shi MA K,353-3- and Mn, and | and

S aka! aminocyclohexyl) ! and | and 14th grade | 445.2 0.62 pyridin-3-yl)-5-fluoro-6-the!

SYA i | (2,3,5-trifluorophenyl) picolinamide

SHCI on she: x sn ratnyy What MAV K4K,58)-3- and No, "MN, der H - - and is: amino-4-hydroken-Z-

And і or І | in c15: pike and 457.5 0.63 and methylniclohexyl)

PI about pyridin-3-yl)-6-(3-ethyl-

AI what | 2,6-difluorophenyl)

Example | Structure.G VRH/MS OO RH/MS Chemical name) 00000. and (MEN at |(KGat

ORSI) MRS picolinamide I t tires. and M-(4-4384K,55)-3 - pc M | | amino-4-hydroxy-5- 16 EE and methylpiperidine-i-

M MI 428.2 0.03 xyl)pyridine-Z-yl)-6-

Zak M se o cyclohexyl-5- -

M fluoropicolinamide

SHY it about fbrtsnivanny | SHY M-(4-(1K3845,55)-3- ne, k Kn; | En I E | . and IF se amino-d-hydroxy-5-m. 2 V | methylcyclohexyl) (17 in: with 4673 0 ,64 z o pyridine-3-yl)-6-4(3-ethyl-2.6-difluorophenyl)picolinamide in the form of kh.oMH, ІІ, І

FEE E amino-4-hydroxycyclo- - in . , yin | hexyl)niridin-Z-1l)-6 ne: С ха 453.2 0.66 Sh zo (Z-ethyl-2,0-difluorophenyl) picolinamide me e z | aminocyclohexyl) 19 u. 437.2 0.67 pyridin-3-yl)-6-(3-ethnyl- 5 2,6-difluorophenyl) picolinamide

ИМЖБ-Ш2Ш25 dreams are still not E | - EE E . aminocyclohexyl

Av

I and | 441.2 0.64 : pyridin-3-yl)-6-(2,6-

Well, whose in! difluoro-3-methylphenyl)-5-fluoropicolinamide

Example | Structure and РХ/МСО РХХ/МС Chemical name

Me (MaN at | (BE at

ALL SHRSI) - . poison g snkalnia | M-(4-(1К,35,55)-3- ; "MN, ! . that | | amino-5- - m | !

Ash | ! methylcyclohexyl) ! 21 bu 4512 0.7 s Ge | pyridine-3-4ya)-6-(3-ethyl- and 2,6-difluorophenyl) picolinamidon Z CH kraleny Mav, 55) osu and | amino-4-hydroxy-5-dn ! vn I | methylpiperidine-i- 22 zbu 472.3 0.59 i. ! me | yl)pyridin-3-yl)-6-(2,6-difluoro-3Z-methylphenyl)»5Z-fluoropicolinamide tk sht GLUE ZUZ er and amino-4-hydroxy-5- 23 ie n i i i i 429.3 0.53 methylpiperidine -i1- p

Sulyl)pyridine-3-1l)-6-(2-fluorophenyl)picoline-amidone Mralnyi MAZKA, 5)3-

We have a dream where .

St E amino-4-hydroxy-3-m MU | and n | methylpiperidine-1-24 y 458.3 0.51 shk sho z IY veyl)pyridin-3-yl)-6-(2,6- difluorophenyl)-3- and I fluoropicolinamide o 25 ral | M-(4-(3K.4B,55)-3-ru and | amino-4-hydroxy-5-

M goals! . n her | methylpiperidine-1-aka 440.3 0.54 2, ,

FI o yl)pyridin-3-1l)-5-fluoro-6-2-fluorophenyl)picolinamido n g Mralny M-(4-(38,48,55)-3- peru As: | amino-4-hydroxy -5- 26 m MU | 440.2 0.53 and bu SHHI | , , methylpiperidin-1- schi y | ip)pyridin-3-yl)-6-(2,6-

Example ) Structure LC/MS 0 LC/MS Chemical name What

M (MAN at | (KE at '

SHRSI) all pt CH oral M-(4-(18,35,55)-3- pschi and amino-95-ra: "Zhshi methylzniclohexyl) 27 zaru 455.3 0.06 y , mo pyridine-3- yl)-6-(2,6-difluoro-3-methylphenyl)»-5-fluoropicolinamide (aminocyclohexyl) 28 n 423.2 0.61 pyridin-3-yl)-6-(2,6-

ECO OM: s / o difluoro-3-methylphenyl) picolinamide

SI St uchiral 1 m-4-(18,35,55)-3- ether amino-5-tone methylniclohexyl) 29 data My t 437,2 0,64 | AND

FI pyridine-3-1l)-6-(2,6- difluoro-3-methylphenyl) picolinamidon S Y M-(4-(1 КК, АК,55)-3-mncho or amino-4-hydroxy- 5- tn HER methylcyclohexyl) scan y 453.2 0.59 , I si / in pyridin-3-yl)-6-42,6-difluoro-3-methylphenyl) picolinamide "on SNchiral MAYA18.3К45.55)- 3-por E amino-4-hydroxy-5-thn methylylylclohexyl) 3 С se 453.2 0.6 y , z | o pyridin-3-yl)-6-(2,6- difluoro-3-methiaphenyl) picolinamide

Example / Structure of GRH/MS RH/MS | Chemical name of m (MeNpri (VHpri

ORSI) force

STVA Khrelny | | MA44(1E,35)-3- nd is in | | aminocyclohexyl) to I, which m | pyridine-3-1l)-6-(2,65- 32 or 443,2 0,55 | P 2 5 | difluoro-4-hydroxyphenia)-5- fluoropicolinamido Khualniai M-(44(1K,35)- 3- nm c ШЭ 2 E | aminocyclohexyl)

IS

"shi | | pyridin-3-yl)-5-fluoro-6- 33 x B 425.2 0.53 fluoro mo | (2-fluoro-5- hydroxyphenyl) picolinamido no Kan ГМ-(4-((K, 35)-3-n | I. ! i OT | aminocyclohexyl)

CHU | | - m n Y | | tshridin-3-yl)-3-fluoro-6- 34 ho Sh 425.2 0.52 Shchi P fluorine

Like | | (2-fluoro-5- : hydroxyphenyl) picolinamide

GT neoslk hralnia | ny M-(4-(18,35,55)-3- in sn, E | | amino-5-ny | | methylcyclohexyl) denta 4 0.56 SHHI I - / 5 | | tyridin-3-yl)-6-(2-fluoro-! | 5-hydroxyphenyl) and | picolinamide on amino-5-no methylcyclohexyl)

As M. same with6 that. a 439.2 0.57 pyridin-3-yl)-6-(2,6-difluoro-4-hydroxyphenyl): picolinamide

Example Structure of PT РХМС РХ/MS Chemical name sh

M (MAN at | (KE at !

BRSI) AXLE)

OH xChiral M-(4-(41К,35,55)-3-nal sn, СХ, amino-5-m methylcyclohexyl) 37 i A, 4 0.56 Shchi shchi 5 pyridin-3-yl)-6-( 2-fluoro-4-hydroxyphenyl) picolinamide

GT UN Khirvlenny SHY 1 M-(44(1838,55)-3-

Ve n, sn, o amino-3-methylcyclohexyl) some M Zone 38 zni r. 439.2 0.57 pyridin-3-yl)-5-fluoro-6-(2-fluoro-4-hydroxyphenyl) picolinamide and

HO fatty M-(4-41K,35,55)-3-nm sn, E amino-5-no methylcyclohexyl)

M vu i 39 4 Y 5 k 439.2 0.55 pyridin-3-yl)-3-fluoro-b- (2-fluoro-4- hydroxyphenyl) picolinamidon "EN phkralny Sho TMA440K3B,45,55)33-

Nesy ie MN; IS - .

SE amino-4-hydroxy-5-vn" and methylcyclohexyl)

Su y: 471.2 0.62 y shi that pyridine-3-1l)-6-4(2,6- I difluoro-3-methiaphenyl)-5-fluoropicolinamidon SNeouralny I M-44(1K,3K,453 -3- chem, D. s E E amino-4- tn "its hydroxycyclohexyl) 41 7 сх 439.2 0.57 ! zu K 5 pyridine-3-yl)-6-(2,6-difluoro-3- methylphenyl) picolinamide

(Example |Etructure 0000000 РЖ/MS IRЖЖ/MS -|Chemical name of them (MN at | (KG at

ORSIZ ORSIA

SHY Zhvalny 00010000 | TM K35,55)-3-amino-5-methylcyclohexyl) 42 vo: 4572 0.6 pyridin-3-yl)-6-(2,6-difluoro-4-hydroxyphenyl)-3-fluoropticolyzamide and and shi NO. o Khralnii ny OTMYA4401K35,585)3-. and: that amino-3-rich methylcyclohexyl) 43 and o 439.2 0.56 pyridin-3-yl)-3-fluoro-6-(2-fluoro-5-hydroxyphenyl)pticolinamidon Studvalnyi MAK E4K,55)- 3- in; and in ac: amino-4-hydroxyn-5-re methylcyclohexyl) 44 ai -4 471.2 0.62 ! - more about | pyridine-3-17l)-6-(2,6-'difluoro-3-methylphenyl)-5-fluoropicolinamide CH, E | amino-5-rya and Her! 4531 0.v1 methylcnclohexyl)pyryl schi A o in-3-yl)-5-fluoro-6-(2- fluoro-5-methoxyphenyl) picolinamido g Chiralene M-a-(K35,55)-3-b, sn, E amino-5-16 and | b and zvi 0.56 methylcyclohexyl)pyride still o | nn-3-yl)-5-fluoro-6-(2-fluoro-5-hydroxyphenyl) picolinamide

Example | Structure РХ/MS РХ/MS Chemical name

Me (MeN at (KI at

ORSI) RSI) in od bo sciateny Mak KYAK,55)-3- no. what i i i s i: i amino-4-hydroxy-5- ; t MU

Su sh | methylcyclohexyl) 47 mo 487.1 pyridine-3-yl)-6-(2,6-difluoro-3-methoxyphenyl)-5-fluoropicolinamide in: Bra | | M-(4-(41K,35)-3-nom 54 aminocyclohexyl)

Grove in Zhe E

A. vya 4272 0.59 pyridin-3-yl)-6-(2,3,5-

This is I. trifluorophenyl) picolinamide

Hidalny M-4-(1K,5K)-5-amino-one is a 3,3-amino acid.

And nn | dimethylcyclohexyl) 49 Сg ze 455.2 0.63 , and !

I o pyridin-3-yl)-6-(2,6-difluorophenyl)-5-fluoropicolinamide

What is Kirapeny Sh M-(4-401K,38,4K,55)-3-

Mn es I: E amino-4-hydroxyn-5-

AND

EN m) methylcyclohexyl)

ST zr 457.2 0.58 , , mo pyridin-3-yl)-6-(2,6-difluorophenyl)-5-fluoropicolinamide

Chiral that 3-amino-M-(4- on sleep E (18,35,55)-3-amino-5-

And not methylcyclohexyl) 51 and 456.1 0.58

Example | Structure LC/MS LC/MS Chemical name x (MaeNopri | (Vbpri : PYRSII ORSI) n Chiral M-(4-415.35,45,5K)-3- doAya amino-4-hydroxy-3-52 tn methylcyclohexyl) zu i 439.1 0.68 in o pyridin-3-yl)-6-(2,6- difluorophenyl) picolinamide and OH Khfralny that I МА44И1В,353-3-

PC aminocyclohexyl) 53 and 4133 055 pyridin-3-yl)-5-fluoro-6-

C ((18,45)-4-hydroxycyclohexyl) tricholineamide I Chiral | shi M-(4-(1K35,55)-3- I

Nachkit and Sn, | | amino-5- di 5, | | methylcyclohexyl) 54 Fe Z (427.3 | 0.59 pyridin-3-yl)-3-fluoro-6- ((15.45)-4- hydroxycyclohexyl) picolinamide that OH ball t | M-(4-41К,35,55)-3-nm thiamino-5-mzha and methylcnclohexyl) r i: 427.3 0.55 pyridin-3-yl)-5-fluoro-6-y (1 g ,4g)-4-hydroxycyclohexyl) picolinamide

ON Chiraleny | M-(4-4(IK,35)-3- aminodiclohexyl) 56 re KZ Y 4133 48 pyridin-3-yl)-5-fluoro-6- zy 8 ((1g,4g)-4- hydroxycyclohexyl) picolinamide

Example / Structure of РХ/MS РХ/MS Chemical name (ze (MAN at | (KI at

PRSO 0 ORS)

Prchu Zhralnyi and M-(4-(18,35,55)-3-nal that amino-5-57 and | and iz3a 0,4 methylcyclohexyl) or I 6 pyridin-3-yl)-5-fluoro- 6- (2-fluorophenyl) picolinamide Chirapny o M-(44(018,35,553-3- noi sn, | se amino-5- 58 r ii | 419.3 0.67 methylcyclohexyl) ek i ipidin-3 -yl)-6-(2-fluoro-5-methylphenyl) picolinamide not Hypalin M-(4-(1K,35,55)-3- nm sn, E amino-5-zo r ily i i i i i i i i i i i i methylcyclohexyl) and pyridin-3-yl)-5-fluoro-6-(2-fluoro-5-methylphenyl) picolinamide

Eurapny M-(34-(1K,35,55)-3-p sn, E amino-5-p u Y and methylcyclohexyl)

Zh 441.2 0.70 zn / in pyridin-3-yl)-6-(2,3-difluorophenyl)-5-! fluoropicolinamide

E chiralen M-(4-(18,35,55)-3-n; Sn, E amino-5-vy p / | y 44172 0.68 methylcyclohexyl) zm b pyridin-3-yl)-6-(2 ,5-difluorophenyl)-5-fluoropicolinamide

Example Structure ST ВХМС ОО ТРХУМС С Chemical name

Me (MAN at | (KE at

ORSI) ORSI)

Hubix M-(4-CYC,15,55)-3-n, CH amino-5-methylcyclohexyl) 62 pan 405.2 0.67 pyridin-3-yl)-6-(2- fluorophenyl) picolinamide extended M-(4-(1К,35,55)-3-

We had a: amino-5-methylcyclohexyl) 63 sa: 423.2 0.65 | shi zi | o pyridine-Z-yl)-6-(2,0- !hhalin difluorophenyal) picolinamide onoron MA TsIV, K45)-3-

KOM I

FI Ek amino-4-bono and I. What would ha | hydroxycyclohexyl) 64 Ca o 483.2 0.66 | pyridine-3-1l)-5-fluoro-6-

O-fluoro-5-propoxyphenyl) picolinamidon, I you

SE in | | ІІ8,38,45)-3-amino-4-

M g and how | and | hydroxycyclohexynl) 65 | br 458.1 0.55 What ShSH still about nn, | | pyridine-3-yl)-64(2,6-difluorophenyl)-5-fluoropicolinamide one pt 3-amino-M-(4-

I and "MN, I dU- with and F in ((18,38,45)-3-amino-4-

E to и и En | and hydroxycyclohexyl) 6b | C and 440.1 0.55 | I em 5 mn, pyridine-3-yl)-5-fluoro-b" (2-fluorophenyl) picolinamide

Example | Structured 00000000 | РХ/УМС | RH/MS | Chemical name.

Me (MAN pri | (Ki pri

SRS) PROS)

ZD (a B, 38.45)-3-amino-4-in and E hydroxycyclohexyl) 67 FI o Mn, 498.2 0.66 pyridin-3-yl)-5-fluoro-6-(2-fluoro-5- propoxyphenyl) picolinamide ono och and GG 3-amino-M-(4- Shchi

S R (18,3K,453-3-amino-4-

EN 7 | | hydroxycyclohexyl) Щ С В тво 0.5 pyridin-3-yl)-65-(2-fluoro-m o mn, ryd P 5-propoxyphenyl) picolinamide kratin M44(k35,55)-3- (nm sn, E amino -5- 69 и 'sh Й 4414 0.67 methylcyclohexyl) o 0 SE pyridin-3-yl)-6-(2,6- difluorophenyl)-3"- fluoropicolinamide

Chiral M-(4-41K,35,55)-3-amino-5-one (His and methylcyclohexyl) 70, 441.3 0.70 and 43-64,6-s o pyridine-3-1l) -6-(2,6 difluorophenyl)-5-fluoropicolinamide t Melnyk 1 OT M44(8,385K)33 tot | amino-5-

Ash in and | methylcyclohexyl) n C 7 C pyr in-3-yl)-6-(2,6-

M ge shsh rid? difluorophenyl)-3-fluoropicolinamide

Example | Structure РХ/MS РХ/MS Chemical name

Le (MeAN at |((KE at

MORSI) ORSI) oral | TASTE E | amino-5-

I - E and and tv | and methylcyclohexyl) and 72' b B. 441.3 0.70 : s i o pyridine-3-yl)-6-2,6- and difluorophenyl)-5-fluoropicolinamidon 5 Zhvalny 1 M-4-(1K, 3K.45,55)-3-

Mu yin I MM, EE. no!

S E | | amino-4-hydrokei-5- I sa E | N t M N N N

EN | p. and methylcyclohexyl) and 73 au 453, 07 Shchi and ! mo | and pyridin-3-yl)-5-fluoro-be (2-fluoro-5-methylphenyl) and | ! and picolinamide ono Mb ih, joesnnn Ii "MAYAK 5,55)-3- sne, t MM, E | : N a-gi g 5

C and amino-4-hydroxy»3- tov | methylcyclohexy) 74 E Сg and: 435.0 0.6 I , and | mo pyridin-3-yl)-6-(2-fluoro-5-methylphenia) picolinamidon; SHI M(4-(1К38,45,55)-3-і ож ММ, і . . ! tru she a amino-4-hydroxy-5-і denі kn methylіinclohexyl) 75 ! ZerMoy t 439.2 0.57 ,

I o pyridin-3-yl)-5-fluoro-6-(2-fluorophenyl)nicolanamidon C ee po B MA4-(1B3Ka5,55)-3-

Nad MH, Ho, y t E E Y amino-4-hydroxy-5- ! i Guy And i

She Yi Iii methylcyclohexyl) 76 was 439.2 0.55 and mo | : pyridin-3-yl)-6-(2,6- difluorophenyl) ! picolinamide !

They ate flax and N

Example o The structure of GRH/MS | РХ/MS Chemical name. / 00000000,

Me (MAN pri | (K pri

ORSI) ALL t dit M-(4-(418,35,55)-3- and She: nut as | vmino-5-' cbu and methylcyclohexyl) 77 and 441,35 0,62 , o scha B | tridin-3-l-o-(2,4. difluorophenyl)-5- and | | fluoropicolinamide and Shchy Khratenyi t NAME su E amino-4-

I I n Y gi oxypyclohexyl itzi u 425.2 0.52 what J )

C o pyridin-3-yl)-6-(2,6-difluorophenyl)licolinamide recral M-4-(18,385)-3-yl C

And aminocyclohexyl!

AK

79 her 4272 0.58 pyridin-3-yl)-6-(2,3- r. - y | T difluorophenyl)-5- fluoropicolinamide z pinin A To EK

NM t fi : 2 no E E | aminocyclohexyl) and 7.2 I pi -3-yl-6-(2,6- r. | 427.2 0.54 pyrndin-3-yl)-6-(2,6

FI be difluorophenyl)-3- ' fluoropicolinamide

EP sling M-(44K,353-3-: Na i z | y : z E aminocyclohexyl) dune 81 rUve 427.2 0.57 pyridine-3-yl)-6442.5- still - se i zn / 5 | difluorophenyl)-5-fluoropicolinamide

I am Hralny | ОМА35)3- nm З z | aminocyclohexyl) (82 bod in 427.2 0.58 pyridin-3-yl)-6-(2,4-y Azh A

FI K difluorophenyl)-5-fluoropicolinamide

Example | Structure of RC/MS SRHYUMO Chemical name. !

Me : (MAN pri | (KEpri i : i i ! : GO ORSI) MRS ny shi, z khivvlena | "MY 345)-3-

OA mn and o.

FI she | and amino-4-

In I | And hydroxycyclohexyl) 53 va a 407.1 0.51 | SHY m in pyridin-3-yl)-6-(2-! : ! and fluorophenyl) : | : picolinamide

Sleep surtayut shi O7M-4chY 345) and to. .MmN - «

C E amino-4- or hydroxycyclohexyl) and M u. ; : oxexyl) 84 Suns 465.2 0.62 | Sho m | pyridin-3-yl)-6-(2-fluoro-5-propoxyphenyl) picolinamide

Pn given OBEMYA4K3KYA555)-3- ! bark CHA H ek. | at. . ! and SE in | | amino-4-thihydroxy-5-

I Sai E | and ' nn her | methylcyclohexyl) 85 Sulem 457.2 0.56 | What can I do? pyridine-3-1l)-6-(2,6»-difluorophenyl)-5-fluoropicolinamide son sho KZ Khralnyi t M-(4-18,3B,45,55)-3- ns MH, A o - : . with TI: in: amino-4-hydroxy-5- : in : : vt : methylniclohexyl) 86 gum 457.0 0.56 mo | pyridin-3-yl)-6-(2,6-! : : i ' : : : difluorophenyl)-5- i ' fluoroticolinamide

HE YOU | M-(4-(iV3K,451-3- stitches OA TE : amino-4- and with menya ! | :

In | ND hydroxycyclohexyl) ( 87 En 425.1 0.52

Example | Structure | ех/MS РХ/MS Chemical name me (MEN at | (BE at

Re) ORSI) he in Hralny! M-(4A-(15,25,4K)D-3-

Su" E | amino-4- - eE | y

N Ch ii Y | hydroxycyclohexyl) 88 c x | 443.0 0.53 , , s Her 5 | pyridin-3-yl)-6-(2,6-difluorophenyl)-5-fluoropicolinamidon | M-(4-(3-amino-4-

Nama is, . hydroxy-5-

And methylcyclohexyl) 89 Z o b 427.3 0.63 I , ! mo pyridin-3-yl)-6-

AND

! cyclohexyl-5-fluoropicolinamide and ATX | 3-amino-M-(4-(3-amino-4-

And Well sleep, WE: hydroxy-5-no And! methylcyclohexyl) 90 : bi Moy 424.3 mb o m, pyridin-3-yl)-6-cyclohexylpicoline-

M-(4-(3-amino-5- nm sn, | :

I | methylcyclohexyl) (chi i 411.3 0.67 pyridin-3-yl)-6- zy 6 | cyclohexyl-5-fluoropicolinamido on | Z-amino-M-(4(trane)-3- and nm ! " E E | amino-4-no | hydroxycyclohex- I 92 ! and it 438.3 0.51 | and mo kv, enilinridin-3- yl)-6-I (2,6-difluorophenyl) picolinamide in C 3-amino-M-(4-(cis)-3-2; and

CODE | | amino-4- 93 no M | 438.3 0.51 y shchy M ikh | hydroxycyclohex-1-

Mo M, /enyl)pyridin-3-yl)-6-

Example Structure. TRYUMO YIRYUMS Chemical name

Heh | (MAN at | (KE at

ORSI) ORSI) ! (2,6-difluorophenyl) picolinamidon 3-amino-M-(4-(3-amino-4- nase i MH -

S E E | hydroxy-5" : H s Y | methylcyclohexyl) 94 SU t 454.1 0.54 y , - ! mo Mn, pyridin-3-yl)-6-(2,6- difluorophenyl) picolinamide

Compound 71 3-amino-M-(44(3-amino-4-.eru E hydroxy-5-ton methylcyclohexyl) 95 Sg and 454.3 0.54 and she ! mo pn, pyridin-3-yl)- 6-(2,6- difluorophenia) picolinamide мн ши Е Е amino-4-hydroxy-5-

IS

M and | and n | methylpiperidine-1- br cha 458.1 0.54 Pt s Y 5 ylupyridin-3-1ya)-6-(2,6- difluorophenyl)-5- fluoropicolinamide tone may 1 Z vmino-KiDYAZ amino-4-

NS in . and iij E hydroxy-5- i h z

A yy - methylcyclohexyl) 97 r 454.1 0.55 y ,

M Mn, pyridin-3-yl)-6-(2,6-difluorophenyl)

Sh | picolinamide

Example | Structure РХ/MS 01 вХ/MS Chemical name m (MAN at )(KEpr

PORSI) ORSI) he Chiral 3-amino-M-(4-(3-amino-4-peru E hydroxy-5-

There are tons of methylcyclohexyal)

M M 454.1 0.54 , I and pyridine-3-1l)-6-(2,6-54 o Mn, m difluorophenyl) picolinamide

Khirelny -(4-(1K,35)-3-

PUFI and M-(4-((1K,35)-3

I i i: aminocyclohexyl) 99 g M pe 427.2 0.55 pyridin-3-yl)-6-(2,6-difluorophenyl)-5-fluoropicolinamide

I Chiral 3-amino-M-(4-(1K,353-3-su" SI aminocyclohexyl)

And E

100 that 442.2 0.59 pyridin-3-yl)-6-(2,6- i | | | and

FI Mn, | difluorophenyl)-5-fluoropicolinamide o | 2-amino-4-4(3-(3-amino-6-moene, De | 2,6-difluorophenyl)

MN J | (2,6-difluorophenyl 101 C sho 482.2 0.56 Pi no | picolinamido)pyridine-4-

S o kv, | yl)ucyclohexylanetatone Z t Z-amino-M(4-3-amino-y-y ach i

SEE E hydroxycyclo- 102 places | 440.3 0.52 hexylinridin-3-yl)-6-

FI mn, (2,6-difluorophenyl) picolinamide on kharapny | Z-amino-M-(4- er in | ((3K.48,55)-3-amino-4-

Se Ii r | hydroxy-5- 103 | we about mn, 455.3 0.53 methylpiperidin-1-yl)pyridin-3-yl)-6-(2,6-difluorophenyl)

Picolinamide

Example / Structure of RYU/MS RH/MS Chemical name in (MAN with |((КЯ with

IRSI) PRSI) he in | 3-Amino-M-(4-pyridin-3-yl)-6-62,6- .. difluorophenyl) picolinamide ! 3-amino-6-(2,6- difluorophenyl)-M-(4-(3-re / hydroxy-5- 105 ich: ny y y 438.2 2.92 fo m, | methylcyclohex-1- enyl)pyridin-3-yl)picolinamide

3-amino-M-(4-((1K,35)-3-min, aminocyclo- 106 Xa Ii p 394.3 0.74 hexyl)pyridin-3-yl)-6-z o Mn, niclohexylpicolin-amide we, E aminocyclohex-1- 107 I 421.9 0.59 enyl)pyridin-3-yl)-6-ia b m, (2,6-difluorophenia) ! Picoliniaamide that NEZI M-A(4-(1B,3,55)-3-

Y and amino-5-105 and B. A, | (trifluoromethyl)cyclohex

C 47343 0.67 m syl)niridin-3-yl)-6-(2-fluoro-5-methylphenyl)

Picolivnamid o azhevalny | M-(44(1К3К,55)-3- in. ХО Her |amino-5- 109 and F and E 507.2. 0.65 I (trifluoromethyl)cyclo-s I chu | hexyl)pyridin-3-1l)- 6-

FI about | (2,6-difluoro-4- and methoxyphenyl) !

Example | Structure of РХ/MS РХ/MS | Chemical name

Me (MAN at |((K at all) MRSI)

And that's it

Zdu, Zhralnyi Z-amino-M-(4-.

EX MN, | I and I and eat

P ti E E (1K.UK,55)-3-amino-5- po niio I 510.2 0.64 (trifluoromethylcyclo-

In s ze with . . . and: ; and hexyl)pyridine-Zl1l)-b-

Oh my,

In, - g

M (2,6-difluorophenyl)-5-fluoropicolinamide and Chiral M-(4-(18,38,55)-3-

K.I Mn, 7th amino-5-

F!

PI N M (trifluoromethyl)cyclo-ho. ik 465.3 0.72

ST | | hexyl)pyridine-3-1l)-6- and | diclohexyl-5-fluoropicolinamide

Shit; g Horsetail Z-amino-M-(4-

What else are we doing? (1K.3K,55)-3-amino-5- 12 tn | (trifo methylucyclo-

C t 4922 0.2 | "P ? and cho p, hexyl)pyridin-3-yl)-6-(2,6-difluorophenyl)

Picolinamide on he Heralny | M-(4-(435,45,583-3-er) and amino-4-hydroxy-5-113 and I and ! more | methylpiperidine-1-

Su 458.2 0.55 ШЙ pn sho yl)pyridin-3-yl)-6-(2,6- difluorophenyl)-5- fluoropicolinamide on yellow | M-(4-(38.48,583-3,4- 4 che on E | dihydroxy-5-

M" c h- E 44 0.61 methylpiperidin-1-yl)pyridin-3-yl)-5-fluoro- z o 6-(2-fluorophenyl)

Example | Structure of Рх/MS РХ/MS Chemical name

Me (MAN at | (EC at !

ORSI) OVS) on Chiral | 6-(2,6-difluorophenyl)-M-nsia sut E (4-(3K,4K,55)-3,4- 115 dihydroxy-5-

M nm 441 Ro g same methylpiperidine-1-zm | oyl)pyridin-3-yl)picolinamidon Fatty 6-42,6-difluorophenyl)-M- ut E (4-((38,48,55)-3,4-

IS

M and h | dihydroxy-5- g and 459.1 0.61 and sia o methylpiperidin-1-yl)pyridin-3-yl)-5- fluoropicolinamido on ton o 0/ Chosen | | 6-(2,6-Difluorophenyl)-M-esutya ak (444183 B,48,55)-3,4- 117 c EM Y and dihydroxy-5-chu 458.2 0.62

Su methylcyclohexyl) nsya o

M pyridin-3-yl)-5-fluoropicolinamide with Fnortnia M-(4-(3B4K,55)-3-neruya de amino-4-hydroxy-5-

IS

118 M, methylpiperidine-1-in soap 423.2 0.32 MeTHITIRach-1 8 yl)pyridin-3-yl)-3-fluoro-m: 2,3-bipyridine n-6-carboxamide

I. OO M(4-(38,4B,55)3-3-. flows "M in E . -

E amino-4-hydroxy-5-119 M M Y su" B 441.2 0.46 methylpiperidin-1- mol)pyridin-3-yl)-3,3'" difluoro-2,4-bipyridine-b -

Example | Structure /РХx/МСо РХ/Mo Chemical name

Me (MN at | (VE at

ORSI) ORSI) at 2 | M-(4-4(384K,55)-3.thum and amino-4-hydroxy-5- 120 M M and | methylpiperidine-1- and Muaz 0.3 shi o

SHIE 2 | yl)upyridin-3-yl)-3-fluoro- and 2,4-bipyrides n-6-carboxamide and Fatty 3-amino-6-(2 "b- nas, x OH E difluorophenyl)-M-(4. - company Y TSIV, ZK, 48, 55)-3,4-

I21 from Mich

C 473, 0.68 dihydroxy-5-

M o nn, methylcyclohexyl) pyridin-3-yl)-5- fluoropicolinamide

Synthesis of 6-bromo-N-(4-(3-tert-butyldimethylsilyloxy)-5-methylcyclohex-1-enyl)pyridin-3-yl)-5-fluoropicolinamide tvoM5o

Hg

IS

M

Ch d M xx "L o

M

According to method 9, 4-(3-(tert-butyldimethylsilyloxy)-5-methylcyclohex-1-enyl)pyridin-33-amine and 6-bromo-5fluoropicolinic acid were introduced into the reaction of the combination and after adding EOAc and washing with H2H2O, MaSiVza) After drying over Mo9505, b6-bromo-N-(4-(3-(tert-butyldimethylsilyloxy)-5-methylcyclohex-1-enyl)pyridin-3-yl)-o-fluoropicolinamide was obtained. RHMS (t/;): 455.3 (MNU; RH Ke2.09 min.

Synthesis of 6-bromo-M-(4-(1B, 35)-3-(1,3-dioxoisoindolin-2-yl)y-cyclohexyl)pyridin-3-yl)-5-fluoropicolinamide th

M

Vg is); And

BUT

In c)

M

According to method 9, 2-(3-(33-aminopyridin-4-yl)ucyclohexyl)isoindoline-1,3-dione and b-bromo-5fluoropicolinic acid were introduced into the reaction of the combination and after addition of EtOAc and washing with Neo,

Mass and drying over Mo95O45 gave 6-bromo-N-(4-(18, 35)-3-(1,3-dioxoisoindolin-2-yl)ucyclohexyl)pyridin-3-yl)-5-fluoropicolinamide. RHMS (t/lg2): 523.2/525.2 (MNU); РХ 3.31 min.

Synthesis of 3-amino-6-bromo-M-(4-((18, 35)-3-(1,3-dioxoisoindolin-2-yl cyclohexyl)pyridin-3-yl)-5-fluoropicolinamide i and l

Vg is); And

BUT

In about Mn

M 2

According to method 9, 2-(3-(3-aminopyridin-4-yl)ucyclohexyl)isoindoline-1,3-dione and 3-amino-6-bromo-5-fluoropicolinic acid were introduced into the combination reaction and after adding EAs and washing with 3-amino-6-bromo-M-(4-(18, 35)-3-(1,3-dioxoisoindolin-2-yl)ucyclohexyl)pyridin-3- yl)-5-fluoropicolinamide. RHMS (t/g2): 538.1/540.1 (MNU; RH

In 3.46 min.

Synthesis of tert-butyl-(15, ZA, 55)-3-(3-(6-bromo-5-fluoropicolinamido)-pyridin-4-yl)-5-methylcyclohexylcarbamate

Resurrection

Hg

E and no where M Zh -75

M

According to method 9, tert-butyl-(15, ZA, 55)-3-(3-aminopyridin-4-yl)-o-methylcyclohexylcarbamate and 6-bromo-5-fluoropicolinic acid were introduced into the combination reaction and after adding EOAc and washing with using HgO, MasSivad and drying over M95Ox gave tert-butyl-(15, ZA, 55)-3-(3-(6-bromo-5-fluoropicolinamido)pyridin-4-yl)y-5-methylcyclohexylcarbamate. RHMS (t/2): 5071/5091 (MNU), B-0.90

MIN.

Synthesis of (Kk, 2K, Ak, 65)-4-(3-(6-bromo-5-fluoropicolinamido)pyridin-4-yl)-2-(tert-butoxycarbonylamino)-b-methylcyclohexylacetate

OAS

Vosnm Ve d E no to M Zh zu / o

According to method 9 (1, 2K, 4, 65)-4-(3-aminopyridin-4-yl)-2-(tert-butoxycarbonylamino)-6-methylcyclohexyl acetate and b-bromo-5-fluoropicolinic acid were introduced into the reaction of the combination and after adding EOAc and washing with HgO, Mas(za) and drying over MaSO»x, (18, 2B,

AB, 65)-4-(3-(6-bromo-5-fluoropicolinamido)pyridin-4-yl)-2-(tert-butoxycarbonylamino)-6-methylcyclohexylacetate. RHMS (t/2): 567.2 (MNU), NeO, 82 min.

Synthesis of (1Y-)-tert-butyl-5-(3-(6-bromo-5-fluoropicolinamido)pyridin-4-yl)-7-methyl-2-oxohexahydrobenzo|F|oxazole-3(2H)-carboxylate

F)

Ho

VosM

Vg d E not to | M sx to c)

M

According to method 9 (h/-)-tert-butyl-5-(3-aminopyridin-4-yl)-7-methyl-2-oxohexahydrobenzo|4|oxazol-

C(2H)-carboxylate and 6-bromo-5-fluoropicolinic acid were introduced into the combination reaction and after adding

EtOAc and washing with HgO, MasCyzase and drying over M95O»x gave (-/-)-tert-butyl-5-(3-(6-bromo-5-fluoropicolinamido)pyridin-4-yl)-7-methyl- 2-oxohexahydrobenzo|chCh|oxazole-Z(2H)-carboxylate. RHMS (t/lg): 5492/5512 (MNU), Ne0.78 min.

Synthesis of tert-butyl-5-(3-(6-bromo-5-fluoropicolinamido)pyridin-4-yl)-2-oxohexahydrobenzo|chCh|oxazole-

C(2H)-carboxylate c) 0-X

M-Vos

Vg ai E no -/75

M

According to method 9, tert-butyl-5-(3-aminopyridin-4-yl)-2-oxohexahydrobenzo|H|oxazole-3(2H)-carboxylate and 6-bromo-5fluoropicolinic acid were introduced into the reaction of the combination and after adding EtOAc and washing tert-butyl-5-(3-(6-bromo-5-fluoropicolinamido)pyridin-4-yl)-2-oxohexahydrobenzo|H|oxazole-Z(2H)- carboxylate. LCMS (t/lg): 537.1 (MNU; LCMS K-0.71 min.

Synthesis of 6-bromo-M-(4-(18,5H8)-5-(1,3-dioxoisoindolin-2-yl)-3,3-dimethylcyclohexyl)pyridin-3-yl)-5-fluoropicolinamide i i i

Vg o E and no o

According to method 9, 2-(1H, 5H8)-5-(3-aminopyridin-4-yl)-3,3-dimethylcyclohexyl)isoindoline-1,3-dione and 6-bromo-5-fluoropicolinic acid were introduced into the reaction of the combination and after addition of EOAc and washing with H2O, NaCl and drying over MozOi, b-bromo-M-(4-(18, 5H8)-5-(1,3-dioxoisoindolin-2-yl)-3,3-dimethylcyclohexyl was obtained )pyridin-3-yl)-5-fluoropicolinamide LCMS (t/g2): 551/553 (MNU), Vie0.95 min.

Synthesis of M-(4-(1НВ, ЗВ, 45, 58)-3,4-bis(tert-butyldimethylsilyloxy)-5-methylcyclohexyl)pyridin-3-yl)-6-bromo-5-fluoropicolinamide

Otv5 tv5o am

Hg

Oh and but)

AND

M

According to method 9, 4-(18, ZA, 45, 58)-3,4-bis(tert-butyldimethylsilyloxy)-5-methylcyclohexyl)pyridin-3-amine and b6-bromo-5fluoropicolinic acid were introduced into the combination reaction and after adding EOAc and washing with HgO, MaSi(zao and drying over MoaZOx gave M-(4-18, ZB, 45, 58)-3,4-bis(tert-butyldimethylsilyloxy)-5o-methylcyclohexyl)pyridin-3-yl)- 6-bromo-5-fluoropicolinamide. RHMS (t/y): 652.5, 652.4 (MNU); РХ K-5.82 Min.

Synthesis of M-(4-(15, 35, 48, 55)-3,4-bis(tert-butyldimethylsilyloxy)-5-methylcyclohexyl)pyridin-3-yl)-6-bromo-5-fluoropicolinamide

ОтТв5 т, ОтТв5 Ве

And no

ZM Zh and 5

M

According to method 9, 4-(15, 35, AB, 55)-3,4-bis(tert-butyldimethylsilyloxy)-5-methylcyclohexyl)pyridin-3-amine and 6-bromo-5fluoropicolinic acid were introduced into the combination reaction and after adding EOAc and washing with HgO, MaSi(zao and drying over MoaZOx gave M-(4-(15, 35, 48, 55)-3,4-bis(tert-butyldimethylsilyloxy)-5o-methylcyclohexyl)pyridin-3-yl) -6-bromo-5-fluoropicolinamide. RHMS (t/y): 652.5, 652.4 (MNU); РХ Ke5.83 min.

Methodology 10

Synthesis of 6-(2,6-difluorophenyl)-5-fluoro-N-(4-(3-hydroxy-5-methylcyclohex-1-enyl)pyridin-3-yl)upicolinamido

E E

Oh well

In c)

M

A solution of b-bromo-M-(4-(3-(tert-butyldimethylsilyloxy)-5-methylcyclohex-1-enyl)pyridin-3-yl)-5-fluoropicolinamide (1.0 equiv.), 2,6-difluorophenylboron acid (3.0 equiv.), tetrakistriphenylphosphine (0.2 equiv.) and triethylamine (3.0 equiv.) in a 1:11 EFN/toluene (0.1 M) mixture was heated at 120 C by microwave radiation for 1200 s. Upon cooling, the volatiles were removed in vacuo, and the product of the Suzuki reaction was immediately purified by reverse-phase HPLC. The fraction containing the product was lyophilized and the protective group was removed from the obtained TVOM5 ether as described in method 9, and after purification by ZF HPLC and lyophilization, 6-(2,6-difluorophenyl)-5-fluoro-M-(4 -(33-hydroxy-5-methylcyclohex-1-enyl)pyridin-3-yl)picolinamide in the form of a salt with TFC. RHMS (t/2): 438.2 (MH; RH Ki-2.00 min.

Synthesis of M-(4-((1B, 35)-3-aminocyclohexyl)pyridin-3-yl)-6-(2,6-difluorophenyl)-5-fluoropicolinamide

NhM 7 E E

E and no d M ХУ -/Л 5

M

A solution of b-bromo-M-(4-((18, 35)-3-(1,3-dioxoisoindolin-2-yl cyclohexyl)pyridin-3-yl)-5-fluoropicolinamide (1.0 equiv.), 2, 6-difluorophenylboronic acid (3.0 equiv.), tetrakistriphenylphosphine (0.2 equiv.) and triethylamine (3.0 equiv.) in a mixture of 1:11 ETH/toluene (0.1 M) was heated at 120 C by microwave radiation for 1200 s. On cooling, the volatiles were removed in vacuo, the product of the Suzuki reaction was immediately purified by reverse-phase HPLC. The fraction containing the product was lyophilized, and the resulting phthalimide was deprotected as described in Method 9 and after purification by ZF HPLC and lyophilization gave M-(4-(18, 35)-3-aminocyclohexyl)pyridin-3-yl)-6-(2,6-difluorophenyl)-5-fluoropicolinamide in the form of a TFC salt. RHMS (t/2): 427.2 (MN); РХ K-2.26 min.

Synthesis of 3-amino-M-(4-(18, 35)-3-aminocyclohexyl)pyridin-3-yl)-6-(2,6-difluorophenyl)-5-fluoropicolinamide

NhM 7 E E

IS

M

What about Mn

M 2

A solution of 3-amino-6-bromo-M-(4-((18, 35)-3-(1,3-dioxoisoindolin-2-yl cyclohexyl)pyridin-3-yl)-5-fluoropicolinamide (1.0 equiv. ), 2,6-difluorophenylboronic acid (3.0 equiv.), tetrakistriphenylphosphine (0.2 equiv.) and triethylamine (3.0 equiv.) in a mixture of 1:11 ETH/toluene (0.1 M) was heated at 120 with microwave radiation for 1200 s. On cooling, the volatiles were removed in vacuo, the product of the Suzuki reaction was immediately purified by reverse-phase HPLC. The fraction containing the product was lyophilized and the resulting phthalimide was deprotected as described in Method 9 and after purification by 3-amino-M-(4-(1B, 35)-3-aminocyclohexyl)pyridin-3-yl)-6-(2,6-difluorophenyl)-5-fluoropicolinamide was obtained in the form of a salt from TFC using ZFf HPLC and lyophilization . RHMS (t/2): 442.2 (MN); РХ -2.24 min.

Synthesis of M-(4-(33-amino-4-hydroxycyclohexyl)pyridin-3-yl)-6-(2,6-difluorophenyl)-5-fluoropicolinamide

N

NhM 7 E E

IS

M i xx c)

M

Solution of tert-butyl-5-(3-(6-bromo-5-fluoropicolinamido)pyridin-4-yl)-2-oxohexahydrobenzo|H|oxazole-

C(2H)-carboxylate (1.0 equiv.), 2,6-difluorophenylboronic acid (3.0 equiv.), tetrakistriphenylphosphine (0.2 equiv.) and triethylamine (3.0 equiv.) in a 1:11 mixture ETH/toluene (0.1 M) was heated at 120 C by microwave radiation for 1200 s. Upon cooling, the volatiles were removed in vacuo, and the product of the Suzuki reaction was immediately purified by reverse-phase HPLC. The fraction containing the product was lyophilized and the Bos protecting groups were removed from the obtained cyclic carbamate as described in method 9, and after purification by ZF HPLC and lyophilization M-(4-(3-amino-4-hydroxycyclohexyl)pyridine-3 was obtained -yl)-6-(2,6-difluorophenyl)-5-fluoropicolinamide in the form of a salt with

TFC RHMS (ti/): 443.2 (MNY); RH G-2.11 min.

Synthesis of B-amino-M-(4-(18, 35)-3-aminocyclohexyl)pyridin-3-yl)-3,3'-difluoro-2,4-bipyridine-6b-carboxamide

M

WITH

NhM -d E

E ai no heh oh MN

M 2

A solution of 3-amino-6-bromo-M-(4-((18, 35)-3-(1,3-dioxoisoindolin-2-yl cyclohexyl)pyridin-3-yl)-5-fluoropicolinamide (1.0 equiv. ), 3-fluoropyridin-4-ylboronic acid (3.0 equiv.), tetrakistriphenylphosphine (0.2 equiv.) and triethylamine (3.0 equiv.) in a mixture of 1:11 EHN/toluene (0.1 M) was heated at 120 "C with microwave radiation for 1200 s. When cooling, volatile substances were removed in vacuo, the product of the Suzuki reaction was immediately purified using reversed-phase HPLC. The fraction containing the product was lyophilized and the protective group of the resulting phthalimide was removed as described in method 9, and after purification by ZF HPLC and lyophilization, 5-amino-M-(4-(18, 35)-

3-Aminocyclohexyl)pyridin-3-yl)-3,3'-difluoro-2,4-bipyridine-6-carboxamide as a salt with TFC. RHMS (t/2): 425.1 (MN); РХ K-2.08 min.

Synthesis of M-(4-Ch18, Z5, 55)-3-amino-5-methylcyclohexyl)pyridin-3-yl)-6-(2,6-difluoro-3-hydroxyphenyl)-5-fluoropicolinamidon

Nam E E

E no s M y r

M c)

To a solution of tert-butyl-(15, ZA, 55)-3-(3-(6-bromo-5-fluoropicolinamido)pyridin-4-yl)-5-methylcyclohexylcarbamate (1.0 eq.) in a microwave-safe dish 2,6-difluoro-3-hydroxyphenylboronic acid (5.0 equiv.), KE (5.5 equiv.) and Rag(aba)z (0.2 equiv.), then THFf and water (10:1, 0.03

M). To this mixture was added P(I-Vy)z (0.4 equiv.) and the reaction mixture was heated in a microwave oven at 100 "С for 30 min. Then the organic phase was separated, the aqueous layer was washed with ethyl acetate, and the organic substances were combined and concentrated in vacuo . -5-methylcyclohexyl)pyridin-3-yl)-6-(2,6-difluoro-3-hydroxyphenyl)-5-fluoropicolinamide In the form of a salt with TFA. LCMS (m/2): 457.2 (MH); LC 2.17 min.

The following compounds were obtained according to method 10:

Table 2

Example M Structure of TRHMO TRHMS | The chemical name of the identity" (MN (Kgpr

And fications at all

Me MUR ORSI)

ON. chiral M-4-(1K058U5-amino- peresa : : 3.3-pimethylcyclo- and sy!i : g shield H : sc tutuzchi Kg

And! there is h Ei | ! texyl)pyridin-3-yl)-6- 122 storm but would throw | (2,b-difluoro-4-; E: tihydroxyphenyl)-5" fluoropicolinamide

KUYE in Zhralnia M-(4-41K,58)-5-aminori | and 3.3-dimethylcyclones cl o | | sh n ve 5 SHY B. hexylpridin-Z-p)-b-

I23 | tone 4852065

Yaru (2,6-difluoro-4- zm I methoxyphenyl)-5-

With GU and fluoropicolinamide, ZNChral MA4-(1K,35)-3-:

Y I nk /f | | ( ( » 3

Nv che wl ze ; Н захгетутяя к : К | Her ! changepiclohexyl) ! Name: lower pyridin-3-yl)-6-(2,6-24 | Su B 443.2 0.54 umo difluoro-3-hydoxyphenium)-5-fluoropicolinamidX rbannkn MIK 5U-

Ah - : T amino-5-methylnickel-

N y gi 3-ip)-6 ' | A M i hexyliniridin-3-yl)-6- 125 shi fe 457.2 0.57 ! with (2.b-lifluoro-3-hydroxyphenia)-3-

No

"Nryklad Me/ | Structure РХ/MS РХ/MS Chemical name! identities- | (МНО 0 (Вгпрн) fication | at PRSI) | :

Ho MUR | MRS pork nin nn you What but ; DM of the day of knowledge of Zhe MAYA VK K55)53 : ) E I eV AN A N . I am N

Another g Z KE amino-4-hydroxy-5-re a Mmethylinclohexnia) I

C and zhi 7! 126 a 507, 0.65 pyridin-3-yl)-5-fluoro-b»!

O-fluoro-5- : : ((xtrifluoromethylphenia) : picolinamim

Y N N

TE A Z pit TT vn pe SE toregens and M-(4-tsi K.ZEYAK,55)-3- ne, , "MN, | and k x ! and shk: amino-i-hydroxy-5- and n "sha din! and SI or methylcyclohexnia) i27 mo 507.1 10.65 pyridine-Z-id 1-5 fur

O-fluoro-3- ! : | "(trifluoromethylphenyl) ! picolinamide ! and vin nn nn schi ptn re МА41В,35,55)-3- : What s, ery, amino-5- ! ! and what what ! and X what M | methnlcyclohexnl) and I 4 and p zho | and tni 125 zr o 457.2 10.58 shiridin-Z-yl)-b-(2,6e! difluoro-4-hydroxyphenyl)-3- tovpicolinamide!|! p! KS! M4-i K. 35.55)-3-ny SEN oh i; tt v Vo i | amino-5-

N ks and Mch same

Am | mMmethylcyclohexyl) 1729 AH Lo 459.2 0.62 | and zm o and shiridin-2-yru- 5-Fftor-b- ! ! 2,3 bltrifluorophenyl) (picolinamide

Example L/ | Structure of Vzh/Me 1РХ/MS | The chemical name of the identi- Man IC at fication lira.RSE

Me MUR SRS t pe u shi shsh tu samino- Ma saminoi ov put y | 4-hydroxy-5- 130 pluku 4731 0.56 Matt t

M z tridin-3-111-60-45,0- : difluorophenyl)-5- fluoropicolinamide 77 3-amino-M-(4-(3-amino-

It is a 4-tihydroxycyclo- and 131. KAH 458.0 (0.57 texyl)pyriline3511)-0- and a M, | (2b-difluorophenyl)-5-fluoropicolinamide. then What is in" 4-hydroxy"5- about | I and A | | | methylnicklohexyl)

I32 neck 473 Sh te

Ge 2 | pyridine-z-1l)-0-5,0" difluorophenyl)-5-fluoronicolinamide

And Shv | 3-amino-M-(4-(3-amino» ru t Y | | /5-methylcycle- 133 | xx A (456.3 (0.60 ron still o nn | | (2,b-difluorophenia)- 5-

Her | o" fluoropicolinamide: - roz ot sh sasa 134 ! shsh p acty 057 methylcyclohexy) y | pyridin-3-yl)-6-2,6- difluorophenyl)-5- sh | fluoropicolinamide More

She; ryklad Me/ | It is structured РХ/Me AND РХМе | Chemical name. ШЩ педенти- NOUNS WHAT kind of pre

Vikapiinny | don't worry

PM MUuv | PORSI) p in pennnrnnnntnya knife j nich u an : he I | M-4-13-amino-4-s in | tidroxy-5-nn | | and ryh efki tiru bug zhi si i ! methylinclohexyl) 135 of them "7 457.1 10.55 NI and ! with: pyridine-3-1l)-6-(2.65% difluorophenyl)-α-fluoromnicolinamyl

HE, well, sh re y! | Z-vminocyclone NI shaeny. IS

HELL. DT | hexylipyridin-Z-ylin oC

IZ6 | S GMO M dz 046 vk OM, ! 3, with "difluoro-2 - ! ; ! : | biviridine-b" carboxamide sira nni nn a a nn a and schi 6-2, b-difluorofetyl-ia"

Pd ONreboya kr! how do you know: 00 deermochidyukon! di I 440.2 0.75 and 5-methylcyclohex-1-! with | | enyl)ipyridine-3- : s ilupicolinamid ' on Vi Mrelnya | 6-2,b-difluorophenyl)Me

But SN yat! and; that preshi and (4418538.45,58)-3,4- are not

IZ8 r, IY dihydroken-ya» obu» 458.2 |0.64 with Z : :methylniclohexyal) ! pyrilin-I-l r I- and no! and ! fluoropicolinamide

Example 139

Synthesis of M-(4-(18, ZA, 45, 55)-3-amino-4-fluoro-5-methylcyclohexyl)-pyridin-3-yl)-6-(2,6-difluorophenyl)-5-fluoropicolinamide

IS

NhM 7 E E aka

Ch hu M - 6)

M

To a solution of tert-butyl-(18, 28, 35, 58)-5-(3-(6-(2,6-difluorophenyl)-5-fluoropicolinamido)pyridine-4-

yl)-2-hydroxy-3-methylcyclohexylcarbamate (1.0 equiv.) in DCM (0.04 M) at 0 °C was added DATS (1.0 equiv.). The reaction mixture was stirred for 1.5 h at 0 "C, then added to the reaction mixture

TFC (10 equiv.). After 2 hours the reaction mixture was concentrated in vacuo and the residue was purified by preparative HPLC to obtain M-(4-(1B, ЗВ, 45, 55)-3-amino-4-fluoro-5-methylcyclohexyl)pyridine-

3-yl)-6-(2,6-difluorophenyl)-5-fluoropicolinamide as a salt with TFC. RHMS (t/h2): 459.3 (Mn); РХ -2.39 min.

In addition to LC/MS and LC characterization, typical compounds were analyzed by H-NMR. Below are typical spectra of compounds proposed in this invention.

Example 4

Salt from NSI, 1H-NMR (400 MHz, dmMso-ab): a 10.54 (p, 1H), 8.80 (B5, 1H), 8.55 (y, 1H), 8.34 (aa, 1H),

BO 8.20 (I, 1H), 8.00 (65, 2H), 7.69 (t, 1H), 7.56 (a, 1), 7.34 (i, 2H), 3.10- 3.0 (t, 2H), 2.83 (t, 1H), 2.03 (y, 1H), 1.76-1.59 (t, 2H), 1.40 (t, 1H), 1 .31-1.21 (t, 1H), 0.92 (y, ЗН).

Salt from NSI, IN-NMR (DMSOSO-a6): Aa 10.4 (5, 1H), 8.71 (5, 1H), 8.37 (a, 1H), 8.14-8.20 (t , 2H), 7.94 (p5, 2H), 7.86-7.88 (t, 2H), 7.54-7.58 52 (t, 1H), 7.30 (a, 1H), 7 ,22-7.26 (t, 2H), 2.09-3.02 (t, 2H), 2.78 (t, 1H), 1.96-1.99 (t, 1H), 1.68 -1.71 (t, 1H), 1.60 (d, 1H), 1.37 (t, 1H), 1.15-1.24 (t, 1H), 0.88 (y, ЗН

Salt from NSI, 1H NMR (400 MHz, dmMeoo-ab): a 10.59 (5, 1H), 8.92 (5, 1H), 8.62 (a, 1H), 8.37 (da, 1H ), 8.23 (, 1H), 8.19 (p, 2H), 7.68-7.71 (t, 70 2H), 7.36-7.40 (t, 2H), 3.01- 3.10 (t, 2H), 2.01-2.05 (t, 1H), 1.94-1.97 (t, 1H), 1.72-1.76 (t, 1H), 1, 46- 1.53 (t, 2H), 1.01-1.13 (t, 2H), 0.89 a, ZN

Salt from NSI, IN NMR (DMSOSO-a6): Aa 10.37 (5, 1Н), 8.61 (5, 1Н), 8.41 (а, 1Н), 8.29 (da, 1Н), 8 ,13 (, 1H), 7.8 85 (b5, 2H), 7.69-7.61 (t, 1H), 7.34-7.28 (t, ЗН), 3.061 (t, 1H), 2.86 (t, 1H), 1.76-1.63 (t, 2H), 1.53-1.47 (t, 1H), 1.4-1.34 (t, 2H), 0, 82 (th,

ZN).

Salt from NSI, IN NMR (DMSOSO-a6): Aa 10.42 (5, 1Н), 8.62 (5, 1Н), 8.46 (t, 88 1Н), 8.32 (t, 1Н), 8.18 (1, 1H), 7.76 (t, 2H), 7.67 (t, 1H), 7.35 (t, ЗН), 5.33 (biv, 1H), 3.108 (t, 2H ), 2.88 t, 2H), 1.65 (t, 2H), 1.48 (t, ZN).

Salt from NSI, 1H NMR (400 MHz,

СОЗ3О0): a 9.09 (5, 1 H), 8.46 (ad, 1

H), 8.39 (aa, 1 H), 8.05 (her, 1 H), 7.57-7.67 (t, 1 H), 7.53 (9, 1 H), 7.16 - 7.25 (t, 2 H), 4.03-4.12 (t, 1 H), 3.85-3.94 (t, 1 H), 3.20 (5, C H), 2 .70-2.80 (t, 1 H), 1.67-1.79 (t, 1

H), 0.83 (9, with H). be TEUKIME MN 9.93(5, 1H), 9.38(5, 1H), 8.40-

8.45(t, 1H), 8.40(a9, 1H), 7.74-7.80(t, 1H), 7.47-7.55(t, 1H), 7.19(a , 1n), 7.06-7.13(t, 2H), 2.68-2.83(t, 2H), 1.97-2.05(t, 1H), 1.65-1.95 (t, 5H), 1.22-1.40(t, ЗН), 1.04-1.15(t, 1H).

Salt from NSI, IN NMR (DMSOSO-a6): Aa 10.13(5, 1Н), 8.825, 1Н), 8.41 (а, 100 1Н), 7.94(р5, 2Н), 7.52- 7.62(t, 1H), 7.36(a, 1H), 7.36 (p, 2H), 7.20-7.31(t, ЗН), 2.78-2.88(t, 2H), 1.70-2.02(t, 4H), 1.16-1.54(t, 4H).

Salt from NOSI, IN NMR (400 MHz, dmMeoo-ab): a 10.59 (5, 1H), 9.30 (5, 1H), 8.54 (a, 1H) 8.08 (horn 5, ЗН ), 102 7.755a, 1), 7.65 (a, 1H), 7.60-7.56 (t, 1H), 7.49 (a, 1H), 7.33 (i, 2H), 4 .04 (horn 5, 1H), 3.16 (rg 5, 2H) 3.05 ry, 1H), 1.98-1.20 (t, 7H

Salt from NSI, 1H-NMR (400, ab-

DMS): a 10.47 (5, 1H), 8.56 (5, 1H), 8.33 (aa, 1H), 8.26 (aa, 1), 116 8.20 (B 1H), 7 ,62-7.72 (t, 1H), 7.30-7.35 (t, ЗН), 3.82-3.92 (t, 2H), 3.18-3.22 (t, 1H) , 2.84-2.91 (t, 1H), 2.69 (i, 9-13.2, 1H), 1.38-1.46 t, 1H), 0.69 (a, ЗН).

Salt from NSI, 1H-NMR (400, ab-

DMS): a 11.00 (5, 1H), 10.46 (5, 1H), 8.55 (a, 1H), 8.29 (aa, 1H), 8.15 128 (, 1H), 8 .05 (65, 2H), 7.54 (a, 1H), 6.72 (9, 2H), 3.04-3.10 (t, 1H), 2.92-3.04 (t, 1H ), 2.01 (9, 1H), 1.95 (9, 1H), 1.74 (Od, 1H), 1.42-1.52 (t, 2), 0.97-1.08 ( t, 2 H), 0.88 (а, ЗН).

Example 140

Study of ATP consumption under the influence of Rit!

RIMI1 activity is determined using a luciferase-luciferin-based reagent to quantify ATP (adenosine triphosphate) consumption due to kinase-catalyzed phosphorite transfer to a peptide substrate. The studied compounds are dissolved in 100 95 DMSO and directly placed in 384-well tablets at 0.5 μl/well. To initiate the reaction, add 10 μl of 5 nM Rit kinase to each well! and 80 µM of the peptide VAYU (KESENZZURAST-ON) in the assay buffer (50 mM NEREZ (M-2-hydroxyethylpiperazine-M-2-ethanesulfonic acid) pH 7.5, 5 mM Mass), 1 mM DTT (dithiothreitol), 0 .05 95 BSA (bovine serum albumin)). After 15 min, add 10 μl μM ATP in assay buffer. The final concentrations during the analysis were equal to 2.5 nM RIM'IA, 20 μm

ATP, 40 µM HAO peptide and 2.5 95 DMSO. The reaction is carried out until approximately 50 95 is consumed

ATP, then it is stopped by adding 20 μl of Kipazedio Riyi5 solution (Rgoteda Sogrogayop).

When the reaction is stopped, the mixture is incubated for 10 min and the remaining ATP is determined by luminescence using the Misiog2 device (RegKip EIteg). The compounds of the above examples are studied by studying ATP consumption under the influence of Rit! and find that they are characterized by IC50 values shown below in Table 3. IC50, half maximal inhibitory concentration, means the concentration of the test compound that is required to inhibit 50 95 of its target i migo.

Example 141

Study of ATP consumption under the influence of Rit2

The activity of RIM2 is determined using a luciferase-luciferin-based reagent to quantify ATP consumption due to the kinase-catalyzed phosphorite transfer to the peptide substrate. The studied compound is dissolved in 100 95 DMSO and directly placed in white 384-well tablets at 0.5 μl/well. To initiate the reaction, 10 nM Rit2 kinase and 20 µM HAO peptide (KZKNOZURAST-OH) in assay buffer (50 mM

NEREZ pH 7.5, 5 mm Massi", 1 mm DTT, 0.05 95 BSA). After 15 min, add 10 μl of 8 μM ATP in assay buffer. The final concentrations during the analysis were equal to 5 NM PIM2, 4 μM ATP, 10 μM peptide ВАХО and 2.595 DMSO. The reaction is carried out until approximately 50 95 ATP is consumed, then it is stopped by adding 20 μl of a solution of Kipazedio Rii5 (Rgoteda Sogrogayop). When the reaction is stopped, the mixture is incubated for 10 min and the remaining ATP is determined by luminescence using the device

Misyug2 (RegKip EITeg). The compounds of the above examples are studied by means of an ATP consumption study under the influence of Rit and found to be characterized by the ISvo values shown in Table 3 below.

Example 142

Study of ATP consumption under the influence of RitZ3

The activity of RIMZ is determined using a luciferase-luciferin-based reagent for the quantification of ATP consumption due to the transfer of phosphorite catalyzed by the kinase to the peptide substrate. The studied compound is dissolved in 100 95 DMSO and directly placed in white 384-well tablets at 0.5 μl/well. To initiate the reaction, 10 μl of 10 nM Rit3 kinase and 200 μM of the HAO peptide (K5KNZURAST-OH) in the assay buffer (50 mM

NEREZ pH 7.5, 5 mm Massi", 1 mm DTT, 0.05 95 BSA). After 15 min, add 10 μl of 80 μM ATP in assay buffer. The final concentrations during the analysis were equal to 5 NM RIM'I, 40 μM ATP, 100 μM peptide VAYU and 2.595 DMSO. The reaction is carried out until approximately 50 95 ATP is consumed, then it is stopped by adding 20 μl of a solution of Kipazedio Rii5 (Rgoteda Sogrogayop). When the reaction is stopped, the mixture is incubated for 10 min and the remaining ATP is determined by luminescence using the device

Misyug2 (RegKip EITeg). The compounds of the above examples are studied by studying the consumption of ATP under the influence of RitZ and find that they are characterized by the ISvo values listed below in Table 3.

Example 143

Study of cell proliferation

KM511 cells (human myeloma lines) were grown in IMOM medium (Dulbecco's needle-modified medium) supplemented with 10 95 FBS (fetal bovine serum), sodium pyruvate and antibiotics. On the day of the study, cells in the same medium at a density of 2000 cells/well were seeded into the wells of 9x6-well tissue culture plates so that the outer wells were empty. MMTI.5 cells (human myeloma lines) were grown in KRMI1640 medium, to which 10 95 FBS, sodium pyruvate and antibiotics were added. On the day of the study, cells in the same medium at a density equal to 5000 cells/well were placed in the wells of 96b-well tissue culture plates so that the outer wells were empty.

The studied compounds obtained in DMSO were diluted in DMSO 500 times to the required final concentrations and then diluted in the culture medium 2 times to the final concentrations.

Equal volumes of 2x compounds were added to the wells of 96-well tablets and incubated at 37 "C for 3 days.

After 3 days, the tablets were equilibrated at room temperature, and the same volumes of the reagent SeiPTieg-Siom/ (Rgoteda) were added to the wells with the cultures. The tablets were briefly mixed and the luminescence signal was determined using a luminometer. Percent inhibition was calculated from the signals for cells treated with DMSO alone and cells treated with the test compound, and these values were used to determine EC values (ie, concentrations of the test compound required to produce an effect in the cells equal to 50 95 of maximum) for the studied compounds, which are listed below in Table 3.

According to the methods described in examples 140 (study of ATP consumption under the influence of Rit!), 141 (study of ATP consumption under the influence of Rit2) and 142 (study of ATP consumption under the influence of Ritud), described above, the concentrations of ICso were determined for the compounds of the above examples, which are given in table 3.

According to the methods described in example 143 (study of cell proliferation), ESvo concentrations were determined for the compounds of the above examples for KM511 cells, which are listed in Table 3.

Table C

Example Me) | Structure yet Riti Ri? (Riv KM Y identification

MKM.

N | | N r Uuye I nVttchTtinnnshnn na a t dl ny a a a o p an p p a a a i I y vinne NIM t r Chiral | | - ! She snai ("7th she in 00010018 0.006 17.6 abr shi she iz now tire kny nn i opor all, vv, 2 | | | ! . and she 7 gr ! what | s dop i V i 2 KI oh 0001 0.001 0.001 0.07

Shawn nyin shi shi i drink en sere oo shi pi p M MO

With M in Wei M0.001. 0.001 |/0o00th plo SHY ! what; and ! : nach shi tires

M ikh N: nn zh naya aa a pie en nyny nn SHY vin

And and eat. raspy! neon A, 4 I Z oooi 0003 byu 3

OO | ! ! ! and ve : : | !

N and | and shi Pone eem mi nia V ! not

E ry 0003 0.020 0009 48 deer neck N s o N

M. N

OR the real me

Not And I and 6 more g that 02 biz 000034 uyu shi

Be Y

"Example Me/ 000 Structure 00001 | Rit2 Reeves KM and , identification | ІС«ео Сб pe "De

M MUR MKM MKM MEM b.

AND

MKM

NVO early sY sya r t sho oh vv

І ше ве 0001 0.008 0.002 |033 шиШОнших ши ши ши | | to her Myrapny shi I | | And shi write srobo Zh. and and Yi and

IV | ie ra and bean (0.04 (0.02 ovi

Sh iv A N iy Eshenya y shiM pnya pp tt NN pt / pog y m o

SA o ki pi E and AK! and now, now, now, not MN, 19 g, still barefoot, ooob 29 swashes

Sh | nab Mn. SS I! ' kv gu Z ve Y E 14 N re she chiki: neck n zu v. and | these in C 0.01 0005 004 2.0 shi elI nin nn nin nn mn nin cher LA

IZ in battle 0.010 0.004 24 ! One

ST shi | | And kish ie and not NN NIC A NN nn

N gy SN Hiralnyai and | | t nok n GC, and : | Oh! ! ! !

IZ a o obi 10004 |0.003 0.67 father

M | and chik she AI

Example Mo/ | The structure of STR it! m and identification | | ISzo I iSza ISxze | "more

M MUR | MEM MKM μm | ЭСхо and мкм tt plan, mon petniatfretyutntnyaya work shsh shi

And E

114 | S ts br pov 0-4o | 0012 | 8.5! Ozh Myin N N

So shi shsh pa p V VO PI ! and oh ppyria with dralny! and Y t Y ono. Phi mn; mA, | | ! and E and N Y sha (003 0027 /olov 55 abr! ! | !

SA! | I Mon nn nn Mon Mon nn nn a she 0.001 00003 00003 117

Ii, Zk ki 16 otu gy shi shk ! | | : in renia Sh i | ! t ns rea «M (Y yabschr ! i 17 in baby 0.013 (0005 13.5 ! SG i : and ped keru KIBYLNYY ut EH, Sho

IS

18 | and Vk (0.003 0.062 000763 ! obu Ш sg y - and in E ge 7 I

And E SHY! 19 | g In her 10003 10054 0.007 14.5

YOU are writing pok sni WE o o V i vi ! l -x | and with ! '

And ZY water byu (20 ! I would 0.001 0007 0.003 1.5 ! : kak Ai su she,

PON OP KO ZAV A KON V POV VV identification and about | Mshyu | roiz | KM5Y x mur syu syu (sya vo mkm mkm mkm ALLYU shi shi she m ! sleepy: ША psa и ши -- 21 | С Ж, and

With | 0006 353

I am with her moan

F g ry | And!

Bosch 0.001 | 0.002 0.003 02

M vn nan | І Н Н poshi ny shi shin and on A and Z | and 23 | Id Ya "Y 0.002 0.005 0.003 I ra: nin o o i Yigi i ! m odini ni i i 001 0.002 Kz

A A | ; 0.001 042 whispers of controversial and | listen and listen! I kn o 001 Op ko Ku 0.001 0.002 6062 0.37 s shsh shi b it | and 26 pu i

Ve n From the battle! (0.002 0.002 ok

SYA 8 " | in E Kg | pop | 0003 0002 I suk» shi mi an aa shk she

(Example/ 00 Structure 00 bit! Level? ofRitu | KMBP identification |

M MUR | | mm μm | MEM | ES and | | | MEM now nn shin! what AI

Ve" gy | !

And E

. | tu i U bob: 10011 0000335 iy. they wash: sh a ny t sl DSN valny | | Er

But hu HA,

І 29 ше 000: 0004 0003114 ki и ми ий ВВ и НЕ s nin shi shi

Fri to ОН taking t tires now and then, 14, and E and H and | even by 0.002 | 0012 10004 zizhu

M | IJ y p v ne x cha, Y r peni t rm BU ZY e

With | in and 2, 0.002 0007 0004 |i

I am at Mor Yi she shi shi rent rt vd r | | -- a set of 32 | in would 0001 Ф004 0003 |039

В А со : А : В ! go | | ' what about vyshiny write p us: i tre Chiralny i ak i ze | No! ! ! y o y note /0.001 (05009 (0.03 4

Spirit | | ! ke | ! honors nn an i p i MON NI t NO. With Chiralny you write for | sha (0004 0067 000669

M ik | ! ; ! highway and ShSHA-.x AEYUSHYUSHSHVKA-SH ЗШ 0 5 yuyu 00001 SHI

Example Me/ | The structure of IIIrit. ory tue SHO identification | | ISkhv | ИСхв ИС | "as m MU? | | MKM | MEM MEM IES and and and MKM her

But it's crazy! y U, ve | 0.001 0.006 ooo be when M and | : not | shi write shi - write; OM Khralvnyi and : N yak S g, Tires and v shk Rada ipndya 185 36 | It is 0.001 |0.093 | 0.005 I sew shi shi shi shi shi she | he nnnyshiv: well live and All, | A. | | | and there is; | IF ve i0.002 |0.007 | 0.005 | р буры: NO I. ; I m sv) | and art

Nm SN, Y in 38 | in noc 0001 10004 0.003 1073

ХОВ ож sg ше пи тт он хХирапокий I И Й | Ave

NM sn, p 39 ni ve I a,007 (oz 001266

In their neck and pi ps V nyat tche mn, ron (40 neck and "y (ooo (0.003 0002 0.99

I was with Buli! | | | | and she she she in nn oo mouth ot

Gen i: shi i en ni yn ni 4 | - nah 0.002 10.027 0.005. re she she she she

She p sh

Example Structure of City GRita (Rishu (KM identification | ISkha ISeo ISev |-Yiche

Me MUR MEM oh mem | MEM ESzo

MEM and I and San khralnyai pi a

KOsklo. eru kad dddya Ada i, 42 she necha 0001 0.002 10002 13 shiv shi she she what nn sons shi now! nom sa: sn, s ze 43 | I and 2 (0001 0.006 |0.002 (22

M zhy: N and S pil pory on d IKVVLENY perentruettya nku, (44 vam a FObi 0.002 0002 19

And you are mine

Sushi and g i v shn p i ru s y 45 well 0001 10002 0002 /13

H in U H:

X nam 7 ---- r vederu What (46 she "y g (oo 0.002 10002 0.76 ! | Sai ; Z N ! | N su 1 ni pn i t NN van no NI

NU, AH MV : | Yah Y 47 dad 0.001 0.004 0.003 with spi and child and and

I'm going to work today, Monday, Friday, Saturday, Sunday 18 0.007 0.076 | 0.009 ! their Ky N

I am

Example Mi 00 Structure Z Ritshi |Rit? about Rishi |KM5 that G. shi identification | Yesiav IEEe6 5 | "with

Me MUR | and μm | MEM | µm | EEho mcm length of bus

And I drive in 49 and not in 0001 0.003 10002 1.8

SYA a and E ren San kirapny shin ni! not tv she bu oboi 10003 10002112 sly shi p to tt nkr y. 7 mesh sword 0001 06002 00010 51 more: in and 01 o st

Sho Sh rya ro i a nshshnshi nnai na pon ne, X ; А, 52 чо ня 0001 10.003 0003 |0.83 7 in В ови І и гу чи о ! and pi nin and nin nn that S

And Vk is (53 more "Koch (0.010 0.149 0.065 o 9 ! 4 ptnntfrentyat nannies with

Take | and shi sh for (o) in (0.003 (0.026 | 0.024 | A

YAN | It is s ny shi shi she niny nin tn ny nin nyin pnya po still ro News would | And | | ! and 55 shosh: byu 0011 3003049.

Same with 9 : : ! H na A a a PO MAV WE

An example in 7 bruyuura about and | Tweet structure: ГРый? Higher KM. identification / Shchi she ish3 CMU

M MUR So | Se S re

MEM | MEM about | µm | ESgo

Ks sh. | ! AND

SHY nin she! nm

What else Z shin nn 56 rya Iv Y doby det bovi 0y02 in schi vi oo

Ku i zh, Khirvlny penya pika 57 | In I rr ra o N ? you are 0.02 and ; sia | | shi ze: now only ni di dat nich KA | now 58 | mo, write

And! M and |. be in both battles! and: rennia What | ! ! not tt steyattrennia nkotchisn, M Ya. | shi 59 xx re H | She r Ku dobi bob because I | 001 | 9,003 0002 still on Fri nn hundredth print and | m and | | nt m | 2 sleep, m H (in still FI g

Her p bob o

And M. and |, with from 0.005 that 003 І K was in | hey! zm o! kash shi! she sword I nin beer mon gays Z T f i bod | Ii di ; ;

Hud VI February 0006 0004. 12

SHY ze ge)

Н As if in a and : by them 62. | the truth! | AH A bob1 10007 0003 |24 r g , she | | what else nn l shh san nn shshshshshshshshsh

Example Le, | The structure of OTrRini Rit?2 IRit3 | KM8P identification | 150 IS» ISv | -Ne me MUR mm o |imkMoO μm EC

AND! | | | MKM

Sh | also "A, 0001 0.003 0.002. | 0.53 more than 64 more 9.002 0.076 10005556.

M; and

N N N | ! he she: | : ! ! Company A MN; And as :

IZ s "A | | ! 65 І І 0001 0004 0093 0.16 buh and !

She about nm, | : shchi i - С с | and 66 more 0001 0004 0.002 0.52 zn o mn, i no n Kk nya obo0i 0.007 0.003 | 13 fe : and S i MM | | ! | !

I: M x shi

Z a pi z kg Y i 2 E v V g ooo ooo 0.004 14 kash shya : | B s more,

GPriklah Me; (Structure 00000 Riti o Rit?2 |Rit3 0 KM dentifation | | IS ISkhv | ISseo | -yas

MUR | | MEM about HmKM cm ES

And Sh | | | MEM and ! nak Hivaynyi And ! ; . I ! so F (9,003 0007 00061 i ! | sko Kuapkyi. Y - i

E ' will cover AK, | : tyu | X HY 0.001 0.002 | 0.002 048 nam Fi si ZE ' : : N deteu KIRAPNY : yat put SHI

Ki va Ya i (72 | game 0.095 | 0.522 0.369 daily life Kfapmny 73 Ai S 0.001 (0008 000414 chat in

N tya me i shk i i i 74 AI AA 0001 60017 10004 129 st | | what

I and cheyutya S: H 75! ver OO 0.005 0.003 12.4 sh

Examples 000 | Strukhura 0000000 tiv Rito (Rita 1KMVP identification | | KYUv ISzo | 1С50 | -Ше

M MUR | MEM | MEM | MEM | EShoa; | | MM nn PININNNNN nia oo shi 76 she a |o001 |0.00 (0002 (0.83 only

In NE village. " | U, bu (ooo! (013 (000339 nn and an To. dn miy rya

Mn, ! And ' I 78 | S hi |0.002 (0015 | v.z 3.6 govo shi shi ! she ashe | | | : si je

In | -k E H : 79 | Mr. g chi (0.002 | 0.020 (0003 pi M - Н H shi shi;

Ken ni tires shi shi what | this 000 0044 000775 veins sut shi she she soya KidVlnyi A : І t 7 Ta, vi | where (0.002 /0.025 0.005 5.940

Art N: and; ЗИ и и ге 0.003 0.0800.009 о оби дея З ! And! more

IExample Mb/ 000 Vita structure ! Mouth? | Rit3 | KM of identification and iSev so IISkhv -Me

M MUR | And µM | chkM MEM EsS:o: ! and and MEM

And I and I and Pen and nn: the same way in Mn, and e | ! And

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Example 144

Biological Techniques: Pharmacological Target Modulation and Efficacy Study for a Multiple Myeloma Xerograft Model

Cancer cells of multiple myeloma KM511-Iis, taken in Zi7appe Tgiaei (pimegeiu Neaip Meiogk,

Togopio, Sapada), to which the ability to stably express luciferase was added with the help of retroviral transfection, were stored in Golk's medium modified according to the Dulbecco method with the addition of 10 95 heat-inactivated fetal bovine serum and 1 95 glutamine (Ipmygodep,

Ips.). Female mice ZSIU/ba (age 8-12 weeks, 20-25 g, Spapev Kimeg) were used for all pharmacological studies i.p. mimo. Mice were housed in cages and handled in accordance with state and federal guidelines for the treatment of humans and laboratory animals and provided with ad libitum food and water. Cancer cells were collected from the cultures in the middle of the logarithmic growth phase, the number of viable cells was determined using equipment for automatic cell counting (Mi-SEY EI, VesKtap-SoyNeg) and the cells were resuspended in equal parts of balanced salt solution of Hanks and Maihide! (Ipmygodep, Ips.).

10 million cells were injected subcutaneously into the right side of each mouse. Compound treatment was initiated when tumor volume reached 250-350 mm3 in RK/RO studies and 150-250 mm3 in efficacy studies, and tumor volumes were determined using 5ishaubigesiog software (Eishauod Buzietv, Ips.). In treatment, all compounds were administered orally.

In the study of ip mimo modulation of the target in the case of studying the dependence of time for RK/PO, tumor-bearing mice were orally administered a single dose of diluent or compound at various concentrations.

After 1, 8 and 24 hours. after administration, tumor tissue and blood samples were taken in individual mice. Excised tumor tissues were quickly frozen and ground using a liquid nitrogen-cooled cryogenic mortar and pestle. Blood samples were collected by cardiac puncture and plasma was separated using centrifuge tubes containing lithium heparin in a plasma separator (VO Misgoiaipeg). Frozen plasma samples were lysed according to the manufacturer's instructions in a cold buffer (Meso 5saie rizsomeg) to which a protease inhibitor (Cospe) containing no ethylenediaminetetraacetic acid, phosphatase inhibitors 1 and 2 and them Mag (Zidta) was added. After homogenization in a Downs apparatus or a MaomaA GG lysing apparatus (Cospe) and after centrifugation at 300 x h for 30 min at 4 °C, a clear supernatant was obtained and the protein concentration was determined using BSA (Viogaa). Target modulation was assessed using a Meso double kit. Zsaie phospho-Vaazep!2/total Wai according to the manufacturer's instructions Briefly, the procedure was as follows: equal amounts of bileu were placed in each well of a double 96-well plate Meso Zsaie phospho-5egipe!!?7/total Wai (Meso Bsaiie Oizhsomegu) and shaking samples were incubated for 30 min at room temperature or overnight at 46.

Tablets were washed with M5O washing buffer and Z!yMo-Tad detecting antibodies were added to the wells and incubated for 1 hour with shaking. at room temperature. The tablets were washed once more and the content of the bound substance under investigation was determined after adding T reading buffer to the wells. The tablets were read using the ZESTOK Itadeg 6000 device (Meso zZsaie

Oibzsomegu). The ratio of the pVai signal to the total Va signal was used to correct the survival rate for different samples. The data presented in the table below characterize the inhibition by typical compounds proposed in this invention and represent the percentage ratio of the degree of phosphorylation of pVaidzeg!!2 to the degree of phosphorylation of the total

Defects normalized to animals of the control group, which were injected with diluent. The degree of modulation was expressed as a percentage compared to animals of the control group, which were injected with a diluent (NO, not established). 11111111 990bb0muyu) 777 77711740 | 1755 Sh | 0 71111111 .99000mu 77 | 77717162 | 66 2 | 24 11111111 7o0bOomukyuu 777777 77711728 | 762 | sh 0 g F 11111111 t7o0(ybbmuzhu 7777 77771175 | 67 | 68 gF 11111111 96o5mi)u 7 | 77717011 24 | NU 11111111 9v (bbmuyu) 77 | 7771744 | 771769 | every 16

To study the effectiveness of mice with tumors, they were randomized into groups with equivalent fluctuations in tumor volumes in the range of 150-250 mm3 using the zschmaubigesiog software (Zitsau! od Zuzietv, Ips.). After randomization, mice were orally administered the compound in 200 μl of diluent at various concentrations 1 or 2 times a day. The growth of the tumor and the body weight of the animal were determined at least 2 times a week, and a clinical examination was carried out daily to study the possible toxicity of the drug. Animals were excluded from the study if the volume of the tumor exceeded 2500 mm or the decrease in body weight exceeded 20 95 from the initial value.

The effectiveness of the compound of example 99 was evaluated using the KM511 xenograft model-

Is, when mice were orally administered the compound of example 99 twice a day at 50 and 100 mg/kg and once a day at 100 mg/kg for 14 days. Administration was started when tumor volumes were equal to approximately 250 mm3. As shown in fig. 1, the compound of example 99 shows a dose-dependent effect of ip mimo and suppression of tumor growth is observed with the introduction of 50 mg/kg twice a day (92 95) and 100 mg/kg twice a day (regression 4 95). Administration once daily at 100 mg/kg was less effective (6595) than administration twice daily. These results correlate with the degree and amount of modulation of pVaidzeg"!2 and show that significant and long-term modulation of the target is necessary to ensure maximum efficiency.

The effectiveness of the compound of example 70 was evaluated using the KM511 xenograft model

Is, when mice were orally administered the compound of example 80 twice a day at 25 and 50 mg/kg and once a day at 100 mg/kg for 14 days. Administration was started when tumor volumes were equal to approximately 225 mm3. As shown in fig. 2, the compound of example 70 shows a dose-dependent effect of ip mimo and suppression of tumor growth was observed at 25 mg/kg (65 905) and 50 mg/kg (100 95). Significant suppression of tumor growth was also observed at 100 mg/kg once daily (84 9b).

The effectiveness of the compound of example 96 was evaluated using the KM511 xenograft model

Is, when mice were orally administered the compound of example 96 twice a day at 25 and 50 mg/kg, and once a day at 100 mg/kg for 14 days. Administration was started when tumor volumes were equal to approximately 225 mm3. As shown in fig. 3, the compound of example 96 shows a dose-dependent effect of ip mimo and suppression of tumor growth is observed with the introduction of 25 mg/kg (67 905) and 50 mg/kg (96 95). Significant suppression of tumor growth was also observed at 100 mg/kg once daily (88 9b).

Although illustrative embodiments are presented and described, it should be understood that various changes may be made to it without departing from the essence and scope of the present invention.

Example 99 (oral administration) --- Diluent 700 mg/kg 2 times a day, example 989 -- 100 mg/kg 2 times a day, example 99 -U- 100 mg/kg 3 times a day, example 99 600

IF ts t 500 7

Her . same 400

M g is - is Z 7 b

Gay

Kk r 200 r) a

Phone: 0 20922 74 26 28 30 32 M

Days after implantation

Fig. 1

Example 70 (oral administration) 7000 "Ff- Diluent 900 ey 25 mgkg 2 times a day, example 70 "sh BO mg/kg 2 times a day, example 70 o 800 -9- 100 mg/kg 1 time a day, example 70 o sh- 700 g. x 500 t x-

I am in

There is

E 400

Gay a. be 300 h and shk 8 290 100 o « 17 19 21 23 25 297 29 D (3

Days after implantation

Fig. 2

Example 96 (verbal input) 10004, en. Diluent 900 -y- 25 mcg 2 times a day, example 98 -- BO mykg 2 times a day, example 96 o 8004 --100 mg/kg 1 time a day, cold 96 o y d- 700

Zh. and x 600

Ge

Зх 500 и в 400 а а и -еО 300 х ой в 200 100 о 17 19D B 23 25.27 щ293 D 3

Days after implantation

Fig. 3