OA18890A

OA18890A - New aminoacid derivatives, a process for their preparation and pharmaceutical compositions containing them

Info

Publication number: OA18890A
Application number: OA1201700512
Authority: OA
Inventors: Olivier Geneste; James Edward Paul Davidson; James Brooke MURRAY; Alain Bruno; l-Jen CHEN; Françoise PERRON-SIERRA; Andras Kotschy; Szabolcs SIPOS; Attila Paczal; Balazs Balint; Zoltan Szabo; Marton Csékei; Levente Ondi; Gabor Radics; Agnes Proszenyak; Zoltan Szlavik
Original assignee: Les Laboratoires Servier; Vernalis (R&D) Limited
Priority date: 2015-06-23
Filing date: 2016-06-22
Publication date: 2019-09-30

Abstract

Compounds of formula (I):

Description

The present invention relates to new aininoacid dérivatives, to a process for their préparation and to pharmaceutical compositions containing them.

The compounds of the present invention are new and hâve very valuable pharmacological characteristics in the field of apoptosis and cancerology.

Apoptosis, or programmed cell death. is a physiological process that is crucial for embryonic development and maintenance of tissue homeostasis.

Apoptotic-type cell death involves morphological changes such as condensation of the nucléus, DNA fragmentation and also biochemical phenomena such as the activation of caspases which cause damage to key structural components of the cell, so inducing its disassembly and death. Régulation of the process of apoptosis is complex and involves the activation or repression of several intracellular signalling pathways (Cory S. et al., Nature Review Cancer 2002, 2, 647-656).

Deregulation of apoptosis is involved in certain pathologies. Increased apoptosis is associated with neurodegenerative diseases such as Parkinson s disease, Alzheimer s disease and ischaemia. Conversely, déficits in the implémentation of apoptosis play a significant rôle in the development of cancers and their chemoresistance, in auto-immune diseases, inflammatory diseases and viral infections. Accordingly, absence of apoptosis is one of the phenotypic signatures of cancer (Hanahan D. et al., Cell 2000, 100, 57-70).

The anti-apoptotic proteins of the Bcl-2 family are associated with numerous pathologies. The involvement of proteins of the Bcl-2 family is described in numerous types of cancer, such as colon cancer, breast cancer, small-cell lung cancer, non-small-cell lung cancer, bladder cancer, ovarian cancer, prostate cancer, chronic lymphoid leukaemia, lymphoma, myeloma, acute myeloid leukemia, pancreatic cancer, etc. Overexpression of the antiapoptotic proteins of the Bcl-2 family is involved in tumorigenesis, in résistance to chemotherapy and in the clinical prognosis of patients affected by cancer. Notably, Mcl-l, an anti-apoptotic Bcl-2 family member, is overexpressed in varions types of cancer (Beroukhim R. et al.. Nature 2010, 899-905). There is, therefore, a therapeutic need for compounds that inhibit the anti-apoptotic activity of the proteins of the Bcl-2 family.

ORIGINAL

-2ln addition to being new, the compounds of the present invention hâve pro-apoptotic properties making it possible to use them in pathologies involving a detect in apoptosis, such as, for example, in the treatment of cancer and of immune and auto-immune diseases.

The present invention relates more especially to compounds of formula (I):

(D wherein:

♦ A represents the group

in which 1 is linked to the -NH- group and 2 is linked to the aromatic ring,

E represents a cycloalkyl group. a heterocycloalkyl group, an aryl group or a heteroaryl group,

X represents a nitrogen atom or a C-IU group.

Y represents a nitrogen atom or a C-R3 group.

Ri represents a halogen atom, a linear or branched (C|-C(,)alkyl group, a linear or branched (C2-C₆)alkenyl group, a linear or branched (C2-C₆)alkynyl group, a linear or branched (Ci-C₆)polyhaloalkyl group, a hydroxy group, a hydroxy(C|-C₆)alkyl group, a linear or branched (Cj-C₆)alkoxy group, -S-(C i-Côfalkyl, a cyano group, a nitro group, -alkyl(Co-C6)-NR₉R₉\ -O-alkyl(C|-C(,)-NR₉R₉’, -O-alkyl(Ci-C6)-R)0, -C(O)-OR₉, -O-C(O)-R₉, -C(O)-NR₉R₉', -NR₉-C(O)-R₉’, -NR₉-C(O)-OR₉’, -alkyl(C[-C6)-NR₉-C(O)-R₉’, -SO₂-NR₉R₉’, -SO₂-alkyl(C|-C₆).

- R₂. R3, Ki and R₅ îndependently of one another represent a hydrogen atom, a

ORIGINAL

- 3 halogen atom. a linear or branched (Ci-Côjalkyl group, a linear or branched (C₂-Cô)alkenyl group, a linear or branched (C₂-C6)alkynyl group, a linear or branched (C_t-C₆)polyhaloalkyl group, a hydroxy group, a hydroxy(Ci-C₆)alkyl group, a linear or branched (Ci-Côjalkoxy group, -S-(Ci-C6)alkyl, a cyano group, a nitro group, -alkyl(Co-C6)-NR₉R₉’, -O-alkyl(C|-C6)-NR₉R₉'’, -0-alkyl(C|-C6)-Rio, -C(O)-OR₉, -O-C(O)-R₉, -C(O)-NR₉R₉\ -NR₉-C(O)-R₉’, -NR₉-C(O)-OR₉\ -a!kyl(C|-C₆)-NR₉-C(O)-R₉’, -SO₂-NR₉R₉ ^?, or -SO₂-alkyl(Cj-C₆), or the substituents ofthe pair (R_l? R₂) form together with the carbon atoms carrying them an aromatic or non-aromatic ring composed of from 5 to 7 ring members, which may contain from l to 3 heteroatoms selected from oxygen, sulphur and nitrogen. it being understood that resulting ring may be substituted by from l to 2 groups selected from halogen, linear or branched (CrCôjalkyl, -aikyl(Co-C6)-NR₉R₉\ -NRj ₍Ri Γ, -alkyl(C_Q-C₆)-Cyi, or oxo,

Rb represents a hydrogen atom, a halogen atom, a linear or branched (C|-Cf,)alkyl group, a linear or branched (C₂-C6)alkenyl group, a linear or branched (C₂-C₆)alkynyl group, a linear or branched (Ci-C₆)polyhaloalkyl group, a hydroxy group, a linear or branched (Ci-Cô)alkoxy group, -S-(Ci-Côlalkyl, a cyano group, a nitro group, -alkyl(Co-C6)-NR₉R₉\ -O-Cyi, -alkyl(Co-C₆)-Cyi, -alkenyl(C₂-C6)-Cyi, -alkynyl(C₂-C₆)-Cyi, -0-alkyl(C|-C6)-Rio, -C(O)-OR₉, -O-C(O)-R₉.

-C(O)-NR₉R₉', -NR₉-C(O)-R₉’, -NR₉-C(O)-OR₉’, -alkyl(Ci-C₆)-NR₉-C(O)-R₉’. -SO₂-NR₉R₉’, or -SO₂-alkyl(Ci-C₆),

R7 represents a hydrogen atom, a linear or branched (Ci-Cs)alkyl group, a -CHR_nRb group, an aryl group, a heteroaryl group, an arylalkyftCi-Cô) group, or a heteroarylalkyl(Ci-Cf,) group,

Rs represents a linear or branched (Cj-C^alkyl group. a linear or branched (C₂-C6)alkenyl group, a linear or branched (C₂-C6)alkynyl group, -Cy₂, a halogen atom, a cyano group, -C(O)-Rn, or -C(O)-NRi|Rn’,

R₉ and R₉’ independently of one another represent a hydrogen atom, a linear or branched (Ci-Cùjalkyl group, or the substituents of the pair (R₉, R₉’) form together with the nitrogen atom carrying them an aromatic or non-aromatic ring composed of from 5 to 7 ring members, which may contain in addition to the nitrogen atom from l to 3

ORIGINAL

-4heteroatoms selected from oxygen, sulphur and nitrogen, it being understood that the nitrogen in question may be substituted by a group representing a hydrogen atom, or a linear or branched (Ci-C₆)alkyI group,

Rio represents -Cy₃, -Cy₃-alkyl(Co-C6)-Cy4, -C(O)-NR₉R.9’, -NR9R9’, -OR9, -NRç-C(O)-R₉’, -O-alkyl(C_l-C₆)-OR₉, -SO2-R9, -C(O)-OR₉.or -NH-C(O)-NH-R₉,

- R_M and Ru’ independently of one another represent a hydrogen atom or an optionally substituted linear or branched (C ]-Cô)alkyl group,

Ru represents a hydrogen atom, a hydroxy group, or a hydroxy(C|-Cé)alkyl group, R_a represents a hydrogen atom or a linear or branched (Ci-Cô)alkyl group, Rb represents a -O-C(O)-O-R_t group, a -O-C(O)-NR_cR_c’ group, or a -O-P(O)(OR_C)₂group,

R_c and R_e’ independently of one another represent a hydrogen atom, a linear or branched (C_rC₈)alkyl group, a cycloalkyl group, a (Ci-C₆)alkoxy(C]-C6)alkyl group, a (Ci-C6)alkoxycarbonyl(Ci-C6)alkyl group, or the substituents of the pair (R_€, R_c’) form together with the nitrogen atom carrying them a non-aromatic ring composed of from 5 to 7 ring members, which may contain in addition to the nitrogen atom from l to 3 heteroatoms selected from oxygen and nitrogen, it being understood that the nitrogen in question may be substituted by a group representing a linear or branched (Ci-C₆)alkyl group.

- Cyi, Cy₂, Cy₃ and Cy₄ independently of one another, represent a cycloalkyl group, a heterocycloalkyl group, an aryl group or a heteroaryl group, n is an integer equal to 0, l or 2, it being understood that:

- “aryl” means a phenyl, naphlhyl, biphenyl group, “heteroaryl” means any mono- or bi-cyclic group composed of from 5 to I0 ring members, having at least one aromatic moiety and containing from l to 3 heteroatoms selected from oxygen, sulphur and nitrogen, “cycloalkyl” means any mono- or bi-cyclic non-aromatic carbocyclic group containing from 3 to 10 ring members, “heterocycloalkyl” means any mono- or bi-cyclic non-aromatic carbocyclic group containing from 3 to 10 ring members, and containing from l to 3 heteroatoms

ORIGINAL

-5selected from oxygen, sulphur and nitrogen, which may include fused, bridged or spiro ring Systems, io it being possible for the aryl, heteroaryl, cycloalkyl and heterocycloalkyl groups so defined and the alkyl, alkenyl, alkynyl, alkoxy groups, to be substituted by from l to 4 groups selected from optionally substituted linear or branched (Ci-C&)alkyl, optionally substituted linear or branched (C₂-C₆)alkenyl, optionally substituted linear or branched (Ci-Côjalkynyl, optionally substituted linear or branched (C|-Cô)alkoxy, optionally substituted (C,-C₆)alkyl-S-, hydroxy, hydroxy(Ci-C₆)alkyl, oxo (or A-oxide where appropriate), nitro, cyano, -C(O)-OR’, -O-C(O)-R’, -C(O)-NR R”, -O-C(O)-NR R”, -NR’R’L -(C=NR’)-0R”, -O-P(O)(OR’)₂, -0-P(0)(0’M⁺)₂, linear or branched (C|-C₆)polyhaloalkyl, trifluoromelhoxy, halogen, or an aldohexose of formula:

in which each R’ is indépendant;

it being understood that R’ and R” independently of one another represent a hydrogen atom or an optionally substituted linear or branched (Cj-C₆)alkyl group and M^r represents a pharmaceutically acceptable monovalent cation, their enantiomers, diastereoisomers and atropisomers, and addition salts thereof with a pharmaceutically acceptable acid or base.

Advantageously, the présent invention relates to compounds of formula (I) wherein:

- Ri and R₂ independently of one another represent a halogen atom, a linear or branched (C|-C₆)alkyl group, a hydroxy group, a hydroxy(C|-C₆)alkyl group, a linear or branched (C|-C6)alkoxy group, or the substituents of the pair (R_b R₂) form together with the carbon atoms carrying them an aromatic ring composed of from 5 to 7 ring members, which may contain

ORIGINAL

- 6 from l to 3 nitrogen atoms, it being understood that resuking ring may be substituted by from l to 2 groups selected from halogen, linear or branched (C|-C₆)alkyl, or -alkyl(Co-C₆)-NR₉R9’,

- R₃ represents a hydrogen atom, a halogen atom, a linear or branched (Ci-C₆)alkyl group, a hydroxy group, a linear or branched (Ci-C₆)alkoxy group, or -O-alkyl(C_rC₆)-NR₉R₉’,

- R4 and R₅ independently of one another represent a hydrogen atom, a halogen atom, a linear or branched (C|-C₆)alkyl group, a hydroxy group, a linear or branched (C_rC6)alkoxy group,

R_f, represents a hydrogen atom, a halogen atom, a linear or branched (C|-C₆)alkyl group, a linear or branched (Cj-Cojpolyhaloalkyl group, a hydroxy group, a linear or branched (Ci-C₆)alkoxy group, a cyano group, a nitro group, -alkyl(C₀-C6)-NR₉R9’, -alkyl(C₀-C₆)-Cy_b -O-alkyl(C_rC₆)-Ri₀ or -C(O)-NR₉R₉’, R7 represents a hydrogen atom, a linear or branched (Ci-C_g)alkyl group, a -CHR_aRb group, or a heteroarylalkyl(Ci-Cô) group,

Rs represents a linear or branched (Ci-Cô)alkyl group, a linear or branched (C₂-C6)alkenyl group, a linear or branched (C₂-C6)alkynyl group, -Cy₂, a halogen atom, or -C(O)-Rh,

R₉ and R₉’ independently of one another represent a hydrogen atom. a linear or branched (C]-C₆)alkyl group, or the substituents of the pair (R₉, R₉ ^?) form together with the nitrogen atom carrying them a non-aromatic ring composed of from 5 to 7 ring members, which may contain in addition to the nitrogen atom from l to 3 heteroatoms selected trom oxygen and nitrogen, it being understood that the nitrogen in question may be substituted by a group representing a linear or branched (Ci-Cb)alkyl group,

Rio represents -Cy₃ or -Cy₃-alkyl(Co-C_f,)-Cy₄,

Rii represents a linear or branched (Ci-Cojalkyl group, it being possible for the aryl, heteroaryl. cycloalkyl and heterocycloalkyl groups so deiïned and the alkyl, alkenyl, alkynyl, alkoxy groups, to be substituted by from l to 4 groups selected from optionally substituted linear or branched (Cj-Cojalkyl, optionally substituted linear or branched (Cj-C₆)alkoxy, hydroxy, hydroxy(C_rC₆)alkyl, oxo (or N-oxide where

ORIGINAL

-7appropriate). -C(O)-OR', -C(0)-NR’R -0-C(0)-NR'R”. -NR’R, -O-P(O)(OR’)₂, -0-P(0)(0‘M⁺)₂, linear or branched (C]-C₆)polyhaloalkyl, halogen, or an aldohexose of formula:

in which each R’ is independent;

it being understood that R’ and R” independently of one another represent a hydrogen atom or an optionally substituted linear or branched (C]-C(>)alkyl group and M represents a pharmaceutically acceptable monovalent cation.

Among the pharmaceutically acceptable acids there may be mentioned, without implying 10 any limitation, hydrochloric acid, hydrobromic acid, sulphuric acid, phosphonic acid, acetic acid, trifluoroacetic acid, lactic acid, pyruvic acid, malonic acid, succinic acid, glutaric acid, fumaric acid, tartane acid, maleic acid, citric acid, ascorbic acid, oxalic acid, methanesulphonic acid, camphoric acid etc.

Among the pharmaceutically acceptable bases there may be mentioned, without implying 15 any limitation, sodium hydroxide, potassium hydroxide, trîethylamine, re/7-butylamine etc.

More especially, compounds of formula (I) to which préférence is given are compounds wherein n is an integer equal to l.

In another embodiment of the invention, an advantageous possibility consists of compounds of formula (I-a):

ORIGINAL

(I-a) wherein A, E, R_h R₂, R₅, R₆, R?, R12, X. Y and n are as defined for formula (I).

Atropisomers are stereoisomers arising because ot hindered rotation about a single bond, where energy différences due to steric strain or other contributors create a barrier to rotation that is high enough to allow for isolation of individual conformers. For compounds according to the invention, atropisomers are as follows:

Preferred atropisomer is (55_a) when X represents a C-R4 group and Y represents a C-R₃ group.

I0 Advantageously, at least one of the groups selected from R₂, R₃, R4 and R5 does not represent a hydrogen atom.

Preferably, R|₂ represents a hydrogen atom, a hydroxymethyl group or a hydroxyethyl group. More preferably, R_l2 represents a hydrogen atom.

In the preferred compounds of the invention. R| represents a linear or branched

I5 (C]-C6)alkyl group or a halogen atom. More preferably. Ri represents a methyl group, an

ORIGINAL

-9ethyl group, a bromine atom or a chlorine atom. Even more preferably, R, represents a methyl group.

Advantageously, R? represents a halogen atom, a hydroxy group, a linear or branched (C|-C₆)alkoxy group. More preferably, R₂ represents a methoxy group, a hydroxy group, a 5 fluorine atom, a bromine atom or a chlorine atom. Even more preferably, R₂ represents a chlorine atom.

In some preferred embodiment of the invention, when the substituents of the pair (Ri, R₂) form together with the carbon atoms carrying them an aromatic ring,

wherein Ru represents a hydrogen atom, a linear or branched (Cj-CôJalkyl group or -alkyl(Co-C₆)-NR₉R₉’ in which R₉ and R₉ ^? are as defined for formula (l), and R_!4represents a hydrogen atom, a halogen atom or a linear or branched (Ci-Cb)alkyl group.

I5 R|3 represents preferably a hydrogen atom, a methyl group or -(CH₂)_m-NR₉R₉’ in which m is an integer equal to 2 or 3 and, R₉ and R₉ ^? represent a methyl group or the substituents of the pair (R₉, R₉’) form together with the nitrogen atom carrying them a pyrrolidinyl, a piperidinyl, a morpholinyï or a 4-methyl-piperazin-l -yl group.

ORIGINAL

- ΙΟR_l4 represents advantageously a hydrogen atom, a bromine atom. an iodine atom, a chlorine atom or a methyl group. R|.| is preferably substituted at β position from the nitrogen atom.

Preferably, X represents a C-R4 group. In a preferred embodiment of the invention, Y represents a C-R₃ group. R₃ advantageously represents a hydrogen atom, a linear or branched (C_rC₆)alkoxy group or -O-alkyKCi-Côj-NRçRy'. R4 preferably represents a hydrogen atom.

In some preferred embodiment of the invention,

wherein R_ls R?, R9 and R</ are as defined for formula (1).

In the preferred compounds ofthe invention.

wherein R₉ and R₉’ are as defined for formula (I).

Rs preferably represents a hydrogen atom.

ORIGINAL

- Il In an advantageous embodiment, the substituents of the pair (R.[, R,) are identical and the substituents of the pair (R?, R-ι) are identical. In the preferred compounds of the invention, the substituents of the pair (Ri, R₅) are identical and represent a (C|-C₆)alkyl group, preferably a methyl group, whereas the substituents of the pair (R₂, Ri) are identical and represent a halogen atom, preferably a chlorine atom, or a hydrogen atom.

In another embodiment of the invention, E represents a phenyl group. a pyridin-2-yl, a cyclohexyl group, a pyrazol-l-yl group, a cyclopentyl group, an indol-4-yl group, a cyclopropyl group, a pyridin-3-yl group, an indol-3-yl group, a naphth-l-yl group, an imidazol-4-yl group or a pyridin-4-yl group. Advantageously, E represents a phenyl group.

In the preferred compounds of the invention, R₍, represents a hydrogen atom; a fluorine atom; a chlorine atom; a bromine atom; a methyl group; a trifluoromethyl group; a hydroxy group; a methoxy group; a linear (CrC&)alkoxy group substituted by halogen atoms, _a -C(O)-NR’R” group or a -NR’R” group; a cyano; a nitro group; an aminomethyl group; a benzyl group; -O-alkyRCrCôj-Rio; -C(O)-NR₉R<. Preferably, Ri, represents a methoxy group, a 2,2,2-trifluoroethoxy group or -0-alkyl(C_rC6)'Rio.

In another embodiment of the invention, an advantageous possibility consists of compounds of formula (l-b):

wherein Ri, R₂, R5, R₍₎, R7, R12, X Y, A and n are as defined for formula (I).

In another embodiment of the invention, an advantageous possibility consists of

ORIGINAL

- 12compounds of formula (I-c):

wherein R₆, R₇, R₉. RJ, R12 and A are as defined for formula (J).

Preferably. R7 represents a hydrogen atom, a -CHR_aRb group, an optionally substituted linear or branched (Ci-Cs)alkyl group, or a heteroarylalkyl(Ci-Cb) group. Preterably, R₇ represents a -CHR_:iR_h group in which R_a represents a hydrogen atom or a methyl group and R_b represents a -O-C(O)-O-(Ci-C_g)alkyl group; a -O-C(O)-O-cycloalkyl group; a -O-C(O)-NR_cRJ group, in which R_c and RJ independently of one another represent a hydrogen atom, a linear or branched (Ci-Cs)alkyl group, a (C|-C6)alkoxy(Ci~C6)alkyl group, a (Ci-C₆)alkoxycarbonyl(Ci-C6)alkyl group, or the substituents of the pair (R_c, R_c ) form together with the nitrogen atom carrying them a non-aromatic ring composed of from 5 to 7 ring members, which may contain in addition to the nitrogen atom from l to 3 heteroatoms selected from oxygen and nitrogen; or a -O-P(O)(OH)2 group. Preferred R₇groups are as follows: hydrogen; methyl; ethyl; (5-methyl-2-oxo-l,3-dioxol-4-yl)methyl; a -CHR_aR_b group in which R_a represents a methyl group and R_b represents _a -O-C(O)-O-CH₂CH₃ group or a -O-C(O)-N(CH₃)₂ group. Even more preferably, R₇represents hydrogen.

In the preferred compounds of the invention, Rs represents a linear or branched (CT-Célalkynyl group, an aryl group or a heteroaryl group, More preferably, Rs represents a prop-l-yn-l-yl group, a phenyl group or a furan-2-yl group. In a more preferred embodiment, Rg represents a prop-l-yn-l-yl group, a 4-fluorophenyl group 01 a

5-fluorofuran-2-yl group. Even more preferentially, R₈ represents a 4-fluorophenyl group.

ORIGINAL

- 13 In the preferred compounds of the invention, Rç and R<f independently of one another represent a linear or branched (Ci-Cô)alkyl group, or the substituents of the pair (R₉, R₉’) form together with the nitrogen atom carrying them a non-aromatic ring composed of from 5 to 7 ring members, which may contain in addition to the nitrogen atom from l to 3 heteroatoms selected from oxygen and nitrogen, it being understood that the nitrogen in question may be substituted by a linear or branched (C|-C6)alkyl group. More preferably, R₉ and R₉’ represent a methyl group, or the substituents of the pair (R9, R9’) form together a 4-methyl-piperazinyl group.

Advantageously, Rio represents -Cya or -Cya-alkyl(Co-C6)-Cy₄. Preferably, Rio represents -Cy₃ or -Cy₃-Cy₄.

Cya preferably represents a cycloalkyl group, particularly, a cyclopentyl group. In a preferred embodiment, Cy3 represents an aryl group, particularly, a phenyl group. Advantageously, Cy3 represents a heteroaryl group, particularly, a pyrimidinyl group, a pyrazolyl group or a pyridinyl group. More preferably, Cya represents a pyrimidin-4-yi group, a pyrazol-5-yl group or a pyridin-2-yl group. In the preferred compounds of the invention, Cya represents a pyrimidin-4-yl group. In another embodiment of the invention, Cy3 represents a heteroaryl group which is substituted by an optionally substituted linear or branched (C|-C₆)alkyl group, an optionally substituted linear or branched (C|-C₆)alkoxy group or a linear or branched (Ci-C6)polyhaloalkyl group. Preferably, Cya represents a heteroaryl group which is substituted by a 2,2,2-trifluoroethoxy group, a 2-methoxyethyl group, an ethoxy group; a Ze/7-butyl group, an ethyl group, a zz-butyl group, a 2,2,2-trifluoroethyl group or a methyl group.

Cy₄ preferably represents a phenyl group, a pyridinyl group, a pyridazinyl group, a pyrazinyl group, a pyrimidinyl group or a morpholinyl group. More preferably, Cy₄represents a phenyl group.

Other compounds of the invention to which preference is given are those wherein,

ORIGINAL

Rio represents

in which p is an integer equal to 0 or l and R|s represents a hydrogen atom, a hydroxy group, an optionally substituted linear or branched (Ci-Côjalkyl group, a linear or branched (C_rC₆)alkoxy group, a -O-(CHRi₆-CHR|₇-O)q-R^? group, a -O-P(O)(OR’)2 group, a -O-P(O)(O'M⁺)2 group, a -O-C(O)-NR|₈Ri9 group, a di(C|-C6)alkylamino(C|-C6)alkoxy group, a halogen atom, or an aldohexose of formula:

in which each R’ is independent;

it being understood that:

R’ represents a hydrogen atom or a linear or branched (C|-C₆)alkyl group,

- R|₆ represents a hydrogen atom or a (C|-C6)alkoxy(C|-C₆)alkyl group,

R_!7 represents a hydrogen atom or a hydroxy(Ci-C₆)alkyl group,

Rts represents a hydrogen atom or a (Ci-CôjalkoxyfCi-Côjalkyl group,

R[9 represents a (Ci-CtdalkoxyfCi-Côjalkyl group, a -(CHîX-NRgR./ group or a -(CH₂)_r-O-(CHR,₆-CHR_i7-O)_irR’ group, q is an integer equal to l, 2 or 3 and r is an integer equal to 0 or l, M represents a pharmaceutically acceptable monovalent cation,

The aldohexose according to the invention is preferably D-mannose. Advantageously, R_l5represents a methoxy group, a 2-methoxyethoxy group or fluorine. Preferably, the group -(CH₂)_P-Ri5 is located at ortho position of the phenyl group.

ORIGINAL

- 15 Among the preferred compounds of the invention there may be mentioned:

- JV-[5- {3-chloro-2-methyl-4-[2-(4-methylpiperazin-1 -yl)ethoxy]phenyl} -6-(4-tluoro phenyl)thieno[2,3-t/]pyrimidin-4-yl]-2-[(l-methyl-l/7-pyrazol-5-yl)methoxy]-i)phenylalanine

- A-[5- {3-chloro-2-methyl-4-[2-(4-methylpiperazin-1 -yl)ethoxy] phenyl} -6-(4-fluoro phenyl)thieno[2,3-i/]pyrimidin-4-yl]-2-[(2-ethoxypyrimidin-4-yl)methoxy]-Dphenylalanine

- A-[5-{3-chloro-2-methyI-4-[2-(4-methylpiperazin-l-yl)ethoxy]phenyl}-6-(4-fluoro phenyl )thieno[2,3-i7]pyrimidin-4-yl]-2-{[2-(2-methoxyphenyl)pyrimidin-4-yl] methoxy}-//-phenylalanine

- A-[5-{3-chloro-2-methyl-4-[2-(4-methylpiperazin-l-yl)ethoxy]phenyl}-6-(furan-2yl)thieno[2,3-i/]pyrimidin-4-yl]-2-methoxy-L>-phenylalanine iV-[5-{3-chloro-2-methyl-4-[2-(4-methylpiperazin-l-yl)ethoxy]phenyl}-6-(5fluorofuran-2-yl)thieno[2,3-i/]pyrimidin-4-yl]-2-niethoxy-Z)-phenylalanine

- A-[5-{3-chloro-2-methyl-4-[2-(4-methylpiperazin-l-yl)ethoxy]phenyl}-6-(5-fluoro furan-2-yl)thieno[2,3-i/]pyrimidin-4-yl]-2-(2,2,2-trifluoroethoxy)-£)-phenyIalanine

- .V-[5-{3-chloro-2-methyl-4-[2-(4-methylpiperazin-l-y!)ethoxy]phenyl}-6-(5-fluoro furan-2-yl)thieno[2,3-i/]pyrirnidin-4-yl]-2-(pyridin-2-ylmethoxy)-jD-phenylalanine

- ïV-l5-{3-chloro-2-methyl-4-[2-(4-methylpiperazin-l-yl)ethoxy]phenyl}-6-(5-fluoro furan-2-yl )thieno[2,3-i/]pyrimidin-4-yl]-2-[( l -methyl-1 //-pyrazol-5-yl)methoxy]D-phenylalanine /V-[5-{3-chloro-2-methyl-4-[2-(4-methylpiperazin-l-yl)ethoxy]phenyl}-6-(5-iluoro fuian-2-yl)thieno[2,3-c/]pyrimidin-4-yl]-2-[( l-ethyl-l/-/-pyrazol-5-yl)methoxy]-£)phenylalanine

- A-[5-{3-chloiO-2-methyl-4-[2-(4-methylpiperazin-l-yl)ethoxy]phenyl)-6-(5-fluoiO furan-2-yl)thieno[2,3-i/]pyrimidin-4-yl]-2-[(2-ethoxypyriniidin-4-yl)methoxy]-£>phenylalanine

- 2-[(l-butyl-l/Y-pyrazol-5-yl)methoxy]-A-[5-{3-chloro-2-methyl-4-[2-(4-methyl piperazin-1 -yl )ethoxy]phenyl} -6-(5-fluorofuran-2-yl)lhieno[2,3-i/]pyrimidin-4-yl]D-phenylalanine

ORIGINAL

- A45-{3-chloro-2-methyl-4-[2-(4-methylpiperazin-l-yl)ethoxy]phenyl}-6-(5-fluoro furan-2-yt)thieno[2,3-</lpyrimidin-4-yI]-2-{[2-(2,2,2-trifluoroethoxy)pyrimidin-4yl]methoxy}-Z)-phenylaIanine

- A-[5-{3-chloro-2-methy]-4-[2-(4-methylpiperazin-l-yl)ethoxy]phenyI}-6-(5-fluoro furan-2-yl)thieno[2,3-i/]pyrimidin-4-yl]-2-{[2-(2-methoxyphenyl)pyrimidin-4-yl] methoxy} -£)-phenylalanine

- A-[5-{3-chloro-2-methyl-4-[2-(4-methylpiperazin-l-yl)ethoxy]phenyl}-6-(prop-lyn-l-yI)thieno[2,3-i/]pyrirnidin-4-yl]-2-methoxy-£)-phenylalanine

- 2-[(l-tert-butyl-l/7-pyrazol-5-yl)methoxy]-A-[5-{3-chloro-2-methyl-4-[2-(4-methyi piperazin-1 -yl)ethoxy]phenyl} -6-(prop-1 -yn-1 -yl)thieno[2,3-i7]pyrimidin-4-yl]-Dphenylalanine

- jV-[5-{ 3-chloro-2-methyl-4-[2-(4-methy Ipiperazin-1 -yl)ethoxy]phenyl} -6-(prop-1 yn-1 -yl)thieno[2,3-rf]pyrimidin-4-yl]-2- {[2-(2-methoxyethyl)pyrimidin-4-yl] methoxy }-£)-phenylalanine

- A-[5- {3-chloro-2-methyl-4-[2-(4-methylpiperazin-1 -yl)ethoxy]phenyl} -6-(prop-1 yn-1 -yl)thieno[2,3-i/]pyrimidin-4-yl]-2- {[ l -(2,2,2-trifluoroethyl)-1 //-pyrazol-5-yl] methoxy} -£>-phenyl alanine

- A^L[5-{3-chloro-2-methyl-4-[2-(4-methylpiperazin-l-yl)ethoxy]phenyl}-6-(prop-lyn-l-yl)thieno[2,3-i/Jpyrimidin-4-y!]-2-{[2-(morpholin-4-yl)pyrimidin-4-yl] methoxy} -/)-pheny I a I ani ne

- A-[5-{3-chloiO-2-methyl-4-[2-(4-methylpiperazin-l-yl)ethoxy]phenyI}-6-(prop-lyn-l-yl)thieno[2,3-i/]pyrimidin-4-yl]-2-[[2-(2,2.2-trifluoroethoxy)pyrimidin-4-yl] methoxy} -D-phenylalanîne

- A-[5- {3-chloro-2-methy l-4-[2-(4-methylpiperazin-1 -yl)ethoxy]phenyl} -6-(prop-1 yn-l-yl)thieno[2.3-i/]pyrimidin-4-yl]-2-{[2-(2-methoxyphenyl)pyrimidin-4-yl] methoxy}-£>-phenylalanine

- 7V-[5- {3-chloro-4-[2-(dimethylamino)ethoxy]-2-methylphenyl} -6-(prop-1 -yn-1 -yl ) thieno[2,3-t/]pyrimidin-4-yl]-2-{[l-(2,2,2-trifluoroethyl)-l//-pyrazol-5-yl] methoxy J-Û-phenylalanine

- /V-[5- {3-chloro-4-[2-(dimethylamino)ethoxy]-2-methylpheny l }-6-(prop-1 -yn-1 -yl) thieno[2,3-rfJpyrimidin-4-yl]-2-{[2-(morpholin-4-yl)pyrimidin-4-yl]methoxy}-£>phenylalanine _<

ORIGINAL

- I7-

- Α-[5- {3-chloro-4-[2-(dimethylamino)ethoxy]-2-methylphenyl }-6-(prop-1 -yn-1 -yl) thieno[2,3-i/]pyrimidin-4-yi]-2-{[2-(2,2,2-tritluoroethoxy)pyrimidin-4-yl] methoxy} -D-phenylalanine

- A-[5- {3-chloro-4-[2-(dimethylamino)ethoxy]-2-methyIphenyl} -6-(prop-1 -yn-1 -yl) thieno[2,3-i/]pyrimidin-4-yl]-2-{[2-(2-methoxyphenyl)pyrimidin-4-yl]methoxy}-£>phenylalanine

- A-[5-{3-chloro-4-[2-(dimethylamino)ethoxy]-2-methylphenyl}-6-(4-fluorophenyl) thieno[2,3-t/Jpyrimidin-4-yl]-2-({2-[2-(2-methoxyethoxy)phenyl]pyrimidin-4-yl} methoxy)-D-phenylalanine

- ethyl /7-((55^)-5- {3-chloro-2-methyl-4-[2-(4-methylpiperazin-1 -yl)ethoxy]phenyl} -

6-(4-fluorophenyl)thieno[2,3-i/]pyrimidm-4-yl]-2-{[2-(2-methoxyphenyl) pyrimidin-4-yl]methoxy}-Z)-phenylalanmate ethyl A-[(5S_a)-5- {3-chloro-2-methyl-4-[2-(4-methylpiperazin-l -yl)ethoxy]phenyl} 6-(prop-1 -yn-1 -y l)thieno [2,3</]pyrîmidin-4-yl]-2- {[2-(2-methoxyphenyl ) pyrimidin-4-yl]methoxy}-jD-phenylalaninate

- ethyl A-[(55_a)-5-{3-chloro-4-[2-(dimethylamino)ethoxy]-2-methylphenyl}-6-(propl-yn-l-yl)thieno[2,3-i:/]pyrimidin-4-yl]-2-{[2-(2-methoxyphenyl)pyrimidin-4-yl] methoxy }-Z>-phenylalaninate A-[5-{3,5-dichloro-2,6-dimethyl-4-[2-(4-methylpiperazin-l-yl) ethoxy]phenyl}-6-(4-fluorophenyl)thieno[2,3-i/]pyrimidin-4-yl]-2-{[2-(2niethoxyphenyI)pyrimidin-4-yl]methoxy}-D-phenylalanine

The invention relates also to a process for the préparation of compounds of formula (I), which process is characterised in that there is used as starting material the compound of formula (Il-a):

ci^^A^z ^(,l'^a) wherein Z represents bromine or iodine and A is as defined for formula (l) in which 1 is linked to the chlorine atom and 2 is linked to the Z group, which compound of formula (ΙΙ-a) is subjected to coupling with a compound of

ORIGINAL

- I8formula (III):

Alk

R]₂ (III) wherein R_f„ R|₂, E and n are as defined for formula (I), and Alk represents a linear or branched (Cj-C₆)alkyl group, to yield the compound of formula (IV):

Alk

^RI2 wherein R_f), R_l2, A. E and n are as defined for formula (I) and, Z and Alk is as defined before.

compound of formula (IV) which is further subjected to coupling with compound of formula (V):

(V) wherein R|, R₂, R5, X and Y are as defined for formula (I), and R_B! and R_B2 represent a hydrogen atom, a linear or branched (Ci-C₆) alkyl group, or R_Bi and R_B2 form with the oxygen carrying them an optionally methylated ring,

ORIGINAL

- 19to yield the compound of formula (VI):

Alk

(VI) wherein R], R₂, R₅, R₆, R_l2, X, Y. A, E and n are as defined for formula (I) and Alk is as defined before.

the Alk-O-C(O)- ester function of which compound of formula (VI) is hydrolysed to yield the carboxylic acid, which may optionally be reacted with an alcohol of formula R₇’-OH or a chlorinated compound of formula R₇’-CI wherein R₇’ represents a linear or branched (C|-C₈)alkyl group, a -CHR^Rt, group, an aryl group, a heteroaryl group, an arylalkyfiCi-Cé) group, or a heteroarylalkyl(C_rC₆) group, R_a and R_b are as defined for formula (l), to yield the compound of formula (I), which may be purified according to a conventional séparation technique, which is converted, if desired, into its addition salts with a pharmaceutically acceptable acid or base and which is optionally separated into its isomers according to a conventional séparation technique, it being understood that at any moment considered appropriate during the course of the process described above, some groups (hydroxy, amino...) of the starting reagents or of the synthesis intermediates can be protected, subsequently deprotected and funciionalized, as required by the synthesis.

In an other embodiment of the invention, compounds of formula (I) may be obtained using an alternative process, which process is characterised in that there is used as starting material the compound of formula (Il-b):

ORIGINAL c/S (Π-b) wherein A is as defined in formula (I) in which 1 is linked to the chlorine atom and 2 is linked to the iodine atom.

which compound of formula (Π-b) is subjected to coupling with a compound of formula (V):

(V) wherein R], R₂, R5, X and Y are as defined for formula (I), and R_B| and R_B2 represent a hydrogen atom, a linear or branched (C_rC₆) alkyl group, or R_Bi and R_B2 form with the oxygen carrying them an optionally methylated ring, !0 to yield the compound of formula (Vil):

(Vil) wherein R], R₂, R5, A, X and Y are as defined in formula (I), which compound of formula (VII) is further subjected to coupling with a compound of formula (III):

ORIGINAL

Alk

*12 (ΙΠ) wherein R₆, R_f2, E and n are as defined for formula (I), and Alk represents a linear or branched (Ci-Cô)alkyl group, to yield the compound of formula (VI):

wherein R_h R₂, R?, R<,, R,?, X, Y, A, E and n are as defined for formula (I) and Alk is as defined before, the Alk-O-C(O)- ester function of which compound of formula (VI) is hydrolysed to yield the carboxylic acid, which may optional!y be reacted with an alcohol of formula R₇'-0H or 10 a chlorinated compound of formula R₇’-Cl wherein R₇’ represents a linear or branched (Ci-Cgjalkyl group, a -CHR_aRf, group, an aryl group, a heteroaryl group, an arylalkyl(C|-C₀) group, or a heteroarylalkyl(C_rC₆) group, R_a and R_b are as defined for formula (I), to yield the compound of formula (I), which may be purified according to a conventional I5 séparation technique, which is converted, if desired. into its addition salts with a pharmaceutically acceptable acid or base and which is optionally separated into its isomers according to a conventional séparation technique,

ORIGINAL

-22 it being understood that at any moment considered appropriate during the course of the process described above, some groups (hydroxy, amino...) of the starting reagents or ofthe synthesis intennediates can be protected, subsequently deprotected and functionalized, as required by the synthesis.

The compounds of formulae (Π-a), (ΙΙ-b), (III), (V), R₇’-OH and R₇’-Cl are either commercially available or can be obtained by the person skilled in the art using conventional Chemical reactions described in the literature.

Pharmacological study of the compounds of the invention has shown that they hâve proapoptotic properties. The ability to reactivate the apoptotic process in cancerous cells is of major therapeutic interest in the treatment of cancers and of immune and auto-immune diseases.

More especially, the compounds according to the invention will be useful in the treatment of chemo- or radio-resistant cancers.

Among the cancer treatments envisaged there may be mentioned, without implying any limitation, treatment of cancers of the bladder, brain, breast and utérus, chronic lymphoid leukaemias, cancer of the colon, œsophagus and liver, lymphoblastic leukaemias, acute myeloid leukaemias, lymphomas, melanomas, malignant haemopathies, myelomas, ovarian cancer, non-small-cell lung cancer, prostate cancer, pancreatic cancer and small-cell lung cancer.

The present invention relates also to pharmaceutical compositions comprising at least one compound of formula (I) in combination with one or more pharmaceutically acceptable excipients.

Among the pharmaceutical compositions according to the invention there may be mentioned more especially those that are suitable for oral, parentéral, nasal, per- or trans-cutaneous, rectal, perlingual, ocular or respiratory administration, especially tablets

ORIGINAL

-23or dragées, sublingual tablets, sachets, paquets, capsules, glossettes, lozenges, suppositories, creams, ointments, dermal gels, and drinkable or injectable ampoules.

The dosage varies according to the sex, âge and weight of the patient, the administration route, the nature of the therapeutic indication, or of any associated treatments, and ranges from 0.01 mg to l g per 24 hours in one or more administrations.

Furthermore, the présent invention relates also to the combination of a compound of formula (I) with an anticancer agent selected from genotoxic agents, mitotic poisons, antimetabolites, proteasome inhibitors, kinase inhibitors and antibodies, and also to pharmaceutical compositions comprising that type of combination and their use in the manufacture of médicaments for use in the treatment of cancer.

Advantageously, the présent invention relates to the combination of a compound ot formula (Γ) with an EGFR inhibitor, and also to pharmaceutical compositions comprising that type of combination.

In another embodiment, the présent invention relates to the combination of a compound of formula (l) with a mT0R/PI3K inhibitor, and also to pharmaceutical compositions comprising that type of combination.

In a preferred embodiment, the présent invention relates to the combination of a compound of formula (I) with a MEK inhibitor, and also to pharmaceutical compositions comprising that type of combination.

Preferably, the présent invention relates to the combination of a compound of formula (I) with a HER2 inhibitor, and also to pharmaceutical compositions comprising that type of combination.

Advantageously, the présent invention relates to the combination of a compound of formula (I) with a RAF inhibitor, and also to pharmaceutical compositions comprising that type of combination.

In another embodiment, the présent invention relates to the combination of a compound of formula (I) with a EGFR/IIER2 inhibitor, and also to pharmaceutical compositions comprising that type of combination.

ORIGINAL

-24In a preferred embodiment. the présent invention relates to the combination of a compound of formula (I) with a taxane, and also to pharmaceutical compositions comprising that type of combination.

In another embodiment, the présent invention relates to the combination of a compound of formula (I) with a protéasome inhibitor, an immunomodulator or an alkylating agent, and also to pharmaceutical compositions comprising that type of combination.

The combination of a compound of formula (I) with an anticancer agent may be administered simultaneously or sequentially. The administration route is preferably the oral route, and the corresponding pharmaceutical compositions may allow the instantaneous or delayed release of the active ingrédients. The compounds of the combination may moreover be administered in the form of two separate pharmaceutical compositions, each containing one of the active ingrédients, or in the form of a single pharmaceutical composition, in which the active ingrédients are in admixture.

The compounds of the invention may also be used in combination with radiotherapy in the treatment of cancer.

Finally, the compounds of the invention may be linked to monoclonal antibodies or fragments thereof or linked to scaffold proteins that can be related or not to monoclonal antibodies.

Antibody fragments must be understood as fragments of Fv, scFv, Fab, F(ab‘)2, F(ab'), scFv-Fc type or diabodies, which generally hâve the same specificity of binding as the antibody from which they are descended. According to the présent invention, antibody fragments of the invention can be obtained starting from antibodies by methods such as digestion by enzymes, such as pepsin or papain, and/or by cleavage of the disulfide bridges by Chemical réduction. In another manner, the antibody fragments comprised in the présent invention can be obtained by techniques of genetîc recombination likewise well known to the person skilled in the art or else by peptide synthesis by means of, for example, automatic peptide synthesizers such as those supplied by the company Applied Biosystems, etc.

ORIGINAL

-25Scaffold proteins that can be related or not to monoclonal antibodies are understood to mean a protein that contains or not an immunoglobulin fold and that yields a binding capacity similar to a monoclonal antibody. The man skilled in the art knows how to select the protein scaffold. More partîcularly, it is known that, to be selected, such a scaffold should display several features as follow (Skerra A., J. Mol, Recogn. 2000, B, 167-187): phylogenetically good conservation, robust architecture with a well-known threedimensionai molecular organization (such as, for example, crystallography or NMR), small size, no or only a low degree of post-translational modifications, easy to produce, express and purify. Such a protein scaffold can be, but without limitation, a structure selected from the group consisting in fibronectin and preferentially the tenth fibronectin type III domain (FNfnlO), lipocalin, anticalin (Skerra A., J. Biotechnol. 2001, 74(4):257-75), the protein Z dérivative from the domain B of staphylococcal protein A, thioredoxin A or any protein with a repeated domain such as an ankyrin repeat (Kohl et ai, PNAS 2003, 100(4). 1700-1705), armadillo repeat, leucine-rich repeat or tetratricopeptide repeat. There could also be mentioned a scaffold dérivative from toxins (such as, for example, scorpion, insect, plant or mollusc toxins) or protein inhibitors of neuronal nitric oxide synthase (PIN).

The following Préparations and Examples illustrate the invention but do not limit it in any way.

General Procedures

Ail reagents obtained from commercial sources were used without further purification. Anhydrous solvents were obtained from commercial sources and used without further drying.

Flash chromatography was performed on ISCO CombiFlash Rf 200i with pre-packed silica-gel cartridges (Redi&p®A_r Gold High Performance).

Thin layer chromatography was conducted with 5x10 cm plates coated with Merck Type 60 F254 silica-gel.

ORIGINAL

-26Microwave heating was performed in an Anton Pair MonoWave or CEM Discover® instrument.

Préparative HPLC purifications were performed on an Armen Spot Liquid Chromatography System with a Gemini-NX’ ΙΟ μΜ Cl8, 250 mm χ 50 mm i.d. column running at a flow rate of 118 mL min’¹ with UV diode array détection (210 - 400 nm) using 25 mM aqueous NH4HCO3 solution and MeCN as eluents uniess specified otherwise.

Analytical LC-MS: The compounds of the present invention were characterized by high performance liquid chromatography-mass spectroscopy (HPLC-MS) on Agilent HP 1200 with Agilent 6140 quadrupole LC/MS, operating in positive or négative ion electrospray ionisation mode. Molecular weight scan range is 100 to 1350. Parallel UV détection was done at 210 nm and 254 nm. Sampies were supplied as a 1 mM solution in ACN, or in THF/H2O (1:1) with 5 pL loop injection. LCMS analyses were performed on two instruments, one of which was operated with basic, and the other with acidic eluents.

Basic LCMS: Gemini-NX, 3 pm, Cl8, 50 mm x 3.00 mm i.d, column at 23 °C, at a flow rate of 1 mL min'¹ using 5 mM ammonium bicarbonate (Solvent A) and acetonitrile (Solvent B) with a gradient starting from 100% Solvent A and finishing at 100% Solvent B over various/certain duration of lime.

Acidic LCMS: ZORBAX Eclipse XDB-C18, 1.8 pm, 50 mm x 4.6 mm i.d. column at 40 °C, at a flow rate of 1 mL min’¹ using 0.02% v/v aqueous formic acid (Solvent A) and 0.02% v/v formic acid in acetonitrile (Solvent B) with a gradient starting from 100% Solvent A and finishing at 100% Solvent B over various/certain duration of time.

‘H-NMR measurements were performed on Broker Avance 111 500 MHz spectrometer and Broker Avance III 400 MHz spectrometer, using DMSO-d₆ or CDCl·; as solvent. ’H NMR data ts in the form of delta values, given in part per million (ppm), using the residual peak of the solvent (2.50 ppm for DMSO-d₆ and 7.26 ppm for CDCI3) as internai standard. Splitting patterns are designated as: s (singlet), d (doublet), t (triplet), q (quartet), quint

ORIGINAL

-27(quintet), m (multiplet), br s (broad singlet), dd (doublet of doublets), td (triplet of doublets), dt (doublet of triplets), ddd (doublet of doublet of doublets).

Combination gas chromatography and low resolution mass spectrometry were performed on Agilent 6850 gas chromatograph and Agilent 5975C mass spectrometer using 15 m χ 5 0.25 mm column with 0.25 pm HP-5MS coating and hélium as carrier gas. Ion source: ΕΓ, eV, 230°C, quadrupole: l50°C, interface: 300°C.

HRMS were determined on a Shimadzu IT-TOF, ion source température 200°C, ESI +/-, ionization voltage: (+-)4.5 kV. Mass resolution min. 10000.

Elementary analyses were performed on a Thermo Flash EA 1112 Elemental Analyzer.

10	List of abbreviations
	Abbreviation Name 2-Me-THF 2-methyl-tetrahydrofurane Ac acetyl Ad adamantyl
I5	aq. aqueous AtaPhos bis(di-re>7-butyl(4-dimethylaminophenyl)phosphine) dichloropalladium(II) BuPAd2 butyl-di(adamant-1 -y l)phosphane ce. concentrated
20	DAST diethyiaminosulfur trifluoride dba dibenzylideneacetone DCM methylene chloride DIPA diisopropy lamine DMA dimethylacetamide
25	DME l ,2-dimethoxyethane DMF dimethylformamide

ORIGINAL

	-28-
DMSO	dimethy] sulfoxide
dppf	1,1 '-bis(diphenylphosphino)ferrocene
eq. Et	équivalent ethyl
HILIC	hydrophilic interaction liquid chromatography
HMDS	hexamethyldisilazane
'Pr	isopropyl
LDA	lithium diisopropylamide
Me	methyl
MeCN	acetonitrile
MTBE	methyl Ze/7-butyl ether
MW	micro wave
NBS	jV-bromosuccinimide
ⁿBu	rt-butyl
NCS	/V-chlorosuccinimide
Ph	phenyl
P'Buj x HBF₄	tri-re?7-butylphosphonium tetrafluoroborate
PCy₃ x 11BF₄	tricyclohexylphosphonium tetrafluoroborate
Q-Phos r.t.	1,2,3,4,5-pentaphenyI-l ’-(di-/e/7-butylphosphino)ferrocene room température
TBAF	tetrabutyl ammonium fluoride
'Bu	te/7-butyl
TEA	triethylamine
TFA	trifluoroacetic acid
THF	tetrahydrofurane
TIPSC1	trîisopropylsilyl chloride
XantPhos	4,5-bis(diphenyiphosphino)-9,9-dimethylxanthene

General Procedure la:

l eq. of the appropriate 4-chloro-thieno[2.3-J]pyrimidine dérivative, 2 eq. of the appropriate amino acid dérivative and 2 eq. K₂CO₃ were mixed in 'BuOH: water 4:1

ORIGINAL

-29(4 mL/mmol) and stirred at reflux température (or in MW reactor at 100 °C if it is needed) until no further conversion was observed. The mixture was then diluted with water, acidified with IM HCl solution (to pH = l, or to pH = 6 in the presence of a basic amino group) and extracted with EtOAc, or the precipitate formed after acidification was isolated by filtration. In the case of extractive purification the combined organic phases were washed with brine, dried over MgSÜ4, filtered and the filtrate was concentrated under reduced pressure. The crude product was purified via préparative reversed phase chromatography using 25 mM aqueous NH4HCO3 and acetonitrile as eluents unless otherwise stated.

General Procedure Ib:

l eq. of the appropriate 4-chloro-thieno[2,3-i/]pyrimidine dérivative, 2 eq. of the appropriate amino acid dérivative and 3 eq. K2CO3 were mixed in DMSO (IO mL/mmol) and stirred at 50 °C until no further conversion was observed. The mixture was then diluted with water, acidified with IM HCl solution (to pH = 1, or to pH = 6 in the presence of a basic amino group) and extracted with EtOAc, or the precipitate formed after acidification was isolated by filtration. In the case of extractive purification the combined organic phases were washed with brine, dried over MgSO₄, filtered and the filtrate was concentrated under reduced pressure. The crude product was purified via préparative reversed phase chromatography using 25 mM aqueous NH4HCO3 and acetonitrile as eluents unless otherwise stated.

General Procedure le:

eq. of the appropriate 4-chloro-thieno[2,3-i/]pyrimidine dérivative, 1.5 eq. of the appropriate amino acid dérivative and 1.5 eq. CS2CO3 were mixed in DMSO (6 mL/mmol) and stirred at 70 °C until no further conversion was observed. The mixture was then diluted with water, acidified with IM HCl solution (to pH = 1, or to pH = 6 in the presence of a basic amino group) and extracted with EtOAc, or the precipitate formed after acidification was isolated by filtration. In the case of extractive purification the combined organic phases were washed with brine, dried over MgSO₄, filtered and the filtrate was concentrated under reduced pressure. The crude product was purified via préparative

ORIGINAL

-30reversed phase chromatography using 25 mM aqueous NH4HCO3 solution and acetonitrile as eluents unless otherwise stated.

General Procedure lia:

l eq. of the appropriate 5- (or 6)-iodo-thieno[2,3-i/]pyrimidine dérivative and 3 eq. of the appropriate boronic acid dérivative were dissolved in DME (15 mL/mmol), then 5 eq. K₂CO₃, 0.2 eq. Pd₂dba₃, 0.4 eq. BuPAd₂ and water (5 mL/mmol) were added and the mixture was stirred at 60 °C in MW reactor until no further conversion was observed. The volatiles were then removed in vacuo and the residue was purified via préparative reversed phase chromatography, using 25 mM aqueous NH4HCO3 solution and acetonitrile as eluents unless otherwise stated.

General Procedure Ilb:

l eq. of the appropriate 5-iodo-thieno[2,3-J]pyrimidine dérivative and 5 eq. of the appropriate boronic acid dérivative were dissolved in 2-Me-THF (8 mL/mmol), then 5 eq. K2CO3, O.l eq. Q-Phos and 0.05 eq. Pd₂dba₃ were added and the mixture was stirred at 80 °C until no further conversion was observed. The mixture was filtered through a pad of Celite, the filtrate was concentrated under reduced pressure. The crude product was purified via flash chromatography using heptane and EtOAc as eluents unless otherwise stated.

General Procedure Ile:

l eq. of the appropriate 5-(or 6)-iodo-thieno[2,3-i7]pyrimidine dérivative and l .1 eq. of the appropriate boronic acid dérivative were dissolved in 2-Me-THF (8 mL/mmol), then

l.l eq. Ag₂CO₃ and O.l eq. Pd(PPh₃)₄ were added and the mixture was stirred at 100 °C until no further conversion was observed. The mixture was filtered through a pad of Celite, the filtrate was concentrated under reduced pressure. The crude product was purified via flash chromatography using heptane and EtOAc as eluents unless otherwise stated.

General Procedure Ild:

l eq. of the appropriate 5-iodo-thieno[2,3-i/]pyrimidine dérivative and 3 eq. of the appropriate boronic acid dérivative were dissolved in dioxaneiwater 2:l mixture

ORIGINAL

-3l (ΙΟ mL/mmol), then 2 eq. Cs₂CO₃, 5 mol% Pd(OAc)₂ and 0.2 eq. P'Bu₃xHBF₄ were added and the mixture was stirred at 120 °C in MW reactor until no further conversion was observed. The mixture was neutralized with IM HCl solution and extracted with DCM. The combined organic phases were dried over Na₂SO₄, fîitered and the filtrate was concentrated in vacuo. The crude product was purified via préparative reversed phase chromatography using O.l % aqueous TFA solution and acetonitrile as eluents unless otherwise stated.

General Procedure Ilia:

l eq. of the appropriate 6-iodo-thieno[2,3-i/]pyrimidine derivative and 4 eq. of the appropriate boronîc acid derivative were dissolved in dioxane:water 4:l mixture ( 10 mL/mmol), then 2.2 eq. Cs₂CO₃ and O.l eq. Pd(dppf)Cl₂ were added and the mixture was stirred at 40°C until no further conversion was observed. The mixture was then diluted with water and extracted with DCM. The combined organic phases were washed with water, dried over MgSO₄, fîitered and the filtrate was concentrated under reduced pressure. The crude product was purified using préparative reversed phase chromatography using 25 mM aqueous NH₄HCO₃ solution and acetonitrile as eluents unless otherwise stated.

General Procedure 111b:

l eq. of the appropriate 6-iodo-thieno[2,3-i7]pyrimidine derivative and 3 eq. of the appropriate boronic acid derivative were dissolved in THF: water l:l mixture (10 mL/mmol), then 3 eq. Cs₂CO₃ and O.l eq. AtaPhos were added and the mixture was stirred at 100 °C in MW reactor until no further conversion was observed. The volatiles were evaporated under reduced pressure, and the residue was purified via préparative reversed phase chromatography using 25 mM aqueous NH₄HCO₃ solution and acetonitrile as eluents unless otherwise stated.

General Procedure IVa:

l eq. Préparation 4i was dissolved in dry THF (5 mL/mmol) and cooled to -78 °C. LDA solution (1.2 eq. 2M in THF, heptane, EtPh) was added dropwise under Argon and the mixture was stirred for 1.5 hours. Then 1.2 eq. of the appropriate electrophilic-reagent either in solution (dissolved in 3mL/mmol dry THF), or neat was added at -78 °C and the

ORIGINAL

-32mixture was allowed to warm up to r.t. It was stirred until no further conversion was observed. The reaction mixture was quenched by the careful addition of cc. NH₄Cl solution. The mixture was extracted with MTBE, the organic layer was washed with brine, dried over Na₂SO₄, filtered and the filtrate was concentrated under reduced pressure. The crude product was purified via flash chromatography using heptane and EtOAc as eluents unless otherwise stated.

General procedure Va:

eq. of the appropriate acetal was stirred in 2M HCl solution (3 mL/mniol) at 60 °C until no further conversion was observed. The reaction mixture was cooled to 0 °C, then 5.7 eq. NaOH was added portionwise. The pH was adjusted to 8 using 10% K2CO3 solution, then eq. sodium borohydride was added portionwise keeping the température below 5 °C. After the addition the mixture was stirred at 0 °C until no further conversion was observed. The mixture was extracted with EtOAc, the combined organic phases were dried over Na₂SO4, filtered and the filtrate was concentrated under reduced pressure. The crude product was purified via flash chromatography using heptane and EtOAc as eluents unless otherwise stated.

General Procedure Vb:

Step A

To a solution of l eq. ofthe appropriate Λ'-alkyl pyrazole in dry THF (1.5 mL/mmol),

l.l eq. BuLi was added dropwise at -78 °C. The mixture was stirred for 30 minutes and then allowed to warm up to 0 °C where it was stirred for 30 minutes, then cooled back to -78 °C again. I.l eq. DMF was added dropwise, then the reaction mixture was allowed to reach r.t. and it was stirred ovemight. The mixture was quenched with cc. NH₄Cl solution. The phases were separated and the aqueous layer was extracted with EtOAc. The combined organic layers were dried over Na₂SO₄, filtered and the filtrate was concentrated under reduced pressure. The residue was used in the next step without further purification.

Step B

ORIGINAL

To a solution of l eq. of the appropriate crude aldéhyde in EtOH (0.5 mL/mmol), 1.3 eq. sodium borohydride was added portionwise at -15 °C and the reaction mixture was stirred at r.t. until no further conversion was observed. The mixture was poured onto crushed ice and stirred for 16 hours. The precipitate was filtered off, and the filtrate was concentrated under reduced pressure. The oily phase was separated, and the aqueous layer was extracted with EtOAc. The combined organic layers were dried over Na₂SO₄, filtered and the filtrate was concentrated under reduced pressure. The product was further purified by flash chromatography if necessary.

General procedure Vc:

To the mixture of l .2 eq. of the appropriate amidine sait and l eq. Préparation 8a in dry methanol (0.5 mL/mmol) l .2 eq. sodium methoxide was added portionwise and the mixture was stirred at 75 °C until no further conversion was observed. The reaction mixture was cooled and concentrated under reduced pressure. Water was added to the residue, and it was extracted with DCM. The combined organic layers were dried over MgSO₄, filtered and the filtrate was concentrated under reduced pressure. The crude product was purified via flash chromatography using heptane and EtOAc as eluents unless otherwise stated.

General Procedure Vd:

To the mixture of l .2 eq. of the appropriate hydrazine or hydrazine hydrochloride and l eq. Préparation 8a in dry methanol (0.5 mL/mmol) 1.2 eq. sodium methoxide was added portionwise and the mixture was stirred at 75 °C until no further conversion was observed. The reaction mixture was cooled and concentrated under reduced pressure. Water was added to the residue, and it was extracted with DCM. The combined organic phases were dried over MgSO₄, filtered and the filtrate was concentrated under reduced pressure. The crude product was purified via flash chromatography using heptane and EtOAc as eluents unless otherwise stated.

General procedure Ve:

l eq. of the appropriate acetal was stirred with IM HCl solution (3 mL/mmol) at 50 °C until no further conversion was observed. The reaction mixture was cooled to 0 °C, then

ORIGINAL

2.85 eq. solid NaOH was added portionwise. The pH was adjusted to 8 using 10 % K₂CO₃solution, then 2 eq. sodium borohydride was added portionwise keeping the température below 5 °C and stirred at 0 °C until no further conversion was observed. The mixture was extracted with EtOAc, the combined organic phases were dried over Na₂SO₄, filtered and 5 the filtrate was concentrated under reduced pressure. The crude product was purified via flash chromatography using heptane and EtOAc as eluents unless otherwise stated.

General Procedure VI:

l eq. of the appropriate phénol dérivative, 2 eq. of the appropriate alcohol dérivative, and eq. PPh₃ were dissolved in dry toluene (7 mL/mmol) under N₂ atmosphère, then 3 eq.

di-/er/-butyl azodicarboxylate was added at r.t. Then the mixture was stirred at 50 °C until no further conversion was observed. The volatiles were removed in vacuo and the residue was purified via flash chromatography using heptane and EtOAc (and MeOH if needed) as eluents. If necessary', the product was further purified via préparative reversed phase chromatography using 25 mM aqueous NH4HCO3 solution and acetonitrile as eluents 15 unless otherwise stated.

General Procedure VII:

l eq. of the appropriate ester dérivative was dissolved in THF (15 mL/mmol) then IO eq. LiOH*H₂O and water (15 mL/mmol) were added. The mixture was stirred at r.t. (or at 60 °C if needed) until no further conversion was observed. The pH was adjusted to 6 with 20 IM HCl solution, then the mixture was diluted with brine, extracted with DCM or EtOAc.

The organic layer was dried over Na₂SO₄, filtered and the filtrate was concentrated under reduced pressure. The crude product was purified via préparative reversed phase chromatography using 25 mM aqueous NH4HCO3 solution and acetonitrile as eluents unless otherwise stated.

General Procedure VIII:

l eq. of the appropriate indole dérivative and 2 eq. of the appropriate alcohol dérivative were dissolved in dry toluene (8 mL/mmol) under N₂ atmosphère, and the mixture was cooled to 0 °C, then 2 eq. 2-(tributyl-phosphanylidene)acetonitrile was added. Then the mixture was heated to 100 °C and stirred until no further conversion was observed. The

ORIGINAL

- 35 volatiles were removed in vacuo, then water (4 mL/mmol) 2M NaOH solution (l mL/mmol) were added and the mixture was stirred until no further conversion was observed. The mixture was then acidified with IM HCl solution to pH = 6 and extracted with DCM. The combined organic phases were dried over Na₂SO4, filtered and the filtrate was concentrated under reduced pressure. The crude product was purified via préparative reversed phase chromatography using aqueous 40 mM NH4OAC (pH = 4, adjusted with AcOH) solution and acetonitrile as eluents unless otherwise stated.

General procedure IXa:

Step A l eq. Préparation 9b was dissolved in dry toluene (8 mL/mmol). then LIS eq. PPh₃,

l.l eq. of the appropriate alcohol dérivative and 1.18 eq. diethylazodicarboxylate (40 % solution in toluene) were added at r.t. The mixture was stirred at r.t. until no further conversion was observed. The resulting precipitate was filtered off and the filtrate was washed sequentially with 10 % KHSO4 solution, water, sat. NaHCCh solution and water again. The organic phase was dried over Na2SO4, filtered and the filtrate was concentrated under reduced pressure. The residue was agitated with diethyl-ether (5 mL/mmol), the insoluble material was filtered off and the filtrate was concentrated under reduced pressure to obtain the crude product.

Step B

The product of Step A was treated with 10 eq. HCl solution (4.9M in MeOH) and it was stirred at r.t. until no further conversion was observed. Then the mixture was concentrated under reduced pressure. The residue was partitioned between cold EtOAc and ice-cold water, the phases were separated and the organic phase was extracted with ice-cold 5 % KHSO4 solution. The combined aqueous phase was basified with solid Na₂CO₃, and the product was extracted with EtOAc. The organic phase was dried over Na₂SO4, filtered and the filtrate was concentrated under reduced pressure to obtain the methyl ester of the title product.

Step C

ORIGINAL

- 36 l eq. of the methyl ester obtained in Step B was dissolved in MeOH (9 mL/mmol), then l .05 eq. NaOH and water (l mL/mmol) were added and the mixture was stirred at r.t. until no further conversion was observed. Methanol was removed under reduced pressure and the mixture was neutralized using l M HCl solution, then it was extracted with DCM. The organic phase was dried over Na₂SÛ4, filtered and the filtrate was concentrated under reduced pressure to obtain the O-alkylated amino acid dérivative which was used without further purification.

General procedure IXb:

Step A l eq. Préparation 9b was dissolved in dry DMF ( 10 mL/mmol) and 4 eq. IGCO3 and 2 eq. of the appropriate alkylating agent was added at r.t. The mixture was stirred at 50 °C until no further conversion was observed. The mixture was diluted with water, then extracted with DCM. The organic phase was dried over Na₂SO4, filtered and the filtrate was concentrated under reduced pressure. The crude materiai was purified via flash chromatography using DCM and methanol as eluents unless otherwise stated.

Step B and Step C are the same as described in General procedure IXa.

Préparation la: 5-Bromo-4-chloro-6-iodo-thieno[2,3-r/|pyrimidinc

Step A: 6-lodo-3H-lhieno[2,3-d]pyrimidin-4-one

A 2 L round bottomed flask equipped with mechanical stirrer, thermometer and reflux condenser was charged with the solution of 433 mL acetic acid, 13 mL sulfuric acid and 87 mL water. 69.3 g 3/7-thieno[2,3-i/]pyrimidin-4-one (0.46 mol), 51.9 g periodic acid (0.23 mol) and 104 g iodine (0.41 mol) were added to the stirred solution, which was heated lo 60 °C for l hour. The resulting suspension was cooled to r.t., filtered off, washed with a mixture of acetic acid and water (5:l) and then with diethyl ether. The resulting beige crystalline solid was air dried. ’l-l NMR (500 MHz, DMSO-d₆) δ: 12.57 (br s, IH), 8.09 (s, IH), 7.65 (s, IH)

ORIGINAL

-37Step B: 4-Chloro-6-iodo-lhieno[2,3-à]pyrimidine

A l L round bottomed flask equipped with mechanical stirrer, thermonieter, reflux condenser and a CaC^-tube was charged with H3 mL phosphorous oxychloride and 35 mL MA-dimethylaniline (0.29 mol). 75.54 g 6-iodo-3A-thieno[2,3-i/]pyrimidin-4-one (0.27 mol) was added to the mixture in portions during 5 minutes. The reaction mixture was stirred at 105 °C for l hour. The resulting suspension was cooled to 10 °C, filtered and washed with hexane. The crude product was added to ice water and stirred for 10 minutes, filtered off, washed with cold water, diethyl ether and air dried. Beige crystalline solid was obtained. 'H NMR (400 MHz, DMSO-d₆) δ; 8.89 (s, IH), 7.98 (s, IH)

Step C: Préparation la

A 2 L round bottomed flask equipped with mechanical stirrer, thermometer and a bubbler was charged with 600 mL acetonitrile. 84.9 g 4-chloro-6-iodo-thieno[2,3-i/]pyrimidine (0.29 mol), 50.9 g NBS (0.29 moi) and 8.5 mL tetrafluoroborîc acid diethyl ether complex were added. The reaction mixture was stirred at r.t. for 16 hours. Further 22.9 g (O.l2 mol) NBS was added to the mixture in three portions. After cooling the suspension to 0 °C and stirring for further l hour the precipitate was filtered off, washed with acetonitrile and air dried. The product was obtained as beige crystalline solid. ^!H NMR (500 MHz, DMSO-de) δ: 8.88 (s, IH)

Préparation 1b: 4-Chloro-5,6-diiodo-thieno[2,3-d]pyrimidine

Step A : 5,6-Diiodo-3\A-thieno[2,3-d]pyrimidin-4-one

To a well stirred slurry of 61.3 g 3A-thieno[2,3-c/]pyrimidin-4-one (396 mmol), 92.4 g periodic acid (405 mmol), l L acetic acid, 200 mL water, 6 mL cc. sulfuric acid, and 203 g iodine (799 mmol) were added. The reaction mixture was heated to 110 °C and stirred for 3 hours. The suspension was cooled to r.t. then 940 mL diethyl ether was added and the mixture was stirred further at 10 °C for 30 minutes. The precipitate was filtered off, washed with a 2:l mixture of diethyl ether and éthanol (100 mL), finally with diethyl ether (3 χ 250 mL), then it was air dried to give the product as a tan powder. ‘H NMR (500 MHz, DMSO-d₆) δ: 12.60 (br s, l H), 8.13 (s, l H)

ORIGINAL

-38Step B: Préparation 1b

To a well stirred slurry of 180 g 5,6-diiodo-3/7-thieno[2,3-i/]pyrimidin-4-one (445 mmol) in 2.5 L phosphorous oxychloride 64 mL A’jV-dimethylaniline was added. The reaction mixture was heated to 105 °C and stirred for 1.5 hours. The resulting suspension was cooled to r.t. and 1.5 L hexane was added and it was stirred for an additional 20 minutes. The precipitate was filtered off, washed with hexane (3 χ 500 mL) and water (3 * 100 mL) then air dried to give the product as a grey crystalline solid. ^]H NMR (400 MHz, DMSOd₆) Ô: 8.88 (s, IH)

Préparation le: 4-Chloro-5-iodo-thieno|2,3-i/]pyrimidine

52.8 g Préparation 1b (125 mmol) was dissolved in 400 mL dry THF and cooled to 0 °C. 100 mL ’BuMgCl (200 mmol, 2M in diethyl ether) was added over 15 minutes. Then 50 mL water was added and the solution was decanted and concentrated under reduced pressure. The crude product was sonicated in a mixture of acetonitrile and water (3:l) and then collected by filtration. ’H NMR (400 MHz, DMSO-d&) δ: 8.95 (s, IH), 8.45 (s, IH)

Préparation 2a: 4-Chloro-6-ethyl-5-iodo-thieno[2,3-i/]pyrimidine

Step A: 6-Ethyl-3Y{-thieno[2,3-à]pyrimidin-4-one

The mixture of 701 g 2-amîno-5-ethyl-thiophene-3-carboxylic acid ethyl ester (3.52 mol) and 2200 mL formamide was heated to 200 °C and the lower boiling point solvents were distilled off. After 2 hours further 250 mL formamide was added and the mixture was stirred at the same température for another hour then at r.t. for 16 hours. The resulting mixture was poured into 7.5 L water and the precipitate was filtered off, washed with 1.5 L toluene and 3 L water then air dried to give the product as a brown crystalline solid. ’H NMR (500 MHz, DMSO-d₆) Ô: 12.40 (br s, IH), 8.05 (s, IH), 7.11 (t, IH), 2.85 (qd, 2H), l.27(t, 3H)

Step B: 6-Ethyl-5-iodo-3\A-thieno[2,3-à]pyrimidin-4-one

The mixture of 301 g 6-ethyI-3//-thieno[2,3-i/]pyrimidin-4-one (1.67 mol), 847 g iodine (3.34 mol), 1040 g silver sulfate (3.34 mol) and 1.7 L éthanol was stirred at r.t. for 3 days.

ORIGINAL

-39The resulting precipitate was filtered off and washed with éthanol (3 x 400 mL). The product was eluted from the filter cake with the following procedure: the filter cake was stirred with 800 mL DMF at 50 °C for l hour then the suspension was filtered. Tins sequence was repeated 6 times. The combined organic layer was evaporated to dryness to give the product as a tan crystalline solid.

Step C: Préparation 2a

The mixture of stirred 880 mL phosphorous oxychloride and 102 mL MN-dimethylaniline was heated to 95 °C and 220 g 6-ethyl-5-iodo-3//-thieno[2,3-tf]pyrimidin-4-one (0.719 mol) was added quickly at the same température and then stirred for 15 minutes. The reaction mixture was cooled to 80 °C and poured on a stirred mixture of water (l L), crushed ice (2 kg) and DCM (700 mL). The resulting mixture was stirred for further 30 minutes while the température was kept below 20 °C. The phases were separated, the inorganic layer was extracted with DCM (100 mL) and the organic layer was washed with water (100 mL). The combined organic layer was dried over MgSCL, filtered and the filtrate was concentrated under reduced pressure to give the product as a tan crystalline solid. H NMR (400 MHz, DMSO-d₆) δ: 8.79 (s, IH), 3.02 (q, 2H), 1.39 (t, 3H)

Préparation 2b: 5-Bromo-4-chloro-6-(4-fluorophenyl)thieno[2,3-i/]pyrimidine

75.08 g Préparation la (200 mmol), 53.63 g 2-(4-fluorophenyI)-4,4,5,5-tetramethyl-l ,3,2dioxaborolane (240 mmol), 130 g césium carbonate (400 mmol), 2.245 g Pd(OAc)2 (10 mmol) and 8.50 g 'BuX-Phos (20 mmol) were placed in a 2 L fiask. 600 mL THF and 200 mL water were added, and then stirred overnight at 70 °C under argon atmosphère. THF was evaporated, and then the product was collected by filtration. The crude product was sonicated in 250 mL acetonitrile and filtered again. Then Préparation 2h was crystalized from EtOH / THF (2:1). 'il NMR (400 MHz, DMSO-d₍,) δ: 9.02 (s, 1H), 7.80-

7.77 (m, 2H), 7.47-7.43 (m, 2H)

Préparation 2c: 4-Chloro-5-iodo-6-(prop-l-ynyl)-thieno|2,3-r/]pyriniidine

42.24 g Préparation 1b (100 mmol), 3.509 g Pd(PPh3)₂Cl (5 mmol) and 1.904 g Cul (10 mmol) were dissolved in 400 mL DIP A, then propyne was bubbled through the reaction mixture, which was stirred at r.t. until no further conversion was observed. The

ORIGINAL

-40volatiles were evaporated under reduced pressure and the crude product was purified via flash chromatography using heptane / EtOAc as eluents. ’H NMR (400 MHz, DMSO-dé) δ:

8.92 (s, l H), 2.25 (s, 3H)

Préparation 2d: 4-Chloro-5-iodo-6-isopropyI-thieno|2,3-i/]pyrimidine

Step A: 6-Isopropyl-5-iodo-3Y{-thieno[2,3-<3]pyrimidin-4-one

The mixture of 2.858 g (14.7 mmol) 6-isopropyl-3/7-thieno[2,3-i/]pyriniidin-4-one, 7.468 g (29.4 mmol) iodine, 9.175 g (29.4 mmol) silver sulfate, and 55 mL éthanol was stirred at r.t. for 3 days. The mixture was diluted with Et₂O, the resulting precipitate was filtered off and used without further purification.

‘H NMR (400 MHz, DMSO-d_f,) δ: 12.49 (br s, IH), 8.11 (s, IH), 3.35 (m, IH, overlapped by H₂O signal), 1.28 (d, 6H)

MS (M-H): 319.0

Step B: Préparation 2d

The mixture of 15 mL (161 mmol) phosphorous oxychloride and 1.9 mL (14.7 mmol) A/N-dimethylaniline was heated to 95°C and 25.9 g (14.7 mmol) 6-isopropyl-5-iodo-3/7thieno[2,3-iZ]pyrimîdin-4-one (0,719 mol) was added quickly and then stirred for further 15 minutes at this température. The reaction mixture was cooled to 80 °C and poured into a stirred mixture of icy water (300 g) and EtOAc (200 mL). The resulting mixture was stirred for further 30 minutes while the température was kept below 20 °C. The phases were separated, the inorganic layer was extracted with EtOAc (100 mL) and the organic layer was washed with water and NaHCO₃ solution. The combined organic layer was dried over MgSO4, filtered and the filtrate was concentrated under reduced pressure and purified via flash chromatography using heptane and EtOAC as eluents to give the title product.

’H NMR (400 MHz, CDC1₃) Ô: 8.78 (s, 1 H), 3.63 (septet, IH), 1.41 (d, 6H)

MS (M+H): 339.0

Préparation 3a: (2Æ)-2-[(6-ethyl-5-iodo-thieno[2,3-rf]pyriniidin-4-yI)amino]-3-phenylpropanoic acid

ORIGINAL

-4I Using General Procedure la and Préparation 2a as the appropriate 4-chloro-thieno[2,3-i7] pyrimidine dérivative and D-phenylalanine as the appropriate amino acid dérivative, Préparation 3a was obtained.

'H NMR (500 MHz, DMSO-d₆) Ô: 8.44 (s, IH), 7.45 (d, IH), 7.30-7.20 (m, 5H), 5.07 (m, l H), 3.35 (dd, lH),3.l6(dd, lH), 2.82 (q, 2H), 1.22 (t, 3H)

HRMS calculated for C_l7H_l6ÎN₃O₂S: 453.0008; found: 454.0064 (M+H)

Préparation 4a: 4-chloro-5-|3-chloro-2-methyl-4-[2-(4-niethylpiperazin-l-yl)ethoxy] phenyl]-6-iodo-thieno[2,3-r/]pyrimidine

Step A: 4-chloro-5-[3-chloro-2-methyl-4-[2-(4-methylpiperazin-î-yl)ethoxy]phenyl]thieno [2,3-djpyri mi d'n te

Using General Procedure lia and Préparation le as the appropriate 5-iodo-thieno[2,3-i/] pyrimidine dérivative and Préparation B4 as the appropriate boronic acid dérivative, and purifying the product via flash chromatography using DCM and MeOH as eluents gave 4-chloro-5-[3-chloro-2-methyl-4-[2-(4-methylpiperazin-l-yl)ethoxy]phenyl]thieno[2,3-i7] pyrimidine. 'H NMR (400 MHz, DMSO-d₆) 8: 8.98 (s, IH), 7.97 (s. IH), 7.22 (d, IH), 7.09 (s, IH), 4.25-4.16 (m, 2H), 2.76 (t, 2H), 2.54 (br s, 4H), 2.32 (br s, 4H), 2.14 (s, 3H), 2.06 (s, 3H)

Step B: Préparation 4a

10.935 g 4-chloro-5-[3-chloro-2-methyl-4-[2-(4-methylpiperazin-l-yl)ethoxy]phenyI] thieno[2,3-i/]pyrimidine (25 mmol) was dissolved in 250 mL dry THF and cooled to -78 °C. 25 mL LDA solution (50 mmol, 2M in THF, heptane, ethyl benzene) was added dropwise under Argon atmosphère and the mixture was stirred for 15 minutes. Then 12,69 g (50 mmol) iodine was added at -78°C and the mixture was allowed to warm up to r.t. Then the mixture was diluted with EtOAc and was washed with NH4CI solution then with Na₂S₂O₃ solution, dried over Na₂SO4, fîltered and concentrated under reduced pressure. The crude product was purified via flash chromatography using DCM and MeOH as eluents to obtain Préparation 4a. 'H NMR (500 MHz, DMSO-d₆) δ: 8.93 (s, IH), 7.15 (d, IH), 7.13 (d, IH), 4.22 (t, 2H), 2.77 (t, 2H), 2.56 (br s, 4H), 2.34 (br s, 4H), 2.16 (s, 3H), 2.00 (s, 3H)

ORIGINAL

-42Préparation 4b: 4-chIoro-5-[3-chIoro-2-methyl-4-[2-(4-methylpiperazin-l-yl)ethoxy] phenylJ-6-(2-furyl)thieno|2,3-rf] pyrimidine

Using General Procedure Ha and Préparation 4a as the appropriate 6-iodo-thieno[2,3-c/] pyrimidine dérivative and 2-(2-furyl)-4,4,5,5-tetramethyl-l,3,2-dioxaborolane as the appropriate boronic acid dérivative, Préparation 4b was obtained. MS: (M+H) = 503.0

Préparation 4c: 4-chloro-5-[3-chloro-2-methyl-4-[2-(4-methylpiperazin-I-yl)ethoxy) phenyl|-6-(5-fluoro-2-furyl)thieno|2,3-rf| pyrimidine

Using General Procedure Ilia and Préparation 4a as the appropriate 6-iodo-thieno[2,3-i7] pyrimidine dérivative and 2-(5-fluoro-2-furyl)-4,4,5,5-tetramethyl-l,3,2-dioxaborolane as

I0 the appropriate boronic acid dérivative, Préparation 4c was obtained.

'H NMR (500 MHz, DMSO-d₆) δ: 8.93 (s, 1H), 7.24 (d. IH), 7.18 (d, IH), 5.92 (dd, III),

5.68 (t, IH), 4.23 (t. 2H), 2.79 (t, 2H), 2.58 (br s, 4H), 2.38 (br s, 4H), 2.19 (s, 3H), 2.05 (s, 3H)

HRMS calculated for C24H23N4O2FSCI2: 520.0903; found: 521.0972 (M+H)

Préparation 4d: 2-chloro-4-(4-chloro-6-iodo-thieno[2,3-i/]pyrimidin-5-yl)-3-methyiphenol

Step A : [2-chloro-4-(4-chlorothieno[2,3-d]pyrimidin-5-yl)-3-methyl-phenoxy]triisopropyl-silane

Using General Procedure lia and Préparation le as the appropriate 5-iodo-thieno[2.3-c/] pyrimidine dérivative and Préparation B3 as the appropriate boronic acid dérivative, [2-chloro-4-(4-chlorothieno[2,3-tZ]pyrimidin-5-yl)-3-methyl-phenoxy]-triisopropyl-silane was obtained. 'H NMR (400 MHz. DMSO-d₆) δ: 8.95 (s, IH), 7.98 (s, IH), 7.13 (d, IH).

6.91 (d, 1 H), 2.05 (s, 3H), 1.40-1.29 (m, 3H), 1.10 (dd, 18H)

Step B: [2-chloro-4-(4-chîoro-6~ïodothieno[2,3-d]pyriniidin-5-yl)-3-methyl-phenoxy]~ triisopropyl-silane

33.7 g [2-chloro-4-(4-chlorothieno[2,3-i7]pyrimidin-5-yl)-3-niethyl-phenoxy]-triisopropylsilane (72 mmol) was dissolved in 300 mL dry THF and cooled to -78 °C. 43.2 mL LDA

ORIGINAL

-43 solution (86.4 mmol, 2M in THF, heptane, ethyl benzene) was added dropwise under Argon and the mixture was stirred for 15 minutes. Then 23.8 g iodine (93.7 mmol) was added at -78 °C and the mixture was allowed to warm up to r.t. Then the mixture was diluted with EtOAc and was washed with NH4CI solution then with Na2S2Ûj solution. The 5 organic layer was dried over Na2SÛ4, filtered and concentrated under reduced pressure.

’H NMR (400 MHz, DMSO-d₆) δ: 8.91 (s, IH), 7.05 (d, IH), 6.97 (d. IH), 1.99 (s, 3H), 1.39-1.30 (m, 3H), I.IO (dd, 18H)

Step C: Préparation 4d

10.0 g [2-chloro-4-(4-chloro-6-iodothieno[2,3-r/]pyriniidin-5-yl)-3-methyl-phenoxy]10 triisopropyl-silane (16.85 mmol) was dissolved in 100 mL dry THF and 18.5 mL TBAF solution (18.5 mmol, IM in THF) was added and the mixture was stirred at r.t. for 10 minutes. Then the mixture was diluted with EtOAc and washed with l M HCl solution and brine. The organic layer was dried over Na2SÛ4, filtered and concentrated under reduced pressure. The crude product was purified via flash chromatography using heptane 15 and EtOAc as eluents to obtain Préparation 4d. HRMS calculated for C13H7CI2IN2OS:

435.8701, found: 436.8780 (M+H)

Préparation 4e: 2-[2-chloro-4-(4-chloro-6-iodo-thienof2,3-rf]pyrimidin-5-yl)-3-methylphenoxy|-/V^V-dimethyl-ethanamine

Using General Procedure VI and Préparation 4d as the appropriate phénol dérivative and 20 AÇV-dimethylethanolamine as the appropriate alcohol, Préparation 4e was obtained. MS (M+H): 508.0

Préparation 4f: 2-chloro-4-|4-chloro-6-(3-thicnyl)thieno|2,3-i/]pyrimidin-5-yl]-3methyl-phenol

Using General Procedure 111a and Préparation 4d as the appropriate 6-iodo-thieno[2,3-c/] 25 pyrimidine dérivative and thiophene-3-boronic acid pinacol ester as the appropriate boronic acid dérivative. Préparation 4f was obtained. MS (M+H): 393.0

Préparation 4g: 4-chloro-5-|3-chloro-2-methyl-4-[2-(4-methylpiperazin-l-yl)ethoxy] phenyl|-6-(3-thienyl)thieno[2,3-rf] pyrimidine

ORIGINAL

-44Using General Procedure VI and Préparation 4f as the the appropriate 6-iodo-thieno [2,3-i7]pyrimidine dérivative and 2-(4-methylpiperazin-l-y!)ethanol as the appropriate alcohol, Préparation 4g was obtained.

’H NMR (500 MHz, DMSO-d₆) δ: 8.94 (s, IH), 7.60 (dd, IH), 7.56 (dd, IH), 7.19 (d, IH),

7.12 (d, IH), 6.79 (dd, IH), 4.21 (t, IH), 2.77 (t, IH), 2.56 (br, 4H), 2.33 (br, 4H), 2.15 (s, 3H), 2.04 (s, 3H)

HRMS calculated for C24H24CI2N4OS2: 518.0769; found: 519.0852 (M+H)

Préparation 4h: 4-[2-[2-chloro-4-[4-chIoro-6-(3-thienyl)thieno[2,3-rf]pyrimidin-5-yl|3-methyl-phenoxy]ethyl]morphoIine

Using General Procedure VI and Préparation 4f as the appropriate phénol dérivative and

2-morpholinoethanol as the appropriate alcohol, Préparation 4h was obtained.

Préparation 4i: 4-chloro-5-(l-naphthyl)thieno[2,3-r/|pyrimidine

Step A : ethyl 2-amino-4-(I-naphthyl)thiophene-3-carboxylate

50.00 g l-(l-naphthyl)ethanone (293.8 mmol), 43.66 g ethyl cyanoacetate (386.0 mmol), 18.84 g sulfur (587.5 mmol), 8.4 mL AcOH and 38.39 g morpholine were dissolved in 300 mL EtOH and stirred at 60 °C until no further conversion was observed. The volatiles were removed in vacuo, and the residue was purified via flash cromatorgaphy using heptane and EtOAc as eluents to obtain ethyl 2-amino-4-(l-naphthyl)thiophene-3carboxylate. HRMS calculated for C17H15NO2S: 297.0823; found: 298.0891 (M+H)

Step B: 5-(l-naphthyl)-3\A-thieno[2,3-d]pyrimidin-4-one

9.40 g ethyl 2-amino-4~(l-naphthyl)thiophene-3-carboxylate (31.6 mmol) was dissolved in 45 mL formamide and stirred at 200 °C until no further conversion was observed. The mixture was cooled to r.t. and poured into water. The precipitated solid was filtered, washed with water, then dried to obtain 5-(l-naphthyl)-3/7-thieno[2,3-i/| pyrimidin-4-one. HRMS calculated for C|(,IToNiOS: 278.0514; found: 279.0582 (M+H)

Step C: Préparation 4i

ORIGINAL

8.50 g 5-(I-naphthyl)-3//-thieno[2,3-i/]pyrimidin-4-one (30.5 mmol), 4.07 g AUV-dimethylaniline (33.6 mmol) and 22.8 mL phosphores oxychloride (244 mmol) were stirred at 100 °C for l hour. The mixture was cooled to r.t. and poured into stirred icy water. The precipitated solid was filtered and recrystallized from acetonitrile to obtain Préparation 4i. HRMS calculated for C|₆H₉N₂SCI: 296.0I75; found: 297.0255 (M+H)

Préparation 4j: 4-chloro-5-(3-chloro-2-methyl-phenyl)-6-ethyl-thieno|2,3-iZ] pyrimidine

Using General Procedure Ilb and Préparation 2a as the appropriate 5-iodo-thieno[2,3-<7] pyrimidine dérivative and (3-chloro-2-methyl-phenyl)boronic acid as the appropriate boronic acid dérivative, Préparation 4j was obtained.

'H NMR (400 MHz. DMSO-d₆) δ: 8.89 (s, IH), 7.55 (dd, IH), 7.33 (t, IH), 7.23 (dd, IH),

2.65 (m, 2H), 2.03 (s, 3H), 1.17 (t. 3H)

HRMS calculated for C15H12CI2N2S: 322.0098; found: 323.0164 (M+H)

Préparation 4k: 4-cliIoro-6-ethyl-5-(l-naphthyl)thieno|2,3-rfJpyrimidine

Using General Procedure Ilb and Préparation 2a as the appropriate 5-iodo-thieno[2,3-i/] pyrimidine dérivative and 1-naphthaleneboronic acid neopentyl glycol ester as the appropriate boronic acid dérivative, Préparation 4k was obtained.

‘H NMR (400 MHz, DMSO-d₆) δ: 8.91 (s, 1 H), 8.07 (dd, IH), 8.03 (dm. 1 H), 7.63 (dd, IH), 7.55 (tm, 1 H), 7.51 (dd, IH), 7.44 (tm, IH), 7.33 (dm, IH), 2.61 (q, 2H), 1.13 (t, 3H) HRMS calculated for C18H13CIN2S: 324.0488; found: 325.0562 (M+H)

Préparation 41: 4-chIoro-6-metliyl-5-(l-naphthyl)thieno[2,3-(/]pyrimidine

Using General Procedure IVa and methyl-iodide as the appropriate electrophile, Préparation 41 was obtained.

H NMR (400 MHz, DMSO-d₆) δ: 8.90 (s, IH), 8.04 (dd, 2H), 7.63 (dd, IH), 7.54 (td, 11-1),

7.49 (dd, IH), 7.43 (td, 1 H), 7.32 (d, IH), 2.28 (s, 3H)

MS (M+H): 311.0

Préparation 4m: [4-chloro-5-(l-naphthyl)thieno|2,3-rfjpyrimidin-6-yI|methanol

ORIGINAL

-46Sien A: 4-chloro-5-(l-naphthyl)thieno[2,3-à]pyrimidine-6-carbaldehyde

Using General Procedure IVa and DMF as the appropriate electrophile, 4-chloro-5-(lnaphthyi)thieno[2,3-<7]pyrimidine-6-carbaldehyde was obtained. 'H NMR (400 MHz. CDCl₃) δ: 9.65 (s, IH), 9.00 (s, IH), 8.07 (d, IH), 7.99 (d, IH), 7.68-7.52 (m, 3H), 7.47 (L IH), 7.33 (d, IH)

Sien B: Préparation 4m 4-chloro-5-(l-naphthyl)thieno[2,3-i/]pyrimidïne-6-carbaldehyde was dissolved in THF:MeOH l:l (4 mL/mmol) and 3 eq. NaBFIj was added at 0 °C. The mixture was stirred for 10 minutes, then quenched with IM citric acid. The mixture was extracted with DCM, washed with NaHCO₃ solution and brine, dried over Na₂SO₄, filtered and the filtrate was concentrated in vacuo. The crude product was purified via flash chromatography using heptane and EtOAc as eluents to obtain Préparation 4m.

'H NMR (400 MHz. DMSO-d₆) δ: 8.92 (s, IH), 8.06 (d. IH), 8.03 (d, IH), 7.62 (m, IH), 7.58-7.49 (m. 2H), 7.44 (m, l H), 7.35 (d. I H), 5.99 (t, l H), 4.54 (dd, l H), 4.33 (dd, l H) MS (M+H): 327.0

Préparation 4nl and Préparation 4n2: 1 - |4-chloro-5-( 1 -naphthyl)thieno|2,3-r/j pyrimidin-6-yl]ethanol

Using General Procedure IVa and acetaldehyde as the appropriate electrophilic reagent the crude product was obtained as a mixture of diastereoisomers that were separated by sequential flash chromatography using DCM-acetone and heptane-MTBE as eluents. The order of elution of the diastereomeric-pairs was the same in both eluent Systems, Préparation 4nl was obtained as the earlier eluting dîastereoisomer (racemate).

'H NMR (400 MHz, CDCI₃) δ: 8.85 (s, IH), 7.99 (d, IH), 7.95 (d, IH), 7.60-7.49 (m, 2H), 7.46-7.34 (m, 3H), 4.84 (m, 1 H), 2.06 (d, I H) 1.53 (d, 3H)

MS (M+H): 341.0

Préparation 4n2 was obtained as the later eluting dîastereoisomer (racemate).

'1-1 NMR (400 MHz, CDCI3) δ: 8.85 (s, IH), 7.99 (d, IH), 7.94 (d, IH), 7.60-7.49 (m, 2H),

7.46 (dd, 1 H) 7.43-7.37 (m, IH), 7.27 (overlap, IH), 4.98 (m, IH), 2.14 (d, IH) 1.35 (d, 3H)

MS (M+H): 341.0

ORIGINAL

-47Préparation 4o: l-[4-chloro-5-(l-naphthyl)thieno|2,3-rf|pyrimidin-6-yl]ethanone

157 mg Dess-Martin reagent (0.37 mmol) was dissolved in 2 mL DCM, then a mixture of Préparation 4nl and Préparation 4n2 (120 mg, 0.35 mmol dissolved in 10 mL DCM) was added and the mixture was stirred until no further conversion was observed. Then the mixture was diluted with DCM, washed with NaOH and NaHCO₃ solutions and brine, dried over Na₂SÛ4, filtered and concentrated under reduced pressure. The crude product was purified via flash chromatography using heptane and EtOAc as eluents to obtain Préparation 4o.

’H NMR (500 MHz, DMSO-d₆) δ: 9.09 (s, IH), 8.16 (dd. IH), 8.08 (d. IH), 7.72-7.65 (m, 2H), 7.62-7.57 (m, IH), 7.52-7.43 (m, 2H), l.7l (s, 3H)

MS (M+H): 339.0

Préparation 4p: 2-[4-chloro-5-(I-naphthyl)thieno[2,3-rf]pyrimidin-6-yl|propan-2-ol

Using General Procedure IVa and acetone as the appropriate electrophile, Préparation 4p was obtained.

'H NMR (400 MHz, CDCI₃) 8: 8.80 (s, IH), 7.98 (d. IH), 7.92 (d, IH), 7.59-7.46 (m, 2H), 7.46-7.34 (m, 2H), 7.30 (d, l H), 2.53 (br s, l H), l .54 (s, 3H), l .21 (s, 3H) MS (M+H): 355.0

Préparation 4q: 4-chloro-6-isopropyl-5-(l-naphthyl)thieno[2,3-rf|pyrÎmidine

Step A: 6-isopropyl-5-(l-naphthyl)-3W-ihieno[2,3-à]pyriinidin-4-one

250 mg Préparation 4p (0.705 mmol) and 1.75 mL Et₃SiH (10.9 mmol) were placed in a tlask and treated with 10 mL TFA at -10 °C. The mixture was then stirred at 50 °C until no further conversion was observed. The mixture was then diluted with DCM, neutralized with solid K₂CO₃ and NaHCO₃ solution. After séparation of the phases the organic phase was washed with brine, dried over Na₂SC>4, filtered and concentrated in vacuo to obtain 6-isopropyl-5-(l-naphthyl)-3/7-thieno[2,3-t/]pyrimidÎn-4-one as a crude intermediate. MS (M+H): 321.0

Step B: Préparation 4q

ORIGINAL mL phosphorous oxychloride and 0.161 mL Λ',Λ-dimethylaniline (l,27 mmol) were placed in a flask under Argon and 1.22 g 6-isopropyl-5-(l-naphthyl)-3H-thieno[2,3-i/] pyrimidin-4-one was added to the mixture in portions during 5 minutes. The reaction mixture was stirred at 100 °C until no further conversion was observed. The mixture was cooled to r.t. and poured into stirred icy water. The obtained aqueous media was neutralized by the careful addition of solid NaHCO₃. After the évolution of gas hâve ceased, the product was extracted three times with DCM. The combined organic layer was dried over Na₂SO4, filtered and concentrated under reduced pressure. The crude product was purified via flash chromatography using heptane and EtOAc as eluents to obtain

Préparation 4q.

'H NMR (500 MHz. CDCl₃) δ: 8.80 (s, IH), 7.97 (d, IH), 7.94 (d, IH), 7.57 (dd, IH), 7.54-7.49 (m, IH), 7.42-7.37 (m, 2H), 7.34 (d, IH), 3.02 (septet, IH), l.3l (d, 3H), 1.20 (d, 3H)

MS (M+H): 339.0

I5 Préparation 4r: 4-ehloro-6-(difluoromethyl)-5-(l-naphthyl)thicno[2,3-i/]pyrimidine 0.250 g 4-chloro-5-(l-naphthyl)thieno[2,3-</]pyrimidine-6-carbaldehyde (Step A intermediate in the synthesis of Préparation 4m, 0.77 mmol) was dissolved in 7 mL DCM, then 270 μί DAST (1.16 mmol) was added. The mixture was stirred at r.t. until no further conversion was observed. The mixture was then diluted with DCM and washed with water, then with NaHCO₃ solution and brine. The organic phase was dried over Na₂SO₄, filtered and concentrated under reduced pressure. The crude product was purified via flash chromatography using heptane and EtOAc as eluents to obtain Préparation 4r.

’H NMR (500 MHz, CDCl₃) δ: 8.97 (s, IH), 8.04 (d, IH), 7.97 (d, IH), 7.62-7.54 (m, 2H), 7.49-7.43 (m, 2H), 7.28 (d, l H), 6.47 (t, l H)

MS (M+H): 347.0

Préparation 4s: 4-chloro-6-iodo-5-(l-naphthyI)thieno|2,3-i/|pyrimidine

Using General Procedure IVa and iodine as the appropriate electrophilic reagent. Préparation 4s was obtained.

’H NMR (400 MHz, DMSO-d₆) δ: 8.94 (s, IH), 8.I0 (dm, IH), 8.05 (dm, IH), 7.66 (dm,

1 H), 7.56 (tm, 1 H), 7.48 (dd, 1 H), 7.44 (tm, 1 H), 7.31 (dm, 1 H)

ORIGINAL

-49HRMS calculated for Ci6H₈N₂SClI: 421.9141; found: 422.9211 (M+H)

Préparation 4t: 4-chloro-5-(3-chloro-2-methyI-phenyl)-6-iodo-thieno|2,3-i7]pyrimidine

Step A: 4-chloro-5-(3-chloro-2-methyl-phenyl)lhieno[2,3-à]pyrimidine

Using General Procedure Ilb and Préparation le as the appropriate 5-iodo-thieno[2,3-i/] pyrimidine derivative and (3-chloro-2-methyl-phenyl)boronic acid as the appropriate boronic acid derivative 4-chloro-5-(3-chloro-2-methyl-phenyl)thieno[2,3-t/]pyrimidine was obtained. ‘H NMR (400 MHz, CDC1₃) 6: 8.89 (s, IH), 7.47 (dd, IH), 7.43 (s, IH), 7.20 (t, IH), 7.14 (dd, IH), 2.14 (s, 3H)

Step B: Préparation 4i

Using General Procedure IVa and 4-chloro-5-(3-chloro-2-methyl-phenyl)thieno[2,3-t7] pyrimidine instead of Préparation 4i and iodine as the appropriate electrophilic reagent. Préparation 4t was obtained. ‘H NMR (400 MHz. CDCI₃) δ: 8.82 (s, IH), 7.52 (dd, IH), 7.25 (t, IH), 7.05 (dd, IH), 2.09 (s, 3H)

Préparation 4u: 4-chloro-5-(3-chloro-2-methyI-phenyl)-6-isopropyl-thieno[2,3-i/| pyrimidine

Using General Procedure Hb and Préparation 2d as the appropriate 5-iodo-thieno[2,3-J] pyrimidine derivative and (3-chloro-2-methyl-phenyl)boronîc acid as the appropriate boronic acid derivative, Préparation 4u was obtained.

’H NMR (400 MHz, DMSO-d_fi) δ: 8.90 (s, IH), 7.56 (dd, IH), 7.34 (t, IH), 7.29-7.22 (m, IH), 2.94 (septet, IH), 2.04 (s, 3H), 1.26 (d, 3H), 1.22 (d, 3H)

HRMS calculated for C_[6H₁₄N₂SC1₂: 336.0255; found: 337.0335 (M+H)

Préparation 4v: 4-chloro-6-ethyl-5-(lH-indol-4-yl)tliieno|2,3-rf|pyrimidine

The mixture of 0.664 g Préparation 2a (2.0 mmol), 0.400 g 1 /7-indol-4-ylboronic acid (1.2 eq, 2.4 mmol), 44.9 mg Pd(OAc)₂ (10 mol%, 0.2 mmol), 152 mg PCy₃xHBF₄(20 mol%, 0.4 mmol), 1.96 g Cs₂CO₃ (3.0 eq, 6.0 mmol) in 7.3 mL dimethoxyethane and

7.3 mL water was heated in microwave reactor at 100 °C until no further conversion was observed. The crude reaction mixture was fîitered through a pad of Celite, washed with

ORIGINAL

- 502 χ ΙΟ mL ΜΤΒΕ and 2 * ΙΟ mL water. The two layers of the filtrate were separated and the organic layer was washed with brine, then dried over Na₂SO4, filtered and concentrated in vacuo. The residue was purified via preparative reversed phase chromatography using water (containing O.l % TFA) and acetonitrile as eluents to obtain Préparation 4v.

'H NMR (400 MHz. DMSO-d₆) δ: 11.22 (br s, IH), 8.87 (s, IH), 7.49 (dm, IH), 7.32 (m, IH), 7.l9(dd, IH), 6.95 (dm, lH), 5.96 (m, lH), 2.67 (m, 2H), l.l4(t, 3H)

HRMS calculated for C_I6H,₂ClN₃S: 313.0440; found 314.0508 (M+H)

Préparation 4w: 4-chloro-5-(l-naphthyl)-6-vinyI-tliieno[2,3-i/]pyrimidîne

Using General Procedure Ile and Préparation 4s as the appropriate 6-iodo-thieno[2,3-i/] pyrimidine dérivative and vinylboronic acid pinacol ester as the appropriate boronic acid dérivative, Préparation 4w was obtained.

‘H NMR (500 MHz, DMSO-d₆) Ô: 8.95 (s, IH), 8.09 (d, IH), 8.05 (d, IH), 7.65 (dd, IH),

7.56 (t. IH), 7.52 (dd, IH), 7.45 (t, IH), 7.35 (d, IH), 6.34 (dd, IH), 5.90 (d, IH), 5.45 (d, IH)

HRMS calculated for C₁₈HhC1N₂S: 322.0331; found 323.0415 (M+H)

Préparation 4x: 4-chloro-5-(l-naphthyI)-6-|(.£7Z)-prop-l-enyl]thieno|2,3-i/|pyriinidine

Step A: 5,5-dimethyl-2-[(ZÆ,)-prop-1 -enyl]-l,3,2-dioxaborincme

To a solution of 0.172 g (Z)-prop-l-en-l-yl boronic acid (2.0 mmol, 9:1 ZIE isomer mixture) and 0.208 g neopentyl glycol (2.0 mmol) in 6 mL 2-Me-THF 20 mg Amberlyst 15H ionic exchange resin was added and it was stirred at r.t. until no further conversion was observed. The conversion was followed by 'lI-NMR measurement in CDC1₃ solution. The mixture was filtered through a pad of celite, washed with 2*3 mL 2-Me-THF and the filtrate was concentrated in vacuo. The resulting crude materiai was sufficiently pure for the next step as a 87:13 mixture of ZIE isomers according to NMR measurement. 'H NMR (400 MHz, CDCI3) δ: 6.57-6.43 (m, IH), 5.39-5.27 (dd, IH), 3.67 (s, 4H), 1.95-1.83 (dd, 3H), 0.97 (s, 6H)

Step B: Préparation 4x

ORIGINAL

- 51 Using General Procedure Ile and Préparation 4s as the appropriate 6-iodo-thieno[2,3-</| pyrimidine dérivative and 5,5-dimethyl-2-[(Z/£)-prop-l-enyl]-l,3,2-dioxaborinane (ZIEmixture, Step A) as the appropriate boronic acid dérivative, Préparation 4x was obtained as a 63:37 mixture of ZIE isomers.

’H NMR (500 MHz, DMSO-d₆) δ: 8.95-8.90 (s, IH), 8.H-8.06 (m, IH), 8.06-8.01 (m, IH), 7.67-7.60 (m, IH), 7.58-7.52 (m, IH), 7.52-7.48 (m, IH), 7.46-7.40 (m, IH), 7.36-

7.29 (m, IH), 6.45-5.90 (m, IH), 6.10-6.04 (m, IH), 2.06-1.72 (dd, 3H)

HRMS calculated for C|₉H|₃C1N₂S: 336.0488; found 337.0541 (M+H)

Préparation 4v: 4-chloro-6-isopropenyl-5-(l-naphthyl)thicno|2,3-r/|pyrimidine

Using General Procedure Ile and Préparation 4s as the appropriate 6-iodo-thieno[2,3-c/] pyrimidine dérivative and 2-isopropenyl-4,4,5,5-tetramethyM,3,2-dioxaborolane as the appropriate boronic acid dérivative, Préparation 4y was obtained.

'H NMR (500 MHz, DMSO-d₆) δ: 8.83 (s, IH), 7.96 (d, IH), 7.92 (d, IH), 7.55-7.37 (m, 5H), 5.23 (m, IH), 5.12 (m, IH), 1.65 (dd, 3H)

HRMS calculated for C|₉H|₃C1N₂S: 336.0488; found 337.0551 (M+H)

Préparation 4z: 4-chloro-5-(l-naphthyI)-6-|(£)-prop-l-enyl]thieno|2,3-i/| pyrimidine

Step A: 5,5-dimethyl-2-[(E)-prop-1 -enylJ-1,3,2-dioxaborinane

To a solution of 0.172 g (Æ)-prop-l-en-l-y] boronic acid (2.0 mmol) and 0.208 g neopentyl glycol (2.0 mmol) in 6 mL 2-Me-THF 20 mg Amberlyst 15H⁺ ionic exchange resin was added and it was stirred at r.t. until no further conversion was observed. The conversion was followed by 'lI-NMR measurement in CDC1₃ solution. The mixture was filtered through a pad of celite, washed with 2 ^χ 3 mL 2-Me-THF and the filtrate was concentrated in vacuo. The resulting crude material was sufficiently pure for the next step. It contained only the £-stereoisomer. ‘H NMR (400 MHz, CDCI3) δ: 6.57 (m, IH), 5.39 (dd, IH), 3.63 (s, 4H), 1.83 (dd, 3H), 0.97 (s, 6H)

Step B: Préparation 4z

ORIGINAL

-52Using General Procedure IlcIIIb and Préparation 4s as the appropriate 6-iodo-thieno [2,3-t7]pyrimidine dérivative and 5,5-dimethyl-2-[(E)-prop-l-enyl]-l,3,2-dioxaborinane (Step A) as the appropriate boronic acid dérivative, Préparation 4z was obtained.

‘H NMR (500 MHz, DMSO-d₆) δ: 8.90 (s, IH), 8.09 (d, IH), 8.04 (d, IH), 7.64 (dd, IH), 5 7.58-7.53 (m, IH), 7.50 (dd, IH), 7.44 (m, IH), 7.34 (d, IH), 6.45 (m, IH), 6.10-6.03 (m,

IH), 1.72 (dd, 3H)

HRMS calculated for C19H13CIN2S: 336.0488; found 337.0550 (M+H)

Préparation 5a: (2Æ)-2-|[5-(3-chloro-2-methyl-phenyl)-6-iodo-thicno|2,3-i/|pyrimidin4-yl]amino]-3-phenyl-propanoic acid

Using General Procedure Ib and Préparation 4t as the appropriate 4-chloro-thieno[2,3-t7] pyrimidine dérivative and D-phenylalanine as the appropriate amino acid dérivative, Préparation 5a was synthesized. The crude product was purified via préparative reversed phase chromatography using O.l % TFA solution and acetonitrile as eluents and Préparation 5a was obtained as a l : l mixture of diastereoisomers.

'H NMR (500 MHz, DMSO-d₆) δ: 13.15 (br s, IH), 8.42-8.41 (s, IH), 7.62-7.54 (d, IH), 7.39-7.17 (t. IH), 7.21-7.01 (m, d, IH), 7.21 (m, 4H), 6.82-6.79 (d, IH), 5.15-5.11 (d, IH), 4.82-4.76 (q, IH), 3.23-3.14 (dd, IH), 2.73-2.67 (dd, IH), 2.02-1.80 (s, 3H)

HRMS calculated for CizHn^CbSClI: 548.9775; found 549.9842 and 549.9864 (M+H)

Préparation 5b: (2/f)-2-|[(55_a)-5-[3-chIoro-2-methyl-4-[2-(4-methyIpiperazin-l-yl) 20 ethoxy]phenyI|-6-iodo-thieno[2,3-if]pyrimidin-4-yl]ainino|-3-|2-[(2-niethylpyrazoI-3yl)methoxy]phcnyl]propanoic acid

Using General Procedure Ib and Préparation 4a as the appropriate 4-chloro-thieno[2,3-i/] pyrimidine dérivative and Préparation Al as the appropriate amino acid dérivative, followed by HILIC purification. Préparation 5b was obtained as the later eluting 25 diastereoisomer. MS: (M+H) = 802.0

Préparation 5c: (2Æ)-2-[[5-[3-chloro-2-methyI-4-[2-(4-metliylpiperazin-l-yl)ethoxy| phenyl]-6-iodo-thieno|2,3-rf]pyrimidin-4-yl]amino|-3-[2-|(2-ethylpyrazol-3yl)methoxy]phenyl]propanoic acid

ORIGINAL

- 53 Using General Procedure Ib and Préparation 4a as the appropriate 4-chloro-thieno[2,3-i/] pyrimidine dérivative and Préparation A7 as the appropriate amino acid dérivative, followed by HILIC purification, Préparation 5c was obtained as the later eluting diastereomer. MS: (M+H) = 816.0

Préparation 6a: (27f)-2-[[5-bromo-6-(4-fluorophenyl)thieno[2,3-rf]pyriniidin-4-yl] amino]-3-(2-hydroxyphenyl)propanoic acid

Using General Procedure Ib and Préparation 2b as the appropriate 4-chloro-thieno[2,3-</] pyrimidine dérivative, and (2/?)-2-amino-3-(2-hydroxyphenyl)propanoic acid as the appropriate amino acid dérivative, Préparation 6a was obtained, isolated by filtration.

’H NMR (400 MHz. DMSO-d₆) 5: 12.90 (br s, IH), 9.65 (br s, 1H), 8.41 (s, IH), 7.70 (m, 2H), 7.45-7.34 (m, 3H), 7.18 (dd, IH), 7.04 (td, IH), 6.80 (d, IH), 6.72 (t, IH), 4.96 (m, IH), 3.31 (dd, lH), 3.08 (dd, IH) MS (M+H): 488.0

Préparation 6b: (2Æ)-3-(2-hydroxyphenyl)-2-|(5-iodo-6-prop-l-ynyl-thieno[2,3-d]

I5 pyrimidin-4-yI)amino]propanoic acid

Using General Procedure 1b and Préparation 2c as the appropriate 4-chloro-thieno[2,3-t7] pyrimidine dérivative, and (27?)-2-amino-3-(2-hydroxyphenyl)propanoic acid as the appropriate amino acid dérivative, Préparation 6b was obtained. The product was isolated by filtration instead of chromatography. MS: (M+H) = 480.0

Préparation 6c: methyl (2Æ)-2-|(6-ethyl-5-iodo-thieno|2,3-i/|pyrimidin-4-yl)aniino]-3phenyl-propanoate

3.246 g Préparation 2a (10 mmol), 3.70 g [(l/î)-l-benzyl-2-methoxy-2-oxo-ethyl] ammonium chloride (17 mmol) and 13.03 g Cs₂CO₃ (40 mmol) were dissolved in 15 mL DMSO and stirred at r.t. under N₂ atmosphère until no further conversion was observed.

The mixture was then acidified with 2M HCl solution to pH=l and extracted with 2 x 300 mL EtOAc. The combined organic phases were washed with NaHCO₃ solution, dried over Na₂SO₄, filtered and concentrated under reduced pressure. The crude product was purified via flash chromatography using heptane and EtOAc as eluents to obtain Préparation 6c.

ORIGINAL

-54’H NMR (500 MHz, DMSO-d₆) δ: 8.39 (s, IH), 7.33 (d, IH), 7.30 (m, 2H), 7.25-7.22 (m, 3H), 5.ll (m, IH), 3.69 (s,3H), 3.33 (dd, IH), 3.l8(dd, IH), 2.82 (q, 2H), 1.23 (t,3H) HRMS calculated for C18H1JN3O2S: 467.0164; found 468.0242 (M+H)

Préparation 7a: ethyl (2Æ)-2-f[5-|3-chloro-2-methyl-4-[2-(4-methylpiperazin-l-yl) ethoxy]phenyl]-6-(4-fluorophenyl)thieno|2,3-i/]pyrimidin-4-yl]amino]-3-(2hydroxyphenyl)propanoate

Step A: (2R)-2-[[5-[3-chloro-2-methyl-4-[2-(4-melhylpiperazin-l-yl)ethoxy]phenyl]-6-(4fluorophenyl)thieno[2,3-d]pyrimidin-4-yl]amino]-3-(2-hydroxyphenyl)propanoic acid

Using General Procedure Ild and Préparation 6a as the appropriate 5-iodo-thieno[2,3-iZ] pyrimidine dérivative and Préparation B4 as the appropriate boronic acid dérivative, (2Λ)-

2-[[5-[3-chloro-2-methyi-4-[2-(4-methylpiperazin-l-yl)ethoxy]phenyl]-6-(4-fluorophenyI) thieno[2.3-i/]pyrimidin-4-yl]amino]-3-(2-hydroxyphenyl)propanoic acid was obtained. HRMS calculated for C35H35CIFN5O4S: 675.2082; found 676.2097 (M+H)

Step B: Préparation 7a

2.3 g (2/?)-2-[[5-[3-chloro-2-methyl-4-[2-(4-methylpiperazin-l-yl)ethoxy]phenyl]6-(4-fluorophenyl)thieno[2,3-i/]pyrimidin-4-yl]amino]-3-(2-hydroxyphenyl)propanoic acid (3.4 mmol) was dissolved in 20 mL 1.25 M HCl in EtOH and stirred at 40°C overnight. The mixture was then diluted with NaHCOj solution and extracted with DCM. The combined organic phases were dried over Na₂SÛ4, filtered and concentrated in vacito. The crude product was purified via flash chromatography using DCM and MeOH as eluents to obtain Préparation 7a. HRMS calculated for C37H39CIFN5O4S: 703.2395; found 704.2417 (M+H)

Préparation 7ad2: ethyl (2Æ)-2-[|(5S^,„)-5-[3-chloro-2-methyl-4-(2-(4-methylpiperazinl-yl)ethoxy]phenyl]-6-(4-fluorophenyl)tliieno[2,3-i7|pyrimidin-4-yl]amino]-3-(2hydroxyphenyl)propanoate

Step A : (2^)-2-(((53^-5-(3-017101-0-2-1))01 hyl-4-[2-(4-methylpiperazin-1 -yl)ethoxy]phenyl]6-(4-fluorophenyl)thieno[2,3-à]pyrimidin-4-yl]amino]-3-(2-hydroxyphenyl)propanoic acid

ORIGINAL

-55Using General Procedure Tld and Préparation 6a as the appropriate 5-iodo-thieno[2,3-<7] pyrimidine dérivative and Préparation B4 as the appropriate boronic acid dérivative, (2Æ)-

2-[[5-[3-chloro-2-methyl-4-[2-(4-methyIpiperazin-l-yl)ethoxy]phenyl]-6-(4-fluorophenyl) thieno[2,3-(/]pyrinudin-4-yl]arnino]-3-(2-hydroxyphenyl)propanoic acid was obtained as a mixture of diastereomers. The mixture was separated via flash chromatography using HILIC eluents. The earlier eluting diastereoisomer was collected as Préparation 7a 1. MS (M+H): 676.2

The later eluting diastereoisomer was collected as Préparation 7a2. MS (M+H): 676.2

Step B: Préparation 7ad2

44.51 g of Préparation 7a2 (6.67 mmol) was dissolved in 85 mL 1.25 M HCl in EtOH and stirred at 40 °C overnight. The mixture was then cautiously diluted with NaHCCfe solution and extracted with DCM. The combined organic phases were dried over Na₂SO4, filtered and concentrated in vacuo. The crude product was purified via flash chromatography using DCM and MeOH as eluents to obtain Préparation 7ad2.

‘H NMR (500 MHz, DMSO-d₆) δ: 9.49 (s, IH), 8.40 (s, IH), 7.34 (d, IH), 7.27-7.21 (m, 3H), 7.20-7.14 (m, 2H), 7.00 (td, IH), 6.71 (dd, IH), 6.60 (td, IH), 6.39 (dd, IH), 5.03 (d, IH), 4.92 (m, IH), 4.26 (t, 2H), 4.03 (m, 2H), 3.03 (dd, IH), 2.78 (t, 2H), 2.54 (br, 4H),

2.36 (dd, IH), 2.30 (br, 4H), 2.12 (s, 3H), 1.83 (s, 3H), 1.10 (t, 3H)

HRMS calculated for C37H39CIFN5O4S: 703.2395; found 704.2450 (M+H)

Préparation 7b: ethyl (2R)-2-[[(55'_fl)-5-[3-chloro-2-niethyl-4-[2-(4-niethylpipcrazin-lyl)ethoxy|phenyl|-6-prop-l-ynyl-thieno|2,3-rf]pyrimidin-4-yl]amino]-3-(2hydroxyphenyl)propanoate

Step A: (2Sk)-2-[[(5S^-5-[3-chloro-2-methyl-4-[2-(4-methylpiperazin-l-yl)ethoxy]phenyi]6-prop-1 -ynyl-thieno[2,3-à]pyrimidin-4-yl]amino]-3-(2-hydroxy phenyl)propanoic acid

Using General Procedure Ilb and Préparation 6b as the appropriate 5-iodo-thieno[2,3-t/J pyrimidine dérivative and Préparation B4 as the appropriate boronic acid dérivative, Ataphos as catalyst and THF:water 3:1 as solvent a mixture diastereoisomers was obtained. They were separated via flash chromatography using HILIC eluents. The diastereoisomer eluting later was collected as (2Æ)-2-[[(5S„)-5-[3-chloro-2-methyl-4-[2-(4-rnethylpiperazinORIGINAL

-56I -y l)ethoxy]phenyl]-6-prop-1 -ynyl-thieno[2,3-c/]pyriniidin-4-yl|amino]-3-(2-hydroxy phenyl)propanoic acid. MS: (M+H): 620.2

Step B: Préparation 7b

2.3 g (2J?)-2-[[(55'_a)-5-[3-chloro-2-methyl-4-[2-(4-methylpiperazin-l-yl)ethoxy]phenyl] -6-prop-1 -ynyl-thieno[2,3-i/]pyrimidin-4-yl]amino]-3-(2-hydroxyphenyl)propanoic acid (3.71 mmol) was dissolved in 20 mL 1.25M HCl in EtOH and stirred at 40 °C overnight. The mixture was then diluted with NaHCCfe solution and extracted with DCM. The combined organic phases were dried over Na2SÛ4, filtered and concentrated in vacuo, The crude product was purified via flash chromatography using EtOAc or DCM and MeOH as eluents to obtain Préparation 7b.

‘H NMR (500 MHz, DMSO-d₆) δ: 9.47 (s, IH), 8.41 (s, IH), 7.21 (s, IH), 7.21 (s, IH), 7.00 (td, IH), 6.70 (dd, IH), 6.60 (td, IH), 6.34 (d, IH), 5.11 (d, IH), 4.89 (m, IH), 4.27 (t, 2H), 4.03 (m, 2H), 3.06 (dd, IH), 2.79 (t, 2H), 2.55 (br, 4H), 2.40 (dd, IH), 2.30 (br, 4H),

2.12 (s, 3H), 2.00 (s, 3H), 1.97 (s, 3H), l.l l (t, 3H)

HRMS calculated for C34H₃8ClN₅O₄S: 647.2333; found 648.2385 (M+H)

Préparation 7c: ethyl (2/?)-2-||5-[3-chloro-4-(2-dimethyIaminoethyloxy)-2-methylphenyi]-6-prop-l-ynyl-thieno[2,3-i/]pyrimidin-4-yl]aminoJ-3-(2-hydroxyphenyl) propanoate

Step A: (2B.)-2-[[(5S>_a)-5-[3-chloro-4-(2-dimethylaminoethyloxy)-2-inethyl-phenyl]-6-propI-ynyl-thieno[2,3-d]pyrhnidin-4-yl]amino]-3-(2-hydroxyphenyl)propanoic acid

Using General Procedure Ild and Préparation 6b as the appropriate 5-iodo-Îhicno]2,3-c/| pyrimidîne dérivative and Préparation B5 as the appropriate boronic acid dérivative, a mixture diastereoisomers was obtained. They were separated via flash chromatography using HILIC eluents. The diastereoisomer eluting later was collected as (2/?)-2-[[(5S’_tl)-5[3-chloro-4-(2-dimethylaminoethy[oxy)-2-methyl-phenyl]-6-prop-l-ynyl-thieno[2,3-i/] pyrimidin-4-yl]amino]-3-(2-hydroxyphenyl)propanoic acid. MS (M+H): 565.2

Step B: Préparation 7c

ORIGINAL

2.3 g (2Æ)-2-[[(5S'_a)-5-[3-chloro-4-(2-dimethylaminoethyfoxy)-2-methyl-phenyl]-6-prop-lynyl-thieno[2,3-iZ]pyrimidin-4-yl]amino]-3-(2-hydroxyphenyl)propanoic acid (4.07 mmol) was dissolved in 20 mL l .25M HCl in EtOH and stirred at 40 °C ovemight. The mixture was then diluted with NaHCCfe solution and extracted with DCM. The combined organic phases were dried over Na^SO-j, filtered and concentrated in vacuo. The crude product was purified via flash chromatography using DCM and MeOEI as eluents to obtain Préparation 7c.

‘H NMR (500 MHz, DMSO-d₆) δ: 9.45 (s, IH), 8.41 (s, IH), 7.21 (s, IH), 7.21 (s, IH), 7.00 (td, IH), 6.70 (dd, IH), 6.60 (td, IH), 6.34 (d. IH), 5.12 (d, IH), 4.89 (m, IH), 4.26 (m, 2H), 4.03 (m, 2H), 3.06 (dd, IH), 2.74 (t. 2H), 2.39 (dd, IH), 2.27 (s, 6H), 2.01 (s, 3H), l.97(s, 3H), l.ll (t,3H)

HRMS calculated for Q^CIN^S: 592.1911; found 593.1954 (M+H)

Préparation 7d: ethyl (2Æ)-2-f[5-bromo-6-(4-fluorophenyl)thieno[2,3-r/]pyriniidin-4yl]amino]-3-(2-hydroxyphenyl)propanoate

2.5 g of Préparation 6a (5.1 mmol) was dissolved in 20 mL 1.25M HCl in EtOH and stirred at 40 °C ovemight. The resulting mixture was diluted with aq. NaHCOj solution and extracted with DCM. The combined organic phases were dried over Na^SO.^ filtered and concentrated in vacuo. The crude product was purified via flash chromatography using heptane and EtOAc as eluents to obtain Préparation 7d. ’H NMR (400 MHz, DMSO-d₆) δ: 9.67 (s, 1 H), 8.42 (s, IH), 7.70 (m, 2H), 7.43-7.37 (m, 3H), 7.14 (dd, IH), 7.05 (td, IH),

6.80 (dd, 111), 6.72 (td, IH), 5.01 (m, IH), 4.12 (q, 2H), 3.26 (dd, IH), 3.14 (dd, IH), 1.17 (t, 3H)

Préparation 7e: mcthyl (2/î)-2-| [6-ethyl-5-(4-hydroxy-2-methyl-phenyl)thieno|2,3-i/| pyrimidin-4-yl]amino|-3-phenyl-propanoate

934 mg Préparation 6c (2 mmol), 903 mg Préparation B6 (2.4 mmol), 231 mg Pd(PPli3)₄(0.2 mmol), 662 mg Ag2CÛ3 (2.4 mmol) and 81 pL methanol (2 mmol) were dissolved in 20 mL 2-Me-THF and stirred in MW reactor at 110 °C until no further conversion was observed. The mixture was filtered through Celite, diluted with 100 mL EtOAc then

2.5 mL TBAF (IM solution in THF) was added and the mixture was stirred at r.t. until no further conversion was observed. The mixture was then washed with NH₄C1 solution and

ORIGINAL

-58brine, dried over Na₂SC>4, filtered and concentrated and purified via flash chromatography using heptane and EtOAc as eluents to obtain Préparation 7e as a mixture of diastereoisomers.

'H NMR (400 MHz, CDCI₃) δ: 8.43-8.43 (s, IH), 7.26-6.80 (m, 7H), 6.76-6.64 (m. 2H), 5 5.18 (m, IH), 5.03 (m, IH), 3.66-3.65 (s, 3H), 3.16-3.13 (dd, IH), 2.73 (dd, IH), 2.57 (m,

2H), 2.07-1.80 (s, 3H), 1.18-1.17 (t, 3H)

MS (M+H): 448.2

Préparation 7f: methyl (2Zf)-2-[ |5-(3,5-dichloro-4-hydroxy-2-methyl-phenyl)-6-ethylthieno|2,3-rf]pynmidin-4-yl]amino]-3-phenyl-propanoate

I0 402 mg Préparation 7e (0.898 mmol) and 300 mg NCS (2.245 mmol) were dissolved in 5 mL THF and stirred at 60 °C until no further conversion was observed. The volatiles were removed in vacuo, the residue was purified via flash chromatography using heptane and EtOAc as eluents to obtain the title product as a mixture of diastereoisomers.

‘H NMR (500 MHz. DMSO-d₆) δ: 10.46-10.44 (s, IH), 8.40-8.38 (s, IH), 7.29-7.24 (s,

IH), 7.20 (m, 3H), 6.80-6.78 (d, 2H), 5.09-5.01 (d, IH), 4.95 (m, IH), 3.59-3.58 (s, 3H), 3.15-3.13 (dd, IH), 2.78-2.61 (dd. 1H),2.53 (q, 2H), 2.02-1.84 (s,3H), 1.11 (t, 3H) HRMS calculated for C₂5H₂3C1₂N3O₃S: 515.0837; found 516.0908 (M+H)

Préparation 7g: methyl (2/f)-2-[|5-(3-chloro-4-hydroxy-2-methyI-phenyl)-6-ethylthÎeno[2,3-if]pyrimidÎn-4-yl]amino]-3-phenyl-propanoatc

Using General Procedure 11c and Préparation 6c as the appropriate 5-iodo-thieno[2,3-c7] pyrimidine dérivative and Préparation B2 as the appropriate boronic acid dérivative, Préparation 7g was obtained as a mixture of diastereoisomers. MS (M+H): 482.1

Préparation 7gdl: methyl (27î)-2-[|(55_n)-5-(3-chloro-4-liyilroxy-2-niethyl-phenyl)-6Ηΐιν1-ίΙΐΗ·ηο[2,3-//| py rimidin-4-yl|aniino|-3-phenyl-propanoate

The diastereoisomers of Préparation 7g were separated via flash chromatography using heptane and EtOAc as eluents. The diastereoisomer eluting later was collected as Préparation 7gdl.

ORIGINAL

-59’H NMR (500 MHz. DMSO-d₆) δ: 10.53 (s, IH), 8.36 (s, IH), 7.23 (m, 2H), 7.20 (m, IH), 7.04 (d, IH), 6.98 (d. IH), 6.80 (m, 2H), 5.11 (d, IH), 4.90 (m, IH), 3.57 (s, 3H), 3.10 (dd, IH), 2.63 (dd. IH), 2.51-2.46 (m, 2H), 1.86 (s, 3H), I.IO (t, 3H)

HRMS calculated for C₂₅H₂4ClN₃O₃S: 481.1227; found 482.1313 (M+H)

Préparation 7h: (2Æ)-2-[[6-ethyl-5-(17/-indol-4-yl)thieno[2,3-i/]pyriniidin-4-yl|amino|3-phenyl-propanoic acid

Using General Procedure Ib and Préparation 4v as the the appropriate 4-chlorothieno[2,3-i/]pyrimidine dérivative and (2/?)-2-amino-3-(2-hydroxyphenyl)propanoic acid as the appropriate amino acid dérivative, Préparation 7h was obtained as a mixture of diastereoisomers.

‘H NMR (500 MHz, DMSO-d_(ï) δ: I2.7l-l2.59 (br s, IH), 11.48-11.37 (s, IH), 8.35-8.30 (s, IH), 7.64-7.53 (d, IH), 7.45-7.39 (dd, IH), 7.30-7.08 (t, IH), 7.17-6.33 (m, 6H), 6.076.01 (s, IH), 5.27 (d, IH), 4.59/4.50 (m, IH), 2.98-2.83 (dd, IH), 2.56 (m, 2H), 2.35-2.15 (dd, IH), 1.11-1.09 (t, 3H)

HRMS calculated for C25H₂₂N4O₂S: 442.1463; found 443.1529 and 443.1538 (M+H)

Préparation 7i: methyl (2Æ)-2-[[5-(3-chloro-lH-indol-4-yl)-6-ethyl-thieno|2,3-i/| pyrimidin-4-yI]aminoJ-3-phenyl-propanoatc

Step A: methyl (2R)-2-[[6-elhyl-5~(l\\-indol-4-yl)thieno[2.3-dlpyrimidin-4-yl]amin()]-3phenyl-propanoate

8.87 g Préparation 7h (20 mmol) was dissolved in 60 mL MeOH and 5.88 mL cc. H₂SO4 (60 mmol) was added. The mixture was stirred at r.t. under N₂ atmosphère for 2 hours. The mixture was then poured into icy water, the precipitate was fîltered to obtain methyl (27?)2-[[6-ethyl-5-(l/7-indol-4-yl)thieno[2,3-i/]pyrimidin-4-yl]amino]-3-phenyl-propanoate as a mixture of diastereoisomers.

‘H NMR (400 MHz, DMSO-d₆) δ: 11.52-11.43 (s, IH), 8.39-8.34 (s, IH), 7.65-7.57 (d, IH), 7.47-7.42 (t, 1 H), 7.30-7.11 (dd, IH), 7.18-6.79 (m, 2H), 7.02 (m, IH), 6.93 (m, IH),

6.65 (m, IH), 6.34 (m, IH), 6.05 (dt, IH), 5.28 (m, IH), 4.71-4.62 (m, 1 H), 3.55-3.41 (s, 3H), 2.91-2.77 (dd, IH), 2.57 (m, 2H), 2.37-2.23 (dd, IH), 1.11-1.10 (t, 3H) MS (M+H): 457.2 and 457.2

ORIGINAL

-60Siep B: Préparation 7/

8.477 g methyl (2Æ)-2-[[6-ethyl-5-(I/7-indol-4-yl)thieno[2,3-i/]pyrimidin-4-yl]amino]-3phenyl-propanoate (18.5 mmol), 2.47 g NCS (18.5 mmol) and 30 mL abs. THF were stirred at r.t. until no further conversion was observed. Then the mixture was poured into icy water and was extracted with EtOAc. The combined organic phases were dried over Na₂SO₄, filtered and concentrated under reduced pressure. The crude product was purified via flash chromatography using heptane and EtOAc as eluents to obtain Préparation 7i as a mixture of diastereoisomers.

'H NMR (500 MHz, DMSO-d₆) δ: 11.73-11.65 (d. IH), 8.35-8.31 (s, IH), 7.63-7.56 (d. IH), 7.62-7.54 (d, IH), 7.44-7.15 (dd. IH), 7.20-7.03 (m, 3H), 7.04-6.84 (d, IH), 6.70-6.44 (dm, 2H), 5.09-4.98 (d, IH), 4.80-4.72 (m, IH), 3.51-3.38 (s, 3H), 2.93-2.81 (dd, IH), 2.52 (m, 2H), 2.46-2.29 (dd, IH), 1.10-1.09 (t, 3H)

HRMS calculated for C₂₆H₂₃C1N₄O₂S: 490.1230; found 491.1282 and 491.1316 (M+H)

Préparation 7j: 4-chIoro-5-(3-chloro-l/Z-indol-4-yl)-6-ethyl-thieno[2,3-rf]pyrimidine

The mixture of 1.099 g Préparation 4v (3.5 mmol) and 0.572 g NCS (4.2 mmol) in 20 mL CCI4 was stirred at r.t. until no further conversion was observed. The mixture was then poured onto crushed ice, and it was extracted with DCM. The combined organic layers were dried over Na₂SO₄, filtered and concentrated under reduced pressure. The crude product was purified via flash chromatography using heptane and EtOAc as eluents to obtain 4-chloro-5-(3-chloro-l /7-indol-4-yl)-6-ethyl-thieno[2.3-J]pyrimidine Préparation 7j. ’H NMR (400 MHz, CDCI3) δ: 8.79 (s, IH), 8.33 (br s, IH), 7.47 (dd, IH), 7.31 (t, IH), 7.18 (d, IH), 7.03 (dd, IH), 2.73 (m, 2H), 1.24 (t. 3H)

Préparation 8a: (£)-4-( Diniethylamino)-l.l-dirnethoxv-but-3-en-2-one

502.1 g l,l-dimethoxypropan-2-one (4.25 mol) and 506.4 g 1,1-dimethoxy-MY-dimethylmethanamine (4.25 mol) were mixed in a 2 L flask and stirred at 105 °C for 3 hours. The formed MeOH was removed continuously via distillation. When MeOH formation stopped (at 65 °C head température) the reaction mixture was vacuum distilled (decreasing the pressure slowly to 30 mbar) to remove side products and unreacted starting materials. The crude product was distilled at 0.1 mbar. Fractions were collected between 107-118 °C head

ORIGINAL

-6l · température (bath température 160-I65 °C) to give a yellow oil. ^!H NMR (500 MHz, DMSO-dô) δ: 7.59 (d, IH), 5.17 (d, IH), 4.42 (s, IH), 3.25 (s, 6H), 3.09 (s, 3H), 2.78 (s, 3H)

Préparation 8b: 4-(Dimethoxyniethyl)-2-methylsulfonyl-pyrimidine

Step A: 4-(Dhnethoxymelhyl)-2-methylsulfanyl-pyrimidine

198 g sodium methoxide (3.67 mmol) was dissolved in 3 L MeOH and cooled to 0 °C. 322 g thiocarbamide (4.23 mol) was added portionwise and the mixture was stirred for l hour. Then 488 g Préparation 8a (2.82 mol) was added dropwise at 0 °C, then it was heated to 70°C until no further conversion was observed. It was cooled to r.t., 237 mL methyl iodide (3.81 mol) was added dropwise, keeping the température below 28 °C, and the resulting mixture was stirred overnight at r.t. It was filtered, the filtrate was concentrated under reduced pressure, diluted with EtOAc, washed with water and brine. The combined aqueous layers were extracted with EtOAc. The combined organic layers were dried over Na₂SÛ4, filtered and concentrated under reduced pressure. The residue was dissolved in 500 mL Et₂O, filtered through a pad of siiica, using Et₂O as eluent. The filtrate was concentrated under reduced pressure to give a light brown oil. *H NMR (400 MHz. DMSO-d₆) δ: 8.69 (d, IH), 7.23 (d, IH), 5.22 (s, IH), 3.33 (s, 6H), 2.52 (s, 3H)

Step B: Préparation 8b

To a solution of 180 g 4-(dimethoxymethyl)-2-methylsulfanyl-pyrirnidine (940 mmol) in

1.5 L methanol and 1.5 L water 752 g Oxone® (potassium peroxymonosulfate, 1220 mmol) was added portionwise al -5 °C, then stirred at 0 °C overnight. The reaction mixture was concentrated under reduced pressure to half volume using a 30 °C bath and then the mixture was filtered, and the precipitate was washed with DCM. The filtrate was extracted with DCM. The combined organic layers were dried over MgSCh, filtered and concentrated under reduced pressure to give a light brown oil. ’l-l NMR (400 MHz, CDCl₃) Ô: 8.98 (d, IH), 7.97 (d, IH), 5.36 (s, IH), 3.47 (s, 6H), 3.39 (s, 3H)

Préparation 9a: methyl (2R)-2-amino-3-(2-hy<Jroxypiienyl)propanoate hydrocliloride

ORIGINAL

24.6 g (27î)-2-amino-3-(2-hydroxyphenyl)propanoic acid (136 mmol) was stirred at r.t. in 900 mL solution of 3M HCl in methanol for 40 hours. The reaction mixture was concentrated under reduced pressure keeping the bath température below 40 °C. The residue was triturated with diethyl ether to give the product as a cream colored shining powder. HRMS calculated for C11I-L5NO3 (free base): 209.1052; found 210.1128 (M+H)

Préparation 9b: methyl (2Æ)-2-(/erf-butoxycarbonylamino)-3-(2-hydroxyplienyl) propanoate

16.7 g Préparation 9a (73.0 mmol) was suspended in 180 mL DCM. 30.5 mL (219 mmol) TEA was added and the solution was cooled using a water-ice bath. A solution of 15.6 g di-/i?/7-butyl bicarbonate (73.0 mmol) in 75 mL DCM was added slowly (2.5 hours). The mixture was stirred overnight at r.t. Then 100 mL water was added and the organic phase was separated, washed with water, IM HCI solution and finally with water again. The organic phase was dried over Na₂SO₄, filtered and the filtrate was concentrated under reduced pressure to obtain the product as an oil.

Préparation Al: (27î)-2-amino-3-|2-|(2-methylpyrazol-3-yl)methoxy]phenyl] propanoic acid

Using General procedure IXa and (2-methyIpyrazol-3-yl)methanol as the appropriate alcohol dérivative, Préparation Al was obtained. MS (M+H): 276.2

Préparation A2: (27f)-2-amino-3-|2-[(2-ethoxypyrimidin-4-yl)methoxy]phenyl| propanoic acid

Using General procedure IXa and Préparation Cl as the appropriate alcohol dérivative, Préparation A2 was obtained. MS (M+H): 318.1

Préparation A3: (2/f)-2-amino-3-|2-|(2-butylpyrazol-3-yl)methoxy]phenyl]propanoic acid

Using General procedure IXa and Préparation C2 as the appropriate alcohol dérivative, Préparation A3 was obtained. MS (M+H): 318.2

ORIGINAL

- 63 Préparation A4: (2Æ)-2-amino-3-[2-| [2-(2-methoxy pheny l)py rimidin-4-y l] methoxy | phcnyljpropanoic acid

Using General procedure IXa and Préparation C3 as the appropriate alcohol dérivative, Préparation A4 was obtained. MS (M+H): 380.2

Préparation A5: (2/î)-2-amino-3-[2-(2-pyridylmethoxy)phenyl]propanoic acid

Using General procedure IXa and 2-pyridylmethanol as the appropriate alcohol dérivative, Préparation A5 was obtained. MS (M+H): 273.1

Préparation A6: (2Zf)-2-amino-3-[2-(2,2,2-trifluoiOetlioxy)phenyl]propanoic acid

Using General procedure IXb and 2,2,2-trifluoroethyl trifluoromethanesulfonate as the appropriate alkylating reagent. Préparation A6 was obtained. MS (M+H): 264.1

Préparation A7: (2Æ)-2-amino-3-[2-[(2-ethylpyrazoI-3-yl)methoxy]phenyl]propanoic acid

Using General procedure IXa and (2-ethylpyrazol-3-yl)methanol as the appropriate alcohol dérivative, Préparation A7 was obtained. HRMS calculated for CuHtgNjOa: 289.1426, found: 290.1512 (M+H)

Préparation A8: (2/f)-2-amino-3-|2-[ j2-(2,2,2-trifIuoroethoxy)pyrimidin-4-yl] methoxy|phcnyljpropanoic acid

Using General procedure IXa and Préparation C8 as the appropriate alcohol derivalive, Préparation A8 was obtained. MS (M+H): 372.1

Préparation A9: (2/f)-2-amino-3-j2-j2-(dimethylaniino)-2-oxo-ethoxy]phenyl] propanoic acid

Using General procedure IXb and 2-chloro-A,A-dimethylacetaniide as the appropriate alkylating reagent, Préparation A9 was obtained. MS (M+H): 267.1

Préparation A10: (2Jf)-2-amino-3-[2-(2-cyclopentylcthoxy)phenyljpropanoic acid

Using General procedure IXa and 2-cyclopentylethanol as the appropriate alcohol dérivative. Préparation A10 was obtained. MS (M+H): 278.2

ORIGINAL

-64Préparation Ail: (2Æ)-2-amino-3-(2-phenethyloxyphenyI)propanoic acid hydrochloride

Using General procedure IXa and 2-phenylethanol as the appropriate alcohol dérivative, Préparation Al 1 was obtained. MS (M+H): 286.1

Préparation A12: (2/t)-2-amino-3-[2-(3-phenylpropoxy)phenyl| propanoic acid

Using General procedure IXa and 3-phenylpropan-l-ol as the appropriate alcohol dérivative, Préparation Al2 was obtained. MS (M+H): 300.2

Préparation A13: (27i)-2-amino-3-[2-((3-chlorophenyl)methoxy|phcnyl|propanoic acid

Using General procedure IXa and (3-chlorophenyl)methanol as the appropriate alcohol dérivative. Préparation A13 was obtained. MS (M+H): 306.1

Préparation A14: (2Æ)-2-amino-3-|2-[2-(4-methylpiperazin-l-yl)ethoxy|phenyl| propanoic acid

Using General procedure IXa and 2-(4-methylpiperazin-l -yl)ethanol as the appropriate alcohol dérivative, Préparation A14 was obtained. MS (M+H): 308.2

Préparation A15: (2/f)-2-amino-3-|2-(2-dimcthyianünoethyloxy)phenyl]propanoic acid

Using General procedure IXa and 2-(dimethylamino)ethanol as the appropriate alcohol dérivative, Préparation A15 was obtained. MS (M+H): 253.2

Préparation A16: (2Æ)-2-amino-3-|2-[3-(dimethylamino)propoxy]phenyl|propanoic acid

Using General procedure IXa and 3-(dimethylamino)propan-l-ol as the appropriate alcohol dérivative. Préparation Al6 was obtained. MS (M+H): 267.2

Préparation Bl: 3-methyl-4-(3,3,4,4-tetraniethylborolan-l-y!)-lH-indole

ORIGINAL

-65 l .87 g 4-bromo-3-methyl-l7/-indole (8.9 mmol), 5.028 g bis(pinacolato)diboron (19.6 mmol), and 2.65 g potassium acetate (26.7 mmol) were dissolved in 35 mL dry DMF under Argon, then 652 mg [l,l'-bis(diphenylphosphino)ferrocene]dichloropalladium(n) (0.89 mmol) was added. The reaction mixture was heated to 85 °C and stirred until no 5 further conversion was observed. Then it was concentrated under reduced pressure and purified via flash chromatography using heptane and EtOAc as eluents to obtaîn Préparation Bl.

'H NMR (400 MHz, CDCl₃) δ: 7.92 (br s, IH), 7.56 (d, IH), 7.42 (dd, IH), 7.16 (t. IH), 7.01 (d, IH), 2.47 (d, 3H), 1.40 (s, 12H)

I0 HRMS calculated for C|₅H₂₀NO₂B: 257.1587; found 258.1665 (M+H)

Préparation B2: 2-Chloro-3-methyl-4-(4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2-yl) phénol

Step A : (4-Brotno-2-chloro-phenoxy)-trimeihyl-silane

20.8 g 4-bromo-2-chloro-phenol (100 mmol) was dissolved in 150 mL dry THF then 15 24.2 g HMDS (150 mmol) was added. The reaction mixture was stirred at 85 °C under argon atmosphère for 1.5 hours then concentrated under reduced pressure. The resulted crude product was used without further purification. ’H NMR (200 MHz, CDC1₃) δ: 7.49 (d, 1 H), 7.23 (dd, 1 H), 6.75 (d, 1 H), 0.26 (s, 9H)

Step B: 4-Bromo-2-chloro-3-melhyl-phenol

48 mL ⁿBuLi solution (120 mmol, 2.5 M in hexanes) was added dropwise to a solution of

12.1 g dry DIPA (120 mmol) in 250 mL dry THF at -78 °C under argon atmosphère. The mixture was stirred for 30 minutes at the same température then 28.0 g (4-bromo-2-chlorophenoxy)-trimethyl-silane (100 mmol) was added dropwise. After 2.5 hours 21.3 g Mel (150 mmol) was added dropwise then the cooling bath was removed and the mixture was stirred overnight. The reaction was quenched with 100 mL NH4OH solution and 200 mL NH4CI solution then extracted with EtOAc. The organic phase was dried over Na₂SÛ4, fîitered and the filtrate was concentrated under reduced pressure. The resulting dark mass was refluxed with pure hexane several limes (150-150 mL aliquots) and decanted leaving a black tar behind. The combined organic phases were concentrated under reduced pressure

ORIGINAL

- 66 affording 19.0 g crude product, which was used without further purification. 'H NMR (200 MHz, CDCb) δ: 7.32 (d, IH), 6.76 (d, IH), 5.62 (s, IH), 2.49 (s, 3H)

Step C: (4-Broino-2-chloro-3-methyl-phenoxy)~trimethyl-silane

20.8 g HMDS (129 mmol) was added to the solution of 19.0 g 4-bromo-2-chloro-3-methylphenol (86.0 mmol) in 150 mL dry THF. The mixture was stirred at 85 °C under argon balloon for 1.5 hours and then concentrated under reduced pressure. The obtained product was used without further purification. *H NMR (200 MHz, CDCb) δ: 7.30 (d. IH), 6.63 (d,

H), 2.50 (s, 3H), 0.28 (s, 9H)

Step D: Préparation B2

A solution of 25.2 g (4-bromo-2-chloro-3-methyl-phenoxy)-trimethyl-silane (86.0 mmol) in 250 mL dry THF was cooled to -78 °C under argon and then 38 mL ⁿBuLi solution (94.6 mmol, 2.5M in hexanes) was added dropwise. After 5 minutes 19.2 g 2-isopropoxy-

4,4,5,5-tetramethyl-l,3,2-dioxaborolane (103 mmol) was added dropwise. The cooling bath was removed and the mixture was slowly allowed to warm up to r.t. Then the mixture was added to 200 mL NH4CI solution and extracted with EtOAc. The combined organic layers were concentrated under reduced pressure and passed through a pad of silica gel using hexane and EtOAc as eluents. The crude product was recrystallized from a mixture of EtOAc and hexane to obtain Préparation B2. 'H NMR (500 MHz, DMSO-d(,) Ô: 10.40 (s, IH), 7.42 (d, IH), 6.80 (d, lH), 2.49 (s,3H), l.27(s, 12H)

Préparation B3: |2-Chloro-3-methyl-4-(4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2-yl) phenoxy|-triisopropyl-silane

Step A: (4-Bromo-2-chloro-phenoxy)~triisopropyl-silane

200 g 4-bromo-2-chloro-phenol (0.97 mol) and 126 mL TIPSCI (I.l8 mol) were dissolved in 1.6 L DCM. 167 g imidazole (2.45 mol) was added and the mixture was stirred at r.t. for hours. Then the volatiles were evaporated under reduced pressure and the residue was dissolved in 1.5 L EtOAc. The mixture was washed with brine, dried over Na₂SÛ4, filtered and the filtrate was concentrated under reduced pressure. The triisopropylsilyl hydroxide impurity was removed by distillation (120 °C at 0.01 mmHg). The residue was filtered

ORIGINAL

-67through a short pad of silica with hexane and concentrated under reduced pressure. The product (colourless oil) was used in the next step without further purification.

’H NMR (400 MHz, CDCI3) 6: 7.49 (d, IH), 7.21 (dd, 1H),6.78 (d, IH), l.3l (septet, 3H),

I.l4 (d, 18H)

MS (El, 70 eV) m/z (% relative intensity, [ion]): 63 (30), 79 (24), 93 (41), 170 (17), 235 ( 19), 251 ( 16), 265 (24), 293 (23), 319 (77), 321 ( 100), 323 (28), 362 ( l, [M⁺]).

Step B: (4-Bromo -2-chloro-3 -methyl-phenoxy)-tri isopropyl-sil a ne

76.0 mL dry DIPA (0.54 mol) was dissolved in l .2 L dry THF under argon atmosphère and

51.2 mL BuLi solution (0.512 mol, 10M in hexanes) was added dropwise at -78 °C. The mixture was stirred for 45 minutes at the same température. Then 178 g (4-bromo-2chloro-phenoxy)-triisopropyl-silane (0.488 mol) was added dropwise at -78 °C and the white suspension was stirred until no further conversion was observed. Then 36.5 mL Mel (0.586 mmol) was added at this température and the reaction mixture was stirred ovemight without further cooling. The volatiles were evaporated under reduced pressure. The residue was dissolved in 1.5 L EtOAc, washed with brine. The organic phase was dried over Na₂SO₄, filtered and the filtrate was concentrated under reduced pressure. The crude product was filtered through a short pad of silica using hexane as eluent and concentrated under reduced pressure to obtain the product as pale yellow oil. 'H NMR (400 MHz. CDCI3) δ: 7.30 (d. I H), 6.68 (d, l H), 2.53 (s, 3H), l .32 (septet, 3H), l. 14 (d, 18H)

Step C: Préparation B3

178 g (4-broino-2-chloro-3-methyl-phenoxy)-lriisopropyl-silane (0.472 mol) was dissolved in 1.4 L dry THF under argon atmosphère and 52 mL ⁿBuLi solution (0.52 mol, 10M in hexanes) was added dropwise at -78 °C. The mixture was stirred for 5 minutes at this température. Then 116 mL 2-isopropoxy-4,4,5,5-tetramethyl-l ,3,2-dioxaborolane (0.569 mol) was added and the mixture was allowed to warm up to r.t. The volatiles were evaporated under reduced pressure. The residue was dissolved in l .5 L EtOAc, washed with brine. The organic phase was dried over Na₂SO4, filtered and the filtrate was concentrated under reduced pressure. The 2-isopropoxy-4,4,5,5-tetramethyl-1,3,2dioxaborolane impurity was removed by distillation (80 °C at 0.01 mmHg). The crude product was triturated in MeOH affording Préparation B3 as a white solid. *H NMR

ORIGINAL

-68(400 MHz, CDCl₃) δ: 7.53 (d, lH), 6.74 (d. I H), 2.60 (s, 3H), l .34 (s, 12H), l .32 (m, 3H),

I.l2 (d, 18H)

Préparation B4: l-[2-[2-Chloro-3-methyI-4-(4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2yl)phenoxy]cthyl]-4-methyl-piperazine

10.0 g Préparation B2 (37.2 mmol), 8.7 g 2-(4-methylpiperazin-l-yl)ethanol (60.3 mmol) and 15.8 g PPh₃ (60.3 mmol) were dissolved in 100 mL dry toluène and then 27 mL diethyl azodicarboxylate (60.3 mmol, 40 % solution in toluene) was added dropwise. The mixture was stirred at 50 °C under argon until no further conversion was observed. The volatiles were evaporated under reduced pressure and 100 mL Et₂O was added. The precipitated white crystals were filtered off and washed with Et₂O. The filtrate was concentrated under reduced pressure and purified via flash chromatography using CHCl₃and MeOH as eluents. The resulting light brown oil was crystallized from hexane to give Préparation B4 as an off-white solid. 'H NMR (500 MHz. DMSO-dO δ: 7.56 (d, IH),

6.99 (d, IH), 4.15 (t, 2H), 2.72 (t, 2H), 2.51 (s, 3H), 2.50 (br s, 4H), 2.29 (br s, 4H), 2.13 (s, 3H), 1.29 (s, 12H)

Préparation B5: l-[2-[2-Chloro-3-methyl-4-(4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2yl)phenoxy]ethyl]-4-methyl-piperazine

10.0 g Préparation B2 (37.2 mmol), 5.366 g iV./V-dimethylethanolamine (60.3 mmol) and

15.8 g PPh₃ (60.3 mmol) were dissolved in 100 mL dry toluene and then 27 mL diethyl azodicarboxylate (60.3 mmol, 40 % solution in toluene) was added dropwise. The mixture was stirred at 50 °C under argon until no further conversion was observed. The volatiles were evaporated under reduced pressure and 100 mL Et₂O was added. The precipitated white crystals were filtered off and washed with Et₂O. The filtrate was concentrated under reduced pressure and purified via flash chromatography using CHCl₃ and MeOH as eluents. The resulting light brown oil was crystallized from hexane to give Préparation B5. ‘H NMR (400 MHz, DMSO-d₆) δ: 7.56 (d, l H), 6.99 (d, IH), 4.13 (t, 2H), 2.66 (t, 2H), 2.51 (s, 3 H), 2.23 (s, 6H), L29 (s, 12H)

Préparation B6: |4-(5,5-dimethyl-l,3,2-dioxaborinan-2-yl)-3-inethyl-phenoxy|triisopropyl-silane

ORIGINAL

-69Step A: 4-(5,5-dimelhyl-l,3,2-dioxaborinan-2-yl)-3-methyl-phenol

4-675 g (4-hydroxy-2-methyl-phenyl)boronic acid (30.76 mmol), 3.204 neopentyl glycol (32.9 mmol), Amberlyst 15H⁺ and 150 mL 2-Me-THF were stirred at r.t. under N₂atmosphère until no further conversion was observed. The mixture was then filtered through Celite and the filtrate was concentrated under reduced pressure to obtain 4-(5,5-dimethyl-l,3,2-dioxaborinan-2-yl)-3-methyl-phenol. *H NMR (400 MHz. CDCI₃) δ: 7.64 (m, IH), 6.60 (m, 2H), 5.23 (br s, IH), 3.75 (s, 4H), 2.47 (s, 3H), LOI (s, 6H)

Step B: Préparation B6

30.76 mmol 4-(5,5-dimethyl-l,3,2-dioxaborinan-2-yl)-3-methyl-phenol, 8.56 mL TIPSCl (40 mmol) and 4.19 g imidazole (61.52 mmol) were dissolved in 100 mL DCM and stirred at r.t. under N₂ atmosphère until no further conversion was observed. Imidazolium hydrochloride was removed by filtration, the filtrate was concentrated under reduced pressure and purified via flash chromatography using heptane and EtOAc as eluents to obtain Préparation B6. ‘H NMR (400 MHz. CDCI3) δ: 7.62 (d, IH), 6.68-6.66 (m. 2H),

3.76 (s, 4H), 2.47 (s, 3H), 1.32-1.21 (m, 3H), 1.11 (d, 18H), 1.03 (s, 6H)

Préparation B7: 2-(3-bromo-2-methyl-phenyl)-4,4,5,5-tetramethyl-1,3,2dioxaborolane

2.362 g 2,6-dibromo-toluene (9.45 mmol) was dissolved in 10 mL dry THF under N₂atmosphère and the mixture was cooled to -78 °C. Then 5.2 mL nBuLi (2.0M in pentane, 10.4 mmol) was added dropwise and the mixture was stirred for 15 minutes. Then 2.31 mL 2-isopropoxy-4,4,5,5-tetramethyl-l,3,2-dioxaborolane (11.3 mmol) was added dropwise and the mixture was aliowed to warm up to r.t.. It was stirred until no further conversion was observed. Then the mixture was quenched with aqueous NH4CI solution, then extracted with EtOAc. The combined organic layer was dried over Na₂SÜ4, filtered and the filtrate was concentrated under reduced pressure. The crude product was purified via flash chromatography using heptane and EtOAc as eluents to obtain Préparation B7. *H NMR (400 MHz, CDCI3) δ: 7.67 (d, 1 H), 7.62 (d, 1 H), 7.10 (t, 1 H), 2.53 (s, 3H), 1.29 (s, 12H)

Préparation Cl: (2-ethoxypyrimidin-4-yl)methanol

ORIGINAL

-70Step A: 4-(dimethoxymeihyl)-2-ethoxy-pyritnidine

1500 mg Préparation 8b (6.46 mmol) was dissolved in 60 mL éthanol, then 527 mg sodium ethoxide (7.75 mmol) was added and the mixture was stirred at r.t. for l hour. The volatiles were evaporated under reduced pressure and the residue was purified via flash chromatography using heptane and EtOAc as eluents to give 4-(dimethoxymcihyl)-2ethoxy-pyrimidine. MS (M+H); 199.2

Step B: Préparation Cl

Using General Procedure Va and 4-(dimethoxymethyl)-2-ethoxy-pyrimidine as the appropriate acetaL Préparation Cl was obtained. MS (M+H): 155.2

Préparation C2: (l-Butyl-l/f-pyrazol-5-yl)methanol

Using General Procedure Vb and l-butylpyrazole as the appropriate alkyl pyrazole, Préparation C2 was obtained.

'H NMR (400 MHz, DMSO-d₆) δ: 7.30 (d, IH), 6.12 (d, IH), 5.23 (t, IH), 4.49 (d, 2H), 4.06 (t, 2H), 1.72 (m, 2H), 1.26 (m, 2H), 0.88 (t, 3H)

MS (M+H): 155.2

Préparation C3: |2-(2-methoxyphenyl)pyrimidin-4-yl]methanol

Step A : 4-(dimethoxymethyl)-2-(2-methoxyphenyl)pyrimidine

Using General Procedure Vc and 2-methoxybenzamidine acetic acid sait as the appropriate amidine sait, 4-(dimethoxymethyl)-2-(2-methoxyphenyl)pyrimidine was obtained. 'H NMR (400 MHz, DMSO-d_ô) 6: 8.93 (d, IH), 7.55-7.44 (m, 3H), 7.16 (d, IH), 7.06 (m, IH), 5.31 (s, IH), 3.76 (s, 3H), 3.37 (s, 6H)

Step B: Préparation C3

261 mg 4-(dimethoxymethyl)-2-(2-methoxyphenyr)pyrimidine (l.O mmol) was dissolved in 2 mL HCl in dioxane (4M solution), then 2 mL water was added and this mixture was stirred at 50 °C for 16 hours. The reaction mixture was cooled to 0 ^DC, then 320 mg NaOH (8.0 mmol) was added portionwise. The pH was adjusted to 8 using 10 % K2CO3 solution,

ORIGINAL

-7l · then 76 mg sodium borohydride (2.0 mmol) was added and the mixture was stirred for 30 minutes at 0 °C. The reaction mixture was diluted with 5 mL water and extracted with EtOAc. The combined organic phases were dried over Na₂SO₄. filtered and the filtrate was concentrated under reduced pressure. The crude product was purified via flash 5 chromatography using heptane and EtOAc as eluents to give Préparation C3. ’H NMR (400 MHz, DMSO-dû) δ: 8.84 (d, IH), 7.50-7.42 (m. 3H), 7.14 (d, IH), 7.03 (m, IH), 5.66 (t, lH),4.58(d, 2H), 3.75 (s, 3H)

Préparation C4: (I-/c/7-But\l-l7/-pyrazol-5-yI)methanoI

Step A: I-\exl-butyl-5-(dimethoxymethyl)-l\A-pyrazole

Using General Procedure Vd and /e/7-butylhydrazine hydrochloride as the appropriate hydrazine hydrochloride, l-/er/-butyl-5-(dimethoxymethyl)-lf/-pyrazole was obtained. *H NMR (400 MHz, DMSO-d₆) ô: 7.34 (d, IH), 6.34 (d, IH), 5.74 (s, IH), 3.24 (s, 6H), L57 (s, 9H)

Noie: l-/e/7-butyl-3-(dimethoxymethyl)-l/f-pyrazole was also obtained. ’H NMR (400 I5 MHz, DMSO-d₆) Ô: 7.75 (d, IH), 6.18 (d, IH), 5.34 (s, IH), 3.24 (s, 6H), 1.50 (s, 9H)

Step B: Préparation C4

Using General Procedure Ve and l-/e/7-butyl-5-(dimethoxymethyl)-l/7-pyrazole as the appropriate acetal, Préparation C4 was obtained.'H NMR (400 MHz, DMSO-dû) δ: 7.27 (d, IH), 6.19 (d, IH), 5.31 (t, 1 H), 4.61 (d, 2H), 1.56 (s, 9H)

Préparation C5: [2-(2-methoxyethyl)pyrimidin-4-yl] methanol

Step A : 4-(dhnethoxyniethyl)-2-(2-methoxyethyl)pyrimidine

Using General Procedure Vc and 3-methoxypropanamidine hydrochloride as the appropriate amidîne hydrochloride, 4-(dimethoxyrnethyi)-2-(2-rnethoxyethyl)pyrimidine was obtained. 'H NMR (400 MHz. DMSO-d₆) δ: 8.78 (d, IH). 7.38 (d, IH), 5.25 (s, JH), 25 3.80 (t, 2H), 3.33 (s, 6H), 3.22 (s, 3H), 3.11 (t, 2H)

Note: 2-[4-(dimethoxymethyl)pyrimidin-2-yl]-V,V-dimethyI-ethanamine was also obtained. MS (M+H): 226.2

ORIGINAL

-72Step B: Préparation C5

Using General Procedure Va and 4-(dimethoxymethyl)-2-(2-methoxyethyl)pyrimidine as the appropriate acetal, Préparation C5 was obtained. 'H NMR (400 MHz, DMSO-dr,) δ: 8.70 (d, IH), 7.39 (d, lH), 5.60 (t, IH), 4.52 (d, 2H), 3.78 (t, 2H), 3.22 (s, 3H), 3.06 (t, 2H)

Préparation C6: |l-(2,2,2-Triiluoroethyl)-177-pyrazol-5-yljmethanoI

Step A: 5-(dimelhoxymethyl)-l-(2,2,2-trifluoroethyl)-4,5-dihydro-lW-pyrazol-5-ol

Using General Procedure Vd in the absence of sodium methoxide and using éthanol instead of methanoi and 2,2,2-trifluoroethylhydrazine (70 w/w% in water) as the appropriate hydrazine, 5-(dimethoxymethyl)- l -(2,2.2-trifluoroethyl)-4,5-dihydro-1 Hpyrazol-5-ol was obtained. ’H NMR (400 MHz, DMSO-d₆) δ: 6.83 (t, IH), 6.03 (s, IH),

4.30 (s, IH), 3.95 (m, IH), 3.47 (m, IH), 3.40 (d, 6H), 2.88 (m. IH), 2.50 (m, IH)

Step B: Préparation C6

Using General Procedure Ve and 5-(dimethoxymethyI)-1-(2,2,2-trifluoroethyl)-4,5dihydro-17f-pyrazol-5-ol as the appropriate acetal, Préparation C6 was obtained. ^fH NMR (400 MHz. DMSO-dû) δ: 7.48 (d, IH), 6.27 (d, IH), 5.46 (t, IH), 5.08 (q, 2H). 4.56 (d, 2H)

Préparation C7: (2-(Morpholin-4-yl)pyrimidin-4-yl)methanol

Step A: 4-[4-(dimethoxymethyl)pyrimidin-2-yl] morpholine

25.0 g Préparation 8b (107.6 mmol) was dissolved in 161 mL morpholine and the mixture was stirred at r.t. until no further conversion was observed. Then it was concentrated under reduced pressure and the crude product was purified via flash chiomatography using heptane and EtOAc as eluents to obtain 4-[4-(dimethoxymethyl)pyrimidin-2yl]morpholine.

Step B: Préparation C7

ORIGINAL

-73Using General Procedure Va and 4-[4-(dimethoxymethyl)pyrimidin-2-yl]morpholine as the appropriate acetal, Préparation C7 was obtained. NMR (400 MHz, DMSO-dr,) δ: 8.35 (d, IH), 6.75 (dm, IH), 5.431 (t, IH), 4.36 (dm, 2H), 3.67 (m, 4H), 3.63 (m, 4H)

Préparation C8: [2-(2,2,2-trifluoroethoxy)pyrimidin-4-yl]methanol

Step A: 4-(dimethoxymelhyl)-2-(2,2,2-trifluoroethoxy)pyrimidine

5.00 g Préparation 8b (21.5 mmol) was dissolved in 54 mL dry acetonitrile, then 5.95 g K.7CO3 (43J mmol) and 3.24 g 2,2,2-trifluoroethanol (32.3 mmol) were added, and the mixture was stirred at 60 °C until no further conversion was observed. The reaction mixture was cooled, filtered, the solid was washed with EtOAc, then the filtrate was concentrated under reduced pressure. The crude product was purified via flash chromatography using heptane and EtOAc as eluents to give 4-(dimethoxymethyl)-2(2,2,2-trifluoroethoxy) pyrimidine. ’H NMR (400 MHz. DMSO-d₆) δ: 8.74 (d, IH), 7.32 (d, IH), 5.25 (s, IH), 5.05 (q, 2H), 3.34 (s, 6H)

Step B: Préparation C8

Using General Procedure Va and 4-(dimethoxymethyl)-2-(2,2,2-trifluoroethoxy) pyrimidine as the appropriate acetal, Préparation C8 was obtained. ¹H NMR (400 MHz, DMSO-d(,) ô: 8.65 (d, IH), 7.32 (d, IH), 5.69 (t. 1 H), 5.02 (q, 2H), 4.51 (d, 2H)

Préparation C9: |2-(2-FluorophenyI)pyrimidin-4-yl| methanol

Step A: 2-Fiuoro-N'-hydroxy-benzamidine

The mixture of 11.48 g hydroxylamine hydrochloride (165 mmol), 13.87 g NaHCO3 (165 mmol) and 120 mL MeOH was stirred at r.t. for 30 minutes. Then 10 g 2-fluorobenzonitrile (82.6 mmol) was added and the mixture was stirred at 75 °C until no further conversion was observed. The solvent was partially evaporated under reduced pressure, the residue was filtered, washed with MeOH. The filtrate was concentrated under reduced pressure, then diluted with water and extracted with EtOAc. The combined organic phases were dried over Na2SO₄, filtered and the filtrate was concentrated under reduced pressure to give 2-fluoro-7V-hydroxy-benzamidine.

ORIGINAL

-74Step B : 2-Fluoro benzamidine

12.67 g 2-fluoro-A-hydroxy-benzamidine (81.55 mmol) was dissolved in 300 mL AcOH at 0 °C and 9.24 mL AC2O (97.86 mmol) was added. The mixture was stirred at r.t. until no further conversion was observed. Then 630 mg 10 % Pd/C was added and the mixture was 5 stirred under H2 atmosphère (4 bars) until no further conversion was observed. The mixture was filtered through celite and the filtrate was concentrated under reduced pressure to obtain 2-fiuorobenzamidine acetate. MS (M(free base)+H): 139.4

Step C: 4-(Dimethoxymethyl)-2-(2-flnorophenyl)pyrimidine

Using General Procedure Vc and 2-fluorobenzamidine as the appropriate amidine, I0 4-(dimethoxymethyl)-2-(2-fluorophenyl)pyrimidine was obtained. MS (M+H): 249.2

Step D: Préparation C9

Using General Procedure Va and 4-(dimethoxymethyl)-2-(2-fluorophenyl)pyrimidine as the appropriate acetal, Préparation C9 was obtained. MS (M+H): 205.2

Préparation CIO: [2-|2-(2-methoxyethoxy)phcnyl|pyrimidin-4-yl|methanoI

Step A : N ’-Hydroxy-2-methoxyethoxy-benzamidine eq. hydroxylamine hydrochloride was dissolved in MeOH (ImL/mmol), then 2 eq. NaHCOj was added. The mixture was stirred at r.t. for 20 minutes, then l eq. 2-methoxyethoxy-benzonitrile was added and the mixture was stirred at reflux until no further conversion was observed. MeOH was partially evaporated, the residue was filtered 20 and the filtrate was concentrated under reduced pressure. The obtained A-hydroxy-2-(2methoxyethoxy)benzamidine was used without further purification.

'H NMR (400 MHz. CDCI3) Ô: 9.48 (s, IH), 7.45 (m, l H), 7.34 (m, IH). 7.08 (d, IH), 6.94 (td, IH), 5.65 (br s, 2H), 4.17 (m, 2H), 3.67 (m, 2H), 3.31 (s, 3H)

MS (M+H): 2H.2

Step B: 2-methoxyethoxy-benzamidine

ORIGINAL

8.22 g ÀMiydroxy-2-(2-methoxyethoxy)benzamidine (39.1 mmol) was dissoived in 80 mL AcOH at 0 °C, then 4.43 mL Ac₂O (46.92 mmol) was added dropwise. The mixture was stirred at r.t. until no further conversion was observed. 575 mg 10 % Pd/C was added and the mixture was stirred under H₂ atmosphère (4 bars) until no further conversion was 5 observed. The mixture was filtered through ceiite and the filtrate was concentrated under reduced pressure to obtain 2-(2-methoxyethoxy)benzamidine acetate. MS (M+H): 195.2

Siep C: 4-(Dimethoxymethyl)-2f2~methoxyethoxy-phenyl]pyrimidine

Using General procedure Vc and 2-(2-methoxyethoxy)benzamidine acetate as the appropriate amidine sait, 4-(dimethoxymethyl)-2-[2-methoxyethoxy-phenyl]pyrimidine 10 was obtained.

'H NMR (400 MHz, CDCl₃) δ: 8.92 (d, IH), 7.55 (m, IH), 7.47 (m. IH), 7.45 (m, IH),

7.17 (d, IH), 7.08 (m, IH), 5.29 (s, IH), 4.12 (m, 2H), 3.57 (m, 2H), 3.36 (s, 6H), 3.20 (s, 3H)

MS (M+H): 305.0

Step D: Préparation CIO

Using General Procedure Va and 4-(dimethoxymethyl)-2-[2-methoxyethoxyphenyl]pyrimîdine as the appropriate acetal. Préparation CIO was obtained.

'H NMR (400 MHz. CDCl₃) δ: 8.84 (d, IH), 7.53 (m, IH), 7.47 (m, IH), 7.43 (m. IH),

7.14 (d, IH), 7.05 (td, IH), 5.64 (t, IH), 4.58 (d, 2H), 4.11 (m, 2H), 3.57 (m. 2H), 3.21 (s, 20 3 H)

MS (M+H): 261.0

Example 1: jV-|(5S„)-5-{3-chIoro-2-methyl-4-|2-(4-niethylpiperazÎn-l-yl)ethoxy] phenyI}-6-(4-fluorophenyl)thieno[2,3-(/]pyrimidin-4-yl|-2-methoxy-Z)-phenylalanine and

Example 2: jV-[(57?_a)-5-{3-chloro-2-niethyI-4-|2-(4-methylpiperazin-l -yl)ethoxy | phenyl}-6-(4-fluorophenyl)thieno[2,3-rf| pyrimidin-4-yl|-2-methoxy-D-phenyIalanine

ORIGINAL

-76Using General Procedure VI and Préparation 7a as the appropriate phénol dérivative and methanol as the appropriate alcohol, then hydrolyzing the formed intermediate according to General Procedure Vil, Exampie 1 was obtained as the earlier eluting diastereoisomer. HRMS calculated for C36H37CIFN5O4S: 689.2240, found: 345.6182 (M+2H)

Example 2 was obtained as the later eluting diastereoisomer. HRMS calculated for C36H37CIFN5O4S: 689.2240, found: 345.6185 (M+2H)

Example 3: jV-|(55_(J)-5-{3-chloro-2-methyl-4-[2“(4-methylpiperazin-l-yl)ethoxy| phenyl}-6-(4-nuorophenyl)thieno[2,3-rf]pyrimidin-4-yl]-2-[(l-methyl-lFf-pyrazol-5-yl) methoxy|-Z)-phenylalanine

Using General Procedure VI and Préparation 7ad2 as the appropriate phénol dérivative and (2-methylpyrazol-3-yl)methanol as the appropriate alcohol, then hydrolyzing the formed intermediate according to General Procedure VU, Example 3 was obtained. HRMS calculated for C4oH₄₁ClFN₇0₄S: 769.2613, found: 385.6378 (M+2H)

Exampie 4: 7V-[(55„)-5-{3-chloro-2-methyl-4-|2-(4-methylpiperazin-l-yI)ethoxy] phenyl}-6-(4-fluorophenyl)thieno[2,3-rf]pyrimidin-4-yl]-2-[(2-ethoxypyrimidin-4-yI) methoxy]-Z)-phenylalanine and

Example 5: A^r-[(57f_a)-5-{3~chloro-2-methyl-4-[2-(4-methylpiperazin-l-yl)ethoxy| phenyl}-6-(4-fIuorophenyl)thicno[2,3“rf]pyrimidin-4-yl]-2-[(2-ethoxypyrimidin-4-yI) methoxyj-Z>-phenylalanine

Using General Procedure VI and Préparation 7a as lhe appropriate phénol dérivative and Préparation Cl as the appropriate alcohol, then hydrolyzing the formed intermediate according to General Procedure VII, a mixture of dîaslereoisomers was obtained. They were separated via préparative reversed phase chromatography using 40 mM aqueous NH₄OAc solution (pH = 4, adjusted with AcOH) and acetonitrile as eluents, Example 4 was obtained as the earlier eluting diastereoisomer. HRMS calculated for C42H43CIFN7O5S: 811.2719, found: 406.6417 (M+2H)

ORIGINAL

-77Example 5 was obtained as the later eluting diastereoisomer. HRMS calculated for C42H43CIFN7O5S: 811.2719, found: 406.6436 (M+2H)

Example 6: 2-|(l-butyi-l/7-pyrazol-5-yl)methoxy|-7V-[(5_>S^, _i,)-5-{3-chloro-2-methyl-4-|2(4-methylpïperazin-l-yI)ethoxy]phenyl}-6-(4-fIuorophenyl)thieno[2,3-rf]pyrimidin-4yl]-Z>-phenylalanine

Using General Procedure VI and Préparation 7ad2 as the appropriate phénol dérivative and Préparation C2 as the appropriate alcohol, then hydrolyzing the formed intermediate according to General Procedure VII, Example 6 was obtained. HRMS calculated for C43H47CIFN7O4S: 811.3082, found: 406.6616 (M+2H)

Example 7: A-|(5.S’,,)-5-{3-chl()r()-2-nietliyl-4-|2-(4-inethvlpiperazin-l-yl)etlioxy] phenyl}-6-(4-fluorophenyl)thieno[2,3-i/]pyrimidin-4-yl]-2-{[2-(2-methoxyphenyI) pyrimidin-4-yl]methoxy)-Z)-phenylalanine

Using General Procedure VI and Préparation 7ad2 as the appropriate phénol dérivative and Préparation C3 as the appropriate alcohol, then hydrolyzing the formed intermediate according to General Procedure VII, Example 7 was obtained. HRMS calculated for C47H45CIFN7O5S: 873.2875, found: 437.6498 (M+2H)

Example 8: jV-[(5S_a)-5-{3-chioro-2-methyl-4-[2-(4-methylpiperazin-l-yl)ethoxy] phenyl}-6-(furan-2-yl)thieno|2,3-iZ]pyrimidin-4-yl|-2-metlioxy-Z)-phenylalanine and

Example 9: jV-KS/fiJ-â-p-chloro^-methylT-^-^-mcthylpipcrazin-l-yOetlioxyl phenyl}-6-(furan-2-yl)thieno[2,3-i7]pyrimidin~4-yl|-2-inethoxy-Z)-phcnylalanine

Using General Procedure le and Préparation 4b as the appropriate 4-chloro-lhicno|2,3-i/| pyrimidine dérivative and (2Â)-2-amino-3-(2-methoxyphenyl)propanoic acid as the appropriate amino acid dérivative, Example 8 was obtained as the earlier eluting diastereoisomer. HRMS calculated for C34H36CIN5O5S: 661.2126, found: 662.2203 (M+H)

ORIGINAL

-78Example 9 was obtained as the later eluting diastereoisomer. HRMS calculated for C34H36CIN5O5S: 661.2126,, found: 662.2203 (M+H)

Example 10: 2-chloro-/V-|5-{3-chloro-2-methyl-4-|2-(4-methylpiperazin-l-yl)cthoxy| phenyl}-6-(furan-2-yl)thieno[2,3-iZ]pyrimidin-4-yl]-Z)-phenylaianine, diastereoisomer 1

Using General Procedure le and Préparation 4b as the appropriate 4-chloro-thieno[2,3-i/] pyrimidine derivative and (2Æ)-2-amino-3-(2-chlorophenyl)propanoic acid as the appropriate amino acid derivative, Example 10 was obtained as the earlier eluting diastereoisomer. HRMS calculated for C33H33CI2N5O4S: 665.1630, found: 666.1670 10 (M+H)

Example 11: 2-carbamoyl-jV-[5-{3-chloro-2-niethyl-4-[2-(4-methylpiperazin-l-yi) ethoxy]phcnyl}-6-(furan-2-yl)thieno|2,3-(7|pyrimidin-4-yl]-Z>-phenylalanine

Step A: 5-[3-chloro-2-methyl-4-[2-(4-methylpiperazin-l-yl)elhoxy]phenyl]-4-fluoro-6-(2furyl)thieno[2.3-à]pyrimidine

A mixture of 150 mg Préparation 4b (0.3 mmol) and 380 mg silver fluoride (3.0 mmol) in 6 mL toluene was heated at reflux température for 3 hours. Then it was cooled to r.t., and the inorganic components were fîitered off. The filtrate was concentrated under reduced pressure to obtain the crude product which was used in the next step without further -J purification.

Step B: Example II

A mixture of 316 mg 5-[3-chIoro-2-methyl-4-[2-(4-methylpiperazin-l-yl)ethoxy]phenyl]4-fluoro-6-(2-furyl)thieno[2,3-iZ]pyrimidine (0.65 mmol), 271 mg (27?)-2-amino-3-(2carbamoylphenyl)propanoic acid (1.30 mmol) and 424 mg Cs₂CO₃ (1.30 mmol) in 6 mL DMSO was stirred at 40 °C for 30 minutes. The mixture was dïluted with water, the pH 25 was adjusted to 5 using 1M HCl solution and extracted with DCM. The organic phase was dried over Na₂SO4, fîitered and the filtrate was concentrated under reduced pressure. The crude product was purified via préparative reversed phase chromatography using 0.1 %

ORIGINAL

-79aqueous TFA solution solution and acetonitrile as eluents. The diastereoisomer eiuting © earlier was collected as Example 11. HRMS calculated for C₃4H₃₅C1N₆O₅S: 674.2078, found: 675.2146 (M+H)

Example 12: A^r-[(51S^ra)-5-{3-chIoro-2-methyl-4-[2-(4-methylpiperazin-l-yl)ethoxy| 5 phenyl}-6-(furan-2-yl)thieno[2,3-i/|pyrimidin-4-yl|-2-(pyridin-2-ylmethoxy)-£>phenylalanine and

Example 13: /V-[(57î„)-5-{3-chloro-2-methyI-4-|2-(4-methylpiperazin-l-yl)ethoxy] phenyl}-6-(furan-2-yl)thieno[2,3-i7] pyrimidin-4-yl]-2-(pyridin-2-ylmetlioxy)-Z>10 phenylalanine

Using General Procedure le and Préparation 4b as the appropriate 4-chloro-thieno [2,3-/7] pyrimidine dérivative and Préparation A5 as the appropriate amino acid dérivative. Example 12 was obtained as the later eiuting diastereoisomer. HRMS calculated for C₃₉H₃₉C1N₆O₅S: 738.2391, found: 370.1269 (M+2H)

Example 13 was obtained as the earlier eiuting diastereoisomer. HRMS calculated for C^H-uClN^OsS: 738.2391, found: 370,1263 (M+2H)

Example 14: /V-[(5S_fl)-5-{3-chloro-2-methyl-4-[2-(4-methylpiperazin-l-yl)ethoxy] phenyl}-6-(5-fluorofuran-2-yl)thieno|2,3-iZ]pyriniidin-4-yl]-2-hydroxy-Z>phenylalanine

Step A: (2R)-2-[[(5S_a)-5-[3-chloro-2-nielhyl-4-[2-(4-melhylpiperazin-l-yl)ethoxy]phenylJ6-iodo-thieno[2,3-d]pyrimidin-4-yl]amino]-3-(2-hydroxyphenyl)propanoic acid

Using General Procedure le and Préparation 4a as the appropriate 4-chloro-thieno[2,3-/7] pyrimidine dérivative and (2/î)-2-amino-3-(2-hydroxyphenyl)propanoic acid a mixture of diastereoisomers was obtained. They were separated via HILIC chromatography.

(27?)-2-[[(5S'_o)-5-[3-chloro-2-methyl-4-[2-(4-inethylpiperazin-l-yl)ethoxy]phenyl]-6-iodothieno[2,3-/7]pyrimidin-4-yl]amino]-3-(2-hydroxyphenyl)propanoic acid was obtained as the later eiuting diastereoisomer. MS (M+H): 708.0

ORIGINAL

-80Step B: Example 14

Using General procedure IHb and (2Æ)-2-[[(5Sj-5-[3-chloro-2-mcthyl-4-[2-(4-methyl piperazin-I-yl)ethoxy]phenyl]-6-iodo-thieno[2,3-6/]pyrimidin-4-yl]amino]-3-(2-hydroxy phenyl)propanoic acid as the appropriate 6-iodo-thieno[2,3</]pyrimidine dérivative and 2-(5-fluoro-2-furyl)-4,4,5,5-tetramethyl-l,3,2-dioxaborolane as the appropriate boronic ester dérivative. Example 14 was obtained. HRMS calculated for C33H33CIFN5O5S: 665.1875, found: 333.6012 (M+2H)

Example 15: /V-[(5/G)-5-{3-chloro-2-methyl-4-[2-(4-methylpiperazin-I-yl)ethoxy] phenyl}-6-(5-fluorofuran-2-yl)thieno[2,3-<7| pyrimidin-4-yl]-2-methoxy-Z>phenylalanine and

Example 16: A’-[(5A\_i)-5-{3-chloro-2-nicthyl-4-[2-(4-methylpipcrazin-l-yl)ethoxy] phenyl}-6-(5-fiuorofuran-2-yl)thieno|2,3-J]pyrimidin-4-yl|-2-mcthoxy-Z>phenylalanine

Using General Procedure Ib and Préparation 4c as the appropriate 4-chloro-thieno[2.3-<7] pyrimidine dérivative and (27?)-2-amino-3-(2-methoxyphenyl)propanoic acid as the appropriate amino acid dérivative. Example 15 was obtained as the later eluting diastereoisomer. HRMS calculated for C34H35CIFN5O5S: 679.2031, found: 680.2100 (M+H)

Example 16 was obtained as the earlier eluting diastereoisomer. HRMS calculated for C34H35CIFN5O5S: 679.2031, found: 680.2092 (M+H)

Example 17: 7V-|(5S„)-5-{3-chloro-2-methyl-4-[2-(4-methylpiperazin-l-yi)etlioxy| phenyl}-6-(5-fluorofuran-2-yl)thieno|2,3-rflpyrimidin-4-yl|-2-(2,2,2-trifluoroethoxy)Z>-phenyIalanine

Step A: ethyl (2R.)-2-[[(5S^-5-[3-chloro-2-methyl-4-[2-(4-methylpiperazin-l-yl)ethoxy] phenyl]-6-iodo-thieno[2,3-à]pyrimidin-4-yl]amino]-3-(2-hydroxyphenyl)propanoate

876 mg (2Æ)-2-[[(5S_a)-5-[3-chloro-2-methyl-4-[2-(4-methylpiperazin-l-yl)ethoxy]phenyl]6-iodo-thieno[2,3-i/]pyrimidin-4-yI]amino]-3-(2-hydroxyphenyl)propanoic acid

ORIGINAL

-8l (1.24 mmol) was dissolved in 5 mL éthanol then 0.05 mL concentrated sulfuric acid was added and the mixture was stirred at 70 °C for 2 hours. Then the mixture was diluted with water, the pH was set to 5 using IM NaHCCh solution and extracted with DCM. The organic phase was dried over Na₂SO4, filtrated and the filtrate was concentrated under reduced pressure. The crude product was purified via flash chromatography using DCM and MeOH as eluents to obtain ethyl (2/?)-2-[[(5SJ-5-[3-chloro-2-methyl-4-[2-(4methylpiperazin-l-yl)ethoxy]phenyl]-6-iodo-thieno[2,3-i/]pyrimidin-4-yl]amino]-3-(2hydroxyphenyl) propanoate. MS (M+H): 736.1

Step B: ethyl (2VÇ-2-[[(5§^-5-[3-chloro-2-methyî-4-[2-(4-meihylpiperazm-l-yl)ethoxy] phenyl]-6-iodo-thieno[2.3-d]pyrûnidin-4-yl]amino]-3-[2-(2₁2,2-lrifluoroethoxy)phenyl] propanoate 648 mg ethyl (27?)-2-[[(55_a)-5-[3-chloro-2-methyl-4-[2-(4-inethylpiperazin-l-yl)ethoxy] phenyl]-6-iodo-thieno[2.3-i/]pyrimidin-4-yl]amino]-3-(2-hydroxyphenyl)propanoate (0.88 mmol) was dissolved in 10 mL DMF then 415 mg K₂CO₂ (3.00 mmol) and 348 mg

2,2,2-trifluoroethyl trifluoromethanesuilbnate (1.50 mmol) were added at r.t. The mixture was stirred at 50 °C for 5 hours. The reaction mixture was diluted with brine, extracted with DCM. The organic phase was dried over Na₂SÛ4, filtered and the filtrate was concentrated under reduced pressure. The crude material was purified via flash chromatography using DCM and methanol as eluents to obtain ethyl (2/f)-2-[[(5S_a)-5-[3chloro-2-niethyl-4-[2“(4-methylpiperazin-l-yl)ethoxy]phenyl]-6-iodo-thieno[2,3-<7] pyriinidin-4-yl]arnino]-3-[2-(2,2,2-trifluoiOethoxy)phenyl]propanoate. MS (M+H); 818.1

Step C: Example 17

Ethyl (2Æ)-2-[[(5S'_a)-5-[3-chloro-2-methyl-4-[2-(4-methylpiperazin-l-yl)ethoxy]phenyl]-6iodo-thieno[2,3-<7]pyrimidin-4-yl]amino]-3-[2-(2,2,2-trifluoroethoxy)phenyI]propanoate was hydrolyzed according to General procedure VII to give (2Æ)-2-[[(5.S'_a)-5-[3-chloro-2methyl-4-[2-(4-methylpiperazin-l-yl)ethoxy]phenyl]-6-iodo-thieno[2,3-i/]pyrimidin-4-yl] amino]-3-[2-(2,2,2-trifluoroethoxy)phenyl]propanoic acid. This compound was used as the appropriate 6-iodo-thieno[2,3-i/]pyrimidine dérivative and converted to Example 17 according to General Procedure Hlb, using 2-(5-fluoro-2-furyl)-4,4,5,5-tetramethyl-l,3,2ORIGINAL

-82dioxaborolane as the appropriate boronic acid dérivative. HRMS calculated for C35H34CIF4N5O5S: 747.1905, found: 374.6006 (M+2H)

Example 18: A'-|(5/?„)-5-{3-chloro-2-nu‘thyl-4-[2-(4-inethylpipcrazin-l-yl)ethoxy| phenyl}-6-(5-fluorofuran-2-yl)thieno [2,3-^1 pyrimidin-4-yl|-2-(pyridin-2-ylmcthoxy)£)-phenylalanine and

Example 19: A^r-[(5S^, _fl)-5-{3-chIoro-2-methyl-4-[2-(4-methylpiperazin-l-yl)ethoxyI phenyl}-6-(5-fluorofuran-2-yl)thieno|2,3-rf]pyrimidin-4-yl]-2-(pyridin-2-ylniethoxy)D-phenylalanine

Using General Procedure ic and Préparation 4c as the appropriate 4-chloro-thieno[2,3-c/J pyrimidine dérivative and Préparation A5 as the appropriate amino acid dérivative, Example 18 was obtained as the earlier eluting diastereoisomer. HRMS calculated for C₃₉H3_SC1FN₆O₅S: 756.2296, found: 379.1230 (M+2H)

Example 19 was obtained as the later eluting diastereoisomer. HRMS calculated for C39H38CIFN6O5S: 756.2296, found: 379.1230 (M+2H)

Example 20: _JV-|(55„)-5-{3-chloro-2-methyl-4-|2-(4-methyIpiperazin-l-yl)ethoxy] phenyl}-6-(5-fluorofuran-2-yl)thieno[2,3-i/] pyrimidin-4-yl]-2-|(l-mcthyl-17/-pyrazol5-yl)methoxy]-Z)-phenylalanine

Using General Procedure II 1b and Préparation 5b as the appropriate 6-iodo-thieno[2,3-i/] pyrimidine dérivative and 2-(5-fluoro-2-furyl)-4,4,5,5-tetramethyl-l,3,2-dioxaborolane as the appropriate boronic acid dérivative, Example 20 was obtained. HRMS calculated for C3₈H3₉C1FN7O₅S: 759.2406, found: 380.6271 (M+2H)

Example 21: 7V-[(5S„)-5-{3Chloro-2-methyl-4-|2-(4-methylpiperazin-l-yl)ethoxyl phenyI}-6-(5-fluorofuran-2-yl)thieno|2,3-i7]pyrÎmidÎn-4-yl|-2-[(l-ethyl-l/7-pyrazol-5yl)methoxy]-D-phenylalanine

ORIGINAL

- 83 Using General Procedure Illb and Préparation 5c as the appropriate 6-iodo-thieno[2,3-cf| pyrimidine dérivative and 2-(5-fluoro-2-furyl)-4,4,5,5-tetramethy!-l,3,2-dioxaborolane as the appropriate boronic acid dérivative. Example 21 was obtained. HRMS calculated for C39H41CIFN7O5S: 773.2562, found: 387.6358 (M+2H)

Example 22: 7V-[(55₀)-5-{3-chloro-2-methyl-4-|2(4-methyIpiperazin-l-yl)ethoxy] phenyl}-6-(5-fluorofuran-2-yl)thieno[2,3-rf]pyrimidm-4-yI]-2-|(2-ethoxypyrimidin-4yl)methoxy|-Z)-phenylalanine

Using General Procedure le and Préparation 4c as the appropriate 4-chloro-thieno[2,3-t/] pyrimidine dérivative and Préparation A2 as the appropriate amino acid dérivative. Example 22 was obtained as the later elutîng diastereoisomer. HRMS calculated for C40H41CIFN7O6S: 801.2512, found: 401.6326 (M+2H)

Example 23: 2-|(l-butyl-l//-pyraz.ol-5-yl)methoxy|-.V-|(5A\)-5-|3-chloro-2-inethyl-4[2-(4-methylpiperazin-l-yl)ethoxy]phenyl}-6-(5-fluorofuran-2-yl)thieno[2,3-rf] pyrimidin-4-yl]-£)-phenylalanine and

Example 24: 2-[(l-butyl-lH-pyrazol-5-yl)methoxy]-7V-[(5S_a)-5-{3-chloro-2-methyl-4j2-(4-methyIpiperazin-l-yl)ethoxy|phenyl}-6-(5-fluorofuran-2-yl)thicno[2,3rf]pyrimidin-4-yl]-£)-phenylalanine

Using General Procedure le and Préparation 4c as the appropriate 4-chloro-thieno[2,3-c/] pyrimidine dérivative and Préparation A3 as the appropriate amino acid dérivative. Example 23 was obtained as the earlier eluting diastereoisomer. HRMS calculated for C41H45CIFN7O5S: 801.2875, found: 401.6502 (M+2H)

Example 24 was obtained as the later eluting diastereoisomer. HRMS calculated for C41H45CIFN7O5S: 801.2875, found: 401.6505 (M+2H)

Example 25: 7V-[(51ï_a)-5-{3-chloro-2-methyl-4-|2-(4-methylpiperazin-l-yl)ethoxy] phenyl}-6-(5-fluorofuran-2-yI)thieno[2,3-r/]pyrimidin-4-yl]-2-{|2-(2,2,2-trifluoro ethoxy)pyrimidin-4-yl]mcthoxy}-I>-phenylalanine

ORIGINAL

-84and

Example 26: N-[(55_tf)-5-{3-chloro-2-methyI-4-[2-(4-methylpiperazin-l-yl)ethoxy] phenyl}-6-(5-fluorofuran-2-yl)thieno[2,3-rf|pyriniidin-4-ylj-2-{|2-(2,2,2-trifluoro ethoxy)pyrinüdin-4-yl]metIu)xy}-D-phenylalanine

Using General Procedure le and Préparation 4c as the appropriate 4-chloro-thieno[2,3-i/] pyrimidine dérivative and Préparation A8 as the appropriate amino acid dérivative, Example 25 was obtained as the earlier eluting diastereoisomer. HRMS calculated for C_4üH3_gClF4N₇O6S: 855.2228, found: 428.6181 (M+2H)

Example 26 was obtained as the later eluting diastereoisomer. HRMS calculated for C₄₀H3_SCIF4N₇O₆S: 855.2228, found: 428.6193 (M+2H)

Example 27: jV-[(55₀)-5-{3-chloro-2-methyl-4-[2-(4-methylpiperazin-l-yl)ethoxy| phenyl}-6-(5-fluorofuran-2-yl)thieno[2,3-i/]pyrimidin-4-yl|-2-{[2-(2-methoxyphenyl) pyrimidin-4-yl]methoxy}-Z)-phenyialanine

Step A: ethyl (2\4)~2-][(5‘ip-5-[3-chlofo-2-methyl-4-[2-(4-methylpipercizin-l-yi)elhoxy] phenyl]-6-(5-fluoro-2-fwyl)thieno[2,3-à]pyrimidin-4-yl]amino]-3-(2-hydroxyphenyl) propanoate

0.97 g Example 14 (1.46 mmol) was dissolved in 15 mL HCl solution (1.25M in EtOH) and stirred at 40 °C overnight. The mixture was cooled to r.t., neutralized with aqueous NaHCO3 solution and the mixture was extracted with DCM. The organic phase was dried over Na₂SO4, filtered and the filtrate was concentrated under reduced pressure. The residue was purified via flash chromatography using DCM and MeOH as eluents to obtain ethyl (2Â)-2-[[(5S_fl)-5-[3-chloro-2-methyl-4-[2-(4-methylpiperazin-l-yl) ethoxy]phenyl]-6-(5fluoro-2-furyl)thieno[2,3-i7)pyrimidin-4-yl]amino]-3-(2-hydroxy phenyl)propanoate.

^lH NMR (400 MHz, DMSO-d₆) δ: 9.48 (br s, IH), 8.39 (s, IH), 7.30 (s, 2H), 7.01 (td, IH),

6.72 (d, IH), 6.64 (t, IH), 6.41 (d, IH), 5.83 (m, IH), 5.56 (t, IH), 5.08 (d, 1 H), 4.94 (m, IH), 4.30 (t, 2H), 4.03 (m, 2H), 3.07 (dd, IH), 2.81 (t, 2H), 2.56 (br, 4H), 2.36 (dd, IH),

2.32 (br, 4H), 2.14 (s, 3H), 1.91 (s, 3H)

MS (M+H): 694.2

ORIGINAL

-85 Step B: Example 27

Using General Procedure VI and ethyl (27?)“2-[[(5S_fl)-5-[3-chloro-2-methyl-4-[2-(4-methyl piperazin-l-yl)ethoxy]phenyl]-6-(5-fluoro-2-furyI)thieno[2,3-ri]pyrimidin-4-yl]amino]-3(2-hydroxyphenyl)propanoate as the appropriate phénol dérivative and Préparation C3 as the appropriate alcohol dérivative, then hydrolyzing the formed intermediate according to General Procedure VII, Example 27 was obtained. HRMS calculated for C45H43CIFN7O6S: 863.2668, found: 432.6414 (M+2H)

Example 28: /V-|(5S_a)-5-{3-chloro-4-|2-(dimethylamino)ethoxy]-2-methylphenyl}-6-(5fluorofuran-2-yl)thieno|2,3-if]pyrimidin-4-ylj-2-hydroxy-£>-phenylalanme

Step A: 2-[2-ehloro-4-[4-chloro-6-(5-flitoro-2-furyl)thieno[2,3-à]pyrimidin-5-yl]-3methyl-phenoxy]-R, N -dimethyl-ethanamine

Using General Procedure Illb and Préparation 4e as the appropriate 6-iodo-thieno[2,3-iZ] pyrimidine dérivative and 2-(5-fluoro-2-furyl)-4,4,5,5-tetramethyi-l,3,2-dioxaborolane as the appropriate boronic acid dérivative, 2-[2-chloro-4-[4-chloro-6-(5-fluoro-2-furyl) thieno[2,3-i/]pyrimidin-5-yi]-3-methyl-phenoxy]-AÇV-dimethyl-ethanamine was obtained.

Slep B: Example 28

Using General Procedure 1b and 2-[2-chloro-4-[4-chloro-6-(5-fluoro-2-furyl)thieno[2,3-(7] pyrimidin-5-y]]-3-methyl-phenoxy]-A’,A^Ldimethyl-ethanamine as the appropriate 4-chlorothieno[2,3-i/]pyrimidine dérivative and 2-hydroxy-D-phenylalaninc as the appropriate amino acid dérivative, Example 28 was obtained as the later eluting diastereoisomer. HRMS calculated for C30H28CIFN4O5S: 610.1453, found: 611.1503 (M+H)

Example 29: 7V-|(5S_u)-5'{3-chloro-2-methyl-4-|2-(4-mcthylpiperazin-l-yl)ethoxy| phenyl}-6-(thiophcn-3-yl)thieno[2,3“i/|pyrïmidin-4-yl]-3-pyridin-2-yl-Z>-alanine and

Example 30: A-|(5/î₀)-5-{3-cliloro-2-niethyl-4-[2-(4-methylpiperazin-I-yl)ethoxy] phenyI}-6-(thiophen-3-yl)thieno[2,3-i/]pyrimidin-4-ylj-3-pyrîdin-2-yl-Z)-alanine

ORIGINAL

- 86Using General Procedure le and Préparation 4g as the appropriate 4-chloro-thieno[2,3-<Z] pyrimidine dérivative and (2Æ)-2-amino-3-(2-pyridyl)propanoic acid as the appropriate amino acid dérivative, a mixture of diastereoisomers was obtained. They were separated via preparative reversed phase chromatography using 40 mM aqueous NH₄OAc solution (pH = 4, adjusted with AcOH) and acetonitrile as eluents. Examplc 29 was obtained as the earîier eluting diastereoisomer. HRMS calculated for C32H33CIN6O3S2: 648.1744, found: 649.1811 (M+H)

Example 30 was obtained as the later eluting diastereoisomer. HRMS calculated for C3₂H₃3CIN₆O3S₂: 648.1744, found: 649.1816 (M+H)

Examplc 31: jV-[(57?₀)-5-{3-chloro-2-niethyI-4-|2-(4-methylpiperazÎn-l-yl)ethoxy| phenyl}-6-(thiophen-3-yl)thieno[2,3-i/|pyrimidin-4-yl|-3-cyclohexyl-Z)-alanine and

Example 32: /V-|(55_i,)-5-{3-chloro-2-mctliy]-4-[2-(4-methylpiperazin-l-yl)ethoxy| phcnyl}-6-(thiophen-3-yl)thieno[2,3-i/]pyrimidin-4-yl]-3-cyclohexyI-Z)-alanine

Using General Procedure le and Préparation 4g as the appropriate 4-chloro-thieno[2,3-i7] pyrimidine dérivative and (2A)-2-amino-3-cyclohexyI-propanoic acid as the appropriate amino acid dérivative, Example 31 was obtained as the earlier eluting diastereoisomer. HRMS calculated for C33I-L10CIN5O3S2: 653.2261, found: 327.6194 (M+2H)

Example 32 was obtained as the later eluting diastereoisomer. HRMS calculated for C3₃H4₀CIN₅O3S₂: 653.2261, found: 327.6195 (M+2H)

Example 33: A^r-[(57f_rt)-5-{3-chloro-2-niethyl-4-[2-(4-methylpiperazin-l-yl)ethoxy] phenyl}-6-(thiophen-3-yl)thieno[2,3-r/]pyrimÎ(lin-4-yl]-2-fIuoro-Z)-phenylalanine and

Example 34: /V-[(55₍;)-5-{3-chloro-2-methyI-4-[2-(4-metIiylpiperazÎn-l-yl)ethoxy| phenyl}-6-(thiophen-3-yl)thieno[2,3-rf]pyrinii<lin-4-yl]-2-fluoro-Z)-phenylalanine

Using General Procedure le and Préparation 4g as the appropriate 4-chloro-thieno[2,3-d] pyrimidine dérivative and (2Æ)-2-amino-3-(2-fluorophenyl)propanoic acid as the appropriate amino acid dérivative, Example 33 was obtained as the later eluting

ORIGINAL

-87diastereoisomer. HRMS calculated for C33H33CIFN5O3S2: 665.1697, found: 666.1776 (M+H)

Example 34 was obtained as the earlier eluting diastereoisomer. HRMS calculated for C33H33CIFN5O3S2: 665.1697, found: 666.1776 (M+H)

Example 35: jV-[(5/?_fl)-5-{3-chloro-2-methyl-4-[2-(morpholin-4-yl)ethoxy]phenyl}-6(thiophen-3-yl)thieno[2,3-i/|pyrimidin-4-yl|-3-pyridin-2-yl-Z)-alanine and

Example 36: 7V-[(5S_(J)-5-{3-chIoro-2-methyl-4-|2-(morpholin-4-yl)ethoxy]phenyl}-6' (thiophen-3-yl)thieno[2,3-i/]pyrimidin-4-yl]-3-pyridin-2-y!-Z>-alanine

Using General Procedure le and Préparation 4h as the appropriate 4-chloro-thieno[2,3-i7] pyrimidine dérivative and (27î)-2-amino-3-(2-pyridyl)propanoic acid as the appropriate amino acid dérivative, Example 35 was obtained as the earlier eluting diastereoisomer. HRMS calculated for C31H30CIN5O4S2: 635.1428, found: 636.1499 (M+H)

Example 36 was obtained as the later eluting diastereoisomer. HRMS calculated for C31H30CIN5O4S2: 635.1428, found: 636.1508 (M+H)

Example 37: 2-(aminomethyl)-/V-[(5Ji_tf)-5-{3-chloro-2-niethyl-4-[2-(morpholin-4-yl) ethoxy|phenyl}-6-(thiophen-3-yl)thieno|2,3-i7]pyrimidin-4-yl]-Z)-plienylalanine and

Example 38: 2-(aminomethyl)-jV-[(5S^,„)-5-{3-chloro-2-methyl-4-[2-(morpholin-4-yl) cthoxy]phenyl}-6-(thioplien-3-yI)thieno|2,3-rf] pyrimidin-4-yl]-Z>-phenylalanine

Using General Procedure le and Préparation 411 as the appropriate 4-chloro-thieno[2,3-i/| pyrimidine dérivative and (2/?)-2-amino-3-[2-(aminomethyl)phenyl]propanoic acid as the appropriate amino acid dérivative, Example 37 was obtained as the earlier eluting diastereoisomer. HRMS calculated for C33H34CIN5O4S2: 663.1741, found: 664.1808 (M+H)

Example 38 was obtained as the later eluting diastereoisomer. HRMS calculated for C33H34CIN5O4S2: 663.1741, found: 664.1825 (M+H)

ORIGINAL

-88Example 39: 7V-[(55’_£,)-5-{3-chloro-2-niethyl-4-[2-(4-methylpiperazin-l-yl)ethoxy] phenyl}-6-(prop-l-yn-l-yl)thieno[2,3-rf]pyrimidin-4-yl]-2-methoxy-Z)-phenylaIanine

Using General Procedure VI and Préparation 7b as the appropriate phénol dérivative and methanol as the appropriate alcohol dérivative, then hydrolyzing the formed intermediate according to General Procedure Vil, Example 39 was obtained. HRMS calculated for C33H36CIN5O4S: 633.2176, found: 317.6163 (M+2H)

Example 40: 2-|(l-/i?r/-buty4-lH-pyrazol-5-yl)methoxy|-A-[(5S_a)-5-{3-chloro-2methyi-4-|2-(4-methyIpiperazin-l-yI)ethoxy]phenyl}-6-(prop-l-yn-l-yl)thienoI2,3-rf] pyrimidin-4-yl]-Z)-phenylalanine

Using General Procedure VI and Préparation 7b as the appropriate phénol dérivative and Préparation C4 as the appropriate alcohol dérivative, then hydrolyzing the formed intermediate according to General Procedure Vil, Example 40 was obtained. HRMS calculated for C₄₀H4₆CIN₇O4S: 755.3021 found: 378.6573 (M+2H)

Example 41 : .V-|(5A'₍,)-5-{3-chloro-2-rnethyl-4-|2-(4-mcthylpiperazin-l-Yl)<thoxy | plienyl}-6-(prop-l-yn-l-yl)thieno|2,3-i/|pyrimidin-4-yl]-2-{[2-(2-niethoxyethyl) pyrimidin-4-yl]niethoxy}-Z)-phenylalanine

Using General Procedure VI and Préparation 7b as the appropriate phénol dérivative and Préparation C5 as the appropriate alcohol dérivative, then hydrolyzing the formed intermediate according to General Procedure VII, Example 41 was obtained. HRMS calculated for C40H44CIN7O5S: 769.2813, found: 385.6476 (M+2H).

Exaniple 42: /V-I(55'_a)-5-{3-chloro-2-methyI-4-|2-(4-methylpiperazin-l-yl)ethoxy] phenyl}-6-(prop-l-yn-l-yl)thieno[2,3-i/]pyrimidin-4-yl]-2-{[l-(2,2,2-trifluoroethyl)17/-pyrazoI-5-yI| methoxy }-£>-phenylalaninc

Using General Procedure VI and Préparation 7b as the appropriate phénol dérivative and Préparation C6 as the appropriate alcohol dérivative, then hydrolyzing the formed

ORIGINAL

-89intermediate according to General Procedure VII, Example 42 was obtained. HRMS calculated for C38H39CIF3N7O4S: 781.2425, found: 391.6300 (M+2H)

Example 43: A-|(5.S’,_;)-5-{3-chloro-2-niethyl-4-|2-(4-niethylpiperazin-l-yl)ethoxy] phenyl}-6-(prop-l-yn-l-yl)thieno[2,3-i/]pyrimidin-4-yl|-2-{[2-(morpholin-4-yl) pyrimidin-4-yl]methoxy}-Z)-phenylaIanine

Using General Procedure VI and Préparation 7b as the appropriate phénol dérivative and Préparation C7 as the appropriate alcohol dérivative, then hydrolyzing the formed intermediate according to General Procedure VII. Example 43 was obtained. HRMS calculated for C41H45CIN8O5S: 796.2922, found: 399.1546 (M+2H)

Example 44: 7V-[(55^, _fl)-5-{3-chloro-2-methyl-4-|2-(4-methylpiperazin-l-yl)ethoxy] phenyI}-6-(prop-l-yn-l-yl)thieno|2,3-i/]pyrimidin-4-yl|-2-{|2-(2,2,2-trifluoroethoxy) pyrimidin-4-yl]mcthoxy}-Z)-phenyIalanine

Using General Procedure VI and Préparation 7b as the appropriate phénol dérivative and Préparation C8 as the appropriate alcohol dérivative, then hydrolyzing the formed 15 intermediate according to General Procedure VII, Example 44 was obtained. HRMS calculated for C39H39CIF3N7O5S: 809.2374, found: 405.6262 (M+2H)

Example 45: 7V-[(55_ü)-5-{3-chloro-2-methyl-4-[2-(4-methylpiperazin-l-yl)ethoxy] phenyl}-6-(prop-l-yn-l-yI)thieno|2,3-i/]pyrimidin-4-yl]-2-{|2-(2-methoxyphenyl) pyrimidin-4-yl]methoxy}-Z)-phenylaIaninc

Using General Procedure VI and Préparation 7b as the appropriate phénol dérivative and Préparation C3 as the appropriate alcohol dérivative, then hydrolyzing the formed intermediate according to General Procedure VII, Example 45 was obtained. HRMS calculated for C44H4₄CIN₇O₅S: 817.2813, found: 409.6494 (M+2H)

ORIGINAL

- 90Example 46: jV-((55_u)-5-{3-chIoro-4-[2-(dimethylamino)ethoxy]-2-methy]phenyl}-6(prop-l-yn-l-yl)thieno[2,3-d|pyrimidin-4-yl]-2-{[l-(2,2,2-trifluoroethyl)-lH-pyrazol5-yl]methoxy}-Z>-phenylalanine

Using General Procedure VI and Préparation 7c as the appropriate phénol dérivative and Préparation C6 as the appropriate alcohol dérivative, then hydrolyzing the formed intermediate according to General Procedure VII, Exaniple 46 was obtained. HRMS calculated for CsîHmC^NôOîS: 726.2003, found: 727.2092 (M+H)

Example 47: N-|(5S,_;)-5-{3-chloro-4-|2-(diniethylaniino)etiioxy]-2-rnethyl|)heiiyl}-6(prop-l-yn-l-yl)thieno|2,3-rf] pyrimidin-4-yl|-2-{[2-(morpholin-4-yl)pyrimidin-4-yl| methoxy}-Z>-phenyialanine

Using General Procedure VI and Préparation 7e as the appropriate phénol dérivative and Préparation C7 as the appropriate alcohol dérivative, then hydrolyzing the formed intermediate according to General Procedure VII, Example 47 was obtained. HRMS calculated for C3SH40CIN7O5S: 741.2500, found: 371.6331 (M+2H)

Examplc 48: 7V-[(5S'„)-5-{3-chIoro-4-[2-(dimethylamino)ethoxy|-2-methylphcnyl}-6“ (prop-1 -yn-1 -y 1) th ie no [ 2,3-zZ] py rimidin-4-y 11-2- {12-(2,2,2-trifluoroethoxy)pyrimidin-4yl)methoxy}-£>-phenylalanine

Using General Procedure VI and Préparation 7c as the appropriate phénol dérivative and Préparation C8 as the appropriate alcohol dérivative, then hydrolyzing the formed intermediate according to General Procedure VII, Example 48 was obtained. HRMS calculated for CatJ-^ClFjNôOÿS: 754.1952, found: 755.1971 (M+H)

Example 49: jV-[(55_£,)-5-{3-chIoro-4-[2-(dimcthylamino)ethoxy]-2-methyIphenyI}-6(prop-l-yn-l-yl)thieno[2,3-if]pyrimidin-4-yl|-2-{[2-(2-methoxyphenyl)pyrimidin-4-yl| methoxy}-Z>-phenylalanine

ORIGINAL

-9l

Using General Procedure VI and Préparation 7c as the appropriate phénol dérivative and Préparation C3 as the appropriate alcohol dérivative, then hydrolyzing the formed intermediate according to General Procedure VII, Exaniple 49 was obtained. HRMS calculated for C41H39CIN6O5S: 762.2391, found: 371.6323 (M+2H)

Example 50: /V-[(55'_u)-5-{3-chloro-4-[2-(dimcthylamino)ethoxy|-2-methylphcnyl}-6-(4fiuorophenyl)thienoI2,3-i/]pyrimidin-4-yl]-2-{|2-(2-fIuorophenyl)pyrimidin-4-yl| methoxy}-Z>-phenyIalanine

Step A: ethyl (2^)-2-[[5-bromo-6-(4-fluorophenyl)thieno[2,3-à]pyrimidin-4-yl]amino]-3[2-[[2-(2-fl uorophenyl)pyrimidin-4-yl] methoxy]phenyl]propanoate

Using General Procedure VI and Préparation 7d as the appropriate phénol dérivative and Préparation C9 as the appropriate alcohol dérivative, ethyl (27?)-2-[[5-bromo-6-(4fluorophenyl)thieno[2,3-i/]pyrimidin-4-yl]amino]-3-[2-[[2-(2-fluorophenyl)pyrimidin-4-yl] methoxy]phenyl]propanoate was obtained. ’H NMR (400 MHz, DMSO-dô) δ: 8.84 (d, IH), 8.39 (s, IH), 7.95 (td, IH), 7.58-7.52 (m, 3H), 7.39-7.24 (m, 8H), 7.13 (d, IH), 6.95 (t, IH), 5.29-5.15 (m, 3H), 4.16 (q, 2H), 3.63 (dd, IH), 3.25 (dd, 1 H), 1.19 (t, 3H)

Step B: ethyl (2R)-2-[[5-[3-chloro-4-(2-dimethylaminoethyloxy)-2-methyl-phenyl]-6-(4fluorophenyl)thieno[2,3-à]pyrimidin-4-yl]amino]-3-[2-[[2-(2-fluorophenyl)pyrimidin-4yl]methoxy]phenyl]propanoate eq. ethyl (27?)-2-[[5-bromo-6-(4-fluorophenyl)thieno[2,3-i7}pyrimidin-4-yl]amino]-3-[2[[2-(2-fluorophenyl)pyrimidin-4-yl]methoxy]phenyl|propanoate and 1.2 eq. Préparation B5 were dissolved in dioxan (5 mL/mmol), then 5 mol% AtaPhos, 3 eq. CS2CO3 and water(5 mL/mmol) were added and the mixture was stirred at 70 °C under argon atmosphère until no further conversion was observed. Then the mixture was diluted with EtOAc and was washed with brine. The organic phase was dried over Na₂SO4, filtered and the filtrate was concentrated under reduced pressure. The crude product was purified via flash chromatography using DCM and MeOH as eluents to obtain ethyl (2/?)-2-[[5-[3chloro-4-(2-dimethylaminoethyloxy)-2-methyl-phenyi]-6-(4-fluorophenyl)thieno[2,3-i/] pyrimidin-4-yl]amino]-3-[2-[[2-(2-fluorophenyl)pyrimidin-4-yl]methoxy]phenyl] propanoate as a mixture of diastereoisomers. MS (M+H): 834.6

ORIGINAL

-92Step C: Example 50

Using General Procedure VII and ethyl (27?)-2-[[5-[3-chloro-4-(2-dimethylaminoethyl oxy)-2-methyl-phenyl]-6-(4-fluorophenyl)thieno[2,3-i/]pyriniidin-4-yl]amino]-3-[2-[[2-(2fluorophenyl)pyrimidin-4-yl]methoxy]phenyI]propanoate as the appropriate ester dérivative, Example 50 was obtained as the later eluting diastereoisomer. HRMS calculated for C₄3H37ClF₂N₆O₄S: 806.2254, found: 807.2343 (M+H)

Example 51: 7V-[(5S_û)-5-{3-chloro-4-|2-(dimethylamino)ethoxy]-2-methylphenyl}-6-(4fluorophenyl)thieno|2,3-rf|pyrimïdin-4-ylj-2-({2-[2-(2-methoxyetlioxy)plienyl] pyrimidin-4-yl}methoxy)-Z)-phenylalanine

Step A: ethyl (2R)-2-[[5-bromo-6-(4-fluorophenyl)thieno[2,3-d]pyrimidin-4-yl]amino]-3[2-[[2-[2-(2-methoxyethoxy)phenyl]pyrimidin-4-yl]methoxy]phenyl]propanoate

Using General Procedure VI and Préparation 7d as the appropriate phénol dérivative and Préparation CIO as the appropriate alcohol dérivative, ethyl (27î)-2-[[5-bromo-6-(4fluorophenyl)thieno[2,3-i/]pyrimidin-4-yl]aniino]-3-[2-[[2-[2-(2-methoxyethoxy)phenyl] pyrimidin-4-yl]methoxy]phenyl]propanoate was obtained. *H NMR (400 MHz, DMSO-dô) δ: 8.80 (d, IH), 8.41 (s, IH), 7.57-7.53 (m, 3H). 7.46-7.23 (m, 7H), 7.16 (d, IH), 7.07 (d, IH), 7.03 (t, IH), 6.94 (t, IH), 5.28-5.23 (m, IH), 5.19 (dd, 2H), 4.18-4.11 (m, 4H), 3.61-

3.57 (m, 3H), 3.27 (dd, 1 H), 3.21 (s, 3H), 1.19 (t,3H)

Step B: ethyl (2'R.)-2-[[5-[3-chloi-o-4-(2-dimethylaminoethyloxy)-2-methyl-phenyl]-6-(4fluorophenyl)lhieno[2,3-à]pyrimidin-4-yl]amino]-3-[2-[[2-[2-(2-methoxyethoxy)phenyl] pyi ‘i midin-4 -yl]metl ioxy]phenyljpropanoate eq. ethyl (2Æ)-2-[[5-bromo-6-(4-iluorophenyI)thieno[2,3-i/]pyrimidin-4-yl]amino]-3-[2[[2-[2-(2-methoxyethoxy)phenyl]pyrimidin-4-yl]methoxy]phenyl]propanoate and 1.2 eq. Préparation B5 were dissolved in dioxan (5 mL/mmol), then 5 mol% AtaPhos, 3 eq. CS2CO3 and water(5 mL/mmol) were added and the mixture was stirred at 70°C under argon atmosphère until no further conversion was observed. Then the mixture was diluted with EtOAc and was washed with brine. The organic phase was dried over Na₂SO₄, filtered and the filtrate was concentrated under reduced pressure. The crude product was purified

ORIGINAL

-93 via flash chromatography using DCM and MeOH as eluents to obtain ethyl (27î)-2-[[5-[3-chloro-4-(2-dimethylaminoethyloxy)-2-methyl-phenyl]-6-(4-fluorophenyl) thieno[2,3-i7]pyrimidin-4-yl]amino]-3-[2-[[2-[2-(2-methoxyethoxy)phenyl]pyrimidin-4-yl] methoxy]phenyl]propanoate as a mixture of diastereoisomers. MS (M+H): 890.6

Step C: Example 51

Using General Procedure VU and ethyl (27î)-2-[[5-[3-chloro-4-(2-dimethylaminoethyi oxy)-2-methyl-phenyl]-6-(4-fluorophenyl)thieno[2,3-i/]pyrimidin-4-yl]amino]-3-[2-[[2-[2(2-methoxyethoxy)phenyl]pyrimidin-4-yl]methoxy]phenyl]propanoate as the appropriate ester dérivative, Example 51 was obtained as the iater eluting dîastereoisomer. HR.MS calculated for C₄6H44C1FN₆O₆S: 862.2716, found: 432.1442 (M+2H)

Example 52: A^r-|(57?_û)-5-(3,5-dichIoro-4-hydroxy-2-metliylphenyl)-6-ethylthicno[2,3-i/| pyrimidin-4-yi]-Z)-phenylalanine and

Example 53: A'-|(55₁₇)-5-(3,5-dichloro-4-hydiOXV-2-nicthylphenyl)-6-ethylthieno|2,3-i7| pyrimidin-4-yl]-D-phenylalanine

Using General Procedure Vil and Préparation 7f as the appropriate ester dérivative, a mixture of diastereoisomers was obtained. They were separated via préparative reversed phase chromatography using 0.1 % aqueous TFA solution and acetonitrile as eluents. Example 52 was obtained as the earlier eluting diastereomer. HR.MS calculated for C24H21CI2N3O3S: 501.0681, found: 502.0755 (M+H)

Examplc 53 was obtained as the later eluting diastereomer. HRMS calculated for C24H21CI2N3O3S: 501.0681, found: 502.0772 (M+H)

Example 54: .5-((55,,)-5-(3,5-dichloro-2-metliyl-4-|2-(4-methylpiperazin-l-yl)ethoxy] phenyl}-6-ethylthieno|2,3-rf]pyrimidin-4-yl)-Z)-phenylalanine and

Example 55: A-((5/?,,)-5-{3,5-dichloro-2-methyl-4-|2-(4-metliylpipeiazin-1 -yl)ethoxy] phenyl}-6-ethylthieno[2,3-i/|pyrimidin-4-yl)-Z)-plienylalanine

ORIGINAL

-94Using General Procedure VI and Préparation 7f as the appropriate phénol dérivative and 2-(4-methylpiperazin-l-yl)ethanol as the appropriate alcohol dérivative, then hydrolyzing the formed intermediate according to General Procedure VII, Exaniple 54 was obtained as the later eluting diastereoisomer. HRMS calculated for C31H35CI2N5O3S: 627.1838, found: 5 628.1935 (M+H)

Example 55 was obtained as the earlier eluting diastereoisomer. HRMS calculated for C31H35CI2N5O3S: 627.1838, found: 628.1932 (M+H)

Example 56: jV-{(5S_o)-5-[3-chloro-4-(2-hydroxycthoxy)-2-methylphenyl|-6-ethyl thieno[2,3-rf]pyrimidin-4-yl}-Z>-phenylalanine iO Using General Procedure VI and Préparation 7gdl as the appropriate phénol dérivative and 10 eq. ethylene glycol as the appropriate alcohol dérivative, then hydrolyzing the formed intermediate according to General Procedure VII, Example 56 was obtained. HRMS calculated for C₂₆H₂₆CIN₃O4S: 51 1.1333, found: 512.1390 (M+H)

Example 57: /V-{(55„)-5-[4-(carboxymethoxy)-3-chloro-2-methylphenyI]-6-ethylthieno 15 [2,3-i/|pyrimidin-4-yl)-Z)-phenylalaninc

Using General Procedure VI and Préparation 7gdl as the appropriate phénol dérivative and 2-hydroxy-/V,/V-dimethylacetamide as the appropriate alcohol dérivative, then hydrolyzing the formed intermediate according to General Procedure Vil, Example 57 was obtained. HRMS calculated for C₂6H₂₄C1N₃O₅S: 525.1125, found: 526.1217 (M+H)

Example 58: 7V-((5/f„)-5-{3-cliIoro-4-[2-(dimetliylamino)ethoxy]-2-metliylphenyl}-6ethylthieno[2,3-i/|pyrimidin-4-yl)-L-plienyIalanine and

Exaniple 59: 2V-((55_n)-5-{3-chloro-4-[2-(dimethylamino)ethoxy]-2-metliylphenyl}-6etliylthieno[2,3-rfj pyrimidin-4-yI)-L-phenylalanine

Using General Procedure VI and Préparation 7g as the appropriate phénol dérivative and 2-(dimethylamino)ethanol as the appropriate alcohol dérivative, then hydrolyzing the

ORIGINAL

-95formed intermediate according to Généra! Procedure VII, a mixture of diastereoisomers was obtained. They were separated via préparative reversed phase chromatography using 40 mM aqueous NH4OAC solution (pH = 4, adjusted with AcOH) and acetonitrile as eluents. Example 58 was obtained as the earlier eluting diastereoisomer. HRMS calculated for C28H31CIN4O3S: 538.1805, found: 539.1869 (M+H)

Example 59 was obtained as the later eluting diastereoisomer. HRMS calculated for C28H31CIN4O3S: 538.1805, found: 539.1866 (M+H)

Example 60: A^r-((55„)-5-{3-chIoro-4-(3-(dimethylamino)propoxy]-2-methylphenyl}-6ethylthieno|2,3-i/|pyrimidin-4-yl)-Z)-phenyIaIanine

Using General Procedure VI and Préparation 7gdl as the appropriate phénol dérivative and 3-(dimethylamino)propanol as the appropriate alcohol dérivative, then hydrolyzing the formed intermediate according to General Procedure VII, Example 60 was obtained. HRMS calculated for C^HjaCl^C^S: 552.1962, found: 553.2036 (M+H)

Example 61: A^r-((55’_fl)-5-{3-chloro-2-methyl-4-[2-(niorpholin-4-yl)ethoxy|plienyI}-615 ctiiylthieno|2,3-i/|pyrimidin-4-yl)-Z>-phenylalanine

Using General Procedure VI and Préparation 7gdl as the appropriate phénol dérivative and 2-morpholinoethanol as the appropriate alcohol dérivative, then hydrolyzing the formed intermediate according to General Procedure VII, Exampie 61 was obtained. HRMS calculated for C30H33CIN4O4S: 580.1911, found: 581.1981 (M+H)

Exampie 62: /V-((55_a)-5-{3-chloro-2-methyl-4-[2-(4“methylpiperazin-l-yl)ethoxy] phenyI}-6-ethylthieno[2,3-rf]pyrimidin-4-yl)-Z)-phenylalanine and

Example 63 7V-((5/î_a)-5-{3-chloro-2-methyl-4-[2-(4-methylpiperazin-l-yl)cthoxy| phenyI}-6-ethyIthieno[2,3-rf]pyrimidin-4-yl)-£)-phenylalanine

Using General Procedure VI and Préparation 7g as the appropriate phénol dérivative and 2-(4-methylpiperazin-l-yl)ethanoi as the appropriate alcohol dérivative, then hydrolyzing

ORIGINAL

-96the formed intermediate according to General Procedure Vil, Example 62 was obtained as the earlier eluting diastereoisomer. HRMS calculated for C3JH36CIN5O3S: 593.2227, found: 594.2313 (M+H)

Example 63 was obtained as the later eluting diastereoisomer. HRMS calculated for 5 C3₁H₃₆C1N₅C)3S: 593.2227, found: 594.2304 (M+H)

Example 64: 2V-((55_e)-5-{3-chloro-2-methyl-4-[3-(4-methylpiperazin-I-yl)propoxy| phenyl}-6-ethylthieno[2,3-rf[pyrimidin-4-yl)-/)-phenylalanine

Using General Procedure VI and Préparation 7gdl as the appropriate phénol derivative and 3-(4-methylpiperazin-l-yl)propan-l-ol as the appropriate alcohol derivative, then 10 hydrolyzing the formed intermediate according to General Procedure Vil, Example 64 was obtained. HRMS calculated for C32H38CIN5O3S: 607.2384, found: 608.2444 (M+H)

Example 65: ,V-((5A'_Î;)-5-|3-chloro-2-niethyl-4-|2-(4-inethylpiperazin-1 -yl)ethoxy | phenyl}-6-[5-(methoxycarbonyl)-4-niethylfuran-2-yl|thieno[2,3-rf]pyrimidÎn-4-yl)-2methoxy-D-phcnylalaninc and

Example 66: A^r-((5Æ„)-5-{3-chloro-2-methyl-4-[2-(4-methylpiperazin-l-yl)ethoxy] phenyl}-6-|5-(metlioxycarbonyl)-4-niethylfuran-2-yI]tliieno[2,3-rZ]pyrimidin-4-yl)-2methoxy-D-phenylalanine

Step A: [4-chloro-5-[3-chloro-2~methyl-4-[2-(4-niethylpiperazin-l-yl)ethoxy]phenyl] 20 thieno[2,3-<3]pyrimidin-6-yl]-triinethyl-stannane

1.97 g 4-ch]oro-5-[3-chloro-2-methyl-4-[2-(4-methylpiperazin-1 -yl)ethoxy]phenyl]thieno [2,3-i7]pyrimidine (4.50 mmol, from Step A of Préparation 4a) was dissolved in 40 mL dry THF under N2 atmosphère, and the mixture was cooled to -78 °C. Then 4.5 mL LDA (9 mmol, 2M solution in heptane, THF and ethyl benzene) was added and the mixture was 25 stirred at -78 °C for 1 hour. Then 13.5 mL Me3SnCl solution (13.5 mmol, IM in hexane) was added and the mixture was allowed to warm up to r.t. The mixture was then diluted with cc. NH4CI solution and extracted with diethyl ether. The organic phase was dried over Na2SO4, fîitered and the filtrate was concentrated under reduced pressure. Then it was

ORIGINAL

-97dissolved in 60 mL EtOAc, and 40 mL saturated NaF solution was added, and it was stirred at r.t. ovemight. Then it was filtered, the phases of the filtrats were separated. The aqueous phase was extracted with EtOAc. The combined organic phases were dried over NaiSO-i, filtered and the filtrate was concentrated under reduced pressure. The crude product was purified via flash chromatography using DCM and MeOH as eluents to obtain [4-chloro-5-[3-chloro-2-methyl-4-[2-(4-methylpiperazin-l-yl)ethoxy] phenyl]thieno[2,3-<7] pyrimidin-6-yl]-trimethyl-stannane. HRMS calculated for CijHjoN^OSCLSn: 600.0539, found: 601.0584 (M+H)

Sien B: methyl 5-[4-chloro-5-[3-chloro-2-methyl-4-[2-(4-meihylpiperazin-l-yl)ethoxy] phenyl]thieno[2,3-à]pyiiniidin-6-yl]-3-methyl-furan-2-caihoxylate

900 mg [4-chloro-5-[3-chloro-2-methyl-4-[2-(4-methylpiperazin-1 -yl)ethoxy]phenyl] thieno[2,3-i7]pyrirnidïn-6-yl]-trirnethyl-stannane (1.50 mmol), 657 mg methyl 5-bromo-3methyl-furan-2-carboxylate (3 mmol), 29 mg Cul (0.15 mmol), 29 mg Pd(PhCN)2Cl₂(0.075 mmol), 46 mg Ph₃As (0.15 mmol) and 2 mL NMP were stirred at 100 °C until no further conversion was observed. Then the mixture was diluted with EtOAc and washed with saturated NaF solution. The aqueous phase was extracted with EtOAc. The combined organic phases were dried over Na2SO.|, filtered and the filtrate was concentrated under reduced pressure. The crude product was purified via flash chromatography using DCM and MeOH as eluents to obtain methyl 5-[4-chloro-5-[3-chIoro-2-methyl-4[2-(4-methylpiperazin-l-yl)ethoxy]phenyl]thieno[2,3-i/|pyrimidin-6-yl]-3-methyl-furan-2carboxylate. HRMS calculated for C27H28CI2N4O4S: 574.1208, found: 575.1263 (M+H)

Step C: Example 65

Using General Procedure lb and methyl 5-[4-chloro-5-[3-chloro-2-methyL4-[2-(4-methyl piperazin-l-yl)ethoxy]phenyl]thieno[2,3-t/]pyrimidin-6-yl]-3-methyl-furan-2-carboxylate as the appropriate 4-chloro-thieno[2,3-<7]pyrimidine dérivative and (2Jf)-2-amino-3-(2methoxyphenyl)propanoic acid as the appropriate amino acid dérivative Example 65 was obtained as the earlier eluting diastereoisomer. HRMS calculated for C37H40CIN5O7S: 733.23369, found: 367.6263 (M+2H)

Example 66 was obtained as the later eluting diastereoisomer. F1RMS calculated for C37H40CIN5O7S: 733.23369, found: 367.6223 (M+2H)

ORIGINAL

-98Example 67: /V-lô-ethyl-S-ÎS-hydroxy-l-methylphenylJthieno^^-rfJpyrimidin^-yll-Dphenylalanine

Using General Procedure Hd and Préparation 3a as the appropriate 5-iodo-thieno[2,3-t/| pyrimidine dérivative and 2-methyl-3-(4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2-yI)phenoI as the appropriate boronic acid dérivative. Example 67 was obtained as a mixture of diastereoisomers. HRMS calculated for C24H23N3O3S: 433.1460, found: 434.1545 and 434.1535 (M+H)

Example 68: /V-|6-ethyl-5-(3-lluoro-2-niethylphenyl)thieno[2,3-rf|pyrimîdin-4-yl|-Z)phenylalanine

Using General Procedure Ha and Préparation 3a as the appropriate 5-iodo-thieno[2,3-i/] pyrimidine dérivative and (3-fluoro-2-methyl-phenyl)boronic acid as the appropriate boronic acid dérivative, Example 68 was obtained as a mixture of diasteromeres. HRMS calculated for C74H22FN3O2S: 435.1417, found: 436.1489 and 436.1484 (M+H)

Example 69: 7V-|6-ethyl-(5S'_(J)-5'-(3-fluoro-2-niethylphenyI)thieno[2,3-i/]pyrimidin-4yl]-Z>-phenyIalanine and

Example 70: /V-|6-ethyI-(5Æ„)-5-(3-fluoro-2-methylphenyl)thieno|2,3-</|pyrimidin-4yl]-Z>-phenylalanine

Diastereoisomers of Example 68 were separated by preparative reversed phase chromatography using 25 mM aqueous NH4HCO3 solution and acetonitrile as eluents. Example 69 was obtained as the earlier eluting diastereoisomer. HRMS calculated for C24H22FN3O2S: 435.1417, found: 436.148] (M+H)

Example 70 was obtained as the later eluting diastereoisomer. HRMS calculated for C24H22FN3O2S: 435.1417, found: 436.1498 (M+H)

ORIGINAL

-99Example 71: /V-[6-ethyl-5-(lH-indoI-7-yl)thieno[2,3-i/]pyrimidin-4-yl]-Z>phenylalanine, diastereoisomer 1 and

Example 72:7V-[6-ethyI-5-(177-indol-7-yI)thieno[2,3-rf]pyrimidin-4-yl|-/>phenylalanine, diastereoisomer 2

Using General Procedure lia and Préparation 3a as the appropriate 5-iodo-thieno[2,3-<Y] pyrimidine dérivative and 7-(4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2-yl)-l/7-indole as the appropriate boronic acid dérivative, a mixture of diastereoisomers was obtained. They were separated via préparative reversed phase chromatography using 0.1 % aqueous TFA solution and acetonitrile as eluents. Example 71 was obtained as the earlier eluting diastereoisomer. HRMS calculated for C25H22N4O2S: 442.1463, found: 443.1540 (M+H) Example 72 was obtained as the later eluting diastereoisomer. HRMS calculated for C25H22N4O2S: 442.1463, found: 443.1537 (M+H)

Example 73: jV-[6-ethyl-(55_a)-5-(l/7-indoI-4-yI)thieno[2,3-rf|pyrimidin-4-yl]-Z)phenylalanine and

Example 74: A-[6“Cthyl-(5Æ_a)-5-(lH-indol-4-yl)thieno[2,3-i/|pyrimidin-4-vI]-Z)phenylalanine

The diastereoisomers of Préparation 7h were separated by préparative reversed phase chromatography using 0.1 % aqueous TFA solution and acetonitrile as eluents. Example 73 was obtained as the earlier eluting diastereoisomer. HRMS calculated for C25H22N4O2S: 442.1463, found: found: 443.1529 (M+H)

Example 74 was obtained as the later eluting diastereoisomer. HRMS calculated for C₂5H22N₄O₂S: 442.1463, found: 443.1538 (M+H)

Example 75: j'V-|6-ethyl-(55_ff)-5-(3-methoxy-2-methylphenyI)thienol2,3-i/]pyrimidin-4yl]-D-phenylalanine and

ORIGINAL

- ιοοExample 76: A'-[6-ethyl-(5/?j)-5-(3-inethoxy-2-niethylphenyl)thieno|2,3-i/]pyrimidin4-yI]-£>-phenylalanine

Using General Procedure Ilb and Préparation 3a as the appropriate 5-iodo-thieno[2,3-iZ] pyrimidine dérivative and 2-(3-methoxy-2-methyl-phenyl)-4,4,5,5-tetramethyl-l,3,2dioxaborolane as the appropriate boronic acid dérivative, using DME:water 5:1 instead of 2-Me-THF, and separating the diastereoisomers by préparative reversed phase chromatography using 25 mM aqueous NH4HCO3 solution and acetonitrile as eluents, Example 75 was obtained as the earlier eluting diastereoisomer, HRMS calculated for C25H25N3O3S: 447.1617, found: 448.1701 (M+H)

Example 76 was obtained as the later eluting diastereoisomer. HRMS calculated for C25H25N3O3S: 447.1617, found: 448.1672 (M+H)

Example 77:7V-|(5/ï_a)-5-(2-chloro-3-methylpyridin-4-yI)-6-ethyItliieno[2,3-rf] pyrimidin-4-yl]-Z)-phenylalanine and

Example 78 A^r-|(55j-5-(2-chloro-3-niethylpyridin-4-vl)-6-ethylthieno|2,3-<7] pyrimidin-4-yl]-Z)-phenylalanine

Using General Procedure Ilb and Préparation 3a as the appropriate 5-iodo-thieno[2,3-i7] pyrimidine dérivative and 2-chloro-3-methyl-4-(4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2yDpyridine as the appropriate boronic acid dérivative, using DME:water 5:1 instead of 2-Me-THF, and separating the diastereoisomers by préparative reversed phase chromatography using 0.1 % aqueous TFA solution and acetonitrile as eluents, Example 77 was obtained as the earlier eluting diastereoisomer. HRMS calculated for C23H21CIN4O2S: 452.1074, found: 453.1158 (M+H)

Exampie 78 was obtained as the later eluting diastereoisomer. HRMS calculated for C23H21CIN4O2S: 452.1074, found: 453.1165 (M+H)

Example 79: A^r-[6-ethyl-5-(naphthaIen-l-yl)thieno[2,3-i/]pyrimidin-4-yI|-£>phenylalanine

ORIGINAL

- ΙΟΙ Using General Procedure Ilb and Préparation 3a as the appropriate 5-iodo-thieno[2,3-i/] pyrimidîne dérivative and 4,4,5,5-tetramethyl-2-(l-naphthyl)-l,3.2-dioxaborolane as the appropriate boronic acid dérivative, a mixture of diastereoisomers was obtained. It was purified via preparative reversed phase chromatography using 0.1 % aqueous TFA solution 5 and acetonitrile as eluents, Exaniple 79 was obtained as the mixture of diastereoisomers.

HRMS calculated for C₂₇H₂3N₃O₂S: 453.151 1, found: 454.1580 and 454.1580 (M+H)

Example 80: jV-|6-ethyl-5-(quinolin-5-yl)thieno|2,3-rf]pyrimidin-4-yl|-/)phenylalanine

Using General Procedure Ha and Préparation 3a as the appropriate 5-iodo-thieno[2,3-</J 10 pyrimidîne dérivative and 5-(4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2-yl)quinolone as the appropriate boronic acid dérivative, a mixture of diastereoisomers was obtained. It was purified via preparative reversed phase chromatography using 40 mM aqueous NH4OAC solution (pH = 4, adjusted with AcOH) and acetonitrile as eluents. Example 80 was obtained as amixture of diastereoisomers. HRMS calculated for C₂6H₂2N₄O2S: 454.1463, 15 found: 455.1554 and 455.1518 (M+H)

Example 81: A^r-[6-ethyl-(5.S'_ff)-5-(isoquinoIin-4-yI)thieno|2,3-if|pyrimidin-4-ylj“Z)“ phenylalanine and

Example 82: 7V-(6-ethyL(5/f_a)-5-(isoquinolin-4-yl)thieno[2,3-rf]pyriniidin-4-yl]-Z>20 phenylalanine

Step A: 4-chloro-6-ethyl-5~(4-isoquinolyl)thieno[2.3-à]pyrimidîne

Using General Procedure Ile and Préparation 2a as the appropriate 5-iodo-thieno[2,3-c/] pyrimidin and 4-(4,4,5,5-tetramethyl-l ,3,2-dioxaborolan-2-yl)isoquinoline as the appropriate boronic acid dérivative, 4-chloro-6-ethyl-5-(4-isoquinolyl)thieno[2,3-c/] 25 pyrimidîne was obtained.

’H NMR (500 MHz, DMSO-d₆) δ: 9.46 (s, IH), 8.93 (s, IH), 8.50 (s, 1 H), 8.26 (m, IH), 7.74 (m, 2H), 7.42 (m, IH), 2.65 (q, 2H), 1.14 (t, 3H)

HRMS calculated for C17H12CIN3S: 325.0440; found 326.0502 (M+H)

ORIGINAL

- 102Step B: N-[6-ethyl-5-(iso(pünolin-4~yl)thieno[2,3-d]pyrimidin-4-yl]-D-phenylalanine

Using General Procedure la, the product of Step A as the appropriate 4-chloro-thieno [2,3-d]pyrimidine dérivative and £>-phenylalanine as the appropriate amino acid dérivative A-[6-ethyl-5-(isoquinolin-4-yl)thieno[2,3-i/]pyrimidin-4-yl]-£)-phenylalanine was obtained as a mixture of diastereoisomers. They were separated via preparative reversed phase chromatography using water and acetonitrile as eluents. Example 81 was obtained as the earlier eluting diastereoisomer. HRMS calculated for C26H22N4O2S: 454.1463, found: 455.1526 (M+H)

Example 82 was obtained as the later eluting diastereoisomer. HRMS calculated for C26H22N4O2S: 454.1463, found: 455.1538 (M+H)

Example 83: /V-|6-ethyl-(5S_α)-5-(l-mcthyl-177-indol·7-yl)thieno[2,3-ί7]pyrimidin-4-yl]£>-phenylalanine and

Example 84: /V-|6-ethyl-(5Æ_u)-5-(l-methyl-l/7-indol-7-yl)thieno[2,3-i/]pyrimidin-4-yl|D-phenylalanine

Using General Procedure 11b and Préparation 3a as the appropriate 5-iodo-thieno[2,3-i/] pyrimidine dérivative and l-methyl-7-(4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2-yl)indole as the appropriate boronic acid dérivative, a mixture of diastereoisomers was obtained. They were separated via preparative reversed phase chromatography using 40 mM aqueous NH4OAC solution (pH = 4, adjusted with AcOH) and acetonitrile as eluents. Example 83 was obtained as the earlier eluting diastereoisomer. HRMS calculated for C26H24N4O7S: 456.1620, found: 457.1671 (M+H)

Example 84 was obtained as the later eluting diastereoisomer. HRMS calculated for C26H22N4O2S: 456.1620, found: 457.1701 (M+H)

Example 85: 7V-|6-ethyl-(5S_B)-5-(3-methyl-l/7-indol-4-yl)thieno[2,3-r/]pyrimidÎn-4-yl|Z)-plienylalanine and

ORIGINAL

- 103 Example 86: /V-[6-ethyl-(5Æ_iZ)-5-(3-methyl-l //-indoI-4-yl)thicno|2.3-i7]pyrimidin-4-yl|D-phenylalanine

Using General Procedure Ile and Préparation 3a as the appropriate 5-iodo-thieno[2,3-i7] pyrimidine dérivative and 3-methyl-4-(4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2-yl)-lH5 indole as the appropriate boronic acid dérivative, a mixture of diastereoisomers was obtained. They were separated via préparative reversed phase chromatography using 40 mM aqueous NH₄OAc solution (pH = 4, adjusted with AcOH) and acetonitrile as eluents. HRMS calculated for C26H24N4O2S: 456.1620, found: 457.1691 (M+H)

Example 86 was obtained as the later eluting diastereoisomer. HRMS calculated for 10 C26H24N4O2S: 456.1620, found: 457.1688 (M+H)

Example 87: N-[6-cthyl-5-( 1 -methyl-lH-indazol-4-yl)thieno[2,3-</] pyrimidin-4-yl|-Z>phenylalanine

Using General Procedure Ha and Préparation 3a as the appropriate 5-iodo-thieno[2,3-t/] pyrimidine dérivative and l-methyl-4-(4,4,5,5-tetramethyl-l,3,2-dioxaborolan-215 yl)indazole as the appropriate boronic acid dérivative, Example 87 was obtained as a mixture of diastereoisomers. HRMS calculated for C25H23N5O2S: 457.1572, found: 458.1646 and 458.1648 (M+H)

Example 88: /V-|6-ethyl-(55_a)-5-(l-methyl-l//-indazol-7-yl)thicno|2,3-i/]pyrimidin-4yl]-Z>-phenylalanine and

Example 89: 7V-[6-ethyl-(57î_a)-5-(l-methyl-lH-indazol-7-yl)thieno|2,3-J]pyrimidm-4yl]-Z)-plienyIaIanine

Using General Procedure lia and Préparation 3a as the appropriate 5-iodo-thieno[2,3-i/] pyrimidine dérivative and (1-methylindazol-7-yl)boronic acid as the appropriate boronic 25 acid dérivative, Example 88 was obtained as the earlier eluting diastereoisomer. HRMS calculated for C25H23N2O2S: 457.1572, found: 458.1641 (M+H)

ORIGINAL

- I04Example 89 was obtained as the later eluting diastereoisomer, HRMS calculated for C25H₂3N₂O₂S: 457.1572, found: 458.1634 (M+H)

Examplc 90: .'V-|(55_i,)-5-(3-chloro-4-hvdroxy-2-meth)lphenyl)-6-ethylthieno|2,3-i/j pyrimidin-4-yl]-Z)-phenyIalanine and

Example 91: A-[(57f_a)-5-(3-chloro-4-hydroxy-2-niethylphenyl)-6-ethylthieno[2,3-rf] pyrimidin-4-yl]-Z)-phenylaIanine

500 mg Préparation 7e (1.12 mmol) and 157 mg NCS (1.173 mmol) were dissolved in mL THF and the mixture was stirred at 60 °C overnight. The solvent was evaporated 10 under reduced pressure and the residue was purified via flash chromatography using heptane and EtOAc as eluents to obtain methyl 2-[[5-(3-chloro-4-hydroxy-2-methylphenyl)-6-ethyl-thieno[2,3-i/J pyrimidin-4-yl]amino]-3-pheny]-propanoate as a mixture of diastereoisomers (along with other regioisomers). The crude mixture was hydrolysed according to General Procedure VII. The diastereoisomers were purified and separated via 15 préparative reversed phase chromatography using 25 mM aqueous NH4HCO3 solution and acetonitrile as eluents. Example 90 was obtained as the later eluting diastereoisomer. HRMS calculated for C24H₂₂C1N3O3S: 467.1070, found: 468.1153 (M+H)

Exaniple 91 was obtained as the earlier eluting diastereoisomer. HRMS calculated for C24H22CIN3O3S: 467.1070, found: 468.1143 (M+H)

Example 92: ;V-|(57?,,)-5-(2,3-dicldoiophenvl)-6-ethvlthieno|2,3-<7|pyrimidin-4-yl]-Z)phenylalanine and

Examplc 93: j^,V-[(5S„)-5-(2,3-dichlorophenyl)-6-ethylthieno[2,3-i/]pyriniÎdin-4-yl]-Z)phenylalanine

Using General Procedure lia and Préparation 3a as the appropriate 5-iodo-thieno[2,3-<7] pyrimidine derivalive and (2,3-dichlorophenyl)boronic acid as the appropriate boronic acid dérivative, using Xantphos instead of BuPAd₂, a mixture of diastereoisomers was obtained. They were separated via préparative reversed phase chromatography using 0.1 %

ORIGINAL

-105aqueous TFA solution and acetonitrile as eluents, Example 92 was obtained as the earlier eiuting diastereoisomer. HRMS calculated for C23H19CI2N3O2S: 471.0575, found: 472.0667 (M+H)

Example 93 was obtained as the later eiuting diastereoisomer. HRMS calculated for C23H19CI2N3O2S: 471.0575, found: 472.0654 (M+H)

Example 94: jV-[(57î_fl)-5-(3,4-dichloro-2-niethylphenyl)-6-ethylthieno|2,3-i/|pyrimidin4-yl]-D-phenylalanine and

Example 95: /V-[(55_fl)-5-(3,4~dichloro-2-mcthylphenyl)-6-ethyIthieno[2,3-rf]pyrimidÎn4-yl]-Z>-phenylalanine

Using General Procedure Ilb and Préparation 3a as the appropriate 5-iodo-thieno[2,3-cZj pyrimidine dérivative and (3,4-dichloro-2-methyl-phenyl)boronic acid as the appropriate boronic acid dérivative, using Xantphos as ligand instead of Q-Phos and DME:water 4:1 instead of 2-Me-THF, a mixture of diastereoisomers was obtained. They were separated via preparative reversed phase chromatography using 0.1 % aqueous TFA solution and acetonitrile as eluents, Example 94 was obtained as the earlier eiuting diastereoisomer. HRMS calculated for C24H21CI2N3O2S: 485.0731, found: 486.0816 (M+H)

Example 95 was obtained as the later eiuting diastereoisomer. HRMS calculated for C24H21CI2N3O2S: 485.0731, found: 486.0797 (M+H)

Example 96: (V-[(5/i_a)-5-(3-bromo-2-methylphenyl)-6-ethylthieno|2,3-iZ]pyriniÎdin-4yl]-£>-phenylalanine and

Example 97: .5-1(55//)-5-(3-010111 o-2-rnethylphenyl)-6-ethylthieno| 2,3-r/]pyriniidin-4ylJ-D-phenylalanine

Using General Procedure lia and Préparation 3a as the appropriate 5-iodo-thieno[2,3-i/| pyrimidine dérivative and Préparation 137 as the appropriate boronic acid dérivative, a mixture of diastereoisomers was obtained. They were separated via preparative reversed phase chromatography using 40 mM aqueous NH₄OAc solution (pH = 4, adjusted with

ORIGINAL

- 106AcOH) and acetonitrile as eluents. Example 96 was obtained as the earlier eluting diastereoisomer. HRMS calculated for Ci-iH^BrNjOiS: 495.0616, found: 496.0673 (M+H) Example 97 was obtained as the iater eluting diastereoisomer. HRMS calculated for C^H^BrNjOiS: 495.0616, found: 496.0687 (M+H)

Example 98:7V-[6-ethyl-5-(l/7-indazoI-4-yl)thieno[2,3-rf]pyrimidin-4-yl)-Z>phenylalanine

Using General Procedure Ha and Préparation 3a as the appropriate 5-iodo-thieno[2,3-c/] pyrimidine dérivative and 17y-indazol-4-ylboronic acid as the appropriate boronic acid dérivative, then purifying the crude product vîapreparative reversed phase chromatography using 0.1 % aqueous TFA solution and acetonitrile as eluents, gave Example 98 as a mixture of diastereoisomers. HRMS calculated for C24H21N5O2S: 443.1416, found: 444.1485 and 444.1481 (M+H)

Example 99: 7V-[6-ethyl-5-(quinolin-8-yl)thieno[2,3-rflpynmidin-4-yl]-Z>phenylalanine

Using General Procedure Ild and Préparation 3a as the appropriate 5-iodo-thieno[2,3-<:/] pyrimidine derivative and 8-quinolylboronic acid as the appropriate boronic acid dérivative, Examplc 99 was obtained as a mixture of diastereoisomers. HRMS calculated for C2₍₁H28N₄O₂S: 454.1463, found: 455.1558 (M+H)

Example 100: 7V-|6-ethyl-(5Æ_a)-5-(naphthalen-l-yi)thieno|2,3-i/lpyrimidin-4-yl|-Z)phenylalanine and

Example 101: jV-[6-cthyl-(55„)-5-(naphthalen-l-yl)thieno[2_t3-rf|pyrimidin-4-yl|-Z)phenylalanine

Separating the diastereoisomers of Example 79, using préparative reversed phase chromatography using 0.1 % aqueous TFA solution and acetonitrile as eluents,

ORIGINAL

- 107Example 100 was obtained as the earlier eluting diastereoisomer. HRMS calculated for HRMS calculated for C27H23N3O7S: 453.1511, found: 454.1596 (M+H)

Example 101 was obtained as the later eluting diastereoisomer. HRMS calculated for HRMS calculated for C27H23N3O2S: 453.1511, found: 454.1577 (M+H)

Example 102: 7V-[6-ethenyl-(5S_û)-5-(naphthalen-l-yl)thieno|2,3-rf|pyrimidin-4-yI]-Z>phenylalanine and

Example 103: A'-|6-ethenyl-(5/?„)-5-(naphthalen-l-yl)thieno|2,3-(/]pyriinidin--4-vl]-L>phenylalanine

Using General Procedure la and Préparation 4w as the appropriate 4-chloro-thieno[2,3-J] pyrimidine dérivative and Ώ-phenyIalanine as the appropriate amino acid dérivative. Example 102 was obtained as lhe earlier eluting diastereoisomer. HRMS calculated for C27H21N3O2S: 451.1354, found: 452.1411 (M+H)

Example 103 was obtained as the later eluting diastereoisomer. HRMS calculated for C₂7H₂₁N3O₂S: 451.1354, found: 452.1412 (M+H)

Example 104: /V-[(5S„)-5-(naphthalen-l-yl)-6-((lZ)-prop-l-en-l-yl)thieno[2,3-i7] pyrimidin-4-yl]-Z)-phenyIaIanine and

Example 105: A^f-[(57?„)-5-(naphthalen-l-yl)-6-((lZ)-prop-l-en-l-yI)thieno[2,3-i/| pyrimidin-4-yl]-Z>-phenylalanîne

Using General Procedure Ib and Préparation 4x as the appropriate 4-chloro-thieno[2,3-iZ] pyrimidine dérivative and £>-phenylalanine as the appropriate amino acid dérivative, a mixture of diastereoisomers was obtained. They were separated via préparative reversed phase chromatography using 40 mM aqueous NH4OAC solution (pH = 4, adjusted with AcOH) and acetonitriie as eluents. Example 104 was obtained as the earlier eluting diastereoisomer. HRMS calculated for C28H23N3O2S: 465.1511, found: 466.1577 (M+H) Example 104 also contains 55 % of Example 108.

ORIGINAL

- 108Example 105 was obtained as the later eluting diastereoisomer. HRMS calculated for C28H23N3O7S: 465.1511, found: 466.1578 (M+H)

Example 105 also contains 55 % Example 109.

Example 106: jV-[(55_e)-5-(naphthalen-l-yl)-6-(prop-l-en-2-yl)thieno[2,3-i/]pyrimidin4-ylJ-D-phenylalanine and

Example 107: jV-[(5R_fl)-5-(naphthalen-l-yl)-6-(prop-l-en-2-yI)thieno[2,3-rf]pyrimidin4-yl]-Z>-phenylalanine

Using General Procedure la and Préparation 4y as the appropriate 4-chloro-thieno[2,3-if] pyrimidine dérivative and D-phenylalanine as the appropriate amino acid dérivative, a mixture of diastereoisomers was obtained. They were separated via préparative reversed phase chromatography using 40 mM aqueous NH₄OAc solution (pH - 4, adjusted with AcOH) and acetonitrile as eluents. Example 106 was obtained as the earlier eluting diastereoisomer. HRMS calculated for C28H23N3O2S: 465.1511, found: 466.1581 (M+H) Example 107 was obtained as the later eluting diastereoisomer. HRMS calculated for C2sH23N3O₂S: 465.1511, found: 466.1597 (M+H)

Example 108: jV-fÎS^J-S-tnaphthalen-l-ylj-ô-Kli^-prop-l-en-l-ylItliienop^-i/l pyrimidin-4-yl}-Z)-phenylalanine and

Exaniple 109: /V-{(5Æ_fl)-5-(naphthalen-l-yl)-6-|(l£)-prop-l-en-l-yl]thieno|2,3-r/] pyrimidin-4-yl}-D-phenylaIanine

Using General Procedure la and Préparation 4z as the appropriate 4-chloro-thieno[2,3-i/] pyrimidine dérivative and £>-phenylalanine as the appropriate amino acid dérivative, a mixture of diastereoisomers was obtained. They were separated via préparative reversed phase chromatography using 40 mM aqueous NH4OAC solution (pH = 4, adjusted with AcOH) and acetonitrile as eluents. Example 108 was obtained as the earlier eluting diastereoisomer. HRMS calculated for C28H23N3O2S: 465.1511, found: 466.1593 (M+H)

ORIGINAL

- 109 Example 109 was obtained as the later eluting diastereoisomer. HRMS calculated for C28H23N3O2S: 465.1511, found: 466.1581 (M+H)

Example 110: N-[5-(3-chloro-2-methylphenyI)-6-ethylthieno[2,3-i/]pyrÎinidin-4-yl]-3( l//-pyrazol-lyl)alanine

Using General Procedure le and Préparation 4j as the appropriate 4-chloro-thieno[2,3-i7] pyrimidine dérivative and methyl 2-amino-3-pyrazol-l-yl-propanoate hydrochloride as the appropriate amino acid dérivative, then hydrolyzing the formed intermediate according to General Procedure VII, a mixture of diastereoisomers was obtained. It was purified via preparative reversed phase chromatography using 0.1 % aqueous TFA solution and acetonitrile as eluents, and Example 110 was obtained as the mixture of diastereoisomers. HRMS calculated for C_2IH₂oC1N₅0₂S: 441.1026, found: 442.1120 and 442.1123 (M+H)

Example 111: 7V-J(5Æ_n)-5-(3-chloro-2-methylphenyl)-6-ethyIthieno[2,3-if|pyrimidin-4yl]-3-cyclopentyl-Z>-alanine and

Example 112: jV-|(5S_n)-5-(3-chloro-2-mcthylphenyl)-6-ethylthieno[2,3-i/|pyrimidin-4yl]-3-cyclopentyl-Z>-aIanine

Using General Procedure Ib and Préparation 4j as the appropriate 4-chloro-thieno[2,3-i/] pyrimidine dérivative and (2Æ)-2-amino-3-cyclopentyl-propanoic acid as the appropriate amino acid dérivative, a mixture of diastereoisomers was obtained. They were separated via preparative reversed phase chromatography using 0.1 % aqueous TFA solution and acetonitrile as eluents. Example 111 was obtained as the earlier eluting diastereoisomer. HRMS calculated for C23H26CIN3O2S: 443.1434, found: 444.1519 (M+H)

Example 112 was obtained as the later eluting diastereoisomer. HRMS calculated for C₂3H26C1N₃O₂S: 443.1434, found: 444.1518 (M+H)

Example 113: Aⁱ-[(55’_fl)-5-(3-chIoro-2-methylphenyl)-6-ethylthieno[2,3-i/]pyrimidin-4yI]-£)-phenylalanine and

ORIGINAL

- I ΙΟ Example 114: j'V-KS/Gj-S-CS-chloro^-methylphenylj-ô-ethylthieno^S-i/JpyriniÎdin-dyl]-O-phenylalanine

Using General Procedure lia and Préparation 3a as the appropriate 5-iodo-thieno[2,3-i7] pyrimidine dérivative and (3-chloro-2-methyl-phenyl)boronic acid as the appropriate boronic acid dérivative, a mixture of diastereoisomers was obtained. They were separated via preparative reversed phase chromatography using 40 mM aqueous NFI₄OAc solution (pH = 4, adjusted with AcOH) and acetonitrile as eluents. Example 113 was obtained as the earlier eluting diastereoisomer. HRMS calculated for C24H22CIN3O2S: 451.1121. found: 452.1192 (M+H)

Example 114 was obtained as the later eluting diastereoisomer. HRMS calculated for C2.1H22CIN3O2S: 451.1121, found: 452.1174 (M+H)

Example 115: jV-[(5S„)-5-(3-chloro-2-niethylphenyl)-6-ethyltliieno|2,3-rf]pyrimidin-4yl|-Z.-phenylalanine and

Example 116: /V-[(5Æ_o)-5-(3-chloro-2-methylphenyl)-6-ethylthieno[2,3-rfjpyrimidin-4yl]-£-phenylalanine

Using General Procedure Ib and Préparation 4j as the appropriate 4-chloro-thieno[2,3-<:/] pyrimidine dérivative and Z-phenylalanine as the appropriate amino acid dérivative, a mixture of diastereoisomers was obtained. They were separated via preparative reversed phase chromatography using 0.1 % aqueous TFA solution and acetonitrile as eluents. Example 115 was obtained as the earlier eluting diastereoisomer. HRMS calculated for C24H22CIN3O2S: 451.1121, found: 452.1207 (M+H)

Example 116 was obtained as the later eluting diastereoisomer. HRMS calculated for C24H22CIN3O2S: 451.1121, found: 452.1183 (M+H)

Example 117: jV-|(57f_fl)-5-(3-diloro-2-methylphenyl)-6-ethylthieno[2,3-i/|pyrimidin-4yl[-3-cyclohexyl-/)-alanine and

ORIGINAL

- III Example 118: A/-[(55’_(J)-5-(3-chloro-2-methylphcnyl)-6-ethylthieno[2,3-rf]pyrimîdin-4yl]-3-cyclohexyl-Z>-aIanine

Using General Procedure 1b and Préparation 4j as the appropriate 4-chloro-thieno[2,3-<7] pyrimidine dérivative and (2Æ)-2-amino-3-cyclohexyl-propanoic acid as the appropriate amino acid dérivative, a mixture of diastereoisomers was obtained. They were separated via preparative reversed phase chromatography using 0.1 % aqueous TFA solution and acetonitrile as eluents. Example 117 was obtained as the earlier eluting diastereoisomer. HRMS calculated for C24H28CIN3O2S: 457.1591, found: 458.1672 (M+H)

Example 118 was obtained as the later eluting diastereoisomer. HRMS calculated for C24H28CIN3O2S: 457.1591, found: 458.1663 (M+H)

Example 119: 7V-[5-(3-chloro-2-methylphenyl)-6-ethylthieno [2,3-rT] pyrimîdin-4-y 1|alpha-methyl-Z>-phenylalanine

Using General Procedure Ib and Préparation 4j as the appropriate 4-chloro-thieno[2,3-c/] pyrimidine dérivative and (2Â)-2-amino-2-methyl-3-phenyl-propanoic acid as the appropriate amino acid dérivative, a mixture of diastereoisomers was obtained. It was purified via preparative reversed phase chromatography using 0.1 % aqueous TFA solution and acetonitrile as eluents. Example 119 was obtained as a mixture of diastereoisomers. HRMS calculated for C25H24CIN3O2S: 465.1278, found: 466.1372 and 466.1356 (M+H)

Example 120: A'-|(5/?_z,)-5-(3-chloro-2-methviphenyl)-6-ethylthieno[2,3-i^r/]pyriinidin-4yl]-2-hydroxy-Z)-phenylalanine and

Example 121; A^r-[(55^, _a)-5-(3-chloro-2-methy[phenyl)-6-ethylthieno[2,3-r/]pyrimidin-4yl|-2-hydroxy-£>-phenylaIanine

Using General Procedure Ib, Préparation 4j as the appropriate 4-chloro-thieno[2,3-iZ] pyrimidine dérivative, and (2J?)-2-amino-3-(2-hydroxyphenyl)propanoic acid as the appropriate amino acid dérivative, a mixture of diastereoisomers was obtained. They were separated via preparative reversed phase chromatography using 0.1 % aqueous TFA

ORIGINAL

- Il2solution and acetonitrile as eluents. Example 120 was obtained as the earlier eluting diastereoisomer. HRMS calculated for C24H22CIN3O3S: 467.1070, found: 468.1135 (M+H) Example 121 was obtained as the later eluting diastereoisomer. HRMS calculated for C24H22CIN3O3S: 467.1070, found: 468.1162 (M+H)

Example 122: (p5)-7V-[(57f_u)-5-(3-chloro-2-methylphenyl)-6-ethylthieno[2,3-rf| pyrimidin-4-yl]-beta-hydroxy-Z)-phenylalanine and

Example 123: (p_lS)-A^f-|(5«S_a)-5-(3-chloro-2-methylphenyl)-6-ethylthieno[2,3-/Z| pyrimidin-4-yl|-beta-hydroxy-Z>-phenylalanine

Using General Procedure Ib and Préparation 4j as the appropriate 4-chloro-thieno[2,3-t/] pyrimidine dérivative and (2Æ,3S)-3-phenylserine as the appropriate amino acid dérivative, a mixture of diastereoisomers was obtained. They were separated via préparative reversed phase chromatography using 0.1 % aqueous TFA solution and acetonitrile as eluents. Example 122 was obtained as the earlier eluting diastereoisomer. HRMS calculated for C24H22CIN3O3S; 467.1070, found: 468.1151 (M+H)

Example 123 was obtained as the later eluting diastereoisomer. HRMS calculated for C₂4H₂2CIN3O₃S: 467.1070, found: 468.1133 (M+H)

Example 124: (p7f)-/V-[(5/f_o)-5-(3-chloro-2-inetbylphenyi)-6-ethylthieno[2,3-i/| pyrimidin-4-yl]-beta-hydroxy-L-phenylalanine and

Example 125: (p/î)-7V-[(5S_i/)-5-(3“chloro-2-methylphcnyl)-6-ethylthieno[2,3-i/| pyrimidin-4-yl]-bcta-hydroxy-L-phcnylalanine

Using General Procedure Ib and Préparation 4j as the appropriate 4-chloro-thieno[2,3-t/] pyrimidine dérivative and (2_tS’,3/f)-2-amino-3-hydroxy-3-phenylpropionic acid as the appropriate amino acid dérivative, a mixture of diastereoisomers was obtained. They were separated via préparative reversed phase chromatography using 0.1 % aqueous TFA solution and acetonitrile as eluents. Example 124 was obtained as the earlier eluting diastereoisomer. HRMS calculated for C24H22CIN3O3S: 467.1070, found: 468.1144 (M+H)

ORIGINAL

- II3Example 125 was obtained as the later eluting diastereoisomer. HRMS calculated for C24H22CIN3O3S: 467.1070, found: 468.1153 (M+H)

Example 126: A^z-[(5/i_(I)-5-(3-chloro-2-methylphenyl)-6-ethylthieno[2,3-iZ]pyrimidin-4yl]-2-cyano-Z)-phenylalanine and

Example 127 jV-|(55'„)-5-(3-chloro-2-methylphenyl)-6-ethylthieno[2,3-i/]pyrimidin-4yl]-2-cyano-D-phenylalanine

Using General Procedure lb and Préparation 4j as the appropriate 4-chloro-thieno[2,3-i7] pyrimidine dérivative and (2Æ)-2-amino-3-(2-cyanophenyl)propanoic acid as the appropriate amino acid dérivative, a mixture of diastereoisomers was obtained. They were separated via préparative reversed phase chromatography using 0.1 % aqueous TFA solution and acetonitrile as eluents. Example 126 was obtained as the earlier eluting diastereoisomer. HRMS calculated for C25H21CIN4O2S: 476.1074, found: 477.1129 (M+H) Example 127 was obtained as the later eluting diastereoisomer. HRMS calculated for C25H21CIN4O2S: 476.1074, found: 477.1134 (M+H)

Example 128: A-[(57?_(J)-5-(3-chloro-2-methylphcnyl)-6-cthylthieno[2,3-i/]pyrinudin-4yl]-2-methoxy-Z)-phenylalanine and

Example 129: 7V-|(5₁S'₍,)-5-(3-chIoro-2-methylphenyl)-6-ethylthieno[2,3-i/]pyrimidin-4yl]-2-methoxy-2)-phenj (alanine

Using General procedure le and Préparation 4j as the appropriate 4-chloro-thîeno[2,3-t/] pyrimidine dérivative and (2A)-2-amino-3-(2-inethoxyphenyl)propanoic acid as the appropriate amino acid dérivative, a mixture of diastereoisomers was obtained. They were separated via préparative reversed phase chromatography using 0.1 % aqueous TFA solution and acetonitrile as eluents. Example 128 was obtained as the earlier eluting diastereoisomer. HRMS calculated for C25H24CIN3O3S: 481.1227, found: 482.1320 (M+H) Example 129 was obtained as the later eluting diastereoisomer. HRMS calculated for C25H24CIN3O3S: 481.1227, found: 482.1319 (M+H)

ORIGINAL

- Il4Example 130: 7V-|(57f_ff)-5-(3-chloro-2-methylphenyl)-6-ethylthieno[2,3-i/]pyrimidin-4yl]-2,6-difluoro-Z)-phenylalanine and

Example 131: /V-f(5S'_a)-5-(3-chloro-2-methylplienyl)-6-ethylthieno|2,3-//]pynniidin-4yl]-2,6-difluoro-Z)-phenyIalanine

Using General Procedure Ib and Préparation 4j as the appropriate 4-chloro-thieno[2,3-i/] pyrimidine dérivative and (2Æ)-2-amino-3-(2,6-difluorophenyl)propanoic acid as the appropriate amino acid dérivative, a mixture of diastereoisomers was obtained. They were separated via préparative reversed phase chromatography using 0.1 % aqueous TFA solution and acetonitrile as eluents. Example 130 was obtained as the earlier eluting diastereoisomer. HRMS calculated for C24H20CIF2N3O2S: 487.0933, found: 488.1009 (M+H)

Example 131 was obtained as the later eluting diastereoisomer. HRMS calculated for C24H20CIF2N3O2S: 487.0933, found: 488.1020 (M+H)

Example 132: (2Æ)-2-{|(5/f_(i)-5-(3-chloro-2-methylphenyl)-6-ethylthieno[2,3-i/] pyrimidin-4-yl]amino}-3-(l//-indol-4-yI)propanoic acid and

Example 133: (2/?)-2-{|(5S„)-5-(3-chloro-2-methyIphenyl)-6-ethylthieno[2,3-rf| pyrimidin-4-yl|amino}-3-(lff-indol-4-yI)propanoic acid

Using General Procedure Ib and Préparation 4j as the appropriate 4-chloro-thieno[2,3-c/| pyrimidine dérivative and (2Æ)-2-amino-3-(l//-indoL4-yl)propanoic acid hydrochloride as the appropriate amino acid dérivative, a mixture of diastereoisomers was obtained. They were separated via preparative reversed phase chromatography using 0.1 % aqueous TFA solution and acetonitrile as eluents. Example 132 was obtained as the earlier eluting diastereoisomer. HRMS calculated for C26H23CIN4O2S: 490.1230, found: 491.1289 (M+H) Example 133 was obtained as the later eluting diastereoisomer. HRMS calculated for C₂₆H23CtN₄O₂S: 490.1230, found: 491.1309 (M+H)

ORIGINAL

-usExample 134: 2-carbamoyI-JV-[(55_ff)-5-(3-chloro-2-methylphenyl)-6-ethylthieno[2,3-rf] pyrimidin-4-yl]-Z>-phenylalanine

Using General Procedure Ib and Préparation 4j as the appropriate 4-chloro-thieno[2,3-i7] pyrimidine dérivative and (2/?)-2-amino-3-(2-carbamoylphenyl)propanoic acid as the appropriate amino acid dérivative, a mixture of diastereoisomers was obtained. They were separated via préparative reversed phase chromatography using 0.1 % aqueous TFA solution and acetonitrile as eluents. Example 134 was obtained as the later eluting diastereoisomer. HRMS calculated for CjsH^.iCINjOjS: 494.1179, found: 495.1255 (M+H)

Example 135: /V-|(5Æ„)-5-(3-chloro-2-methylphenyl)-6-etliyIthieno|2,3-r/]pyrimidin-4yl]-2-nitro-Z>-phenylalanine and

Example 136: A^r-|(55_u)-5-(3-chloro-2-methylphenyI)-6-ethylthieno|2,3-rf]pyrimidin-4yI]-2-nitro-Z)-phenylalanine

Using General Procedure 1b and Préparation 4j as the appropriate 4-chloro-thieno[2,3-ff] pyrimidine dérivative and (2Æ)-2-amino-3-(2-nitrophenyl)propanoic acid as the appropriate amino acid dérivative, a mixture of diastereoisomers was obtained. They were separated via préparative reversed phase chromatography using 0.1 % aqueous TFA solution and acetonitrile as eluents. Example 135 was obtained as the earlier eluting diastereoisomer. HRMS calculated for C24H21CIN4O4S: 496.0972, found: 497.1026 (M+H)

Example 136 was obtained as the later eluting diastereoisomer. HRMS calculated for C24H21CIN4O4S: 496.0972, found: 497.1045 (M+H)

Example 137: 7V-|(5.ff_fl)-5-(3-chloro-2-inethylphenyl)-6-ethylthieno[2,3-i/]pyrimidîn-4~ yl|-2-(trifluoromethyl)-Z>-phenylaIanine and

Example 138: A^r-|(5S'„)-5-(3-chloro-2-methylphenyl)-6-ethylthieno[2,3-r/|pyrimidin-4yl]-2-(trifluoromethyl)-£>-phenylalanine

ORIGINAL

- Il6Using General Procedure Ib and Préparation 4j as the appropriate 4-chloro-thieno[2,3-<Y] pyrimidine derivative and (2/?)-2-amino-3-[2-(trifluoromethyl)phenyl]propanoic acid as the appropriate amino acid derivative, a mixture of diastereoisomers was obtained. They were separated via préparative reversed phase chromatography using O.l % aqueous TFA solution and acetonitrile as eluents. Example 137 was obtained as the earlier eluting diastereoisomer. HRMS calculated for C25H21CIF3N3O2S: 519.0995, found: 520.1068 (M+H)

Example 138 was obtained as the later eluting diastereoisomer. HRMS calculated for C25H21CIF3N3O2S: 519.0995, found: 520.1047 (M+H)

Example 139: 2-bromo-/V-[(5Æ_i,)-5-(3-chloro-2-methylphenyI)-6-ethylthicno[2,3-r/| pyrimidin-4-yl]-Z)-phenylaIanine and

Example 140: 2-bromo-A'-[(5S'_fl)-5-(3-chloro-2-metliylphenyl)-6-ethylthieno[2,3-<7] pyrimidin-4-yl|-D-phenylalanine

Using General Procedure Ib and Préparation 4j as the appropriate 4-chloro-thieno[2,3-i/] pyrimidine derivative and (27?)-2-amino-3-[2-bromophenyl]propanoic acid as the appropriate amino acid derivative, a mixture of diastereoisomers was obtained. They were separated via préparative reversed phase chromatography using 0.1 % aqueous TFA solution and acetonitrile as eluents. Example 139 was obtained as the earlier eluting diastereoisomer. HRMS calculated for C24H2iClBrN₃O₂S: 529.0226, found: 530.0312 (M+H)

Example 140 was obtained as the later eluting diastereoisomer. HRMS calculated for C24H₂]ClBrN₃O2S: 529.0226, found: 530.0294 (M+H)

Example 141 : /V-[(57?_a)-5-(3-chIoro-2-inetliylphenyI)-6-ethylthieno[2,3-i/]pyrimi(lin-4yl|-2-[2-(dimethylamino)-2-oxoethoxy|-Z>-phenylalanine and

Example 142: A'-[(55j-5-(3-chl<no-2-methylphenvl)-6-ethylthieno|2,3-i/[pyrnnidin-4yl|-2-|2-(dimethylamino)-2-oxocthoxy]-Z)-phenylalanine

ORIGINAL

- 117Using General Procedure Ib and Préparation 4j as the appropriate 4-chioro-thieno[2,3-i/] pyrimidine dérivative and Préparation A9 as the appropriate amino acid dérivative, a mixture of diastereoisomers was obtained. They were separated via préparative reversed phase chromatography using O.l % aqueous TFA solution and acetonitrile as eluents. 5 Example 141 was obtained as the earlier eluting dîastereoisomer. HRMS calculated for

C28H29CIN4O4S: 552.1598, found; 553.1694 (M+H)

Example 142 was obtained as the later eluting dîastereoisomer. HRMS calculated for C₂₈H₂₉CIN4O4S: 552.1598, found: 553.1673 (M+H)

Example 143: W-|(5Z?_a)-5-(3-chk>ro-2-methylphenyl)-6-ethylthieno|23-'/|pyrimidin-410 y l]-2-(2-cyclopenty Iethoxy)-£*-pheny lalanine and

Example 144: A^f-((55_a)-5-(3-chloro-2-methylphenyl)-6-ethylthieno|2,3-i/]pyrimidin-4yl]-2-(2-cycIopentj’lethoxy)-Z)-phcnylalanine

Using General Procedure Ib and Préparation 4j as the appropriate 4-chloro-thieno[2,3-i/] 15 pyrimidine dérivative and Préparation A10 as the appropriate amino acid dérivative, a mixture of diastereoisomers was obtained. They were separated via préparative reversed phase chromatography using 0.1 % aqueous TFA solution and acetonitrile as eluents. Example 143 was obtained as the earlier eluting dîastereoisomer. HRMS calculated for C31H34CIN3O3S: 563.2009, found: 564.2106 (M+H)

Example 144 was obtained as the later eluting dîastereoisomer. HRMS calculated for C31H34CIN3O3S: 563.2009, found: 564.2101 (M+H)

Examplc 145: 7V-[(5Æ_u)-5-(3-chloro-2-mcthylphenyI)-6-ethylthieno[2,3-i/]pyrimidin-4yl]-2-(2-phenylethoxy)-Z>-phenylalanine and

Example 146: 7V-[(55_(I)-5-(3-chloro-2-methyIphenyl)-6-ethylthieno|2,3-i/|pyrimidin-4yl|-2-(2-phenylethoxy)-Z>-phenylalanine

Using General Procedure Ib and Préparation 4j as the appropriate 4-chloro-thieno[2,3-iZ] pyrimidine dérivative and Préparation Al 1 as the appropriate amino acid dérivative, a

ORIGINAL

- usmixture of diastereoisomers was obtained. They were separated via préparative reversed phase chromatography using 0.1 % aqueous TFA solution and acetonitrile as eluents. Example 145 was obtained as the earlier eluting diastereoisomer. HRMS calculated for C32H30CIN3O3S: 571.1696, found: 572.1769 (M+H)

Example 146 was obtained as the later eluting diastereoisomer. HRMS calculated for C32H30CIN3O3S: 571.1696, found: 572.1763 (M+H)

Example 147: /V-[(57î_a)-5-(3-chloro-2-methylphenyl)-6-ethylthieno[2,3-i/]pyrimidin-4yI]-2-(3-phenylpropoxy)-Z)-phenylalanine and

Example 148: /V-|(5.S’„)-5-(3-ehloro-2-niethylphenyl)-6-ethylthieno|2,3-i/|pyriniidin-4yl]-2-(3-phenylpropoxy)-D-phenylalanine

Using General Procedure Ib and Préparation 4j as the appropriate 4-chloro-thieno[2,3-rf] pyrimidine dérivative and Préparation Al2 as the appropriate amino acid dérivative, a mixture of diastereoisomers was obtained. They were separated via préparative reversed phase chromatography using 0.1 % aqueous TFA solution and acetonitrile as eluents. Example 147 was obtained as the earlier eluting diastereoisomer. HRMS calculated for C33H32CIN3O3S: 585.1853, found: 586.1917 (M+H)

Example 148 was obtained as the later eluting diastereoisomer. HRMS calculated for C33H32CIN3O3S: 585.1853, found: 586.1906 (M+H)

Example 149: 2-[(3-chlorobenzyl)oxy]-/V-[(5/f_a)-5-(3-chloro-2-methylphenyl)-6-ethyl thieno|2,3-rf]pyrimidin-4-ylJ-Z)-phenylalanme and

Example 150: 2-|(3-chlorobenzyl)oxy]-7V-|(5S’_a)-5-(3-chloro-2-niethylphenyl)-6-ethyl thieno [2,3-(7] pyrimidin-4-yl]-D-phenylalanine

Using General Procedure Ib and Préparation 4j as the appropriate 4-chloro-thieno[2,3-t/| pyrimidine dérivative and Préparation A13 as the appropriate amino acid dérivative, a mixture of diastereoisomers was obtained. They were separated via préparative reversed phase chromatography using 0.1 % aqueous TFA solution and acetonitrile as eluents.

ORIGINAL

- II9Example 149 was obtained as the earlier eluting diastereoisomer. HRMS calculated for C31H27CI2N3O3S: 591.1150, found: 592.1211 (M+H)

Example 150 was obtained as the later eluting diastereoisomer. HRMS calculated for C31H27CI2N3O3S: 591.1150, found: 592.1234 (M+H)

Example 151: jV-[(57f_fl)-5-(3-chloro-2-methylphenyl)-6-ethylthieno[2,3-7]pyrimidin-4yl|-3-pyridin-2-yl-D-alamne and

Example 152: A^r-|(5_lS_a)-5-(3-chloro-2-methylphenyl)-6-ethyithieno|2,3-7|pyrimidin-4yl]-3-pyridin-2-yl-Z>-alanine

Using General Procedure Ib and Préparation 4j as the appropriate 4-chloro-thieno[2,3-7J pyrimidîne dérivative and (2/?)-2-amino-3-(2-pyridyi)propanoic acid as the appropriate amino acid dérivative, a mixture of diastereoisomers was obtained. They were separated via preparative reversed phase chromatography using 0.1 % aqueous TFA solution and acetonitrile as eluents. Example 151 was obtained as the earlier eluting diastereoisomer. HRMS calculated for C₂3H₂|C1N₄O₂S: 452.1074, found: 453.1146 (M+H)

Example 152 was obtained as the later eluting diastereoisomer. HRMS calculated for C₂3H2iCIN₄O₂S: 452.1074, found: 453.1135 (M+H)

Example 153: A/-|(57Î₀)-5-(3-cliloro-2-methylpheiiyI)-6-ethylthieno|2,3-7|pynmidin-4yl]-2-[2-(4-methyIpipcrazin-l-yl)ethoxy]-Z>-phenylalanine and

Exaniple 154: 7V-|(55’₍,)-5-(3-chloro-2-methylphenyI)-6-etliylthieno[2,3-r/]pyrimidin-4yI]-2-[2-(4-methyIpiperazin-l-yl)ethoxy]-Z)-phenylalaninc

Using General Procedure Ib and Préparation 4j as the appropriate 4-chloro-thieno[2,3-7] pyrimidîne dérivative and Préparation A14 as the appropriate amino acid dérivative, a mixture of diastereoisomers was obtained. They were separated via preparative reversed phase chromatography using 0.1 % aqueous TFA solution and acetonitrile as eluents. Example 153 was obtained as the earlier eluting diastereoisomer. HRMS calculated for C31H36CIN5O3S: 593.2227, found: 594.2297 (M+H)

ORIGINAL

- 120Example 154 was obtained as the later eluting diastereoisomer. HRMS calculated for C₃iH₃₆C1N₅O₃S: 593.2227, found: 594.2289 (M+H)

Example 155: A^r-[(5Æ_a)-5-(3-chloro-2-methylphenyl)-6-ethylthieno[2,3-r/|pyrimidin-4yl]-2-[2-(dimethylamino)ethoxy]-Z)-phenylalanine and

Example 156: A-[(5S_fl)-5-(3-cliloro-2-methylphenyI)-6-etliylthieno|2,3-i/]pyrimidin-4yl|-2-|2-(dimethylamino)ethoxy]-£)-phenylaIanine

Using General Procedure Ib and Préparation 4j as the appropriate 4-chloro-thieno[2,3-cZ] pyrimidine dérivative and Préparation Al5 as the appropriate amino acid dérivative, a mixture of diastereoisomers was obtained. They were separated via préparative reversed phase chromatography using 0.1 % aqueous TFA solution and acetonitrile as eluents. Example 155 was obtained as the earlier eluting diastereoisomer. HRMS calculated for C2sH₃iC1N₄O3S: 538.1805, found: 539.1890 (M+H)

Example 156 was obtained as the later eluting diastereoisomer. HRMS calculated for C28H_3IC1N₄O₃S; 538.1805, found: 539.1887 (M+H)

Example 157: A^r-|(57f_<ï)-5-(3-chloro-2-methylphenyl)-6-ethylthieno|2,3-i/|pyrimidin-4yl]-2-[3-(dimethylamino)propoxy]-Z)-phenylalanine and

Example 158: /V-[(5S’_u)-5-(3-chloro-2-methylphenyl)-6-ethylthieno|2,3-J]pyrimidin-4yl|-2-|3-(dimethylamino)propoxy|-/)-phenylalanine

Using General Procedure Ib and Préparation 4j as the appropriate 4-chloro-thienof2,3-c/] pyrimidine dérivative and Préparation Al6 as lhe appropriate amino acid dérivative, a mixture of diastereoisomers was obtained. They were separated via préparative reversed phase chromatography using 0.1 % aqueous TFA solution and acetonitrile as eluents. Example 157 was obtained as the earlier eluting diastereoisomer. HRMS calculated for C29H33CIN4O3S: 552.1962, found: 553.2043 (M+H)

Example 158 was obtained as the later eluting diastereoisomer. HRMS calculated for C₂9H₃₃C1N₄O₃S: 552.1962, found: 553.2053 (M+H).

ORIGINAL

- I2l Example 159: 3-cyclopropyl-7V-[6-ethyl-5-(naphthalen-l-yl)thieno[2,3-rf|pyrimidin-4yl]-Z)-alanine

Using General Procedure Ib and Préparation 4k as the appropriate 4-chloro-thieno[2,3-i/] pyrimidine dérivative and (27î)-2-amîno-3-cyclopropyl-propanoic acid as the appropriate amino acid dérivative, a mixture of diastereoisomers was obtained. They were separated via préparative reversed phase chromatography using 0.1 % aqueous TFA solution and acetonitrile as eluents. Example 159 was obtained as the mixture of diastereoisomers. HRMS calculated for C24H23N3O2S: 417.1511, found: 418.1570 (M+H)

Example 160: (2Æ)-{[6-ethyl-5-(naphthalen-l-yI)thieno[2,3-J]pyrimidin-4-yl|amino} (phenyl)ethanoic acid, diastereoisomer 1 and

Example 161 : (27î)-{[6-ethyl-5-(naphthalen-l-yl)th!eno[2,3-i/]pyrimidin-4-yl]aniino} (phcnyl)ethanoic acid, diastereoisomer 2

Using General Procedure Ib and Préparation 4k as the appropriate 4-chloro-thieno[2,3-c/] pyrimidine dérivative and (2Æ)-2-amino-2-phenyl-acetic acid as the appropriate amino acid dérivative, using DMA as solvent instead of DMSO, a mixture of diastereoisomers was obtained. They were separated via préparative reversed phase chromatography using 0.1 % aqueous TFA solution and acetonitrile as eluents. Example 160 was obtained as the earlier eluting diastereoisomer. HRMS calculated for C26H21N3O2S: 439.1354, found: 440.1428 (M+H)

Example 161 was obtained as the later eluting diastereoisomer. HRMS calculated for C26H21N3O2S: 439.1354, found: 440.1412 (M+H)

Example 162: A-[6-cthyl-(55_ti)-5-(naphthalen-I-Yl)thieno|2,3-i/jpyrimidin-4-yI]-3pyridin-3-yl-Z>-alanine and

Example 163: V-|6-cthyl-(5A„)-5-(naphthalen-1-yl)thieno|2,3-i/]pyrimidin-4-yl|-3pyridin-3-yl-Z>-aIanine

ORIGINAL

- 122 Using General Procedure Ib and Préparation 4k as the appropriate 4-chloro-thieno[2,3-<7] pyrimidine dérivative and (27î)-2-amino-3-(3-pyridyl)propanoic acid as the appropriate amino acid dérivative, a mixture of diastereoisomers was obtained. They were separated via preparative reversed phase chromatography using 40 niM aqueous NH4OAC solution (pH = 4, adjusted with AcOH) and acetonitrile as eluents. Example 162 was obtained as the earlier eluting diastereoisomer. HRMS calculated for C26H22N4O2S: 454.1463, found: 455.1520 (M+H)

Example 163 was obtained as the later eluting diastereoisomer. HRMS calculated for C26H22N4O2S: 454.1463, found: 455.1536 (M+H)

Example 164: 3-cyclohexyl-7V-|6-ethyl-5-(naphthalen-l-yl)thieno[2,3-rf]pyrimi(!in-4yl]-Z)-alanine

Using General Procedure la and Préparation 4k as the appropriate 4-chloro-thieno[2,3-<7] pyrimidine dérivative and (27î)-2-amino-3-cyclohexyl-propanoic acid as the appropriate amino acid dérivative, a mixture of diastereoisomers was obtained. It was purified via preparative reversed phase chromatography using 0.02 % aqueous HCOOH solution and acetonitrile as eluents. Example 164 was obtained as the mixture of diastereoisomers. HRMS calculated for C27H29N3O2S: 459.1980, found: 460.2042 (M+H)

Example 165: 3-cyclohexyl-/V-[6-ethyl-(5Æ_fl)-5-(naphthalen-l-yl)thieno[2,3-i/| pyrimidin-4-yl]-£>-alanine and

Example 166: 3-cyclohexyl-jV-|6-etliyl-(5S_i;)-5-(naphthalen-l-yl)thieno|2,3-rf] pyrimidin-4-yl|-£>-aIanine

Diastereoisomers of Example 164 were separated via preparative reversed phase chromatography using 40 mM aqueous NH4OAC solution (pH = 4, adjusted with AcOH) and acetonitrile as eluents. Example 165 was obtained as the earlier eluting diastereoisomer. HRMS calculated for C27H29N3O2S: 459.1980, found: 460.2043 (M+H)

ORIGINAL

- 123 Example 166 was obtained as the later eluting diastereoisomer. HRMS calculated for C27H29N3O2S: 459.1980, found: 460.2058 (M+H)

Example 167: /V-|6-cthyl-5-(naphthalen-I-yl)thieno|2,3-r/]pyrimidin-4-yl|-2-methyl-Z)phenylalanine, diastereoisomer 1 and

Example 168: jV-[6-ethyl-5-(naphthalen-l-yl)thieno|2,3-i/]pyrimidin-4-yl]-2-methyl-Z)plienylalanine, diastereoisomer 2

Using General Procedure Ib and Préparation 4k as the appropriate 4-chloro-thieno[2,3-<7] pyrimidine dérivative and D-2’-methylphenylalanine as the appropriate amino acid derivative, a mixture of diastereoisomers was obtained. They were separated via préparative reversed phase chromatography using 40 mM aqueous NHjOAc solution (pH =4, adjusted with AcOH) and acetonitrile as eluents. Example 167 was obtained as the earlier eluting diastereoisomer. HRMS calculated for C28H25N3O2S: 467.1667, found: 468.1747 (M+H)

Example 168 was obtained as the later eluting diastereoisomer. HRMS calculated for C28H25N3O2S: 467.1667, found: 468.1748 (M+H)

Example 169: (2Æ)-2-{[6-ethyI-5-(naphthalen-l-yl)thieno|2,3-rfIpyrimidin-4-yI] amino}-4-phenylbutanoic acid

Using General Procedure la and Préparation 4k as the appropriate 4-chloro-thieno[2,3-c/] pyrimidine derivative and (2/?)-2-amino-4-phenyl-butanoic acid as the appropriate amino acid derivative, a mixture of diastereoisomers was obtained. Il was purified via préparative reversed phase chromatography using 0.02 % aqueous HCOOH solution and acetonitrile as eluents. Example 169 was obtained as mixture of diastereoisomers. HRMS calculated for C28H25N3O2S: 467.1667, found: 468.1731 (M+H)

Example 170: (2/?)-2-{|6-ethyl-(55„)-5-(naphthalen-l-yl)tliicno|2,3-rf]pyrimidin-4-yl| amino}-4-phenylbutanoic acid and

ORIGINAL

- I24Example 171: (27î)-2-{[6-ethyl-(57f_a)-5-(naphthalen-l-yl)thieno|2,3-rf|pyrimidin-4-yl| amino]—l-phenylbutanoic acid

Diastereoisomers of Example 169 were separated via préparative reversed phase chromatography using 40 mM aqueous NH₄OAc solution (pH = 4, adjusted with AcOH) and acetonitrile as eluents. Example 170 was obtained as the earlier eluting diastereoisomer. HRMS calculated for C28H25N3O2S: 467.1667, found: 468.1733 (M+H) Example 171 was obtained as the later eluting diastereoisomer. HRMS calculated for C28H25N3O2S: 467.1667, found: 468.1726 (M+H)

Example 172: jV-|6-ethyl-(5Æ„)-5-(naphtlialen-l-yl)thieno[2,3-i/]pyrimidin-4-yl]-Z)tyrosine and

Example 173: _JV-|6-ethyl-(55'_a)-5-(naphthalen-l-yl)thieno[2,3-rf]pyrimidin-4-yl]-Z)tyrosine

Using General Procedure Ib and Préparation 4k as the appropriate 4-chloro-thieno[2,3-i/] pyrimidine dérivative and (2/î)-2-amino-3-(4-hydroxyphenyl)propanoic acid as the appropriate amino acid dérivative, using DMA as solvent instead of DMSO, a mixture of diastereoisomers was obtained. They were separated via préparative reversed phase chromatography using 0.1 % aqueous TFA solution and acetonitrile as eluents. Example 172 was obtained as the earlier eluting diastereoisomer. HRMS calculated for C27H23N3O3S: 469.1460, found: 470.1539 (M+H)

Exampie 173 was obtained as the later eluting diastereoisomer. HRMS calculated for C27II23N3O3S: 469.1460, found: 470.1534 (M+H)

Example 174: A/-[6-ethyl-(5S_a)-5-(naphthalen-l-yl)thieno[2,3-i/]pyrimidin-4-yl]-2hydroxy-Z>-plienylalanine and

Example 175: AMô-ethyl-tSlî^-S-fnaphtlialen-l-yllthieno^^-i/Ipyrimidin^-yl]^hydroxy-D-phenylalanine

ORIGINAL

- 125 Using General Procedure Ib and Préparation 4k as the appropriate 4-chloro-thieno[2,3-if] pyrimidine dérivative and (2Æ)-2-amino-3-(2-hydroxyphenyl)propanoic acid as the appropriate amino acid dérivative, a mixture of diastereoisomers was obtained. They were separated via preparative reversed phase chromatography using 40 mM aqueous NH₄OAc solution (pH = 4, adjusted with AcOH) and acetonitrile as eluents. Example 174 was obtained as the earlier eiuting diastereoisomer. HRMS calculated for C27H23N3O3S: 469.1460. found: 470.1546 (M+H)

Example 175 was obtained as the later eiuting diastereoisomer. HRMS calculated for C27H2₃N₃O₃S: 469.1460, found: 470.1520 (M+H)

Example 176: A^/-[6-ethyl-(57?_i,)-5-(naphthalen-l-yI)tliieno[2,3-r/]pyriinidîn-4-yl]-4fluoro-D-phenylalanine and

Example 177: 7V-[6-ethyl-(55'_a)-5-(naphthaIen-l-yl)thieno[2,3-rf]pyrimidÎn-4-yl]-4fluoro-Z>-phenylalanine

Using General Procedure 1b and Préparation 4k as the appropriate 4-chloro-thieno[2,3-i7] pyrimidine dérivative and (2Æ)-2-amino-3-(4-fluorophenyl)propanoic acid as the appropriate amino acid derivatîve, using DMA as solvent instead of DMSO, a mixture of diastereoisomers was obtained. They were separated via preparative reversed phase chromatography using 0.1 % aqueous TFA solution and acetonitrile as eluents. Example 176 was obtained as the earlier eiuting diastereoisomer. HRMS calculated for C27H2₂FN₃O2S: 471.1417, found: 472.1493 (M+H)

Example 177 was obtained as the later eiuting diastereoisomer. HRMS calculated for C77H22FNÎO2S: 471.1417, found: 472.1494 (M+H)

Example 178: iV-[6-ethyl-(57f_fl)-5-(naphthalen-l-yl)thieno|2,3-r/]pyrimidin-4-yl|-3fluoro-D-phcnylalanine and

Example 179: 7V-|6-ethyl-(55^, _fl)-5-(naphtlialen-l-yl)thieno[2,3-rf]pyrimidin-4-yl]-3fluoro-Z>-phenylalanine

ORIGINAL

- 126Using General Procedure Ib and Préparation 4k as the appropriate 4-chloro-thieno[2,3-<7] pyrimidine dérivative and (27?)-2-amino-3-(3-fluorophenyl)propanoic acid as the appropriate amino acid dérivative, a mixture of diastereoisomers was obtained. They were separated via préparative reversed phase chromatography using O.l % aqueous TFA solution and acetonitrile as eluents. Example 178 was obtained as the earlier eluting diastereoisomer. HRMS calculated for C27H22FN3O2S: 471.1417, found: 472.1486 (M+H) Example 179 was obtained as the later eluting diastereoisomer. HRMS calculated for C27H22FN3O2S: 471.1417, found: 472.1482 (M+H)

Example 180: Λ-|6-υίΙιν1-(5/?,_;)-5-(ηηρ1ιί1ι;ι1εη-1-yl)thieno|2.3-7|pyriniidin-4-yl|-2fluoro-Z>-phenylalanine and

Example 181: iV-|6-ethyI-(55_fl)-5-(naphthalen-l-yl)thieno|2,3-rf]pyrimidin-4-yI]-2fluoro-D-phenylalanine

Using General Procedure Ib and Préparation 4k as the appropriate 4-chIoro-thieno[2,3-t/] pyrimidine dérivative and (27Î)-2-amino-3-(2-tluorophenyl)propanoic acid as the appropriate amino acid dérivative, using DMA as solvent instead of DMSO, a mixture of diastereoisomers was obtained. They were separated via préparative reversed phase chromatography using 0.1 % aqueous TFA solution and acetonitrile as eluents. Example 180 was obtained as the earlier eluting diastereoisomer. HRMS calculated for C27H22FN3O2S: 471.1417, found: 472.1501 (M+H)

Exaniple 181 was obtained as the later eluting diastereoisomer. HRMS calculated for C27H22FN3O2S: 471.1417, found: 472.1492 (M+H)

Example 182: A'-|6-ethyl-5-(naphthalen-l-yl)thieno[2,3-rf]pyrimidin-4-yl|-2-methoxyZ)-phcnylalanine

Using General Procedure Ib and Préparation 4k as the appropriate 4-chloro-thieno[2,3-i/] pyrimidine dérivative and (2Æ)-2-amino-3-(2-methoxyphenyl)propanoic acid as the appropriate amino acid dérivative, a mixture of diastereoisomers was obtained. It was purified via préparative reversed phase chromatography using 40 mM aqueous NHjOAc

ORIGINAL

-I27solution (pFI = 4, adjusted with AcOH) and acetonitrile as eluents. Example 182 was obtained as a mixture of diastereoisomers. HRMS calculated for C28H25N3O3S; 483.1617, found: 484.1682 and 484.1695 (M+H)

Example 183: 2-chloro-jV-[6-ethyl-5-(naphthalen-l-yl)thienof2,3-i/]pyrimi(IÎn-4-yI]-£)phenylalanine

Using General Procedure Ib and Préparation 4k as the appropriate 4-chloro-thieno[2,3-<7] pyrimidine dérivative and (2Æ)-2-amino-3-(2-chlorophenyl)propanoic acid as the appropriate amino acid dérivative, a mixture of diastereoisomers was obtained. It was purified via preparative reversed phase chromatography using 0.1 % aqueous TFA solution and acetonitrile as eluents. Example 183 was obtained as the mixture of diastereoisomers. HRMS calculated for C27H22CIN3O2S: 487.1121, found: 488.1198 and 488.1199 (M+H)

Example 184: 7V-[6-ethyl-(5ff_a)-5-(naphthalen-l-yl)thieno[2,3-rf]pyrimidin-4-yl]-Z>tryptophan and

Example 185: A-|6-etlivl-(5Xj-5-(naphthak'n-l-yI)thieno[2,3-<7|pYrimidin-4-yl]-Dtryptophan

Using General Procedure Ib and Préparation 4k as the appropriate 4-chloro-thieno[2,3-i7] pyrimidine dérivative and ^Æj^-arnino-d-O/ï-indolG-yljpropanoic acid as lhe appropriate amino acid dérivative, using DMA as solvent instead of DMSO, a mixture of diastereoisomers was obtained. They were separated via preparative reversed phase chromatography using 0.1 % aqueous TFA solution and acetonitrile as eluents. Example 184 was obtained as the earlier eluting diastereoisomer, HRMS calculated for C29H24N4O2S: 492.1620, found 493.1693 (M+H)

Example 185 was obtained as the later eluting diastereoisomer. HRMS calculated for C29H24N4O2S: 492.1620, found 493.1690 (M+H)

Example 186: 7V-[6-ethyl-5-(naphtlialen-l-yl)thieno[2,3-rf]pyrimidin-4-yI]-3naphthalen-l-yl-D-alanine, diastereoisomer 1

ORIGINAL

-I28and

Example 187: Af-lô-ethyl-S-Înaphthalen-l-yOthieno^^-rflpyrimidin^-ylJ-SnaphthaIen-l-yl-Z)-alanine, diastereoisomer 2

Using General Procedure ib and Préparation 4k as the appropriate 4-chloro-thieno[2,3-i/] pyrimidine dérivative and (2Æ)-2-amino-3-(l-naphthyl)propanoic acid as the appropriate amino acid dérivative, using DMA as solvent instead of DMSO, a mixture of diastereoisomers was obtained. They were separated via préparative reversed phase chromatography using 0.1 % aqueous TFA solution and acetonitrile as eluents. Example 186 was obtained as the earlier eluting diastereoisomer. HRMS calculated for C31H25N3O2S: 503.1667, found: 504.1754 (M+H)

Example 187 was obtained as the later eluting diastereoisomer. HRMS calculated for C₃iH₂5N3O₂S: 503.1667, found: 504.1758 (M+H)

Example 188: (2Zf)-biphenyI-2-yI{[6-ethyl-5-(naphthalen-l-yl)th!eno[2,3-i/]pyrimidin4-ylJamino)ethanoic acid, diastereoisomer 2

Using General Procedure 1b and Préparation 4k as the appropriate 4-chloro-thieno[2,3-<7] pyrimidine dérivative and (7î)-amino-biphenyl-2-yl-acetic acid as the appropriate amino acid dérivative, a mixture of diastereoisomers was obtained. They were separated via préparative reversed phase chromatography using 40 mM aqueous NH₄OAc solution (pH = 4, adjusted with AcOH) and acetonitrile as eluents. Examplc 188 was obtained as the later eluting diastereoisomer. HRMS calculated for C32H25N3O2S: 515.1667, found: 516.1747 (M+H)

Examplc 189: (27?)-biphenyI-3-yl{|6-ethyl-5-(naphthalen-l-yl)thieno[2,3-i7]pyrimidin4-yl]amino)ethanoic acid, diastereoisomer 1

Using General Procedure 1b and Préparation 4k as the appropriate 4-chloro-thieno[2,3-i/| pyrimidine dérivative and (7?)-amino-biphenyl-3-yl-acctic acid as the appropriate amino acid dérivative, a mixture of diastereoisomers was obtained. They were separated via préparative reversed phase chromatography using 40 mM aqueous NH₄OAc solution (pH =

ORIGINAL

- 129 -

4, adjusted with AcOH) and acetonitrile as eluents. Example 189 was obtained as the earlier eluting diastereoisomer. HRMS calculated for C32H25N3O2S: 515.1667, found: 516.1743 (M+H)

Example 190: /V-[6-ethyl-5-(naphthalen-l-yl)thieno[2,3-rf]pyrimi(lm-4-yl]-Z)-histidine

Using General Procedure la and Préparation 4k as the appropriate 4-chloro-thieno[2,3-i/] pyrimidine dérivative and (2/?)-2-amino-3-(l/7-imidazol-4-yl)propanoic acid as the appropriate amino acid dérivative, a mixture of diastereoisomers was obtained. It was purified via préparative reversed phase chromatography using 0.1 % aqueous TFA solution and acetonitrile as eluents. Example 190 was obtained as a mixture of diastereoisomers. HRMS calculated for C24H21N5O2S: 443.1416, found: 444.1462 and 444.1471 for the two diastereoisomers (M+H)

Example 191: A-|6-ethyl-5-(naplithalen-l-yI)tliieno|2,3-i/|pyriinidin-4-yl|-3-pyridin-2yI-£>-alanine

Using General Procedure ib and Préparation 4k as the appropriate 4-chloro-thieno[2,3-rZ] pyrimidine dérivative and (2/?)-2-amino-3-(2-pyridyl)propanoic acid as the appropriate amino acid dérivative, a mixture of diastereoisomers was obtained. It was purified via préparative reversed phase chromatography using 0.1 % aqueous TFA solution and acetonitrile as eluents. Examplc 191 was obtained as a mixture of diastereoisomers. HRMS calculated for C26H22N4O2S: 454.1463, found: 455.1537 and 455.1558 for the two diastereoisomers (M+H)

Examplc 192: A-|6-ethyl-5-(naphthaleri-l-yl)thieno|2,3-i/|pyrimidin-4-yl]-3-pyridin-3yI-Z)-aIanine

Using General Procedure 1b and Préparation 4k as the appropriate 4-chloro-thieno[2.3-ii^rJ pyrimidine dérivative and 3-(3-pyridyl)-£>-alanine as the appropriate amino acid dérivative, a mixture of diastereoisomers was obtained. It was purified via préparative reversed phase chromatography using 0.1 % aqueous TFA solution and acetonitrile as eluents.

ORIGINAL

- 130Example 192 was obtained as a mixture of diastereoisomers. HRMS calculated for C26H22N4O2S: 454.1445, found: 455.1545 and 455.1553 for the two diastereoisomers (M+H)

Example 193: 7V-[6-ethyl-5-(naphthalen-l-yI)thieno|2,3-r/|pyrimidin-4-yl]-3-pyridin-4yl-D-alaninc

Using General Procedure Ib and Préparation 4k as the appropriate 4-chloro-thieno[2,3-i/] pyrimidine dérivative and (27?)-2-amino-3-(4-pyridyl)propanoic acid as the appropriate amino acid dérivative, a mixture of diastereoisomers was obtained. It was purified via preparative reversed phase chromatography using 0.1 % aqueous TFA solution and acetonitrile as eluents. Example 193 was obtained as a mixture of diastereoisomers. HRMS calculated for C26H22N4O2S: 454.1440, found: 455.1540 and 455.1545 for the two diastereoisomers (M+H)

Example 194: /V-[6-ethyl-5-(naphthalen-l-yl)thieno[2,3-rf|pyrimidin-4-yl]-l-methyl-Z)histidinc

Using General Procedure Ib and Préparation 4k as the appropriate 4-chloro-thieno[2,3-<| pyrimidine derivative and (27î)-2-amino-3-(l-methylimidazol-4-yl)propanoic acid as the appropriate amino acid derivative, a mixture of diastereoisomers was obtained. It was purified via preparative reversed phase chromatography using 0.1 % aqueous TFA solution and acetonitrile as eluents. Example 194 was obtained as a mixture of diastereoisomers. HRMS calculated for C25H23N5O2S: 457.1572, found: 458.1641 and 458.1654 for the two diastereoisomers (M+H)

Example 195: l-benzyl-A^f-[6-ethyl-5-(naphthalen-l-yl)tliicno[2,3-r/]pyrimidin-4-yl]-Z)histidine

Using General Procedure Ib and Préparation 4k as the appropriate 4-chloro-thieno[2,3-t/] pyrimidine derivative and (27?)-2-amino-3-(l-benzylimidazol-4-yl)propanoic acid as the appropriate amino acid derivative, a mixture of diastereoisomers was obtained. It was

ORIGINAL

- I3l purified via preparative reversed phase chromatography using 0.1 % aqueous TFA solution and acetonitrile as eluents. Example 195 was obtained as a mixture of diastereoisomers. HRMS calculated for C31H27N5O2S: 533.1885, found: 534.1934 and 534.1934 for the two diastereoisomers (M+H)

Example 196: 7V-|6-methyl-5-(naphthalen-l-yI)thieno[2,3-rf]pyrimidin-4-yl]-Z)phenylalanine

Using General Procedure la and Préparation 41 as the appropriate 4-chloro-thieno[2,3-i7] pyrimidine dérivative and £>-phenylalanine as the appropriate amino acid dérivative, a mixture of diastereoisomers was obtained. It was purified via preparative reversed phase chromatography using 0.1 % aqueous TFA solution and acetonitrile as eluents. Example 196 was obtained as a mixture of diastereoisomers. HRMS calculated for C26H21N3O2S: 439.1354, found: 440.1421 and 440.1429 (M+H)

Example 197: 'V-j6-(hydroxymethyl)-(5Æ„)-5-(naphthalen-l-vl)thieno|2,3-i/] pyrimidin-4-yI]-Z)-plienylalanine and

Example 198: 7V-[6-(hydroxymethyl)-(5S_fl)-5-(naphthalen-l-yl)thieno[2,3-i/]pyrimidin4-yl|-Z)-phenylalanine

Using General Procedure la and Préparation 4m as the appropriate 4-chloro-thieno[2,3-i7] pyrimidine dérivative and Ώ-phenylalanine as the appropriate amino acid dérivative, a mixture of diastereoisomers was obtained. They were separated via preparative reversed phase chromatography using 0.1 % aqueous TFA solution and acetonitrile as eluents. Example 197 was obtained as the earlier eluting diastereoisomer. HRMS calculated for C26H21N3O3S: 455.1304, found: 456,1356 (M+H)

Example 198 was obtained as the later eluting diastereoisomer. HRMS calculated for C26H21N3O3S: 455.1304, found: 456.1390 (M+H)

Example 199: /V-|6-acetyl-(5S„)-5-(naphthalen-l-y[)thieno|2,3-i/]pyrimidin-4-yl]-Z)phenylalanïne

ORIGINAL

- 132 and

Example 200: N“[6-acetyl-(57î_a)-5-(naphthalen-l-yl)thieno[2,3-i/]pyriinidin-4-yl]-/)phenylalanine

Using General Procedure la and Préparation 4o as the appropriate 4-chloro-thieno[2,3-rf] pyrimidine dérivative and Z)-phenylalanine as the appropriate amino acid dérivative, a mixture of diastereoisomers was obtained. They were separated via préparative reversed phase chromatography using 40 mM aqueous NH₄OAc solution (pH = 4, adjusted with AcOH) and acetonitrile as eluents. Example 199 was obtained as the earlier eluting diastereoisomer. HRMS calculated for C27H21N3O3S: 467.1304, found: 468.1379 (M+H) Example 200 was obtained as the later eluting diastereoisomer. HRMS calculated for C27H21N3O3S: 467.1304, found: 468.1377 (M+H)

Example 201; N-|5-(naphthalen-l-yl)-6-(propan-2-yl)thieno|2,3-rf]pyrimidin-4-ylj-Z)phenylalanine, diastereoisomer 1 and

Example 202: jV-[5-(naphthalen-l-yI)-6-(propan-2-yl)thieno[2,3-rf]pyrimidin-4-yl]-Z)phenylalanine, diastereoisomer 2

Using General Procedure Ib and Préparation 4q as the appropriate 4-chloro-thieno[2,3-i/] pyrimidine dérivative and £>-phenylalanine as the appropriate amino acid dérivative, a mixture of diastereoisomers was obtained. They were separated via préparative reversed phase chromatography using 40 mM aqueous NH₄OAc solution (pH = 4, adjusted with AcOH) and acetonitrile as eluents. Example 201 was obtained as the earlier eluting diastereoisomer. HRMS calculated for C28H25N3O2S: 467.1667, found: 468.1731 (M+H) Example 202 was obtained as the later eluting diastereoisomer. HRMS calculated for C28H25N3O2S: 467.1667, found: 468.1720 (M+H)

Example 203: 7V-[6-(l-hydroxyethyl)-5-(naphthalen-l-yl)thieno[2,3-r/|pyrimidin-4-yI]D-phenylalanine, diastereoisomer 1 and

ORIGINAL

- 133Example 204: 7V-[6-(l-hydroxyethyl)-5-(naphthalen-l-yl)thieno[2,3-rf]pyrimidin-4-yl]D-phenylalanine, diastereoisomer 2

Using General Procedure la and Préparation 4nl as the appropriate 4-chloro-thieno [2,3-iZ]pyriniidine dérivative and D-phenylalanine as the appropriate amino acid dérivative, a mixture of diastereoisomers was obtained. They were separated via préparative reversed phase chromatography using O.l % aqueous TFA solution and acetonitrile as eluents. Example 203 was obtained as the earlier eluting diastereoisomer. HRMS calculated for C27H23N3O3S: 469.1460, found: 470.1511 (M+H)

Example 204 was obtained as the later eluting diastereoisomer. HRMS calculated for C27H23N3O3S: 469.1460, found: 470.1536 (M+H)

Example 205: /V-[6-(l-hydroxyethyI)-5-(naphthaIen-1-yl)thieno|2,3-i/|pyrimidin-4-yl|Z)-phenylalanine, diastereoisomer 3 and

Example 206: .'V-[6-(l-hydroxyethyl)-5-(naphthak*n-l-yl)thieno|2,3-//|pyriniidin-4-yl]Z)-phenylalanine, diastereoisomer 4

Using General Procedure la and Préparation 4n2 as the appropriate 4-chloro-thieno [2,3-i7]pyrimidine dérivative and D-phenylalanine as the appropriate amino acid dérivative, a mixture of diastereoisomers was obtained. They were separated via préparative reversed phase chromatography using 0.1 % aqueous TFA solution and acetonitrile as eluents. Example 205 was obtained as the earlier eluting diastereoisomer. HRMS calculated for C27H23N3O3S: 469.1460, found: 470.1539 (M+H)

Example 206 was obtained as the later eluting diastereoisomer. HRMS calculated for C27H33N3O3S: 469.1460, found: 470.1534 (M+H)

Example 207: /V-[6-(difluoromethyl)-5-(naphthaIcn-l-yl)thieno[2,3-<7]pyrimidin-4-yl|2>-phenylalanine, diastereoisomer 1 and

Example 208: 7V-[6-(difiuoromethyl)-5-(naphthalen-l-yl)thieno[2,3-rf|pyriniidin-4-yI]D-phenylalanine, diastereoisomer 2

ORIGINAL

- 134Using General Procedure la and Préparation 4r as the appropriate 4-chloro-thieno[2,3-i/] pyrimidine derivative and D-phenylalanine as the appropriate amino acid derivative, a mixture of diastereoisomers was obtained. They were separated via préparative reversed phase chromatography using 40 mM aqueous NHjOAc solution (pH = 4, adjusted with AcOH) and acetonitrile as eluents. Example 207 was obtained as the earlier eluting diastereoisomer. HRMS calculated for C26H19F2N3O2S; 475.1166, found: 476.1242 (M+H) Example 208 was obtained as the later eluting diastereoisomer. HRMS calculated for C26H19F2N3O2S: 475.1166, found: 476.1244 (M+H)

Example 209: A/-|6-(2-hydroxypropan-2-yl)-(5Æ_ü)-5-(naphthaIen-l-yl)thieno[2,3-rf] pyrimidin-4-yl]-Z>-phcnyIalanine and

Example 210: 2V-[6-(2-hydroxypropan-2-yl)-(5.S’J-5-(naphthalen-l-Yl)thieno|2,3-//| pyrimidin-4-yl]-Z>-phenylaianine

Using General Procedure la and Préparation 4p as the appropriate 4-chloro-thieno[2,3-iZ] pyrimidine derivative and D-phenylalanine as the appropriate amino acid derivative. a mixture of diastereoisomers was obtained. They were separated via préparative reversed phase chromatography using 0.1 % aqueous TFA solution and acetonitrile as eluents. Example 209 was obtained as the earlier eluting diastereoisomer. HRMS calculated for C28H25N3O3S: 483.1617, found: 484.1689 (M+H)

Example 210 was obtained as the later eluting diastereoisomer. HRMS calculated For C28H25N3O3S: 483.1617, found: 484.1704 (M+H)

Example 211: A^r-|6-iodo-5-(naphthalen-l-yl)thieno|2,3-i/]pyrimidin-4-yl]-Z>phenylalanine

Using General Procedure la and Préparation 4s as the appropriate 4-chloro-thieno[2,3-<7] pyrimidine derivative and D-phenylalanine as the appropriate amino acid derivative, using DMA as solvent instead of DMSO, a mixture of diastereoisomers was obtained. It was purified via préparative reversed phase chromatography using 40 mM aqueous NH₄OAc

ORIGINAL • I35 solution (pH = 4, adjusted with AcOH) and acetonitrile as eluents. Example 211 was obtained as a mixture of diastereoisomers. HRMS calculated for CisHigINjOiS: 551.0164, found: 552.0258 (M+H)

Example 212: /V-[(5/f₀)-5-(3-chloro-2-methylphenyl)-6-cthenyl-thieno|2,3-(/I pyrimidin-4-yI|-D-phenylaIanine and

Example 213: /V-[(5S'_fl)-5-(3-chloro-2-niethyIplienyl)-6-ethenyl-thieno[2,3-J]pyrimidin4-yl]-£>-phenylalanine

Using General Procedure Ile and Préparation 5a as the the appropriate 6-iodo-thieno [2,3-</]pyrimidine and vinyiboronic acid pinacol ester as the appropriate boronic acid dérivative, a mixture of diastereoisomers was obtained. They were separated via preparative reversed phase chromatography using 0.1 % aqueous TFA solution and acetonitrile as eluents. Example 212 was obtained as the earlier eluting diastereoisomer. HRMS calculated for C24FI20CIN3O2S: 449.0965, found: 450.1038 (M+H)

Example 213 was obtained as the later eluting diastereoisomer. HRMS calculated for C24H20CIN3O2S: 449.0965, found: 450.1050 (M+H)

Example 214; 7V-I(5Æ_ü)-5-(3-chloro-2-methylphcnyl)-6-(prop-l-cn-2-yl)thieno[2,3-i/| pyrimidin-4-yl]-Z)-phenylaIanine and

Example 215: .'V-|(5.S’„)-5-(3-chloro-2-nieth>lphenyl)-6-(prop-l -en-2-yl)thieno|2,3-<7] pyrimidin-4-yl|-Z>-phenylaIanine

Using General Procedure Ile and Préparation 5a as the the appropriate 6-iodo-thieno [2,3-i/]pyrimidine and 2-isopropenyl-4,4,5,5-tetramethyl-l,3,2-dioxaborolane as the appropriate boronic acid dérivative, a mixture of diastereoisomers was obtained. They were separated via preparative reversed phase chromatography using 0.1 % aqueous TFA solution and acetonitrile as eluents. Example 214 was obtained as the earlier eluting diastereoisomer. HRMS calculated for C25H22CIN3O2S; 463.1121, found: 464.1178 (M+H)

ORIGINAL

- 136 Example 215 was obtained as the later eluting diastereoisomer. HRMS calculated for C25H22CIN3O2S: 463.1121, found: 464.1179 (M+H)

Example 216: A-[(57f_fl)-5-(3-chIoro-2-methylphenyl)-6-cyclopropyl-thieno|2,3-i/| pyrimidin“4-yl]-Z>-phenylalanine and

Example 217: jV-[(55_fl)-5-(3-chIoro-2-methylphenyl)-6-cyclopropyl-thieno[2,3-i/| pyrimidin-4-yl]-D-phenylalanine

Using General Procedure lia and Préparation 5a as the the appropriate 6-iodo-thieno [2,3-t/]pyrimidine and cyclopropylboronic acid as the appropriate boronic acid dérivative, 10 and Bu^NOH instead of K2CO3, a mixture of diastereoisomers was obtained. They were separated via préparative reversed phase chromatography using 0.1 % aqueous TFA solution and acetonitrile as eluents. Example 216 was obtained as the earlier eluting diastereoisomer. HRMS calculated for C25H22CIN3O2S: 463.1121, found: 464.1177 (M+H) Example 217 was obtained as the later eluting diastereoisomer. HRMS calculated for 15 C25H22CIN3O2S: 463.1121, found: 464.1182 (M+H)

Example 218: /V-[(55'_i()-5-(3-chloro-2-niethYlphcnyl)-6-(propan-2-yl)thieno[2,3-J] pyrimidin-4-yl|-L-phenylalanine and

Example 219: 7V-[(5Jf_a)-5-(3-chloro-2-methylphenyl)-6-(propan-2-yl)thieno[2,3-f/| 20 pyrimidin-4-yl|-L-phenylalanine

Using General Procedure 1b and Préparation 4u as the the appropriate 4-chlorothieno[2,3-t/]pyrimidine dérivative and £-phenylalanine as the appropriate amino acid dérivative, a mixture of diastereoisomers was obtained. They were separated via préparative reversed phase chromatography using 0.1 % aqueous TFA solution and 25 acetonitrile as eluents. Examplc 218 was obtained as the earlier eluting diastereoisomer.

HRMS calculated for C25H24CIN3O2S: 465.1278, found: 466.1371 (M+H)

Example 219 was obtained as the later eluting diastereoisomer. HRMS calculated for C25H24CIN3O2S: 465.1278, found: 466.1361 (M+H)

ORIGINAL

- 137Example 220: /V-[(5/G)'5-(3-chIoro-2-methyIphenyl)-6-(propan-2-yl)thieno|2,3-rfJ pyrimidin-4-yl]-Z)-phenylalanine and

Example 221 : AH(5Sj-5-(3-chloro-2-methylphenyl)-6-(prop;iii-2-yl)thieno|2.3-<7| pyrimidin-4-yl]-Z)-phcnylalanine

Using General Procedure Ib and Préparation 4u as the the appropriate 4-chlorothieno[2,3-i/]pyrimidine dérivative and £)-phenylalanine as the appropriate amino acid derivative, a mixture of diastereoisomers was obtained. They were separated via préparative reversed phase chromatography using 0.1 % aqueous TFA solution and acetonitrile as eluents. Example 220 was obtained as the earlier eluting diastereoisomer. HRMS calculated for C25H24ON3O2S: 465.1278, found: 466.1348 (M+H)

Example 221 was obtained as the later eluting diastereoisomer. HRMS calculated for C25H24CIN3O2S: 465.1278, found: 466.1350 (M+H)

Example 222: A'-|(5/?_Îj-5-(3-chloi-o-2-ni€thylphenyl)-6-(pr()pan-2-yl)thieno|2,3-r/] pyrimidin-4-yI]-2-methoxy-Z>-phenylalanine and

Example 223: N-((55_e)-5-(3*chloro-2-methy]phenyl)-6-(propan-2-yl)thieno|2,3-4/] pyrimidin-4-yl|-2-methoxy-Z)-plienylalanine

Using General Procedure Ib and Préparation 4u as the the appropriate 4-chlorothieno[2,3-i/]pyrimidine derivative and (2Æ)-2-amino-3-(2-methoxyphenyl)piOpanoic acid as the appropriate amino acid derivative, a mixture of diastereoisomers was obtained. They were separated via préparative reversed phase chromatography using 0.1 % aqueous TFA solution and acetonitrile as eluents. Example 222 was obtained as the earlier eluting diastereoisomer. HRMS calculated for C26H26CIN3O3S: 495.1383, found: 496.1460 (M+H) Example 223 was obtained as the later eluting diastereoisomer. FIRMS calculated for C26H26CIN3O3S: 495.1383, found: 496.1454 (M+H)

ORIGINAL

- 138Example 224: A/-[(5ff_(/)-5-(3-chloro-17f-indol-4-yl)-6-ethyl-thieno|2,3-rf]pyrimidin-4yl]-Z)-phenylalanine and

Example 225: (V-[(5_<S'_t,)-5-(3-chloro-l//-indol-4-yl)-6-ethyl-thieno[2,3-r/]pynmÎdin-4yl|-Z)-phenylalanine

522 mg Préparation 7h (I mmol), 164 mg NCS (1.2 mmol), 15 mL CCl₄ and 10 mL THF were stirred at r.t. under N? atmosphère tor 2 hours. Then the mixture was poured into icy water and extracted with DCM. The combined organic phases were dried over Na₂SO₄, filtered and the filtrate was concentrated under reduced pressure. The formed diastereoisomers were separated via preparative reversed phase chromatography using 40 mM aqueous NH₄OAc solution (pH - 4, adjusted with AcOH) and acetonitrile as eluents. Example 224 was obtained as the later eiuting diastereoisomer.HRMS calculated for C25H2iClN₄O2S: 476.1074, found: 477.1133 (M+H)

Example 225 was obtained as the earlier eiuting diastereoisomer. HRMS calculated for C₂₅H₂iC1N₄O₂S: 476.1074, found: 477.1137 (M+H)

Example 226: jV-[(5S'_a)-5-(3-bromo-17f-indol-4-yl)-6-ethyIthieno[2,3-i/|pyrimidin-4y[|-Z>-phenylaIanine and

Example 227: A-|(5/?_(J)-5-(3-bromo-l//-indol-4-yI)-6-etliylthicno[2,3-i'/]pyrimidin-4yl|-Z)-phenylalanine

522 mg Préparation 7h (1 mmol), 216 mg NBS (1.2 mmol), 15 mL CC1₄ and 5 mL THF were stirred at r.t. under N2 atmosphère for 2 hours. Then the mixture was poured into icy water and extracted with DCM. The combined organic phases were dried over Na2SO₄, filtered and the filtrate was concentrated under reduced pressure. The formed diastereoisomers were separated via preparative reversed phase chromatography using 40 mM aqueous NH₄OAc solution (pH = 4, adjusted with AcOH) and acetonitrile as eluents. Example 226 was obtained as the earlier eiuting diastereoisomer. HRMS calculated for C₂₅H₂₁BrN₄O₂S: 520.0569, found: 521.0653 (M+H)

ORIGINAL

- I39Example 227 was obtained as the later eluting diastereoisomer. HRMS calculated for C₂₅H_2lBrN₄O₂S: 520.0569, found: 521.0629 (M+H)

Example 228: /V-[6-ethyl-(5S„)-5-(3-io(10“l/7-intlol-4-yl)thieno[2,3-i/]pyrinüdin-4-yl]£>-phenylalanine and

Example 229: 7V-|6-ethyl-{5/f_a)-5-(3-iodo-l//-indol-4-yl)thieno|2,3-i/|pyrimidin-4-yl]£>-phenylalanine

522 mg Préparation 7h (1 mmol), 196 mg KOH (3.5 mmol), 15 mL DMF and 267 mg iodine (1.05 mmol) were stirred at r.t. under N₂ atmosphère for 18 hours. Then the mixture 10 was poured into icy water and saturated Na₂S₂0j solution was added. The mixture was extracted with DCM, the combined organic phases were dried over Na₂SO₄, filtered and the filtrate was concentrated under reduced pressure. The formed diastereoisomers were separated via préparative reversed phase chromatography using 40 mM aqueous NFLOAc solution (pH = 4, adjusted with AcOH) and acetonitrile as eluents. Example 228 was 15 obtained as the earlier eluting diastereoisomer. HRMS calculated for C?5H₂irN₄O₂S: 568.043, found: 569.0498 (M+H)

Example 229 was obtained as the later eluting diastereoisomer. HRMS calculated for C₂₅H₂|IN₄O₂S: 568.043, found: 569.0502 (M+H)

Example 230: 7V-((5S_a)-5-{3-chloro-l-[2-(dimethylamino)etliyl]-l//-indoI-4-yl}-620 ethyl-tlneno[2,3-i/]pyrimiilin-4-yl)-Z)-phenylalanine and

Example 231: /V-((5/i_a)-5-{3-chloro-I-[2-(dimethylamino)ethy[|-l//-indol-4-yi}-6ethyl-thieno|2,3-rf| pyrimidin-4-yI)-Z>-phenyIalaninc

Using General Procedure VIII and Préparation 7i as the appropriate indole dérivative and 25 2-(AUV-dirnethyiamino)ethanol as the appropriate alcohol, Example 230 was obtained as the earlier eluting diastereoisomer. HRMS calculated for C₂9H3₀ClN₅O₂S: 547.1809, found: 548.1902 (M+H)

ORIGINAL

- I40Example 231 was obtained as the later eluting dîastereoisomer. HRMS calculated for C29H30CIN5O2S: 547.1809, found: 548.1889 (M+H)

Example 232: A-((5/?_ÎÎ)-5-{3-clilor<>-l-|2-(pyrrolidin-Lyl)ethyl|-l//-indol-4-yl}-6ethyl-thieno|2,3-r/|pyrimidin-4-yl)-£)-phenylalanÎne and

Example 233: /V-((55_a)-5-{3-chloro-l-[2-(pyrrolidin-l-yl)ethyl]-l£f-indoI-4-yl}-6-ethylthieno[2,3-J|pyrimidin-4-yI)-Z>-phenylaIanine

Using General Procedure VIII and Préparation 7i as the appropriate indole dérivative and 2-pyrrolidin-l-ylethanol as the appropriate alcohol. Example 232 was obtained as the later eluting dîastereoisomer. HRMS calculated for CjiHjiClNsCbS: 573.1965, found: 574.2059 (M+H)

Example 233 was obtained as the earlier eluting dîastereoisomer. HRMS calculated for C31H32CIN5O2S: 573.1965, found: 574.2060 (M+H)

Example 234: A^r-((5/f_a)-5-{3-chloro-l-J2-(piperidin-l-yl)ctbylj-l/7-indol-4-vl|-6-ethylthieno[2,3-rf|pyrimidin-4-vl)-Z)-phenyIalanine and

Example 235: W-((55_rt)-5-{3-chloro-l-[2-(piperidin-l-yl)cthyl]-lÆ-indol-4-yl}-6-ethylthieno[2,3-i/|pyrimidin-4-yl)-Z)-phenylalanine

Using General Procedure VIII and Préparation 7i as the appropriate indole dérivative and 2-(l-piperidyl)ethanol as the appropriate alcohol, a mixture of diastereoisomers was obtained. They were separated via préparative reversed phase chromatography using 0.1 % aqueous TFA solution and acetonitrile as eluents. Example 234 was obtained as the earlier eluting dîastereoisomer. HRMS calculated for C32H34CIN5O2S: 587.2122, found: 588.2201 (M+H)

Example 235 was obtained as the later eluting dîastereoisomer. HRMS calculated for C32H34CIN5O2S: 587.2122, found: 588.2199 (M+H)

ORIGINAL

- I4l Example 236: ^-((SÆJ-Â-ÎS-chloro-l-^-tmorpholin^-yOethyll-lH-indol^-ylJ-ôethyl-thieno[2,3-rf]pyrimidin-4-yI)-I>-phenylalanine and

Example 237: A^r-((55'_o)-5-{3-chloro-I-|2-(morpholin-4-yI)ethyiJ-lÂ/-indol-4-yl}-6ethyl-thieno|2,3-i/]pyrimidin-4-yl)-2)-phenylalanine

Using General Procedure VIII and Préparation 7i as the appropriate indole derivative and 2-morphoIinoethanol as the appropriate alcohol, Example 236 was obtained as the later eluting diastereoisomer. HRMS calculated for C31H32CIN5O3S: 589.1914, found: 590.1998 (M+H)

Example 237 was obtained as the earlier eluting diastereoisomer. HRMS calculated for C31H32CIN5O3S: 589.1914, found: 590.1994 (M+H).

Example 238: 7V-((55_fl)-5-{3-chloro-l-[2-(4-methyIpiperazin-l-yl)ethyl|-l/7-indol-4yI}-6-ethyl-thieno[2,3-i/]pyrimidin-4-yl)-D-phenyIaIanine and

Example 239: .'V-((5/?„)-5-{3-chloro-l-[2-(4-metIiylpiperazin-l-yl)ethyl|-l//-indoI-4yl}-6-ethyl-thieno[2,3-rf] pyrimidin-4-yl)-£>-phenylalanine

Using General Procedure VIII and Préparation 7i as the appropriate indole derivative and 2-(4-methylpiperazin-l-yl)ethanol as the appropriate alcohol. Exampte 238 was obtained as the earlier eluting diastereoisomer. HRMS calculated for C32H35CIN6O2S: 602.2231, found: 603.2312 (M+H)

Example 239 was obtained as the later eluting diastereoisomer. HRMS calculated for C32H35CIN6O2S: 602.2231, found: 603.2311 (M+H)

Example 240: ÎV-((5>S_fl)-5-{3-chIoro-l-[3-(4-inetliylpiperazin-l-yl)propyI]-l//-indol-4yl}-6-ethyl-thieno|2,3-i/|pyrimidin-4-yl)-Z)-phenylalanine and

Exaniple 241: jV-((57f_a)-5-{3-chloro-l-[3-(4-methyIpiperazin-l-yl)propylJ-l//-Îndol-4yl}-6-ethyl-thieno[2,3-rf]pyrimidin-4-yI)-/)-phenyIalanine

ORIGINAL

- 142Using General Procedure VIII and Préparation 7i as the appropriate indole dérivative and

3-(4-methylpiperazin-l-yl)propan-l-ol as the appropriate alcohol, Example 240 was obtained as the earlier eluting diastereoisomer. HRMS calculated for C33H37CIN6O2S: 616.2387, found: 617.2466 (M+H)

Example 241 was obtained as the later eluting diastereoisomer. HRMS calculated for C₃3H37C1N₆O₂S: 616.2387, found: 617.2473 (M+H)

Example 242: 3-cyclohexyl-jV-|6-ethyl-5-(lH-indol-4-yl)thieno[2,3-//]pyrimidin-4-yl|Z)-alanine, diastereoisomer 1 and

Example 243: 3-cyclohexyl-jV-|6-ethyl-5-(lH-indol-4-yl)thieno[2,3-i/|pyrimidin-4-yl|Z)-alanine, diastereoisomer 2

Using General Procedure la and Préparation 4v as the appropriate 4-chloro-thieno[2,3-t/| pyrimidine dérivative and (2Æ)-2-amino-3-cyclohexyl-propanoic acid as the appropriate amino acid dérivative, a mixture of diastereoisomers was obtained. They were separated via préparative reversed phase chromatography using 0.1 % aqueous TFA solution and acetonitrile as eluents. Example 242 was obtained as the earlier eluting diastereoisomer. HRMS calculated for C25H28N4O2S: 448.1933, found: 449.1994 (M+H)

Example 243 was obtained as the later eluting diastereoisomer. HRMS calculated for C25H₂₈N4O₂S: 448.1933, found: 449.2006 (M+H)

Example 244: /V-[(55'_n)-5-(3-chloro-I/7-indol-4-yI)-6-ethyI-thieno[2,3“i/|pyrimidin-4yl]-3-pyridin-2-yl-Z)-alanine and

Example 245: 7V-|(5Jî₀)-5-(3-chIoro-l//-indol-4-yl)-6-ethyl-thieno|2,3-rf|pyrimidin-4yI]-3-pyridin-2-yl-Z)-alanine

Using General Procedure la and Préparation 7j as the appropriate 4“Chloro-thieno[2,3-<7] pyrimidine dérivative and (2Æ)-2-amino-3-(2-pyridyl)propanoic acid as the appropriate amino acid dérivative, a mixture of diastereoisomers was obtained. They were separated via préparative reversed phase chromatography using 40 mM aqueous NH4OAC solution

ORIGINAL

- I43 (pH - 4, adjusted with AcOH) and acetonitrile as eluents. Example 244 was obtained as the earlier eluting diastereoisomer. HRMS calculated for C24H20CIN5O2S: 477.1026, found: 478.1087 (M+H)

Example 245 was obtained as the later eluting diastereoisomer. HRMS calculated for C24H20CIN5O2S: 477.1026, found: 478.1089 (M+H)

Example 246: 7V-((5S_a)-5-{3-chloro-l-[2-(morpholin-4-yl)ethyl]-17/-indol-4-yl}-6ethyl-thÎeno[2,3-</]pyrimidm-4-yl)-3-pyridin-2-yLD-alaninc

Step A: methyl (2R)-2-[[(5S-J-5-('3-chloro-l\ï-indol-4-yl)-6-ethyl-thieno[2,3-d]pyrimidin-

4-yl]amino]-3-(2-pyridyl)propanoate

0.13 g Example 244 (0.27 mmol) was dissolved in 13 mL MeOH, then 0.3 mL cc. H₂SO₄was added and the mixture was stirred at r.t. until no further conversion was observed. Then it was concentrated under reduced pressure and saturated aqueous NaHCOj solution was added and the mixture was agitated. The formed precipitate was collected by filtration to obtain methyl (27/)-2-(((5.5^)-5-(3-01101^0-1 7/-indol-4-yl)-6-ethyl-thieno[2,3-i7] pyrimidin-4-yl]amino]-3-(2-pyridyl)propanoate.

Step B: Example 246

Using General procedure VIII and methyl (2/?)-2-[[(5S_a)-5-(3-chloro-17/-indol-4-yl)-6ethyl-thieno[2,3-iZ]pyrimidin-4-yl]amino]-3-(2-pyridyl)propanoate as the appropriate indole dérivative and 2-morpholinoethanol as the appropriate alcohol, Example 246 was obtained. HRMS calculated for CsoHjiCINûOsS: 590.1867, found: 591.1938 (M+H)

Example 247: 7V-((5Æ_fi)-5-{3-cliloro-l-|2-(morpholin-4-yl)ethyl|-l 7/-indol-4-yl|-6cthyl-thieno|2,3-i/|pyrimidin-4-yl)-3-pyridin-2-yl-Z)-aIanine

Step A: methyl (2R)-2-[[(5R₃)-5-(3-chloro-l\)-indol-4-yl)-6-ethyl-thieno[2,3-<3]pyrimidin4-yl]amino]-3-(2-pyridyl)propanoate

0.157 g Example 245 (0.33 mmol) was dissolved in 15 mL MeOH, then 0.3 mL cc. H2SO4 was added and the mixture was stirred at r.t. until no further conversion was observed. Then it was concentrated under reduced pressure and saturated aqueous NaHCOj solution

ORIGINAL

- 144was added and the mixture was agitated. The formed precipitate was collected by filtration to obtain methyl (2/?)-2-[[(5Æ₀)-5-(3-chloro-l//-indol-4-yl)-6-ethyl-thieno[2,3-if| pyrimidin-4-yl]amino]-3-(2-pyridyl)propanoate.

Step B: Example 247

Using General procedure VIII and methyl (2Æ)-2-[[(5Æ_a)-5-(3-chIoro-1 /7-indol-4-yl)-6ethyl-thieno[2,3-i/]pyrimidin-4-yl]amino]-3-(2-pyridyl)propanoate as the appropriate amine and 2-morpholinoethanol as the appropriate alcohol Example 247 was obtained. HRMS calculated for C₃₀H₃iClN₆O₃S: 590.1867, found: 591.1918 (M+H)

Example 248: A'-((5₁S'„)-5-{3-chloro-l-|2-(4-mcthylpiperazin-l-yl)ethyl|-l//-indol-410 yl}-6-ethyl-thieno[2,3-rf|pyrimidin-4-yl)-3-pyridin-2-yl-Z)-alanine

Step A: methyl (2\i)-2-[[(53^)-5-(3-chloro-lY{-indol-4-yl)-6-ethyl-thieno[2,3-à]pyrimidin4-yl] amino]-3-(2-pyridyl)propanoate

0.13 g Example 244 (0.27 mmol) was dissolved in 13 mL MeOH, then 0.3 mL cc. H₂SO₄was added and the mixture was stirred at r.t. until no further conversion was observed.

Then it was concentrated under reduced pressure and saturated aqueous NaHCO₃ solution ^was added and the mixture was agitated. The formed precipitate was collected by filtration to obtain methyl (2tf)-2-[[(5Sj-5-(3-chloro-l//-indol-4-yl)-6-ethyl-thieno[2,3-i/| pyrimidin-4-yl]amino]-3-(2-pyridyl)propanoate.

Step B: Example 248

Using General procedure VIII and methyl (2/f)-2-[[(5S_o)-5-(3-chloro-lZ/-indol-4-yl)-6ethyl-thieno[2,3-i/]pyrimidin-4-yl]amino]“3-(2-pyridyl)propanoate as the appropriate indole dérivative and 2-(4-methylpiperazin-l-yi)ethanol as the appropriate alcohol Example 248 was obtained. HRMS calculated for C₃₁H₃₄C1N₇O₂S: 603.2183, found: 302.6172 (M+2H)

Example 249: jV-((5Æ_fl)-5-{3-chloro-l-[2-(4-methylpiperazin-l-yl)ethyl|-l//-indol-4yl}-6-ethyl-thieno|2,3-rf|pyrimidin-4-yl)-3-pyiidin-2-yl-Z)-aIanine

ORIGINAL

- I45Slep A: methyl (2B^)-2-[[(5^J-5-(3-chloro-l\3-indol-4-yl)-6-ethyl-thieno[2,3-d]pyrimidin4-yl] amino]~3-(2-pyridyl)propanoate

0.157 g Example 245 (0.33 mmol) was dissolved in 15 mL MeOH, then 0.3 mL cc. H₂SO₄was added and the mixture was stirred at r.t. until no further conversion was observed. Then it was concentrated under reduced pressure and saturated aqueous NaHCO₃ solution was added and the mixture was agitated. The formed precipitate was collected by filtration to obtain methyl (2Æ)-2-[[(5Æ„)-5-(3-chloro-l//-indol-4-yl)-6-ethyl-thieno[2,3-<7] pyrimidin-4-yl]amino]-3-(2-pyridyl)propanoate.

Step B: Example 249

Using General procedure VIII and methyl (2/?)-2-[[(57?J-5-(3-chloro-l //-indol-4-y[)-6ethylthieno[2,3-i/]pyrimidin-4-yl]amino]-3-(2-pyridyl)propanoate as the appropriate indole dérivative and 2-(4-methylpiperazin-l-yl)ethanol as the appropriate alcohol Example 249 was obtained. HRMS calculated for C^lLuCIN^LS: 603.2183, found: 302.6164 (M+2H)

Example 250: jV-[6-(difluoromethyl)-(5S_i,)-5-(naphtlialen-l-yI)thieno[2,3-rf|pyi’imidin4-yl]-3-pyridin-2-yl-Z)-alanine and

Example 251: 7V-j6-(difluoromethyl)-(5./f„)-5-(naphthalen-l-yl)thieno[2,3-i7]pyrimidin4-yl|-3-pyridin-2-yl-2)-alanine

Using General Procedure la and Préparation 4r as the appropriate 4-chioro-lhieno[2,3-J| pyrimidine dérivative and (2Æ)-2-amino-3-(2-pyridyl)propanoic acid as the appropriate amino acid dérivative, a mixture of diastereoisomers was obtained. They were separaled via préparative reversed phase chromatography using 40 mM aqueous NH₄OAc solution (pH = 4, adjusted with AcOH) and acetonitrile as eluents. Example 250 was obtained as the earlier eluting diastereoisomer. HRMS calculated for C₂₅H]_gF₂N₄O2S: 476.1119, found: 477.1195 (M+H)

Example 251 was obtained as the later eluting diastereoisomer. HRMS calculated for C₂₅H_lgF₂N₄O₂S: 476.1119, found: 477.1182 (M+H)

ORIGINAL

- 146Example 252: AH(5S_fl)-5-(naphthalen-l-yl)-6-propylthieno|2,3-i/]pyrimidin-4-yl]-7>phenylalanine

266 mg Example 108 (0.57 mmol) was dissolved in 10 mL MeOH and 2 mL AcOH, then 61 mg 10% Pd/C was added. The mixture was stirred under H₂ atmosphère at 40°C for 2 hours. It was filtered through Celite and the fîltrate was concentrated under reduced pressure. The crude product was purified via préparative reversed phase chromatography using 40 mM aqueous NH4OAC solution (pH = 4, adjusted with AcOH) and acetonitrile as eluents to obtain Example 252. HRMS calculated for C28H25N3O2S: 467.1667, found: 468.1746 (M+H)

Example 253: /V-|(57i_1I)-5-(naphthalen-l-yI)-6-propylthieno[2,3-i/]pyrimidin-4-yl]-Z)phenylalanine

266 mg Example 109 (0.57 mmol) was dissolved in 10 mL MeOH and 2 mL AcOH, then 61 mg 10 % Pd/C was added. The mixture was stirred under H₂ atmosphère at 40 °C for 2 hours. It was filtered through Celite and the fîltrate was concentrated under reduced pressure. The crude product was purified via préparative reversed phase chromatography using 40 mM aqueous NH4OAC solution (pH = 4. adjusted with AcOH) and acetonitrile as eluents to obtain Example 253. HRMS calculated for C+sHiîN^OtS: 467.1667, found: 468.1736 (M+H)

Examplc 254: methyl A-|6-ethyl-(5_lS’_i/)-5-(naphtlialeii-l-yl)thicno[2,3-r/|pyrirnidin-4yl]-£>-plienyIaIaninate

102 mg Example 101 (0.225 mmol) was dissolved in 2 mL MeOH and the mixture was cooled to 0°C under N₂ atmoshphere. Then 135 pL diazomelhyl(trimethyl)silane solution (2M in Et₂O) was added and the mixture was aliowed to warm up to r.t. Then the mixture was concentrated in vacuo and purified via flash chromatography using heptane and EtOAc as eluents to obtain Examplc 254. HRMS calculated for C₂sH₂5N3O₂S: 467.1667, found: 468.1746 (M+H)

ORIGINAL

- 147Example 255: methyl /V-[6-ethy[-(5/G)-5-(naphtha!en-l-yl)thieno|2,3-rf|pyrimitiin-4j I|-£>-phenylalaninate

102 mg Example 100 (0.225 mmol) was dissolved in 2 mL MeOH and the mixture was cooled to 0 °C under N₂ atmoshphere. Then 135 pL diazomethyl(trimethyl)silane solution (2M in Et₂O) was added and the mixture was allowed to warm up to r.t. The mixture was concentrated in vacuo and purified via flash chromatography using heptane and EtOAc as eluents to obtain Example 255. HRMS calculated for C₂₈H₂5N3O₂S: 467.1667, found: 468.1737 (M+H)

Example 256: ethyl A^r-[(5S_i/)-5-{3-chloro-2-nicthyl-4-|2-(4-methylpipcrazin-l-yl) ethoxy]phenyI}-6-(4-fluorophenyl)thieno[2,3-rf]pyrinHdin-4-yl]-2-{|2-(2-methoxy phenyl)pyrimidin-4-yI|methoxy}-Z)-phenylalaninate

Example 7 was dissolved in HCl solution (20 mL/mmol, 1.25M in EtOH) and the mixture was stirred at r.t, overnight. Then the mixture was neutralized with NaHCO₂ solution and it was extracted with DCM. The organic phase was dried over Na₂SO₄, filtered and the fîltrate was concentrated under reduced pressure. The crude product was purified via preparative reversed phase chromatography using 25 mM aqueous NH4HCO3 solution and acetonitrile as eluents to give Example 256. HRMS calculated for C49H49CIFN7O5S: 901.3188, found: 902.3225 (M+H)

Examplc 257: ethyl 2-|(l-/ÉT/-butyl-l/7-pyrazoI-5-yl)methoxy|-_JV-|(5S„)-5-{3-chloro-2methyl-4-|2-(4-methylpiperazin-l-yl)ethoxy|phenyl}-6-(prop-l-yn-l-yl)thieno[2,3-rf| pyrimidin-4-ylj-D-phenylalaninate

Example 40 was dissolved in HCl solution (20 mL/mmol, 1.25M in EtOH) and the mixture was stirred at r.t. overnight. Then the mixture was neutralized with NaHCÛ3 solution and it was extracted with DCM. The organic phase was dried over Na₂SO₄, filtered and the fîltrate was concentrated under reduced pressure. The crude product was purified via preparative reversed phase chromatography using 25 mM aqueous NH4HCO3

ORIGINAL

- 148solution and acetonitrile as eluents to give Example 257. HRMS calculated for C42H₅₀ClN7O4S: 783.3334, found: 392.6744 (M+2H)

Example 258: ethyl /V-|(5.S'_1J)-5-{3-chloro-2-nicthyI-4-[2-(4-methylpiperazin-l-yI) ethoxy]phenyl}-6-(prop-l-yn-l-yl)thieno[2,3-i/|pynmidin-4-yl]-2-{|2-(2-methoxy phenyl)pyrimidin-4-yl|methoxy}-Z)-phenylalaninate

Example 45 was dissolved in HCl solution (20 mL/mmol, 1.25M in EtOH) and the mixture was stirred at r.t. ovemight. Then the mixture was neutralized with NaHCOs solution and it was extracted with DCM. The organic phase was dried over Na2SO₄, filtered and the filtrate w'as concentrated under reduced pressure. The crude product was purified via preparative reversed phase chromatography using 25 mM aqueous NH4HCO3 solution and acetonitrile as eluents to give Example 258. HRMS calculated for C46H48CIN7O5S: 845.3126, found: 423.6650 (M+H)

Example 259: ethyl jV-|(55_fl)-5-{3-chloro-4-[2-(dimethylaniino)ethoxyJ-2-methyl phenyI}-6-(prop-l-yn-l-yl)thieno[2,3-rf]pyrimidin-4-yl]-2-{|2-(2-methoxyphenyl) pyrimidin-4-yl]methoxy}-Z>-plienylalanmate

Example 49 was dissolved in HCl solution (20 mL/mmol, 1.25M in EtOH) and the mixture was stirred at r.t. ovemight. Then the mixture was neutralized with NaHCO₃solution and it was extracted with DCM. The organic phase was dried over Na2SÛ4. filtered and the filtrate was concentrated under reduced pressure. The crude product was purified via preparative reversed phase chromatography using 25 mM aqueous NH4HCO3 solution and acetonitrile as eluents to give Example 259. HRMS calculated fbr C₄3H₄3C1N₆O₅S: 790.2704, found: 396.1425 (M+2H)

Example 260: ethyl /V-|(5S_e)-5-{3-chloro-4-[2-(dimethylanuno)ethoxy[-2-methyl phenyl}-6'(4-fluorophenyl)thieno|2,3-r/|pyrimidin-4-yl]-2-({2-J2-(2-methoxyethoxy) phenyl]pyrimidin-4-yI}methoxy)-Z)-phenylalaninate

ORIGINAL

- I49Example 51 was dissolved in HCl solution (20 mL/mmol, 1.25M in EtOH) and the mixture was stirred at 60 °C until no further conversion was observed. Then the mixture was neutralized with NaHCOa solution and it was extracted with DCM. The organic phase was dried over Na2SO₄, filtered and the filtrate was concentrated under reduced pressure.

The crude product was purified via preparative reversed phase chromatography using 25 mM aqueous NH4HCO3 solution and acetonitrile as eluents to obtain Exaniple 260. HRMS calculated for C₄₈H_4SC1FN₆O₆S: 890.3029, found: 891.3105 (M+H)

Example 261: (5-methyI-2-oxo-l,3-dioxol-4-yl)methyl 7ν-[(55_σ)-5-{3-€ΗΙθΓο-4-|2(dimetliylamino)ethoxy]-2-methylphenyi}-6-(4-fluorophenyl)thieno[2,3-</]pyriniidin10 4-yl|-2-({2-|2-(2-iîicthoxyethoxy)phenyl]pyrimidin-4-yl}metlioxy)-Z>-plienylalaninate eq. Example 51 and 1.1 eq. 4-(chloromethyl)-5-methyl-l,3-dioxol-2-one were dissolved in DMF (10 mL/mmol), then 2 eq. Nal and 2 eq. CS2CO3 were added and the mixture was stirred until no further conversion was observed. Then the mixture was directly injected and purified via preparative reversed phase chromatography using 25 mM aqueous 15 NH4HCO3 solution and acetonitrile as eluents to obtain Example 261. HRMS calculated for C_5IH_4SC1FN₆O₉S: 974.2876, found: 975.2949 (M+H)

Example 262: l-|(ethoxycarbonyl)oxy|ethyl N-|(5S_a)-5-{3-chloro-2-metliyl-4-|2-(4methylpiperazin-Lyl)ethoxy|phenyl}-6-(4-fluorophenyl)thieno|2,3-7]pyrimidin-4-yl]2-{[2-(2-methoxyphenyI)pyrimidin-4-yl]methoxy}-D-phenylalaninate

Exaniple 263: l-[(dinicthylcarbamoyl)oxy|ethyl /V-[(5S_u)-5-{3-chloro-2-methyl-4-|2(4-methylpiperazin-l-yl)ethoxy]phenyl}-6-(4-fluorophenyl)thieno[2,3-7|pyriniidin-4ylJ-2-{|2-(2-methoxyphenyl)pyrimidin-4-yI|metlioxy}-D-phenylalaninatc

Example 264: 2V-[(55_fl)-5-{3-chloro-2-methyl-4-[2-(4-methylpipeiazin-l-yl)etlioxy| phenyl}-6-(4-fluorophenyl)thieno[2,3-i/]pyrimidin-4-yl]-2-({2-[3-(hydroxymethyl) phenyl]pyrimidin-4-yI}methoxy)-D-phenylalanine

Exaniple 265: A^r-|(5S_i/)-5-{3-chloro“2-methyl-4-[2-(4-methylpiperazin-l-yI)ethoxy]

ORIGINAL

- ISOphenyl}-6-(4-fluorophcnyl)thieno|2,3-rf|pyrimidin-4-yl]-2-({2-[2-(hydroxymethyl) pyridin-4-yl]pyrimidin-4-yl}methoxy)-D-phenylalanine

Example 266: 7V-|(55^, _u)-5-{3-chloro-2-methyl-4-[2-(4-methylpiperazin-l-yl)ethoxy] phenyl}-6-(4-fluorophenyl)thieno[2,3-rf|pyrimidin-4-yl]-2-({2-[6-(hydroxymethyl) pyridazin-4-yl|pyrimidin-4-yl}methoxy)-D-phenylalanine

Example 267: iV-|(55„)-5-{3-chIoro-2-methy l-4-[2-(4-methy lpiperazin-1 -y l)ethoxy ] phenyl}-6-(4-fluorophenyl)thieno[2,3-i/|pyrimidin-4-yl]-2-({2-|6-(hydroxymethyl) pyrazin-2-yl]pyrimidin-4-yl}methoxy)-D-phenylalanine

Example 268: Aⁱ-|(5A_ii)-5-{3-chloro-2-rnethyl-4-|2-(4-methylpiperazin-1-yl)cthoxyj phenyl}-6-(4-fluorophenyl)thieno[2,3-<f]pyrinudin-4-yl|-2-{[2'-(hydroxymethyl)-2,5'bipyrimidin-4-yI|methoxy}-D-phenylalanine

Example 269: 7V-[(55_a)-5-{3-chloro-2-methyl-4-[2-(4-niethylpiperazin-l-yl)ethoxy| phenyl}-6-(4-fluorophenyl)tliieno|2,3-rf] pyrimidin-4-yl|-2-({2-|4-(phosphonooxy) phenyl]pyrimidin-4-yl}methoxy)-D-plienylalanine

Example 270: 7V-[5-{3,5-dichloro-2,6-dimethyl-4-[2-(4-methylpiperazin-l-yl) ethoxy|phenyl}-6-(4-fluorophenyl)thieno[2,3-i/]pyrimidin-4-yl]-2-{|2-(2-methoxy phcnyl)pyrimidin-4-yl]methoxy}-D-phenylalanine

Step A: 4-brotno-2,6-dichloro-3,5-dimethyl-phenol

30.16 g 4-bromo-3,5-dimethyl-phenol (150 mmol) was dissolved in a mixture of 75 mL

1,2-dichloroethane and 75 mL acetonitrile, then 40.06 g NCS (300 mmol) was added portionwise and the mixture was stirred at r.t. until no further conversion was observed. Reaction mixture was concentrated under reduced pressure, the residue was dissolved in DCM, washed with water and brine. The organic layer was dried over NajSCL and concentrated under reduced pressure and used in the next step without further purification. ^fH NMR (400 MHz, DMSO-d₆): 10.10 (s, IH), 2.46 (s, 6H)

ORIGINAL

- I5l Step B: l-brotno-3.5-dichloro-4-tnethoxy-2,6-dimethyl~benzene

To a solution of 26.0 g 4-bromo-2,6-dichloro-3,5-dimethyl-phenol (96.3 mmol) and 26.6 g K₂CO₃ (192.6 mmol) in 300 mL MeCN 6.6 mL Mel (105.9 mmol) was added and the mixture was stirred at r.t. until no further conversion was observed. The solids were filtered off and the filtrate was concentrated under reduced pressure. The crude product was dissolved in DCM, washed with water and brine. The organic layer was dried over Na₂SO₄ and concentrated under reduced pressure and used in the next step wîthout further purification. 'H NMR (400 MHz, DMSO-d₆): 3.78 (s, 3H), 2.49 (s, 6H)

Step C: 2-(3,5-dichloro-4-methoxy-2,6-dimethyl-phenyl)-4,4.5,5-ietramethyl-I.3,210 dioxaborolane

10.0 g l-bromo-3,5-dichloro-4-methoxy-2,6-dimethyl-benzene (35.2 mmol) was dissolved in 360 mL dry THF under nitrogen and was cooled to -78 °C with dry ice-acetone.

23.2 mL nBuLi (L6 M in hexanes) (37.0 mmol) was added and the mixture was stirred for minutes, then 8.6 mL 2-isopropoxy-4,4,5,5-tetramethyl-l,3,2-dioxaborolane (42.24 mmol) was added and the mixture was allowed to warm up to r.t. It was quenched with brine, extracted with DCM, dried over Na2SO₄, filtered and concentrated under reduced pressure. The crude product was purified via flash chromatography using heptane and EtOAc as eluents to obtain 2-(3,5-dichloro-4-methoxy-2,6-dimethyl-phenyl)-4,4,5,5tetramethyl-l,3,2-dioxaborolane. ‘H NMR (400 MHz, DMSO-d₆): 3.81 (s, 3H), 2.33 (s,

6H). L34 (s, 12H)

Step D: ethyl 4~(3,5-dichloro-4-niethoxy-2,6-dimethyl~phenyl)thiophene-3-carboxylate

3.92 g ethyl 4-bromothiophene-3-carboxyiate (16.68 mmol) and 9.9 g 2-(3,5-dichloro-4methoxy-2,6-dimethyl-phenyI)-4,4,5,5-tetramethyl- 1,3,2-dioxaborolane (30.0 mmol) were dissolved in 140 mL dioxane, then 10,87 g CS2CO3 (33.36 mmol) dissolved in 40 mL water was added. Then 590 mg AtaPhos (0.83 mmol) was added, and the mixture was stirred under nitrogen at reflux température until no further conversion was observed. Then it was diluted with DCM and brine. After phase séparation the aqueous phase was extracted with DCM. The organic layers were combined and dried over Na₂SO₄, filtered and concentrated under reduced pressure. The crude product was purified via flash chromatography using

7^

ORIGINAL

- 152heptane and EtOAc as eluents to obtain ethyl 4-(3,5-dichloro-4-methoxy-2,6-dimethylphenyl)thiophene-3-carboxylate.

‘H NMR (400 MHz, DMSO-d₆): 8.53 (d, IH), 7.47 (d, IH), 4.02 (q, 2H), 3.83 (s, 3H), 1.95 (s, 6H), l .00 (t. 3H)

HRMS (Μ+ΝΕΕ)⁺ = 376.0538

Step E: ethyl 4-(3,5-dichloro-4-methoxy-2,6-dimethyl-phenyl)-2,5-diiodo-lhiophene-3carboxylate

65 g 4-(3,5-dichloro-4-methoxy-2,6-dimethyl-phenyl)thiophene-3-carboxylate (7.38 mmol) was dissolved in 75 mL acetonitrile, then 2.2 mL fluoroboric acid diethyl ether complex (16.23 mmol) and 3.65 g N-iodosuccinimide (l6.23 mmol) was added and the mixture was stirred at r.t. until no further conversion was observed. Reaction mixture was concentrated under reduced pressure, and the crude product was purified via flash chromatography using heptane and EtOAc as eluents to obtain ethyl 4-(3,5-dichloro-4methoxy-2,6-dimethyl-phenyl)-2,5-diiodo-thiophene-3-carboxylate. ’H NMR (400 MHz. DMSO-dû): 3.98 (q, 2H), 3.84 (s, 3H), 1.92 (s, 6H), 0.84 (t, 3H)

Step F: ethyl 4-(3,5-dichloro-4-methoxy-2,6-diniethyl-phenyl)-5-iodo-thiophene-3carhoxylate

5.29 g 4-(3,5-dichloro-4-methoxy-2,6-dimethy!-phenyl)-2,5-diiodo-thiophene-3carboxylate (8,66 mmol) was dissolved in 90 mL dry THF, then cooled to -78 °C under argon atmosphère. 6.7 mL isopropyl magnésium chloride, lithium chloride complex (1.3 M in THF) (8.66 mmol) was added and the mixture was stirred at -78 °C for 30 minutes. Then saturated aq. NH4CI was added and the mixture was extracted with ethyl acetate. The organic layer was dried over Na₂SÛ4 and concentrated under reduced pressure. The crude product was purified via flash chromatography using heptane and EtOAc as eluents to obtain ethyl 4-(3,5-dichloro-4-methoxy-2,6-diniethyl-phenyl)-5-iodo-thiophene-3carboxylate. ’H NMR (400 MHz, DMSO-d₆): 8.71 (s, IH), 4.01 (q, 2H), 3.86 (s, 3H), 1.89 (s, 6H), 0.99 (t, 3H)

Step G: ethyl 4-(3,5-dichloro-4-inethoxy-2.6-dimethyl-phenyl)-5-(4-fluorophenyl) thiophene-3-carboxylate

ORIGINAL

- 153 -

4.20 g ethyl 4-(3,5-dichloro-4-methoxy-2,6-dimethyl-phenyl)-5-iodo-thiophene-3carboxylate (8.66 mmol) and 1.82 g 4-fluorophenylboronic acid (13.0 mmol) were dissolved in 80 mL dioxane, then 5.64 g Cs?CO₃ (17.32 mmol) dissolved in 20 mL water was added. Then 500 mg Pd(PPh₃)4 (0.43 mmol) was added, and the mixture was stirred under nitrogen at 80 °C until no further conversion was observed. Then it was diiuted with DCM and brine. After phase séparation the aqueous phase was extracted with DCM. The organic layers were combined and dried over Na₂SO₄, filtered and concentrated under reduced pressure. The crude product was purified via flash chromatography using heptane and EtOAc as eluents to obtain ethyl 4-(3,5-dichloro-4-methoxy-2,6-dimethyl-phenyl)-5(4-fluorophenyl)thiophene-3-carboxylate.

'H NMR (400 MHz, DMSO-d₆): 8.58 (s, IH), 7.22-7.10 (m, 4H), 4.03 (q, 2H), 3.82 (s, 3H), 1.92 (s, 6H), l.OO (t, 3H)

HRMS (M+H)⁺ - 453.0498

Step H: ethyl 4-(3,5-dichloro-4-methoxy-2,6-dimethyl-phenyl)-5-(4-fluorophenyl)-2-nitrothiophene-3-carboxylate

1.97 g ethyl 4-(3,5-dichloro-4-methoxy-2,6-dimethyl-phenyl)-5-(4-fluorophenyl) thiophene-3-carboxylate (4.34 mmol) was dissolved in 40 mL dry acetonitrile, then 576 mg nitronium tetrafluoroborate (4.34 mmol) was added and the mixture was stirred at r.t. until no further conversion was observed. Then it was diiuted with DCM and brine. After phase séparation the aqueous phase was extracted with DCM. The organic layers were combined and dried over Na?SO4, filtered and concentrated under reduced pressure. The crude product was purified via flash chromatography using heptane and EtOAc as eluents to obtain ethyl 4-(3,5-dichloro-4-methoxy-2,6-dimethyl-phenyl)-5-(4-fluorophenyl)-2-nitrothiophene-3-carboxylate. ‘H NMR (400 MHz. DMSO-d₆): 7.37-7.33 (m, 2H), 7.32-7.26 (m, 2H), 4.14 (q, 2H), 3.82 (s, 3H), 2.06 (s, 6H), 0.88 (t, 3H)

Step I: ethyl 2-amino-4-(3,5-dichloro-4-niethoxy-2,6-dimethyl-phenyl)-5-(4-fluorophenyl) thiophene-3-carboxylate

1.85 g ethyl 4-(3,5-dichloro-4-methoxy-2,6-dimethyl-phenyl)-5-(4-fluorophenyl)-2-nitrothiophene-3-carboxylate (3.71 mmol) was dissolved in a mixture of 90 mL acetic acid and 18 mL water, then 2.43 g zinc dust (37.1 mmol) was added portionwise and the mixture

ORIGINAL

- I54was stirred at r.t. until no further conversion was observed. Réaction mixture was concentrated under reduced pressure, and the crude product was purified via flash chromatography using heptane and EtOAc as eluents to obtain ethyl 2-amino-4-(3,5dichloro-4-methoxy-2,6-dimethyl-phenyl)-5-(4-fluorophenyl)thiophene-3-carboxylate.

'H NMR (400 MHz, DMSO-d₆): 7.73 (s, 2H), 7.12-7.06 (m, 2H), 7.02-6.97 (m, 2H), 3.86-

3.80 (m, 2H), 3.80 (s, 3H), 2.01 (s, 6H), 0.72 (t, 3H)

HRMS (M+H)⁺= 456.0598

Step J: 5-(3,5-dichloro-4-methoxy’-2,6-dimethyl-phenyi)-6-(4-fliiorophenyl)-3W-thieno

[2,3-d]pyrimidin-4-one

l.l g ethyl 2-amino-4-(3,5-dichloro-4-rnethoxy-2,6-diniethyl-phenyl)-5-(4-fluoropheny!) thiophene-3-carboxylate (2.35 mmol) was dissolved in 20 mL formamide and it was stirred at 150 °C until no further conversion was observed. Then it was poured onto water and the precipitated product was collected by filtration to give 5-(3,5-dichloro-4-methoxy-2,6dimethyl-phenyl)-6-(4-fluorophenyl)-3H-thieno[2,3-d]pyrimidin-4-one.

‘H NMR (400 MHz, DMSO-d₆): 12.53 (br s, IH), 8.18 (s, IH), 7.23-7.16 (m, 4H), 3.84 (s, 3H), 1.96 (s, 6H)

HRMS (M+Hf = 449.0289

Step K: 4-chloro-5-(3.5-dichloro-4-methoxy-2,6-dimethyl-phenyl)-6-(4-fluorophenyl) thieno[2.3-d]pyrimidine

700 mg 5-(3,5-dichloro-4-rnethoxy-2,6-dimethyl-phenyl)-6-(4-fluorophenyl)-3//-thieno [2,3-i/]pyrimidin-4-one (1.56 mmol) was dissolved in 6 mL phosphorous oxychloride and it was stirred at 90 °C until no further conversion was observed. Reaction mixture was concentrated under reduced pressure, then to the crude product icy water was added and it was sonicated for 10 minutes. The precipitated product was collected by filtration to give 4-chloro-5-(3,5-dichloro-4-methoxy-2,6-diinethyl-phenyl)-6-(4-fluorophenyl)thieno[2,3-4/] pyrimidine.

'H NMR (400 MHz, DMSO-d₆): 9.02 (s, IH), 7.38-7.26 (m, 4H), 3.86 (s, 3H), 1.99 (s, 6H) HRMS (M+H/ = 466.9954

ORIGINAL

- 155 Step L: 2,6-dichloro-4-[4-chloro-6-(4-fluorophenyl)thieno[2,3-d]pyritnidin-â-ylJ-3,5dimethyl-phenol and 4-[4-bromo-6-(4-fliiorophenyl)thieno[2,3-à]pyrimidin-5-yl]-2,6dichloro-3,5-dimethyl-phenol

To a stirred solution of 700 mg 4-chloro-5-(3,5-dichloro-4-methoxy-2,6-dimethyl-phenyl)5 6-(4-fluorophenyl)thieno[2,3-i/]pyrimidine (1.50 mmol) in 15 mL DCM 3.0 mL boron tribromide (l M in DCM) (3.0 mmol) was added at 0 °C and the mixture was allowed to warm up to r.t. and it was stirred until no further conversion was observed. The mixture was quenched with saturated aq. NH4CI and extracted with DCM. The combined organic phases were dried over Na^SOg and concentrated under reduced pressure. The residue was I0 purified via flash chromatography using heptane and EtOAc as eluents to obtain 2,6dichloro-4-[4-chloro-6-(4-fluorophenyl)thieno[2,3-i/]pyrimidin-5-yl]-3,5-dnnethyl-phenol and 4-[4-bromo-6-(4-fluorophenyl)thieno[2,3-i/]pyrimidm-5-yl]-2,6-dichloro-3,5dimethyl-phenol as a 37:63 mixture of products.

‘H NMR (400 MHz, DMSO-d₆): 10.14 (br s, IH), 9.01 (s, IH), 7.40-7.23 (m, 4H), 1.95 (s, 15 6H) and 10.14 (br s, l H), 8.93 (s, l H), 7.40-7.23 (m, 4H), l .93 (s, 6H)

HRMS (M+H)⁺ = 452.9800 and 496.9287

Step M: 4-ch!oro-5-[3,5-dichloro-2.6-dimefhyl-4-[2-(4-melhylpiperazin-l-yl)ethoxy] phenyl]-6-(4-fluorophenyl)lhieno[2,3-d]pyrimidine and 4-bromo-5-[3.5-dichloro-2.6dimethyl-4-[2-(4-methylpiperazin-l-yl)ethoxy]phenyl]-6-(4-fluorophenyl)thieno[2,3-d] 20 pyrimidine

300 mg mixture of 2,6-dichloiO-4-[4-chloro-6-(4-fluorophenyl)thieno[2,3-c/]pyrimidin-5yl]-3,5-diinethyl-phenol and 4-[4-bromo-6-(4-fluorophenyl)thieno[2,3-i/]pyrimidin-5-yl]-

2.6- dichloro-3,5-dimethyl-phenol (0.62 mmol), 286 mg 2-(4-methylpiperazin-l-yl)ethanol (I.98 mmol) and 520 mg triphenyl phosphine (l.98 mmol) were dissolved in 10 mL dry toluene, then 460 mg di/erfbutyl azodicarboxylate (1.98 mmol) was added. The mixture was stirred at 50 °C under nitrogen until no further conversion was observed. The volatiles were evaporated under reduced pressure and the crude intermediate was purified via flash chromatography using EtOAc and methanol as eluents to obtain 4-chloro-5-[3,5-dichloro-

2.6- dimethyl-4-[2-(4-methylpiperazin-l-yl)ethoxy]phenyl]-6-(4-fluorophenyl)thieno[2,3-i/] 30 pyrimidine and 4-bromo-5-[3,5-dichloro-2,6-dimethyl-4-[2-(4-methylpiperazin-l-

ORIGINAL

- 156yl)ethoxy]phenyl]-6-(4-fluorophenyl)thieno[2,3-c/]pyrimidine as a 35:65 mixture of products.

^lH NMR (400 MHz, DMSO-d₆): 9.02 (S, IH), 7.40-7.22 (m, 4H), 4.H (t, 2H), 2.78 (t, 2H), 2.63-2.20 (m, 8H), 2.17 (br s, 3H), 1.98 (s, 6H) and 8.94 (S, IH), 7.40-7.22 (m, 4H), 4.11 (t, 2H), 2.78 (t, 2H), 2.63-2.20 (m, 8H), 2.15 (br s, 3H), 1.98 (s, 6H)

HRMS (M+H)⁺ = 579.0968 and 623.0455

Step N: Example 270

250 mg mixture of 4-chloro-5-[3,5-dichloro-2,6-dimethyl-4-[2-(4-methylpiperazin-lyl)ethoxy]phenyl]-6-(4-fluorophenyl)thieno[2,3-<7]pyrimidine and 4-bromo-5-[3,5dichloro-2,6-dimethyI-4-[2-(4-methylpiperazin-l-yl)ethoxy]phenyl]-6-(4-fiuorophenyl) thieno[2,3-i/]pyrimidine (0.41 mmol), 327 mg (2Æ)-2-amino-3-[2-[[2-(2-methoxy phenyl)pyrimidin-4-yl]methoxy]phenyl]propanoic acid (Préparation A4, 0.86 mmol) and 280 mg Cs₂CO₃ (0.86 mmol) was dissolved in 5 mL /er/-butanol and the mixture was stirred at 70°C until no further conversion was observed. The solids were filtered off and the filtrate was concentrated under reduced pressure. The crude product was purified via préparative reversed phase chromatography using 25 mM aqueous NH4HCO3 solution and MeCN, Example 270 was obtained. HRMS calculated for C48H46N7O5FSCI2: 921.2642, found: 461.6398 (M+2H)

Example 271: jV-[5-{2,6-dimethyl-4-[2-(4-methylpiperazin-l-yl)ethoxy]phenyI}-6-(4fluorophenyl)thieno|2,3-rf] pyriniidin-4-yl]-2-{[2-(2-methoxyphenyl)pyrimidin-4yl] methoxy }*D-phenylalanine

Example 272: jV-[(55„)-5-{3-chloro-2-methyl-4-[2-(4-methylpiperazin-l-yl)ethoxyl plienyl}-6-(4-iluorophenyl)thieno[2,3-i/lpyrimidin-4-yl]-P-(hydroxymethyl)-2-{[2-(2methoxyphcnyl)pyrimidin-4-yl]methoxy}phenylalanine

Example 273: /V-[(55_fl)-5-{3-chloro-2-methyl-4-|2-(4-methylpiperazin-l-yl)cthoxy] phenyl}-6-(4-fluoiOphenyl)thieno[2,3-r/| pyrimidin-4-yl|-p-hydroxy-2-{[2-(2-methoxy phenyl)pyrimidin-4-yl]methoxy}phenylalanine

ORIGINAL

- 157Example 274: jV-|(5S_a)-5-{3-chloro-2-methyI-4-[2-(4-methylpiperazin-l-yI)ethoxy] phenyl}-6-(4-fluorophenyl)thieno[2,3-rf]pyrimidin-4-yl]-p-(2-hydroxyethyl)-2-{[2-(2methoxyphenyI)pyrimidin-4-yl]methoxy}phenylalanine

ORIGINAL

- 158 PHARMACOLOGICAL STUDY

EXAMPLE A: Inhibition of Mcl-1 bv the fluorescence polarisation technique

The relative binding potency of each compound was determined via Fluorescence Polarisation (FP). The method utilised a Fluorescein labelled ligand (Fluorescein-pAlaAhx-A-REIGAQLRRMADDLNAQY-OH; mw 2,765) which binds to the Mcl-1 protein (such that Mcl-1 corresponds to the UniProtKB® primary' accession number: Q07820) leading to an increased anisotropy measured in milli-polarisation (mP) units using a reader. The addition of a compound which binds competitively to the same site as the ligand will resuit in a greater proportion of unbound ligand in the System indicated by a decrease in mP units.

Method l: An 11 point serial dilution of each compound was prepared in DMSO and 2 pl transferred into fiat bottomed. low binding, 384-well plate (final DMSO concentration 5%). 38μ1 of buffer (10 mM 4-(2-hydroxyethyl)-l-piperazineethanesulfonic acid [HEPES], 150 mM NaCl, 0.05 % Tween 20, pH 7.4), containing the Fluorescein labelled ligand (final concentration 1 nM) and Mcl-1 protein (final concentration 5 nM) was then added.

Assay plates were incubated ~2 hours al r.t. before FP was measured on a Biomek Synergy2 reader (Ex. 528 nm, Em. 640 nm, Cut off 510 nm) and mP units calculated. The binding of increasing doses of test compound was expressed as a percentage réduction in mP compared to a window eslablished between ‘5 % DMSO only’ and ‘100 % inhibition' Controls. 11-point dose response curves were plotted with XL-Fit software using a 4Parameter Logistic Model (Sigmoidal Dose-Response Model) and the inhibitory concentrations that gave a 50 % réduction in mP (IC50) were determined. Results obtained using Method 1 are presented in Table 1 below; IC50 of Mcl-1 inhibition obtained using Method 1 are not underlined.

ORIGINAL

- 159 Method 2t An 11 point serial dilution of each compound was prepared in DMSO and 2 μΐ transferred into fiat bottomed, low binding, 384-well plate (final DMSO concentration 5 %). 38 μΐ of buffer (20 mM Na₂FIPO4, ImM EDTA. 50 mM NaCl, pH 7.4), containing the Fluorescein labelled ligand (final concentration lOnM) and Mcl-l protein (final 5 concentration 10 nM) was then added.

Assay plates were incubated ~2 hours at r.t. before FP was measured on a Biomek Synergy2 reader (Ex. 528 nm, Em. 640 nm, Cut off 510 nm) and mP units calculated. The binding of increasing doses of test compound was expressed as a percentage réduction in mP compared to a window established between '5% DMSO only’ and ‘100% inhibition’ 10 Controls (50 μΜ unlabelled ligand). 11-point dose response curves were plotted with XLFit software using a 4-Parameter Logistic Model (Sigmoidal Dose-Response Model) and the inhibitory concentrations that gave a 50 % réduction in mP (ICjo) were determined. Results obtained using Method 2 are presented in Table 1 below; ICjoofMcl-i inhibition obtained using Method 2 are underlined.

The results show that the compounds of the invention inhibit interaction between the Mcl-l protein and the fluorescent peptide described hereinbefore.

EXAMPLE B: In vitro cvtotoxicitv

The cytotoxicity studies were carried out on the H929 multiple myeloma tumour line.

The cells are distributed onto microplates and exposed to the test compounds for 48 hours. The cell viability is then quantified by a colorimétrie assay, the Microculture Tétrazolium Assay (Cancer Res., 1987, 47. 939-942).

The results are expressed in IC50 (the concentration of compound that inhibits cell viability by 50 %) and are presented in Table 1 below.

The results show that the compounds of the invention are cytotoxic.

ORIGINAL

- 160 Table 1: ICso of Mcl-1 inhibition (fluorescence polarisation test) antl of cytotoxicity for H929 cells

Noie: IC50 ofMcl-1 inhibition obtained using Method 2 are underlined.

1--	lC»(gM)Mcl-l FF	IC»(M)MTT 11929		ICm (μΜ) Mcl-l FP	IC» (Μ) MTT H929
Example I	0.127	>6.0E-07	Example 29	0.063	2.52E-06
Example 2	56.9% @ 10 uM	>6.0E-07	Example 30	68.2% @ 10 uM	>3.75E-06
Example 3	0.1 14	3.18E-07	Example 31	16.7% @ 10 uM	>3.75E-06
Example 4	0.005	2.15E-07	Example 32	0.007	>3.75E-06
Example 5	ND	ND	Example 33	54.9% @ 10 μΜ	ND
Example 6	0.013	>6.0E-07	Example 34	0.726	4.27E-06
Example 7	0.008	I.38E-08	Example 35	24.78% @ 10 uM	>3.0E-05
Example 8	0.054	2.58E-07	Example 36	0.086	>7.50E-06
Example 9	2.697	>3.75E-06	Example 37	18.7% @ 10 11M	>3.75E-06
Example 10	72.75% @. 3.3 11M	>3.75E-06	Example 38	1.871	>3.75E-06
Example 11	35.55% @ 10 uM	>3.75E-06	Example 39	0.025	4.0IE-07
Example 12	0.014	6.47E-07	Example 40	0.006	3.28E-08
Example 13	58% @ 10 uM	>3.75E-06	Example 41	0.006	7.38E-09
Example 14	0.038	7.21E-07	Example 42	0.010	6.1E-08
Example 15	40.05% @ 10 uM	>3.75E-06	Example 43	0.006	1.25E-08
Example 16	0.006	9.93E-O8	Example 44	0.007	3.44E-09
Example 17	0.057	2.94E-07	Example 45	0.005	8.96E-10
Example 18	1.821	>1.14E-07	Example 46	0.024	1.66E-07
Example 19	0.020	I.75E-07	Example 47	0.008	8.39E-08
Example 20	0.026	7.86E-O8	Example 48	0.007	1.61 E-08
Example 21	0.006	3.97E-08	Example 49	0.003	4.I2E-09
Example 22	0.002	8.59E-09	Example 50	0.007	>l.50E-07
Example 23	41.27% @ 1 îiM	>6.0E-07	Example 51	0,005	2.33E-08
Example 24	0.008	4.73E-08	Example 52	0.301	ND
Example 25	55.5% @ 10 uM	>6.0E-07	Example 53	0.020	ND
Example 26	0.012	6.36E-09	Example 54	0.541	ND
Example 27	0.011	2.09E-09	Example 55	8.839	ND
Example 28	0.116	>1.88E-06	Example 56	0.019	ND

ORIGINAL

- I6l -

	IC_M (μΜ) Mcl-l FP	IC» (Μ) MÎT H929		IC_M (μΜ) Mcl-1 FP	IC_W (Μ) MTT H929
Example 57	0.106	ND	Example 88	1.225	ND
Example 58	52.8% @ 10 μΜ	ND	Example 89	72.13% @ 200 μΜ	ND
Example 59	0.127	ND	Example 90	0.023	ND
Example 60	0.092	ND	Example 91	6.372	ND
Example 61	0.036	ND	Example 92	12.887	ND
Example 62	0.060	ND	Example 93	0.080	ND
Example 63	12.18% @ 10 μΜ	ND	Example 94	6.901	ND
Example 64	0.025	ND	Example 95	0.077	ND
Example 65	0.345	>2.78E-06	Example 96	1.628	ND
Example 66	36.9% @ 10 μΜ	>3.75E-06	Example 97	0.073	ND
Example 67	2.079	ND	Example 98	2.873	ND
Examplc 68	0.142	ND	Example 99	28.3% @ 200 μΜ	ND
Example 69	0.109	ND	Example 100	4.747	ND
Example 70	0.974	ND	Example 101	0.462	ND
Examplc 71	39.015	ND	Example 102	0.073	ND
Example 72	12.007	ND	Example 103	0.715	ND
Examplc 73	0.276	ND	Example 104	0.046	ND
Example 74	0.621	ND	Example 105	0.704	ND
Example 75	0.280	ND	Example 106	0.065	ND
Example 76	3.177	ND	Example 107	0.544	ND
Example 77	75.45% @ 200 μΜ	ND	Example 108	0.079	ND
Example 78	0.135	ND	Example 109	1.858	ND
Example 79	0.429	ND	Example 110	0.839	ND
Example 80	0.487	ND	Example 111	4.452	ND
Example 81	20.241	ND	Example 112	0.104	ND
Example 82	1.109	ND	Example 113	0.045	ND
Example 83	0.258	ND	Examplc 114	0.981	ND
Example 84	4.022	ND	Example 115	1.753	ND
Example 85	0.228	ND	Examplc 116	1.059	ND
Example 86	9.976	ND	Example 117	2.603	ND
Example 87	62.6 % @ 7.4 μΜ	ND	Example 118	0.056	ND

ORIGINAL

- 162 -

	1C_M (μΜ) Mcl-i FP	ICjo (Μ) MÎT 11929		1C_W (μΜ) Mcl-l FP	IC_W(M)MTT H929
Example l ] 9	1.456	ND	Example 150	0.528	ND
Examplc 120	9.445	ND	Example 151	71.3% @66.7 μΜ	ND
Example I2l	0.167	ND	Example 152	0.047	ND
Example I22	10.215	ND	Example 153	75.7 % @ 50 μΜ	ND
Example 123	0.152	ND	Example 154	2.316	2.03 E-05
Example 124	75.7 % @ 50 μΜ	ND	Example 155	63.15 %@ 50 μΜ	ND
Example 125	13.710	ND	Example 156	0.309	ND
Example 126	41.2%@50μΜ	ND	Example 157	62.05 % @ 50 μΜ	ND
Example 127	0.913	ND	Example 158	0.104	ND
Example 128	10.722	ND	Example 159	1.770	ND
Example 129	0.053	ND	Example 160	11.725	ND
Example 130	67.4 % @ 50 μΜ	ND	Example 161	12.579	ND
Example 131	0.495	ND	Example 162	0.597	ND
Example 132	9.844	ND	Example 163	8.375	ND
Example 133	0.079	ND	Example 164	0.227	ND
Example 134	0.176	ND	Example 165	0.315	ND
Example 135	75.25% @ 50 μΜ	ND	Example 166	0.064	ND
Example 136	0.146	ND	Example 167	1.146	ND
Example 137	71.05% @ 50 μΜ	ND	Example 168	4.775	ND
Examplc 138	0.664	ND	Example 169	2.105	ND
Example 139	32.75% @ 10 μΜ	ND	Example 170	0.955	ND
Example 140	0.208	ND	Example 171	2.775	ND
Example 141	26.78% @ 10 uM	ND	Example 172	28.849	ND
Example 142	0.335	ND	Example 173	6.794	ND
Example 143	68.85% @ 50 μΜ	ND	Example 174	0.466	ND
Example 144	0.154	ND	Example 175	3.856	ND
Example 145	3.888	ND	Example 176	8.409	ND
Example 146	0.071	ND	Example 177	0.957	ND
Example 147	14.236	ND	Example 178	3.232	ND
Example 148	0.416	ND	Example 179	0.881	ND
Example 149	14.001	ND	Example 180	2.104	ND

ORIGINAL

-163 -

	IC» (μΜ) McJ-1 fp	IC» (Μ) MTT U929		IC_w(pM)Mcl-l fp	1(% (M) MTT H929
Example I8l	0.299	ND	Example 212	12.674	ND
Example 182	0.407	ND	Example 213	0.111	ND
Exampte 183	2.044	ND	Example 214	1.911	ND
Example 184	76.8% @ 200 μΜ	ND	Example 215	0.015	ND
Example I85	2.533	ND	Example 216	58.95% @ 10 μΜ	ND
Example 186	7.880	ND	Example 217	0.065	ND
Example 187	1.385	ND	Example 218	71.5% @50 μΜ	ND
Exampte 188	21.302	ND	Example 219	10.940	ND
Example I89	8.327	ND	Example 220	31.7% @ 10 μΜ	ND
Example I90	34.602	ND	Example 221	0.097	ND
Example i91	0.171	ND	Example 222	70% @ 50 μΜ	ND
Example 192	2.082	ND	Example 223	0.086	ND
Example 193	36.522	ND	Example 224	8.607	ND
Example 194	1.752	ND	Example 225	0.053	ND
Example 195	14.228	ND	Example 226	0.069	ND
Example 196	2.908	ND	Example 227	3.312	ND
Example 197	54.1%@200 μΜ	ND	Example 228	0.025	ND
Exampie 198	9.862	ND	Example 229	6.325	ND
Example 199	0.298	ND	Example 230	3.236	ND
Exampie 200	5.440	ND	Exampie 231	56.75% @ 50 μΜ	ND
Example 201	0.136	ND	Example 232	62.2% @ 50 μΜ	ND
Exampie 202	0.75 l	ND	Example 233	3.408	ND
Example 203	71.85% @ 100 μΜ	ND	Example 234	68.7% @ 50 μΜ	ND
Exampie 204	44.8% @ 100 μΜ	ND	Exampie 235	1.393	ND
Example 205	26.2% @ 100 μΜ	ND	Exampie 236	13.498	ND
Example 206	8.844	ND	Exampie 237	0.569	ND
Exampie 207	0.136	ND	Example 238	2.785	ND
Exampie 208	0.632	ND	Example 239	20.328	ND
Exampie 209	56.85% @ 200 μΜ	ND	Example 240	0.958	ND
Example 210	8.363	ND	Example 241	14.334	ND
Exampte 211	0.668	ND	Example 242	0.091	ND

ORIGINAL

- 164-

	IC_W (μΜ) Mcl-l FP	IC» (Μ) MTT 11929		ICwUiM) Mcl-l FP	IC_M (M) MTT H929
Example 243	0.073	ND	Example 259	1.347	>6.0E-07
Example 244	0.120	ND	Example 260	1.982	ND
Example 245	11.367	ND	Example 261	ND	ND
Exampie 246	0.293	> 1.50E-05	Example 262	ND	ND
Exampie 247	11.826	ND	Exampie 263	ND	ND
Exampie 248	2.176	ND	Exampie 264	ND	ND
Exampie 249	36.761	ND	Exampie 265	ND	ND
Exampie 250	0.617	ND	Exampie 266	ND	ND
Exampie 251	12.053	ND	Example 267	ND	ND
Exampie 252	0.223	ND	Example 268	ND	ND
Exampie 253	1.363	ND	Example 269	ND	ND
Exampie 254	22.08% @ 200 μΜ	ND	Exampie 270	0.055	5.22E-07
Example 255	41.08% @ 200 μΜ	ND	Exampie 271	ND	ND
Exampie 256	1.606	>6.0E-07	Example 272	ND	ND
Example 257	57.15% @ 10 uM	ND	Example 273	ND	ND
Example 258	1.070	4.28E-07	Example 274	ND	ND

ND: not determined

For partial inhibitors, the percentage fluorescence polarisation inhibition for a given concentration of the test compound is indicated. Accordingly, 45.1% @10 μΜ means that 45.1% fluorescence polarisation inhibition is observed for a concentration oftest compound equa! to 10 μΜ.

EXAMPLE C: Quantification of the cleaved form of PARP in vivo

The ability of the compounds ofthe invention to induce apoptosis, by measuring cleaved

PARP levels, is evaluated in a xenograft model of AMO-I multiple myeloma cells. y

1.10 AMO-l cells are grafted sub-cutaneously into immunosuppressed mice (SCID strain). 12 to 14 days after the graft, the animais are treated by intraveinous or oral routes with the various compounds. After treatment, the tumour masses are recovered and lysed, and the cleaved form of PARP is quantîfied in the tumour lysâtes.

The quantification is carried out using the Meso Scale Discovery (MSD) ELI SA platfomi test, which specifically assays the cleaved form of PARP. It is expressed in the

ORIGINAL

- 165 form of an activation factor corresponding to the ratio between the quantity of cleaved PARP in the treated mice divided by the quantity of cleaved PARP in the control mice.

The results (presented in Table 2 below) show that the compounds of the invention are capable of inducing apoptosis in AMO-l tumour cells in vivo.

Table 2: Quantification of the cleaved form of PARP in vivo

	PARP fokl		PARP fokl		PARP fold
Example 7	81.4	Example 43	II4.6	Example 51	52.2
Example 22	222.2	Example 44	S5.8	Example 256	99.5
Example 24	164.5	Example 45	I03.7	Example 258	132.3
Example 40	I I4.3	Example 49	138.8	Example 259	134.4

EXAMPLE D: Anti-tumour activity in vivo

The anti-tumour activity of the compounds of the invention is evaluated in a xenograft niodel of AMO-l multiple myeloma cells.

IxlO⁷ AMO-l cells are grafted sub-cutaneously into immunosuppressed mice (SCID strain).

to 8 days after the graft, when the tumour mass has reached about 150 mm³, the mice are treated with the varions compounds in a daily schedule (5-day treatment). The tumour mass is measured twice weekly from the start of treatment.

The compounds of the invention hâve anti-tumour activities (tumour régression) in the AMO-l multiple myeloma model with ΔΤ/C (qualification parameter of the activity of a product, which is measured by subtracting the médian tumor volume on the day of last treatment from the médian tumor volume on the day of first treatment / tumour volume of the untreated control group on the day of last treatment) ranging from -1.5 to -24.5 %. The results obtained show that the compounds of the invention induce significant tumour régression during the treatment period.

ORIGINAL

- 166EXAMPLE E: Pharmaceutical composition: Tablets

1000 tablets containing a dose of 5 mg of a compound selected from Examples l to 274..........5g

Wheat starch.....................................................................................................................20g

Maize starch............................................... 20g

Lactose............................. 30g

Magnésium stéarate............................................................................................................2g

Silica...................................................................................................................................Ig

Hydroxypropylcellulose................. 2g

ORIGINAL

- 167 -

Claims

1. Compounds of formula (I):

in which 1 is linked to the -NH- group and 2 is linked to the aromatic ring,

E represents a cycloalkyl group, a heterocycloaikyl group, an aryl group or a heteroaryl group,

X represents a nitrogen atom or a C-R4 group,

Y represents a nitrogen atom or a C-R3 group,

Ri represents a halogen atom, a linear or branched (C|-C_ft)alkyl group, a linear or branched (C₂-C6)alkenyl group, a linear or branched (Ci-Côjalkynyl group, a linear or branched (Cj-C^polyhaloalkyl group, a hydroxy group, a hydroxy(Ci-C6)alkyl group, a linear or branched (C[-C₆)alkoxy group, -S-(C[-C6)alkyl, a cyano group, a nitro group, -alkylfCo-Cej-NRçRçf, -O-alkyRCi-Cej-NRgRç’, -O-alkylfCi-Côj-Rio, -C(O)-OR₉, -O-C(O)-R₉, -C(O)-NR₉R₉’, -NR₉-C(O)-R₉’, -NR₉-C(O)-OR₉’, -alkyI(Ci-C₆)-NR₉-C(O)-R₉\ -SO₂-NR₉R₉’, -SO₃-alkyI(C₁-C₆),

R₂, R3, R4 and R5 independently of one another represent a hydrogen atom, a halogen atom, a linear or branched (C|-C&)alkyl group, a linear or branched

ORIGINAL

- 168 (C₂-Cè)alkenyl group, a linear or branched (C₂-C6)alkynyl group, a linear or branched (Ci-Côjpolyhaloalkyl group, a hydroxy group, a hydroxy(C|-C₆)alkyl group, a linear or branched (Ci-Cé)alkoxy group, -S-(C|-Cô)alkyl, a cyano group, a nitro group, -alkyl(Co-C6)-NR₉R₉’, -O-alkyl(C|-C6)-NR₉R₉’, -O-alkyl(C[-C₆)-Ri₀, 5 -C(O)-OR₉, -O-C(O)-R₉, -C(O)-NR₉R₉’, -NR₉-C(O)-R₉’, -NR₉-C(O)-OR₉’,

-alkyl(C[-C6)^_NR₉-C(O)-R₉', -SO₂-NR₉R9’, or -SO₂-alkyl(C]-C6), or the substituents of the pair (Ri, R₂) form together with the carbon atoms carrying them an aromatic or non-aromatic ring composed of from 5 to 7 ring members, which may contain from l to 3 heteroatoms selected from oxygen, sulphur and 10 nitrogen, it being understood that resulting ring may be substituted by from l to 2 groups selected from halogen, linear or branched (Ci-Cb)alkyl, -alkyl(Co-C6)-NR₉R₉\ -NRuRji’, -alkyl(Co-C6)-Cyi, or oxo,

R₆ represents a hydrogen atom, a halogen atom, a linear or branched (Cj-Côjalkyl group, a linear or branched (C₂-C6)alkenyl group, a linear or branched I5 (C₂-C6)alkynyl group, a linear or branched (Ci-Cô)polyhaloalkyl group, a hydroxy group, a linear or branched (C|-C₆)alkoxy group, -S-fCi-Côjalkyl, a cyano group, a nitro group, -alkyl(Co-C6)-NR₉R₉’, -O-Cyi, -alkyl(Co-C6)-Cyi, -alkenyl(C₂-C6)-Cyi, -alkynyl(C₂-C₆)-Cyi, -O-alkyl(C_rC₆)-R_l0, -C(O)-OR₉, -O-C(O)-R₉,

-C(O)-NR₉R₉’, -NR₉-C(O)-R₉’, -NR₉-C(O)-OR₉’, -alkyl(C_rC₆)-NR₉-C(O)-R₉’, 20 -SO₂-NR₉R₉’, or -SO₂-alkyl(C,-C₆),

R₇ represents a hydrogen atom, a linear or branched (Ci-Cs)alkyl group, a -CHR_aRb group, an aryl group, a heteroaryl group, an arylalkyl(C|-C₆) group, or a heteroarylalkyRCj-Cô) group,

Rx represents a linear or branched (Cj-Cf,)alkyl group, a linear or branched 25 (C₂-C₆)alkenyl group, a linear or branched (C₂-C6)alkynyl group, -Cy₂, a halogen atom, a cyano group, -C(O)-Rn, or -C(O)-NR||R]|’,

R₉ and R₉’ independently of one another represent a hydrogen atom, a linear or branched (Ci-C₆)alkyl group, or the substituents of the pair (R₉, R₉’) form together with the nitrogen atom 30 carrying them an aromatic or non-aromatic ring composed of from 5 to 7 ring members, which may contain in addition to the nitrogen atom from l to 3 heteroatoms selected from oxygen, sulphur and nitrogen, it being understood that

ORIGINAL

-169the nitrogen in question may be substituted by a group representing a hydrogen atom, or a linear or branched (Cj-Cbjalkyl group,

Rio represents -Cyj, -Cy3-alkyl(Co-C6)-Cy₄, -C(O)-NR₉R₉’, -NR9R9’, -OR₉, -NR₉-C(O)-R₉ ⁷, -O-alkyl(C_rC₆)-OR9, -SO2-R9, -C(O)-OR₉,or -NH-C(O)-NH-R₉, Ru and Ru’ independently of one another represent a hydrogen atom or an optionally substituted linear or branched (Ci-Côjalkyl group,

R12 represents a hydrogen atom, a hydroxy group, or a hydroxy(Cj-C6)aikyl group, R_a represents a hydrogen atom or a linear or branched (CpCojalkyl group,

Rb represents a -O-C(O)-O-R_c group, a -O-C(O)-NR_cR<;’ group, or a -O-P(O)(OR_C)2 group,

R_c and R_c’ independently of one another represent a hydrogen atom, a linear or branched (Ci-Cs)alkyl group, a cycloalkyl group, a (Ci-C₆)alkoxy(CrC₆)alkyl group, a (C|-C6)alkoxycarbonyl(C|-C₆)alkyl group, or the substituents of the pair (R_c, R<’) form together with the nitrogen atom carrying them a non-aromatic ring composed of from 5 to 7 ring members, which may contain in addition to the nitrogen atom from l to 3 heteroatoms selected from oxygen and nitrogen, it being understood that the nitrogen in question may be substituted by a group representing a linear or branched (Ci-Côjalkyl group,

Cyi, Cy₂, Cy3 and Cy₄ independently of one another, represent a cycloalkyl group, a heterocycloalkyl group, an aryl group or a heteroaryl group, n is an integer equal to 0, l or 2, it being understood that:

- “aryl” means a phenyl, naphthyl, biphenyl group, “heteroaryl” means any mono- or bi-cyclic group composed of from 5 to IO ring members, having at least one aromatic moiety and containing from l to 3 heteroatoms selected from oxygen, sulphur and nitrogen, “cycloalkyl” means any mono- or bi-cyclic non-aromatic carbocyclic group containing from 3 to IO ring members, “heterocycloalkyl” means any mono- or bi-cyclic non-aromatic carbocyclic group containing from 3 to IO ring members, and containing from l to 3 heteroatoms selected from oxygen, sulphur and nitrogen, which may include fused, bridged or

ORIGINAL

- I70spiro ring Systems, it being possible for the aryl, heteroaryl, cycloalkyl and heterocycloalkyl groups so defined and the alkyl, alkenyl, alkynyl, alkoxy groups, to be substituted by from l to 4 groups selected from optionally substituted linear or branched (C_rC₆)alkyl, optionally substituted linear or branched (C2-Cft)alkenyl, optionally substituted linear or branched (C2-C₆)alkynyl, optionally substituted linear or branched (C|-C₆)alkoxy, optionally substituted (Ci-CôJalkyî-S-, hydroxy, hydroxy(Cj-C₆)alkyI, oxo (or .V-oxide where appropriate), nitro, cyano, -C(O)-OR’, -O-C(O)-R’, -C(0)-NR’R”, -0-C(0)-NR’R”, -NR R , -(C=NR’)-OR”, -O-P(O)(OR’)2, -0-P(0)(0'M⁺)₂, linear or branched (Ci-Cù)polyhaloalkyl, trifluoromethoxy, halogen, or an aldohexose of formula:

in which each R’ is independent;

it being understood that R’ and R” independently of one another represent a hydrogen atom or an optionally substituted linear or branched (C_rC₆)alkyl group and M⁺represents a pharmaceutically acceptable monovalent cation.

their enantiomers, diastereoisomers and atropisomers, and addition salts thereof with a pharmaceutically acceptable acid or base.

2. Compound of formula (!) according to claim 1, wherein:

Ri and R₂ independently of one another represent a halogen atom, a linear or branched (C|-C₆)alkyl group, a hydroxy group, a hydroxy(Ci-C₆)alkyl group, a linear or branched (CpCJalkoxy group, or the substituents of the pair (Rj, R2) form together with the carbon atoms carrying them an aromatic ring composed of from 5 to 7 ring members, which may contain from 1 to 3 nitrogen atoms, it being understood that resulting ring may be

ORIGINAL

- I7l substituted by from l to 2 groups selected from halogen, linear or branched (C|-C₆)alkyl, or -alkyl(Co-C6)-NR₉R₉\

- R3 represents a hydrogen atom, a halogen atom, a linear or branched (C|-C₆)alkyi group, a hydroxy group, a linear or branched (C_rC₆)alkoxy group, or -O-alkyl(C|-C₆)-NR₉R₉’,

Ri and R5 independently of one another represent a hydrogen atom, a halogen atom, a linear or branched (Ci-C_b)alkyl group, a hydroxy group, a linear or branched (Ci-Cô)alkoxy group,

- R<, represents a hydrogen atom, a halogen atom, a linear or branched (C|-C₆)alkyl group, a linear or branched (Cj-Côjpolyhaloalkyl group, a hydroxy group, a linear or branched (Ci-C₆)alkoxy group, a cyano group, a nitro group. -alkyl(C₀-C₆)-NR₉R₉', -alkyl(C₀-C₆)-Cyi, -0-alkyl(C|-C₆)-Rio, or -C(O)-NR₉R₉’,

- R₇ represents a hydrogen atom, a linear or branched (Ci-C₈)alkyl group, a -CHR_aR_bgroup, or a heteroarylalkyl(C|-C₆) group,

- R₈ represents a linear or branched (C|-C₆)alkyl group, a linear or branched (C2-C₆)alkenyl group, a linear or branched (C₂-C₆)alkynyl group, -Cy₂, a halogen atom, or-C(O)-Rn,

R₉ and R₉’ independently of one another represent a hydrogen atom, or a linear or branched (C|-Cf,)alkyl group, or the substituents of the pair (R₉, R₉’) form together with the nitrogen atom carrying them a non-aromatic ring composed of from 5 to 7 ring members, which may contain in addition to the nitrogen atom from l to 3 heteroatoms selected from oxygen and nitrogen, it being understood that the nitrogen in question may be substituted by a group representing a linear or branched (C_rC₆)alkyl group,

- Rio represents -Cy₃ or -Cy₃-alkyl(Co-C6)-Cy₄,

R| i represents a linear or branched (Cj-C₆)alkyl group, it being possible for the aryl, heteroaryl, cycloalkyl and heterocycloalkyl groups so defined and the alky], alkenyl, alkynyl, alkoxy groups, to be substituted by from l to 4 groups selected from optionally substituted linear or branched (Ci-C₆)alkyl, optionally substituted linear or branched (C]-C₆)alkoxy, hydroxy, oxo (or A-oxide where appropriate), -C(O)-OR’, -C(O)-NR’R”, -O-C(O)-NR’R”, -NR'R”. -O-P(O)(OR’)₂,

ORIGINAL

- 172-O-P(O)(O’M )2, linear or branched (CrCôjpolyhaloalkyl, halogen. or an aldohexose of formula:

in which each R’ is independent; it being understood that R’ and R” independently of 5 one another represent a hydrogen atom or an optionally substituted linear or branched (C[-Cô)alkyl group and M⁺ represents a pharmaceutically acceptable monovalent cation.

3. Compounds according to claim l. wherein n is an integer equal to l.

4. Compounds according to claim l, wherein at least one the groups selected from R₂,

I0 R₃, R_| and R₅ does not represent a hydrogen atom.

5. Compounds according to claim l, wherein R.12 represents a hydrogen atom.

6. Compounds according to claim l. wherein R| represents a linear or branched (Cj-C₆)alkyl group or a halogen atom.

7. Compounds according to claim l, wherein R2 represents a linear or branched

I5 (C[-C₆)alkoxy group, a hydroxy group or a halogen atom.

8. Compounds according to claim l, wherein X represents a C-R4 group.

9. Compounds according to claim l, wherein Y represents a C-R3 group.

10. Compounds according to claim 1, wherein R₄ and R, represent a hydrogen atom.

11. Compound according to claim 1, wherein the substituents of the pair (R|, R₅) are

ORIGINAL

- 173identical and the substituents of the pair (R₂, R-t) are identical.

12. Compounds according to claim 1, wherein :

wherein R], R₂, R₉ and R₉’ are as defined in claim 1.

13. Compounds according to claim 1. wherein :

wherein R₉ and R₉’ are as defined in claim 1.

14. Compounds according to claim 1, wherein E represents a phenyi group, a pyridin-2-yl, a cyclohexyl group, a pyrazol-l-yl group, a cyclopentyl group, an indol-4-yl group, a

10 cyclopropyl group, a pyridin-3-yl group, an indoI-3-yl group, a naphth-l-yl group, an imidazol-4-yl group or a pyridin-4-yi group.

15. Compounds according to claim 1, which are compounds of formula (I-b):

ORIGINAL

- 174 -

wherein R_t, R₂, R_b, R_b, R₇, R₎₂, X, Y, A and n are as defined for formula (I).

16. Compounds according to claim l, wherein R_f, represents a hydrogen atom; a fluorine atom; a chlorine atom; a bromine atom; a methyl group; a trifluoromethyl group; a hydroxy group; a methoxy group; a linear (Ci-Cc)alkoxy group substituted by halogen atoms, a -C(O)-NR’R group or a -NR’R” group; a cyano; a nitro group; an aminomethyl group; a benzyl group; -0-alkyl(C|-C₆)-Rio; -C(O)-NR₉R₉\

17. Compounds according to claim 1, wherein R₇ represents a hydrogen atom, an optionally substituted linear or branched (Ci-Céjalkyl group, a -CHR^Rb group, or a heteroarylalkyl(Ci-Cé) group.

18. Compounds according to claim 1, wherein Rr represents a linear or branched (C₂-Cô)alkynyl group, an aryl group or a heteroaryl group.

19. Compounds according to claim 1, wherein R₉ and R₉’ independently of one another represent a linear or branched (Ci-C₆)alkyl group, or the substituents of the pair (R₉, R₉*) form together with the nitrogen atom carrying them a non-aromatic ring composed of from 5 to 7 ring members, which may contain in addition to the nitrogen atom from 1 to 3 heteroatoms selected from oxygen and nitrogen. it being understood that the nitrogen in question may be substituted by a linear or branched (C]-C₆)alkyl group.

20. Compounds according to claim 1, wherein R_t0 represents -Cy₃ or

ORIGINAL

-I75-Cy3-alkyl(Co-C₆)-Cy₄.

21. Compounds according to claim 20, wherein Cy₃ represents a cycloalkyl group, an aryl group or a heteroaryl group.

22. Compounds according to claim 20, wherein Cy₄ represents phenyl group or a 5 morpholinyl group.

23. Compounds according to claim 20, wherein

Rjo represents

in which p is an integer equal to 0 or 1 and R₁₅ represents a hydrogen atom, a hydroxy group, an optionally substituted linear or branched (C|-C&)alkyl group. a linear or branched (C,-C₆)alkoxy group, a -O-(CHR_I6-CHR|₇-O)q-R’ group, a -O-P(O)(OR’)₂ group, a -O-P(0)[O’M⁺)₂ group, a -O-C(O)-NR|₈R_]9 group, a di(Ci-C₆)aIkylamino(Cj-C₆)alkoxy group, a halogen atom, or an aldohexose of formula:

15 in which each R’ is independent;

it being understood that:

R’ represents a hydrogen atom or a linear or branched (C|-Cô)alkyl group,

- Ri6 represents a hydrogen atom or a (Cj-C₆)alkoxy(C|-C₆)alkyl group,

ORIGINAL

- 176 -

- R]7 represents a hydrogen atom or a hydroxy(C|-C₆)alkyl group,

- Ris represents a hydrogen atom or a (C|-C₆)alkoxy(C|-C₆)alkyl group,

- R_i9 represents a (C]-C6)alkoxy(Ci-C₆)alkyl group, a -(CH₂)_r-NR₉R₉’ group or a -(CH₂)_r-O-(CHR|₆-CHRi7-O)_q-R’ group,

5 - q is an integer equal to l, 2 or 3 and r is an integer equal to 0 or l,

- M⁺ represents a pharmaceutically acceptable monovalent cation.

24. Compounds according to claim 23, wherein the aldexose is D-mannose.

25. Compounds according to claim l, which are:

- À^f-[5-{3-chloro-2-methyl-4-[2-(4-methylpiperazin-l-yl)ethoxy]phenyl}-6-(4- ¹⁰ fluorophenyl)thieno[2,3-iZ]pyrimidin-4-yl]-2-[( 1 -methyl-1 Af-pyrazol-5-y 1) methoxy]-£)-phenylalanine,

- A^z-[5-{3-chloro-2-methyl-4-[2-(4-methylpiperazin-l-yl)ethoxy]phenyl}-6-(4fluorophenyl)thieno[2,3-i/]pyrimidin-4-yl]-2-[(2-ethoxypyrimidin-4-yl)methoxy]Ώ-phenyl alanine, ¹⁵ - AA[5-{3-chloro-2-methyl-4-[2-(4-methylpiperazin-1 -yl)ethoxy]phenyl }-6-(4fluorophenyl)thieno[2,3-i/]pyrimidin-4-yl]-2- {[2-(2-methoxyphenyl )pyrimidin-4yl]methoxy}-D-phenylalanine,

- ^[⁵-{3-chloro-2-methyl-4-[2-(4-methy]piperazin-l-yI)ethoxy]phenyl}-6-(iuran2-yl)thieno[2,3-c/]pyrimidin-4-y]]-2-methoxy-£)-phenylalanine, ²⁰ * ^J^-[5-{3-chloro-2-methyl-4-[2-(4-methylpiperazin-l-yl)ethoxy]phenyl}-6-(5nuorofuran-2-yl)thieno[2,3-i7]pyrimidin-4-yl]-2-methoxy-Z)-phenylalanine,

- /V-[5-{3-chloro-2-methyl-4-[2-(4-methylpiperazin-l -yl)ethoxy]phenyl}-6-(5fluorofuran-2-yl)lhieno[2,3-t7]pyrimidin-4-yl]-2-(2,2,2-trifluoroethoxy)-£>phenylalanine, ²⁵ - A^r-[5-{3-chIoro-2-methyl-4-[2-(4-methylpiperazin-l-yl)ethoxy]phenyl}-6-(5fluorofuran-2-yl)thieno[2,3-i7]pyrimidin-4-yl]-2-(pyridin-2-ylmethoxy)-£>phenylalanine,

- JV-[5- {3-chloro-2-methyl-4-[2-(4-methylpiperazin-1 -yi)ethoxy]phenyl} -6-(5 - fluorofuran-2-yl)thieno[2,3-i/]pyrimidin-4-yl]-2~[(l -methyl-17/-pyrazol-5-yl)

30 methoxy]-£)-phenylalanine,

ORIGINAL

- 177-

- jV-[5-{3-chloro-2-methyl-4-[2-(4-methylpiperazin-l-yl)ethoxy]phenyI}-6-(5fluorofuran-2-yl)thieno[2,3-i/]pyrimidin-4-yl]-2-[(l-ethyl-l//-pyrazol-5-yl) methoxy]-Z)-phenylalanine,

- A-[ 5- {3-chloro-2-methyl-4-[2-(4-methylpiperazin-1 -yl)ethoxy]pheny I} -6-(5fluorofuran-2-yl)thieno[2,3-i/]pyrimidÎn-4-yl]-2-[(2-ethoxypyrimidin-4-yl) methoxy]-D-phen ylalani ne,

- 2-[( l-butyl-l /7-pyrazoI-5-yl)methoxy]-7V-[5-{3-chloro-2-methyl-4-[2-(4-methyl piperazin-l-yl)ethoxy]phenyl}-6-(5-fIuorofuran-2-yl)thieno[2,3-i/]pyrimidin-4-yl]-

D-phenylalanine,

- /V-[5-{3-chloro-2-methyl-4-[2-(4-methylpiperazin-l-yl)ethoxy]phenyl}-6-(5fluorofuran-2-yl)thieno[2,3-i/]pyrimidin-4-yl]-2-{ [2-(2.2,2-trifluoroethoxy) pyrimidin-4-yl]methoxy}-£)-phenylalanine,

- A-[5-{3-chloro-2-methyl-4-[2-(4-methylpiperazin-l-yl)ethoxy]phenyl}-6-(5fluorofuran-2-yl)thieno[2,3-i/]pyrimidin-4-yl]-2-{[2-(2-methoxyphenyl)pyrimidin4-yI]methoxy} -D-phenylalanine,

- A^r-[5-{3-chloro-2-methyl-4-[2-(4-methylpiperazin-l-yl)ethoxy]phenyl}-6-(piOp-lyn-l-yl)thieno[2,3-i/]pyrimidin-4-yl]-2-methoxy-£)-phenylalanine,

- 2-[( l -tert-butyl-1 //-pyrazol-5-yl)mcthoxy 5- {3-chloro-2-methyl-4-[2-(4- niethylpipeiazin-l-yl)ethoxy]phenyl}-6-(prop-l-yn-l-yl)thieno[2,3-i/Jpyrimîdin-4yl]-£)-phenylalanine, A^f-[5-{3-chloro-2-methyl-4-[2-(4-methylpiperazin-l-yl)ethoxy]phenyl}-6-(prop-1yn-l-yi)thieno[2,3-<(|pyrimidin-4-yl]-2-{[2-(2-methoxyethyl)pyrimidin-4-yl] methoxy }-£>-phenylalanine,

- A'-] 5- {3-chloro-2-methyl-4-[2-(4-niethyipipcrazin-1 -yl)ethoxy]phenyI} -6-(prop-1 yn-l-yi)thieno[2,3-i/]pyrimidin-4-yl]-2-{[l-(2,2,2-trifluoroethyl)-l/7-pyrazol-5-yl] methoxy }-Z)-phenylalanine,

- N-[5-{3-chloro-2-methy 1 -4-[2-(4-methylpiperazin-1 -yl)ethoxy]phenyl} -6-(prop-1 yn-l-yI)thieno[2,3-i(|pyrimidin-4-yl]-2-{[2-(morpholin-4-yl)pyrjmidin-4-yl] methoxy }-Z)-phenylalanine,

- ^-[5- {3-chloro-2-methyl-4-[2-(4-methyipiperazin-1 -yl)ethoxy]phenyl} -6-(prop-1 yn-l-yl)thieno[2,3-c/]pyrimidin-4-yl]-2-{ [2-(2,2,2-trifluoroethoxy )pyrimidin-4-yl] methoxy}-Z)-pheny[aianine,

ORIGINAL

-I78-

- Λ-[5-{3-chloro-2-methyl-4-[2-(4-methylpiperazin-1 -yl)ethoxy]phenyl} -6-(prop-1 yn-l-yl)thieno[2,3-if]pyrimidin-4-yl]-2-{[2-(2-methoxyphenyl)pyrimidin-4-yl] methoxy }-Z)-phenylalanine,

- A-[5-{3-chloiO-4-[2-(dimcthylamino)cthoxy |-2-inethylphcnyl}-6-(prop-l-yn-l-yl) thieno[2,3-i7]pyrimidin-4-yl]-2-{[l-(2,2,2-trifluoroethyl)-I/7-pyrazol-5-yl] methoxy} -Ώ-phenylalanine,

- Y-[5-{3-chloro-4-[2-(dimethylamino)ethoxy]-2-methylphenyl)-6-(prop-l-yn-l-yl) thieno[2,3-i/]pyrimidin-4-yl]-2-{[2-(morpholin-4-yl)pyrimidin-4-yl]methoxy}-£)phenylalanine,

- ^-[5-{3-chloro-4-[2-(dimethylamino)ethoxy]-2-methylphenyl}-6-(prop-1 -yn-1 -yl) thieno[2,3-rf]pyrimidin-4-yl]-2-{[2-(2.2,2-trifluoroethoxy)pyrimidin-4-yl] methoxy} -D-phenylalanine,

- jV-[5-{3-chloro-4-[2-(dimethylamino)ethoxy]-2-melhylphenyl}-6-(prop-l-yn-l-yl) thieno[2,3-t/]pyrimidin-4-yl]-2-{[2-(2-methoxyphenyI)pyrimidin-4-yl] methoxy }-Dphenylalanine,

- 7V-[5-{3-chloro-4~[2-(dimethylamino)ethoxy]-2-methylphenyl}-6-(4-fluoro phenyl)thieno[2,3-i/]pyrimidin-4-yl]-2-({2-[2-(2-methoxyethoxy)phenyl]pyrimidin4-yl}methoxy)-£)-phenylalanine;

- ethyl N-[(5S_a)-5-{3-chIoro-2-methyl-4-[2-(4-methylpiperazin-l-yl)ethoxy] phenyl }-6-(4-fluorophenyl)thieno[2,3-iZ]pyrimidîn-4-yl]-2-{[2-(2-methoxyphenyl) pyrimîdin-4-yl]methoxy}-D-phenylaIaninate;

- ethyl jV-[(55_a)-5-{3-chloro-2-methyl-4-[2-(4-methylpiperazin-l-yl)ethoxy] phenyl }-6-(prop-l-yn-l-yl)thieno[2,3-</]pyrirnidm-4-yl]-2-{[2-(2-methoxyphenyl) pyrimidin-4-yl]methoxy}-Z)-phenylalaninate;

- ethyl Y-[(55_a)-5-{3-chloro-4-[2-(dimethylamino)ethoxy]-2-methylphenyl}-6(prop-l-yn-l-yl)thieno[2,3-d]pyrimidin-4-yl]-2-{[2-(2-methoxyphenyl)pyrimidin4-yl]methoxy)-D-phenylalaninate;

- jV-[5-{3,5-dichloro-2,6-dimethyl-4-[2-(4-methylpiperazin-l-yl) ethoxy]phenyl)-6-(4-fluorophenyl)thieno[2,3-i/]pyrimidin-4-yl]-2-{[2-(2methoxyphenyl)pyrimidin-4-yl]methoxy}-D-pheny[alanine.

26. Process for the préparation of a compound of formula (I) according to claim 1,

ORIGINAL

- I79characterised in that there is used as sterling materiai the compound of formula (Il-a):

Cl^/A\ (H-a) wherein Z represents bromine or iodine and A is as defined for formula (I) in which 1 is linked to the chlorine atom and 2 is linked to the Z group, which compound of formula (ΙΙ-a) is subjected to coupling with a compound of formula (III):

R_i2 (III) wherein R₆. R|₂, E and n are as defined for formula (1), and Alk represents a linear or branched (C]-C₆)alkyl group,

10 to yield the compound of formula (IV):

Alk

^R12 wherein R₆, R₁₂, A, E and n are as defined for formula (I) and, Z and Alk is as defined before.

compound of formula (IV) which îs further subjected to coupling with compound of

15 formula (V):

ORIGINAL

- 180-

B

Κβ2θ ORjjj (V) wherein R|, R₂, R₅, X and Y are as defined for formula (I), and R_Bl and R_B2 represent a hydrogen atom, a linear or branched (C]-C₆) alkyl group, or R_B| and R_B2 form with the oxygen carrying them an optionally methylated ring, to yîeld the compound of formula (VI):

(VI) wherein R|, R₂, R₅, R₆, R₍₂, X, Y, A. E and n are as defined for formula (!) and Alk is as defined before, the Alk-O-C(O)- ester fonction of which compound of formula (VI) is hydrolysed to I0 yield the carboxylic acid, which may optionally be reacted with an alcohol of formula

R₇’-OH or a chlorinated compound of formula R/-C1 wherein R₇' represents a linear or branched (Ci-Cg)alkyl group, a -CHR_aRb group, an aryl group, a heteroaryl group, an arylalkyl(C|-C₆) group, or a heteroarylalkyl(Ci-C₆) group, R_a and R_b are as defined for formula (l),

15 to yield the compound of formula (I), which may be purified according to a conventional séparation technique, which is converted, if desired, into its addition salts with a pharmaceutically acceptable acid or base and which is optionally separated into

ORIGINAL

- I8l îts isomers according to a conventional séparation technique, it being understood that at any moment considered appropriate during the course of the process described above, some groups (hydroxy, amino...) of the starting reagents or of the synthesis intermediates can be protected, subsequently deprotected and 5 functionalized, as required by the synthesis.

27. Process for the préparation of a compound of formula (I) according to claim l. characterised in that there is used as starting material the compound of formula (ll-b):

c+'S fli-b) wherein A is as defined in formula (I) in which 1 is linked to the chlorine atom and 2 10 is linked to the iodine atom.

which compound of formula (ΙΙ-b) is subjected to coupling with a compound of formula (V):

(V) wherein Rj, R₂, R₅, X and Y are as defined for formula (1), and R_B| and R_B2 represent

5 a hydrogen atom, a linear or branched (C_rC₆) alkyl group, or R_B, and R_B2 form with the oxygen carrying them an optionally methylated ring, to yield the compound of formula (VII):

(VII)

ORIGINAL

- 182wherein R|, R₂, R5, A, X and Y are as defined in formula (I), which compound of formula (VII) is further subjected to coupling with a compound of formula (III):

Alk

R.2 (III) wherein R₆, R|₂, E and n are as defined for formula (I), and Alk represents a linear or branched (C|-C₆)alkyl group, to yield the compound of formula (VI):

(VI) wherein R|, R₂, R>, R,₂, X, Y, A, E and n are as defined for formula (1) and Alk is

10 as defined before, the Alk-O-C(O)- ester function of which compound of formula (VI) is hydrolysed to yield the carboxylic acid, which may optionally be reacted with an alcohol of formula R₇’-OH or a chlorinated compound of formula R₇’-C1 wherein R₇’ represents a linear or branched (Ci-Cgjalkyl group, a -CHR_aRh group, an aryl group, a heteroaryl group, an arylalkyl(Ci-C₆) group, or a heteroarylalkyl(Ci-C₆) group, R_a and R_h are as defined for formula (I),

ORIGINAL

- 183to yield the compound of formula (I), which may be purified according to a conventional séparation technique, which is converted. if desired, into its addition salts with a pharmaceutically acceptable acid or base and which is optionally separated into its isomers according to a conventional séparation technique, it being understood that at any moment considered appropriate during the course of the process described above, some groups (hydroxy, amino...) of the starting reagents or of the synthesis intermediates can be protected, subsequently deprotected and functionalized, as required by the synthesis.

28. Pharmaceutical composition comprising a compound of formula (I) according to any one of daims l to 25 or an addition sait thereof with a pharmaceutically acceptable acid or base in combination with one or more pharmaceutically acceptable excipients.

29. Pharmaceutical composition according to claim 28 for use as pro-apoptotic agents.

30. Pharmaceutical composition according to claim 29 fbr use in the treatment of cancers and of auto-immune and immune System diseases.

31. Pharmaceutical composition according to claim 30 for use in the treatment of cancers of the bladder, brain, breast and utérus, chronic lymphoid leukaemias, cancer of the colon, œsophagus and liver, lymphoblastic leukaemias, acute myeloid leukaemias, lymphomas, melanomas, malignant haemopathies, myelomas, ovarian cancer, nonsmall-cell lung cancer, prostate cancer, pancreatic cancer and small-cell lung cancer.

32. Use of a pharmaceutical composition according to claim 28 in the manufacture of médicaments for use as pro-apoptotic agents.

33. Use of a pharmaceutical composition according to claim 28 in the manufacture of médicaments fbr use in the treatment of cancers and of auto-immune and immune System diseases.

34. Use of a pharmaceutical composition according to claim 28 in the manufacture of

ORIGINAL

- I84 médicaments for use in the treatment of cancers of the bladder, brain. breast and utérus, chronic lymphoid leukaemias, cancer of the colon, œsophagus and liver, lymphoblastic leukaemias, acute myeloid leukaemias, lymphomas, melanomas, malignant haemopathies, myelomas, ovarian cancer, non-small-cell lung cancer, prostate cancer, pancreatic cancer and small-cell lung cancer.

35. Compound of formula (I) according to any one of daims l to 25, or an addition sait thereof with a pharmaceutically acceptable acid or base, for use in the treatment of cancers of the bladder, brain, breast and utérus, chronic lymphoid leukaemias, cancer of the colon, œsophagus and liver, lymphoblastic leukaemias, acute myeloid leukaemias, lymphomas, melanomas, malignant haemopathies, myelomas, ovarian cancer, non-small-cell lung cancer, prostate cancer, pancreatic cancer and small-cell lung cancer.

36. Use of a compound of formula (I) according to any one of daims l to 25, or an addition sait thereof with a pharmaceutically acceptable acid or base, in the manufacture of médicaments for use in the treatment of cancers of the bladder, brain, breast and utérus, chronic lymphoid leukaemias, cancer of the colon, œsophagus and liver, lymphoblastic leukaemias, acute myeloid leukaemias, lymphomas, melanomas, malignant haemopathies, myelomas, ovarian cancer, non-small-cell lung cancer, prostate cancer, pancreatic cancer and small-cell lung cancer.

37. Combination of a compound of formula (l) according to any one of daims l to 25 with an anti-cancer agent selected from genotoxic agents, mitotic poisons, anti-metabolites, protéasome inhibitors, kinase inhibitors and antibodies.

38. Pharmaceutical composition comprising a combination according to daim 37 in combination with one or more pharmaceutically acceptable excipients.

39. Combination according to claim 37 for use in the treatment of cancers.

40. Use of a combination according to claim 37 in the manufacture of médicaments for

ORIGINAL

- I85use in the treatment of cancers.

41. Compound of formula (I) according to any one of claims 1 to 25 for use in the treatment of cancers requiring radiotherapy.