EA046887B1 - ANTIBIOTIC COMPOUNDS, METHODS FOR THEIR OBTAINING, PHARMACEUTICAL COMPOSITIONS CONTAINING THEM, AND THEIR APPLICATION - Google Patents

Description

Field of technology

The present invention provides new compounds that are therapeutically active as antibiotic compounds. In addition, the present invention provides pharmaceutical compositions containing the compounds of the invention. The use of these compounds and compositions for the treatment of bacterial infections caused by N. gonorrhoeae is also presented. In addition, methods for preparing the compounds of the present invention are also provided.

BACKGROUND OF THE INVENTION

Neisseria gonorrhoeae is a type of gram-negative diplococcal bacteria responsible for the sexually transmitted infection gonorrhea, which usually causes uncomplicated (uncomplicated) disease.

The most common site of N. gonorrhoeae infection is the urogenital tract. Men with this infection may have dysuria with penile discharge, and women may have mild mucopurulent vaginal discharge and severe pelvic pain. Other infections caused by N. gonorrhoeae include anorectal, conjunctival, pharyngeal, and ovarian/uterine. In women, gonococci can cause endocervical infections or diseases of the upper genital tract. Gonococcal infections in women are closely associated with infertility. If left untreated, local N. gonorrhoeae infections may progress to bacteremia with associated septic arthritis. When the infection affects the eyes, especially in newborns, blindness can occur if not treated promptly. In rare cases, bacteremia leads to spread (complicated gonorrhea) to the joints, skin, endocardium or meninges.

Treatment of gonorrhea is complicated by the ability of N. gonorrhoeae to develop antimicrobial resistance. In 2007, the emergence of fluoroquinolone-resistant N. gonorrhoeae in the United States prompted health authorities to stop recommending fluoroquinolones for the treatment of gonorrhea in the United States. Recent treatment failures with cefixime or other oral cephalosporins have been reported in Asia, Europe, South Africa and Canada. Failures in the treatment of pharyngeal infections with ceftriaxone have been reported in Australia, Japan and Europe. Consequently, global organizations such as the WHO recommend that local resistance data guide the choice of therapy.

Recently, there have been isolated reports of N. gonorrhoeae strains resistant to both cephalosporins and macrolides, which leaves no treatment options for these patients. There are concerns that due to the ability of Neisseria sp. By rapidly spreading and integrating their genetic material (promiscuous transformation), these hyper-drug-resistant genotypes can spread rapidly.

FabI inhibition is a relatively new concept for antibiotic development. FabI is an enzyme involved in the synthesis of bacterial fatty acids. Administration of compounds that inhibit this enzyme can selectively target bacteria dependent on this enzyme without affecting the host (patient). Antibiotic agents based on this principle of action are described, for example, in WO 03/088897 A2 and WO 2013/190384 A1, the contents of which are fully included in the present application. However, these compounds are primarily intended for the treatment of infections caused by S. aureus. Other compounds with an inhibitory effect on FabI, but aimed primarily at the treatment of infections other than N. gonorrhoeae, are disclosed in WO 2007/053131 A, WO 2007/067416 A, WO 01/27103 A, WO 2008/098374 A, Yao et al., J. Biol. Chem., 2016, 291, 171-181, WO 2008/009122 A, US 2015/210719 A and US 2017/174683 A.

Problems underlying the invention

In view of the above situation, there is a need to provide additional drugs and pharmaceutical compositions that exhibit antibiotic activity against N. gonorrhoeae.

It is particularly desirable to provide such other drugs and pharmaceutical compositions that can be used as an alternative treatment for bacterial infections caused by N. gonorrhoeae when standard treatment with ceftriaxone is ineffective.

In addition, there is a need for additional antibiotic drugs and medications to treat various related bacterial infections.

In addition, it is desirable that such antibiotic drugs and pharmaceutical compositions exhibit an acceptable therapeutic index and do not have serious side effects.

The essence of the invention

The present invention solves the above problems by providing compounds and pharmaceutical compositions that are effective in treating infections caused by N. gonorrhoeae and related bacteria.

The compounds of the present invention act as antibiotic agents through a working mechanism involving FabI inhibition. This mechanism differs from that of cephalosporins, which act through the destruction of peptidoglycan layers of bacterial cell walls. Moreover, the compounds of the present invention have a chemical structure different from cephalosporus

- 1 046887 new Therefore, it can be expected that the compounds and pharmaceutical compositions of the present invention are also effective against strains resistant to ceftriaxone.

The compounds and pharmaceutical compositions of the present invention further show a high therapeutic index and low side effects.

In one embodiment, the compounds and pharmaceutical compositions of the present invention exhibit chemical, physical and pharmacokinetic properties suitable for oral and/or intramuscular administration.

In one embodiment, the present invention also provides therapeutic uses of the compounds described herein and methods for treating bacterial infections comprising administering the compounds of the present invention to patients in need thereof.

In other embodiments, the present invention also provides methods for preparing the compounds of the present invention.

Thus, the present invention provides the following embodiments/

Embodiment 1. A compound that is selected from the group of compounds represented by the general formula I where A1 represents a group selected from the groups A _n and A ₁₂ having the following structures: H ₃ with where the line connected to the exocyclic methylene group represents a single covalent bond formed with a nitrogen atom of formula I;

A2 represents a methyl group;

or A1 and A2 together with the nitrogen atom to which they are bonded form the following group A3:

where the exocyclic line connected to the nitrogen atom of the bicycle represents a covalent bond between the nitrogen and the carbonyl group on the left side of formula I;

and where in option (iii)

Q1 represents CH2 or NH;

Q2 represents CR ⁴ R ⁵ -CR ⁶ R ⁷ where the CR4R ⁵ group binds to Q1;

or Q1-Q2 is -N=CR ⁷ -CHR ⁷ - or its tautomer -NH-CR ⁷ =CR ⁷ -;

Q3 represents O or S;

R ¹ represents a group selected from H, -NH ₂ ;

R ² represents a group selected from H, -O-Ar ¹ , -O-Het ¹ , -NR ⁹ R ¹⁰ , -O-Alk ¹ , where Ar ¹ represents a phenyl group which may be optionally substituted by one or more groups, selected from -CN, -O-Ci-4-alkyl, -O-(CH ₂ ) _1-4 -NR ⁹ R ¹⁰ , or wherein the phenyl group may bear two substituent groups on adjacent ring atoms such that the adjacent substituent groups can be linked together to form a 5-membered heterocycle having one or two heteroatoms individually selected from N and O, where Het ¹ is an aromatic heterocycle with 5 or 6 ring atoms comprising 1 or 2 heteroatoms individually selected from N, S and O, or non-aromatic partially or fully saturated heterocycle with 6 ring atoms containing 1 heteroatom, you

- 2 046887 brane of N and O, wherein the Het ¹ group is optionally substituted by one or more groups individually selected from -^^-alkyl, -OQ^-alkyl, -CN, -(CH2) _0-4 -OH; where R ⁹ is selected from H and -C.'i-.-1-alkyl; where R ¹⁰ is selected from H, -Ci i-alkyl and -C(=O)-CH3; where Alk ¹ represents an alkyl group having from 1 to 6 carbon atoms, which is linear, branched or cyclic, or a combination thereof, where the Alk ¹ group is optionally substituted by one or more groups selected from -OH, -O-alkyl;

R3 represents a group selected from H, -PO3R ^3a 2, -CH2-OPO ₃ R ^3a 2 and -CH2-OC(=O)-R ^3b ;

R ^3a represents a hydrogen atom or cation suitable to form a pharmaceutically acceptable salt of either -CH2-OC(=O)-R ^3b or -CHMe-OC(=O)-R ^3b or -CMe2-OC(=O)- R ^3b , or -CH2-OC(=O)OR ^3b , or -CHMe-OC(=O)-OR ^3b , or -CMe2-OC(=O)-OR ^3b ;

R ^3b represents an alkyl group having from 1 to 11 carbon atoms, which is linear, branched or cyclic, or a combination thereof, wherein the R ^3b group may optionally be substituted by one or more groups independently selected from -OH and -OC _1-6 -alkyl;

R ⁴ represents a group selected from H, ^^-alkyl, CN and C _1-4 -alkylene-F;

R ⁵ represents a group selected from H, ^^-alkyl, C _1-4 -alkylene-OH, C _1-4 -alkylene-OR ³ , -OH, -OPO3R ^3a 2;

or R ⁴ and R ⁵ together form a cyclic group having from 4 to 6 ring members formed by methylene groups and, optionally, an oxygen atom;

R ⁶ represents a group selected from -OH, C _1-4 alkyl, -OPO ₃ R ^3a 2;

R ⁷ represents a group selected from H, C1-4-alkyl, C1-4-alkylene-OH, C1-4-alkylene-OR ³ , C1-4-alkylene-F, -CN;

or R ⁶ and R ⁷ together form a cyclic group having from 4 to 6 ring members formed by methylene groups and, optionally, an oxygen atom; wherein the cyclic group may optionally bear a substituent selected from -OH, -O-alkyl;

R ⁸ represents a group selected from -O-Ar ² and -O-Het ² ;

where Ar ² represents a phenyl group which is optionally substituted by one or more groups selected from -CN, -OC _1-4 -alkyl, or where the phenyl group may bear two substituent groups on adjacent ring atoms such that these adjacent substituent groups may be bonded together to form a 5-membered heterocycle having one or two heteroatoms individually selected from N, S and O, wherein said heterocycle may optionally bear one or two substituents selected from oxo, halogen, -OC _1-4 -alkyl, ^ ^-alkyl and CN; and where Het ² represents an aromatic heterocycle with 5 or 6 ring atoms, including 1 or 2 heteroatoms individually selected from N, S and O, which may optionally be substituted by one or more groups individually selected from -OC _1-4 -alkyl, -CN, F, C _1-4 -alkyl, and pharmaceutically acceptable salts thereof; or in option (iv)

Q ¹ represents CH2 or NH;

Q2 represents CR ⁴ R ⁵ -CR ⁶ R ⁷ where the CR4R ⁵ group binds to Q1;

or Q1-Q2 is -N=CR ⁷ -CHR ⁷ - or its tautomer -NH-CR ⁷ =CR ⁷ -;

Q ₃ represents O or S;

R ¹ represents a group selected from H, -NH ₂ ;

R ² represents a group selected from H, -O-Ar ¹ , -O-Het ¹ , -NR ⁹ R ¹⁰ , -O-Alk ¹ where Ar ¹ represents a phenyl group which is optionally substituted by one or more groups selected from CN, -O-C1-4-alkyl, -O-(CH2)i-4-NR ⁹ R ¹⁹ , or where the phenyl group can bear two substituent groups on adjacent ring atoms so that these adjacent substituent groups can be bonded together to form a 5-membered heterocycle having one or two heteroatoms individually selected from N and O, where Het ¹ is an aromatic heterocycle with 5 or 6 ring atoms containing 1 or 2 heteroatoms individually selected from N, S and O, or not an aromatic partially or fully saturated heterocycle with 6 ring atoms including 1 heteroatom selected from N and O, wherein the Het ¹ group may optionally be substituted by one or more groups individually selected from -CY.-i-alkyl. -O-C1-4-alkula, -CN, -(CH ₂ ) _0-4 -OH; where R ⁹ is selected from H and -CY.-i-alkyl; where R ¹⁰ is selected from H, -^^-alkyl and -C(=O)-CH ₃ ; where Alk ¹ represents an alkyl group having from 1 to 6 carbon atoms, which is linear, branched or cyclic, or a combination thereof, where the Alk ¹ group may be optionally substituted by one or more groups selected from -OH, -O-alkyl;

R ³ represents a group selected from H, -PO3R ^3a 2, -CH2-OPO3R ^3a 2 and -CH2-OC(=O)-R ^3b ;

R ^3a represents a hydrogen atom or cation suitable to form a pharmaceutically acceptable salt of either -CH2-OC(=O)-R ^3b or -CHMe-OC(=O)-R ^3b or -CMe2-OC(=O)-R ^3b or -CH2-OC(=O)-OR ^3b or -CHMe-OC(=O)-OR ^3b or -CMe2-OC(=O)-OR ^3b ;

R ^3b represents an alkyl group having from 1 to 11 carbon atoms, which is linear, branched or cyclic, or a combination thereof, wherein the R ^3b group is optionally substituted by one or more groups independently selected from -OH and -OC _1-6 -alkyl ;

- 3 046887

R ⁴ represents a group selected from H, C _1-4 -alkyl, CN and C|—.|-alkylene-H;

R ⁵ represents a group selected from H, CM-alkyl, C1-4-alkylene-OH, C1-4-alkylene-OR ³ , -OH, -OPO3R ^3a 2;

or R ⁴ and R ⁵ together form a cyclic group having from 4 to 6 ring members formed by methylene groups and, optionally, an oxygen atom;

R ⁶ represents a group selected from H, -OH, N-alkyl, -OPO ₃ R ^3a 2;

R ⁷ represents a group selected from N-alkyl, C1-4-alkylene-OH, C1-4-alkylene-OR ³ , C1-4-alkylene-F, -CN;

or R ⁶ and R ⁷ together form a cyclic group having from 4 to 6 ring members formed by methylene groups and, optionally, an oxygen atom; wherein the cyclic group may optionally bear a substituent selected from -OH, -O-alkyl;

R ⁸ represents a group selected from -O-Ar ² and -O-Het ² ;

where Ar ² represents a phenyl group which is optionally substituted by one or more groups selected from -CN, -OC _1-4 -alkyl, or where the phenyl group may bear two substituent groups on adjacent ring atoms such that these adjacent substituent groups may be bonded together to form a 5-membered heterocycle having one or two heteroatoms individually selected from N, S and O, wherein said heterocycle may optionally bear one or two substituents selected from oxo, halogen, -OC _1-4 -αalkyl, ^ c-alkyl and CN;

where Het ² represents an aromatic heterocycle with 5 or 6 ring atoms, comprising 1 or 2 heteroatoms individually selected from N, S and O, which may optionally be substituted by one or more groups individually selected from -O-CM-alkyl, -CN , F, CM-alkyl, and its pharmaceutically acceptable salts, where the term alkyl refers to a saturated straight or branched hydrocarbon of 1-8 carbon atoms;

a cation suitable to form a pharmaceutically acceptable salt is Na, K, Mg, Ca, or the protonated form of the organic base ethanolamine, meglumine, tromethamine (2-amino-2-(hydroxymethyl)propane-1,3-diol) or deanol (2 -(dimethylamino)ethanol).

Embodiment 2. The compound of Embodiment 1, wherein the term alkyl refers to a saturated straight or branched hydrocarbon of 1-6 carbon atoms

Embodiment 3. The compound of Embodiment 1, option (iii) or (iv), wherein R ⁴ represents a group selected from ^alkyl, CN and ^t-alkylene^; And

R ⁵ represents a group selected from H, N-alkyl, C _1-4 -αalkylene-OH, C _1-4 -alkylene-OR ³ , -OH, -OPO3R ^3a 2;

and the remaining variable groups are as defined in paragraph 1, option (iii) or (iv).

Embodiment 4. The compound of Embodiment 1, option (iii) or (iv), wherein R ⁴ represents a group selected from H, Ci-4-alkyl, CN and Ci-4-alkylene-P; And

R ⁵ represents a group selected from CM-alkyl, C _1-4 -alkylene-OH, C _1-4 -alkylene-OR ³ , -OH, -OPO3R ^3a 2;

and the remaining variable groups are as defined in paragraph 1, option (iii) or (iv).

Embodiment 5. The compound of Embodiment 1, 2 or 3, 4, wherein the compound is selected from the group of compounds represented by the general formula II

where R ¹ , R ² , R ³ , Q ₁ , Q ₂ , Q ₃ have the same meanings as defined in paragraphs 1, 2 or 3 above;

and wherein the five-membered heterocycle containing Q3 is bonded to a methyleneamide group at position 2 and a methyl group at position 3, or is bonded to a methyleneamide group at position 3 and a methyl group at position 2.

Embodiment 6. The compound of Embodiment 5, wherein the compound is selected from the group of compounds represented by the general formula III

^R III where R ¹ , R ² , R ³ , Q ₁ , Q ₂ , Q ₃ have the same meanings as defined in paragraphs 1, 2 or 3 above.

- 4 046887

Embodiment 7. The compound of Embodiment 6, wherein the compound is selected from the group of compounds represented by the general formula VI

where R ¹ , R ² , R ³ , R ⁴ , R ⁵ , R ⁶ and R ⁷ have the same meanings as defined in option (iii) or (iv) according to paragraphs. 1, 2 or 3 above, where in option (iii) it is preferable

R ¹ represents H, NH ₂ ;

R ² represents H;

R ³ represents a group selected from H, -PO3R ^3a 2, -CH ₂ -OPO ₃ R ^3a ₂ and -CH ₂ -OC(=O)-R ^3b ;

R ^3a represents a hydrogen atom or cation suitable to form a pharmaceutically acceptable salt of either -CH2-OC(=O)-R ^3b or -CHMe-OC(=O)-R ^3b or -CMe2-OC(=O)-R ^3b or -CH ₂ -OC(=O)-OR ^3b or -CHMe-OC(=O)-OR ^3b or -CMe2-OC(=O)-OR ^3b ;

R ^3b represents an alkyl group having from 1 to 11 carbon atoms, which is linear, branched or cyclic, or a combination thereof, wherein the R ^3b group is optionally substituted by one or more groups independently selected from -OH and -O-C _1-6 -alkyl;

R ⁴ represents H, C| _-4 -iuiKH.T and

R ⁵ represents H, C _1-4 -alkyl, C _1-4 -alkylene-OR ³ ;

R ⁶ represents C1-4-alkyl, OH, -OPO3R ^3a 2; And

R ⁷ represents H, C1-4-alkyl, Cy^alkylene-O^ C1-4-alkylene-OR ³ , where R ³ is as defined above, C1-4-alkylene-H; and where in option (iv) it is preferable

R ¹ represents H, NH ₂ ;

R ² represents H;

R ³ represents a group selected from H, -PO3R ^3a 2, -CH ₂ -OPO ₃ R ^3a ₂ and -CH ₂ -OC(=O)-R ^3b ;

R ^3a represents a hydrogen atom or cation suitable to form a pharmaceutically acceptable salt of either -CH2-OC(=O)-R ^3b or -CHMe-OC(=O)-R ^3b or -CMe2-OC(=O)-R ^3b or -CH2-OC(=O)-OR ^3b or -CHMe-OC(=O)-OR ^3b or -CMe2-OC(=O)-OR ^3b ;

R ^3b represents an alkyl group having from 1 to 11 carbon atoms, which is linear, branched or cyclic, or a combination thereof, wherein the R ^3b group is optionally substituted by one or more groups independently selected from -OH and -O-C _1-6 -alkyl;

R ⁴ represents H, C _1-4 alkyl; And

R ⁵ represents H, C _1-4 -alkyl, C _1-4 -alkylene-OR ³ ;

R ⁶ represents H, C1-4-alkyl, OH, -OPO3R ^3a 2; And

R ⁷ is C1-4-alkyl, C1-4-alkylene-OH, C1-4-alkylene-OR ³ , where R ³ is as defined above, C1-4-alkylene-G.

Embodiment 8. The compound of Embodiment 6, wherein the compound is selected from the group of compounds represented by the general formula VII

R VII where R ¹ , R ² , R ³ , R ⁶ and R ⁷ have the same meanings as defined in option (iii) or (iv) according to claims 1, 2 or 3 above, where in option (iii) it is preferable

R ¹ represents H, NH2;

R ² represents H;

R ³ represents a group selected from H, -PO3R ^3a 2, -CH ₂ -OPO ₃ R ^3a ₂ and -CH ₂ -OC(=O)-R ^3b ;

R ^3a represents a hydrogen atom or cation suitable to form a pharmaceutically acceptable salt of either -CH2-OC(=O)-R ^3b or -CHMe-OC(=O)-R ^3b or -CMe2-OC(=O)-R ^3b or -CH2-OC(=O)-OR ^3b or -CHMe-OC(=O)-OR ^3b or -CMe2-OC(=O)-OR ^3b ;

R ^3b represents an alkyl group having from 1 to 11 carbon atoms, which is linear, branched or cyclic, or a combination thereof, wherein the R ^3b group is optionally substituted by one or more groups independently selected from -OH and -O-C _1-6 -alkyl;

R ⁴ represents H, C _1-4 alkyl; And

- 5 046887

R ⁵ represents H, C _1-4 -alkyl, C _1-4 -aTKnTeH-OR ³ ;

R ⁶ represents C _1-4 -alkyl, OH, -OPO ₃ R ^3a 2; And

R ⁷ is H, C1-4-alkyl, C1-4-alkylene-OH, C1-4-alkylene-OR ³ , where R ³ is as defined above, C1-4-alkylene-F; and where in option (iv) it is preferable

R ¹ represents H, NH2;

R ² represents H;

R ³ represents a group selected from H, -PO3R ^3a 2, -CH2-OPO3R ^3a 2 and -CH2-OC(=O)-R ^3b ;

R ^3a represents a hydrogen atom or cation suitable to form a pharmaceutically acceptable salt of either -CH2-OC(=O)-R ^3b or -CHMe-OC(=O)-R ^3b or -CMe2-OC(=O)-R ^3b or -CH ₂ -OC(=O)-OR ^3b or -CHMe-OC(=O)-OR ^3b or -CMe2-OC(=O)-OR ^3b ;

R ^3b represents an alkyl group having from 1 to 11 carbon atoms, which is linear, branched or cyclic, or a combination thereof, wherein the R ^3b group is optionally substituted by one or more groups independently selected from -OH and -OC _1-6 -αalkyl ;

R ⁴ represents H, C _1-4 -αalkyl; And

R ⁵ represents H, C _1-4 -alkyl, C _1-4 -alkylene-OR ³ ;

R ⁶ represents H, C _1-4 -alkyl, OH, -OPO ₃ R ^3a 2; And

R ⁷ is C1-4-alkyl, C1-4-alkylene-OH, C1-4-alkylene-OR ³ , where R ³ is as defined above, C1-4-alkylene-F.

Embodiment 9. The compound of Embodiment 5, wherein the compound is selected from the group of compounds represented by the general formula VIII

where R ¹ , R ² , R ³ , Q1, Q2, Q ₃ have the same meanings as defined in paragraphs 1, 2 or 3 above.

Embodiment 10. The compound of Embodiment 9, wherein the compound is selected from the group of compounds represented by the general formula XI

where R ¹ , R ² , R ³ , R ⁴ , R ⁵ , R ⁶ and R ⁷ have the same meanings as defined in paragraphs 1, 2 or 3 above, where in option (iii) it is preferable

R ¹ represents H;

R ² represents a group selected from H, -O-Ar ¹ , -O-Het ¹ , -NH2, where Ar ¹ represents a phenyl group which is optionally substituted by one or more groups selected from -CN, -O-C1-4 -alkyl, or where the phenyl group may bear two substituent groups on adjacent ring atoms such that these adjacent substituent groups can be bonded together to form a 5-membered heterocycle having one or two heteroatoms individually selected from N and O, where Het ¹ represents an aromatic heterocycle with 5 or 6 ring atoms including 1 or 2 heteroatoms individually selected from N, S and O, or a non-aromatic partially or fully saturated heterocycle with 6 ring atoms including 1 heteroatom selected from N, S and O, wherein the Het ¹ group may optionally be substituted by one or more groups individually selected from ^^-alkyl, -O-C1-4-alkyl, -CN;

R ³ represents a group selected from H, -PO3R ^3a 2, -CH ₂ -OPO ₃ R ^3a ₂ and -CH ₂ -OC(=O)-R ^3b ;

R ^3a represents a hydrogen atom or cation suitable to form a pharmaceutically acceptable salt of either -CH2-OC(=O)-R ^3b or -CHMe-OC(=O)-R ^3b or -CMe2-OC(=O)-R ^3b or -CH2-OC(=O)-OR ^3b or -CHMe-OC(=O)-OR ^3b or -CMe2-OC(=O)-OR ^3b ;

R ^3b represents an alkyl group having from 1 to 11 carbon atoms, which is linear, branched or cyclic, or a combination thereof, wherein the R ^3b group may optionally be substituted by one or more groups independently selected from -OH and ^^^-alkyl ;

R ⁴ represents H, C _1-4 -alkyl; And

R ⁵ represents H, C _1-4 -alkyl, C _1-4 -alkylene-OR ³ .

R ⁶ represents C1-4-alkyl, OH, -OPO3R ^3a 2; And

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R ⁷ represents H, C1-4-alkyl, C1-4-alkylene-OH, C|-.|-a.iKu.ieii-(')R3, where R ³ is as defined above, C1-4-alkylene ^;

where in option (iv) it is preferable

R ^| represents H;

R ² represents a group selected from H, -O-Ar ¹ , -O-Het ¹ , -NH2, where Ar ¹ represents a phenyl group which is optionally substituted by one or more groups selected from -CN, -O-C1-4 -alkyl, or where the phenyl group may bear two substituent groups on adjacent ring atoms such that these adjacent substituent groups can be bonded together to form a 5-membered heterocycle having one or two heteroatoms individually selected from N and O, where Het ¹ represents an aromatic heterocycle with 5 or 6 ring atoms including 1 or 2 heteroatoms individually selected from N, S and O, or a non-aromatic partially or fully saturated heterocycle with 6 ring atoms including 1 heteroatom selected from N, S and O, wherein the Het ¹ group may optionally be substituted by one or more groups individually selected from -C _1-4 -alkyl, -O-C _1-4 -alkyl, -CN;

R ³ represents a group selected from H, -PO3R ^3a 2, -CH ₂ -OPO ₃ R ^3a ₂ and -CH ₂ -OC(=O)-R ^3b ;

R ^3a represents a hydrogen atom or cation suitable to form a pharmaceutically acceptable salt of either -CH2-OC(=O)-R ^3b or -CHMe-OC(=O)-R ^3b or -CMe2-OC(=O)-R ^3b or -CH ₂ -OC(=O)-OR ^3b or -CHMe-OC(=O)-OR ^3b or -CMe2-OC(=O)-OR ^3b ;

R ^3b represents an alkyl group having from 1 to 11 carbon atoms, which is linear, branched or cyclic, or a combination thereof, wherein the R ^3b group is optionally substituted by one or more groups independently selected from -OH and -OC _1-6 -alkyl ;

R ⁴ represents H, C _1-4 -alkyl; And

R ⁵ represents H, C _1-4 -alkyl, C _1-4 -alkylene-OR ³ .

R ⁶ represents H, C1-4-alkyl, OH, -OPO3R ^3a 2; And

R ⁷ is C1-4-alkyl, C1-4-alkylene-OH, C1-4-alkylene-OR ³ , where R ³ is as defined above, C1-4-alkylene-F.

Embodiment 11. The compound of Embodiment 9, wherein the compound is selected from the group of compounds represented by the general formula XII

R ¹

^to XII where R ¹ , R ² , R ³ , R ⁶ and R ⁷ have the same meanings as defined in paragraphs. 1, 2 or 3 above, where in option (iii) it is preferable

R ¹ represents H;

R ² represents a group selected from H, -O-Ar ¹ , -O-Het ¹ , -NH2, where Ar ¹ represents a phenyl group which is optionally substituted by one or more groups selected from -CN, -O-C1-4 -alkyl, or where the phenyl group may bear two substituent groups on adjacent ring atoms such that these adjacent substituent groups can be bonded together to form a 5-membered heterocycle having one or two heteroatoms individually selected from N and O, where Het ¹ represents an aromatic heterocycle with 5 or 6 ring atoms including 1 or 2 heteroatoms individually selected from N, S and O, or a non-aromatic partially or fully saturated heterocycle with 6 ring atoms including 1 heteroatom selected from N, S and O, wherein the Het ¹ group is optionally substituted with one or more groups individually selected from ^^-alkyl, -OC _1-4 -alkyl, -CN;

R ³ represents a group selected from H, -PO3R ^3a 2, -CH2-OPO ₃ R ^3a ₂ and -CH ₂ -OC(=O)-R ^3b ;

R ^3a represents a hydrogen atom or cation suitable to form a pharmaceutically acceptable salt of either -CH2-OC(=O)-R ^3b or -CHMe-OC(=O)-R ^3b or -CMe2-OC(=O)-R ^3b or -CH2-OC(=O)-OR ^3b or -CHMe-OC(=O)-OR ^3b or -CMe2-OC(=O)-OR ^3b ;

R ^3b represents an alkyl group having from 1 to 11 carbon atoms, which is linear, branched or cyclic, or a combination thereof, wherein the R ^3b group is optionally substituted by one or more groups independently selected from -OH and -OC _1-6 -alkyl ;

R ⁴ represents H, C _1-4 -alkyl; And

R ⁵ represents H, C _1-4 -alkyl, C _1-4 -alkylene-OR ³ ;

R ⁶ represents C1-4-alkyl, OH, -OPO3R ^3a 2; And

R ⁷ is H, C1-4-alkyl, C1-4-alkylene-OH, C1-4-alkylene-OR ³ , where R ³ is as defined above, C1-4-alkylene-F;

- 7 046887 where in option (iv) it is preferable

R ¹ represents H;

R ² represents a group selected from H, -O-Ar ¹ , -O-Het ¹ , -NH2, where Ar ¹ represents a phenyl group which may be optionally substituted by one or more groups selected from -CN, -O-C1 -4-alkyl, or where the phenyl group may bear two substituent groups on adjacent ring atoms such that these adjacent substituent groups can be bonded together to form a 5-membered heterocycle having one or two heteroatoms individually selected from N and O, where Het ¹ is an aromatic heterocycle with 5 or 6 ring atoms comprising 1 or 2 heteroatoms individually selected from N, S and O, or a non-aromatic partially or fully saturated heterocycle with 6 ring atoms containing 1 heteroatom selected from N, S and O, wherein the Het ¹ group is optionally substituted with one or more groups individually selected from -C _1-4 -alkyl, -OC _1-4 -alkyl, -CN;

R ³ represents a group selected from H, -PO3R ^3a 2, -CH ₂ -OPO ₃ R ^3a ₂ and -CH ₂ -OC(=O)-R ^3b ;

R ^3a represents a hydrogen atom or cation suitable to form a pharmaceutically acceptable salt of either -CH2-OC(=O)-R ^3b or -CHMe-OC(=O)-R ^3b or -CMe2-OC(=O)-R ^3b or -CH ₂ -OC(=O)-OR ^3b or -CHMe-OC(=O)-OR ^3b or -CMe2-OC(=O)-OR ^3b ;

R ^3b represents an alkyl group having from 1 to 11 carbon atoms, which is linear, branched or cyclic, or a combination thereof, wherein the R ^3b group is optionally substituted by one or more groups independently selected from -OH and -OC _1-6 -αalkyl ;

R ⁴ represents H,^^-alkyl; And

R ⁵ represents H,^^-alkyl, C _1-4 -alkylene-OR ³ ;

R ⁶ represents H, ^^-alkyl, OH, -OPO3R ^3a 2; And

R ⁷ represents ^^-alkyl, C1-4-αalkylene-OH, C1-4-alkylene-OR ³ , where R ³ is as defined above, C1-4-αalkylene-F.

Embodiment 12. The compound of Embodiment 1, wherein the compound is selected from the group of compounds represented by the general formula XIII

where R ³ , R ⁸ , Q1, Q2 have the same meanings as defined in paragraphs 1, 2 or 3 above.

Embodiment 13. The compound of Embodiment 12, wherein the compound is selected from the group of compounds represented by the general formula XV

where R ³ , R ⁴ , R ⁵ , R ⁶ , R ⁷ , R ⁸ have the same meanings as defined in paragraphs 1, 2 or 3 above, where in option (iii) it is preferable

R ³ represents a group selected from H, -PO3R ^3a 2, -CH2-OPO3R ^3a 2 and -CH2-OC(=O)-R ^3b ;

R ^3a represents a hydrogen atom or cation suitable to form a pharmaceutically acceptable salt of either -CH2-OC(=O)-R ^3b or -CHMe-OC(=O)-R ^3b or -CMe2-OC(=O)-R ^3b or -CH2-OC(=O)-OR ^3b or -CHMe-OC(=O)-OR ^3b or -CMe2-OC(=O)-OR ^3b ;

R ^3b represents an alkyl group having from 1 to 11 carbon atoms, which is linear, branched or cyclic, or a combination thereof, wherein the R ^3b group is optionally substituted by one or more groups independently selected from -OH and -OC _1-6 -alkyl ;

R ⁴ represents H,^^-alkyl; And

R ⁵ represents H,^^-alkyl, C _1-4 -alkylene-OR ³ ;

R ⁶ represents Q-4-alkyl, OH, -OPO3R ^3a 2; And

R ⁷ represents H, C^-alkyl, C1-4-alkylene-OH, C1-4-alkylene-OR ³ , where R ³ is as defined above, C1-4-alkylene-F; And

R ⁸ represents a group selected from -O-Ar ² and -O-Het ² where Ar ² represents a phenyl group which is optionally substituted by one or more

- 8 046887 groups selected from -CN, -O-C1. ₄ -alkyl, or where the phenyl group may bear two substituent groups on adjacent ring atoms so that these adjacent substituent groups can be bonded together to form a 5-membered heterocycle having one or two heteroatoms individually selected from N, S and O, where said heterocycle may optionally bear one or two substituents selected from oxo, F, -OC ₁ . ₄ -αalkyl, C ₁ . ₄ -αalkyl, CN; and where Het ² is an aromatic heterocycle with 5 or 6 ring atoms comprising 1 or 2 heteroatoms individually selected from N, S and O, which are optionally substituted by one or more groups individually selected from -OC ₁ . ₄ -αalkyl, -CN, F, C ₁ . ₄ -αalkyl;

where in option (iv) it is preferable

R ³ represents a group selected from H, -PO3R ^3a 2, -CH ₂ -OPO ₃ R ^3a ₂ and -CH ₂ -OC(=O)-R ^3b ;

R ^3a represents a hydrogen atom or cation suitable to form a pharmaceutically acceptable salt of either -CH2-OC(=O)-R ^3b or -CHMe-OC(=O)-R ^3b or -CMe2-OC(=O)-R ^3b or -CH ₂ -OC(=O)-OR ^3b or -CHMe-OC(=O)-OR ^3b or -CMe2-OC(=O)-OR ^3b ;

R ^3b represents an alkyl group having from 1 to 11 carbon atoms, which is linear, branched or cyclic, or a combination thereof, wherein the R ^3b group is optionally substituted by one or more groups independently selected from -OH and -OC _1-6 -αalkyl ;

R ⁴ represents H, C _1-4 -αalkyl; And

R ⁵ represents H, C _1-4 -alkyl, C _1-4 -alkylene-OR ³ ;

R ⁶ represents H, C1-4-alkyl, OH, -OPO3R ^3a 2; And

R ⁷ is C1-4-alkyl, C1-4-alkylene-OH, C1-4-alkylene-OR ³ where R ³ is as defined above, C1-4-alkylene-F; And

R ⁸ represents a group selected from -O-Ar ² and -O-Het ² , where Ar ² represents a phenyl group that is optionally substituted by one or more groups selected from -CN, -OC _1-4 -alkyl, or where phenyl group may bear two substituent groups on adjacent ring atoms such that these adjacent substituent groups may be bonded together to form a 5-membered heterocycle having one or two heteroatoms individually selected from N, S and O, wherein said heterocycle may optionally bear one or two substituents selected from oxo, F, -OC _1-4 -αalkyl, C _1-4 -αalkyl, CN; and where Het ² represents an aromatic heterocycle with 5 or 6 ring atoms, including 1 or 2 heteroatoms individually selected from N, S and O, which may optionally be substituted by one or more groups individually selected from -OC _1-4 -alkyl, -CN, F, C _1-4 -alkyl.

Embodiment 14. The compound of Embodiment 12, wherein the compound is selected from the group of compounds represented by the general formula XVI

where R ³ , R ⁴ , R ⁵ , R ⁶ , R ⁷ , R ⁸ have the same meanings as defined in paragraphs 1, 2 or 3 above, where in option (iii) it is preferable

R ³ represents a group selected from H, -PO3R ^3a 2, -CH ₂ -OPC _1-4 ^3a ₂ and -CH ₂ -OC(=O)-R ^3b ;

R ^3a represents a hydrogen atom or cation suitable to form a pharmaceutically acceptable salt of either -CH2-OC(=O)-R ^3b or -CHMe-OC(=O)-R ^3b or -CMe2-OC(=O)-R ^3b or -CH ₂ -OC(=O)-OR ^3b or -CHMe-OC(=O)-OR ^3b or -CMe2-OC(=O)-OR ^3b ;

R ^3b represents an alkyl group having from 1 to 11 carbon atoms, which is linear, branched or cyclic, or a combination thereof, wherein the R ^3b group is optionally substituted by one or more groups independently selected from -OH and -OC _1-6 -alkyl ;

R ⁴ represents H, C _1-4 alkyl; And

R ⁵ represents H, C _1-4 -alkyl, C _1-4 -alkylene-OR ³ ;

R ⁶ represents C1-4-alkyl, OH, -OPO3R ^3a 2; And

R ⁷ is H, C1-4-alkyl, C1-4-alkylene-OH, C1-4-alkylene- ^OR3 , where ^R3 is as defined above, C1-4-alkylene^;

R ⁸ represents a group selected from -O-Ar ² and -O-Het ² , where Ar ² represents a phenyl group which may be optionally substituted by one or more groups selected from -CN, -OC _1-4 -alkyl, or wherein the phenyl group may bear two substituent groups on adjacent ring atoms such that these adjacent substituent groups may be bonded together to form a 5-membered heterocycle having one or two heteroatoms selected from N, S and O, wherein said heterocycle may optionally bear one or two substituents, indi

- 9 046887 specifically selected from oxo, F, -O-C _1-4 -alkyl, C _1-4 -alkyl, CN; and where Het ² represents an aromatic heterocycle with 5 or 6 ring atoms, including 1 or 2 heteroatoms individually selected from N, S and O, which are optionally substituted by one or more groups individually selected from -OC _1-4 -alkyl, -CN , F, C _1-4 -alkyl;

where in (iv) it is preferable

R ³ represents a group selected from H, -PO3R ^3a 2, -CH ₂ -OPO ₃ R ^3a ₂ and -CH ₂ -OC(=O)-R ^3b ;

R ^3a represents a hydrogen atom or cation suitable to form a pharmaceutically acceptable salt of either -CH2-OC(=O)-R ^3b or -CHMe-OC(=O)-R ^3b or -CMe2-OC(=O)-R ^3b or -CH ₂ -OC(=O)-OR ^3b or -CHMe-OC(=O)-OR ^3b or -CMe2-OC(=O)-OR ^3b ;

R ^3b represents an alkyl group having from 1 to 11 carbon atoms, which is linear, branched or cyclic, or a combination thereof, wherein the R ^3b group is optionally substituted by one or more groups independently selected from -OH and -OC _1-6 -alkyl ;

R ⁴ represents H, Ci-4-alkyl; And

R ⁵ represents H, Ci-4-alkyl, C _1-4 -alkylene-OR ³ ;

R ⁶ represents H, Ci-4-alkyl, OH, -OPO3R ^3a 2; And

R ⁷ is O^-alkyl, C1-4-alkylene-OH, C1-4-alkylene-OR ³ , where R ³ is as defined above, C1-4-alkylene-F;

R ⁸ represents a group selected from -O-Ar ² and -O-Het ² , where Ar ² represents a phenyl group that is optionally substituted by one or more groups selected from -CN, -OC _1-4 -alkyl, or where phenyl group may bear two substituent groups on adjacent ring atoms such that these adjacent substituent groups can be linked together to form a 5-membered heterocycle having one or two heteroatoms selected from N, S and O, wherein said heterocycle may optionally bear one or two substituents individually selected from oxo, F, -OC _1-4 -alkyl, Ci-4-alkyl. CN; and where Het ² represents an aromatic heterocycle with 5 or 6 ring atoms, including 1 or 2 heteroatoms individually selected from N, S and O, which may optionally be substituted by one or more groups individually selected from -OC _1-4 -alkyl, -CN, F, C _1-4 -alkyl.

Embodiment 15. A pharmaceutical composition comprising the compound of any of embodiments 1-14 and a pharmaceutically acceptable carrier.

Embodiment 16. Use of the compound of any one of embodiments 1 to 14 or the pharmaceutical composition of claim 15 for the treatment of a bacterial infection selected from infections caused by N. gonorrhoeae, Chlamydia trachomatis, or a combination thereof.

Definitions.

The following definitions are intended to assist the reader. Unless otherwise indicated, all technical terms, symbols and other scientific or medical terms or terminology used herein have the meanings commonly understood by those skilled in the art of chemistry and medicine. In some cases, terms with generally accepted meanings are defined herein for clarity and/or ease of reference, and the inclusion of such definitions herein should not be construed as representing a material difference from the definition of the term as generally understood in the art.

In some embodiments, the term approximately refers to a deviation of ±10% from the stated value. When the word about is used herein in relation to a number, it should be understood that another embodiment of the invention includes that number unmodified by the presence of the word about.

Administering or administering a drug to a patient (and the grammatical equivalents of this phrase) refers to direct administration, which can be the administration of a drug to a patient by a health care professional, or can be self-administration and/or indirect administration, which can be the act of prescribing a drug. For example, a physician who instructs a patient to self-administer a drug or gives a patient a prescription for a drug administers the drug to the patient.

Dose and dosage refer to the specific amount of active or therapeutic agents to be administered. Such amounts are included in a dosage form, which refers to physically discrete units suitable as unitary dosages for humans and other mammals, wherein each unit contains a predetermined amount of active agent calculated to produce the desired onset, tolerability and therapeutic effects, in combination with one or several suitable pharmaceutical excipients, such as carriers.

The terms treatment and therapy as used herein refer to a variety of hygienic, pharmacological, surgical and/or physical means used with the intent to cure and/or alleviate disease and/or symptoms in order to solve a health problem. The terms treatment and therapy include preventive and curative methods, as both are aimed at maintaining and/or restoring the health of an individual or animal. Regardless of the origin of the symptoms, disease or disability, administering the appropriate medication to

- 10 046887 relief and/or treatment of a health problem should be interpreted as a form of treatment or therapy in the context of this application.

Unit dosage form as used herein refers to a physically discrete unit of therapeutic composition suitable for the subject being treated. However, it should be understood that the total daily use of the compositions of the present invention will be determined by the attending physician within the bounds of sound medical judgment. The specific effective dose level for any particular subject or organism will depend on a variety of factors, including the disease being treated and the severity of the disorder; the activity of the particular active agent used; the specific composition used; age, body weight, general health, gender, and diet of the subject; timing of administration and rate of elimination of the particular active agent used; duration of treatment; drugs and/or additional therapies used in combination or concurrently with the particular compound(s) used, and similar factors well known in the medical field.

For example, element means one element or more than one element.

The term including is used to mean including, but not limited to. Including and including but not limited to are used interchangeably. The term containing is used in the same sense as including. The term consisting of is used to indicate that the listed elements are present, but other, not mentioned elements are absent. The term comprising is used to include the meaning consisting of, as a preferred embodiment.

The term FabI is known in the art and refers to a bacterial enzyme that is believed to function as an enoyl acyl carrier protein (ACP) reductase in the final step of the four reactions involved in each cycle of bacterial fatty acid biosynthesis. This enzyme is believed to be widespread in bacteria and plants.

The term enzyme inhibitor refers to any compound that prevents an enzyme from effectively performing its respective biochemical roles. Therefore, a FabI inhibitor is any compound that inhibits FabI from performing its biochemical role. The degree of enzyme inhibition by any such compound will vary and is described herein and elsewhere.

The term antibiotic agent or antibacterial agent shall mean any drug that is useful for treating, preventing, or otherwise reducing the severity of any bacterial disease or any of its complications, including any of the conditions, diseases, or complications resulting from and/or described herein. Antibiotic agents include, for example, cephalosporins, quinolones and fluoroquinolones, penicillins and beta-lactamase inhibitors, carbapenems, monobactams, macrolides and lincosamides, glycopeptides, rifampin, oxazolidinones, tetracyclines, aminoglycosides, streptogramins, sulfonamides and the like. Other antibiotic or antibacterial agents are described herein and are known to those skilled in the art. In some embodiments, the term antibiotic agent does not include an agent that is a FabI inhibitor, so the combinations of the present invention will in some cases include one agent that is a FabI inhibitor and another agent that is not a FabI inhibitor.

The term medicinal product as used in this document refers to any substance falling within at least one of the definitions given in Article 1(b), 2(b) or 3a of Directive 2001/83/EC of 6 November 2001, as amended on November 16, 2012, or in Article 1, paragraph 2(h) or 2(b) of Directive 2001/82/EC of November 6, 2001, as amended on August 7, 2009 and in Article 2 of Regulation (EC) No. 726/2004 of March 31, 2004

The term disease as used herein refers to any disease caused by or associated with infection of the body.

The term bacterial disease as used herein refers to any disease caused by or associated with infection by bacteria.

The term cis is a common one in the art and refers to the arrangement of two atoms or groups around a double bond such that the atoms or groups are on the same side of the double bond. Cis configurations are often referred to as (Z) configurations.

The term trans is a common one in the art and refers to the arrangement of two atoms or groups around a double bond such that the atoms or groups are on opposite sides of the double bond. trans Configurations are often referred to as (E) configurations.

The term therapeutic effect is accepted in the art, and refers to the local or systemic effect in animals, in particular mammals, and more particularly in humans, caused by a pharmacologically active substance. Thus, the term means any measurable effect in the diagnosis, treatment, mitigation, treatment or prevention of disease or in the improvement of desired physical or mental development and/or conditions in an animal or human. The phrase therapeutically effective amount means that amount of such a substance that produces

- 11 046887 desired local or systemic effect with a reasonable benefit/risk ratio applicable to any treatment. A therapeutically effective amount of such a substance will vary depending on the subject and the disease state being treated, the body weight and age of the subject, the severity of the disease condition, the route of administration, and the like, as can be readily determined by one skilled in the art. For example, certain compositions of the present invention may be administered in an amount sufficient to produce a therapeutic effect at a reasonable benefit/risk ratio applicable to such treatment.

The term chiral is common in the art and refers to molecules that have the property of not matching when mirrored, while the term achiral refers to molecules that are matching when mirrored. A prochiral molecule is a molecule that has the potential to be converted into a chiral molecule in a specific process.

The compounds of the present disclosure may contain one or more chiral centers and/or double bonds and, therefore, exist as geometric isomers, enantiomers or diastereomers. Enantiomer and diastereomers may be designated by the symbols (+), (-), R or S depending on the configuration of substituents around the stereogenic carbon atom, but one skilled in the art will appreciate that the structure may implicitly denote one or more chiral centers. Mixtures of enantiomers or diastereomers may be designated by the nomenclature (±), but one skilled in the art will recognize that the structure may implicitly denote a chiral center. Geometric isomers resulting from the arrangement of substituents around a carbon-carbon double bond or the arrangement of substituents around a cycloalkyl or heterocyclic ring may also exist in the compounds of the present invention.

The symbol ............ denotes a bond, which may be a single, double, or triple bond as described herein.

Substituents around the carbon-carbon double bond are designated as having the Z or E configuration, where the terms Z and E are used in accordance with IUPAC standards. Unless otherwise noted, structures depicting double bonds cover both the E and Z isomers. Substituents around a carbon-carbon double bond may alternatively be referred to as cis or trans, where cis represents substituents on the same side of the double bond and trans represents substituents on opposite sides of the double bond. The arrangement of substituents around the carbocyclic ring can also be designated as cis or trans. The term cis represents substituents on the same side of the ring plane, and the term trans represents substituents on opposite sides of the ring plane. Mixtures of compounds in which substituents are located on both the same and opposite sides of the ring plane are designated cis/trans or Z/E.

The term stereoisomers as used herein consists of all geometric isomers, enantiomers or diastereomers. The present invention covers various stereoisomers of these compounds and mixtures thereof. Conformational isomers and rotamers of the described compounds are also considered.

The term ED50 is accepted in the art. In some embodiments, ED50 means the effective dose of a drug that produces 50% of its maximal response or effect, or alternatively, the dose that produces a predetermined response in 50% of test subjects or drugs. The term LD50 is accepted in the art. In some embodiments, the LD50 refers to the dose of drug that is lethal in 50% of subjects. The term therapeutic index is an accepted term in the art that refers to the therapeutic index of a drug, defined as ED50/LD50.

The term Ki is common in the art and refers to the dissociation constant of the enzyme-inhibitor complex.

The term antimicrobial is a common one in the art and refers to the ability of the compounds described herein to prevent, inhibit or destroy the growth of microbes such as bacteria, fungi, protozoa and viruses.

The term antibacterial is a common one in the art and refers to the ability of the compounds disclosed herein to prevent, inhibit or kill the growth of microbial bacteria.

The term microbe is a common one in the art and refers to a microscopic organism. In some embodiments, the term microbe is applied to bacteria. In other embodiments, the term refers to pathogenic forms of a microscopic organism.

The term alkyl, as used herein, refers to a saturated straight or branched hydrocarbon, such as a straight or branched group of 1-8 or 1-6 carbon atoms, designated herein as C1- _C8 alkyl or C-^-alkyl, respectively. The term lower alkyl as used herein more specifically refers to a saturated straight or branched hydrocarbon, such as a straight or branched group of 1-4 or 1-3

- 12,046,887 carbon atoms, designated herein as C1- _C4 alkyl and C1-C3 alkyl, respectively. Typical alkyl groups and lower alkyl groups include, but are not limited to, methyl, ethyl, propyl, isopropyl, 2-methyl-1-propyl, 2-methyl-2-propyl, 2-methyl-1-butyl, 3-methyl-1 -butyl, 3-methyl-2-butyl, 2,2-dimethyl-1-propyl, 2-methyl-1-pentyl, 3-methyl-1-pentyl, 4-methyl-1-pentyl, 2-methyl-2 -pentyl, 3methyl-2-pentyl, 4-methyl-2-pentyl, 2,2-dimethyl-1-butyl, 3,3-dimethyl-1-butyl, 2-ethyl-1-butyl, butyl, isobutyl, t -butyl, pentyl, isopentyl, neopentyl and hexyl.

In addition, the term alkyl (or lower alkyl) includes substituted alkyls, i.e. it should be understood as optionally bearing one or more substituents in one or more positions. That is, it also refers to alkyl groups having one or more (eg, two, three, four, five, six, etc.) substituents, each of which replaces a hydrogen on a carbon atom of the hydrocarbon backbone. Such substituents may include, for example, a hydroxyl group, a carbonyl group (where the carbonyl group carries a hydrogen atom, an alkyl group or other group as defined in this paragraph, for example, to give a carboxyl, alkoxycarbonyl, formyl or acyl group), a thiocarbonyl group (where the carbonyl group carries a hydrogen atom, an alkyl group or other group as defined in this paragraph, for example, to give a thioether, thioacetate or thioformate), alkoxy, phosphoryl, phosphonate, phosphinate, phosphate, amino, amido, amidine, imine, cyano, nitro, azido, sulfhydryl, alkylthio, sulfate, sulfonate, sulfamoyl, sulfonamido, sulfonyl, heterocyclyl, aralkyl, cycloalkyl, heterocycle or an aromatic or heteroaromatic group. In all cases where the above groups have more than one valency, the additional free valency may be saturated with a hydrogen atom, an alkyl group, a cycloalkyl group, a heterocyclic group, an aryl group, or a heteroaryl group. In addition, those skilled in the art will appreciate that moieties substituted on the hydrocarbon chain may themselves be substituted, if applicable. For example, substituted alkyl substituents may include substituted and unsubstituted forms of amino, azido, imino, amido, phosphoryl (including phosphonate, phosphinate and phosphate), sulfonyl (including sulfate, sulfonamido, sulfamoyl and sulfonate) and silyl groups, as well as ethers, alkylthio , carbonyls (including ketones, aldehydes, carboxylates and esters), nitrile and isonitrile. For the avoidance of doubt, an alkyl group carrying another alkyl group should not be considered as an alkyl group substituted by another alkyl group, but as a single branched alkyl group.

The term alkenyl is a common one in the art and refers to a group corresponding to an alkyl group as defined above but bearing one or more carbon-carbon double bonds. Of course, the total number of double bonds is limited by the number of carbon atoms in the alkenyl group, and in order to account for at least one double bond, the alkenyl group must have at least two carbon atoms. Apart from this difference, the definitions and characteristics given for the alkyl group above apply equally to the alkenyl group.

The term alkynyl is a common one in the art and refers to a group corresponding to an alkyl group as defined above but bearing one or more carbon-carbon triple bonds. Of course, the total number of double bonds is limited by the number of carbon atoms in the alkenyl group and in order to account for at least one triple bond, the alkynyl group must have at least two carbon atoms. Apart from this difference, the definitions and characteristics given for the alkyl group above apply equally to the alkynyl group.

The term aryl is a common one in the art and refers to 5-, 6-, and 7-membered single-ring aromatic groups that may contain zero to four heteroatoms, e.g., benzene, pyrrole, furan, thiophene, imidazole, oxazole, thiazole, triazole , pyrazole, pyridine, pyrazine, pyridazine and pyrimidine and the like. Those aryl groups that have heteroatoms in the ring structure may also be called heteroaryl or heteroaromatic compounds. The aromatic ring may be substituted at one or more positions on the ring with such substituents as described above, for example, halogen, azide, alkyl, aralkyl, alkenyl, alkynyl, cycloalkyl, hydroxyl, alkoxyl, amino, nitro, sulfhydryl, imino, amido, phosphonate, phosphinate, phosphate, carbonyl, carboxyl, silyl, ether, alkylthio, sulfonyl, sulfonamido, ketone, aldehyde, ester, heterocyclyl, aromatic or heteroaromatic groups, -CF ₃ , -CN and the like. The term aryl also includes polycyclic ring systems containing two or more cyclic rings in which two or more carbon atoms are common to two adjacent rings (the rings are fused rings), where at least one of the rings is aromatic, e.g. the cyclic rings may be cycloalkyls, cycloalkenyls, cycloalkynyls, aryls and/or heterocyclyls.

The term aralkyl or arylalkyl is common in the art and refers to an alkyl group substituted with an aryl group (eg, an aromatic or heteroaromatic group).

- 13 046887

The term carbocycle is a common one in the art and refers to an aromatic or non-aromatic ring in which each ring atom is a carbon.

The term cycloalkyl as used herein refers to a monocyclic saturated or partially unsaturated alkyl or alkenyl group of, for example, 3-6 or 4-6 carbon atoms, referred to herein as, for example, '^-cycloalkyl or _{C4- 6} cycloalkyl, and is derived from a cycloalkane. Examples of cycloalkyl groups include, but are not limited to, cyclohexane, cyclohexene, cyclopentane, cyclobutane, cyclopropane or cyclopentene. Said cycloalkyl group may be substituted at one or more positions with one or more substituents as described above.

The terms halo or halogen as used herein refer to F, Cl, Br or I. Halide refers to the corresponding anion of the halogens, and pseudohalide is defined on page 560 of Advanced Inorganic Chemistry by Cotton and Wilkinson, Interscience Publishers, 1966.

The term amino as used herein refers to any group of the structure NR _a R _b where, unless otherwise indicated, Ra and R _b are independently selected from the group consisting of H, alkyl, cycloalkyl, alkenyl, alkynyl, aryl, heteroaryl, heterocyclic groups, as well as any other groups of substituents listed above with respect to the scope of substituted alkyl groups, with the exception of carbonyl groups, thiocarbonyl groups, imine groups and substituent groups in which the attachment to the remaining molecule is carried out through a heteroatom selected from N, O, S and P. Alternatively, Ra and Rb may be hydrocarbon groups that are bonded to form a heterocycle together with the nitrogen atom to which they are attached.

The term heteroaryl as used herein refers to a monocyclic aromatic 4 to 6 membered ring system containing one or more heteroatoms, such as one to three heteroatoms, which may be the same or different, such as nitrogen, oxygen and sulfur. If possible, said heteroaryl ring may be linked to an adjacent radical via carbon or nitrogen. Examples of heteroaryl rings include, but are not limited to, furan, benzofuran, thiophene, pyrrole, thiazole, oxazole, isothiazole, isoxazole, imidazole, pyrazole, triazole, pyridine and pyrimidine. Said heteroaryl group may be substituted with one or more substituents as described for the aryl group above. The term heteroaryl also includes polycyclic ring systems having two or more cyclic rings in which two or more carbon atoms or heteroatoms are common to two adjacent rings (the rings are fused rings) wherein at least one of the rings is heteroaryl, as defined above, while other cyclic rings may be cycloalkyls, cycloalkenyls, cycloalkyls, aromatic rings and/or saturated, unsaturated or aromatic heterocycles.

The term heterocycle as used herein refers to a monocyclic ring containing one or more heteroatoms, such as one to three heteroatoms, which may be the same or different, such as nitrogen, oxygen and sulfur. The remaining members of the ring are formed by carbon atoms. The heterocycle typically has 4 to 8 ring members and preferably 5 or 6 ring members. Unless otherwise specified, the heterocycle may be aromatic, partially or fully saturated. Unless otherwise indicated, it may or may not contain valid substituents as specified herein.

The terms hydroxy and hydroxyl as used herein refer to the OH radical.

The term nitro is a common one in the art and refers to -NO2; the term sulfhydryl is common in the art and refers to -SH; and the term sulfonyl is as accepted in the art and refers to -SO2-.

The definition of each expression, when it occurs more than once in any structure, must be independent of its definition anywhere else in the same structure.

The terms triflyl, tosyl, mesyl and nonaphlyl are common in the art and refer to trifluoromethanesulfonyl, p-toluenesulfonyl, methanesulfonyl and nonafluorobutanesulfonyl groups, respectively. The terms triflate, tosylate, mesylate and nonaflate are common in the art and refer to the trifluoromethanesulfonate, p-toluenesulfonate, methanesulfonate and nonafluorobutanesulfonate functional groups and molecules that contain these groups, respectively.

The abbreviations Me, Et, Ph, Tf, Nf, Ts and Ms represent methyl, ethyl, phenyl, trifluoromethanesulfonyl, nonafluorobutanesulfonyl, p-toluenesulfonyl and methanesulfonyl, respectively. A more complete list of abbreviations used by organic chemists is provided in the first issue of each volume of the Journal of Organic Chemistry; this list is usually presented in a table called Standard List of Abbreviations.

The term prodrug refers to a derivative of an active compound (drug) that undergoes transformation under conditions of use, such as in the body, to release

- 14 046887 nium of the active drug. Prodrugs are often, but not necessarily, pharmacologically inactive until they are converted to an active drug.

It should be understood that substitution or substituted includes the implicit condition that such substitution corresponds to the allowed valency of the substituted atom and substituent, and that the substitution results in a stable compound, for example, in which spontaneous transformation such as rearrangement, cyclization, elimination or other reactions does not occur.

The term substituted is also considered to include all valid substituents for organic compounds. In broad terms, acceptable substituents include acyclic and cyclic, branched and straight-chain, carbocyclic and heterocyclic, aromatic and non-aromatic substituents of organic compounds. Exemplary substituents include, for example, those described herein above, for example, in connection with substituted alkyls. Acceptable substituents may be one or more and may be the same or different for suitable organic compounds. For purposes of this description, heteroatoms, such as nitrogen, may have hydrogen substituents and/or any permissible substituents of the organic compounds described herein that satisfy the valency of the heteroatoms. In this context, the term acceptable substituents means any substituent that can be bonded to the parent molecule without violating the general principles of chemical bonding, such as the maximum number of valence electrons for the atom of interest, and without making the compound so toxic to the patient that unacceptable toxicity is found even at the minimum dosage required to achieve a therapeutic effect.

For the purposes of this invention, the chemical elements are identified in accordance with the Periodic Table of the Elements, CAS version, Handbook of Chemistry and Physics, 67th Ed., 1986-87, inside the cover. Also for purposes of description, the term hydrocarbon is considered to include all valid compounds having at least one hydrogen atom and one carbon atom. In broad terms, acceptable hydrocarbons include acyclic and cyclic, branched and unbranched, carbocyclic and heterocyclic, aromatic and non-aromatic organic compounds, which may be substituted or unsubstituted.

The term pharmaceutically acceptable salts is a common one in the art and refers to relatively non-toxic inorganic and organic acid addition salts or inorganic or organic base addition salts, including, for example, those contained in the compositions of the present invention, and including those present in others. approved medicinal products (where approval may be given by any competent authority in the EU, USA, CA, JP, CN or KR). Pharmaceutically acceptable salts are intended to be included within the scope of the present invention. Therefore, all references to the compounds of the present invention should be understood as references not only to the compounds themselves, but also to pharmaceutically acceptable salts of the corresponding compounds. In one aspect, the pharmaceutically acceptable salts may be selected from those recognized as pharmaceutically acceptable in the literature on the date of filing, and in particular as described in G.S. Paulekuhn et al. in J. Med. Chem. 2007, 50, 6665-6672 and references cited therein. If the compound of the invention is an acidic compound, and in particular a prodrug containing a phosphate group, the compound of the invention may be provided in the form of pharmaceutically acceptable salts, wherein the compound forms an anionic moiety and the counterion is selected from Na, K, Mg, Ca, or protonated forms of the organic bases ethanolamine, meglumine, tromethamine (i.e. 2-amino-2-(hydroxymethyl)propane-1,3-diol) and deanol (i.e. 2-(dimethylamino)ethanol). The stoichiometry of the salts is not particularly limited. For example, salts can be formed with the phosphate prodrugs of the invention (having 2 acidic protons) in any stoichiometric ratio from 0 to 2 equivalents. Typically, pharmaceutically acceptable salts are formed such that the net charge on the salt is zero, i.e. that the total number of positive charges is equal to the total number of negative charges.

The term treatment includes any significant effect, such as reduction, reduction, modulation or elimination, which results in the improvement of a condition, disease, disorder and the like.

The term prophylactic or therapeutic treatment is accepted in the art and refers to the administration of one or more subject compositions to the host. If administration occurs before the clinical manifestation of an unwanted condition (e.g., a disease or other adverse condition in the host animal), then the treatment is prophylactic, that is, it protects the host from developing the unwanted condition, while if administration occurs after the manifestation of the unwanted condition, the treatment is therapeutic (i.e. it is intended to reduce, alleviate or maintain an existing undesirable condition or associated side effects).

The patient, subject or host being treated by this method may mean either a human or a non-human animal. Non-human animals include companion animals (eg cats, dogs) and animals raised for food (ie food animals) such as cows, pigs, chickens. Non-human animals are preferred

- 15 046887 are definitely mammals.

The term mammal is known in the art, and exemplary mammals include humans, primates, cattle, pigs, dogs, cats, and rodents (eg, mice and rats).

The term bioavailable is accepted in the art and refers to a form of the subject matter that allows it, or a portion of the administered amount, to be absorbed, administered, or otherwise be physiologically available to the subject or patient to whom it is administered.

The term pharmaceutically acceptable carrier is accepted in the art and refers to a pharmaceutically acceptable material, composition or carrier, such as a liquid or solid excipient, diluent, excipient, solvent or encapsulating material involved in moving or transporting any subject composition or component thereof from one organ or body part into another organ or body part. Each carrier must be acceptable in the sense that it is compatible with the subject composition and its components and does not cause harm to the patient. Some examples of materials that may serve as pharmaceutically acceptable carriers include: (1) sugars such as dextrose, lactose, glucose and sucrose; (2) starches such as corn starch and potato starch, as well as starch derivatives such as cyclodextrins and modified cyclodextrins, including preferably (2-hydroxypropyl)-β-cyclodextrin and sulfobutyl ether-β-cyclodextrin; (3) cellulose and its derivatives, such as microcrystalline cellulose, sodium carboxymethylcellulose, methylcellulose, ethylcellulose, hydroxypropylmethylcellulose (HPMC) and cellulose acetate; (4) powdered tragacanth; (5) malt; (6) gelatin; (7) talc; (8) matrix-forming polymer excipients such as polyvinylpyrrolidine (PVP), for example PVP K30, acrylic polymers and copolymers, such as various grades of Eudragit and, preferably, Eurdragit L100, hydroxypropyl methylcellulose succinate (HPMCAS), other copolymers, such as copolymers based on polyethylene glycol, such as Soluplus; (9) excipients such as cocoa butter and suppository waxes; (10) oils such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil and soybean oil; (11) glycols such as propylene glycol; (12) polyols such as glycerin, sorbitol, mannitol and polyethylene glycol; (13) esters such as ethyl oleate, glyceryl behenate and ethyl laurate; (14) agar; (15) buffering agents such as magnesium hydroxide and aluminum hydroxide; (16) alginic acid; (17) pyrogen-free water; (18) isotonic saline solution; (19) Ringer's solution; (20) ethyl alcohol; (21) phosphate buffer solutions; and (22) other non-toxic compatible substances used in pharmaceutical formulations. The excipients described may perform more than one function. For example, fillers or binding agents may also be disintegrating agents, glidants, antiblocking agents, lubricants, sweeteners, and the like.

The term solvent is used herein to mean a liquid chemical that is capable of dissolving a significant amount of another substance of interest, a solute, to form a clear, homogeneous solution. The term significant amount is defined by the intended use of the solution such that the intended use must be possible due to the dissolved amount of solute. For example, if a compound of the present invention is to be administered in the form of a solution by injection, the solvent must be capable of dissolving the compound in such quantities as to permit administration of a therapeutic dose.

Unless otherwise indicated, all reactions described herein are carried out at reaction temperatures that yield the desired target compound and that provide a reasonable compromise between reaction rate and selectivity. Typical reaction temperatures for Pd-based coupling reactions and Fe-based cyclization reactions range from 80°C to 90°C, while deprotection is typically carried out at temperatures from 0°C to room temperature (25°C).

Unless otherwise indicated, all characteristics of the variable groups are the same as those indicated for the compound of formula I, or, likewise, they are to be understood so that more specific meanings and combinations of meanings represented by formulas II-XVI are also possible and even preferred.

Unless otherwise specified, the term protecting group is used herein to characterize a group that is bonded to a functional group to prevent that functional group from participating in the intended chemical reaction. The protecting group must be inert under the conditions of the intended chemical reaction, but it must be possible to remove the protecting group from the compound without further transformations occurring in other parts of the molecule. Suitable protecting groups are described for each functional group in Greene's Protective Groups in Organic Synthesis, Peter G. M. Wuts, Theodora W. Greene, John Wiley & Sons, 20 Dec 2012.

Compounds of the invention.

The present invention provides compounds that exhibit beneficial antibiotic activity through FabI inhibition. The compounds of the present invention are represented by the following general formula I

- 16 046887

ABOUT

I 3

R I where A1 represents a group selected from groups A11 and A12 having the following structures:

where the line connected to the exocyclic methylene group represents a single covalent bond formed with the nitrogen atom of formula I;

A2 represents a methyl group;

or A1 and A2 together with the nitrogen atom to which they are bonded form the following group A3:

or it may be any mixture thereof, including a particularly racemic mixture of the above two enantiomers.

Any of these groups may be designated as group A3;

and where in option (iii)

Q1 represents CH2 or NH;

Q2 represents CR ⁴ R ⁵ -CR ⁶ R ⁷ where the CR4R ⁵ group binds to Q1;

or Q1-Q2 is -N=CR ⁷ -CHR ⁷ - or its tautomer -NH-CR ⁷ =CR ⁷ -;

Q3 represents O or S;

R ¹ represents a group selected from H, -NH ₂ ;

R2 represents a group selected from H, -O-Ar ¹ , -O-Het ¹ , -NR ⁹ R ¹⁰ , -O-Alk ¹ where Ar ¹ represents a phenyl group which may be optionally substituted by one or more groups selected of -CN, -OC _1-4 -αalkyl, -O-(CH2) _1-4 -NR ⁹ R ¹⁰ , or where the phenyl group may bear two substituent groups on adjacent ring atoms so that these adjacent substituent groups can be bonded

- 17 046887 are combined to form a 5-membered heterocycle having one or two heteroatoms individually selected from N and O, where Het ¹ is an aromatic heterocycle with 5 or 6 ring atoms containing 1 or 2 heteroatoms individually selected from N, S and O, or a non-aromatic partially or fully saturated heterocycle with 6 ring atoms including 1 heteroatom selected from N and O, wherein the Het ¹ group is optionally substituted by one or more groups individually selected from -C.'i-.-1- alkyl -O-C1-4-alkyl, -CN, -(CH2)0-4-OH; where R ⁹ is selected from H and -C1-4alkyl; where R ¹⁰ is selected from H, -C _1-4 -alkyl and -C(=O)-CH3; where Alk ¹ represents an alkyl group having from 1 to 6 carbon atoms, which is linear, branched or cyclic, or a combination thereof, where the Alk ¹ group is optionally substituted by one or more groups selected from -OH, -O-alkyl;

R3 represents a group selected from H, -PO3R ^3a 2, -CH2-OPO3R ^3a 2 and -CH ₂ -OC(=O)-R ^3b ;

R ^3a represents a hydrogen atom or cation suitable to form a pharmaceutically acceptable salt of either -CH2-OC(=O)-R ^3b or -CHMe-OC(=O)-R ^3b or -CMe2-OC(=O)-R ^3b or -CH ₂ -OC(=O)-OR ^3b or -CHMe-OC(=O)-OR ^3b or -CMe2-OC(=O)-OR ^3b ;

R ^3b represents an alkyl group having from 1 to 11 carbon atoms, which is linear, branched or cyclic, or a combination thereof, wherein the R ^3b group may optionally be substituted by one or more groups independently selected from -OH and -O^^- alkyl;

R ⁴ represents a group selected from H, C _1-4 -alkyl, CN and C _1-4 -alkylene-F;

R ⁵ represents a group selected from H, C _1-4 -alkyl, CY.-i-alkylene-OH, C _1-4 -alkylene-OR ³ , -OH, -OPO3R ^3a 2;

or R ⁴ and R ⁵ together form a cyclic group having from 4 to 6 ring members formed by methylene groups and, optionally, an oxygen atom;

R ⁶ represents a group selected from -OH, C1-4-alkyl, -OPO3R ^3a 2;

R ⁷ represents a group selected from H, C1-4-alkyl, C1-4-alkylene-OH, C1-4-alkylene-OR ³ , C1-4-alkylene-F, -CN;

or R ⁶ and R ⁷ together form a cyclic group having from 4 to 6 ring members formed by methylene groups and, optionally, an oxygen atom; wherein the cyclic group may optionally bear a substituent selected from -OH, -O-alkyl;

R ⁸ represents a group selected from -O-Ar ² and -O-Het ² ;

where Ar ² represents a phenyl group which is optionally substituted by one or more groups selected from -CN, -OC _1-4 -alkyl, or where the phenyl group may bear two substituent groups on adjacent ring atoms such that these adjacent substituent groups may be bonded together to form a 5-membered heterocycle having one or two heteroatoms individually selected from N, S and O, wherein said heterocycle may optionally bear one or two substituents selected from oxo, halogen, -O-^l-alkyl, Cj -.galkyla and CN; and where Het ² represents an aromatic heterocycle with 5 or 6 ring atoms, including 1 or 2 heteroatoms individually selected from N, S and O, which may optionally be substituted by one or more groups individually selected from -OC _1-4 -alkyl, -CN, F, C _1-4 -alkyl, and pharmaceutically acceptable salts thereof; or in option (iv)

Q1 represents CH2 or NH;

Q2 represents CR ⁴ R ⁵ -CR ⁶ R ⁷ where the CR4R ⁵ group binds to Q1;

or Q1-Q2 is -N=CR ⁷ -CHR ⁷ - or its tautomer -NH-CR ⁷ =CR ⁷ -;

Q3 represents O or S;

R ¹ represents a group selected from H, -NH2;

R ² represents a group selected from H, -O-Ar ¹ , -O-Het ¹ , -NR ⁹ R ¹⁰ , -O-Alk ¹ where Ar ¹ represents a phenyl group which is optionally substituted by one or more groups selected from -CN, -O-C1-4alkyl, -O-(CH2)1-4-NR ⁹ R ¹⁰ , or where the phenyl group may bear two substituent groups on adjacent ring atoms such that these adjacent substituent groups may be bonded together forming a 5-membered heterocycle having one or two heteroatoms individually selected from N and O, where Het ¹ is an aromatic heterocycle with 5 or 6 ring atoms containing 1 or 2 heteroatoms individually selected from N, S and O, or non-aromatic a partially or fully saturated heterocycle with 6 ring atoms containing 1 heteroatom selected from N and O, where the Het ¹ group may optionally be substituted by one or more groups individually selected from ^l-alkyl, -O^l-alkyl, -CN , -(CH2) _0-4 -OH; where R ⁹ is selected from H and ^l-alkyl; where R ¹⁰ is selected from H, ^l-alkyl and -C(=O)-CH ₃ ; where Alk ¹ represents an alkyl group having from 1 to 6 carbon atoms, which is linear, branched or cyclic, or a combination thereof, where the Alk ¹ group may be optionally substituted by one or more groups selected from -OH, -O-alkyl;

R ³ represents a group selected from H, -PO3R ^3a 2, -CH ₂ -OPO ₃ R ^3a ₂ and -CH ₂ -OC(=O)-R ^3b ;

R ^3a represents a hydrogen atom or cation suitable to form a pharmaceutically acceptable salt of either -CH2-OC(=O)-R ^3b or -CHMe-OC(=O)-R ^3b or -CMe2-OC(=O)-R ^3b or -CH ₂ -OC(=O)-OR ^3b or -CHMe-OC(=O)-OR ^3b or -CMe2-OC(=O)-OR ^3b ;

- 18 046887

R ^3b represents an alkyl group having from 1 to 11 carbon atoms, which is linear, branched or cyclic, or a combination thereof, wherein the R ^3b group is optionally substituted by one or more groups independently selected from -OH and -OC _1-6 -alkyl ;

R ⁴ represents a group selected from H, C _1-4 -alkyl, CN and C _1-4 -alkylene-F;

R ⁵ represents a group selected from H, C _1-4 -alkyl, C _1-4 -alkylene-OH, C _1-4 -alkylene-OR ³ , -OH, -OPO.R.'· ¹ ,;

or R ⁴ and R ⁵ together form a cyclic group having from 4 to 6 ring members formed by methylene groups and, optionally, an oxygen atom;

R ⁶ represents a group selected from H, -OH, C1-4-alkyl, -OPO3R ^3a 2;

R ⁷ represents a group selected from C1-4-alkyl, C1-4-alkylene-OH, C1-4-alkylene-OK ³ , C1-4-alkylene-G, -CN;

or R ⁶ and R ⁷ together form a cyclic group having from 4 to 6 ring members formed by methylene groups and, optionally, an oxygen atom; wherein the cyclic group may optionally bear a substituent selected from -OH, -O-alkyl;

R ⁸ represents a group selected from -Ο-Ar ² and -O-Het ² ;

where Ar ² represents a phenyl group which is optionally substituted by one or more groups selected from -CN, -O-C _1-4 -alkyl, or where the phenyl group may bear two substituent groups on adjacent ring atoms such that these adjacent substituent groups may be linked together to form a 5-membered heterocycle having one or two heteroatoms individually selected from N, S and O, wherein said heterocycle may optionally bear one or two substituents selected from oxo, halogen, -O-C _1-4 -alkyl, C _1-4 -alkyl and CN; and where Het ² represents an aromatic heterocycle with 5 or 6 ring atoms, comprising 1 or 2 heteroatoms individually selected from N, S and O, which may optionally be substituted by one or more groups individually selected from -O-C _1-4 - alkyl, -CN, F, C _1-4 -alkyl, and pharmaceutically acceptable salts thereof,

In one embodiment, the compounds of the present invention are represented by the general formula II

where R ¹ , R ² , R ³ , Q1, Q ₂ , Q ₃ have the same meanings as defined for the general formula I above;

and wherein the five-membered heterocycle containing _Q3 is bonded to a methyleneamide group at position 2 and a methyl group at position 3, or is bonded to a methyleneamide group at position 3 and a methyl group at position 2.

Preferably, the compounds of this embodiment are represented by the general formula III

where R ¹ , R ² , R ³ , Q1, Q ₂ , Q ₃ have the same meanings as defined for the general formula I above.

According to another aspect of this embodiment, the group variables Q1 and Q ₂ represent NH and CR ⁴ R ⁵ -CR ⁶ R ⁷ , respectively. Compounds in this aspect are represented by the general formula VI

R VI where R ¹ , R ² , R ³ , R ⁴ , R ⁵ , R ⁶ and R ⁷ have the same meanings as defined for general formula I above in option (iii) or (iv) above, where in option (iii) preferably

R ¹ represents H, NH ₂ ;

R ² represents H;

R ³ represents a group selected from H, -PO ₃ R ^3a 2, -CH2-OPO3R ^3a 2 and -CH ₂ -OC(=O)-R ^3b ;

R ^3a represents a hydrogen atom or a cation suitable for forming a pharmaceutically acceptable salt or -CH2-OC(=O)-R ^3b or -CHMe-OC(=O)-R ^3b or -CMe2-OC(=O )-R ^3b or -CH2-OC(=O)-OR ^3b or -CHMe-OC(=O)-OR ^3b or -CMe ₂ -OC(=O)-OR ^3b ;

R ^3b represents an alkyl group having from 1 to 11 carbon atoms, which is linear, branched or cyclic, or a combination thereof, wherein the R ^3b group is optionally substituted by one or more groups independently selected from -OH and -O-Cm-alkyl;

R ⁴ represents H, Cm-alkyl; And

R ⁵ represents H, C _1-4 -alkyl, C _1-4 -αalkylene-OR ³ ;

R ⁶ is Cm-alkyl, OH, -OPO ₃ R ^3a 2; And

R ⁷ is H, Cm-alkyl, C _1-4 -alkylene-OH, C _1-4 -alkylene-OR ³ , where R ³ is as defined above, Cm-alkylene-E;

and where in option (iv) it is preferable

R ¹ represents H, NH2;

R ² represents H;

R ³ represents a group selected from H, -PO3R ^3a 2, -CH2-OPO3R ^3a 2 and -CH2-OC(=O)-R ^3b ;

R ^3a represents a hydrogen atom or cation suitable to form a pharmaceutically acceptable salt of either -CH2-OC(=O)-R ^3b or -CHMe-OC(=O)-R ^3b or -CMe2-OC(=O)-R ^3b or -CH2-OC(=O)-OR ^3b or -CHMe-OC(=O)-OR ^3b or -CMe2-OC(=O)-OR ^3b ;

R ^3b represents an alkyl group having from 1 to 11 carbon atoms, which is linear, branched or cyclic, or a combination thereof, wherein the R ^3b group is optionally substituted by one or more groups independently selected from -OH and -O-Cm-alkyl;

R ⁴ represents H, Cm-alkyl; And

R ⁵ represents H, C _1-4 -alkyl, C _1-4 -αalkylene-OR ³ ;

R ⁶ represents H, Cm-alkyl, OH, -OPO ₃ R ^3a 2; And

R ⁷ is Cm-alkyl, C _1-4 -alkylene-OH, C _1-4 -alkylene-OR ³ , where R ³ is as defined above, Cm-alkylene-E.

According to yet another aspect of this embodiment, the variable group Qi is a methylene group and Q2 has the same meaning as in the previous aspect, i.e. -CR ⁴ R ⁵ -CR ⁶ R ⁷ The compounds of this aspect are represented by the general formula VII

where R1, R ² , R ³ , R ⁴ , R ⁵ , R ⁶ and R ⁷ have the same meanings as defined for the general formula I above, which are defined in option (iii) or (iv) according to paragraphs. 1, 2 or 3 above, where in option (iii) it is preferable

R ¹ represents H, NH2;

R ² represents H;

R ³ represents a group selected from H, -PO3R ^3a 2, -CH ₂ -OPO ₃ R ^3a ₂ and -CH ₂ -OC(=O)-R ^3b ;

R ^3a represents a hydrogen atom or cation suitable to form a pharmaceutically acceptable salt of either -CH2-OC(=O)-R ^3b or -CHMe-OC(=O)-R ^3b or -CMe2-OC(=O)-R ^3b or -CH2-OC(=O)-OR ^3b or -CHMe-OC(=O)-OR ^3b or -CMe2-OC(=O)-OR ^3b ;

R ^3b represents an alkyl group having from 1 to 11 carbon atoms, which is linear, branched or cyclic, or a combination thereof, wherein the R ^3b group is optionally substituted by one or more groups independently selected from -OH and -O-Cm-alkyl;

R ⁴ represents H, C _1-4 -alkyl; And

R ⁵ represents H, C _1-4 -alkyl, C _1-4 -αalkylene-OR ³ ;

R ⁶ represents C1-4-alkyl, OH, -OPO3R ^3a 2; And

R ⁷ is H, C1-4-alkyl, C1-4-alkylene-OH, C1-4-alkylene-OR ³ where R ³ is as defined above, C1-4-alkylene-F; and where in option (iv) it is preferable

R ¹ represents H, NH2;

R ² represents H;

R ³ represents a group selected from H, -PO3R ^3a 2, -CH ₂ -OPO ₃ R ^3a ₂ and -CH ₂ -OC(=O)-R ^3b ;

R ^3a represents a hydrogen atom or cation suitable to form a pharmaceutically acceptable salt of either -CH2-OC(=O)-R ^3b or -CHMe-OC(=O)-R ^3b or -CMe2-OC(=O)-R ^3b or -CH2-OC(=O)-OR ^3b or -CHMe-OC(=O)-OR ^3b or -CMe2-OC(=O)-OR ^3b ;

R ^3b represents an alkyl group having from 1 to 11 carbon atoms, which is linear, branched or cyclic, or a combination thereof, wherein the R ^3b group is optionally substituted by one or more groups independently selected from -OH and -OC _1-6 -alkyl ;

- 20 046887

R ⁴ represents H, C _1-4 alkyl; And

R ⁵ represents H, C _1-4 alkyl, C ₁ . ₄ -αalkylene-OR ³ ;

R ⁶ represents H, C _1-4 -alkyl, OH, -OPO ₃ R ^3a 2; And

R ⁷ is C1.4-αalkyl, C1.4-αalkylene-OH, C1.4-alkylene-OR ³ , where R ³ is as defined above, C1-4-alkylene-F.

In another embodiment of the present invention, the bicyclic group on the left side of the molecule is linked through the 3-position of the five-membered heterocycle. The compounds of this embodiment are represented by the general formula VIII

where R ¹ , R ² , R ³ , Q1 , Q2 , Q ₃ have the same meanings as defined for the general formula I above.

Another preferred subtype of compounds of this aspect of the invention is represented by the general formula XI

R ¹

Wherein R ¹ , R ² , R ³ , R ⁴ , R ⁵ , R ⁶ and R ⁷ have the same meanings as defined for general formula I above, where in option (iii) it is preferable

R ¹ represents H;

R ² represents a group selected from H, -O-Ar ¹ , -O-Het ¹ , -NH2, where Ar ¹ represents a phenyl group which is optionally substituted by one or more groups selected from -CN, -O-C1-4 -αalkyl, or where the phenyl group may bear two substituent groups on adjacent ring atoms such that these adjacent substituent groups can be linked together to form a 5-membered heterocycle having one or two heteroatoms individually selected from N and O, where Het ¹ represents an aromatic heterocycle with 5 or 6 ring atoms including 1 or 2 heteroatoms individually selected from N, S and O, or a non-aromatic partially or fully saturated heterocycle with 6 ring atoms including 1 heteroatom selected from N, S and O, wherein the Het ¹ group may optionally be substituted by one or more groups individually selected from -CM-alkyl, -O-C1. ₄ -αalkyl, -CN;

R ³ represents a group selected from H, -PO3R ^3a 2, -CH2-OPO ₃ R ^3a ₂ and -CH ₂ -OC(=O)-R ^3b ;

R ^3a represents a hydrogen atom or cation suitable to form a pharmaceutically acceptable salt of either -CH2-OC(=O)-R ^3b or -CHMe-OC(=O)-R ^3b or -CMe2-OC(=O)-R ^3b or -CH2-OC(=O)-OR ^3b or -CHMe-OC(=O)-OR ^3b or -CMe2-OC(=O)-OR ^3b ;

R ^3b represents an alkyl group having from 1 to 11 carbon atoms, which is linear, branched or cyclic, or a combination thereof, wherein the R ^3b group may optionally be substituted by one or more groups independently selected from -OH and -O-Cj- 6-alkyl;

R ⁴ represents H, C ₁ . ₄ -alkyl; And

R ⁵ represents H, C ₁ . ₄ -alkyl, C ₁ . ₄ -alkylene-OR ³ .

R ⁶ represents C ₁ . ₄ -alkyl, OH, -OPO ₃ R ^3a 2; And

R ⁷ is H, C1.4-alkyl, C1.4-alkylene-OH, C1.4-alkylene-OR ³ where R ³ is as defined above, C1. ₄ -alkylene-F;

where in option (iv) it is preferable

R ¹ represents H;

R ² represents a group selected from H, -O-Ar ¹ , -O-Het ¹ , -NH2, where Ar ¹ represents a phenyl group which is optionally substituted by one or more groups selected from -CN, -OC ₁ . ₄ -alkyl, or where the phenyl group may bear two substituent groups on adjacent ring atoms such that these adjacent substituent groups can be bonded together to form a 5-membered heterocycle having one or two heteroatoms individually selected from N and O, where Het ¹ represents an aromatic heterocycle with 5 or 6 ring atoms comprising 1 or 2 heteroatoms individually selected from N, S and O, or a non-aromatic partially or fully saturated heterocycle with 6 ring atoms including 1 heteroatom selected from N, S and O where Het ¹ group may optionally be replaced by one or more groups individually selected from

- 21 046887

-C1. ₄ -alkyl, -O-C1. ₄ -alkyl, -CN;

R ³ represents a group selected from H, -PO3R ^3a 2, -CH2-OPO3R ^3a 2 and -CH2-OC(=O)-R ^3b ;

R ^3a represents a hydrogen atom or cation suitable to form a pharmaceutically acceptable salt of either -CH2-OC(=O)-R ^3b or -CHMe-OC(=O)-R ^3b or -CMe2-OC(=O)-R ^3b or -CH ₂ -OC(=O)-OR ^3b or -CHMe-OC(=O)-OR ^3b or -CMe2-OC(=O)-OR ^3b ;

R ^3b represents an alkyl group having from 1 to 11 carbon atoms, which is linear, branched or cyclic, or a combination thereof, wherein the R ^3b group is optionally substituted by one or more groups independently selected from -OH and -OC ₁ . ₆ -alkyl;

R ⁴ represents H, C ₁ . ₄ -alkyl; And

R ⁵ represents H, C ₁ . ₄ -alkyl, C ₁ . ₄ -alkylene-OR ³ .

R ⁶ represents H, C ₁ . ₄ -alkyl, OH, -OPO ₃ R ^3a 2; And

R ⁷ is C1.4-alkyl, C1.4-alkylene-OH, C1.4-alkylene-OR ³ , where R ³ is as defined above, C1. ₄ -alkylene-F.

Another preferred subtype of compounds of this aspect of the invention is represented by the general formula XII

R ¹

where R ¹ , R ² , R ³ , R ⁶ and R ⁷ have the same meanings as defined for general formula I above, where in option (iii) it is preferable

R ¹ represents H;

R ² represents a group selected from H, -O-Ar ¹ , -O-Het ¹ , -NH2, where Ar ¹ represents a phenyl group which is optionally substituted by one or more groups selected from -CN, -OC _1-4 - alkyl, or wherein the phenyl group may bear two substituent groups on adjacent ring atoms such that these adjacent substituent groups may be bonded together to form a 5-membered heterocycle having one or two heteroatoms individually selected from N and O, where Het ¹ represents an aromatic heterocycle with 5 or 6 ring atoms comprising 1 or 2 heteroatoms individually selected from N, S and O, or a non-aromatic partially or fully saturated heterocycle with 6 ring atoms comprising 1 heteroatom selected from N, S and O, where Het ¹ group is optionally substituted with one or more groups individually selected from ^^-alkyl, -O-CM-alkyl, -CN;

R ³ represents a group selected from H, -PO3R ^3a 2, -CH ₂ -OPO ₃ R ^3a ₂ and -CH ₂ -OC(=O)-R ^3b ;

R ^3a represents a hydrogen atom or cation suitable to form a pharmaceutically acceptable salt of either -CH2-OC(=O)-R ^3b or -CHMe-OC(=O)-R ^3b or -CMe2-OC(=O)-R ^3b or -CH2-OC(=O)-OR ^3b or -CHMe-OC(=O)-OR ^3b or -CMe2-OC(=O)-OR ^3b ;

R ^3b represents an alkyl group having from 1 to 11 carbon atoms, which is linear, branched or cyclic, or a combination thereof, wherein the R ^3b group is optionally substituted by one or more groups independently selected from -OH and -OC _1-6 -alkyl ;

R ⁴ represents H, C _1-4 -alkyl; And

R ⁵ represents H, C _1-4 -alkyl, C _1-4 -alkylene-OR ³ ;

R ⁶ represents C1-4-alkyl, OH, -OPO3R ^3a 2; And

R ⁷ represents H, C1-4-alkyl, C1-4-alkylene-OH, C1-4-alkylene-OR ³ , where R ³ is as defined above, C1-4-alkylene-F;

where in option (iv) it is preferable

R ¹ represents H;

R ² represents a group selected from H, -O-Ar ¹ , -O-Het ¹ , -NH2, where Ar ¹ represents a phenyl group which may be optionally substituted by one or more groups selected from -CN, -OC _{1- 4} -alkyl, or where the phenyl group may bear two substituent groups on adjacent ring atoms such that these adjacent substituent groups can be bonded together to form a 5-membered heterocycle having one or two heteroatoms individually selected from N and O, where Het ¹ represents an aromatic heterocycle with 5 or 6 ring atoms comprising 1 or 2 heteroatoms individually selected from N, S and O, or a non-aromatic partially or fully saturated heterocycle with 6 ring atoms including 1 heteroatom selected from N, S and O wherein the Het ¹ group is optionally substituted with one or more groups individually selected from ^^-alkyl, -O-CM-alkyl, -CN;

R ³ represents a group selected from H, -PO3R ^3a 2, -CH ₂ -OPO ₃ R ^3a ₂ and -CH ₂ -OC(=O)-R ^3b ;

R ^3a represents a hydrogen atom or cation suitable to form a pharmaceutically acceptable salt of either -CH2-OC(=O)-R ^3b or -CHMe-OC(=O)-R ^3b or -CMe2-OC(=O)-R ^3b or -CH2-OC(=O)-O

- 22 046887

R ^3b or -CHMe-OC(=O)-OR ^3b or -CMe2-OC(=O)-OR ^3b ;

R ^3b represents an alkyl group having from 1 to 11 carbon atoms, which is linear, branched or cyclic, or a combination thereof, wherein the R ^3b group is optionally substituted by one or more groups independently selected from -OH and -OC _1-6 -αalkyl ;

R ⁴ represents H,C^-alkyl; And

R ⁵ represents H, O^-alkyl, C _1-4 -alkylene-OR ³ .

R ⁶ represents H, Ohm-alkyl, OH, -OPO ₃ R ^3a 2; And

R ⁷ is Q-4-alkyl, C1-4-alkylene-OH, C1-4-alkylene-OR ³ , where R ³ is as defined above, C1-4-alkylene-F.

Another embodiment of the present invention relates to compounds represented by the general formula XIII

ABOUT

where R ³ , R ⁸ , Q1, Q2 have the same meanings as defined for the general formula I above.

In another preferred aspect of this embodiment, an NH amino group is present at the Q ₁ position and a -CR ⁴ R ⁵ -CR ⁶ R ⁷ group is present at the Q2 position. The compounds are represented by the general formula XV

where R ³ , R ⁴ , R ⁵ , R ⁶ , R ⁷ , R ⁸ have the same meanings as defined for general formula I above, where in option (iii) it is preferable

R ³ represents a group selected from H, -PO3R ^3a 2, -CH ₂ -OPO ₃ R ^3a ₂ and -CH ₂ -OC(=O)-R ^3b ;

R ^3a represents a hydrogen atom or cation suitable to form a pharmaceutically acceptable salt of either -CH2-OC(=O)-R ^3b or -CHMe-OC(=O)-R ^3b or -CMe2-OC(=O)-R ^3b or -CH2-OC(=O)-OR ^3b or -CHMe-OC(=O)-OR ^3b or -CMe2-OC(=O)-OR ^3b ;

R ^3b represents an alkyl group having from 1 to 11 carbon atoms, which is linear, branched or cyclic, or a combination thereof, wherein the R ^3b group is optionally substituted by one or more groups independently selected from -OH and ^^^-alkyl;

R ⁴ represents H, O^-alkyl; And

R ⁵ represents H, Ohm-alkyl, C _1-4 -alkylene-OR ³ ;

R ⁶ represents C _1-4 -alkyl, OH, -OPO ₃ R ^3a 2; And

R ⁷ is H, O^-alkyl, C1-4-alkylene-OH, C1-4-alkylene-OR ³ , where R ³ is as defined above, C1-4-alkylene-F; And

R ⁸ represents a group selected from -O-Ar ² and -O-Het ² , where Ar ² represents a phenyl group that is optionally substituted by one or more groups selected from -CN, -O-CM-alkyl, or where the phenyl group may bear two substituent groups on adjacent ring atoms such that these adjacent substituent groups can be linked together to form a 5-membered heterocycle having one or two heteroatoms individually selected from N, S and O, wherein said heterocycle may optionally bear one or two substituent selected from oxo, F, -OC _1-4 -alkyl, C _1-4 -alkyl, CN; and where Het ² represents an aromatic heterocycle with 5 or 6 ring atoms, including 1 or 2 heteroatoms individually selected from N, S and O, which are optionally substituted by one or more groups individually selected from -OC _1-4 -alkyl, -CN , F, C _1-4 -alkyl;

where in option (iv) it is preferable

R ³ represents a group selected from H, -PO3R ^3a 2, -CH ₂ -OPO ₃ R ^3a ₂ and -CH ₂ -OC(=O)-R ^3b ;

R ^3a represents a hydrogen atom or cation suitable to form a pharmaceutically acceptable salt of either -CH2-OC(=O)-R ^3b or -CHMe-OC(=O)-R ^3b or -CMe2-OC(=O)-R ^3b or -CH2-OC(=O)-OR ^3b or -CHMe-OC(=O)-OR ^3b or -CMe2-OC(=O)-OR ^3b ;

R ^3b represents an alkyl group having from 1 to 11 carbon atoms, which is linear

- 23 046887 ny, branched or cyclic, or a combination thereof, where the R ^3b group is optionally substituted by one or more groups independently selected from -OH and _{-OC 1_6 -alkyl} ;

R ⁴ represents H, Cm-alkyl; And

R ⁵ represents H, C _1-4 alkyl, C ₁ . ₄ -alkylene-OR ³ ;

R ⁶ represents H, C _1-4 -alkyl, OH, -OPO ₃ R ^3a 2; And

R ⁷ is C1-4-alkyl, C1-4-alkylene-OH, C1-4-alkylene-OR ³ where R ³ is as defined above, C1-4-alkylene-F; And

R ⁸ represents a group selected from -O-Ar ² and -O-Het ² , where Ar ² represents a phenyl group that is optionally substituted by one or more groups selected from -CN, -O-C _1-4 -alkyl, or wherein the phenyl group may bear two substituent groups on adjacent ring atoms such that these adjacent substituent groups may be bonded together to form a 5-membered heterocycle having one or two heteroatoms individually selected from N, S and O, wherein said heterocycle may optionally bear one or two substituents selected from oxo, F, -OC _1-4 -alkyl, C _1-4 -alkyl, CN; and where Het ² represents an aromatic heterocycle with 5 or 6 ring atoms, including 1 or 2 heteroatoms individually selected from N, S and O, which may optionally be substituted by one or more groups individually selected from -OC _1-4 -alkyl, -CN, F, C _1-4 -alkyl.

In a related aspect, the variable group Q1 represents a methylene group such that the compounds are represented by the general formula XVI

where R ³ , R ⁴ , R ⁵ , R ⁶ , R ⁷ , R ⁸ have the same meanings as defined for general formula I above, where in option (iii) it is preferable

R ³ represents a group selected from H, -PO3R ^3a 2, -CH^OPCm ³ ^ and -CH ₂ -OC(=O)-R ^3b ;

R ^3a represents a hydrogen atom or cation suitable to form a pharmaceutically acceptable salt of either -CH2-OC(=O)-R ^3b or -CHMe-OC(=O)-R ^3b or -CMe2-OC(=O)-R ^3b or -CH2-O-C(=O)-OR ^3b or -CHMe-OC(=O)-OR ^3b or -CMe2-OC(=O)-OR ^3b ;

R ^3b represents an alkyl group having from 1 to 11 carbon atoms, which is linear, branched or cyclic, or a combination thereof, wherein the R ^3b group is optionally substituted by one or more groups independently selected from -OH and -O-C _1-6 -alkyl;

R ⁴ is H, C _1-4 alkyl; And

R ⁵ is H, C _1-4 -alkyl, C _1-4 -alkylene-OR ³ ;

R ⁶ represents C1-4-alkyl, OH, -OPO3R ^3a 2; And

R ⁷ is H, C1-4-alkyl, C1-4-alkylene-OH, C1-4-alkylene-OR ³ , where R ³ is as defined above, C1-4-alkylene-P;

R ⁸ represents a group selected from -O-Ar ² and -O-Het ² , where Ar ² represents a phenyl group which may be optionally substituted by one or more groups selected from -CN, -O-C _1-4 -alkyl , or where the phenyl group may bear two substituent groups on adjacent ring atoms such that these adjacent substituent groups may be bonded together to form a 5-membered heterocycle having one or two heteroatoms selected from N, S and O, wherein said heterocycle may optionally carry one or two substituents individually selected from oxo, F, -O-C _1-4 -alkyl, C _1-4 -alkyl, CN; and -y where Het represents an aromatic heterocycle with 5 or 6 ring atoms, including 1 or 2 heteroatoms individually selected from N, S and O, which are optionally substituted by one or more groups individually selected from -O-C _1-4 -alkyl , -CN, F, C _1-4 -alkyl;

where in (iv) it is preferable

R ³ represents a group selected from H, -PO ₃ R ^3a 2, -CH2-OPO3R ^3a 2 and -CH ₂ -OC(=O)-R ^3b ;

R ^3a represents a hydrogen atom or cation suitable to form a pharmaceutically acceptable salt of either -CH2-OC(=O)-R ^3b or -CHMe-OC(=O)-R ^3b or -CMe2-OC(=O)-R ^3b or -CH2-O-C(=O)-OR ^3b or -CHMe-OC(=O)-OR ^3b or -CMe2-OC(=O)-OR ^3b ;

R ^3b represents an alkyl group having from 1 to 11 carbon atoms, which is linear, branched or cyclic, or a combination thereof, wherein the R ^3b group is optionally substituted by one or more groups independently selected from -OH and -O-C _1-6 -alkyl;

R ⁴ is H, C _1-4 alkyl; And

R ⁵ is H, C _1-4 -alkyl, C _1-4 -alkylene-OR ³ ;

- 24 046887

R ⁶ represents H, C _1-4 -alkyl, OH, -OPO3R ^3a 2; And

R ⁷ represents C1-4-alkyl, C1-4-alkylene-OH, C.'|-4-alkylene-OK ³ , where R ³ is as defined above, C1-4-alkylene-G;

R ⁸ represents a group selected from -O-Ar ² and -O-Het ² , where Ar ² represents a phenyl group that is optionally substituted by one or more groups selected from -CN, -OC _1-4 -alkyl, or where phenyl group may bear two substituent groups on adjacent ring atoms such that these adjacent substituent groups can be linked together to form a 5-membered heterocycle having one or two heteroatoms selected from N, S and O, wherein said heterocycle may optionally bear one or two substituents individually selected from oxo, F, -OC _1-4 -alkyl, ^^-alkyl, CN; and where Het ² represents an aromatic heterocycle with 5 or 6 ring atoms, comprising 1 or 2 heteroatoms individually selected from N, S and O, which may optionally be substituted by one or more groups individually selected from -OQ-4-alkyl, - CN, F, C _1-4 -alkyl.

As used herein, all such compounds, including cis- and trans-isomers, R- and S-enantiomers, diastereomers, (1)-isomers, racemic mixtures thereof and other mixtures thereof, are within the scope of the invention. However, the carbon double bond The -carbon between the pyridine ring and the amide group at the center of the molecule must be in the trans configuration, as shown in the above formulas. Additional asymmetric carbon atoms may be present in a substituent such as an alkyl group. All such isomers, as well as mixtures thereof, are intended to be included. into the present invention.

If, for example, a particular enantiomer of a compound described herein is desired, it can be prepared by asymmetric synthesis or derivatization with a chiral auxiliary where the resulting diastereomeric mixture is separated and the auxiliary group is cleaved off to obtain the pure desired enantiomers. Alternatively, when the molecule contains a basic functional group, such as amino, or an acidic functional group, such as carboxyl, diastereomeric salts are formed with the appropriate optically active acid or base, followed by separation of the diastereomers thus formed by means of fractional crystallization or chromatography well known in the art. this field of technology, and the subsequent recovery of pure enantiomers.

Moreover, the individual enantiomers and diastereomers of the compounds of the present invention can be prepared synthetically from commercially available starting materials that contain asymmetric or stereogenic centers, or by preparing racemic mixtures followed by resolution by methods well known to those skilled in the art. These separation methods include: (1) addition of a mixture of enantiomers to a chiral auxiliary, separation of the resulting mixture of diastereomers by recrystallization or chromatography and isolation of the optically pure product from the auxiliary, (2) formation of a salt using an optically active resolving agent, (3) direct separation of the mixture optical enantiomers on chiral liquid chromatography columns or (4) kinetic separation using stereoselective chemical or enzymatic reagents. Racemic mixtures can also be separated into their constituent enantiomers by well-known methods such as chiral gas-phase chromatography or crystallization of the compound in a chiral solvent. Stereoselective synthesis, a chemical or enzymatic reaction in which one reagent forms an unequal mixture of stereoisomers during the creation of a new stereocenter or during the transformation of an existing one, is well known in the art. Stereoselective synthesis involves both enantio- and diastereoselective transformations. For examples, see Carreira and Kvaerno, Classics in Stereoselective Synthesis, Wiley-VCH: Weinheim, 2009.

The compounds described herein may exist in solvated as well as non-solvated forms with pharmaceutically acceptable solvents such as water, ethanol and the like, and it is intended that the invention covers both solvated and non-solvated forms. The compounds described herein may exist in one or more crystalline forms or polymorphs. In one embodiment, the compound is amorphous. In one embodiment, the compound is a single polymorph. In another embodiment, the compound is a mixture of polymorphs. In another embodiment, the compound is in crystalline form.

The invention also covers the isotopically labeled compounds of the invention as provided herein, except that one or more atoms are replaced by an atom having an atomic mass or mass number different from the atomic mass or mass number typically found in nature. Examples of isotopes that may be included in the compounds of the invention include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorus, fluorine and chlorine, such as ² H, ³ H, ¹³ C, ¹⁴ C, ¹⁵ N, ¹⁸ O, ¹⁷ O , ³¹ P, ³² P, ³⁵ S, ¹⁸ F, and ³⁶ Cl, respectively. For example, a compound of the invention may have one or more H atoms substituted with deuterium.

Some described isotopically labeled compounds (e.g. labeled with 3H and ^14C ) are used in

- 25 046887 analyzes of the distribution of the compound and/or substrate in tissues. Tertiated (ie ^3H ) and carbon-14 (ie ^14C ) isotopes are particularly preferred due to their ease of preparation and detection. Additionally, substitution with heavier isotopes such as deuterium (i.e., ^2H ) may provide certain therapeutic benefits due to greater metabolic stability (e.g., increased in vivo half-life or reduced dosage requirements) and may therefore be preferable in some circumstances. The isotopically labeled compounds of the invention can generally be prepared using the following methods, similar to those described in, for example, the examples herein, through the substitution of an isotopically labeled reagent for a non-isotopically labeled reagent.

The prodrugs of the present invention contain at least one prodrug group, i.e. a group that is cleaved under physiological conditions, thereby releasing active species. Such prodrug groups can be attached to the compounds of the present invention at all positions represented by the R ³ variable group. A suitable prodrug group is the methylene phosphate group as described in WO 2013/190384 A1. Other suitable prodrug groups are phosphates or other solubilizing groups, as described in Prodrugs: design and clinical applications (Rautio et al. Nature Reviews Drug Discovery, 2008, 7, 255). One specific modification of a prodrug is the conversion of an amino group to an N-oxide group. The above chemical formulas illustrate suitable provisions for modification of the prodrug with a methylene phosphate group and a phosphate group. It should be understood that these prodrug groups may be replaced by other prodrug groups. According to a preferred embodiment of the present invention, the prodrugs of the present invention contain exactly one prodrug group.

Production methods.

The compounds of the present invention can be prepared using known organic chemistry methods and procedures and/or the information described herein below. Starting materials can either be purchased (if commercially available) or can be synthesized using conventional organic chemistry methods and procedures and/or the information described herein below.

Final amide coupling step.

The compounds described herein can be prepared by the following method, which includes the step of coupling a precursor compound of formula XVII

wherein Q1 and Q2 have the same meanings as defined herein above, wherein R ³ represents a hydrogen atom and where X represents a leaving group, with an amine compound of formula H-NA1A2, sometimes referred to herein as formula XXIII, wherein A is ₁ and A ₂ has the same meaning as defined herein above for the compound of Formula I.

The leaving group can be selected from hydroxyl group, tosylate group, triflate group, mesylate group, iodide, bromide and the like.

Especially if the leaving group is a hydroxyl group, the coupling reaction is usually carried out in a solvent and in the presence of a coupling agent and, more preferably, in the presence of a solvent, coupling groups and a base. The solvent is preferably selected from DMF, THF, DCM. The coupling agent is preferably selected from GATU, GBTU, GCTU, TBTU, COMU, TOMBU, COMBU and, most preferably, GATU. The reaction is usually carried out in the presence of a base. The base is preferably selected from DIPEA.

The sequence of this reaction is illustrated in the following Scheme 1.

Scheme 1

where A1 and A2 as well as Q1 and Q2 have the same meanings as defined for compound I herein above, and where R ³ represents a hydrogen atom.

Production of the predecessor of the right side.

The precursor compound of formula XVII can be produced by reacting the compound

- 26 046887 formula XVIII

wherein Q ₁ and Q ₂ have the same meanings as defined for compound I herein above, and where R ³ represents a hydrogen atom, with a carboxyl-protected acrylic acid such as a C _1-6 alkyl ester (preferably tert- butyl ester) of aryl acid. This coupling reaction is carried out under Heck coupling conditions and preferably in the presence of a Pd(II) salt such as Pd(OAc)2 and a phosphine ligand such as Xantphos, tri-(o-tolyl)phosphine or 1,1-bis (diphenylphosphino)ferrocene(dppf). The reaction is usually carried out in the presence of a solvent such as DMF propionitrile or a combination thereof, and also in the presence of a base such as DIPEA. The coupling reaction is carried out through deprotection and optional introduction of a leaving group other than hydroxyl. This reaction sequence is illustrated in the following Scheme 2.

Scheme 2

where Pg represents a protecting group, Q1 and Q2 have the same meanings as defined for compound I herein above, and where R ³ represents a hydrogen atom.

Precursor compound XX can be synthesized using a cyclization reaction starting from the compound of formula XXI

where Q ₁ and Q ₂ have the same meanings as defined for compound I herein above, and where R ³ represents a hydrogen atom. Cyclization is usually carried out using coupling agents such as GATU, in the presence of a base such as DIPEA and in a solvent such as DMF. This reaction sequence is illustrated in the following Scheme 3.

Scheme 3

meanings as defined herein above, and where R ³ where Q1 and Q2 are the same represents a hydrogen atom.

Alternatively, the precursor compound XX can be synthesized using a cyclization reaction starting with the compound of formula XXII

where Q1 and Q2 have the same meanings as defined for compound I herein

- 27 046887 above.

This variant of the cyclization reaction is usually carried out in the presence of a reducing agent such as Fe in acetic acid or a mixture of water, ethanol and ammonium chloride. This reaction step is illustrated by the following reaction scheme 4.

Scheme 4

where Q1 and Q2 have the same meanings as defined for compound I herein above, and where R ³ represents a hydrogen atom.

Production of H-NA1A2 precursor.

If the variable group A1 is a group selected from groups A11 and A12, and A2 is a methyl group, the precursor compound H-NA1A2 can be prepared using the following procedures.

One option is to form the precursor amide XXIV, which is subsequently reduced to amine compound XXIII. The amide precursor XXIV has the following structure (where the XXIV structure allows an amide group to be bonded to position 2, and a methyl group bonded to position 3, or vice versa)

wherein R ¹ , R ² and Q ₃ have the same meaning as defined for compound I above, or where R ¹ and/or R ² may represent a functional group that can be converted to the desired substituent at a subsequent step in the reaction sequence. Said functional group must, of course, be inert under the reaction conditions defined for the transformations described herein (ie, from compound XXV to compound XXIII), but be susceptible to subsequent conversion under other reaction conditions. It could be, for example, a Br substituent.

Compound XXIV can be synthesized by reacting the activated carboxylic acid derivative XXV

with methylamine. In formula XXV, the variable group X represents a leaving group, which may be, for example, an alkoxy group, -OHet, such as 1-hydroxybenzotriazole, carbamimidate, while R ¹ , R ² and Q ₃ have the same meaning as indicated for the compound I above.

In the next step, the amide group of compound XXIV is reduced to give amine XXIII. This can be accomplished using a conventional reducing agent suitable for reducing carbonyl groups, such as BH ₃ . This reaction is usually carried out in a solvent such as THF.

The sequence of reaction steps described above is illustrated by the following reaction scheme 5.

Scheme 5

Alternatively, amine compound XXIII can be prepared from the corresponding derivative of formula XXVI

- 28 046887

wherein R ¹ , R ² and Q3 have the same meaning as defined for compound I above, or where R ¹ and/or R ² may represent a functional group that can be converted to the desired substituent in a subsequent step in the reaction sequence as described above .

The compound of formula XXVI is reacted with a protected N-methylamine in the presence of a borohydride compound such as Na(OAc)3BH in the presence of a solvent such as ethanol or 1,2dichloroethane. Suitable protecting groups are benzyl or p-methoxybenzyl. Intermediate XXVII is obtained.

In formula XXVII, Pg represents a protecting group and R ¹ , R ² and Q3 have the same meaning as given for compound I above, or R ¹ and/or R ² may represent a functional group that can be converted to the desired substituent at a later time reaction sequence steps as described above.

After the reaction, the protecting group is removed, usually by hydrogenolysis, to yield amine compound XXIII. The sequence of reactions is illustrated in the following Scheme 6.

Scheme 6

together form group A3, precursor compound XXIII

If the variables of group A1 and _A2 have the following structure XXIII:

where R ⁸ has the same meaning as defined for compound I above.

Compound XXIIIa can be prepared by combining the precursor compound XXIV

where R ⁸ has the same meaning as defined for compound I above, with the precursor compound XXV

followed by deprotection according to the following reaction scheme 7. The coupling reaction is usually carried out in the presence of a carbonate, such as Na ₂ CO3 or K2CO3, and a Pd complex, which is under

- 29 046887 is conveniently selected from Pd(PPh3) ₄ and PdCl2(dppf). A suitable reaction solvent is a mixture of water and 1,4-dioxane. A preferred protecting group is the BOC group.

Scheme 7

where Pg represents a protecting group such as Boc, and R ⁸ has the same meaning as defined for compound I above.

Alternatively, the target compound XXIIIa can be prepared by combining the precursor compound XXVI

with precursor compound XXVII after deprotection according to the following reaction scheme 8. The coupling reaction is usually carried out in the presence of a carbonate such as K2CO3 and a Pd complex such as PdCl2(dppf). A suitable reaction solvent is 1,4-dioxane or a mixture of water and 1,4-dioxane. A preferred protecting group is the BOC group.

Scheme 8

In the reaction scheme above, Pg represents a protecting group and R ⁸ is as defined for compound I above.

- 30 046887

The final stage is a combination of Hake.

According to an alternative synthesis strategy, the compounds described herein can be produced using the following method, which includes the step of coupling a precursor compound of formula XVIII.

wherein Q1 and Q2 have the same meanings as defined for compound I herein above, and wherein ^R3 represents a hydrogen atom, with the precursor compound of formula XIX

where A ₁ and A ₂ have the same meanings as defined herein above.

This coupling reaction is carried out under Heck coupling conditions. It is typically carried out in the presence of a Pd(II) complex such as Pd-162 (ie [P(tBu) ₃ ]Pd(crotyl)Cl), tetrabutylammonium chloride, Ncyclohexyl-N-methylcyclohexanamine and dioxane. It is also possible to use a combination of a Pd(II) salt, such as Pd(OAc)2, with a phosphine ligand, such as tri-o-tolylphosphane, a base, such as DIPEA, and a solvent, such as a mixture of DMF and ACN. The reaction is illustrated by the following reaction scheme. 9.

Scheme 9

In the above reaction scheme, Q1, Q2, A1 and A2 have the same meanings as defined for compound I above, and R ³ represents hydrogen.

Preparation of prodrugs of compounds of the present invention is generally accomplished by converting the corresponding compound of the present invention, where R ³ is hydrogen, into a compound with the same structure, except that R ³ is a prodrug group that is cleaved under physiological conditions, such as a phosphate-containing group , as mentioned above. The prodrug group is usually a methylene phosphate group. Such prodrug groups and suitable reaction conditions for the preparation of methylene phosphate prodrugs are described in WO 2013/190384 A1.

Alternatively, the prodrug group may be linked to other parts of the molecule. In particular, it is considered to bind to the prodrug group on the right side of the molecule through the variable groups R ⁵ or R ⁷ , in which case the variable groups are a C _1-4 -alkyleneOR ³ group. The method for preparing such prodrugs is similar to the method described in WO 2013/190384 A1, but where the nitrogen atom bearing R ³ must of course be suitably protected.

Pharmaceutical compositions.

Forms of administration, pharmaceutical compositions and formulations.

The pharmaceutical compositions herein may be administered in a variety of ways depending on their intended use, as is well known in the art. For example, if the compositions disclosed are to be administered orally, they may be formulated as tablets, capsules, granules, powders or syrups. Alternatively, the compositions described herein may be administered parenterally as injections (intravenous, intramuscular, or subcutaneous), drips, or suppositories. For use across the ocular mucosa, the compositions described herein may be formulated as eye drops or ophthalmic ointments. These compositions can be prepared by conventional methods and, if desired, the compositions can be mixed with any conventional additive such as an excipient, a binding agent, a disintegrant, a lubricant, a flavoring agent, a solubilizing agent, a suspension aid, an emulsifying agent or a coating agent. The excipients described may perform more than one function. For example, fillers or binding agents may also be disintegrating agents, glidants, antiblocking agents, lubricants, sweeteners, and the like.

In the formulations described, wetting agents, emulsifiers and lubricants such as sodium lauryl sulfate and magnesium stearate, as well as colorants, release agents, coating agents, sweeteners, flavoring agents, preservatives and antioxidants may be present in the formulated agents.

- 31 046887

Subject compositions may be suitable for oral, nasal (eg, inhalation using a dry powder or nebulized composition), topical (including buccal and sublingual), pulmonary (including aerosol), rectal, vaginal, aerosol and/or parenteral ( for example, by injection, for example, intravenous, intramuscular or subcutaneous injection) administration. The compositions may be conveniently presented in unit dosage form, and may be prepared by any methods well known in the art of pharmacy. The amount of a compound described herein that can be combined with a carrier material to produce a single dose may vary depending on the identity of the compound, the subject being treated, and the particular route of administration.

Methods for preparing these compositions include the step of combining the compositions as described with a carrier and, optionally, one or more additional ingredients. Typically, formulations are prepared by uniformly and thoroughly combining the agents with liquid carriers or finely divided solid carriers, or both, and then, if necessary, molding the product.

Formulations suitable for oral administration may be in the form of capsules, wafers, pills, tablets, lozenges (using a flavored base, usually sucrose and gum acacia or tragacanth), powders, granules, or as a solution or suspension in an aqueous or non-aqueous liquid , or in the form of a liquid emulsion of oil in water or water in oil, or in the form of an elixir or syrup, or in the form of lozenges (using an inert base such as gelatin and glycerin, or sucrose and acacia), each containing a predetermined amount the composition in question as an active ingredient. The compositions herein may also be administered as a bolus, electuary, or paste.

In solid dosage forms for oral administration (capsules, tablets, pills, dragees, powders, granules and the like), the subject composition is mixed with one or more pharmaceutically acceptable excipients selected from: (1) fillers or extenders such as starches, dextrose, lactose , sucrose, glucose, mannitol and/or silicic acid; (2) binders such as, for example, cellulose (eg, microcrystalline cellulose, methylcellulose, hydroxypropylmethylcellulose (HPMC) and carboxymethylcellulose), alginates, gelatin, polyvinylpyrrolidone, sucrose and/or gum acacia; (3) humectants such as glycerin; (4) disintegrants such as croscarmellose sodium, sodium carboxymethyl starch (sodium starch glycolate), cross-linked polyvinyl purrolidone (crospovidone), gellan gum, xanthan gum, agaragar, calcium carbonate, potato or tapioca starch, alginic acid and sodium alginate, certain silicates and especially calcium silicate and sodium carbonate; (5) dissolution retarding agents such as paraffin; (6) absorption accelerators such as quaternary ammonium compounds; (7) wetting agents such as, for example, cetyl alcohol and glycerol monostearate; (8) absorbents such as kaolin and bentonite clay; (9) lubricants such as talc, calcium stearate, magnesium stearate, solid polyethylene glycols, sodium lauryl sulfate and mixtures thereof; (10) dyes; (11) complexing agents such as cyclodextrins and modified cyclodextrins, including simple (2-hydroxypropyl)-b-cyclodextrin and sulfobutyl ether-b-cyclodextrin; (12) matrix-forming polymer fillers such as polyvinylpyrrolidine (PVP), for example PVP K30, acrylic polymers and copolymers, such as various grades of Eudragit and, preferably, Eudragit L100, hydroxypropyl methylcellulose succinate (HPMCAS), other copolymers, such as copolymers based on polyethylene glycol, such as Soluplus; and (13) carriers such as sodium citrate or dicalcium phosphate. In the case of capsules, tablets and pills, the compositions may also contain buffering agents. Solid compositions of this type can also be used as fillers in soft and hard gelatin capsules using fillers such as lactose or milk sugars, as well as high molecular weight polyethylene glycols and the like. The excipients described may perform more than one function. For example, fillers or binding agents may also be disintegrating agents, glidants, antiblocking agents, lubricants, sweeteners, and the like. In accordance with the present invention, two or more excipients can be used, where the two or more excipients may belong to the same and/or different categories. There are no restrictions in this regard.

Preferred oral formulations.

Binding agents.

A coupling agent is primarily used to increase the particle size of a single active ingredient or with excipients and improve its performance properties. There are no particular restrictions on the binder material that can be used in the present invention.

Suitable binders include povidone (polyvinylpyrrolidone), copovidone (copolymer of poly(1-vinylpyrrolidone-co-vinyl acetate)), maltodextrin, poloxamer (a block copolymer with a first block of poly(ethylene oxide), a second and central block of poly(propylene oxide) and a third block of poly( ethylene oxide)), polyethylene glycol, polyethylene oxide, magnesium aluminum silicate, gelatin, gum arabic, alginic acid, carbomer (carbopol), dextrin, dextrates (purified sugar mixture

- 32 046887 reads, obtained by controlled enzymatic hydrolysis of starch), guar gum, hydrogenated vegetable oil, liquid glucose, wax, starch (pregelatinized and plain), sodium alginate and mixtures thereof.

The use of povidone and copovidone is preferred.

The coupling agent may be present in a relative amount of 0.5 wt% to 15 wt%, preferably 1 wt% to 12 wt%, and more preferably 4 wt% to 10 wt%.

Thinners.

Diluents are advantageously used to increase the volume of a pharmaceutical composition and to facilitate handling of the composition. There are no particular restrictions on the diluent material that can be used in the present invention.

Suitable diluents include mannitol, isomalt, histidine, lactose (including anhydrous or monohydrate forms), calcium phosphate (including dibasic and tribasic calcium phosphate), calcium carbonate, calcium sulfate, sucrose, fructose, maltose, xylitol, sorbitol, maltitol, aluminum silicate , dextrose, starch (pregelatinized or plain), glucose, dextrates (a purified mixture of saccharides obtained by controlled enzymatic hydrolysis of starch), magnesium carbonate and mixtures thereof.

Preferably, mannitol, xylitol, sorbitol, isomalt and/or histidine are used. Mannitol is particularly preferred.

The diluent may be present in a relative amount that is not particularly limited. Suitable amounts may range from 2% by weight to 85% by weight, preferably from 8% by weight to 80% by weight, and more preferably from 10% by weight to 50% by weight.

Surface-active substance.

The surfactant may advantageously be used to improve the wettability of the tablet and the active ingredient. A surfactant is an optional but preferred component. There are no particular restrictions on the surfactant that can be used in the present invention.

Suitable surfactants include sodium lauryl sulfate, poloxamer, sodium docusate, sorbitan esters, polyethylene oxide, polysorbate 20, polysorbate 40, polysorbate 60, polysorbate 80 (ethoxylated sorbitan esterified with fatty acids, where the number indicates the number of repeating units of polyethylene glycol) and their mixtures.

Preferably use sodium lauryl sulfate.

The surfactant may be present in a relative amount, which is not particularly limited. Suitable amounts may be from 0% by weight or more to 7% by weight, preferably from 0.1% by weight to 6.5% by weight and more preferably from 1% by weight to 6% by weight.

Baking powder.

Disintegrant is used to accelerate the breakdown of a pharmaceutical composition, thereby promoting dissolution and absorption of the active ingredient. There are no particular restrictions on the leavening agent that can be used in the present invention.

Suitable disintegrants include cross-linked polyvinylpyrrolidone (crospovidone), sodium carboxymethyl starch (sodium starch glycolate), croscarmellose sodium, gellan gum, xanthan gum, magnesium aluminum silicate, sodium alginate, pregelatinized starch, alginic acid, guar gum, homo- and copolymers (meth) acrylic acid and its salts, such as potassium polacrillin, and mixtures thereof.

The preferred use is crospovidone.

The disintegrant may be present in a relative amount that is not particularly limited. Suitable amounts may be from 0% by weight or more to 20% by weight, preferably from 1% by weight to 15% by weight, and more preferably from 2% by weight to 10% by weight.

Glidant.

Glidant is mainly used to improve the fluidity of a pharmaceutical composition, thereby improving its technological properties. Glidant is an optional but preferred component. There are no particular restrictions on the glidant material that can be used in the present invention.

Suitable glidant materials include colloidal silica, magnesium oxide, magnesium silicate, tribasic calcium phosphate, and mixtures thereof.

Preferably, colloidal silica is used.

The glidant may be present in a relative amount that is not particularly limited. Suitable amounts may be from 0% by weight or more to 5% by weight, preferably from 0.1% by weight to 4% by weight, and more preferably from 0.2% by weight to 1% by weight.

Lubricating agent.

Lubricants are advantageously used to facilitate tabletting, in particular by preventing tablets from sticking to the tablet punch. A lubricant is an optional but preferred component. There are no special restrictions on the material I lubricate

- 33 046887 common agent that can be used in the present invention.

Suitable lubricants include magnesium stearate, sodium stearyl fumarate, talc, stearic acid, leucine, poloxamer, polyethylene glycol, glyceryl behenate, glycerol monostearate, magnesium lauryl sulfate, sucrose fatty acid esters, calcium stearate, aluminum stearate, hydrogenated castor oil, hydrogenated vegetable oil, mineral oil, sodium benzoate, zinc stearate, palmitic acid, carnauba wax, sodium lauryl sulfate, polyoxyethylene monostearates, calcium silicate and mixtures thereof.

The use of a lubricant selected from magnesium stearate and sodium stearyl fumarate, and combinations thereof, is preferred.

The lubricating agent may be present in a relative amount that is not particularly limited. Suitable amounts may be from 0% by weight or more to 7% by weight, preferably from 0.1% by weight to 4% by weight, and more preferably from 0.5% by weight to 3.5% by weight.

Matrix-forming polymers and copolymers.

Suitable matrix-forming polymers and copolymers include polyvinylpyrrolidine (PVP), acrylic polymers and copolymers such as various grades of Eudragit, hydroxypropyl methylcellulose succinate (HPMCAS), as well as other copolymers such as polyethylene glycol-based copolymers such as Soluplus.

Preferred matrix-forming polymers and copolymers are HPMC AS and Soluplus.

The matrix-forming polymers and copolymers may be present in a relative amount that is not particularly limited. Suitable amounts may vary from 0.1 g to 10 g, preferably from 0.2 g to 5 g and more preferably from 0.3 g to 4 g.

Complexing agents.

Suitable complexing agents include cyclodextrins and modified cyclodextrins.

Preferred complexing agents include (2-hydroxypropyl)-P-cyclodextrin and sulfobutyl ether-β-cyclodextrin.

The complexing agents may be present in a relative amount that is not particularly limited. Suitable amounts may range from 0.1 g to 24 g, preferably from 0.1 g to 10 g, and more preferably from 0.1 g to 5 g.

Other types of excipients.

The composition of the present invention may contain additional excipients that are commonly used in the art.

Such additional excipients may include release rate modifiers, a plasticizer, a film-forming agent, a colorant, an antiblocking agent, and/or a pigment for coating the compositions of the present invention. Additional types of excipients that may be present include flavoring agents, sweeteners, antioxidants, absorption accelerators, and/or bulking agents. The relative amount of such excipients is not particularly limited. They can be determined by one skilled in the art based on general knowledge and routine procedures.

Film-forming agents are advantageously used to produce the coherently coated tablets of the invention. Suitable film-forming agents include isomalt, polyvinyl alcohol, polyethylene glycol, maltodextrin, sucrose, xylitol, maltitol, enteric coatings such as materials selected from the group consisting of methyl acrylate-methacrylic acid copolymers, polyvinyl acetate phthalate (PVAP), methyl methacrylate-methacrylic acid copolymers, shellac , sodium alginate and zein. It is preferable to use a combination of film-forming agents comprising polyvinyl alcohol and one or more second agents selected from isomalt, maltodextrin, sucrose, xylitol and maltitol. It is particularly preferred to use a combination of film-forming agents containing at least polyvinyl alcohol and isomalt.

Suitable plasticizers include sorbitol, triacetin, poloxamer, polyethylene glycol, glycerin, propylene glycol, polyethylene glycol monomethyl ether, acetyl tributyl citrate, acetyl triethyl citrate, castor oil, glyceryl monostearate, diacetylated monoglycerides, dibutyl sebacate, diethyl phthalate, triethyl citrate and tributyl citrate.

For each of the above categories of excipients, only one substance or a combination of two or more substances belonging to the same category can be used. Of course, it is not necessary that representatives of every single category be present.

The formulations and compositions of the invention may include the compounds described herein in amorphous particulate form or in any crystalline form. The particle size is not particularly limited. For example, the formulations and compositions may include micronized crystals of the described compounds. Micronization can be carried out on crystals of individual compounds or on mixtures of crystals and parts or whole pharmaceutical excipients or carriers. The average particle size of micronized crystals of the described compound may be, for example, from about 5 to

- 34 046887 about 200 microns or from about 10 to about 110 microns. The compounds of the invention may also be present in the form of a molecular dispersion within a polymer matrix. In yet another embodiment, the compounds of the invention can form complexes with suitable complexing agents, such as cyclodextrins.

The tablet may be made by compression or molding, optionally with one or more additional ingredients. Compressed tablets may be prepared using a binder (eg, gelatin, microcrystalline cellulose, or hydroxypropyl methylcellulose), a lubricant, an inert diluent, a preservative, a disintegrant (eg, sodium starch glycolate or cross-linked sodium carboxymethylcellulose), a surfactant, or a dispersing agent. Molded tablets can be prepared by molding in a suitable machine a mixture of the subject composition moistened with an inert liquid diluent. Tablets and other solid dosage forms, such as dragees, capsules, pills and granules, may optionally be scored or may be formulated with coatings and coatings, such as enteric coatings and other coatings well known in the pharmaceutical formulation art. The excipients described may perform more than one function. For example, fillers or binding agents may also be disintegrating agents, glidants, antiblocking agents, lubricants, sweeteners, and the like.

It should be understood that the composition described may include lyophilized or freeze-dried compounds described herein. For example, compositions are described herein that include the compounds in crystalline and/or amorphous powder forms. Such forms may be reconstituted for use, for example, as an aqueous composition.

Liquid dosage forms for oral administration include pharmaceutically acceptable emulsions, microemulsions, solutions, suspensions, syrups and elixirs. In addition to the subject composition, liquid dosage forms may contain inert diluents commonly used in the art, such as, for example, water or other solvents, solubilizing agents and emulsifiers such as ethyl alcohol, isopropyl alcohol, ethyl carbonate, ethyl acetate, benzyl alcohol , benzyl benzoate, propylene glycol, 1,3-butylene glycol, oils (especially cottonseed, peanut, corn, germ, olive, castor and sesame oils), glycerin, tetrahydrofuryl alcohol, polyethylene glycols and sorbitan fatty acid esters, cyclodextrins and mixtures thereof .

Suspensions, in addition to the subject composition, may contain suspending agents such as, for example, ethoxylated isostearyl alcohols, polyoxyethylene sorbitol and sorbitan esters, microcrystalline cellulose, aluminum metahydroxide, bentonite, agar-agar and tragacanth, and mixtures thereof.

Formulations for rectal or vaginal administration can be presented in the form of suppositories, which can be prepared by mixing the composition with one or more suitable non-irritating excipients or carriers containing, for example, cocoa butter, polyethylene glycol, suppository wax or salicylate, and which are solid when room temperature, but liquid at body temperature and therefore melt in the body cavity and release the active agent. Formulations suitable for vaginal administration also include pessaries, tampons, creams, gels, pastes, foams or sprays containing such carriers as are known in the art to be suitable.

Dosage forms for transdermal administration of the subject composition include powders, sprays, ointments, pastes, creams, lotions, gels, solutions and patches. The active ingredient may be mixed under sterile conditions with a pharmaceutically acceptable carrier and any preservatives, buffers or propellants that may be required.

Ointments, pastes, creams and gels may contain, in addition to the composition in question, excipients such as animal and vegetable fats, oils, waxes, paraffins, starch, tragacanth, cellulose derivatives, polyethylene glycols, silicones, bentonites, silicic acid, talc and zinc oxide or mixtures of them.

Powders and sprays may contain, in addition to the composition in question, excipients such as lactose, talc, silicic acid, aluminum hydroxide, calcium silicates and polyamide powder or mixtures of these substances. Sprays may additionally contain conventional propellants such as chlorofluorocarbons and volatile unsubstituted hydrocarbons such as butane and propane.

The compositions and compounds disclosed herein can alternatively be administered in aerosol form. This is achieved by preparing an aqueous aerosol, liposomal formulation, or solid particles containing the compound. A non-aqueous suspension (eg fluorocarbon propellant) may be used. Sonic nebulizers can be used because they minimize the shearing effect on the agent, which can lead to degradation of the compounds contained in the compositions in question.

Typically, an aqueous aerosol is prepared by forming an aqueous solution or suspension of the composition in question together with conventional pharmaceutically acceptable carriers and stabilizers.

- 35 046887

Carriers and stabilizers vary depending on the requirements of the particular composition under consideration, but typically include non-ionic surfactants (Tweens, pluronics or polyethylene glycol), non-toxic proteins such as serum albumin, sorbitan esters, oleic acid, lecithin, amino acids such as glycine , buffers, salts, sugars or sugar alcohols. Aerosols are usually prepared from isotonic solutions.

It should be noted that the excipients given as examples may have more than one function. For example, fillers or binding agents may also be disintegrating agents, glidants, antiblocking agents, lubricants, sweeteners, and the like.

Pharmaceutical preparations of the present description suitable for parenteral administration contain the subject composition in combination with one or more pharmaceutically acceptable sterile isotonic aqueous or non-aqueous solutions, dispersions, suspensions or emulsions, or sterile powders that can be reconstituted into sterile injectable solutions or dispersions immediately before use, which may contain antioxidants, buffers, bacteriostatics, solutes that render the composition isotonic with the blood of the intended recipient, or suspending agents or thickening agents. For example, provided herein is an aqueous composition that includes the described compound and may further include, for example, dextrose (e.g., about 1 to about 10 weight percent dextrose or about 5 weight percent dextrose in water (D5W).

Examples of suitable aqueous and non-aqueous carriers that may be used in the pharmaceutical compositions herein include water, ethanol, polyols (such as glycerin, propylene glycol, polyethylene glycol and the like) and suitable mixtures thereof, vegetable oils such as olive oil and organic complex Injectable esters such as ethyl oleate and cyclodextrins. Proper fluidity can be maintained, for example, by using coating materials such as lecithin, by maintaining the required particle size in the case of dispersions, and by the use of surfactants.

It should be understood that contemplated formulations, such as oral formulations (eg, a pill or tablet), may be formulated as a controlled release formulation, such as an immediate release formulation, a delayed release formulation, or a combination thereof.

In certain embodiments, the subject compounds may be formulated as a tablet, pill, capsule, or other suitable oral formulation (collectively referred to as tablets). In certain embodiments, the therapeutic dose may be presented in 10 tablets or less. In another example, the therapeutic dose is provided in 50, 40, 30, 20, 15, 10, 5 or 3 tablets.

In a certain embodiment, the described compound is formulated for oral administration in the form of a tablet, capsule, or aqueous solution or suspension. In another embodiment of the tablet form, the tablets are formulated such that the resulting amount of antibacterial agent (or antibacterial agents) presented in 20 tablets, when taken together (e.g., over time) after administration, will provide a therapeutically effective dose and/or a dose that is at least the median effective dose (ED50), for example, a dose at which at least 50% of individuals exhibit a quantitative effect of bacterial cell growth inhibition or protection (eg, a statistically significant reduction in infection). In another embodiment, the tablets may be formulated such that the total amount of antibacterial agent (or antibacterial agents) provided when administered as 10, 5, 2, or 1 tablet would provide a therapeutically effective dose and/or at least an ED50 dose patient (human or non-human mammal). In other embodiments, the amount of antibacterial agent (or antibacterial agents) provided when administered in 20, 10, 5, or 2 tablets taken over a 24-hour period of time will provide a dosage regimen that provides, on average, an average level of antibacterial agent (s) in plasma a therapeutically effective dose and/or at least an ED50 concentration (the concentration for 50% of the maximum effect, for example, inhibition of bacterial cell growth). In other embodiments, the ED50 is less than 100, 10, or 5 times. In other embodiments, a single dose of tablets (1-20 tablets) provides from about 40 mg to 3000 mg of compound(s). These specified unit dosage amounts also apply to other oral dosage forms.

Likewise, the compounds described herein can be formulated for parenteral administration, for example, for subcutaneous, intramuscular or intravenous injection, for example, the antibacterial agent can be presented in a sterile solution or suspension (collectively referred to as injection). In some embodiments, the injection solution may be formulated such that the amount of antibacterial agent (or antibacterial agents) provided, for example, in a bolus injection of from about 0.1 to about 200 ^cc , or dose administered intravenously, provides a dose at least the average effective dose, or less than 100 times the ED50, or less than 10 or 5 times the ED50. Injection

- 36 046887 may be formulated such that the total amount of antibacterial agent (or antibacterial agents) presented (when administered) in injections of 100, 50, 25, 10, 5, 2.5 or 1 cm ³ would provide a therapeutically effective amount and/or the patient's ED50 dose, or less than 100 times the ED50, or less than 10 or 5 times the ED50. In other embodiments, the amount of antibacterial agent (or antibacterial agents) provided upon administration, in a total injection volume of 100 ^cc , 50, 25, 5, or 2 ^cc , at least twice over a 24-hour period of time, provided the dosage regimen providing, on average, an average plasma level of antibacterial agent(s) in a therapeutically effective amount and/or at least a concentration of ED50, or less than 100 times the ED50, or less than 10 or 5 times the ED50. In other embodiments, a single dose injection represents about 40 mg to about 3000 mg, or about 100 mg to about 1000 mg, of antibacterial agent. In the case of v.m. administration, in principle, the same quantity instructions apply. However, the upper limit of the standard dose range may be lower depending on the solubility of the drug compound and the maximum tolerated amount for injection.

Standard dosage forms.

When treating a patient with the pharmaceutical compositions of the present invention by oral administration, a single unit dose of the pharmaceutical composition of the present invention is typically administered once, twice or three times daily. The daily dosage is determined by the doctor in accordance with the instructions given above, taking into account the severity of the infection, gender, body weight, age and general condition of the patient. Preferred oral daily doses are from 40 to 3000 mg, preferably from 100 to 2000 mg. As a result, typical unit dosage forms may range from 40 to 2000 mg, depending on the intended frequency of administration.

In the case of parenteral administration (eg, IV or IM administration), the pharmaceutical compositions of the present invention are typically administered two, three or more times per day. Preferred daily dosages range from 40 to 3000 mg, so that typical unit dosage forms range from 40 to 3000 mg, and preferably from 100 to 1000 mg. The upper limits of the specified ranges depend on their capabilities. For example, in the case of v.m. administration, it may happen that the maximum dose that can be administered at one time is limited due to low solubility and, accordingly, an increase in the volume of drug solution. In such a case, the maximum unit dosage forms are limited to the maximum tolerated dose.

Combinations of drugs.

Also contemplated herein are compositions that include one or more of the disclosed compounds with a second component. The second components in such compositions of the present invention are typically an antibiotic agent other than the described compound. Additional components may also be present, including FabI inhibitors or other antibiotic agents. The contemplated methods of treatment described herein, in some embodiments, may further include the administration of another agent, such as described below. For example, a method of treating a bacterial infection is provided that includes administering the described compound and further includes administering an antibiotic agent or an antibacterial agent described below. The compound described herein and the second component may be part of the same dosage form, or may be formulated as two separate dosage forms. If formulated as two separate dosage forms, the dosage form with the second component can be administered simultaneously, before or after the dosage form with the compound described herein.

Non-limiting examples of antibiotic agents that may be used in the antibacterial compositions herein include cephalosporins, quinolones and fluoroquinolones, penicillins, penicillins and beta-lactamase inhibitors, carbepenems, monobactams, macrolides and lincosamines, glycopeptides, rifampin, oxazolidonones, tetracyclines, streptogramins , sulfonamides and others. Each family contains many members.

Cephalosporins can be further categorized by generation. Non-limiting examples of cephalosporins by generation include the following. Examples of cephalosporins: First generation compounds include Cefadroxil, Cefazolin, Cephalexin, Cefalotin, Cefapirin and Cefradine. Second generation compounds include Cefaclor, Cefamandole, Cefonicid, Cefotetan, Cefoxitin, Cefprozil, Ceftmetazole, Cefuroxime, Cefuroxime Axetil and Loracarbef. The third generation includes Cefdinir, Ceftibuten, Cefditoren, Cefetamet, Cefpodoxime, Cefprozil, Cefuroxime (Axetil), Cefuroxime (sodium), Cefoperazone, Cefixime, Cefotaxime, Cefpodoxime proxetil, Ceftisidime, Ceftizoxime and Ceftriaxone. Fourth generation compounds include Cefepime.

Non-limiting examples of quinolones and fluoroquinolones include Cinoxacin, Ciprofloxacin, Enoxacin, Gatifloxacin, Grepafloxacin, Levofloxacin, Lomefloxacin, Moxifloxacin, Nalidixic acid, Norfloxacin, Ofloxacin, Sparfloxacin, Trovafloxacin, Oxolinic acid, Gemifloxacin and Perfloxacin qin.

Non-limiting examples of penicillins include Amoxicillin, Ampicillin, Bacampicil

- 37 046887 lin, Carbenicillin indanil, Mezlocillin, Piperacillin and Ticarcillin.

Non-limiting examples of penicillin and beta-lactamase inhibitors include Amoxicillin-Clavulanic Acid, Ampicillin-Sulbactam, Benzylpenicillin, Cloxacillin, Dicloxacillin, Methicillin, Oxacillin, Penicillin G (Benzathine, Potassium, Procaine), Penicillin V, Piperacillin+Tazobactam+K, lavulanic acid and Nafcillin. Non-limiting examples of carbapenems include Imipenem-Cilastatin and Meropenem.

A non-limiting example of monobactam includes Aztreonam. Non-limiting examples of macrolides and lincosamines include Azithromycin, Clarithromycin, Clindamycin, Dirithromycin, Erythromycin, Lincomycin and Troleandomycin. Non-limiting examples of glycopeptides include Teicoplanin and Vancomycin. Non-limiting examples of rifampins include Rifabutin, Rifampin and Rifapentine. A non-limiting example of oxazolidonones includes Linezolid. Non-limiting examples of tetracyclines include Demeclocycline, Doxycycline, Metacycline, Minocycline, Oxytetracycline, Tetracycline and Chlortetracycline.

Non-limiting examples of aminoglycosides include Amikacin, Arbakacin, Gentamicin, Kanamycin, Sizomycin, Arbekacin, Neomycin, Netilmicin, Streptomycin, Tobramycin and Paromomycin. A non-limiting example of streptogramins includes Quinopristin+Dalfopristin.

Non-limiting examples of sulfonamides include Mafenide, Silver Sulfadiazine, Sulfacetamide, Sulfadiazine, Sulfamethoxazole, Sulfasalazine, Sulfisoxazole, Trimethoprim Sulfamethoxazole and Sulfamethizole.

Non-limiting examples of other antibiotic agents include Bacitracin, Chloramphenicol, Colistimethate, Fosfomycin, Isoniazid, Methenamine, Metronidazole, Mupirocin, Nitrofurantoin, Nitrofurazone, Novobiocin, Polymyxin B, Spectinomycin, Tobramycin, Tigecycline, Trimethoprim, Colistin, Cycloserine, Capreomycin, amide, para-Aminosalicylic acid and Erythromycin ethylsuccinate + sulfisoxazole.

Therapeutic use.

Medical indications.

The compounds of the present invention can be used to treat bacterial infections in a patient. They are particularly suitable for the treatment of infections caused by N. gonorrhoeae. Other therapeutic indications for which the compounds of the present invention may be used include the following:

Bacillus Spp, in particular Bacillus cereus, Bacillus coagulans, Bacillus megaterium,

Bacillus subtilis, Baclillus anthacis

Bartonella spp.

Brucella Spp, in particular Brucella abortus, Brucella melitensis.

Campylobacter Spp., in particular Campylobacter jejuni, Campylobacter coli.

Enterococcus faecalis, Enterococcus faecium, Legionella pneumophila.

Listeria Spp., in particular Listeria monocytogenes.

Proteus mirabilis, Providencia stuartii.

Rickettsia Spp., in particular Rickettsia rickettsii.

Burkholderia Spp., in particular Burkholderia pseudomallei, Burkholderia mallei, Burkholderia cenocepacia.

Bordetella pertussis, Bordetella parapertussis. Haemophilus influenza, Kingella kingae,

Moraxella catarrhalis.

Streptomyces spp.

Nocardioides spp.

Frankia spp.

Propionibacterium acnes.

Mycobacterium Spp., in particular Mycobacterium smegmatis, Mycobacterium abscessus, Mycobacterium leprae, Mycobacterium tuberculosis, Mycobacterium avium.

Patients.

The compounds of the present invention can be used to treat bacterial infections in patients who are human patients or non-human animals, preferably humans and non-human mammals.

Daily doses.

The dosage of any compound or composition described will vary depending on the symptoms, age and weight of the patient, the nature and severity of the disorder being treated or prevented, the route of administration and the form of the composition in question. Any of the considered

- 38 046887 formulations can be administered as a single dose or divided doses. Dosages of the compositions can be readily determined by methods known to those skilled in the art or as described herein.

In some embodiments, the dosage of the subject compounds is generally in the range of from about 0.01 ng to about 10 g per 1 kg of body weight, in particular in the range from about 1 ng to about 0.1 g per 1 kg, and more particularly , ranging from about 100 ng to about 10 mg per kg.

It may be necessary to determine the effective dose or amount and any possible effects on the time of administration of the composition for any particular composition herein. This may be performed in a routine experiment as described herein, using one or more groups of animals (preferably at least 5 animals per group) or in human trials, if appropriate. The effectiveness of any contemplated composition and method of treatment or prophylaxis can be assessed by administering the composition and assessing the effect of administration by measuring one or more applicable indicators, and comparing the values of these indicators after treatment with the values of the same indicators before treatment.

The exact time of administration and amount of any particular composition under consideration that will provide the most effective treatment in a given patient will depend on the activity, pharmacokinetics and bioavailability of the composition under consideration, the physiological condition of the patient (including age, sex, type and stage of the disease, general physical condition, sensitivity to given dosage and type of drug), route of administration and the like. The instructions provided herein may be used to optimize treatment, such as determining the optimal timing and/or amount of administration, which will require no more than routine experimentation consisting of monitoring the subject and adjusting the dosage and/or timing.

During treatment of a subject, the patient's health can be monitored through the measurement of one or more relevant indicators at predetermined points in time during the treatment period. Treatments, including composition, amounts, timing of administration and composition, can be optimized according to the results of such tracking. The patient can be re-examined periodically to determine the degree of improvement by measuring the same parameters. Based on these re-evaluations, adjustments can be made to the amount of the subject composition administered and possibly the timing of administration.

Treatment may be initiated at lower doses that are less than the optimal dose of the compound. Thereafter, the dose can be increased in small increments until the optimal therapeutic effect is achieved.

Use of subject compositions may reduce the required dosage for any individual agent contained in the compositions since the onset and duration of action of different agents may be complementary to each other.

The toxicity and therapeutic efficacy of the subject compositions can be determined by standard pharmaceutical procedures in cell cultures or experimental animals, for example, to determine LD50 and ED50.

Data obtained from cell culture assays and animal studies can be used to determine dose ranges for use in humans. The dosage of any contemplated composition is preferably within a range of circulating concentrations that include the ED50 with little or no toxicity and/or that produce a statistically significant reduction in infection in at least 50%, for example 60%, 70%, 80%, 90% , 100% of individuals with low or no toxicity, where low toxicity may, for example, mean non-severe and/or predictable and temporary toxicity. For example, the dosage of the composition in question may be selected so that the treatment achieves a reasonable benefit/risk ratio. The dosage may vary within this range depending on the dosage form used and the route of administration used. For the compositions herein, the therapeutically effective dose may be initially estimated from cell culture assays.

Frequency of administration.

The compounds and compositions described herein may be administered one or more times per day, such as once daily (qd), twice daily (bid), three times daily (tid), or four times daily (qd). qid).

Duration of treatment.

The compounds and compositions described herein may be administered for an unlimited period of time. Preferably, they are administered for a period of time sufficient to eradicate the bacterial infection completely or at least to such an extent that the patient's immune system can cope with any remaining pathological bacteria. Typical duration of administration is from 1 day to 2 weeks, and especially from 1 to 5 days. In the case of v.m. administration, the typical duration of administration may range from 1 injection to 4 injections and preferably 1 injection. Multiple injections can be given on the same day, on consecutive days, or on

- 39 046887 intermediate days without introduction.

Methods of treatment.

In another aspect, described herein are methods of treating a bacterial infection, comprising administering to a patient in need thereof a pharmaceutical composition containing the described compound.

In certain embodiments, described herein are methods of treating an N. gonorrhoeae infection in a patient in need thereof, comprising administering a compound of the invention as described herein. Other methods being considered include treating infections caused by one or more of the bacteria or microbes listed above in a patient in need.

For example, described herein is a method of treating N. gonorrhoeae or other bacterial infection in a patient in need thereof, comprising administering a composition containing a disclosed compound, wherein the method further comprises administering, in the same or a separate dosage form, an additional antibacterial or antibiotic agent, as described herein.

Abbreviations.

The following abbreviations are used in this specification

CH Column chromatography

DCM Dichloromethane

N Normal

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G Gram pH Hydrogen ion concentration indicator mole Mole v/v Volume/Volume about. Volume m/z Mass to charge ratio °C degrees Celsius TEA Triethylamine Et ₂ O Diethyl ether HPLC High performance liquid chromatography Vos /77/9С/77-butyl0XYCarb0NIL h hour ml milliliter eq. Equivalent M Weight Me Methyl, CHz MeOH Methanol AcOH Acetic acid THF T ethrahydrofuran DIPEA N,TV-diisopropylethylamine Pd(OAc) ₂ Palladium acetate(P) EtOH Ethanol EDC 1,2-dichloroethane EtOAc Ethyl acetate Et ₃ N Triethylamine water Water CT, CT Room temperature R\tret Hold Time DMF Dimethylformamide ACN Acetonitrile _NH4OAc Ammonium acetate TFC Trifluoroacetic acid GOBT/GOBt 1-G hydroxybenzotriazole those Thin layer chromatography H ₂ O Water

- 41 046887

sat. Saturated rr EDKI Solution 1 -Ethyl-3 -(3-dimethylaminopropyl)carbodiimide NMR Nuclear magnetic resonance With singlet d doublet triplet

m multiplet dd doublet of doublets MHz Megahertz ppm parts per million

N Proton J Combination constant UPLC-MS Ultraperformance liquid chromatography - tandem mass DMSO CDC1 ₃ spectrometry Dimethyl sulf oxide mf Deuterated chloroform SKH Mother liquor zhms Strong cation exchange chromatography GATU Liquid chromatography mass spectrometry HPLC 3-oxide hexafluorophosphate 1-[bis(dimethylamino) methylene]-1R-1,2,3- GBTU triazolo[4,5-0]pyridinium GHTU High performance liquid chromatography TBTU (2-(1H-benzotriazol-1-yl)-1,1,3,3-tetramethyluronium hexafluorophosphate, somi benzotriazoletetramethyluronium hexafluorophosphate tomwi Hexafluorophosphate 2-(6-chloro-1 H-benzotriazol-1 -yl)-1,1,3,3- somvi tetramethylaminium Cy ₂ NCH ₃ RMV O-(benzotriazol-1-yl)-X, Ν, Ν', Ν'-tetramethyluronium tetrafluoroborate Hexafluorophosphate (1-cyano-2-ethoxy-2- TABN oxoethylideneaminooxy)dimethylaminophrolinocarbenium VM or v.m. Hexafluorophosphate N-{[1,3-dimethyl-2,4,6-trioxotetrahydropyrimidine- BB or i.v. 5(6H)-ylideneaminooxy] (dimethylamino)methylene]-M-

methylmethanaminium

4-{[1,3-dimethyl-2,4,6-trioxotetrahydropyrimidine5(6H)ylideneaminooxy](dimethylamino)methylene]morpholin-4-ium hexafluorophosphate

M-Cyclohexyl-M-methylcyclohexanamine p-Methoxybenzyl

Triacetoxyboron hydride sodium

Intramuscular administration

Intravenous administration

The following examples are not intended to limit the scope of the present invention in any way, but are presented only to illustrate the compounds of the invention and their preparation.

Examples

General methods.

All starting materials and solvents were obtained either from commercial sources or obtained according to literature references. Unless otherwise stated, all reactions are stirred. Organic solutions are usually dried over anhydrous magnesium sulfate or sodium sulfate.

Column chromatography is carried out on pre-packed silica cartridges (230-400 mesh, 40-63 µm) using the specified eluent. SCC was purchased from Silicycle and treated with 1 M hydrochloric acid before use. Unless otherwise specified, the cleared re

- 42 046887 the stock mixture is first diluted with MeOH and acidified with a few drops of AcOH. This solution is loaded directly into the SSC and washed with MeOH. The desired product is then eluted by washing with 0.7 M NH3 in MeOH.

Analytical methods.

Analytical LCMS.

Analytical LCMS is carried out using either acidic or basic methods as follows.

Method 1a.

A Waters X-Select CSH C18, 2.5 µm, 4.6 x 30 mm column was eluted with a gradient of 0.1% formic acid in MeCN in 0.1% formic acid in water. The gradient from 5-95% 0.1% formic acid in MeCN occurs between 0.00-3.00 min at 2.5 ml/min with a wash from 3.01-3.5 min at 4.5 ml/min . The column is re-equilibrated to 5% MeCN from 3.60-4.00 min at 2.5 ml/min. The UV spectrum of the eluted peaks is measured using an Agilent 1260 Infinity or Agilent 1200 VWD at 254 nm. Mass spectrum is measured using an Agilent 6120 or Agilent 1956 MSD, run in positive/negative switching mode, or an Agilent 6100 MSD, run in either positive or negative mode.

Method 1b.

A Waters X-Select BEH C18, 2.5 µm, 4.6 x 30 mm column was eluted with a gradient of MeCN in aqueous 10 mM ammonium bicarbonate. The gradient from 5-95% MeCN takes place between 0.00-3.00 min at 2.5 ml/min with a wash from 3.01-3.5 min at 4.5 ml/min. The column is re-equilibrated to 5% MeCN from 3.60-4.00 min at 2.5 ml/min. The UV spectrum of the eluted peaks is measured using an Agilent 1260 Infinity or Agilent 1200 VWD at 254 nm. Mass spectrum is measured using an Agilent 6120 or Agilent 1956 MSD, run in positive/negative switching mode, or an Agilent 6100 MSD, run in either positive or negative mode.

Method 1c.

Product analyzed by UPLC (Acquity and SQD Waters system, water (HPLC grade) UPLC® BEH C18, 50x2.1 mm, 1.7 µm, (0.1% HCOOH in water, 0.1% HCOOH in acetonitrile) 3 min method , 5-95% MeCN/water).

All UPLC-MS analyzes are performed on an Acquity and SQD Waters UPLC system. Masslynx is used to run and analyze experiments using OALogin and automatic or manual integrations. Details of the method are listed below.

Chemical reagents: Water (for HPLC)

Acetonitrile ACN (for HPLC)

Formic acid 98% (LC-MS grade)

Description of the mobile phase.______________________________________________________________

Solution A1: 0.06% HCOOH in water Solution B1: 0.06% HCOOH in acetonitrile Solution A2: Water Solution B2: Acetonitrile Poor flushing Water-ACN (7:3) Strong flushing Water-ACN (1:9) Sealing flush Water-ACN (1:1)

UPLC parameters.

Column: UPLC® VEN S18, 50x2.1 mm, 1.7 µm Mobile phase: A1: 0.06% HCOOH in water B1: 0.06% HCOOH in acetonitrile Gradient Time [min] Flow rate [ml/min] A1 [%] IN 1 [%] 0.00 0.10 3.00 3.10 0.9 0.9 0.9 0.9 95 95 0 95 5 5 100 5 Column temperature: 40°С UV definition: 210-400 nm

- 43 046887

Injection volume: 1-10 µl (as required)

Mass spectroscopy method: carried out based on MS Tune parameters: Routine.ipr, as shown below.

Source AS+ Voltage Capillary (kV) 3.50 Cone (B) thirty Extractor (B) 3 RF Lens (B) OD Temperatures Temp, source (°C) 150 Desolvation rate (°C) 350 Source AS+ Analyzer LM resolution 15.0 NM resolution 15.0 Ion energy 0.6 Increase 1.00

The method is carried out on the basis of information MS Method: pos_neg_3mn_30v. Points per peak: 3846. Total run time: 3.0 min. MS scanning, time 0.00-3.00, mass 100.00-900.00 ER ⁺ .

UPLC-MS mass spectrum was performed on an Acquity Waters SQ sensor (Waters Corporation, Waters Milford, 34 Maple St., Milford, USA) equipped with Empower 2 pro software in positive ionization mode. Ionization conditions: capillary 2.25 kV, cone 160 V, source 100°C, desolvation 150°C; gas flow ^ desolvation 500 l/h, cone 50 l/h.

Analytical UPLC/MS.

Alternatively, analytical UPLC/MS is carried out using either acidic or basic methods, as follows:

Method 2a. A Waters Acquity CSH C18, 1.7 µm, 2.1 x 30 mm column was eluted with a gradient of 0.1% formic acid in MeCN to 0.1% formic acid in water. The gradient is structured with a starting point of 5% MeCN held for 0.0-0.11 min. The 5-95% gradient takes place between 0.11-2.15 min with a washout of 2.15-2.56 min. Re-equilibration of the column to 5% MeCN is carried out from 2.56-2.83 minutes. The UV spectrum of the eluted peaks is measured using an Acquity PDA, and the mass spectrum is recorded using a QDa detector with ESI positive/negative switching.

Method 2b. A Waters Acquity BEH C18, 1.7 µm, 2.1 x 30 mm column was eluted with a gradient of MeCN in aqueous 10 mM ammonium bicarbonate. The gradient is structured with a starting point of 5% held from 0.0-0.11 min. The gradient from 5-95% takes place between 0.11-2.15 min with washing from 2.15-2.56 min. Re-equilibration of the column to 5% MeCN is carried out from 2.56-2.83 minutes. The UV spectrum of the eluted peaks is measured using an Acquity PDA, and the mass spectrum is recorded using a QDa detector with ESI positive/negative switching.

Preparative HPLC.

Preparative HPLC was performed using a Waters Xselect CSH C18, 5 µm, 19x50 mm column using either a gradient of 0.1% formic acid in MeCN in 0.1% aqueous formic acid or a gradient of MeCN in aqueous 10 mM ammonium bicarbonate; or using a Waters Xbridge BEH C18, 5 µm, 19x50 mm column using a MeCN gradient in aqueous 10 mM ammonium bicarbonate. Fractions are collected after determination of UV at a single wavelength measured by a variable wavelength probe on a Gilson 215 preparative HPLC or Varian PrepStar preparative HPLC; using mass and UV at a single wavelength measured on a ZQ single quadrupole mass spectrometer, positive and negative ion electrospray, and a dual wavelength detector on a Waters FractionLynx LCMS.

Preparative chiral high performance liquid chromatography.

Method 3a: Chiralpak® IA column (Daicel Ltd.) (2x25 cm), flow rate 13.5 ml min ^-1 , eluting with (% ethanol) ethanol in 4:1 heptane + 0.2% TFA and chloroform, UV determination at 254 nm. Samples are loaded onto the column through the column unloader pump, pumping chloroform (1.5 ml min ^-1 ) throughout the run, resulting in a combined flow rate of 15 ml min ^-1 .

Method 3b: Chiralpak® IC column (Daicel Ltd.) (2x25 cm), flow rate 13.5 ml min ^-1 , eluting with (% ethanol) ethanol in 4:1 heptane + 0.2% diethylamine, UV detection at 254 nm. Image

- 44 046887 ts are loaded into the column through the unloading pump on the column, pumping chloroform (1.5 ml min ^-1 ) throughout the run, obtaining a combined flow rate of 15 ml min ^-1 .

Analytical chiral high performance liquid chromatography

Method 4a. Chiralpak® IA column (Daicel Ltd.) (4.6 mm x 25 mm), flow rate with a mixture of (% ethanol) ethanol in a 4:1 mixture of heptane + 0.2% TFA and chloroform, UV 254 nm.

Method 4b. Column Chiralpak® IC (Daicel Ltd.) (4.6 mmx25 mm), flow rate ml min ^-1 , elution at ml min ^-1 , eluting with a mixture of (% ethanol) ethanol in 4:1 isohexane + 0.2% diethylamine, UV detection at 254 nm.

¹ H NMR Spectroscopy.

The 1H NMR spectrum was obtained on a Bruker Avance III spectrometer at 400 MGz using residual non-deuterated solvent as a reference.

Example 1. Synthesis of (E)-N-((4-amino-3-methylbenzofuran-2-yl)methyl)-N-methyl-3-(8-oxo-6,7,8,9tetrahydro-5H-pyrido[ 2,3-b]azepin-3-yl)acrylamide (compound 5).

General synthesis scheme.

Reaction conditions: a) MeNH ₂ , EtOH; b) BH ₃ , THF, reflux; c) GATU, DIPEA DMF.

The synthesis of (E)-3-(8-oxo-6,7,8,9-tetrahydro-5H-pyrido[2,3-b]azepin-3-yl)acrylic acid (compound 4) was previously described in AFFINIUM PHARMECEUTICALS, INC: WO2007/67416, 2007, A2.

Step 1. 4-Amino-H3-dimethylbenzofuran-2-carboxamide (compound 2).

Ethyl 4-amino-3-methylbenzofuran-2-carboxylate 1 (0.1 g, 0.46 mmol) was dissolved in methamine (10 ml, 80 mmol, 33% in EtOH), and the reaction mixture was stirred at RT for ~36 h. The solvent was removed in vacuo to obtain the desired product 2 as a pale brown crystalline solid (0.097 g, quantitative). R _t 1.25 min (method 1a); m/z 205 [M+H] ⁺ (ER ⁺ ), ¹ H NMR (400 MHz, DMSO-06): δ, ppm 8.24 (q, J=4.6 Hz, 1H), 7, 07 (t, J=8.0 Hz, 1H), 6.68 (dd, J=8.2, 0.8 Hz, 1H), 6.43 (dd, J=7.9, 0.8 Hz , 1H), 5.41 (s, 2H), 2.74 (d, J=4.7 Hz, 3H), 2.72 (s, 3H).

Step 2. 3-Methyl-2-((methylamino)methyl)benzofuran-4-amine (compound 3).

To a suspension of compound 2 (300 mg, 1.47 mmol) in dry THF (30 ml) at 0°C add boron tetrahydrofuran complex (7.34 ml, 7.34 mmol, 1 M in THF). After warming to RT, the reaction mixture is refluxed for 3 hours. The mixture is quenched with methanol (20 ml) while still refluxing, then 1 N hydrochloric acid (2 ml) is added and the mixture is continued to reflux within 30 min. The mixture is cooled to RT, then evaporated to dryness. The residue is taken up in methanol (50 ml) and applied to an SCC column. The column is washed with methanol (50 ml) and the product is eluted with 10% methanolic ammonia. The solvent was removed in vacuo to give the desired product 3 as a yellow oil (0.20 g, 68%). Rt 0.17 min (method 1a); m/z 160 [M - NHCH3] ⁺ (ER+). 1H NMR (400 MHz, DMSO-06): δ, ppm 6.88 (t, J=7.9 Hz, 1H), 6.63 (dd, J=8.1 Hz, 0.8 Hz, 1H), 6.36 (dd, J=7.8 Hz, 0.8 Hz, 1H), 5.10 (s, 2H), 3.66 (s, 2H), 2.34 (s, 3H) , 2.24 (s, 3H).

GATU, DIPEA

DMF,

- 45 046887

Step 3. (E)-U-((4-Amino-3-methylbenzofuran-2-yl)methyl)-U-methyl-3-(8-ox.o-6,7,8,9-tetrahydro5H-pyrido [2,3-b]azepin-3-yl)acrylamide (compound 5).

Solution of 3-methyl-2-((methylamino)methyl)benzofuran-4-amine 3 (0.07 g, 0.35 mmol), (E)-3-(8-oxo6,7,8,9-tetrahydro- 5H-pyrido[2,3-b]azepin-3-yl)acrylic acid 4 (0.07 g, 0.29 mmol) and DIPEA (0.26 ml, 1.47 mmol) in DMF (2 ml) stirred within 10 min. GATU (0.17 g, 0.44 mmol) was then added in one portion and the reaction mixture was stirred for 2 hours. Water (20 ml) was added to the mixture and the resulting precipitate was collected by filtration. The solid is washed with water (10 ml) and dried on filter paper, then dissolved in as little DMSO as possible and precipitated by adding water. The solid was collected, then the crude product was further purified by chromatography (0-10% MeOH in DCM) to give compound 5 as a tan solid (0.04 g, 29%). Rt 1.54 min (method 1a); m/z 405 [M+H]+ (ER ⁺ ), ^1H NMR (400 MHz, DMSO^ ₆ , 363 K): δ, ppm 9.56 (s, 1H), 8.48 (d, J=2.2 Hz, 1H), 8.02 (d, J=2.2 Hz, 1H), 7.51 (d, J=15.4 Hz, 1H), 7.27 (d, J= 15.6 Hz, 1H), 6.92 (t, J=8.0 Hz, 1H), 6.66 (dd, J=8.2 Hz, 0.9 Hz, 1H), 6.41 (dd , J=7.8 Hz, 0.8 Hz, 1H), 4.93 (s, 2H), 4.77 (s, 2H), 3.07 (s, 3H), 2.77 (t, J =7.1 Hz, 2H), 2.43 (s, 3H), 2.32 (t, J=7.2 Hz, 2H), 2.15 (p, J=7.7, 7.3 Hz , 2H).

Example 2. Synthesis of (E)-N-((4-amino-3-methylbenzofuran-2-yl)methyl)-N-methyl-3-(4-oxo-2,3,4,5tetrahydro-1H-pyrido[ 2,3-b][1,4]diazepin-8-yl)acrylamide (compound 7).

General synthesis scheme.

TFA N O ⁶ 7

Reaction conditions: (a) GATU, DIPEA, DMF.

The synthesis of (E)-3-(4-oxo-2,3,4,5-tetrahydro-1H-pyrido[2,3-b][1,4]diazepin-8yl)acrylic acid hydrochloride (compound 6) was previously described in AFFINIUM PHARMECEUTICALS, INC: WO2007/67416, 2007, A2; AURIGENE DISCOVERY TECHNOLOGIES LIMITED; AURIGENE DISCOVERY TECHNOLOGIES LIMITED: WO2013/80222, 2013, A1; Ramnauth Jailall et al., Bioorg. Med. Chem. Lett, 2009, 19, pp. 5359-5362.

6 7

Step 1. (E)-N-((4-Amino-3-methylbenzofuran-2-yl)methyl)-N-methyl-3-(4-oxo-2,3,4,5-tetrahydro1H-pyrido[2 ,3-b][1,4]diazepin-8-yl)acrylamide (compound 7).

Solution of 3-methyl-2-((methylamino)methyl)benzofuran-4-amine 3 (0.05 g, 0.26 mmol), TFA salt (E)-3(4-oxo-2,3,4,5 -tetrahydro-III-pyrido[2,3-b][1,4]diazepin-8-yl)acrylic acid 6 (0.05 g, 0.22 mmol) and DIPEA (0.38 ml, 2.19 mmol ) in DMF (2 ml) and stir for 10 min. 2-(3H-[1,2,3]triazolo[4,5-b]pyridin-3-yl)-1,1,3,3-tetramethylisouronium hexafluorophosphate (0.13 g, 0.33 mmol) was then added one portion, and the reaction mixture was stirred for 2 hours. Water (20 ml) was added to the mixture, and the resulting precipitate was collected by filtration. The solid is washed with water (10 ml) and dried on filter paper, then dissolved in as little DMSO as possible and precipitated by adding water. The solid was collected, then the crude product was further purified by chromatography (0-10% MeOH in DCM) to give the desired product 7 as a yellow solid (0.05 g, 53%). R _t 1.43 min (method 1a); m/z 406 [M+H]+ (ER+). ¹ H NMR (400 MHz, DMSO-de, 363 K): δ, ppm 9.14 (s, 1H), 7.97 (d, J=1.9 Hz, 1H), 7.45-7 .34 (m, 2H), 7.09 (d, J=15.7 Hz, 1H), 6.92 (t, J=8.0 Hz, 1H), 6.66 (dd, J=8, 2 Hz, 0.8 Hz, 1H), 6.41 (dd, J=7.8 Hz, 0.8 Hz, 1H), 5.82 (d, J=4.6 Hz, 1H), 4, 93 (s, 2H), 4.74 (s, 2H), 3.48-3.40 (m, 2H), 3.05 (s, 3H), 2.69-2.62 (m, 2H) , 2.43 (s, 3H).

- 46 046887

Example 3. Synthesis of (H)-\-methyl-\-((3-methylbenzofur;1n-2-yl)methyl)-3-(3-oxo-1,2,3,4tetrahydropyrido[2,3-b] pyrazin-7-yl)acrylamide (compound 10).

General synthesis scheme.

A

10

Reaction conditions: a) Pd-162, Bu4N ⁺ Cl ^- , Cy ₂ NCH ₃ dioxane 80 g. C, 2 hours

The synthesis of 7-bromo-1,4-dihydropyrido[2,3-b]pyrazin-3(2H)-one (compound 8) was previously described in CEPHALON, INC - WO2007/130468, 2007, A2.

NMethyl-N-((3-methylbenzofuran-2-yl)methyl)acrylamide (compound 9) was prepared as described in AFFINIUM PHARMACEUTICALS, INC. patents. - WO2007/67416, 2007, A2 and VITAS PHARMA RESEARCH PRIVATE LIMITED, WO2013/42035, 2013, A1.

Step 1. (E)-N-Methyl-N-((3-methylbenzofuran-2-yl)methyl)-3-(3-oxo-1,2,3,4-tetrahydropyrido[2,3EDpyrazin^-yl^ krylamide (compound 10).

Namethyl-N-((3-methylbenzofuran-2-yl)methyl)acrylamide 9 (23.12 mg, 0.101 mmol), 7-bromo-1,2-dihydropyrido[2,3-b]pyrazine is loaded into the reaction tube -3(4H)-one 8 (23 mg, 0.10 mmol), tetrabutylammonium chloride hydrate (2.98 mg, 10.09 µmol), [P(tBu) ₃ ]Pd(crotyl)Cl (Pd-162) (4.03 mg, 10.09 µmol). The tube is then rinsed with nitrogen (5 min). 1,4-dioxane (3 ml) and N-cyclohexyl-N-methylcyclohexanamine (43.2 µl, 0.20 mmol) were added and the reaction mixture was purged with nitrogen (5 min). The mixture is heated to 80°C for 2 hours and cooled to room temperature. The solvent is evaporated to dryness. The residue is taken up in DCM (5 ml) and sat. NaHCO ₃ (5 ml). The aqueous phase is separated and extracted with DCM (2x5 ml). The combined organic phases are passed through a phase separator and concentrated in vacuo. The crude product was purified by silica chromatography (0-10% MeOH/DCM) to give the title compound 10 as a yellow solid (6.2 mg, 0.016 mmol, 16% yield). Rt 1.20 min (method 2a); 377.0 [M+H] ⁺ (ER ⁺ ), 1H

NMR Obtained at 360K (400 MHz, DMSO-06): δ, ppm 10.49 (s, 1H), 7.83 (d, J=1.9 Hz, 1H), 7.58-7.54 (m, 1H), 7.47 (dt, J=8.2, 0.8 Hz, 1H), 7.42 (d, J=15.4 Hz, 1H), 7.31-7.23 ( m, 3H), 7.07 (d, J=15.9 Hz, 1H), 5.99 (s, 1H), 4.84 (s, 2H), 3.85 (d, J=1.9 Hz, 2H), 3.10 (s, 3H), 2.28 (s, 3H).

Example 4. Synthesis of (E)-3-(2,2-dimethyl-3-oxo-1,2,3,4-tetrahydropyrido[2,3-b]pyrazin-7-yl)-Nmethyl-N-(( 3-methylbenzofuran-2-yl)methyl)acrylamide (compound 19).

General synthesis scheme.

Reaction conditions: a) 2-amino-2-methylpropanoic acid 12, K2CO3/THF reflux, overnight; b) Fe, water, EtOH, NH ₄ Cl; c) tri-(o-tolyl)phosphine, Pd(AcO) ₂ , DIPEA, DMF:propionitrile (1:4 v/v); d) TPA/DXM; e) T ₃ P, DIPEA, DMF.

- 47 046887

Step 1. 2-((5-Bromo-2-nitropyridin-3-yl)amino)-2-methylpropanoic acid (compound 13).

A mixture of 5-bromo-3-fluoro-2-nitropyridine 11 (30.0 g, 135.7 mmol), K ₂ CO ₃ (38.96 g, 282.31 mmol) and 2-amino-2-methylpropanoic acid 12 (20 .9 g, 203.6 mmol) in THF (500 ml) was heated at 90°C for 16 hours. The progress of the reaction was monitored by TLC. The reaction mixture is cooled to RT and concentrated to dryness. The crude product is dissolved in water (500 ml) and washed with ethyl acetate (100 ml). The aqueous layer is acidified with 1N HCl solution to pH 2 and extracted with ethyl acetate (2x250 ml). The organic layer is dried over NaSO4, filtered and concentrated in vacuo to give the crude compound. Add 50 mL of diethyl ether to the crude compound and stir for 30 min, filter the solid and dry under vacuum to give the title compound 13 as a yellow solid (26.0 g, 63% yield), m/z 304/ 306 [M+H]+ (ER ⁺ ), ¹ H NMR (400 MHz, DMSO^): δ, ppm 13.48 (s, 1H), 7.90 (s, 1H), 8.10 ( s, 1H), 7.97 (s, 1H), 7.39 (s, 1H), 1.60 (s, 6H).

Step 2. 7-Bromo-2,2-dimethyl-1,4-dihydropyrido[2,3-b]pyrazin-3(2H)-one (compound 14).

To a stirred solution of 2-((5-bromo-2-nitropyridin-3-yl)amino)-2-methylpropanoic acid 13 (26.0 g, 85.4 mmol) in EtOH:water (1:1, 520 ml) add NH ₄ Cl (10.42 g, 194 mmol), then iron powder (10.31 g, 184.6 mmol). The reaction mixture is stirred at 80°C for 2 hours. The progress of the reaction is monitored by TLC. The reaction mixture is cooled to RT and diluted with ethyl acetate (500 ml), filtered through Celite® and washed with THF (200 ml), MeOH (200 ml). The filtrate is concentrated in vacuo to obtain the crude compound. Add 50 mL of diethyl ether to the crude compound and stir for 30 min, filter the solid and dry under vacuum to give the title compound 14 as an off-white solid (12.5 g, 57% yield), m/z 256/ 258 [M+H] ⁺ (ER ⁺ ), 1H NMR (400 MHz, DMSO-ph): δ, ppm 10.83 (s, 1H), 7.63 (d, J=2.0 Hz , 1H), 7.09 (d, J=2.0 Hz, 1H), 6.54 (s, 1H), 1.25 (s, 6H).

Step 3. tert-Butyl (E)-3-(2,2-dimethyl-3-oxo-1,2,3,4-tetrahydropyrudo[2,3-b]nyrazin-7yl)acrylate (compound 16).

In a hermetically sealed test tube, stir a solution of 7-bromo-2,2-dimethyl-1,4dihydropyrido[2,3-b]pyrazin-3(2H)-one 14 (5.0 g, 19.5 mmol) and DIPEA ( 14.3 g, 78.12 mmol) in DMF:propionitrile (1:4, 75 ml) and purged with nitrogen gas for 10 min. Then add tert-butyl acrylate 15 (7.5 g, 58.5 mmol), tri-(o-tolyl)phosphine (1.18 g, 3.9 mmol) and palladium(P) acetate (4.37 g, 1 .95 mmol) to the reaction mixture and again purged with nitrogen gas for 5 minutes. The reaction mixture is heated at 90°C for 18 hours. The progress of the reaction is monitored by TLC and LCMS. The reaction mixture is cooled to RT and diluted with 10% MeOH in DCM (50 ml), filtered through Celite®, washed with 10% MeOH in DCM (50 ml) and the filtrate is concentrated in vacuo to give the crude product, which is triturated with diethyl ether (20 ml ) and n-pentane (20 ml), filter and dry under vacuum to obtain the title compound 16. Note: The reaction is carried out in another batch (each 5 g, 10.0 g total) to obtain a total of 6.5 g, 55% as yellow solid, m/z 304.41 [M+H] ⁺ (ER ⁺ ), 1H NMR (400 MHz, DMSO-de): δ, ppm, 10.88 (s, 1H) , 7.87 (s, 1H), 7.457.41 (d, J=16.0 Hz, 1H), 7.18 (s, 1H), 6.32-6.27 (m, 2H), 1, 47 (s, 9H), 1.26 (s, 6H).

17

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Step 4. ((E)-3-(2,2-Dimethyl-3-oxo-1,2,3,4-tetrahydropyrido[2,3-b]pyrazin-7yl)acrylic acid trifluoroacetate (compound 17).

Trifluoroacetic acid (30.0 ml) was added to the cold suspension of tert-butyl (E)-3-(2,2dimethyl-3-oxo-1,2,3,4-tetrahydropyrido[2,3-b]pyrazin-7- yl)acrylate 16 (6.0 g, 19.8 mmol) in DCM (60 ml) at 0°C. The reaction mixture is stirred at RT for 3 hours. The progress of the reaction is monitored by TLC and LCMS, then concentrated in vacuo. The residue was triturated with diethyl ether (2x50 ml) and the solid was collected by filtration to give the title compound 17 as a yellow solid (4.5 g, 92% yield), m/z 248.05 [M+H]+ ( ER ⁺ ). ^1H NMR (400 MHz, DMSO- ₁₆ ): δ, ppm 12.33 (s, 1H), 10.89 (s, 1H), 9.78 (s, 1H), 7.86 (d , J=1.6 Hz, 1H), 7.50-7.46 (d, J=16.0 Hz, 1H), 7.19 (d, J=1.2 Hz, 1H), 6.36 (s, 1H), 6.32-6.28 (d, J=16.0 Hz, 1H), 1.26 (s, 6H).

The synthesis of Nmethyl-1-(3-methylbenzofuran-2-yl)methanamine (compound 18) was previously described by AFFINIUM PHARMACEUTICALS, INC. - WO2008/98374, 2008, A1 and/or VITAS PHARMA RESEARCH PRIVATE LIMITED, WO2013/42035, 2013, A1.

Stage 5.

(E)-3-(2,2-Dimethyl-3-oxo-1,2,3,4-tetrahydropyrido[2,3-b]pyrazin-7-yl)-X-methyl-X-((3methylbenzofuran- 2-yl)methyl)acrylamide (compound 19).

DIPEA (16.7.0 mL, 91.0 mmol) was added dropwise to a stirred solution of U-methyl-1-(3methylbenzofuran-2-yl)methanamine 18 (3.8 g, 18.21 mmol) and trifluoroacetate (E )-3-(2,2-dimethyl-3-oxo-DD-tetrahydropyridoRD-Cpyrazin-yl-acrylic acid 17 (4.5 g, 13.04 mmol) in DMF (25 ml) and THF (25 ml) The reaction mixture was stirred for 5 minutes, then T ₃ P 50% in ethyl acetate (34.7 ml, 54.6 mmol) was added at 0°C, and the reaction mixture was stirred for another 4 hours at RT. The progress of the reaction was monitored by TLC. The reaction mixture is diluted with water (500 ml) and the solid is collected, washed with water (2x50 ml) and dried under vacuum. The crude compound is taken into diethyl ether (50 ml) and stirred for 15 min at RT. The solid is filtered and washed with diethyl. ether (25 ml), dry under vacuum to give (E)-3-(2,2-dimethyl-3-oxo-1,2,3,4tetrahydropyrido[2,3-b]pyrazin-7-yl)-N -methyl-N-((3-methylbenzofuran-2-yl)methyl)acrylamide 19 as a light yellow solid (3.55 g, 67.5% yield). The filtrate was concentrated to obtain 3.5 g of mixture, m/z 405.21 [M+H] ⁺ (ER ⁺ ), ¹ H NMR (400 MHz, DMSO^ ₆ ): δ, ppm 10.84 (s, 1H), 7.90 (s, 1H), 7.57-7.55 (d, J=7.2 Hz, 1H), 7.50-7.42 (m, 2H), 7.30-7 .22 (m, 3H), 7.03-6.99 (d, J=15.2 Hz, 1H), 6.35-6.33 (d, J=9.6, Hz, 1H), 4 .94-4.78 (d, J=64.8 Hz, 2H), 3.16-2.94 (d, 3H), 2.26 (s, 3H), 1.25 (s, 6H).

Example 5. Synthesis of (e)-(8-(3-(methyl((3-methylbenzofuran-2-yl)methyl)amino)-3-oxoprop-1-en-1-yl)-4oxo-1,2, 3,4-tetrahydro-5h-pyrido[2,3-b][1,4]diazepin-5-yl)methylphosphate 2-hydroxyethane-1-aminium (compound 23).

General synthesis scheme.

Reaction conditions: a) 1) 18-crown-6, NMP/N-ethylpyrrolidone, RT; 2) t-BuOK at -20°C to -35°C, then chloromethyl bis-[2-(trimethylsilyl)ethyl]phosphate 21; b) TPA/DCM, long-term processing.

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Total synthesis of (E)-N-methyl-N-((3-methylbenzofuran-2-yl)methyl)-3-(4-oxo-2,3,4,5-tetrahydro-Spirido[2,3-b] [1,4]diazepin-8-yl)acrylamide (compound 20) is described previously in Bioorganic & Medicinal Chemistry Letters (2009), 19(18), 5359-5362 and WO 2007067416.

Step 1. bis-(2-(Trimethylsilyl)ethyl)phosphate (E)-(8-(3-(methyl((3-methylbenzofurαn-2-yl)methyl)amino)-3-oxoprop-1-en-1 -yl)-4-oxo-1,2,3,4-tetrahydro-5H-pyrido[2,3-b][1,4]diazepin-5-yl)methyl (compound 22).

In a hermetically sealed 10 ml test tube under nitrogen containing (Ε)-Ν-μο™λ-Ν-((3methylbenzofuran-2-yl)methyl)-3-(4-oxo-2,3,4,5-tetrahydro- 1H-pyrido[2,3-b][1,4]diazepin-8-yl)acrylamide 20 (250 mg; 1 eq.; previously ground with dry stirring for 10 min) and 18-crown-6 (208 mg ; 1.24 eq.), add N-methyl-2-pyrrolidone (1.0 ml; 4 vol.) and N-ethylpyrrolidone (0.275 ml; 1.10 vol.). The mixture is then stirred at 25°C for 10 min and degassing cycles (5 cycles; vacuum/nitrogen) are carried out. The reaction mixture is then cooled to -20°C (not measured; bath temp -30°C) and potassium tert-butoxide (82 mg; 1.1 eq.) is added in portions over 2 minutes. The reaction mixture is then subjected to degassing cycles again (3 cycles; vacuum/nitrogen) and then stirred at -20°C (bath temperature: -30°C) for 1 hour 30 minutes until almost completely solubilized (dark green mixture). The reaction mixture is cooled to -35°C (not measured; bath temp -45°C) and degassed (5 cycles; vacuum/nitrogen) with chloromethyl bis-[2-(trimethylsilyl)ethyl]phosphate 21 (Carbogen Amcis) solution ( 1.058 g; 1.4 eq; 29.1 w/w% in heptane) added over 4 min while maintaining temperature at -35°C (not measured; temp, bath -45°C). The reaction mixture (two-phase) is stirred at -35°C for 16 hours until almost complete, as shown by UPLC/MS: (starting material: R _t = 1.62 min; 34.5% (AUC); ER ⁺ : 391 .5. Product: Rt=2.50 min; 34.25% (ER ⁺ : 701.79). Na Methyl-2-pyrrolidone (0.64 ml), methylcyclohexane (1.25 ml), toluene (0.35 ml) and water (0.37 ml) were added to the reaction mixture, maintaining the temperature at -20°C (not measured ; bath temperature -30°C). Then, after stirring and phase separation, the lower phase is extracted with MTBE (3 x 2 ml), maintaining a temperature of -20°C. All upper phases are then combined and washed with semi-saturated sodium chloride solution (2 x 6 ml). All washings are carried out while maintaining a temperature of -10°C (not measured; bath temperature -20°C). The organic phase is then dried over sodium sulfate, filtered and washed with MTBE (2x3 ml) at 25°C. Concentrate to dryness, keeping the bath temperature below 25°C to obtain the crude compound as an orange foam (m=150.73 mg; purity: 88.36% (AUC)). Store overnight under nitrogen at -25°C. The crude compound is purified by normal phase chromatography (SiO2; 40-63 µm). Conditioning is carried out with n-heptane, then the crude compound is solubilized in a mixture of n-heptane/MTBE/DIPEA: 2/1/0.75% (3 ml) and MTBE (2 ml) and injected. Elution is carried out with cold eluents (stored in an ice maker for several hours). First eluent (n-heptane/MTBE/DIPEA: 2/1/0.75%), second eluent (MTBE/DIPEA: 100/0.2%), third eluent (methyl acetate/DIPEA: 100/0.2%) . The product is finally eluted with a third eluent. Fractions were combined and concentrated to dryness (bath temperature: 25°C) to obtain the title compound 22 as a yellow oil (m=122.5 mg; purity: 96% (AUC); ER ⁺ : 701.7; yield: 26 ,5%). Store at -25°C until next step.

Step 2. (E)-(8-(3-(Methyl((3-methylbenzofuran-2-yl)methyl)amino)-3-oxoprop-1-en-1-yl)-4-oxo1,2,3 ,4-tetrahydro-5H-pyrido[2,3-b][1,4]diazepin-5-yl)methylphosphate 2-hydroxyethane-1-aminium (compound 23).

In a hermetically sealed 2.5 ml test tube under nitrogen, prepare a solution of TPA (0.369 ml; 19.3 eq.) in anhydrous dichloromethane (0.32 ml; 2.6 volumes) at -20°C, this solution is called No. 1. B hermetically sealed

- 50 046887 In a 2.5 ml test tube under nitrogen, prepare a solution of bis-(2-(trimethylsilyl)ethyl)phosphate (E)-(8-(3(methyl((3-methylbenzofuran-2-yl)methyl)amino)- 3-oxoprop-1-en-1-yl)-4-oxo-1,2,3,4-tetrahydro-5H-pyrido[2,3-b][1,4]diazepin-5-yl)methyl 22 (123 mg; 1 eq.) in anhydrous toluene (0.408 ml; 3.33 volume) at -10°C, this solution is called solution No. 2. Then solution No. 2 is added to solution No. 1 with stirring for 10 minutes, keeping temperature -20°C (not measured; bath temperature -30°C). the tube containing solution No. 2 is then washed with anhydrous dichloromethane (0.404 ml; 3.3 volumes), which is then added to solution No. 1. The reaction mixture is then stirred for 25 min under nitrogen while maintaining the temperature at -20°C (not measured; the temperature is adjusted while keeping the yellowish reaction mixture at the freezing point (FLC): lightly shaking). Completion of hydrolysis is confirmed by UPLC/MS (product: Rt=1.34 min; ER ^- : 499.3. Starting material: Rt=2.49 min; 0% (IIC)). In a hermetically sealed 20 ml test tube under nitrogen, prepare a solution of ethanolamine (274 mg; 26.5 eq.) in a mixture of acetone (1.814 ml; 14.8 volumes) and water (0.050 ml; 0.41 volumes) at 0°C (not measured; bath temperature: -5°C), this solution is called solution No. 3. The previous reaction mixture is then cooled to -20°C (not measured; slightly above freezing point) and added via cannula to solution No. 3 with gentle stirring (100). rpm; can be increased to obtain a homogeneous solution (two-phase) for 10 minutes, maintaining the temperature at 0°C (bath temperature -5°C). The tube containing the reaction mixture was then washed with dichloromethane (0.208 ml; 1.7 volumes) then added to solution No. 3. The final reaction mixture was then stirred at 200 rpm. at 0°C (measured internally) for 30 minutes and store at 4°C for 14 hours in the refrigerator until sedimentation is observed. The reaction mixture is heated up to 25°C and filtered through a glass filter. The resulting cake is washed with ethanol (2x2 ml), but complete dissolution is observed. The filtrate is concentrated to dryness (bath temperature 25°C), and the resulting oil is dissolved in a mixture of acetone (1.5 ml), toluene (1.5 ml) and MTBE (3 ml). The resulting solution is stored at -25°C for 1 hour 30 minutes until the sediment completely settles out. The suspension is filtered through a glass filter and the resulting solid is washed with a mixture of 1-butanol (2 ml) and MTBE (4 ml), then MTBE (2x2 ml) and finally 2-butanone (1x3 ml and 1x2 ml). However, after drying, NMR analysis on the isolated solid is washed only with TPA-ethanolamine salt. The filtrate is stored at -25°C for 16 hours until precipitation is observed. The suspension is filtered through a glass filter and the resulting yellow solid is washed several times with 2-butanone (2 ml) until an acceptable amount of TFA salt of ethanolamine is measured by NMR. Finally, after drying at 25°C under vacuum, the title compound 23 was obtained as a yellow powder (m=42.41 mg; purity: 100% (IIC); ER ^- : 499.3; 2 equiv. ethanolamine via NMR ; 2.4 wt.% TFA salt of ethanolamine; yield: 40.6%). 1H NMR (400 MHz, D2O): δ, ppm 8.13 (d, J=1.7 Hz, 1H), 7.47 (dd, J=1.8 Hz, 6.6 Hz, 1H) , 7.43-7.08 (m, 5H), 6.87 (d, J=15.8 Hz, 1H), 5.46 (d, J=5.6 Hz, 2H), 4.68 ( s, 1H), 4.60 (s, 1H), 3.72 (t, J=5.2 Hz, 4H), 3.65-3.55 (m, 2H), 3.05 (t, J =5.2 Hz, 4H), 3.05 (s, 1.8H), 2.92 (s, 1.2H), 2.53-2.42 (m, 2H, m), 2.09 ( s, 1.8H), 2.08 (s, 1.2H) (rotamers).

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Example 6. Synthesis of (E)-3-(3-(hydroxymethyl)-3-methyl-4-oxo-2,3,4,5-tetrahydro-1H-pyrido[2,3b][1,4]diazepine- 8-yl)-N-methyl-N-((3-methylbenzofuran-2-yl)methyl)acrylamide (compound 31), as well as two enantiomers (S,E)-3-(3-(hydroxymethyl)-3- methyl-4-oxo-2,3,4,5-tetrahydro-1H-pyrido[2,3b][1,4]diazepin-8-yl)-N-methyl-N-((3-methylbenzofuran-2- yl)methyl)acrylamide (compound 32) and (R,E)3-(3-(hydroxymethyl)-3-methyl-4-oxo-2,3,4,5-tetrahydro-1H-pyrido[2,3- b][1,4]diazepin-8-yl)-N-methylN((3-methylbenzofuran-2-yl)methyl)acrylamide (compound 33).

General synthesis scheme.

Reaction conditions: a) K ₂ CO ₃ , (HCHO) _n , EtOH; b) HCl in 1,4 dioxane, PtO ₂ -H ₂ O, EtOH; c) 5-bromo-3fluoro-2-nitropyridine 11, TEA, EtOH, A; d) LiOH, water/THF; e) Fe, NH4Cl, EtOH, H ₂ O, 90°C; f) GATU, DIPEA, DMF; g) Pd-162, Cy ₂ NMe, NBu4Cl, 1,4-dioxane, 80°C; h) chiral separation.

Step 1. Ethyl 2-cyano-3-hydroxy-2-methylpropanoate (compound 25).

A mixture of paraformaldehyde (3.54 g, 118 mmol), ethyl 2-cyanopropanoate 24 (TCI - Tokyo Chemical Industry) (10 g, 79 mmol) and potassium carbonate (32.6 g, 236 mmol) in ethanol (500 ml) is stirred for 18 hours. The solid is removed by filtration through a pad of Celite® and the filtrate is evaporated to dryness. The crude product was purified by silica chromatography (0-50% EtOAc/isohexane) to give the title compound 25 as a clear, colorless oil (6.01 g, 48%), ¹ H NMR (400 MHz, CDCl ₃ ): δ , ppm 4.30 (kv, J=7.1 Hz, 2H), 3.98-3.83 (m, 2H), 2.59 (s, 1H), 1.58 (s, 3H) , 1.34 (t, J=7.1 Hz, 3H).

EtO ₂ C CN PtO ₂ *H ₂ O, EtOH EtO ₂ C /

HO AC but, JX . NHo HCl

HCl in 1,4-dioxane ¹

26

Step 2. Ethyl 3-amino-2-(hydroxymethyl)-2-methylpropanoate hydrochloride (compound 26).

A mixture of platinum oxide (CO) (0.14 g, 0.64 mmol), ethyl 2-cyano-3-hydroxy-2-methylpropanoate 25 (1.00 g, 6.36 mmol) and HCl (4 M in 1.4 β-dioxane, 3.18 ml, 12.73 mmol) in ethanol solvent (12 ml) was stirred under a hydrogen atmosphere at 5 bar for 18 hours. The catalyst was removed by filtration and the filtrate was evaporated to dryness. The residue was dissolved in acetonitrile (10 ml) and evaporated to dryness to give the title compound 26 as a colorless gum (1.38 g, quant. yield), 1H NMR (400 MHz, DMSO^) δ 8.11 (s, 3H ), 5.31 (s, 1H), 4.13-4.07 (m, 2H), 3.64-3.44 (m, 2H), 3.09-2.91 (m, 2H), 1.21 (t, J=7.1 Hz, 3H), 1.13 (s, 3H).

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Step 3. Ethyl 3-((5-bromo-2-nitropyridin-3-yl)amino)-2-(hydroxymethyl)-2-methylpropanoate (compound 27).

Ethyl 3-amino-2-(hydroxymethyl)-2-methylpropanoate hydrochloride mixture 26 (322 mg,

1.63 mmol), triethylamine (757 μl, 5.43 mmol) and 5-bromo-3-fluoro-2-nitropyridine 11 (300 mg, 1.36 mmol) in ethanol solvent (20 ml) heated and stirred at boiling under reflux for 1 hour. The reaction mixture is evaporated to dryness. The crude product was purified by silica chromatography (0-100% EtOAc/isohexane) to give the title compound 27 as a yellow gum (485 mg, 94%). R _t 1.17 min (method 2b) m/z 362/364 [M+H]+ (ER+), 1H NMR (DMSO-dQ: δ, ppm 8.24 (t, J=6.0 Hz, 1H), 8.05 (d, J=1.9 Hz, 1H), 7.87 (d, J=1.9 Hz, 1H), 5.19 (t, J=5.2 Hz, 1H), 4.08 (kvd, J=7.1 Hz, 2.2 Hz, 2H), 3.66-3.60 (m, 3H), 3.54 (dd, J=10.6 Hz, 5.0 Hz, 1H), 1.19-1.16 (m, 3H).

Step 4. 3-((5-Bromo-2-nitropyridin-3-yl)amino)-2-(hydroxymethyl)-2-methylpropanoic acid (compound 28).

A mixture of ethyl 3-((5-bromo-2-nitropyridin-3-yl)amino)-2-(hydroxymethyl)-2-methylpropanoate 27 (480 mg, 1.38 mmol) and lithium hydroxide monohydrate (49.5 mg, 2.07 mmol) in a solvent mixture of THF (20 ml) and water (5 ml) was stirred for 1 hour at RT. The mixture was acidified to pH ~3 by adding aqueous 1 M HCl and the mixture was evaporated to dryness to give the title compound 28 (501 mg, quant. yield), which was used in the next step without further purification. R _t 0.59 min (method 2b) 332/334 [MH] ^- (ER ^- ).

Step 5. 3-((2-Amino-5-bromopyridin-3-yl)amino)-2-(hydroxymethyl)-2-methylpropanoic acid (compound 29).

Mixture of 3-((5-bromo-2-nitropyridin-3-yl)amino)-2-(hydroxymethyl)-2-methylpropanoic acid 28 (460 mg, 1.38 mmol), iron powder (615 mg, 11.01 mmol) and ammonium chloride (147 mg, 2.75 mmol) in a solvent mixture of ethanol (30 ml) and water (8 ml) is heated and stirred at reflux for 1 hour. The reaction mixture is filtered through a pad of Celite®, and the filtrate is evaporated to dryness. The residue was triturated with ice water (10 mL), the solid was collected and dried in vacuo to give the title compound 29 as a gray solid (180 mg, 41% yield). R _t 0.71 min (method 1b) m/z 304/306 [M+H]+ (ER+); 302/304 [MH] ^- (ER ^- ), 1H NMR (400 MHz, DMSO-dQ: δ, ppm 7.32 (s, 1H), 7.17 (s, 2H), 6.79 (s , 1H), 5.78 (s, 2H), 4.79 (d, J=59.7 Hz, 2H), 3.54 (kv, J=10.6 Hz, 2H), 3.25-3 .12 (m, 2H), 1.13 (s, 3H).

Step 6. 8-Bromo-3-(hydroxymethyl)-3-methyl-2,3-dihydro-1H-pyrido[2,3-b][1,4]diazepin-4(5H)-one (compound 30) .

To a solution of 3-((2-amino-5-bromopyridin-3-yl)amino)-2-(hydroxymethyl)-2-methylpropanoic acid 29 (165 mg, 0.54 mmol) and DIPEA (284 μl, 1.63 mmol) in DMF (5 ml) add GATU (309 mg, 0.81 mmol) and the mixture is stirred for 1 hour. The reaction mixture is evaporated to dryness and the crude product is purified by silica chromatography (0-100% EtOAc/isohexane) to give the title compound 30 as a tan solid (31 mg, 20% yield). R _t 1.33 min (method 1b) m/z 286/288 [M+H]+ (ER+), ¹ H NMR (400 MHz, DMSO-0 ₆ ): δ, ppm 9.51 (s , 1H), 7.67 (d, J=2.1 Hz, 1H), 7.25 (d, J=2.1 Hz, 1H), 6.38 (t, J=4.5 Hz, 1H ), 4.79 (t, J=5.5 Hz, 1H), 3.60 (dd, J=10.5 Hz, 5.7 Hz, 1H), 3.44 (dd, J=10.6 Hz, 5.3 Hz, 1H), 3.20 (dd, J=13.8 Hz, 3.7 Hz, 1H), 3.07 (dd, J=13.8 Hz, 5.3 Hz, 1H ), 1.08 (s, 3H).

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Br

OH nbu ₄ ci, 1,4-dioxane, so °c

Step 7. (E)-3-(3-(Hydroxymethyl)-3-methyl-4-oxo-2,3,4,5-tetrahydro-III-pyrido[2,3-b][1,4]diazepine -8-yl)-N-methyl-N-((3-methylbenzofuran-2-yl)methyl)acrylamide (compound 31).

Na-methyl-N-((3-methylbenzofuran-2-yl)methyl)acrylamide 9 (61.7 mg, 0.27 mmol), 8-bromo-3-(hydroxymethyl)-3-methyl-2 is loaded into the reaction tube ,3-dihydro-1H-pyrido[2,3-b][1,4]diazepin4(5H)-one 30 (70 mg, 0.25 mmol), tetrabutylammonium chloride hydrate (7.24 mg, 0.02 mmol ), [P(tBu)3]Pd(crotyl)Cl (Pd-162) (Johnson Matthey) (9.77 mg, 0.02 mmol). The reaction tube was flushed with nitrogen for 5 min, 1,4-dioxane (10 mL) and N-cyclohexyl-N-methylcyclohexanamine (0.11 mL, 0.49 mmol) were added and the reaction mixture was purged with nitrogen for an additional 5 min. The mixture is heated to 80°C for 30 minutes. The reaction mixture was evaporated to dryness and the crude product was purified by chromatography on silica (0-10% MeOH/DCM) to give the title compound 31 as a yellow solid (71.2 mg, 66% yield). Rt 1.85 min (method 1b) m/z 435 [M+H] ⁺ (ER) ⁺ , 1H NMR (400 MHz, DMSOA 363 K): δ, ppm 8.88 (s, 1H), 7 .95 (d, J=2.0 Hz, 1H), 7.58-7.54 (m, 1H), 7.49-7.36 (m, 3H), 7.32-7.22 (m , 2H), 7.17-7.04 (m, 1H), 5.90 (s, 1H), 4.84 (s, 2H), 3.67 (d, J=10.5 Hz, 1H) , 3.51 (d, J=10.6 Hz, 1H), 3.27 (d, J=13.9 Hz, 1H), 3.15-3.07 (m, 4H), 2.28 ( s, 3H), 1.15 (s, 3H).

Step 8. Chiral resolution of compound 31.

Chiral separation method: apparatus: Isolera (Biotage). Chiralpak® IA (Daicel Ltd.) glass column (20 µm; 250 mmx25 mm), flow rate 40 ml min ^-1 , eluting with acetonitrile/ethanol (8/2 v/v). UV detection at 254 nm. Temperature: 30°C. Running time: 25 min. Samples are loaded after filtration through a 0.2 μm PTFE filter directly into the column. Injected quantity: 46.4 mg 31 in 10 ml eluent (final filtration through 0.2 µm PTFE filter).

(S,E)-3-(3-(Hydroxymethyl)-3-methyl-4-oxo-2,3,4,5-tetrahydro-1H-pyrido[2,3-b][1,4]diazepine- 8yl)-N-methyl-N-((3-methylbenzofuran-2-yl)methyl)acrylamide (compound 32). Pure fractions of the first enantiomer are collected and concentrated. The first enantiomer is obtained as a yellow solid (m=12.95 mg; chiral purity: 99.66%). R _t 1.53 min (method 1c) m/z 435.5 (M+H)+ (ER ⁺ ). The stereochemistry of the resulting enantiomer is determined arbitrarily.

(R,E-3-3-(3-(Hydroxymethyl)-3-methyl-4-oxo-2,3,4,5-tetrahydro-1H-pyrido[2,3-b][1,4]diazepine -8yl)-N-methyl-N-((3-methylbenzofuran-2-yl)methyl)acrylamide (compound 33) The pure fractions of the second enantiomer were collected and concentrated the first enantiomer was obtained as a yellow solid (m=8.97). mg; chiral purity: 98.20%). Rl 1.53 min (method 1c) m/z 435.5 [M+H] ⁺ (ER ⁺ ).

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Example 7. Synthesis of (E)-3-(3-(hydroxymethyl)-3-methyl-4-oxo-2,3,4,5-tetrahydro-1H-pyrido[2,3b][1,4]diazepine- 8-yl)-U-methyl-U-((2-methylbenzofuran-3-yl)methyl)acrylamide (compound 39) as well as two enantiomers N,E)-3-(3-(hydroxymethyl)-3-methyl- 4-oxo-2,3,4,5-tetrahydro-III-pyrido[2,3b][1,4]diazepin-8-yl)-U-methyl-U-((2-methylbenzofuran-3-yl) methyl)acrylamide (compound 40) and (R,E-3(3-(hydroxymethyl)-3-methyl-4-oxo-2,3,4,5-tetrahydro-1H-pyrido[2,3-b][ 1,4]diazepin-8-yl)-N-methyl-N((2-methylbenzofuran-3-yl)methyl)acrylamide (compound 41).

Reaction conditions: a) Cl ₂ CHOMe, TiCl ₄ , DCM; b) PhCHNHMe, Na(OAc) ₃ BH, EDC; c) Pd/C, H ₂ , MeOH, aq. HCl; d) Acryloyl chloride, TEA, THF; e) Pd-162, NCy ₂ Me, NBu ₄ Cl, 1,4-dioxane, 80°C; f) chiral separation.

CI ₂ CHOMe,

TiCI ₄ , dxm

Step 1. 2-Methylbenzofuran-3-carbaldehyde (compound 35).

To a solution of dichloro(methoxy)methane (5.1 ml, 56.7 mmol) and 2-methylbenzofuran 34 (5.0 g, 37.8 mmol) in DCM (100 ml) stirred at 0°C, add tin chloride dropwise (GU) (1 M in DCM) (60.5 mL, 60.5 mmol) for 30 min. After addition is complete, the mixture is warmed to RT for 30 min, then poured into ice-cold saturated sodium hydrogen carbonate solution (500 ml). The mixture is extracted into DCM (2x100 ml) and the organics are separated and dried. Filtration and evaporation gave the crude product, which was purified by silica chromatography (0-50% EtOAc/isohexane) to give the title compound 35 as a yellow solid (5.30 g, 86%), 1H NMR (400 MHz, CDCl ₃ ): δ, ppm 10.16 (s, 1H), 8.06-8.01 (m, 1H), 7.41-7.35 (m, 1H), 7.34-7, 23 (m, 2H), 2.70 (s, 3H).

_PhCH2NHMe

Na(OAc) ₃ BH, DCE /

Ph

Step 2. N-Benzyl-U-methyl-1-(2-methylbenzofuran-3-yl)methanamine (compound 36).

Sodium triacetoxyborohydride (1, 99 g, 9.37 mmol) and the mixture was stirred for 72 hours. The reaction mixture was washed with saturated sodium bicarbonate solution (20 ml) and dried over sodium sulfate. Filtration and evaporation gave the title compound 36 as a pale yellow oil (1.60 g, 94% yield), which was used without further purification. 1H NMR (400 MHz, DMSO^): δ, ppm 7.63-7.58 (m, 1H), 7.48-7.43 (m, 1H), 7.33 (d, J=4 .8 Hz, 4H), 7.28-7.20 (m, 3H), 3.56 (s, 2H), 3.52 (s, 2H), 2.42 (s, 3H), 2.08 (c, 3H).

Step 3. Namethyl-1-(2-methylbenzofuran-3-yl)methanamine hydrochloride (compound 37).

A mixture of N-benzyl-U-methyl-1-(2-methylbenzofuran-3-yl)methanamine 36 (1.60 g, 6.03 mmol) and Pd-C 87 l 5% on carbon (0.64 g, 6. 03 mmol) in methanol (20 ml) is acidified to pH 1 with 1 M hydrochloric acid, hydrogenated at 5 bar and stirred at RT for 18 hours. The catalyst is removed by filtration, and

- 55 046887 the filtrate is evaporated to dryness to obtain the title compound 37 as a white solid (737 mg, 56%), ¹ H NMR (400 MHz, DMSO-06): δ, ppm 9.24 (s, 2H ), 7.89-7.80 (m, 1H), 7.59-7.49 (m, 1H), 7.34-7.23 (m, 2H), 4.24 (s, 2H), 2.57 (s, 3H), 2.55 (s, 3H).

Step 4. ^Methyl^-((2-methylbenzofuran-3-yl)methyl)acrylamide (compound 38).

To a suspension of Namethyl-1-(2-methylbenzofuran-3-yl)methanamine hydrochloride 37 (300 mg, 1.42 mmol) and triethylamine (600 μl, 4.25 mmol) in dry THF (10 ml) add acryloyl chloride (154 mg, 1.70 mmol) dropwise at RT for 15 minutes. The mixture is stirred for 1 hour and then poured into water (30 ml). The organic solvent is removed by rotary evaporation to obtain a solid. The solid was collected by filtration, washed with water (10 mL) and dried to give the title compound 38 (316 mg, 95%) as a colorless solid. Rt 1.94 min (method 1b) m/z 230 [M+H]+ (ER ⁺ ).

(E)-3 -(3 -(G hydroxymethyl)-3 -methyl-4-oxo-2,3,4,5-tetrahydro-1 H-pyrido[2,3 Stage

5.

b][1,4]diazepin-8-yl)-Nmethyl-N((2-methylbenzofuran-3-yl)megyl)acrylamide (compound 39).

Namethyl-N-((2-methylbenzofuran-3-yl)methyl)acrylamide 38 (72.0 mg, 0.32 mmol), 8-bromo-3-(hydroxymethyl)-3-methyl-2 is loaded into the reaction tube ,3-dihydro-1H-pyrido[2,3-b][1,4]diazepin4(5H)-one 30 (90.0 mg, 0.32 mmol), tetrabutylammonium chloride hydrate (9.3 mg, 0.031 mmol ), [P(tBu) ₃ ]Pd(crotyl)Cl (Pd-162) (12.6 mg, 0.03 mmol). The reaction tube is flushed with nitrogen for 5 minutes. 1,4-Dioxane (10 ml) and N-cyclohexyl-N-methylcyclohexanamine (0.14 ml, 0.63 mmol) were added and the reaction mixture was purged with nitrogen for a further 5 minutes. The mixture is heated to 80°C for 30 minutes. The reaction mixture was evaporated to dryness and the crude product was purified by chromatography on silica (0-10% MeOH/DCM) to give the title compound 39 as a yellow solid (94.3 mg, 68% yield). R _t 1.21 min (method 2a) m/z 435 [M+H] ⁺ (ER ⁺ ), ¹ H NMR (400 MHz, DMSO-dg, 363 K): δ, ppm 8.88 (s , 1H), 7.95 (d, J=2.0 Hz, 1H), 7.58-7.36 (m, 4H), 7.28-7.04 (m, 3H), 5.88 ( t, J=4.7 Hz, 1H), 4.77 (s, 2H), 4.45 (t, J=5.5 Hz, 1H), 3.67 (dd, J=10.6 Hz, 5.7 Hz, 1H), 3.54-3.48 (m, 1H), 3.26 (dd, J=13.9 Hz, 3.9 Hz, 1H), 3.11 (dd, J= 13.9 Hz, 5.3 Hz, 1H), 3.01 (shs, 3H), 1.14 (s, 3H). Note: The methyl group on benzofuran is below the solvent peak.

Step 6. Chiral resolution of compound 39.

Chiral separation method: apparatus: Isolera (Biotage). Chiralpak® IA (Daicel Ltd.) glass column (20 µm; 250 mmx25 mm), flow rate 40 ml min ^-1 , eluting with acetonitrile/ethanol (8/2 v/v). UV determination at 254 nm. Temperature: 30°C. Running time: 25 min. Samples are loaded after filtration through a 0.2 μm PTFE filter directly into the column. Injected quantity: 56.1 mg 39 in 10 ml eluent (final filtration through 0.2 µm PTFE filter).

- 56 046887 (S,E)-3 -(3 -(G hydroxymethyl)-3 -methyl-4-oxo-2,3,4,5-tetrahydro-1H-pyrido[2,3-b] [1, 4]diazepin-8yl)-N-methyl-N-((2-methylbenzofuran-3-yl)methyl)acrylamide (compound 40).

Pure fractions of the first enantiomer are collected and concentrated. The first enantiomer is obtained as a yellow solid (m=21.11 mg; chiral purity: 98.82%). R _t 1.50 min (standard) m/z 435.5 [M+H]+ (ER+). The stereochemistry of the resulting enantiomer is determined arbitrarily.

(R,E-3 -(3 -(G hydroxymethyl)-3 -methyl-4-oxo-2,3,4,5-tetrahydro-1H-pyrido[2,3 -b][1,4]diazepine- 8-yl)Ν-μοτηλ-Ν-((2-methylbenzofuran-3-yl)methyl)acrylamide (compound 41).

Pure fractions of the second enantiomer are collected and concentrated. The first enantiomer is obtained as a yellow solid (m=18.22 mg; chiral purity: 99.49%). R _t 1.50 min (standard) m/z 435.5 [M+H]+ (ER+). The stereochemistry of the resulting enantiomer is determined arbitrarily.

Example 8. Synthesis of (NE-3-(3-Hydroxy-3-methyl-4-oxo-2,3,4,5-tetrahydro-1H-pyrido[2,30][1,4]diazepine-8- yl)-N-methyl-N-((3-methylbenzofuran-2-yl)methyl)acrylamide (compound 49) and (S,E)3-(3-hydroxy-3-methyl-4-oxo-2,3 ,4,5-tetrahydro-1H-pyrido[2,3^][1,4]diazepin-8-yl)-N-methyl-N-((3methylbenzofuran-2-yl)methyl)acrylamide (compound 50).

General synthesis scheme.

Reaction conditions: a) 5-bromo-3-fluoro-2-nitropyridine 11, K2CO3, reflux; b) Fe, AcOH, 90°C; c) LiOH, THF:H ₂ O, RT; d) GATU, Huenig base, DMF, CT; e) Chiralpack IA, EUN/heptane/SHC1 ₃ ; f) Pd-162, NBu ₄ Cl, Cy ₂ NMe, 80°C.

Step 1. Methyl 3-((5-bromo-2-nitropyridin-3-yl)amino)-2-hydroxy-2-methylpropanoate (compound 43).

To a stirred solution of 5-bromo-3-fluoro-2-nitropyridine 11 (0.33 g, 1.47 mmol) in THF (5 ml) add methyl 3-amino-2-hydroxy-2-methylpropanoate hydrochloride 42 (enamine) (0.25 g, 1.47 mmol), then potassium carbonate (0.41 g, 2.95 mmol). The reaction mixture is stirred at reflux for 2 hours. The reaction mixture is cooled to room temperature. The solvent was removed in vacuo to give the title compound 43 as a yellow oil (0.48 g, 94% yield). R _t 1.03 min (method 2a) m/z 334/336 [M+H]+(ER+).

44

Step 2. 3-((2-Amino-5-bromopyridin-3-yl)amino)-2-hydroxy-2-methylpropanoate (compound 44).

To a stirred solution of methyl 3-((5-bromo-2-nitropyridin-3-yl)amino)-2-hydroxy-2methylpropanoate 43 (0.48 g, 1.44 mmol) in EtOH (6 ml) add acetic acid ( 1.64 ml, 28.7 mmol), then iron powder (0.80 g, 14.4 mmol). The reaction mixture is stirred at 90°C for 30 minutes. The reaction mixture is cooled to room temperature and neutralized to pH 8 with solid NaHCO ₃ . The resulting reaction mixture was diluted with H2O (10 ml) and EtOAc (10 ml). The aqueous phase is separated and extracted with EtOAc (2x10 ml). The combined organic phases are washed with H2O (10 ml), passed through a hydrophobic frit, and concentrated in vacuo to give the title compound 44 (0.43 g, 50%). R _t 0.58 min (method 2a) m/z 304/306 [M+H]+ (ER+).

- 57 046887

Step 3. 3-((2-Amino-5-bromopyridin-3-yl)amino)-2-hydroxy-2-methylpropanoic acid (compound 45).

To a stirred solution of methyl 3-((2-amino-5-bromopyridin-3-yl)amino)-2-hydroxy-2methylpropanoate 44 (0.35 g, 1.15 mmol) in THF (1.5 ml) add a solution LiOH (0.14 g, 5.8 mmol) in H2O (1.5 ml). The reaction mixture is stirred at room temperature overnight. The solvent was evaporated to dryness to obtain the title compound 45 as a colorless solid (320 mg, 34% yield), which was used in the next step without further purification. R _t 0.73 min (method 2a) m/z 290/292 [M+H]+ (ER ⁺ ).

Step 4. 8-Bromo-3-hydroxy-3-methyl-1,2,3,5-tetrahydro-4H-pyrido[2,3-b][1,4]diazepin-4-one (compound 46).

DIPEA (0 .58 ml, 3.3 mmol), then GATU (0.63 g, 1.65 mmol). The reaction mixture was stirred at room temperature for 1 hour. Water (4 ml) was added. The aqueous phase is separated and extracted with EtOAc (2x5 ml). The combined organic phases are washed with H ₂ O (3x5 ml), passed through a hydrophobic frit, and concentrated in vacuo to give the title compound 46 as a colorless solid (41 mg, 14% yield). R _t 0.83 min (method 2a) m/z 272/274 [M+H] ⁺ (ER ⁺ ), ¹ H NMR (400 MHz, DMSOC): δ, ppm 9.92 (s, 1H) , 7.65 (d, J=2.1 Hz, 1H), 7.24 (d, J=2.1 Hz, 1H), 6.50 (t, J=4.6 Hz, 1H), 5 .34 (s, 1H), 3.283.05 (m, 2H), 1.22 (s, 3H). Step 5. Chiral resolution of racemate 46.

Enantiomers are separated by chiral prep. HPLC using method 3 a. Chirality is determined arbitrarily.

(R)-8-Bromo-3-hydroxy-3-methyl-1,2,3,5-tetrahydro-4H-pyrido[2,3-b][1,4]diazepin-4-one (compound 47) . First isomer eluted: R _t 6.4 min (method 4a); R _t 0.83 min (method 2a) m/z 272/274 [M+H] ⁺ (er ⁺ ).

(S)-8-Bromo-3-hydroxy-3-methyl-1,2,3,5-tetrahydro-4H-pyrido[2,3-b][1,4]diazepin-4-one (compound 48) . Second isomer eluted: R _t 11.7 min (method 4a); R _t 0.83 min (method 2a) m/z 272/274 [M+H] ⁺ (er ⁺ ).

DU/NUI, Ί,4-dioxane 47 ⁴ 49

- 58 046887

Stage 6a. ^,E-3-(3-Hydroxy-3-methyl-4-oxo-2,3,4,5-tetrahydro-III-pyrido[2,3-b][1,4]diazepin-8yl)-U -methyl-U-((3-methylbenzofuran-2-yl)methyl)acrylamide (compound 49).

Namethyl-U-((3-methylbenzofuran-2-yl)methyl)acrylamide 9 (22 mg, 0.1 mmol), (R)-8-bromo-3-hydroxy-3-methyl-2 is loaded into the reaction tube ,3-dihydro-1H-pyrido[2,3-b][1,4]diazepin-4(5H)-one 47 (27 mg, 0.1 mmol), tetrabutylammonium chloride hydrate (3 mg, 0.01 mmol ), [P(tBu) ₃ ]Pd(crotyl)Cl (Pd162) (4 mg, 0.01 mmol). The tube is then flushed with nitrogen for 5 minutes. 1,4-Dioxane (2 ml) and Mcyclohexyl-Mmethylcyclohexanamine (42 µl, 0.20 mmol) were added and the reaction mixture was purged with nitrogen for a further 5 min. The mixture is heated to 80°C for 3 hours and cooled to room temperature. The solvent is evaporated to dryness. The crude product was purified by silica chromatography (0-10% MeOH/DCM) to give the title compound 49 as a yellow solid (20 mg, 48%). R _t 19 min (method 4a). R _t 1.23 min (method 2a) m/z 421 [M+H]+ (ER ⁺ ), ¹ H NMR (400 MHz, DMSO^): δ, ppm 9.37 (s, 1H), 7.95 (d, J=1.9 Hz, 1H), 7.60-7.53 (m, 1H), 7.49-7.45 (m, 1H), 7.42 (d, J= 15.4 Hz, 1H), 7.36 (d, J=2.0 Hz, 1H), 7.30-7.24 (m, 2H), 7.18-7.07 (m, 1H), 6.06 (s, 1H), 4.92 (s, 1H), 4.84 (s, 2H), 3.20 (dd, J=6.7 Hz, 4.7 Hz, 2H), 3, 10 (s, 3H), 2.28 (s, 3H), 1.29 (s, 3H).

50

Stage 6b. (S,E)-3-(3-Hydroxy-3-methyl-4-oxo-2,3,4,5-tetrahydro-1H-pyrido[2,3-b][1,4]diazepin-8yl) -U-methyl-U-((3-methylbenzofuran-2-yl)methyl)acrylamide (compound 50).

Namethyl-U-((3-methylbenzofuran-2-yl)methyl)acrylamide 9 (23 mg, 0.1 mmol), (S)-8-bromo-3-hydroxy-3-methyl-2 is loaded into the reaction tube ,3-dihydro-1H-pyrido[2,3-b][1,4]diazepin-4(5H)-one 48 (27 mg, 0.1 mmol), tetrabutylammonium chloride hydrate (3 mg, 0.01 mmol ), [P(tBu) ₃ ]Pd(crotyl)Cl (Pd162) (4 mg, 0.01 mmol). The tube is then flushed with nitrogen for 5 minutes. 1,4-Dioxane (2 mL) and N-cyclohexyl-N-methylcyclohexanamine (43 μL, 0.20 mmol) were added and the reaction mixture was purged with nitrogen for a further 5 min. The mixture is heated to 80°C for 3 hours and cooled to room temperature. The solvent is evaporated to dryness. The crude product was purified by silica chromatography (0-10% MeOH/DCM) to give the title compound 50 as a yellow solid (34 mg, 80% yield). R _t 33 min (method 4a). R _t 1.23 min (method 2a) m/z 421 [M+H]+ (ER+), 1H NMR (400 MHz, DMSO-dQ: δ, ppm 9.37 (s, 1H), 7 .95 (d, J=1.9 Hz, 1H), 7.60-7.53 (m, 1H), 7.49-7.45 (m, 1H), 7.42 (d, J=15 .4 Hz, 1H), 7.36 (d, J=2.0 Hz, 1H), 7.30-7.24 (m, 2H), 7.18-7.07 (m, 1H), 6 .06 (s, 1H), 4.92 (s, 1H), 4.84 (s, 2H), 3.20 (dd, J=6.7, 4.7 Hz, 2H), 3.10 ( s, 3H), 2.28 (s, 3H), 1.29 (s, 3H).

Example 9. Synthesis of 8-((E)-3-oxo-3-((3aS,6aR)-5-(4-(pyrimidin-2-yloxy)phenyl)-3,3a,4,6atetrahydrocyclopentyl[c]pyrrole -2(1H)-yl)prop-1-en-1-yl)-1,2,3,5-tetrahydro-4H-pyrido[2,3-b][1,4]diazepin-4-one ( connection 60).

General synthesis scheme.

Reaction conditions: a) Cs ₂ CO ₃ , NMP, 80°C, 2 h; b) Bis(pinacolato)diborone 54, Pd(dppf)Cl ₂ , AcOK, 1,4-dioxane; c) LHMDS, Comins reagent, THF, -78°C to RT; d) Na ₂ CO ₃ , Pd(PPh ₃ ) ₄ , H ₂ O, 1,4-dioxane, 90°C; e) TPA, DXM; f) DIPEA, GATU, DMF.

- 59 046887

CI but

Br Cs ₂ CO ₃ ,NMP, °C, 2 h

Br

Step 1. 2-(4-Bromophenoxy)pyrimidine (compound 53).

A mixture of 2-chloropyrimidine 51 (Alfa Aesar) (2 g, 17 mmol), 4-bromophenol 52 (Sigma-Aldrich) (3.6 g, 21 mmol) and Cs ₂ CO ₃ (17 g, 52 mmol) in U- methyl 2-pyrrolidone (50 ml) is heated at 80°C for 2 hours. The reaction mixture is cooled to room temperature and poured into water (200 ml). The mixture is extracted with EtOAc (2x100 ml). The organics are combined and washed with brine (2 x 100 ml), passed through a hydrophobic frit and concentrated in vacuo. The crude product was purified by chromatography (0-50% EtOAc/isohexane) to give the title compound 53 as a colorless crystalline solid (3.95 g, 90% yield). R _t 1.86 min (method 1a) m/z 251/253 [M+H]+ (ER ⁺ ), ¹ H NMR (400 MHz, CDCl ₃ ): δ, ppm 8.59 (d, J =4.7 Hz, 2H), 7.59-7.53 (m, 2H), 7.157.10 (m, 2H), 7.08 (t, J=4.8 Hz, 1H).

Pd(dppf)CI2j AcOK. 1,4-dioxane

Step 2. 2-(4-(4,4,5,5-Tetramethyl-1,3,2-dioxaborolan-2-yl)phenoxy)pyrimidine. Stage 2 (compound 55).

2-(4-bromophenoxy)pyrimidine mixture 53 (0.5 g, 2.0 mmol), 4,4,4',4',5,5,5',5'-octamethyl-2,2'-bi (1,3,2dioxaborolane) 54 (Fluorochem) (0.7 g, 2.8 mmol), potassium acetate (0.6 g, 6.0 mmol) and PdCl ₂ (dppf) (0.07 g, 0. 1 mmol) in 1,4-dioxane (20 ml) is stirred at 85°C for 16 hours. The reaction mixture is cooled to room temperature, filtered through Celite®, and concentrated in vacuo. The crude residue is partitioned between EtOAc (50 ml) and water (50 ml). The aqueous phase is extracted with EtOAc (2x50 ml). The combined organic layers were passed through a phase separator and evaporated to dryness to give crude product 55 as a dark brown solid (0.6 g, 2.0 mmol, qty). R _t 2.23 min (method 1a) m/z 299 [M+H] ⁺ (ER ⁺ ).

THF, -78 °C to rt

LHMDS, Comins reagent

Vos

Step 3. Preparation of tert-butyl (3aS,6aS)-5-(((trifluoromethyl)sulfonyl)oxy)-3,3a,4,6atetrahydrocyclopenta[c]pyrrole-2(1H)-carboxylate (compound 57).

To a stirred solution of tert-butyl 5-oxohexahydrocyclopenta[c]pyrrole-2(1H)carboxylate 56 (Ark Pharm) (3 g, 13.3 mmol) in THF (60 mL), slowly add a solution of LHMDS (1M in THF) (17 ml, 17.3 mmol) at -78°C. The reaction mixture was stirred at -78°C for 30 min, then a solution of Na(5-chloropyridin-2-yl)-1,1,1-trifluoro-Na((trifluoromethyl)sulfonyl)methanesulfonamide (7.3 g, 18.6 mmol) in THF (20 ml). The mixture is stirred for an additional 1 hour at -78°C, warmed to room temperature and stirred for 2 hours. The reaction mixture is concentrated in vacuo. The crude residue was purified by chromatography (0-50% EtOAc/isohexane) to give the title compound 57 as a pale yellow oil (4.4 g, 91% yield). Rt 1.29 min (method 1a) m/z 302 [M+H-tBu] ⁺ (ER ⁺ ). 1H NMR (400 MHz, CDCl ₃ ): δ, ppm 5.58 (d, J=1.9 Hz, 1H), 3.72 (dd, J=11.4 Hz, 8.5 Hz, 1H ), 3.53 (dd, J=11.8 Hz, 8.3 Hz, 1H), 3.40 (ddt, J=8.6 Hz, 6.0 Hz, 2.7 Hz, 2H), 3 .16 (dd, J=11.4, 6.5 Hz, 1H), 3.04-2.80 (m, 2H), 2.51-2.33 (m, 1H), 1.46 (s , 9H).

, Vos

,Vos

I Pd(PPh ₃ ) ₄ ,

MezSOz, H2O, 1,4-dioxane, 90'

Step 4. tert-Butyl (3aS,6aR)-5-(4-(pyrimidin-2-yloxy)phenyl)-3,3a,4,6a-tetrahydrocyclopenta[c]pyrrole-2(1H)-carboxylate (compound 58 ).

To a stirred suspension of (3aS,6aS)-tert-butyl 5-(((trifluoromethyl)sulfonyl)oxy)-3,3a,6,6atetrahydrocyclopenta^pyrrole^SDcarboxylate 57 (0.28 g, 0.77 mmol), 2- (4-(4,4,5,5-tetramethyl1,3,2-dioxaborolan-2-yl)phenoxy)pyrimidine 55 (0.30 g, 1.00 mmol), Pd(Ph ₃ P) ₄ (0, 09 g, 0.08 mmol) in 1,4-dioxane (20 ml) add a solution of Na ₂ CO ₃ (0.5 g, 5.0 mmol) in H2O (2.5 ml). The reaction mixture is stirred at 85°C for 16 hours. The reaction mixture is cooled to room temperature,

- 60 046887 filtered through Celite® and concentrated in vacuo. The crude residue is diluted with water (50 ml), extracted with EtOAc (2x50 ml). The combined organic layers are passed through a phase separator and evaporated to dryness. The crude product was purified by chromatography (0-100% EtOAc/isohexane) to give the title compound 58 as a clear, colorless oil (0.18 g, 60% yield). R _t 2.44 min (method 1a) m/z 324 [M+H-tBu]+ (ER ⁺ ). ¹ H NMR (400 MHz, CDCl ₃ ): δ, ppm, 8.59 (d, J=4.8 Hz, 2H), 7.50 (d, J=8.7 Hz, 2H), 7 ,21-7.15 (m, 2H), 7.06 (t, J=4.8 Hz, 1H), 6.03 (s, 1H), 3.72 (dd, J=10.7 Hz, 8.4 Hz, 1H), 3.61-3.47 (m, 3H), 3.14-2.93 (m, 3H), 2.62 (d, J=15.5 Hz, 1H), 1.47 (s, 9H).

Step 5. (3aS,6aR)-5-(4-(Pyrimidin-2-yloxy)phenyl)-1,2,3,3a,4,6a-hexahydrocyclopenta[c]pyrrole (compound 59).

To a stirred solution of (3aR,6aS)-tert-butyl 5-(4-(pyrimidin-2-yloxy)phenyl)-3,3a,6,6atetrahydrocyclopenta^pyrrole-3-carboxylate 58 (0.18 g, 0. 47 mmol) in DCM (1 ml) add TFA (1 ml). The reaction mixture is stirred at room temperature for 2 hours. The solvent is removed in vacuo. The resulting oil was taken up in MeOH (5 mL) and applied to an SCC column. The column was washed with methanol (5 mL) and the product was eluted with 10% methanolic ammonia (5 mL) to give the title compound 59 as a colorless crystalline solid (0.09 g, 69%). Rt 0.92 min (method 1a) m/z 280 [M+H]+ (ER ⁺ ).

6. 8-((E)-3-Oxo-3-((3aS,6aR)-5-(4-(pyrimidin-2-yloxy)phenyl)-3,3a,4,6a-tetrahydrocycloPenta step[c] pyrrol-2(1H)-yl)prop-1-en-1-yl)-1,2,3,5-tetrahydro-4H-pyrido[2,3-b][1,4]diazepin-4-one (compound 60).

To a stirred solution of (3aS,6aR)-5-(4-(pyrimidin-2-yloxy)phenyl)-1,2,3,3a,4,6ahexahydrocyclopenta-pyrrole 59 (0.09 g, 0.31 mmol) in DMF (2 ml) add (E)-3(4-oxo-2,3,4,5-tetrahydro-1H-pyrido[2,3-b][1,4]diazepin-8-yl)acrylic acid trifluoroacetate 6 (79 mg, 0.34 mmol) and Na-ethyl-N-isopropylpropan-2-amine (0.27 ml, 1.54 mmol). The reaction mixture was stirred at RT for 5 min and GATU (0.14 g, 0.37 mmol) was added. The reaction mixture was stirred at room temperature for 2 hours. H2O (1 ml) was added and the reaction mixture was sonicated. The solid is filtered and washed with H2O (2x2 ml). The filtrate is discarded. The crude solid was purified by chromatography (0-10% MeOH/DCM) to give the title compound 60 as a yellow solid (0.13 g, 79% yield). Rt 1.65 min (method 1a) m/z 495 [M+H]+ (ER+), ¹ H NMR (400 MHz, DMSO-dg): δ, ppm 9.74 (d, J=7 ,3 Hz, 1H), 8.64 (dd, J=4.8 Hz, 1.5 Hz, 2H), 7.98 (dd, J=9.3 Hz, 1.9 Hz, 1H), 7 .58-7.51 (m, 2H), 7.38-7.30 (m, 2H), 7.27 (td, J=4.8 Hz, 0.8 Hz, 1H), 7.21- 7.14 (m, 2H), 6.87 (dd, J=15.5 Hz, 9.4 Hz, 1H), 6.22-6.15 (m, 1H), 6.03 (d, J =14.4 Hz, 1H), 4.06-3.77 (m, 2H), 3.72-3.50 (m, 2H), 3.45-3.39 (m, 2H), 3, 24-2.84 (m, 3H), 2.69-2.55 (m, 3H).

Example 10. Preparation of (E)-N-methyl-N-((2-methylbenzofuran-3-yl)methyl)-3-(3'-oxo-2,3,3',4',5,6hexahydro-1Ή -spiro[pyran-4,2'-pyrido[2,3-b]pyrazin]-7'-yl)acrylamide (compound 64).

General synthesis scheme.

- 61 046887

Reaction conditions: a) K2CO3, THF; b) NH4C1, Fe, EtOH/water; c) Pd(OAc)2, tri-o-tolylphosphane, DIPEA in DMF/ACN.

Step 1. 4-((5-Bromo-2-nitropyridin-3-yl)amino)tetrahydro-2H-pyran-4-carboxylic acid (compound 62).

5-Bromo-3-fluoro-2-nitropyridine 11 (800 mg, 3.62 mmol, 1.0 eq) and 4-aminotetrahydro-2H-pyran-4carboxylic acid 61 (Combi-Blocks) (735 mg, 5, 06 mmol, 1.4 eq.) dissolved in THF (10 ml), then add K2CO3 (1 g). The resulting reaction mixture is heated to 90°C overnight in a sealed tube. UPLC analysis showed almost complete conversion of IM with the formation of the expected product (R _t = 1.11 min; ER = 348.1). The solvent is removed in vacuo and the residue is dissolved in water/EtOAc. The aqueous phase is acidified with aq. 1 M HCl to pH 1 and extracted twice with EOAc. The combined organic layers are washed with brine, dried over MgSO ₄ and concentrated to dryness. The resulting yellow solid is diluted with EtOAc. The resulting precipitate was filtered off to give product 62 as a yellow solid with 99% purity (535 mg, 42% yield). The filtrate was concentrated to dryness to give the less pure product 62 as a brown solid with a purity of 93% (550 mg, 43.8% yield). Both batches are used in the next step without purification. R _t 1.09 min (method 1c) m/z 346.1/348.1 [M+H]+ (ER ⁺ ) (isotope effect).

63

Step 2. 7'-Bromo-1',2,3,4',5,6-hexahydro-3Ή-spiro[pyran-4,2'-pyrido[2,3-b]pyrazin]-3'-one (compound 63).

To a stirred solution of 4-((5-bromo-2-nitropyridin-3-yl)amino)tetrahydro-2H-pyran-4carboxylic acid 62 (535 mg, 1.5 mmol, 1.0 eq.), NH ₄ Cl ( 165 mg, 3.1 mmol, 2.0 eq) and iron (170 mg, 3.1 mmol, 2.0 eq) suspended in EtOH/Water (1:1 v/v, 10 ml) . The resulting suspension is heated to 80°C for 1.5 hours. UPLC analysis showed the formation of a product (Rt = 1.09 min; ER ⁺ = 300.1) with complete conversion of the starting material. The reaction mixture is diluted with EtOAc (and a small amount of MeOH), filtered through a 0.22 µM filter and washed with acetonitrile. The solution is concentrated to dryness and diluted with AcOEt and washed with water. The organic phase is dried (MgSO ₄ ), filtered and concentrated to dryness. The residue is purified on silica gel using a SNAP Ultra 25 g cartridge and DCM/MeOH 99:1 to 95:5 v/v. as an eluent. Fractions containing the product are combined to give 7'-bromo-1',2,3,4',5,6-hexahydro-3Ή-spiro[pyran-4,2'-pyrido[2,3-b]pyrazine] -3'-one 63 as a gray-green powder with a purity of 99% (220 mg, 48% yield). UPLC-MS m/z: 300.1 [M+H] ⁺ (ER ⁺ ). ¹ H NMR (400 MHz, DMSO-ph): δ, ppm 10.94 (s, 1H), 7.69 (d, J=2.1 Hz, 1H), 7.33 (d, J= 2.0 Hz, 1H), 6.77 (s, 1H), 3.75-3.70 (m, 4H), 2.00-1.90 (m, 2H), 1.46 (d, J =13.6 Hz, 2H).

Step 3. (E)-N-Methyl-N-((2-methylbenzofuran-3-yl)methyl)-3-(3'-oxo-2,3,3',4',5,6-hexahydro- 1Ήspiro[pyran-4,2'-pyrido[2,3-b]pyrazin]-7'-yl)acrylamide (compound 64).

7'-bromo-1',2,3,4',5,6-hexahydro-3Ή-spiro[pyran-4,2'pyrido[2,3-b]pyrazine]-3' is loaded into an oven-dried test tube -one 63 (50 mg, 0.16 mmol), add anhydrous DMF (0.9 ml) and acetonitrile (0.1 ml). The reaction mixture is degassed with nitrogen for 5 minutes. \-Methyl-\-((2methylbenzofuran-3-yl)methyl)acrylamide 38 (58 mg, 0.25 mmol) and DIPEA (109 µl) were then added and the mixture was degassed a second time for 10 min. Finally, Pd(OAc) ₂ (0.4 mg) and tri-o-tolylphosphane (1.0 mg) are added in one portion and nitrogen is bubbled through the mixture for 1 min, and the solution is heated at 100°C overnight . The reaction mixture is cooled to RT and filtered over a Celite® pad. The cake is washed with MeOH/AcOEt and the filtrate is concentrated under high vacuum to give a crude brown solid (250 mg), which is purified by Isolera (DCM 100% to DCM/AcOEt 0/100%). All fractions containing product were evaporated to obtain the title molecule 64 as a white solid with 97% purity (46 mg, 0.10 mmol, 64% yield). R _t 1.55 min (method 1c). UPLC-MS

- 62 046887 m/z: 447.1 [M+H]+ (ER ⁺ ). 1H NMR (400 MHz, DMSO-06): δ, ppm 10.97 (s, 1H), 8.00 (s, 1H), 7.96 (s, 1H), 7.57 (d, J =7.3 Hz, 1H), 7.52 (d, J=15.4 Hz, 1H), 7.49-7.44 (m, 2H), 7.27-7.15 (m, 2H) , 6.99 (d, J=15.3 Hz, 1H), 6.63-6.56 (m, 1H), 4.89 (s, 2H), 4.75 (s, 2H), 3, 79-3.66 (m, 4H), 3.05 (s, 2H), 2.83 (s, 1H), 1.99-1.89 (m, 2H), 1.46 (d, J= 14.3 Hz, 2H), 1.24 (s, 3H) (rotamers).

Example 11. Synthesis of (E)-N-((7-Amino-2-methylbenzofuran-3-yl)methyl)-N-methyl-3-(4-oxo-2,3,4,5tetrahydro-1H-pyrido [2,3-b][1,4]diazepin-8-yl)acrylamide (compound 75).

General synthesis scheme.

Reaction conditions: a) Ethyl 2-bromopropanoate 66, K ₂ CO ₃ , MeCN, reflux, then aq. NaOH, THF, reflux; b) NaOAc, Ac ₂ O, reflux; c) dichloro(methoxy)methane, SnCl ₄ , 1M in DCM, DCM, 0°C to RT; d) 1-(4-methoxyphenyl)-Nmethylmethanamine 70, Na(OAc) ₃ BH, DCE; e) Pd ₂ (dba) ₃ , Xantphos, benzophenonymine 72, Cs ₂ CO ₃ , PhCh ₃ , 100°C; f) 1-chloroethyl chloroformate, DCM, 0°C - rt; g) GATU, DIPEA, DMF, kg.

Step 1. 2-(2-Bromo-6-formylphenoxy)propanoic acid (compound 67).

K2CO3 (13.8 g, 99 mmol) was added in one portion to a stirred solution of 3-bromo-2hydroxybenzaldehyde 65 (10 g, 49.7 mmol) and ethyl 2-bromopropanoate 66 (5.9 mL, 45.2 mmol) in MeCN (80 ml), and the reaction mixture is heated to reflux for 3 hours. The reaction mixture is cooled to rt, filtered to remove K2CO3 and then concentrated in vacuo. The resulting residue was dissolved in THF (50 ml) and NaOH solution (2M in H2O, 57 ml, 113 mmol) was added. The mixture is heated to reflux for 2 hours, then cooled to RT and concentrated in vacuo. The remaining aqueous product is acidified to pH 1 by adding concentrated HCl dropwise and the product precipitates. The product was collected by filtration and dried by azeotropic distillation with MeCN (2x50 ml) to give the desired product 67 as an orange oil which crystallized on standing to give a whitish solid (13.5 g, 99% yield). R _t 1.19 min (method 2a) m/z 273/275 [M+H] ⁺ (ER ⁺ ), ¹ H NMR (400 MHz, DMSO-06): δ, ppm 13.23 (s, 1H), 10.41 (d, J=0.8 Hz, 1H), 7.97 (dd, J=7.9, 1.7 Hz, 1H), 7.74 (dd, J=7.7 Hz, 1.7 Hz, 1H), 7.25 (td, J=7.8 Hz, 0.9 Hz, 1H), 4.93 (kv, J=6.8 Hz, 1H), 1.60 (d, J=6.8 Hz, 3H).

- 63 046887

Ac ₂ O, NaOAc bp. arr. cold;

68

Step 2. 7-Bromo-2-methylbenzofuran (compound 68).

A mixture of 2-(2-bromo-6-formylphenoxy)propanoic acid 67 (13 g, 48 mmol) and sodium acetate (39 g, 48 mmol) in acetic anhydride (70 ml) was heated to reflux for 2 hours. The mixture is cooled to RT, then poured into ice water (800 ml). The mixture was then extracted with DCM (3x300 ml) and the combined organic layers were washed with NaOH (2M aq, 2x200 ml), then brine (200 ml). The organic layer is dried by passing through a phase separator, then concentrated in vacuo. The crude product was purified by column chromatography (5-10% EtOAc/isohexane) to give the desired product 68 as a colorless oil (6.72 g, 66% yield). R _t 1.67 min (method 2a) m/z not observed. ^1H NMR (400 MHz, DMSO^ ₆ ): δ, ppm 7.53 (dd, J=7.7 Hz, 1.0 Hz, 1H), 7.43 (dd, J=7.8 Hz , 1.0 Hz, 1H), 7.13 (t, J=7.8 Hz, 1H), 6.71 (kv, J=1.1 Hz, 1H), 2.48 (d, J=1 ,1 Hz, 3H).

Step 3. 7-Bromo-2-methylbenzofuran-3-carbaldehyde (compound 69).

Tin chloride (TC) (38 mL, 38 mmol, 1 M in DCM) is added dropwise over ~30 min to a stirred solution of dichloro(methoxy)methane (3.2 mL, 35 mmol) and 7-bromo-2-methylbenzofuran 68 (6.7 g, 32 mmol) in DCM (120 ml) at 0°C. The reaction mixture is returned to RT for ~90 min, then poured into ice-cold saturated sodium bicarbonate solution (500 ml). The organic product is separated and the aqueous phase is extracted again with DCM (3x150 ml). The combined organic layers were washed with brine (200 mL), then dried through a phase separator cartridge and concentrated in vacuo. The crude product was purified by recrystallization from EtOAc/hexane (1:1) to give the desired product 69 as a pale yellow solid (4.5 g, 59% yield). R _t 1.48 min (method 2a) m/z 239/241 [M+H]+ (ER+). ¹ H NMR (400 MHz, DMSO-06): δ, ppm 10.20 (s, 1H), 7.98 (dd, J=7.7 Hz, 1.1 Hz, 1H), 7.59 (dd, J=7.9 Hz, 1.1 Hz, 1H), 7.29 (t, J=7.8 Hz, 1H), 2.84 (s, 3H).

Step 4. 1-(7-Bromo-2-methylbenzofuran-3-yl)-N-(4-methoxybenzyl)-N-methylmethanamine (compound 71).

Sodium triacetoxyborohydride (4.4 g, 21 mmol) was added in one portion to a stirred solution of 7-bromo-2-methylbenzofuran-3-carbaldehyde 69 (2 g, 8.37 mmol) and 1-(4-methoxyphenyl)-Nmethylmethanamine 70 ( 1.3 ml, 8.37 mmol) in DCE (40 ml) at RT. The reaction mixture was stirred for ~16 h, then the solvent was concentrated in vacuo and the resulting residue was taken up in NaHCO ₃ (100 ml, sat. aq.). The aqueous product was extracted with DCM (3x100 ml) and the combined organic layers were washed with brine (100 ml), then dried through a phase separator cartridge. The crude product was purified by column chromatography (0-5% MeOH/DCM) to give the desired product 71 as a colorless oil (3.11 g, 92%). R _t 0.98 min (method 2a) m/z 374/376 [M+H]+ (ER+), ¹ H NMR (400 MHz, DMSO-06): δ, ppm 7.59 (dd, J=7.7 Hz, 1.0 Hz, 1H), 7.44 (dd, J=7.8 Hz, 1.0 Hz, 1H), 7.25-7.19 (m, 2H), 7 .16 (t, J=7.8 Hz, 1H), 6.92-6.84 (m, 2H), 3.73 (s, 3H), 3.52 (s, 2H), 3.44 ( s, 2H), 2.45 (s, 3H), 2.04 (s, 3H).

- 64 046887

Step 5. No. (Diphenylmethylene)-3-(((4-methoxybenzyl)(methyl)amino)methyl)-2-methylbenzofuran-7amine (compound 73).

The 20 ml tube is evacuated and backfilled with N2 three times, then 1-(7-bromo-2methylbenzofuran-3-yl)-N-(4-methoxybenzyl)-N-methylmethanamine 71 (0.3 g, 0.80 mmol) is added ), tris(dibenzylideneacetone)dipalladium(0) (75 mg, 0.08 mmol), cesium carbonate (0.52 g, 1.60 mmol) and Xantphos (70 mg, 0.12 mmol) and again evacuated and backfilled N2 three times. Toluene (5 mL) and benzophenonymine 72 (0.14 mL, 0.80 mmol) were then added and the reaction mixture was heated to 100°C and stirred for ~16 h. The reaction mixture was cooled to rt, filtered through Celite® and cake wash with DCM (100 ml). The filtrate was concentrated in vacuo and purified by column chromatography (0-50% EtOAc/isohexane) to give the desired product 73 as a yellow oil (0.36 g, 70% yield). R _t 1.97 min (method 1a); m/z 475 [M+H]+ (ER ⁺ ), ¹ H NMR (500 MHz, DMSO-06): δ, ppm 7.74-7.69 (m, 2H), 7.60-7 .53 (m, 1H), 7.53-7.48 (m, 2H), 7.25-7.21 (m, 3H), 7.21-7.17 (m, 2H), 7.14 -7.07 (m, 3H), 6.94 (t, J=7.7 Hz, 1H), 6.91-6.85 (m, 2H), 6.48 (dd, J=7.6 Hz, 1.1 Hz, 1H), 3.73 (s, 3H), 3.43 (s, 2H), 3.38 (s, 2H), 2.32 (s, 3H), 1.99 ( s, 3H).

Step 6. 2-Methyl-3-((methylamino)methyl)benzofuran-7-amine (compound 74).

1-Chloroethyl chloroformate (0.12 mL, 1.14 mmol) was added dropwise to a stirred solution of Na(diphenylmethylene)-3-(((4-methoxybenzyl)(methyl)amino)methyl)-2-methylbenzofuran-7-amine 73 (0.36 g, 0.76 mmol) in DCM (5 ml) at 0°C under N2. The reaction mixture is warmed to room temperature and stirred for ~16 h. The reaction mixture is concentrated in vacuo, then placed in MeOH (15 ml) and refluxed for 1 h. The reaction mixture is cooled to RT, then concentrated in vacuo and purified by column chromatography (0-10% MeOH (0.7 M NI 1 ₃ )/DC\1) to give the desired product 74 as a yellow oil (78 mg, 46% yield). R _t 0.77 min (method 1b); m/z 191 [M+H] ⁺ (ER ⁺ ), 1H NMR (500 MHz, DMSO-06): δ, ppm 6.86-6.83 (m, 1H), 6.78 (dd, J=7.7 Hz, 1.2 Hz, 1H), 6.46 (dd, J=7.6, 1.2 Hz, 1H), 5.09 (s, 2H), 3.65 (s, 2H), 2.39 (s, 3H), 2.25 (s, 3H). The amine proton has not been determined. /

6 75

Step 7. (E)-N-((7-Amino-2-methylbenzofuran-3-yl)methyl)-N-methyl-3-(4-oxo-2,3,4,5-tetrahydro1H-pyrido[2 ,3-b][1,4]diazepin-8-yl)acrylamide (compound 75).

DIPEA (0.2 ml, 1.15 mmol) is added dropwise to a stirred solution of 2-methyl-3((methylamino)methyl)benzofuran-7-amine 74 (78 mg, 0.41 mmol) and trifluoroacetate (E)- 3-(4-oxo-2,3,4,5tetrahydro-1H-pyrido[2,3-b][1,4]diazepin-8-yl)acrylic acid 6 (50 mg, 0.14 mmol) in DMF (2 ml) at CT. The reaction mixture was stirred for 10 minutes, then GATU (70 mg, 0.18 mmol) was added and the reaction mixture was stirred for 2 hours. The reaction mixture was diluted with water (10 ml) and the precipitate was collected by filtration. The crude product was purified by column chromatography (0-4% MeOH/DCM) to give the desired product 75 as a pale yellow solid (18 mg, 31% yield). R _t 1.30 min (method 1a); m/z 406 [M+H] ⁺ (ER ⁺ ), 1H NMR (500 MHz, DMSO-06, 363 K): δ, ppm 9.13 (s, 1H), 7.97 (d, J =1.9 Hz, 1H), 7.44 (d, J=15.4 Hz, 1H), 7.37 (d, J=2.0 Hz, 1H), 7.08 (d, J=15 .4 Hz, 1H), 6.87 (t, J=7.7 Hz, 1H), 6.74 (d, J=7.7 Hz, 1H), 6.53 (dd, J=7.7 , 1.1 Hz, 1H), 5.79 (s, 1H), 4.86 (s, 2H), 4.71 (s, 2H), 3.46-3.40 (m, 2H), 2 .98 (s, 3H), 2.68-2.61 (m, 2H), 2.46 (s, 3H).

- 65 046887

Example 12. Synthesis of (E)-U-((7-(4-cyanophenoxy)-2-methylbenzofuran-3-yl)methyl)-U-methyl-3-(4-oxo2,3,4,5-tetrahydro- 1H-pyrido[2,3-b][1,4]diazepin-8-yl)acrylamide (compound 80). General synthesis scheme

Reaction conditions: a) Pd-175, tBuBrettPhos, KOH, H ₂ O, dioxane,

80°C; b) 4-fluorobenzonitrile 77,

K2CO3, NMP, 100°C; c) 1-chloroethyl chloroformate, DCM, 0°C to RT; d) GATU, DIPEA, DMF.

Step 1. 3-(((4-Methoxybenzyl)(methyl)amino)methyl)-2-methylbenzofuran-7-ol (compound 76).

A 100 mL round bottom flask was charged with 1-(7-bromo-2-methylbenzofuran-3-yl)-Y-(4-methoxybenzyl)N-methylmethanamine 71 (2.7 g, 7.21 mmol), Pd-175 (Johnson Matthey) (0.11 g, 0.14 mmol), tBuBrettPhos (0.07 g, 0.14 mmol) and potassium hydroxide (1.21 g, 21.6 mmol). It is then evacuated and refilled with nitrogen three times and then degassed 1,4-dioxane (15 ml) and degassed water (2.6 ml) are added. The resulting mixture is heated to 80°C and stirred for 2 hours. The mixture is cooled to RT and diluted with EtOAc (20 ml), then acidified with 1 M HCl (20 ml) and stirred for 5 minutes until all the solid has dissolved. The reaction mixture is neutralized with NaHCO ₃ (25 ml, sat. aq.) and the phases are separated. The aqueous phase is extracted with EtOAc (2x25 ml) and the combined organic phases are washed with brine (25 ml), dried over MgSO ₄ , filtered and concentrated in vacuo. The crude product was purified by chromatography (0-5% MeOH/DCM) to give the title compound 76 as a beige solid (1.80 g, 80%). R _t 1.12 min (method 1a) m/z 312 [M+H]+ (ER+).

Step 2. 4-((3-(((4-Methoxybenzyl)(methyl)amino)methyl)-2-methylbenzofuran-7-yl)oxy)benzonitrile (compound 78).

Mixture of 3-(((4-methoxybenzyl)(methyl)amino)methyl)-2-methylbenzofuran-7-ol 76 (150 mg,

0.48 mmol), 4-fluorobenzonitrile 77 (70 mg, 0.58 mmol) and potassium carbonate (133 mg, 0.96 mmol) in the solvent Namethyl-2-pyrrolidone (6 ml) heated at 100°C for 12 hours. The reaction mixture is cooled to RT and the mixture is poured into water (20 ml). The aqueous phase is extracted with ethyl acetate (2x30 ml) and the combined organic layers are washed with brine (30 ml), dried over Na ₂ SO ₄ and concentrated in vacuo. The crude product was purified by chromatography (1% MeOH/DCM) to give the title compound 78 as a colorless oil (170 mg, 86%). R _t 1.65 min (method 1a) m/z 413 [M+H]+ (ER+).

- 66 046887

Step 3. 4-((2-methyl-3-((methylamino)methyl)benzofuran-7-yl)oxy)benzonitrile hydrochloride (compound 79).

To an ice-cold solution of 4-((3-(((4-methoxybenzyl)(methyl)amino)methyl)-2-methylbenzofuran-7yl)oxy)benzonitrile 78 (185 mg, 0.45 mmol) in DCM (2 ml) 1-chloroethyl carbonochloridate (0.10 ml, 0.90 mmol) at 0°C. The reaction mixture is warmed to RT and stirred for 1 hour and then concentrated in vacuo. The residue was dissolved in methanol (5 ml) and heated to reflux for 1 hour. The reaction mixture was evaporated to dryness and triturated with TBME (10 ml) to obtain the title compound 79 as a yellow solid (185 mg, 47% ). R _t 1.31 min (method 1a) m/z 293 [M+H]+ (ER ⁺ ).

Step 4. (E)-N-((7-(4-Cyanophenoxy)-2-methylbenzofuran-3-yl)methyl)-N-methyl-3-(4-oxo-2,3,4,5tetrahydro-1 H-pyrido[2,3-b][1,4]diazepin-8-yl)acrylamide (compound 80).

A solution of 4-((2-methyl-3-((methylamino)methyl)benzofuran-7-yl)oxy)benzonitrile hydrochloride 79 (69 mg, 0.20 mmol) in DMF (4 ml) was treated with trifluoroacetate (E)-3 -(4-oxo-2,3,4,5-tetrahydro-1Hpyrido[2,3-b][1,4]diazepin-8-yl)acrylic acid 6 (80 mg, 0.23 mmol), Huenig's base (180 µl, 1.05 mmol) and HATU (96 mg, 0.25 mmol) and then stirred at RT for 1 hour. The reaction mixture was quenched with water (10 ml) and the solid was collected by filtration and dried. The crude product was purified by column chromatography (0-10% MeOH/DCM) to give the title compound 80 as a pale yellow solid (23 mg, 21% yield). R _t 2.01 min (method 1a) m/z 508 [M+H]+ (ER+), ¹ H NMR (500 MHz, DMSO-dfo 363 K): δ, ppm 9.15 (s, 1h), 7.99 (d, J=1.9 Hz, 1H), 7.83-7.77 (m, 2H), 7.50-7.44 (m, 2H), 7.38 (d , J=2.0 Hz, 1H), 7.25 (t, J=7.9 Hz, 1H), 7.15-7.06 (m, 3H), 7.06-7.01 (m, 1H), 5.81 (s, 1H), 4.79 (s, 2H), 3.47-3.40 (m, 2H), 3.04 (s, 3H), 2.69-2.62 (m, 2H), 2.46 (s, 3H).

Example 13. Synthesis of (E)-3-(2-(hydroxymethyl)-2-methyl-3-oxo-1,2,3,4-tetrahydropyrido[2,3b]pyrazin-7-yl)-N-methyl- N-((3-methylbenzofuran-2-yl)methyl)acrylamide (compound 84). General synthesis scheme.

Reaction conditions: a) K2CO3, THF; b) NH4C1, Fe, EtOH/water; c) Pd(OAc)2, tri-o-tolylphosphane, DIPEA in DMF/ACN.

- 67 046887

Step 1. 2-((5-Bromo-2-nitropyridin-3-yl)amino)-3-hydroxy-2-methylpropanoic acid (compound 82).

5-Bromo-3-fluoro-2-nitropyridine 11 (800 mg, 3.62 mmol, 1.0 equiv.) and 2-amino-3-hydroxy-2methylpropanoic acid 81 (Sigma-Aldrich) (736 mg, 5. 07 mmol, 1.4 eq.) dissolved in THF (16.0 ml), then K ₂ CO ₃ (999 mg) was added. The resulting reaction mixture is heated to 90°C overnight in a sealed tube. UPLC-MS analysis showed almost complete conversion of IM with the formation of a good product (R _t = 0.94 min; ER ⁺ = 322.0). The solvent is removed under vacuum. The residue is dissolved in a mixture of water and EtOAc. The aqueous layer is acidified with 1 M HCl to pH 1-2. The phases are separated and aq. the phase is re-extracted with EtOAc. The combined organic phases are washed with brine, dried over MgSO ₄ and concentrated to dryness. The crude product (800 mg) was purified by an Isolera system using AcOEt/MeOH (100% to 80/20 v/v) to give the title compound 82 as an orange solid with 60% purity (320 mg, 17% yield ). A second batch with 80% purity is also isolated (40 mg, 3.45% yield). R _t 0.94 min (method 1c). UPLC-MS m/z: 322.0 [M+H] ⁺ (ER ⁺ ).

Step 2. 7-Bromo-2-(hydroxymethyl)-2-methyl-1,4-dihydropyrido[2,3-b]pyrazin-3(2H)-one (compound 83).

To a stirred solution of 2-((5-bromo-2-nitropyridin-3-yl)amino)-3-hydroxy-2methylpropanoic acid 82 (40 mg, 0.124 mmol, 1.0 eq.), NH4Cl (13.4 mg, 0.25 mmol, 2.0 eq.) and iron (14 mg, 0.25 mmol, 2.0 eq.) were suspended in EtOH/Water (1:1 v/v, 2 ml). The resulting suspension is heated to 80°C for 1.5 hours. UPLC analysis showed the formation of a product (Rt = 0.89 min; ER ⁺ = 272.1) with complete conversion of the starting material. The reaction mixture is diluted with EtOAc/MeOH, filtered through a 0.22 µM filter and the filter washed with acetonitrile. The resulting solution is evaporated to dryness, then diluted with AcOEt, washed with water, dried (MgSO4) and concentrated. Crude product 83 (37 mg, 0.136 mmol) was isolated as a red solid with a purity of 60%. The resulting product is used in the next step without purification. R _t 0.89 min (method 1c). LCMS: m/z: 272.1 [M+H] ⁺ (ER ⁺ ) ¹ H NMR (400 MHz, DMSO^): δ, ppm 10.80 (s, 1H), 7.52 (s, 1H), 7.07 (s, 1H), 6.47 (s, 1H), 5.12 (t, J=5.6 Hz, 1H), 3.58 (dd, J=6.1 Hz, 10.4 Hz, 1H), 3.29 (s, 1H), 1.25 (s, 3H).

Vg

n < ^he

Ν ^Χ Ό N

Pd(OAc) ₂ tri-o-tolylphosphane

DIPEA DMF/acetanitrile

83 84

Step 3. (E)-3-(2-(Hydroxymethyl)-2-methyl-3-oxo-1,2,3,4-tetrahydropyrido[2,3-b]pyrazin-7-yl)Nmethyl-N ((3-methylbenzofuran-2-yl)methyl)acrylamide (compound 84).

7-bromo-2-(hydroxymethyl)-2-methyl-1,4dihydropyrido[2,3-b]pyrazin-3(2H)-one 83 (37 mg, 0.13 mmol) is loaded into an oven-dried test tube, and anhydrous DMF (0.9 ml) and ACN (0.1 ml) and the reaction mixture was degassed with nitrogen for 5 min. Namethyl-Na((2methylbenzofuran-3-yl)methyl)acrylamide 9 (47 mg, 0.204 mmol) and DIPEA (93 µl) were then added and the mixture was degassed a second time for 10 min. Finally, Pd(OAc) ₂ (0.3 mg) and tri-o-tolylphosphane (0.83 mg) are added in one portion and nitrogen is bubbled through the mixture for 1 minute and the solution is heated at 100°C overnight. The reaction mixture is cooled to RT, filtered on a 0.22 µM filter, washed with MeOH/AcOEt and concentrated under high vacuum. The crude brown oil is purified with Isolera (nheptane/AcOEt 7/3 v/v ^ 100% AcOEt ^ AcOEt/MeOH 8/2 v/v) and then re-purified 2 more times with DCM/MeOH (100% to 95/ 5 v/v). The target molecule 84 was isolated as a beige solid with 94% purity (4.9 mg, 8.6% yield). R _t 1.47 min (method 1c). LCMS: m/z: 421.3 [M+H] ⁺ (ER ⁺ ). 1H NMR (400 MHz, DMSO^): δ, ppm 10.85 (s, 1H), 7.80 (s, 1H), 7.58 (d, J=7.4 Hz, 1H), 7 .51 (d, J=7.4 Hz, 1H), 7.43 (d, J=16.1 Hz, 1H), 7.32-7.22 (m, 4H), 6.99 (d, J=14.4 Hz, 1H), 6.20 (d, J=13.1 Hz, 1H), 5.06 (t, J=5.5 Hz, 1H), 4.95 (s, 2H) , 4.80 (s, 2H), 3.62-3.53 (m, 1H), 2.96 (s, 1H), 2.28 (s, 3H), 1.25 (s, 6H) ( rotamers).

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Example 14. Synthesis of 8-((E)-3-oxo-3-((3r6,6L)-5-(4-(pyridin-4-yloxy)phenyl)-3.3g,4.6gtetrahydrocyclopenta[c]pyrrole -2(1H)-yl)prop-1-en-1-yl)-1,2,3,5-tetrahydro-4H-pyrido[2,3-b][1,4]diazepin-4-one (compound 90).

General synthesis scheme.

Reaction conditions: a) B ₂ (pin) ₂ 85, KOAc, PdCl ₂ (dppf), 1,4-dioxane, 100°C; b) 4-(4bromophenoxy)pyridine 87, K ₂ CO ₃ , H ₂ O, PdCl ₂ (dppf), 1,4-dioxane, 80°C; c) TPA, DXM; d) GATU, DIPEA,

DMF.

B ₂ (Pin) ₂ 85, PdCI ₂ (dppf),

KOAc, 1,4-dioxane, 100 °C

Stage 1. tert-Butyl (3gR,6gS)-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-3.3g,4.6gtetrahydrocyclopenta^pyrrole^III carboxylate ( connection 86).

Mixture of tert-butyl 5-(((trifluoromethyl)sulfonyl)oxy)-3.3 g,6.6 g-tetrahydrocyclonent[c]nirrole2(III)-carboxylate 57 (2 g, 5.6 mmol), 4,4,4 ',4',5,5,5',5'-octamethyl-2,2'-bi(1,3,2-dioxaborolane) 85 (2.42 g, 9.5 mmol), potassium acetate (1, 65 g, 16.8 mmol) and the PdCl2(dppf)-CH2Cl2 adduct (0.46 g, 0.6 mmol) in 1,4dioxane (20 ml) were degassed with nitrogen and then heated at 100°C overnight. The mixture is diluted with water (30 ml), extracted with ethyl acetate (2x30 ml). The combined organic layers were washed with brine (30 ml), dried over MgSO ₄ , filtered and evaporated to dryness. The crude product was purified by column chromatography (0-50% EtOAc/isohexane) to give the title compound 86 as a colorless oil (1.97 g, 94%), ¹ H NMR (400 MHz, CDCl ₃ ): δ, h/ ppm 6.35 (kV, J=2.0 Hz, 1H), 3.66-3.61 (m, 1H), 3.54-3.44 (m, 2H), 3.39 (ddt, J =8.0 Hz, 5.7 Hz, 2.5 Hz, 1H), 2.97 (dd, J=10.9 Hz, 7.2 Hz, 1H), 2.89 (dd, J=9, 0 Hz, 7.3 Hz, 1.6 Hz, 1H), 2.65 (ddt, J=16.5 Hz, 7.2 Hz, 2.6 Hz, 1H), 2.38 (dt, J= 16.5 Hz, 1.8 Hz, 1H), 1.46 (s, 9H), 1.29 (s, 12H), 1.28 (s, 3H).

Step 2. tert-Butyl (3gR,6gS)-5-(4-(pyridin-4-yloxy)phenyl)-3.3g,4.6g-tetrahydrocyclopenta^pyrrole^III^carboxylate (compound 88).

Mixture of 4-(4-bromophenoxy)pyridine 87 (enamine) (149 mg, 0.60 mmol), tert-butyl 5-(4,4,5,5tetramethyl-1,3,2-dioxaborolan-2-yl)- 3.3g,6.6g-tetrahydrocyclopenta[c]pyrrole-2(1H)-carboxylate 86 (200 mg, 0.6 mmol), K2CO3 (247 mg, 1.8 mmol) and PdCl2(dppf)-CH2Cl2 adduct ( 49 mg, 0.06 mmol) in 1,4dioxane (3 ml) is degassed with nitrogen and then heated at 80°C for 5 hours. The mixture is diluted with water (30 ml), extracted with ethyl acetate (2x30 ml). The combined organic layers were washed with brine (30 ml), dried over MgSO ₄ , filtered and evaporated to dryness. The crude product was purified by chromatography (0-5% MeOH/DCM) to give the title compound 88 as a brown oil (193 mg, 77%). Rt 2.49 min (method 1b); m/z 379 [M+H]+ (ER ⁺ ), 1H NMR (400 MHz, CDCl ₃ ): δ, ppm 8.47 (d, J=5.6 Hz, 2H), 7.53- 7.46 (m, 2H), 7.09-7.04 (m, 2H), 6.90-6.81 (m, 2H), 6.03 (s, 1H), 3.71-3, 45 (m, 4H), 3.14-2.93 (m, 3H), 2.60 (s, 2H), 1.46 (s, 9H).

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Step 3. (3gR,6gS)-5-(4-(Pyridin-4-yloxy)phenyl)-1,2,3,3g,4,6g-hexahydrocyclopenta[c]pyrrole (compound 89).

Trifluoroacetic acid (1.5 ml) is added dropwise to the stirred solution of tert-butyl 5-(4-(pyridin-4-yloxy)phenyl)-3.3 g,6.6 g-tetrahydrocyclopenta[c]pyrrole-2(1H) -carboxylate 88 (193 mg, 0.5 mmol) in DCM (1.5 ml) at RT. The reaction mixture was stirred for 2 hours, then the solvent was concentrated in vacuo and the resulting residue was taken up in methanol (30 ml) and applied to an SCC column. The column is washed with methanol (50 ml) and the product is eluted with 10% methanolic ammonia. The mixture was then evaporated to give the title compound 89 as a yellow oil (115 mg, 73%). Rt 1.36 min (method 1b); m/z 279 [M+H]+ (ER+), 1H NMR (400 MHz, CDCl ₃ ): δ, ppm 8.48-8.41 (m, 2H), 7.44 (d, J =8.3 Hz, 2H), 7.02 (d, J=8.3 Hz, 2H), 6.87-6.76 (m, 2H), 5.95-5.94 (m, 1H) , 3.78 (s, 2H), 3.51 (d, J=7.4 Hz, 1H), 3.10-2.80 (m, 5H), 2.51 (dd, J=15.6 , 2.6 Hz, 1H).

Step 4. 8-((E)-3-Oxo-3-((3r6,6G)-5-(4-(pyridin-4-yloxy)phenyl)-3.3g,4.6g-tetrahydrocyclopenta[s] pyrrol-2(1H)-yl)prop-1-en-1-yl)-1,2,3,5-tetrahydro-4H-pyrido[2,3-b][1,4]diazepin-4-one (compound 90).

Suspension of (3gS,6gR)-5-(4-(nirigin-4-yloxy)phenyl)-1,2,3,3g,4,6g-hexahydrocyclopenta[c]nirrole 89 (115 mg, 0.4 mmol), trifluoroacetate (E)-3-(4-oxo-2,3,4,5-tetrahydro-1H-nirigo[2,3-b][1,4]diazenin8-yl)acrylic acid 6 (136 mg, 0. 4 mmol), X-ethyl-X-isopropylpropan-2-amine (0.28 ml, 1.6 mmol) in DMF (2 ml) was stirred for 10 min. GATU (157 mg, 0.4 mmol) was added in one portion and the reaction mixture was stirred at RT for 1 hour. The mixture was diluted with water (10 ml) and the precipitate was collected by filtration. The crude product was purified by column chromatography (0-5% MeOH/DCM) to give the title compound 90 as a yellow solid (92 mg, 45% yield). Rt 1.74 min (method 1b); m/z 494 [M+H]+ (ER+), ¹ H NMR (400 MHz, DMSO^): δ, ppm 9.73 (d, J=6.6 Hz, 1H), 8.47 -8.43 (m, 2H), 7.98 (dd, J=8.2 Hz, 2.0 Hz, 1H), 7.61-7.56 (m, 2H), 7.38-7, 30 (m, 2H), 7.18-7.13 (m, 2H), 6.93-6.83 (m, 3H), 6.22 (d, J=3.6 Hz, 1H), 6 .05-5.98 (m, 1H), 4.00 (t, J=9.9 Hz, 1H), 3.88-3.78 (m, 1H), 3.70 (m, 1H), 3.65-3.52 (m, 1H), 3.45-3.39 (m, 2H), 3.21-3.13 (m, 1H), 3.04-2.88 (m, 2H ), 2.68-2.57 (m, 3H).

Example 15. Synthesis of 8-((E)-3-oxo-3-((3gS,6gR)-5-(4-(thiazol-5-yloxy)phenyl)-3.3g,4.6g-tetrahydrocyclopenta[c ]pyrrol-2(1H)-yl)nrop-1-en-1-yl)-1,2,3,5-tetrahydro-4H-nirido[2,3-b][1,4]diazepine-4- she (compound 96).

General synthesis scheme.

Reaction conditions: a) 5-bromothiazole 92, Cs ₂ CO ₃ , CuI, DMF, 110°C; b) Pd(dppf)Cl ₂ , K ₂ CO ₃ , 1,4-dioxane, 90°C; c) TPA, DXM: d) GATU, DIPEA, DMF.

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YY ^CS2C ° ^3, nsg''^ S'^'Br Cui, DMF, 110 C

92 93

Step 1. 5-(4-Bromophenoxy)thiazole (compound 93).

To a stirred solution of 5-bromothiazole 92 (Fluorochem) (0.2 g, 1.2 mmol) in DMF (6 ml) add 4-bromophenol 91 (0.25 g, 1.5 mmol), copper iodide ([) ( 0.23 g, 1.2 mmol), then cesium carbonate (1.19 g, 3.7 mmol). The reaction mixture is stirred at 110°C for 5 hours and cooled to room temperature. The reaction mixture is partitioned between water (5 ml) and EtOAc (5 ml). The phases are separated and the aqueous phase is extracted with EtOAc (2x10 ml). The combined organic phases were washed with brine (15 mL), dried over MgSO ₄ and concentrated in vacuo to give the title compound 93 as a white solid (0.26 g, 84%). R _t 2.19 min (method 1a) m/z 256/258 [M+H]+ (ER+).

⁹³ 86 94

Step 2. tert-Butyl (3rS,6rR)-5-(4-(thiazol-5-yloxy)phenyl)-3,3r,4,6r-tetrahydrocyclopenta^pyrrole^III carboxylate (compound 94).

Mixture of 5-(4-bromophenoxy)thiazole 93 (0.17 g, 0.7 mmol), tert-butyl 5-(4,4,5,5-tetramethyl-1,3,2dioxaborolan-2-yl)-3 ,3r,6,6r-tetrahydrocyclopenta[c]pyrrole-2(1H)-carboxylate 86 (0.22 g, 0.7 mmol), K ₂ CO ₃ (0.28 g, 2.0 mmol) and PdCl adduct ₂ (dppf)-CH ₂ Cl ₂ (0.05 g, 0.07 mmol) in 1,4-dioxane (4 ml) was degassed with nitrogen and then stirred at 90°C overnight. The reaction mixture is cooled to room temperature, diluted with water (10 ml) and extracted with ethyl acetate (2x10 ml). The combined organic layers were washed with brine (20 ml), dried (MgSO ₄ ), filtered and concentrated in vacuo. The crude product was purified by column chromatography (0-50% EtOAc/isohexane) to give the title compound 94 as a thick, colorless oil (0.06 g, 23% yield). Rt 2.68 min (method 1a) m/z 329 [M+H-tBu]+ (ER+), ¹ H NMR (400 MHz, DMSO-06): δ, ppm 8.77 (s, 1H ), 7.63 (s, 1H), 7.51 (d, J=8.7 Hz, 2H), 7.12 (d, J=8.7 Hz, 2H), 6.13 (s, 1H ), 3.62-3.53 (m, 1H), 3.49-3.35 (m, 3H), 3.11-2.81 (m, 4H), 1.37 (s, 9H).

95

Step 3. 5-(4-((3rS,6rR)-1,2,3,3r,4,6r-Hexahydrocyclopenta[c]pyrrol-5-yl)phenoxy)thiazole (compound 95).

To a stirred solution of (3r6,6G)-tert-butyl 5-(4-(thiazol-5-yloxy)phenyl)-3,3r,6,6-tetrahydrocyclopenta^pyrrolXIIIYcarboxylate 94 (0.06 g, 0.2 mmol) in DCM (1 ml) add TFA (1 ml). The reaction mixture is stirred at room temperature for 2 hours. The solvent is removed in vacuo. The resulting oil was taken up in MeOH (5 mL) and applied to an SCC column. The column was washed with methanol (5 mL) and the product was eluted with 10% methanolic ammonia (5 mL) to give the title compound 95 as a colorless crystalline solid (34 mg, 74%). The crude product is used in the next step without purification. R _t 1.17 min (method 1a) m/z 285 [M+H]+ (ER ⁺ ).

96

Step 4. 8-((E)-3-Oxo-3-((3aS,6aR)-5-(4-(thiazol-5-yloxy)phenyl)-3,3a,4,6a-tetrahydrocyclopenta[c] pyrrol-2(1H)-yl)prop-1-en-1-yl)-1,2,3,5-tetrahydro-4H-pyrido[2,3-b][1,4]diazepin-4-one (compound 96).

To a stirred solution of 5-(4-((3rS,6rR)-1,2,3,3r,4,6r-hexahydrocyclopenta[c]pyrrol-5yl)phenoxy)thiazole 95 (34 mg, 0.1 mmol) in DMF (2 ml) add trifluoroacetate (E)-3-(4-oxo-2,3,4,5

- 71 046887 tetrahydro-1H-pyrido[2,3-b][1,4]diazepin-8-yl)acrylic acid 6 (31 mg, 0.13 mmol) and Ν-θτήλ-Νisopropylpropan-2-amine (0 .1 ml, 0.6 mmol). The reaction mixture was stirred at RT for 5 min and GATU (55 mg, 0.1 mmol) was added. The resulting mixture was stirred for 3 hours. Water (1 ml) was added and the reaction mixture was treated with ultrasound. The solid is filtered and washed with H2O (2x2 ml). The filtrate is discarded. The crude solid was purified by column chromatography (0-10% MeOH/DCM) to give the title compound 96 as a yellow solid (25 mg, 40%). Rt 1.85 min (method 1a) m/z 500 [M+H]+ (ER+), ¹ H NMR (400 MHz, DMSO-dg): δ, ppm 9.74 (d, J=7 .3 Hz, 1H), 8.76 (d, J=1.3 Hz, 1H), 7.98 (dd, J=9.1 Hz, 1.9 Hz, 1H), 7.63 (t, J=1.2 Hz, 1H), 7.53 (d, J=8.7 Hz, 2H), 7.39-7.28 (m, 2H), 7.20-7.09 (m, 2H ), 6.86 (dd, J=15.5 Hz, 9.6 Hz, 1H), 6.17 (s, 1H), 6.02 (d, J=14.0 Hz, 1H), 4, 02-3.78 (m, 2H), 3.68 (d, J=12.1 Hz, 1H), 3.64-3.49 (m, 2H), 3.45-3.35 (m, 2H), 3.19-3.08 (m, 1H), 3.03-2.87 (m, 2H), 2.64-2.58 (m, 2H).

Example 16. Synthesis of (E)-N-methyl-N-((2-methyl-7-(thiazol-5-yloxy)benzofuran-3-yl)methyl)-3-(4-oxo2,3,4,5 -tetrahydro-1H-pyrido[2,3-b][1,4]diazepin-8-yl)acrylamide (compound 101).

General synthesis scheme.

Reaction conditions: a) i) NH2Me, TABN, EtOH; ii) Boc ₂ O, DMAP, DCM; b) Pd-175, tBuBrettPhos, KOH, 1,4-dioxane, H ₂ O; c) 5-bromothiazole 92, CuI, N,N-dimethylglycine, Cs ₂ CO ₃ , DMF; d) TPA, DXM; e) GATU, DIPEA, DMF.

Step 1. tert-Butyl ((7-bromo-2-methylbenzofuran-3-yl)methyl)(methyl)carbamate (compound 97).

Methamine (33% in EtOH, 0.5 mL, 4.4 mmol) is added dropwise to a stirred solution of 7bromo-2-methylbenzofuran-3-carbaldehyde 69 (2.8 g, 3.7 mmol) and sodium triacetoxyborohydride (TABN) (1.9 g, 9.2 mmol) in EtOH (100 ml) at 0°C. The reaction mixture is returned to RT and stirred for 5 hours. The reaction mixture is then concentrated in vacuo and the resulting residue is taken up in EtOAc (100 ml) and NaHCO ₃ (aq 100 ml). The organic product is separated and the aqueous phase is extracted with EtOAc (2x100 ml). The combined organic layers were washed with brine (100 mL), dried with MgSO ₄ and concentrated in vacuo. The resulting residue was dissolved in DCM (80 ml), then DMAP (0.6 g, 4.6 mmol) and di-tert-butyl dicarbonate (1.6 g, 7.3 mmol) were added and the reaction mixture was stirred at RT for 16 hours. The crude reaction mixture was concentrated in vacuo and purified by column chromatography (0-50% EtOAc/isohexane) to give the title compound 97 as a yellow oil that crystallized on standing (1.0 g, 69% yield). R _t 1.86 min (method 1a) m/z 376/378 [M+Na]+ (ER ⁺ ), ¹ H NMR (400 MHz, DMSO-de): δ, ppm 7.55 (shs, 1H), 7.46 (d, J=7.8 Hz, 1H), 7.17 (shs, 1H), 4.48 (s, 2H), 2.67 (s, 3H), 2.51 ( s, 3H), 1.44 (s, 9H).

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Step 2. tert-Butyl ((7-hydroxy-2-methylbenzofuran-3-yl)methyl)(methyl)carbamate (compound 98).

1,4-Dioxane (5 ml) and degassed H ₂ O (1.5 ml) are added to a mixture of tert-butyl ((7-bromo-2methylbenzofuran-3-yl)methyl)(methyl)carbamate 97 (1.0 g , 2.9 mmol), Pd-175 (0.06 g, 0.07 mmol), tBuBrettPhos (0.03 g, 0.07 mmol) and KOH (0.5 g, 8.8 mmol) under nitrogen , and the reaction mixture was stirred at 80°C for 2 hours. The reaction mixture was cooled to RT, then poured into water (50 ml). The aqueous mixture is adjusted to pH 6 using 1 M HCl and extracted with EtOAc (3x50 ml). The combined organic extracts were washed with brine (50 ml), dried using MgSO ₄ and concentrated in vacuo. The crude product was purified by column chromatography (0-50% EtOAc/isohexane) to give the title compound 98 as a pale brown solid (0.58 g, 67% yield). Rt 2.18 min (method 1a) m/z 314 [M+Na]+ (ER+), ¹ H NMR (400 MHz, DMSO-06): δ, ppm 9.85 (s, 1H), 6.96 (shs, 2H), 6.76-6.56 (m, 1H), 4.44 (s, 2H), 2.65 (s, 3H), 2.44 (s, 3H), 1 .44 (s, 9H).

Step 3. tert-Butyl methyl ((2-methyl-7-(thiazol-5-yloxy)benzofuran-3-yl)methyl)carbamate (compound 99).

DMF (5 ml) is added to a mixture of Cs ₂ CO ₃ (0.9 g, 2.8 mmol), 5-bromothiazole 92 (0.17 g, 1.03 mmol), tert-butyl ((7-hydroxy-2 -methylbenzofuran-3-yl)methyl)(methyl)carbamate 98 (0.2 g, 0.686 mmol), 2(dimethylamino)acetic acid (0.07 g, 0.69 mmol) and CuI (0.13 g, 0 .69 mmol) under N2 atmosphere, and the reaction mixture was heated to 110°C. The reaction mixture was stirred at this temperature for 24 hours, then an additional equivalent of 5-bromothiazole (0.17 g, 1.03 mmol) was added and the reaction mixture was stirred for a further 16 hours at the same temperature. The reaction mixture is cooled to RT, then stirred over the weekend. The crude mixture was dry loaded into Celite® and purified by column chromatography (0-100% EtOAc/isohexane) to give the title compound 99 as a yellow oil (54 mg, 20% yield). Rt 2.51 min (method 1a) m/z 375 [M+H]+ (ER ⁺ ).

Step 4. Nmethyl-1-(2-methyl-7-(thiazol-5-yloxy)benzofuran-3-yl)methanamine trifluoroacetate (compound 100).

Trifluoroacetic acid (2.5 ml) is added dropwise to a stirred solution of tert-butyl methyl ((2-methyl-7-(thiazol-5-yloxy)benzofuran-3-yl)methyl)carbamate 99 (0.05 g, 0 .13 mmol) in DCM (2.5 ml) at RT. The reaction mixture was stirred for 30 minutes, then concentrated in vacuo to give 100 as a yellow oil, which was used in the next step without further purification. Rt 1.39 min (method 1a) m/z 275 [M+H] ⁺ (ER ⁺ ).

Step 5. (E)-N-Methyl-N-((2-methyl-7-(thiazol-5-yloxy)benzofuran-3-yl)methyl)-3-(4-oxo-2,3,4, 5tetrahydro-1H-pyrido[2,3-b][1,4]diazepin-8-yl)acrylamide (compound 101).

DIPEA (0.5 mL, 2.86 mmol) was added dropwise to a stirred solution of Nmethyl-1-(2-methyl-7-(thiazol-5-yloxy)benzofuran-3-yl)methanamine 100 trifluoroacetate (0.05 g , 0.13 mmol) and trifluoroacetate (E)-3-(4-oxo-2,3,4,5-tetrahydro-1H-pyrido[2,3-b][1,4]diazepin-8-yl) acrylic acid 6 (0.06 g, 0.16 mmol) in DMF (2 ml) at RT. The reaction mixture was stirred for 10 minutes, then GATU (0.08 g, 0.20 mmol) was added and the reaction mixture was stirred for 16 hours. The reaction mixture was diluted with water (10 ml) and the precipitate was collected by filtration. The crude product was purified by column chromatography (0-5% MeOH/DCM) to give the title compound 101 as a tablet

- 73 046887 yellow solid (57 mg, 73% yield). R _t 1.78 min (method 1a) m/z 490 [M+H]+ (ER+), ¹ H NMR (400 MHz, DMSO-dQ: δ, ppm 9.14 (s, 1H), 8.66 (s, 1H), 7.98 (d, J=1.9 Hz, 1H), 7.58 (s, 1H), 7.46 (d, J=15.4 Hz, 1H), 7.43-7.35 (m, 2H), 7.19 (t, J=7.9 Hz, 1H), 7.08 (d, J=15.5 Hz, 1H), 7.02 (d , J=8.1 Hz, 1H), 5.79 (t, J=4.2 Hz, 1H), 4.78 (s, 2H), 3.48-3.39 (m, 2H), 3 .02 (s, 3H), 2.67-2.62 (m, 2H), 2.51 (s, 3H).

Example 17. Synthesis of (E)-N-((7-amino-2-methylbenzofuran-3-yl)methyl)-3-(3,3-dimethyl-4-oxo-2,3,4,5tetrahydro-1H- pyrido[2,3-b][1,4]diazepin-8-yl)-N-methylacrylamide (compound 109).

General synthesis scheme.

Reaction conditions: a) K ₂ CO ₃ , THF, reflux; b) Fe, AcOH, EtOH, 90°C; c) LiOH, THF:H ₂ O, RT; d) GATU, DIPEA, DMF, KT; e) tert-butyl acrylate, Pd-116, DIPEA, NBu ₄ Cl, 1,4 dioxane, 90°C; f) TPA, DXM; g) GATU, DIPEA, DMF.

Step 1. Methyl 3-((5-bromo-2-nitropyridin-3-yl)amino)-2,2-dimethylpropanoate (compound 103).

To a solution of 5-bromo-3-fluoro-2-nitropyridine 11 (0.34 g, 1.55 mmol) in THF (5 ml) add methyl 3-amino-2,2-dimethylpropanoate hydrochloride a102 (0.20 g, 1.55 mmol) and K2CO3 (0.43 g, 3.10 mmol). The reaction mixture is stirred at reflux overnight. The reaction mixture is cooled to room temperature, diluted with water (15 ml), and extracted with EtOAc (3x15 ml). The organic layers were combined, dried over MgSO ₄ , filtered, and concentrated in vacuo to give the title compound 103 (0.51 g, 82% yield) as a yellow oil. The crude product is used in the next step without purification. R _t 2.15 min (method 1a); m/z 332/334 [M+H]+ (ER+).

N\/n\/

XX I -—XI I

N NO ₂ N NH ₂

103 104

Step 2. Methyl 3-((2-amino-5-bromopyridin-3-yl)amino)-2,2-dimethylpropanoate (compound 104).

Acetic acid ( 1.80 ml, 30.9 mmol), then iron (0.86 g, 15.5 mmol). The reaction mixture is stirred at 90°C for 2 hours. The resulting mixture is cooled to room temperature and neutralized to pH 8 with solid NaHCO ₃ . Stirring is continued until gas evolution ceases. The reaction mixture is diluted with H2O (5 ml) and EtOAc (5 ml). The aqueous phase is extracted with EtOAc (2x5 ml). The combined organic phases are washed with H2O (10 ml), dried over MgSO ₄ , filtered, and concentrated in vacuo to give the title compound 104 (424 mg, 84% yield) as a brown solid. The crude product is used in the next step without purification. R _t 1.25 min (method 1a); m/z 302/304 [M+H]+ (ER+).

104

105

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Step 3. 3-((2-Amino-5-bromopyridin-3-yl)amino)-2,2-dimethylpropanoic acid (compound 105).

A solution of lithium hydroxide (170 mg, 7.01 mmol) in H ₂ O (1 ml). The reaction mixture was stirred at room temperature for 3 hours. The solvent was removed in vacuo to give the title compound 105 (404 mg, 1.40 mmol, qty) as a brown solid. The crude product is used in the next step without purification. R _t 1.10 min (method 1a); m/z 288/290 [M+H]+ (ER+).

105 106

Step 4. 8-Bromo-3,3-dimethyl-2,3-dihydro-1H-pyrido[2,3-b][1,4]diazepin-4(5H)-one (compound 106).

DIPEA (730 μL, 4.21 mmol), then GATU (800 mg, 2.10 mmol). The reaction mixture is stirred at room temperature for 1 hour and then aq. is added. 1 M HCl (3 ml). The aqueous phase is extracted with DCM (2x5 ml). The combined organic extracts are washed with H2O (3x5 ml), dried over MgSO ₄ , filtered, and concentrated in vacuo. The crude product is purified by chromatography on silica (0-5% MeOH in DCM) to give 8bromo-3,3-dimethyl-2,3-dihydro-1H-pyrido[2,3-b][1,4]diazepine-4 (5H)-one 106 (317 mg, 84% yield) as a white solid. R _t 1.68 min (method 1a); m/z 270/272 [M+H]+ (ER+), ¹ H NMR (400 MHz, CDCl ₃ ): δ, ppm, 7.97 (s, 1H), 7.72 (d, J =2.0 Hz, 1H), 7.04 (d, J=2.0 Hz, 1H), 3.09 (s, 2H), 1.23 (s, 6H).

106 107

Step 5. tert-Butyl (E)-3-(3,3-dimethyl-4-oxo-2,3,4,5-tetrahydro-1H-pyrido[2,3-b][1,4]diazepine8- yl)acrylate (compound 107).

DIPEA (1.3 mL, 7.4 mmol) and tert-butyl acrylate (1.1 mL, 7.5 mmol) were added dropwise to a stirred solution of 8-bromo-3,3-dimethyl-2,3-dihydro- 1H-pyrido[2,3-b][1,4]diazepin-4(5H)-one 106 (1 g, 3.7 mmol), Bu ₄ NCl (0.1 g, 0.4 mmol) and [ P(tBu) ₃ ] ₂ Pd (Pd-116, Johnson Matthey) (0.1 g, 0.2 mmol) in 1,4dioxane (20 ml) under N2. The reaction mixture is heated to 90°C and stirred for 2 hours. The reaction mixture is cooled to RT and concentrated in vacuo. The crude product was purified by column chromatography (0-100% EtOAc/isohexane) to give the title compound 107 as a yellow oil (0.97 g, 80% yield). R _t 2.07 min (method 1a); m/z 318 [M+H]+ (ER+), 1H NMR (400 MHz, DMSO-06): δ, ppm 9.51 (s, 1H), 7.93 (d, J=2, 0 Hz, 1H), 7.43 (d, J=16.0 Hz, 1H), 7.33 (d, J=2.0 Hz, 1H), 6.34 (d, J=16.0 Hz , 1H), 6.21 (s, 1H), 3.05 (d, J=4.4 Hz, 2H), 1.48 (s, 9H), 1.15 (s, 6H).

107 108

Step 6. TPA salt (E)-3-(3,3-dimethyl-4-oxo-2,3,4,5-tetrahydro-1H-pyrido[2,3-b][1,4]diazepin-8yl )acrylic acid (compound 108).

Trifluoroacetic acid (5 ml) is added dropwise to the stirred solution of (E)-tert-butyl 3-(3,3-dimethyl-4-oxo-2,3,4,5-tetrahydro-1H-pyrido[2,3-b] [1,4]diazepin-8-yl)acrylate 107 (0.97 g, 3.1 mmol) in CH ₂ Cl ₂ (10 ml) at RT and the reaction mixture was stirred for 2 hours. The reaction mixture was concentrated in vacuo and the resulting solid was triturated with MTBE (50 mL) then collected by filtration to give the desired product 108 as a yellow solid (0.81 g, 70%). R _t 1.25 min (method 1a); m/z 262 [M+H]+ (ER+), 1H NMR (400 MHz, DMSO-0 ₆ ): δ, ppm 12.38 (brs, 1H), 9.51 (s, 1H), 7.91 (d, J=1.8 Hz, 1H), 7.47 (d, J=16.0 Hz, 1H), 7.33 (d, J=1.9 Hz, 1H), 6, 33 (d, J=16.0 Hz, 1H), 6.25 (s, 1H), 3.08-3.00 (m, 2H), 1.15 (s, 6H).

108 109

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Step 7. (E)-N-((7-Amino-2-methylbenzofuran-3-yl)methyl)-3 -(3,3-dimethyl-4-oxo-2,3,4,5tetrahydro-III-pyrido [2,3-b][1,4]diazepin-8-yl)-N-methylacrylamide (compound 109).

DIPEA (0.5 mL, 2.9 mmol) was added dropwise to a stirred solution of 2-methyl-3((methylamino)methyl)benzofuran-7-amine 74 (0.04 g, 0.2 mmol) and trifluoroacetate (E )-3-(3,3-dimethyl-4oxo-2,3,4,5-tetrahydro-1H-pyrido[2,3-b][1,4]diazepin-8-yl)acrylic acid 108 (0, 07 g, 0.2 mmol) in DMF (2 ml) at RT. The reaction mixture was stirred for 10 minutes, then GATU (0.09 g, 0.2 mmol) was added and the reaction mixture was stirred for 16 hours. The reaction mixture was diluted with water (10 ml) and the precipitate was collected by filtration. The crude product was purified by column chromatography (0-5% MeOH/DCM) to give the title compound 109 as a yellow solid (19 mg, 24% yield). Rt 1.57 min (method 1a); m/z 434 [M+H]+ (ER ⁺ ), 1H NMR (400 MHz, DMSOC, 363 K): δ, ppm 8.77 (s, 1H), 7.92 (d, J=1 .9 Hz, 1H), 7.44 (d, J=15.4 Hz, 1H), 7.35 (d, J=2.0 Hz, 1H), 7.07 (d, J=15.6 Hz, 1H), 6.87 (t, J=7.7 Hz, 1H), 6.74 (d, J=7.3 Hz, 1H), 6.53 (d, J=7.6 Hz, 1H), 6.03-5.91 (m, 1H), 4.86 (s, 2H), 4.71 (s, 2H), 3.07 (d, J=4.6 Hz, 2H), 2.97 (s, 3H), 2.47 (s, 3H), 1.18 (s, 6H).

Example 18. Synthesis of (E)-3-(3-hydroxy-4'-oxo-1',2',4',5'-tetrahydrospiro[cyclobutane-1,3'-pyrido[2,3b][1, 4]diazepin]-8'-yl)-N-methyl-N-((3-methylbenzofuran-2-yl)methyl)acrylamide (compound 115).

General synthesis scheme.

Reaction conditions: a) 5-bromo-3-fluoro-2-nitropyridine 11, TEA, EtOH, reflux; b) Fe, NH4Cl, EtOH, H ₂ O, reflux; c) LiOH, H ₂ O, THF; d) GATU, DIPEA, DMF; e) Pd-162, MeNCy ₂ , NBu ₄ Cl, 1,4-dioxane, 80°C.

Step 1. Methyl 1-(((5-bromo-2-nitropyridin-3-yl)amino)methyl)-3-hydroxycyclobutanecarboxylate (compound 111).

To a solution of 5-bromo-3-fluoro-2-nitropyridine 11 (230 mg, 1.0 mmol) in ethanol (10 ml) add methyl 1-(aminomethyl)-3-hydroxycyclobutanecarboxylate hydrochloride 110 (Enamine) (200 mg, 1 .0 mmol) and TEA (0.6 ml, 4.1 mmol). The reaction mixture is heated to reflux overnight. The solvent is removed in vacuo and the reaction mixture is partitioned between water (2 ml) and ethyl acetate (2 ml). The organic extract is separated, dried with MgSO ₄ , filtered and concentrated in vacuo. The crude product was purified by chromatography (0-100% EtOAc/isohexane) to give the title compound 111 as a yellow oil (250 mg, 61% yield). Rt 1.58 min (method 1a); m/z 360/362 [M+H] ⁺ (ER ⁺ ), 1H NMR (400 MHz, CDCh): δ, ppm 8.10 (s, 1H), 7.91 (d, J=1, 9 Hz, 1H), 7.63 (d, J=1.9 Hz, 1H), 4.58 (tt, J=7.2 Hz, 5.8 Hz, 1H), 3.81 (s, 3H ), 3.72 (d, J=5.6 Hz, 2H), 2.93-2.88 (m, 2H), 2.10-2.06 (m, 2H) (omitted OH).

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Step 2. Methyl 1-(((2-amino-5-bromopyridin-3-yl)amino)methyl)-3hydroxycyclobutane carboxylate (compound 112).

A mixture of methyl 1 -(((5-bromo-2-nitropyridin-3 -yl)amino)methyl)-3-hydroxycyclobutanecarboxylate 111 (245 mg, 0.7 mmol), iron powder (152 mg, 2.7 mmol) and Ammonium chloride (364 mg, 6.8 mmol) in a solvent mixture of ethanol (12 ml) and water (3 ml) is heated at reflux for 1 hour. The mixture is filtered hot through a pad of Celite®, then concentrated in vacuo. The residue is partitioned between water (20 ml) and DCM (20 ml), and the aqueous layer is extracted with DCM (2x20 ml). The combined organic layers are dried with MgSO ₄ , filtered and concentrated in vacuo. The crude product was purified by chromatography (0-10% MeOH/DCM) to give the title compound 112 as a yellow oil (180 mg, 76%). Rt 0.91 min (method 1a); m/z 330/332 [M+H]+ (ER ⁺ ).

he he

112 113

Step 3. 1-(((2-Amino-5-bromopyridin-3-yl)amino)methyl)-3-hydroxycyclobutanecarboxylic acid (compound 113).

A mixture of methyl 1-(((2-amino-5-bromopyridin-3-yl)amino)methyl)-3-hydroxycyclobutanecarboxylate 112 (175 mg, 0.5 mmol) and lithium hydroxide (66 mg, 2.7 mmol) in solvent mixtures of THF (4.5 ml) and water (1.5 ml) were stirred at RT for 3 hours. The reaction mixture was then acidified to pH 3 by adding acetic acid (~0.4 ml). The solvent was removed in vacuo to give the title compound 113 as a yellow solid (190 mg, 98% yield), which was used in the next step without further purification. Rt 0.53 min (method 1a); m/z 316/318 [M+H] ⁺ (ER ⁺ ).

He

113 114

Step 4. 8'-Bromo-3-hydroxy-1',2'-dihydrospiro[cyclobutane-1,3'-pyrido[2,3-b][1,4]diazepine]4'(5'H)- he (connection 114).

K-ethyl- Kisopropylpropan-2-amine (0.5 ml, 2.7 mmol), then GATU (250 mg, 0.7 mmol). The reaction mixture was stirred at RT for 1 hour. Water (4 ml) was then added and the resulting mixture was extracted with DCM (3x5 ml). The combined organic phases are washed with brine (5 ml), dried with MgSO ₄ , filtered and concentrated in vacuo. The crude product was purified by chromatography (010% MeOH/DCM) to give the title compound 114 as a colorless solid (30 mg, 17% yield). Rt 1.20 min (method 1a); m/z 298/300 [M+H] ⁺ (ER ⁺ ), 1H NMR (400 MHz, CD ₃ OD): δ, ppm 7.72 (d, J=2.0 Hz, 1H), 7 .27 (d, J=2.0 Hz, 1H), 4.28 (p, J=7.2 Hz, 1H), 3.46 (s, 2H), 2.85-2.66 (m, 2H), 1.94-1.79 (m, 2H).

114 115

Step 5. (E)-3-(3-Hydroxy-4'-oxo-1',2',4',5'-tetrahydrospiro[cyclobutane-1,3'-pyrido[2,3b][1,4 ]diazepin]-8'-yl)-K-methyl-K-((3-methylbenzofuran-2-yl)methyl)acrylamide (compound 115).

K-metl-K-((3-methylbenzofuran-2-yl)methyl)acrylamide 9 (25 mg, 0.1 mmol), 8'-bromo-3-hydroxy-1',2'-dihydrospiro is loaded into the reaction tube [cyclobutan-1,3'-pyrido[2,3-b][1,4]diazepine]4'(5Ή)-one 114 (28 mg, 0.1 mmol), Bu ₄ NCl (3 mg, 9. 4 µmol), [P(tBu)3]Pd(crotyl)Cl (Pd-162) (4 mg, 9.4 µmol). The tube is then flushed with nitrogen for 5 minutes. Add 1,4-dioxane (2.5 ml) and N

- 77 046887 cyclohexyl-N-methylcyclohexanamine (41 µl, 0.2 mmol) and the reaction mixture was purged with nitrogen for a further 5 minutes. The mixture is heated to 80°C for 1 hour and cooled to room temperature. Ethyl acetate (5 ml) was added and the resulting solution was washed with H ₂ O (5 ml), brine (5 ml), dried over MgSO ₄ , filtered and concentrated in vacuo. The crude product was purified by column chromatography (0-10% MeOH/DCM) to give the title compound 115 as a pale yellow solid (22 mg, 49%). Rt 1.79 min (method 1a); m/z 447 [M+H]+ (ER ⁺ ), 1H NMR (400 MHz, DMSOC): δ, ppm 9.05 (s, 1H), 7.94 (d, J=1.9 Hz , 1H), 7.56 (dd, J=7.6, 1.4 Hz, 1H), 7.47 (d, J=8.0 Hz, 1H), 7.45-7.34 (m, 2H), 7.33-7.19 (m, 2H), 7.10 (d, J=15.6 Hz, 1H), 5.97 (d, J=4.7 Hz, 1H), 4, 84 (s, 3H), 4.21-4.03 (m, 1H), 3.41 (d, J=3.9 Hz, 2H), 3.10 (s, 3H), 2.64-2 .54 (m, 2H), 2.28 (s, 3H), 1.83-1.74 (m, 2H).

Example 19. Synthesis of (E)-N-((7-(benzo^][1,3]dioxol-5-yloxy)-2-methylbenzofuran-3-yl)methyl)-Nmethyl-3-(4-oxo- 2,3,4,5-tetrahydro-1H-pyrido[2,3-b][1,4]diazepin-8-yl)acrylamide (compound 119).

General synthesis scheme.

/

118 119 o T

Reaction conditions: a) CuI, 2-(dimethylamino)acetic acid, Cs ₂ CO ₃ , DMF, 110°C; b) 1chloroethyl chloroformate, DCM, 0°C to RT; c) GATU, DIPEA, DMF. /

Step 1. 1-(7-(Benzo^][1,3]dioxol-5-yloxy)-2-methylbenzofuran-3-yl)-N-(4-methoxybenzyl)-Nmethylmethanamine (compound 117).

Mixture of 3-(((4-methoxybenzyl)(methyl)amino)methyl)-2-methylbenzofuran-7-ol 76 (150 mg, 0.48 mmol), 1-iodo3,4-methylenedioxybenzene 116 (Fluorochem) (358 mg , 1.45 mmol), 2-(dimethylamino)acetic acid (50 mg, 0.48 mmol), copper(1) iodide (92 mg, 0.48 mmol) and cesium carbonate (942 mg, 2.89 mmol) evacuate and fill back with nitrogen (3 times).

DMF (3 ml) is added and the mixture is heated to 110°C overnight. The reaction mixture is cooled to RT, poured into water (30 ml) and the crude product is extracted into ethyl acetate (2x30 ml). The organic extracts are combined and dried over Na ₂ SO ₄ . The crude product was purified by chromatography [0-10% (0.7 M ammonia/MeOH)/DCM] to give the title compound 117 as a clear yellow gum (85 mg, 38% yield). Rt 1.69 min (method 1a) m/z 432 [M+H] ⁺ (ER ⁺ ).

/

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Step 2. 1-(7-(BenzoI[1,3]dioxol-5-yloxy)-2-methylbenzofuran-3-yl)-Nmethylmethanamine (compound 118).

To a solution of 1-(7-(benzo^][1,3]dioxol-5-yloxy)-2-methylbenzofuran-3-yl)-N(4-methoxybenzyl)-N methylmethanamine 117 (85 mg, 0.20 mmol ) in DCM (2 ml) add 1-chloroethylcarbonochloridate (0.031 ml, 0.29 mmol) at 0°C and the mixture is allowed to reach RT, then stir for 1 hour. The mixture is evaporated to dryness and the residue is taken up in methanol (5 ml ) and the mixture is heated to reflux for 1 hour. The reaction mixture is cooled to RT and the solution is applied directly to an SCC column (1 g). The column was washed with methanol (10 mL) and the crude product was eluted with 10% methanol ammonia (10 mL) to give the title compound 118 as a yellow oil (60 mg, 90% yield). Rt 1.38 min (method 1a) m/z 281[M+H]+ (ER+).

Step 3. (E)-N-((7-(Benzo[d][1,3]dioxol-5-yloxy)-2-methylbenzofuran-3-yl)methyl)-N-methyl-3-(4oxo- 2,3,4,5-tetrahydro-1H-pyrido[2,3-b][1,4]diazepin-8-yl)acrylamide (compound 119).

Solution of 1-(7-(benzo^][1,3]dioxol-5-yloxy)-2-methylbenzofuran-3-yl)-Nmethylmethanamine 118 (60 mg, 0.19 mmol), trifluoroacetate (E)-3 -(4-oxo-2,3,4,5-tetrahydro-1H-pyrido[2,3-b][1,4]diazepin-8yl)acrylic acid 9 (67 mg, 0.19 mmol) and DIPEA ( 135 μl, 0.77 mmol) in DMF (3 ml) add GATU (73 mg, 0.19 mmol) and the mixture is stirred at RT for 1 hour. The reaction mixture is quenched by adding water (10 ml). The solid is collected by filtration and dried.

The crude product is initially purified by chromatography (0-10% MeOH/DCM) to give the title compound 119 as a pale yellow solid (LCMS purity 93%). The solid was then purified by trituration with acetonitrile (10 ml), then further triturated with DMF (2 ml) and washed with TBME (10 ml) to obtain the title compound 119 as a pale yellow solid (14 mg, 13% yield). R _t 2.12 min (method 1a) m/z 527[M+H]+ (ER+), ¹ H NMR (500 MHz, DMSO-de, 363 K): δ, ppm 9.14 (s , 1H), 7.98 (d, J=1.9 Hz, 1H), 7.46 (d, J=15.4 Hz, 1H), 7.38 (d, J=1.9 Hz, 1H ), 7.30 (d, J=7.8 Hz, 1H), 7.15-7.06 (m, 2H), 6.85 (d, J=8.4 Hz, 1H), 6.78 (dd, J=8.0, 1.0 Hz, 1H), 6.69 (d, J=2.5 Hz, 1H), 6.50 (dd, J=8.4, 2.5 Hz, 1H), 6.01 (s, 2H), 5.80 (s, 1H), 4.77 (s, 2H), 3.44 (dd, J=6.9, 3.5 Hz, 2H), 2.67-2.62 (m, 2H), 1.20 (s, 1H). Both CH3 protons are darkened by solvent and water.

Example 20. Synthesis of 3-(4-((3rS,6rR)-2-((E)-3-(4-oxo-2,3,4,5-tetrahydro-1H-nirido[2,3-b] [1,4]diazenin-8yl)acryloyl)-1,2,3,3g,4,6g-hexahydrocyclopenta[e]pyrrol-5-yl)phenoxy)benzonitrile (compound 123). General synthesis scheme.

Reaction conditions: a) K ₂ CO ₃ , H ₂ O, PdCl ₂ (dppf), 1,4-dioxane, 80°C; b) TPA, DXM; c) GATU, DIPEA, DMF.

The synthesis of 3-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenoxy)benzonitrile (compound 120) was previously described in Abbott Laboratories - US2002/156081, 2002, A1. This compound is also commercially available from Fluorochem (CAS: 330792-98-8).

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Stage 1. tert-Butyl

(3r6,6G)-5-(4-(3-cyanophenoxy)phenyl)-3,3g,4,6g-tetrahydrocyclopenta^pyrrole^Nfcarboxylate (compound 121).

Mixture of 3-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenoxy)benzonitrile 120 (250 mg, 0.78 mmol), tert-butyl 5-(( (trifluoromethyl)sulfonyl)oxy)-3,3r,6,6r-tetrahydrocyclopenta[c]pyrrole2(III)-carboxylate) 57 (278 mg, 0.8 mmol), PdCl ₂ (dppf)-CH ₂ Cl ₂ adduct ( 64 mg, 0.08 mmol), potassium carbonate (323 mg, 2.3 mmol) and water (1.5 ml) in 1,4-dioxane (4 ml) were stirred at 80°C overnight. The reaction mixture is cooled to RT, filtered through Celite® and washed with EtOAc (50 ml). The mixture was then evaporated in vacuo to give the title compound 121 as a brown oil (396 mg, qty) which was used at the next Rt 2.86 min (method 1b); m/z 347 [M+H-tBu]+ (ER ⁺ ).

stage without further purification.

Step 2. 3-(4-((3rS,6rR)-1,2,3,3r,4,6r-Hexahydrocyclopenta[c]pyrrol-5-yl)phenoxy)benzonitrile (compound 122).

Trifluoroacetic acid (2.5 ml) is added dropwise to the stirred solution of tert-butyl 5-(4-(3-cyanophenoxy)phenyl)-3,3r,6,6r-tetrahydrocyclopenta[c]pyrrole-2(1H)-carboxylate 121 (313 mg, 0.8 mmol) in DXM (2.5 mL) at RT. The reaction mixture was stirred for 2 hours, then the solvent was concentrated in vacuo and the resulting residue was taken up in methanol (30 ml) and applied to an SCC column. The column is washed with methanol (50 ml) and the product is eluted with a solution of 10% methanolic ammonia. The mixture was then evaporated to give the title compound 122 as a yellow oil (154 mg, 64% yield). Rt 1.75 min (method 1b); m/z 303 [M+H] ⁺ (ER ⁺ ), ¹ H NMR (400 MHz, CDCl ₃ ): δ, ppm 7.44-7.39 (m, 3H), 7.34 (d, J=7.6 Hz, 1H), 7.24-7.18 (m, 2H), 6.97 (d, J=8.5 Hz, 2H), 5.93 (d, J=2.6 Hz, 1H), 3.50-3.45 (m, 1H), 3.06-2.98 (m, 2H), 2.97-2.90 (m, 2H), 2.90-2, 84 (m, 1H), 2.82-2.75 (m, 1H), 2.49 (m, 1H), 2.41 (shs, 1H).

Step 3. 3-(4-((3rS,6rR)-2-((E)-3-(4-Oxo-2,3,4,5-tetrahydro-1H-pyrido[2,3-b][ 1,4]diazepin-8yl)acryloyl)-1,2,3,3r,4,6r-hexahydrocyclopenta[c]pyrrol-5-yl)phenoxy)benzonitrile (compound 123).

Suspension of 3-(4-((3rS,6rR)-1,2,3,3r,4,6r-hexahydrocyclopenta[c]pyrrol-5-yl)phenoxy)benzonitrile 122 (70 mg, 0.2 mmol), trifluoroacetate (E)-3-(4-oxo-2,3,4,5-tetrahydro-1H-pyrido[2,3-b][1,4]diazepin8-yl)acrylic acid 6 (76 mg, 0.2 mmol), Hetyl-U-isopropylpropan-2-amine (0.20 ml, 1.2 mmol) in DMF (1 ml) was stirred for 10 min. GATU (88 mg, 0.2 mmol) was added in one portion and the reaction mixture was stirred at room temperature for 1 hour. The mixture was diluted with water (10 ml) and the precipitate was collected by filtration. The crude product was purified by column chromatography (0-5% MeOH/DCM) to give the title compound 123 as a yellow solid (87 mg, 72% yield). Rt 2.08 min (method 1b); m/z 518 [M+H] ⁺ (ER ⁺ ), ¹ H NMR (400 MHz, DMSO-dg): δ, ppm 9.74 (d, J=6.9 Hz, 1H), 7, 98 (dd, J=8.4 Hz, 1.9 Hz, 1H), 7.60-7.53 (m, 4H), 7.50-7.47 (m, 1H), 7.37-7 .31 (m, 3H), 7.09-7.04 (m, 2H), 6.87 (dd, J=15.5 Hz, 9.0 Hz, 1H), 6.18 (d, J= 4.7 Hz, 1H), 6.05-6.00 (m, 1H), 4.00 (t, J=9.9 Hz, 1H), 3.87-3.79 (m, 2H), 3.69-3.51 (m, 2H), 3.43-3.37 (m, 2H), 3.19-3.12 (m, 1H), 3.03-2.90 (m, 2H ), 2.66-2.59 (m, 3H).

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Example 21. Synthesis of 8-((E)-3-((3r6,6G)-5-(4-(3-methoxyphenoxy)phenyl)-3.3g,4.6g-tetrahydrocyclopenta[c]pyrrole-2( 1 H)-yl)-3-oxoprop-1-en-1-yl)-1,2,3,5-tetrahydro-4H-pyrido[2,3-b][1,4]diazepin-4-one ( connection 129).

General synthesis scheme.

Reaction conditions: a) 1-bromo-4-iodobenzene 125, CuI, U,U-dimethylglycine, Cs ₂ CO ₃ , DMF, 110°C; b) K ₂ CO ₃ , H ₂ O, PdCl ₂ (dppf), 1,4-dioxane, 80°C; c) TPA, DXM; d) GATU, DIPEA, DMF.

Step 1. 1-(4-Bromophenoxy)-3-methoxybenzene and 1-(4-iodophenoxy)-3-methoxybenzene (compound 126).

Mixture of 1-bromo-4-iodobenzene 125 (Sigma Aldrich) (500 mg, 1.8 mmol), copper iodide (337 mg, 1.8 mmol), U,U-dimethylglycine (182 mg, 1.8 mmol) , cesium carbonate (2.30 g, 7.1 mmol) and 3 methoxyphenol 124 (Sigma Aldrich) (0.19 ml, 1.8 mmol) were evacuated and backfilled with nitrogen (3 times). DMF (10 ml) is added and the mixture is heated to 110°C overnight. The mixture is filtered through a pad of Celite®, washing with EtOAc (50 ml) and then evaporated to dryness. The crude product was purified by column chromatography (0-10% EtOAc/isohexane) to give the mixture of title compounds 126 as a dark oil (338 mg, 53% yield), which was used in the next step without further purification, 1H NMR (400 MHz, CDCl ₃ ): δ, ppm 7.66-7.62 (m, 1H), 7.51-7.43 (m, 3H), 7.28-7.24 (m, 2H), 6, 96-6.89 (m, 3H), 6.82-6.79 (m, 1H), 6.71 (ddt, J=8.3 Hz, 2.2 Hz, 1.3 Hz, 2H), 6.60 (dkv, J=9.4 Hz, 1.3 Hz, 4H), 3.81 (s, 6H).

126 127

Step 2. tert-Butyl (3rS,6rR)-5-(4-(3-methoxyphenoxy)phenyl)-3,3r,4,6r-tetrahydrocyclopenta[c]pyrrole-2(1H)-carboxylate (compound 127) .

A mixture of 1-(4-iodophenoxy)-3-methoxybenzene 126 (169 mg, 0.51 mmol), 1-(4-bromophenoxy)-3methoxybenzene 126 (169 mg, 0.6 mmol), tert-butyl 5-(4 ,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)3,3r,6,6r-tetrahydrocyclopenta[c]pyrrole-2(1H)-carboxylate 86 (375 mg, 1.1 mmol ), the adduct of PdCl ₂ (dppf)CH ₂ Cl ₂ (91 mg, 0.1 mmol), potassium carbonate (464 mg, 3.4 mmol) and water (2 ml) in 1,4-dioxane (6 ml) is stirred at 80°C for 4 hours. The reaction mixture is cooled to RT, filtered through Celite® and washed with EtOAc (50 ml). The mixture is then evaporated under vacuum. The crude product was purified by column chromatography (0-50% EtOAc/isohexane) to give the title compound 127 as a white solid (209 mg, 45% yield). R _t 3.00 min (method 1b); m/z 352 [M+H-tBu]+

- 81 046887 (ER ⁺ ), 1H NMR (400 MHz, CDC13): δ, ppm 7.44-7.38 (m, 2H), 7.23 (td, J=8.2 Hz, 7, 4 Hz, 1.2 Hz, 1H), 7.036.96 (m, 2H), 6.69-6.66 (m, 1H), 6.63-6.57 (m, 2H), 5.97 ( shs, 1H), 3.79 (d, J=2.1 Hz, 3H), 3.75-3.69 (m, 1H), 3.59-3.48 (m, 3H), 3.11 (dd, J=11.0 Hz, 6.9 Hz, 1H), 3.06-2.99 (m, 1H), 2.97-2.90 (m, 1H), 2.59 (d, J=15.7 Hz, 1H), 1.47 (s, 9H).

127 128

Step 3. (3rS,6rR)-5-(4-(3-Methoxyphenoxy)phenyl)-1,2,3,3r,4,6r-hexahydrocyclopenta[c]pyrrole (compound 128).

Trifluoroacetic acid (1 ml) was added dropwise to a stirred solution of tert-butyl 5(4-(3-methoxyphenoxy)phenyl)-3,3r,6,6r-tetrahydrocyclopenta[c]pyrrole-2(1H)-carboxylate 127 (209 mg, 0.5 mmol) in DCM (1 ml) at RT. The reaction mixture is stirred for 1 hour, then the solvent is concentrated in vacuo. The resulting residue is placed in methanol (30 ml) and applied to an SCC column. The column is washed with methanol (50 ml) and the product is eluted with a solution of 10% methanolic ammonia. The mixture was then evaporated to give the title compound 128 as a yellow oil (157 mg, 96% yield). Rt 1.91 min (method 1b); m/z 308 [M+H] ⁺ (ER ⁺ ), 1H NMR (CDCl ₃ ): δ, ppm 7.37 (d, J=8.5 Hz, 2H), 7.23-7.17 (m, 1H), 6.99-6.93 (m, 2H), 6.67-6.62 (m, 1H), 6.59-6.55 (m, 2H), 5.89-5 .87 (m, 1H), 3.75 (s, 3H), 3.47-3.42 (m, 3H), 3.05-2.78 (m, 5H), 2.50-2.46 (m, 1H), 1.36 (d, J=8.3 Hz, 1H).

128 129

Step 4. (3rS,6rR)-5-(4-(3-Methoxyphenoxy)phenyl)-1,2,3,3r,4,6r-hexahydrocyclopenta[c]pyrrole (compound 129).

Suspension of (3rS,6rR)-5-(4-(3-methoxyphenoxy)phenyl)-1,2,3,3r,4,6r-hexahydrocyclopenta[c]pyrrole 128 (70 mg, 0.3 mmol), trifluoroacetate ( E)-3-(4-oxo-2,3,4,5-tetrahydro-1H-pyrido[2,3-b][1,4]diazepin8-yl)acrylic acid 6 (75 mg, 0.3 mmol ), X-ethyl-X-isopropylpropan-2-amine (0.2 ml, 1.2 mmol) in DMF (1.5 ml) was stirred for 10 min. GATU (87 mg, 0.3 mmol) was added in one portion and the reaction mixture was stirred at room temperature for 1 hour. The mixture was diluted with water (10 ml) and the precipitate was collected by filtration. The crude product was purified by column chromatography (0-5% MeOH/DCM) to give the title compound 129 as a yellow solid (86 mg, 70% yield). Rt 2.19 min (method 1b); m/z 523 [M+H] ⁺ (ER ⁺ ), 1H NMR (400 MHz, DMSO-de): δ, ppm 9.74 (d, J=6.8 Hz, 1H), 7.98 (dd, J=8.4 Hz, 1.9 Hz, 1H), 7.50 (d, J=8.5 Hz, 2H), 7.38-7.26 (m, 3H), 7.016.97 (m, 2H), 6.89-6.84 (m, 1H), 6.72 (dd, J=8.3 Hz, 2.4 Hz, 1H), 6.59-6.58 (m, 1H), 6.56-6.53 (m, 1H), 6.14-6.13 (m, 1H), 6.05-5.98 (m, 1H), 3.99 (t, J= 9.8 Hz, 1H), 3.87-3.78 (m, 1H), 3.73 (d, J=0.8 Hz, 3H), 3.68-3.51 (m, 2H), 3.42 (td, J=6.1 Hz, 3.4 Hz, 2H), 3.20-3.11 (m, 1H), 3.02-2.87 (m, 2H), 2.61 (dt, J=10.9 Hz, 6.0 Hz, 3H).

- 82 046887

Example 22. Synthesis of (E)-N-methyl-N-((2-methyl-7-((1-methylindolin-6-yl)oxy)benzofuran-3yl)methyl)-3-(4-oxo-2, 3,4,5-tetrahydro-1H-pyrido[2,3-b][1,4]diazepin-8-yl)acrylamide (compound 134).

General synthesis scheme.

Reaction conditions: a) NaH, MeI, THF, 0°C; b) CuI, K,K-dimethylglycine, Cs ₂ CO ₃ , DMF, 110°C; c) TPA, DXM; d) GATU, DIPEA, DMF.

Step 1. 6-Bromo-1-methylindoline (compound 131).

Sodium hydride (60% in mineral oil, 0.15 g, 3.8 mmol) is added in small portions to a stirred solution of 6-bromoindoline 130 (0.5 g, 2.5 mmol) in THF (20 ml) at 0°C under N2. The reaction mixture was stirred for 30 min, then iodomethane (0.2 ml, 2.5 mmol) was added and the reaction mixture was warmed to RT. The reaction mixture is stirred at RT over the weekend, then cooled to 0°C and quenched by adding sat. NH4Cl (50 ml). The aqueous mixture was extracted with EtOAc (3x50 ml), and the combined organic extracts were washed with brine (50 ml), dried with MgSO ₄ , and concentrated in vacuo. The crude product was purified by column chromatography (0-20% EtOAc/isohexane) to give the title compound 131 as a brown oil (0.44 g, 76%). R _t 2.39 min (method 1a) m/z 212/214 [M+H]+ (ER ⁺ ), ¹ H NMR (400 MHz, DMSO-dQ: δ, ppm 6.97-6.91 (m, 1H), 6.69 (dd, J=7.6 Hz, 1.8 Hz, 1H), 6.62 (d, J=1.7 Hz, 1H), 3.28 (t, J =8.3 Hz, 2H), 2.82 (t, J=8.5 Hz, 2H), 2.69 (s, 3H).

Step 2. tert-Butyl methyl ((2-methyl-7-((1-methylindolin-6-yl)oxy)benzofuran-3yl)methyl)carbamate (compound 132).

DMF (5 ml) is added to a mixture of Cs ₂ CO ₃ (0.67 g, 2.1 mmol), 6-bromo-1-methylindoline 131 (Fluorochem) (0.4 g, 1.9 mmol), tert-butyl ((7-hydroxy-2-methylbenzofuran-3-yl)methyl)(methyl)carbamate 98 (0.15 g, 0.5 mmol), 2-(dimethylamino)acetic acid (0.05 g, 0.5 mmol ) and CuI (0.1 g, 0.5 mmol) under N ₂ atmosphere, and the reaction mixture was heated to 110°C. The reaction mixture is stirred at this temperature for 16 hours, then the reaction mixture is cooled to RT. The crude mixture was loaded dry into Celite® and purified by column chromatography (0-50% EtOAc/isohexane) to give the title compound 132 as a colorless oil (41 mg, 19% yield). R _t 3.02 min (method 1a) m/z 423 [M+H] ⁺ (ER ⁺ ).

- 83 046887

Step 3. \-Methyl-1-(2-methyl-7-((1-methylindolin-6-yl)oxy)benzofu ^; ran-3yl)methanamine trifluoroacetate (compound 133).

Trifluoroacetic acid (2 ml) is added dropwise to a stirred solution of tert-butyl methyl ((2-methyl-7-((1-methylindolin-6-yl)oxy)benzofuran-3-yl)methyl)carbamate 132 (0.04 g, 0.1 mmol) in DCM (2 ml) at RT. The reaction mixture was stirred for 30 minutes, then concentrated in vacuo to give the title compound 133 as a brown oil which was used in the next step without further purification. Rt 2.02 min (method 1a) m/z 323 [M+H] (ER).

Step 4. (E)-N-Methyl-N-((2-methyl-7-((1-methylindolin-6-yl)oxy)benzofuran-3-yl)methyl)-3-(4oxo-2,3 ,4,5-tetrahydro-1H-pyrido[2,3-b][1,4]diazepin-8-yl)acrylamide (compound 134).

DIPEA (0.5 mL, 2.9 mmol) is added dropwise to a stirred solution of Nmethyl-1-(2-methyl-7-((1-methylindolin-6-yl)oxy)benzofuran-3-yl)methanamine trifluoroacetate 133 (0.07 g, 0.2 mmol) and trifluoroacetate (E)-3-(4-oxo-2,3,4,5-tetrahydro-1H-pyrido[2,3-b][1,4]diazepine -8-yl)acrylic acid 6 (0.06 g, 0.2 mmol) in DMF (2 ml) at RT. The reaction mixture was stirred for 10 minutes, then GATU (0.08 g, 0.2 mmol) was added and the reaction mixture was stirred for 2 hours. The reaction mixture was diluted with H2O (10 ml) and the precipitate was collected by filtration.

The crude product was purified by column chromatography (0-3% MeOH/DCM) to give the title compound 134 as a pale yellow solid (20 mg, 23% yield). Rt 2.13 min (method 1a) m/z 538 [M+H] ⁺ (ER ⁺ ), ¹ H NMR (400 MHz, 363 K, DMSO-dQ: δ, ppm 9.14 (s, 1H ), 7.98 (d, J=1.9 Hz, 1H), 7.46 (d, J=15.4 Hz, 1H), 7.38 (d, J=1.9 Hz, 1H), 7.28 (d, J=7.8 Hz, 1H), 7.16-7.04 (m, 2H), 6.95 (dt, J=7.8, 1.1 Hz, 1H), 6 .78 (dd, J=8.0, 1.1 Hz, 1H), 6.21-6.13 (m, 2H), 5.79 (s, 1H), 4.77 (s, 2H), 3.46-3.41 (m, 2H), 3.33 (t, J=8.2 Hz, 2H), 3.02 (s, 3H), 2.87 (t, J=8.2 Hz , 2H), 2.67 (s, 3H), 2.66-2.62 (m, 2H), 2.49 (s, 3H).

Example 23. Synthesis of 2-methoxy-4-(4-((3rS,6rR)-2-((E)-3-(4-oxo-2,3,4,5-tetrahydro-1H-pyrido[2, 3b][1,4]diazepin-8-yl)acryloyl)-1,2,3,3r,4,6r-hexahydrocyclopenta[c]pyrrol-5-yl)phenoxy)benzonitrile (compound 140).

General synthesis scheme.

Reaction conditions: a) 4-fluoro-2-methoxybenzonitrile 136, Cs ₂ CO ₃ , NMP, 80°C; b) Pd(dppf)Cl ₂ , K ₂ CO ₃ ,

H2O, 1,4-dioxane, 90°C; c) TPA, DXM; d) GATU, DIPEA, DMF.

CS2CO3,

NMP, 80 °C

135 136 137

- 84 046887

Step 1. 4-(4-Bromophenoxy)-2-methoxybenzonitrile (compound 137).

A mixture of 4-fluoro-2-methoxybenzonitrile 136 (Fluorochem) (1 g, 6.6 mmol), 4-bromophenol 135 (1.37 g, 7.94 mmol) and Cs ₂ CO ₃ (6.47 g, 19, 9 mmol) in X%1ethyl-2-11irrolidone (20 ml) is heated at 80°C for 2 hours. The reaction mixture is cooled to room temperature and poured into water (50 ml). The mixture is extracted with ethyl acetate (2x50 ml). The organics are combined and washed with brine (2x50 ml), passed through a hydrophobic frit, and concentrated in vacuo. The crude product was purified by column chromatography (0-50% EtOAc/isohexane) to give the title compound 137 as a clear, colorless oil that crystallized on standing (2.02 g, 70% yield). R _t 2.56 min (method 1a) m/z 304/306 [M+H]+ (ER+).

137 138

Step 2. tert-Butyl (3r6L)-5-(4-(4-cyano-3-methoxyphenoxy)phenyl)-3.3g,4.6gtetrahydrocyclopenta[c]pyrrole-2(1H)-carboxylate (compound 138 ).

Mixture of 4-(4-bromophenoxy)-2-methoxybenzonitrile 137 (0.18 g, 0.6 mmol), (3R'I,61X)-tert-butide 5(4,4,5,5-tetramethyl-1, 3,2-dioxaborolan-2-yl)-3.3g,6.6g-tetrahydrocyclopenta[c]pyrrole-2(1H)-carboxylate 86 (0.2 g, 0.6 mmol), K ₂ CO ₃ (0 .25 g, 1.8 mmol) in water (1 ml) and the PdCl ₂ (dppf)-CH ₂ Cl ₂ adduct (0.05 g, 0.06 mmol) in 1,4-dioxane (4 ml) degassed with nitrogen and then stirred at 80°C overnight. The reaction mixture is cooled to room temperature, diluted with water (10 ml), extracted with ethyl acetate (2x10 ml). The combined organic layers were washed with brine (20 ml), dried (MgSO ₄ ), filtered and concentrated in vacuo. The crude product was purified by column chromatography (0-50% EtOAc/isohexane) to give the title compound 138 as a thick, colorless oil (0.23 g, 73% yield). R _t 2.93 min (method 1a) m/z 377 [M+H-tBu]+ (ER+), 1H NMR (400 MHz, CDCl ₃ ): δ, ppm 7.51-7.44 ( m, 3H), 7.07-7.02 (m, 2H), 6.59 (d, J=2.2 Hz, 1H), 6.53 (dd, J=8.6 Hz, 2.2 Hz, 1H), 6.06-6.01 (m, 1H), 3.88 (s, 3H), 3.77-3.68 (m, 1H), 3.59-3.48 (m, 3H), 3.12 (dd, J=11.0 Hz, 6.9 Hz, 1H), 3.10-2.93 (m, 2H), 2.62 (d, J=15.7 Hz, 1H), 1.47 (s, 9H).

138 13E

Step 3. 4-(4-((3r6,6L)-1,2,3,3g,4,6g-Hexahydrocyclopenta[c]pyrrol-5-yl)phenoxy)-2-methoxybenzonitrile (compound 139).

To a stirred solution of (3gI,61X)-tert-butyl 5-(4-(4-cyano-3-methoxyphenoxy)phenyl)3.3g,6.6g-tetrahydrocyclopenta[c]pyrrole-2(III)-carboxylate 138 ( 0.23 g, 0.5 mmol) in DCM (2 ml) add TFA (1.6 ml). The reaction mixture is stirred at room temperature for 1 hour. The solvent is removed in vacuo. The resulting oil was taken up in MeOH (5 ml) and applied to an SCC column. The column was washed with methanol (5 mL) and the product was eluted with 10% methanolic ammonia (5 mL) to give the title compound 139 as a colorless crystalline solid (0.1 g, 56%). R _t 1.50 min (method 1a) m/z 333 [M+H]+ (ER ⁺ ).

139 140

Step 4. 2-Methoxy-4-(4-((3r6L)-2-((E)-3-(4-oxo-2,3,4,5-tetrahydro-III-pyrido[2,3b ][1,4]diazepin-8-yl)acryloyl)-1,2,3,3g,4,6g-hexahydrocyclopenta[c]pyrrol-5-yl)phenoxy)benzonitrile (compound 140).

To a stirred solution of 4-(4-((3r6,6L)-1,2,3,3g,4,6g-hexahydrocyclopenta[c]pyrrol-5yl)phenoxy)-2-methoxybenzonitrile (0.1 g, 0.29 mmol) in DMF (2 ml) add trifluoroacetate (E)-3-(4oxo-2,3,4,5-tetrahydro-III-pyrido[2,3-b][1,4]diazepin-8-yl) acrylic acid (75 mg, 0.32 mmol) and Nethyl-X-isopropylpropan-2-amine (0.26 ml, 1.46 mmol). The reaction mixture was stirred at RT for 5 min and GATU (0.1 g, 0.35 mmol) was added. The reaction mixture is stirred for 1 hour. Water (2 ml) is added and the aqueous phase is extracted with DCM (3x5 ml). Combined organic phases

- 85 046887 washed with H2O (2x5 ml), brine (5 ml), dried (MgSO ₄ ), filtered and concentrated in vacuo. The crude solid was purified by column chromatography (0-10% MeOH/DCM) to give the title compound 140 as a yellow solid (0.1 g, 62% yield). Rt 2.11 min (method 1a) m/z 548 [M+H]+ (ER+), ¹ H NMR (400 MHz, DMSO-dg): δ, ppm 9.74 (d, J=6 .7 Hz, 1H), 7.98 (dd, J=8.3, 1.9 Hz, 1H), 7.69 (d, J=8.6 Hz, 1H), 7.60-7.55 (m, 2H), 7.39-7.29 (m, 2H), 7.15-7.11 (m, 2H), 6.91-6.83 (m, 2H), 6.53 (ddd , J=8.5 Hz, 6.1 Hz, 2.2 Hz, 1H), 6.20 (t, J=3.2 Hz, 1H), 6.02 (d, J=13.2 Hz, 1H), 4.05-3.96 (m, 0.5H), 3.90-3.76 (m, 4H), 3.70 (d, J=12.0 Hz, 0.5H), 3 .67-3.48 (m, 2H), 3.453.39 (m, 2H), 3.20-2.92 (m, 3H), 2.70-2.57 (m, 3H) (rotamers).

Example 24. Synthesis of (R, E)-N-((4-amino-3-methylbenzofuran-2-yl)methyl)-3-(3-hydroxy-3-methyl-4oxo-2,3,4,5- tetrahydro-III-pyrido[2,3-b][1,4]diazepin-8-yl)-X-methylacrylamide (compound 143).

General synthesis scheme.

Reaction conditions: a) chlorine(crotyl)(tri-tert-butylphosphine)palladium(P), [CH ₃ (CH ₂ ) ₃ ] ₄ NCl, DIPEA,

1,4-dioxane, 90°C, 18 h; b) TPA, DCM, 0°C to RT; c) HOBt-H ₂ O, EDC-HCl, DIPEA, DMF.

q ^^chloro(crotyl)(tri-tert-but1llphosphine)palladium(11) ⁰ [CH ₃ (CH ₂ )j]4NCI, DIPEA, 1,4-dioxane, 90 °C, 18 h

15 141

Stage 1. tert-Butyl (^E-3-(3-hydroxy-3-methyl-4-oxo-2,3,4,5-tetrahydro-III-pyrido[2,3b][1,4]diazepine- 8-yl)acrylate (compound 141).

A 100-ml flask is charged with (R)-8-bromo-3-hydroxy-3-methyl-1,2,3,5-tetrahydro-4H-pyrido[2,3ED^^diazepinX-one 47 (ChiralTek; chirality determined optional; 2.0 g, 7.2 mmol), n-Bu ₄ NCl (0.240 mg, 0.86 mmol), chloro(crotyl)(tri-tert-butylphosphine) palladium(P) (0.144 g, 0.36 mmol), and the mixture was purged with nitrogen for 10 minutes. tert-Butyl acrylate 15 (1.57 mL, 10.8 mmol), 1,4dioxane (52 mL) and DIPEA (2.45 mL, 14.4 mmol) were then added and nitrogen was bubbled through the mixture for a further 5 min. The reaction mixture is heated at 90°C for 18 hours and cooled to RT. The reaction mixture is filtered using 0.22 μM filters, washed with MeCN and concentrated to dryness in vacuo. Acetonitrile was added to the crude product, sonicated, and the yellow precipitate was filtered to give 141 (2.1 g, 6.5 mmol, 91.3% yield) as a yellow solid. R _t 1.45 min; LCMS: m/z: 321.3 [M+H]+. 1H NMR (400 MHz, DMSO-d6): δ, ppm 9.97 (s, 1H), 7.93 (d, J=1.7 Hz, 1H), 7.42 (d, J=16 ,1 Hz, 1H), 7.32 (d, J=1.7 Hz, 1H), 6.34 (d, J=16.1 Hz, 1H), 6.26 (s, 1H), 5, 31 (s, 1H), 3.15 (d, J=53.8 Hz, 2H), 1.47 (s, 9H), 1.23 (s, 3H).

141 142

Step 2. (R,E-3-(3-Hydroxy-3-methyl-4-oxo-2,3,4,5-tetrahydro-1H-pyrido[2,3-b][1,4]diazepine- 8yl)acrylic acid (compound 142).

To a solution of tert-butyl (^E-3-(3-hydroxy-3-methyl-4-oxo-2,3,4,5-tetrahydro-III-pyrido[2,3ED^^diazepin^-yl^acrylate 141 (0.364 g, 1.03 mmol) in DCM (9.1 ml) add 2,2,2 trifluoroacetic acid (0.117 g, 1.03 mmol) at 0°C. After completion of the reaction (UPLC-MS control), all solvents are removed under. DCM was then added and the resulting mixture was added to cold Et ₂ O. The yellow precipitate was filtered and washed with Et ₂ O to give the title compound 142 (0.287 g, 1.09 mmol, 106% yield) as a yellow solid. Rt 0.68 min; LCMS: m/z: 264.2 [M+H]+. 1H NMR (400 MHz, DMSO-d6): δ, ppm 12.39 (s, 1H), 9.98 (s, 1H), 7.92 (d, J=1.7 Hz, 1H), 7.47 (d, J=15.6 Hz, 1H), 7.32 (d, J=1.8 Hz , 1H), 6.35 (d, J=16.3 Hz, 1H), 3.23-3.09 (m, 3H), 1.23 (s, 3H).

- 86 046887

Step 3. (E)-N-[(4-Amino-3-methyl-benzofuran-2-yl)methyl]-3-[(3R)-3-hydroxy-3-methyl-4-oxo-2,5dihydro -1 H-pyrido[2,3 -b][1,4]diazepin-8-yl]-N-methylprop-2-enamide (compound 143).

In a microwave test tube covered with aluminum foil, a mixture of (E)-3-[(3R)-3-hydroxy-3methyl-4-oxo-2,5-dihydro-1H-pyrido[2,3-b][1,4 ]diazepin-8-yl]prop-2-enoic acid 142 (35 mg, 0.13 mmol) and 3-methyl-2-(methylaminomethyl)benzofuran-4-amine 3 (42.93 mg, 0.16 mmol) dissolve in DMF (0.22 ml). HOBt-H ₂ O (31.83 mg, 0.20 mmol) and EDC-HCl (39.84 mg, 0.20 mmol) were added to the reaction mixture and after 2 minutes of stirring at room temperature, DIPEA (0.08 ml , 0.46 mmol). The resulting mixture was stirred in the dark overnight at room temperature. The reaction mixture is diluted with acetonitrile, no precipitation is observed, but after a few more minutes a very fine solid appears. The mixture was diluted with DMF and purified by preparative HPLC and lyophilized to give 143 (25.7 mg, 0.0575 mmol, 43.72% yield) as a yellow solid. Rt 1.11 min; LCMS: m/z: 436.4 [M+H]+. ¹ H NMR (400 MHz, DMSOD ₆ ): δ, ppm 9.93 (s, 1H), 7.98 (d, J=11.6 Hz, 1H), 7.46-7.31 (m , 2.5H), 7.10-7.00 (m, 0.5H), 6.91 (t, J=7.9 Hz, 1H), 6.67-6.61 (m, 1H), 6.38 (d, J=7.9 Hz, 1H), 6.27-6.22 (m, 1H), 5.28 (s, 1H), 5.17 (shs, 2H), 4.83 (s, 0.8H, rotamer), 4.69 (s, 1.2H, rotamer), 3.23-3.07 (m, 4H), 2.92 (s, 1H), 2.46-2 .38 (m, 3H), 1.23 (s, 3H).

Example 25. Synthesis of (E)-3-((2R,3S)-3-hydroxy-2-methyl-4-oxo-2,3,4,5-tetrahydro-1H-pyrido[2,3b][1, 4]diazepin-8-yl)-N-methyl-N-((3-methylbenzofuran-2-yl)methyl)acrylamide (compound 150).

General synthesis scheme.

second enantiomer eluted

149

Reaction conditions: a) TEA, EtOH, reflux; b) Fe, NH ₄ Cl, EtOH, H2O, reflux; c) NaH, THF, 0°C to RT; d) chiral separation; e) Pd-162, MeNCy2, NBu ₄ Cl, 1,4-dioxane, 80°C.

144 11 145

Step 1. Ethyl 3-((5-bromo-2-nitropyridin-3-yl)amino)-2-hydroxybutanoate (compound 145).

TEA (2.8 mL, 20.1 mmol) was added to a stirred solution of 5-bromo-3-fluoro-2-nitropyridine 11 (1.1 g, 5.0 mmol) and ethyl 3-amino-2-hydroxybutanoate hydrochloride 144 (0.9 g, 4.9 mmol) in EtOH (20 ml) and the reaction mixture was heated to reflux for 4 hours. The reaction mixture was cooled to rt, then concentrated in vacuo, and the crude product was purified by column chromatography (DCM , then 0-10% MeOH/DCM) to give diastereomer 1 as yellow

- 87 046887 solid (0.55 g, 32%) and diastereomer 2 in the form of a yellow oil (0.50 g, 26%) together with another part which contains both diastereomers in the form of a yellow oil (0.22 g, 13%).

Diastereomer 1: Rt 1.76 min (method 1a) m/z 348/350 [M+H]+ (ER+). ^1H NMR (400 MHz, DMSO^ ₆ ): δ, ppm 8.04 (d, J=1.9 Hz, 1H), 7.97 (d, J=9.2 Hz, 1H), 7 .88 (d, J=1.8 Hz, 1H), 6.12 (d, J=4.8 Hz, 1H), 4.39-4.30 (m, 1H), 4.24 (dd, J=4.8 Hz, 2.8 Hz, 1H), 4.04 (kv, J=7.1 Hz, 2H), 1.24 (d, J=6.5 Hz, 3H), 1.09 (t, J=7.1 Hz, 3H).

Diastereomer 2: Rt 1.80 min (method 1a) m/z 348/350 [M+H]+ (ER+). ¹ H NMR (400 MHz, DMSO-06): δ, ppm 8.01 (d, J=1.9 Hz, 1H), 7.99 (d, J=8.9 Hz, 1H), 7 .90 (d, J=1.8 Hz, 1H), 6.09 (d, J=5.8 Hz, 1H), 4.28 (dd, J=5.9 Hz, 4.2 Hz, 1H ), 4.26-4.19 (m, 1H), 4.14 (kv, J=7.1 Hz, 2H), 1.21 (t, J=7.1 Hz, 3H), 1.14 (d, J=6.3 Hz, 3H).

145 146

Step 2. Ethyl 3-((2-amino-5-bromopyridin-3-yl)amino)-2-hydroxybutanoate (compound 146).

Mixture of ethyl 3-((5-bromo-2-nitropyridin-3-yl)amino)-2-hydroxybutanoate 145 (diastereomer 1) (0.55 g, 1.6 mmol), iron powder (0.71 g, 12 .6 mmol) and ammonium chloride (0.34 g, 6.3 mmol) in a mixture of solvents EtOH (50 ml) and H2O (10 ml) is stirred at reflux for 1 hour. The reaction mixture is loaded dry onto Celite® and purified by column chromatography (0-100% EtOAc/isohexane) to give the desired product 146 as a brown solid (0.28 g, 56%). Rt 0.99 min (method 1a) m/z 318/320 [M+H]+ (ER+). ¹ H NMR (400 MHz, DMSO-е ₆ ): δ, ppm, 7.28 (d, J=2.0 Hz, 1H), 6.72 (d, J=2.1 Hz, 1H) , 5.72 (s, 2H), 5.46 (d, J=6.7 Hz, 1H), 4.64 (d, J=9.2 Hz, 1H), 4.11 (dd, J= 6.7 Hz, 3.4 Hz, 1H), 4.08-3.95 (m, 2H), 3.88-3.76 (m, 1H), 1.21-1.05 (m, 6H ).

s he U _l

IBr-^χ^Ν ykxOB ^NaH _ Βγ^^,Ν-Ζ

Τ JI ο τγφ IJ >~ ^οη Ν ΝΗο Ν νΆ

Η Ο

146 147

Step 3. 8-Bromo-3-hydroxy-2-methyl-2,3-dihydro-1H-pyrido[2,3-b][1,4]diazepin-4(5H)-one (compound 147).

Sodium hydride (60% in mineral oil, 0.2 g, 5.00 mmol) is added in small portions to a stirred solution of ethyl 3-((2-amino-5-bromopyridin-3-yl)amino)-2-hydroxybutanoate 146 ( 1.0 g, 3.14 mmol) in THF (5.0 ml) at 0°C. The reaction mixture is warmed to RT and stirred for 4 hours. The reaction mixture is quenched by careful addition of NH ₄ Cl (50 ml) and the aqueous mixture is extracted with EtOAc (3x100 ml). The combined organic layers were washed with brine (1x100 mL), dried with MgSO ₄ , concentrated in vacuo and purified by column chromatography (0-50% EtOAc/isohexane) to give the desired product 147 as a brown solid (0.34 g, 37 %). Rt 1.18 min (method 1a) m/z 272/274 [M+H]+ (ER+). ¹ H NMR (500 MHz, DMSO-dQ: δ, ppm 10.23 (s, 1H), 7.70 (d, J=2.1 Hz, 1H), 7.29 (d, J=2 ,1 Hz, 1H), 6.36 (d, J=5.6 Hz, 1H), 5.18 (d, J=4.9 Hz, 1H), 4.18 (dd, J=4.8 Hz, 3.2 Hz, 1H), 3.79-3.65 (m, 1H), 1.08 (d, J=6.5 Hz, 3H).

Chiral resolution of compound 147.

n ✓

Βκ _Ζ χ .N-/ tg abs\ || I absV-QH nn—4

H o

H First enantiomer eluted\Chiral section 148

1Λ/ ^he '

Η;

Η Ο Κζχ,/Νχ·

147 ^Ν Η Ό

Second enantiomer eluted

149

Step 4. (2S,3R)-8-Bromo-3-hydroxy-2-methyl-2,3-dihydro-1H-pyrido[2,3-b][1,4]diazepin-4(5H)-one (compound 148) and (2R,3S)-8-bromo-3-hydroxy-2-methyl-2,3-dihydro-1H-pyrido[2,3-b][1,4]diazepin4(5H)-one (connection 149).

Enantiomers were separated by chiral preparative HPLC using a Chiralpak® IC (Daicel Ltd.) column (2x25 cm), flow rate 13.5 ml min ^-1 , eluting with a mixture of 10% ethanol in heptane + 0.2% diethylamine, UV determination at 254 nm . Samples are loaded onto the column through a vacuum pump built into the column, pumped with ethanol (1.5 ml min ^-1 ) throughout the entire run, obtaining a volume

- 88 046887 fixed flow rate 15 ml min ^-1 . Chirality is determined arbitrarily.

First enantiomer eluted (compound 148). Rt 9.6 min (method 4b). The second enantiomer eluted (compound 149). Rt 13.5 min (method 4b).

Step 5. (E)-3-((2R,3S)-3-Hydroxy-2-methyl-4-oxo-2,3,4,5-tetrahydro-1H-pyrido[2,3b][1,4 ]diazepin-8-yl)-N-methyl-N-((3-methylbenzofuran-2-yl)methyl)acrylamide (compound 150).

Namethyl-Na((3-methylbenzofuran-2-yl)methyl)acrylamide 9 (42 mg, 0.18 mmol), (2R,3S)-8-bromo-3-hydroxy-2-methyl- 2,3-dihydro-1H-pyrido[2,3-b][1,4]diazepin4(5H)-one 149 (50 mg, 0.18 mmol), NBu ₄ Cl (5.0 mg, 0.02 mmol) and [P(tBu) ₃ ]Pd(crotyl)Cl (Pd-162) (7.0 mg, 0.02 mmol) and the tube was evacuated and backfilled with N2 three times. 1,4-Dioxane (5.0 mL) and N-cyclohexyl-N-methylcyclohexanamine (79 μL, 0.37 mmol) were added and the reaction mixture was heated to 80°C and stirred for ~16 h. The reaction mixture was cooled to RT. the solvent is removed in vacuo and the solid is washed with isohexane. The crude product was then purified by column chromatography (0-3% MeOH/DCM) to obtain the desired product as a pale yellow solid. The solid is partially dissolved in MeCN and water is added until the product precipitates. The precipitate was collected by filtration and dried by azeotropic distillation with MeCN (2x2 ml) to obtain the desired product 150 as a pale yellow solid (35 mg, 45%). Rt 1.83 min (method 1a) m/z 421 [M+H]+ (ER+). 1H NMR (500 MHz, DMSO-66, 363 K): δ, ppm 9.80 (s, 1H), 7.98 (d, J=1.9 Hz, 1H), 7.59-7, 52 (m, 1H), 7.49-7.37 (m, 3H), 7.31-7.22 (m, 2H), 7.12 (d, J=15.7 Hz, 1H), 5 .91 (d, J=5.7 Hz, 1H), 4.84 (s, 2H), 4.76 (d, J=4.7 Hz, 1H), 4.22 (dd, J=4, 7 Hz, 3.4 Hz, 1H), 3.82-3.71 (m, 1H), 3.10 (s, 3H), 2.27 (s, 3H), 1.12 (d, J= 6.5 Hz, 3H).

Example 26. Synthesis of (E)-3-((2R,3S)-3-hydroxy-2-methyl-4-oxo-2,3,4,5-tetrahydro-1H-pyrido[2,3b][1, 4]diazepin-8-yl)-N-methyl-N-((2-methylbenzofuran-3-yl)methyl)acrylamide (compound 151).

General synthesis scheme.

149 151

Reaction conditions: a) Pd-162, MeNCy ₂ , NBu ₄ Cl, 1,4-dioxane, 80°C.

Step 1. (E)-3-((2R,3S)-3-Hydroxy-2-methyl-4-oxo-2,3,4,5-tetrahydro-1H-pyrido[2,3-b][1 ,4]diazepin-8-yl)-Nmethyl-^((2-methylbenzofuran-3-yl)methyl)acrylamide (compound 151).

Namethyl-N-((2-methylbenzofuran-3-yl)methyl)acrylamide 38 (42 mg, 0.18 mmol), (2R,3S)-8-bromo-3-hydroxy-2-methyl is loaded into the reaction tube -2,3-dihydro-1H-pyrido[2,3-b][1,4]diazepine4(5^-one 149 (50 mg, 0.18 mmol), NBu ₄ Cl (5.0 mg, 0. 02 mmol) and [P(tBu) ₃ ]Pd(crotyl)Cl (Pd-162) (7.0 mg, 0.02 mmol) and the tube is evacuated and backfilled with N ₂ three times. 1,4-dioxane is added. 5.0 mL) and N-cyclohexyl-N-methylcyclohexanamine (79 μL, 0.37 mmol), and the reaction mixture was heated to 80°C and stirred for 3 hours. The reaction mixture was cooled to RT, the solvent was removed in vacuo and the solid was washed. isohexane. The crude product is then purified by column chromatography (0-3% MeOH/DCM) to give the desired product as a pale yellow solid. The solid is partially dissolved in MeCN and water is added until the product precipitates. collected by filtration and dried by azeotropic distillation with MeCN (2x2 ml) to obtain the desired product 151 as a pale yellow solid (14 mg, 17%). Rt 1.80 min (method 1a) m/z 421 [M+H]+ (ER+). 1H NMR (500 MHz, DMSO-d^ 363 K): δ, ppm 9.80 (s, 1H), 7.98 (d, J=1.9 Hz, 1H), 7.54 (d, J=7.6 Hz, 1H), 7.49-7.42 (m, 2H), 7.39 (d, J=2.0 Hz, 1H), 7.26-7.15 (m, 2H ), 7.10 (d, J=15.9 Hz, 1H), 5.89 (d, J=5.8 Hz, 1H), 4.80-4.73 (m, 3H), 4.21 (dd, J=4.7 Hz, 3.4 Hz, 1H), 3.81-3.71 (m, 1H), 3.00 (s, 3H obscured by solvent peak), 2.50 (s, 3H , - darkened by the solvent peak), 1.11 (d, J=6.5 Hz, 3H).

Example 27. Synthesis of ^,E-N((7-(4-cyanophenoxy)-2-methylbenzofuran-3-yl)methyl)-3-(3-hydroxy-3methyl-4-oxo-2,3,4,5 -tetrahydro-1H-pyrido[2,3-b][1,4]diazepin-8-yl)-N-methylacrylamide (compound 152).

General synthesis scheme.

!

Reaction conditions: a) HOBt-^O, EDC-HCl, DIPEA, DMF.

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Step 1. ^E-N((7-(4-Cyanophenoxy)-2-methylbenzofuran-3-yl)methyl)-3-(3-hydroxy-3-methyl-4oxo-2,3,4,5-tetrahydro -1H-pyrido[2,3-b][1,4]diazepin-8-yl)-Nmethylacrylamide (compound 152) is prepared from amine 79 and acid 142 as a yellow solid in the standard amine-amide coupling reaction described , for example, for compound 143 (purity=95%; yield=38%). Rt 1.72 min; LCMS: m/z: 538.5 [M+H]+. ¹ H NMR (400 MHz, DMSO-06): δ, ppm 9.93 (s, 1H), 7.98 (s, 1H), 7.83 (d, J=8.9 Hz, 2H) , 7.57-7.19 (m, 4H), 7.15-6.99 (m, 4H), 6.24 (shs, 1H), 5.28 (shs, 1H), 4.91 (s , 0.3H, rotamer), 4.76 (s, 1.7H, rotamer), 3.22-3.09 (m, 2H), 3.07 (s, 2.3H, rotamer), 2.87 (s, 0.7H, rotamer), 2.48 (s, 3H, partly under the DMSO peak), 1.23 (s, 3H).

Example 28. Synthesis of (E)-N-((7-(3-methoxyphenoxy)-2-methylbenzofuran-3-yl)methyl)-N-methyl-3-(4oxo-2,3,4,5-tetrahydro- 1H-pyrido[2,3-b][1,4]diazepin-8-yl)acrylamide (compound 155).

General synthesis scheme.

Reaction conditions: a) N,N-dimethylglycine, CuI, K ₂ PO ₄ , DMSO, 100°C; b) 1-chloroethyl chloroformate, DCM, 0°C to RT; c) GATU, DIPEA, DMF.

No.(4-Methoxybenzyl)-1-(7-(3-methoxyphenoxy)-2-methylbenzofuran-3-yl)-No.

Stage

1.

methylmethanamine (compound 153).

Mixture of 1-(7-bromo-2-methylbenzofuran-3-yl)-N-(4-methoxybenzyl)-N-methylmethanamine 76 (200 mg, 0.53 mmol), 3-methoxyphenol 124 (Sigma-Aldrich) (80 mg, 0.64 mmol), 2-(dimethylamino)acetic acid (11 mg, 0.11 mmol), copper(1) iodide (10 mg, 0.05 mmol) and tribasic potassium phosphate (230 mg, 1.1 mmol) is evacuated and backfilled with nitrogen (3x). DMSO (2 ml) is added and the mixture is heated to 110°C overnight. Add additional aliquots of copper(I) iodide (10 mg, 0.05 mmol) and 3-methoxyphenol 124 (80 mg, 0.64 mmol) and heating is continued for a further 3 hours. The reaction mixture is cooled to rt, filtered through Celite® and the filtrate is diluted with brine (20 ml). The crude mixture is extracted with ethyl acetate (2x10 ml) and the combined organic phases are dried over Na ₂ SO ₄ and concentrated in vacuo. The crude product was purified by chromatography (0100% EtOAc/isohexane) to give the title compound 153 as a light orange oil (103 mg, 43%). R _t 1.76 min (method 1a) m/z 418 [M+H] ⁺ (ER ⁺ ).

Step 2. 1-(7-(3-Methoxyphenoxy)-2-methylbenzofuran-3-yl)-Nmethylmethanamine (compound 154).

To an ice-cold solution of No.(4-methoxybenzyl)-1-(7-(3-methoxyphenoxy)-2-methylbenzofuran-3-yl)-No. methylmethanamine 153 (100 mg, 0.24 mmol) in DCM (2 ml) add 1 -chloroethyl chloroformate (0.031 ml,

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0.29 mmol). The mixture is allowed to reach RT and stirred for a further 20 min, then evaporated to dryness. The residue was taken up in methanol (5 ml) and heated to reflux for 1 hour. The mixture was evaporated to dryness and the residue was dissolved in methanol (4 ml) and applied to an SCC column (2 g). The column was washed with methanol (20 ml) and the product was eluted with 10% methanolic ammonia (10 ml) to give the title compound 154 as a colorless gum (42 mg, 47%), which was used in the next step without further purification. Rt 1.40 min (method 1b) m/z 267 [M - NHCH ₃ ] ⁺ (er ⁺ ).

/

Ome

Step 3. (E)-N-((7-(3-Methoxyphenoxy)-2-methylbenzofuran-3-yl)methyl)-N-methyl-3-(4-oxo-2,3,4,5tetrahydro-1H -pyrido[2,3-b][1,4]diazepin-8-yl)acrylamide (compound 155).

To a solution of 1-(7-(3-methoxyphenoxy)-2-methylbenzofuran-3-yl)-K-methylmethanamine 154 (40 mg, 0.11 mmol), trifluoroacetate (E)-3-(4-oxo-2, 3,4,5-tetrahydro-1H-pyrido[2,3-b][1,4]diazepin-8yl)acrylic acid 6 (31 mg, 0.09 mmol) and DIPEA (78 μl, 0.45 mmol) in DMF (2 ml) add GATU (41 mg, 0.11 mmol) and the mixture is stirred at RT for 1 hour. The reaction mixture is quenched by adding water (10 ml) and the solid is collected and dried. The crude product was purified by chromatography (0-10% MeOH/DCM) to give the title compound 155 as a pale yellow solid (25 mg, 54%). R _t 2.17 min (method 1a) m/z 513 [M+H] ⁺ (ER ⁺ ). ¹ H NMR (500 MHz, DMSO-06, 363 K): δ, ppm 9.13 (s, 1H), 7.99 (d, J=2.0 Hz, 1H), 7.47 (d , J=15.5 Hz, 1H), 7.41-7.34 (m, 2H), 7.26 (t, J=8.2 Hz, 1H), 7.17 (t, J=7, 9 Hz, 1H), 7.10 (d, J=15.6 Hz, 1H), 6.88 (dd, J=7.9 Hz, 1.0 Hz, 1H), 6.72 (dd, J =8.3 Hz, 2.4 Hz, 0.8 Hz, 1H), 6.61 (t, J=2.4 Hz, 1H), 6.57 (ddd, J=8.1 Hz, 2, 3 Hz, 0.8 Hz, 1H), 5.80 (s, 1H), 4.78 (s, 2H), 3.76 (s, 3H), 3.47-3.42 (m, 2H) , 3.03 (s, 3H), 2.70-2.63 (m, 2H), 2.48 (s, 3H).

Example 29. Synthesis of (R)-3-hydroxy-3-methyl-8-((E)-3-oxo-3-((3aS,6aR)-5-(4-phenoxyphenyl)3,3a,4,6a -tetrahydrocyclopenta[c]pyrrol-2(1H)-yl)prop-1-en-1-yl)-1,2,3,5-tetrahydro-4H-pyrido[2,3^[M^iazepinA-one ( connection 159).

Reaction conditions: a) aq. 2M K2CO3, DME, Pd(PPh ₃ %; b) TFA/DCM, 0°C to RT; c) HOBT, EDC, DIPEA, DMF.

Step 1. tert-Butyl (3aS,6aR)-5-(4-phenoxyphenyl)-3,3a,4,6a-tetrahydrocyclopenta[c]pyrrole2(III)-carboxylate (compound 157) was synthesized according to the procedure described in US2017/ 0174683 A1. Add tert-butyl (3aR,6aR)-5-(((trifluoromethyl)sulfonyl)oxy)-3,3a,4,6atetrahydrocyclopenta^pyrrole^Nicarboxylate 57 (104 mg, 0.276 mmol), (4-phenoxyphenyl) to the microwave test tube )boronic acid 156 (96 mg, 0.448 mmol) and a solution of 2 M K ₂ CO ₃ (276 μl, 0.552 mmol) in ethylene glycol dimethyl ether (1 ml) were purged with nitrogen for 10 min in a microwave bath. Then

- 91 046887 add tetrakisphenylphosphine palladium (30 mg, 0.025 mmol). The resulting suspension is stirred at 100°C for 30 minutes under microwave irradiation. The resulting solution is cooled to RT. Add water and EtOAc, the organic layer is separated, washed with water, then brine and dried (MgSO ₄ ) and evaporated to dryness to give a brown oil, which is purified on silica gel using a SNAP ultra column (10 g) and n-heptane/AcOEt 95 /5 to 50/50 as eluent. The good fractions were collected and concentrated to dryness to give tert-butyl (3aR,6aS)-5-(4phenoxyphenyl)-3,3a,4,6a-tetrahydrocyclopenta[c]pyrrole-2(1H)-carboxylate 157 as a colorless oil, which crystallizes as a white solid on standing (85 mg, purity=99%, yield=85%). RT 2.65 min; LCMS: m/z: 322.2 [M+H]+.

Step 2. (3aS,6aR)-5-(4-Phenoxyphenyl)-1,2,3,3a,4,6a-hexahydrocyclopenta[c]pyrrole 2,2,2trifluoroacetate (compound 158).

To a solution of tert-butyl (3aR,6aS)-5-(4-phenoxyphenyl)-3,3a,6,6a-tetrahydro-1H-cyclopentaIpyrrole-2-carboxylate 157 (85 mg, 0.23 mmol) in DCM (1 .0 ml) add 2,2,2trifluoroacetic acid (25.67 mg, 0.23 mmol) dropwise. The reaction mixture is stirred at room temperature until complete conversion of the starting material is determined. After completion of the reaction, the solvents were removed in vacuo to give the title compound 158 (88 mg) as a yellow oil. Rt 1.29 min (method 1b); m/z 278.6 [M+H]+ (ER ⁺ ).

Step 3. (R)-3-Hydroxy-3-methyl-8-((E)-3-oxo-3-((3aS,6aR)-5-(4-phenoxyphenyl)-3,3a,4,6atetrahydrocyclopentai pyrrol-2(1I)-i.l)prop-1-en-1-yl)-1,2,3,5-tetrahydro-4H-pyrido[2,3-b][1,4]diazepine- 4-on (compound 159).

(3aR,6aS)-5-(4phenoxyphenyl)-1,2,3,3a,6,6a-hexahydrocyclopenta[c]pyrrole 2,2,2-trifluoroacetate 158 (44.6 mg) is added to a microwave test tube covered with aluminum foil ,

0.11 mmol), HOBt-H ₂ O (26.18 mg, 0.17 mmol, 1.5 eq.), EDC-HCl (32.77 mg, 0.17 mmol) and (E)-3- [(3R)-3hydroxy-3-methyl-4-oxo-2,5-dihydro-1H-pyrido[2,3-b][1,4]diazepin-8-yl]prop-2-enoic acid 142 ( 30 mg, 0.11 mmol), and the solid was dissolved in DMF (0.2 ml). After 2 min of stirring the QD, DIPEA (0.07 mL, 0.40 mmol) was added. The reaction mixture is stirred in the dark overnight at the same temperature. After completion of the reaction, water and acetonitrile are added to the mixture, the suspension is treated with ultrasound, the precipitate is filtered off and washed with MTBE. The crude product (18 mg) is further purified by autoprep. The title compound 159 (6.6 mg) was isolated as a yellow foam. Rt 1.91 min (method 1b); m/z 523.5 [M+H]+ (ER ⁺ ). ¹ H NMR (400 MHz, DMSO-dg): δ, ppm ¹ H NMR (400 MHz, DMSO-dg): δ, ppm 9.95 (d, J=5.3 Hz, 1H), 7.92 (m, 1H), 9.50 (d, J=8.3 Hz, 2H), 7.42-7.28 (m, 4H), 7.14 (t, J=6.2 Hz , 1H), 7.03-6.95 (m, 4H), (dd, J=6.8 Hz, 15.5 Hz, 1H), 6.28-6.21 (m, 1H), 6, 13 (shs, 1H), 5.30 (d, J=4.4 Hz, 1H), 4.05-3.75 (m, 2H), 3.73-3.49 (m, 2H), 3 .22-3.10 (m, 2H), 3.10-2.91 (m, 2H), 2.69-2.55 (m, 2H), 1.23 (d, J=4.1 Hz , 3H). Note: Aliphatic signals are partially obscured by water and solvent peaks.

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Example 30. Synthesis of (E)-N-((7-((2,3-dihydrobenzofuran-6-yl)oxy)-2-methylbenzofuran-3-yl)methyl)3-(2,2-dimethyl-3 - oxo-1,2,3,4-tetrahydropyrido[2,3-b]pyrazin-7-yl)-Umethylacrylamide (compound 164).

Reaction conditions: a) CuI, X,X-dimethylglycine, K ₂ PO ₄ , 1,4-dioxane, 100°C; b) Pd/C, MeOH, _H2 , 45°C; c) acryloyl chloride, TEA, DCM, 0°C to RT; d) Pd-116, DIPEA, Bu4NCl, 1,4-dioxane, 80°C.

Step 1. 1-(7-((2,3-Dihydrobenzofuran-6-yl)oxy)-2-methylbenzofuran-3-yl)-U(4-methoxybenzyl)N-methylmethanamine (compound 161).

Mixture of 3-(((4-methoxybenzyl)(methyl)amino)methyl)-2-methylbenzofuran-7-ol 76 (200 mg,

0.64 mmol), 6-bromo-2,3-dihydrobenzofuran 160 (153 mg, 0.77 mmol), yN-dimethylglycine (66 mg, 0.64 mmol), CuI (122 mg, 0.64 mmol) and K3PO4 (273 mg, 1.3 mmol) was evacuated and backfilled with nitrogen three times. DMSO (3 ml) was added and the mixture was heated to 110°C and stirred for 16 hours. The reaction mixture was cooled to RT. Ethyl acetate (10 ml) was added and the mixture was filtered through Celite®. The resulting solution was washed with water (10 ml) and brine (10 ml), dried over MgSO ₄ , filtered and concentrated in vacuo. The crude product was purified by column chromatography (0-10% MeOH/DCM) to give the title compound 161 as a colorless oil (80 mg, 27%). Rt 1.77 min (method 1a) m/z 430 [M+H]+ (ER ⁺ ).

Step 2. 1-(7-((2,3-Dihydrobenzofuran-6-yl)oxy)-2-methylbenzofuran-3-yl)-N-methylmethanamine (compound 162).

Solution of 1-(7-((2,3-dihydrobenzofuran-6-yl)oxy)-2-methylbenzofuran-3-yl)-N-(4-methoxybenzyl)N-methylmethanamine 161 (80 mg, 0.19 mmol) in methanol (5 ml) is passed through an H-cube with a 10% Pd/C catalytic cartridge and using the 'Full H ₂ ' mode at 45°C. The solvent was concentrated in vacuo to give the title compound 162 as a colorless oil (43 mg, 70%). Rt 1.45 min (method 1a) m/z 279 [M - MeNH]+ (ER ⁺ ).

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Step 3. ^((7-((2,3-Dihydrobenzofuran-6-yl)oxy)-2-methylbenzofuran-3-yl)methyl)-N methylacrylamide (compound 163).

To an ice-cold solution of 1-(7-((2,3-dihydrobenzofuran-6-yl)oxy)-2-methylbenzofuran-3-yl)-No. methylmethanamine 160 (160 mg, 0.52 mmol) and TEA (300 μl, 2.07 mmol) in DCM (10 ml) add dropwise a solution of acryloyl chloride (42 µl, 0.52 mmol) in DCM (1.0 ml). The reaction mixture is warmed to RT and stirred for 1 hour. The reaction mixture is quenched with water (5.0 ml) and the two phases are separated. The organic phase is dried with MgSO ₄ , filtered and concentrated in vacuo. The crude product was purified by chromatography (0-100% EtOAc/isohexane) to give the title compound 163 as a colorless oil (120 mg, 60%). R _t 2.36 min (method 1a); m/z 364 [M+H]+ (ER ⁺ ).

164

Step 4. (E)-N-((7-((2,3-Dihydrobenzofuran-6-yl)oxy)-2-methylbenzofuran-3-yl)methyl)-3-(2,2dimethyl-3-oxo- 1,2,3,4-tetrahydropyrido[2,3-b]pyrazin-7-yl)-Nmethylacrylamide (compound 164).

^((7-((2,3-dihydrobenzofuran-6-yl)oxy)-2methylbenzofuran-3-yl)methyl)-^methylacrylamide 163 (60 mg, 0.16 mmol), 7-bromo- 2,2-dimethyl-1,2dihydropyrido[2,3-b]pyrazin-3(4H)-one 14 (42 mg, 0.16 mmol), tetrabutylammonium chloride hydrate (5.0 mg, 0.02 mmol) and Pd[P(tBu) ₃ ] ₂ (Pd-116) (9.0 mg, 0.02 mmol) and the tube was evacuated and backfilled with N2 three times. 1,4-dioxane (4.0 ml) and DIPEA (58 μl, 0.33 mmol) were added and the reaction mixture was heated to 80°C and stirred overnight. The reaction mixture is cooled to RT, and the solvent is removed in vacuo. The crude product was then purified by column chromatography (0-3% MeOH/DCM) to give the desired product 164 as a pale yellow solid (66 mg, 74%). Rt 2.72 min (method 1b); m/z 539 [M+H] ⁺ (ER ⁺ ). ^1H NMR (500 MHz, DMSO-dg, VT NMR): δ, ppm 10.42 (s, 1H), 7.89 (d, J=1.9 Hz, 1H), 7.48 (d , J=15.4 Hz, 1H), 7.34 (d, J=7.8 Hz, 1H), 7.25 (d, J=1.9 Hz, 1H), 7.18-7.13 (m, 2H), 7.04 (d, J=15.4 Hz, 1H), 6.83 (dd, J=7.9 Hz, 1.0 Hz, 1H), 6.47 (dd, J =8.1 Hz, 2.3 Hz, 1H), 6.42 (d, J=2.2 Hz, 1H), 6.04 (s, 1H), 4.78 (s, 2H), 4, 57 (t, J=8.6 Hz, 2H), 3.15 (t, J=8.6 Hz, 2H), 3.03 (s, 3H), 2.49 (s, 3H), 1, 31 (s, 6H).

Example 31. Synthesis of ^,E-^((7-amino-2-methylbenzofuran-3-yl)methyl)-3-(3-hydroxy-3-methyl-4oxo-2,3,4,5-tetrahydro-1H -pyrido[2,3-b][1,4]diazepin-8-yl)-N-methylacrylamide (compound 167).

General synthesis scheme.

Reaction conditions: a) Pd ₂ (dba) ₃ , Xantphos, benzophenonymine 72, Cs ₂ CO ₃ , PhCh ₃ , 110°C; b) i) TPA, DXM; ii) acryloyl chloride, TEA, DCM, 0°C to RT; c) i) Pd-116, NBu4Cl, DIPEA, 1,4-dioxane, 80°C; ii) 1M HCl, DCM.

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Vos

Vg

Benzophenoneimine 72, Pd ₂ (dba) ₃ ,

XantPhos, Cs ₂ CO ₃ , PhCH ₃ , 110 °C

Ph

Weight

Ph

Step 1. tert-Butyl ((7-((diphenylmethylene)amino)-2-methylbenzofuran-3-yl)methyl)(methyl)carbamate (compound 165).

tert-Butyl ((7-bromo-2-methylbenzofuran-3-yl)methyl)(methyl)carbamate 97 (2.0 g, 5.65 mmol), tris(dibenzylideneacetone) dipalladium (0.52 g, 0.57 mmol), Cs ₂ CO ₃ (3.68 g, 11.3 mmol) and Xantphos (0.49 g, 0.85 mmol) were added to the flask and the flask was evacuated and backfilled with N2 three times. Add toluene (40 ml) and N2 is bubbled through the reaction mixture for 10 minutes. Benzophenonymine 72 (1.1 mL, 6.78 mmol) was then added and the reaction mixture was heated to reflux for ~24 h. The reaction mixture was cooled to RT, stirred over the weekend and then concentrated in vacuo. The crude product was purified by column chromatography (DCM) to give the desired product 165 as a yellow oil that crystallized on standing (1.67 g, 65%). R 3.09 min (method 1b); m/z 455 [M+H]+ (ER ⁺ ). ^1H NMR (500 MHz, DMSO-dQ: δ, ppm 7.76-7.65 (m, 2H), 7.63-7.55 (m, 1H), 7.55-7.46 ( m, 2H), 7.33-7.21 (m, 3H), 7.16-7.04 (m, 3H), 7.01-6.88 (m, 1H), 6.49 (d, J=7.6 Hz, 1H), 4.40 (s, 2H), 2.61 (s, 3H), 2.37 (s, 3H), 1.43 (s, 9H).

Step 2. N((7-((Diphenylmethylene)amino)-2-methylbenzofuran-3-yl)methyl)-N~-methylacrylamide (compound 166).

TFA (15 ml, 195 mmol) is added dropwise to a stirred solution of tert-butyl ((7((diphenylmethylene)amino)-2-methylbenzofuran-3-yl)methyl)(methyl)carbamate 165 (0.8 g, 1. 76 mmol) in DCM (30 ml) and the reaction mixture was stirred at RT for 1 hour. The reaction mixture was concentrated in vacuo and the resulting residue was taken up in DCM (30 ml), cooled to 0°C and added TEA (5.0 ml , 35.9 mmol), then acryloyl chloride (0.17 ml, 2.11 mmol). The reaction mixture is returned to RT and stirred for ~16 hours. The reaction mixture is quenched with water (50 ml), then the organic phase is separated and the aqueous phase is again extracted with DCM (2x50 ml). The combined organic layers are dried by passing through a phase separator cartridge and concentrated in vacuo. The crude product was purified by column chromatography (0-3% MeOH/DCM) to give the crude product, which was again purified by column chromatography (0-30% EtOAc/isohexane) to give the desired product 166 as a yellow oil (0.16 g, 21 % for 2 stages).

Ph

i) Pd-116, DIPEA, NBu ₄ CI,

1,4-dioxane, 80 °C;

ii) 1M HCL DXM

Step 3. (R,E-N-((7 -Amino-2-methylbenzofuran-3 -yl)methyl)-3 -(3 -hydroxy-3 -methyl-4-oxo-2,3,4,5tetrahydro-1H -pyrido[2, 3-b][1,4]diazepin-8-yl)^-methylacrylamide (compound 167).

Na((7-((diphenylmethylene)amino)-2-methylbenzofuran-3yl)methyl)-N~-methylacrylamide 166 (0.12 g, 0.29 mmol), ^)-8-bromo-3 is loaded into the reaction tube -hydroxy-3-methyl-2,3-dihydro-Spirido[2,3-b][1,4]diazepin-4(5H)-one 47 (80 mg, 0.29 mmol), NBu ₄ Cl (9 mg, 0.03 mmol) and Pd-116 (15 mg, 0.03 mmol) and the tube is evacuated and backfilled with N ₂ three times. Add 1,4-dioxane (5 ml) and DIPEA (0.10 ml, 0.59 mmol) and the reaction mixture is heated to 80°C and stirred for 3 hours. The reaction mixture is cooled to RT, then diluted with HCl (10 ml, 1 M aq) and the aqueous mixture was extracted with DCM (3x20 ml). The combined organic extracts are washed with aq. 1 M HCl (2x20 ml), then the combined aqueous layers are neutralized with sat. aq. _NaHCO3 . The resulting precipitate was collected by filtration and purified by column chromatography (0-5% MeOH/DCM) to obtain the desired product.

- 95 046887 duct 167 as a pale yellow solid (42 mg, 33% in 2 stages). R _t 1.32 min (method 1a); m/z 436 [M+H]+ (ER ⁺ ). ^1H NMR (500 MHz, DMSO-dg, 363 K): δ, ppm 9.36 (s, 1H), 7.94 (d, J=1.8 Hz, 1H), 7.44 (d , J=15.4 Hz, 1H), 7.36 (d, J=2.0 Hz, 1H), 7.08 (d, J=15.3 Hz, 1H), 6.87 (t, J =7.7 Hz, 1H), 6.74 (d, J=7.7 Hz, 1H), 6.53 (d, J=7.6 Hz, 1H), 6.03 (t, J=4 .6 Hz, 1H), 4.91 (s, 1H), 4.87 (s, 2H), 4.71 (s, 2H), 3.21 (dd, J=13.4, 3.7 Hz , 1H), 3.16 (dd, J=13.4, 5.6 Hz, 1H), 2.98 (s, 3H), 2.47 (s, 3H), 1.28 (s, 3H) .

Example 32. Synthesis of (E)-N-((7-amino-2-methylbenzofuran-3-yl)methyl)-N-methyl-3-(8-oxo-6,7,8,9tetrahydro-5H-pyrido[ 2,3-b]azepin-3-yl)acrylamide (compound 168).

General synthesis scheme.

Ph

Reaction conditions: a) i) TPA, DCM; ii) GATU, DIPEA, DMF, iii) 1M HCl, DCM.

Step 1. (E)-N-((7-Amino-2-methylbenzofuran-3-yl)methyl)-N-methyl-3-(8-oxo-6,7,8,9-tetrahydro5H-pyrido[2 ,3-b]azepin-3-yl)acrylamide (compound 168).

TFA (2.5 mL, 32.4 mmol) was added dropwise to a stirred solution of tert-butyl ((7((diphenylmethylene)amino)-2-methylbenzofuran-3-yl)methyl)(methyl)carbamate 165 (0.15 g, 0.21 mmol) in DCM (5.0 ml) and the reaction mixture was stirred at RT for 1 hour. The reaction mixture was concentrated in vacuo and the resulting residue was taken up in DMF (2.0 ml) and trifluoroacetate (E)- 3(8-oxo-6,7,8,9-tetrahydro-5H-pyrido[2,3-b]azepin-3-yl)acrylic acid 4 (75 mg, 0.21 mmol), then DIPEA (1, 0 ml, 5.73 mmol). The reaction mixture was stirred for 10 min, then GATU (0.12 g, 0.32 mmol) was added and the reaction mixture was stirred for an additional ~16 h at RT. The reaction mixture is diluted with aq. 1 M HCl (10 ml) and extract with DCM (3x20 ml). The combined organic extracts are washed with HCl (2x20 ml, 1 M aq) and the combined aqueous layers are neutralized with sat. aq. _NaHCO3 . The resulting precipitate was collected by filtration and purified by column chromatography (05% MeOH/DCM) to give the desired product 168 as an off-white solid (40 mg, 43% in 3 steps). R _t 1.41 min (method 1a); m/z 405 [M+H]+ (ER+). ^1H NMR (500 MHz, DMSO-dfo 363 K): δ, ppm 9.55 (s, 1H), 8.47 (d, J=2.2 Hz, 1H), 8.02 (d, J=2.2 Hz, 1H), 7.54 (d, J=15.4 Hz, 1H), 7.34-7.20 (m, 1H), 6.87 (t, J=7.6 Hz, 1H), 6.75 (d, J=7.7 Hz, 1H), 6.53 (dd, J=7.6 Hz, 1.1 Hz, 1H), 4.86 (s, 2H) , 4.73 (s, 2H), 2.97 (s, 3H), 2.76 (t, J=7.2 Hz, 2H), 2.47 (s, 3H), 2.31 (t, J=7.2 Hz, 2H), 2.14 (p, J=7.2 Hz, 2H).

Example 33. Synthesis of (E)-N-methyl-N-((2-methylbenzofuran-3-yl)methyl)-3-(4-oxo-2,3,4,5-tetrahydro1H-pyrido[2,3- b][1,4]diazepin-8-yl)acrylamide (compound 169).

General synthesis scheme.

Reaction conditions: a) Pd-162, NCy ₂ NMe, NBu ₄ Cl, 1,4-dioxane, 80°C 8-bromo-1,2,3,5-tetrahydro-4Hpyrido[2,3-b][1 ,4]diazepin-4-one (compound 169) is synthesized according to methods described in the literature, for example, AFFINIUM PHARMECEUTICALS, INC: WO2007/67416, 2007, A2 and AURIGENE DISCOVERY TECHNOLOGIES LIMITED; WO2013/80222, 2013, A1.

Step 1. (E)-N-Methyl-N-((2-methylbenzofuran-3-yl)methyl)-3-(4-oxo-2,3,4,5-tetrahydro-1Hpyrido[2,3-b ][1,4]diazepin-8-yl)acrylamide (compound 170).

Namethyl-U-((2-methylbenzofuran-3-yl)methyl)acrylamide 38 (150 mg, 0.65 mmol), 8-bromo-2,3-dihydro-1H-pyrido[2,3] is loaded into the reaction tube -b][1,4]diazepin-4(5H)-one 169 (158 mg, 0.65 mmol), tetrabutylammonium chloride hydrate (19.36 mg, 0.07 mmol), [P(tBu) ₃ ]Pd (crotyl)Cl (Pd-162) (26.1 mg, 0.07 mmol). The reaction tube is flushed with nitrogen for 5 minutes. 1,4-Dioxane (10 mL) and N-cyclohexyl-N-methylcyclohexanamine (280 μL, 1.31 mmol) were added and the reaction mixture was purged with nitrogen for a further 5 min. The mixture is heated to 80°C for 1 hour. The reaction mixture is cooled to RT and the solid is collected by filtration, washed with 1,4-dioxane (10 ml), then acetonitrile (10 ml), and the solid is dried in vacuum to obtain the above title compound 170 as a pale yellow solid (192 mg, 74%). Rt 1.83 min (method 1b) m/z 391 [M+H]+ (ER+). ¹ H NMR (400 MHz, D\1С(.)-с1 ₆ , 363 K): δ, ppm 9.17 (s, 1H), 7.98 (d, J=2.0 Hz, 1H) , 7.54 (d, J=7.5 Hz, 1H), 7.50-7.43 (m, 2H), 7.38 (d, J=2.0 Hz, 1H), 7.26- 7.16 (m, 2H), 7.10 (d, J=15.5 Hz, 1H), 5.81 (d, J=4.8 Hz, 1H), 4.77 (s, 2H), 3.47-3.41 (m, 2H), 2.69-2.62 (m, 2H). Note: Both CH ₃ signals are obscured by the water and solvent peaks.

- 96 046887

Example 34. Synthesis of 2-hydroxyethane-1-aminium phosphate (E)-(2,2-dimethyl-7-(3-(methyl((3methylbenzofuran-2-yl)methyl)amino)-3-oxoprop-1-ene -1-yl)-3-oxo-2,3-dihydropyrido[2,3-b]pyrazin4(1H)-yl)methyl (compound 172).

General synthesis scheme.

Reaction conditions: a) 1) 18-crown-6, NMP/N-ethylpyrrolidone, RT 2) t-BuOK at 10°C, then chloromethyl bis-[2-(trimethylsilyl)ethyl]phosphate 21, 24 h, RT ; b) TPA/DXM, long-term processing

1. 18-crown-6, NMP/N-ztylpyrrolidone, CT 2 t-BuOK at 10 °C then chloromethyl bis[2-(trimethylsilyl)ethyl]phosphate 21, 24 h, CT

Stage 1. bis-(2-(Trimethylsilyl)ethyl)phosphate (E)-(3,3-dimethyl-6-(3-(methyl((3-methylbenzofuran-2yl)methyl)amino)-3-oxoprop-1 -en-1-yl)-2-oxo-3,4-dihydroquinoxalin-1 (2H)-yl)methyl (compound

171) .

In a hermetically sealed 6 ml microwave tube under N2, (E)-3-(2,2-dimethyl-3-oxo-1,4dihydropyrido[2,3-b]pyrazin-7-yl)-N-methyl-N- [(3-methylbenzofuran-2-yl)methyl]prop-2-enamide 19 (500 mg, 1.23 mmol, 1.00 eq.) and 1,4,7,10,13,16-hexaochacyclooctadecane (408, 37 mg, 1.53 mmol, 1.24 eq.) were placed in suspension in N-ethylpyrrolidone (3.0 ml) at RT. The resulting yellow suspension is then stirred at RT for 5 min, and degassing cycles are carried out (5 cycles; vacuum/nitrogen). The reaction mixture is then cooled to 10°C and potassium 2-methylpropan-2-ol (1.3 mL, 1.3 mmol, 1.05 eq, 1 M in THF) is added dropwise over 5 min. The resulting yellow solution is again subjected to degassing cycles (3 cycles; vacuum/nitrogen) and then stirred at RT for 20 min. Chloromethyl bis-(2-(trimethylsilyl)ethyl)phosphate 21 (1997.1 mg, 1.73 mmol, 1.4 eq., 30 w/w% in heptane) was then added dropwise to the reaction mixture at RT. for 5 minutes The reaction mixture was then stirred at RT for 22 hours (UPLC/MS: Starting material: R _t =1.64 min; 41% (IQ); ER ⁺ : 405.4. Product: R _t =2.68 min; 44 % (PIK); ER ⁺ : 715.6). A mixture of saturated ammonium acetate solution (7 ml; pH 9) and THF (5 ml) was added to the reaction mixture. After shaking, the phases are separated and a second wash is carried out with a mixture of brine (7 ml) and THF (5 ml). A third wash is carried out with brine (7 ml) and methylcyclohexane (7 ml), then added to the organic phase. The resulting suspension was filtered through a glass filter and the solid was then washed with methylcyclohexane (7 ml). The filtrate is concentrated to dryness at 25°C under vacuum from to

- 97 046887 by irradiation of a yellow oil, which is purified by flash chromatography (SiO ₂ ), eluting first with MTBE 99.8% DIEA 0.2%, then with MTBE 89.8%-Acetonitrile 10%-DIEA 0.2%. Fractions of interest were combined and concentrated to dryness at RT under vacuum to obtain the title compound 171 as a yellow oil (m=440.6 mg; UPLC purity: 98% (IIC); ES ⁺ : 715.6; 48. 9% yield).

Stage 2. 2-hydroxyethane-1-aminium phosphate (E)-(3,3-dimethyl-6-(3-(methyl((3-methylbenzofuran-2yl)methyl)amino)-3-oxoprop-1-ene- 1 -yl)-2-oxo-3,4-dihydroquinoxalin-1 (2H)-yl)methyl (compound

172) .

In a 50 ml flask containing bis-(2-trimethylsilylethyl)phosphate[2,2-dimethyl-7-[(E)-3-[methyl-[(3methylbenzofuran-2-yl)methyl]amino]-3-oxo -prop-1-enyl]-3-oxo-1H-pyrido[2,3-b]pyrazin-4-yl]methyl (440.6 mg, 0.60 mmol, 1 eq.) under N2 add anhydrous toluene ( 4 ml). The reaction mixture was stirred at 0°C until completely dissolved, and a solution of TFA (0.93 mL, 12.08 mmol, 20 eq.) in anhydrous DCM (2 mL) at 0°C was added dropwise over 1 min with stirring. The reaction mixture is then stirred at 0°C for a maximum of 1 min, and then immediately concentrated under high vacuum in a rotary evaporator (bath temperature: 1°C) until 2 ml remains. Anhydrous toluene (4 ml) was then added and the reaction mixture was concentrated until 2 ml remained. This pairing is repeated 4 times. A second co-evaporation cycle to dryness was then carried out twice, each time using anhydrous toluene (4 ml). The final residue is obtained as a thick yellow oil. Completion of hydrolysis is confirmed by UPLC/MS (product: _Rt 1.40 min; ER ^- : 513.5. Starting material: Rt 2.68 min; 0% (IIC)). Store in dry ice while preparing for the next step. In a 50 mL flask, prepare a solution of ethanolamine (186.31 mg, 3.02 mmol, 5 eq.) in acetone (10 mL) at 0°C, then add in one portion to a solution of the previous intermediate in dichloromethane (5 mL ) at 0°C. Immediate precipitation is observed and the resulting milky yellow suspension is stirred at 0°C for 10 minutes. After filtration through a glass filter, the sticky solid is washed with acetone (1x5 ml; 2x10 ml), then diethyl ether (2x10 ml). The suspension does not need to be filtered completely, and each time a layer of solvent must be present before proceeding to the next washing step, except for the last one. The resulting fine creamy solid is then sucked dry in only 2-3 seconds and final drying is carried out under vacuum at RT to obtain the title compound 172 as a yellow powder (m=253.7 mg; UPLC purity: 99% (IIC) ); ER ^- 513.5; 2 eq. ethanolamine; 1.3 wt.% TFA salt of ethanolamine; 1H NMR (400 MHz, D2O): δ, ppm 7.72 (d, J=3.8 Hz, 1H), 7.27-6.92 (m, 7H), 6.65 (d, J =15.6 Hz, 1H), 5.58 (dd, J=3.3 Hz, 11.2 Hz, 2H), 4.55 (s, 1H, rotamer), 4.49 (s, 1H, rotamer ), 3.74-3.70 (m, 4H), 3.06-3.02 (m, 4H), 2.90 (s, 2H, rotamer), 2.84 (s, 1H, rotamer), 1.98 (s, 2H, rotamer), 1.96 (s, 1H, rotamer), 1.26-1.20 (m, 6H).

Irimer 35. Synthesis of 2-hydroxyethane-1-aminium phosphate (E)-(8-(3-(((7-(3-methoxyphenoxy)-2methylbenzofuran-3-yl)methyl)(methyl)amino)-3-oxoprop - 1-en-1-yl)-4-oxo-1,2,3,4-tetrahydro-5H-pyrido[2,3-b][1,4]diazepin-5-yl)methyl (compound 174) .

General synthesis scheme.

/

St

- 98 046887

Reaction conditions: a) 1. 1 M t-BuOK/THF, DMSO, RT, 2. chloromethyl (trimethylsilyl)ethyl]phosphate 21; b) TPA/DCM, long-term processing.

bis-[2-

155

1. 1 M t-BuOK/THF, dmso, ct

2. chloromethyl bis[(trimethylsilyl)stype]phosphate 21

Step 1. bis-(2-(Trimethylsilyl)ethyl)phosphate (E)-(8-(3-(((7-(3-methoxyphenoxy)-2methylbenzofuran-3-yl)methyl)(methyl)amino)-3 -oxoprop-1-en-1-yl)-4-oxo-1,2,3,4-tetrahydro-5H-pyrido[2,3-b][1,4]diazepin-5-yl)methyl (compound 173).

In a sealed 6 ml microwave tube under nitrogen containing (E)-N-((7-(3methoxyphenoxy)-2-methylbenzofuran-3-yl)methyl)-N-methyl-3-(4-oxo-2,3 ,4,5-tetrahydro-1H-pyrido[2,3b][1,4]diazepin-8-yl)acrylamide 155 (400 mg, 0.76 mmol, 1.00 eq.) in suspension in anhydrous DMSO (3 .2 mL) at RT, add potassium 2-methylpropan-2-olate (0.83 mL, 0.83 mmol, 1.10 eq; 1 M in THF) dropwise over 2 min. The resulting dark yellow solution is then subjected to degassing cycles (5 cycles; vacuum/nitrogen) and then stirring at RT for 20 min. The reaction mixture was then added dropwise over 13 min to a solution of chloromethyl bis-(2-(trimethylsilyl)ethyl)phosphate 21 (1.47 mL, 1.06 mmol, 1.4 eq; 30% in hexane) with vigorous stirring. the reaction mixture (orange biphasic solution) is then stirred vigorously for 45 min to show good UPLC/MS conversion (starting material: R _t 1.82 min; 14% (AUC at 254 nm); ER ⁺ : 513.6). Product: Rt 2.66 min; 65% (AUC at 254 nm); ER ⁺ : 823.8). The reaction mixture is quenched with sat. AcONH4 solution (6 ml) and extracted four times with MTBE (4x6 ml). The combined organic phases are washed with brine and dried over Na2SO ₄ to obtain, after concentration under vacuum at 20°C, a thick orange oil (m=643 mg; UPLC/MS: 72% (AUC at 254 nm)).

Purification is carried out by flash chromatography (SiO ₂ ; eluting first with MTBE 99.5%-DIEA 0.5%, then with MTBE 93%-acetonitrile 6.5%-DIEA 0.5% and finally with MTBE 85%-acetonitrile 14, 5%-DIEA 0.5%). Fractions of interest were combined and concentrated to dryness at RT under vacuum to obtain the title compound as a yellow foam (m=219.5 mg; UPLC purity: 96% (AUC at 254 nm); 33.82% yield). The average yield was attributed to some degradation during flash chromatography (confirmed by 2D TLC experiments).

- 99 046887 /=L Jn ___ ft \ TPA/DXM, long-term treatment '°L Ф ° ¹⁷³ '^0

O V

Si

174 o

p'^O

O V

BUT. O nh ₃ BUT. /X

NH ₃ '

Stage 2. 2-hydroxyethane-1-aminium phosphate (E)-(8-(3-(((7-(3-methoxyphenoxy)-2-methylbenzofuran3-yl)methyl)(methyl)amino)-3-oxoprop- 1-en-1-yl)-4-oxo-1,2,3,4-tetrahydro-5H-pyrido[2,3b][1,4]diazepin-5-yl)methyl (compound 174).

Into a 100 ml flask containing bis-(2-trimethylsilylethyl)phosphate [8-[(E)-3-[[7-(3-methoxyphenoxy)-2methylbenzofuran-3 -yl]methylmethylamino]-3-oxoprop-1 - enyl]-4-oxo-2,3-dihydro-1H-pyrido[2,3-b][1,4]diazepin-5-yl]methyl 173 (219.5 mg, 0.26 mmol, 1, 0 eq.) under N ₂ add anhydrous toluene (4 ml). The resulting mixture was then stirred at 0°C until completely dissolved and a solution of 2,2,2-trifluoroacetic acid (0.93 ml, 12.08 mmol, 20 eq.) in anhydrous DCM (2.0 ml) at 0°C was added drop by drop for 30 s. The reaction mixture is stirred at 0°C for 3 min, and the reaction mass is then immediately concentrated under high vacuum with a rotary evaporator (bath temperature: 1°C) until 2 ml remains. Anhydrous toluene (4 ml) was then added and the reaction mixture was concentrated until 2 ml remained. This co-evaporation is repeated four times to remove all residual TPA (yellow foam). UPLC/MS (expected product R _t 1.58 min; ER: 621.6; 92% (IIC at 254 nm). API: R _t 1.81 min; 2% (IIC at 254 nm)). The original material does not remain.

In a 50 mL flask under _N2, prepare a solution of ethanolamine (186.31 mg, 3.02 mmol, 5.0 eq.) in THF (10 mL) at 0°C, then add one portion to a solution of the previous intermediate in THF (5 ml) at 0°C. Precipitation is observed after a few minutes of stirring at 0-5°C and the suspension is then filtered through a glass filter (the solid is clearly hygroscopic and filtration is stopped before the cake becomes dry). The thick suspension is washed with THF (2x10 ml; filtration is stopped each time just before completion). UPLC/MS of the filtrate showed the API as a smaller product and only traces of the expected prodrug). Finally the thick suspension is washed with diethyl ether (2 x 4 ml) and the solid is sucked dry within just a few seconds. The resulting yellow solid was then dried under vacuum at RT overnight to give the title compound (135.37 mg; 69.2% yield). UPLC/MS; method: 3 min acid_standard; 254 nm; purity 97.50% (max IIC graph), ER-: 621.58. 1H NMR (400 MHz, D2O, 333 K): δ, ppm 78.62 (s, 1H), 7.92 (brs, 1H), 7.85 (d, J=15.7 Hz, 1H) , 7.55 (m, 1H), 7.31 (m, 3H), 6.94 (m, 1H), 6.84 (m, 1H), 6.75 (s, 1H), 6.66 ( d, J=7.6 Hz, 1H), 5.97 (s, 1H, rotamer), 5.96 (s, 1H, rotamer), 5.10 (s, 1H, rotamer), 5.01 (s , 1H, rotamer), 4.22 (t, J=5.3 Hz, 4H), 4.03 (m, 2H), 3.91 (s, 1H, rotamer), 3.89 (s, 2H, rotamer), 3.54 (t, J=5.2 Hz, 4H), 3.31 (s, 3H), 2.88 (m, 1H), 2.78 (m, 1H), 2.64 ( s, 3H). Content of TPA-ethanolamine salt: 7.6 wt./wt.%.

- 100 046887

Example 36. Synthesis of ^,E-X-((7-(benzo^][1,3]dioxol-5-yloxy)-2-methylbenzofuran-3-yl)methyl)-3(3-hydroxy-3-methyl) -4-oxo-2,3,4,5-tetrahydro-III-pyrido[2,3-b][1,4]diazepin-8-yl)-X-methylacrylamide (compound 176).

General synthesis scheme.

176

Reaction conditions: a) acryloyl chloride, TEA, 0°C to RT; b) Pd-162, MeNCy ₂ , NBu ₄ Cl, 1,4-dioxane, 80°C.

Step 1. N((7-(Benzo^][1,3]dioxol-5-yloxy)-2-methylbenzofuran-3-yl)methyl)-X-methylacrylamide (compound 175).

To an ice-cold solution of 1-(7-(benzo^][1,3]dioxol-5-yloxy)-2-methylbenzofuran-3-yl)-No. methylmethanamine 118 (196 mg, 0.63 mmol) and TEA (350 μl , 2.52 mmol) in DCM (10 ml) add dropwise a solution of acryloyl chloride (51 μl, 0.63 mmol) in DCM (1.0 ml). After addition is complete, the mixture is heated to RT. The mixture is washed with water (5.0 ml) and the organics are separated through a phase separation cartridge and evaporated to dryness. The crude product was purified by column chromatography (0-50% EtOAc/isohexane) to give the title compound 175 as a clear, colorless oil (120 mg, 51%). Rt 2.29 min (method 1a) m/z 366 [M+H]+ (ER ⁺ ). ¹ H NMR (500 MHz, DMSO-06, 363 K): δ, ppm 7.27 (d, J=7.7 Hz, 1H), 7.13 (td, J=7.9, 0, 9 Hz, 1H), 6.90-6.76 (m, 3H), 6.70 (dd, J=2.6 Hz, 0.9 Hz, 1H), 6.51 (dd, J=8, 4 Hz, 2.4 Hz, 0.9 Hz, 1H), 6.18 (dd, J=16.7 Hz, 2.4 Hz, 0.9 Hz, 1H), 6.02 (d, J= 0.9 Hz, 2H), 5.71 (dd, J=10.5 Hz, 2.4 Hz, 1H), 4.71 (s, 2H), 2.94 (s, 3H), 2.47 (c, 3H).

176

Stage 2. ^,E-N((7-(Benzo^][1,3]dioxol-5-yloxy)-2-methylbenzofuran-3-yl)methyl)-3-(3hydroxy-3-methyl-4- oxo-2,3,4,5-tetrahydro-1H-pyrido[2,3-b][1,4]diazepin-8-yl)-N-methylacrylamide (compound 176).

Na((7-(benzo^][1,3]dioxol-5-yloxy)-2-methylbenzofuran-3yl)methyl)-X-methylacrylamide 175 (50 mg, 0.14 mmol), ^) is loaded into the reaction tube -8-bromo-3-hydroxy-3-methyl-2,3-dihydro-Spirido[2,3-b][1,4]diazepin-4(5H)-one 47 (37 mg, 0.14 mmol) , NBu4Cl (4.0 mg, 0.01 mmol) and

- 101 046887

[P(tBu)3]Pd(kpoTun)Cl (Pd-162) (6.0 mg, 0.01 mmol) and the tube was washed with N ₂ (5 min). Add 1,4dioxane (2.0 ml) and M-cyclohexyl-M-methylcyclohexanamine (59 µl, 0.27 mmol) and the reaction mixture is again purged with N2 (5 min). The reaction mixture is heated to 80°C for 1 hour, then cooled to RT and evaporated to dryness. The residue was triturated with isohexane (10 mL) and the crude solid was purified by chromatography (0-5% MeOH/DCM) to give the title compound 176 as a pale yellow solid (41 mg, 53%). R _t 2.15 min (method 1a); m/z 557 [M+H]+ (ER ⁺ ). 1H NMR (500 MHz, DMSODb, 363 K): δ, ppm 9.37 (s, 1H), 7.96 (d, J=1.9 Hz, 1H), 7.47 (d, J= 15.4 Hz, 1H), 7.37 (d, J=2.0 Hz, 1H), 7.31 (d, J=7.8 Hz, 1H), 7.13 (t, J=7, 9 Hz, 2H), 6.86 (d, J=8.4 Hz, 1H), 6.79 (dd, J=8.0 Hz, 0.9 Hz, 1H), 6.70 (d, J =2.4 Hz, 1H), 6.51 (dd, J=8.4 Hz, 2.5 Hz, 1H), 6.06-6.02 (m, 1H), 6.02 (s, 2H ), 4.92 (d, J=1.1 Hz, 1H), 4.78 (s, 2H), 3.25-3.13 (m, 2H), 3.00 (d, J=1, 1 Hz, 3H), 2.49 (s, 3H), 1.29 (s, 3H).

Example 37. Synthesis of (E)-M-((7-(indolin-5-yloxy)-2-methylbenzofuran-3-yl)methyl)-M-methyl-3-(8oxo-6,7,8,9- tetrahydro-5H-pyrido[2,3-b]azepin-3-yl)acrylamide (compound 180).

General synthesis scheme.

Reaction conditions: a) CuI, N,N-dimethylglycine, Cs ₂ CO ₃ , DMF, 100°C; b) Pd/C, MeOH, _H2 , 50°C; c) GATU, DIPEA, DMF; d) TPA, DXM.

Step 1. tert-Butyl 5-((3-(((4-methoxybenzyl)(methyl)amino)methyl)-2-methylbenzofuran-7yl)oxy)indoline-1-carboxylate (compound 177).

Mixture of 3-(((4-methoxybenzyl)(methyl)amino)methyl)-2-methylbenzofuran-7-ol 76 (150 mg,

0.48 mmol), copper(1) iodide (92 mg, 0.48 mmol), N,N-dimethylglycine (50 mg, 0.48 mmol), Cs ₂ CO ₃ (628 mg, 1.9 mmol) and tert-butyl 5-bromoindoline-1-carboxylate 176 (144 mg, 0.48 mmol) was evacuated and backfilled with nitrogen (3x). DMF (3 ml) is added and the mixture is heated to 110°C overnight. The mixture is diluted with water (5 ml) and EtOAc (5 ml). The phases are separated and the aqueous phase is extracted with EtOAc (2x10 ml). The combined organic phases are washed with brine (15 ml), dried over MgSO ₄ , filtered and concentrated in vacuo. The crude product was purified by chromatography (0-100% EtOAc/isohexane) to give the title compound 177 as a dark yellow oil (129 mg, 49%). R _t 3.21 min (method 1b); m/z 529 [M+H] ⁺ (ER ⁺ ). 1H NMR (500 MHz, CDC13): δ, ppm 7.47-7.24 (m, 4H), 7.09 (t, J=7.8 Hz, 1H), 6.97-6.87 (m, 4H), 6.71 (d, J=7.9 Hz, 1H), 4.02 (shs, 2H), 3.83 (s, 3H), 3.56 (s, 2H), 3 .52 (s, 2H), 3.08 (t, J=8.6 Hz, 2H), 2.46 (s, 3H), 2.21 (s, 3H), 1.59 (s, 9H) .

177 178

Step 2. tert-Butyl 5-((2-methyl-3-((methylamino)methyl)benzofuran-7-yl)oxy)indoline-1carboxylate (compound 178).

A solution of tert-butyl 5-((3-(((4-methoxybenzyl)(methyl)amino)methyl)-2-methylbenzofuran-7yl)oxy)indoline-1-carboxylate 177 (129 mg, 0.24 mmol) in MeOH (5 ml) hydrogenate in H-Cube (10% Pd/C,

30x4 mm, full hydrogen, 50°C, 1 ml/min). The solvent was removed in vacuo, then the residue was redissolved in MeOH and loaded onto an SCC column (1 g). The column is washed with MeOH (50 ml) and then the product is eluted with 0.7 M methanolic ammonia and concentrated in vacuo. The crude product was purified by chromatography (0-10% (0.7 M NH ₃ /MeOH)/DCM) to give the title compound 178 as a colorless oil (54 mg, 52%). Rt 2.41 min (method 1b); m/z 322 (M - tBu-NHCH ₃ ) ⁺ (ER ⁺ ).

Stage 3. (E)-tert-Butyl

5-((2-methyl-3-((X-methyl-3-(8-oxo-6,7,8,9-tetrahydro-5H-pyrido[2,3b]azepin-3-yl)acrylamido)methyl )benzofuran-7-yl)oxy)indoline-1-carboxylate (compound 179).

To a suspension of tert-butyl 5-((2-methyl-3-((methylamino)methyl)benzofuran-7-yl)oxy)indoline-1carboxylate 178 (100 mg, 0.24 mmol), trifluoroacetate (E)-3- (8-oxo-6,7,8,9-tetrahydro-5H-pyrido[2,3b]azepin-3-yl)acrylic acid 4 (109 mg, 0.24 mmol) and DIPEA (0.21 ml, 1 .22 mmol) in DMF (2.0 ml) was added GATU (102 mg, 0.27 mmol) and the reaction mixture was stirred for 2 hours. Then water (3.0 ml) was added and the solid was collected by filtration. The crude solid was purified by chromatography (0-10% MeOH/DCM) to give the title compound 179 as a yellow solid (100 mg, 62%). Rt 2.63 min (method 1a); m/z 623 [M+H] ⁺ (ER ⁺ ).

Step 4. (E)-N-((7-(Indolin-5-yloxy)-2-methylbenzofuran-3-yl)methyl)-N-methyl-3-(8-oxo-6,7,8,9tetrahydro -5H-pyrido[2,3-b]azepin-3-yl)acrylamide (compound 180).

TFA (2 ml, 25 mmol) is added dropwise to a stirred solution of N-tert-butyl 5-((2methyl-3-((N-methyl-3-(8-oxo-6,7,8,9-tetrahydro- 5H-pyrido[2,3-b]azepin-3-yl)acrylamido)methyl)benzofuran-7-yl)oxy)indoline-1-carboxylate 179 (100 mg, 0.16 mmol) in DCM (4.0 ml ). The reaction mixture is stirred for 30 minutes and then concentrated in vacuo. The resulting yellow oil was suspended in NaHCO ₃ (10 ml) and stirred for 30 minutes. The aqueous mixture was extracted with DCM (3x10 ml) and the combined organic layers were dried with MgSO4, filtered and concentrated in vacuo to give the title compound 180 as a beige solid (50 mg, 56%). Rt 1.36 min (method 1a); m/z 523 [M+H] ⁺ (ER ⁺ ). ^1H NMR (500 MHz, DMSO-06): δ, ppm 9.57 (s, 1H), 8.49 (d, J=2.2 Hz, 1H), 8.03 (d, J= 2.2 Hz, 1H), 7.57 (d, J=15.4 Hz, 1H), 7.36-7.18 (m, 2H), 7.07 (t, J=7.9 Hz, 1H), 6.87-6.79 (m, 1H), 6.67 (dd, J=8.4 Hz, 2.5 Hz, 2H), 6.50 (d, J=8.3 Hz, 1H), 5.12 (s, 1H), 4.79 (s, 2H), 3.46 (t, J=8.5 Hz, 2H), 3.01 (s, 3H), 2.93 ( t, J=8.5 Hz, 2H), 2.77 (t, J=7.2 Hz, 2H), 2.50 (s, 3H), 2.32 (t, J=7.2 Hz, 2H), 2.15 (p, J=7.2 Hz, 2H).

Example 38. Synthesis of (K,E-N-((7-((2,3-dihydrobenzofuran-6-yl)oxy)-2-methylbenzofuran-3yl)methyl)-3-(3-hydroxy-3-methyl- 4-oxo-2,3,4,5-tetrahydro-1H-pyrido[2,3-b][1,4]diazepin-8-yl)-N-methylacrylamide (compound 182).

General synthesis scheme.

Reagents and conditions: a) acryloyl chloride, TEA, 0°C to RT; b) Pd-162, MeNCy ₂ , NBu ₄ Cl, 1,4-dioxane, 80°C.

Step 1. N((7-((2,3-Dihydrobenzofuran-6-yl)oxy)-2-methylbenzofuran-3-yl)methyl)-N-methylacrylamide (compound 181).

To an ice-cold solution of 1-(7-((2,3-dihydrobenzofuran-6-yl)oxy)-2-methylbenzofuran-3-yl)-Nmethylmethanamine 162 (160 mg, 0.52 mmol) and TEA (300 μl, 2.07 mmol) in DCM (10 ml) added dropwise

- 103 046887 lyam solution of acryloyl chloride (42 µl, 0.52 mmol) in DCM (1.0 ml). The reaction mixture is warmed to RT and stirred for 1 hour. The reaction mixture is quenched with water (5.0 ml) and the two phases are separated. The organic phase is dried with MgSO ₄ , filtered and concentrated in vacuo. The crude product was purified by chromatography (0-100% EtOAc/isohexane) to give the title compound 181 as a colorless oil (120 mg, 60%). Rt 2.36 min (method 1a); m/z 364 [M+H]+ (ER ⁺ ).

Stage 2. ^E-N-((7-((2,3-Dihydrobenzofuran-6-yl)oxy)-2-methylbenzofuran-3-yl)methyl)-3-(3hydroxy-3-methyl-4-oxo -2,3,4,5-tetrahydro-1H-pyrido[2,3-b][1,4]diazepin-8-yl)<N-methylacrylamide (compound 182).

Na((7-((2,3-dihydrobenzofuran-6-yl)oxy)-2methylbenzofuran-3-yl)methyl)-N-methylacrylamide 181 (60 mg, 0.17 mmol), A)- is loaded into the reaction tube 8-bromo-3-hydroxy-3methyl-2,3-dihydro-1H-pyrido[2,3-b][1,4]diazepin-4(5H)-one 47 (45 mg, 0.17 mmol), NBu ₄ Cl (5 mg, 0.02 mmol), [P(tBu) ₃ ]Pd(crotyl)Cl (Pd-162) (7 mg, 0.02 mmol) and the tube were washed with N2 for 5 min. 1,4-Dioxane (3.0 mL) and N-cyclohexyl-N-methylcyclohexanamine (75 μL, 0.35 mmol) were added and the reaction mixture was purged with N2 for a further 5 min. The mixture is heated to 80°C for 1 hour and then cooled to RT. Ethyl acetate (5.0 ml) was added and the resulting solution was washed with water (5.0 ml) and brine (5.0 ml), dried over MgSO ₄ , filtered and concentrated in vacuo. The crude product was purified by chromatography (0-10% MeOH/DCM) to give the title compound 182 as a pale yellow solid (29 mg, 36%). Rt 2.12 min (method 1a); m/z 555 [M+H] ⁺ (ER ⁺ ). ^1H NMR (500 MHz, DMSO-ph, 363 K): δ, ppm 9.38 (s, 1H), 7.96 (d, J=1.9 Hz, 1H), 7.47 (d , J=15.4 Hz, 1H), 7.41-7.25 (m, 2H), 7.25-7.03 (m, 3H), 6.83 (dd, J=7.9 Hz, 1.1 Hz, 1H), 6.47 (dd, J=8.0 Hz, 1.6 Hz, 1H), 6.42 (d, J=2.2 Hz, 1H), 6.05 (s , 1H), 4.93 (s, 1H), 4.78 (s, 2H), 4.57 (t, J=8.6 Hz, 2H), 3.25-3.11 (m, 4H) , 3.03 (s, 3H), 2.48 (s, 3H), 1.29 (s, 3H).

Example 39. Synthesis of N,E-3-(3-hydroxy-3-methyl-4-oxo-2,3,4,5-tetrahydro-III-pyrido[2,3b][1,4]diazepine-8- yl)-N-((7-(3-methoxyphenoxy)-2-methylbenzofuran-3-yl)methyl)-N-methylacrylamide (compound 183).

General synthesis scheme.

153 142

183

Reagents and conditions: a) HOBt-H ₂ O, EDC-HCl, DIPEA, DMF.

Step 1. (R,E-3 -(3-Hydroxy-3-methyl-4-oxo-2,3,4,5-tetrahydro-1H-pyrido[2,3-b][1,4]diazepine -8yl)-N-((7-(3-methoxyphenoxy)-2-methylbenzofuran-3-yl)methyl)-N-methylacrylamide (compound 183).

A microwave tube covered with aluminum foil is loaded with HOBt- _H2O (8.04 mg, 0.05 mmol) and (E)-3-[(3R)-3-hydroxy-3-methyl-4-oxo-2. 5-dihydro-1H-pyrido[2,3-b][1,4]diazepin-8yl]prop-2-enoic acid 142 (9.21 mg, 0.04 mmol) and DMF (0.3 ml) . Add 1-[7-(3methoxyphenoxy)-2-methyl-benzofuran-3-yl]-N-methyl-methanamine 153 (10.41 mg, 0.04 mmol) and EDCl-HCl (10.06 mg, 0. 05 mmol), and after 2 min of stirring at RT, DIPEA (0.02 ml, 0.12 mmol) was added. The reaction mixture is stirred in the dark overnight at RT. Then water and acetonitrile are added, the mixture is treated with ultrasound and the precipitate is filtered off. The crude product (24 mg) was purified by preparative HPLC using a 100% to 100% water/ACN gradient to give the title compound 183 as a yellow oil (3.7 mg, 19.4% yield). Rt 1.83 min (method 1b); m/z 543.5 [M+H] ⁺ (ER ⁺ ). 1H NMR (400 MHz, DMSO-ph): δ, ppm 9.93 (s, 1H), 7.98 (s, 1H), 7.52-7.17 (m, 5H), 7.06 (d, J=15.5 Hz, 1H), 6.88 (d, J=8.2 Hz, 1H), 6.71 (dd, J=1.7 Hz, 7.9 Hz, 1H), 6.61-6.56 (m, 1H), 6.51 (dd, J=1.8 Hz, 8.4 Hz, 1H), 6.28-6.19 (m, 1H), 5.28 (s, 1H), 4.90, 4.75 (2 s, 2H), 3.73 (s, 3H), 3.21-3.08 (m, 2H), 3.06, 2.96 ( 2 s, 3H), 2.45 (s, 3H), 1.23 (s, 3H) (rotamers).

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Example 40. Synthesis of (E)-Na-((7-(3-methoxyphenoxy)-2-methylbenzofuran-3-yl)methyl)-Na-methyl-3-(8oxo-6,7,8,9-tetrahydro- 5H-pyrido[2,3-b]azepin-3-yl)acrylamide (compound 186).

General synthesis scheme.

Reagents and conditions: a) acryloyl chloride, TEA, DCM, 0°C; b) Pd-116, DIPEA, NBu ₄ Cl, dioxane, 80°C.

The synthesis of 3-bromo-6,7-dihydro-5H-pyrido[2,3-b]azepin-8(9H)-one (compound 185) was previously described in AFFINIUM PHARMECEUTICALS, INC: WO2007/67416, 2007, A2.

Step 1. M((7-(3-Methoxyphenoxy)-2-methylbenzofuran-3-yl)methyl)-Mmethylacrylamide (compound 184).

To a stirred solution of 1-(7-(3-methoxyphenoxy)-2-methylbenzofuran-3-yl)-Nmethylmethanamine 153 (155 mg, 0.52 mmol) in DCM (1.5 ml) add TEA (0.22 ml , 1.58 mmol) and acryloyl chloride (0.051 ml, 0.63 mmol) at 0°C. The reaction mixture is stirred at 0°C for 1 hour, then heated at RT. The reaction mixture was quenched with water (5.0 ml), then extracted into DCM (20 ml). The organics are separated and dried by passing through a hydrophobic frit, and then concentrated in vacuo. The crude product was purified by silica chromatography (0-100% EtOAc/isohexane) to give the title compound 184 as a yellow oil (122 mg, 67%). Rt 2.32 min (method 1b); m/z 352 [M+H]+ (ER ⁺ ).

\Y = Pd-116, DIPEA

0-YjG YQ ----/— ^y ° XN pjYs Bu ₄ nci, dioxane, so c

_MeOHq

184 185

Step 2. (E)-N-((7-(3-Methoxyphenoxy)-2-methylbenzofuran-3-yl)methyl)-N-methyl-3-(8-oxo-6,7,8,9tetrahydro-5H -pyrido[2,3-b]azepin-3-yl)acrylamide (compound 186).

N((7-(3-methoxyphenoxy)-2-methylbenzofuran-3-yl)methyl)-Nmethylacrylamide 184 (55 mg, 0.16 mmol), 3-bromo-6,7-dihydro-5H is loaded into the reaction tube -pyrido[2,3-b]azepin-8(9H)-one 185 (38 mg, 0.16 mmol), n-Bu ₄ NCl-H ₂ O (5.0 mg, 0.02 mmol) and Pd [P(tBu) ₃ ] ₂ (Pd-116) (8.00 mg, 0.02 mmol) and the tube is evacuated and backfilled with N2 (3x). 1,4-Dioxane (4.0 mL) and DIPEA (55 μL, 0.30 mmol) were added and the reaction mixture was heated to 80°C for 30 min. The reaction mixture was evaporated to dryness and the crude product was purified by chromatography (0-10% MeOH/DCM) to give the title compound 186 as a pale yellow solid (42 mg, 52%). R _t 2.26 min (method 1a); m/z 512 [M+H] ⁺ (ER ⁺ ). ^1H NMR (500 MHz, DMSO-06, 363 K): δ, ppm 9.57 (s, 1H), 8.49 (d, J=2.2 Hz, 1H), 8.04 (d , J=2.2 Hz, 1H), 7.57 (d, J=15.5 Hz, 1H), 7.37 (d, J=7.8 Hz, 1H), 7.26 (t, J =8.2 Hz, 2H), 7.17 (t, J=7.9 Hz, 1H), 6.88 (d, J=7.9 Hz, 1H), 6.72 (dd, J=8 .3 Hz, 2.3 Hz, 1H), 6.60 (d, J=2.5 Hz, 1H), 6.56 (dd, J=8.3 Hz, 2.2 Hz, 1H), 4 .80 (s, 2H), 3.76 (s, 3H), 3.00 (s, 3H), 2.77 (t, J=7.2 Hz, 2H), 2.48 (s, 3H) , 2.32 (t, J=7.2 Hz, 2H), 2.16 (t, J=7.2 Hz, 2H).

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Example 41. Synthesis of (E)-N-((7-(3-methoxyphenoxy)-2-methylbenzo[b]thiophen-3-yl)methyl)-N-methyl-3(8-oxo-6,7,8 ,9-tetrahydro-5H-pyrido[2,3-b]azepin-3-yl)acrylamide (compound 194).

General synthesis scheme.

Reaction conditions: a) dichloro(methoxy)methane, tin chloride (HC), DCM, 0°C; b) 1-(4-methoxyphenyl)-Nmetalmethanamine, Na(OAc)3BH, DCE; c) Pd-175, tBuBrettPhosm KOH, 1,4-dioxane, H2O, 80°C; d) 3bromoanisole 191, CuI, N^dimethylglycine, CS2CO3, DMF, 110°C; e) ACE-Ci, MeOH, 0°C to RT; f) GATU, DIPEA, DMF.

Step 1. 7-Bromo-2-methylbenzo[b]thiophene-3-carbaldehyde (compound 188). To a solution of 7-bromo-2methylbenzothiophene 187 (950 mg, 4.18 mmol) and dichloro(methoxy)methane (0.42 ml, 4.64 mmol) in DCM (20 ml) at 0°C add a solution dropwise tin chloride (GU) (1 M in DCM, 5 ml, 5.00 mmol) for 15 min. After addition is complete, the mixture is heated to RT for 1 hour, then poured into ice-cold brine. NaHCO ₃ solution (50 ml). The mixture is extracted into DCM (2x100 ml), the organic extracts are combined, dried over Na ₂ SO ₄ , filtered and concentrated in vacuo. The crude product was purified by chromatography (0-10% EtOAc/isohexane) to give the title compound 188 as a white solid (780 mg, 69%). ¹ H NMR (500 MHz, CDCl ₃ ): δ, ppm 10.37 (s, 1H), 8.58 (dd, J=8.2 Hz, 1.1 Hz, 1H), 7.55 ( dd, J=7.6 Hz, 1.1 Hz, 1H), 7.39-7.35 (m, 1H), 2.97 (s, 3H).

Step 2. 1-(7-Bromo-2-methylbenzo[b]thiophen-3-yl)-N-(4-methoxybenzyl)-N-methylmethanamine (compound 189).

Sodium triacetoxyborohydride (1.61 g, 7.74 mmol) was added in one portion to a stirred solution of 7-bromo-2-methylbenzo[b]thiophene-3-carbaldehyde 188 (790 mg, 3.10 mmol) and 1-(4- methoxyphenyl)N-methylmethanamine 70 (0.46 ml, 3.07 mmol) in EDC (12 ml) at RT. The reaction mixture was stirred for ~16 h, then the solvent was concentrated in vacuo and the resulting residue was taken up in NaHCO ₃ (25 ml, sat. aq.). The aqueous product was extracted with DCM (3x30 ml) and the combined organic layers were washed with brine (30 ml), then dried through a phase separator cartridge. The crude product was purified by chromatography (0-50% EtOAc/isohexane) to give the title compound 189 as a brown oil (1.05 g, 86%). R _t 3.18 min (method 1b); m/z 390/392 [M+H]+ (ER+).

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Step 3. 3-(((4-Methoxybenzyl)(methyl)amino)methyl)-2-methylbenzo[b]thiophen-7-ol (compound 190).

1-(7-bromo-2-methylbenzo[b]thiophen-3-yl)-H-(4-methoxybenzyl)-H-methylmethanamine 189 (1.05 g, 2.69 mmol), Pd-175 is charged into the flask (0.04 g, 0.05 mmol), tBuBrettPhos (0.03 g, 0.05 mmol) and KOH (0.45 g, 8.08 mmol). It is evacuated/backfilled with nitrogen (3 times) and then 1,4-dioxane (5.0 ml) is added and degassed with water (0.8 ml). The resulting mixture is heated to 80°C and stirred for 3 hours. The mixture is cooled to RT and diluted with EtOAc (20 ml), then acidified with 1 M HCl (20 ml). The mixture is stirred for 5 minutes until all the solid has dissolved. The mixture is treated with sat. aq. NaHCO ₃ (50 ml) and the phases are separated. The aqueous phase is extracted with EtOAc (2x50 ml). The combined organic extracts are dried over MgSO ₄ , filtered and concentrated. The crude product was purified by chromatography (0-50% EtOAc/isohexane) to give the title compound 190 as a brown oil (879 mg, 99%). R _t 2.51 min (method 1b); m/z 328 [M+H]+ (ER ⁺ ).

V. [<%G ^VG Cui, Ν,Ν-dimethylglycine, \,rmv

II * M -------- P- ^oXU \ 1 Cs ₂ CO ₃ , DMF, 110 °C ^and S-Ά.

o.o ⁴ \

190 ₁₉₁ 192

Step 4. N-(4-Methoxybenzyl)-1-(7-(3-methoxyphenoxy)-2-methylbenzo[Cthiophene-3-n.i)-Nmethylmethanamine (compound 192).

Mixture of 3-(((4-methoxybenzyl)(methyl)amino)methyl)-2-methylbenzo[b]thiophen-7-ol 190 (250 mg, 0.76 mmol), copper iodide) (150 mg, 0.76 mmol), 2-(dimethylamino)acetic acid (80 mg, 0.76 mmol) and Cs ₂ CO ₃ (1.0 g, 3.05 mmol) were evacuated and backfilled with nitrogen (3 times). DMF (5.0 ml) and 1-bromo-3-methoxybenzene 191 (0.12 ml, 0.92 mmol) were added and the mixture was heated to 110°C overnight. The mixture is filtered through a pad of Celite®, washed with EtOAc (40 ml). The filtrate is evaporated to dryness. The crude product was purified by chromatography (0-30% EtOAc/isohexane) to give the title compound 192 as a colorless oil (287 mg, 86%). R _t 3.15 min (method 1b); m/z 434 [M+H]+ (ER ⁺ ).

Step 5. 1-(7-(3-Methoxyphenoxy)-2-methylbenzo[b]thiophen-3-yl)-N-methylmethanamine (compound 193).

1-Chloroethyl chloroformate (0.15 mL, 1.37 mmol) was added dropwise to the stirred solution of N-(4-methoxybenzyl)-1-(7-(3-methoxyphenoxy)-2-methylbenzo[b]thiophen-3-yl )-N-methylmethanamine 192 (290 mg, 0.66 mmol) in DCM (9.0 ml) at 0°C under N ₂ . The reaction mixture is warmed to RT and stirred overnight. The solvent is then concentrated in vacuo and the residue is redissolved in MeOH (15 ml) and heated to reflux for 1 hour. The mixture is cooled to RT and applied to an SQL column. The column was washed with methanol (50 ml), the product was eluted with 10% methanolic ammonia and evaporated to give the title compound 193 as a dark oil (122 mg, 51%). R _t 2.11 min (method 1b); m/z 283 [M-NHCH ₃ ] ⁺ (ER ⁺ ).

193 4 1 94

Step 6. (E)-N-((7-(3-Methoxyphenoxy)-2-methylbenzo[b]thiophen-3-yl)methyl)-N-methyl-3-(8-oxo6,7,8,9 -tetrahydro-5H-pyrido[2,3-b]αzepin-3-yl)acrylamide (compound 194).

Suspension of 1-(7-(3-methoxyphenoxy)-2-methylbenzo[b]thiophen-3-yl)-N-methylmethanamine 193 (64 mg, 0.20 mmol), (E)-3-(8-oxo- 6,7,8,9-tetrahydro-5H-nirido[2,3-b]αzenin-3-yl)acrylic acid 4 (91 mg, 0.20 mmol), DIPEA (0.18 ml, 1.03 mmol ) in DMF (1.0 ml) and stir for 10 minutes. GATU (78 mg, 0.20 mmol) was added in one portion and the reaction mixture was stirred for 1 hour. The mixture was diluted with water (10 ml), extracted with DCM (3x20 ml), washed with brine, passed through a phase separator and evaporated. The crude product is purified by chromatography (0-3% MeOH/DCM). Fractions were collected then evaporated and the solid triturated with MeCN to give the title compound 194 as a white solid (21 mg, 19%). Rt 2.36 min (method 1b); m/z 528 [M+H]+ (ER+). ^1H NMR (500 MHz, DMSO<): δ, ppm 9.57 (s, 1H), 8.49 (d, J=2.1 Hz, 1H), 8.03 (d, J=2 .2 Hz, 1H), 7.62 (d, J=8.0 Hz, 1H), 7.58 (d, J=15.4 Hz, 1H), 7.35 (t, J=7.9 Hz, 1H), 7.30-7.27 (m, 2H), 6.90 (d, J=7.8 Hz, 1H), 6.76 (dd, J=8.3 Hz, 2.3 Hz, 1H), 6.64-6.63 (m, 1H), 6.60 (dd, J=8.2 Hz, 2.3 Hz, 1H), 4.94 (s, 2H), 3, 76 (s, 3H), 2.96 (s, 3H), 2.77 (t, J=7.2 Hz, 2H), 2.59 (s, 3H), 2.32 (t, J=7 .2 Hz, 2H), 2.18-2.12 (m, 2H).

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Example 42 Synthesis of 4-(4-((3aR,6aS)-2-((E)-3-(4-OKCO-2,3,4,5-TeTparugpo-1H-nupugo[2,3b][1, 4]diazepin-8-yl)acryloyl)-1,2,3,3a,4,6a-hexahydrocyclopenta[c]pyrrol-5yl)phenoxy)benzonitrile (compound 198).

General synthesis scheme.

198

Reaction conditions: a) K2CO3, H2O, PdCl ₂ (dppf), 1,4-dioxane, 80°C; b) TPA, DXM; c) GATU, DIPEA, DMF.

The synthesis of 4-(4-(4,4,5-trimethyl-1,3,2-dioxaborolan-2-yl)phenoxy)benzonitrile (compound 195) was previously described by Abbott Laboratories; US2002/156081, 2002, A1 and X-RX DISCOVERY, INC.; WO2015/48662, 2015, A2. Pinacol ester 195 was prepared in 90% yield with a purity of 93% (R _t =2.40 min (standard); m/z 321.1 [M+H]+ (ER+)).

Steps 1 and 2. 4-(4-((3aS,6aR)-1,2,3,3a,4,6a-hexahydrocyclopenta[c]pyrrol-5yl)phenoxy)benzonitrile 2,2,2-Trifluoroacetate (compound 197 ) is synthesized from compounds 57 and 195 via intermediate 196 in 2 steps according to the procedure described in example 20 and US2017/0174683 A1. The crude product (TPA salt) 197 is used in the next step without purification (R _t =1.26 min (standard); m/z 303.6 [M+H]+ (ER+)).

Step 3. 4-(4-((3aR,6aS)-2-((E)-3-(4-Oxo-2,3,4,5-tetrahydro-1H-pyrido[2,3-b][ 1,4]diazepin-8yl)acryloyl)-1,2,3,3a,4,6a-hexahydrocyclopenta[c]pyrrol-5-yl)phenoxy)benzonitrile (compound 198) is prepared from acid 6 and amine 197 as yellow solid with a purity of 99% (19% yield) by the standard amine-amide coupling reaction described in example 20. R _t =1.78 min (standard); m/z 518.3 [M+H]+ (ER+)). ¹ H NMR (400 MHz, DMSO-dg): δ, ppm 9.73 (d, J=4.9 Hz, 1H), 7.98 (d, J=6.2 Hz, 1H), 7 .84 (d, J=8.8 Hz, 2H), 7.58 (d, J=8.3 Hz, 2H), 7.39-7.28 (m, 2H), 7.16-7, 05 (4H, m), 6.86 (dd, J=6.7 Hz, 15.5 Hz, 1H), 6.20 (s, 1H), 6.05-6.00 (m, 1H), 4.05-3.75 (m, 2H), 3.73-3.49 (m, 2H), 3.47-3.36 (m, 3H), 3.22-3.10 (m, 1H ), 3.10-2.91 (m, 1H), 2.69-2.55 (m, 3H).

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Example 43. Synthesis of (E)-3-(3-hydroxy-2,3-dimethyl-4-oxo-2,3,4,5-tetrahydro-1H-pyrido[2,3b][1,4]diazepine- 8-yl)-Nmethyl-N((3-methylbenzofuran-2-yl)methyl)acrylamide (compound 207).

General synthesis scheme.

199 200 201 202 mixture of diastereomers mixture of diastereomers 3:2 diastereomer ratio

first eluted diastereomer 203

second eluted diathoreomer 204

Reaction conditions: a) nitroethane, Amberlyst A-21, 0°C to RT; b) Zn, acetic acid; c) 5-bromo-3fluoro-2-nitropyridine 11, TEA, EtOH, reflux; d) Fe, NH ₄ Cl, EtOH, H ₂ O, reflux, separation of diastereomers; e) LiOH, H ₂ O, THF, MeOH; f) GATU, DIPEA, DMF; g) Pd-116, DIPEA, NBu ₄ Cl, 1,4-dioxane, 80°C. nitroethane, θΕ Amberlyst A-21 0 °C to KT

199 200 mixture of diastereomers

Step 1. Ethyl 2-hydroxy-2-methyl-3-nitrobutanoate (compound 200).

Ethyl 2-oxopropanoate 199 (15.6 mL, 140 mmol) was added dropwise to a stirred suspension of Amberlyst A-21 (14 g, 140 mmol) in nitroethane (10.0 mL, 140 mmol) at 0°C. The reaction mixture is warmed to RT and stirred for ~16 hours. The resin is removed by filtration and washed with DCM (3x100 ml). The filtrate was concentrated in vacuo to give the desired product 200 as a yellow oil (20.6 g, 73%) in an approximately 2:1 diastereomer ratio.

Main diastereomer: ^1H NMR (500 MHz, DMSO-d^: δ, ppm 5.94 (s, 1H), 5.04 (kv, J=6.8 Hz, 1H), 4.17 (kv , J=7.1 Hz, 2H), 1.46 (d, J=6.8 Hz, 3H), 1.31 (s, 3H), 1.22 (t, J=7.1 Hz, 3H ).

Minor diastereomer: 1H NMR (500 MHz, DMSO^ ₆ ): δ, ppm 6.08 (s, 1H), 4.88 (sq, J=6.8 Hz, 1H), 4.17 (sq, J=7.1 Hz, 2H), 1.47 (d, J=6.8 Hz, 3H), 1.36 (s, 3H), 1.22 (t, J=7.1 Hz, 3H) .

HO / Zn, acetic acid HO /

IO,N_X.OEt H ₂ N_X_.OEt 'o I O

200 201 mixture of diastereomers mixture of diastereomers

Step 2. Ethyl 3-amino-2-hydroxy-2-methylbutanoate (compound 201).

To a solution of ethyl 2-hydroxy-2-methyl-3-nitrobutanoate 200 (5 g, 26.2 mmol) in acetic acid (300 ml), add zinc powder in portions (68.4 g, 1046 mmol). The mixture is stirred at RT for 4 hours, then filtered over a layer of Celite®. The filtrate is neutralized by adding sat. aq. Na ₂ CO ₃ (~700 ml) and extract with EtOAc (2x700 ml). The combined organic layers are dried over Na ₂ SO ₄ , filtered and evaporated under reduced pressure to give the title compound 201c

- 109 046887 in the form of yellow oil. The aqueous layer is then evaporated and the residue is divided into two portions, each of which is solubilized again in water (300 ml). The aqueous product is filtered to remove the formed salts, extracted with EtOAc (2x400 ml) and DCM (2x400 ml), and the combined organic layers are dried over MgSO ₄ , filtered and evaporated to dryness. 1H NMR analysis of the three fractions matched the structure of the product to 100% purity and were combined to give the title compound 201 as a yellow oil (1.12 g, 27%). 1H NMR (DMSO-dg): δ, ppm δ 4.12-4.07 (m, 2H), 2.94-2.84 (m, 1H), 1.24-1.15 (m, 6H), 0.94-0.91 (m, 3H). NH2 and OH are not observed (mixture of isomers).

mixture of isomers 2:3 diastereomer ratio

Step 3. Ethyl 3-((5-bromo-2-nitropyridin-3-yl)amino)-2-hydroxy-2-methylbutanoate (compound 202).

Triethylamine (1.8 ml, 12.8 mmol) was added to a stirred solution of 5-bromo-3-fluoro-2nitropyridine 11 (1.3 g, 5.88 mmol) and ethyl 3-amino-2-hydroxy-2-methylbutanoate 201 (1.06 g, 4.27 mmol) in EtOH (20 mL), and the reaction mixture was heated to reflux for ~16 h. The reaction mixture was cooled to RT, then concentrated in vacuo, and the crude product was purified by column chromatography (0-10% MeOH/DCM) to give the desired product 202 as a yellow oil (1.24 g, 79%) with an approximately 3:2 diastereomer ratio.

Diastereomer 1: Rt 1.90 min (method 1a) m/z 362/364 [M+H]+ (ER+). 1H NMR (400 MHz, DMSO- ₀₆ ): δ, ppm 8.11-8.02 (m, 2H), 7.91-7.84 (m, 1H), 5.96 (s, 1H ), 4.31-4.23 (m, 1H), 4.00 (kv, J=7.1 Hz, 2H), 1.34 (s, 3H), 1.17 (d, J=6, 4 Hz, 3H), 1.01 (t, J=7.1 Hz, 3H).

Diastereomer 2: R _t 1.96 min (method 1a) m/z 362/364 [M+H]+ (ER ⁺ ). 1H NMR (400 MHz, DMSO-dg) δ 8.13-8.00 (m, 2H), 7.89-7.85 (m, 1H), 5.98 (s, 1H), 4.18 ( kv, J=7.1 Hz, 2H), 4.15-4.08 (m, 1H), 1.34 (s, 3H), 1.25 (t, J=7.1 Hz, 3H), 1.09 (d, J=6.4 Hz, 3H).

202 203 204

2:3 ratio of diastereomers first eluted second eluted diastereomer diastereomer

Step 4. Ethyl 3-((2-amino-5-bromopyridin-3-yl)amino)-2-hydroxy-2-methylbutanoate (compounds 203 and 204).

Mixture of ethyl 3-((5-bromo-2-nitropyridin-3-yl)amino)-2-hydroxy-2-methylbutanoate 202 (1.2 g, 3.31 mmol), iron (1.48 g, 26, 5 mmol) and NH ₄ Cl (0.71 g, 13.3 mmol) in a mixture of EtOH (20 ml) and H2O (5 ml) are heated and stirred at reflux for ~16 hours. The reaction mixture is cooled to CT, then loaded dry onto Celite® and purified by column chromatography (0-50% EtOAc/isohexane) to give the desired product as a brown solid: Diastereomer 1,203 (154 mg, 14%) and Diastereomer 2,204 (280 mg, 25 %).

Diastereomer 1 203: Rt 1.14 min (method 1a) m/z 332/334 [M+H] ⁺ (ER ⁺ ). 1H NMR (400 MHz, DMSO d ₆ ): δ, ppm 7.28 (d, J=2.1 Hz, 1H), 6.80 (d, J=2.1 Hz, 1H), 5, 63 (s, 2H), 5.11 (s, 1H), 4.39 (d, J=10.1 Hz, 1H), 3.96 (kv, J=7.0 Hz, 2H), 3, 86-3.78 (m, 1H), 1.28 (s, 3H), 1.07 (d, J=6.5 Hz, 3H), 1.03 (t, J=7.1 Hz, 3H ).

Diastereomer 2 204: Rt 1.13 min (method 1a) m/z 332/334 [M+h] ⁺ (ER ⁺ ). 1H NMR (400 MHz, DMSO d ₆ ): δ, ppm 7.27 (d, J=2.0 Hz, 1H), 6.71 (d, J=2.0 Hz, 1H), 5, 77 (s, 2H), 5.44 (s, 1H), 4.48 (d, J=9.7 Hz, 1H),

4.05 (kv, J=7.1 Hz, 2H), 3.76-3.67 (m, 1H), 1.29 (s, 3H), 1.16 (t, J=7.1 Hz , 3H), 1.06 (d, J=6.6 Hz, 3H).

Step 5. 3-((2-Amino-5-bromopyridin-3-yl)amino)-2-hydroxy-2-methylbutanoic acid hydrochloride (compound 205).

Lithium hydroxide (1 M aq, 0.6 mL, 0.6 mmol) was added dropwise to a stirred solution of ethyl 3-((2-amino-5-bromopyridin-3-yl)amino)-2-hydroxy-2- methyl butanoate 203 (65 mg, 0.20 mmol) in THF (5 ml) and MeOH (1 ml) at RT, and the reaction mixture was stirred for 4 hours. The mixture was acidified to ~pH 5 by adding 1 M HCl (aq) and the reaction mixture is concentrated in vacuo. The residue was azeotroped with MeCN (5 mL) to obtain the desired product 205 as a brown solid (68 mg, quant.). Rt 0.84 min (method 1a) m/z 304/306 [M+H] ⁺ (ER ⁺ ).

- 110 046887

Step 6. 8-Bromo-3-hydroxy-2,3-dimethyl-2,3-dihydro-1H-pyrido[2,3-b][1,4]diazepin-4(5H)-one (compound 206) .

DIPEA (0.13 mL, 0.76 mmol) was added to a stirred solution of 3-((2-amino5-bromopyridin-3-yl)amino)-2-hydroxy-2-methylbutanoic acid hydrochloride 205 (65 mg, 0.19 mmol) in DMF (2 ml) at RT. After 10 min, GATU (75 mg, 0.20 mmol) was added and the reaction mixture was stirred for ~16 h. The reaction mixture was quenched with MeOH (5 ml) and H2O (30 ml). The aqueous mixture was extracted with EtOAc (3x50 ml), and the combined organic layers were washed with brine (2x50 ml), dried with MgSO ₄ , and concentrated in vacuo. The crude product was purified by column chromatography (0-50% EtOAc/isohexane) to give the desired product 206 as a white solid (21 mg, 38%). Rt 1.21 min (method 2a) m/z 286/288 [M+H]+ (ER+). 1H NMR (DMSO-dy δ, ppm 10.25 (s, 1H), 7.66 (d, J=2.1 Hz, 1H), 7.26 (d, J=2.1 Hz, 1H ), 6.79 (d, J=6.4 Hz, 1H), 5.12 (s, 1H), 3.31-3.24 (m, 1H), 1.16 (s, 3H), 1 .03 (d, J=6.7 Hz, 3H). The cis-Stereochemistry of the resulting diastereomer is determined by NMR studies including NOE and ROESY experiments.

206 ²⁰⁷ racemic cis regioisomer

Step 7. (E)-3-(3-Hydroxy-2,3-dimethyl-4-oxo-2,3,4,5-tetrahydro-1H-pyrido[2,3-b][1,4]diazepine8 -yl)-U-methyl-U-((3-methylbenzofuran-2-yl)methyl)acrylamide (compound 207).

8-bromo-3-hydroxy-2,3-dimethyl-2,3-dihydro-1H-pyrido[2,3-b][1,4]diazepin4(5H)-one 206 (40 mg, 0 .14 mmol), Nmethyl-U-((3-methylbenzofuran-2-yl)methyl)acrylamide 9 (32 mg, 0.140 mmol), NBu ₄ Cl (4 mg, 0.01 mmol) and Pd-116 (7 mg, 0.01 mmol) and the flask was evacuated and backfilled with N2 three times. 1,4-Dioxane (2.5 ml) and DIPEA (0.05 ml, 0.28 mmol) were added and the reaction mixture was heated to 80°C and stirred for 3 hours. The solvent was removed in vacuo and the crude product was purified column chromatography (0-5% MeOH/DCM) to obtain the desired product 207 as a yellow solid (36 mg, 59%). Rt 1.93 min (method 1a) m/z 435 [M+H]+ (ER+). 1H NMR (DMSO-dfo 363 K): δ, ppm 9.72 (s, 1H), 7.95 (d, J=1.9 Hz, 1H), 7.59-7.53 (m, 1H), 7.49-7.35 (m, 3H), 7.32-7.21 (m, 2H), 7.12 (d, J=15.4 Hz, 1H), 6.32 (d , J=6.4 Hz, 1H), 4.84 (s, 2H), 4.79 (d, J=1.1 Hz, 1H), 3.39-3.30 (m, 1H), 3 .09 (s, 3H), 2.27 (s, 3H), 1.24 (s, 3H), 1.08 (d, J=6.7 Hz, 3H).

Example 44. Synthesis of N,E)-3-(2-(hydroxymethyl)-2-methyl-3-oxo-1,2,3,4-tetrahydropyrido[2,3b]pyrazin-7-yl)-N-methyl -N-((3-methylbenzofuran-2-yl)methyl)acrylamide (compound 208) and (R,E-3-(2(hydroxymethyl)-2-methyl-3-oxo-1,2,3,4- tetrahydropyrido[2,3-b]pyrazin-7-yl)-N-methyl-N-((3-methylbenzofuran-2-yl)methyl)acrylamide (compound 209).

Chiral resolution of compound 84

Chiral separation method: apparatus: Isolera (Biotage). Column: Chiralpak IA (20 µm; glass column; 250 mm x 25 mm). Eluent: acetonitrile/ethanol (8/2 v/v). Flow: 40 ml/min. Temperature: 25°C. Running time: 25 min.

- 111 046887

Injection quantity: 30 mg of compound 84 in 20 ml of eluent (dissolution at reflux, then cooling and rapid injection). Some product degradation is observed during hot dissolution, but no by-products are present in the fractions of interest.

^,E)-3-(2-(Hydroxymethyl)-2-methyl-3-oxo-1,2,3,4-tetrahydropyrido[2,3-b]pyrazin-7-yl)-N-methylN( (3-methylbenzofuran-2-yl)methyl)acrylamide (compound 208). Pure fractions of the first enantiomer are collected and concentrated until a precipitate is observed. Then evaporation to dryness is carried out under a stream of nitrogen at RT. The first enantiomer is obtained as a creamy powder (m=8.6 mg; chiral purity: 99.84%). The stereochemistry of the resulting enantiomer is determined arbitrarily.

(R,E-3-(2-(Hydroxymethyl)-2-methyl-3-oxo-1,2,3,4-tetrahydropyrido[2,3-b]pyrazin-7-yl)-N-methylN( (3-methylbenzofuran-2-yl)methyl)acrylamide (compound 209) Pure fractions of the second enantiomer are collected and concentrated until precipitation is observed. Evaporation to dryness is then carried out under a stream of nitrogen at RT. The first enantiomer is obtained as a creamy powder (m = 6). .9 mg; chiral purity: 99.59%). The stereochemistry of the resulting enantiomer is determined arbitrarily.

Example 45. Synthesis of N,E-Y-methyl-3-(2-methyl-4-oxo-2,3,4,5-tetrahydro-III-pyrido[2,3b][1,4]diazepine-8- yl)-K-((3-methylbenzofuran-2-yl)methyl)acrylamide (compound 214).

General synthesis scheme.

210 211 212 213

Reaction conditions: a) 5-bromo-3-fluoro-2-nitropyridine 11, TEA, EtOH, 80°C; b) Fe, NH4Cl, EtOH, water, 90°C; c) NaH, THF, 0°C to RT; d) Pd-116, NBu ₄ Cl, DIPEA, 1,4-dioxane, 80°C.

Step 1. ^)-Methyl 3-((5-bromo-2-nitropyridin-3-yl)amino)butanoate (compound 211).

To a stirred solution of 5-bromo-3-fluoro-2-nitropyridine 11 (430 mg, 1.95 mmol) in MeCN (10 ml) add N-methyl 3-aminobutanoate hydrochloride 210 (300 mg, 1.95 mmol), then TEA (700 µl, 4.88 mmol). The reaction mixture is stirred at reflux for 2 hours and then cooled to RT. The reaction mixture is concentrated in vacuo, then the crude product is dissolved in ethyl acetate (10 ml) and washed with water (10 ml). The organic layer is dried with MgSO ₄ , filtered and concentrated in vacuo. The crude product was purified by chromatography (0-100% EtOAc/isohexane) to give the title compound 211 as a yellow solid (590 mg, 90%). R _t 1.89 min (method 1a) m/z 318/320 [M+H]+ (ER+).

Vg

no ₂

211

OMe ^Fe NH ₄ CI, _Br

O EtOH:H ₂ O, 90 °C

.ОМе ¹ О NH ₂

212

Step 2. ^)-Methyl 3-((2-amino-5-bromopyridin-3-yl)amino)butanoate (compound 212).

To a mixture of methyl 3-((5-bromo-2-nitropyridin-3-yl)amino)butanoate 211 (570 mg, 1.85 mmol) in EtOH (35 ml) and water (8.75 ml) add iron (415 mg, 7.42 mmol) and ammonia hydrochloride (400 mg, 7.42 mmol). The resulting reaction mixture is heated and stirred at 90°C for 1 hour, then filtered through a pad of Celite® while remaining hot. The filtrate was evaporated to dryness and the crude product was purified by chromatography (0-10% MeOH/DCM) to give the title compound 212 as a colorless solid (430 mg, 76%). Rt 1.00 min (method 1a) m/z 288/290 [M+H]+ (ER+).

- 112 046887

Step 3. (K_)-8-Bromo-2-methyl-2,3-dihydro-'1I-pyrido[2,3-b]['1,4]diazepin-4(5I)-one (compound 213) .

Sodium hydride (60% in mineral oil, 83 mg, 2.08 mmol) is added in small portions to a stirred solution of methyl 3-((2-amino-5-bromopyridin-3-yl)amino)butanoate 212 (400 mg, 1. 39 mmol) in THF (20 ml) at 0°C. The reaction mixture is heated to RT and stirred for 2 hours. The reaction mixture is quenched by careful addition of sat. aq. NH4C1 (20 ml) and the aqueous mixture was extracted with EtOAc (3x25 ml). The combined organic layers are dried with MgSO ₄ and concentrated in vacuo. The crude product was purified by chromatography (0-10% MeOH/DCM) to give the title compound 213 as a colorless solid (170 mg, 45%). R _t 1.30 min (method 1a) m/z 256/258 [M+H]+ (ER+).

, o η - ¹ \ ?η π \ / I LΚΙ ·'

ΥΧΊ —---Ν ν4 Pd-116, Bu ₄ NCI, DIPEA, \_/Ν NV

Η O 1,4-dioxane, 90 °C

213214

Step 4. ^,E-N-Methyl-3-(2-methyl-4-oxo-2,3,4,5-tetrahydro-1H-pyrido[2,3-b][1,4]diazepine-8 -yl)Na((3-methylbenzofuran-2-yl)methyl)acrylamide (compound 214).

The flask is loaded with N)-8-bromo-2-methyl-2,3-dihydro-1H-pyrido[2,3-b][1,4]diazepin-4(5H)-one 213 (80 mg, 0. 31 mmol), Namethyl-N-((3-methylbenzofuran-2-yl)methyl)acrylamide (9) (70 mg, 0.31 mmol), tetrabutylammonium chloride hydrate (9.2 mg, 0.03 mmol) and Pd-116 (16 mg, 0.03 mmol) and the flask was evacuated and backfilled with N ₂ three times. 1,4-dioxane (2.5 ml) and DIPEA (0.11 ml) were added and the reaction mixture was heated to 90°C for 2 hours. Water (10 ml) was then added and the aqueous mixture was extracted.

DXM (3x10 ml). The combined organic layers are dried by passing through a phase separator cartridge and concentrated in vacuo. The crude product was purified by chromatography (0-10% MeOH/DCM) to give the title compound 214 as a yellow solid (68 mg, 52%). R _t 1.89 min (method 1a) m/z 405 [M+H]+ (ER+). ^ChI NMR (DMSO-06, 363 K): δ, ppm, 9.42-9.22 (m, 1I), 8.02 (d, J=1.9 Hz, 1I), 7.56 (dd, J=7.4 Hz, 1.5 Hz, 1I), 7.53-7.38 (m, 3I), 7.27 (dtd, J=20.7 Hz, 7.3 Hz, 1 ,3 Hz, 2I), 7.12 (d, J=15.4 Hz, 1H), 5.57-5.38 (m, 1H), 4.85 (s, 2H), 3.81 (ddt , J=7.7 Hz, 6.3 Hz, 3.2 Hz, 1H), 3.10 (s, 3H), 2.61 (dd, J=14.3 Hz, 3.1 Hz, 1H) , 2.45 (dd, J=14.4 Hz, 7.7 Hz, 1H), 2.28 (s, 3H), 1.25 (d, J=6.4 Hz, 3H).

- 113 046887

Example 46. Synthesis of N,E-3-(3-hydroxy-4-oxo-2,3,4,5-tetrahydro-III-pyrido[2,3-b][1,4]diazepin-8 yl)- N'-methyl-N'-((3-methylbenzofuran-2-yl)methyl)acrylamide (compound 229) and (S,E)-3-(3-rugpoKCu-4octo-2,3,4,5-tetra1hydro -111-11irido[2,3-b][1,4]diaze11in-8-yl)-\-Y1ethyl-\-((3-Y1ethylbenzopho ^; ran-2-yl)\1ethyl)acrylamide (compound 230).

General synthesis scheme.

Reaction conditions: a) MeOH, SOCl ₂ , 0°C to RT; b) CBZ-C1 217, TEA, THF, 0°C; SEM-C1 219, DIPEA, EDC, 60°C; d) Pd/C, H ₂ , 5 bar; e) 5-bromo-3-fluoro-2-nitropyridine 11, TEA, EtOH, reflux; f) Fe, NH ₄ Cl, EtOH, H ₂ O, reflux; h) GATU, DIPEA,

DMF; i) a) TPA, b) Na ₂ COs; j) Chiralpak IA column (25% acetonitrile:75% water); k) 9, Pd-116, DIPEA,

NBu ₄ Cl, 1,4-dioxane, 80°C.

/member Ui MeOH, Social h ₂ n^V ^c ° ^2H -------on o°c-KT

CIH.H ₂ N^Y ^C ° ^2Me

OH

215 216

Step 1. Methyl 3-amino-2-hydroxypropanoate hydrochloride (compound 216).

To a suspension of 3-amino-2-hydroxypropanoic acid 215 (4.0 g, 38 mmol) in methanol (154 ml) stirred in ice water, add thionyl chloride (8.3 ml, 114 mmol) dropwise. After addition is complete, the mixture is stirred at RT overnight. The reaction mixture was evaporated to dryness to obtain the title compound 216 as a colorless gum (6.1 g, quant.). ¹ H NMR (DMSO^): δ, ppm, 8.25 (brs, 3H), 6.07 (brs, 1H), 4.41 (dd, J=8.7 Hz, 3.8 Hz, 1H), 3.68 (s, 3H), 3.12-3.06 (m, 1H), 2.96-2.86 (m, 1H).

Λ _,CO,Me Cbz-ci 217,TEA Cbz. .l ,CO-,Me

ICIH.H ₂ N^< ² ------------ NY ^{2 Z} II—II

OH THF, O °C he

216 218

Step 2. Methyl 3-(((benzyloxy)carbonyl)amino)-2-hydroxypropanoate (compound 218).

To an ice-cold mixture of methyl 3-amino-2-hydroxypropanoate hydrochloride 216 (5.92 g, 38 mmol) and triethylamine (22 ml, 152 mmol) in THF (100 ml) add benzyl chloroformate 217 (5.4 ml, 38) dropwise mmol). The mixture is warmed to RT and stirred for a further 2 hours. The mixture is evaporated to dryness and the residue is redissolved in ethyl acetate (100 ml), washed with water (2x100 ml) and the organic phase is dried over sodium sulfate. The crude product is purified by chromatography on silica

- 114 046887 (0-50% EtOAc/isohexane) to give the title compound 218 as a clear, colorless oil (5.50 g, 56%). ¹ H NMR (CDCl ₃ ): δ, ppm, 7.40-7.28 (m, 5H), 5.15-5.10 (m, 3H), 4.28 (d, J=4, 8 Hz, 1H), 3.78 (s, 3H), 3.66-3.49 (m, 2H), 3.19 (s, 1H).

Cbz^ _N ^-x.CO ₂ Me SEM-CI219, DIPEA, cbz _x ^x\-CO ₂ Me n Iμ I

OH DCE, 60'СOSEM

218220

Step 3. Methyl 3-(((benzyloxy)carbonyl)amino)-2-((2-trimethylsilyl)ethoxy)methoxy)propanoate (compound 220).

Mixture of 2-(trimethylsilyl)ethoxymethyl chloride 219 (2.3 ml, 13.2 mmol), DIPEA (3.4 ml, 19.7 mmol) and methyl 3-(((benzyloxy)carbonyl)amino)-2-hydroxypropanoate 218 (3.30 g, 13.2 mmol) in EDC (100 ml) was stirred at 60°C for 24 hours. The mixture was cooled to rt, the solvent was removed in vacuo, and the crude product was purified by chromatography on silica (0-50% EtOAc/isohexane) to give the title compound 220 as a clear, colorless oil (4.40 g, 83%). ¹ H NMR (CDCl ₃ ): δ, ppm, 7.41-7.32 (m, 5H), 5.19 (brs, 1H), 5.18-5.09 (m, 2H), 4 .78 (s, 2H), 4.29 (dd, J=6.3 Hz, 4.3 Hz, 1H), 3.76 (s, 3H), 3.66 (m, 3H), 3.54 (m, 1H), 0.92 (m, 2H), 0.03 (s, 9H).

Pd/C N

Cbz. l,CO ₂ Me _____' ² ' , /\.CO ₂ Me

N H2N ^H OSEM ^EtOAc OSEM

220 221

Step 4. Methyl 3-amino-2-((2-(trimethylsilyl)ethoxy)methoxy)propanoate (compound 221).

A mixture of methyl 3 -(((benzyloxy)carbonyl)amino)-2-((2-(trimethylsilyl)ethoxy)methoxy)propanoate 220 (4.40 g, 11.5 mmol) and 10% Pd/C (1.2 g, Type 87L) in ethyl acetate solvent (100 ml) was stirred under H2 at 5 bar for 1 hour. The catalyst was removed by filtration and the filtrate was evaporated to dryness to give the title compound 221 as a yellow oil (2.6 g, 89 %). ¹ H NMR (DMSO-dg): δ, ppm, 4.66 (dd, J=13.2 Hz, 4.7 Hz, 2H), 4.01 (dd, J=6.5 Hz, 4 .7 Hz, 1H), 3.64 (s, 3H), 3.63-3.50 (m, 2H), 2.82 (m, 1H), 2.75 (m, 1H), 1.45 (s, 2H), 0.91-0.77 (m, 2H), 0.01 (s, 9H).

_and OSEM

1^. with Me 5-br° ^m -3-fluoro-2-nitropyridine 11, _Br

Η ₂ Ν ^χ η^' ² ------------------------ '' ^x 'oo ₂ me

OSEM TEA, ^E,on · ^kip - °br- COL. P JL

NNO,

221 222

Step 5. Methyl 3-((5-bromo-2-nitropyridin-3-yl)amino)-2-((2-(trimethylsilyl)ethoxy)methoxy)propanoate (compound 222).

A mixture of methyl 3-amino-2-((2-(trimethylsilyl)ethoxy)methoxy)propanoate 221 (2.60 g, 10.4 mmol), TEA (7.3 ml, 52.1 mmol) and 5-bromo- 3-fluoro-2-nitropyridine 11 (2.30 g, 10.4 mmol) in EtOH (80 ml) was heated to reflux for 2 hours. The reaction mixture was evaporated to dryness and the residue was purified by chromatography on silica ( 0-50% EtOAc/isohexane) to give the title compound 222 as a yellow oil (4.30 g, 90%). Rt 2.67 min (method 1a); m/z 448/450 [M+H]+ (ER+). ^1H NMR (DMSO-dg): δ, ppm, 8.08 (t, J=6.1 Hz, 1H), 8.00 (d, J=1.9 Hz, 1H), 7.91 (d, J=1.8 Hz, 1H), 4.76-4.62 (m, 2H), 4.44 (dd, J=7.2 Hz, 4.6 Hz, 1H), 3.77 (kv, J=7.3 Hz, 6.2 Hz, 2H), 3.69 (s, 3H), 3.55-3.43 (m, 2H), 0.81-0.70 (m, 2H), -0.06 (s, 9H).

Step 6. Methyl 3-((2-amino-5-bromopyridin-3-yl)amino)-2-((2-(trimethylsilyl)ethoxy)methoxy)propanoate (compound 223).

Mixture of methyl 3-((5-bromo-2-nitropyridin-3-yl)amino)-2-((2-(trimethylsilyl)ethoxy)methoxy)propanoate 222 (4.30 g, 9.5 mmol), iron powder (2.67 g, 47.7 mmol) and ammonium chloride (5.1 g, 95.0 mmol) in a solvent of ethanol (100 ml) and water (30 ml) are heated and stirred at reflux for 1 hour The mixture is cooled to rt and filtered through a pad of Celite®. The waste filter is evaporated to dryness and the residue is taken up in DCM (130 ml). The organic matter is washed with water (200 ml) and dried over sodium sulfate. Filtration and evaporation gave the title compound 223 as a yellow oil (3.01 g, 74%). Rt 1.76 min (method 1a); m/z 420/422 [M+H]+ (ER+).

223

224

- 115 046887

Step 7. 3-((2-Amino-5-bromopyridin-3-yl)amino)-2-((2-trimethylsilyl)ethoxy)methoxy)propanoic acid (compound 224).

To a stirred solution of methyl 3-((2-amino-5-bromopyridin-3-yl)amino)-2-((2(trimethylsilyl)ethoxy)methoxy)propanoate 223 (3.30 g, 7.85 mmol) in THF (200 ml) and MeOH (50 ml), a solution of LiOH monohydrate (0.23 g, 9.42 mmol) in water (30 ml) was added and the reaction mixture was stirred at RT for 1 hour. The mixture was evaporated to dryness and the residue was taken up in water (30 ml) and pH adjusted to ~pH 5 by adding acetic acid (~1 ml). The solid was collected by filtration, washed with water (4 mL) and dried to give the title compound 224 as a beige solid (2.75 g, 84%). R _t 1.58 min (method 1a); m/z 406/408 [M+H]+ (ER+). 1H NMR (DMSO-dg): δ, ppm, 7.31 (d, J=2.1 Hz, 1H), 6.75 (d, J=2.1 Hz, 1H), 5.75 ( d, J=6.1 Hz, 2H), 5.37 (s, 1H), 4.67-4.64 (m, 2H), 4.22 (m, 1H), 3.55-3.31 (m, 4H), 0.78-0.74 (m, 2H), -0.06 (s, 9H). No acidic proton is observed.

_u OSEM m ₂ H GATU, DIPEA, Vg DMF

OSEM

225

Br

224 nh ₂

Step 8. 8-Bromo-3-((2-(trimethylsilyl)ethoxy)methoxy)-2,3-dihydro-1H-pyrido[2,3-b][1,4]diazepin-4(5H)-one (connection 225).

To a solution of 3-((2-amino-5-bromopyridin-3-yl)amino)-2-((2-(trimethylsilyl)ethoxy)methoxy)propanoic acid 224 (2.75 g, 6.77 mmol) and DIPEA (6.0 ml, 33.8 mmol) in DMF (3 ml) add GATU (3.0 g, 8.12 mmol) and the mixture is stirred at RT for 30 min. The mixture was poured into ice water (30 mL) and the solid was collected and dried to give the title compound 225 as a beige solid (2.50 g, 93%). R _t 2.27 min (method 1b); m/z 388/390 [M+H]+ (ER+). ^1H NMR (DMSO-dg): δ, ppm, 10.05 (s, 1H), 7.71 (s, 1H), 7.28 (s, 1H), 6.21 (d, J= 4.5 Hz, 1H), 4.65 (kv, J=6.9 Hz, 2H), 4.23 (d, J=5.4 Hz, 1H), 3.56-3.45 (m, 4H), 0.88-0.82 (m, 2H), -0.02 (s, 9H).

Step 9. 8-Bromo-3-hydroxy-2,3-dihydro-1H-pyrido[2,3-b][1,4]diazepin-4(5H)-one (compound 226).

TFA (10 ml) was added in one portion to 8-bromo-3-((2-(trimethylsilyl)ethoxy)methoxy)-2,3dihydro-1H-pyrido[2,3-b][1,4]diazepine-4( 5H)-one 225 (800 mg, 2.1 mmol) and the solution was left at RT for 5 min, then carefully poured into crushed ice (100 g). The resulting mixture was carefully neutralized by adding solid sodium carbonate and extracted with ethyl acetate (2x100 ml). The organic substances are combined and dried over sodium sulfate. Filtration and evaporation yielded a gum, which was triturated with acetonitrile (10 ml) to give the title compound 226 as an off-white solid (310 mg, 57%). R _t 1.04 min (method 1a); m/z 258/260 [M+H]+ (ER+). 1H NMR (DMSO-de): δ, ppm, 10.12 (s, 1H), 7.70 (d, J=2.0 Hz, 1H), 7.28 (d, J=2.1 Hz, 1H), 6.37 (dd, J=6.2 Hz, 2.8 Hz, 1H), 5.24 (d, J=4.7 Hz, 1H), 4.16 (dd, J= 8.4 Hz, 4.8 Hz, 3.1 Hz, 1H), 3.45 (ddd, J=12.3 Hz, 6.1 Hz, 3.1 Hz, 1H), 3.29 (td, J=9.1 Hz, 4.5 Hz, 1H).

Step 10. Chiral resolution of racemate 226.

Enantiomers are separated by chiral prep. HPLC using a Chiralpak® IA (Daicel Ltd.) column (2x25 cm), flow rate 13.5 ml min ^-1 , eluting with a mixture of 25% acetonitrile: 75% water, UV

- 116 046887 determination at 254 nm. Samples are loaded onto the column through an in-column dilution pump, pumping acetonitrile (1.5 ml min ^-1 ) throughout the run, resulting in a combined flow rate of 15 ml min ^-1 . Chirality is determined arbitrarily.

(R)-8-Bromo-3-hydroxy-2,3-dihydro-1H-pyrido[2,3-b][1,4]diazepin-4(5H)-one (compound 227). First eluted isomer: Rt 7.98 min (analytical method: Chiralpak® IA column (4.6 mmx25 mm), flow rate 1 ml min ^-1 , eluent: 30% acetonitrile: 70% 10 mmol aqueous ammonium bicarbonate solution).

(S)-8-Bromo-3-hydroxy-2,3-dihydro-1H-pyrido[2,3-b][1,4]diazepin-4(5H)-one (compound 228). Second eluted isomer: Rt 13.09 min (analytical method: Chiralpak® IA column (4.6 mmx25 mm), flow rate 1 ml min ^-1 , eluent: 30% acetonitrile: 70% 10 mmol aqueous ammonium bicarbonate solution).

Stage 11a. (R,E-3-(3-Hydroxy-4-oxo-2,3,4,5-tetrahydro-1H-pyrido[2,3-b][1,4]diazepin-8-yl)-Nmethyl- Na-((3-methylbenzofuran-2-yl)methyl)acrylamide (compound 229).

N)-8-bromo-3-hydroxy-2,3-dihydro-III-pyrido[2,3b][1,4]diazepin-4(5H)-one 227 (50 mg, 0.19) is loaded into the reaction tube mmol), DIPEA (70 µl, 0.39 mmol), tetrabutylammonium chloride hydrate (6 mg, 0.02 mmol), and Pd[P(tBu) ₃ ] ₂ (Pd-116) (10 mg, 0.02 mmol ), and the tube is then rinsed with nitrogen (5 min). Add 1,4-dioxane (5 ml) and N-methyl-N-((3-methylbenzofuran-2-yl)methyl)acrylamide 9 (49 mg, 0.21 mmol), the reaction mixture is again purged with nitrogen (5 min) and then the reaction mixture is heated and stirred at 80°C for 30 minutes. The reaction mixture was cooled to RT, then evaporated to dryness and the crude product was purified by column chromatography (0-10% MeOH/DCM) to give the title compound 229 as a pale yellow solid (61 mg, 77%). Rt 1.77 min (method 1b); m/z 407 [M+H]+ (ER+). ¹ H NMR (DMSO-ph, 363 K): δ, ppm 9.65 (s, 1H), 7.99 (d, J=1.8 Hz, 1H), 7.56 (d, J= 7.5 Hz, 1H), 7.49-7.37 (m, 3H), 7.33-7.21 (m, 2H), 7.13 (d, J=14.6 Hz, 1H), 5.89 (d, J=5.5 Hz, 1H), 4.91-4.77 (m, 3H), 4.22-4.16 (m, 1H), 3.52 (ddd, J= 12.4 Hz, 6.0 Hz, 3.3 Hz, 1H), 3.27 (ddd, J=12.1 Hz, 8.8 Hz, 2.8 Hz, 1H), 3.10 (s, 3H), 2.28 (s, 3H).

Stage 11b. (S,E)-3-(3-Hydroxy-4-oxo-2,3,4,5-tetrahydro-1H-pyrido[2,3-b][1,4]diazepin-8-yl)-Nmethyl -N-((3-methylbenzofuran-2-yl)methyl)acrylamide (compound 230).

N)-8-bromo-3-hydroxy-2,3-dihydro-III-pyrido[2,3b][1,4]diazepin-4(5H)-one 228 (50 mg, 0.19) is loaded into the reaction tube mmol), DIPEA (70 µl, 0.39 mmol), tetrabutylammonium chloride hydrate (6 mg, 0.02 mmol) and Pd[P(tBu) ₃ ] ₂ (Pd-116) (10 mg, 0.02 mmol) , and the tube is then flushed with nitrogen (5 min). Add 1,4-dioxane (5 ml) and N-methyl-N-((3-methylbenzofuran-2-yl)methyl)acrylamide 9 (50 mg, 0.21 mmol), the reaction mixture is again purged with nitrogen (5 min) and then the reaction mixture is heated and stirred at 80°C for 30 minutes. The reaction mixture was cooled to RT, then evaporated to dryness, and the crude product was purified by chromatography on silica (0-10% MeOH/DCM) to give the title compound 230 as a pale yellow solid (58 mg, 73%). Rt 1.76 min (method 1b); m/z 407 [M+H]+ (ER+). 1H NMR (DMSO-dg, 363 K): δ, ppm 9.65 (s, 1H), 7.99 (d, J=1.8 Hz, 1H), 7.56 (d, J=7 .5 Hz, 1H), 7.49-7.36 (m, 3H), 7.27 (dt, J=21.1 Hz, 7.4 Hz, 2H), 7.13 (d, J=15 .7 Hz, 1H), 5.93-5.84 (m, 1H), 4.90-4.80 (m, 3H), 4.18 (dt, J=8.5 Hz, 3.8 Hz , 1H), 3.52 (ddd, J=12.3 Hz, 6.1 Hz, 3.4 Hz, 1H), 3.27 (ddd, J=12.2 Hz, 8.9 Hz, 2, 8 Hz, 1H), 3.10 (s, 3H), 2.28 (s, 3H).

Example 47. Synthesis of dihydrogen phosphate N,E-3-methyl-8-(3-(methyl((3-methylbenzofuran-2yl)methyl)amino)-3-oxoprop-1-en-1-yl)-4-oxo- 2,3,4,5-tetrahydro-1H-pyrido[2,3-b][1,4]diazepin-3-yl (compound 232).

General synthesis scheme.

- 117 046887

Reaction conditions: a) i) bis-(2-cyanoethyl)diisopropylphosphoramidite, 1H-tetrazole, 1 day, RT; ii) 0.1 M iodine solution (water/2,6-lutidine/THF, 1/19/80) at 0°C, 20% v/v. Na ₂ S ₂ O3; b) BSTFA, ACN, Barton's base, 35 min at 0°C, then HCOOH.

Step 1. 2-Cyanoethyl ((R)-3-methyl-8-((E)-3-(methyl((3-methylbenzofuran-2-yl)methyl)amino)-3oxoprop-1-en-1-yl )-4-oxo-2,3,4,5-tetrahydro-1H-pyrido[2,3-b][1,4]diazepin-3-yl)hydrogen phosphate (compound 231).

To a solution of substrate 49 (250 mg, 0.59 mmol, 1 eq.) in dry DCM (5.95 ml) add bis-(2cyanoethyl)diisopropylphosphoramidite (645 mg, 2.38 mmol, 4 eq.) and 1H-tetrazole (5.29 mL, 2.38 mmol, 4 eq., 0.45 M in MeCN) under N2 at RT. The mixture is stirred at RT for 1 day, then partially evaporated. The resulting phosphite is oxidized by treatment with 0.1 M iodine solution (water/2,6-lutidine/THF, 1/19/80) at 0°C until stable iodine staining occurs. The mixture is allowed to return to RT, then after 1 hour the mixture is quenched using sodium thiosulfate 20 wt/wt% with stirring until the iodine color disappears. The aqueous phase is removed and the organic phase is dried over Na ₂ SO ₄ , diluted with MeOH and DCM and filtered. After evaporation, the crude product was purified by elution with 5-100% MeCN (0.1% TFA) in water (0.1% TFA) to obtain, after lyophilization, the desired product 231 (165 mg, 0.30 mmol, 50%) in as a yellow solid. The product is used immediately in the next stage.

Stage 2. Dihydrogen phosphate ^,E-3-methyl-8-(3-(methyl((3-methylbenzofuran-2-yl)methyl)amino)-3oxoprop-1-en-1-yl)-4-oxo-2 ,3,4,5-tetrahydro-1H-pyrido[2,3-b][1,4]diazepin-3-yl (compound 232).

To a solution of 231 (2.65 g, 4.79 mmol, 1 eq.) in MeCN (28.5 mL) add N,O-bis(trimethylsilyl)trifluoroacetamide (BSTFA) (6.36 mL, 23.9 mmol, 5 equiv.), then 2-tert-butyl-1,1,3,3-tetramethylguanidine (Barton's base) (4.14 ml, 19.9 mmol, 4.1 equiv.). Stir at 0°C for 35 minutes, then add formic acid to quench the reaction mixture. After evaporation, the crude product was purified by eluting with 5-100% MeCN (0.1% HCO2H) in water (0.1% HCO2H) and lyophilized to give the desired compound 232 (1.26 g, 250 mmol, 52%). ^1H NMR (400 MHz, DMSO-de): δ, ppm 9.89 (s, 1H), 8.05-7.96 (m, 1H), 7.61-7.53 (m, 1H ), 7.52-7.02 (m, 6H), 6.20 (m, 1H), 4.95 (s, 0.8H), 4.79 (s, 1.2H), 3.62- 3.41 (m, 2H), 3.17 (s, 2H), 2.95 (s, 1H), 2.31-2.21 (s, 3H). ³¹ P NMR (162 MHz, DMSO-06) δ -4.8. MS (method 1c): m/z=501.1 [M+H]+.

Example 48. Synthesis of [(2R,3S)-2-methyl-8-[(E)-3-[methyl-[(3-methylbenzofuran-2-yl)methyl]amino]-3oxoprop-1-enyl]-4 -oxo-1,2,3,5-tetrahydropyrido[2,3-b][1,4]diαzepin-3-yl]dihydrogen phosphate - salt (2R,3R,4R, 5S)-6-(methylamino)hexane- 1,2,3,4,5-pentol (compound 235).

General synthesis scheme.

150 233

234 235

Reaction conditions: a) i) bis-(2-cyanoethyl)diisopropylphosphoramidite, III-tetrazole, 1 day, RT; ii) 0.1 M iodine solution (water/2,6-lutidine/THF, 1/19/80) at 0°C, 20% v/v. Na ₂ S ₂ O ₃ ; b) BSTFA, ACN, 2-tert-butyl-1,1,3,3-tetramethylguanidine, 35 min at 0°C, then HCOOH; c) H ₂ O and MeCN (60 ml, 1/1), meglumine 2 eq.

Step 1. 2-Cyanoethyl ((2R,3S)-2-methyl-8-((E)-3-(methyl((3-methylbenzofuran-2-yl)methyl)amino)-3-oxoprop1-ene-1 -yl)-4-oxo-2,3,4,5-tetrahydro-1H-pyrido[2,3-b][1,4]diazepin-3-yl)hydrogen phosphate (compound 233).

To mixture 150 (129 mg, 0.31 mmol, 1 eq.) in dry CH ₂ Cl ₂ (2.80 ml) add bis-(2cyanoethyl)diisopropylphosphoramidite (0.24 ml, 0.92 mmol, 3 eq.) and 1H-tetrazole (1.36 mL, 0.61 mmol, 2 eq., 0.45 M in MeCN). The mixture is stirred at RT for 16 hours. After completion of the reaction, 0.2 M iodine solution (I ₂ in water/pyridine/THF, 1/19/80) is added at 0°C until the iodine color is stable. The mixture is allowed to return to RT, then after 2 hours the mixture is quenched using sodium thiosulfate (20 wt/wt%) with stirring until the iodine color disappears. The aqueous phase is removed and the organic phase is dried over Na ₂ SO ₄ , diluted with MeOH and CH ₂ Cl ₂ and filtered. After evaporation, the crude product was purified by elution with 5-100% MeCN (0.1% TFA) in water (0.1% TFA) to obtain, after lyophilization, the desired compound 233 (159 mg) as a yellow solid. The product is used immediately in the next stage.

Stage 2. Dihydrogen phosphate (2R,3S)-2-Methyl-8-((E)-3-(methyl((3-methylbenzofuran-2yl)methyl)amino)-3-oxoprop-1-en-1-yl) -4-oxo-2,3,4,5-tetrahydro-1H-pyrido[2,3-b][1,4]diazepin-3-yl (compound 234).

To a mixture of 233 (159 mg, 0.29 mmol, 1 eq.) in dry MeCN (14.4 ml) add N,O-bis

- 118 046887 (trimethylsilyl)trifluoroacetamide (0.31 ml, 1.15 mmol, 4 eq.), then 2-tert-butyl-1,1,3,3tetramethylguanidine (0.20 ml, 0.96 mmol, 3. 3 eq.). The mixture was stirred at 0°C for 10 min, then formic acid (0.11 ml, 2.87 mmol, 10 eq.) was added to quench the reaction mixture. After evaporation, the crude product was purified by eluting with 5-100% MeCN (0.1% HCO ₂ H) in water (0.1% TFA) to obtain, after lyophilization, the desired compound 234 (33 mg, 0.06 mmol, 22 %) as a yellow solid. The desired compound is used to prepare the meglumine salt.

Step 3. [(2R,3S)-2-Methyl-8-[(E)-3-[methyl-[(3-methylbenzofuran-2-yl)methyl]amino]-3-oxoprop-1 enyl]-4 -oxo-1,2,3,5-tetrahydropyrido[2,3-b][1,4]diαzepin-3-yl]dihydrogen phosphate - (2R,3R,4R,5S)-6(methylamino)hexane-1, 2,3,4,5-pentol salt (compound 235).

To a solution of substrate 234 (1 equiv.) in water and MeCN (60 ml, 1/1) add the appropriate base (meglumine, 2 equiv.) at RT. The mixture is stirred, sonicated and lyophilized to give the desired product 235. ¹ H NMR (400 MHz, D2O): δ, ppm 7.80-7.65 (m, 1H), 7.25-7.46 (m , 7H), 4.52-4.25 (m, 2H), 4.18-4.02 (m, 3H), 3.83-3.69 (m, 6H), 3.67-3.56 (m, 4H), 3.26-3.04 (m, 4H), 2.89-2.75 (m, 6H), 2.72 (s, 6H), 1.84 (s, 3H), 1.25-1.16 (m, 6H). ³¹ P NMR (162 MHz, D2O) δ 3.1. MS (method 1c): m/z=501.1 [M+H]+.

Example 49. Synthesis of 2-aminoethanol; dihydrogen phosphate (I,E-8-(3-(((7-amino-2-methylbenzofuran-3yl)methyl)(methyl)amino)-3-oxoprop-1-en-1-yl)-3-methyl-4 -oxo-2,3,4,5-τtetrahydro-1H-pyrido[2,3-b][1,4]diazepin-3yl (compound 237).

General synthesis scheme.

237

Reaction conditions: a) i) bis-(2-cyanoethyl)diisopropylphosphoramidite, III-tetrazole, 1 day, RT; ii) 0.1 M iodine solution (water/2,6-lutidine/THF, 1/19/80) at 0°C, 20% v/v. Na ₂ S ₂ O ₃ ; b) i) BSTFA, ACN, Barton's base, 35 min at 0°C, then HCOOH; ii) water, HOCH2CH2NH2 (5.0 eq.)

Step 1. 2-Cyanoethylhydro (3-(((E)-3-((3R)-3-(((2-cyanoethoxy)(hydroxy)phosphoryl)oxy)-3methyl-4-oxo-2,3,4 ,5-tetrahydro-1H-pyrido[2,3-b][1,4]diazepin-8-yl)-N-methylacrylamido)methyl)-2-methylbenzofuran-7-yl)phosphoramidate (compound 236).

To mixture 167 (200 mg, 0.46 mmol, 1 eq.) in dry CH ₂ Cl ₂ (4.59 ml) add bis-(2cyanoethyl)diichopropylphosphoramidite (0.48 ml, 1.84 mmol, 4 eq.) and N-tetrazole (4.08 ml, 1.84 mmol, 4 eq., 0.45 M in MeCN). The mixture is stirred at RT for 16 hours, then partially evaporated, and 0.2 M iodine solution (water/pyridine/THF, 1/19/80) is added at 0°C until the iodine color is permanent. The mixtures allow you to return to the CT scan. After 2 hours, the mixture is quenched using sodium thiosulfate 20 wt./wt.% with stirring until the iodine color disappears. The aqueous phase is removed and the organic phase is dried over Na ₂ SO ₄ , diluted with MeOH and CH ₂ Cl ₂ and filtered. After evaporation, the crude product was purified by elution with 5-100% MeCN (0.1% TFA) in water (0.1% TFA) to obtain, after lyophilization, the desired compound 236 (245 mg) as a yellow solid.

Stage 2. 2-Aminoethanol dihydrogen phosphate ^,E-8-(3-(((7-amino-2-methylbenzofuran-3-yl)methyl)(methyl)amino)-3-oxoprop-1-en-1-yl )-3-methyl-4-oxo-2,3,4,5-tetrahydro-1H-pyrido[2,3-b][1,4]diazepin-3yl (compound 237).

To a mixture of 236 (240 mg, 0.34 mmol, 1 eq.) in dry MeCN (18.9 mL) add BSTFA (0.91 mL, 3.42 mmol, 10 eq.), then 2-tert-butyl- 1,1,3,3-tetramethylgcanidine (Barton's base) (0.59 mL, 2.85 mmol, 8.3 eq.). The mixture is stirred at 0°C for 5/10 min, then HCOOH is added to quench the reaction mixture and water (1 ml). After evaporation, the crude product is purified by elution with 5-100% MeCN (0.1% TFA) in water (0.1% TFA) to obtain a powder containing the desired dihydrogen phosphate. The residue is dissolved in water+ethanolamine in excess (>5 eq.). The mixture was purified by eluting with 5-100% MeCN in water to obtain, after lyophilization, the desired compound 237 (89 mg, 0.15 mmol, 45%) as a yellow solid. 1H NMR (400 MHz, D2O): δ, ppm 7.88-7.78 (m, 1H), 7.44-7.26 (m, 2H), 7.21-6.61 (m, 4H), 4.76-4.59 (m, 2H), 3.59 (t, J=5.5 Hz, 1H), 3.55-3.46 (m, 2H), 3.00-2 .81 (m, 3H), 2.77 (t, J=5.5 Hz, 1H), 2.46-2.28 (m, 3H), 1.59-1.46 (m, 3H). ³¹ P NMR (162 MHz, D2O) δ 0.0. MS (method 1c): m/z=516.1 [M+H] ⁺ .

Example 50. Synthesis of dihydrogen phosphate (R)-3-methyl-4-oxo-8-((E)-3-oxo-3-((3aS,6aR)-5-(4- 119 046887 phenoxyphenyl)-3,3a ,4,6a-tetrahydrocyclopenta[c]pyrrol-2(1H)-yl)prop-1-en-1-yl)-2,3,4,5-tetrahydro1H-pyrido[2,3-b][1, 4]diazepin-3-yl (compound 239).

General synthesis scheme.

Reaction conditions: a) i) bis-(2-cyanoethyl)diisopropylphosphoramidite, III-tetrazole, 1 day, RT; ii) 0.1 M iodine solution (water/2,6-lutidine/THF, 1/19/80) at 0°C, 20% v/v. Na ₂ S ₂ O ₃ ; b) BSTFA, ACN, Barton's base, 35 min at 0°C, then HCOOH.

Step 1. 2-Cyanoethyl ((R)-3-Methyl-4-oxo-8-((E)-3-oxo-3-((3aS,6aR)-5-(4-phenoxyphenyl)3,3a, 4,6a-tetrahydrocyclopenta[c]pyrrol-2(1H)-yl)prop-1-en-1-yl)-2,3,4,5-tetrahydro-1H-pyrido[2,3b][1,4 ]diazepin-3-yl)hydrogen phosphate (compound 238).

To mixture 159 (400 mg, 0.77 mmol, 1 eq.) in dry CH ₂ Cl ₂ (7.65 ml) add bis-(2cyanoethyl)diichopropylphosphoramidite (831 mg, 3.06 mmol, 4 eq.) and III -tetrazole (6.80 ml, 3.07 mmol, 4 eq., 0.45 M in MeCN). The mixture is stirred at RT for 16 hours, then evaporated and 0.2 M iodine solution (water/pyridine/THF, 1/19/80) is added at 0°C until the iodine color is stable. The mixture is allowed to return to RT, then after 2 hours the mixture is quenched using aqueous sodium thiosulfate 20 wt/wt% with stirring until the iodine color disappears. The aqueous phase is removed and the organic phase is dried over Na ₂ SO ₄ , diluted with MeOH and CH ₂ Cl ₂ and filtered. After evaporation, the crude product was purified by elution with 5-100% MeCN (0.1% TFA) in water (0.1% TFA) to obtain the desired compound 238 (421 mg) as a yellow solid. The product is used directly in the next step.

Step 2. Dihydrogen phosphate (R)-3-Methyl-4-oxo-8-((E)-3-oxo-3-((3aS,6aR)-5-(4-phenoxyphenyl)3,3a,4,6a -tetrahydrocyclopenta[c]pyrrol-2(1H)-yl)prop-1-en-1-yl)-2,3,4,5-tetrahydro-1H-pyrido[2,3b][1,4]diazepine- 3-yl (compound 239).

To a mixture of 238 (421 mg, 0.64 mmol, 1 eq.) in dry MeCN (35.54 ml) add BSTFA (1.71 ml, 6.42 mmol, 10 eq.), then Barton's base (1.11 ml, 5.35 mmol, 8.3 eq.). The mixture is stirred at RT for 5/10 min, then formic acid is added to quench the reaction mixture. After evaporation, the residue is purified by eluting with 5-100% MeCN (0.1% TFA) in water (0.1% TFA). The precipitate was reduced and lyophilized to give the desired compound 239 (123 mg, 0.20 mmol, 32%) as a yellow solid. ¹ H NMR (400 MHz, DMSO-dg): δ, ppm 9.89 (d, J=5.6 Hz, 1H), 8.11-7.90 (m, 1H), 7.59- 7.27 (m, 7H), 7.15 (t, J=7.4 Hz, 1H), 7.09-6.93 (m, 5H), 6.86 (dd, J=15.5, 6.4 Hz, 1H), 6.13 (s, 1H), 4.15-2.78 (m, 9H), 2.62 (d, J=15.9 Hz, 1H), 1.61- 1.49 (m, 3H). ¹⁹ F NMR (376 MHz, DMSO^6) δ -73.6 (residual traces of TPA). ³¹ P NMR (162 MHz, DMSO^6) δ -4.8. MS (method 1c): m/z=603.2 [M+H]+.

Example 51. Synthesis of 2,2,2-trifluoroacetate dihydrogen phosphate N,E)-8-(3-(methyl((3-methylbenzofuran-2yl)methyl)amino)-3-oxoprop-1-en-1-yl)- 4-oxo-2,3,4,5-tetrahydro-1H-pyrido[2,3-b][1,4]diazepin-3-yl (compound 241).

General synthesis scheme.

Reaction conditions: a) i) bis-(2-cyanoethyl)diisopropylphosphoramidite, III-tetrazole, 1 day, RT; ii) 0.1 M iodine solution (water/2,6-lutidine/THF, 1/19/80) at 0°C, 20% v/v. Na ₂ S ₂ O ₃ ; b) BSTFA, ACN, Barton's base, 35 min at 0°C, then HCOOH.

Stage 1. Hydrogen phosphate 2-cyanoethyl ((S)-8-((E)-3-(methyl((3-methylbenzofuran-2-yl)methyl)amino)-3oxoprop-1-en-1-yl)-4 -oxo-2,3,4,5-tetrahydro-III-pyrido[2,3-b][1,4]diazepin-3-yl) (compound 240). TO

- 120 046887 mixture 230 (200 mg, 0.49 mmol, 1 eq.) in dry DMF (4.92 ml) add bis-(2cyanoethyl)diichopropylphosphoramidite (0.51 ml, 1.97 mmol, 4 eq.) and N-tetrazole (4.37 ml, 1.97 mmol, 4 eq., 0.45 M in MeCN). The mixture is stirred at 60°C for 4 hours, then MeCN is evaporated and 0.2 M iodine solution (water/pyridine/THF, 1/19/80) is added at 0°C until the iodine color is permanent. The mixture is allowed to return to RT, then after 2 hours the mixture is quenched using sodium thiosulfate 20 wt/wt% with stirring until the iodine color disappears. The aqueous phase is removed and the organic phase is dried over Na ₂ SO ₄ , diluted with MeOH and CH ₂ Cl ₂ and filtered. After evaporation, the crude product is purified by elution with 5-100% MeCN (0.1% TFA) in water (0.1% TFA) to obtain the desired compound 240 (199 mg) after lyophilization. The product is used directly in the next step.

Step 2. 2,2,2-Trifluoroacetate dihydrogen phosphate ^,E)-8-(3-(Methyl((3-methylbenzofuran-2yl)methyl)amino)-3-oxoprop-1-en-1-it)-4 -oxo-2,3,4,5-tetrahydro-1H-pyrido[2,3-b][1,4]diazepin-3-yl (compound 241). To a mixture of 240 (120 mg, 0.22 mmol, 1 eq.) in dry MeCN (12.3 mL) add BSTFA (1.71 mL, 6.42 mmol, 10 eq.). The mixture was stirred at RT for 10 min, then formic acid (0.08 ml, 2.22 mmol, 10 eq.) was added to quench the reaction mixture and MeOH (1 ml). After evaporation, the crude product is purified eluting with 5-100% MeCN (0.1% TFA) in water (0.1% TFA) to obtain, after lyophilization, the desired compound (42 mg, 0.07 mmol, 31%) in as a yellow solid. ¹ H NMR (400 MHz, DMSO-d^: δ, ppm 10.23 (s, 1H), 8.18-8.04 (m, 1H), 7.57 (d, J=7.4 Hz, 1H), 7.54-7.35 (m, 4H), 7.31-7.07 (m, 3H), 6.10 (s, 1H), 4.96 (s, 0.8H) , 4.80 (s, 1.2H), 4.74-4.57 (m, 1H), 3.69 (dd, J=12.3, 3.3 Hz, 1H), 3.51 (dd , J=12.3, 8.5 Hz, 1H), 3.18 (s, 1.7H), 2.95 (s, 1.3H), 2.28 (d, J=4.4 Hz, ^{3H )} . ,0 [M+H]+.

Example 52. Synthesis of ^,E-3-(6-hydroxy-6-methyl-7-oxo-5,6,7,8-tetrahydro-1,8-naphthyridin-3-yl)-N methyl-^(( 3-methylbenzofuran-2-yl)methyl)acrylamide (compound 247).

General synthesis scheme.

Reaction conditions: a) \1,\1,\'1,\'1,\2,\2,\'2,\'2-octamethyl..t)ten-1,1,2,2-hetraami11, DMF -20°C to RT; b) Fe, NH4C1, EtOH, water, 90°C; c) NBS, K ₂ CO ₃ , MeCN, then chiral separation; d) Pd-116, DIPEA, NBu ₄ Cl, 1,4-dioxane, 80°C.

Step 1. Ethyl 2-hydroxy-2-methyl-3-(2-nitropyridin-3-yl)propanoate (compound 244).

To a stirred solution of 3-(bromomethyl)-2-nitropyridine 242 (850 mg, 3.92 mmol) in anhydrous DMF (15 ml), add ethyl pyruvate 243 (1.3 ml, 11.8 mmol) at -20°C. The resulting solution is stirred and kept at this temperature for 30 minutes. M,M,#1,#1,#,#,#2,#2-octamethylethene-1,1,2,2-tetraamine (1.0 mL, 4.31 mmol) was then added. A red color appears immediately with the formation of a fine white precipitate. The solution is stirred vigorously at -20°C for 1 hour and then warmed to RT for 2 hours. The orange-red cloudy solution is filtered and water (80 ml) is added. The aqueous layer is extracted with ethyl acetate (3x40 ml) and the combined organic extracts are washed with water (3x40 ml), dried over MgSO ₄ , filtered and concentrated in vacuo. The crude product was purified by chromatography (0-10% MeOH/DCM) to give the title compound 244 as a colorless solid (630 mg, 57%). R _t 1.48 min (method 1a) m/z 255 [M+H]+ (ER+).

Step 2. 3-Hydroxy-3-methyl-3,4-dihydro-1,8-naphthyridin-2(III)-one (compound 245).

Iron ( 554 mg, 9.9 mmol) and ammonium chloride (530 mg, 9.91 mmol). The resulting mixture is heated at reflux for 1 hour. The reaction mixture is filtered through a pad of Celite® while it is hot. The filtrate is then evaporated to dryness. The crude product was purified by chromatography (0-10% MeOH/DCM) to give the title compound 245 as a colorless solid (350 mg, 75%). R _t 0.56 min (method 1a) m/z 179 [M+H]+ (ER+).

Step 3. ^)-6-Bromo-3-hydroxy-3-methyl-3,4-dihydro-1,8-naphthyridin-2(III)-one (compound 246).

N-Bromosuccinimide (619 mg, 3.48 mmol) and K2CO3 (962 mg, 6.96 mmol) were added in one portion to

- 121 046887 stirred solution of 3-hydroxy-3-methyl-3,4-dihydro-1,8-naphthyridin-2(1H)-one 245 (310 mg, 1.74 mmol) in acetonitrile (20 ml) at 0° C. The reaction mixture is warmed to RT and then stirred overnight. After this, more N-bromosuccinimide (619 mg, 3.48 mmol) and K ₂ CO ₃ (962 mg, 6.96 mmol) were added and the resulting mixture was stirred for 3 hours. The reaction mixture was quenched by adding H2O (10 ml), then extract with DCM (3x10 ml). The combined organic layers are dried by passing through a phase separator cartridge and concentrated in vacuo. The crude product was purified by chromatography (0-100% EtOAc/isohexane) to give the racemate as a white solid (200 mg, 42%). The racemate was subjected to chiral resolution to obtain the first peak (Q)-6-bromo-3-hydroxy-3methyl-3,4-dihydro-1,8-naphthyridin-2(III)-one 246 (60 mg). Rt 1.19 min (method 1a) m/z 257/259 [M+H]+ (ER+). The absolute configuration is determined arbitrarily.

Stage 4. U,E-3-(6-Hydroxy-6-methyl-7-oxo-5,6,7,8-tetrahydro-1,8-naphthyridin-3-yl)-U-methyl-U(( 3-methylbenzofuran-2-yl)methyl)acrylamide (compound 247).

N)-6-bromo-3-hydroxy-3-methyl-3,4-dihydro-1,8-naphthyridin-2(III)-one 246 (60 mg, 0.23 mmol), Nmethyl- Na-((3-methylbenzofuran-2-yl)methyl)acrylamide 9 (54 mg, 0.23 mmol), NBu ₄ Cl (7 mg, 0.03 mmol) and Pd-116 (12 mg, 0.03 mmol ), and the flask is evacuated and backfilled with N2 three times. 1,4-Dioxane (2 ml) and DIPEA (85 µl, 0.46 mmol) were added and the reaction mixture was heated to 80°C for 2 hours. Water (2 ml) was then added and the aqueous mixture was extracted with DCM (3x5 ml). The combined organic extracts are dried by passing through a phase separator cartridge and concentrated in vacuo. The crude product is purified by column chromatography (0-5% MeOH/DCM). The resulting product is triturated with MeCN (2 ml). The precipitate was collected by filtration and dried to give the title compound 247 as a pale yellow solid (30 mg, 32%). Rt 1.83 min (method 1a) m/z 406 [M+H]+ (ER+). ^1H NMR (DMSO-dg): δ, ppm, 10.29 (s, 1H), 8.36 (d, J=2.2 Hz, 1H), 7.97 (d, J=2, 1 Hz, 1H), 7.61-7.43 (m, 3H), 7.34-7.14 (m, 3H), 5.19 (d, J=1.5 Hz, 1H), 4, 86 (s, 2H), 3.11 (s, 3H), 2.94 (s, 2H), 2.28 (s, 3H), 1.29 (s, 3H).

Example 53. Synthesis of (E)-3-(6-ethyl-6-hydroxy-7-oxo-5,6,7,8-tetrahydro-1,8-naphthyridin-3-yl)-Nmethyl-N-(( 3-methylbenzofuran-2-yl)methyl)acrylamide (compound 252).

General synthesis scheme.

²⁵¹ 252

Reaction conditions: a) M,III,No.1,No.1,No.,No.,No.2,No.2-octamethylethene-1,1,2,2-tetraamine, DMF -20°C to RT; b) Fe, NH ₄ Cl, EtOH, water, 90°C; c) NBS, K ₂ CO ₃ , ACN, 0°C to RT; d) Pd-116, DIPEA, NBu ₄ Cl, 1,4dioxane, 80°C.

Step 1. Ethyl 2-hydroxy-2-ethyl-3-(2-nitropyridin-3-yl)propanoate (compound 249).

To a stirred solution of 3-(bromomethyl)-2-nitropyridine 242 (425 mg, 1.96 mmol) in anhydrous DMF (7.5 ml), ethyl 2-oxobutanoate 248 (765 mg, 5.88 mmol) at -20° C. The resulting solution is stirred for 30 minutes. Then add ^^1,#1,#1,#,#,#2,#2-octamethylethene-1,1,2,2tetraamine (0.5 ml, 2.15 mmol). The solution is stirred vigorously at -20°C for 1 hour and then warmed to RT for 2 hours. The orange-red cloudy solution is filtered and hydrolyzed with water (40 ml). The aqueous layer was extracted with ethyl acetate (3x20 ml), the combined organic layers were washed with water (3x20 ml), dried over MgSO ₄ , filtered and concentrated in vacuo. The crude product was purified by chromatography (0-10% MeOH/DCM) to give the title compound 249 as a colorless solid (380 mg, 66%). Rt 1.75 min (method 1a) m/z 269 [M+H]+ (ER+).

Step 2. 3-Hydroxy-3-ethyl-3,4-dihydro-1,8-naphthyridin-2(III)-one (compound 250). To a stirred solution of ethyl 2-hydroxy-2-((2-nitropyridin-3-yl)methyl)butanoate 249 (370 mg, 1.38 mmol) in ethanol (18 ml) and water (5 ml) add iron (308 mg , 5.52 mmol) and ammonia hydrochloride (295 mg, 5.52 mmol). The reaction mixture is heated at 90°C for 1 hour. The reaction mixture is cooled to room temperature and filtered through a pad of Celite®. The filtrate was evaporated to dryness and the crude product was purified by chromatography (0-10% MeOH/DCM) to give the title compound 250 as a colorless solid (200 mg, 72%). Rt 0.89 min (method 1a) m/z 193 [M+H]+ (er ⁺ ).

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Step 3. 6-Bromo-3-hydroxy-3-ethyl-3,4-dihydro-1,8-naphthyridin-2(1H)-one (compound 251).

N-Bromosuccinimide (361 mg, 2.03 mmol) and K2CO3 (561 mg, 4.06 mmol) were added in one portion to the stirred solution of 3-ethyl-3-hydroxy-3,4-dihydro-1,8-naphthyridine-2 (1H)-one 250 (195 mg, 1.01 mmol) in acetonitrile (15 ml) at 0°C. The reaction mixture is returned to RT and stirred overnight. The reaction mixture is filtered over a pad of celite and the solids are washed with acetonitrile (20 ml). The filtrate was evaporated in vacuo and the crude product was purified by chromatography (0-100% EtOAc/isohexane) to give the title compound 251 as a white solid (142 mg, 51%). R _t 1.20 min (method 2a) m/z 271/273 [M+H]+ (ER+). ¹ H NMR (400 MHz, CDCl ₃ ): δ, ppm 9.21 (s, 1H), 8.32 (dd, J=2.3 Hz, 1.1 Hz, 1H), 7.69 ( t, J=1.7 Hz, 1H), 3.72 (s, 1H), 3.14 (d, J=16.1 Hz, 1H), 3.07 (d, J=16.1 Hz, 1H), 1.68-1.50 (m, 2H), 0.96 (t, J=7.4 Hz, 3H).

Step 4. (E)-3-(6-Ethyl-6-hydroxy-7-oxo-5,6,7,8-tetrahydro-1,8-naphthyridin-3-yl)-N-methyl-N-( (3methylbenzofuran-2-yl)methyl)acrylamide (compound 252).

Namethyl-Na((3-methylbenzofuran-2-yl)methyl)acrylamide 9 (118 mg, 0.52 mmol), 6-bromo-3-ethyl-3-hydroxy-3,4-dihydro- 1,8-naphthyridin-2(1H)-one 251 (140 mg, 0.52 mmol), tetrabutylammonium chloride hydrate (15 mg, 0.05 mmol) and Pd-116 (26 mg, 0.05 mmol) and test tube then washed with nitrogen (5 min). 1,4-Dioxane (8 mL) and DIPEA (180 μL, 1.03 mmol) are added to the tube and the reaction mixture is heated to 80°C for 50 min. The crude product is evaporated in vacuo and purified by chromatography (0-10% MeOH/DCM). The product was recrystallized in DCM/hexane (5 mL) then triturated with acetonitrile (2 mL) to give the title compound 252 as a white solid (144 mg, 66%). Rt 1.59 min (method 2a) m/z 420 [M+H]+ (ER+). 1H NMR (400 MHz, DMSO-de, 363 K): δ, ppm, 10.33 (s, 1H), 8.35 (d, J=2.1 Hz, 1H), 8.01-7 .97 (m, 1H), 7.61-7.53 (m, 1H), 7.53-7.44 (m, 2H), 7.32-7.20 (m, 3H), 4.95 (s, 1H), 4.86 (s, 2H), 3.11 (s, 3H), 2.97 (s, 2H), 2.28 (s, 3H), 1.66-1.48 ( m, 2H), 0.89 (t, J=7.4 Hz, 3H).

Example 54. Synthesis of (R,E-3-(7-hydroxy-7-methyl-8-oxo-8,9-dihydro-7H-pyrido[2,3-b]azepin-3-yl)Nmethyl-N ((3-methylbenzofuran-2-yl)methyl)acrylamide (compound 260).

General synthesis scheme.

bromide

b)

Conditions before CT;

bireaction chloride: a) ethynylmagnesium, -78°C (triphenylphosphine)palladium(P), 5-bromo-3-iodopyridyl-2-amine 255, CuI, TEA, THF, 80°C; c) H ₂ O, Pd/C,

EtOAc; d) NaH, THF, 0°C to RT; e) Pd-116, DIPEA, NBu ₄ Cl, 1,4-dioxane, 80°C; f) chiral separation.

Step 1. Ethyl 2-hydroxy-2-methylbut-3-inoate (compound 254).

A solution of ethyl magnesium bromide (0.5 M in THF, 99 mL, 49.6 mmol) was added dropwise over ~1 h to a solution of ethyl 2-oxopropanoate 253 (4.6 mL, 41.3 mmol) in THF ( 60 ml) at -78°C. The mixture is stirred at this temperature for another 15 minutes, then heated to RT. The reaction mixture was then poured into saturated ammonium chloride solution (50 ml) and the mixture was extracted with TBME (2x30 ml). The combined organic extracts are dried over sodium sulfate, concentrated in vacuo and the crude product is purified by chromatography on silica (0-40% MTBE/isohexane) to give the title compound 254 as a pale yellow oil (0.85 g, 14%) . ¹ H NMR (400 MHz,

DMSO-de): δ, ppm 6.28 (s, 1H), 4.15 (kvd, J=7.1 Hz, 1.5 Hz, 2H), 3.46 (s, 1H), 1 .51 (s, 3H), 1.21 (t, J=7.1 Hz, 3H).

Step 2. Ethyl 4-(2-amino-5-bromopyridin-3-yl)-2-hydroxy-2-methylbut-3-inoate (compound 256).

Copper iodide (31 mg, 0.16 mmol), bis-(triphenylphosphine) palladium chloride (94 mg, 0.13 mmol) and 5-bromo-3-iodopyridin-2-amine 255 (0.80 g, 2.7 mmol), and the tube is evacuated and backfilled with N ₂ three times. Ethyl 2-hydroxy-2-methylbut-3-inoate 254 (0.42 g, 2.95 mmol), TEA (1.1 mL, 8.1 mmol) and THF (10 mL) were added and the mixture was degassed with N2 for 10 minutes, then reflux for 2 hours. The reaction mixture is cooled to rt, filtered through a pad of Celite® and washed with EtOAc (20 ml). The filtrate was concentrated in vacuo and the crude product was purified by silica chromatography (0-100% EtOAc/isohexane) to give the title compound 256 as a clear yellow oil (810 mg, 91%). Rt 1.66 min (method 1a); m/z 313/315 [M+H]+ (ER+). 1H NMR (400 MHz, DMSO-de): δ, ppm 8.03 (d, J=2.4 Hz, 1H), 7.66 (d, J=2.4 Hz, 1H),

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6.57 (s, 1H), 6.45 (s, 2H), 4.19 (kv, J=7.1 Hz, 2H), 1.61 (s, 3H), 1.23 (t, J =7.1 Hz, 3H).

Step 3. (2)-Ethyl 4-(2-amino-5-bromopyridin-3-yl)-2-hydroxy-2-methylbut-3-enoate (compound 257).

A mixture of ethyl 4-(2-amino-5-bromopyridin-3-yl)-2-hydroxy-2-methylbut-3-inoate 256 (0.45 g, 1.44 mmol) and 5% Pd/C (0. 15 g, 1.44 mmol) in EtOAc (50 ml) is stirred under H ₂ at 1 bar for 30 minutes. The catalyst was removed by filtration, then the filtrate was concentrated in vacuo and purified by column chromatography (0-100% EtOAc/isohexane) to give the desired product 257 as a colorless oil (0.17 g, 27%). Rt 0.98 min (method 1a); m/z 315/317 [M+H]+ (ER+). ¹ H NMR (400 MHz, DMSO-de): δ, ppm 7.87 (d, J=2.5 Hz, 1H), 7.66 (d, J=2.5 Hz, 1H), 6 .21 (d, J=12.1 Hz, 1H), 5.97 (s, 2H), 5.81 (d, J=12.1 Hz, 1H), 5.74 (s, 1H), 3 .83-3.68 (m, 2H), 1.41 (s, 3H), 1.03 (t, J=7.1 Hz, 3H).

Step 4. 3-Bromo-7-hydroxy-7-methyl-7H-pyrigo[2,3-b]azepin-8(9H)-one (compound 258).

Sodium hydride (60% in mineral oil, 0.03 g, 0.79 mmol) is added in small portions to a stirred solution of (2)-ethyl 4-(2-amino-5-bromopyridin-3-yl)-2-hydroxy- 2-methylbut-3-enoate 257 (0.17 g, 0.52 mmol) in THF (15 ml) at 0°C. The reaction mixture is heated to RT and stirred for 2 hours. The reaction mixture is quenched by careful addition of sat. aq. NH ₄ Cl (50 ml) and the aqueous mixture was extracted with EtOAc (3x100 ml). The combined organic layers were washed with brine (1x100 mL), dried with MgSO ₄ and concentrated in vacuo to give the desired product 258 as an off-white solid (0.17 g, quant.) which was used in the next step without further purification. R _t 1.38 min (method 1a); m/z 269/271 [M+H]+ (ER+). 1H NMR (400 MHz, DMSOD ₆ ): δ, ppm 10.93 (s, 1H), 8.42 (d, J=2.4 Hz, 1H), 8.10 (d, J=2, 4 Hz, 1H), 6.73 (d, J=10.9 Hz, 1H), 6.04 (d, J=10.9 Hz, 1H), 5.39 (s, 1H), 1.23 (c, 3H).

Step 5. (E)-3-(7-Hydroxy-7-methyl-8-oxo-8,9-dihydro-7H-pyrido[2,3-b]azepin-3-yl)-N-methyl-N ((3-methylbenzofuran-2-yl)methyl)acrylamide (compound 259).

3-bromo-7-hydroxy-7-methyl-7H-pyrido[2,3-b]azepin-8(9H)-one 258 (0.08 g, 0.30 mmol), M-methyl- M-((3-methylbenzofuran-2-yl)methyl)acrylamide 9 (0.07 g, 0.31 mmol), Bu ₄ NCl (9 mg, 0.03 mmol) and Pd-116 (15 mg, 0. 03 mmol), and the flask is evacuated and backfilled with N2 three times. Add 1,4-dioxane (2.5 ml) and DIPEA (0.2 ml, 1.15 mmol) and the reaction mixture is heated to 80°C and stirred for 2 hours. The reaction mixture is cooled to RT, concentrated in vacuo and purified by column chromatography (0-3% MeOH/DCM) to obtain the desired product 259 as a white solid (71 mg, 56%). R _t 1.96 min (method 1a); m/z 418 [M+H]+ (ER+). 1H NMR (400 MHz, DMSO-d, 363 K): δ, ppm 10.52 (s, 1H), 8.60 (d, J=1.9 Hz, 1H), 8.14 (d, J=2.2 Hz, 1H), 7.58-7.50 (m, 2H), 7.46 (d, J=8.0 Hz, 1H), 7.39-7.17 (m, 3H ), 6.72 (d, J=10.9 Hz, 1H), 6.04 (d, J=11.1 Hz, 1H), 5.08 (s, 1H), 4.87 (s, 2H ), 3.11 (s, 3H), 2.27 (s, 3H), 1.22 (s, 3H).

Step 6. Chiral resolution of compound 259.

Chiral separation method: apparatus: Isolera (Biotage). Column: Chiralpak IA (20 µm; glass column; 250 mm x 25 mm). Eluent: CH ₃ CN 90 - Ethyl acetate 10. Flow: 50 ml/min. Temperature: 25°C. Amount injected: 38.4 mg 259 in 10 ml CH3CN 90 - Ethyl acetate 10 and 2 ml MeOH (dissolve at 50°C in ultrasonic bath, then cool and quickly inject).

(R,E-3-(7-Hydroxy-7-methyl-8-oxo-8,9-dihydro-7H-pyrido[2,3-b]azepin-3-yl)-N-methyl-N-( (3-methylbenzofuran-2-yl)methyl)acrylamide (compound 260).

The pure fractions of the first enantiomer are collected and concentrated to dryness. The solid is then suspended in diethyl ether, transferred to a recording tube and concentrated first under a stream of nitrogen, then under vacuum at RT. The first enantiomer is obtained as a white solid (m=13.04 mg; chiral purity: 100%). The absolute configuration is determined arbitrarily.

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Example 55. Synthesis of (E)-3-(3-(hydroxymethyl)-4-oxo-2,3,4,5-tetrahydro-1H-pyrido[2,3b][1,4]diazepin-8-yl) -Nmethyl-N((3-methylbenzofuran-2-yl)megyl)acrylamide (compound 270).

General synthesis scheme.

261 262 263 264

269 270

Reaction conditions: a) TBDPSCI, imidazole, 0°C to RT; b) benzylamine, MeOH, 0°C to RT; c) H ₂ , Pd/C, MeOH, 50°C; d) 5-bromo-3-fluoro-2-nitropyridine 11, TEA, EtOH, reflux; e) Fe, NH ₄ Cl, EtOH, H ₂ O, reflux; f) LiOH, H ₂ O, THF, MeOH; g) GATU, DIPEA, DMF; h) FTAB, THF; i) Pd-116, DIPEA, NBu ₄ Cl, 1,4-dioxane, 80°C.

Step 1. Methyl 2-(((tert-butyldiphenylsilyl)oxy)methyl)acrylate (compound 262).

Imidazole (1.29 g, 19.0 mmol) and tert-butylchlorodiphenylsilane (4.95 mL, 19.0 mmol) were added to a stirred solution of methyl 2-(hydroxymethyl)acrylate 261 (1.77 mL, 17.2 mmol) in DMF (20 ml) at 0°C. The reaction mixture is warmed to RT and stirred for ~16 hours. The reaction mixture is diluted with EtOAc (30 ml) and NaHCO ₃ (30 ml, sat. aq.) is added. The organic phase is separated and the aqueous phase is extracted again with EtOAc (3x30 ml). The combined organic fractions are washed with water (3x50 ml) and brine (2x50 ml), dried using MgSO ₄ and concentrated in vacuo. The crude product was purified by column chromatography (0-50% EtOAc/isohexane) to give the desired product 262 as a colorless oil (5.34 g, 87%). Rt 3.16 min (method 1a) m/z 377 [M+Na]+ (ER+). ^1H NMR (500 MHz, DMSO-de): δ, ppm 7.66-7.59 (m, 4H), 7.52-7.41 (m, 6H), 6.25 (q, J =1.8 Hz, 1H), 6.03 (kv, J=2.0 Hz, 1H), 4.36 (s, 2H), 3.64 (s, 3H), 1.01 (s, 9H ).

Step 2. Methyl 3-(benzylamino)-2-(((tert-butyldiphenylsilyl)oxy)methyl)propanoate (compound 263).

Benzylamine (10% solution in MeOH, 7.4 mL, 6.77 mmol) was added dropwise to a stirred solution of methyl 2-(((tert-butyldiphenylsilyl)oxy)methyl)acrylate 262 (2.4 g, 6.77 mmol ) in MeOH (30 ml) at 0°C, and the reaction mixture was returned to RT. The reaction mixture was stirred for ~16 hours, then concentrated in vacuo, and the crude product was purified by column chromatography (0-20% EtOAc/isohexane) to give the desired compound 263 as a colorless oil (0.99 g, 31%). R _t 2.11 min (method 1a) m/z 462 [M+H]+ (ER+). ^1H NMR (500 MHz, DMSO-dg): δ, ppm 7.62-7.56 (m, 4H), 7.50-7.38 (m, 6H), 7.31-7.17 (m, 5H), 3.88-3.79 (m, 2H), 3.63 (s, 2H), 3.62 (s, 3H), 2.83 (p, J=6.6 Hz, 1H), 2.72 (dd, J=11.7 Hz, 7.0 Hz, 1H), 2.64 (dd, J=11.7 Hz, 6.8 Hz, 1H), 2.17 (s , 1H), 0.94 (s, 9H).

Step 3. Methyl 3-amino-2-(((tert-butyldiphenylsilyl)oxy)methyl)propanoate (compound 264).

A solution of methyl 3-(benzylamino)-2-(((tert-butyldiphenylsilyl)oxy)methyl)propanoate 263 (1.5 g, 3.25 mmol) and palladium on carbon (0.35 g, 3.25 mmol) in MeOH (65 ml) is hydrogenated at 5 bar and 50°C for 24 hours. Palladium is removed by filtration through Celite® and the filtrate is concentrated in vacuo to give the desired product 264 as a mixture of pale yellow oil (1.05 g, 87% ), which is used in the next step without further purification.

Step 4. Methyl 3-((5-bromo-2-nitropyridin-3-yl)amino)-2-(((tert-butyldiphenylsilyl)oxy)methyl)propanoate (compound 265).

To a stirred solution of 5-bromo-3-fluoro-2-nitropyridine 11 (630 mg, 2.83 mmol) in EtOH (15 ml) add methyl 3-amino-2-(((tert-butyldiphenylsilyl)oxy)methyl)propanoate 264 (1.0 g, 2.83 mmol), then TEA (1.0 ml, 7.07 mmol). The reaction mixture is stirred at reflux for 2 hours and then concentrated in vacuo. The crude product was purified by column chromatography (0-100% ethyl acetate/isohexane) to give the title compound 265 as a brown oil (490 mg, 29%). Rt 3.15 min (method 1a) m/z 572/574 [M+H]+ (ER+).

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Step 5. Methyl 3-((2-amino-5-bromopyridin-3-yl)amino)-2-(((tert-butyldiphenylsilyl)oxy)methyl)propanoate (compound 266).

To a mixture of methyl 3-((5-bromo-2-nitropyridin-3-yl)amino)-2-(((tert-butyldiphenylsilyl)oxy)methyl)propanoate 265 (500 mg, 0.85 mmol) add iron (190 mg, 3.42 mmol) and ammonium chloride (185 mg, 3.42 mmol) in ethanol (35 ml) and water (8.8 ml). The resulting reaction mixture is heated at 90°C for 1 hour, then filtered through a pad of Celite® while still hot. The filtrate is then evaporated to dryness. The crude product was purified (0-10% MeOH/DCM) to give the title compound 266 as a colorless solid (300 mg, 64%). R _t 2.59 min (method 1a) m/z 542/544 [M+H]+ (ER+).

Step 6. 3-((2-Amino-5-bromopyridin-3-yl)amino)-2-(((tert-butyldiphenylsilyl)oxy)methyl)propanoic acid (compound 267).

Lithium hydroxide (1 M aq, 618 μL, 0.618 mmol) was added dropwise to the stirred solution of methyl 3-((2-amino-5-bromopyridin-3-yl)amino)-2-(((tert-butyldiphenylsilyl) hydroxy)methyl)propanoate 266 (305 mg, 0.56 mmol) in THF (8.0 ml) and MeOH (1.5 ml). The reaction mixture is stirred at the same temperature for 80 minutes, then stored at -20°C overnight. The mixture is quenched with AcOH (0.5 ml) and the solvents are evaporated in vacuo. The resulting crude product was purified by chromatography (0-10% MeOH/DCM) to give the title compound 267 as a white solid (76 mg, 25%). Rt 1.77 min (method 2a) m/z 528/530 [M+H]+ (ER+).

Step 7. 8-Bromo-3-(((tert-butyldiphenylsilyl)oxy)methyl)-2,3-dihydro-1H-pyrido[2,3-b][1,4]diazepin-4(5H-one ( connection 268).

DIPEA (0.13 mL, 0.72 mmol) was added to a stirred solution of 3-((2-amino-5bromopyridin-3-yl)amino)-2-(((tert-butyldiphenylsilyl)oxy)methyl)propanoic acid 267 ( 76 mg, 0.14 mmol) in DMF (2.0 ml). GATU (66 mg, 0.17 mmol) was added to the mixture and the reaction mixture was stirred for 1 hour. The mixture was quenched with MeOH (4.0 ml) and partitioned between water (65 ml) and EtOAc (50 ml). The aqueous layer was extracted with EtOAc (2x50 ml) and the combined organic extracts were washed with brine (100 ml), dried with MgSO ₄ and evaporated in vacuo to give the title compound 268 as an amber oil (101 mg, 85%), which used in the next step without further purification. R _t 2.33 min (method 2a) m/z 510/512 [M+H]+ (ER+).

Step 8. 8-Bromo-3-(hydroxymethyl)-2,3-dihydro-1H-pyrido[2,3-b][1,4]diazepin-4(5H)-one (compound 269).

Solution of 8-bromo-3 -(((tert-butyldiphenylsilyl)oxy)methyl)-2,3-dihydro-III-pyrido[2,3-b][1,4]diazepin-4(5H-one 268 (101 mg, 0.12 mmol) in THF (4.0 ml) is treated with FTBA (1 M in THF, 0.2 ml, 0.2 mmol). After 20 min, the solvent is evaporated in vacuo and the crude product is purified by chromatography on silica gel ( 0-100% EtOAc/isohexane) to obtain the title compound 269 as a white solid (26 mg, 78%) R _t 0.93 min (method 2a) m/z 272/274 [M+H]+ (ER+).

Step 9. (E)-3-(3-(Hydroxymethyl)-4-oxo-2,3,4,5-tetrahydro-1H-pyrido[2,3-b][1,4]diazepin-8-yl )Nmethyl-N-((3-methylbenzofuran-2-yl)methyl)acrylamide (compound 270).

Namethyl-N-((3-methylbenzofuran-2-yl)methyl)acrylamide 9 (22 mg, 0.096 mmol), 8-bromo-3-(hydroxymethyl)-2,3-dihydro-1H-pyrido[ 2,3-b][1,4]diazepin-4(5H)-one 269 (26 mg, 0.096 mmol), tetrabutylammonium chloride hydrate (2.8 mg, 9.56 μmol), Pd-116 (4.9 mg, 9.56 µmol). The tube is then flushed with nitrogen for 5 minutes. 1,4-Dioxane (2.0 mL) and DIPEA (33 μL, 0.19 mmol) were added and the reaction mixture was heated to 80°C for 1 h, then cooled to RT. The solvent is evaporated in vacuo and the crude product is purified by chromatography (0-10% MeOH/DCM). The resulting product was then triturated from MeCN (2.0 mL) to give the title compound 270 as a pale yellow solid (20 mg, 49%). R _t 1.45 min (method 2a) m/z 421 [M+H]+ (ER+). ^1H NMR (DMSO-de, 363 K): δ, ppm 9.18 (s, 1H), 7.98 (d, J=1.9 Hz, 1H), 7.56 (dd, J= 7.4 Hz, 1.6 Hz, 1H), 7.50-7.39 (m, 2H), 7.38 (d, J=1.9 Hz, 1H), 7.32-7.20 ( m, 2H), 7.12 (d, J=15.4 Hz, 1H), 5.84 (s, 1H), 4.84 (s, 2H), 4.35 (s, 1H), 3, 82-3.74 (m, 1H), 3.60-3.52 (m, 2H), 3.35-3.26 (m, 1H), 3.10 (s, 3H), 2.78- 2.69 (m, 1H), 2.28 (s, 3H).

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Example 56. Synthesis of 2-hydroxyethane-1-aminium ^,E-(2-methyl-8-(3-(methyl((3-methylbenzofuran-2yl)methyl)amino)-3-oxoprop-1-ene-1- yl)-4-oxo-1,2,3,4-tetrahydro-5H-pyrido[2,3-b][1,4]diazepin-5yl)methylphosphate (compound 272).

General synthesis scheme.

Reaction conditions: a) 1. 18-crown-6, NMP/N-ethylpyrrolidone, RT 2. t-BuOK at -20°C to -35°C, then chloromethyl bis-[2-(trimethylsilyl)ethyl]phosphate 21 ; b) TPA/DCM, long-term processing.

Step 1. bis-(2-Trimethylsilylethyl)phosphate [(2R)-2-methyl-8-[(E)-3-[methyl-[(3-methylbenzofuran-2yl)methyl]amino]-3-oxo-prop -1-enyl]-4-oxo-2,3-dihydro-1H-pyrido[2,3-b][1,4]diazepin-5-yl]methyl (compound 271).

Into a 25 ml flask under nitrogen containing (E)-N-methyl-N-[(3-methylbenzofuran-2-yl)methyl]-3-[(2R)-2methyl-4-oxo-1,2,3 ,5-tetrahydropyrido[2,3-b][1,4]diazepin-8-yl]prop-2-enamide 214 (1.0 g, 2.47 mmol, 1 eq.) in suspension in anhydrous DMSO (10 ml) at 25°C, add potassium 2-methylpropan-2-olate solution (2.6 ml, 2.6 mmol, 1.05 eq, 1 M in THF) over 5 min with stirring. The resulting orange solution was then stirred at 25°C for 15 min and added dropwise at 25°C over 30 min to a solution of chloromethylbis(2-(trimethylsilyl)ethyl)phosphate (4.8 mL, 3.46 mmol, 1 .4 eq., 30% in hexane) with vigorous stirring. The reaction mixture is then stirred at 25°C for 10 min until almost complete, as indicated by UPLC/MS (method: 3 min_acid_standard; 254 nm; Rt = 1.56 min (starting material; 7%); Rt = 1, 62 min (not determined; 4%); Rt=1.79 min (not determined; 4%); Rt=2.27 min (other regioisomer; 15%); Rt=2.57 min (expected product; 70% The reaction mixture is washed with n-heptane (2x40 ml), then the DMSO phase is quenched with saturated ammonium acetate (50 ml), and then the aqueous phase is extracted with MTBE (4x50 ml), the combined organic phases are then washed with brine (100 ml). dry over Na ₂ SO ₄ , filter and concentrate on a vacuum rotary evaporator (bath temperature: 20°C) to obtain the crude title compound 271 as a yellow foam (1.686 g, 1.768 mmol, 71.5% yield UPLC). MS (method 1c): 3min_acid_standard; 254 nm;

Step 2. 2-Hydroxyethane-1-aminium ^,E-(2-methyl-8-(3-(methyl((3-methylbenzofuran-2-yl)methyl)amino)-3-oxoprop-1-ene-1 -yl)-4-oxo-1,2,3,4-tetrahydro-5H-pyrido[2,3-b][1,4]diazepin-5-yl)methylphosphate (compound 272).

In a 100 ml three-neck flask under nitrogen containing a solution of bis-(2-trimethylsilylethyl)phosphate [(2R)-2-methyl-8-[(E)-3-[methyl-[(3-methylbenzofuran-2-yl) methyl]amino]-3-oxoprop-1-enyl]-4-oxo-2,3dihydro-1H-pyrido[2,3-b][1,4]diazepin-5-yl]methyl 271 (1686 mg, 1 .77 mmol, 1 eq.) in anhydrous dichloromethane (27.0 ml), at -20°C with stirring, add dropwise a solution of 2,2,2trifluoroacetic acid (2.6 ml, 33.6 mmol, 19 eq.) in anhydrous DCM (27 ml), keeping the temperature below -15°C. The reaction mixture is then stirred at -12°C for 1 hour 30 minutes until complete, as indicated by UPLC/MS (sample prepared in acetonitrile with one drop of ethanolamine; 3 min_acid_standard; 254 nm; Rt=1.35 min (expected product; ER ^- 513.2; 74%); Rt=1.56 min (API; 12%); Rt=1.81 min (semihydrolyzed phosphate)). The temperature was brought to -20°C and a solution of ethanolamine (2.7 ml, 44.22 mmol, 25 eq.) in anhydrous DCM (27 ml) was added over 5 minutes. The reaction mixture is then stirred at -15°C for 1 hour and then concentrated (rotary evaporator) under high vacuum while maintaining the temp. baths 5°C. The residue was then suspended in 100 ml of cyclopentyl methyl ether and the resulting suspension was filtered to obtain the TPA salt of ethanolamine as a yellowish solid. The filtrate is concentrated (rotary evaporator) under high vacuum, with

- 127 046887 storing temp. 5°C baths to give the crude title product as a yellow oil m=3.76 g (UPLC/MS; 3 min_acid_standard; 254 nm; purity: 87%). Store at -20°C overnight. The product is purified by flash chromatography using Isolera equipment (Biotage). Cartridge: C18 (25 µm); 120 g. Injection: The crude product (3.76 g) is solubilized in 25 ml (water 0.2% ethanolamine - C.'H ₃ C.'N 0.2% ethanolamine (95-5)) before injection. Fractions of interest were combined and lyophilized to give the title product 272 as a yellow solid (m=738 mg), which showed excess ethanolamine by NMR. The product is suspended in methanol (5 ml), sonicated and the suspension filtered. The resulting pale yellow powder was washed with methanol (2x5 ml) and dried under vacuum to obtain the title compound 272 as a pale yellow powder (m=300.3 mg; 26.7% yield). UPLC-MS (method 1c): 3 min_acid_standard; 254 nm; purity 100% (PPC), ER ^- : 513.2. ^1H NMR (400 MHz, D2O): δ, ppm, 8.26 (1H, dd, J=1.8, 15.0 Hz), 7.63 (1H, dd, J=2.0, 16.2 Hz), 7.54-7.32 (3H, m), 7.29-7.16 (2.5H, m), 7.04 (0.5H, d, J=15.6 Hz ), 5.48-5.37 (2H, m), 4.81 (0.8H, s), 4.73 (1.2H, s), 3.99 (1H, kvd, J=6.1 , 18.2 Hz), 3.72 (4H, t, J=5.4 Hz), 3.15 (1.6H, s), 3.04 (4H, t, J=5.0 Hz), 3.00 (1.4H, s), 2.64-2.55 (1H, m), 2.31-2.22 (1H, m), 2.19 (1.6H, s), 2, 18 (1.4H, s), 1.20 (3H, dd, J=2.8, 6.1 Hz). Content of TPA salt of ethanolamine: 0.07 w/w%.

Example 57. Synthesis of (E)-N-[(4-amino-3-methyl-benzofuran-2-yl)methyl]-3-[(3S)-3-hydroxy-4-oxo^DH-tetrahydropyridoCD-CV^ diazepin^-ylf^methylprop^-enamide (compound 274).

274 275

Reaction conditions: a) Pd-162, DIPEA, n-Bu ₄ NCl, 1,4-dioxane, 90°C, 5 h; TPA/DXM; c) DIPEA, GATU, DMF, ct.

Step 1. tert-Butyl (S,E)-3-(3-hydroxy-4-oxo-2,3,4,5-tetrahydro-1H-pyrido[2,3-b][1,4]diazepine8- yl)acrylate (compound 273).

Mixture of (3S)-8-bromo-3-hydroxy-1,2,3,5-tetrahydropyrido[2,3-b][1,4]diazepin-4-one 228 (182.76 mg, 0.70 mmol , 1 eq.), tetrabutylammonium chloride (23.39 mg, 0.08 mmol, 0.12 eq.), chloro(crotyl)(tri-tert-butylphosphine) palladium(P) (14.72 mg, 0.04 mmol, 0.05 eq.) is added to the flask under nitrogen atmosphere. The flask is purged and backfilled with nitrogen. Add 1,4-dioxane (4.95 ml), tert-butyl acrylate 15 (0.24 ml, 1.68 mmol, 2.4 eq) and DIPEA (0.24 ml, 1.4 mmol, 2. 0 eq.), and nitrogen was bubbled through the mixture for 5 minutes. The reaction mixture is heated at 90°C for 5 hours. After cooling, the mixture is filtered using 0.22 μM PTFE filters. The filters are then washed with DCM/MeOH 50/50. The combined filtrate is concentrated under reduced pressure to dryness. ACN is added and the resulting yellow suspension is placed in an ultrasonic bath for a few minutes and then filtered through a porosity 3 frit. The resulting yellow solid is washed with cold ether to give 273 (110 mg, 0.35 mmol, 51.07% yield) as yellow solid. UPLC-MS (method 1c): m/z 306.1 [M+H]+ (ER+).

Stage 2. (S,E)-3-(3-Hydroxy-4-oxo-2,3,4,5-tetrahydro-1H-pyrido[2,3-b][1,4]diazepin-8yl)acrylic acid (compound 274).

tert-Butyl (S,E)-3-(3-hydroxy-4-oxo-2,3,4,5-tetrahydro-1H-pyrido[2,3-b][1,4]diazepin-8yl)acrylate 273 (110 mg, 0.36 mmol, 1.0 eq.) was diluted with DCM (0.23 ml) and cooled to 0°C. Trifluoroacetic acid (0.23 ml) is added dropwise. The resulting yellow solution was stirred for 3 hours. UPLC analysis showed complete conversion. The volatiles are removed under reduced pressure and the resulting residue is treated with cold ether. The resulting suspension is placed in an ultrasonic bath and filtered. The resulting yellow solid was dried overnight under high vacuum to give the title compound 274 (90 mg, 0.35 mmol, 98.23% yield) as a yellow solid. UPLC-MS (method 1c): m/z 250.1 [M+H]+ (ER+).

Step 3. (E)-N-[(4-Amino-3-methyl-benzofuran-2-yl)methyl]-3-[(3S)-3-hydroxy-4-oxo-1,2,3,5tetrahydropyridoCD -CPDCiazepin^-yl^^methyl-prop^-enamide (compound 275), 3-methyl-2(methylaminomethyl)benzofuran-4-amine 3 (32.1 mg, 0.169 mmol, 1.2 eq.) and (^ ,E)-3-(3-hydroxy-4-oxo2,3,4,5-tetrahydro-1H-pyrido[2,3-b][1,4]diazepin-8-yl)acrylic acid 274 (35, 1 mg, 0.141 mmol, 1.0 eq.) dissolved in DMF (0.7 ml). GATU (83.76 mg, 0.21 mmol, 1.5 eq.) is then added in one portion. DIPEA (0.098 ml, 0.564 mmol, 4.0 eq.) was added dropwise at RT, and the resulting mixture was stirred at RT for 1 hour. The resulting brown reaction mass was concentrated in

- 128 046887 high vacuum (no treatment), the crude brown product was dissolved in MeCN/H2O (5 ml, 4:1 v/v, 2 injections) and purified by semipreparative HPLC (98-100-30 min; 90% solvent A, 10% solvent B, 30 min) and lyophilized overnight to give the title compound 275 as a yellow powder with a purity of 89.19% (49 mg, 82% yield). The resulting solid is dried and solubilized in DMSO (0.5-1 ml) and slowly added to water (6 ml). The rapidly precipitated product with a purity of 93.14% was recovered by centrifugation and lyophilized overnight. A beige powder (10.5 mg) is obtained. UPLC-MS (method 1c): m/z 422.1 [M+H]+ (ER ⁺ ); tret=1.02 min. ¹ H NMR (400 MHz, DMSO-ph): δ, ppm 10.07 (s, 1H), 7.97-7.94 (m, 1H), 7.37-7.28 (m, 2 ,4H), 7.06 (d, J=15.4 Hz, 0.6H), 6.84 (t, J=8.0 Hz, 1H), 6.58-6.56 (d, 1H) , 6.30 (d, J=7.8 Hz, 1H), 6.02 (shm, 1H), 5.13-5.11 (m, 3H), 4.77, 4.62 (rotamers, 2c , 2H), 4.08-4.06 (m, 1H), 3.41-3.38 (m, 1H), 3.21-3.16 (m, 1H), 3.01, 2.84 (rotamers, 2a, 3H), 2.34 (s, 3H).

Example 58. Synthesis of N,E)-N-((7-amino-2-methylbenzofuran-3-yl)methyl)-3-(3-hydroxy-4-oxo-2,3,4,5tetrahydro-1H-pyrido [2,3-b][1,4]diazepin-8-yl)-N-methylacrylamide (compound 276).

Reaction conditions: a) DIPEA, GATU, DMF, rt.

Step 1. (S, E)-N-((7-Amino-2-methylbenzofuran-3-yl)methyl)-3-(3-hydroxy-4-oxo-2,3,4,5tetrahydro-1H-pyrido [2,3-b][1,4]diazepin-8-yl)-N-methylacrylamide (compound 276).

2-Methyl-3-((methylamino)methyl)benzofuran-7-amine 74 (31 mg, 0.164 mmol, 1.2 eq.) and (S,E)-3-(3Hydroxy-4-oxo-2,3 ,4,5-tetrahydro-1H-pyrido[2,3-b][1,4]diazepin-8-yl)acrylic acid 274 (34.14 mg, 0.137 mmol, 1.0 eq.) dissolved in DMF ( 0.7 ml). GATU (81.37 mg, 1.5 mmol, 1.5 eq.) is then added in one portion. There, DIPEA (0.096 ml, 0.54 mmol, 4.0 eq.) was added dropwise at RT, and the resulting mixture was stirred at RT for 2 hours. The resulting brown reaction mass was treated with water (5 ml). The rapidly precipitated product is collected by centrifugation. A yellow solid was isolated with a purity of 66.53% (48 mg). The resulting product with impurities is re-purified by preparative HPLC. The yellow product is suspended in 4 ml MeOH/DCM (sonication 1-2 min) and transferred to a 4 ml barcoded tube. After evaporation of the solvent and drying under high vacuum overnight at RT, a yellow solid was obtained with 100% purity (9.6 mg, 16.6% yield). 1H NMR (400 MHz, DMSO-ph): δ, ppm 10.14 (s, 1H), 8.05-8.01 (m, 1H), 7.49-7.38 (m, 2.3H ), 7.06 (d, J=15.4 Hz, 0.7H), 6.84 (t, J=7.7 Hz, 1H), 6.74, 6.57 (2 d, 1H), 6.48 (d, J=7.5 Hz, 1H), 6.08 (shm, 1H), 5.20 (d, J=4.6 Hz, 2H), 5.16 (s, 1H), 4.82, 4.67 (rotamers, 2c, 2H), 4.15-4-12 (m, 1H), 3.47 (ddd, J=3.3, 6.1 and 12.2 Hz, 1H ), 3.28-3.22 (m, 1H), 3.01, 2.82 (rotamers, 2c, 3H), 2.50, 2.45 (rotamers, 2c, 3H). UPLCMS (method 1c): m/z 422.1 [M+H] ⁺ (ER ⁺ ); t _ret =1.00 min.

Example 59. Synthesis of decanoate ^,E-(3-hydroxy-3-methyl-8-(3-(methyl((3-methylbenzofuran-2-yl)methyl)amino)-3-oxoprop-1-en-1- yl)-4-oxo-1,2,3,4-tetrahydro-5H-pyrido[2,3-b][1,4]diazepin-5-yl)methyl (compound 282).

Reaction conditions: a) n-Bu ₄ N-HSO ₄ , NaHCO ₃ , DCM/H ₂ O (1:1 v/v), 0°C to RT, overnight; b) 1-methyl-III-imidazole, DMSO, QD; c) tBuOK, DMSO, RT, 30 min; d) 1M aq. HCl.

Step 1. Chloromethyldecanoate (compound 279). Decanoic acid 277 (1.0 g, 5.69 mmol, 1.0 eq.), tetrabutylammonium hydrogen sulfate (99.57 mg, 0.28 mmol, 0.05 eq.) and sodium bicarbonate (1.92 g,

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22.76 mmol, 4.0 eq.) are mixed in a flask. Add DCM (20 ml) and water (20 ml). The resulting two-phase solution was cooled to 0°C and then treated with a solution of chloromethyl sulfur chloridate 278 (1.03 g, 6.26 mmol, 1.1 eq.) in DCM (0.5 ml). The mixture is stirred at 0°C for 40 minutes and then at RT overnight. The reaction mixture is diluted with water and extracted twice with DCM. The combined organic layers were dried over MgSO ₄ to give chloromethyl decanoate 279 (0.89 g, 4.06 mmol, 71.35% yield) as a colorless oil.

Stage 2. Sh,E-H-Methyl-3-(3-methyl-4-oxo-3-((trimethylsilyl)oxy)-2,3,4,5-tetrahydro-Spirido[2,3-b][ 1,4]diazepin-8-yl)-N-((3-methylbenzofuran-2-yl)methyl)acrylamide (compound 280).

To 1-methyl-III-imidazole (1.33 mL, 16.65 mmol, 7.0 eq.) in solution in anhydrous DMSO (10 mL) add (E)-3-[(3R)-3-hydroxy- 3-methyl-4-oxo-2,5-dihydro-1H-pyrido[2,3-b][1,4]diazepin-8-yl]-Nmethyl-N-[(3-methylbenzofuran-2-yl) methyl]prop-2-enamide 49 (1.0 g, 2.38 mmol, 1.0 eq.) in one serving at RT. The yellow solution was stirred at RT for 5 min, then TMSCl (1.54 mL, 11.89 mmol, 5.0 eq.) was added in one portion. The reaction mixture was stirred at RT for 1 hour, diluted with water and brine (1:1 v/v) and extracted with AcOEt (3x). The combined organic phases were washed with brine, dried over MgSO4, filtered and concentrated to dryness to give the title compound 280 (1.12 g, 2.27 mmol, 95.59% yield) as a yellow solid. UPLC-MS (method 1c): R _t =2.04 min. ER ⁺ = 493.3.

Step 3. (R,E-(3-Methyl-8-(3-(methyl((3-methylbenzofuran-2-yl)methyl)amino)-3-oxoprop-1-en-1 yl)-4-oxo -3 -((trimethylsilyl)oxy)-1,2,3,4-tetrahydro-5H-pyrido[2,3-b][1,4]diazepin-5-yl)methyldecanoate (compound 281).

(R,E-N-Methyl-3-(3-methyl-4-oxo-3-((mrimethylsilyl)oxy)-2,3,4,5-tetrahydro-1H-pyrido[2,3-b][ 1,4]diazepin-8-yl)-H-((3-methylbenzofuran-2-yl)methyl)acrylamide 280 (0.569 g, 1.16 mmol, 1.0 eq.) was diluted with DMSO (5 ml B). the resulting yellow solution was added with potassium 2-methylpropan-2-olate (150.07 mg, 1.27 mmol, 1.1 eq.) This reaction mixture was added dropwise over 7 min to the test tube containing chloromethyl decanoate 279 (1.02). g, 4.62 mmol, 4.0 eq.) The reaction mixture was stirred at RT for 30 min, diluted with water and extracted twice with EtOAc. The combined organic layers were washed with brine, dried over MgSO ₄ and the residue was concentrated to dryness. silica gel using a SNAP Ultra column (50 g) and DCM/EtOAc 90/10-0/100 as eluent. Good fractions were collected and concentrated to dryness to give product 281 as a beige foam (67 mg, 24% yield). MS (method 1c): R _t =2.91 min ER ⁺ = 677.5.

Step 4. (R,E-(3 -G hydroxy-3 -methyl-8-(3 -(methyl((3 -methylbenzofuran-2-yl)methyl)amino)-3 oxoprop-1-en-1-yl )-4-oxo-1,2,3,4-tetrahydro-5H-pyrido[2,3-b][1,4]diazepin-5-yl)methyldecanoate (compound 282).

(R,E-(3 -Methyl-8-(3 -(methyl((3-methylbenzofuran-2-yl)methyl)amino)-3 -oxoprop-1 -en-1-yl)-4-oxo-3 ((trimethylsilyl)oxy)-1,2,3,4-tetrahydro-5H-pyrido[2,3-b][1,4]diazepin-5-yl)methyldecanoate 281 was dissolved in MeOH (2 ml). HCl in water (0.58 mL, 0.57 mmol, 2.0 eq.) was added at RT. After 10 min of stirring at room temperature, UPLC analysis indicated complete conversion. The reaction mixture was concentrated to dryness to give a yellow oil, which was diluted with EtOAc. and washed twice with saturated aq. NaHCO ₃ solution. The combined organic layers are dried over MgSO ₄ and concentrated to dryness. This crude product is purified by flash chromatography using a SNAP Ultra column (25 g) and DCM/AcOEt 50/50-0/100 in. as eluent, the good fractions were collected and concentrated to dryness to give lipophilic prodrug 282 as a beige solid (103 mg, 0.16 mmol, 56.17% yield UPLC-MS (method 1c): R _t =2.46 min). ER ⁺ = 605.4 ^1H NMR (400 MHz, DMSO-de): δ, ppm, 7.88-7.85 (m, 1H), 7.65-7.63 (m, 1H) , 7.60 (d, J=15.6 Hz, 1H), 7.50 (d, J=7.3 Hz, 1H), 7.40 (d, J=7.9 Hz, 1H), 7 .28-7.20 (m, 2H), 7.20-7.06 (m, 1H), 6.79 (d, J=15.4 Hz, 1H), 5.26-5.17 (m , 2H), 4.86, 4.69 (2c, 2H, rotamers), 4.08 (shs, 1H), 3.72 (d, J=11.9 Hz, 1H), 3.25 (d, J=12.4 Hz, 1H), 3.21, 3.07 (2s, 3H, rotamers), 2.30 (s, 3H), 2.23-2.19 (m, 2H), 1.61 (s, 3H), 1.55-1.51 (m, 2H), 1.15-1.35 (m, 12 H), 0.86 (t, J=6.6 Hz, 3H).

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Example 60. Synthesis of (K,E-(3-g'hydroxy-3-methyl-8-(3-(methyl((3-methylbenzofuran-2-yl)methyl)amino)2-ethylbu3-oxoprop-1-ene -1-yl)-4-oxo-1,2,3,4-tetrahydro-5H-pyrido[2,3-b][1,4]diazepin-5-yl)methyl tanoate (compound 286).

General synthesis scheme.

Reaction conditions: a) n-Bu4N-HSO ₄ , NaHCO ₃ , DCM/H ₂ O (1:1 v/v), 0°C to RT, overnight; b) tBuOK, DMSO, RT, 30 min; c) 1 M aq. HCl.

Step 1: Chloromethyl 2-ethylbutanoate (compound 284) is synthesized from 2-ethylbutanoic acid (283) and chloromethyl sulfur chloridate 278 as described for compound 279 (77.5% yield).

Step 2.(R, E)-(3-Methyl-8-(3-(methyl((3-methylbenzofuran-2-yl)methyl)amino)-3-oxoprop-1-en-1 yl)-4- Oxo-3-((trimethylsilyl)oxy)-1,2,3,4-tetrahydro-5H-pyrido[2,3-b][1,4]diazepin-5-yl)methyl 2ethylbutanoate (compound 285) was prepared according to the procedure described for compound 281 (49% yield). UPLC-MS (method 1c): R _t =2.54 min ER ⁺ = 621.4.

Step 3. ^,E-(3-Hydroxy-3-methyl-8-(3-(methyl((3-methylbenzofuran-2-yl)methyl)amino)-3oxoprop-1-en-1-yl)-4 -Oxo-1,2,3,4-tetrahydro-5H-pyrido[2,3-b][1,4]diazepin-5-yl)methyl 2-ethylbutanoate (compound 286) was prepared as described for compound 282 ( 79% yield).

UPLC-MS (method 1c): Rt=1.96 min. ER ⁺ = 549.3. 1H NMR (400 MHz, DMSO-dQ: δ, ppm 7.87-7.84 (m, 1H), 7.67 (t, J=6.9 Hz, 1H), 7.59 (d, J=15.4 Hz, 1H), 7.48 (d, J=7.2 Hz, 1H), 7.39 (d, J=7.9 Hz, 1H), 7.287.20 (m, 2H) , 7.06-7.04 (m, 1.3H), 6.79 (d, J=15.4 Hz, 0.7H), 5.27-5.19 (m, 2H), 4.82 , 4.69 (2c, 2H, rotamers), 4.08 (shs, 1H), 3.76 (d, J=11.9 Hz, 1H), 3.21-3.18 (m, 1H), 3.20, 3.08 (2s, 3H, rotamers), 2.30 (s, 3H), 2.152.09 (m, 1H), 1.59 (s, 3H), 1.56-1.37 ( m, 4H), 0.81-0.72 (m, 6H).

Example 61. Synthesis of (E)-N-((7-amino-2-methylbenzofuran-3-yl)methyl)-3-(2,2-dimethyl-3-oxo-1,2,3,4tetrahydropyrido[2, 3-b]pyrazin-7-yl)-N-methylacrylamide (compound 287).

General synthesis scheme.

166 14Reaction conditions: a) i) Pd-116, DIPEA, 1,4-dioxane, 80°C; ii) HCl, MeOH.

Step 1. (E)-N-((7-Amino-2-methylbenzofuran-3-yl)methyl)-3 -(2,2-dimethyl-3-oxo-1,2,3,4tetrahydropyrido[2,3 -b]pyrazin-7-yl)-N-methylacrylamide (compound 287).

To a mixture of N((7-((diphenylmethylene)amino)-2-methylbenzofuran-3-yl)methyl)-X-methylacrylamide 166 (72 mg, 0.18 mmol), 7-bromo-2,2-dimethyl-1 ,2-dihydropyrido[2,3-b]pyrazin-3(4H)-one 14 (45 mg, 0.18 mmol) and Pd-116 (9 mg, 0.02 mmol) add 1,4-dioxane (2 ml) and DIPEA (0.06 ml, 0.35 mmol). The reaction mixture is purged with N ₂ , heated to 80°C and stirred for 1 hour. The reaction mixture is cooled to RT, then diluted with MeOH (2 ml) and HCl added (5 ml, 1 M aq.). The resulting mixture was stirred for 10 min, then diluted with HCl (5 ml, 1 M aq.). Aqueous extract layer

- 131 046887 add DCM (3x10 ml), neutralize with solid NaHCO ₃ to ~pH 8 and extract with DCM (3x10 ml). The combined organic layers were washed with brine (20 mL), passed through a phase separator and concentrated in vacuo. The crude product was purified by chromatography (0-10% MeOH/DCM) to give the title compound 287 as a pale yellow solid (26 mg, 34%). R _t 1.51 min (method 1a) m/z 420 (M+H)+ (ER ⁺ ). ^1H NMR (DMSOD ₆ , 363 K): δ, ppm 10.41 (s, 1H), 7.88 (d, J=1.9 Hz, 1H), 7.46 (d, J=15 .4 Hz, 1H), 7.24 (d, J=1.9 Hz, 1H), 7.03 (d, J=15.4 Hz, 1H), 6.88 (t, J=7.7 Hz, 1H), 6.75 (d, J=7.7 Hz, 1H), 6.54 (dd, J=7.8, 1.2 Hz, 1H), 6.04 (s, 1H), 4.88 (s, 2H), 4.72 (s, 2H), 2.98 (s, 3H), 2.47 (s, 3H), 1.30 (s, 6H).

Example 62. Synthesis of (E)-N-((7-amino-2-methylbenzofuran-3-yl)methyl)-3-((2R,3S)-3-hydroxy-2-methyl4-oxo-2,3, 4,5-tetrahydro-1H-pyrido[2,3-b][1,4]diazepin-8-yl)-N-methylacrylamide (compound 290).

General synthesis scheme.

148 288

289 290

Reaction conditions: a) Pd-116, DIPEA, 1,4-dioxane, 90°C; b) TPA, DXM; c) GATU, DIPEA, DMF.

Step 1. N-tert-Butyl 3-((2R,3S)-3-hydroxy-2-methyl-4-oxo-2,3,4,5-tetrahydro-1H-pyrido[2,3b][1, 4]diazepin-8-yl)acrylate (compound 288).

(2R,3 S)-8-BpoM-3-hydroxy-2-methyl-2,3-dihydro-1H-pyrido[2,3-b][1,4]diazepin-4(5H)-one 148 (350 mg, 1.29 mmol) and Pd-116 (33 mg, 0.064 mmol) are added to the tube and the tube is evacuated and refilled with nitrogen (3 times). 1,4-Dioxane (7 ml), DIPEA (0.45 ml, 2.57 mmol) and tert-butyl acrylate 15 (0.37 ml, 2.57 mmol) were added and the reaction mixture was stirred at 90°C for 1 h The reaction mixture is cooled to RT and then concentrated in vacuo. The crude product was purified by chromatography (0-50% EtOAc/isohexane) to give the title compound 288 as a yellow solid (414 mg, 96%). UPLC-MS (method 1c): Rt=1.74 min. m/z 320 [M+H] ⁺ (ER ⁺ ). ¹ H NMR (500 MHz, DMSO-06): δ, ppm 10.30 (s, 1H), 7.97 (d, J=1.9 Hz, 1H), 7.43 (d, J= 16.0 Hz, 1H), 7.36 (d, J=1.9 Hz, 1H), 6.35 (d, J=16.0 Hz, 1H), 6.11 (d, J=5, 7 Hz, 1H), 5.15 (d, J=4.8 Hz, 1H), 4.18 (dd, J=4.8, 3.3 Hz, 1H), 3.77-3.69 ( m, 1H), 1.48 (s, 9H), 1.08 (d, J=6.5 Hz, 3H).

Step 2. 2,2,2-Trifluoroacetate (E)-3-((2R,3S)-3-hydroxy-2-methyl-4-oxo-2,3,4,5-tetrahydro-1Hpyrido[2,3 -b][1,4]diazepin-8-yl)acrylic acid (compound 289).

A mixture of N-tert-butyl 3-((2R,3S)-3-hydroxy-2-methyl-4-oxo-2,3,4,5-tetrahydro-1H-pyrido[2,3b][1,4] diazepin-8-yl)acrylate 288 (411 mg, 1.29 mmol) in DCM (5 ml) and TFA (5 ml) stir at RT for 1.5 hours. The mixture is concentrated under reduced pressure to give a pale orange gum . The gum is triturated with TBME (10 ml) to obtain a yellow solid, which is filtered and washed with more TBME (10 ml) and then MeCN (2x10 ml) to obtain the title compound 289 as a bright yellow solid (260 mg, 53 %). UPLC-MS (method 1c): R _t =0.90 min. m/z 264 [M+H] ⁺ (ER ⁺ ). 1H NMR (500 MHz, DMSO-06): δ, ppm 12.41 (s, 1H), 10.30 (s, 1H), 7.96 (d, J=2.0 Hz, 1H), 7.47 (d, J=16.0 Hz, 1H), 7.36 (d, J=2.0 Hz, 1H), 6.36 (d, J=16.0 Hz, 1H), 6, 15 (d, J=5.7 Hz, 1H), 5.15 (shs, 1H), 4.19 (d, J=3.3 Hz, 1H), 3.78-3.69 (m, 1H ), 1.08 (d, J=6.5 Hz, 3H).

Step 3. (E)-N-((7-Amino-2-methylbenzofuran-3-yl)methyl)-3-((2R,3S)-3-hydroxy-2-methyl-4-oxo2,3,4 ,5-tetrahydro-1H-pyrido[2,3-b][1,4]diazepin-8-yl)-N-methylacrylamide (compound 290).

DIPEA (110 µl, 0.63 mmol) was added to the suspension of 2,2,2-trifluoroacetate (E)-3-((2R,3S)-3hydroxy-2-methyl-4-oxo-2,3,4,5 -tetrahydro-1H-pyrido[2,3-b][1,4]diazepin-8-yl)acrylic acid 289 (60 mg, 0.16 mmol) and GATU (72 mg, 0.19 mmol) in DMF ( 1 ml), and the mixture was stirred at RT for 2 minutes. 2-Methyl-3-((methylamino)methyl)benzofuran-7-amine 74 (30 mg, 0.16 mmol) in DMF (1 ml) was added in one portion and the reaction mixture was stirred at RT for 1.5 hours. H2O (10 ml) is added and the resulting yellow solid is filtered and washed with H2O (5 ml). The crude product was purified by chromatography (2.5-5% MeOH/DCM) to give the title compound 290 as a yellow solid (16 mg, 23%). R _t 1.36 min (method 1a) m/z 436 (M+H)+ (ER ⁺ ). 1H NMR (500 MHz, DMSO, 363 K): δ, ppm 9.80 (s, 1H), 7.97 (d, J=1.9 Hz, 1H), 7.44 (d, J= 15.4 Hz, 1H), 7.39 (d, J=1.9 Hz, 1H), 7.15-7.03 (m, 1H), 6.87 (t, J=7.7 Hz, 1H), 6.74 (d, J=7.7 Hz, 1H), 6.53 (d, J=7.7 Hz, 1H), 5.89 (d, J=5.7 Hz, 1H) , 4.87 (s, 2H), 4.76 (d, J=4.8 Hz, 1H), 4.71 (s, 2H), 4.21 (t, J=3.9 Hz, 1H) , 3.80-3.72 (m, 1H), 2.98 (s, 3H), 2.47 (s, 3H), 1.11 (d, J=6.5 Hz, 3H).

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Example 63 Examples of compositions.

The compound of the present invention is formulated as a solution at a concentration of 10 mg/ml in 40% Captisol aqueous vehicle or 30% Kleptose vehicle (30 minutes of magnetic stirring at room temperature). This composition is suitable for IV administration.

The compound of the present invention is formulated as a solid dispersion at 20% drug load in HPMC AS: the compound and polymer are solubilized in methylene chloride:methanol (ratio 3:1 w/w), the polymer concentration is 1.4 w/w. % in organic phase (batch scale 85 mg). This solution is then spray dried on a Procept 4M8Trix spray dryer (process parameters: cyclone S, air speed 0.35 m ³ /min, inlet temperature 50°C, feed solution speed: 6 g/min, nozzle pressure 10 l/min, air cooling 80 l/min). The recovery is 64%. DSC and PRD analyzes confirm the receipt of a solution of a solid (amorphous physical form, single Tg at 95°C). The composition is then formulated into tablets or granules for PO administration.

The compound of the present invention is formulated (at a concentration of 1 mg/ml to 300 mg/ml) as a nanosuspension or microsuspension in water or oil and stabilized with a polymer such as cellulose, 2-hydroxypropyl ether or cellulose ethyl ether at a concentration of 0.01 to 10%. This composition additionally contains a surfactant such as polyoxyethylene 20 sorbitan monooleate in a concentration of 0.01 and 10%. This formulation is suitable for oral administration.

The compound of the present invention is formulated (at a concentration of 1 mg/ml to 300 mg/ml) as a nanosuspension or microsuspension in water or oil. The composition further contains a polymer such as polyethylene glycol 4000 or a-hydro-o-hydroxypoly(oxy-1,2-ethanediyl)cellulose at a concentration of 0.01 to 10%, and a surfactant such as polyoxyethylene 20 sorbitan monooleate concentrations from 0.01 and 10%. The formulation is a slow release formulation (slow release 12 to 72 hours). This composition is suitable for intramuscular administration.

The compound of the present invention is formulated as a nanosuspension or microemulsion in water at a concentration of 10 mg/ml. The composition additionally contains polyethylene glycol 4000 at a concentration of 10% and Polyoxyethylene 20 sorbitan monooleate at a concentration of 10%. This composition is suitable for intramuscular administration.

The compound of the present invention is formulated (at a concentration of 1 mg/ml to 300 mg/ml) as a nanosuspension or microemulsion in water in combination with a hydrophilic prodrug of the present invention in a (compound:hydrophilic prodrug) ratio of 1:99 to 99:1 . The composition further contains a polymer, such as copovidone or a polymer of ethenyl ester of acetic acid with 1-ethenyl-2-pyrrolidinone in a concentration of from 0.01 to 10%, and a surfactant, for example Polyoxyethylene 20 sorbitan monooleate, in a concentration from 0.01 to 10%. The formulation is a slow release formulation (slow release 12 to 72 hours). This composition is suitable for intramuscular administration.

The compound of the present invention is formulated (at a concentration of 1 mg/ml to 300 mg/ml) as a solid dispersion made from granules, microspheres or implant, at a drug load of 10-95% within PLGA (or other suitable matrix such like PLA) in an aqueous or oily medium such as hemp oil. The formulation is stabilized with a polymer such as copovidone or a polymer of ethenyl ester of acetic acid with 1-ethenyl-2-pyrrolidinone, in a concentration of 0.01 and 10%. This composition additionally contains the surfactant Polyoxyethylene 20 sorbitan monooleate in concentrations of 0.01 and 10%. The formulation is a sustained release formulation (delayed release from 12 to 72 hours, for example 24 and 72 hours), this formulation is suitable for administration by the intramuscular route.

The compound of the present invention is formulated (at a concentration of 1 mg/ml to 300 mg/ml) as a hydrogel or oleogel containing a polymer such as PEG-PLA polyethylene glycol polylactic acid, or polysaccharides at a concentration of 0.01 to 50%. The composition is a slow release formulation (slow release of 12 to 72 hours, for example 24 and 72 hours). This composition is suitable for intramuscular administration.

The compound of the present invention is formulated as a lipophilic solid or an oily prodrug (at a concentration of 1 mg/ml to 300 mg/ml) with an oil such as hemp oil and a cosolvent such as benzyl alcohol and/or ethanol. The formulation is a slow release formulation (slow release 12 to 72 hours). This composition is suitable for intramuscular administration.

Example 64 Antibacterial activity.

Typical compounds are tested for activity based on the following test procedures.

Neisseria gonorrhoeae FabI inhibition assay.

FabI inhibition of Neisseria gonorrhoeae enzyme activity is tested by measuring the rate of NADH consumption (Δ absorbance at 340 nm/min) at 30°C in a 96-well plate using an automated plate reader in the presence or absence of test compounds.

- 133 046887

The assay mixture contains 100 mM Tris-HCl, pH 7.2, 100 mM ammonium acetate, 0.05% pluronic F-68, 25 μM crotonyl ACP, 50 μM NADH, 50 pM recombinant FabI protein N. gonorrhoeae, 7.5 % DMSO and test compounds at concentrations ranging from 0.17 to 10,000 nM in a final well volume of 100 µl. This dose-dependent inhibition assay is performed using an 11-point serial dilution series for each test compound. IC ₅₀ values for each test compound are determined from a logistic sigmoid curve fit from the dose-dependent inhibition curves.

MIC50. MIC is determined according to appropriate Clinical and Laboratory Standards Institute (CLSI) methodology through agar dilution sensitivity testing method in GC agar base. The isolate screening panel, used to evaluate activity against different strains of Neisseria gonorrhoeae and to determine MIC50, contains a minimum of 15 clinical isolates of N. gonorrhoea, including several strains from the ATCC reference collection and a strain resistant to Ciprofloxacin. The MIC50 is the MIC value at which >50% of the isolates in the test population are inhibited; it is equivalent to the median MIC value.

Caco-2 assay. The compounds are incubated in the 24-transvel Caco-2 system (Solvo Biotechnologies, Hungary) for 120 min, after which samples are taken from the donor and acceptor compartments of the transvel system. The integrity of each cell monolayer is monitored through transepithelial electrical resistance (TEER) measurements prior to the experiment. The permeability of Lucifer yellow (B to A) is also tested as a control on the same plate (all wells) after the actual experiment. Samples are analyzed using UPLC/MS/MS. The permeability of compounds is monitored using relative LC/MS/MS peak areas (donor stock sample=100%). The data is used to calculate apparent permeability (Papp in cm/s x 10 ^-6 ) and recovery during the experiment.

Prediction of liver extraction ratio (ER). In vitro metabolic stability is assessed by determining the intrinsic metabolic clearance after incubation of test compounds with mouse and human liver microsomes. Briefly, test compounds are incubated with liver microsomes supplemented with cofactors (1 mM NADPH, 1 mM UDPGA + 15 mM alamethicin). At selected time points, assays were stopped using an equal volume of cold acetonitrile and samples were analyzed by UPLC/HR-MS to monitor the rate of disappearance of compounds. Substrate depletion is estimated based on peak areas (0 min=100%) and is used to calculate half-lives and then in vitro clearances (CLint, in µl/min/mg protein). Using species-specific physiologically based scaling factors (PBSF) and the Well-Stirred hepatic clearance model modified by Poulin et al, Journal of Pharmaceutical Sciences, vol. 101, no. 2, February 2012 and Poulin et al., Journal of Pharmaceutical Sciences, vol. 101, no. 11, November 2012, in vitro CLint is then transformed into in vivo CLint and liver extraction ratio (ER) for mouse and human.

Determination of PK and oral bioavailability on a mouse cassette: Female BALB/c mice (n=24 per group) receive one intravenous (IV, 1 mg/kg) or oral (2 mg/kg) dose of compounds. Dose volumes are 5 ml/kg and 10 ml/kg for IV and oral administration, respectively. Compounds are co-formulated in 10% DMSO (v/v) and 40% Captisol (w/v) in saline-based formulations (up to 3 compounds per cassette). Blood samples (3 mice per time point) were collected at 0.0833, 0.25, 0.5, 1, 2, 4, 6, and 8 hours post-administration, and compound concentrations were measured in plasma using standard HPLC/MS analytical methods. . Tmax, Cmax, PIC, clearance, volume of distribution and oral bioavailability are estimated from plasma concentrations using non-compartmental pharmacokinetic models. Specifically, oral bioavailability (F%) is determined as F = (Dosav-AUCPo)/(DosePo-AUC)-100.

Cytotoxicity of HepG2.

Day 1. Receiving tablets C.

Cytotoxicity assays are performed in 96-well microtiter plates (Biofil, TCP 011096), the inner wells (60 inner wells surrounded by sterile water) receive 190 μl of cell count culture medium (7500 viable cells/well). One plate C containing cells is prepared for 2 compounds (8 concentrations, n=3/concentration). One background plate without cells is prepared for 6 compounds (8 concentrations, n=1/concentration)

The plates are left in place for 30 min to allow homogeneous adhesion of cells in the wells.

The plates are then incubated overnight at 37°C with 5% CO2.

Day 0: Receive Tablet A.

Stock solutions are serially diluted (semi-log dilution, 3.16x) in the appropriate 100% DMSO in a 96-well PP plate (Greiner Bio-one, 7,651,261): 79 μl of a 10,000 μ m stock solution in 100% DMSO is added to column 3, 54 μl 100% DMSO is added to columns 4-11. 25 µl of the compound stock solution is transferred from column 3 to column 4 (pipet up and down 3X, con

- 134 046887 chicks are changed after each transfer). This continues until column 10. For a positive control of doxorubicin, a stock solution at 1000 µM is used as a starting point.

Tablet A

Connection 1 10000 µM 3164.5 6 1001.4 4 316.9 1 100.2 9 31.74 10.04 3.18 100% DMSO Connection 2 10000 µM 3164.5 6 1001.4 4 316.9 1 100.2 9 31.74 10.04 3.18 100% DMSO Connection 3 10000 µM 3164.5 6 1001.4 4 316.9 1 100.2 9 31.74 10.04 3.18 100% DMSO Connection 4 10000 µM 3164.5 6 1001.4 4 316.9 1 100.2 9 31.74 10.04 3.18 100% DMSO Connection 5 10000 µM 3164.5 6 1001.4 4 316.9 1 100.2 9 31.74 10.04 3.18 100% DMSO Connection 6 10000 µM 3164.5 6 1001.4 4 316.9 1 100.2 9 31.74 10.04 3.18 100% DMSO Connection 7 10000 µM 3164.5 6 1001.4 4 316.9 1 100.2 9 31.74 10.04 3.18 100% DMSO Doxorubici n 1000 µM 316.46 100.14 31.69 10.03 3.17 1.00 0.32 100% DMSO

Receiving Tablet B.

Compounds obtained in plate A are diluted 5X in 200 µl in complete medium in a 96-well PP plate (Greiner Bio-one, 7,651,261): 200 µl of medium containing 16% Tritron X100 is added to column 2. Add 160 µl of medium to columns 3-11. 40 µl of pre-diluted compounds from plate A are transferred to columns 3-11 (pipette up and down 3X, tips changed after each transfer).

Tablet B

16% Triton X Connection 1 2000000 nM 632911.39 200288.42 63382.41 20057.72 6347.38 2008.66 635.65 20% DMSO 16% Triton X Connection 2 2000000 nM 632911.39 200288.42 63382.41 20057.72 6347.38 2008.66 635.65 20% DMSO 16% Triton X Connection 3 2000000 nM 632911.39 200288.42 63382.41 20057.72 6347.38 2008.66 635.65 20% DMSO 16% Triton X Connection 4 2000000 nM 632911.39 200288.42 63382.41 20057.72 6347.38 2008.66 635.65 20% DMSO 16% Triton X Connection 5 2000000 nM 632911.39 200288.42 63382.41 20057.72 6347.38 2008.66 635.65 20% DMSO 16% Triton X Connection 6 2000000 nM 632911.39 200288.42 63382.41 20057.72 6347.38 2008.66 635.65 20% DMSO 16% Triton X Connection 7 2000000 nM 632911.39 200288.42 63382.41 20057.72 6347.38 2008.66 635.65 20% DMSO 16% Triton X Doxorubicin 200000 nM 63291.14 20028.84 6338.24 2005.77 634.74 200.87 63.57 20% DMSO

20X solutions from plate B are diluted in plates C (with and without (background plate) cells): add 10 µl solutions of compounds from plate B compound to columns 2-11 (pipet up and down 3X, tips changed after each transfer). 2 connections are distributed into plate C three times (simplified for background plate)

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Tablet C Example

0.8% Triton X Connection 7 100000 nM 31645.57 10014.42 3169.12 1002.89 317.37 100.43 31.78 1% DMSO 0.8% Triton X Connection 7 100000 nM 31645.57 10014.42 3169.12 1002.89 317.37 100.43 31.78 1% DMSO 0.8% Triton X Connection 7 100000 nM 31645.57 10014.42 3169.12 1002.89 317.37 100.43 31.78 1% DMSO 0.8% Triton X Doxorubicin 10000 nM 3164.56 1001.44 316.91 100.29 31.74 10.04 3.18 1% DMSO 0.8% Triton X Doxorubicin 10000 nM 3164.56 1001.44 316.91 100.29 31.74 10.04 3.18 1% DMSO 0.8% Triton X Doxorubicin 10000 nM 3164.56 1001.44 316.91 100.29 31.74 10.04 3.18 1% DMSO

Plates C are incubated for 72 hours at 37°C with 5% CO ₂ .

Day 3.

µl of Alamar blue is added and incubated for 6 h at 37°C with or without CO ₂ , fluorescence is read at ex 570 nm and ex 585 nm, background corrected fluorescence measurements are converted to % scale, considering the vehicle value (1% DMSO) as 100% potency (relative measurements), or normalized using Triton X-100 values as 0% viability and vehicle values as 100% viability (normalized measurements). Relative measurements and absolute measurements are then analyzed using GraphPad Prism to obtain relative IC50 and absolute IC50 values, respectively.

The results of these activity tests are summarized in Table. 1 below.

Table 1

Number

connected ngFabl MIC50 Saso-2 Forecast F Cytotoxicity nya IC50 (µg/ml Papp A2B hep ER (%) % (patent Human We viability (nM) ) (10-6 cm/s) at 10 µm th A shi example) 5(1) < 10 < 1 22.6 0.17 100 ND 7(2) < 10 < 1 7.6 0.12 68 ND 10(3) < 10 < 1 22.6 0.22 73 ND 19(4) < 10 < 1 16 0.26 62 100 31(6) <10 <1 10.9 0.18 nd ND 32(6) < 10 < 1 10.8 0.08 42 ND 33(6) <10 <1 P,5 0.09 nd ND 39(7) <10 <1 p, z 0.17 ND ND 40(7) < 10 < 1 10.3 0.21 57 ND 41(7) <10 <1 12.2 0.09 ND ND 49(8) < 10 < 1 26 0.13 70 100 50(8) <10 <1 19.1 0.09 ND ND

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60(9) <10 < 1 3 0.07 52 ND 64(10) <10 < 1 16.3 0.21 32 ND 75(11) <10 < 1 5.9 0.13 thirty ND 80(12) <10 < 1 ND 0.25 35 ND 84(13) <10 < 1 3.29 0.40 24 ND 90(14) <10 < 1 10.7 0.03 7 ND 96(15) <10 < 1 ND 0.2 16 ND 101(16) <10 < 1 2.69 0.43 2 ND 109(17) <10 < 1 19.8 0.28 47 ND 115(18) <10 < 1 3.4 0.28 18 ND 119(19) <10 < 1 ND 0.08 ND ND 123(20) <10 < 1 2.5 0.13 26 ND 129(21) <10 < 1 ND 0.02 26 ND 134(22) <10 < 1 ND 0.43 9 ND 140(23) <10 < 1 ND 0.13 13 ND 143(24) <10 < 1 2.3 0.06 59 ND 150(25) <10 < 1 25.1 0.08 20 100 151(26) <10 < 1 21.7 0.07 39 ND 152(27) <10 < 1 2.6 0.08 38 ND 155(28) <10 < 1 ND 0.32 28 ND 159(29) <10 < 1 0.7 0.32 35 100 164(30) <10 < 1 0.1 ND 10 ND 167(31) <10 < 1 5.3 0.12 35 100 168(32) <10 < 1 19.6 0.15 51 ND 170(33) <10 < 1 22.9 0.15 98 ND

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176(36) < 10 < 1 0 0.07 44 ND 180(37) < 10 < 1 1.8 0.32 14 182(38) < 10 < 1 0.08 0.25 12 ND 183(39) < 10 < 1 ND ND 16 ND 186(40) < 10 < 1 0.4 ND 23 ND 194(41) < 10 < 1 0 ND 12 ND 198(42) < 10 < 1 3.2 0.18 54 ND 207(43) < 10 < 1 ND ND ND ND 208(44) < 10 < 1 0 0.21 ND ND 209(44) < 10 < 1 3.3 0.41 25 ND 214(45) < 10 < 1 17.9 ND 88 ND 229(46) < 10 < 1 20.7 ND 76 ND 230(46) < 10 < 1 ND ND ND ND 247(52) <10 <1 ND ND ND ND 252(53) <10 <1 ND ND ND ND 260(54) <10 <1 35 0.03 56 ND 270(55) <10 <1 5.5 0.05 ND ND 275(57) <10 <1 ND ND 51 ND 276(58) <10 <1 ND ND 43 ND 287(61) <10 <1 ND 0.34 ND ND 290(62) <10 <1 ND 0.1 ND ND

ND: not available/

Example 65/ Chlamydia trachomatis FabI inhibition assay:

FabI inhibition of Chlamydia trachomatis enzyme activity is tested by measuring the rate of NADH consumption (A absorbance at 340 nm/min) at 30°C in a 96-well plate using an automated plate reader in the presence or absence of test compounds. The assay mixture contains 100 mM MES, pH 7.0, 200 mM ammonium acetate, 4.0% glycerol, 0.02% Pluronic F-68, 25 μM crotonyl ACP, 50 μM NADH, 0.5 nM recombinant FabI protein C . trachomatis, 7.5% DMSO and test compounds at concentrations ranging from 0.17 to 100,000 nM in a final well volume of 100 µl. This dose-dependent inhibition assay is performed using 11 point serial dilution series for each test compound. IC ₅₀ values for each test compound are determined from a logistic sigmoid curve fit from the dose-dependent inhibition curves.

The results are shown in table. 2.

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table 2

Connection number (patent example) ST Fabl 1C ₅₀ (µm) 229 (46) <1 49 (8) <1 152 (27) <1 150 (25) <1 32 (6) <1 155 (28) <1 40 (7) <5 151 (26) <5 260 (54) <5 109 (17) <5 214 (45) <5 60 (9) <5 275 <5

__________________(57)_______________________________________________________________________________

The United States of America designation states: This invention was made with government support under FAIN: IDSEP160030 awarded to HHS/ASPR. The government has certain rights to an invention.

Claims (16)

1. A compound that is selected from the group of compounds represented by the general formula I where A1 represents a group selected from the groups Ac and A ₁₂ having the following structures: R ¹ - 139 046887 where the line connected to the exocyclic methylene group represents a single covalent bond formed with the nitrogen atom of formula I; A2 represents a methyl group or A1 and A2, together with the nitrogen atom to which they are bonded, form the following group A3: ^to A _z ; where the exocyclic line connected to the nitrogen atom of the bicycle represents a covalent bond between the nitrogen and the carbonyl group on the left side of formula I; and where in option (iii) Q1 represents CH2 or NH; Q2 represents CR ⁴ R ⁵ -CR ⁶ R ⁷ where the CR4R ⁵ group binds to Q1; or Q1-Q2 represents -N=CR ⁷ -CHR ⁷ - or its tautomer -NH-CR ⁷ CR ⁷ -; Q3 represents O or S; R ¹ represents a group selected from H, -NH ₂ ; R2 represents a group selected from H, -O-Ar ¹ , -O-Het ¹ , -NR ⁹ R ¹⁰ , -O-Alk ¹ where Ar ¹ represents a phenyl group which is optionally substituted by one or more groups selected from - CN, -O-C1.4-αalkyl, -O-(CH2)1.4-NR ⁹ R ¹⁰ , or where the phenyl group may bear two substituent groups on adjacent ring atoms such that these adjacent substituent groups can be bonded together to form a 5-membered heterocycle having one or two heteroatoms individually selected from N and O, where Het ¹ is an aromatic heterocycle with 5 or 6 ring atoms containing 1 or 2 heteroatoms individually selected from N, S and O, or non-aromatic partially or a fully saturated heterocycle with 6 ring atoms containing 1 heteroatom selected from N and O, wherein the Het ¹ group is optionally substituted by one or more groups individually selected from -N-alkyl, -O-C1. ₄ -αalkyl, -CN, -(CH ₂ ) ₀ . ₄ -OH; where R ⁹ is selected from H and -^^-alkyl; where R ¹⁰ is selected from H, -^l-alkyl and -C(=O)-CH ₃ ; where Alk ¹ represents an alkyl group having from 1 to 6 carbon atoms, which is linear, branched or cyclic, or a combination thereof, where the Alk ¹ group is optionally substituted by one or more groups selected from -OH, -O-alkyl; R3 represents a group selected from H, ^-PO3R3a 2, -CH2-OPO3R ^3a 2 and -CH2-OC(=O)-R ^3b ; R ^3a represents a hydrogen atom or cation suitable to form a pharmaceutically acceptable salt of either -CH2-OC(=O)-R ^3b or -CHMe-OC(=O)-R ^3b or -CMe2-OC(=O)- R ^3b , or -CH2-OC(=O)OR ^3b , or -CHMe-OC(=O)-OR ^3b , or -CMe2-OC(=O)-OR ^3b ; R ^3b represents an alkyl group having from 1 to 11 carbon atoms, which is linear, branched or cyclic, or a combination thereof, wherein the R ^3b group is optionally substituted by one or more groups independently selected from -OH and -OC ₁ . ₆ -αalkyl; R ⁴ represents a group selected from H, ^^-alkyl, CN and C ₁ . ₄ -alkylene-F; R ⁵ represents a group selected from H, ^^-alkyl, C ₁ . ₄ -alkylene-OH, C ₁ . ₄ -alkylene-OR ³ , -OH, -OPO3R ^3a 2; or R ⁴ and R ⁵ together form a cyclic group having from 4 to 6 ring members formed by methylene groups and optionally an oxygen atom; R ⁶ represents a group selected from -OH, ^^-alkyl, -OPO ₃ R ^3a 2; R ⁷ represents a group selected from H, ^^-alkyl, C ₁ . ₄ -alkylene-OH, C ₁ . ₄ -alkylene-OR ³ , ^l-alkylene^, -CN; or R ⁶ and R ⁷ together form a cyclic group having from 4 to 6 ring members formed by methylene groups and optionally an oxygen atom; where the cyclic group may optionally carry a substituent selected from -OH, -OC ₁ . ₄ -alkyl; R ⁸ represents a group selected from -O-Ar ² and -O-Het ² ; where Ar ² represents a phenyl group, which is optionally substituted by one or more groups selected from -CN, -OC ₁ . ₄ -alkyl, or where the phenyl group may bear two substituent groups on adjacent ring atoms such that these adjacent substituent groups can be bonded together to form a 5-membered heterocycle having one or two heteroatoms individually selected from N, S and O, wherein said heterocycle may optionally bear one or two substituents selected from oxo, halogen, -OC ₁ . ₄ -alkyl, ^^-alkyl and CN; and where Het ² is an aromatic heterocycle with 5 or 6 ring atoms comprising 1 or 2 heteroatoms individually selected from N, S and O, which are optionally substituted by one or more groups individually selected from -OC ₁ . ₄ -alkyl, -CN, F, ^^-alkyl, and pharmaceutically acceptable salts thereof; or - 140 046887 in option (iv) Q ¹ represents CH2 or NH; Q2 represents CR ⁴ R ⁵ -CR ⁶ R ⁷ where the CR ⁴ R ⁵ group binds to Q1; or Q1-Q2 is -N=CR ⁷ -CHR ⁷ - or its tautomer -NH-CR ⁷ =CR ⁷ -; Q3 represents O or S; R ¹ represents a group selected from H, -NH2; R2 represents a group selected from H, -O-Ar ¹ , -O-Het ¹ , -NR ⁹ R ¹⁰ , -O-Alk ¹ where Ar ¹ represents a phenyl group which is optionally substituted by one or more groups selected from - CN, -O-C1-4-alkyl, -O-(CH2)i-4-NR R' ^u . or where the phenyl group may bear two substituent groups on adjacent ring atoms such that these adjacent substituent groups can be linked together to form a 5-membered heterocycle having one or two heteroatoms individually selected from N and O, where Het ¹ represents an aromatic heterocycle with 5 or 6 ring atoms comprising 1 or 2 heteroatoms individually selected from N, S and O, or a non-aromatic partially or fully saturated heterocycle with 6 ring atoms comprising 1 heteroatom selected from N and O, where Het ¹ group is optional substituted with one or more groups individually selected from -C1-4-alkyl, -OC _1-4 -alkyl, -CN, -(CH ₂ ) _0-4 -OH; where R ⁹ is selected from H and -C1-4-alkyl; where R ¹⁰ is selected from H, -Cj-.-i-alk.ta and -C(=O)-CH3; where Alk ¹ represents an alkyl group having from 1 to 6 carbon atoms, which is linear, branched or cyclic, or a combination thereof, where the Alk ¹ group is optionally substituted by one or more groups selected from -OH, -O-C1-4- alkyl; R3 represents a group selected from H, -pO3R ^3a 2, -CH2-OPO3R ^3a 2 and -CH ₂ -OC(=O)-R ^3b ; R ^3a represents a hydrogen atom or cation suitable to form a pharmaceutically acceptable salt of either -CH2-OC(=O)-R ^3b or -CHMe-OC(=O)-R ^3b or -CMe2-OC(=O)-R ^3b or -CH ₂ -OC(=O)-OR ^3b or -CHMe-OC(=O)-OR ^3b or -CMe2-OC(=O)-OR ^3b ; R ^3b represents an alkyl group having from 1 to 11 carbon atoms, which is linear, branched or cyclic, or a combination thereof, wherein the R ^3b group is optionally substituted by one or more groups independently selected from -OH and -O-Cj-6- alkyl; R ⁴ represents a group selected from H,Cj-4-alkyl. CN and C _1-4 -alkylene-F; R ⁵ represents a group selected from H, C1-4-alkyl, C1-4-alkylene-OH, C1-4-alkylene-OR ³ , -OH, -OPO3R ^3a ₂ ; or R ⁴ and R ⁵ together form a cyclic group having from 4 to 6 ring members formed by methylene groups and optionally an oxygen atom; R ⁶ represents a group selected from H, -OH, C1-4-alkyl, -OPO3R ^3a 2; R ⁷ represents a group selected from C1-4-alkyl, C1-4-alkylene-OH, C1-4-alkylene-OR ³ , C1-4-alkylene-F, -CN; or R ⁶ and R ⁷ together form a cyclic group having from 4 to 6 ring members formed by methylene groups and optionally an oxygen atom; where the cyclic group may optionally carry a substituent selected from -OH, -OC _1-4 -alkyl; R ⁸ represents a group selected from -O-Ar ² and -O-Het ² ; where Ar ² represents a phenyl group which is optionally substituted by one or more groups selected from -CN, -OC _1-4 -alkyl, or where the phenyl group may bear two substituent groups on adjacent ring atoms such that these adjacent substituent groups may be bonded together to form a 5-membered heterocycle having one or two heteroatoms individually selected from N, S and O, wherein said heterocycle may optionally bear one or two substituents selected from oxo, halogen, -OC _1-4 -alkyl, Cj -4-alkyl and CN; and wherein Her ² is an aromatic heterocycle with 5 or 6 ring atoms comprising 1 or 2 heteroatoms individually selected from N, S and O, which are optionally substituted by one or more groups individually selected from -OC _1-4 -alkyl, -CN , F, C _1-4 -alkyl, and its pharmaceutically acceptable salts, the term alkyl refers to a saturated straight or branched hydrocarbon of 1-8 carbon atoms; a cation suitable to form a pharmaceutically acceptable salt is Na, K, Mg, Ca or the protonated form of the organic base ethanolamine, meglumine, tromethamine (2-amino-2-(hydroxymethyl)propane-1,3-diol) or deanol (2- (dimethylamino)ethanol). 2. A compound according to claim 1, wherein the term alkyl refers to a saturated straight or branched hydrocarbon of 1-6 carbon atoms. 3. The compound according to claim 1, option (iii) or (iv), where R ⁴ represents a group selected from C _1-4 -alkyl, CN and C _1-4 -alkylene-F; And R ⁵ represents a group selected from H, C _1-4 -alkyl, C _1-4 -alkylene-OH, C _1-4 -alkylene-OR ³ , -OH, -OPO3R ^3a 2; and the remaining variable groups are as defined in claim 1, option (iii) or (iv). 4. The compound according to claim 1, option (iii) or (iv), where R ⁴ represents a group selected from H, C _1-4 -alkyl, CN and C _1-4 -alkylene-F; And - 141 046887 R ⁵ represents a group selected from C _1-4 -alkyl, C _1-4 -alkylene-OH, C _1-4 -alkylene-OR ³ , -OH, -OPO, R ^ia 2; and the remaining variable groups are as defined in claim 1, option (iii) or (iv). 5. A compound according to claims 1, 2 or 3, 4, where the compound is selected from the group of compounds represented by the general formula II where R ¹ , R ² , R ³ , Q1, Q ₂ , Q ₃ have the same meanings as defined in paragraphs. 1, 2 or 3 above; and wherein the five-membered heterocycle containing _Q3 is bonded to a methyleneamide group at position 2 and a methyl group at position 3, or is bonded to a methyleneamide group at position 3 and a methyl group at position 2. 6. A compound according to claim 5, where the compound is selected from the group of compounds represented by the general formula III where R ¹ , R ² , R ³ , Q _b Q ₂ , Q3 have the same meanings as defined in paragraphs 1, 2 or 3 above. 7. A compound according to claim 6, where the compound is selected from the group of compounds represented by the general formula VI where R ¹ , R ² , R ³ , R ⁴ , R ⁵ , R ⁶ and R ⁷ have the same meanings as defined in option (iii) or (iv) according to paragraphs 1, 2 or 3 above, where in option (iii) preferably R ¹ represents H, NH ₂ ; R ² represents H; R ³ represents a group selected from H, -PO ₃ R ^3a 2, -CH2-OPO3R ^3a 2 and -CH ₂ -OC(=O)-R ^3b ; R ^3a represents a hydrogen atom or cation suitable to form a pharmaceutically acceptable salt of either -CH2-OC(=O)-R ^3b or -CHMe-OC(=O)-R ^3b or -CMe2-OC(=O)- R ^3b , or -CH ₂ -OC(=O)OR ^3b , or -CHMe-OC(=O)-OR ^3b , or -CMe2-OC(=O)-OR ^3b ; R ^3b represents an alkyl group having from 1 to 11 carbon atoms, which is linear, branched or cyclic, or a combination thereof, wherein the R ^3b group is optionally substituted by one or more groups independently selected from -OH and -O-C _1-6 -alkyl; R ⁴ represents H, C _1-4 alkyl and R ⁵ represents H, C _1-4 -alkyl, C _1-4 -alkylene-OR ³ ; R ⁶ represents C1-4-alkyl, OH, -OPO3R ^3a 2 and R ⁷ is H, C1-4-alkyl, C1-4-alkylene-OH, C1-4-alkylene-OR ³ , where R ³ is as defined above, C1-4-alkylene-H; and where in option (iv) it is preferable R ¹ represents H, NH ₂ ; R ² represents H; R ³ represents a group selected from H, -PO ₃ R ^3a 2, -CH2-OPO3R ^3a 2 and -CH ₂ -OC(=O)-R ^3b ; R ^3a represents a hydrogen atom or cation suitable to form a pharmaceutically acceptable salt of either -CH2-OC(=O)-R ^3b or -CHMe-OC(=O)-R ^3b or -CMe2-OC(=O)- R ^3b , or -CH ₂ -OC(=O)OR ^3b , or -CHMe-OC(=O)-OR ^3b , or -CMe2-OC(=O)-OR ^3b ; R ^3b represents an alkyl group having from 1 to 11 carbon atoms, which is linear, branched or cyclic, or a combination thereof, wherein the R ^3b group is optionally substituted by one or more groups independently selected from -OH and ^^^-alkyl; R ⁴ represents H, C _1-4 alkyl and R ⁵ represents H, C _1-4 -alkyl, C _1-4 -alkylene-OR ³ ; - 142 046887 R ⁶ represents H, C1-4-alkyl, OH, -OPO3R ^3a 2 and R ⁷ is C1.4-alkylene, C1-4-alkylene-OH, C1-4-alkylene-OR ³ , where R ³ is as defined above, C1-4-alkylene-F. 8. A compound according to claim 6, wherein the compound is selected from the group of compounds represented by the general formula VII where R ¹ , R ² , R ³ , R ⁶ and R ⁷ have the same meanings as defined in option (iii) or (iv) according to paragraphs 1, 2 or 3 above, where in option (iii) it is preferable R ¹ represents H, NH2; R ² represents H; R ³ represents a group selected from H, -PO ₃ R ^3a 2, -CH2-OPO3R ^3a 2 and -CH ₂ -OC(=O)-R ^3b ; R ^3a represents a hydrogen atom or cation suitable to form a pharmaceutically acceptable salt of either -CH2-OC(=O)-R ^3b or -CHMe-OC(=O)-R ^3b or -CMe2-OC(=O)- R ^3b , or -CH ₂ -OC(=O)OR ^3b , or -CHMe-OC(=O)-OR ^3b , or -CMe2-OC(=O)-OR ^3b ; R ^3b represents an alkyl group having from 1 to 11 carbon atoms, which is linear, branched or cyclic, or a combination thereof, wherein the R ^3b group is optionally substituted by one or more groups independently selected from -OH and -OC _1-6 -alkyl ; R ⁴ represents H, C _1-4 alkyl and R ⁵ represents H, C _1-4 -alkyl, C _1-4 -alkylene-OR ³ ; R ⁶ represents C1-4-alkyl, OH, -OPO3R ^3a 2 and R ⁷ is H, C1-4-alkyl, C1-4-alkylene-OH, C1-4-alkylene-OR ³ , where R ³ is as defined above, C1-4-alkylene-P; and where in option (iv) it is preferable R ¹ represents H, NH2; R ² represents H; R ³ represents a group selected from H, -PO ₃ R ^3a 2, -CH2-OPO3R ^3a 2 and -CH ₂ -OC(=O)-R ^3b ; R ^3a represents a hydrogen atom or cation suitable to form a pharmaceutically acceptable salt of either -CH2-OC(=O)-R ^3b or -CHMe-OC(=O)-R ^3b or -CMe2-OC(=O)- R ^3b , or -CH ₂ -OC(=O)OR ^3b , or -CHMe-OC(=O)-OR ^3b , or -CMe2-OC(=O)-OR ^3b ; R ^3b represents an alkyl group having from 1 to 11 carbon atoms, which is linear, branched or cyclic, or a combination thereof, where the R ^3b group is optionally substituted by one or more groups independently selected from -OH and 10-Sc-alkyl; R ⁴ represents H, C _1-4 alkyl and R ⁵ represents H, C _1-4 -alkyl, C _1-4 -alkylene-OR ³ ; R ⁶ represents H, C1-4-alkyl, OH, -OPO3R ^3a 2 and R ⁷ is C1-4-alkyl, C1-4-alkylene-OH, C1-4-alkylene-OR ³ , where R ³ is as defined above, C1-4-alkylene^. 9. A compound according to claim 5, where the compound is selected from the group of compounds represented by the general formula VIII where R ¹ , R ² , R ³ , Q1, Q ₂ , Q ₃ have the same meanings as defined in paragraphs 1, 2 or 3 above. 10. The compound according to claim 9, where the compound is selected from the group of compounds represented by the general formula XI - 143 046887 where R ¹ , R ² , R ³ , R ⁴ , R ⁵ , R ⁶ and R ⁷ have the same meanings as defined in paragraphs. 1, 2 or 3 above, where in option (iii) it is preferable R ¹ represents H; R ² represents a group selected from H, -O-Ar ¹ , -O-Het ¹ , -NH ₂ where Ar ¹ represents a phenyl group which is optionally substituted by one or more groups selected from -CN, ^^^-alkyl , or where the phenyl group may bear two substituent groups on adjacent ring atoms such that these adjacent substituent groups can be bonded together to form a 5-membered heterocycle having one or two heteroatoms individually selected from N and O, where Het ¹ represents an aromatic a heterocycle with 5 or 6 ring atoms comprising 1 or 2 heteroatoms individually selected from N, S and O, or a non-aromatic partially or fully saturated heterocycle with 6 ring atoms comprising 1 heteroatom selected from N, S and O, where Het ¹ group may optionally be substituted by one or more groups individually selected from ^^-alkyl, -O-CM-alkyl, -CN; R ³ represents a group selected from H, -PO ₃ R ^3a 2, -CH2-OPO3R ^3a 2 and -CH ₂ -OC(=O)-R ^3b ; R ^3a represents a hydrogen atom or cation suitable to form a pharmaceutically acceptable salt of either -CH2-OC(=O)-R ^3b or -CHMe-OC(=O)-R ^3b or -CMe2-OC(=O)- R ^3b , or -CH ₂ -OC(=O)OR ^3b , or -CHMe-OC(=O)-OR ^3b , or -CMe2-OC(=O)-OR ^3b ; R ^3b represents an alkyl group having from 1 to 11 carbon atoms, which is linear, branched or cyclic, or a combination thereof, wherein the R ^3b group may optionally be substituted by one or more groups independently selected from -OH and -OQ-6- alkyl; R ⁴ represents H, C _1-4 alkyl and R ⁵ represents H, C _1-4 -alkyl, C _1-4 -alkylene-OR ³ . R ⁶ represents C1-4-alkyl, OH, -OPO3R ^3a 2 and R ⁷ is H, C1-4-alkyl, C1-4-alkylene-OH, C1-4-alkylene-OR ³ , where R ³ is as defined above, C1-4-alkylene-F; where in option (iv) it is preferable R ¹ represents H; R ² represents a group selected from H, -O-Ar ¹ , -O-Het ¹ , -NH ₂ where Ar ¹ represents a phenyl group which is optionally substituted by one or more groups selected from -CN, ^^^-alkyl , or where the phenyl group may bear two substituent groups on adjacent ring atoms such that these adjacent substituent groups can be bonded together to form a 5-membered heterocycle having one or two heteroatoms individually selected from N and O, where Het ¹ represents an aromatic a heterocycle with 5 or 6 ring atoms comprising 1 or 2 heteroatoms individually selected from N, S and O, or a non-aromatic partially or fully saturated heterocycle with 6 ring atoms comprising 1 heteroatom selected from N, S and O, where Het ¹ group may optionally be substituted by one or more groups individually selected from ^^-alkyl, -O-CM-alkyl, -CN; R ³ represents a group selected from H, -PO ₃ R ^3a 2, -CH2-OPO3R ^3a 2 and -CH ₂ -OC(=O)-R ^3b ; R ^3a represents a hydrogen atom or cation suitable to form a pharmaceutically acceptable salt of either -CH2-OC(=O)-R ^3b or -CHMe-OC(=O)-R ^3b or -CMe2-OC(=O)- R ^3b , or -CH2-OC(=O)OR ^3b , or -CHMe-OC(=O)-OR ^3b , or -CMe2-OC(=O)-OR ^3b ; R ^3b represents an alkyl group having from 1 to 11 carbon atoms, which is linear, branched or cyclic, or a combination thereof, wherein the R ^3b group is optionally substituted by one or more groups independently selected from -OH and -OC _1-6 -alkyl ; R ⁴ represents H, C _1-4 alkyl and R ⁵ represents H, C _1-4 -alkyl, C _1-4 -alkylene-OR ³ . R ⁶ represents H, C1-4-alkyl, OH, -OPO3R ^3a 2 and R ⁷ is C1-4-alkyl, C1-4-alkylene-OH, C1-4-alkylene-OR ³ , where R ³ is as defined above, C1-4-alkylene-F. 11. The compound according to claim 9, where the compound is selected from the group of compounds represented by the general formula XII - 144 046887 R ^{1 K} XII where R ¹ , R ² , R ³ , R ⁶ and R ⁷ have the same meanings as defined in paragraphs 1, 2 or 3 above, where in option (iii) it is preferable R ¹ represents H; R ² represents a group selected from H, -O-Ar ¹ , -O-Het ¹ , -NH2, where Ar ¹ represents a phenyl group which is optionally substituted by one or more groups selected from -CN, -O-C1-4 -αalkyl, or where the phenyl group may bear two substituent groups on adjacent ring atoms such that these adjacent substituent groups can be linked together to form a 5-membered heterocycle having one or two heteroatoms individually selected from N and O, where Het ¹ represents an aromatic heterocycle with 5 or 6 ring atoms including 1 or 2 heteroatoms individually selected from N, S and O, or a non-aromatic partially or fully saturated heterocycle with 6 ring atoms including 1 heteroatom selected from N, S and O, wherein the Het ¹ group is optionally substituted with one or more groups individually selected from -C _1-4 -alkyl, -OC _1-4 -alkyl, -CN; R ³ represents a group selected from H, -PO ₃ R ^3a 2, -CH2-OPO3R ^3a 2 and -CH ₂ -OC(=O)-R ^3b ; R ^3a represents a hydrogen atom or cation suitable to form a pharmaceutically acceptable salt of either -CH2-OC(=O)-R ^3b or -CHMe-OC(=O)-R ^3b or -CMe2-OC(=O)- R ^3b , or -CH ₂ -OC(=O)OR ^3b , or -CHMe-OC(=O)-OR ^3b , or -CMe2-OC(=O)-OR ^3b ; R ^3b represents an alkyl group having from 1 to 11 carbon atoms, which is linear, branched or cyclic, or a combination thereof, wherein the R ^3b group is optionally substituted by one or more groups independently selected from -OH and -OC _1-6 -αalkyl ; R ⁴ represents H, C _1-4 -αalkyl and R ⁵ represents H, C _1-4 -alkyl, C _1-4 -alkylene-OR ³ ; R ⁶ represents C1-4-alkyl, OH, -OPO3R ^3a 2 and R ⁷ is H, C1-4-alkyl, C1-4-alkylene-OH, C1-4-alkylene-OR ³ , where R ³ is as defined above, C1-4-alkylene-F; where in option (iv) it is preferable R ¹ represents H; R ² represents a group selected from H, -O-Ar ¹ , -O-Het ¹ , -NH2, where Ar ¹ represents a phenyl group which may be optionally substituted by one or more groups selected from -CN, -O-C1 -4-alkyl, or where the phenyl group may bear two substituent groups on adjacent ring atoms such that these adjacent substituent groups can be bonded together to form a 5-membered heterocycle having one or two heteroatoms individually selected from N and O, where Het ¹ is an aromatic heterocycle with 5 or 6 ring atoms comprising 1 or 2 heteroatoms individually selected from N, S and O, or a non-aromatic partially or fully saturated heterocycle with 6 ring atoms containing 1 heteroatom selected from N, S and O, wherein the Het ¹ group is optionally substituted with one or more groups individually selected from -C _1-4 -alkyl, -OC _1-4 -alkyl, -CN; R ³ represents a group selected from H, -PO3R ^3a 2, -CH2-OPO3R ^3a 2 and -CH2-OC(=O)-R ^3b ; R ^3a represents a hydrogen atom or cation suitable to form a pharmaceutically acceptable salt of either -CH2-OC(=O)-R ^3b or -CHMe-OC(=O)-R ^3b or -CMe2-OC(=O)- R ^3b , or -CH2-OC(=O)OR ^3b , or -CHMe-OC(=O)-OR ^3b , or -CMe2-OC(=O)-OR ^3b ; R ^3b represents an alkyl group having from 1 to 11 carbon atoms, which is linear, branched or cyclic, or a combination thereof, wherein the R ^3b group is optionally substituted by one or more groups independently selected from -OH and -OC _1-6 -alkyl ; R ⁴ represents H, C _1-4 -alkyl and R ⁵ represents H, C _1-4 -alkyl, C _1-4 -alkylene-OR ³ . R ⁶ represents H, C1-4-alkyl, OH, -OPO3R ^3a 2 and R ⁷ is C1-4-alkyl, C1-4-alkylene-OH, C1-4-alkylene-OR ³ , where R ³ is as defined above, C1-4-alkylene-F. 12. A compound according to claim 1, where the compound is selected from the group of compounds represented by the general formula XIII - 145 046887 where R ³ , R ⁸ , Q1, Q2 have the same meanings as defined in paragraphs. 1, 2 or 3 above. 13. The compound according to claim 12, where the compound is selected from the group of compounds represented by the general formula XV where R ³ , R ⁴ , R ⁵ , R ⁶ , R ⁷ , R ⁸ have the same meanings as defined in paragraphs. 1, 2 or 3 above, where in option (iii) it is preferable R ³ represents a group selected from H, -PO ₃ R ^3a 2, -CH2-OPO3R ^3a 2 and -CH ₂ -OC(=O)-R ^3b ; R ^3a represents a hydrogen atom or cation suitable to form a pharmaceutically acceptable salt of either -CH2-OC(=O)-R ^3b or -CHMe-OC(=O)-R ^3b or -CMe2-OC(=O)- R ^3b , or -CH ₂ -OC(=O)OR ^3b , or -CHMe-OC(=O)-OR ^3b , or -CMe2-OC(=O)-OR ^3b ; R ^3b represents an alkyl group having from 1 to 11 carbon atoms, which is linear, branched or cyclic, or a combination thereof, wherein the R ^3b group is optionally substituted by one or more groups independently selected from -OH and -OC _1-6 -alkyl ; R ⁴ represents H, C _1-4 alkyl and R ⁵ represents H, C _1-4 -alkyl, C _1-4 -alkylene-OR ³ ; R ⁶ represents C1-4-alkyl, OH, -OPO3R ^3a 2 and R ⁷ is H, C1-4-alkyl, C1-4-alkylene-OH, C1-4-alkylene- ^OR3 , where ^R3 is as defined above, C1-4-alkylene^; And R ⁸ represents a group selected from -O-Ar ² and -O-Het ² , where Ar ² represents a phenyl group that is optionally substituted by one or more groups selected from -CN, ^-Cy^alkyl, or where the phenyl group may bear two substituent groups on adjacent ring atoms such that these adjacent substituent groups can be linked together to form a 5-membered heterocycle having one or two heteroatoms individually selected from N, S and O, wherein said heterocycle may optionally bear one or two substituents selected from oxo, F, -OC _1-4 -alkyl, C _1-4 -alkyl, CN; and where Het ² represents an aromatic heterocycle with 5 or 6 ring atoms comprising 1 or 2 heteroatoms individually selected from N, S and O, which are optionally substituted by one or more groups individually selected from -O-Ci-4-alkyl, - CN, F, C1 _-4 -alkyl; where in option (iv) it is preferable R ³ represents a group selected from H, -PO ₃ R ^3a 2, -CH2-OPO3R ^3a 2 and -CH ₂ -OC(=O)-R ^3b ; R ^3a represents a hydrogen atom or cation suitable to form a pharmaceutically acceptable salt of either -CH2-OC(=O)-R ^3b or -CHMe-OC(=O)-R ^3b or -CMe2-OC(=O)- R ^3b , or -CH2-OC(=O)OR ^3b , or -CHMe-OC(=O)-OR ^3b , or -CMe2-OC(=O)-OR ^3b ; R ^3b represents an alkyl group having from 1 to 11 carbon atoms, which is linear, branched or cyclic, or a combination thereof, wherein the R ^3b group is optionally substituted by one or more groups independently selected from -OH and -OC _1-6 -alkyl ; R ⁴ represents H, C _1-4 alkyl and R ⁵ represents H, C _1-4 -alkyl, C _1-4 -alkylene-OR ³ ; R ⁶ represents H, C1-4-alkyl, OH, -OPO3R ^3a 2 and R ⁷ is C1-4-alkyl, C1-4-alkylene-OH, C1-4-alkylene-OR ³ , where R ³ is as defined above, C1-4-alkylene^; And R ⁸ represents a group selected from -O-Ar ² and -O-Het ² , where Ar ² represents a phenyl group that is optionally substituted by one or more groups selected from -CN, ^-Cl^alkyl, or where the phenyl group may bear two substituent groups on adjacent ring atoms such that these adjacent substituent groups can be bonded together to form a 5-membered heterocycle having one or two heteroatoms individually selected from N, S and O, wherein said heterocycle may optionally bear one or two substituents selected from oxo, F, -OC _1-4 -alkyl, C _1-4 -alkyl, CN; and where Het ² represents an aromatic heterocycle with 5 or 6 ring atoms, comprising 1 or 2 heteroatoms individually selected from N, S and O, which may optionally be substituted by one or more groups individually selected from -O-C _1-4 - alkyl, -CN, F, C _1-4 -alkyl. 14. The compound according to claim 12, where the compound is selected from the group of compounds represented by the general formula XVI where R ³ , R ⁴ , R ⁵ , R ⁶ , R ⁷ , R ⁸ have the same meanings as defined in paragraphs 1, 2 or 3 above, where in option (iii) it is preferable R ³ represents a group selected from H, -PO ₃ R ^3a 2, -CH2-OPO3R ^3a 2 and -CH ₂ -OC(=O)-R ^3b ; R ^3a represents a hydrogen atom or cation suitable to form a pharmaceutically acceptable salt of either -CH2-OC(=O)-R ^3b or -CHMe-OC(=O)-R ^3b or -CMe2-OC(=O)- R ^3b , or -CH2-OC(=O)OR ^3b , or -CHMe-OC(=O)-OR ^3b , or -CMe2-OC(=O)-OR ^3b ; R ^3b represents an alkyl group having from 1 to 11 carbon atoms, which is linear, branched or cyclic, or a combination thereof, wherein the R ^3b group is optionally substituted by one or more groups independently selected from -OH and -OC _1-6 -alkyl ; R ⁴ represents H, C _1-4 alkyl and R ⁵ represents H, ^ alkyl, C _1-4 -alkylene-OR ³ ; R ⁶ represents C1-4-alkyl, OH, -OPO3R ^3a 2 and R ⁷ represents H, Ci-4-alkyl, ^n-alkylene1^ C1-4-alkylene-OR ³ , where R ³ is as defined above, C1-4-alkylene-F; R ⁸ represents a group selected from -O-Ar ² and -O-Het ² , where Ar ² represents a phenyl group which may be optionally substituted by one or more groups selected from -CN, -OC _1-4 -alkyl, or wherein the phenyl group may bear two substituent groups on adjacent ring atoms such that these adjacent substituent groups may be bonded together to form a 5-membered heterocycle having one or two heteroatoms selected from N, S and O, wherein said heterocycle may optionally bear one or two substituents individually selected from oxo, F,-O-CM-alkyl, ^-alkyl, CN; where Het ² represents an aromatic heterocycle with 5 or 6 ring atoms, including 1 or 2 heteroatoms individually selected from N, S and O, which are optionally substituted by one or more groups individually selected from -OC _1-4 -alkyl, -CN, F, ^-alkyl; where in (iv) it is preferable R ³ represents a group selected from H, -PO ₃ R ^3a 2, -CH2-OPO3R ^3a 2 and -CH ₂ -OC(=O)-R ^3b ; R ^3a represents a hydrogen atom or cation suitable to form a pharmaceutically acceptable salt of either -CH2-OC(=O)-R ^3b or -CHMe-OC(=O)-R ^3b or -CMe2-OC(=O)- R ^3b , or -CH2-OC(=O)OR ^3b , or -CHMe-OC(=O)-OR ^3b , or -CMe ₂ -OC(=O)-OR ^3b ; R ^3b represents an alkyl group having from 1 to 11 carbon atoms, which is linear, branched or cyclic, or a combination thereof, wherein the R ^3b group is optionally substituted by one or more groups independently selected from -OH and 10-alkyl; R ⁴ represents H, C _1-4 alkyl and R ⁵ represents H, C _1-4 -alkyl, C _1-4 -alkylene-OR ³ ; R ⁶ represents H, C1-4-alkyl, OH, -OPO3R ^3a 2 and R ⁷ is C1-4-alkyl, C1-4-alkylene-OR ³ , where R ³ is as defined above, C1-4-alkylene-F; R ⁸ represents a group selected from -O-Ar ² and -O-Het ² , where Ar ² represents a phenyl group that is optionally substituted by one or more groups selected from -CN, -OC _1-4 -alkyl, or where phenyl group may bear two substituent groups on adjacent ring atoms such that these adjacent substituent groups can be linked together to form a 5-membered heterocycle having one or two heteroatoms selected from N, S and O, wherein said heterocycle may optionally bear one or two substituents individually selected from oxo, F, 10^-alkyl, C _1-4 -alkyl, CN; and where Het ² represents an aromatic heterocycle with 5 or 6 ring atoms, including 1 or 2 heteroatoms individually selected from N, S and O, which may optionally be substituted by one or more groups individually selected from -OC _1-4 -alkyl, -CN, F, C _1-4 -alkyl. - 147 046887 15. A pharmaceutical composition containing a compound according to any one of claims 1 to 14 and a pharmaceutically acceptable carrier. 16. Use of a compound according to any one of claims 1 to 14 or a pharmaceutical composition according to claim 15 for the treatment of a bacterial infection selected from infections caused by N. gonorrhoeae, Chlamydia trachomatis or a combination thereof.