EA044387B1 - G12C KRAS INHIBITORS AND METHODS OF THEIR APPLICATION - Google Patents

Description

Field of technology

Provided herein are G12C KRAS inhibitors, their pharmaceutically acceptable salts, atropisomers, a pharmaceutical composition based thereon, and methods of using them. Such inhibitors are useful for the treatment of a number of disorders, including pancreatic cancer, colorectal cancer and lung cancer.

State of the art

KRAS gene mutations are common in pancreatic cancer, lung adenocarcinoma, colorectal cancer, gallbladder cancer, thyroid cancer, and bile duct cancer. KRAS mutations are also observed in approximately 25% of patients with NSCLC, and some studies indicate that KRAS mutations are a negative prognostic factor in patients with NSCLC. Recently, mutations in the Kirsten rat sarcoma viral oncogene V-Ki-ras2 homolog (KRAS) homolog were found to confer resistance to epidermal growth factor receptor (EGFR)-targeting therapies in colorectal cancer; Accordingly, KRAS mutational status may provide important information before initiating TKL therapy. Taken together, there is a medical need for new ways to treat patients with pancreatic cancer, lung adenocarcinoma, or colorectal cancer, especially for those who have been diagnosed with such cancers. characterized by a KRAS mutation, including those who progressed after chemotherapy.

Short description

This document provides a connection selected from:

or a pharmaceutically acceptable salt thereof.

In another embodiment, provided herein is a pharmaceutically acceptable salt of a compound, wherein said compound is

In another embodiment, a connection is provided herein, wherein said connection is

In another embodiment, provided herein is a pharmaceutically acceptable salt of atropoisomer 1 of compound. J

N - _s ' FF n An A An' / he ή ) \ I A'' corresponding to the 1st elution isomer, which can be obtained by separating atropisomers using supercritical liquid chromatography using Chiralpak IC, 30x250 mm, 5 µm, 55% MeOH/CO2, 120 g/min, 10.3 MPa (103 bar).

In another embodiment, provided herein is a pharmaceutically acceptable salt of atropoisomer 2 of compound

corresponding to the 2nd elution isomer, which can be obtained by separating atropisomers by supercritical liquid chromatography using Chiralpak IC, 30x250 mm, 5 µm, 55% MeOH/^, 120 g/min, 10.3 MPa (103 bar).

In another embodiment, atropisomer 1 of compound A ⁰ is provided herein

corresponding to the 1st elution isomer, which can be obtained by separating atropisomers by supercritical liquid chromatography using Chiralpak IC, 30x250 mm, 5 µm, 55% MeOH/^, 120 g/min, 10.3 MPa (103 bar).

In another embodiment, atropisomer 2 of the compound is provided herein

corresponding to the 2nd elution isomer, which can be obtained by separating atropisomers by supercritical liquid chromatography using Chiralpak IC, 30x250 mm, 5 µm, 55% MeOH/CO2, 120 g/min, 10.3 MPa (103 bar).

In another embodiment, provided herein is a pharmaceutically acceptable salt of a compound, wherein the compound is

In another embodiment, provided herein is a pharmaceutically acceptable salt of a compound, wherein the compound is

A

- 2 044387

In another embodiment, a connection is provided herein, where the connection is

In another embodiment, a connection is provided herein, where the connection is a

In another embodiment, provided herein is a pharmaceutical composition for the treatment of cancer comprising a compound of the invention and a pharmaceutically acceptable excipient.

In another embodiment, provided herein is an in vitro method of inhibiting KRAS G12C in a cell, comprising contacting the cell with a compound of the invention or a composition of the invention.

In another embodiment, provided herein is a method of treating cancer in a subject in need of the treatment, comprising administering to the subject a therapeutically effective amount of a compound of the invention or a composition of the invention.

In another embodiment of a method of treating cancer in a subject, the cancer is non-small cell lung cancer, colorectal cancer, pancreatic cancer, appendix cancer, endometrial cancer, stomach cancer, small bowel cancer, nasal cancer, bile duct cancer, or skin cancer.

In another embodiment of a method of treating cancer in a subject, the cancer is non-small cell lung cancer.

In another embodiment of a method of treating cancer in a subject, the cancer is pancreatic cancer.

In another embodiment of a method of treating cancer in a subject, the cancer is colorectal cancer.

In another embodiment of a method of treating cancer in a subject, the cancer is mediated by an O12C mutation of KRAS.

In another embodiment, the connection is In another embodiment, the connection is In another embodiment, the connection is This document provides a connection where the specified hu This document provides a connection where the specified Cl x5cc ^F o-^n-^n^ci Hu This document provides a connection where the specified A ΧλΧ' Au

- 3 044387

In another embodiment, a connection is provided herein, wherein said connection is

In another embodiment, a connection is provided herein, wherein said connection is

Detailed Description Definitions

The following abbreviations and abbreviations are used in this document.

AcOH acetic acid drive or drive. water VOS or Vos tert-butyloxycarbonyl srte cyclopentyl methyl ether DCE 1,2-dichloroethane DABCO 1,4-diazabicyclo[2.2.2]octane DCM dichloromethane DMA N,N-dimethylacetamide DMAP 4-dimethylaminopyridine DME 1,2-dimethoxyethane DMF N,N-dimethylformamide DMSO dimethyl sulphate oxide Dppf, DPPF or dppf 1,G-bis(diphenylphosphino)ferrocene eq or eq. or equivalent equivalent ESI or ES electrospray ionization Et ethyl

- 4 044387

Et ₂ O diethyl ether EtOAc ethyl acetate G gram h. hour HPLC high pressure liquid chromatography iPr isopropyl iPr ₂ NEt or DIPEA N-ethyldiisopropylamine (Hunig's base) KHMDS potassium hexamethyldisilazide KOAc potassium acetate Lawsson's reagent 2,4-bis(4-methoxyphenyl)-2,4-dithioxo-1,3,2,4dithiadiphosphetane, 2,4-bis-(4-methoxyphenyl)-1,3dithia-2,4-diphosphetane-2,4 -disulfide LC MS, LCMS, LC-MS or LC/MS mass spectrometry with liquid chromatography LG leaving group (e.g. halogen, mesylate, triflate) LHMDS or LiHMDS lithium hexamethyldisilazide mass/charge mass divided by charge Me methyl MeCN acetonitrile MeOH methanol Met metal particles intended for cross-coupling reactions (for example, MgX, ZnX, SnR ₃ , SiR ₃ , B(OR) ₂ ) MG milligram min. minutes ml milliliters MS mass spectra NaHMDS sodium hexamethyldisilazide N.B.S. N-bromosuccinimide n-BuLi n-butyllithium NCS N-chlorosuccinimide NMR nuclear magnetic resonance Pd ₂ (dba) ₃ tris(dibenzyl indenacetone)dipalladium(0)

-5044387

[1,1- Pd(dppf)Cl ₂ DCM bis(diphenylphosphino)ferrocene]dichloropalladium(P), complex with dichloromethane Pd(PPh ₃ ) ₄ tetrakis(triphenylphosphine)palladium(0) Ph phenyl PR or PG or def. group protective group rbf round bottom flask RP-HPLC reverse phase high pressure liquid chromatography RT or k.t. room temperature us. or sat. saturated SFC supercritical fluid chromatography (2-dicyclohexylphosphino-2',6'-dimethoxybiphenyl) SPhos Pd G3 or SPhos G3 [2-(2-amino-1,1-biphenyl)] palladium(P) methanesulfonate TBAF tetra-N-butylammonium fluoride TBTU Ν, Ν, Ν',U-tetramethyl-O-(benzotriazole-1- yl)uronium tetrafluoroborate /BuOH /u/?e/i-butanol TEA or Et ₃ N trimethylamine T.F.A. trifluoroacetic acid THF tetrahydrofuran UV ultraviolet

The use of the singular, plural, and similar references in the context of teaching the present invention (especially in the context of the claims) should be construed to include both singular and plural forms unless otherwise indicated. The listing of ranges of values in this document is intended only to serve as a shorthand way of referring individually to each individual value falling within a given range unless otherwise noted herein, and each individual value is included in the description as if it were separate. specified in this document. The use of any and all examples or illustrative language (eg, such as) presented herein is intended to better illustrate the present invention and is not intended to limit the scope of the present invention unless otherwise stated. Nothing in the specification should be construed as indicating any unstated element as essential to the practice of the present invention.

This document provides a connection selected from:

or a pharmaceutically acceptable salt thereof.

- 6 044387

In another embodiment, provided herein is a pharmaceutically acceptable salt of a compound, wherein said compound is

In another embodiment, a connection is provided herein, wherein said connection is

In another embodiment, provided herein is a pharmaceutically acceptable salt of atropoisomer 1 of compound

corresponding to the 1st elution isomer, which can be obtained by separating atropisomers using supercritical liquid chromatography using Chiralpak IC, 30x250 mm, 5 µm, 55% MeOH/CO ₂ , 120 g/min, 10.3 MPa (103 bar).

In another embodiment, provided herein is a pharmaceutically acceptable salt of atropoisomer 2 of compound

corresponding to the 2nd eluting isomer, which can be obtained by separating atropisomers using supercritical liquid chromatography using Chiralpak IC, 30x250 mm, 5 µm, 55% MeOH/CO ₂ , 120 g/min, 10.3 MPa (103 bar).

In another embodiment, atropisomer 1 of the compound is provided herein

corresponding to the 1st elution isomer, which can be obtained by separating atropisomers using supercritical liquid chromatography using Chiralpak IC, 30x250 mm, 5 µm, 55% MeOH/CO ₂ , 120 g/min, 10.3 MPa (103 bar).

In another embodiment, atropisomer 2 of the compound is provided herein

- 7 044387 corresponding to the 2nd elution isomer, which can be obtained by separating atropisomers using supercritical liquid chromatography using Chiralpak IC,

30x250 mm, 5 µm, 55% MeOH/CO ₂ , 120 g/min, 10.3 MPa (103 bar).

In another embodiment, provided herein is a pharmaceutically acceptable salt of a compound, wherein the compound is

In another embodiment, provided herein is a pharmaceutically acceptable salt of a compound, wherein the compound is

IN ^=/

In another embodiment, a connection is provided herein, where the connection is

In another embodiment, a connection is provided herein, where the connection is

In another embodiment, provided herein is a pharmaceutical composition for the treatment of cancer comprising a compound of the invention and a pharmaceutically acceptable excipient.

In another embodiment, provided herein is an in vitro method of inhibiting KRAS G12C in a cell, comprising contacting the cell with a compound of the invention or a composition of the invention.

In another embodiment, provided herein is a method of treating cancer in a subject in need of the treatment, comprising administering to the subject a therapeutically effective amount of a compound of the invention or a composition of the invention.

In another embodiment of a method of treating cancer in a subject, the cancer is non-small cell lung cancer, colorectal cancer, pancreatic cancer, appendix cancer, endometrial cancer, stomach cancer, small bowel cancer, nasal cancer, bile duct cancer, or skin cancer.

In another embodiment of a method of treating cancer in a subject, the cancer is non-small cell lung cancer.

In another embodiment of a method of treating cancer in a subject, the cancer is pancreatic cancer.

In another embodiment of a method of treating cancer in a subject, the cancer is colorectal cancer.

In another embodiment of a method of treating cancer in a subject, the cancer is mediated by a G12C mutation of KRAS.

- 8 044387

In another embodiment, the connection is a connection provided herein, wherein the specified

In another embodiment, a connection is provided herein, wherein said connection is

C.I.

In another embodiment, a connection is provided herein, wherein said connection is

In another embodiment, a connection is provided herein, wherein said connection is

In another embodiment, a connection is provided herein, wherein said connection is

The following Examples 1-11 are labeled using a classification system in which the first number refers to the method used to synthesize the compound, the second number is an identification number, and the third number, if present, refers to the order of elution of the compound in the chromatographic separation method. If the third number is missing, the compound is a single compound or a mixture of isomers. Examples 12-53 use a classification system in which the first number is an identification number and the second number, if present, refers to the order of elution of the compound in the chromatographic separation method. If the second number is missing, the compound is a single compound or a mixture of isomers. The sequential numbering of examples is interrupted and certain example numbers are deliberately omitted for formatting reasons. - means no changes have been made or there are no entries in the corresponding field. Particularly provided connections include those listed in table. 1 and 1(a).

- 9 044387

Table 1

No. approx. Chemical structure No. approx. Chemical structure No. note Chemical structure 1-1 \\ about N - , --N ь ^; t ': ^{Ivg N} P\ 1 CH 1-6 M' ⁰ N-^ N tin 1-11 . o κ,Ζ / And..... “O x=A^ 1-2 1-3 V A C - CI V' I ¹ ' ¹ -Nil N-. A 1 C S - V ί 1 f ¹¹ . ¹ yen 1-7 1-8 °·? And A _α is it 0? ' γ*\ ' - Ν A JI cl N G G1 - ''ON □ x a m - / / _-Ν _ Α, Ν ci G 1 ι: a-· g _he 1-12 1-13 Well, /'-N GI A ?' N J ί Cl'| F Γι l - - an O n.A~, .g .J ι) ,- S eleven 1! 1 LI F Γί -' - ' He o i N, 1 > -N . Cl A A/ 0 1

- 10 044387

- 11 044387

1-16 about ns G . '—N -S //N.J. II 1 L - F । I ΐ 'ΌΗ 9 hY” .. -N ΐ· Ν 11 1 LI F ή He 1- 19-2 1 0=/ GN 2-7 O X r* .- N Λ f Ί oY '/ 'f LJ ' IL - OH o .V \ - --N f [Г* i ¹ N [ 1 U ' TF j L 1 II '''... ..OH o ” to -- , / / ' ,- N / V ^s .1( N or y“ T A ό ^: - -· OH 1-17 4V A n n-Y ⁴ । '—Ν' AX ₍ i OH 1-20 .-Y LJ HN' ^h ' Π Ί; VOH 2-8 and / γ * , N ^F s ' V -Cl NY _ ) , -_-N ! V NH ^F - ^J J oj ' ^F YN ^{1 F} S A- ..-Cl N“ ^J _,.J Г' NH 1 '1 r

- 12 044387

1-18 Π to,. /1 'll ¹ -N N f I ul '>' F r/] ¹ II - OH _lo ' -NI ^th HN f : cr JF Г · ·' OH 1-21 /l,^r^.. O 2-9 and A ^F 5 A --1 nHJ f oj ' J ^{1 Γ} Ύ'ν ^F RAg ^r < _z HO- 1-19 w, ^N / Λ ^Ν , - Cl - Ac CH 1-22 hn-4 v “r' ( 1G 1 Th 2-10 O . - -N i 1 LI ; F ... ,1 1 G II - OH O -N 1N f:: Cl j F : OH

- 13 044387

1- 19-1 U U s, ί. at Mon 1-23 D YN s k L ¹ C1 Ί ' N ^x ^chA b ¹ D 'G he 1-28 w N-, Q S^TV UU 2-2 O '° / C ^g 7 O ^Ζ _ 2-5- 2 D - O di [ 1 '1 1 3-1-1 v N P ABOUT IOH 2-3 Ν - \ V ^ы \ ¹ ji ^a l 1 ³ BUT ^! '^P N - '-Ν V - CI 7L%| BUT ' - 2-6 1у° o..... *AXg N^rr NpW 0 Q- D D NH ^N -N 3-1-2 4° N--. V? ί .--^ ,CI \ ^{1 1 1 N} f Ii X

- 14 044387

2-4 4 .0 fj ογ N -, ( \ /-Г1 < 1 ¹ N - N j X Ί ; 2-6- 1 4° p k/V ^a —N A ¹ _t p 3-2 \\ 0 , N N . Cl cha । HN O ¹ ' 2-5 5 ® //V l >— /.........y - Y ζΥ ² o 2-6- 2 Y N-, R YXY nΑ-γΑ 3-3 4 N - ' N - Г| I ^N ; i 1 Ϊ NI 2-5- 1 0 g Λ A '. . Cl s Ί I N - N ^J .- A ^H JJ o 3-1 π 7.- ¹ -1: ^{11 N} ; Il jCH 3-4 0 _s X^v" 3-5 M N-^ V 3-10 A OH 3-15 N-, A <1 ! । - - And NIU

- 15 044387

- 16 044387

3-21 4-1 \\ 0 N, h Y F'l 'll γ Ί - Ϊ OH 4-6 And s'll 'Νγ ci\ A N V C.I. [ 1 !| υΗ 3-22 A w Cl 4' OH 4-2 'N Cl A. ¹ _b 4-7 -NH Vy ^cl s F 3-23 A N - 4 \i, . ^u 1 Ji J 4-3 w o 'N' . Cl ^nJ J Ί ¹ 4-8 Ύ b N.H. Tskh P I .o 3-24 0 h - N\ ¹ _; F :+ i j 4-4 O +-NP A' .. - . Cl j'Br 4-9 V/'NH • Ί ' ! 1 HE 5-1 Ή G and ¹¹ . ¹ GP 5-6 _ι Ν H A .,, \ J. 1 1 ί ” ; 1 Ί I ΌΗ 6-2 ABOUT <Ν S 4 2 O ^-2 γ 5-2 □ _*NH N b ,1 ^iCl 1 1 IJ 4 5-7 %? OlX ί J nYA/W S HE 7-1 ι Ζ ^{Ν f} \ J Ν-,, Ρ - A ' ^F O Gl 1 J ί i J' OH u, J 1 1' J ί J l·। ^! iOH ' O _ NH V- - _(i |, -L * F ^N -A'GL 7 'Ί / -N 7 N eleven I 1 11 5-3 H ъ _ ⁴ ' 1 1 . Cl G 1 T OH 5-8 L JF y:; ^15Y . WITH . '1111 chi I 7-2 O m- 1 Ί OH -N ^F r% 1 HE '1 1 > J ^F 1 i. -OH

- 17 044387

5-4 l° .....0 Ml V Cl sY J A nAXv4z T IJ OH 5-9 -> ^N Cw ChSH Λ Tu he 7-3 °7. . Cl - ^{0 s} □ i ;' ii N ^J Г- ./ ,· ii ! OH o/ ^K Y । I-ol. ⁰ - ^! , ¹ / ^rJ [. I, TOH 5-5 V n n ^H Vf ^cl A TJJ 1 y T ly OH 6-1 h Ν-^ - Μ ^p and I 1 ^Ν ΐ ri 1 NY 8-1 * 0 < / ^{α F} Ά /=·:' t vrv NJ' OH ^d ' 8-1- 1 l° N —, 1st eluting isomer 8-3- 2 h /-Ν Η0 ^Ζ уХз* 2nd eluting isomer 8-6-2 V Λ xbfxX 0 N n Yr Y / ^;c 2nd eluting isomer

- 18 044387

8-1- 2 ABOUT /=/ /'A 2nd eluting isomer 8-4 ’ r° ,Ν. L' ό ΊΊ U » Ν η 1 1 but J J 9-1 N— _s ' U Cl Cl N ----' .D1 iPr — W N. . J' t 1 N 8-2 ' G) ' G ί g ' N - 1 g 8-5 ' N x ¹ p Γ'ϊ X i ' |I 1 , | HO''X 9-2 /— ⁴ Bi N /— O \ 8-3 ⁴ { Ddd Vn but yb 8-6 l ¹ 0 I N ASL i LLJ O' NN η I 1 ,,0- ¹ '' Γ1 9-3 4 ( ) a ci П z ·ζ NH2 o’ \__ 8-3- 1 N—. bsfip An but ⁷ 1st eluting isomer 8-6- 1 V Ό O^N Νγ> A ” ^: 1st eluting isomer 9-4 V M. Г 1 'N' N j, ^U Ο- ΐ- J · ' ^0H 'Λ λ' 7° ί 1 'N' N r. -A ¹ 1 JI . .-OH ^P 4 '[! ''-IZ!

- 19 044387

9-5 J with K.H. ! 1 h A A ^{0 N} .0' ™ N 9-10 A { ) a ci a g ⁴ _{= 1} = AAA 10-1 o J Ί 1 1. And ^r li I Ϋ ^Г=г 9-6 '1 X 1 N L ci У И ι ί .Y 11 ι |1 ^but >4 ' 9-11 X ( L a ct ' >,- - i. . 10-2 „ 1 r! 'N' ^fU r A i L li n li ' I 4, ' OH _f = _r 9-7- 2 t ΐγΎ 1 η·Χ'Ν'^ ^χ γ|^ ^ΑχΑ hqA Sr 2nd eluting isomer 9-12 φ ,Ν. ¹ AG ι. Г ~ Ό N С 14 ' LJ 10-3 π 1 I N. 1 'N' A , - A ¹ Ν । 1 JL n ^{1 f} Λ LI - 9-7- 1 V А ΓΎΎ а /\'ι XJ ^but Ν 1st eluting isomer 9-13 to 0 ₅ Ηγ' οΖΑΛγκ Λ '7 10-4 A ό N

- 20 044387

9-9 X'N' N has ¹ . , ^П 1 1 L / pi ϊ ! ' G 1 II ' Yr 9-14 VA Yr ¹ an I ' r ^F 1 UNN 1| 1 HG '1 ' J ir J 1 'N' I . .1 1 1 I 1 Ο'Ή h , , J 'HoJ J 1 i 10-5 0 0Η ^α γγ4 ι ADg a γ 10-6 °? l Ν Гк ¹ <>Н [ '1 .1 ' N i L ^F JI lO- ll . 1 N cA f ~ Ϊ ¹ N 1 1 JA Il J, I a 1 11-2- 1 φ n Vy ^ci y A z ^NH Y ¹ 1st eluting isomer mixture 10-7 No. 0 he YY 1 ι AULn Αγ 10- 12 0 1 OH ¹ t 1 'N 1 IDN Hi' 1 LE 'H 11-2- 2 Ό ^Ν iGY ^cl /= ^ν . AAAA νη Y a 2nd eluting isomer mixture 10-8 - : / 10- 13 0 1 _r i L 'N' ri . 1 oH TJ, N 1 1 and and ''/ '' 12 o '1, _f N i IN Cl E I Ί J 1 JN d I 1 - OH ___

- 21 044387

10-9 . And γ ,Ν. Ν ci 1 7 ι ι g Hi .J ^h Ί 1 A ' eleven- 1-1 A N—, °УЪ yyyj An Д .nh vX ^N 1st eluted isomeric mixture 13 Up to 0 a G; II _t -L ⁿ HO ^J . ^J 10- 10 o 1 7 1 N r -Ά * ^J - ⁿ ii ί r 1 1 eleven- 1-2 A Chh uulu YES 2nd eluting isomeric mixture 14 °? 0 ^C| * ¹ II ι ι J II » 1 1 OH, 15 |l o r N i 1 N γη' m ™ ¹ 18-2 \\ 0 VgUD 0 _a 20 But U ri '< 1 I ι Ϊ he 16 i o ri 1 i Ά J. A A ' 1 ι ι ^Hl ' ¹ 18-3 Ν-. A - o ^and L g 21 U 1 A HE 17-1 'UD IL .1 19-1 h° УI” 4 7 'g । ι y 22 W ³ - · U ° A'· 4 1 ¹ ι γ he

- 22 044387

- 23 044387

Table 1(a)

No. approx. Chemical structure No. approx. Chemical structure No. note Chemical structure 1st eluted 2nd eluted A- 39-1 isomer Kommersant GA. 39-2 isomer h _N y _Z -A 40-1 K - Ν s-Ν Tx Oh but ABOUT ; Vn ,Α g I 1st eluting isomer A° ’ G f V N—, • F F y-N A = Ν—. F G 40-2 / , ___ N -N ^s : - TA 41-1 N ' —N —— ^: XI . CH 1 N = 41-2 To XJ.— XI CH 2nd eluted 1st eluted 2nd eluted isomer isomer isomer * O N—. eleven \\ O N—, y-N =- V. Ν—. Cl F U N - 42-1 N—N ' AND HE Q Ί--/ -N—' 42-2 Ν' — Ν - s- N ОН е >— . _Ν =^ 44-1 M. Ν .—' —Ν , OH η 1—' 1st eluted 2nd eluted 1st eluted isomer isomer isomer W N — ы F y ^N “ V N. WITH! F χ-Ν _=< = N. UF Α ^{ν =} 44-2 N P —' AND HE 45-1 Ν „ Ν ---. . Ν 45-2 Ν - Ν - - , Ν 0 — N— M = ν 2nd eluted 1st eluted 2nd eluted isomer isomer isomer Yo A Q A A O A VΝ /=4 A. Ν—. . Cl F _, 46-1 A /n=/ 46-2 A / 49-1 X|' BUT O' > 1st eluted 2nd eluted 1st eluted isomer isomer isomer

- 24 044387

Synthesis of disclosed compounds.

The compounds disclosed herein can be synthesized through a number of specific methods. The examples, which set forth specific synthetic routes, and the general diagrams below are intended to provide guidance to the person of ordinary skill in the art of chemical synthesis to readily understand the solvent, concentration, reagent, protecting group, order of synthetic steps, time, temperature, and the like may be modified as necessary, and the foregoing is fully within the skill and discretion of one of ordinary skill in the art.

Method 1.

Synthesis method 1.

The compound of formula (I) disclosed herein can be synthesized as set forth in Method 1. An appropriate aromatic or heteroaromatic acid is reacted with a halogenating agent in Step 1 to form a halogenated aromatic or heteroaromatic acid. The acid is then reacted with the amidating agent in step 2 to form the amide intermediate. The amide intermediate is then reacted with a sulfonating agent in step 3 to form a thioamide intermediate. Next, the thioamide intermediate is reacted with the oxidizing agent in step 4 with about

- 25 044387 formation of the thiazole ring as shown. The amine group of the thiazole is then converted to a leaving group in step 5 using an activating agent. The leaving group is then replaced with an R ⁴ protected group as shown in step 6. The R ² moiety is then introduced in step 7 by performing a cross-coupling reaction using the appropriate R ² reagent (protected) with the X-halide in the thiazole intermediate. Then, in step 8, the R ⁴ group is deprotected under appropriate conditions depending on the protecting group used, the R ⁴ group is then acylated to introduce an acrylamide moiety as shown, and finally the R ² group is deprotected . Suitable protecting groups and reagents for removing the protecting group are known to those skilled in the art, for example as described in Greene's Protective Groups in Organic Synthesis.

Provided halogenating agents include, but are not limited to, chlorine, bromine, N-chlorosuccinimide and N-bromosuccinimide, optionally performing their function in the presence of a catalyst such as iron or aluminum. It will be readily apparent to those of ordinary skill in the art of chemical synthesis that other halogenating agents and catalysts may be used.

Specified amidating agents include, but are not limited to, N,N'-diisopropylcarbodiimide, N-(3-dimethylaminopyl)-N'-ethylcarbodiimide, benzotriazol-1-yl-oxytripyrrolidinophosphonium hexafluorophosphate, O-(benzotriazol-1-yl)-N,N, N,N'-tetramethyluronium hexafluorophosphate, thionyl chloride, isobutyl chloroformate, diethyl cyanophosphonate, carbonyldiimidazole and polyphosphonic acid anhydride. It will be readily apparent to those of ordinary skill in the art of chemical synthesis that other amidating agents may be used.

Suitable sulfonating agents include, but are not limited to, sulfur, phosphorus pentasulfide, and Lawsson's reagent. It will be readily apparent to those of ordinary skill in the art of chemical synthesis that other sulfonating agents may be used.

Suitable oxidizing agents include, but are not limited to, hydrogen peroxide, iodobenzene diacetate, tert-butyl hydroperoxide, N-bromosuccinimide, and ammonium peroxodisulfate. The average person skilled in the art of chemical synthesis will readily understand that other oxidizing agents can be used.

Provided activators include, but are not limited to, sodium nitrite and tert-butyl nitrite. The person skilled in the art of chemical synthesis will readily appreciate that other activating agents may be used.

Provided cross-coupling reactions include, but are not limited to, Suzuki coupling, Negishi coupling, Hiyama coupling, Kumada coupling, and Stille coupling. One skilled in the art of chemistry will readily appreciate that coupling reactions as shown in Method 1 can be carried out under a number of conditions.

Method 2.

BUT

Stage 1 sulfonating agent combination using R ⁴ (PG) o

means

Stage 2 (PG) R

R ² (PG) - reagent catalyst for cross-coupling reaction

Stage 5

X (1) removal of (acyl) R ⁴ protecting group from R ⁴ y

Stage 3 (PG)

S (2) acylating agent 'n^e^^R ² (PG)

Steps 6 and 7 (1) deprotecting (acyl) R ⁴ group from R ²

--------------S

Stage 8

X

R ²

Synthesis method 2.

Method 2 provides an alternative method for the formation of compounds of formula (I), as disclosed herein. After halogenation in step 1, the R ⁴ -protected group is introduced by reacting with an acid in a coupling reaction in step 2. The oxo group is converted to sulfur using a sulfonating agent in step 3. The thiazole ring is then formed in the presence of an oxidizing agent in step 4. The remaining steps are 5-8 similar to steps 7 and 8 in method 1 described above.

- 26 044387

Method 3.

R ⁴ (PG) ₃ (1) removal of (acyl)K protecting group from R ⁴

s. J J -*

N E X (2) acylating agent

Stage 1 .R ³

R ² (PG)-reagent (acyl) K

S

(1) deprotection of (acyl)E

R groups with R ²

X catalyst for cross-coupling reaction

Stage 2

R ³

R ² (PG) Stage 3 'R ²

Synthesis method 3.

Method 3 provides an alternative method for the formation of compounds of formula (I) as disclosed herein. The R ⁴ group of the isothiazole intermediate is deprotected and acylated in step 1 to introduce an acrylamide moiety. The R ² moiety is then introduced in step 2 by performing a cross-coupling reaction using the appropriate R ² reagent (protected) with the X-halide in the isothiazole intermediate. Finally, the R ² group is deprotected in step 3.

Method 4.

LG

(PG)-reagent

₃ (1) removal of the protecting ( ^a C ^il ) ^group from R ⁴

Stage 1 (2) acylating agent

Stage 2

R ² (PG) - catalyst reagent for cross-coupling reaction Stage 3

S

(1) removal of the protecting group from R ² (PG)

Stage 4

Synthesis method 4.

Method 4 provides an alternative method for the formation of compounds of formula (I), as disclosed herein. After replacing the leaving group in the isothiazole intermediate with a protected R ⁴ group as depicted in Step 1, the R ⁴ group of the intermediate is deprotected and acylated in Step 2 to introduce an acrylamide moiety. The R ² moiety is introduced by performing a cross-coupling reaction in step 3 as in method 1, and the R ² group is deprotected in step 4.

Method 5.

R ³ catalyst for cross-coupling reaction Step 1

R ² (PG) -reagent

R ² (PG) agent

Stage 2

(PG) (PG) -reagent

Stage 3 oxidizer

(PG) activating agent

(PG)

Stage 4

Stage 5

^R4 (PG)s; % .R ³ (1) removal of the protecting (acyl)K ⁴ group from R ⁴ E^R ³

_R2/pr . (2) acylating agent ^S |h e^R ² k sch G) (3) _removal of the protecting group from R ²

Stage 6

Synthesis method 5.

Method 5 provides an alternative method for the formation of compounds of formula (I), as disclosed herein. In this alternative, the R ² moiety is first introduced by performing a cross-coupling reaction with an X-halide in the aromatic or heteroaromatic amide intermediate shown in step 1. The amide intermediate is then reacted with a sulfonating agent in step 2 to form the thioamide intermediate . Oxidation of this intermediate produces an isothiazole ring in step 3. The amine group is then converted to a leaving group in step 4 and then replaced with a protected R ⁴ group in step 5. Finally, in step 6, the R ⁴ group is deprotected and it is reacted with an acylating agent and then the R ² group is deprotected.

- 27 044387

Method 6.

r ⁴ (PG) V Ш metallating agent R ⁴ (PG)

R ³

R ² -reagent

Stage 1

N E Met

R ⁴ (PG) _q (1) removal of the protecting group from R ⁴

R ³ catalyst for cross-coupling reaction Step 2 (2) acylating agent

E R ²

Synthesis method 6.

Method 6 provides an alternative method for the formation of compounds of formula (I), as disclosed herein. In this alternative, the isothiazole intermediate is reacted with a methylating agent to activate the X-halide. The R ² group is then introduced by reacting the activated intermediate with an appropriate R ² reagent (protected). At the last stage, the R ⁴ group is deprotected and it is acylated with the introduction of an acrylamide fragment.

Suitable methylating agents include, but are not limited to, bis(pinacolato)diboron, magnesium, zinc, hexamethyldistannan and n-butyllithium. It will be readily apparent to the person of ordinary skill in the art of chemical synthesis that other methylating agents and catalysts may be used.

Method 7.

R ² (PG) - reagent catalyst for cross-coupling reaction

R ⁴ (PG) reagent

Stage 2

Stage 1

sulfonating

R ² (PG) agent

Stage 3

oxidizing agent (acyl

E. ,R'

Stage 4

(1) removal of the protecting group from R ⁴ (2) acylating agent 'd3) removal of the protecting group from R ²

E R ²

Stage 5

Synthesis method 7.

Method 7 provides an alternative method for the formation of compounds of formula (I) as disclosed herein. The R ² moiety is first introduced by cross-coupling with an X-halide in the aromatic or heteroaromatic acid intermediate shown in Step 1. The acid moiety is then reacted with the appropriate R ⁴ reagent (protected) in the presence of an amidating agent in Step 2.

The carbonyl group of the acid derivative is then converted to a thiocarbonyl group in step 3 using a sulfonating agent. The thioacid derivative is then reacted with an oxidizing agent to form an isothiazole intermediate in step 4. Finally, the R ⁴ group is deprotected and acylated to introduce an acrylamide moiety and the R ² group is deprotected.

Method 8.

- 28 044387

Synthesis method 8.

The compound of formula (II) disclosed herein can be synthesized as set forth in Method 8. An appropriate aromatic or heteroaromatic acid is reacted with an amidating agent in Step 1 to form a primary amide intermediate. The amide is then reacted with an isocyanate-forming reagent and an R ¹⁰ substituted amine to form a urea intermediate. Provided isocyanate-forming agents include oxalyl chloride, thionyl chloride and phosphorus oxychloride. The urea intermediate is then reacted with the cyclizing agent in step 3 to form the quinazolindione ring shown. Provided cyclizing agents include, but are not limited to, bases such as potassium hexamethyldisilazide, potassium tert-butoxide, sodium hydride and phosphazene bases. The R ² moiety is then introduced in step 4 by performing a cross-coupling reaction using the appropriate R ² reagent (protected) with the X-halide in the quinazolindione intermediate. The oxo group of the quinazolindione is then converted to a leaving group in step 5 using an activating agent. Suitable activating agents include, but are not limited to, thionyl chloride, triflate anhydride, phosphorus oxychloride and phosphorus pentachloride. The leaving group is then replaced with an R ⁴ -protected group to form a substituted quinazolinone as shown in step 6. The remaining deprotection-acylation-deprotection sequence shown in steps 7-9 is similar to step 8 in method 1.

Method 9.

activator

R ³ (1) R ⁴ (PG) -reagent

Stage 1

R ² (acyl) AND (PG) - catalyst reagent for cross-coupling reaction Step 3

NFE

Χ ^Ν

O R ¹⁰

R ² (PG) (2) deprotection of R ⁴ (3) acylating agent

Step 2 (1) removal of the protecting group from R ²

Stage 4

R ³

Synthesis method 9.

Method 9 provides an alternative method for the formation of compounds of formula (II) as disclosed herein. The oxo group of the quinazolindione is converted to a leaving group in step 1. Step 2 involves introducing an R ⁴ group (protected), removing the protecting group from the R ⁴ group, and acylating the free R ⁴ group. The R ² group is introduced in step 3 by performing a cross-coupling reaction using the appropriate R ² reagent (protected) with the X-halide in the quinazolindione intermediate. Finally, the R ² group is deprotected.

Method 10.

Synthesis method 10.

The compound of formula (V) disclosed herein can be synthesized as set forth in Method 10. The corresponding anhydride is reacted with hydrazine to form a phthalazinedione ring as shown in Step 1. The R moiety is introduced in Step 2 by performing a cross reaction -coupling using the appropriate R-reagent with the X-halide in the quinazolindione intermediate. The R group is then protected in step 3. The phthalazinedione ring is halogenated twice. The halogenating agent provided includes thionyl chloride, phosphorus oxychloride and oxalyl chloride. One of the groups representing a halogen, then

- 29 044387 is replaced by an R ⁴ protected group to form a substituted phthalazine ring as shown in step 5. Steps 6 and 7 then deprotect the R ⁴ group under appropriate conditions, depending on the protecting group used, and then free R ^{The 4} -group is acylated with the introduction of an acrylamide fragment. At stage 8, R ² is deprotected. Finally, the R ¹⁰ moiety is introduced in step 9 by performing a cross-coupling reaction using the appropriate R ¹⁰ reagent with the X-halide in the phthalazine intermediate.

Method 11.

activating agent benzotriazole

Stage 1

R ² -reagent, catalyst for cross-coupling reaction Stage 3 (PG)

-------------► N (2) acylating agent km O R ¹⁰

R ² (1) removal ₄ . ^r3 protecting group with (acyl)I E ⁼ \ R ⁴ NA / A ²

Stages 4 and 5

Synthesis method 11.

Method 11 provides an alternative method for the formation of compounds of formula (II) as disclosed herein. The oxo group of the quinazolindione is converted to a leaving group in step 1. The R ⁴ group (protected) is introduced in step 2. The R ² group is introduced in step 3 by performing a cross-coupling reaction using the appropriate R ² reagent (protected) with the X-halide in the quinazolindione intermediate. Finally, the R ⁴ group is deprotected and then acylated in steps 4 and 5.

Pharmaceutical compositions, dosage and routes of administration.

Also provided herein are pharmaceutical compositions that contain a compound disclosed herein together with a pharmaceutically acceptable excipient, such as, for example, a diluent or carrier. Compounds and pharmaceutical compositions suitable for use in the present invention are those where the compound can be administered in an effective amount to achieve its intended purpose. The administration of the compound is described in more detail below.

Suitable pharmaceutical formulations can be determined by one skilled in the art depending on the route of administration and the required dose. See, for example, Remington's Pharmaceutical Sciences, 1435-712 ( ^18th ed., Mack Publishing Co, Easton, PA, 1990). The formulations may affect the physical state, stability, in vivo release rate, and in vivo elimination rate of the administered agents. Depending on the route of administration, the appropriate dose may be calculated according to body weight, body surface area or organ size. It is customary for those skilled in the art to further refine the calculations necessary to determine the appropriate treatment dose without undue experimentation, especially in light of the dosage information and assays disclosed herein, as well as the pharmacokinetic data that can be obtained from clinical trials in animals or people.

The phrases pharmaceutically acceptable or pharmacologically acceptable refer to molecular substances and compositions that do not cause adverse, allergic or other adverse reactions when administered to an animal or human. As used herein, the term pharmaceutically acceptable includes all possible solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonic agents and absorption retarding agents, and the like. The use of such excipients for pharmaceutically active substances is widely known in the art. Unless any conventional media or agent is incompatible with the therapeutic compositions, their use in the therapeutic compositions is intended. Additional active ingredients may also be included in the compositions. In illustrative embodiments, the formulation may contain corn syrup powder, high oleic safflower oil, coconut oil, soybean oil, L-leucine, tribasic calcium phosphate, L-tyrosine, L-proline, L-lysine acetate, DATEM (emulsifier), L-glutamine , L-valine, dibasic potassium phosphate, L-isoleucine, L-arginine, L-alanine, glycine, L-asparagine monohydrate, L-serine, potassium citrate, L-threonine, sodium citrate, magnesium chloride, L-histidine, L -methionine, ascorbic acid, calcium carbonate, L-glutamic acid, L-cystine dihydrochloride, L-tryptophan, L-aspartic acid, choline chloride, taurine, m-inositol, ferrous sulfate, ascorbyl palmitate, zinc sulfate, L-carnitine, alpha-tocopheryl acetate, sodium chloride, niacinamide, mixed tocopherols, calcium pantothenate, copper sulfate, thiamine chloride hydrochloride, vitamin A palmitate, manganese sulfate, riboflavin, pyridoxine hydrochloride, folic acid, beta-carotene, potassium iodide, phylloquinone, biotin, sodium selenate, chromium trichloride, sodium molybdate, vitamin D3 and cyanocobalamin.

The compound may be present in the pharmaceutical composition as a pharmaceutically acceptable salt. As used herein, the phrase pharmaceutically acceptable salts includes, for example, base addition salts and acid addition salts.

Pharmaceutically acceptable base addition salts can be formed with metals or amines, such as alkali and alkaline earth metals or organic amines. Pharmaceutically acceptable salts of the compounds can also be prepared using a pharmaceutically acceptable cation. Suitable pharmaceutically acceptable cations are widely known to those skilled in the art and include alkali, alkaline earth, ammonium and quaternary ammonium cations. It is also possible to use carbonates or bicarbonates. Examples of metals used as cations are sodium, potassium, magnesium, ammonium, calcium or ferric iron and the like. Examples of suitable amines include isopropylamine, trimethylamine, histidine, N,N'-dibenzylethylenediamine, chloroprocaine, choline, diethanolamine, dicyclohexylamine, ethylenediamine, N-methylglucamine and procaine.

Pharmaceutically acceptable acid addition salts include salts of inorganic or organic acids. Examples of suitable acid addition salts include hydrochlorides, formates, acetates, citrates, salicylates, nitrates, phosphates. Other suitable pharmaceutically acceptable salts are well known to those skilled in the art and include, for example, those formed with formic, acetic, citric, oxalic, tartaric or mandelic acids, with hydrochloric acid, hydrobromic acid, sulfuric acid or phosphoric acid; with organic carboxylic, sulfonic, sulfo- or phosphoric acids or N-substituted sulfamic acids, for example acetic acid, trifluoroacetic acid (TFA), propionic acid, glycolic acid, succinic acid, maleic acid, hydroxymaleic acid, methylmaleic acid, fumaric acid, malic acid acid, tartaric acid, lactic acid, oxalic acid, gluconic acid, glucaric acid, glucuronic acid, citric acid, benzoic acid, cinnamic acid, mandelic acid, salicylic acid, 4-aminosalicylic acid, 2-phenoxybenzoic acid, 2-acetoxybenzoic acid, embonic acid, nicotinic acid or isonicotinic acid, and with amino acids such as the 20 alpha amino acids involved in protein synthesis in nature, such as glutamic acid or aspartic acid, and with phenylacetic acid, methanesulfonic acid, ethanesulfonic acid, 2- hydroxyethanesulfonic acid, ethane-1,2-disulfonic acid, benzenesulfonic acid, 4-methylbenzenesulfonic acid, naphthalene-2-sulfonic acid, naphthalene-1,5-disulfonic acid, 2- or 3-phosphoglycerate, glucose-6-phosphate, N -cyclohexylsulfamic acid (to form cyclamates) or with other acidic organic compounds such as ascorbic acid.

Pharmaceutical compositions containing the compounds disclosed herein can be prepared in a conventional manner, such as through conventional mixing, dissolving, granulating, dragee-making, trituration, emulsification, encapsulation, entrapment, or lyophilization processes. The appropriate formulation depends on the route of administration chosen.

Suitable compositions for oral administration can be readily formulated by combining the compound disclosed herein with pharmaceutically acceptable excipients, such as carriers well known in the art. Such excipients and carriers allow the compounds of the present invention to be formulated into tablets, pills, dragees, capsules, liquids, gels, syrups, slurries, suspensions, etc. for oral administration by the patient to be treated. Pharmaceutical preparations for oral use can be prepared by adding a solid excipient to a compound disclosed herein, optionally grinding the resulting mixture, and processing the mixture into granules after adding suitable excipients, if necessary, to obtain tablets or dragee cores. Suitable excipients include, for example, fillers and cellulosic preparations. If necessary, substances can be added to improve the disintegration of tablets. Pharmaceutically acceptable ingredients are well known for various types of formulations and may include, for example, binders (e.g., natural or synthetic polymers), lubricants, surfactants, sweeteners and flavoring agents, coating materials, preservatives, colorants, thickeners, excipients, antimicrobial agents, antioxidants and carriers for various types of formulations.

If a therapeutically effective amount of a compound disclosed herein is administered orally, the composition will typically be in the form of a solid formulation (e.g., tablet, capsule, pellet, powder or lozenge) or liquid formulation (e.g., aqueous suspension, solution, tincture or syrup).

When administered in tablet form, the composition may further comprise a functional solid and/or a solid carrier such as gelatin or an excipient. The tablet, capsule and powder may contain from about 1 to about 95% compound and preferably from about 15 to about 90% compound.

When administered in the form of a liquid or suspension, a functional liquid and/or a liquid carrier such as water, a hydrocarbon carrier, or oils of animal or plant origin may be added. The liquid form of the composition may further contain saline, solutions of sugar alcohols, solutions of dextrose or other saccharides or glycols. When administered in liquid or suspension form, the composition may contain from about 0.5 to about 90% by weight of a compound disclosed herein, and preferably from about 1 to about 50% by weight of a compound disclosed herein. In one embodiment, the liquid carrier is provided to be non-aqueous or substantially non-aqueous. The composition, intended for administration in liquid form, may be supplied as a rapidly dissolving solid formulation or as a suspension for dissolution immediately prior to administration.

When a therapeutically effective amount of a compound disclosed herein is administered by intravenous, dermal, or subcutaneous injection, the composition is in the form of a pyrogen-free, parenterally acceptable aqueous solution. The preparation of such parenterally acceptable solutions having appropriate pH, isotonicity, stability, etc. is within the skill of one skilled in the art. A preferred composition for intravenous, dermal, or subcutaneous injection typically contains, in addition to the compound disclosed herein, an isotonic carrier medium. Such compositions can be prepared for administration as solutions of the free base or pharmacologically acceptable salts in water suitably mixed with a surfactant such as hydroxypropylcellulose. Dispersions can also be obtained in glycerin, liquid polyethylene glycols and mixtures thereof, as well as in oils. Under normal conditions of storage and use, such preparations may not necessarily contain a preservative to prevent the growth of microorganisms.

Injectable compositions may include sterile aqueous solutions, suspensions or dispersions and sterile powders for the immediate preparation of sterile injectable solutions, suspensions or dispersions. In all embodiments, the form must be sterile and must be fluid to the extent that it can be easily syringed. It must be stable under the conditions of manufacture and storage and must resist the contaminating action of microorganisms such as bacteria and fungi through the optional inclusion of a preservative. The carrier may be a solvent or dispersion medium containing, for example, water, ethanol, a polyol (eg, glycerin, propylene glycol and liquid polyethylene glycol, etc.), suitable mixtures thereof, and vegetable oils. In one embodiment, the carrier is provided to be non-aqueous or substantially non-aqueous. Proper fluidity can be maintained, for example, by the use of a coating such as lecithin, by maintaining the desired particle size of the compound in the dispersion embodiment, and by the use of surfactants. Prevention of exposure to microorganisms can be achieved by various antibacterial and antifungal agents, such as parabens, chlorobutanol, phenol, sorbic acid, thiomersal, etc. In many embodiments, it will be preferred to include isotonic agents, such as sugars or sodium chloride. Prolonged absorption of injectable compositions can be achieved by using absorption retarding agents in the compositions, such as aluminum monostearate and gelatin.

Sterile injection solutions are prepared by supplementing the active compounds in the required quantity in an appropriate diluent as needed with the various other ingredients listed above, followed by sterilization by filtration. Typically, dispersions are prepared by incorporating various sterilized active ingredients into a sterile carrier medium that contains the basic dispersion medium and the required other ingredients listed above. In the embodiment of sterile powders intended for the preparation of sterile injectable solutions, preferred production methods are vacuum drying and freeze drying techniques that provide a powder of the active ingredient plus any additional required ingredient from a pre-sterile filtered solution thereof.

Slow-release or sustained-release formulations may also be prepared to achieve controlled release of the active compound in contact with body fluids in the gastrointestinal tract and to provide a substantially constant and effective plasma level of the active compound. For example, release may be controlled by one or more of dissolution, diffusion, and ion exchange. Additionally, a slow-release approach may enhance absorption through saturable or limiting pathways in the gastrointestinal tract. For example, for this purpose, the compound may be incorporated into a polymer matrix of a biodegradable polymer, a water-soluble polymer, or a mixture of both and optionally suitable surfactants. In this context, embedding may mean the incorporation of microparticles into a matrix of polymers. Controlled release formulations are also prepared by encapsulating dispersed microparticles or emulsified microdroplets from

- 32 044387 with the power of well-known dispersion or emulsion coating technologies.

For administration by inhalation, the compounds of the present invention are conveniently delivered in the form of a nebulized aerosol from pressurized packages or by nebulizer using a suitable propellant. In a pressurized aerosol embodiment, a dosage unit may be defined by providing a valve to deliver a metered amount. For use in an inhaler or insufflator, capsules and cartridges, for example gelatin, can be formulated containing a powder mixture of the compound and a suitable powder base such as lactose or starch.

The compounds disclosed herein may be formulated for parenteral administration by injection (eg, bolus injection or continuous infusion). Injectable formulations may be presented in unit dosage form (eg, ampoules or multi-dose containers) with an added preservative. The compositions may take forms such as suspensions, solutions or emulsions in oily or aqueous carrier vehicles, and may contain formulation auxiliaries such as suspending, stabilizing and/or dispersing agents.

Pharmaceutical compositions intended for parenteral administration include aqueous solutions of the compounds in water-soluble form. In addition, suspensions of the compounds can be prepared as appropriate oily injection suspensions. Suitable lipophilic solvents or carrier media include fatty oils or synthetic fatty acid esters. Aqueous injection suspensions may contain substances that increase the viscosity of the suspension. Optionally, the suspension may also contain suitable stabilizers or agents that increase the solubility of the compounds and provide highly concentrated solutions. Alternatively, the composition of the present invention may be in powder form for reconstitution with a suitable carrier medium (eg, sterile pyrogen-free water) prior to use.

The compounds disclosed herein may also be formulated into compositions for rectal administration, such as suppositories or retention enemas (eg, containing traditional suppository bases). In addition to the formulations described previously, the compounds may also be formulated as a depot formulation. Such long-acting formulations can be administered by implantation (eg, subcutaneously or intramuscularly) or by intramuscular injection. Thus, for example, the compounds may be formulated using suitable polymeric or hydrophobic materials (eg, as an emulsion in a suitable oil) or ion exchange resins, or as sparingly soluble derivatives, eg, as a sparingly soluble salt.

In particular, the compound disclosed herein can be administered orally, buccally or sublingually in the form of tablets containing excipients such as starch or lactose, or in capsules or vaginal suppositories, either alone or mixed with excipients, or in in the form of tinctures or suspensions containing flavoring or coloring agents. Such liquid preparations can be prepared using pharmaceutically acceptable additives such as suspending agents. The compound can also be administered parenterally, for example intravenously, intramuscularly, subcutaneously or intracoronarily. For parenteral administration, the compound is best administered in the form of a sterile aqueous solution, which may contain other substances, such as salts or sugar alcohols such as mannitol or glucose, to render the solution isotonic with blood.

For veterinary use, the compound disclosed herein is administered in a suitable acceptable formulation in accordance with normal veterinary practice. The veterinarian can easily determine the dosage regimen and route of administration that is most appropriate for a particular animal.

In some embodiments, all of the necessary components for treating a KRAS disorder using a compound disclosed herein, either alone or in combination with another agent, or for an intervention traditionally performed to treat such disorder, may be packaged in a kit . Specifically, the present invention provides a kit for use in a therapeutic intervention for a disease, containing a packaged set of drugs that include a compound disclosed herein, as well as buffers and other components intended to obtain the delivered forms of these drugs, and/or devices for the delivery of such drugs, and/or any agents that are used in combination therapy with a compound disclosed herein and/or instructions for the treatment of a disease contained in the packaging of the drugs. The instructions may be recorded on any tangible medium, such as printed paper or computer-readable magnetic or optical media, or the instructions may be provided in the form of a link to a remote computer data source, such as a World Wide Web page accessible via the Internet.

A therapeutically effective amount means an amount effective to treat or

- 33 044387 to prevent the development or to alleviate existing symptoms in the subject being treated. Determination of effective amounts is within the skill of the art, especially in light of the detailed disclosure presented herein. Generally, a therapeutically effective dose refers to that amount of a compound that produces the desired effect. For example, in one preferred embodiment, a therapeutically effective amount of a compound disclosed herein reduces KRAS activity by at least 5% compared to control, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least at least 65%, at least 70%, at least 75%, at least 80%, at least 85% or at least 90%.

The amount of compound administered may depend on the subject being treated, the age, health status, sex and weight of the subject, the type of concurrent treatment (if any), the severity of the disease, the nature of the desired effect, the method and frequency of treatment, and the judgment of the attending physician. Dosing frequency may also depend on pharmacodynamic effects on arterial oxygen pressure. However, the most preferred dosage may be tailored to the individual subject as will be understood and can be determined by one skilled in the art without undue experimentation. This usually involves adjusting the standard dose (eg, reducing the dose if the patient has low body weight).

Although individual needs vary, determination of optimal ranges of effective amounts of a compound is within the skill of those skilled in the art. For administration to humans for the definitive or prophylactic treatment of conditions and disorders identified herein, for example, typical doses of the compounds of the present invention may be from about 0.05 to about 50 mg/kg/day, such as at least 0.05 mg/day kg, at least 0.08 mg/kg, at least 0.1 mg/kg, at least 0.2 mg/kg, at least 0.3 mg/kg, at least 0.4 mg/ kg or at least 0.5 mg/kg and preferably 50 mg/kg or less, 40 mg/kg or less, 30 mg/kg or less, 20 mg/kg or less or 10 mg/kg or less, which may range, for example, from about 2.5 mg/day (0.5 mg/kg x 5 kg) to about 5000 mg/day (50 mg/kg x 100 kg). For example, dosages of the compounds may range from about 0.1 to about 50 mg/kg/day, from about 0.05 to about 10 mg/kg/day, from about 0.05 to about 5 mg/kg/day, from about 0.05 to about 3 mg/kg/day, about 0.07 to about 3 mg/kg/day, about 0.09 to about 3 mg/kg/day, about 0.05 to about 0.1 mg/kg/day, about 0.1 to about 1 mg/kg/day, about 1 to about 10 mg/kg/day, about 1 to about 5 mg/kg/day, about 1 to about 3 mg/kg/day, about 3 to about 500 mg/day, about 5 to about 250 mg/day, about 10 to about 100 mg/day, about 3 to about 10 mg/day, or about 100 to approximately 250 mg/day. Such doses may be administered in a single dose or they may be divided into several doses.

Methods of using G12C KRAS inhibitors.

The present invention provides a method of inhibiting RAS-mediated cell signaling, comprising contacting the cell with an effective amount of one or more compounds disclosed herein. Inhibition of RAS-mediated signal transduction can be assessed and demonstrated by a wide range of pathways known in the art. Non-limiting examples include demonstrating (a) decreased GTPase activity of RAS; (b) decreasing the binding affinity of GTP or increasing the binding affinity of GDP; (c) increasing Koff GTP or decreasing Koff GDP; (d) reducing the levels of signal transduction molecules located downstream in the RAS pathway, for example reducing the levels of pMEK, pERK or pAKT; and/or (f) reducing the extent of binding of the RAS complex to downstream signaling molecules, including but not limited to Raf. Kits and commercially available assays can be used to determine one or more of the above.

The disclosure also provides methods of using the compounds or pharmaceutical compositions of the present invention to treat disease states, including, without limitation, conditions associated with the G12C KRAS, HRAS, or NRAS mutation (eg, cancer).

In some embodiments, a method of treating cancer is provided, the method comprising administering an effective amount of any of the above pharmaceutical compositions containing a compound disclosed herein to a subject in need thereof. In some embodiments, the cancer is mediated by the G12C mutation of KRAS, HRAS, or NRAS. In various embodiments, the cancer is pancreatic cancer, colorectal cancer, or lung cancer. In some embodiments, the cancer is gallbladder cancer, cancer

- 34 044387 thyroid gland and bile duct cancer.

In some embodiments, the present invention provides a method of treating a disorder in a subject in need thereof, the method comprising determining whether the subject has a G12C KRAS, HRAS, or NRAS mutation, and if the subject is determined to have a G12C KRAS, HRAS, or NRAS, then, is administering to a subject a therapeutically effective dose of at least one compound disclosed herein, or a pharmaceutically acceptable salt thereof.

The disclosed compounds inhibit anchorage protein-independent cell growth and therefore have the potential to inhibit tumor metastasis. Accordingly, in another embodiment, the present invention provides a method of inhibiting tumor metastases, the method comprising administering an effective amount of a compound disclosed herein.

G12C KRAS, HRAS, or NRAS mutations have also been identified in hematologic malignancies (eg, cancers that involve the blood, bone marrow, and/or lymph nodes). Accordingly, some embodiments are directed to administering the disclosed compounds (eg, in the form of a pharmaceutical composition) to a patient in need of treatment for hematological malignancies. Such forms of cancer include, but are not limited to, leukemias and lymphomas. For example, the compounds disclosed herein can be used to treat diseases such as acute lymphoblastic leukemia (ALL), acute myelogenous leukemia (AML), chronic lymphocytic leukemia (CLL), small lymphocyte lymphoma (SLL), chronic myelogenous leukemia (CML), acute monocytic leukemia (AMoL) and/or other types of leukemia. In other embodiments, the compounds are useful for treating lymphomas, such as all subtypes of Hodgkin's lymphoma or non-Hodgkin's lymphomas. In various embodiments, the compounds are useful for treating plasma cell malignancies such as multiple myeloma, mantle cell lymphoma, and Waldenström's macroglobulinemia.

Determining whether a tumor or cancer contains a G12C KRAS, HRAS, or NRAS mutation can be made by assessing the nucleotide sequence encoding a KRAS, HRAS, or NRAS protein, by assessing the amino acid sequence of a KRAS, HRAS, or NRAS protein, or by assessing the characteristics of a putative mutant KRAS protein , HRAS or NRAS. The sequence of wild type human KRAS, HRAS or NRAS is known in the art (eg accession number NP203524).

Methods for detecting a mutation in a KRAS, HRAS, or NRAS nucleotide sequence are known to those skilled in the art. Such methods include, but are not limited to, polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) assays, polymerase chain reaction-single-strand conformational polymorphism (PCR-SSCP) assays, PCR-based assays real-time, sequencing of PCR products, assays using mutant allele specific amplification (MASA), direct sequencing, primer extension reactions, electrophoresis, ligation of oligonucleotide probes, hybridization assays, TaqMan assays, SNP genotyping, assays high-resolution melting and microarray analyses. In some embodiments, samples are assessed for G12C KRAS, HRAS, or NRAS mutations using real-time PCR. Real-time PCR uses fluorescent probes specific for the G12C KRAS, HRAS, or NRAS mutations. When a mutation is present, the probe binds and fluorescence is detected. In some embodiments, the G12C mutation of KRAS, HRAS, or NRAS is identified using a method of direct sequencing of specific regions (eg, exon 2 and/or exon 3) in the KRAS, HRAS, or NRAS gene. This technique will allow us to identify all possible mutations in the sequenced region.

Methods for detecting mutations in a KRAS, HRAS, or NRAS protein are known to those skilled in the art. Such methods include, but are not limited to, detection of mutant KRAS, HRAS or NRAS using a binding agent (eg, antibody) specific for the mutant protein, protein electrophoresis and Western blotting, and direct peptide sequencing.

Methods for determining whether a tumor or cancer contains a G12C KRAS, HRAS, or NRAS mutation may use a number of samples. In some embodiments, the sample is collected from a subject having a tumor or cancer. In some embodiments, the sample is a fresh tumor/cancer sample. In some embodiments, the sample is a frozen tumor/cancer sample. In some embodiments, the sample is a formalin-fixed, paraffin-embedded sample. In some embodiments, the sample is a circulating tumor cell (CTC) sample. In some embodiments, the sample is processed into a cell lysate. In some embodiments, the sample is processed into DNA or RNA.

The present invention also relates to a method of treating a hyperproliferative disorder in a mammal, which comprises administering to said mammal a therapeutically effective

- 35044387 effective amount of a compound disclosed herein, or a pharmaceutically acceptable salt thereof. In some embodiments, the method relates to treating a subject who is suffering from cancer, such as acute myeloid leukemia, adolescent cancer, childhood adrenal cancer, AIDS-related cancers (eg, lymphoma and Kaposi's sarcoma), anal cancer , appendix cancer, astrocytomas, atypical teratoid, basal cell carcinoma, bile duct cancer, bladder cancer, bone cancer, brainstem glioma, brain tumor, breast cancer, bronchial tumors, Burkitt's lymphoma, carcinoid tumor, atypical teratoid , embryonal tumors, embryonal cell tumor, primary lymphoma, cervical cancer, childhood cancers, chordoma, cardiac tumors, chronic lymphocytic leukemia (CLL), chronic myelogenous leukemia (CML), chronic myeloproliferative disorders, colon cancer, colorectal cancer , craniopharyngioma, cutaneous T-cell lymphoma, extrahepatic ductal carcinoma in situ (DCIS), embryonal tumors, CNS cancer, endometrial cancer, ependymoma, esophageal cancer, esthesioneuroblastoma, Ewing's sarcoma, extracranial germinal cell tumor, extragonadal germinal cell tumor, cancer eyes, fibrous bone histiocytoma, gallbladder cancer, GI cancer, GI carcinoid tumor, gastrointestinal stromal tumors (GIST), germinal cell tumor, gestational trophoblastic tumor, hairy cell leukemia, head and neck cancer, heart cancer, liver cancer, lymphoma Hodgkin's, hypopharyngeal cancer, intraocular melanoma, pancreatic islet tumors, pancreatic neuroendocrine tumors, kidney cancer, laryngeal cancer, lip and oral cavity cancer, liver cancer, lobular carcinoma in situ (LCIS), lung cancer, lymphoma, metastatic squamous cell carcinoma of the neck with occult primary disease, midline carcinoma, oral cancer, multiple endocrine neoplasia syndromes, multiple myeloma/plasma cell neoplasia, mycosis fungoides, myelodysplastic syndromes, myelodysplastic/myeloproliferative neoplasias, multiple myeloma, Merkel cell carcinoma, malignant mesothelioma, malignant fi bronze histiocytoma of bone and osteosarcoma, nasal and sinus cancer, nasopharyngeal cancer, neuroblastoma, non-Hodgkin's lymphoma, non-small cell lung cancer (NSCLC), oral cavity cancer, lip and oral cavity cancer, oropharyngeal cancer, ovarian cancer, pancreatic cancer, papillomatosis, paraganglioma, cancer paranasal sinuses and nasal cavity, parathyroid cancer, penile cancer, pharyngeal cancer, pleuropulmonary blastoma, primary central nervous system lymphoma (CNS, prostate cancer, rectal cancer, transitional cell cancer, retinoblastoma, rhabdomyosarcoma, salivary gland cancer, skin cancer , gastric cancer (GIT), small cell lung cancer, small intestinal cancer, soft tissue sarcoma, T-cell lymphoma, testicular cancer, throat cancer, thymoma and thymus cancer, thyroid cancer, transitional cell cancer of the renal pelvis and ureter, trophoblastic tumor , unusual childhood cancers, urethral cancer, uterine sarcoma, vaginal cancer, vulvar cancer or virus-induced cancer. In some embodiments, the method relates to the treatment of a noncancerous hyperproliferative disorder, such as benign cutaneous hyperplasia (eg, psoriasis), restenosis, or a prostate-related disorder (eg, benign prostatic hypertrophy (BPH)).

In some embodiments, the treatment methods are directed to the treatment of lung cancers, the methods comprising administering an effective amount of any of the above-described compounds (or a pharmaceutical composition containing the same) to a subject in need thereof. In some embodiments, the lung cancer is non-small cell lung cancer (NSCLC), such as adenocarcinoma, squamous cell lung cancer, or large cell lung cancer. In some embodiments, the lung cancer is small cell lung cancer. Other types of lung cancer that can be treated with the disclosed compounds include, but are not limited to, glandular tumors, carcinoid tumors and undifferentiated cancers.

The present invention further provides methods for modulating the activity of a G12C mutant KRAS, HRAS or NRAS protein by contacting the protein with an effective amount of a compound of the present invention. Modulation may be inhibition or activation of protein activity. In some embodiments, the present invention provides methods for inhibiting protein activity by contacting a G12C mutant KRAS, HRAS, or NRAS protein with an effective amount of a compound of the present invention in solution. In some embodiments, the present invention provides methods for inhibiting the activity of a G12C mutant KRAS, HRAS, or NRAS protein by bringing a cell, tissue, or organ that expresses the protein of interest into contact. In some embodiments, the present invention provides methods for inhibiting protein activity in a subject, including, but not limited to, rodents and mammals (eg, humans), by administering to the subject an effective amount of a compound of the present invention. In some embodiments, the modulation percentage is greater than 25, 30, 40, 50, 60, 70, 80, or 90%. In some embodiments, the percentage of inhibition is greater than 25, 30, 40, 50, 60, 70, 80, or 90%.

In some embodiments, the present invention provides methods for inhibiting

- 36 044387 inhibiting G12C KRAS, HRAS or NRAS activity in a cell by contacting said cell with an amount of a compound of the present invention sufficient to inhibit G12C KRAS, HRAS or NRAS activity in said cell. In some embodiments, the present invention provides methods for inhibiting G12C KRAS, HRAS, or NRAS activity in tissue by contacting said tissue with an amount of a compound of the present invention sufficient to inhibit G12C KRAS, HRAS, or NRAS activity in said tissue. In some embodiments, the present invention provides methods for inhibiting G12C KRAS, HRAS, or NRAS activity in an organism by contacting the organism with an amount of a compound of the present invention sufficient to inhibit G12C KRAS, HRAS, or NRAS activity in the organism. In some embodiments, the present invention provides methods for inhibiting G12C KRAS, HRAS, or NRAS activity in an animal by contacting said animal with an amount of a compound of the present invention sufficient to inhibit G12C KRAS, HRAS, or NRAS activity in said animal. In some embodiments, the present invention provides methods of inhibiting G12C KRAS, HRAS, or NRAS activity in a mammal by contacting said mammal with an amount of a compound of the present invention sufficient to inhibit G12C KRAS, HRAS, or NRAS activity in said mammal. In some embodiments, the present invention provides methods of inhibiting G12C KRAS, HRAS, or NRAS activity in a human by contacting said human with an amount of a compound of the present invention sufficient to inhibit G12C KRAS, HRAS, or NRAS activity in said human. The present invention provides methods for treating a disease mediated by G12C KRAS, HRAS or NRAS activity in a subject in need of such treatment.

Combination therapy.

The present invention also provides methods for implementing combination therapy options in which an agent known to modulate other pathways, or other components of the same pathway, or even overlapping sets of target enzymes, is used in combination with a compound of the present invention or a pharmaceutically acceptable formulation thereof. salt. In one aspect, such therapy includes, without limitation, the combination of one or more compounds of the present invention with chemotherapeutic agents, therapeutic antibodies, and radiation therapy designed to provide a synergistic or additive therapeutic effect.

A variety of chemotherapeutic agents are currently known in the art and can be used in combination with the compounds of the present invention. In some embodiments, the chemotherapeutic agent is selected from the group consisting of mitotic inhibitors, alkylating agents, antimetabolites, intercalating antibiotics, growth factor inhibitors, cell cycle inhibitors, enzymes, topoisomerase inhibitors, biological response modifiers, antihormones, angiogenesis inhibitors, and antiandrogens. Non-limiting examples include chemotherapeutic agents, cytotoxic agents and non-peptide small molecules such as Gleevec® (imatinib mesylate), Kyprolis® (carfilzomib), Velcade® (bortezomib), Casodex (bicalutamide), Iressa® (gefitinib) and Adriamycin, as well as carrier chemotherapeutic agents. Non-limiting examples of chemotherapeutic agents include alkylating agents such as thiotepa and cyclophosphamide (CYTOXANTM); alkylsulfonates such as busulfan, improsulfan and piposulfan; aziridines such as benzodopa, carboquone, meturedopa and uredopa; ethylene imines and methyl melamines, including altretamine, triethylene melamine, triethylene phosphoramide, triethylene thiophosphoramide and trimethylol melamine; nitrogen mustards, such as chlorambucil, chlornaphasine, chlorphosphamide, estramustine, ifosfamide, mechlorethamine, mechlorethamine oxide hydrochloride, melphalan, novembiquin, phenesterine, prednimustine, trofosfamide, uracil mustard; nitrosoureas such as carmustine, chlorzotocin, fotemustine, lomustine, nimustine, ranimustine; antibiotics such as aclacinomycins, actinomycin, anthramycin, azaserin, bleomycins, cactinomycin, calicheycin, carabicin, carminomycin, carcinophilin, CasodexTM, chromomycins, dactinomycin, daunorubicin, detorubicin, 6-diazo-5-oxo-norleucine, doxorubicin, epirubicin, esorubicin qing, idarubicin, marcellomycin, mitomycins, mycophenolic acid, nogalamycin, olivomycins, peplomycin, potfiromycin, puromycin, quelamycin, rhodorubicin, streptonigrin, streptozocin, tubercidin, ubenimex, cinostatin, zorubicin; antimetabolites such as methotrexate and 5-fluorouracil (5-FU); folic acid analogs such as denopterin, methotrexate, pteropterin, trimetrexate; purine analogs such as fludarabine, 6-mercaptopurine, thiamiprin, thioguanine; pyrimidine analogues such as ancytabine, azacitidine, 6-azauridine, carmofur, cytarabine, dideoxyuridine, doxifluridine, enocytabine, floxuridine; androgens such as calusterone, dromostanolone propionate, epithiostanol, mepithiostane, testolactone; inhibitors of adrenal hormone synthesis, such as aminoglutethimide, mitotane, trilostane; com

- 37 044387 folic acid pensators, such as frolinic acid; aceglatone; aldophosphamide glycoside; aminolevulinic acid; amsacrine; bestrabucil; bisantrene; edatraxate; defofamine; demecolcine; diaziquon; elfomitin; elliptinium acetate; ethoglucide; gallium nitrate; hydroxyurea; lentinan; lonidamine; mitoguazone; mitoxantrone; mopidamole; nitracrine; pentostatin; phenomet; pirarubicin; podophyllic acid; 2-ethylhydrazide; procarbazine; PSK; razoxane; sisofiran; spirogermanium; tenuazonic acid; triaziquon; 2,2',2-trichlorotriethylamine; urethane; vindesine; dacarbazine; mannomustin; mitobronitol; mitolactol; pipobromance; gacytosine; arabinoside (Ara-C); cyclophosphamide; thiotepa; taxanes, such as paclitaxel and docetaxel; retinoic acid; esperamycins; capecitabine, and pharmaceutically acceptable salts, acids or derivatives of any of the above.

Also included as suitable cell-enhancing chemotherapeutic agents are antihormonal agents that act to regulate or inhibit the action of hormones on tumors, such as antiestrogens, including, for example, tamoxifen, (Nolvadex™), raloxifene, aromatase inhibitors 4(5 )-imidazoles, 4-hydroxytamoxifen, trioxifene, keoxifene, LY 117018, onapristone and toremifene (Fareston); as well as antiandrogens such as flutamide, nilutamide, bicalutamide, leuprolide and goserelin; chlorambucil; gemcitabine; 6-thioguanine; mercaptopurine; methotrexate; platinum analogues such as cisplatin and carboplatin; vinblastine; platinum; etoposide (VP-16); ifosfamide; mitomycin C; mitoxantrone; vincristine; vinorelbine; navelbine; Novantrone; teniposide; daunomycin; aminopterin; xeloda; ibandronate; camptothecin-11 (CPT-11); topoisomerase inhibitor RFS 2000; difluoromethylornithine (DMFO).

If necessary, the compounds or pharmaceutical composition of the present invention can be used in combination with commonly prescribed anti-cancer drugs such as Herceptin®, Avastin®, Erbitux®, Rituxan®, Taxol®, Arimidex®, Taxotere®, ABVD, Avicin, Abagovomab, Acridine carboxamide, adecatumumab, 17-N-allylamino-17-demethoxygeldanamycin, alpharadine, alvocidib, 3-aminopyridine-2-carboxaldehyde thiosemicarbazone, amonafide, anthracenedione, CD22 immunotoxins, antitumor, antitumorigenic herbs, apaziquone, atiprimod, azathioprine, white tecan, bendamustine, BIBW 2992, biricodar, brostallicin, bryostatin, buthionine sulfoximine, CBV (chemotherapy), calyculin, non-specific cell cycle antitumor agents, dichloroacetic acid, discodermolide, elsamitrucin, enocytabine, epothilone, eribulin, everolimus, exatecan, exisulind, ferruginol, forodesine, fosfestrol, chemotherapy regimen ICE, IT-101, imexone, imiquimod, indolocarbazole, irofulven, laniquidar, larotaxel, lenalidomide, lucanton, lurtothecan, mafosfamide, mitozolomide, nafoxidine, nedaplatin, olaparib, ortataxel, PAC-1, pawpaw extract, pixantrone, proteasome inhibitor, rebeccamycin, resiquimod, rubitecan, SN-38, salinosporamide A, sapacitabine, Stanford V, swainsonine, talaporfin, tariquidar, tegafururacil, temodar, tesetaxel, triplatinum tetranitrate, tris-(2-chloroethyl)amine, troxacitabine, uramustine, vadimezan, vinflunine, ZD6126 or zosuquidar.

The present invention further relates to a method of using compounds or pharmaceutical compositions provided herein in combination with radiation therapy to inhibit abnormal cell growth or treat a hyperproliferative disorder in a mammal. Techniques for administering radiation therapy are known in the art, and these techniques can be used in the combination therapy described herein. Administration of a compound of the present invention in a given combination therapy can be determined as described herein.

Radiation therapy can be administered through one of several modalities or a combination of modalities, including, but not limited to, external beam radiation therapy, internal radiation therapy, implant-delivered radiation therapy, stereotactic radiosurgery, systemic radiation therapy, radiation therapy, and permanent or temporary interstitial brachytherapy. As used herein, the term brachytherapy refers to radiation therapy delivered by spatially confined radioactive material introduced into the body at or near a tumor or other site of proliferative tissue due to disease. The term is intended to include, without limitation, exposure to radioactive isotopes (e.g., At-211, I-131, I-125, Y-90, Re-186, Re-188, Sm-153, Bi-212, P-32 and radioactive isotopes Lu). Suitable radiation sources for use as a cell enhancer of the present invention include both solids and liquids. By way of non-limiting example, the radiation source may be a radionuclide such as I-125, I-131, Yb-169, Ir-192 as a solid source, I-125 as a solid source, or other radionuclides that emit photons, beta particles, gamma radiation or other rays that have a therapeutic effect. The radioactive material may also be a fluid prepared from any solution of radionuclide(s), for example a solution of I-125 or I-131, or the radioactive fluid may be prepared using a slurry of a suitable fluid containing small particles of solid radionuclides such as like Au-198, Y-90. Moreover, radionuclide(s) may be included in the gel or radioactive microspheres.

- 38 044387

The compounds or pharmaceutical compositions of the present invention can be used in combination with an amount of one or more substances selected from antiangiogenic agents, signal transduction inhibitors, antiproliferative agents, glycolysis inhibitors or autophagy inhibitors.

Antiangiogenic agents such as MMP-2 (matrix metalloproteinase 2) inhibitors, MMP-9 (matrix metalloproteinase 9) inhibitors and COX-11 (cyclooxygenase 11) inhibitors can be used in combination with the compound of the present invention and the pharmaceutical compositions described herein. document. Antiangiogenic agents include, for example, rapamycin, temsirolimus (CCI-779), everolimus (RAD001), sorafenib, sunitinib and bevacizumab. Examples of useful COX-II inhibitors include alecoxib, valdecoxib and rofecoxib. Examples of useful matrix metalloproteinase inhibitors are described in WO 96/33172, WO 96/27583, European Patent Application Publication EP 0818442, European Patent Application Publication EP 1004578, WO 98/07697, WO 98/03516, WO 98/34918, WO 98 /34915, WO 98/33768, WO 98/30566, publication of the European patent application 606046, publication of the European patent application 931788, WO 90/05719, WO 99/52910, WO 99/52889, WO 99/29667, WO 99/ 07675, European Patent Application Publication EP 1786785, European Patent Application Publication No. EP 1181017, US Patent Application Publication No. US20090012085, US Patent Application Publication US5863949, US Patent Application Publication US5861510 and European Patent Application Publication EP 0780386, all of which are incorporated herein by reference in their entirety. Preferred inhibitors of MMP-2 and MMP-9 are those that have little or no MMP-1 inhibitory activity. More preferred are inhibitors that selectively inhibit MMP-2 and/or AMP-9 over other matrix metalloproteinases (i.e. MAP-1, MMP-3, MMP-4, MMP-5, MMP-6, MMP- 7, MMP-8, MMP-10, MMP-11, MMP-12 and MMP-13). Some specific examples of MMP inhibitors useful in the present invention are AG-3340, RO 32-3555 and RS 13-0830.

The compounds of the present invention can also be used in combination therapy with other anticancer agents, such as acemannan, aclarubicin, aldesleukin, alemtuzumab, alitretinoin, altretamine, amifostine, aminolevulinic acid, amrubicin, amsacrine, anagrelide, anastrozole, ANCER, ancestim, arglabin, trioxide arsenic, BAM 002 (Novelos), bexarotene, bicalutamide, broxuridine, capecitabine, celmoleukin, cetrorelix, cladribine, clotrimazole, cytarabine oxphosphate, DA 3030 (Dong-A), daclizumab, denileukin diftitox, deslorelin, dexrazoxane, dilazep, docetaxel, docosanol , doxercalciferol, doxifluridine, doxorubicin, bromocriptine, carmustine, cytarabine, fluorouracil, HIT-diclofenac, interferon alpha, daunorubicin, doxorubicin, tretinoin, edelfosine, edrecolomab, eflornithine, emitefur, epirubicin, epoetin beta, etoposide phosphate, exemestane, exisulind, fadrozole , filgrastim , finasteride, fludarabin phosphate, foreestan, fotemustine, nitrate gallium, hemcitabin, gemtuzumab zogiacin, combination of hymeresil/ocherceil/tegafur, glycopine, gosers, heptaplatin, human chorionic gonadotropin, germinal alpha-phytrotein of human Bitsin, (Imikimod, Interferon alpha, natural interferon alpha, interferon alpha-2, interferon alpha-2a, interferon alpha-2b, interferon alpha-N1, interferon alpha-n3, interferon alphacon-1, interferon alpha, natural interferon beta, interferon beta-1a , interferon beta-1b, interferon-gamma, natural interferon gamma-1a, interferon gamma-1b, interleukin-1 beta, yobenguane, irinotecan, irsogladin, lanreotide, LC 9018 (Yakult), leflunomide, lenograstim, lentinan sulfate, letrozole, leukocyte alpha-interferon, leuprorelin, levamisole+fluorouracil, liarozole, lobaplatin, lonidamine, lovastatin, mazoprocol, melarsoprol, metoclopramide, mifepristone, miltefosine, mirimostim, non-complementary double-stranded RNA, mitoguazone, mitolactol, mitoxantrone, molgramostim, nafarelin, na Loxon + pentazocine, nartograstim, nedaplatin, nilutamide, noscapine, new erythropoiesis-stimulating protein, octreotide NSC 631570, oprelvequin, osaterone, oxaliplatin, paclitaxel, pamidronic acid, pegaspargase, peginterferon-alpha-2b, sodium pentosan polysulfate, pentostatin, picibanil, pyrarubicin, antithymocyte rabbit polyclonal antibodies, polyethylene glycol interferon-alpha-2a, sodium porfimer, raloxifene, raltitrexed, razburicase, rhenium Re 186 etidronate, RII retinamide, rituximab, romurtide, samarium (153 Sm) lexidronam, sargramostim, sisofiran, sobuzoxan, sonermin, strontium-89 chloride, suramin, Tazonordin, Tazaroten, Tegafur, Temoroportin, Temo-Zolomide, Tencere, Tetrylordecakside, Talidomide, Timalfazin, Tirotropin Alpha, Topotoktokan, Toremiafen, Tomomomab-Iiod 131, Trustuzumab, Treosepan, Tretinoin, Trilon, Trimetrexate, Trimettorlin Rosa Tumor Alpha, Natural, Ubenimex , bladder cancer vaccine, Maruyama vaccine, melanoma lysate vaccine, valrubicin, verteporfin, vinorelbine, virulysin, zinostatin stimalamer, or zoledronic acid; abarelix; AE 941 (Aeterna), ambamustine, antisense oligonucleotide, bcl-2 (Genta), APC 8015 (Dendreon), cetuximab, decitabine, dexaminoglutethimide, diaziquon, EL 532 (Elan), EM 800 (Endorecherche), eniluracil, etanidazole, fenretinide, filgrastim SD01 (Amgen), fulvestrant, halocitabine, gastrin 17 immunogen, HLA-B7 for gene therapy (Vical), granulocyte macrophage colony-stimulating factor, histamine dihydrochloride, ibritumomab tiuxetan, ilomastat, IM 862 (Cytran), interleukin-2,

- 39 044387 iproxifen, LDI 200 (Milkhaus), leridistim, lintuzumab, monoclonal antibody CA 125 (Biomira), anticancer monoclonal antibodies (Japan Pharmaceutical Development), monoclonal antibodies HER-2 and Fc (Medarex), idiotypic monoclonal antibody 105AD7 (CRC Technology ), idiotypic monoclonal antibody CEA (Trilex), monoclonal antibody LYM-1-iodine 131 (Techniclone), monoclonal antibody to polymorphic epithelial mucin, yttrium-labeled 90 (Antisoma), marimastat, menogaryl, mitumomab, motexafin gadolinium, MX 6 (Galderma) , nelarabine, nolatrexed, protein P 30, pegvisomant, pemetrexed, porfiromycin, prinomastat, RL 0903 (Shire), rubitecan, satraplatin, sodium phenylacetate, sparphosic acid, SRL 172 (SR Pharma), SU 5416 (SUGEN), TA 077 (Tanabe ), tetrathiomolybdate, thaliblastin, thrombopoietin, ethylethiopurpurin stannous, tirapazamine, cancer vaccine (Biomira), melanoma vaccine (NYU), melanoma vaccine (Sloan-Kettering Institute), melanoma oncolysate vaccine (New York Medical College), vaccine based on viral melanoma cell lysates (Royal Newcastle Hospital) or valspodar.

The compounds of the present invention can additionally be used with VEGFR inhibitors. Other compounds described in the following patents and patents can be used in combined therapy: US 6258812, US 2003/0105091, Wo 01/37820, US 6235764, Wo 01/32651, US 6630500, US 6515004, US 6713485, US 552111 84 , US 5770599, US 5747498, WO 02/68406, WO 02/66470, WO 02/55501, WO 04/05279, WO 04/07481, WO 04/07458, WO 04/09784, WO 02/59110, WO 9 9/ 45009, WO 00/59509, WO 99/61422, US 5990141, WO 00/12089 and WO 00/02871.

In some embodiments, the combination provides a composition of the present invention in combination with at least one antiangiogenic agent. Such agents include, without limitation, in vitro synthetically produced chemical compositions, antibodies, antigen-binding sites, radionuclides, as well as combinations and conjugates thereof. The agent may be an agonist, antagonist, allosteric modulator, toxin or, more generally, may act in an inhibitory or stimulatory manner on its target (eg, activation or inhibition of a receptor or enzyme) and thereby promote cell death or arrest cell growth.

Exemplary antiangiogenic agents include ERBITUX™ (IMC-C225), KDR (kinase domain receptor) inhibitory agents (e.g., antibodies and antigen binding sites that specifically bind to the receptor kinase domain), anti-VEGF agents (e.g., antibodies and antigen-binding sites that specifically bind to VEGF, or soluble VEGF receptors or a ligand-binding site thereof), such as AVASTIN™ or VEGF-TRAP™, and VEGF receptor antibody agents (e.g., antibodies or antigen-binding sites that specifically bind to agents that inhibit EGFR (e.g., antibodies or antigen-binding sites that specifically bind to it), such as Vectibix (panitumumab), IRESSA™ (gefitinib), TARCEVA™ (erlotinib), anti-Ang1 and Ang2 anti-EGFR agents (for example, antibodies or antigen-binding sites that specifically bind to it or their receptors, for example Tie2/Tek), as well as inhibitory agents that are antibodies to Tie2 kinase (for example, antibodies or antigen-binding sites that specifically bind to it) . The pharmaceutical compositions of the present invention may also contain one or more agents (e.g., antibodies, antigen-binding sites, or soluble receptors) that specifically bind to and inhibit the activity of growth factors, such as hepatocyte growth factor (HGF, also known as scattering factor) antagonists and antibodies or antigen-binding sites that specifically bind to its c-met receptor.

Other anti-angiogenic agents include Campath, IL-8, B-FGF, Tek antagonists (Ceretti et al., US Patent Application Publication No. 2003/0162712, US Patent No. 6,413,932), anti-TWEAK antibody agents (eg, specifically binding antibodies or antigen binding sites or soluble receptor antagonists TWEAK, see Wiley, US Pat. No. 6,727,225), ADAM disintegrin domain as an antagonist of integrin binding to its ligands (Fanslow et al., US Patent Application Publication No. 2002/0042368), specifically binding anti-eph receptor antibodies and/or anti-ephrin antibodies or antigen binding sites (US Pat. Nos. 5,981,245, 5,728,813, 5,969,110, 6,596,852, 6,232,447, 6,057,124 and members thereof) and antagonists that are antibodies to PDGF-BB (e.g., specifically binding antibodies or antigen-binding sites), as well as antibodies or antigen-binding sites that specifically bind to PDGF-BB ligands, and agents that are inhibitors of PDGFR kinase (for example, antibodies or antigen-binding sites that specifically bind to it).

Additional antiangiogenic/antitumor agents include SD-7784 (Pfizer, USA); cilengitide (Merck KGaA, Germany, EPO 770622); pegaptanib octasodium (Gilead Sciences, USA); alfastatin (BioActa, UK); M-PGA (Celgene, USA, US 5712291); ilomastat (Arriva, USA, US 5892112); emaxanib (Pfizer, USA, US 5792783); vatalanib (Novartis, Switzerland);

- 40 044387

2-methoxyestradiol (EntreMed, USA); TLC ELL-12, (Elan, Ireland); anekortav acetate (Alcon, USA); monoclonal antibody alpha-0148 (Amgen, USA); CEP-7055 (Cephalon, USA); monoclonal antibody to Vn (Crucell, Netherlands) DAC:antiangiogen (ConjuChem, Canada); angiocidin (InKine Pharmaceutical, USA); KM-2550 (Kyowa Hakko, Japan); SU-0879 (Pfizer, USA); CGP-79787 (Novartis, Switzerland, EP 970070); ARGENT technology (Ariad, USA); YIGSR-Stealth, (Johnson & Johnson, USA); fibrinogen fragment-E (BioActa, UK); angiogenesis inhibitor (Trigen, UK); TVS-1635 (Encysive Pharmaceuticals, USA); SC-236 (Pfizer, USA); AVT-567 (Abbott, USA); metastatin (EntreMed, USA); angiogenesis inhibitor (Tripep, Sweden); maspin (Sosei, Japan); 2-methoxyestradiol (Oncology Sciences Corporation, USA); ER-68203-00 (IVAX, USA); benefin (Lane Labs, USA); Tz-93 (Tsumura, Japan); TAN-1120 (Takeda, Japan); FR-111142 (Fujisawa, Japan, JP 02233610); platelet factor 4 (RepliGen, USA, EP 407122); vascular endothelial growth factor antagonist (Borean, Denmark); bevacizumab (pINN), (Genentech, USA); angiogenesis inhibitors (SUGEN, USA); XL 784 (Exelixis, USA); XL 647 (Exelixis, USA); monoclonal antibody, second generation alpha5beta3 integrin (Applied Molecular Evolution, USA, and Medlmmune, USA); a product for gene therapy of retinopathy (Oxford BioMedica, UK); enzastaurine hydrochloride (USAN), (Lilly, USA); CEP 7055, (Cephalon, USA, and Sanofi-Synthelabo, France); BC 1 (Genoa Institute of Cancer Research, Italy); angiogenesis inhibitor (Alchemia, Australia); VEGF antagonist (Regeneron, USA); antiangiogenic drugs based on rBPI 21 and BPI (HOMA, USA); PI 88 (Progen, Australia); cilengitide (pINN), (Merck KGaA, Germany; Technical University of Munich, Germany, Scripps Clinic and Research Foundation, USA); cetuximab (INN), (Aventis, France); AVE 8062 (Ajinomoto, Japan); AS 1404 (Cancer Research Laboratory, New Zealand); SG 292 (Telios, USA); endostatin (Boston Children's Hospital, USA); ATN 161 (Attenuon, USA); angiostatin (Boston Children's Hospital, USA); 2-methoxyestradiol (Boston Children's Hospital, USA); ZD 6474 (AstraZeneca, UK); ZD 6126 (Angiogene Pharmaceuticals, UK); PPI 2458 (Praecis, USA); AZD 9935 (AstraZeneca, UK); AZD 2171 (AstraZeneca, UK); vatalanib (pINN), (Novartis, Switzerland and Schering AG, Germany); tissue factor pathway inhibitors (EntreMed, USA); pegaptanib (Pinn), (Gilead Sciences, USA); xanthorizol, (Yonsei University, South Korea); genetically engineered vaccine, VEGF-2, (Scripps Clinic and Research Foundation, USA); SPV5.2 (Supratek, Canada); SDX 103 (University of California at San Diego, USA); РХ 478 (ProlX, USA); metastatin, (EntreMed, USA); Troponin I, (Harvard University, USA); SU 6668 (SUGEN, USA); OXI 4503 (OXiGENE, USA); o-guanidines (Dimensional Pharmaceuticals, USA); motuporamine C (University of British Columbia, Canada); CDP 791 (Celltech Group, UK); atiprimod (pINN), (GlaxoSmithKline, UK); E 7820 (Eisai, Japan); CYC 381 (Harvard University, USA); AE 941 (Aeterna, Canada); angiogenesis vaccine (EntreMed, USA); urokinase plasminogen activator inhibitor (Dendreon, USA); oglufanide (pINN), (Melmotte, USA); HIF-1 alpha inhibitors (Xenova, UK); CEP 5214 (Cephalon, USA); BAY RES 2622 (Bayer, Germany); angiocidin (InKine, USA); A6 (Angstrom, USA); KR 31372 (Korea Chemical Technology Research Institute, South Korea); GW 2286 (GlaxoSmithKline, UK); UNT 0101 (ExonHit, France); CP 868596 (Pfizer, USA); CP 564959 (OSI, USA); CP 547632 (Pfizer, USA); 786034 (GlaxoSmithKline, UK); KRN 633 (Kirin Brewery, Japan); intraocular drug delivery system, 2-methoxyestradiol (EntreMed, USA); Anginex, (Maastricht University, the Netherlands, and the University of Minnesota, USA); ABT 510 (Abbott, USA); AAL 993 (Novartis, Switzerland); VEGI (ProteomTech, USA); tumor necrosis factor alpha inhibitors (National Institute on Aging, USA); SU 11248 (Pfizer, USA and SUGEN, USA); ABT 518 (Abbott, USA); YH16 (Yantai Rongchang, China); S-3APG (Boston Children's Hospital, USA, and EntreMed, USA); monoclonal antibody, KDR (ImClone Systems, USA); monoclonal antibody, alpha5beta1 (Protein Design, USA); KDR kinase inhibitor (Celltech Group, UK, and Johnson & Johnson, USA); GFB 116 (University of South Florida, USA, and Yale University, USA); CS 706 (Sankyo, Japan); prodrug combretastatin A4 (Arizona State University, USA); chondroitinase AC (IBEX, Canada); BAY RES 2690 (Bayer, Germany); AGM 1470 (Harvard University, USA; Takeda, Japan, and TAR, USA); AG 13925 (Agouron, USA); tetrathiomolybdate (University of Michigan, USA); GCS 100 (Wayne University, USA), CV 247 (Ivy Medical, UK); CKD 732 (Chong Kun Dang, South Korea); monoclonal antibody, vascular endothelial growth factor (Xenova, UK); Irsogladine (INN), (Nippon Shinyaku, Japan); RG 13577 (Aventis, France); WX 360 (Wilex, Germany); squalamine (pINN), (Genaera, USA); RPI 4610 (Sirna, USA); cancer therapy (Marinova, Australia); heparanase inhibitors (InSight, Israel); KL 3106 (Kolon, South Korea); honokiol (Emory University, USA); ZK CDK (Schering AG, Germany); ZK Angio (Schering AG, Germany); ZK 229561 (Novartis, Switzerland, and Schering AG, Germany); XMP 300 (HOMA, USA); VGA 1102 (Taisho, Japan); VEGF receptor modulators (Pharmacopeia, USA); VE-cadherin-2 antagonists (ImClone Systems, USA); vasostatin (National Institute of Health, USA); Flk-1 vaccine (ImClone Systems, USA); TZ 93 (Tsumura, Japan); tumstatin (Beth Israel Hospital, USA); shortened soluble FLT 1 (vascular endothelial growth factor receptor 1), (Merck & Co, USA); Tie-2 ligands (Regeneron, USA) and thrombospondin 1 inhibitor (Allegheny

- 41 044387

Health, Education and Research Foundation, USA).

Autophagy inhibitors include, but are not limited to, chloroquine, 3-methyladenine, hydroxychloroquine (Plaquenil™), bafilomycin A1, 5-amino-4-imidazole carboxamide riboside (AICAR), algal toxins that inhibit autophagy that inhibit protein phosphatases type 2A or type 1, cAMP analogues and drugs that increase cAMP levels, such as adenosine, LY204002, N ⁶ -mercaptopurine riboside and vinblastine. In addition, antisense or siRNA that inhibits the expression of proteins, including but not limited to ATG5 (which are involved in autophagy), can also be used.

Additional pharmaceutically active compounds/agents that can be used in the treatment of cancers and that can be used in combination with one or more compounds of the present invention include: epoetin alfa, darbepoetin alfa, panitumumab, pegfilgrastim, palifermin, filgrastim, denosumab, ancestim, AMG 102 , AMG 386, AMG 479, AMG 655, AMG 745, AMG 951 and AMG 706 or pharmaceutically acceptable salts thereof.

In some embodiments, a composition provided herein is administered in conjunction with a chemotherapeutic agent. Suitable chemotherapeutic agents may include natural products such as vinca alkaloids (eg, vinblastine, vincristine and vinorelbine), paclitaxel, epidipodophyllotoxins (eg, etoposide and teniposide), antibiotics (eg, dactinomycin (actinomycin D), daunorubicin, doxorubicin and idarubicin), anthracyclines, mitoxantrone, bleomycins, plicamycin (mithramycin), mitomycin, enzymes (for example, L-asparaginase, which systemically metabolizes L-asparagine and deprives it of cells that do not have the ability to synthesize their own asparagine), antiplatelet agents, antiproliferative/antimitotic alkylating agents, such as nitrogen mustards (eg, mechlorethamine, cyclophosphamide and analogues, melphalan and chlorambucil), ethylenimines and methylmelamines (eg, hexamethylmelamine and thiotepa), CDK inhibitors (eg, seliciclib, UCN-01, P1446A-05, PD-0332991, dinaciclib, P27-00, AT-7519, RGB286638 and sCh727965), alkyl sulfonates (e.g. busulfan), nitrosourea derivatives (e.g. carmustine (BCNU) and analogs, as well as streptozocin), trazenes-dacarbazinine (DTIC), antiproliferative/antimitotic antimetabolites such as folic acid analogues (eg, methotrexate), pyrimidine analogues (eg, fluorouracil, floxuridine and cytarabine), purine analogues and related inhibitors (eg, mercaptopurine, thioguanine, pentostatin and 2-chlorodeoxyadenosine), aromatase inhibitors (eg, anastrozole, exemestane and Letrozole) and coordination complexes of platinum (for example, cisplatin and carboplatin), pricarbazine, hydroxymochevin, mitotan, aminoglutetimide, histonezacetylsis inhibitors (HDAC) (for example, trichostatin, sodium butyrate, apicidane, Suberoilanilidhydromic acid, Vorinostat, LBH 58 9, Romidepsin, Acy-1215 and panobinostat), mTor inhibitors (eg, temsirolimus, everolimus, ridaforolimus, and sirolimus), KSP(Eg5) inhibitors (eg, Array 520), DNA binding agents (eg, Zalypsis), PI3K delta inhibitor (eg, GS-1101 and TGR-1202), PI3K delta and gamma inhibitor (eg, CAL-130), multikinase inhibitor (eg, TG02 and sorafenib), hormones (eg, estrogen), and hormone agonists such as luteinizing hormone releasing factor (LHRH) agonists ( e.g., goserelin, leuprolide, and triptorelin), BAFF neutralizing antibody (e.g., LY2127399), IKK inhibitors, p38MAPK inhibitors, anti-IL-6 antibodies (e.g., CNTO328), telomerase inhibitors (e.g., GRN 163L), aurorakinase inhibitors (e.g., MLN8237), cell surface monoclonal antibodies (eg, anti-CD38 antibodies (HUMAX-CD38), anti-CS1 antibodies (eg, elotuzumab), HSP90 inhibitors (eg, 17 AAG and KOS 953), P13K/Akt inhibitors (eg, perifosine), Akt inhibitor (eg, GSK-2141795), PKC inhibitors (eg, enzastaurine), FTI species (eg, Zarnestra™), anti-CD138 antibodies (eg, BT062), Torc1/2 specific kinase inhibitor (eg, INK128), kinase inhibitor (eg, GS-1101), ER/UPR targeting agent (eg, MKS-3946), cFMS inhibitor (eg, ARRY-382), JAK1/2 inhibitor (eg, CYT387), PARP inhibitor (eg, olaparib and veliparib ( ABT-888)), BCL-2 antagonist. Other chemotherapeutic agents may include mechlorethamine, camptothecin, ifosfamide, tamoxifen, raloxifene, gemcitabine, navelbine, sorafenib, or any analog or derivative of the above.

The compounds of the present invention can also be used in combination with radiation therapy, hormonal therapy, surgery and immunotherapy, such therapies being well known to those skilled in the art.

In some embodiments, a pharmaceutical composition provided herein is co-administered with a steroid. Suitable steroids may include, but are not limited to, 21-acetoxypregnenolone, alklometasone, algestone, amcinonide, beclomethasone, betamethasone, budesonide, chlorprednisone, clobetasol, clocortolone, cloprednol, corticosterone, cortisone, cortivasol, deflazacort, desonide, deoxymethasone, dexamethasone, diflo razone, diflucortolone, difuprednate, enoxolone, fluazacort, flucloronide, flumethasone, flunisolide, fluopinolone acetonide, fluocinonide, fluocortin butyl, fluocortolone, fluorometholone, fluperolone acetate, fluprednidene acetate, fluprednisolone,

- 42 044387 fluranrenolide, fluticasone propionate, formocortal, galcinonide, halobetasol, halomethasone, hydrocortisone, loteprednol etabonate, mazipredone, medrisone, meprednisone, methylprednisolone, mometasone furoate, paramethasone, prednicarbate, prednisolone, prednisolone 25-diethyl aminoacetate, prednisolone sodium phosphate, prednisone, prednival , prednilidene, rimexolone, thixocortol, triamcinolone, triamcinolone acetonide, triamcinolone benetonide, triamcinolone hexacetonide, as well as their salts and/or derivatives. In a specific embodiment, the compounds of the present invention can also be used in combination with additional pharmaceutically active agents that treat nausea. Examples of agents that can be used to treat nausea include dronabinol, granisetron, metoclopramide, ondansetron and prochlorpemazine or pharmaceutically acceptable salts thereof.

The compounds or pharmaceutical compositions of the present invention can also be used in combination with an amount of one or more substances selected from EGFR inhibitors, MEK inhibitors, PI3K inhibitors, AKT inhibitors, TOR inhibitors and immune therapeutics, including anti-PD-1 antibody agents , anti-PDL-1 antibodies, anti-CTLA4 antibodies, anti-LAG1 antibodies and anti-OX40 antibodies, GITR agonists, CAR-T cells and BiTE varieties.

EGFR inhibitors include, but are not limited to, small molecule antagonists, antibody-based inhibitors, or specific antisense nucleotides or siRNAs. Useful antibody-based EGFR inhibitors include cetuximab (Erbitux), panitumumab (Vectibix), zalutumumab, nimotuzumab, and matuzumab. Small molecule EGFR antagonists include gefitinib, erlotinib (Tarceva) and, more recently, lapatinib (Tykerb). See, for example, Yan L., et. al., Pharmacogenetics and Pharmacogenomics In Oncology Therapeutic Antibody Development, BioTechniques, 2005; 39(4):565-8 and Paez J.G., et. al., EGFR Mutations In Lung Cancer Correlation With Clinical Response To Gefitinib Therapy, Science, 2004; 304(5676):1497-500.

Non-limiting examples of small molecule EGFR inhibitors include any of the EGFR inhibitors described in the following patent publications, and all pharmaceutically acceptable salts and solvates of these EGFR inhibitors: European patent application EP 520722, published December 30, 1992; European patent application EP 566226, published October 20, 1993; publication of PCT international application WO 96/33980, published on October 31, 1996; US Patent No. 5,747,498, issued May 5, 1998; publication of PCT international application WO 96/30347, published on October 3, 1996; European patent application EP 787772, published 6 August 1997; publication of PCT international application WO 97/30034, published on August 21, 1997; publication of PCT international application WO 97/30044, published on August 21, 1997; publication of PCT international application WO 97/38994, published on October 23, 1997; publication of PCT international application WO 97/49688, published on December 31, 1997; European patent application EP 837063, published April 22, 1998; publication of PCT international application WO 98/02434, published on January 22, 1998; publication of PCT international application WO 97/38983, published on October 23, 1997; publication of PCT international application WO 95/19774, published on July 27, 1995; publication of PCT international application WO 95/19970, published on July 27, 1995; publication of PCT international application WO 97/13771, published April 17, 1997; publication of PCT international application WO 98/02437, published on January 22, 1998; publication of PCT international application WO 98/02438, published on January 22, 1998; publication of PCT international application WO 97/32881, published on September 12, 1997; German patent application DE 19629652, published January 29, 1998; publication of PCT international application WO 98/33798, published on August 6, 1998; publication of PCT international application WO 97/32880, published on September 12, 1997; publication of PCT international application WO 97/32880, published on September 12, 1997; European patent application EP 682027, published November 15, 1995; publication of PCT international application WO 97/02266, published on January 23, 197; publication of PCT international application WO 97/27199, published on July 31, 1997; publication of PCT international application WO 98/07726, published on February 26, 1998; publication of PCT international application WO 97/34895, published September 25, 1997; publication of PCT international application WO 96/31510', published on October 10, 1996; publication of PCT international application WO 98/14449, published April 9, 1998; publication of PCT international application WO 98/14450, published on April 9, 1998; publication of PCT international application WO 98/14451, published on April 9, 1998; publication of PCT international application WO 95/09847, published April 13, 1995; publication of PCT international application WO 97/19065, published on May 29, 1997; publication of PCT international application WO 98/17662, published April 30, 1998; US Patent No. 5,789,427, issued August 4, 1998; US Patent No. 5650415, issued July 22, 1997; US Patent No. 5656643, issued August 12, 1997; publication of international application filed under PCT WO 99/35146, pub.

- 43 044387 jubilant July 15, 1999; publication of PCT international application WO 99/35132, published on July 15, 1999; publication of PCT international application WO 99/07701, published on February 18, 1999; and publication of PCT-filed international application WO 92/20642, published November 26, 1992. Additional non-limiting examples of small molecule EGFR inhibitors include any of the EGFR inhibitors described in Traxler, P., 1998, Exp. Opin. Ther. Patents 8(12):1599–1625.

Antibody-based EGFR inhibitors include any anti-EGFR antibody or antibody fragment that can partially or completely block activation of EGFR by its natural ligand. Non-limiting examples of antibody-based EGFR inhibitors include those described in Modjtahedi, H., et al., 1993, Br. J Cancer 67:247-253; Teramoto, T., et al., 1996, Cancer, 77:639-645; Goldstein et al., 1995, Clin. Cancer Res. 1:1311-1318; Huang, S.M., et al., 1999, Cancer Res. 15:59(8):1935-40 and Yang, X., et al., 1999, Cancer Res. 59:1236–1243. Thus, the EGFR inhibitor may be a monoclonal antibody Mab E7.6.3 (Yang, 1999 supra) or Mab C225 (ATCC Accession No. HB-8508) or an antibody or antibody fragment having binding specificity thereto.

MEK inhibitors include, but are not limited to, CI-1040, AZD6244, PD318088, PD98059, PD334581, RDEA119, ARRY-142886, ARRY-438162, and PD-325901.

PI3K inhibitors include, but are not limited to:

wortmannin, 17-hydroxywortmannin analogues described in WO 06/044453,

4-[2-(1H-Indazol-4-yl)-6-[[4-(methylsulfonyl)piperazin-1-yl]methyl]thieno[3,2-d]pyrimidin-4yl]morpholine (also known as GDC 0941 and described in PCT publications No. WO 09/036082 and WO 09/055730),

2-methyl-2-[4-[3-methyl-2-oxo-8-(quinolin-3-yl)-2,3-dihydroimidazo[4,5-c]quinolin-1yl]phenyl]propionitrile (also known as BEZ 235 or NVP-BEZ 235 and described in PCT publication No. WO 06/122806), (S)-1-(4-((2-(2-aminopyrimidin-5-yl)-7-methyl-4- morpholinothieno[3,2-d]pyrimidin-6yl)methyl)piperazin-1-yl)-2-hydroxypropan-1-one (described in PCT publication No. WO 2008/070740),

LY294002 (2-(4-morpholinyl)-8-phenyl-4H-1-benzopyran-4-one, available from Axon Medchem),

PI 103 hydrochloride (3-[4-(4-morpholinylpyrido[3',2':4,5]furo[3,2-d]pyrimidin-2-yl]phenol hydrochloride, available from Axon Medchem),

PIK 7 5 (N'-[(1 E)-(6-bromimidazo[1,2-a]pyridin-3-yl)methylene]-N,2-dimethyl-5-nitrobenzenesulfonohydrazide hydrochloride, available from Axon Medchem),

PIK 90 (N-(7,8-dimethoxy-2,3-dihydroimidazo[1,2-c]quinazolin-5-yl)nicotinamide, available from Axon Medchem),

GDC-0941 bis-mesylate (2-(1H-indazol-4-yl)-6-(4-methanesulfonylpiperazin-1-ylmethyl)-4morpholin-4-yl-thieno[3,2-d]pyrimidine bismesylate, available from Axon Medchem),

AS-252424 (5-[1-[5-(4-fluoro-2-hydroxyphenyl)furan-2-yl]-meth-(Z)-ylidene]thiazolidine-2,4-dione, available from Axon Medchem), And

TGX-221 (7-methyl-2-(4-morpholinyl)-9-[1-(phenylamino)ethyl]-4H-pyrido[1,2-a]pyrimidin-4-one, available from Axon Medchem),

XL-765 and XL-147.

Other PI3K inhibitors include demethoxyviridine, perifosine, CAL101, PX-866, BEZ235, SF1126, INK1117, IPI-145, BKM120, XL147, XL765, palomide 529, GSK1059615, ZSTK474, PWT33597, IC87114, TG100-115, CAL263, PI-103 , GNE-477, CUDC-907 and AEZS-136.

AKT inhibitors include, but are not limited to, Akt-1-1 (inhibits Akt1) (Barnett et al. (2005), Biochem. J., 385 (Pt. 2), 399-408); Akt-1-1,2 (inhibits Ak1 and 2) (Barnett et al. (2005), Biochem. J. 385 (Pt. 2), 399-408); API-59CJ-Ome (eg, Jin et al. (2004), Br. J. Cancer, 91, 1808-12); 1H-imidazo[4,5c]pyridinyl compounds (eg WO 05011700); indole-3-carbinol and derivatives thereof (eg, US Pat. No. 6,656,963; Sarkar and Li (2004), J. Nutr. 134(12 Suppl), 3493S-3498S); perifosine (eg, interferes with membrane localization of Akt; Dasmahapatra et al. (2004) Clin. Cancer Res. 10(15), 524252, 2004); phosphatidylinositol ester, lipid analogues (e.g. Gills and Dennis (2004), Expert. Opin. Investig. Drugs 13, 787-97) and triciribine (TCN or API-2 or NCI ID: NSC 154020; Yang et al. (2004) , Cancer Res. 64, 4394-9).

TOR inhibitors include, but are not limited to, inhibitors that contain AP-23573, CCI-779, everolimus, RAD-001, rapamycin, temsirolimus, ATP-competitive TORC1/TORC2 inhibitors including PI-103, PP242, PP30 and Torin 1. Others TOR inhibitors in the FKBP12 enhancer; rapamycins and their derivatives, including: CCI-779 (temsirolimus), RAD001 (everolimus; WO 9409010) and AP23573; rapalogues, for example, as disclosed in WO 98/02441 and WO 01/14387, for example AP23573, AP23464 or AP23841; 40-(2-hydroxyethyl)rapamycin, 40-[3-hydroxy(hydroxymethyl)methylpropanoate]rapamycin (also called CC1779), 40-epi-(tetrazolyte)rapamycin (also called ABT578), 32-deoxorapamycin, 16-pentynyloxy-32 (S)-dihydrorapamycin and other derivatives disclosed in WO 05/005434; derivatives disclosed in US Patent No. 5258389, WO 94/090101, Wo 92/05179, US Patent Nos. 5118677, 5118678, 5100883, 5151413, 5120842, WO 93/111130, WO 94/02136,

- 44 044387

WO 94/02485, WO 95/14023, WO 94/02136, WO 95/16691, WO 96/41807, WO 96/41807 and US patent No. 5256790; phosphorus-containing rapamycin derivatives (eg WO 05016252);

4H-1-benzopyran-4-one derivatives (eg, US Provisional Patent Application No. 60/528,340).

Immune therapeutic agents include, but are not limited to, anti-PD-1 antibody agents, anti-PDL-1 antibody agents, anti-CTLA-4 antibody agents, anti-LAG1 antibody agents, and anti-LAG1 antibody agents. are antibodies to OX40. Exemplary anti-PD-1 antibodies and methods of use thereof are described in Goldberg et al., Blood, 110(1):186-192 (2007), Thompson et al., Clin. Cancer Res. 13(6):1757-1761 (2007) and Korman et al., International Application No. PCT/JP2006/309606 (Publication No. WO 2006/121168 A1), each of which is expressly incorporated by reference herein. Includes: Yervoy™ (ipilimumab) or tremelimumab (CTLA-4), galiximab (B7.1), BMS-936558 (PD-1), MK-3475 (PD-1), AMP224 (B7DC), BMS-936559 (to B7-H1), MPDL3280A (to B7-H1), MEDI-570 (to ICOS), AMG557 (to B7N2), MGA271 (to B7N3), IMP321 (to LAG-3), BMS-663513 ( anti-CD137), PF-05082566 (anti-CD137), CDX-1127 (anti-CD27), anti-OX40 antibodies (Providence Health Services), huMAbOX40L (anti-OX40L), atacicept (anti-TACI), CP-870893 (anti-CD40), lucatumumab (anti-CD40), dacetuzumab (anti-CD40), muromonab-CD3 (anti-CD3), ipilumumab (anti-CTLA-4). Immune therapeutics also include genetically engineered T cells (eg, CAR-T cells) and bispecific antibodies (eg, BiTE).

GITR agonists include, but are not limited to, GITR fusion proteins and anti-GITR antibodies (eg, bivalent anti-GITR antibodies), such as the GITR fusion protein described in US Pat. No. 611,1090Box.c, EP No. 090505B1, US Pat. No. 8,586,023, PCT - Publication Nos. WO 2010/003118 and 2011/090754, or the anti-GITR antibody described, for example, in US Patent No. 7025962, European Patent No. 1947183B1, US Patent No. 7812135, US Patent No. 8388967, US Patent No. 8591886, European com patent No. EP 1866339, PCT Publication No. WO 2011/028683, PCT Publication No. WO 2013/039954, PCT Publication No. WO 2005/007190, PCT Publication No. WO 2007/133822, PCT Publication No. WO 2005/055808, PCT - Publication No. WO 99/40196, PCT Publication No. WO 2001/03720, PCT Publication No. WO 99/20758, PCT Publication No. WO 2006/083289, PCT Publication No. WO 2005/115451, US Patent No. 7618632 and PCT -publication no. WO 2011/051726.

In some embodiments, a compound having the structure

or a stereoisomer thereof, an atropisomer thereof, a pharmaceutically acceptable salt thereof, a pharmaceutically acceptable salt thereof of a stereoisomer, or a pharmaceutically acceptable salt thereof of an atropisomer, is administered to a patient in need thereof, a therapeutically effective amount of an additional pharmaceutically active compound, for example cytarabine, a proteasome inhibitor such as carfilzomib or oprosomib, a BCl-2 inhibitor such as venetoclax, an MCl-1 inhibitor such as AMG-176, a monoclonal antibody such as daratumumab, and an imide-based immunomodulatory drug (IMiD) such as thalidomide, lenalidomide, pomalidomide and apremilast.

In some embodiments, a compound having the structure

or a stereoisomer thereof, an atropisomer thereof, a pharmaceutically acceptable salt thereof, a pharmaceutically acceptable salt thereof of a stereoisomer, or a pharmaceutically acceptable salt thereof of an atropisomer, is administered to a patient in need thereof, a therapeutically effective amount of an additional pharmaceutically active compound, for example cytarabine, a proteasome inhibitor such as carfilzomib or oprosomib, a BCl-2 inhibitor such as venetoclax, an MCl-1 inhibitor such as AMG-176, a monoclonal antibody such as daratumumab, and an imide-based immunomodulatory drug (IMiD) such as thalidomide, lenalidomide, pomalidomide and apremilast.

In some embodiments, a compound having the structure

- 45 044387 or a stereoisomer thereof, an atropisomer thereof, a pharmaceutically acceptable salt thereof, a pharmaceutically acceptable salt thereof of a stereoisomer or a pharmaceutically acceptable salt thereof of an atropisomer is administered to a patient in need thereof, a therapeutically effective amount of an additional pharmaceutically active compound, for example cytarabine, a proteasome inhibitor such as carfilzomib or oprosomib, a BCl-2 inhibitor such as venetoclax, an MCl-1 inhibitor such as AMG-176, a monoclonal antibody such as daratumumab, and an imide-based immunomodulatory drug (IMiD) such as thalidomide, lenalidomide, pomalidomide, and apremilast .

In some embodiments, a compound having the structure

or a stereoisomer thereof, an atropisomer thereof, a pharmaceutically acceptable salt thereof, a pharmaceutically acceptable salt thereof of a stereoisomer, or a pharmaceutically acceptable salt thereof of an atropisomer, is administered to a patient in need thereof, a therapeutically effective amount of an additional pharmaceutically active compound, for example cytarabine, a proteasome inhibitor such as carfilzomib or oprosomib, a BCl-2 inhibitor such as venetoclax, an MCl-1 inhibitor such as AMG-176, a monoclonal antibody such as daratumumab, and an imide-based immunomodulatory drug (IMiD) such as thalidomide, lenalidomide, pomalidomide and apremilast.

In some embodiments, a compound having the structure

or a stereoisomer thereof, an atropisomer thereof, a pharmaceutically acceptable salt thereof, a pharmaceutically acceptable salt thereof of a stereoisomer, or a pharmaceutically acceptable salt thereof of an atropisomer, is administered to a patient in need thereof, a therapeutically effective amount of an additional pharmaceutically active compound, for example cytarabine, a proteasome inhibitor such as carfilzomib or oprosomib, a BCl-2 inhibitor such as venetoclax, an MCl-1 inhibitor such as AMG-176, a monoclonal antibody such as daratumumab, and an imide-based immunomodulatory drug (IMiD) such as thalidomide, lenalidomide, pomalidomide and apremilast.

In some embodiments, a compound having the structure

FROM

Ll F

K ' N >—' -N OH or a stereoisomer thereof, an atropisomer thereof, a pharmaceutically acceptable salt thereof, a pharmaceutically acceptable salt of a stereoisomer thereof, or a pharmaceutically acceptable salt of an atropisomer thereof is administered to a patient in need thereof, a therapeutically effective amount of an additional pharmaceutically active compound, for example cytarabine, a proteasome inhibitor such as carfilzomib or oprosomib, a BCl-2 inhibitor such as venetoclax, an MCl-1 inhibitor such as AMG-176, a monoclonal antibody such as daratumumab, and an imide-based immunomodulatory drug (IMiD) such as thalidomide , lenalidomide, pomalidomide and apremilast.

In some embodiments, a compound having the structure

or a stereoisomer thereof, an atropisomer thereof, a pharmaceutically acceptable salt thereof, a pharmaceutically acceptable salt thereof of a stereoisomer, or a pharmaceutically acceptable salt thereof of an atropisomer, is administered to a patient in need thereof, a therapeutically effective amount of an additional pharmaceutically active compound, for example cytarabine, a proteasome inhibitor such as carfilzomib or oprosomib, a BCl-2 inhibitor such as venetoclax, an MCl-1 inhibitor such as AMG-176, a monoclonal antibody such as daratumumab, and an imide-based immunomodulatory drug (IMiD) such as thalidomide, lenalidomide, pomalidomide and apremilast.

In some embodiments, a compound having the structure p—

I 'i С| F ⁼ -2) and '

M ^- or a stereoisomer thereof, an atropisomer thereof, a pharmaceutically acceptable salt thereof, a pharmaceutically acceptable salt thereof of a stereoisomer, or a pharmaceutically acceptable salt thereof of an atropisomer is administered to a patient in need thereof, a therapeutically effective amount of an additional pharmaceutically active compound, for example cytarabine, a proteasome inhibitor such as carfilzomib or oprosomib, a BCl-2 inhibitor such as venetoclax, an MCl-1 inhibitor such as AMG-176, a monoclonal antibody such as daratumumab, and an imide-based immunomodulatory drug (IMiD) such as thalidomide, lenalidomide, pomalidomide and apremilast.

In some embodiments, a compound having the structure

W. ' LI F /

N. Ν '— ^;

Ν .— o .—.· ' Ν — or its stereoisomer, its atropisomer, a pharmaceutically acceptable salt thereof, a pharmaceutically acceptable salt of its stereoisomer, or a pharmaceutically acceptable salt of its atropisomer is administered to a patient in need thereof, a therapeutically effective amount of an additional pharmaceutically active compound, e.g. cytarabine, a proteasome inhibitor such as carfilzomib or oprosomib, a BCl-2 inhibitor such as venetoclax, an MCl-1 inhibitor such as AMG-176, a monoclonal antibody such as daratumumab, and an imide-based immunomodulatory drug (IMiD) such such as thalidomide, lenalidomide, pomalidomide and apremilast.

In some embodiments, a compound having the structure

or a stereoisomer thereof, an atropisomer thereof, a pharmaceutically acceptable salt thereof, a pharmaceutically acceptable salt thereof of a stereoisomer, or a pharmaceutically acceptable salt thereof of an atropisomer, is administered to a patient in need thereof, a therapeutically effective amount of an additional pharmaceutically active compound, for example cytarabine, a proteasome inhibitor such as carfilzomib or oprosomib, a BCl-2 inhibitor such as venetoclax, an MCl-1 inhibitor such as AMG-176, a monoclonal antibody such as daratumumab, and an imide-based immunomodulatory drug (IMiD) such as thalidomide, lenalidomide, pomalidomide and apremilast.

In some embodiments, a compound having the structure

or a stereoisomer thereof, an atropisomer thereof, a pharmaceutically acceptable salt thereof, a pharmaceutically acceptable salt thereof of a stereoisomer, or a pharmaceutically acceptable salt thereof of an atropisomer, is administered to a patient in need thereof, a therapeutically effective amount of an additional pharmaceutically active compound, for example cytarabine, a proteasome inhibitor such as carfilzomib or oprosomib, a BCl-2 inhibitor such as venetoclax, an MCl-1 inhibitor such as AMG-176, a monoclonal antibody such as daratumumab, and an imide-based immunomodulatory drug (IMiD) such as thalidomide, lenalidomide, pomalidomide and apremilast.

In some embodiments, a compound having the structure

or a stereoisomer thereof, an atropisomer thereof, a pharmaceutically acceptable salt thereof, a pharmaceutically acceptable salt thereof of a stereoisomer, or a pharmaceutically acceptable salt thereof of an atropisomer, is administered to a patient in need thereof, a therapeutically effective amount of an additional pharmaceutically active compound, for example cytarabine, a proteasome inhibitor such as carfilzomib or oprosomib, a BCl-2 inhibitor such as venetoclax, an MCl-1 inhibitor such as AMG-176, a monoclonal antibody such as daratumumab, and an imide-based immunomodulatory drug (IMiD) such as thalidomide, lenalidomide, pomalidomide and apremilast.

In some embodiments, a compound having the structure

or a stereoisomer thereof, an atropisomer thereof, a pharmaceutically acceptable salt thereof, a pharmaceutically acceptable salt thereof of a stereoisomer, or a pharmaceutically acceptable salt thereof of an atropisomer, is administered to a patient in need thereof, a therapeutically effective amount of an additional pharmaceutically active compound, for example cytarabine, a proteasome inhibitor such as carfilzomib or oprosomib, a BCl-2 inhibitor such as venetoclax, an MCl-1 inhibitor such as AMG-176, a monoclonal antibody such as daratumumab, and an imide-based immunomodulatory drug (IMiD) such as thalidomide, lenalidomide, pomalidomide and apremilast.

In some embodiments, a compound having the structure

or a stereoisomer thereof, an atropisomer thereof, a pharmaceutically acceptable salt thereof, a pharmaceutically acceptable salt thereof of a stereoisomer, or a pharmaceutically acceptable salt thereof of an atropisomer, is administered to a patient in need thereof, a therapeutically effective amount of an additional pharmaceutically active compound, for example cytarabine, a proteasome inhibitor such as carfilzomib or oprosomib, a BCl-2 inhibitor such as venetoclax, an MCl-1 inhibitor such as AMG-176, a monoclonal antibody such as daratumumab, and an imide-based immunomodulatory drug (IMiD) such as thalidomide, lenalidomide, pomalidomide and apremilast.

In some embodiments, a compound having the structure

or a stereoisomer thereof, an atropisomer thereof, a pharmaceutically acceptable salt thereof, a pharmaceutically acceptable salt thereof of a stereoisomer, or a pharmaceutically acceptable salt thereof of an atropisomer, is administered to a patient in need thereof, a therapeutically effective amount of an additional pharmaceutically active compound, for example cytarabine, a proteasome inhibitor such as carfilzomib or oprosomib, a BCl-2 inhibitor such as venetoclax, an MCl-1 inhibitor such as AMG-176, a monoclonal antibody such as daratumumab, and an imide-based immunomodulatory drug (IMiD) such as thalidomide, lenalidomide, pomalidomide and apremilast.

The compounds described herein can be used in combination with agents that

- 48 044387 covered herein, or by other suitable means depending on the condition being treated. Therefore, in some embodiments, one or more compounds of the present invention will be administered in conjunction with other agents described above. When used in combination therapy, the compounds described herein are administered simultaneously or separately with a second agent. This combination administration may include simultaneous administration of two agents in the same dosage form, simultaneous administration in separate dosage forms, and separate administration. That is, the compound described herein and any of the agents described above can be formulated together in the same dosage form and administered simultaneously. Alternatively, the compound of the present invention and any of the agents described above can be administered simultaneously, with both agents present in separate formulations. In another alternative, administration of a compound of the present invention may be immediately followed by administration of any of the agents described above, or vice versa. In some embodiments of a separate administration protocol, a compound of the present invention and any of the agents described above are administered several minutes apart, or several hours apart, or several days apart.

Since one aspect of the present invention provides for the treatment of disease/conditions with a combination of pharmaceutically active compounds that can be administered separately, the present invention further relates to combining separate pharmaceutical compositions in the form of a kit. The kit contains two separate pharmaceutical compositions: a compound of the present invention and a second pharmaceutical compound. The kit contains a container for containing individual compositions, such as a divided bottle or a divided foil pouch. Additional examples of containers include syringes, boxes, and flexible reservoirs. In some embodiments, the kit contains instructions for use of the individual components. The kit form is particularly advantageous when the individual components are preferably administered in different dosage forms (eg, orally and parenterally), administered at different dosage intervals, or when dosing of the individual components of the combination will be directed by the health care professional who prescribes the treatment.

Examples

Method 1.

Example 1-1. 1 -(4-(6-(2-Bromo-5-hydroxyphenyl)-5-chloro-7-fluorobenzo[c]isothiazol-3-yl)piperazin-1 yl)prop-2-en-1-one

Step 1. 2-Amino-4-bromo-5-chloro-3-fluorobenzoic acid (intermediate A).

A mixture of 2-amino-4-bromo-3-fluorobenzoic acid (3.91 g, 16.71 mmol, Apollo Scientific Ltd., Stockport, UK) and N-chlorosuccinimide (1.36 ml, 16.7 mmol) in N ,N-dimethylformamide (33 ml) was stirred at 70°C for 20 hours. The reaction mixture was then cooled to room temperature, ice water (40 ml) was added and the resulting mixture was stirred for 1 hour. The resulting precipitate was collected by filtration , washed with water and dried under vacuum to obtain 2-amino-4-bromo-5-chloro-3-fluorobenzoic acid.

1H NMR (400 MHz, DMSO-d ₆ ) δ: 7.69 (1H, d, J=2.0 Hz), 6.48-7.23 (2H, br s).

^19F NMR (376 MHz, DMSO-d6) δ: -119.70 (1F, s).

Mass/charge (ESI, +ve) 270.0 (M+H) ⁺ .

Step 2: 2-Amino-4-bromo-5-chloro-3-fluorobenzamide (intermediate B).

Ammonium chloride (1.10 g, 20.6 mmol) and diisopropylethylamine (5.13 ml, 29.5 mmol) follower

- 49 044387 but added to a mixture of 2-amino-4-bromo-5-chloro-3-fluorobenzoic acid (intermediate A, 3.96 g, 14.7 mmol) and TBTU (4.97 g, 15.5 mmol , Advanced ChemTech, Louisville, KY, USA) in N,N-dimethylformamide (30 ml) and the resulting mixture was stirred at room temperature. within 30 min. The reaction mixture was then added to saturated aqueous sodium bicarbonate and stirred for 15 minutes. The resulting precipitate was collected by filtration, washed with water and dried under vacuum to obtain 2-amino-4-bromo-5-chloro-3-fluorobenzamide.

^1H NMR (400 MHz, DMSO-d6) δ: 8.03 (1H, br s), 7.72 (1H, d, J=2.0 Hz), 7.47 (1H, br s), 6 .86 (2H, s).

^19F NMR (376 MHz, DMSO-d6) δ: -120.79 (1F, s).

Mass/charge (ESI, +ve) 268.9 (m+H) ⁺ .

Step 3. 2-Amino-4-bromo-5-chloro-3-fluorobenzothioamide.

Lawsson's reagent (2.81 g, 6.95 mmol) was added to 2-amino-4-bromo-5-chloro-3-fluorobenzamide (intermediate B, 3.10 g, 11.59 mmol) in THF (77 ml ) and the resulting mixture was stirred at room temperature. for 1 hour. The reaction mixture was then diluted with EtOAc (75 ml) and washed successively with aqueous 2 M HCl (50 ml), saturated aqueous sodium bicarbonate (50 ml) and saline (50 ml). The organic extract was then dried over Na ₂ SO ₄ , collected by filtration and concentrated in vacuo. Chromatographic purification of the residue (silica gel, 0-3% MeOH in DCM) provided 2-amino-4-bromo-5-chloro-3-fluorobenzothioamide.

1H NMR (400 MHz, DMSO-d6) δ: 9.93-10.15 (1H, m), 9.63 (1H, br s), 7.28 (1H, d, J=1.96 Hz) , 6.34 (2H, s).

^19F NMR (376 MHz, DMSO-d6) δ: -119.52 (1F, s).

Mass/charge (ESI, +ve) 284.8 (m+H) ⁺ .

Step 4. 6-Bromo-5-chloro-7-fluorobenzo[c]isothiazol-3-amine.

Hydrogen peroxide (30% by weight in water, 2.93 mL, 28.7 mmol) was added dropwise to an ice-cold solution of 2-amino-4-bromo-5-chloro-3-fluorobenzothioamide (2.71 g, 9. 55 mmol) in pyridine (32 ml), and then the resulting mixture was heated to room temperature. and stirred it for 24 hours. Water (50 ml) was added and the precipitated solid was collected by filtration, washed with water and dried under vacuum to give 6-bromo-5-chloro-7-fluorobenzo[c]isothiazol-3- amine.

1H NMR (400 MHz, DMSO-d ₆ ) δ: 8.12-8.26 (2H, m), 7.95-8.06 (1H, m).

¹⁹ F NMR (376 MHz, DMSO-d ₆ ) δ: -114.32 (1F, s).

Mass/charge (ESI, +ve) 283.0 (M+H) ⁺ .

Step 5. 6-Bromo-3,5-dichloro-7-fluorobenzo[c]isothiazole (intermediate C).

To an ice-cooled mixture of 6-bromo-5-chloro-7-fluorobenzo[c]isothiazol-3-amine (2.47 g, 8.78 mmol), water (12 ml) and concentrated hydrochloric acid (37 wt.%, 12 ml, 395 mmol) a solution of sodium nitrite (0.788 g, 11.4 mmol) in water (2.0 ml) was slowly added. The resulting mixture was stirred at 0°C for 2.5 hours and then a mixture of copper chloride (1.39 g, 14.1 mmol) in concentrated hydrochloric acid (37 wt.%, 12 ml, 395 mmol) was added at 0° WITH. Then the reaction mixture was heated to room temperature. and it was stirred for 20 hours. The reaction mixture was diluted with water (50 ml) and the precipitated solid was collected by filtration and dried under vacuum. The collected material was taken up with (3:1) DCM:MeOH (200 ml) and washed successively with water (200 ml) and saline (100 ml). The organic layer was then dried over Na ₂ SO ₄ , filtered and concentrated in vacuo. Chromatographic purification of the residue (silica gel, 0-20% EtOAc in heptane) provided 6-bromo-3,5-dichloro-7-fluorobenzo[c]isothiazole.

1H NMR (400 MHz, DMSO-d ₆ ) δ: 7.99 (1H, d, J=1.57 Hz).

¹⁹ F NMR (376 MHz, DMSO-d ₆ ) δ: -111.48 (1F, s).

Mass/charge (ESI, +ve) 425.0 (M+H) ⁺ .

Step 6. tert-Butyl 4-(6-bromo-5-chloro-7-fluorobenzo[c]isothiazol-3-yl)piperazine-1-carboxylate (intermediate D).

Mixture of 6-bromo-3,5-dichloro-7-fluorobenzo[c]isothiazole (intermediate C, 150 mg, 0.497 mmol) and 1-Boc-piperazine (204 mg, 1.09 mmol) in N,N-dimethylformamide (2.0 ml) was stirred at room temperature. for 20 hours. The reaction mixture was then adsorbed onto silica gel and purified chromatographically (silica gel, 0-20% EtOAc in heptane) to give tert-butyl-4-(6-bromo-5-chloro-7fluorobenzo[c]isothiazol-3- yl)piperazine-1-carboxylate.

1H NMR (400 MHz, CHLOROFORM-d) δ: 7.60 (1H, d, J=1.56 Hz), 3.68-3.79 (4H, m), 3.40-3.51 (4H , m), 1.26 (9Н, s).

Mass/charge (ESI, +ve) 451.8 (M+H) ⁺ .

Step 7. tert-Butyl-4-(6-(2-bromo-5-methoxyphenyl)-5-chloro-7-fluorobenzo[c]isothiazol-3yl)piperazine-1-carboxylate.

Tert-Butyl 4-(6-bromo-5-chloro-7-fluorobenzo[c]isothiazol-3-yl)piperazine-1-carboxylate mixture (intermediate D, 111 mg, 0.247 mmol), 2-bromo-5 -methoxybenzeneboronic acid (0.114 ml, 0.494 mmol), sodium carbonate (0.041 ml, 0.988 mmol) and tetrakis-(triphenylphosphine) palladium (14.3 mg, 0.012 mmol) in 1,4-dioxane (1.6 ml) and water (0.4 ml) was heated at 90°C for 21 hours. Reaction

- 50 044387 the mixture was then concentrated in vacuo, adsorbed onto silica gel and purified by column chromatography (silica gel, 0-20% (3:1) EtOAc/EtOH in heptane) to give tert-butyl-4-(6-(2bromo- 5-methoxyphenyl)-5-chloro-7-fluorobenzo[c]isothiazol-3-yl)piperazine-1-carboxylate.

Mass/charge (ESI, +ve) 558.1 (M+H) ⁺ .

Step 8. 1-(4-(6-(2-Bromo-5-hydroxyphenyl)-5-chloro-7-fluorobenzo[c]isothiazol-3-yl)piperazin-1yl)prop-2-en-1-one .

Hydrogen chloride (4 M in 1,4-dioxane, 2.0 ml, 8.0 mmol) was added to a mixture of tert-butyl-4-(6-(2-bromo-5methoxyphenyl)-5-chloro-7-fluorobenzo[s ]isothiazol-3-yl)piperazine-1-carboxylate (107 mg, 0.192 mmol) and methanol (2.0 ml) and the resulting mixture was stirred at room temperature. for 1 hour. The reaction mixture was then concentrated in vacuo to give 6-(2-bromo-5-methoxyphenyl)-5-chloro-7-fluoro-3-(piperazin-1yl)benzo[c]isothiazole.

Mass/charge (ESI, +ve) 458.0 (M+1) ⁺ .

To this material (88 mg) was added N,N-diisopropylethylamine (0.101 ml, 0.578 mmol) in dichloromethane (2 ml) and the resulting mixture was cooled to 0°C. Acryloyl chloride (0.26 M in DCM, 0.75 mL, 0.19 mmol) was added and the resulting mixture was stirred at 0° C. for 10 minutes. The reaction mixture was concentrated in vacuo to give 1-(4-(6-(2-bromo-5-methoxyphenyl)-5-chloro-7fluorobenzo[c]isothiazol-3-yl)piperazin-1-yl)prop-2-ene -1-she.

Mass/charge (ESI, +ve) 512.0 (M+H) ⁺ .

For compounds without the methyl ether protecting group, the crude material was purified at this stage. For compounds bearing a methyl ether protecting group, the crude material was used in the next reaction without purification.

The resulting 1-(4-(6-(2-bromo-5-methoxyphenyl)-5-chloro-7-fluorobenzo[c]isothiazol-3-yl)piperazin-1yl)prop-2-en-1-one was absorbed with using 1,2-dichloroethane (2.0 ml) and cooled to 0°C. A solution of boron tribromide (1.0 M in hexanes, 0.97 mL, 0.97 mmol) was added and the resulting mixture was stirred at 0°C for 1 hour. The reaction mixture was then added to saturated aqueous sodium bicarbonate (2.0 mL ) and extracted with (2:1) DCM/MeOH (10 ml). The organic extract was dried over Na ₂ SO ₄ , filtered and concentrated in vacuo. Chromatographic purification of the residue (silica gel, 0-3% MeOH in DCM) provided 1-(4-(6-(2-bromo-5-hydroxyphenyl)-5-chloro-7fluorobenzo[c]isothiazol-3-yl)piperazin- 1-yl)prop-2-en-1-one.

¹ H NMR (400 MHz, DMSO-d6) δ: 9.99 (br s, 1H), 8.04 (s, 1H), 7.55 (d, J=8.7 Hz, 1H), 6, 81-6.94 (m, 2H), 6.79 (d, J=2.9 Hz, 1H), 6.19 (dd, J=16.7, 2.2 Hz, 1H), 5.77 (dd, J=10.5, 2.2 Hz, 1H), 3.87 (br d, J=19.5 Hz, 4H), 3.63 (br t, J=5.1 Hz, 4H) .

^19F NMR (376 MHz, DMSO-d6) δ: -124.16 (1F, s).

Mass/charge (ESI, +ve) 498.0 (m+H) ⁺ .

- 51 044387

Table 1(b)

Compounds 1-2 to 1-28 were prepared according to the procedure described in Method 1, Steps 1-8 above as follows

No. note Chemical structure Name Changes in the way Reagent 1-2 W ι, ^ι N ' LI g' „ У 1 NH 1-(4-(5-chloro-7- fluoro-6-(1Nindol-3-yl)-2,1benzothiazol-3yl)-1piperazinyl)-2propen-1-one stage 83 is excluded Step 7: (1-(tert-butoxycarbonyl)lh-indole-zyl)boronic acid (Combi-blocks Inc. San Diego, California, USA), stage 8-1: TFA/DCM 1-3 before Q sO tn - n; । OH 1-(4-(5-chloro-6- (2-fluoro-6hydroxyphenyl)2, l-benzothiazol3-yl)-lπipyrazinyl)-2propen-1-one stage 83 is excluded Stage 7: (3methoxynaphthal ey-1 yl)boronic acid, Cs ₂ CO ₃ , 100°C, stage 8-1: TFA/DCM 1-4 4 Q A'. OD 1 1 io uMe 1-(4-(5-chloro-6- (3-methoxy-1naphthalenyl)-2,1benzothiazol-3- il)-1- - Stage 7: (3methoxynaphthal ey-1 yl)boronic acid, Cs ₂ CO ₃ , 100°C, stage 8-1:

- 52 044387

piperazinyl)-2propen-1-one TFA/DCM 1-5 A Q 1l .ci 8% Ύ 'νΑΑΑ I j 1-(4-(5-chloro-6- (2-φτορ-6methoxyphenyl)2,1-benzothiazol3-yl)-1piperazinyl)-2propen-1-one - Step 7: (2-fluoro-6methoxyphenyl)boronic acid, Cs ₂ CO ₃ , 100°C, Step 8-1: TFA/DCM 1-6 J ¥4 4 ³ l 3^ -π 1-(4-(5-chloro-6- (2-fluoro-6hydroxyphenyl)2,1-benzothiazol3-yl)-1piperazinyl)-2propen-1-one - Step 7: (2-fluoro-6methoxyphenyl)boronic acid, Cs ₂ CO ₃ , 100°C, Step 8-1: TFA/DCM 1-7 ό ζ ο y44^ y C 44 l-((3R)-4-(5chloro-7-fluoro-6(3-hydroxy-1naphthalenyl)-2,1benzothiazol-3yl)-3(hydroxymethyl)1-piperazinyl)2-propen-1-one - Step 6: (3R)-1(tert-butoxycarbonyl)-3(hydroxymethyl)pipe razine (Synthonix Inc., Wake Forest, North Carolina, USA), step 7: (3methoxynaphthal en-1 yl)boronic acid (Ark Pharm Inc.,

-53 044387

Arlington Heights, Illinois, USA), Stage 8-1: TFA/DCM 1-8 -n. 1ί Ν 1 1 LI - F ΓΊ Ίι he ο Η - _Η -Ν Αν 4 ¹ F 1 1 I ¹ OH 1-4-(5-chloro-7- φτορ-6-(3- hydroxy-1naphthalenyl)-2,1benzothiazol-3yl)-3-ethyl-1piperazinyl)-2propen-1-one - Step 6: tert-butyl-3ethylpiperazine-1 carboxylate (Accel Pharmtech LLC, East Brunswick, NJ, USA), Step 7: (3methoxynaphthal eu-1 yl)boronic acid (Ark Pharm Inc., Arlington Heights, IL, USA) , stage 8-1: TFA/DCM 1-9 ,-r Η ' Γ ^Ν ._ ^! -ν L Γ 1 CI ·>' F / 1 ' ϊ ΌΗ Ji o' Π .1 1 ί ¹ Γ 7 ϊ - ΟΗ M-(1-(5-chloro-7fluoro-6-(3hydroxy-1naphthalenyl)-2,1benzothiazol-3yl)-2-methyl-3 azetidinyl)-2propenamide - Step 6: tert-butyl-(2methylazetidin-3yl)carbamate (Pharm aBlock, Nanjing, China), Step 7: (3methoxynaphthal ey-1yl)boronic acid (Ark Pharm Inc., Arlington Heights, IL, USA),

- 54 044387

Η A U i! n [1 Ί ( 1 , 1 ' J 1 Oh 1 ^G J- ΐ ¹ M 1 J ^cr I ' i Z and ' OH Stage 8-1: TFA/DCM 1-10 1 Q..... Ay A ^f Ju l-((3 _S )-4-(5chloro-7-fluoro-b(5-methyl-1Nindazol-4-yl)2,1-benzothiazol3 -yl)-3 -methyl-1 piperazinyl)-2propen-1- He stage 83 is excluded Step 6: (S)-4-nboc-2methylpiperazine (CNH Technologies, Inc., Woburn, MA, USA), Step 7: 4-boron-5-methyl- 1 h-indazole (Ark Pharm Inc., Arlington Heights, IL, USA), step 8-1: TFA/DCM 1-11 0\Vv ^c A ^h ° ^s uXX) p XjJ HO 1-((3 S )-4-(5chloro-7-fluoro-6(3-hydroxy-1naphthalenyl)-2,1benzothiazol-3- yl)-3-(2hydroxyethyl)-1 - - Step 6: (8)-tert-butyl-3-(2hydroxyethyl)piper zine-1-carboxylate (Activate Scientific GmbH, Prien, Germany), Step 7:

-55 044387

piperazinyl)-2propen-1-one (3-methoxynaphthal en1-yl)boronic acid (Ark Pharm Inc., Arlington Heights, IL, USA), step 8-1: TFA/DCM O Stage 6: piperazine- Η-Ν ^Ν , '' --Ν 4-(5-chloro-7- 2-carboxamide fi_“ /' Ν φτορ-6-(3- (Enamine, Kyiv, And 1 hydroxy-1- Ukraine), stage 7: Π I F L 1 11 - 'Ob naphthalenyl)-2,1- (3 -methoxynaphthalene- 1-12 benzothiazole-3- - 1-yl)boronic acid 'A. .if silt)-1-(2- acid (Ark Pharm Η Ν _ Ν propenoyl)-2- Inc., Arlington- ? Ν ΐι ι piperazinecarbox Heights, Illinois Li F amide USA), stage 8-1: I 1 < Η 1-(4-(5-chloro-7- TFA/DCM t fluorine-6-(2- Stage 7: (2- 1-13 γ /Л - Ν ι Ο ^S J Ί 1 ι ¹ methoxy-1naphthalenyl)-2,1benzothiazol-3- stage 8- is excluded methoxynaphthalen-1 yl)boronic acid, Cs ₂ CO ₃ , Υ ¹ il)-1- 3 100°C, stage 8-1: 1 piperazinyl)-2- TFA/DCM propen-1-one ¥ 1-(4-(5-chloro-7- Stage 7: (2- ργ fluorine-6-(2- methoxynaphthalene-1 - 1-14 4·ι -GI- hydroxy-1- - silt)boronic V-'J naphthalenyl)-2,1- acid, Cs ₂ CO ₃ , ^{FHO 1 1} benzothiazole-3- 100°C, stage 8-1:

- 56 044387

il)-1- piperazinyl)-2propen-1-one TFA/DCM 1-15 l/ Y T T G 1 F He 1-((3 S )-4-(5chloro-7-fluoro-b(3-hydroxy-1naphthalenyl)-2,1benzothiazol-3- yl)-3(hydroxymethyl)1-piperazinyl)2-propen-1-one - Step 6: (3S)-l-boc3-(hydroxymethyl)piperazine (Combiblocks Inc., San Diego, CA, USA), Step 7: (3methoxynaphthalen-1yl)boronic acid, Step 8-1: TFA/DCM 1-16 Λ- -Ν А ΐι' ι lj f i ] 1 ОН О hc ⁿ ' . π L. I 1 bl F * 1 1 > Ή 1-(4-(5-chloro-7- fluorine-6-(3- hydroxy-1naphthalenyl)-2,1benzothiazol-3- il)-2- (hydroxymethyl)1-piperazinyl)2-propen-1-one - Step 6: tert-butyl-3-(((tert-butyldimethylsilyl)o xy)methyl)piperazine -1-carboxylate, Step 7: (3methoxynaphthal en-1 yl)boronic acid, Step 8-1: TFA/DCM 1-17 O^- H H LI A. J ιI T OH l-((lR,5S)-3-(5chloro-7-fluoro-6(3-hydroxy-1naphthalenyl)-2,1benzothiazol-3yl)-3,8- - Step 6: 8-Boc-3,8diazabicycle o[3.2.1]octane (Chem-Impex International, Inc., Wood Dale, IL,

- 57 044387

diazabicyclo[3.2. 1 ]octan-8-yl)-2propen-1-one USA), step 7: (3methoxynaphthal ey-1 yl)boronic acid (Ark Pharm Inc., Arlington Heights, IL, USA), step 8-1: TFA/DCM 1-18 ' - ^N 'g ΪΙ Ν 1 ί lI F ί Μ he ο XI J /Ν ί U _f 'F ί ί I ¹ - he 1-(4-(5-chloro-7- fluorine-6-(3- hydroxy-1naphthalenyl)-2,1benzothiazol-3- yl)-3(hydroxymethyl)1-piperazinyl)2-propen-1-one - Step 6: 4-n-boc-2hydroxymethylpiper ZIN (AstaTech, Inc., Bristol, PA, USA), Step 7: (3methoxynaphthalen-1yl)boronic acid (Ark Pharm Inc., Arlington Heights, IL, USA), Stage 8-1: TFA/DCM 1-19 Α 'Λ . <1 „ ^S N ^J . ' , Ι , ΐ .J CH 1-((3 S )-4-(5chloro-7-fluoro-6(3-hydroxy-1naphthalenyl)-2,1benzothiazol-3yl)-3-methyl-1 piperazinyl)-2- propen-1-one — Step 6: (S)-4-nboc-2methylpiperazine (CNH Technologies, Inc., Woburn, MA, USA), Step 7: (3methoxynaphthalen-1yl)boronic acid (Ark Pharm Inc.,

- 58 044387

Arlington Heights, Illinois, USA), Stage 8-1: TFA/DCM 1-20 WITH HN '1 .Cl -r G J OH 1-(3 -((5-chl op-7φτορ-6-(3hydroxy-1naphthalenyl)-2,1benzothiazol-3yl)amino)-1azetidinyl)-2propen-1-one - Step 6: 1-boc-Zaminoazetidine (Alfa Aesar, Haver Hill, MA, USA), Step 7: (3methoxynaphthal en-1 yl)boronic acid (Ark Pharm Inc., Arlington Heights, IL, USA), Step 8- 1: TFA/DCM 1-21 X 0 Ax . ¹ Λ ¹ tu OH l-((3R)-4-(5chloro-7-fluoro-6(3-hydroxy-1naphthalenyl)-2,1benzothiazol-3yl)-3-methyl-1 piperazinyl)-2propen-1-one - Step 6: (R)-4-nboc-2-methylpiperazine (CNH Technologies, Inc., Woburn, MA, USA), Step 7: (3methoxynaphthal en-1yl)boronic acid (Ark Pharm Inc., Arlington Heights, Illinois, USA), stage 8-1: TFA/DCM

- 59 044387

1-22 d-C ¹ N ί . Cl. ¹ . ^: . tone N-((3R)-l-(5chloro-7-fluoro-6(3-hydroxy-1naphthalenyl)-2,1benzothiazol-3yl)-3pyrrolidinyl)-2propenamide Step 6: (3R)-(-)-3tert-butoxycarbonyl nopyrrolidine (Oakwood Products, Inc., Estill, SC, USA), Step 7: (3methoxynaphthalen-1yl)boronic acid (Ark Pharm Inc., Arlington Heights, IL , USA), Stage 8-1: TFA/DCM 1-23 smart \ V a .. J 1 ¹ ί “G % / Oh N-((3R)-l-(5chloro-7-fluoro-6(3-hydroxy-1naphthalenyl)-2,1benzothiazol-3yl)-3piperidinyl)-2propenamide Step 6: (R)-tert-butyl-piperidin-3ylcarbamate (Combi-blocks Inc., San Diego, CA, USA), Step 7: (3methoxynaphthal eu-1yl)boronic acid (Ark Pharm Inc., Arlington Heights, Illinois, USA), stage 8-1: TFA/DCM 1-28 A° C? SI Г νΑΑ^ΌΗ ^f _F XT 1-(4-(5-chloro-7fluoro-6-(2-fluoro- 5hydroxyphenyl)benzo[c]isothiazol3-yl)piperazin1-yl)prop-2-en1-one Step 7: (2-fluoro-5methoxyphenyl)boronic acid (Combiblocks Inc., San Diego, CA, USA), _K2CO3 , Pd( _dppf ) _Cl2 DCM, 100°C, Step 8-1: TFA/DCM

Method 2.

Example 2-1. 1-(4-(5-Chloro-6-(3-hydroxy-1-naphthalenyl)[1,2]thiazolo[3,4-b]pyridin-3-yl)-1 piperazinyl)-2-propene-1 -He

- 60 044387

Stage 1. 2-Amino-6-bromo-5-chloronicotinic acid.

N-Chlorosuccinimide (2.78 g, 20.8 mmol) was added to a solution of 2-amino-6-bromonicotinic acid (4.51 g, 20.8 mmol, Ark Pharm Inc., Arlington Heights, IL, USA) in DMF (75 ml) and the resulting mixture was heated at 70°C for 2.5 hours. Then the heating was stopped and stirring was continued for 16 hours. The reaction mixture was then poured into ice water. After the ice melted, the resulting slurry was filtered through a fritted glass funnel. The collected solids were air dried to obtain 2-amino-6-bromo-5-chloronicotinic acid.

1H NMR (400 MHz, DMSO-d ₆ ) δ: 8.05 (s, 1H), 7.64 (br. s, 2H).

Mass/charge (ESI, +ve) 250.9 (M+H) ⁺ .

Step 2. tert-Butyl 4-(2-amino-6-bromo-5-chloronicotinoyl)piperazine-1-carboxylate.

TBTU (1.93 g, 6.0 mmol) was added to a solution of 2-amino-6-bromo-5-chloronicotinic acid (1.12 g, 4.5 mmol) in DMF (14 mL). After 5 min, the reaction mixture was treated sequentially with 1-Boc-piperazine (912 mg, 4.9 mmol) and DIPEA (2.33 ml, 13.4 mmol). The resulting solution was stirred at room temperature. over 25 hours, saturated aqueous NaHCO ₃ solution (75 ml) was added and the resulting mixture was extracted with DCM. The organic layer was separated and washed successively with water (2x), dried over anhydrous sodium sulfate and concentrated in vacuo. Chromatographic purification of the residue (silica gel, 0 to 7% MeOH in DCM) provided tert-butyl 4-(2-amino-6-bromo-5-chloronicotinoyl)piperazine-1-carboxylate.

1H NMR (400 MHz, DMSO- _d6 ) δ: 7.58 (s, 1H), 6.66 (s, 2H), 3.33 (s, 8H), 1.40 (s, 9H).

Mass/charge (ESI, +ve) 419.0 (M+H) ⁺ .

Step 3. tert-Butyl 4-(2-amino-6-bromo-5-chloropyridine-3-carbonothioyl)piperazine-1-carboxylate.

Lawsson's reagent (353 mg, 0.87 mmol) was added to a solution of tert-butyl 4-(2-amino-6-bromo-5chloronicotinoyl)piperazine-1-carboxylate (610 mg, 1.45 mmol) in THF (7, 5 ml) and the resulting solution was stirred at 50°C for 2.5 hours. Then the reaction mixture was cooled to room temperature. and it was successively treated with water (10 ml) and aqueous 1 N. HCl solution (4 ml). The resulting mixture was extracted with EtOAc (2x) and the combined extracts were dried over anhydrous sodium sulfate, filtered and concentrated in vacuo. Chromatographic purification of the residue (silica gel, 0-6% MeOH in DCM) provided tert-butyl 4-(2-amino-6-bromo-5-chloropyridine-3carbonothioyl)piperazine-1-carboxylate.

1H NMR (400 MHz, DMSO-d ₆ ) δ: 7.47 (s, 1H), 6.58 (br. s, 2H), 4.30 (ddd, J=13.3, 6.3, 3 .3 Hz, 1H), 4.014.13 (m, 2H), 3.68-3.77 (m, 1H), 3.51-3.59 (m, 1H), 3.40-3.50 ( m, 3H), 1.41 (s, 9H).

Mass/charge (ESI, +ve) 434.9 (M+H) ⁺ .

Step 4. tert-Butyl 4-(5,6-dichloroisothiazolo[3,4-b]niridin-3-yl)piperazine-1-carboxylate.

NCS (116 mg, 0.87 mmol) was added to a solution of tert-butyl 4-(2-amino-6-bromo-5-chloropyridine-3carbonothioyl)piperazine-1-carboxylate (343 mg, 0.79 mmol) in THF (8 ml) and the resulting solution was stirred at room temperature. within 20 min. A mixture of water (10 ml) and 1 M aqueous sodium sulfite (5 ml) was then added and the resulting mixture was extracted with EtOAc (2x). The combined extracts were dried over anhydrous sodium sulfate, filtered and concentrated in vacuo. Chromatographic purification of the residue (silica gel, 0 to 4% MeOH in DCM) provided tertbutyl 4-(5,6-dichloroisothiazolo[3,4-b]pyridin-3-yl)piperazine-1-carboxylate.

^1H NMR (400 MHz, CHLOROFORM-d) δ: 8.10 (s, 1H), 3.69-3.80 (m, 4H), 3.50-3.57 (m, 4H), 1, 51 (s, 9H).

Mass/charge (ESI, +ve) 389.0 (M+H) ⁺ .

Step 5. tert-Butyl-4-(5-chloro-6-(3-methoxynaphthalen-1-yl)isothiazolo[3,4-b]pyridin-3-yl)piperazine-1-carboxylate.

Tert-Butyl 4-(5,6-dichloroisothiazolo[3,4-b]pyridin-3-yl)piperazine-1-carboxylate mixture (154 mg, 0.36 mmol), (3-methoxynaphthalen-1-yl) boronic acid (287 mg, 1.42 mmol) and cesium carbonate (463 mg, 1.42 mmol) in 1,4-dioxane (8 ml) and water (2 ml) were bubbled with argon before adding tetrakis-(triphenylphosphine) palladium ( 41 mg, 0.04 mmol). The reaction mixture was again bubbled with argon, then heated in a sealed tube at 100°C for 25 hours. After cooling to room temperature. the reaction mixture was diluted with brine (40 ml) and extracted with EtOAc (2x). The combined extracts were dried over sodium sulfate, filtered and concentrated in vacuo. Chromatographic purification of the residue (silica gel, 0-3.5% MeOH in DCM) provided tert-butyl-4-(5-chloro-6-(3-methoxynaphthalen-1-yl)isothiazolo[3,4-b]pyridin- 3-yl)piperazine-1carboxylate.

Mass/charge (ESI, +ve) 511.1 (M+H) ⁺ .

Step 6. 5-Chloro-6-(3-methoxynaphthalen-1-yl)-3-(piperazin-1-yl)isothiazolo[3,4-b]pyridine.

Trifluoroacetic acid (560 μL, 7.6 mmol) was added to the solution of tert-butyl-4-(5-chloro-6-(3methoxynaphthalen-1-yl)isothiazolo[3,4-b]pyridin-3-yl)piperazin- 1-carboxylate (155 mg, 0.30 mmol) in DCM (6 ml) and the resulting solution was stirred at RT. for 2.3 hours, then concentrated in vacuo. Chromatographic purification of the residue (silica gel, 0-25% MeOH in DCM) provided 5-chloro-6-(3-methoxynaphthalen-1-yl)-3-(piperazin-1-yl)isothiazolo[3,4-b]pyridine as TFA salt.

¹ H NMR (400 MHz, DMSO-d ₆ ) δ: 8.78 (s, 1H), 7.94 (d, J=8.2 Hz, 1H), 7.46-7.53 (m, 2H ), 7.31 (d, J=3.7 Hz, 2H), 7.19 (d, J=2.4 Hz, 1H), 3.95 (s, 3H), 3.76-3.83 (m, 4H), 3.35-3.43 (m, 4H).

Mass/charge (ESI, +ve) 411.0 (M+H) ⁺ .

Step 7. 1-(4-(5-Chloro-6-(3-methoxy-1-naphthalenyl)[1,2]thiazolo[3,4-b]pyridin-3-yl)-1piperazinyl)-2-propene -1-he.

To an ice-cooled suspension of 5-chloro-6-(3-methoxynaphthalen-1-yl)-3-(piperazin-1-yl)isothiazolo[3,4b]pyridine (TFA salt; 100 mg, 0.19 mmol) in DCM (5 ml) was sequentially added with DIPEA (100 µl, 0.57 mmol) and acryloyl chloride (23 µl, 0.29 mmol). The resulting solution was stirred at 0°C for 70 minutes and a saturated aqueous solution of NaHCO ₃ (15 ml) was added. The resulting mixture was extracted with DCM (3x) and the combined extracts were dried over sodium sulfate, filtered and concentrated in vacuo. Chromatographic purification of the residue (silica gel, 0 to 7% MeOH in DCM) provided 1-(4-(5-chloro-6-(3-methoxy-1-naphthalenyl)[1,2]thiazolo[3,4-b ]pyridin-3 yl)-1-piperazinyl)-2-propen-1-one.

¹ H NMR (400 MHz, DMSO-d ₆ ) δ: 8.73 (s, 1H), 7.93 (d, J=8.2 Hz, 1H), 7.45-7.54 (m, 2H ), 7.25-7.39 (m, 2H), 7.19 (d, J=2.5 Hz, 1H), 6.86 (dd, J=16.7, 10.3 Hz, 1H) , 6.19 (dd, J=16.7, 2.3 Hz, 1H), 5.77 (dd, J=10.5, 2.3 Hz, 1H), 3.94 (s, 3H), 3.81-3.94 (m, 4H), 3.69-3.76 (m, 4H).

Mass/charge (ESI, +ve) 465.0 (M+H) ⁺ .

Step 8. 1-(4-(5-Chloro-6-(3-hydroxy-1-naphthalenyl)[1,2]thiazolo[3,4-b]pyridin-3-yl)-1piperazinyl)-2-propene -1-he.

Boron tribromide (1.0 M in hexanes, 400 μL, 0.40 mmol) was added dropwise to an ice-cold solution of 1-(4-(5-chloro-6-(3-methoxynaphthalen-1-yl)isothiazolo[ 3,4-b]pyridin-3-yl)piperazin-1yl)prop-2-en-1-one (37.3 mg, 0.08 mmol) in 1,2-dichloroethane (4 ml) and the resulting mixture was stirred at 0°C for 2.3 hours. Then saturated aqueous NaHCO ₃ solution (5 ml) was added and the resulting mixture was extracted with (4:1) DCM:MeOH (2x). The combined extracts were dried over sodium sulfate, filtered and concentrated in vacuo. Chromatographic purification of the residue (silica gel, 0-6% MeOH in DCM) provided 1-(4-(5-chloro-6-(3-hydroxy-1-naphthalenyl)[1,2]thiazolo[3,4-b] pyridin-3-yl)-1-piperazinyl)-2-propen-1-one.

¹ H NMR (400 MHz, DMSO-d ₆ ) δ: 9.97 (br. s, 1H), 8.72 (s, 1H), 7.79 (d, J=8.6 Hz, 1H), 7.42 (t, J=7.1 Hz, 1H), 7.17-7.28 (m, 3H), 7.09 (d, J=2.1 Hz, 1H), 6.86 (dd , J=16.7, 10.5 Hz, 1H), 6.19 (dd, J=16.7, 2.3 Hz, 1H), 5.74-5.79 (m, 1H), 3, 81-3.95 (m, 4H), 3.68-3.76 (m, 4H).

Mass/charge (ESI, +ve) 451.0 (M+H) ⁺ .

- 62 044387

table 2

Compounds 2-2 to 2-6 were prepared according to the procedure described in Method 2, Steps 1-8 above as follows

No. note Chemical structure Name Changes in method Reagents 2-2 A T μ k / . %g A' ¹ A 1-(4-(5-chloro-6- (2-fluoro-6methoxyphenyl) [1, 2]thiazolo[3,4]pyridin-3yl)-3-methyl-1 piperazinyl)-2propen-1-one rule out stage 8 Step 2: 1-Boc-3methylpiperazine (Accela ChemBio Inc., San Diego, CA, USA), Step 4: Nbromosuccinimide, Step 5: 2-fluoro-6methoxyphenylboronic acid 2-3 Y o / But 1-(4-(5-chloro-6- (2-fluoro-6hydroxyphenyl)[1,2]thiazolo[3,4]pyridin-3yl)-3-methyl-1- - Step 2: 1-Boc-3methylpiperazine (Accela ChemBio Inc., San Diego, CA, USA), Step 4: N-

- 63 044387

¾ Y Ν YA ' L ^Ν ί 1 Η<_| - piperazinyl)-2propen-1-one Bromosuccinimide, Step 5: 2-fluoro-6methoxyphenylborones acid 2-4 Ts '-Ν _L ' 1 ^s ΤΙ 1 η '' Ί 1 1 1 % ρ kzV <Π Ύ ί χ 1-(4-(5-chloro-6(3-methoxy-1naphthalenyl) [1,2]t iazolo[3,4/>]pyridin-3yl)-3 -methyl-1 piperazinyl)-2propen-1-one rule out stage 8 Step 2: 1-Boc-3methylpiperazine (Accela ChemBio Inc., San Diego, CA, USA), Step 4: Nbromosuccinimide 2-5 γν ΑΑ _Ν 4γ^ΟΗ ο-^ ''Τ' 1-(4-(5-chloro-6(3-hydroxy-1naphthalenyl) [1,2]t iazolo[3,4]pyridin-3yl)-3-methyl-1 piperazinyl)-2propen-1-one - Step 2: 1-Boc-3methylpiperazine (Accela ChemBio Inc., San Diego, CA, USA), Step 4: Nbromosuccinimide 2-6 X _tsn 0 \=/ Α V Α^ρί 1-(4-(5-chloro-6(5-methyl-1-indazol-4yl)[1,2]thiazolo[3, 4-/>]pyridin-3- rule out stage 8 Step 2: 1-Boc-3methylpiperazine (Accela ChemBio Inc., San Diego, CA, USA),

- 64 044387

Ly A ChHs» ^Ν ΥΛρ/ yl)-3-methyl-1 piperazinyl)-2propen-1-one Step 4: Nbromosuccinimide, Step 5: 4-boron-5methyl-1h-indazole (Ark Pharm Inc., Arlington Heights, IL, USA) 2-7 o , ^Ν ,Γ- —Ν A ί> Ν } 1 Cl r' 1 r A ΌΗ Ο 11 Ν ί' 1 α 'Ύ τ 1' Μ '' ΟΗ ο Ν- / , -Ν [ Г ⁵ Μ f Τ ΟΙ 'y' Τ ΓΊ: k ' he 1-(4-(5-chloro-7- fluorine-6-(3- hydroxy-1naphthalenyl)-2,1benzothiazol-3yl)-3,5-dimethyl1-piperazinyl)2-propen-1-one - Step 1: 2-amino-4bromo-3fluorobenzoic acid (Apollo Scientific Ltd., Stockport, UK), Step 2: 1-Boc-3,5dimethylpiperazine (Combi-blocks Inc., San Diego, CA, USA), Step 7 : (3methoxynaphthal en-1 yl)boronic acid (Ark Pharm Inc., Arlington Heights, IL, USA), step 8-1: TFA/DCM 2-8 // \ / O D 1-(4-(5-chloro-7fluoro-6-(5methyl-1Nindazol-4-yl)2,1-benzothiazol3-yl)-3(difluoromethyl)- - Step 1: 2-amino-4bromo-3fluorobenzoic acid (Apollo Scientific Ltd., Stockport, UK),

- 65 044387

0 Λ ^F XN ' ^hs ' Г r ⁿ N'-^ . .] -X Г nh 'J 1-piperazinyl)2-propen-1-one step 2: tert-butyl(difluoromethyl)piper zin-1-carboxylate (Enamine, Kyiv, Ukraine), step 7: (5-methyl-1Nindazol-4yl)boronic acid (Combi-Blocks, Inc.), step 8-1: TFA/DCM 2-9 0. .'j Ί Ж %' F / s T v No. / 1 J \.JJ o, · ,N.. ¹ x A ^Γ 8··ν .n / F C0 1-(4-(5-chloro-7fluoro-6-(2-fluoro- 6hydroxyphenyl)2,1-benzothiazol3-yl)-3(difluoromethyl)1-piperazinyl)2-propen-1-one stage 8-3 was carried out before stages 8-2 Step 1: 2-amino-4bromo-3fluorobenzoic acid (Apollo Scientific Ltd., Stockport, UK), step 2: tert-butyl 3(difluoromethyl)piper zine-1-carboxylate (Enamine, Kiev, Ukraine), step 7: 2fluoro- 6-Methoxyphenylboronic acid (Accela ChemBio Inc., San Diego, CA, USA), step 8-1: TFA/DCM

- 66 044387

•ABOUT N A 1' 1 Gl, F ' ί Ίι ' ΌΗ 1-(4-(5-chloro-7fluoro-6-(3hydroxy-1naphthalenyl)-2,1- Step 1: 2-amino-4bromo-3fluorobenzoic acid (Apollo Scientific Ltd., Stockport, UK), Step 2: 1-Boc-Zisopropyl-piperazine 2-10 □ Ν - ,1 -Ν ί Ν and A 1 1 L ' ' Mon benzothiazol-3yl)-3-(2propanil)-1piperazinyl)-2propen-1-one (Ark Pharm Inc., Arlington Heights, IL, USA), Step 7: (3methoxynaphthal en-1 yl)boronic acid (Ark Pharm Inc., Arlington Heights, IL, USA), Stage 8-1: TFA/DCM

Method 3.

Example 3-1. 1-(4-(5-Chloro-7-fluoro-6-(3-hydroxynaphthalen-1-yl)benzo[c]isothiazol-3-yl)piperazin-1yl)prop-2-en-1-one

Step 1. 1-(4-(6-Bromo-5-chloro-7-fluorobenzo[c]isothiazol-3-yl)piperazin-1-yl)prop-2-en-1-one.

0.2 M acryloyl chloride in DCM (1.240 mL, 0.248 mmol) was added to an ice-cold solution of 6-bromo-5-chloro-7-fluoro-3-(piperazin-1-yl)benzo[c]isothiazole (intermediate D, 87 mg, 0.248 mmol) and N,N-diisopropylethylamine (0.129 ml, 0.744 mmol) in dichloromethane (2.3 ml) and the resulting mixture was stirred at 0°C for 10 minutes. The mixture was then concentrated in vacuo and the residue was sonicated in MeOH (2 ml). The suspended solid was collected by filtration, washed with MeOH and dried under vacuum to give 1-(4(6-bromo-5-chloro-7-fluorobenzo[c]isothiazol-3-yl)piperazin-1-yl)prop- 2-en-1 -ona.

1H NMR (400 MHz, DMSO de) δ: 8.13 (1H, d, J=1.56 Hz), 6.84 (1H, dd, J=10.47, 16.73 Hz), 6.17 (1H, dd, J=2.35, 16.63 Hz), 5.66-5.82 (1H, m), 3.73-3.93 (4H, m), 3.55-3.67 (4H, m).

^19F NMR (376 MHz, DMSO-d6) δ: -113.39 (s, 1F).

Mass/charge (ESI, +ve) 405.8 (M+H)+.

Step 2. 1-(4-(5-Chloro-7-fluoro-6-(3-methoxynaphthalen-1-yl)benzo[c]isothiazol-3-yl)piperazin-1yl)prop-2-en-1- he (intermediate E).

Mixture of 1-(4-(6-bromo-5-chloro-7-fluorobenzo[c]isothiazol-3-yl)piperazin-1-yl)prop-2-en-1-one (intermediate D, 79 mg, 0.20 mmol), (3-methoxynaphthalen-1-yl)boronic acid (47.3 mg, 0.234 mmol), tetrakis-(triphenylphosphine) palladium (22.5 mg, 0.020 mmol) and sodium carbonate (83 mg, 0 .78 mmol) in water (0.500 ml) and 1,4-dioxane (2.0 ml) was heated at 100°C for 16 hours. The reaction mixture was then adsorbed onto silica gel and purified by chromatography (silica gel, 0- 3% MeOH in

DCM) to give 1 -(4-(5-chloro-7-fluoro-6-(3-methoxynaphthalen-1 -yl)benzo[c]isothiazol-3 -yl)piperazin1 -yl)prop-2-en-1 -she.

Mass/charge (ESI, +ve) 482.0 (M+H)+.

Step 3. 1-(4-(5-Chloro-7-fluoro-6-(3-hydroxynaphthalen-1-yl)benzo[c]isothiazol-3-yl)piperazin-1yl)prop-2-en-1- He.

Boron tribromide (1.0 M in hexanes, 0.664 mL, 0.664 mmol) was added to an ice-cold solution of 1-(4-(5-chloro-7-fluoro-6-(3-methoxynaphthalen-1-yl)benzo[s] isothiazol-3-yl)piperazin-1-yl)prop-2-en1-one (64 mg, 0.13 mmol) in 1,2-dichloroethane (2.0 ml) and the resulting mixture was stirred at 0°C for 1 hour. The reaction mixture was then added to saturated aqueous sodium bicarbonate (2.0 ml) and the resulting mixture was extracted with (2:1) DCM:MeOH (10 ml). The organic extract was dried over Na ₂ SO ₄ , filtered and concentrated in vacuo. Chromatographic purification of the residue (silica gel, 0-3% MeOH in DCM) provided 1-(4-(5-chloro-7-fluoro-6-(3hydroxynaphthalen-1-yl)benzo[c]isothiazol-3-yl)piperazine -1 -yl)prop-2-en-1 -ona.

1H NMR (400 MHz, DMSO-d6) δ: 9.90-10.04 (1H, m), 8.10 (1H, s), 7.80 (1H, d, J=8.41 Hz), 7.43 (1H, ddd, J=1.96, 6.11, 8.17 Hz), 7.16-7.31 (3H, m), 7.07 (1H, d, J=2.35 Hz), 6.87 (1H, dd, J=10.47, 16.73 Hz), 6.19 (1H, dd, J=2.25, 16.73 Hz), 5.77 (1H, dd , J=2.25, 10.47 Hz), 3.88 (4H, br d, J=19.56 Hz), 3.61-3.72 (4H, m).

¹⁹ F NMR (376 MHz, DMSO-d ₆ ) δ: -123.78 (s, 1F).

Mass/charge (ESI, +ve) 468.0 (M+H)+.

Alternative synthesis of intermediate E

Intermediate C Intermediate E

-(4-(5-Chloro-7-fluoro-6-(3-methoxynaphthalen-1-yl)benzo[c]isothiazol-3-yl)piperazin-1-yl)prop-2-en1-one (intermediate E, alternative synthesis).

A solution of 1-( piperazin-1yl)prop-2-en-1-one-bis-(2,2,2-trifluoroacetate) (961 mg, 2.61 mmol, eNovation Chemicals LLC, Bridgewater, NJ, USA) in N,N -dimethylformamide (5.6 ml) and X,X-diisopropylethylamine (1.243 ml, 7.12 mmol). The resulting mixture was stirred at room temperature. for 1 hour and then heated at 50°C for 22 hours. After cooling to room temperature. the reaction mixture was added to ice water (10 ml) and the resulting precipitate was collected by filtration and washed with water. The collected solid was adsorbed onto silica gel and purified by chromatography (silica gel, 0-3% MeOH in DCM) to give 1-(4-(6-bromo-5-chloro-7-fluorobenzo[c]isothiazol-3-yl)piperazin- 1-yl)prop-2-en-1-one.

- 68 044387

Table 3

Compounds 3-2 to 3-24 were prepared according to the procedure described in method 3, steps 1-3 above as follows

3-2 o Sn NM 41 UgO “ O 8-(5-chloro-7-fluoro3-(4-(2-propenoyl)1-piperazinyl)-2,1benzothiazol-6-yl)2(1H)-quinolinone rule out stage 3 Step 2: 8(4,4,5,5tetramethyl1,3,2dioxaborolan2-yl)quinolin-2(1H)-one (Chem Shuttle, Hayward, CA, USA), S-Phos Pd G3, aq. K ₂ CO ₃ , 1,4-dioxane 3—3 0 o 1-(4-(5-chloro-7fluoro-6-(8isoquinolinyl)-2,1-benzothiazol-3-yl)1-piperazinyl)-2propen-1-one rule out stage 3 Step 2: 8boronoisoquinolin (Frontier Scientific, Inc., Logan, UT, USA) 3-4 0 <XXXJh T HD 5-(5-chloro-7-fluoro3-(4-(2-propenoyl)1-piperazinyl)-2,1benzothiazol-6-yl)2(1H)-quinolinone rule out stage 3 Step 2: 5(4,4,5,5tetramethyl1,3,2dioxaborolan2-yl)quinolin2(III)-one (Ark Pharm Inc., Arlington Heights, Illinois, USA)

- 69 044387

3-5 Ά° 0 Μ wW ^H F TM 1-(4-(5-chloro-7fluoro-6-(5-methyl1H-indazol-4-yl)2,1-benzothiazol-Zyl)-1piperazinyl)-2propen-1-one rule out stage 3 Step 2: 4borono-5-methyl1h-indazole (Ark Pharm Inc., Arlington Heights, IL, USA) 3-6 Μ ⁹ 0 Μ VvA S' 1 f :Г _; ι 1-(4-(5-chloro-7fluoro-6-(2-fluoro-6hydroxyphenyl)2,1-benzothiazol-Zyl)-1piperazinyl)-2propen-1-one - Step 2: 2fluoro-6-methoxyphenylboronic acid (Accela ChemBio Inc., San Diego, CA, USA) 3-7 V 0 Mm γι _F s '' ] | | Vd _F 1-(4-(5-chloro-6(2,4-difluorophenyl)7-fluoro-2,1benzothiazol-3-yl)1-piperazinyl)-2propen-1-one rule out stage 3 Step 2: (2,4difluorophenyl)boronic acid (Combi-blocks Inc., San Diego, CA, USA) 3-8 , T — _0 l7 / ’ ,- J = 1-(4-(5-chloro-7fluoro-6-(5hydroxy-2methylphenyl)-2,1benzothiazol-3-yl)1-piperazinyl)-2propen-1-one rule out stage 3 Step 2: (5hydroxy-2methylphenylboronic acid (Combi-blocks Inc., San Diego, CA, USA)

- 70 044387

3-9 N-- WITH? ΧΧΧθ. I X I 1 -(4-(5-chloro-6-(2chloro-5methoxyphenyl)-7fluoro-2,1benzothiazol-3-yl)1-piperazinyl)-2propen-1-one rule out stage 3 Step 2: 2chloro-5methoxyphenylboronic acid (Combi-blocks Inc., San Diego, CA, USA) 3-10 h 0 -'To He 1-(4-(5-chloro-6- (2,4-difluoro-5hydroxyphenyl)-7fluoro-2,1benzothiazol-3-yl)1-piperazinyl)-2propen-1-one - Step 2: 1boronic acid-2,4difluoro-5methoxy-benzene (Combi-blocks Inc., San Diego, CA, USA) 3-11 f p l N-. N g-ι A Al n 1 -(4-(5-chloro-6-(2chloro-5- hydroxyphenyl)-7fluoro-2,1- benzothiazol-3-yl)1-piperazinyl)-2propen-1-one - Step 2: 2chloro-5methoxyphenylboronic acid (Combi-blocks Inc., San Diego, CA, USA) 3-12 1 o h N- /\ '-N WITH! to D', ,' m Nil 1 -(4-(6-(5-amino2-methylphenyl)-5chloro-7-fluoro-2,1benzothiazol-3-yl)1-piperazinyl)-2propen-1-one rule out stage 3 Step 2: (5amino-2methylphenyl)boronic acid (Combi-blocks Inc., San Diego, CA, USA)

-71 044387

3-13 Q Vv ^c| s' ] T | j IJ 0 M-(3-(5-chloro-7φτορ-3-(4-(2propenoyl)-1piperazinyl)-2,1benzothiazol-6-yl)- methylphenyl)acetami d rule out stage 3 Step 2: [5(acetylamino)-2methylphenyl]boronic acid (Combi-blocks Inc., San Diego, California, USA) 3-14 ” G N - N ¹ Λ 1 NH ₂ 1 -(4-(6-(5-amino2-fluorophenyl)-5chloro-7-fluoro-2,1-benzothiazol-3-yl)1-piperazinyl)-2propen-1-one rule out stage 3 Step 2: 2fluoro-5aminophenylboronic acid (Combi-blocks Inc., San Diego, CA, USA) 3-15 A ^;< jHT( Tl 1 -(4-(6-(5-amino- 2,3-difluorophenyl)5-chloro-7-fluoro-2,1—benzothiazol-3-yl)1-piperazinyl)-2- propen-1-one rule out stage 3 Step 2: 3boron-4,5difluoroaniline (Combi-blocks Inc., San Diego, CA, USA) 3-16 0 % OH 1-(4-(5-chloro-6- (2,3-difluoro-5hydroxyphenyl)-7fluoro-2,1benzothiazol-3-yl)1-piperazinyl)-2propen-1-one - Step 2: 2,3difluoro-5methoxyphenylboronic acid (Combi-blocks Inc., San Diego, CA, USA)

- 72 044387

% ,ο 1-(4-(5-chloro-6(2,4-dichloro-5- Stage 2: (2,4dichloro-5- 3-17 And Cl :- HE hydroxyphenyl)-7fluoro-2,1benzothiazol-3-yl)1-piperazinyl)-2propen-1-one - methoxyphenyl) boronic acid (Combi-blocks Inc., San Diego, CA, USA) 3-18 Να Q U U δΎΎ Ai Ci He 1 -(4-(5-chloro-6-(2chloro-4-fluoro-5hydroxyphenyl)-7fluoro-2,1benzothiazol-3-yl)1-piperazinyl)-2propen-1-one - Step 2: (2chloro-4-fluoro-5methoxyphenyl)boronic acid (Combi-blocks Inc., San Diego, CA, USA) Stage 2: (5- 3-19 ur GA and ,Κ/Ν s )|AA F α xb ΝΗ ₂ 1 -(4-(6-(5-amino2-chlorophenyl)-5chloro-7-fluoro-2,1-benzothiazol-3-yl)1-piperazinyl)-2propen-1-one rule out stage 3 amino-2chlorophenyl)boronic acid, hydrochloride (Combi-blocks Inc., San Diego, CA, USA) Stage 2: (5- Du Λ Γη 1 -(4-(6-(5-amino- amino-2,4- 3-20 ci 2,4-dichlorophenyl)5-chloro-7-fluoro-2,1benzothiazol-3-yl)- - dichlorophenyl) boronic acid (Combi-blocks F L DN nh ₂ 1 -piperazinyl)-2- Inc., San Diego, propen-1-one California, USA)

- 73 044387

3-21 1-(4-(5-chloro-7- fluoro-6-(4-(2propanyl)-3pyridinyl)-2,1- benzothiazol-3-yl)1-piperazinyl)-2- propen-1-one rule out stage 3 Step 2: (4isopropylpyridinium-3yl)boronic acid (Combi-Phos Catalysts Inc., Trenton, NJ, USA) 4 s hr Cl ⁴ Stage 2: 2-(2,3- 3-22 No. N.Q. Ar ci Ay OH 1-(4-(5-chloro-6(2,3-dichloro-5hydroxyphenyl)-7fluoro-2,1benzothiazol-3-yl)1-piperazinyl)-2propen-1-one - dichloro-5methoxyphenyl)4,4,5,5tetramethyl1,3,2dioxaborolane (Anisyn Inc., Kalamazoo, MI, USA) 3-23 A/4 4 s ⁷ ^ '4 TU f | 1-(4-(5-chloro-7fluoro-6-(naphthalene- 1yl)benzo[c]isothiazo l-3-yl)piperazin1-yl)prop-2-en-1- He rule out stage 3 Stage 1-1: TFA/DCM, stage 2: (1naphthalyl)boronic acid, Cs ₂ CO ₃ , 80°C 3-24 E 0 -N S' | 7v 1-(4-(5-chloro-7- fluoro-6-(quinolin-8yl)benzo[c]isothiazo l-3-yl)piperazin1-yl)prop-2-en-1- He rule out stage 3 Step 1-1: TFA/DCM, Step 2: 8quinolineboronic acid (Frontier Scientific Inc., Logan, UT,

- 74 044387

USA), Cs ₂ CO ₃ , 80°С 3-25 0 A F XX MN _g 3-amino-5-(5-chloro7-fluoro-3-(4-(2propenoyl)-1piperazinyl)-2,1benzothiazol-6yl)benzonitrile rule out stage 3 Step 1-1: TFA/DCM, Step 2: (3amino-5cyanophenyl)boronic acid (Combi-blocks Inc., San Diego, CA, USA), S-Phos Pd G3,K ₂ CO ₃ , 100°C

Method 4.

Example 4-1. 1-(6-(5-Chloro-7-fluoro-6-(3-hydroxynaphthalen-1-yl)benzo[c]isothiazol-3-yl)-2,6diazaspiro[3.3]heptan-2-yl)prop- 2-en-1-on

Step 1. tert-Butyl-6-(6-bromo-5-chloro-7-fluorobenzo[c]isothiazol-3-yl)-2,6-diazaspiro[3.3]heptane-2carboxylate.

Mixture of 6-bromo-3,5-dichloro-7-fluorobenzo[c]isothiazole (intermediate C, 169 mg, 0.562 mmol) and 2-Boc-2,6-diazaspiro[3.3]heptane (212 mg, 1.07 mmol, AstaTech, Inc., Bristol, PA, USA) in DMF (3.5 ml) was stirred at room temperature. for 5 hours. Ice water (5 ml) was added and the resulting mixture was stirred for 15 minutes. The resulting precipitate was collected by filtration, washed with water and dried under vacuum to give tert-butyl-6-(6-bromo-5chloro-7-fluorobenzo[c]isothiazol-3-yl)-2,6-diazaspiro[3.3] heptane-2-carboxylate.

'I NMR (400 MHz, DMSO-d ₆ ) δ: 7.52-7.74 (1H, m), 4.55 (4H, s), 4.09 (4H, s), 1.38 (9H , s).

¹⁹ F NMR (376 MHz, DMSO-d ₆ ) δ: -113.55 (IF, s).

Mass/charge (ESI, +ve) 464.0 (M+l).

Step 2. 1-(6-(6-Bromo-5-chloro-7-fluorobenzo[c]isothiazol-3-yl)-2,6-diazaspiro[3.3]heptan-2-yl)prop2-en-1- He.

A solution of hydrogen chloride (4 M in 1,4-dioxane, 5.0 ml, 20 mmol) was added to tert-butyl-6-(6-bromo-5chloro-7-fluorobenzo[c]isothiazol-3-yl)-2, 6-diazaspiro[3.3]heptane-2-carboxylate (249 mg, 0.538 mmol) in methanol (10 ml) and the resulting mixture was stirred at room temperature. for 2 hours. The reaction mixture was then concentrated in vacuo to give 6-bromo-5-chloro-7-fluoro-3-(2,6-diazaspiro[3.3]heptan-2yl)benzo[c]isothiazole.

Mass/charge (ESI, +ve) 363.8 (M+l) ⁺ .

N,N-diisopropylethylamine (0.281 mL, 1.61 mmol) in dichloromethane (3.0 mL) was added to this material and the resulting mixture was cooled to 0°C. Acryloyl chloride (0.2 M in DCM, 2.69 mL, 0.538 mmol) was then added and the resulting mixture was stirred at 0° C. for 10 minutes. The reaction mixture was then concentrated in vacuo and the residue was purified by chromatography (silica gel, 0-10% (3:1)

-75044387

EtOAc/EtOH in DCM) to give 1-(6-(6-bromo-5-chloro-7-fluorobenzo[c]isothiazol-3-yl)-2,6diazaspiro[3.3]heptan-2-yl)prop-2 -en-1-ona.

1H NMR (400 MHz, DMSO-d6) δ: 7.65 (1H, d, J=1.4 Hz), 6.25-6.36 (1H, m), 6.10 (1H, dd, J =17.0, 2.3 Hz), 5.64-5.72 (1H, m), 4.58 (4H, s), 4.47 (2H, s), 4.18 (2H, s) .

^19F NMR (376 MHz, DMSO-d6) δ: -113.54 (1F, s).

Mass/charge (ESI, +ve) 418.0 (m+N) ⁺ .

Step 3. 1-(6-(5-Chloro-7-fluoro-6-(3-methoxynaphthalen-1-yl)benzo[c]isothiazol-3-yl)-2,6-diazaspiro[3.3]heptan-2 -yl)prop-2-en-1-one.

A mixture of 1-(6-(6-bromo-5-chloro-7-fluorobenzo[c]isothiazol-3-yl)-2,6-diazaspiro[3.3]heptan-2-yl)prop-2en-1-one ( 102 mg, 0.245 mmol), (3-methoxynaphthalen-1-yl)boronic acid (59.3 mg, 0.294 mmol), tetrakis-(triphenylphosphine) palladium (28.3 mg, 0.024 mmol) and sodium carbonate (104 mg, 0.979 mmol) in water (0.5 ml) and 1,4-dioxane (2.0 ml) was heated at 100°C for 1 hour. The reaction mixture was then adsorbed onto silica gel and purified by chromatography (silica gel, 0-5% MeOH in DCM). The purified material was sonicated in MeOH and the suspended solid was collected by filtration, washed with MeOH and dried under vacuum to give 1-(6-(5-chloro-7-fluoro-6-(3-methoxynaphthalen-1-yl) benzo[c]isothiazol-3-yl)-2,6-diazaspiro[3.3]heptan-2yl)prop-2-en-1-one.

^1H NMR (400 MHz, DMSO-d6) δ: 7.93 (1H, d, J=8.4 Hz), 7.67 (1H, s), 7.45-7.57 (2H, m) , 7.23-7.36 (2H, m), 7.16 (1H, d, J=2.5 Hz), 6.27-6.39 (1H, m), 6.11 (1H, dd , J=17.0, 2.2 Hz), 5.65-5.76 (1H, m), 4.58-4.67 (4H, m), 4.50 (2H, s), 4, 22 (2H, s), 3.93 (3H, s).

^19F NMR (376 MHz, DMSO-d6) δ: -123.88 (1F, s).

Mass/charge (ESI, +ve) 494.0 (m+H) ⁺ .

Step 4. 1-(6-(5-Chloro-7-fluoro-6-(3-hydroxynaphthalen-1-yl)benzo[c]isothiazol-3-yl)-2,6-diazaspiro[3.3]heptan-2 -yl)prop-2-en-1-one.

Boron tribromide (1.0 M in hexanes, 0.638 mL, 0.638 mmol) was added to ice-cold 1-(6-(5-chloro-7-fluoro-6-(3-methoxynaphthalen-1-yl)benzo[c]isothiazole -3-yl)-2,6-diazaspiro[3.3]heptan-2yl)prop-2-en-1-one (63 mg, 0.128 mmol) in 1,2-dichloroethane (2.0 ml) and the resulting mixture was stirred at 0°C for 2 hours. The reaction mixture was then added to saturated aqueous sodium bicarbonate (2.0 ml) and the resulting mixture was extracted with (2:1) DCM:MeOH (10 ml). The organic extract was dried over Na ₂ SO ₄ , filtered and concentrated in vacuo. Chromatographic purification of the residue (silica gel, 0-2% MeOH (using 2 M ammonia) in DCM) provided 1-(6-(5-chloro-7-fluoro-6-(3-hydroxynaphthalen-1-yl)benzo[ c]isothiazol-3-yl)-2,6-diazaspiro[3.3]heptan-2-yl)prop-2-en-1-one.

1H NMR (400 MHz, DMSO-d6) δ: 9.82-10.04 (1H, m), 7.79 (1H, d, J=8.2 Hz), 7.66 (1H, s), 7.43 (1H, dt, J=8.3, 4.0 Hz), 7.26 (1H, d, J=2.3 Hz), 7.22 (2H, d, J=3.7 Hz ), 7.05 (1H, d, J=2.3 Hz), 6.26-6.38 (1H, m), 6.12 (1H, dd, J=16.8, 2.2 Hz) , 5.66-5.72 (1H, m), 4.58-4.67 (4H, m), 4.50 (2H, s), 4.22 (2H, s).

^19F NMR (376 MHz, DMSO-d6) δ: -123.98 (1F, s).

Mass/charge (ESI, +ve) 480.0 (m+H) ⁺ .

- 76 044387

Table 4

Compounds 4-2 to 4-9 were prepared according to the procedure described in method 4, steps 1-4 above as follows

- 77 044387

ptan-2-yl)-2propen-1-one 4-4 0 ^Л-мн / / Ν - Cl - Ί ': ν - _r br 1 N-( 1 -(6-bromo-5chloro-7-fluoro- 2,1-benzothiazol3-i l )-3azetidinyl)-2propenamide exclude xia stage 3 u4 Step 1: 3-N-bocamino-azetidine, HCl salt (Combi-blocks Inc., San Diego, CA, USA) 4-5 o Κνη ~ δ π ^S JC1 χχ τ y N-( 1 -(5-chloro-7fluoro-6-(3methoxy-1naphthalenyl)-2,1benzothiazol-3yl)-3azetidinyl)-2propenamide excludes xia stage 4 Step 1: 3-N-bocamino-azetidine, HC1-salt (Combi-blocks Inc., San Diego, CA, USA) 4-6 0 b ΑΎ ΧΊ I Xp he N-( 1 -(5-chloro-7fluoro-6-(3hydroxy-1naphthalenyl)-2,1benzothiazol-3yl)-3azetidinyl)-2propenamide - Step 1: 3-N-bocamino-azetidine, HC1-salt (Combi-blocks Inc., San Diego, CA, USA) 4-7 0 TsCOan A' ¹ F 1-(3-((6-bromo-5chloro-7-fluoro- 2,1-benzothiazol3-yl)amino)-1azetidinyl)-2propen-1-one exclude xia stage 3 u4 Step 1: 1-boc-Zaminoazetidine (Alfa Aesar, Haver Hill, MA, USA)

- 78 044387

4-8 A (Ush Laf F l-((3R)-3-((6bromo-5-chloro-7fluoro-2,1benzothiazol-3yl)amino)-1piperidinyl)-2propen-1-one exclude xia stage 3 and 4 Step 1: (R)-Tperbutyl-3aminopiperidine 1-carboxylate (AstaTech, Inc., Bristol, PA, USA) 4-9 Ύ··° A t, _n jn / I 1 OH l-((3R)-3-((5chloro-7-fluoro-6(zhydroxynaphthale n-1yl)benzo[c]isothia zol-3yl)amino)piperidin-1-yl)prop2-en-1-one - Step 1: (R)-TpeTbutyl-3aminopiperidine 1-carboxylate (AstaTech, Inc., Bristol, PA, USA)

Method 5.

Example 5-1. N-(1 -(5-Chloro-7-fluoro-6-(3-hydroxy-1-naphthalenyl)-2,1-benzothiazol-3-yl)-3azetidinyl)-N-methyl-2-propenamide

Step 1. 2-Amino-5-chloro-3-fluoro-4-(3-methoxynaphthalen-1-yl)benzamide.

A mixture of (3-methoxynaphthalen-1-yl)boronic acid (2.04 g, 10.1 mmol), 2-amino-4-bromo-5-chloro-3fluorobenzamide (intermediate B (1.93 g, 7.20 mmol), tetrakis-(triphenylphosphine) palladium (0.832 g, 0.720 mmol), sodium carbonate (1.2 ml, 28.8 mmol) in water (9.6 ml) and 1,4-dioxane (38.4 ml) heated at 90°C for 2 days. The reaction mixture was then filtered through a pad of celite, washed with EtOAc. The filtrate was diluted with saturated aqueous NaHCO ₃ (50 ml) and extracted with EtOAc (3x50 ml). The organic extract was washed with saline ( 30 ml) and dried over Na ₂ SO _4. The solution was then filtered and the filtered material was concentrated in vacuo. The residue was suspended in MeOH (5 ml) and the suspended solid was collected by filtration, washed with MeOH and dried to obtain 2-amino-5 -chloro-3fluoro-4-(3-methoxynaphthalen-1-yl)benzamide Chromatographic purification of the concentrated filtrate (silica gel, 0% to 100% (3:1) EtOAc-EtOH in heptane) provided additional 2-amino-5 -chloro-3-fluoro-4-(3-methoxynaphthalen-1-yl)benzamide.

^1H NMR (400 MHz, DMSO-d6) δ: 8.01-8.17 (m, 1H), 7.92 (d, J=8.2 Hz, 1H), 7.75 (s, 1H) , 7.43-7.55 (m, 3H), 7.23-7.34 (m, 2H), 7.10 (d, J=2.5 Hz, 1H), 6.73 (s, 2H ), 3.93 (s, 3H).

Mass/charge (ESI, +ve) 345.0 (M+H) ⁺ .

Step 2. 2-Amino-5-chloro-3-fluoro-4-(3-methoxynaphthalen-1-yl)benzothioamide.

To a solution of 2-amino-5-chloro-3-fluoro-4-(3-methoxynaphthalen-1-yl)benzamide (2.11 g, 6.12 mmol) in tetrahydrofuran (41 ml) was added Lawsson's reagent (1.49 ml, 3.67 mmol) and the resulting mixture was stirred at room temperature. within 1 hour.

The reaction mixture was then diluted with EtOAc (60 ml) and washed successively with 2 M HCl (60 ml), saturated aqueous NaHCO ₃ solution (60 ml) and brine (60 ml). The organic extract was dried over Na ₂ SO ₄ , filtered and concentrated in vacuo. The residue was sonicated in DCM (5 ml) and the resulting precipitate was collected by filtration, washed with DCM and dried under vacuum to give 2-amino-5-chloro-3fluoro-4-(3-methoxynaphthalen-1-yl)benzothioamide. Chromatographic purification of the filtrate (silica gel, 0% to 100% (3:1) EtOAc-EtOH in heptane) provided additional 2-amino-5-chloro-3fluoro-4-(3-methoxynaphthalen-1-yl)benzothioamide.

Mass/charge (ESI, +ve) 361.0 (M+H) ⁺ .

Step 3. 5-Chloro-7-fluoro-6-(3-methoxynaphthalen-1-yl)benzo[c]isothiazol-3-amine.

A solution of hydrogen peroxide (30% in water, 2.2 mL, 21.3 mmol) was slowly added to the ice-cold solution of 2-amino-5-chloro-3-fluoro-4-(3-methoxynaphthalen-1-yl)benzothioamide ( 1.92 g, 5.33 mmol) in pyridine (18 ml). The resulting mixture was heated to room temperature. and it was stirred at room temperature. for 18 hours. The reaction mixture was then diluted with water (60 ml) and the resulting precipitate was collected by filtration, washed successively with water and MeOH and dried under vacuum to give 5-chloro-7-fluoro-6-(3-methoxynaphthalene-1 -yl)benzo[c]isothiazol-3-amine.

1H NMR (400 MHz, DMSO-d6) δ: 8.14 (s, 2H), 7.99-8.03 (m, 1H), 7.93 (d, J=8.3 Hz, 1H), 7.48-7.55 (m, 1H), 7.47 (d, J=2.3 Hz, 1H), 7.31 (d, J=3.9 Hz, 2H), 7.16 (d , J=2.5 Hz, 1H), 3.94 (s, 3H).

^19F NMR (376 MHz, DMSO-d6) δ: -124.71 (s, 1F).

Mass/charge (ESI, +ve) 359.0 (m+H) ⁺ .

Step 4. 3,5-Dichloro-7-fluoro-6-(3-methoxynaphthalen-1-yl)benzo[c]isothiazole.

5-Chloro-7-fluoro-6-(3-methoxynaphthalen-1-yl)benzo[c]isothiazol-3-amine (1.55 g, 4.31 mmol) was added in portions over 15 minutes. to a suspension of copper(II) chloride (0.870 g, 6.47 mmol) and tert-butylnitrite (0.77 ml, 6.47 mmol) in acetonitrile (43 ml) at 65°C. The resulting mixture was stirred at 65°C for 30 minutes and then cooled to ambient temperature and diluted with ice water (50 ml). The precipitated solid was collected by filtration, washed with water and dried under vacuum. The residue was sonicated in DCM (10 ml) and the suspended solid was collected by filtration, washed with DCM and dried under vacuum to release unreacted 5-chloro-7-fluoro-6-(3-methoxyaphthalen-1yl)benzo[s] isothiazol-3-amine. The filtrate was concentrated in vacuo to give 3,5-dichloro-7fluoro-6-(3-methoxynaphthalen-1-yl)benzo[c]isothiazole.

1H NMR (400 MHz, DMSO-d6) δ: 7.98 (s, 1H), 7.96 (d, J=8.2 Hz, 1H), 7.49-7.56 (m, 2H), 7.28-7.36 (m, 2H), 7.24-7.28 (m, 1H), 3.95 (s, 3H).

^19F NMR (376 MHz, DMSO-d6) δ: -122.17 (s, 1F).

Mass/charge (ESI, +ve) 378.0 (M+H) ⁺ .

Step 5. tert-Butyl-(1 -(5-chloro-7-fluoro-6-(3-methoxynaphthalen-1 -yl)benzo[c]isothiazol-3-yl)azetidin-3-yl)(methyl)carbamate.

A mixture of 3,5-dichloro-7-fluoro-6-(3-methoxynaphthalen-1-yl)benzo[c]isothiazole (100 mg, 0.264 mmol), DIPEA (0.14 ml, 0.793 mmol) and 3-Boc- 3-Methylaminoazatidine (0.098 ml, 0.529 mmol, Beta Pharma Scientific, Inc.) in DMF (1.3 ml) was stirred at room temperature. for 18 hours. Ice water (3 ml) was then added and the resulting mixture was stirred for 15 minutes. The precipitated solid was then collected by filtration, washed with water and dried under vacuum to give tert-butyl-(1-(5-chloro-7-fluoro-6-(3-methoxynaphthalen-1-yl)benzo[c]isothiazole -3-yl)azetidin-3yl)(methyl)carbamate.

Mass/charge (ESI, +ve) 528.0 (M+H) ⁺ .

Step 6. N-(1 -(5-Chloro-7-fluoro-6-(3-hydroxy-1-naphthalenyl)-2,1-benzothiazol-3-yl)-3-azetidinyl)N-methyl-2- propenamide

The title compound was prepared from tert-butyl-(1-(5-chloro-7-fluoro-6-(3methoxynaphthalen-1-yl)benzo[c]isothiazol-3-yl)azetidin-3-yl)(methyl) carbamate (131.1 mg, 0.248 mmol) in three steps according to the procedure outlined in Method 1, Step 8.

1H NMR (400 MHz, DMSO-d ₆ ) δ: 9.89-10.10 (m, 1H), 7.79 (d, J=8.4 Hz, 1H), 7.73 (s, 1H) , 7.43 (ddd,

J=8.2, 5.1, 2.9 Hz, 1H), 7.20-7.30 (m, 3H), 7.05 (d, J=2.2 Hz, 1H), 6.81 (dd, J=16.7, 10.5 Hz, 1H), 6.10-6.23 (m,

1H), 5.69-5.81 (m, 1H), 5.37-5.59 (m, 1H), 4.63-4.74 (m, 3H), 4.53-4.61 ( m, 1H), 3.14-3.23 (m, 3H).

¹⁹ F NMR (376 MHz, DMSO-d ₆ ) δ: -124.10 (s, 1F).

Mass/charge (ESI, +ve) 468.0 (M+H) ⁺ .

- 80 044387

Table 5

Compounds 5-2 to 5-9 were prepared according to the procedure described in method 5, steps 1-6 above as follows

No. note Chemical structure Name Reagent 5-2 ABOUT Anh X ' eleven That He A-(1 -(5-chloro-7-fluoro6-(3-hydroxy-1naphthalenyl)-2,1benzothiazol-3-yl)-3methyl-3-azetidinyl)2-propenamide Step 5: 3-(Vosamino)-3methylazetidine hydrochloride (Advanced ChemBlocks, Inc., Burlingame, CA, USA) 5-3 o ' -ΝΗ - _g ' PN 'Ν A ' m h Ί G I he N-( 1 -(5-chloro-7-fluoro6-(3-hydroxy-1naphthalenyl)-2,1benzothiazol-3-yl)-3(hydroxymethyl)-Zazetidinyl)-2propenamide Step 5: tert-butyl-3(hydroxymethyl)azet idin-3-ylcarbamate (Oakwood Products, Inc., Estill, South Carolina, USA)

- 81 044387

5-4 v .....Q of fl ί 1D He 1-((25)-4-(5-chloro-7fluoro-6-(3-hydroxy1-naphthalenyl)-2,1benzothiazol-3-yl)-2methyl-1piperazinyl)-2propen-1-one Step 5: (S)-l-Nboc-2methylpiperazine (Combi-blocks Inc., San Diego, CA, USA) 5-5 X .η JM Η XJ / ¹ , 1 ι τ he 1-((1A,5A)-6-(5-chloro7-fluoro-6-(3hydroxy-1naphthalenyl)-2,1benzothiazol-3-yl)2,6diazabicyclo[3.2.0]he ptan-2-yl)- 2-propen1-one Step 5: tert-butyl-2,6diazabicyclo[3.2.0]g eptane-2-carboxylate (eNovation Chemicals LLC, Bridgewater, NJ, USA) 5-6 V Ν Η to a. -Ci l to ¹ L ^and _t :Ι ί he 1-((15.5^)-6-(5-chlorop7-φτop-6-(3hydroxy-lnaphthalenyl)-2,1benzothiazol-3-yl)- 2.6- diazabicyclo[3.2.0]he ptan-2-yl)-2-propen1-one Step 5: tert-butyl-2,6diazabicyclo[3.2.0]g eptane-2-carboxylate (eNovation Chemicals LLC, Bridgewater, NJ, USA) 5-7 γ Ν — X/ v l. γι SP Ch N4A/W I (J He 1 -((27?)-4-(5-chloro-7fluoro-6-(3-hydroxy1-naphthalenyl)-2,1benzothiazol-3-yl)-2methyl-1piperazinyl)-2propen-1-one Step 5: (A)-l-boc2-methyl-piperazine (J&W Pharmlab, LLC, Levitown, PA, USA)

- 82 044387

5-8 J n '% . 1 A in 1 il ' 1 he A 17 t ' -! ¹ he 1 -(g/is-2-(5-chloro-7fluoro-6-(3-hydroxy1-naphthalenyl)-2,1benzothiazol-3-yl)- 2.6- diazabicyclo[3.2.0]he ptan-6-yl)-2-pr open1-one Step 5: 1-(2,6diazabicyclo[3.2.0]g eptan-6-yl)prop-2en-1-one (eNovation Chemicals LLC, Bridgewater, NJ, USA) 5-9 O tdkG / o ϊφ he 1-(3-((5-chloro-7fluoro-6-(3hydroxynaphthalene-1yl)benzo[c]isothiazol- 3yl)(methyl)amino)azeti din-1-yl)prop-2-en1-one Step 5: 1 azetidine carboxylic acid, 3(methyl amino)-, 1,1- dimethylethyl ester

Method 6.

Example 6-1. 1-(4-(6-(6-Amino-3-chloro-2-pyridinyl)-5-chloro-7-fluoro-2,1-benzothiazol-3-yl)-1 piperazinyl)-2-propene-1 -He

Stage 1. tert-Butyl-4-(5-chloro-7-fluoro-6-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzo[c]isothiazol-3 -yl)piperazine-1-carboxylate.

Tert-Butyl 4-(6-bromo-5-chloro-7-fluorobenzo[c]isothiazol-3-yl)piperazine-1-carboxylate mixture (intermediate D, 1.10 g, 2.45 mmol), bis -(pinacolato)diboron (1.86 g, 7.34 mmol), potassium acetate (0.61 ml, 9.8 mmol) and Pd(dppf)Cl ₂ DCM (0.537 g, 0.734 mmol) in 1.4- dioxane (12 ml) was heated at 100°C for 40 hours. The reaction mixture was then concentrated in vacuo and purified by chromatography (silica gel, 0% to 100% (3:1) EtOAc-EtOH in heptane) to give tert-butyl- 4-(5-chloro-7fluoro-6-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzo[c]isothiazol-3-yl)piperazine-1-carboxylate.

1H NMR (400 MHz, DMSO-d6) δ: 7.85 (s, 1H), 3.59 (br d, J=4.7 Hz, 4H), 3.44-3.54 (m, 4H) , 1.43 (s, 9H), 1.35 (s, 5H), 1.15 (s, 7H).

^19F NMR (376 MHz, DMSO-d6) δ: -125.11 (s, 1F).

Mass/charge (ESI, +ve) 498.0 (M+H) ⁺ .

Step 2. tert-Butyl-4-(6-(6-amino-3-chloropyridin-2-yl)-5-chloro-7-fluorobenzo[c]isothiazol-3yl)piperazine-1-carboxylate.

Mixture of tert-butyl-4-(5-chloro-7-fluoro-6-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzo[c]isothiazol-3-yl )piperazine-1-carboxylate (99.5 mg, 0.200 mmol), SPhos Pd G3 (17.3 mg, 0.020 mmol), 6-bromo-5-chloropyridine-2-amine (Combi-blocks Inc., San Diego , California, USA, 124 mg, 0.6 mmol), sodium carbonate (85 mg, 0.80 mmol) in water (0.25 ml) and 1,2-DCE (0.75 ml) heated at 50°C for 2 hours. The reaction mixture was concentrated in vacuo and purified by chromatography (silica gel, 0 to 100% (3:1) EtOAc-EtOH in heptane) to give tert-butyl-4-(6-(6-amino-3- chloropyridin-2-yl)-5chloro-7-fluorobenzo[c]isothiazol-3-yl)piperazine-1-carboxylate.

Mass/charge (ESI, +ve) 498.0 (M+H) ⁺ .

Step 3. 1-(4-(6-(6-Amino-3-chloro-2-pyridinyl)-5-chloro-7-fluoro-2,1-benzothiazol-3-yl)-1-83 044387 piperazinyl) -2-propen-1-one.

The title compound was prepared from tert-butyl 4-(6-(6-amino-3-chloropyridin-2-yl)-5chloro-7-fluorobenzo[c]isothiazol-3-yl)piperazine-1-carboxylate (31 .6 mg, 0.063 mmol) in two steps according to the procedure outlined in Method 1, Step 8.

Ή NMR (400 MHz, DMSO-d6) δ: 7.97-8.10 (m, 1H), 7.60 (d, J=8.9 Hz, 1H), 6.86 (dd, J=16 ,6, 10.6 Hz, 1H), 6.57 (d, J=8.9 Hz, 1H), 6.38 (s, 2H), 6.19 (dd, J=16.8, 2, 3 Hz, 1H), 5.71-5.84 (m, 1H), 3.86 (br d, J=19.9 Hz, 4H), 3.63 (br d, J=1.0 Hz, 4H).

^19F NMR (376 MHz, DMSO-d6) δ: -126.04 (s, 1F).

Mass/charge (ESI, +ve) 452.0 (M+H) ⁺ .

Table 6

Compound 6-2 was prepared according to the procedure described in method 6, steps 1-3, above as follows.

No. note Chemical structure Name Reagent 6-2 -- N A-\Ai 1 -(4-(5-chloro-6-(3 chloro-2-pyridinyl)-7fluoro-2,1-benzothiazol3 -yl)-1-piperazinyl)2-propen-1-one Step 2: 2-bromo-Zchloropyridine

Method 7.

Example 7-1. 1-((3R)-4-(5 -Chloro-7-fluoro-6-(3 -hydroxy-1 -naphthalenyl)-2, 1 -benzothiazol-3 -yl)-3 (difluoromethyl)-1 -piperazinyl) -2-propen-1-one/1 -((3,5)-4-(5-chloro-7-fluoro-6-(3-hydroxy-1-naphthalenyl)2,1-benzothiazol-3-yl) -3-(difluoromethyl)-1-piperazinyl)-2-propen-1-one

connection A

Step 1. 2-Amino-5-chloro-3-fluoro-4-(3-methoxynaphthalen-1-yl)benzoic acid.

Prepared from intermediate A using a procedure similar to that described in Method 1, Step 7.

Mass/charge (ESI, +ve) 346.0 (M+H) ⁺ .

Step 2. tert-Butyl 4-(2-amino-5-chloro-3-fluoro-4-(3-methoxynaphthalen-1-yl)benzoyl)-3(difluoromethyl)piperazine-1-carboxylate.

Mixture of 2-amino-5-chloro-3-fluoro-4-(3-methoxynaphthalen-1-yl)benzoic acid (0.150 g, 0.434 mmol), TBTU (0.188 g, 0.586 mmol), tert-butyl-3-( difluoromethyl)piperazine-1-carboxylate (0.123 g, 0.521 mmol) and DIPEA (0.23 ml, 1.302 mmol) in DMF (4 ml) were stirred at ambient temperature for 3 hours. The reaction mixture was then washed with saturated aqueous solution NaHCO ₃ and aqueous wash solution were extracted with EtOAc. The combined organic layers were dried over Na ₂ SO ₄ , filtered and concentrated in vacuo. Chromatographic purification of the residue (silica gel, 0-40% EtOAc/heptane) provided tert-butyl-4-(2amino-5-chloro-3-fluoro-4-(3-methoxynaphthalen-1-yl)benzoyl)-3-( difluoromethyl)piperazine-1-carboxylate.

Mass/charge (ESI, +ve) 586 (M+Na) ⁺ .

Step 3. tert-Butyl-4-(2-amino-5-chloro-3-fluoro-4-(3-methoxynaphthalen-1-yl)phenylcarbonothioyl)-3 (difluoromethyl)piperazine-1-carboxylate.

Lawsson's reagent (0.041 ml, 0.10 mmol) was added to the solution of tert-butyl-4-(2-amino-5-chloro-3fluoro-4-(3-methoxynaphthalen-1-yl)benzoyl)-3-(difluoromethyl) piperazine-1-carboxylate (0.095 g, 0.168 mmol) in THF (4 ml) and the resulting mixture was stirred at 50°C for 18 hours. The reaction

- 84 044387 the mixture was then concentrated in vacuo and purified by column chromatography (silica gel, 0-30% EtOAc/heptane) to give tert-butyl-4-(2-amino-5-chloro-3-fluoro-4-(3 -methoxynaphthalen-1yl)phenylcarbonothioyl)-3-(difluoromethyl)piperazine-1-carboxylate.

Mass/charge (ESI, +ve) 602.2 (M+Na) ⁺ .

Step 4. tert-Butyl-4-(5-chloro-7-fluoro-6-(3-methoxynaphthalen-1-yl)benzo[c]isothiazol-3-yl)-3(difluoromethyl)piperazine-1-carboxylate.

NBS (0.022 g, 0.17 mmol) was added to a solution of tert-butyl-4-(2-amino-5-chloro-3-fluoro-4-(3methoxynaphthalen-1-yl)phenylcarbonothioyl)-3-(difluoromethyl)piperazine -1-carboxylate in THF (7 ml) and the resulting mixture was stirred at ambient temperature for 15 minutes. The reaction mixture was diluted with water and washed with 10% sodium thiosulfate. The aqueous wash solution was extracted with EtOAc and the combined organic layers were then concentrated in vacuo to give tert-butyl-4-(5-chloro-7-fluoro-6-(3-methoxynaphthalen-1-yl)benzo[c]isothiazol-3yl )-3-(difluoromethyl)piperazine-1-carboxylate.

Mass/charge (ESI, +ve) 578.2 (M+H) ⁺ .

Step 5. 1-((3R)-4-(5-Chloro-7-fluoro-6-(3-hydroxy-1-naphthalenyl)-2,1-benzotuazol-3-yl)-3(difluoromethyl)-1 -piperazinyl)-2-propen-1-one.

1-((3,5)-4-(5-Chloro-7-fluoro-6-(3-hydroxy-1-naphthalenyl)-2,1-benzothiazol-3-yl)-3-(difluoromethyl)-1piperazinyl )-2-propen-1-one was prepared using a procedure similar to that described in Method 1, Step 8.

¹ H NMR (400 MHz, DMSO-d ₆ ) δ: 10.13 (br. s., 1H) 8.12 (d, J=2.2 Hz, 1H), 7.80 (d, J=8 ,2 Hz, 1H), 7.43 (br t, J=7.0 Hz, 1H), 7.20-7.30 (m, 3H), 7.08 (dd, J=5.8, 2 ,2 Hz, 1H) 6.78-6.91 (m, 1H) 6.27-6.70 (m, 1H) 6.20 (dd, J=16.6, 2.0 Hz, 1H) 5 .76-5.84 (m, 1H) 4.73-4.87 (m, 1H) 4.19-4.72 (m, 2H) 3.55-3.90 (m, 3H) 3.36 -3.47 (m, 1H).

Mass/charge (ESI, +ve) 518.0 (M+H) ⁺ .

Table 7

Compounds 7-2 and 7-3 were prepared according to the procedure described in method 7, steps 1-5, above as follows No. note Chemical structure Name Reagent 7-2 o ¹ -N ' /· N f J P “ ' - 'T [ Ί ' ii ' - · - he 1 -(4-(5-chloro-7-fluoro6-(3-hydroxy-1naphthalenyl)-2,1benzothiazol-3-yl)-3(fluoromethyl)-1piperazinyl)-2propen-1-one Step 2: 3-Fluoromethylpiperazine-1carboxylic acid tert-butyl ester (eNovation Chemicals LLC, Bridgewater, NJ, USA) o '-N T A ί . сг _Г f L Г I - - he 7-3 O.-, i--, ' ^N . ' Y LI - % ¹ ,y g ί I .( he \·ΐ/'i' G II methyl-1 -(5-chloro-7fluoro-6-(3-hydroxy1-naphthalenyl)-2,1benzothiazol-3-yl)-4(2-propenoyl)-2piperazinecarboxylate Step 2: 4boc-piperazine-2carboxylic acid methyl ester (Combi-blocks Inc., San Diego, CA, USA) f 1 t he Method 8.

Example 8-1. 6-Chloro-7-(2-fluoro-6-hydroxyphenyl)-1-(2-(2-propanyl)phenyl)-4-(4-(2-propenoyl)-1piperazinyl)-2(1H)-quinazolinone

- 85 044387

Stage 1. 4-Bromo-5-chloro-2-fluorobenzamide.

A mixture of 4-bromo-5-chloro-2-fluorobenzoic acid (23.3 g, 92 mmol) in thionyl chloride (67 ml, 0.92 mol) was stirred at 70°C using a reflux condenser for 1 hour. The reaction mixture was then concentrated in vacuo and the residue taken up in 1,4-dioxane (200 ml), treated with ammonium hydroxide (30% aqueous, 82 ml, 0.64 mol) and stirred at room temperature. within 15 min. The reaction mixture was concentrated in vacuo to give 4-bromo-5-chloro-2fluorobenzamide.

Mass/charge (ESI, +ve) 251.8 (M+H) ⁺ .

Step 2. 4-Bromo-5-chloro-2-fluoro-N-((2-isopropylphenyl)carbamoyl)benzamide.

A mixture of 4-bromo-5-chloro-2-fluorobenzamide (5.90 g, 23.4 mmol) and oxalyl chloride (1 M in DCM; 12.9 mL, 25.7 mmol) in DCE (100 mL) was stirred using reflux at 80°C for 1 hour. The reaction mixture was then cooled to room temperature. and 2-isopropylaniline (6.62 mL, 46.7 mmol) was added. The resulting mixture was stirred at room temperature. for 15 minutes, then cooled to 0°C. The precipitated solid was removed by filtration and the collected filtrate was concentrated in vacuo to give 4-bromo-5-chloro-2-fluoro-N-((2-isopropylphenyl)carbamoyl)benzamide.

1H NMR (400 MHz, DMSO-d6) δ: 11.06 (br. s., 1H), 10.31 (s, 1H), 7.97-8.05 (m, 2H), 7.82 ( d, J=7.2 Hz, 1H), 7.32-7.38 (m, 1H), 7.14-7.25 (m, 2H), 3.11 (spt, J=6.8 Hz , 1H), 1.24 (d, J=6.8 Hz, 6H).

¹⁹ F NMR (376 MHz, DMSO-d ₆ ) δ: -113.6 (s, 1F).

Mass/charge (ESI, +ve) 412.7 and 414.6 (M+H) ⁺ .

Step 3. 7-Bromo-6-chloro-1-(2-isopropylphenyl)quinazoline-2,4(1H,3H)-dione (intermediate F).

KHMDS (1 M in THF, 8.30 mL, 8.30 mmol) was added to a mixture of 4-bromo-5-chloro-2-fluoro-(2isopropylphenyl)carbamoyl)benzamide (1.56 g, 3.77 mmol) in THF (19 ml) at -20°C and ensured that the resulting mixture was heated to room temperature. for 1 hour. The reaction mixture was then diluted with EtOAc (150 ml) and washed with saturated aqueous ammonium chloride (2x100 ml). The organic layer was dried over Na ₂ SO ₄ , filtered and concentrated in vacuo. The residue was suspended in DCM (5 ml), sonicated, collected by filtration and dried under vacuum to give 7-bromo-6-chloro-1-(2-isopropylphenyl)quinazoline2,4(1H,3H)-dione.

1H NMR (400 MHz, CDCl ₃ ) δ: 9.43 (br. s., 1H), 8.29 (s, 1H), 7.55-7.59 (m, 2H), 7.39-7 .44 (m, 1H), 7.16 (d, J=7.8 Hz, 1H), 6.75 (s, 1H), 2.59-2.77 (m, 1H), 1.17- 1.24 (m, 3H), 1.11 (d, J=6.8 Hz, 3H).

Mass/charge (ESI, +ve) 392.9 and 395.0 (M+H) ⁺ .

Step 4. 6-Chloro-7-(2-fluoro-6-methoxyphenyl)-1-(2-isopropylphenyl)quinazoline-2,4(1H,3H)-dione.

Mixture of 7-bromo-6-chloro-1-(2-isopropylphenyl)quinazoline-2,4(1H,3H)-dione (intermediate F, 1.17 g, 2.96 mmol), (2-fluoro-6 -methoxyphenyl)boronic acid (2.02 g, 11.9 mmol), SPhos Pd G3 (0.128 g, 0.148 mmol) and potassium carbonate (2 M in water, 4.45 ml, 8.90 mmol) in DME (30 ml) was stirred at 85°C for 16 hours. The reaction mixture was then diluted with EtOAc (150 ml) and washed with saturated aqueous NaHCO ₃ solution (3x100 ml). The organic layer was dried over Na ₂ SO ₄ , filtered and concentrated in vacuo. Chromatographic purification of the residue (silica gel, 0-50% EtOAc in heptane) provided 6-chloro-7-(2-fluoro-6-methoxyphenyl)-1-(2isopropylphenyl)quinazoline-2,4(1H,3H)-dione .

1H NMR (400 MHz, DMSO-d ₆ ) δ: 11.90 (d, J=1.2 Hz, 1H), 8.11 (d, J=3.3 Hz, 1H), 7.53-7 .59 (m, 1H), 7.48 (tt, J=7.0, 2.2 Hz, 1H), 7.38-7.44 (m, 1H), 7.32-7.37 (m , 2H), 6.93 (dd, J=8.4, 4.3 Hz, 1H), 6.86 (t, J=8.7 Hz, 1H), 6.15 (s, 1H), 3 .66 (d, J=30 Hz, 3H), 2.73 (dq, J=14.2, 7.0 Hz, 1H), 1.11 (t, J=7.1 Hz, 3H), 1 .03 (dd, J=12.7, 6.8 Hz, 3H).

- 86 044387 ¹⁹ F NMR (376 MHz, DMSO-d6) δ: -113.8 (s, 1F) -115.2 (s, 1F).

Mass/charge (ESI, +ve) 439.1 (M+H) ⁺ .

Step 5. 4,6-Dichloro-7-(2-fluoro-6-methoxyphenyl)-1-(2-isopropylphenyl)quinazolin-2(1H)-one.

To a solution of 6-chloro-7-(2-fluoro-6-methoxyphenyl)-1-(2-isopropylphenyl)quinazoline-2,4(1H,3H)-dione (0.395 g, 0.900 mmol) and Et ₃ N ( 0.753 mL, 5.40 mmol) in acetonitrile (9 mL), phosphorus oxychloride (0.503 mL, 5.40 mmol) was added and the resulting solution was stirred at 80°C for 1.5 h. The reaction mixture was concentrated in vacuo to give 4. 6-dichloro-7-(2-fluoro-6-methoxyphenyl)-1-(2isopropylphenyl)quinazolin-2(1H)-one.

Mass/charge (ESI, +ve) 457.1 (M+H) ⁺ .

Alternative procedure for stage 5 (apply as indicated in the table below). To a stirred mixture of the product from step 4 (1.0 eq), triethylamine (18.0 eq) and 1H-benzo[d][1,2,3]triazole (12 eq) in acetonitrile (0.07 M ) phosphorus oxychloride (6.0 eq.) was added and the resulting reaction mixture was stirred at 80°C for 3.5 hours. The reaction mixture was then slowly poured into rapidly stirred water (100 ml) at 10°C. The aqueous suspension was stirred for 15 min before extraction with EtOAc (100 ml). The organic layer was washed with brine (100 ml), dried over MgSO ₄ , filtered and concentrated in vacuo to give the benzotriazole adduct intermediate, which was used directly in step 6.

Step 6. tert-Butyl-4-(6-chloro-7-(2-fluoro-6-methoxyphenyl)-1-(2-isopropylphenyl)-2-oxo-1,2dihydroquinazolin-4-yl)piperazin-1- carboxylate.

4,6-Dichloro-7-(2-fluoro-6-methoxyphenyl)-1-(2-isopropylphenyl)quinazolin-2(1H)-one solution (prepared according to method 8, step 5), tert-butyl piperazine -1-carboxylate (0.335 g, 1.80 mmol) and Et ₃ N (0.753 ml, 5.40 mmol) in DCE (9 ml) were stirred at 60°C for 20 minutes. The reaction mixture was diluted with EtOAc (100 ml) and washed with saturated aqueous NaHCO ₃ solution (3x75 ml). The organic layer was dried over Na ₂ SO ₄ and concentrated in vacuo. Chromatographic purification of the residue (silica gel, 0-60% (3:1) EtOAc-EtOH in heptane) provided tert-butyl-4-(6-chloro-7-(2-fluoro-6-methoxyphenyl)-1-(2-isopropylphenyl )-2-oxo-1,2-dihydroquinazolin-4yl)piperazine-1-carboxylate.

Mass/charge (ESI, +ve) 607.3 (M+H) ⁺ .

Note: If (S)-1-(3-methylpiperazin-1-yl)prop-2-en-1-one-2,2,2-trifluoroacetate was used, it was synthesized as follows.

(S)-1 -(3-Methylpiperazin-1 -yl)prop-2-ene-1-one-2,2,2-trifluoroacetate

Stage 6-a. (S)-tert-Butyl-4-acryloyl-2-methylpiperazine-1-carboxylate.

Acryloyl chloride (1.34 mL, 16.5 mmol) was added to a solution of (S)-1-boc-2-methyl-piperazine (3.00 g, 15.0 mmol, Boc Sciences, Shirley, NY) in THF (30.0 ml) at -10°C and the resulting mixture was stirred at -10°C for 5 minutes. Triethylamine (6.26 mL, 44.9 mmol) was then added slowly and the resulting mixture was stirred at -10°C for 15 minutes before warming to room temperature. The reaction mixture was partitioned between EtOAc and saturated aqueous NaHCO ₃ . The aqueous layer was extracted with EtOAc (3x) and the organic layers were then combined, dried over MgSO ₄ , filtered and concentrated in vacuo. Chromatographic purification of the residue (silica gel, 0-100% EtOAc in heptane) provided (S)-tert-butyl-4-acryloyl-2-methylpiperazine-1-carboxylate.

1H NMR (400 MHz, DMSO-d6) δ: 6.72-6.85 (m, 1H), 6.10-6.18 (m, 1H), 5.68-5.76 (m, 1H) , 4.08-4.32 (m, 2H), 3.68-4.03 (m, 2H), 2.86-3.14 (m, 2H), 2.66-2.80 (m, 1H), 1.38-1.43 (s, 9H), 0.96-1.04 (m, 3H).

Mass/charge (ESI, +ve) 277.3 (M+Na) ⁺ .

Stage 6-b. (S)-1 -(3-Methylpiperazin-1 -yl)prop-2-ene-1-one-2,2,2-trifluoroacetate.

Mixture of (S)-tert-butyl-4-acryloyl-2-methylpiperazine-1-carboxylate (3.21 g, 12.62 mmol) and TFA (4.7 mL, 63.1 mmol) in DCM (16 mL) stirred at room temperature. for 24 hours. The reaction mixture was then concentrated in vacuo to give (S)-1-(3-methylpiperazin-1-yl)prop-2-en-1-one-2,2,2-trifluoroacetate.

1H NMR (400 MHz, DMSO-d6) δ: 8.70-8.99 (m, 1H), 6.74-6.91 (m, 1H), 6.12-6.26 (m, 1H) , 5.70-5.84 (m, 1H), 4.25-4.44 (m, 1H), 4.07-4.25 (m, 1H), 3.49-3.53 (m, 1H), 3.22-3.32 (m, 2H), 2.92-3.08 (m, 2H), 1.14-1.29 (m, 3H).

Mass/charge (ESI, +ve) 155.1 (M+H) ⁺ .

Step 7. 6-Chloro-7-(2-fluoro-6-methoxyphenyl)-1-(2-isopropylphenyl)-4-(piperazin-1-yl)quinazolin2(1H)-one.

A solution of tert-butyl 4-(6-chloro-7-(2-fluoro-6-methoxyphenyl)-1-(2-isopropylphenyl)-2-oxo-1,2dihydroquinazolin-4-yl)piperazine-1-carboxylate ( 0.594 g, 0.978 mmol) in TFA (4 ml) stirred

- 87 044387 at ambient temperature for 30 minutes. The reaction mixture was concentrated in vacuo to give 6-chloro-7-(2-fluoro-6-methoxyphenyl)-1-(2-isopropylphenyl)-4-(piperazin-1yl)quinazolin-2(1H)-one.

Mass/charge (ESI, +ve) 507.2 (M+H) ⁺ .

Step 8. 4-(4-Acryloylpiperazin-1-yl)-6-chloro-7-(2-fluoro-6-methoxyphenyl)-1-(2-isopropylphenyl)_quinazolin-2(1H)-one.

To an ice-cooled solution of 6-chloro-7-(2-fluoro-6-methoxyphenyl)-1-(2-isopropylphenyl)-4(piperazin-1-yl)quinazolin-2(1H)-one and DIPEA (0.85 ml, 4.9 mmol) in DCM (10 ml) at 0°C was added acryloyl chloride (0.079 ml, 0.98 mmol) and the resulting mixture was stirred at 0°C for 30 min. The reaction mixture was then diluted with EtOAc (100 ml) and washed with saturated aqueous NaHCO3 (3x75 ml). The organic layer was dried over Na ₂ SO ₄ , decanted and concentrated in vacuo. Chromatographic purification of the residue (silica gel, 0-100% (3:1) EtOAc-EtOH in heptane) provided 4-(4-acryloylpiperazin-1-yl)-6-chloro-7-(2-fluoro-6methoxyphenyl)-1 -(2-isopropylphenyl)quinazolin-2(1H)-one.

1H NMR (400 MHz, CDCl ₃ ) δ: 7.86 (d, J=1.2 Hz, 1H), 7.41-7.54 (m, 2H), 7.29-7.37 (m, 2H), 7.14 (dt, J=7.8, 1.7 Hz, 1H), 6.70-6.79 (m, 2H), 6.58-6.68 (m, 1H), 6 .50 (d, J=7.4 Hz, 1H), 6.39 (dd, J=16.8, 1.8 Hz, 1H), 5.75-5.84 (m, 1H), 3, 79-4.06 (m, 8H), 3.75 (s, 2H), 3.66 (s, 1H), 2.69 (tt, J=13.4, 6.8 Hz, 1H), 1 .20-1.24 (m, 3H), 1.07 (dd, J=6.8, 3.9 Hz, 3H).

¹⁹ F NMR (377 MHz, CDCl·,) δ: -113.05 (s, 1F) -113.55 (s, 1F).

Mass/charge (ESI, +ve) 561.2 (M+H) ⁺ .

Step 9. 6-Chloro-7-(2-fluoro-6-hydroxyphenyl)-1-(2-(2-propanyl)phenyl)-4-(4-(2-propenoyl)-1piperazinyl)-2(1 H )-quinazolinone.

Br ₃ (1 M in DCE, 3.3 mL, 3.3 mmol) was added to an ice-cold solution of 4-(4-acryloylpiperazin-1-yl)-6-chloro-7-(2-fluoro-6-methoxyphenyl) -1-(2-isopropylphenyl)quinazolin2(1H)-one (0.372 g, 0.663 mmol) in DCE (1.7 ml) and the resulting mixture was stirred at 0° C. for 20 minutes, then warmed to room temperature. and it was stirred at room temperature. for 2 hours. A saturated aqueous solution of NaHCO3 was added to the reaction mixture, followed by the addition of EtOAc (150 ml). The organic layer was separated and washed with saturated aqueous NaHCO3 (3x100 ml). The organic layer was then dried over Na ₂ SO ₄ , filtered and concentrated in vacuo. Chromatographic purification of the residue (silica gel, 0-100% (3:1) EtOAc-EUN in heptane) provided 6-chloro-7-(2-fluoro-6-hydroxyphenyl)-1-(2-(2-propanyl)phenyl )-4-(4-(2-propenoyl)-1piperazinyl)-2(1H)-quinazolinone.

1H NMR (400 MHz, DMSO-d6) δ: 10.06 (br. d., J=15.1 Hz, 1H), 8.03 (d, J=1.2 Hz, 1H), 7.51 -7.56 (m, 1H), 7.45 (t, J=7.6 Hz, 1H), 7.33 (tdd, J=7.5, 7.5, 3.8, 1.4 Hz , 1H), 7.14-7.25 (m, 2H), 6.84 (dd, J=16.8, 10.4 Hz, 1H), 6.62-6.74 (m, 2H), 6.14-6.26 (m, 2H), 5.71-5.78 (m, 1H), 3.71-3.99 (m, 8H), 2.52-2.59 (m, 1H ), 1.02-1.12 (m, 6H). ^19F NMR (377 MHz, DMSO-d6), δ -113.6 (s, 1F), -114.8 (s, 1F).

Mass/charge (ESI, +ve) 547.1 (M+H) ⁺ .

- 88 044387

Table 8

Compounds 8-2 to 8-6 were prepared according to the procedure described in method 8, steps 1-9, above as follows

No. note Chemical structure Name Changes in method Original material Reagents 8-2 1 1 s I - T 1 II 1 'U' ¹ 6-chloro-7-(2fluoro-6hydroxyphenyl)1-(2-(2propanyl)phenyl)-4-(4-(2propenoyl)-1piperazinyl)pyr ido[2,3fpyrimidin- 2(1H)-on stages 7 and 8 are excluded 2,5,6trichloronic cotinic acid Stage 4: sodium carbonate Step 6: 1-(piperazin1-yl)prop-2en-1-one (eNovation Chemicals LLC, Bridgewater, New Jersey, USA) 8-3 II = A „ fill 1 1 - 6-chloro-7-(2fluoro-6hydroxyphenyl)4-((2X)-2-methyl4-(2-propenoyl)1 -piperazinyl)- 1-(2-(2propanyl)phenyl) -2(1H)- quinazolinone - 4-bromo5-chloro-2fluorobenzoic acid Step 6: (5) tert-butyl-3methylpiperazine n-1carboxylate (CNH Technologies, Inc., Woburn, MA)

- 89 044387

4° l/ 6-chloro-1-(2,6diethylphenyl)-7(2-fluoro-6hydroxyphenyl)4-((25)-2-methyl- except 2,5,6- Step 2: 2,6diethylaniline, Step 5: benzotriazole, Step 6: (5)1-(3methylpiperazi there are trichlorni 8-4 4-(2-propenoyl)- n-1-yl)prop- Alli stage 7 cotin 1- 2-en-1-on- 1 node and 8 acid 'st' piperazinyl)pyr 2,2,2- ido[2,3- trifluoroacetate (Dpyrimidine- (cm. 2(1H)-on note about stage 6 for synthesis) 6-chloro-1-(4- Stage 2: 4- cyclopropyl-3- cyclopropylp I ¹ r y pyridinyl)-7- 4-bromo- iridine-3- 1 -by (2-fluoro-6- amine (Combi- 5-chloro-2- hydroxyphenyl)- Phos Catalysts 8-5 C - GI — fluorobenzo ί 11 1 I 4-(4-(2- Inc., Trenton, 1 I 1 .1 propenoyl) -!- ynaya New Jersey, 1 1'4 acid Ν piperazinil)- USA), 1.4- 2(1H)- dioxane/water, quinazolinone 100°С 1' p 6-chloro-7-(2- Stage 4: With G 1 fluorine-6- except 2,5,6- carbonate X N hydroxyphenyl)- sodium there are trichlorni 8-6 Ν >- .* 4 4-((25)-2-methyl- stage 7 Stage 6: (5)- 1 1 1 i cotin С ^: ' N h ' _Ί ”ί I 1 .,η 4-(2-propenoyl)- and 8 acid 1-(3- 1 -piperazinyl)- methylpiperazi g U 1-(2-(2- n-1-yl)prop- propanyl)phenyl) 2-en-1-on- pyrido[2,3- 2,2,2- (Dpyrimidine- trifluoroacetate 2(1H)-on (cm. note about stage 6 for synthesis)

Method 9.

Example 9-1. 6-Chloro-7-(2,3-dichloro-5-hydroxyphenyl)-4-((2S)-2-methyl-4-(2-propenoyl)-1piperazinyl)-1-(2-(2-propanyl) phenyl)-2(1H)-quinazolinone

Step 1. 7-Bromo-4,6-dichloro-1-(2-isopropylphenyl)quinazolin-2(1H)-one.

To a mixture of 7-bromo-6-chloro-1-(2-isopropylphenyl)quinazoline-2,4(1H,3H)-dione (intermediate F, 470 mg, 1.194 mmol) and DIPEA (0.623 ml, 3.58 mmol ) in acetonitrile (11.4 ml) was added phosphorus oxychloride (0.915 ml, 5.97 mmol). The resulting mixture was heated at 80°C for 2 hours, then cooled to ambient temperature and concentrated in vacuo to give 7-bromo-4,6dichloro-1-(2-isopropylphenyl)quinazoline-2(1H) -she.

Mass/charge (ESI, +ve) 413.0 (M+H) ⁺ .

Step 2. (S)-4-(4-Acryloyl-2-methylpiperazin-1-yl)-7-bromo-6-chloro-1-(2-isopropylphenyl)quinazolin-2(1H)-one.

A mixture of 7-bromo-4,6-dichloro-1-(2-isopropylphenyl)quinazolin-2(1H)-one (492 mg, 1.19 mmol), (S)-4-N-boc-2-methylpiperazine ( 478 mg, 2.39 mmol) and DIPEA (0.623 ml, 3.58 mmol) in DMF (2.3 ml) were stirred at room temperature. within 10 min. Ice water (10 ml) was then added and the resulting mixture was stirred for 15 minutes. The precipitated solid was collected by filtration, washed with water and dried under vacuum to give (S)-tert-butyl-4-(7-bromo-6-chloro-1-(2isopropylphenyl)-2-oxo-1,2- dihydroquinazolin-4-yl)-3-methylpiperazine-1-carboxylate.

Mass/charge (ESI, +ve) 577.1 (M+H) ⁺ .

TFA (2.0 mL, 26.8 mmol) was added to a solution of (S)-tert-butyl-4-(7-bromo-6-chloro-1-(2isopropylphenyl)-2-oxo-1,2-dihydroquinazoline- 4-yl)-3-methylpiperazine-1-carboxylate (297 mg, 0.516 mmol) in DCM (2.0 ml) and the resulting mixture was stirred at RT. within 15 min. Concentration of the resulting mixture under vacuum provided the production of (S)-7-bromo-6-chloro-1-(2isopropylphenyl)-4-(2-methylpiperazin-1 -yl)quinazolin-2(1H)-one.

Mass/charge (ESI, +ve) 477.0 (M+H) ⁺ .

Acryloyl chloride (0.258 M in DCM, 4.0 mL, 1.031 mmol) was added to the ice-cooled (S)-7-bromo-6-chloro-1-(2-isopropylphenyl)-4-(2-methylpiperazin-1-yl) mixture )quinazolin-2(1H)-one and DIPEA (0.269 ml, 1.547 mmol) in DCM (2.0 ml) and the resulting mixture was stirred at 0°C for 20 minutes. Concentration in vacuo followed by chromatographic purification of the residue (silica gel, 0-100% (3:1) EtOAc-EtOH in heptane) provided (S)-4-(4-acryloyl-2-methylpiperazin-1-yl)7-bromo -6-chloro-1-(2-isopropylphenyl)quinazolin-2(1H)-one.

1H NMR (400 MHz, DMSO-d ₆ ) δ: 7.91-8.08 (m, 1H), 7.49-7.67 (m, 2H), 7.41 (br d, J=5, 8 Hz, 1H), 7.21 (br s, 1H), 6.76-6.98 (m, 1H), 6.52-6.67 (m, 1H), 6.09-6.29 ( m, 1H), 5.75 (br s, 1H), 4.61-4.96 (m, 1H), 4.234.48 (m, 1H), 3.93-4.21 (m, 2H), 3.50-3.77 (m, 1H), 3.33-3.49 (m, 1H), 3.23-3.28 (m, 1H), 2.94-3.24 (m, 1H ), 1.27 (br d, J=9.3 Hz, 6H), 1.09 (br s, 3H).

Mass/charge (ESI, +ve) 531.1 (M+H) ⁺ .

Step 3. (S)-4-(4-Acryloyl-2-methylpiperazin-1-yl)-6-chloro-7-(2,3-dichloro-5-methoxyphenyl)-1-(2isopropylphenyl)quinazoline-2( 1H)-on.

Mixture of (S)-4-(4-acryloyl-2-methylpiperazin-1-yl)-7-bromo-6-chloro-1-(2-isopropylphenyl)quinazolin2(1H)-one (120 mg, 0.226 mmol), 2-(2,3-dichloro-5-methoxyphenyl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (82 mg, 0.272 mmol), Na ₂ CO ₃ (96 mg, 0.906 mmol) and pd(PPh ₃ ) ₄ (26.2 mg, 0.023 mmol) in 1,4-dioxane (1.6 ml) and water (0.4 ml) were heated at 90°C for 17 hours. The reaction mixture was then concentrated under vacuum and purified by chromatography (silica gel, 0-100% (3:1) EtOAc-EtOH in heptane) to give (S)-4-(4-acryloyl-2-methylpiperazin-1-yl)-6-chloro-7 -(2,3-dichloro-5-methoxyphenyl)-1-(2isopropylphenyl)quinazolin-2(1H)-one.

Mass/charge (ESI, +ve) 627.0 (M+H) ⁺ .

Step 4. 6-Chloro-7-(2,3-dichloro-5-hydroxyphenyl)-4-((2S)-2-methyl-4-(2-propenoyl)-1-piperazinyl)1 -(2-( 2-propanyl)phenyl)-2(1H)-quinazolinone.

BBr ₃ (1Mb hexanes, 0.32 mL, 0.320 mmol) was added to the ice-cooled mixture of (S)-4-(4-acryloyl-2-methylpiperazin-1-yl)-6-chloro-7-(2,3- dichloro-5-methoxyphenyl)-1-(2-isopropylphenyl)quinazolin-2(1H)-one (40 mg, 0.064 mmol) and DCE (1.0 ml) and the resulting mixture was stirred at 0°C for 30 min. Saturated aq. NaHCO ₃ (2.0 mL) was added and the resulting mixture was extracted with (2:1) DCM/MeOH (5 mL).

The organic extract was dried over Na2SO4, filtered and concentrated in vacuo. Chromatographic purification of the residue (silica gel, 0-10% MeOH in DCM) provided 6-chloro-7-(2,3-dichloro-5-hydroxyphenyl)-4-((2S)-2-methyl-4-(2- propenoyl)-1-piperazinyl)-1-(2-(2propanyl)phenyl)-2(1H)-quinazolinone.

1H NMR (400 MHz, DMSO-d6) δ: 10.42 (br d, J=17.0 Hz, 1H), 7.86-8.11 (m, 1H), 7.50-7.63 ( m, 1H), 7.47 (bit, J=6.0 Hz, 1H), 7.36 (t, J=7.5 Hz, 1H), 7.15-7.26 (m, 1H), 7.05 (d, J=2.3 Hz, 1H), 6.78-6.96 (m, 1H), 6.446.58 (m, 1H), 6.11-6.29 (m, 2H) , 5.71-5.82 (m, 1H), 4.68-4.98 (m, 1H), 3.96-4.52 (m, 3H), 3.52-3.85 (m, 2H), 3.34-3.51 (m, 1H), 2.95-3.26 (m, 1H), 1.27-1.41 (m, 3H), 0.95-1.13 ( m, 6H).

Mass/charge (ESI, +ve) 611.0 (M+H) ⁺ .

- 91 044387

Table 9

Compounds 9-2 to 9-14 were prepared according to the procedure described in Method 9, Steps 1-4 above as follows

No. note Chemical structure Name Changes in the way Original material Reagent 9-2 V ^e N • GI A = f . ;> in, m '—' Ν 0 , 7-bromo-6-chloro4-((25)-2methyl-4-(2propenoyl)-1piperazinyl)-1 (2-(2propanyl)phenyl) -2(1H)- quinazolinone stages 3 and 4 are excluded 7-bromo-6chloro-1-(2isopropylphenyl)quinazo Lin- 2, 4(1N,ZN)- dione - 9-3 _Ν ( ) Cl CI ΓΧ - .ж ОуАжО Vn' νη ₂ ο \. 7-(5-amino-2chlorophenyl)-6chloro-4-((25)-2methyl-4-(2propenoyl)-1piperazinyl)-1 (2-(2propanyl)phenyl) -2(1H)- quinazolinone stage 4 is excluded 7-bromo-6chloro-1-(2isopropylphenyl)quinazo Lin- 2, 4(1N,ZN)- dione Step 3: (5amino-2chlorophenyl)b oronic acid hydrochloride (Combiblocks Inc., San Diego, CA, USA)

- 92 044387

9-4 And 0 “Huh? A, YES l. .He O' Ν Ιί A A? V s/ AT A. A<A^ an сг ν Ασ 1-(2-(2butanyl)phenyl)6-chloro-7-(3hydroxy-1naphthalenyl)-4- (4-(2propenoyl)-1piperazinyl)- 2(W)- quinazolinone SM was obtained in accordance with VII method 8, stages 1-3 7-bromo-1(2-(secbutyl)phenyl)-6-chloroquinazole in- 2, 4(1H,3N)dione Method 8, Step 2: (2secbutylphenyl)amine (Key Organics Inc., Bedford, MA, USA), Step 2: 1-Bospiperazine, Step 3: (3methoxynaphthalen-1yl)boronic acid (Ark Pharm Inc., Arlington Heights, Illinois, USA), SPhos PdG3, K ₂ CO ₃ , 1,4dioxane/water, 100°C

- 93 044387

9-5 J N f Ί n L ci Π 1 ί ° Il 1 , _? HO-N 3 -(6-chl op-7-(2φτορ-6hydroxyphenyl)2-oxo-4-(4-(2propenoyl)-1piperazinyl)1(2H)quinazolinyl)ben zonitrile SM was prepared according to method 8, steps 1-3 3-(7-bromo6-chloro-2,4dioxo-3,4dihydroquine azoline- 1(2H)yl)benzonitril Method 8, Step 2: 3—aminobenzone itrile (Frontier Scientific Services, Inc., Newark, DE, USA), Step 2: 1-hospiperazine, Step 3: 2-fluoro-6-hydroxyphenylboronic acid (Combiblocks Inc., San Diego, CA, USA ), SPhos PdG3, K ₂ CO ₃ , 1,4dioxane/water, 100°C

-94044387

9-6 11 o 1 Γ'ι /\ 1 · j 1 0 h ° Ν' 6-chloro-1-(3cyclopropyl-4pyridinyl)-7(2-fluoro-6hydroxyphenyl)- 4-(4-(2propenoyl)-1piperazinyl)- 2(1H)quinazolinone SM was obtained in accordance with VII method 8, stages 1-3 7-bromo-6chloro-1-(3cyclopropylpyridine- yl )quinazoli n— 2,4(1H,3H)dione Method 8, Step 2: 3cyclopropyl pyridine-4amine (Combi-Phos Catalysts Inc., Trenton, NJ, USA), Step 2:1-Bospiperazine, Step 3: 2fluoro-6hydroxyphenylboronic acid (Combiblocks Inc., San Diego , California, USA), SPhos PdG3, K ₂ CO ₃ , 1,4dioxane/water, 100°C

- 95 044387

9-7-2 9-7-1 I < j Λ ¹ ί ¹ h ' Λ L ' P Ί 1 ^Η0 ' Ν ' ^Ί Α _Ν. 1 1 X Ν Ά У к ϊ j ι 1 1' ^Ηί ' Ν 6-chloro-1-(3cyclopropyl-4pyridinyl)-7(2-φτορ-6hydroxyphenyl)- 4-((25)-2methyl-4-(2propenoyl)-1piperazinyl)- 2(1H)- quinazolinone [2nd eluting isomer] 6-chloro-1-(3cyclopropyl-4pyridinyl)-7(2-fluoro-6hydroxyphenyl)4-((25)-2methyl-4-(2- SM was prepared according to method 8, steps 1-3 SM was prepared according to method 8, step 7-bromo-6chloro-1-(3cyclopropylpyridine- yl )quinazoli n— 2,4(1H,3H)dione 7-bromo-6chloro-1-(3cyclopropylpyridine- 4yl)quinazoli n- Method 8, Step 2: 3cyclopropyl pyridine-4amine (Combi-Phos Catalysts Inc., Trenton, NJ, USA), Step 3: 2fluoro-6hydroxyphenyl boronic acid (Combiblocks Inc., San Diego, CA, USA), SPhos PdG3, K ₂ CO ₃ , 1,4dioxane/water, 100°C Method 8, step 2: 3cyclopropyl pyridine-4amine (Combi-Phos Catalysts Inc.,

- 96 044387

propenoyl)-1piperazinyl)2(1H)quinazolinone [1st eluting isomer] 1-3 2.4(1N,ZN)- dione Trenton, NJ, USA), step 3: 2fluoro-6hydroxyphenylboronic acid (Combiblocks Inc., San Diego, CA, USA), SPhos PdG3, _K2CO3 , _1,4dioxane /water, 100°C 9-9 V U rSrYV Λ D A A mn Ο^Ν-^Υρ/ About Ν 6-chloro-1-(3cyclopropyl-4pyridinyl)-7(5-methyl-1Nindazol-4-yl)4-((25)-2methyl-4-(2propenoyl)-1piperazinyl)2(1H)quinazolinone SM was obtained in accordance with VII method 8, stages 1-3, excluding stage 4 7-bromo-6chloro-1-(3cyclopropylpyridin4yl)quinazoli n— 2,4(1H,3H)dione Method 8, Step 2: 3cyclopropyl pyridine-4amine (Combi-Phos Catalysts Inc., Trenton, New Jersey, USA), step 3: 4boron-5methyl-lh-

- 97 044387

indazole (Ark-Pharm Inc.), SPhos PdG3, K ₂ CO ₃ , 1,4dioxane/water, 100°C 9-10 Μ ³ Ν............ν { > Cl Cl Cl h :· : y/y 6-chloro-7-(2,3dichlorophenyl)4-((25)-2methyl-4-(2propenoyl)-1piperazinyl)-1 (2-(2propanyl)phenyl) -2(1H)- quinazolinone stage 4 is excluded 7-bromo-6chloro-1-(2isopropylphenyl)quinazo Lin- 2, 4(1N,ZN)- dione Stage 3: 2,3dichlorobenzolboronic acid (Alfa Aesar, Haver Hill, MA, USA) 9-11 chu ( ) Cl Cl 6-chloro-7-(2chlorophenyl)-4((25)-2-methyl4-(2propenoyl)-1piperazinyl)-1 (2-(2propanyl)phenyl) -2(1H)- quinazolinone stage 4 is excluded 7-bromo-6chloro-1-(2isopropylphenyl)quinazo Lin- 2, 4(1N,ZN)- dione Step 3: 2chlorobenzene oronic acid (Alfa Aesar, Haver Hill, MA, USA)

- 98 044387

9-12 7' - z-^2 ex ij 7-(lHbenzotriazol-1yl)-6-chloro-1- (2,6diethylphenyl)-4((25)-2-methyl4-(2propenoyl)-1piperazinyl)pyrido[2,3b]pyrimidin- 2(1H)-on SM was obtained in accordance with VII method 8, stages 1-3. Stages 22 and 2-3 are excluded; compound was not isolated at stage 2-1 2,5,6trichloronic otinic acid Method 8, step 2: 2,6 diethylanyl and. Step 1: benzotriazole (see alternative conditions for method 8, step 5), step 2-1: (5)-1-(3methylpiperazin-1yl)prop-2- en-1-on- 2,2,2trifluoroacetate (see note about method 8, step 6, for synthesis) 9-13 V 0 _N W oALHuON Ai 6-chloro-7-(3hydroxy-1naphthalenyl)-1(2-(2propanyl)phenyl)-4-(4-(2propenoyl)-1piperazinyl)- 2(1H)- - 7-bromo-6chloro-1-(2isopropylphenyl)quinazo Lin- 2, 4(1H,3N)dione Step 1: benzotriazole (see alternative conditions for method 8, stage 5), stage 2-1: tertiary

- 99 044387

quinazolinone butylpiper zin-1carboxylate, stage 2-3: NEt ₃ stage 3: SPhos Pd G3, (zmetoxinapht alen-1yl)boronic acid, DME 9-14 >—and Z—4 and -H ) - - - - “ v О 6-chloro-1-((17?)2,2- dimethylcyclohexyl)-7-(2fluoro-6hydroxyphenyl)4-(4-(2propenoyl)-1piperazinyl)pyr ido[2,3b]pyrimidin2(1H)-onbchloro-1-((15)2,2dimethylcyclohexyl)-7- (2fluoro-6hydroxyphenyl)4-(4-(2- SM was obtained in accordance with the II method 8, stages 1-3. Stages 22, 2-3 and 4 are excluded 2,5,6trichloronic otinic acid Method 8, step 2: 2,2dimethylcyclohexane-1amine, step 2(1): 1(piperazin1-yl)prop2-en-1-one (eNovation Chemicals LLC, Bridgewater, NJ, USA), step 3: propenoyl)-1piperazinyl)pyrido[2,3b]pyrimidine- 2(1H)-on SPhos Pd G3, 2-fluoro-6hydroxyphenylboronic acid (Combiblocks Inc., San Diego, California, USA), DME

Method 10.

Example 10-1. 1 -(4-(7-Chloro-6-(2-fluoro-6-hydroxyphenyl)-4-(2-methylphenyl)-1-phthalazinyl)-1 piperazinyl)-2-propen-1-one

- 100 044387

Step 1: 6,7-Dichloro-2,3-dihydrophthalazine-1,4-dione (intermediate G).

Hydrazine (0.232 ml, 10.1 mmol) was added to a mixture of 5,6-dichloroisobenzofuran-1,3-dione (2.00 g, 9.22 mmol, TCI America, Portland, OR, USA) and ethanol (30 ml) and the resulting mixture was refluxed for 2 hours before cooling to room temperature. The resulting precipitate was collected by filtration and washed with water to obtain 6,7-dichloro-2,3-dihydrophthalazine-1,4 dione.

Mass/charge (ESI, +ve) 231.1 (M+H) ⁺ .

Step 2. 6-Chloro-7-(2-fluoro-6-hydroxyphenyl)-2,3-dihydrophthalazine-1,4-dione.

Mixture of 6,7-dichloro-2,3-dihydrophthalazine-1,4-dione (intermediate G, 3.80 g, 16.45 mmol), 2-fluoro-6-hydroxyphenylboronic acid (10.26 g, 65. 8 mmol, Combi-blocks Inc., San Diego, CA, USA), SPhos Pd G3 (1.423 g, 1.645 mmol) and 2 M aqueous Na ₂ CO ₃ (32.9 ml, 65.8 mmol) in DME (60 ml) was stirred at 80°C for 16 hours. The reaction mixture was cooled to room temperature. and diluted with water (200 ml) and EtOAc (300 ml). The aqueous layer was separated and acidified with 5N. HCl and extracted with EtOAc (300 ml). The combined organic layers were washed with brine (200 ml), dried over MgSO ₄ , filtered and concentrated in vacuo. The residue was suspended in DCM (50 ml) and collected by filtration to give 6-chloro-7-(2-fluoro-6-hydroxyphenyl)-2,3-dihydrophthalazine-1,4-dione.

Mass/charge (ESI, +ve) 307.0 (M+H) ⁺ .

Step 3. 6-(2-((tert-Butyldiphenylsilyl)oxy)-6-fluorophenyl)-7-chloro-2,3-dihydrophthalazine-1,4dione.

tert-Butyl(chloro)diphenylsilane (2.67 mL, 10.25 mmol) was added to the ice-cooled mixture of 6-chloro-7-(2-fluoro-6-hydroxyphenyl)-2,3-dihydrophthalazine-1,4-dione (2.62 g, 8.54 mmol) and TEA (4.75 ml, 34.2 mmol) in acetonitrile (40 ml) and the resulting mixture was stirred at 0°C for 15 min, then warmed to room temperature. and stirred for 1.5 hours. Additional tert-butyl(chloro)diphenylsilane (2.67 mL, 10.25 mmol) was added and the resulting mixture was stirred at room temperature. for 16 hours. The reaction mixture was successively diluted with water (300 ml), acidified with 5N. HCl and extracted with EtOAc (300 ml). The organic layer was separated and washed successively with brine (250 ml), dried over MgSO ₄ , filtered and concentrated in vacuo. The residue was taken up in DCM (200 ml), TFA (20 ml) was added and the resulting mixture was stirred at room temperature. within 45 min. The reaction mixture was then diluted with saturated aqueous NaHCO3 (200 ml) and extracted with DCM (2x250 ml). The combined organic extracts were dried over MgSO ₄ , filtered and concentrated in vacuo to give 6-(2-((tert-butyldiphenylsilyl)oxy)-6-fluorophenyl)-7-chloro-2,3-dihydrophthalazine-1,4-dione.

Mass/charge (ESI, +ve) 545.2 (M+H)+.

- 101 044387

Step 4. 6-(2-((tert-Butyldiphenylsilyl)oxy)-6-fluorophenyl)-1,4,7-trichlorophthalazine.

Pyridine (1.45 ml, 17.1 mmol) was added to a mixture of 6-(2-((tert-butyldiphenylsilyl)oxy)-6fluorophenyl)-7-chloro-2,3-dihydrophthalazine-1,4-dione (4, 66 g, 8.55 mmol) and phosphorus oxychloride (6.39 ml, 68.4 mmol) and the resulting mixture was heated at 100°C for 1.5 hours. The reaction mixture was then cooled to room temperature. and slowly poured into stirred water (300 ml) while maintaining the internal temperature at <10°C. After stirring for 15 minutes, the resulting mixture was extracted with EtOAc (400 ml) and the organic extract was washed successively with brine (250 ml), dried over MgSO ₄ , filtered and concentrated in vacuo. Chromatographic purification of the residue (silica gel, 0-25% EtOAc in heptane) provided 6-(2-((tert-butyldiphenylsilyl)oxy)-6-fluorophenyl)-1,4,7-trichlorophthalazine.

Mass/charge (ESI, +ve) 581.1 (M+H) ⁺ .

Step 5. tert-Butyl 4-(6-(2-((tert-butyldiphenylsilyl)oxy)-6-fluorophenyl)-4,7-dichlorophthalazin-1yl)piperazine-1-carboxylate (intermediate H).

1-Boc-piperazine (5.00 g, 26.9 mmol) was added to a mixture of 6-(2-((tert-butyldiphenylsilyl)oxy)-6fluorophenyl)-1,4,7-trichlorophthalazine (5.21 g, 8 .95 mmol) and triethylamine (3.77 ml, 26.9 mmol) in DCM (35 ml) and the resulting mixture was stirred at room temperature. for 19 hours. The reaction mixture was then partitioned between DCM (300 ml) and saturated aqueous NaHCO ₃ solution (200 ml). The organic layer was separated, dried over MgSO ₄ , filtered and concentrated in vacuo. Chromatographic purification of the residue (silica gel, 0-50% EtOAc in heptane) provided a mixture of tert-butyl-4-(6-(2-((tert-butyldiphenylsilyl)oxy)-6-fluorophenyl)-4,7-dichlorophthalazin-1-yl )piperazine-1-carboxylate and tert-butyl 4-(7-(2-((tert-butyldiphenylsilyl)oxy)-6-fluorophenyl)-4,6-dichlorophthalazin-1-yl)piperazine-1-carboxylate. Individual regioisomers were isolated by chiral SFC purification (OJ-H column (30x250 mm, 5 µm), 15% (20 mM NH3 in MeOH) in supercritical CO ₂ ) to give tert-butyl-4-(6(2-(( tert-butyldiphenylsilyl)oxy)-6-fluorophenyl)-4,7-dichlorophthalazin-1-yl)piperazine-1-carboxylate as the second eluting isomer.

1H NMR (400 MHz, CDCl ₃ ) δ: 8.27 (s, 1H), 8.17 (s, 1H), 7.56-7.61 (m, 4H), 7.40-7.46 ( m, 2H), 7.31-7.37 (m, 4H). 6.99-7.07 (m, 1H), 6.77 (t, J=8.61 Hz, 1H), 6.42 (d, J=8.22 Hz, 1H), 3.72-3 .77 (m, 4H), 3.53-3.59 (m, 4H), 1.51 (s, 9H) 0.66 (s, 9H).

Mass/charge (ESI, +ve), 731.2 (M+H) ⁺ .

Step 6. 6-(2-((tert-Butyldiphenylsilyl)oxy)-6-fluorophenyl)-4,7-dichloro-1-(piperazin-1yl)phthalazine.

Trifluoroacetic acid (2 mL, 26.8 mmol) was added to the stirred solution of tert-butyl4-(6-(2-((tert-butyldiphenylsilyl)oxy)-6-fluorophenyl)-4,7-dichlorophthalazin-1-yl)piperazine -1-carboxylate (intermediate H, 1.21 g, 1.654 mmol) in DCM (10 ml) and the resulting mixture was stirred at RT. for 1.5 hours. The reaction mixture was then diluted with saturated aqueous NaHCO ₃ (75 ml) and extracted with DCM (2x100 ml). The combined organic extracts were dried over MgSO4, filtered and concentrated in vacuo to give 6-(2-((tert-butyldiphenylsilyl)oxy)-6-fluorophenyl)-4,7-dichloro-1-(piperazin-1-yl)phthalazine.

Mass/charge (ESI, +ve) 631.3 (M+H) ⁺ .

Step 7. 1-(4-(6-(2-((tert-Butyldiphenylsilyl)oxy)-6-fluorophenyl)-4,7-dichlorophthalazin-1yl)piperazin-1-yl)prop-2-en-1- He.

Acryloyl chloride (0.148 ml, 1.81 mmol) was added to a mixture of 6-(2-((tert-butyldiphenylsilyl)oxy)-6fluorophenyl)-4,7-dichloro-1-(piperazin-1-yl)phthalazine (1.04 g, 1.647 mmol) and triethylamine (0.694 ml, 4.94 mmol) in DCM (10 ml) and the resulting mixture was stirred at room temperature. within 45 min. Saturated aqueous NaHCO ₃ solution (75 ml) was added and the resulting mixture was extracted with DCM (3x100 ml). The combined organic extracts were dried over MgSO ₄ , filtered and concentrated in vacuo to give 1-(4-(6-(2-((tert-butyldiphenylsilyl)oxy)-6-fluorophenyl)-4,7dichlorophthalazin-1-yl)piperazin- 1-yl)prop-2-en-1-one.

Mass/charge (ESI, +ve) 685.1 (M+H) ⁺ .

Step 8. 1-(4-(4,7-Dichloro-6-(2-fluoro-6-hydroxyphenyl)phthalazin-1-yl)piperazin-1-yl)prop-2-en-1one (intermediate I) .

TBAF (1 M in THF, 3.3 mL, 3.30 mmol) was added to a solution of 1-(4-(6-(2-((tert-butyldiphenylsilyl)oxy)-6-fluorophenyl)-4,7-dichlorophthalazine-1 -yl)piperazin-1-yl)prop-2-en-1-one (1.13 g, 1.648 mmol) in THF (10 ml) and the resulting mixture was stirred at room temperature. within 15 min.

The reaction mixture was concentrated in vacuo and the residue was purified by column chromatography (silica gel, 0-100% EtOAc in heptane) to give 1-(4-(4,7-dichloro-6-(2-fluoro-6hydroxyphenyl)phthalazin-1-yl )piperazin-1-yl)prop-2-en-1-one.

1H NMR (400 MHz, DMSO-d6) δ: 10.26 (br s, 1H), 8.31 (s, 1H), 8.14 (s, 1H), 7.31-7.40 (m, 1H), 6.78-6.92 (m, 3H), 6.17 (dd, J=16.63, 2.35 Hz, 1H), 5.74 (dd, J=10.37, 2, 35 Hz, 1H), 3.79-3.92 (m, 4H), 3.46-3.55 (m, 4H).

Mass/charge (ESI, +ve) 447.0 (M+H) ⁺ .

Step 9. 1-(4-(7-Chloro-6-(2-fluoro-6-hydroxyphenyl)-4-(o-tolyl)phthalazin-1-yl)piperazine-1

- 102 044387 yl)prop-2-en-1-one.

A mixture of 1-(4-(4,7-dichloro-6-(2-fluoro-6-hydroxyphenyl)phthalazin-1-yl)piperazin-1-yl)prop-2-en-1-one (intermediate I, 25 mg, 0.056 mmol), 2-tolylboronic acid (30.4 mg, 0.224 mmol, Frontier Scientific Inc., Logan, UT, USA), Pd(PPh ₃ ) ₄ (6.46 mg, 5.59 μmol, Strem Chemicals Inc., Newburyport, MA, USA) and a 2 M aqueous solution of Na ₂ CO ₃ (0.084 ml, 0.168 mmol) in 1,4-dioxane (0.3 ml) was stirred at 40°C for 18 hours. The reaction mixture then diluted with EtOAc (20 ml) and washed with water (15 ml). The organic layer was separated and washed successively with brine (15 ml), dried over MgSO ₄ , filtered and concentrated in vacuo. Chromatographic purification of the residue (silica gel, 0-100% EtOAc in heptane) provided 1-(4-(7-chloro-6-(2-fluoro-6-hydroxyphenyl)-4-(o-tolyl)phthalazin-1-yl )piperazin-1-yl)prop-2en-1-one.

1H NMR (400 MHz, DMSO-d6) δ: 10.15 (br s, 1H), 8.33 (s, 1H), 7.36-7.45 (m, 2H), 7.24-7, 36 (m, 4H), 6.90 (dd, J=16.63, 10.37 Hz, 1H), 6.70-6.80 (m, 2H), 6.18 (dd, J=16, 73, 2.25 Hz, 1H), 5.75 (dd, J=10.56, 2.15 Hz, 1H), 3.83-3.97 (m, 4H), 3.47-3.62 (m, 4H), 1.98-2.06 (m, 3H).

Mass/charge (ESI, +ve) 503.1 (M+H)+.

Table 10

Compounds 10-2 to 10-13 were prepared according to the procedure described in method 10, steps 1-9 above as follows

No. note Chemical structure Name Changes in method Reagent 10-2 γ A 'N' CL .D them: I X, o. ' ° and 1-(4-(7-chloro-6- (2-fluoro-6hydroxyphenyl)- 4-(2methoxyphenyl)phtha lasin-1yl)piperazin-1yl)prop-2-en-1- He - Step 9: 2methoxybenzene boronic acid

- 103 044387

10-3 CK I- I, oh ¹ K ^x VV J. I JLn II Ί G ' _F y. SG'' - ' 1-(4-(7-chloro-4(5-chloro-2methylphenyl)-6(2-fluoro-6hydroxyphenyl)1-phthalazinyl)-1 piperazinyl)-2propen-1-one - Step 9: (5-chloro2methylphenyl)boronic acid (Combi-blocks Inc., San Diego, CA, USA) 10-4 0 II •Г N 1 I 'N ' A / N II 1 11 ¹ 1 II 1-(4-(7-chloro-6(2-fluoro-6hydroxyphenyl)4-(2-(2propanyl)phenyl)1-phthalazinyl)-1 piperazinyl)-2propen-1-one - Step 9: 2isopropylphenylboronic acid (Alfa Aesar, Haver Hill, MA, USA) 10-5 0 oh'/KkAgm ^(L X 1-(4-(7-chloro-4(2-ethylphenyl)-6(2-fluoro-6hydroxyphenyl)1-phthalazinyl)-1 piperazinyl)-2propen-1-one - Step 9: 2ethylbenzeneboronic acid (Alfa Aesar, Haver Hill, MA, USA) 10-6 0 1 ί f N о τ' к и 1 к И _N Г 1 ₍ ρ κ, 1-(4-(7-chloro-6- (2-fluoro-6hydroxyphenyl)4-(4-methyl-3pyridinyl)-1phthalazinyl)-1piperazinyl)-2propen-1-one - Step 9: 4methylpyridine-3boronic acid pinacol ester (carried out at 60°C)

- 104044387

10-7 Ci 1 Λ Ν Μ 1 ^οΗ 'ϊ ίι ^Ν 1 IJ ¹ Ν Ρ . 1' ^[ J 1-(4-(7-chloro-4- (2,6dimethylphenyl)-6(2-φτορ-6hydroxyphenyl)1-phthalazinyl)-1 piperazinyl)-2propen-1-one - Step 9: 2,6dimethylphenylboronic acid (carried out at 80°C) 10-8 ο. 1 τ ,Ν g: 'Ν oS'aaU λ .-L LL .ν 1 ! 1 ' ' ' Ίί '' 1-(4-(7-chloro-6- (2-fluoro-6hydroxyphenyl)4-(2-methyl-3pyridinyl)-1phthalazinyl)-1piperazinyl)-2propen-1-one - Step 9: 2methylpyridine-3boronic acid pinacol ester (Frontier Scientific, Inc., Logan, UT, USA) 10-9 % Μ and Αά fc Α.. ι X ¹ Η 1-(4-(7-chloro-6(2-fluoro-6hydroxyphenyl)4-(1H-indol-4yl)-1phthalazinyl)-1 piperazinyl)-2propen-1-one - Stage 9: 1H- INDOL-4-IL-4boronic acid 10-10 0 IJ ί and οίτ* γ ' J Ν 1 ί J Ν ίι ; ι „ ' ' ΪΙ 1-(4-(7-chloro-4- (2cyclopropylphenyl)-6-(2-fluoro-6hydroxyphenyl)1-phthalazinyl)-1 piperazinyl)-2propen-1-one - Step 9: 2cyclopropylbenzene oronic acid (Combi-Phos Catalysts, Inc., Trenton, NJ, USA)

- 105 044387

10-11 с· Т 'М'^ оР ¹ 4 -/ν 1 1 । and 1 J ) with 1 -(4-(7-chloro-4(2-chlorophenyl)-6(2-fluoro-6hydroxyphenyl)1-phthalazinyl)-1 piperazinyl)-2propen-1-one - Step 9: 2chlorophenylboronic acid (Matrix Scientific, Columbia, SC, USA) 10-12 With ! T Ν [ ''Ν' '1 - 1 OH t* g, Ν i I .N G! I 'Hi ν' 1 -(4-(7-chloro-6(2-fluoro-6hydroxyphenyl)4-(5-methyl-1Nindazol-4-yl)-1phthalazinyl)-!piperazinyl)-2propen-1-one - Step 9: (5-methyl1H-indazol-4yl)boronic acid (Combi-Blocks, Inc., San Diego, CA, USA) 10-13 A—O o ^“7 Uy 4/ and- 1 -(4-(4,7-dichloro6-(2-fluoro-6hydroxyphenyl)- 1-phthal azinyl)-1 piperazinyl)-2propen-1-one excludes Xia stage 9 -

Method 11.

Example 11-1. 6-Chloro-7-(5-methyl-1H-indazol-4-yl)-1-(2-(2-propanyl)phenyl)-4-(4-(2-propenoyl)-1piperazinyl)-2( 1 H)-quinazolinone

Step 1. 4-(1H-Benzo[b] [1,2,3]triazol-1-yl)-7-bromo-6-chloro-1 -(2-isopropylphenyl)quinazolin-2(1H)-one .

Phosphorus oxychloride (1.204 mL, 7.85 mmol) was added to a stirred mixture of 7-bromo-6-chloro-1(2-isopropylphenyl)quinazoline-2,4(1H,3H)-dione (intermediate F, 515 mg, 1.308 mmol), triethylamine (3.31 ml, 23.55 mmol) and 1H-benzo[d][1,2,3]triazole (2.01 g, 16.87 mmol) in acetonitrile (15 ml). The reaction mixture was heated to 80°C and stirred for 1 hour. The reaction mixture was cooled to room temperature. and filtered. The filtrate was then slowly poured into rapidly stirred water (150 ml) at ~10°C. The aqueous suspension was stirred for 15 min before extracting twice with EtOAc (150 ml). The organic layers were combined, washed with brine (150 ml), dried over MgSO ₄ , filtered and concentrated in vacuo to give crude 4-(1H-benzo[d][1,2,3]triazol-1-yl)- 7-bromo-6-chloro-1-(2-isopropylphenyl)quinazolin-2(1H)-one.

Mass/charge (ESI) M+H: 494.0.

- 106 044387

Step 2. tert-Butyl-4-(7-bromo-6-chloro-1-(2-isopropylphenyl)-2-oxo-1,2-dihydroquinazolin-4yl)piperazine-1-carboxylate.

tert-Butylpiperazine-1-carboxylate (268 mg, 1.438 mmol) was added to a stirred mixture of crude 4-(1H-benzo[d][1,2,3]triazol-1-yl)-7-bromo-6-chloro -1-(2-isopropylphenyl)quinazolin-2(1H)one (647 mg, 1.308 mmol) and triethylamine (3.68 ml, 26.2 mmol) in dimethyl sulfoxide (6 ml). The reaction mixture was stirred at 80°C for 30 minutes. The reaction mixture was diluted with EtOAc (100 ml) and washed with water (75 ml). The organic layer was separated, washed with brine (75 ml), dried over MgSO ₄ , filtered and concentrated in vacuo. Chromatographic purification of the residue (silica gel, 0 to 100% EtOAc in heptane) provided tert-butyl-4-(7-bromo-6-chloro-1-(2-isopropylphenyl)-2-oxo-1,2-dihydroquinazoline- 4-yl)piperazine-1carboxylate.

^1H NMR (400 MHz, CHLOROFORM-d) δ: 7.79 (1H, s), 7.49-7.59 (2H, m), 7.36-7.42 (1H, m), 7, 11 (1H, d, J=7.63 Hz) 6.80 (1H, s) 3.79-3.92 (4H, m) 3.62-3.73 (4H, m) 2.60 (1H , spt, J=6.80 Hz) 1.49-1.54 (9H, m) 1.22 (3H, d, J=6.85 Hz) 1.08 (3H, d, J=6.85 Hz), mass/charge (ESI) M+H: 561.0.

Step 3. tert-Butyl-4-(6-chloro-1-(2-isopropylphenyl)-7-(5-methyl-1H-indazol-4-yl)-2-oxo-1,2dihydroquinazolin-4-yl) piperazine-1-carboxylate.

tert-Butyl-4-(7-bromo-6-chloro-1-(2-isopropylphenyl)-2-oxo-1,2-dihydroquinazolin-4-yl)piperazine 1-carboxylate (115 mg, 0.205 mmol), 4- borono-5-methyl-1H-indazole (0.144 ml, 0.819 mmol, Ark Pharm Inc., Arlington Heights, IL, USA), Sphos Pd G3 (0.016 ml, 0.020 mmol) and sodium carbonate (2 M aqueous solution, 0.409 ml, 0.819 mmol) were mixed in 1,2-dimethoxyethane (1 ml) under argon in a sealed vial. The reaction mixture was stirred at 100°C for 24 hours. The reaction mixture was cooled to room temperature. and diluted with EtOAc (50 ml) and water (40 ml). The organic layer was separated, washed with brine (40 ml), dried over MgSO4, filtered and concentrated in vacuo. Chromatographic purification of the residue (silica gel, 0 to 50% (3:1 EtOAc/EUN) in heptane) provided tert-butyl-4-(6-chloro-1-(2-isopropylphenyl)-7-(5methyl-1H- indazol-4-yl)-2-oxo-1,2-dihydroquinazolin-4-yl)piperazine-1-carboxylate.

Mass/charge (ESI) M+H: 613.2.

Step 4. 6-Chloro-1-(2-isopropylphenyl)-7-(5-methyl-1H-indazol-4-yl)-4-(piperazin-1yl)quinazolin-2(1H)-one.

Trifluoroacetic acid (0.5 mL, 6.71 mmol) was added to the stirred mixture of tert-butyl-4(6-chloro-1-(2-isopropylphenyl)-7-(5-methyl-1H-indazol-4-yl) -2-oxo-1,2-dihydroquinazolin-4yl)piperazine-1-carboxylate (78 mg, 0.127 mmol) in dichloromethane (1 ml). The reaction mixture was stirred at room temperature. for 1 hour. The reaction mixture was concentrated in vacuo to give crude 6-chloro-1-(2-isopropylphenyl)-7-(5-methyl-1H-indazol-4-yl)-4-(piperazin-1-yl) quinazolin2(1H)-one.

Mass/charge (ESI) M+H: 513.2.

Step 5. 6-Chloro-7-(5-methyl-1H-indazol-4-yl)-1-(2-(2-propanyl)phenyl)-4-(4-(2-propenoyl)-1piperazinyl)- 2(1H)-quinazolinone.

Acryloyl chloride (10.33 μL, 0.127 mmol) was added to the stirred mixture of 6-chloro-1-(2-isopropylphenyl)-7-(5-methyl-1H-indazol-4-yl)-4-(piperazin-1-yl )quinazolin-2(1H)-one (65 mg, 0.127 mmol) and triethylamine (0.178 ml, 1.267 mmol) in dichloromethane (2 ml) at 0°C. The reaction mixture was stirred at 0°C for 20 minutes. Additional acryloyl chloride (5.17 μL, 0.064 mmol) was added and the reaction mixture was stirred at 0° C. for an additional 20 minutes. The reaction mixture was diluted with DCM (25 ml) and quenched with saturated aqueous sodium bicarbonate (20 ml). The organic layer was separated, dried over MgSO4, filtered and concentrated in vacuo. Chromatographic purification of the residue (silica gel, 0 to 80% (3:1 EtOAc/EtOH) in heptane) provided the crude product. Additional chromatographic purification of the crude product (silica gel, 0 to 100% acetone in heptane) provided separated diastereomers. 6-Chloro-7-(5-methyl-1H-indazol-4-yl)-1-(2-(2-propanyl)phenyl)4-(4-(2-propenoyl)-1-piperazinyl)-2( 1H)-quinazolinone (Example 11-1-1) was the first diastereomer to elute.

1H NMR (400 MHz, CHLOROFORM-d) δ: 10.28 (1H, br s), 7.94 (1H, s), 7.35-7.49 (4H, m), 7.25-7, 31 (2H, m), 7.11 (1H, d, J=7.67 Hz), 6.64 (1H, dd, J=16.79, 10.57 Hz), 6.54 (1H, s ), 6.41 (1H, dd, J=16.79, 1.87 Hz), 5.81 (1H, dd, J=10.57, 1.66 Hz), 3.83-4.07 ( 8H, m), 2.74 (1H, spt, J=6.84 Hz), 2.13 (3H, s), 1.23 (3H, d, J=6.84 Hz), 1.04 ( 3H, d, J=6.84 Hz).

Mass/charge (ESI) M+H: 567.2.

The second eluting diastereomer was further purified by column chromatography (silica gel, 0 to 80% (3:1 EtOAc/EtOH) in heptane) to give 6-chloro-7-(5-methyl-1H-indazol4-yl)-1 -( 2-(2-propanyl)phenyl)-4-(4-(2-propenoyl)-1-piperazinyl)-2(1H)-quinazolinone (Example 11-1-2).

1H NMR (400 MHz, CHLOROFORM-d) δ: 10.37 (1H, br s), 7.94 (1H, s), 7.34-7.50 (4H, m), 7.21-7, 31 (2H, m), 7.13 (1H, d, J=7.67 Hz), 6.64 (1H, dd, J=16.90, 10.68 Hz), 6.55 (1H, s ), 6.41 (1H, dd, J=16.79, 1.66 Hz),

- 107 044387

5.81 (1H, dd, J=10.47, 1.55 Hz), 3.83-4.08 (8H, m), 2.70 (1H, spt, J=6.84 Hz), 2 .13 (3H, s), 1.22 (3H, d,

J=6.84 Hz), 1.03 (3H, d, J=6.84 Hz).

Mass/charge (ESI) M+H: 567.2.

Method 11.

Stages 1 -5 above c. according to the following.

No. Chemical Name Reagent Isomer approx. structure Stage 2: (S)-4- 6-chloro-7-(5- n-boc-2- methyl-1H- methylpiperazine O indazol-4-yl)-4- (CNH Ao A ((2 8)-2-methyl-4- Technologies, f Ί 1st (2-propenoyl)-1 - Inc., Woburn, 11-2-1 elutable 1 JI A 1 NH piperazinyl)-1 - Massachusetts, ^Ax: (2-(2- USA), isomeric mixture propanyl)phenyl)- carried out 2(1H)- at room temperature quinazolinone pace, during nights. Stage 2: (S)-4- 6-chloro-7-(5- n-boc-2- methyl-1H- methylpiperazine 11 A indazol-4-yl)-4- (CNH A ((2 8)-2-methyl-4- Technologies, f Ί 2nd A.C. (2-propenoyl)-1 - Inc., Woburn, 11-2-2 _Ν %γι _N elutable 1 AA A nh piperazinyl)-1 - Massachusetts, 1 1 JO isomeric mixture Ί ι (2-(2- USA), propanyl)phenyl)- carried out 2(1H)- at room temperature quinazolinone pace, during nights.

Selected examples.

Example 12.

1-(4-(7-Chloro-4-cyclopropyl-6-(2-fluoro-6-hydroxyphenyl)-1-phthalazinyl)-1-piperazinyl)-2-propen1-one

Step 1. tert-Butyl-4-(6-(2-((tert-butyldiphenylsilyl)oxy)-6-fluorophenyl)-7-chloro-4-cyclopropylphthalazin-1-yl)piperazine-1-carboxylate.

Into a 20 ml vial filled with tert-butyl 4-(6-(2-((tert-butyldiphenylsilyl)oxy)-6-fluorophenyl)4,7-dichlorophthalazin-1-yl)piperazine-1-carboxylate (intermediate H, 0.060 g, 0.082 mmol), [1,1'-bis-(diphenylphosphino)ferrocene]dichloropalladium(II) complexed with dichloromethane (0.033 g, 0.041 mmol) and 2-methyltetrahydrofuran (2.0 ml) was added. The resulting mixture was sealed and stirred at room temperature. for 10 min before adding cyclopropylzinc bromide (0.5 M in THF, 0.820 mL, 0.410 mmol; Rieke Metals, Lincoln, NE, USA) using a syringe. Reactionary

- 108 044387 the mixture was heated at 80°C for 3 hours before cooling to room temperature. and partitioned between EtOAc (30 ml) and water (10 ml). The aqueous layer was extracted again with EtOAc (20 ml). The combined organic layers were dried over MgSO ₄ , filtered and concentrated in vacuo. The crude product was purified by column chromatography (24 g silica gel, 0 to 30% acetone in heptane) to give tert-butyl-4-(6-(2-((tert-butyldiphenylsilyl)oxy)-6-fluorophenyl)-7- chloro-4-cyclopropylphthalazin-1-yl)piperazine-1-carboxylate.

^1H NMR (CHLOROFORM-d) δ: 8.31-8.38 (m, 1H), 8.15-8.23 (m, 1H), 7.55-7.64 (m, 4H), 7 .39-7.47 (m, 2H), 7.29-7.38 (m, 4H), 6.99-7.09 (m, 1H), 6.74-6.85 (m, 1H) , 6.36-6.47 (m, 1H), 3.68-3.79 (m, 4H), 3.37-3.51 (m, 4H), 2.37-2.48 (m, 1H), 1.48-1.54 (m, 9H), 1.37-1.45 (m, 1H), 1.30-1.33 (m, 1H), 1.00-1.15 ( m, 2H), 0.610.71 (m, 9H).

Mass/charge (ESI) M+H: 737.4.

Step 2. 6-(2-((tert-Butyldiphenylsilyl)oxy)-6-fluorophenyl)-7-chloro-4-cyclopropyl-1-(piperazin1-yl)phthalazine.

Trifluoroacetic acid (0.316 ml, 4.10 mmol) was added to the solution of tert-butyl-4-(6-(2-((tert-butyldiphenylsilyl)oxy)-6-fluorophenyl)-7-chloro-4-cyclopropylphthalazin-1-yl) piperazine-1carboxylate in DCM (0.7 ml). The resulting mixture was sealed and stirred at room temperature. within 30 min. The reaction mixture was diluted with DCM (10 ml) and increased in basicity using saturated aqueous NaHCO ₃ solution (5 ml). The aqueous layer was extracted again with DCM (10 ml). The combined organic layers were dried over MgSO ₄ , filtered and concentrated in vacuo to give 6-(2-((tert-butyldiphenylsilyl)oxy)-6-fluorophenyl)-7-chloro-4-cyclopropyl-1(piperazin-1-yl) phthalazine.

G NMR (CHLOROFORM-d) δ: 8.30-8.36 (m, 1H), 8.18-8.24 (m, 1H), 7.55-7.64 (m, 4H), 7. 40-7.46 (m, 2H), 7.33 (q, J=7.1 Hz, 4H), 6.97-7.09 (m, 1H), 6.74-6.83 (m, 1H), 6.36-6.46 (m, 1H), 3.45-3.55 (m, 4H), 3.163.26 (m, 4H), 2.35-2.49 (m, 1H) , 1.37-1.46 (m, 1H), 1.30-1.33 (m, 1H), 1.06-1.12 (m, 2H), 0.61-0.70 (m, 9H).

Mass/charge (ESI) M+H: 637.2.

Step 3. 1-(4-(6-(2-((tert-Butyldiphenylsilyl)oxy)-6-fluorophenyl)-7-chloro-4-cyclopropylphthalazin-1-yl)piperazin-1-yl)prop-2- en-1-on.

In a 20 ml vial filled with 6-(2-((tert-butyldiphenylsilyl)oxy)-6-fluorophenyl)-7-chloro-4cyclopropyl-1-(piperazin-1-yl)phthalazine (0.023 g, 0.036 mmol), triethylamine (16 μl, 0.114 mmol) and dichloromethane (1.0 ml) were added. The resulting mixture was sealed and stirred at room temperature. for 10 min before adding acryloyl chloride (4.0 μL, 0.049 mmol) using a syringe. The reaction mixture was sealed and continued stirring at room temperature. within 20 min. The reaction mixture was quenched with saturated aqueous NaHCO ₃ (3 ml) and diluted with DCM (10 ml). The aqueous layer was extracted again with DCM (5 ml). The combined organic layers were dried over MgSO4, filtered and concentrated in vacuo to give 1-(4-(6-(2-((tert-butyldiphenylsilyl)oxy)-6-fluorophenyl)-7-chloro-4-cyclopropylphthalazin-1-yl)piperazine -1-yl)prop-2en-1-one.

G NMR (CHLOROFORM-d) δ: 8.32-8.38 (m, 1H), 8.16-8.24 (m, 1H), 7.55-7.65 (m, 4H), 7. 40-7.48 (m, 2H), 7.31-7.38 (m, 4H), 6.98-7.10 (m, 1H), 6.75-6.84 (m, 1H), 6.60-6.72 (m, 1H), 6.41-6.47 (m, 1H), 6.31-6.40 (m, 1H), 5.72-5.82 (m, 1H ), 3.79-4.08 (m, 4H), 3.44-3.62 (m, 4H), 2.38-2.49 (m, 1H), 1.40-1.45 (m , 1H), 1.331.37 (m, 1H), 1.04-1.13 (m, 2H), 0.62-0.68 (m, 9H).

Mass/charge (ESI) M+H: 691.2.

Step 4. 1-(4-(7-Chloro-4-cyclopropyl-6-(2-fluoro-6-hydroxyphenyl)-1-phthalazinyl)-1-piperazinyl)2-propen-1-one.

In a 20 ml bottle filled with 1-(4-(6-(2-((tert-butyldiphenylsilyl)oxy)-6-fluorophenyl)-7-chloro-4cyclopropylphthalazin-1-yl)piperazin-1-yl)prop- 2-en-1-one (0.022 g, 0.032 mmol), tetrahydrofuran (2.0 ml) was added followed by the addition of tetrabutylammonium fluoride solution (1.0 M solution in THF, 0.070 ml, 0.070 mmol). The bottle was sealed and stirred at room temperature. within 30 min. The reaction mixture was concentrated under vacuum. The crude product was purified by column chromatography (24 g silica, 0 to 5% MeOH in DCM) to give 1-(4-(7-chloro-4cyclopropyl-6-(2-fluoro-6-hydroxyphenyl)-1-phthalazinyl )-1-piperazinyl)-2-propen-1-one.

^1H NMR (CHLOROFORM-d) δ: 8.30-8.37 (m, 1H), 8.11-8.18 (m, 1H), 7.29-7.38 (m, 1H), 6 .96-7.18 (m, 1H), 6.88-6.94 (m, 1H), 6.76-6.85 (m, 1H), 6.59-6.72 (m, 1H) , 6.31-6.42 (m, 1H), 5.73-5.84 (m, 1H), 3.73-4.05 (m, 4H), 3.35-3.62 (m, 4H), 2.40-2.52 (m, 1H), 1.35-1.42 (m, 1H), 1.29-1.34 (m, 1H), 1.03-1.14 ( m, 2H).

Mass/charge (ESI) M+H: 453.2.

- 109 044387

Example 13.

1-(4-(4-Anilino-7-chloro-6-(2-fluoro-6-hydroxyphenyl)-1-phthalazinyl)-1-piperazinyl)-2-propen-1-one

Step 1. tert-Butyl-4-(6-(2-((tert-butyldiphenylsilyl)oxy)-6-fluorophenyl)-7-chloro-4-(phenylamino)phthalazin-1 -yl)piperazine-1-carboxylate.

In a 20 ml vial filled with tert-butyl 4-(6-(2-((tert-butyldiphenylsilyl)oxy)-6-fluorophenyl)4,7-dichlorophthalazin-1-yl)piperazine-1-carboxylate (0.060 g , 0.082 mmol), dimethyl sulfoxide (2.0 ml) was added followed by the addition of aniline (0.075 ml, 0.820 mmol). The vial was sealed and refluxed at 80°C for 3 hours. The reaction mixture was cooled to room temperature. and partitioned between EtOAc (30 ml) and water (10 ml). The organic layer was separated and washed with water (2x10 ml). The organic layer was dried over MgSO ₄ , filtered and concentrated in vacuo. The crude product was purified by column chromatography (40 g silica, 0 to 30% EtOAc in heptane) to give tert-butyl-4-(6-(2-((tert-butyldiphenylsilyl)oxy)-6fluorophenyl)-7-chloro -4-(phenylamino)phthalazin-1-yl)piperazine-1-carboxylate.

1H NMR (CHLOROFORM-d) δ: 8.17-8.25 (m, 1H), 7.76-7.81 (m, 1H), 7.60-7.69 (m, 5H), 7. 50-7.55 (m, 2H), 7.40-7.46 (m, 2H), 7.31-7.37 (m, 5H), 7.06-7.11 (m, 2H), 6.76-6.83 (m, 1H), 6.57-6.66 (m, 1H), 6.39-6.50 (m, 1H), 3.66-3.81 (m, 4H ), 3.32-3.43 (m, 4H), 1.51-1.53 (m, 9H), 0.69-0.75 (m, 9H).

Mass/charge (ESI) M+H: 788.2.

Step 2. 7-(2-((tert-Butyldiphenylsilyl)oxy)-6-fluorophenyl)-6-chloro-N-phenyl-4-(piperazin-1yl)phthalazine-1-amine.

Analogously to example 12, stage 2, reaction of tert-butyl-4-(6-(2-((tert-butyldiphenylsilyl)oxy)-6fluorophenyl)-7-chloro-4-(phenylamino)phthalazin-1-yl)piperazine-1 -carboxylate provided the production of 7(2-((tert-butyldiphenylsilyl)oxy)-6-fluorophenyl)-6-chloro-K-phenyl-4-(piperazin-1 -yl)phthalazine-1-amine.

1H NMR (CHLOROFORM-d) δ: 8.19-8.26 (m, 1H), 7.75-7.80 (m, 1H), 7.60-7.68 (m, 5H), 7. 49-7.55 (m, 2H), 7.39-7.46 (m, 3H), 7.32-7.37 (m, 5H), 7.02-7.11 (m, 2H), 6.75-6.84 (m, 1H), 6.59-6.67 (m, 1H), 6.43-6.53 (m, 1H), 3.35-3.47 (m, 4H ), 3.16-3.27 (m, 4H), 0.70-0.76 (m, 9H).

Mass/charge (ESI) M+H: 688.2.

Step 3. 1-(4-(6-(2-((tert-Butyldiphenylsilyl)oxy)-6-fluorophenyl)-7-chloro-4-(phenylamino)phthalazin-1-yl)piperazin-1-yl)prop -2-en-1-on.

Analogously to Example 12, Step 3, the reaction of 7-(2-((tert-butyldiphenylsilyl)oxy)-6-fluorophenyl)-6chloro-N-phenyl-4-(piperazin-1-yl)phthalazin-1-amine provided 1 -(4-(6-(2-((tert-butyldiphenylsilyl)oxy)-6-fluorophenyl)-7-chloro-4-(phenylamino)phthalazin-1-yl)piperazin-1-yl)prop-2-ene -1-she.

1H NMR (CHLOROFORM-d) δ: 8.16-8.24 (m, 1H), 7.77-7.84 (m, 1H), 7.62-7.67 (m, 4H), 7. 52-7.55 (m, 1H), 7.41-7.46 (m, 3H), 7.32-7.38 (m, 6H), 7.02-7.11 (m, 2H), 6.77-6.84 (m, 1H), 6.65-6.71 (m, 1H), 6.46-6.51 (m, 1H), 6.30-6.39 (m, 2H ), 5.73-5.81 (m, 1H), 3.86-4.05 (m, 4H), 3.37-3.53 (m, 4H), 0.69-0.75 (m , 9H).

Mass/charge (ESI) M+H: 742.3.

Step 4. 1-(4-(4-Anilino-7-chloro-6-(2-fluoro-6-hydroxyphenyl)-1-phthalazinyl)-1-piperazinyl)-2propen-1-one.

Analogously to example 12, step 4, reaction 1-(4-(6-(2-((tert-butyldiphenylsilyl)oxy)-6fluorophenyl)-7-chloro-4-(phenylamino)phthalazin-1-yl)piperazin-1- yl)prop-2-en-1-one provided the production of 1-(4-(4-anilino-7-chloro-6-(2-fluoro-6-hydroxyphenyl)-1-phthalazinyl)-1-piperazinyl)-2 -propen-1one.

1H NMR (CHLOROFORM-d) δ: 7.96-8.09 (m, 2H), 7.46-7.57 (m, 2H), 7.37-7.44 (m, 1H), 7. 29-7.33 (m, 1H), 7.20-7.26 (m, 1H), 6.96-7.07 (m, 1H), 6.81-6.87 (m, 1H), 6.70-6.77 (m, 1H), 6.54-6.67 (m, 1H), 6.29-6.41 (m, 1H), 5.68-5.82 (m, 1H ), 3.74-3.96 (m, 4H), 3.12-3.43 (m, 4H).

Mass/charge (ESI) M+H: 504.2.

- 110 044387

Example 14.

-(4-(7-Chloro-4-cyclopentyl-6-(2-fluoro-6-hydroxyphenyl)-1-phthalazinyl)-1-piperazinyl)-2-propen1-one

Step 1. tert-Butyl-4-(6-(2-((tert-butyldiphenylsilyl)oxy)-6-fluorophenyl)-7-chloro-4-cyclopentylphthalazin-1 -yl)piperazine-1-carboxylate.

Analogous to Example 12, Step 1, reaction of tert-butyl 4-(6-(2-((tert-butyldiphenylsilyl)oxy)-6fluorophenyl)-4,7-dichlorophthalazin-1-yl)piperazine-1-carboxylate (intermediate H) and cyclopentylzinc bromide (0.5 M in THF, Rieke Metals, Lincoln, NE) provided tert-butyl-4-(6-(2-((tert-butyldiphenylsilyl)oxy)-6-fluorophenyl)-7-chloro- 4-cyclopentylphthalazin-1yl)piperazine-1-carboxylate.

1H NMR (CHLOROFORM-d) δ: 8.18-8.22 (m, 1H), 8.12-8.16 (m, 1H), 7.60-7.66 (m, 2H), 7. 50-7.56 (m, 2H), 7.39-7.47 (m, 2H), 7.34-7.38 (m, 2H), 7.28-7.33 (m, 2H), 7.09 (br d, J=1.2 Hz, 1H), 6.75-6.82 (m, 1H), 6.37-6.44 (m, 1H), 3.72-3.78 (m, 4H), 3.44-3.51 (m, 4H), 2.03-2.23 (m, 4H), 1.87-1.96 (m, 2H), 1.67-1 .79 (m, 3H), 1.51-1.54 (m, 9H), 0.62-0.67 (m, 9H).

Mass/charge (ESI) M+H: 765.2.

Step 2. 6-(2-((tert-Butyldiphenylsilyl)oxy)-6-fluorophenyl)-7-chloro-4-cyclopentyl-1-(piperazin1-yl)phthalazine.

Similar to Example 12, Step 2, the reaction of tert-butyl-4-(6-(2-((tert-butyldiphenylsilyl)oxy)-6fluorophenyl)-7-chloro-4-cyclopentylphthalazin-1-yl)piperazine-1-carboxylate provided preparation of 6-(2-((tert-butyldiphenylsilyl)oxy)-6-fluorophenyl)-7-chloro-4-cyclopentyl-1-(piperazin-1yl)phthalazine.

1H NMR (CHLOROFORM-d) δ: 8.17-8.21 (m, 1H), 8.12-8.16 (m, 1H), 7.61-7.66 (m, 2H), 7. 51-7.56 (m, 2H), 7.40-7.46 (m, 2H), 7.34-7.38 (m, 2H), 7.29-7.33 (m, 2H), 6.99-7.08 (m, 1H), 6.74-6.82 (m, 1H), 6.37-6.45 (m, 1H), 3.58-3.67 (m, 4H ), 3.27-3.36 (m, 4H), 2.18-2.22 (m, 1H), 2.08-2.12 (m, 2H), 1.86-1.91 (m , 3H), 1.691.77 (m, 3H), 0.59-0.67 (m, 9H).

Mass/charge (ESI) M+H: 665.2.

Step 3. 1-(4-(6-(2-((tert-Butyldiphenylsilyl)oxy)-6-fluorophenyl)-7-chloro-4-cyclopentylphthalazin-1-yl)piperazin-1-yl)prop-2- en-1-on.

Analogous to Example 12, Step 3, the reaction of 6-(2-((tert-butyldiphenylsilyl)oxy)-6-fluorophenyl)-7chloro-4-cyclopentyl-1-(piperazin-1-yl)phthalazine provided 1-(4- (6-(2-((tert-butyldiphenylsilyl)oxy)-6-fluorophenyl)-7-chloro-4-cyclopentylphthalazin-1-yl)piperazin-1-yl)prop-2-en-1one.

1H NMR (CHLOROFORM-d) δ: 8.18-8.25 (m, 1H), 8.13-8.17 (m, 1H), 7.61-7.67 (m, 2H), 7. 50-7.57 (m, 2H), 7.39-7.48 (m, 2H), 7.28-7.37 (m, 4H), 6.99-7.10 (m, 1H), 6.75-6.83 (m, 1H), 6.62-6.71 (m, 1H), 6.33-6.43 (m, 2H), 5.73-5.81 (m, 1H ), 3.84-4.07 (m, 4H), 3.71-3.82 (m, 1H), 3.49-3.65 (m, 4H), 1.80-1.96 (m , 4H), 1.671.77 (m, 4H), 0.62-0.67 (m, 9H).

Mass/charge (ESI) M+H: 719.2.

Step 4. 1-(4-(7-Chloro-4-cyclopentyl-6-(2-fluoro-6-hydroxyphenyl)-1-phthalazinyl)-1-piperazinyl)2-propen-1-one.

Analogous to example 12, step 4, reaction 1-(4-(6-(2-((tert-butyldiphenylsilyl)oxy)-6fluorophenyl)-7-chloro-4-cyclopentylphthalazin-1-yl)piperazin-1-yl)prop -2-en-1-one provided the production of 1-(4-(7-chloro-4-cyclopentyl-6-(2-fluoro-6-hydroxyphenyl)-1-phthalazinyl)-1-piperazinyl)-2-propen- 1she.

1H NMR (CHLOROFORM-d) δ: 8.10-8.22 (m, 2H), 7.29-7.38 (m, 1H), 6.86-6.93 (m, 1H), 6, 77-6.85 (m, 1H), 6.61-6.72 (m, 1H), 6.33-6.44 (m, 1H), 5.74-5.85 (m, 1H), 3.82-4.05 (m, 4H), 3.75-3.82 (m, 1H), 3.40-3.63 (m, 4H), 2.06-2.24 (m, 4H ), 1.81-1.96 (m, 2H), 1.67-1.79 (m, 2H).

- 111 044387

Mass/charge (ESI) M+H: 481.2.

Example 15.

-(4-(7-Chloro-6-(2-fluoro-6-hydroxyphenyl)-4-(1-piperidinyl)-1-phthalazinyl)-1-piperazinyl)-2propen-1-one

Step 1. tert-Butyl-4-(7-chloro-6-(2-fluoro-6-hydroxyphenyl)-4-(piperidin-1-yl)phthalazin-1yl)piperazine-1-carboxylate.

Into a 20 ml vial filled with tert-butyl 4-(6-(2-((tert-butyldiphenylsilyl)oxy)-6-fluorophenyl)4,7-dichlorophthalazin-1-yl)piperazine-1-carboxylate (intermediate H, 0.060 g, 0.082 mmol), piperidine (1.0 mL, 10.10 mmol) was added. The vial was sealed and heated at 80°C for 2 hours. The reaction mixture was cooled to room temperature. and partitioned between EtOAc (30 ml) and water (10 ml). The organic layer was separated and washed with water (2x10 ml). The combined organic layers were dried over MgSO ₄ , filtered and concentrated in vacuo to give tert-butyl-4-(7chloro-6-(2-fluoro-6-hydroxyphenyl)-4-(piperidin-1-yl)phthalazin-1-yl )piperazine-1-carboxylate.

1H NMR (CHLOROFORM-d) δ: 8.09-8.14 (m, 1H), 7.97-8.03 (m, 1H), 7.28-7.35 (m, 1H), 6, 75-6.88 (m, 2H), 3.65-3.76 (m, 4H), 3.30-3.44 (m, 8H), 1.72-1.81 (m, 4H), 1.61-1.71 (m, 3H), 1.48-1.53 (m, 9H).

Mass/charge (ESI) M+H: 542.2.

Step 2. 2-(7-Chloro-1-(piperazin-1-yl)-4-(piperidin-1-yl)phthalazin-6-yl)-3-fluorophenol. Similar to Example 12, Step 2, the reaction of tert-butyl 4-(7-chloro-6-(2-fluoro-6-hydroxyphenyl)-4-(piperidin-1yl)phthalazin-1-yl)piperazine-1-carboxylate provided obtaining 2-(7-chloro-1-(piperazin-1-yl)-4(piperidin-1-yl)phthalazin-6-yl)-3-fluorophenol.

^1H NMR (CHLOROFORM-d) δ: 8.09-8.13 (m, 1H), 7.95-8.03 (m, 1H), 7.28-7.38 (m, 1H), 6 .83-6.89 (m, 1H), 6.75-6.82 (m, 1H), 3.39-3.48 (m, 4H), 3.31-3.38 (m, 4H) , 3.12-3.21 (m, 4H), 1.75-1.80 (m, 4H), 1.64-1.69 (m, 2H).

Mass/charge (ESI) M+H: 442.2.

Step 3. 1-(4-(7-Chloro-6-(2-fluoro-6-hydroxyphenyl)-4-(1-piperidinyl)-1-phthalazinyl)-1piperazinyl)-2-propen-1-one. Analogous to Example 12, Step 3, the reaction of 2-(7-chloro-1-(piperazin-1-yl)-4(piperidin-1-yl)phthalazin-6-yl)-3-fluorophenol provided 1-(4- (7-chloro-6-(2-fluoro-6hydroxyphenyl)-4-(1-piperidinyl)-1-phthalazinyl)-1-piperazinyl)-2-propen-1-one.

1H NMR (CHLOROFORM-d) δ: 8.08-8.15 (m, 1H), 7.98-8.05 (m, 1H), 7.29-7.39 (m, 1H), 6, 86-6.94 (m, 1H), 6.76-6.85 (m, 1H), 6.59-6.70 (m, 1H), 6.30-6.43 (m, 1H), 5.72-5.84 (m, 1H), 3.77-4.05 (m, 4H), 3.40-3.56 (m, 4H), 3.32-3.38 (m, 4H ), 1.73-1.85 (m, 4H), 1.64-1.70 (m, 2H).

Mass/charge (ESI) M+H: 496.2.

- 112 044387

Example 16.

-(4-(7-Chloro-6-(2-fluoro-6-hydroxyphenyl)-4-phenoxy-1-phthalazinyl)-1-piperazinyl)-2-propen-1-one

Step 1. tert-Butyl 4-(7-chloro-6-(2-fluoro-6-hydroxyphenyl)-4-phenoxyphthalazin-1-yl)piperazine-1carboxylate.

Phenol (0.130 g, 1.381 mmol) and tetrahydrofuran (3.0 mL) were charged into a dry 50 mL round bottom flask.

The mixture was cooled to 0°C before potassium tert-butoxide (0.153 g, 1.367 mmol) was added. The mixture was stirred at 0°C for 10 min before heating to room temperature. and stirred for 30 minutes. Add tert-butyl 4-(6-(2-((tert-butyldiphenylsilyl)oxy)-6-fluorophenyl)-4,7-dichlorophthalazin-1yl)piperazine-1-carboxylate (intermediate H, 0.100 g, 0.137 mmol ) and the resulting mixture was heated at 60°C for 2 hours. The reaction mixture was cooled to room temperature. and extinguished with water. The resulting mixture was partitioned between EtOAc (30 ml) and water (15 ml). The aqueous layer was extracted again with EtOAc (20 ml). The combined organic layers were dried over MgSO ₄ , filtered and concentrated in vacuo. The crude product was purified by column chromatography (40 g silica, 10 to 50% acetone) to give tert-butyl-4-(7-chloro-6-(2-fluoro-6hydroxyphenyl)-4-phenoxyphthalazin-1-yl) piperazine-1-carboxylate.

Mass/charge (ESI) M+H: 551.2.

Step 2. 2-(7-Chloro-4-phenoxy-1-(piperazin-1-yl)phthalazin-6-yl)-3-fluorophenol.

Analogous to Example 12, Step 2, the reaction of 4-(7-chloro-6-(2-fluoro-6-hydroxyphenyl)-4phenoxyphthalazin-1-yl)piperazine-1-carboxylate produced 2-(7-chloro-4-phenoxy -1(piperazin-1-yl)phthalazin-6-yl)-3-fluorophenol.

1H NMR (CHLOROFORM-d) δ: 8.37-8.42 (m, 1H), 8.14-8.19 (m, 1H), 7.37-7.45 (m, 2H), 7. 29-7.34 (m, 1H), 7.19-7.25 (m, 2H), 6.89-6.98 (m, 1H), 6.76-6.87 (m, 4H), 3.36-3.45 (m, 4H), 3.13-3.22 (m, 4H).

Mass/charge (ESI) M+H: 451.2.

Step 3. 1-(4-(7-Chloro-6-(2-fluoro-6-hydroxyphenyl)-4-phenoxy-1-phthalazinyl)-1-piperazinyl)-2propen-1-one. Analogous to Example 12, Step 3, the reaction of 2-(7-chloro-4-phenoxy-1-(piperazin-1yl)phthalazin-6-yl)-3-fluorophenol provided 1-(4-(7-chloro-6- (2-fluoro-6-hydroxyphenyl)-4phenoxy-1-phthalazinyl)-1-piperazinyl)-2-propen-1-one.

1H NMR (CHLOROFORM-d) δ: 8.41-8.45 (m, 1H), 8.17-8.20 (m, 1H), 7.40-7.45 (m, 2H), 7, 28-7.37 (m, 2H), 7.20-7.26 (m, 1H), 6.78-6.87 (m, 2H), 6.59-6.70 (m, 1H), 6.31-6.41 (m, 1H), 5.97-6.06 (m, 1H), 5.74-5.81 (m, 1H), 3.76-4.03 (m, 4H ), 3.38-3.53 (m, 4H).

Mass/charge (ESI) M+H: 505.2.

Examples 17-1 and 17-2.

(2E)-1-(4-(5-Chloro-7-fluoro-6-(3-methoxy-1-naphthalenyl)-2,1-benzothiazol-3-yl)-1-piperazinyl)-4(dimethylamino) -2-buten-1-one (Example 17-1) and (2E)-1-(4-(5-chloro-7-fluoro-6-(3-hydroxy-1-naphthalenyl)-2,1benzothiazol-3 -yl)-1-piperazinyl)-4-(dimethylamino)-2-buten-1-one (Example 17-2)

- 113 044387

Step 1. tert-Butyl-4-(5-chloro-6-(3-methoxynaphthalen-1-yl)benzo[c]isothiazol-3-yl)piperazine-1 carboxylate.

Tert-Butyl 4-(6-bromo-5-chloro-7-fluorobenzo[c]isothiazol-3-yl)piperazine-1-carboxylate suspension (intermediate D, 459 mg, 1.02 mmol), (3- methoxynaphthalen-1-yl)boronic acid (823 mg, 4.07 mmol) and cesium carbonate (1.33 g, 4.07 mmol) in a mixture of 1,4-dioxane (8 ml) and water (2 ml) were degassed with using a flow of argon. Tetrakis(triphenylphosphine)palladium (118 mg, 0.10 mmol) was added and the mixture was again degassed with a stream of argon. The reaction mixture was sealed and heated at 100°C for 23 hours. The reaction mixture was cooled to room temperature, diluted with brine (60 ml) and extracted twice with EtOAc. The combined organic layers were dried over anhydrous sodium sulfate and concentrated. The residue was purified by silica gel chromatography (eluent: 0-2% MeOH in DCM) to give tert-butyl-4-(5-chloro-6-(3methoxynaphthalen-1-yl)benzo[c]isothiazol-3-yl)piperazine -1-carboxylate.

Mass/charge (ESI) M+H: 528.0.

Step 2. 5-Chloro-6-(3-methoxynaphthalen-1-yl)-3-(piperazin-1-yl)benzo[c]isothiazole.

A solution of tert-butyl 4-(5-chloro-6-(3-methoxynaphthalen-1-yl)benzo[c]isothiazol-3-yl)piperazine-1 carboxylate (327 mg, 0.56 mmol) in DCM ( 6 ml) trifluoroacetic acid (1.04 ml, 13.9 mmol) was added using a syringe. The resulting yellow solution was stirred at room temperature. for 4 hours and then concentrated. The residue was purified by silica gel chromatography (eluent: 025% MeOH in DCM) to give TFA 5-chloro-6-(3-methoxynaphthalen-1-yl)-3-(piperazin-1yl)benzo[c]isothiazole monosalt.

Mass/charge (ESI) M+H: 428.0.

Step 3. (2E)-1 -(4-(5-Chloro-7-fluoro-6-(3-methoxy-1-naphthalenyl)-2,1-benzothiazol-3-yl)-1 piperazinyl)-4- (dimethylamino)-2-buten-1-one.

To a solution of 5-chloro-6-(3-methoxynaphthalen-1-yl)-3-(piperazin-1-yl)benzo[c]isothiazole (74 mg TFA monosalt, 0.14 mmol) and trans-4-dimethylaminocrotonic acid hydrochloride (38 mg, 0.23 mmol) in DMA (2 ml), thionyl chloride (41 μl, 0.69 mmol) was added using a syringe. The resulting brown solution was stirred at room temperature. for 2.5 hours. The reaction mixture was quenched with water (50 ml) and extracted with 8:1 DCM/MeOH. The organic layer was dried over anhydrous sodium sulfate and concentrated. The residue was purified by silica gel chromatography (eluent: 0-15% MeOH in DCM) to give (2E)-1-(4-(5-chloro-7-fluoro-6-(3-methoxy-1-naphthalenyl)-2 ,1-benzothiazol-3yl)-1-piperazinyl)-4-(dimethylamino)-2-buten-1-one.

¹ H NMR (400 MHz, DMSO-d ₆ ) δ: 8.10 (s, 1H), 7.94 (d, J=8.4 Hz, 1H), 7.46-7.55 (m, 2H ), 7.27-7.35 (m, 2H), 7.19 (d, J=2.5 Hz, 1H), 6.61-6.72 (m, 2H), 3.94 (s, 3H), 3.80-3.93 (m, 4H), 3.62-3.68 (m, 4H), 3.07 (d, J=4.3 Hz, 2H), 2.18 (s , 6H).

Mass/charge (ESI) M+H: 539.2.

Step 4. (2E)-1-(4-(5-Chloro-7-fluoro-6-(3-hydroxy-1-naphthalenyl)-2,1-benzothiazol-3-yl)-1piperazinyl)-4-( dimethylamino)-2-buten-1-one.

To a solution of (2E)-1 -(4-(5-chloro-7-fluoro-6-(3-methoxy-1-naphthalenyl)-2,1-benzothiazol-3-yl)-1 piperazinyl)-4-( boron tribromide (1.0 M in hexanes, 218 µl, 0 .22 mmol) using a syringe. The resulting yellow slurry was stirred at 0°C for 2.75 hours and then quenched with saturated aqueous NaHCO ₃ solution (4 ml). The mixture was extracted twice with 4:1 DCM/MeOH. The combined organic layers were dried over anhydrous sodium sulfate and concentrated. The residue was purified by silica gel chromatography (eluent: 0-18% MeOH in DCM) to give (2E)-1-(4-(5-chloro-7-fluoro-6-(3-hydroxy-1-naphthalenyl)-2 ,1-benzothiazol-3yl)-1-piperazinyl)-4-(dimethylamino)-2-buten-1-one.

- 114 044387

1H NMR (400 MHz, DMSO-d6) δ: 9.97 (s, 1H), 8.10 (s, 1H), 7.80 (d, J=8.4 Hz, 1H), 7.40- 7.46 (m, 1H),

7.19-7.30 (m, 3H), 7.07 (d, J=2.4 Hz, 1H), 6.62-6.71 (m, 2H), 3.80-3.93 ( m, 4H), 3.62-3.69 (m, 4H), 3.07 (d,

J=4.1 Hz, 2H), 2.17 (s, 6H).

Mass/charge (ESI) M+H: 525.0.

Examples from 18-1 to 18-3.

1-(4-(5-Chloro-7-fluoro-6-(3-methoxy-1-naphthalenyl)-2,1-benzothiazol-3-yl)-1-piperazinyl)-2(hydroxymethyl)-2-propene -1-one (Example 18-1), and 2-(bromomethyl)-1-(4-(5-chloro-7-fluoro-6-(3-hydroxy-1naphthalenyl)-2,1-benzothiazol-3- yl)-1-piperazinyl)-2-propen-1-one (Example 18-2), and 1-(4-(5-chloro-7-fluoro6-(3-hydroxy-1-naphthalenyl)-2,1 -benzothiazol-3-yl)-1-piperazinyl)-2-(hydroxymethyl)-2-propen-1-one (example 18-3)

Step 1. 1 -(4-(5-Chloro-7-fluoro-6-(3-methoxy-1-naphthalenyl)-2,1-benzothiazol-3-yl)-1-piperazinyl)-2(hydroxymethyl)- 2-propen-1-one. A solution of 1-(4-(5-chloro-7-fluoro-6-(3-methoxynaphthalen-1-yl)benzo[c]isothiazol-3-yl)piperazin-1-yl)prop-2-ene was loaded into the bottle -1-one (intermediate O, 29 mg, 0.06 mmol) in tert-butanol (0.4 ml) and water (0.4 ml). Phenol (5.7 mg, 0.06 mmol), DABCO (20.3 mg, 0.18 mmol) and formaldehyde (37% aqueous solution, 24 μl, 0.24 mmol) were added sequentially. The resulting solution was sealed and heated at 55°C for 29 hours. The reaction mixture was cooled to room temperature. and partitioned between water (6 ml) and 10:1 DCM/MeOH. The organic layer was separated and the aqueous layer was extracted two more times with 10:1 DCM/MeOH. The combined organic layers were dried over anhydrous sodium sulfate and concentrated. The residue was purified by silica gel chromatography (eluent: 0-3.5% MeOH in DCM) to give 1-(4-(5-chloro-7-fluoro-6-(3-methoxy-1naphthalenyl)-2,1-benzothiazole -3-yl)-1-piperazinyl)-2-(hydroxymethyl)-2-propen-1-one.

1H NMR (400 MHz, DMSO-d ₆ ) δ: 8.12 (s, 1H), 7.94 (d, J=8.2 Hz, 1H), 7.47-7.55 (m, 2H) , 7.25-7.34 (m, 2H), 7.19 (d, J=2.5 Hz, 1H), 5.43 (br. s, 1H), 5.20 (br. s, 1H ), 5.14 (t, J=5.8 Hz, 1H), 4.12 (d, J=5.7 Hz, 2H), 3.94 (s, 3H), 3.78-3.85 (m, 4H), 3.54-3.66 (m, 4H).

Mass/charge (ESI) M+H: 512.0.

Step 2. 2-(Bromomethyl)-1-(4-(5-chloro-7-fluoro-6-(3-hydroxy-1-naphthalenyl)-2,1-benzothiazol-3-yl)-1piperazinyl)-2 -propen-1-one and 1-(4-(5-chloro-7-fluoro-6-(3-hydroxy-1-naphthalenyl)-2,1-benzothiazol-3-yl)-1piperazinyl)-2-( Hydroxymethyl)-2-propen-1-one.

To a solution of 1 -(4-(5-chloro-7-fluoro-6-(3-methoxy-1-naphthalenyl)-2, 1-benzothiazol-3-yl)-1-piperazinyl)2-(hydroxymethyl)-2 -propen-1-one (17.1 mg, 0.033 mmol) in 1,2-dichloroethane (4 ml) at 0°C, a solution of boron tribromide (1.0 M in hexanes, 167 μl, 0.17 mmol) was added dropwise ) using a syringe. The resulting slurry was stirred at 0°C for 40 min before quenching with saturated aqueous NaHCO ₃ solution (5 ml). The mixture was extracted twice with 4:1 DCM/MeOH. The combined organic layers were dried over anhydrous sodium sulfate and concentrated. The residue was purified by silica gel chromatography (eluent: 0-7% MeOH in DCM) to give two products.

Peak of the first eluted compound: 2-(bromomethyl)-1-(4-(5-chloro-7-fluoro-6-(3-hydroxynaphthalen1-yl)benzo[c]isothiazol-3-yl)piperazin-1-yl) prop-2-en-1-one.

1H NMR (400 MHz, DMSO-d6) δ: 9.96 (br. s, 1H), 8.13 (s, 1H), 7.80 (d, J=8.2 Hz, 1H), 7, 40-7.47 (m, 1H), 7.19-7.29 (m, 3H), 7.07 (d, J=2.4 Hz, 1H), 5.78 (s, 1H), 5 .41 (s, 1H), 4.38 (s, 2H), 3.84-3.93 (m, 4H), 3.62-3.72 (m, 4H).

Mass/charge (ESI) M+H: 560.0.

Peak of the second eluted compound: 1-(4-(5-chloro-7-fluoro-6-(3-hydroxynaphthalen-1yl)benzo[c]isothiazol-3-yl)piperazin-1-yl)-2-(hydroxymethyl) prop-2-en-1-one.

1H NMR (400 MHz, DMSO-d ₆ ) δ: 9.98 (br. s, 1H), 8.11 (s, 1H), 7.79 (d, J=8.2 Hz, 1H), 7 .37-7.48 (m, 1H),

- 115 044387

7.17-7.28 (m, 3H), 7.07 (d, J=2.4 Hz, 1H), 5.43 (br. s, 1H), 5.20 (br. s, 1H) , 5.07-5.14 (m, 1H), 4.12 (br. s, 2H),

3.78-3.86 (m, 4H), 3.57-3.66 (m, 4H).

Mass/charge (ESI) M+H: 498.0.

Examples from 19-1 to 19-3.

-(4-(5-Chloro-7-fluoro-6-(5-methoxy-1 -methyl-1 H-indazol-7-yl)-2,1 -benzothiazol-3 -yl)-1 -piperazinyl)2 -propen-1-one (example 19-1), and 1-(4-(5-chloro-7-fluoro-6-(5-hydroxy-1-methyl-1H-indazol-7-yl)-2, 1-benzothiazol-3-yl)-1-piperazinyl)-2-propen-1-one (Example 19-2), and 1-(4-(5-chloro-7-fluoro-6-(5-hydroxy-2methyl -2H-indazol-7 -yl)-2,1-benzothiazol-3 -yl)-1-piperazinyl)-2-propen-1-one (example 19-3)

Step 1. tert-Butyl-4-(5-chloro-7-fluoro-6-(5-methoxy-1H-indazol-7-yl)benzo[c]isothiazol-3yl)piperazine-1-carboxylate.

A suspension of intermediate D (232 mg, 0.51 mmol), 5-methoxy-7-(4,4,5,5-tetramethyl-1,3,2dioxaborolan-2-yl)-III-indazole (53 5 mg, 1.95 mmol, see Synthesis below) and cesium carbonate (636 mg, 1.95 mmol) in a mixture of 1,4-dioxane (8 ml) and water (2 ml) were degassed using a stream of argon. Tetrakis(triphenylphosphine)palladium (59 mg, 0.05 mmol) was added and the mixture was again degassed with a stream of argon. The reaction mixture was sealed and heated at 100°C for 18 hours. The reaction mixture was cooled to room temperature. and it was partitioned between saline (40 ml) and EtOAc. The aqueous layer was then extracted twice with EtOAc and the combined organic layers were dried over anhydrous sodium sulfate and concentrated. The residue was purified by silica gel chromatography (eluent: 0-4.5% DCM/MeOH) to give tert-butyl-4-(5-chloro-7-fluoro-6-(5-methoxy-1Hindazol-7-yl)benzo [c]isothiazol-3-yl)piperazine-1-carboxylate.

LCMS-ESI (POS.) mass/charge: 518.2 (M+H) ⁺ .

^1H NMR (400 MHz, DMSO- _d6 ) δ: 12.90 (br. s, 1H), 8.06 (s, 1H), 8.03 (s, 1H), 7.31 (d, J =1.4 Hz, 1H), 6.99 (d, J=2.2 Hz, 1H), 3.83 (s, 3H), 3.61-3.69 (m, 4H), 3.54 -3.60 (m, 4H), 1.45 (s, 9H).

- 116 044387

5-Methoxy-7-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-III-indazole.

Suspension of 7-bromo-5-methoxy-1H-indazole (1.00 g, 4.40 mmol, Ark Pharm Inc., Arlington Heights, IL, USA), potassium acetate (1.30 g, 13.2 mmol) and bis-(pinacolato)diboron (1.23 g, 4.84 mmol) in 1,4dioxane (18 ml) were degassed with a stream of argon. [1,1'-Bis-(diphenylphosphino)ferrocene]dichloropalladium(II) complexed with dichloromethane (108 mg, 0.13 mmol) was added and degassed again with a stream of argon. The reaction mixture was sealed and heated at 80°C for 2 days. The reaction mixture was cooled to room temperature. and it was partitioned between water (50 ml) and EtOAc. The aqueous layer was extracted twice with EtOAc and the combined organic layers were dried over anhydrous sodium sulfate and concentrated. The residue was purified by silica gel chromatography (eluent: 2-65% EtOAc/heptane) to give 5-methoxy-7-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H -indazole.

LCMS-ESI (POS.) mass/charge: 275.1 (M+H)+.

Step 2. tert-Butyl-4-(5-chloro-7-fluoro-6-(5-methoxy-1-methyl-1H-indazol-7-yl)benzo[c]isothiazol-3yl)piperazine-1-carboxylate and tert-butyl 4-(5-chloro-7-fluoro-6-(5-methoxy-2-methyl-2H-indazol-7yl)benzo[c]isothiazol-3-yl)piperazine-1-carboxylate.

In a solution of tert-butyl-4-(5-chloro-7-fluoro-6-(5-methoxy-1 H-indazol-7-yl)benzo[c]isothiazol-3 yl)piperazine-1-carboxylate (115 MG , 0.22 mmol) sodium hydride (60% dispersion in mineral oil, 44.5 mg, 1.1 mmol) was added to THF (5 ml). After 10 min, iodomethane (69 µl, 1.1 mmol) was added and the reaction mixture was stirred at room temperature. for an additional 15 min before partitioning between saturated aqueous ammonium chloride (10 mL) and DCM. The aqueous layer was extracted twice with DCM and the combined organic layers were dried over anhydrous sodium sulfate and concentrated to give a mixture of tert-butyl-4-(5-chloro-7-fluoro-6-(5-methoxy-1-methyl-1H-indazole -7yl)benzo[c]isothiazol-3-yl)piperazin-1-carboxylate and tert-butyl-4-(5-chloro-7-fluoro-6-(5-methoxy-2methyl-2H-indazol-7-yl )benzo[c]isothiazol-3-yl)piperazine-1-carboxylate. The crude mixture was used in the next step without purification.

LCMS-ESI (POS.) mass/charge: 532.0 (M+H) ⁺ .

Step 3. 5 -Chloro-7-fluoro-6-(5 -methoxy-1 -methyl-1 H-indazol-7-yl)-3 -(piperazin-1 -yl)benzo[c]isothiazole (intermediate J ) and 5-chloro-7-fluoro-6-(5-methoxy-2-methyl-2H-indazol-7-yl)-3(piperazin-1-yl)benzo[c]isothiazole (intermediate K).

To a solution of a crude mixture consisting of a mixture of tert-butyl-4-(5-chloro-7-fluoro-6-(5-methoxy-1methyl-1 H-indazol-7 -yl)benzo[c]isothiazol-3 -yl )piperazine-1-carboxylate and tert-butyl-4-(5-chloro-7 fluoro-6-(5-methoxy-2-methyl-2H-indazol-7-yl)benzo[c]isothiazol-3-yl) piperazine-1-carboxylate (143 mg) in DCM (6 ml) was added with trifluoroacetic acid (484 µl, 6.5 mmol) using a syringe. The resulting solution was stirred at room temperature. for 25 minutes and then concentrated. The residue was purified by chromatography on silica gel (eluent: 0-25% DCM/MeOH).

Peak of the first eluted compound: TFA Monosol 5-chloro-7-fluoro-6-(5-methoxy-1-methyl-1Hindazol-7-yl)-3-(piperazin-1-yl)benzo[c]isothiazole (intermediate J).

LCMS-ESI (POS.) mass/charge: 432.0 (M+H) ⁺ .

¹ H NMR (400 MHz, DMSO-d ₆ ) δ: 8.16 (s, 1H), 8.03 (s, 1H), 7.35 (d, J=2.4 Hz, 1H), 6, 99 (d, J=2.4 Hz, 1H), 3.84 (s, 3H), 3.67-3.76 (m, 4H), 3.56 (s, 3H), 3.36-3 .42 (m, 4H).

Peak of the second eluted compound: TFA Monosol 5-chloro-7-fluoro-6-(5-methoxy-2-methyl-2Hindazol-7-yl)-3-(piperazin-1-yl)benzo[c]isothiazole (intermediate K).

LCMS-ESI (POS.) mass/charge: 432.0 (M+H) ⁺ .

Step 4. 1 -(4-(5 -Chloro-7-fluoro-6-(5 -methoxy-1 -methyl-1 H-indazol-7-yl)-2,1 -benzothiazol-3 -yl)-1 piperazinyl)-2-propen-1-one.

To an ice-cold slurry of TFA monosalt intermediate J (108 mg, 0.20 mmol) in DCM (5 mL) was added DIPEA (104 μL, 0.60 mmol) followed by dropwise addition of acryloyl chloride (24 μL, 0.30 mmol). using a syringe. The resulting solution was stirred at 0°C for 3 hours and then quenched with saturated aqueous NaHCO ₃ solution (15 ml) and extracted twice with DCM. The combined organic layers were dried over anhydrous sodium sulfate and concentrated. The residue was purified by silica gel chromatography (eluent: 0-7% DCM/MeOH) to give 1-(4-(5-chloro-7-fluoro-6-(5-methoxy-1-methyl-1H-indazole-7 -yl)-2,1-benzothiazol3-yl)-1-piperazinyl)-2-propen-1-one.

LCMS-ESI (POS.) mass/charge: 486.0 (M+H) ⁺ .

1H NMR (400 MHz, DMSO-d ₆ ) δ: 8.13 (s, 1H), 8.02 (s, 1H), 7.33 (d, J=2.2 Hz, 1H), 6.99 (d, J=2.4 Hz, 1H), 6.85 (dd, J=16.6, 10.6 Hz, 1H), 6.18 (dd, J=16.7, 2.3 Hz, 1H), 5.76 (dd, J=10.5, 2.3 Hz, 1H), 3.85-3.95 (m, 4H), 3.84 (s, 3H), 3.62-3 .72 (m, 4H), 3.56 (s, 3H).

- 117 044387

Step 5. 1 -(4-(5-Chloro-7-fluoro-6-(5-hydroxy-1 -methyl-1 H-indazol-7 -yl)-2, 1 -benzothiazol-3 -yl)-1 piperazinyl)-2-propen-1-one.

To an ice-cooled solution of 1-(4-(5-chloro-7-fluoro-6-(5-methoxy-1-methyl-1H-indazol-7yl)benzo[c]isothiazol-3-yl)piperazin-1-yl )prop-2-en-1-one (72.5 mg, 0.15 mmol) in 1,2-dichloroethane (5 ml), a solution of boron tribromide (1.0 M in hexanes, 746 μl, 0. 75 mmol) using a syringe. The resulting slurry was stirred at 0°C for 3.75 hours and then quenched with saturated aqueous NaHCO ₃ (5 ml) and extracted twice with 4:1 DCM/MeOH. The combined organic layers were dried over anhydrous sodium sulfate and concentrated. The residue was purified by silica gel chromatography (eluent: 0-6% DCM/MeOH) to give 1-(4-(5-chloro-7-fluoro-6-(5-hydroxy-1-methyl-1H-indazole-7 -yl)benzo[c]isothiazol-3 -yl)piperazin-1 yl)prop-2-en-1-one. LCMS-ESI (POS.) mass/charge: 472.0 (M+H)+.

1H NMR (400 MHz, DMSO-d6) δ: 9.40 (s, 1H), 8.12 (s, 1H), 7.92 (s, 1H), 7.12 (d, J=2.2 Hz, 1H), 6.81-6.91 (m, 2H), 6.18 (dd, J=16.7, 2.5 Hz, 1H), 5.76 (dd, J=10.4, 2.4 Hz, 1H), 3.81-3.94 (m, 4H), 3.62-3.70 (m, 4H), 3.52 (s, 3H).

Synthesis of 1-(4-(5-chloro-7-fluoro-6-(5-hydroxy-2-methyl-2H-indazol-7-yl)-2,1-benzothiazol-3-yl)-1piperazinyl)-2 -propen-1-one was performed using intermediate K from step 3, steps 4 and 5 as above to give 1-(4-(5-chloro-7-fluoro-6-(5-hydroxy-2- methyl-2H-indazol-7yl)-2,1-benzothiazol-3-yl)-1-piperazinyl)-2-propen-1-one.

LCMS-ESI (POS.) mass/charge: 472.0 (M+H)+.

1H NMR (400 MHz, DMSO-d ₆ ) δ: 9.28 (s, 1H), 8.11 (s, 1H), 8.01 (s, 1H), 6.95 (d, J=2, 0 Hz, 1H), 6.77-6.90 (m, 2H), 6.18 (dd, J=16.7, 2.5 Hz, 1H), 5.76 (dd, J=10.4 , 2.2 Hz, 1H), 4.03 (s, 3H), 3.80-3.94 (m, 4H), 3.58-3.66 (m, 4H).

Example 20.

1-(4-(5-Chloro-7-fluoro-6-(3-hydroxy-1-naphthalenyl)-2,1-benzothiazol-3-yl)-1-piperazinyl)-4hydroxy-2-methylidene-1 - butanone

Step 1. 4-((tert-Butyldiphenylsilyl)oxy)-1-(4-(5-chloro-7-fluoro-6-(3-methoxynaphthalen-1yl)benzo[c]isothiazol-3-yl)piperazine-1 -yl)-2-methylenebutan-1-one.

To a solution of 4-((tert-butyldiphenylsilyl)oxy)-2-methylenebutanoic acid (101 mg, 0.29 mmol, prepared according to Pihko, P.M., J. Org. Chem., 2006, 71, 2538-2541 and Greaney , M.F., Org. Lett., 2007, 9, 1931-1934) in DCM (2 ml) was added a 2 M solution of oxalyl chloride (0.21 ml, 0.43 mmol) at 0°C, followed by the addition of a catalytic amount of DMF (5 µl). The reaction mixture was heated to room temperature. and it was stirred for 2 hours. The reaction mixture was concentrated in vacuo, then diluted with DCM (1 ml) and added to the solution of 5-chloro-7-fluoro-6-(3methoxynaphthalen-1-yl)-3-(piperazin- 1-yl)benzo[c]isothiazole (intermediate N, 122 mg, 0.29 mmol), triethylamine (0.20 ml, 1.43 mmol) and DCM (2 ml). The reaction mixture was heated to room temperature. and DMAP (2 mg, 0.016 mmol) was added. The reaction mixture was stirred at room temperature. for 15 hours, then concentrated in vacuo and purified by silica gel column chromatography (eluent: 0-50% EtOAc:heptanes) to give 4-((tert-butyldiphenylsilyl)oxy)-1-(4-(5chloro-7- fluoro-6-(3-methoxynaphthalen-1-yl)benzo[c]isothiazol-3-yl)piperazin-1-yl)-2-methylenebutan-1-one.

1H NMR (400 MHz, DMSO-d ₆ ) δ: 8.06 (s, 1H), 7.94 (d, J=8.0 Hz, 1H), 7.63-7.61 (m, 4H) , 7.52-7.49 (m, 2H), 7.47-7.40 (m, 6H), 7.33-7.28 (m, 2H), 7.20-7.19 (m, 1H), 5.37 (s, 1H), 5.24 (s, 1H), 3.94 (s, 3H), 3.83-3.76 (m, 6H), 3.53 (br s, 2H), 3.31 (s, 4H), 1.01 (s, 9H).

Mass/charge (ESI) M+H: 764.

Step 2. 1 -(4-(5-Chloro-7-fluoro-6-(3-hydroxy-1-naphthalenyl)-2,1-benzothiazol-3-yl)-1-piperazinyl)4-hydroxy-2- Methylidene-1-butanone.

To a solution of 4-((tert-butyldiphenylsilyl)oxy)-1-(4-(5-chloro-7-fluoro-6-(3-methoxynaphthalen-1- 118 044387 yl)benzo[c]isothiazol-3-yl) piperazin-1-yl)-2-methylenebutan-1-one (85 mg, 0.11 mmol) and DCM (2 ml) was added a 2 M solution of BBr ₃ (0.28 ml, 0.56 mmol) in DCM at 0 °C. The reaction mixture was quenched with water, concentrated in vacuo and purified by silica gel column chromatography (eluting with 0-50% heptane/3:1 EtOAc:EtOH) to give 1-(4-(5-chloro-7-fluoro -6-(3hydroxy-1-naphthalenyl)-2,1-benzothiazol-3-yl)-1-piperazinyl)-4-hydroxy-2-methylidene-1-butanone.

1H NMR (400 MHz, DMSO-d6) δ: 9.93 (br s, 1H), 8.11 (s, 1H), 7.80 (d, J=12 Hz, 1H), 7.43 (m 1H), 7.267.20 (m, 3H), 7.07 (s, 1H), 5.32 (s, 1H), 5.16 (s, 1H), 3.83 (br s, 4H), 3 .63 (br s, 4H), 3.53 (t, J=8.0 Hz, 2H), 2.42 (t, J=8.0 Hz, 2H).

¹⁹ F NMR (377 MHz, DMSO-d ₆ ) δ: -123.8 (s, 1F).

Mass/charge (ESI) M+H: 512.

Example 21.

-(4-(5-Chloro-7-fluoro-6-(7-hydroxy-5-quinolinyl)-2,1-benzothiazol-3-yl)-1-piperazinyl)-2-propen-1-one

1-(4-(5-Chloro-7-fluoro-6-(7-hydroxy-5-quinolinyl)-2,1-benzothiazol-3-yl)-1-piperazinyl)-2-propen-1one was prepared from the intermediate Compound D using Method 1 using 7-methoxy-5(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)quinoline (synthesis below) with the following modifications: Step 7 used S -Phos Pd G3, aqueous solution of potassium carbonate and DME; at stage 8-1 TFA/DCM was used; Step 8-2 used DCE as solvent and Step 8-3 used boron tribromide solution (1.0 M in DCE) to give 1-(4-(5-chloro-7-fluoro-6-(7-hydroxy -5-quinolinyl)-2,1benzothiazol-3-yl)-1-piperazinyl)-2-propen-1-one.

1H NMR (400 MHz, CDCl3) δ: 8.81 (dd, J=4.2, 1.3 Hz, 1H), 7.72-7.78 (m, 2H), 7.64 (s, 1H ), 7.28 (d, J=2.2 Hz, 1H), 7.16 (dd, J=8.4, 4.3 Hz, 1H), 6.56-6.66 (m, 1H) , 6.40 (dd, J=16.8, 1.6 Hz, 1H), 5.78-5.87 (m, 1H), 4.01 (br. s., 2H), 3.89 ( br. s., 2H), 3.50-3.60 (m, 4H).

^19F NMR (376 MHz, CDCl ₃ ) δ: -121.33 (s, 1F).

MS (ESI, +ve) mass/charge: 469.1 (M+1) ⁺ .

I ^Br - _ '/ V IVluWi )-0 |- %

J ~ ⁱⁿ in rz —__- Ά θ AV o- dmi wn cj ;

7-Methoxy-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)quinoline.

5-Bromo-7-methoxyquinoline solution (0.407 g, 1.71 mmol, OxChem, Wood Dale, IL, USA), 4,4,4',4',5,5,5',5'-octamethyl-2 ,2'-bi(1,3,2-dioxaborolane) (0.912 g, 3.59 mmol), PdCl ₂ (dppf (0.051 g, 0.070 mmol) and potassium acetate (0.503 g, 5.13 mmol) in DMF ( 9 ml) was stirred at 90°C for 1 hour, then at 100°C for 45 minutes. The reaction mixture was diluted with EtOAc (100 ml) and washed with saturated aqueous sodium bicarbonate (2x75 ml). The organic layer was separated , dried over anhydrous Na ₂ SO ₄ and concentrated in vacuo.The crude product was adsorbed onto silica and purified by column chromatography (silica gel, 0-80% heptane/EtOAc) to give 7-methoxy-5-(4,4,5, 5-tetramethyl-1,3,2-dioxaborolan-2-yl)quinoline.

MS (ESI, +ve) mass/charge: 286.1 (M+1) ⁺ .

Example 22.

1-(5-Chloro-7-fluoro-6-(3-hydroxy-1-naphthalenyl)-2,1-benzothiazol-3-yl)-4-(2-propenoyl)-2piperazinecarboxylic acid

V V

SI ON

To a solution of methyl 4-acryloyl-1-(5-chloro-7-fluoro-6-(3-hydroxynaphthalen-1-yl)benzo[c]isothiazol-3yl)piperazine-2-carboxylate (example 7-3, 0.022 g, 0.042 mmol) in THF/EtOH (1:1; 6 ml) at 0°C, NaOH (5 N aq.; 1.0 ml, 5.0 mmol) was added and the resulting mixture was stirred at 0°C for 5 minutes. The reaction mixture was acidified with 5N. HCl at 0°C, extracted with EtOAc and purified by HPLC to give 1-(5-chloro-7-fluoro-6-(3-hydroxy-1-naphthalenyl)-2,1-benzothiazol-3-119 044387 yl)-4-(2-propenoyl)-2-piperazinecarboxylic acid.

Mass/charge (ESI, +ve) 512.0 (M+H) ⁺ .

1H NMR (400 MHz, DMSO-d ₆ ) δ: 3.14-3.28 (m, 1H) 3.52-3.87 (m, 3H) 4.15-5.03 (m, 2H) 5 .15-5.23 (m, 1H) 5.77-5.83 (m, 1H) 6.13-6.24 (m, 1H) 6.86 (br. s., 1H), 7.06 -7.12 (m, 1H), 7.20-7.30 (m, 3H), 7.38-7.49 (m, 1H), 7.76-7.84 (m, 1H) 8, 07-8.13 (m, 1H) 9.98 (br. s., 1 H) 13.42 (br. s., 1H).

Example 23.

1-(4-(5-Chloro-6-(5-cyclopropyl-1H-indazol-4-yl)-7-fluoro-2,1-benzothiazol-3-yl)-1-piperazinyl)-2propen-1- He

Example 23 was prepared as described in Method 1 using (5-cyclopropyl-1H-indazol-4yl)boronic acid (see synthesis below) in step 7 and omitting step 8-3.

Mass/charge (ESI, +ve) 482.0 (M+H) ⁺ .

1H NMR (400 MHz, DMSO-d ₆ ) δ: 12.92-13.19 (1H, m), 8.02-8.21 (1H, m), 7.47-7.60 (2H, m ), 7.02-7.09 (1H, m), 6.80-6.93 (1H, m), 6.15-6.25 (1H, m), 5.71-5.82 (1H , m), 3.80-3.96 (4H, m), 3.60-3.72 (4H, m), 1.551.74 (1H, m), 0.72-0.79 (2H, m ), 0.58-0.71 (2H, m).

(5-Cyclopropyl-1H-indazol-4-yl)boronic acid he.c.

Br

tetrakis, 2 M Na ₂ CO _: CPME, 130 °C

Stage 1

LDA

Br

DMF/THF, -78 °C

Stage 2

dioxane, 100 °C Stage 4 ethylene glycol hydrazine hydrate, 150 °C Stage 3

Stage 1. 2-Bromo-1-cyclopropyl-4-fluorobenzene.

Add 2-bromo-4-fluoro-1iodobenzene (22 g, 73.1 mmol) and cyclopropylboronic acid (12.6 g, 146 mmol) in cyclopentyl methyl ether (1.1 L) to a 2-L round bottom flask at ambient temperature. . Na ₂ CO ₃ (2 M aq; 183 mL) was added and the reaction mixture was degassed with N2 gas for 20 minutes. Tetrakis (8.45 g, 7.31 mmol) was added and the reaction mixture was again degassed with N2 gas for 20 min. The reaction mixture was then transferred to a 5 L autoclave under N2 and heated to 130°C for 40 hours. The reaction mixture was cooled to ambient temperature, filtered through a pad of celite and washed with diethyl ether (200 ml). Water (500 ml) was added to the filtrate, and the organic layer was separated. The aqueous layer was extracted with diethyl ether (2x300 ml) and the combined organic layers were dried over anhydrous sodium sulfate and evaporated under reduced pressure. The crude material was adsorbed onto a silica gel plug and purified by chromatography (silica gel, 100% petroleum ether) to give 2-bromo-1-cyclopropyl-4-fluorobenzene.

GC-mS mass/charge: 214/216.

1H NMR (400 MHz, CDCl3) δ: 7.36-7.23 (m, 1H), 6.95 (dt, J=7.0, 1.5 Hz, 2H), 2.09 (ddd, J =13.8, 8.5, 5.4 Hz, 1H), 1.12-0.88 (m, 2H), 0.76-0.50 (m, 2H).

Stage 2. 2-Bromo-3-cyclopropyl-6-fluorobenzaldehyde.

2-Bromo-1-cyclopropyl-4-fluorobenzene (6.5 g, 30.2 mmol) in tetrahydrofuran (130 mL) under N2 was added to a 500 mL round bottom flask. LDA (18.1 mL, 36.3 mmol, 2 M in THF, 1.2 eq) was added dropwise at -78°C (internal temperature maintained at -65 to -70°C) and the reaction mixture was stirred for 1 hour. DMF (6 ml) was then added dropwise to the reaction mixture (internal temperature maintained at -65 to -70°C) and the reaction mixture was stirred for an additional 3 hours at -78°C. The reaction mixture was quenched with saturated aqueous ammonium chloride (100 ml) and slowly warmed to ambient temperature. The mixture was diluted with diethyl ether (200 ml) and the organic layer was separated and washed with brine (2x50 ml). The combined organic layers were dried over anhydrous Na ₂ SO ₄ and evaporated under reduced pressure. The crude material was adsorbed onto a silica gel plug and purified by chromatography (silica gel, 0-2% EtOAc/hexane) to give 2-bromo-3-cyclopropyl-6-fluorobenzal dehyde.

- 120 044387

GC-MS mass/charge: 242.

1H NMR (400 MHz, CDCl ₃ ) δ: 10.43 (d, J=1.5 Hz, 1H), 7.26-7.12 (m, 1H), 7.06 (t, J=9, 3 Hz, 1H), 2.15 (td, J=8.4, 4.3 Hz, 1H), 1.17-0.94 (m, 2H), 0.78-0.52 (m, 2H ).

Step 3. 4-Bromo-5-cyclopropyl-1H-indazole.

2-bromo-3-cyclopropyl-6-fluorobenzaldehyde (4 g, 16.5 mmol) and hydrazine hydrate (4.0 mL, 82 mmol) in ethylene glycol (40 mL) were added to a 100 mL sealed tube. The reaction mixture was stirred for 2 hours at 90°C and then heated to 150°C for 16 hours. The reaction mixture was cooled to ambient temperature and ice water (40 ml) and EtOAc (50 ml) were added. The organic layer was separated and the aqueous layer was extracted with EtOAc (2x40 ml). The combined organic layers were washed with water (2x40 ml) and brine (40 ml), dried over anhydrous aqueous sodium sulfate and concentrated in vacuo. The crude material was adsorbed onto a silica gel plug and purified by chromatography (silica gel, 0-20% EtOAc/hexane) to give 4bromo-5-cyclopropyl-1H-indazole. The compound was purified by preparative reverse phase liquid chromatography (YMC: C ₁₈ , 150x20 mm, 5 μm; mobile phase: 0.1% TFA in water and acetonitrile; flow rate: 15 ml/min.) to obtain the pure compound.

MS (ESI, positive ion) mass/charge: 237/239.0 (M+1).

1H NMR (400 MHz, DMSO-d6) δ: 13.31 (s, 1H), 7.97 (s, 1H), 7.46 (d, J=8.6 Hz, 1H), 6.97 ( d, J=8.6 Hz, 1H), 2.21 (tt, J=8.5, 5.3 Hz, 1H), 1.24-0.87 (m, 2H), 0.93-0 .33 (m, 2H).

Step 4. 5-Cyclopropyl-1H-indazol-4-yl)boronic acid.

4-Bromo-5-cyclopropyl-1H-indazole (0.62 g, 2.6 mmol) and bis-(pinacolato)diboron (0.996 g, 3.92 mmol) in 1,4- dioxane (25 ml); Potassium acetate (0.77 g, 7.84 mmol) was added and the reaction mixture was degassed with N2 gas for 10 min. PdCl ₂ (dppf) DCM adduct (0.213 g, 0.261 mmol) was added to the reaction mixture; the reaction mixture was again degassed with N2 gas for 10 minutes, then heated to 100°C for 16 hours. The reaction mixture was cooled to ambient temperature, filtered through a pad of celite and washed with EtOAc (50 ml). The filtrate was concentrated in vacuo and the crude material was adsorbed onto a silica gel plug and purified by chromatography (silica gel, 0-50% EtOAc/hexane). The compound was further purified by preparative reverse phase liquid chromatography (Grace column; 0-70% MeCN/water) to give 5-cyclopropyl-1Nindazol-4-yl)boronic acid. MS (ESI, positive ion) mass/charge: 285.2 (M+1).

1H NMR (400 MHz, DMSO-d6) δ: 12.88 (s, 1H), 8.13 (q, J=1.3 Hz, 1H), 7.50 (d, J=8.7 Hz, 1H), 6.83 (dd, J=8.8, 1.4 Hz, 1H), 2.78-2.60 (m, 1H), 1.38 (d, J=1.4 Hz, 12H ), 1.07-0.85 (m, 2H), 0.75-0.48 (m, 2H).

Example 24.

-(4-(5 -Chloro-7-fluoro-6-(3 -(methylamino)-1 -isoquinolinyl)-2,1 -benzothiazol-3 -yl)-1 -piperazinyl)-2propen-1-one

Step 1. 1-(5-Chloro-7-fluoro-3-(piperazin-1-yl)benzo[c]isothiazol-6-yl)-N-methylisoquinolin-3amine.

A solution of tert-butyl-4-(6-bromo-5-chloro-7-fluorobenzo[c]isothiazol-3-yl)piperazin-1-carboxylate (intermediate D, 30 mg, 0.067 mmol) in tetrahydrofuran (0. 6 ml) at 0°C, a solution of isopropylmagnesium chloride (2.0 M solution in tetrahydrofuran, 0.050 ml, 0.100 mmol) was added. The mixture was stirred for 5 min before zinc chloride (1.9 M solution in 2-methyltetrahydrofuran, 0.053 mL, 0.100 mmol) was added and the reaction mixture was warmed to room temperature. and stirred for 40 minutes. The reaction mixture was then transferred to a vial containing Sphos Pd G3 (5.76 mg, 6.66 µmol) and tert-butyl(1-bromoisoquinolin-3-yl)(methyl)carbamate (24.7 mg, 0.073 mmol, cm. synthesis below) and heated to 70°C overnight. The crude reaction mixture was diluted with our help. aq. a solution of NH ₄ Cl (50 ml) and EtOAc (50 ml). The organic layer was separated, dried over Na ₂ SO ₄ , filtered and concentrated. Purification by silica gel column chromatography eluting with 6-20% MeOH in DCM provided 1-(5-chloro-7-fluoro-3-(piperazin-1-yl)benzo[c]isothiazol-6yl)-N- methylisoquinoline-3-amine.

Mass/charge (ESI, +ve) 428.1 (M+H) ⁺ .

Synthesis of tert-butyl(1-bromoisoquinolin-3-yl)(methyl)carbamate.

A solution of 1-bromoisoquinoline-3-amine (200 mg, 0.897 mmol, Maybridge Chemical Co., Altrincham, UK) in tetrahydrofuran (5 ml) at RT. added sodium bis(trimethylsilyl)amide (1 M

- 121 044387 solution in tetrahydrofuran, 1.79 ml, 1.79 mmol). The mixture was stirred for 10 min before adding a solution of Boc anhydride (0.208 ml, 0.897 mmol) in THF (1 ml). The reaction mixture was stirred for 5 min before dilution with us. aq. NH4Cl solution (50 ml) and EtOAc (50 ml). The organic layer was separated, dried over Na ₂ SO ₄ , filtered and concentrated. Purification by silica gel column chromatography eluting with 0-20% EtOAc in heptane provided tert-butyl(1-bromoisoquinolin-3-yl)carbamate.

Mass/charge (ESI, +ve) 345.0 (M+Na) ⁺ .

Into a solution of tert-butyl(1-bromoisoquinolin-3-yl)carbamate (140 mg, 0.433 mmol) in tetrahydrofuran (3 ml) at room temperature. sodium hydride (60% dispersion in mineral oil, 22.52 mg, 0.563 mmol) was added. The mixture was stirred for 15 min before adding methyl iodide (0.033 ml, 0.520 mmol). After stirring overnight, the reaction mixture was diluted with our help. aq. NH4Cl solution (50 ml) and EtOAc (50 ml). The organic layer was separated, dried over Na ₂ SO ₄ , filtered and concentrated. Purification by silica gel column chromatography eluting with 0-10% EtOAc in heptane provided tert-butyl(1-bromoisoquinolin-3-yl)(methyl)carbamate.

¹ H NMR (400 MHz, METHANOL-d ₄ ) δ: 8.25 (d, J=8.61 Hz, 1H), 7.88-7.95 (m, 2H), 7.79 (t, J =7.5 Hz, 1H), 7.70 (t, J=7.6 Hz, 1H), 3.42 (s, 3H), 1.54 (s, 9H).

Mass/charge (ESI, +ve) 359.1 (M+H) ⁺ .

Step 2. 1-(4-(5-Chloro-7-fluoro-6-(3-(methylamino)-1-isoquinolinyl)-2,1-benzothiazol-3-yl)-1 piperazinyl)-2-propen- 1-he.

The procedure is similar to method 1, stage 8-2. Purify by silica gel column chromatography eluting with 0-14% MeOH in DCM over 15 min.

1H NMR (400 MHz, METHANOL-d ₄ ) δ: 7.90 (s, 1H), 7.63 (d, J=8.5 Hz, 1H), 7.43 (t, J=7.4 Hz , 1H), 7.24 (d, J=8.5 Hz, 1H), 7.05 (t, J=7.4 Hz, 1H), 6.72-6.84 (m, 1H), 6 .67 (s, 1H), 6.15-6.28 (m, 1H), 5.68-5.81 (m, 1H), 3.87-3.97 (m, 4H), 3.63 (m, 4H), 2.90 (s, 3H).

Mass/charge (ESI, +ve) 482.0 (M+H) ⁺ .

Example 25.

-(4-(6-(3 -Amino-1 -isoquinolinyl)-5 -chloro-7-fluoro-2,1 -benzothiazol-3 -yl)-1 -piperazinyl)-2-propen1-one

Step 1. tert-Butyl-4-(6-(3-((tert-butoxycarbonyl)amino)isoquinolin-1-yl)-5-chloro-7-fluorobenzo[c]isothiazol-3-yl)piperazin-1- carboxylate.

The procedure is similar to example 25, step 1, using a solution of 1.3 M isopropylmagnesium chloride in THF instead of a solution of isopropylmagnesium chloride and bis-(2-methyl-2-propanyl) (1-bromo-3isoquinolinyl)-2-imidodicarbonate (synthesis below) instead tert-butyl(1-bromoisoquinolin-3-yl)(methyl)carbamate.

Mass/charge (ESI, +ve) 614.2 (M+H) ⁺ .

Synthesis of bis-(2-methyl-2-propanyl)(1-bromo-3-isoquinolinyl)-2-imidodicarbonate.

Boc anhydride (3.12 mL, 13 .45 mmol) and DMAP (0.055 g, 0.448 mmol). The reaction mixture was heated to room temperature. and stirred overnight. The reaction mixture was then diluted with our help. aq. NH4Cl solution (100 ml) and DCM (50 ml). The organic layer was separated, dried over Na ₂ SO ₄ , filtered and concentrated. Purification by silica gel column chromatography eluting with 0-10% EtOAc in heptane over 15 minutes provided bis-(2-methyl-2-propanyl)(1-bromo-3-isoquinolinyl)-2-imidodicarbonate.

1H NMR (400 MHz, METHANOL-d ₄ ) δ: 8.36 (d, J=8.5 Hz, 1H), 8.03 (d, J=8.1 Hz, 1H), 7.77-7 .92 (m, 3H), 1.44 (s, 18H).

Mass/charge (ESI, +ve) 267.0 (M+H) ⁺ .

Step 2. 1-(4-(6-(3-Amino-1-isoquinolinyl)-5-chloro-7-fluoro-2,1-benzothiazol-3-yl)-1-piperazinyl)2-propen-1- He.

The procedure is similar to Method 1, Steps 8-1 and 8-2, but in Step 8-1 TFA in DCM was used instead of 4 M HCl in dioxane/MeOH. Purify by silica gel column chromatography eluting with 0-12% MeOH in DCM. This material was then subjected to SFC purification: diol column (21.2x250 mm, 5 µm) using 17% (20 mM NH ₃ in MeOH) in supercritical CO2 (total flow rate was 7 g/min), yielding 1- (4-(6-(3-amino-1-isoquinolinyl)-5-chloro-7fluoro-2,1-benzothiazol-3-yl)-1-piperazinyl)-2-propen-1-one.

- 122 044387

1H NMR (400 MHz, METHANOL^) δ: 7.85 (s, 1H), 7.53 (d, J=8.5 Hz, 1H), 7.39 (t, J=7.6 Hz, 1H ), 7.23 (d,

J=8.5 Hz, 1H), 7.03 (t, J=7.8 Hz, 1H), 6.82 (s, 1H), 6.71 (dd, J=10.8, 16.8 Hz, 1H), 6.2 (dd, J=1.5, 16.8 Hz, 1H),

5.70 (dd, J=1.5, 10.8 Hz, 1H), 3.82-3.93 (m, 4H), 3.50-3.66 (m, 4H).

Mass/charge (ESI, +ve) 468.0 (M+H)+.

Example 26.

1-(4-(6-(2-Amino-4-quinolinyl)-5-chloro-7-fluoro-2,1-benzothiazol-3-yl)-1-piperazinyl)-2-propen-1-one

Stage 1. carboxylate.

Solution (intermediate tert-Butyl-4-(5-chloro-7-fluoro-6-(tributylstannyl)benzo[c]isothiazol-3-yl)piperazin-1 tert-butyl-4-(6-bromo-5-chloro- 7-fluorobenzo[c]isothiazol-3-yl)piperazin-1-carboxylate compound D, 320 mg, 0.710 mmol), 1,1,1,2,2,2-hexabutyldistannan (824 mg,

1.420 mmol) and tetrakis-(triphenylphosphine) palladium(0) (82 mg, 0.071 mmol, Strem Chemicals Inc., Newburyport, MA, USA) in N,N-dimethylacetamide (5 ml) were heated in a sealed vial by microwave at 160 °C for 40 min. The reaction mixture was diluted for us. aq. NaHCO ₃ solution (50 ml), saline (50 ml) and EtOAc (100 ml). The organic layer was separated, dried over Na ₂ SO ₄ , filtered and concentrated. Purification by silica gel column chromatography eluting with 0-30% EtOAc in heptane provided tert-butyl-4-(5-chloro-7-fluoro-6-(tributylstannyl)benzo[c]isothiazol-3-yl)piperazine-1 -carboxylate.

Mass/charge (ESI, +ve) 662.2 (M+H)+.

Step 2. 2-Methyl-2-propanyl-4-(6-(2-(bis-(((2-methyl-2-propanyl)oxy)carbonyl)amino)-4quinolinyl)-5-chloro-7-fluoro -2, 1-benzothiazol-3-yl)-1-piperazinecarboxylate.

Di-tert-butyl-(4-bromoquinolin-2-yl)-2-imidodicarbonate solution (19.2 mg, 0.045 mmol, prepared analogously to bis-(2-methyl-2-propanyl)(1-bromo-3-isoquinolinyl) )-2-imidodicarbonate from Example 26 using 4-bromoquinoline-2-amine (Ark Pharm Inc. Arlington Heights, IL, USA) as starting material, tert-butyl-4-(5-chloro-7-fluoro- 6-(tributylstannyl)benzo[c]isothiazol-3yl)piperazine-1-carboxylate (20 mg, 0.030 mmol), tetrakis-(triphenylphosphine)palladium(0) (6.99 mg, 6.05 µmol, Strem Chemicals Inc. , Newburyport, MA, USA), copper(I) iodide (1.153 mg, 6.05 µmol) and cesium fluoride (13.79 mg, 0.091 mmol) in DMF (0.5 ml) were heated in a sealed vial at 60°C for 30 min. The crude reaction mixture was diluted with aqueous saline solution of NaHCO ₃ (50 ml) and EtOAc (100 ml). The organic layer was separated, dried over Na ₂ SO ₄ , filtered and concentrated. Purification by column chromatography on silica gel eluting using 0-50% EtOAc in heptane provided 2-methyl-2-propanyl 4-(6-(2-(bis-(((2-methyl-2propanyl)oxy)carbonyl)amino)-4-quinolinyl)- 5-chloro-7-fluoro-2,1-benzothiazol-3-yl)-1-piperazinecarboxylate.

Mass/charge (ESI, +ve) 714.2 (M+H)+.

Step 3. 1-(4-(6-(2-Amino-4-quinolinyl)-5-chloro-7-fluoro-2,1-benzothiazol-3-yl)-1-piperazinyl)-2propen-1-one .

The procedure is similar to Method 1, Steps 8-1 and 8-2, but in Step 8-1 TFA in DCM was used instead of 4 M HCl in dioxane/MeOH.

1H NMR (400 MHz, METHANOL^) δ: 7.90 (s, 1H), 7.53 (d, J=8.2 Hz, 1H), 7.42-7.49 (m, 1H), 7 ,10 (d, J=8.0 Hz, 1H), 7.03-7.08 (m, J=7.6 Hz, 1H), 6.67-6.81 (m, 2H), 6, 19 (dd,J=1.8, 16.6 Hz, 1H), 5.72 (dd, J=1.8, 10.6 Hz, 1H), 3.87-3.93 (m, 4H) , 3.56-3.66 (m, 4H).

Mass/charge (ESI, +ve) 468.0 (M+H)+.

- 123 044387

Example 27.

1-(4-(3-(2-Fluoro-6-hydroxyphenyl)-2-methyl-5-(2-(2-pronanyl)phenyl)nirido[2,3-d]pyridazin-8-yl)1 - piperazinyl)-2-propen-1-one

Step 1. 6,7-Dihydropyrido[2,3-th]pyridazine-5,8-dione.

Hydrazine (1.26 ml, 40.2 mmol) was added to a stirred solution of 2,3-pyridinedicarboxylic anhydride (4.00 g, 26.8 mmol) in ethanol (100 ml). The reaction mixture was refluxed for 16 hours before cooling to room temperature. and concentrated in vacuo to give crude 6,7-dihydropyrido[2,3-d]pyridazine-5,8-dione, which was used directly in the next step.

Mass/charge (ESI) M+H: 164.1.

Step 2. 5,8-Dichloropyrido[2,3-d]pyridazine.

Pyridine (4.57 ml, 53.7 mmol) was added to a mixture of crude 6,7-dihydropyrido[2,3d]pyridazine-5,8-dione (4.38 g, 26.8 mmol) in phosphorus(U) oxychloride (20.1 ml, 215 mmol). The reaction mixture was stirred at 100°C for 2 hours. The reaction mixture was cooled and slowly poured into rapidly stirred water (250 ml) at ~10°C. The aqueous suspension was stirred for 15 min before extraction with EtOAc (250 ml).

The organic layer was separated, washed with brine (200 ml), dried over MgSO ₄ , filtered and concentrated in vacuo. Chromatographic purification of the residue (silica gel, 0 to 100% EtOAc in heptane) provided 5,8-dichloropyrido[2,3-0]pyridazine.

Ή NMR (400 MHz, CHLOROFORM-d) δ: 9.41 (1H, dd, J=4.30, 1.56 Hz), 8.65 (1H, dd, J=8.41, 1.56 Hz ), 8.02 (1H, dd, J=8.41, 4.30 Hz).

Mass/charge (ESI) M+H: 200.0.

Step 3. 3,5-Dichloropyrido[2,3-d]pyridazin-8(7H)-one and 3,8-dichloropyrido[2,3-d]niridazin-5(6H)-one.

N-Chlorosuccinimide (1268 mg, 9.50 mmol, TCI America, Portland, OR, USA) was added to a stirred solution of 5,8-dichloropyrido[2,3-d]pyridazine (950 mg, 4.75 mmol) in acetic acid (20 ml) and the reaction mixture was heated to 100°C for 16 hours. Additional N-chlorosuccinimide (1268 mg, 9.50 mmol, TCI America, Portland, OR, USA) was added and the reaction mixture was stirred at 100°C in for an additional 4 hours. Additional N-chlorosuccinimide (634 mg, 4.75 mmol, TCI America, Portland, OR, USA) was added and the reaction mixture was stirred for an additional 4 hours. The reaction mixture was then diluted with water (75 ml) and extracted three times with EtOAc (100 ml). The combined organic layers were washed with brine (150 ml), dried over MgSO ₄ , filtered and concentrated in vacuo. Chromatographic purification of the residue (silica gel, 0 to 75% EtOAc in heptane) provided a regioisomeric mixture of 3,5-dichloropyrido[2,3-d]pyridazin-8(7H)-one compound and 3,8-dichloropyrido[2,3 -d]pyridazin5(6H)-one.

Mass/charge (ESI) M+H: 215.9.

Step 4. 3,5,8-Trichloropyrido[2,3-d]pyridazine.

Pyridine (2.024 ml, 23.79 mmol) was added to a regioisomeric mixture of 3,5-dichloropyrido[2,3d]pyridazin-8(7H)-one and 3,8-dichloropyrido[2,3-d]pyridazin-5(6H )-one (2.57 g, 11.90 mmol) in phosphorus oxychloride (8.90 ml, 95 mmol). The reaction mixture was stirred at 100°C for 1.5 hours. The reaction mixture was cooled and slowly poured into rapidly stirred water (150 ml) at ~10°C. The aqueous suspension was stirred for 15 min before extraction with EtOAc (200 ml). The organic layer was separated, washed with brine (150 ml), dried over MgSO ₄ , filtered and concentrated in vacuo. Chromatographic purification of the residue (silica gel, 0 to 50% EtOAc in heptane) provided 3,5,8-trichloropyrido[2,3-d]pyridazine.

1H NMR (400 MHz, CHLOROFORM-d) δ: 9.27 (1H, d, J=2.35 Hz), 8.58 (1H, d, J=2.35 Hz).

Mass/charge (ESI) M+H: 233.9.

Step 5. tert-Butyl 4-(3,5-dichloropyrido[2,3-d]pyridazin-8-yl)piperazine-1-carboxylate.

1-Boc-piperazine (278 mg, 1.494 mmol) was added to a stirred mixture of 3,5,8-trichloropyrido[2,3-d]pyridazine (292 mg, 1.245 mmol) and triethylamine (0.350 ml, 2.491 mmol) in dimethyl sulfoxide ( 5 ml). The reaction mixture was stirred at room temperature. for 3 hours before diluting with EtOAc (75 ml) and washing with saturated aqueous sodium bicarbonate (75 ml).

The organic layer was separated, washed with brine (50 ml), dried over MgSO4, filtered and concentrated in vacuo. Chromatographic purification of the residue (silica gel, 0 to 25% acetone in heptane) provided tert-butyl 4-(3,5-dichloropyrido[2,3d]pyridazin-8-yl)piperazin-1-carboxylate, the first of two eluted regioisomers.

1H NMR (400 MHz, CHLOROFORM-d) δ: 9.01 (1H, d, J=2.54 Hz), 8.43 (1H, d, J=2.54 Hz), 4.04-4, 15 (4H, m), 3.64-3.70 (4H, m), 1.50 (9H, s).

Mass/charge (ESI) M+H: 384.0.

Step 6. tert-Butyl-4-(3-chloro-5-(2-isopropylphenyl)pyrido[2,3-d]pyridazin-8-yl)piphenrazin-1carboxylate.

tert-Butyl-4-(3,5-dichloropyrido[2,3-d]pyridazin-8-yl)piperazine-1-carboxylate (199 mg, 0.518 mmol), 2-isopropylphenylboronic acid (93 mg, 0.570 mmol, Alfa Aesar, Haver Hill, MA, USA), tetrakis(triphenylphosphine) palladium (59.8 mg, 0.052 mmol, Strem Chemicals Inc., Newburyport, MA, USA) and sodium carbonate (2 M aq, 1.036 ml, 2.072 mmol) mixed in 1,4-dioxane (4 ml) under argon. The reaction mixture was stirred at 40°C for 16 hours. The reaction mixture was cooled to room temperature, diluted with EtOAc (50 ml) and washed with water (40 ml). The organic layer was separated, washed with brine (50 ml), dried over MgSO ₄ , filtered and concentrated in vacuo. Chromatographic purification of the residue (silica gel, 0 to 50% EtOAc in heptane) provided a mixture of starting material and desired product. The mixture was resubjected to the original reaction conditions using a smaller amount of 2-isopropylphenylboronic acid (56 mg, 0.342 mmol, Alfa Aesar, Haver Hill, MA, USA). The mixture was stirred at 40°C for 16 hours. Additional 2-isopropylphenylboronic acid (28 mg, 0.171 mmol, Alfa Aesar, Haver Hill, MA, USA) was added and the reaction mixture was stirred for a further 6 hours. The reaction mixture was cooled to .t., diluted with EtOAc (50 ml) and washed with water (40 ml). The organic layer was separated, washed with brine (50 ml), dried over MgSO4, filtered and concentrated in vacuo. Chromatographic purification of the residue (silica gel, 0 to 50% EtOAc in heptane) provided tert-butyl-4-(3-chloro-5-(2-isopropylphenyl)pyrido[2,3-d]pyridazin-8-yl)piperazine -1carboxylate.

1H NMR (400 MHz, CHLOROFORM-d) δ: 8.95 (1H, d, J=2.35 Hz), 7.72 (1H, d, J=2.54 Hz), 7.45-7, 53 (2H, m), 7.26-7.33 (1H, m), 7.16-7.21 (1H, m) 4.04-4.23 (4H, m) 3.66-3, 73 (4H, m) 2.67 (1H, spt, J=6.75 Hz) 1.48 (9H, s) 1.16 (3H, d, J=6.85 Hz) 1.03 (3H, d, J=6.85 Hz).

Mass/charge (ESI) M+H: 468.2.

Step 7. tert-Butyl-4-(3-chloro-5-(2-isopropylphenyl)-2-methylpyrido[2,3-d]pyridazin-8yl)piperazine-1-carboxylate.

Methyllithium (1.6 M diethyl ether solution, 0.137 mL, 0.219 mmol) was added to the stirred solution of tert-butyl-4-(3-chloro-5-(2-isopropylphenyl)pyrido[2,3-d]pyridazin-8 -yl)piperazine-1carboxylate (93 mg, 0.199 mmol) in tetrahydrofuran (1 ml) at -78°C.

- 125 044387

The reaction mixture was stirred at -78°C for 5 minutes before warming to 0°C and stirring for 30 minutes. The reaction mixture was again cooled to -78°C and additional methyl lithium (1.6 M solution in diethyl ether, 0.068 ml, 0.109 mmol) was added. The reaction mixture was stirred at -78°C for 5 minutes before warming to 0°C and stirred for a further 15 minutes. The reaction mixture was quenched with water (20 ml) and extracted with EtOAc (30 ml). The organic layer was separated, washed with brine (20 ml), dried over MgSO ₄ , filtered and concentrated in vacuo to give crude tert-butyl-4-(3-chloro-5-(2-isopropylphenyl)-2-methyl- 1,2-dihydropyrido[2,3-d]pyridazin8-yl)piperazine-1-carboxylate.

Mass/charge (ESI) M+H: 484.3.

4,5-Dichloro-3,6-dioxo-1,4-cyclohexadiene-1,2-dicarbonitrile (45.0 mg, 0.198 mmol) was added to the stirred mixture of crude tert-butyl-4-(3-chloro-5- (2-isopropylphenyl)-2-methyl-1,2dihydropyrido[2,3-d]pyridazin-8-yl)piperazine-1-carboxylate (96 mg, 0.198 mmol) in dichloromethane (2 ml). The reaction mixture was stirred at room temperature. within 10 min. The reaction mixture was diluted with DCM (30 ml) and washed with water (20 ml). The organic layer was separated, dried over MgSO ₄ , filtered and concentrated in vacuo. Chromatographic purification of the residue (silica gel, 0 to 50% EtOAc in heptane) provided tert-butyl-4-(3-chloro-5-(2isopropylphenyl)-2-methylpyrido[2,3-d]pyridazin-8-yl) piperazine-1-carboxylate.

^1H NMR (400 MHz, CHLOROFORM-d) δ: 7.72 (1H, s), 7.51-7.55 (2H, m), 7.32-7.37 (1H, m), 7, 22-7.27 (1H, m) 4.08-4.25 (4H, m) 3.71-3.79 (4H, m) 2.87 (3H, s) 2.73 (1H, spt, J=6.68 Hz) 1.54 (9H, s) 1.21 (3H, d, J=6.85 Hz) 1.07 (3H, d, J=6.85 Hz).

Mass/charge (ESI) M+H: 482.1.

Step 8. tert-Butyl-4-(3-(2-fluoro-6-hydroxyphenyl)-5-(2-isopropylphenyl)-2-methylpyrido[2,3b]pyridazin-8-yl)piperazine-1-carboxylate.

tert-Butyl-4-(3-chloro-5-(2-isopropylphenyl)-2-methylpyrido[2,3-b]pyridazin-8-yl)piperazine-1carboxylate (78 mg, 0.162 mmol), (2-fluoro -6-Hydroxyphenyl)boronic acid (101 mg, 0.647 mmol, Combi-Blocks), Sphos Pd G3 (14.00 mg, 0.016 mmol) and sodium carbonate (2 M aq., 0.324 mL, 0.647 mmol) were mixed in 1.2 -dimethoxyethane (1 ml) under argon and then heated at 80°C for 2.5 hours. The reaction mixture was cooled, diluted with EtOAc (30 ml) and washed with water (25 ml). The organic layer was separated, washed with brine (25 ml), dried over MgSO ₄ , filtered and concentrated in vacuo. Chromatographic purification of the residue (silica gel, 0 to 50% EtOAc in heptane) provided tert-butyl-4-(3-(2-fluoro-6hydroxyphenyl)-5-(2-isopropylphenyl)-2-methylpyrido[2,3- b]pyridazin-8-yl)piperazine-1-carboxylate (66 mg, 0.118 mmol, 73.1% yield).

Mass/charge (ESI) M+H: 558.2.

Step 9. 3-Fluoro-2-(5-(2-isopropylphenyl)-2-methyl-8-(piperazin-1-yl)pyrido[2,3-b]pyridazin-3yl)phenol.

Trifluoroacetic acid (0.2 mL, 2.68 mmol) was added to the stirred solution of tert-butyl-4-(3-(2-fluoro-6-hydroxyphenyl)-5-(2-isopropylphenyl)-2-methylpyrido[2,3- d]pyridazin-8yl)piperazine-1-carboxylate (64 mg, 0.115 mmol) in dichloromethane (0.5 ml). The reaction mixture was stirred at room temperature. within 30 min. The reaction mixture was diluted with DCM (30 ml) and quenched with saturated aqueous sodium bicarbonate (20 ml). The organic layer was separated, dried over MgSO ₄ , filtered and concentrated in vacuo to give crude 3-fluoro-2-(5-(2-isopropylphenyl)-2-methyl-8-(piperazin-1-yl)pyrido[2,3 -d]pyridazin-3-yl)phenol.

Mass/charge (ESI) M+H: 458.1.

Step 10 -2-en-1-on.

Acryloyl chloride (9.45 μL, 0.116 mmol) was added to the stirred mixture of 3-fluoro-2-(5-(2isopropylphenyl)-2-methyl-8-(piperazin-1-yl)pyrido[2,3-d]pyridazin- 3-yl)phenol (53 mg, 0.116 mmol) and triethylamine (0.049 ml, 0.348 mmol) in dichloromethane (1 ml) at 0°C. The reaction mixture was stirred at 0°C for 10 minutes. The reaction mixture was diluted with DCM (25 ml) and quenched with saturated aqueous sodium bicarbonate (20 ml). The organic layer was separated, dried over MgSO ₄ , filtered and concentrated in vacuo. Chromatographic purification of the residue (silica gel, 0 to 100% EtOAc in heptane) provided 1-(4-(3-(2-fluoro-6hydroxyphenyl)-2-methyl-5-(2-(2-propanyl)phenyl)pyrido [2,3-d]pyridazin-8-yl)-1-piperazinyl)-2propen-1-one.

1H NMR (400 MHz, CHLOROFORM-d) δ: 9.51 (0.6H, br s), 8.98 (0.4H, br s), 7.63 (0.4H, s), 7.58 (0.6H, s), 7.35-7.43 (2H, m), 7.10-7.26 (3H, m), 6.78 (1H, dd, J=16.63, 8, 22 Hz), 6.59-6.71 (2H, m), 6.36 (1H, dd, J=16.82, 1.57 Hz), 5.78 (1H, dd, J=10.56 , 1.37 Hz), 4.10-4.38 (4H, m), 3.80-4.03 (4H, m), 2.60-2.72 (1H, m), 2.61 ( 1.2H, s), 2.59 (1.8H, s), 0.91-1.08 (6H, m).

Mass/charge (ESI) M+H: 512.3.

- 126 044387

Example 28.

-(4-(7-Chloro-6-(2-fluoro-6-hydroxyphenyl)-4-((1R)-1-phenylethyl)-1-phthalazinyl)-1-piperazinyl)-2propen-1-one and 1 -(4-(7-chloro-6-(2-fluoro-6-hydroxyphenyl)-4-((1S)-1-phenylethyl)-1-phthalazinyl)-1 piperazinyl)-2-propen-1-one

A mixture of α-methylbenzylzinc bromide (0.5 M in THF, 492 µl, 0.246 mmol), tetrakis(triphenylphosphine)palladium (5.68 mg, 4.92 µmol, Strem Chemicals Inc., Newburyport, MA, USA) and 1 - (4-(4,7-dichloro-6-(2-fluoro-6-hydroxyphenyl)phthalazin-1 -yl)piperazin-1 -yl)prop-2-en-1-one (intermediate I, 22 mg, 0.049 mmol) was stirred at 60°C in a sealed vial for 16 hours. The reaction mixture was concentrated, and chromatographic purification of the residue (silica gel, 0 to 100% EtOAc in heptane) provided a mixture of 1-(4-(7-chloro-6 -(2-fluoro-6hydroxyphenyl)-4-(( 1R)-1-phenylethyl)-1-phthalazinyl)-1-piperazinyl)-2-propen-1-one and 1 -(4-(7-chloro-6 -(2fluoro-6-hydroxyphenyl)-4-((1S)-1-phenylethyl)-1-phthalazinyl)-1-piperazinyl)-2-propen-1-one.

1H NMR (400 MHz, METHANOL) δ: 8.27 (1H, s), 8.15 (0.33H, s), 8.10 (0.67 H, s), 7.19-7.31 ( 5H, m), 7.10-7.16 (1H, m), 6.86 (1H, dd, J=16.73, 10.66 Hz), 6.62-6.78 (2H, m) , 6.27 (1H, dd, J=16.82, 1.96 Hz), 5.80 (1H, dd, J=10.66, 1.86 Hz), 4.94-5.01 (1H , m), 3.93-4.03 (4H, m), 3.49-3.60 (4H, m), 1.81 (3H, d, J=7.04 Hz).

Mass/charge (ESI) M+H: 517.1.

Example 29.

1-(4-(7-Chloro-4-(4-fluorobenzyl)-6-(2-fluoro-6-hydroxyphenyl)-1-phthalazinyl)-1-piperazinyl)-2propen-1-one

4-Fluorobenzylzinc chloride (0.5 M in THF, 0.089 mL, 0.044 mmol) was added to the stirred mixture of 1-(4-(4,7-dichloro-6-(2-fluoro-6-hydroxyphenyl)phthalazin-1-yl )piperazin-1-yl)prop-2-en-1-one (intermediate I, 18 mg, 0.040 mmol) and tetrakis-(triphenylphosphine) palladium (4.65 mg, 4.02 μmol, Strem Chemicals Inc., Newburyport, Massachusetts, USA) in tetrahydrofuran (0.1 ml) in a sealed vial under argon. The reaction mixture was stirred at room temperature. for 2 hours before heating to 40°C for 3 hours. Additional 4-fluorobenzylzinc chloride (0.089 mL, 0.044 mmol) was added and the reaction mixture was stirred at 40°C for an additional 16 hours. Additional 4-fluorobenzylzinc chloride was added (0.089 ml, 0.044 mmol) and the reaction mixture was heated to 60°C and stirred for 6 hours. The reaction mixture was concentrated in vacuo. Chromatographic purification of the residue (silica gel, 0 to 100% EtOAc in heptane) provided 1-(4-(7-chloro-4-(4-fluorobenzyl)-6-(2-fluoro-6-hydroxyphenyl)-1-phthalazinyl )-1piperazinyl)-2-propen-1-one.

^1H NMR (400 MHz, METHANOL) δ: 8.32 (1H, s) 8.19 (1H, s), 7.26-7.34 (3H, m) 6.98 (2H, t, J= 8.71 Hz) 6.69-6.91 (3H, m) 6.28 (1H, dd, J=16.92, 1.86 Hz) 5.82 (1H, dd, J=1056, 1, 76 Hz) 4.54-4.65 (2H, m) 3.99 (4H, m) 3.58 (4H, m).

Mass/charge (ESI) M+H: 521.2.

- 127 044387

Examples 30 and 31.

2-(1-(4-Acryloyl-1-piperazinyl)-7-chloro-4-phenyl-6-phthalazinyl)-3-fluorophenol (Example 30) and 2-(4-(4-acryloyl-1-piperazinyl) -7-chloro-1-phenyl-6-phthalazinyl)-3-fluorophenol (example 31)

Step 1. 1,4,6,7-tetrachlorophthalazine (intermediate L). Pyridine (431 μL, 5.28 mmol) was added to a stirred mixture of 6,7-dichloro-2,3-dihydrophthalazine-1,4-dione (intermediate G, 610 mg, 2.64 mmol) in phosphorus oxychloride (2, 4 ml, 26.4 mmol). The reaction mixture was heated to 100°C for 2 hours, then cooled and slowly poured into rapidly stirred water (75 ml) at ~10°C. The resulting suspension was filtered and the solid was washed with water to obtain 1,4,6,7-tetrachlorophthalazine.

1H NMR (400 MHz, CHLOROFORM-d) δ: 8.43 (2H, s).

Mass/charge (ESI) M+H: 266.9.

Step 2. tert-Butyl 4-(4,6,7-trichlorophthalazin-1-yl)piperazine-1-carboxylate (intermediate M).

1-Boc-piperazine (340 mg, 1.824 mmol) was added to a stirred mixture of 1,4,6,7-tetrachlorophthalazine (intermediate L, 543 mg, 2.027 mmol) and triethylamine (0.846 ml, 6.08 mmol) in dichloromethane ( 8 ml). The reaction mixture was stirred at room temperature. within 2 days. Additional 1-boc-piperazine (340 mg, 1.824 mmol) was added and the reaction mixture was stirred at room temperature. for another 23 hours. The reaction mixture was quenched with saturated aqueous sodium bicarbonate (20 ml) and extracted with DCM (30 ml).

The organic layer was separated, washed with brine (20 ml), dried over MgSO ₄ , filtered and concentrated in vacuo. Chromatographic purification of the residue (silica gel, 0 to 50% EtOAc in heptane) provided tert-butyl 4-(4,6,7-trichlorophthalazin-1yl)piperazine-1-carboxylate.

1H NMR (400 MHz, Chloroform-d) δ: 8.35 (1H, s), 8.12 (1H, s), 3.68-3.75 (4H, m), 3.45-3.52 (4H, m), 1.51 (9H, s).

Mass/charge (ESI) M+H: 417.0.

Step 3. tert-Butyl 4-(6,7-dichloro-4-phenylphthalazin-1-yl)piperazine-1-carboxylate.

tert-Butyl-4-(4,6,7-trichlorophthalazin-1-yl)piperazine-1-carboxylate (intermediate M, 95 mg, 0.227 mmol), tetrakis-(triphenylphosphine)palladium (26.3 mg, 0.023 mmol , Strem Chemicals Inc., Newburyport, MA, USA), phenylboronic acid (27.7 mg, 0.227 mmol) and sodium carbonate (2 M aq., 0.341 mL, 0.682 mmol) were mixed in 1,4-dioxane (1 mL) in sealed vial under argon atmosphere. The reaction mixture was stirred at 40°C for 24 hours. Additional tetrakis(triphenylphosphine)palladium (26.3 mg, 0.023 mmol) and phenylboronic acid (13.5 mg, 0.113 mmol) were added and the reaction mixture was stirred at 40°C for another 24 hours. The reaction mixture was quenched with saturated aqueous sodium bicarbonate (20 ml) and extracted with EtOAc (25 ml). The organic layer was separated, washed with brine (20 ml), dried over MgSO ₄ , filtered and concentrated in vacuo. Chromatographic purification of the residue (silica gel, 0 to 100% EtOAc in heptane) provided tert-butyl-4-(6,7

- 128 044387 dichloro-4-phenylphthalazin-1-yl)piperazine-1-carboxylate.

1H NMR (400 MHz, Chloroform-d), δ: 8.13 (1H, s), 8.07 (1H, s), 7.62-7.67 (2H, m), 7.50-7, 55 (3H, m), 3.65-3.74 (4H, m), 3.44-3.53 (4H, m), 1.47 (9H, s).

Mass/charge (ESI) M+H: 459.1.

Step 4. 6,7-Dichloro-1-phenyl-4-(piperazin-1-yl)phthalazine.

tert-Butyl-4-(6,7-dichloro-4-phenylphthalazin-1-yl)piperazine-1-carboxylate (68 mg, 0.148 mmol) was stirred in trifluoroacetic acid (1 ml, 13.46 mmol) at RT . within 20 min. The reaction mixture was quenched with saturated aqueous sodium bicarbonate (20 ml) and extracted twice with DCM (25 ml). The organic layer was separated, dried over MgSO ₄ , filtered and concentrated in vacuo to give crude 6,7-dichloro-1-phenyl-4-(piperazin-1yl)phthalazine, which was used directly in the next step.

Mass/charge (ESI) M+H: 359.0.

Step 5. 1 -(4-(6,7-Dichloro-4-phenylphthalazin-1 -yl)piperazin-1 -yl)prop-2-en-1-one.

Acryloyl chloride (0.013 mL, 0.162 mmol) was added to a stirred mixture of 6,7-dichloro-1-phenyl4-(piperazin-1-yl)phthalazine (53 mg, 0.148 mmol) and triethylamine (0.062 mL, 0.443 mmol) in dichloromethane (1 ml). The reaction mixture was stirred at room temperature. within 30 min. The reaction mixture was quenched with saturated aqueous sodium bicarbonate (15 ml) and extracted with DCM (20 ml). The organic layer was separated, dried over MgSO ₄ , filtered and concentrated in vacuo. Chromatographic purification of the residue (silica gel, 0 to 100% EtOAc in heptane) provided 1-(4-(6,7-dichloro-4-phenylphthalazin-1-yl)piperazin-1-yl)prop-2-ene-1 -she.

^1H NMR (400 MHz, Chloroform-d) δ: 8.20 (1H, s), 8.14 (1H, s), 7.66-7.75 (2H, m), 7.54-7, 62 (3H, m), 6.66 (1H, dd, J=16.63, 10.37 Hz), 6.37 (1H, dd, J=16.82, 1.96 Hz), 5.78 (1H, dd, J=10.56, 1.96 Hz), 3.85-4.04 (1H, m), 3.53-3.72 (1H, m).

Mass/charge (ESI) M+H: 431.2.

Step 6. 2-(1-(4-Acryloyl-1-piperazinyl)-7-chloro-4-phenyl-6-phthalazinyl)-3-fluorophenol and 2-(4-(4acryloyl-1-piperazinyl)-7- chloro-1-phenyl-6-phthalazinyl)-3-fluorophenol.

1-(4-(6,7-Dichloro-4-phenylphthalazin-1-yl)piperazin-1-yl)prop-2-en-1-one (43 mg, 0.104 mmol), 2-fluoro-6-hydroxyphenylboronic acid (17.84 mg, 0.114 mmol, Combi-Blocks Inc., San Diego, California, USA), Sphos Pd G3 (9.00 mg, 10.40 μmol) and sodium carbonate (2 M aq., 0.156 ml, 0.312 mmol) was mixed in 1,2-dimethoxyethane (0.5 ml) in a sealed vial under argon. The reaction mixture was stirred at 60°C for 3 hours. Additional 2-fluoro-6-hydroxyphenylboronic acid (8.92 mg, 0.057 mmol, Combi-Blocks Inc., San Diego, CA, USA) and SPhos Pd G3 were added (9.00 mg, 10.40 μmol) and the reaction mixture was stirred at 60°C for an additional 2 hours. The reaction mixture was quenched with saturated aqueous sodium bicarbonate (15 ml) and extracted with EtOAc (20 ml). The organic layer was separated, washed with brine (10 ml), dried over MgSO ₄ , filtered and concentrated in vacuo. Chromatographic purification of the residue (silica gel, 0 to 100% EtOAc in heptane) provided a mixture of two regioisomeric products. Preparative reverse phase chromatography (XBridge Prep C18 5 µm OBD, 150x30 mm; 35 to 55% (0.1% TFA in water) in (0.1% TFA in acetonitrile); flow=30 ml/min.) provided obtaining separated regioisomeric products. Fractions containing individual regioisomers were neutralized with saturated aqueous sodium bicarbonate and extracted with DCM, and the organic extracts were concentrated in vacuo. The separated regioisomers were further individually purified by column chromatography (silica gel, 0 to 100% EtOAc in heptane). 2-(1-(4-Acryloyl-1-piperazinyl)-7-chloro-4phenyl-6-phthalazinyl)-3-fluorophenol (Example 30) was the first elutable regioisomer obtained from preparative reverse phase chromatography.

1H NMR (400 MHz, CHLOROFORM-d), δ: 8.21 (1H, s), 8.06 (1H, s), 7.62-7.69 (2H, m), 7.45-7, 51 (3H, m), 7.24-7.32 (1H, m), 6.81-6.90 (1H, m), 6.75 (1H, t, J=8.41 Hz), 6 .65 (1H, dd, J=16.82, 10.56 Hz), 6.38 (1H, dd, J=16.82, 1.76 Hz), 5.79 (1H, dd, J=10 .56, 1.76 Hz), 3.86-4.02 (4H, m), 3.57-3.76 (4H, m).

Mass/charge (ESI) M+H: 489.0.

2-(4-(4-Acryloyl-1-piperazinyl)-7-chloro-1-phenyl-6-phthalazinyl)-3-fluorophenol (Example 31) was the second elutable regioisomer obtained from the reverse phase column.

1H NMR (400 MHz, CHLOROFORM-d), δ: 8.15 (1H, s), 8.12 (1H, s), 7.68-7.73 (2H, m), 7.53-7, 58 (3H, m), 7.30 (1H, br td, J=8.22, 6.65 Hz), 6.88 (1H, d, J=8.22 Hz), 6.78 (1H, t, J=8.61 Hz), 6.57 (1H, dd, J=16.82, 10.56 Hz), 6.28 (1H, dd, J=16.73, 1.66 Hz), 5.71 (1H, dd, J=10.56, 1.56 Hz), 3.78-3.89 (4H, m), 3.51-3.73 (4H, m).

Mass/charge (ESI) M+H: 489.1.

- 129 044387

Examples 32 and 33.

32) and

2-(1-(4-Acryloyl-1-piperazinyl)-7-chloro-4-methoxy-6-phthalazinyl)-3-fluorophenol (example 2-(4-(4-acryloyl-1-piperazinyl)-7- chloro-1-methoxy-6-phthalazinyl)-3-fluorophenol (example 33)

Examples 32 and 33 were prepared in a manner similar to examples 30 and 31, with the exception of step 3, which was modified as follows.

Step 3. tert-Butyl-4-(6,7-dichloro-4-methoxyphthalazin-1-yl)piperazine-1-carboxylate.

tert-Butyl-4-(4,6,7-trichlorophthalazin-1-yl)piperazine-1-carboxylate (intermediate M, 198 mg, 0.474 mmol) and sodium methoxide (25% solution in methanol, 2 ml, 8. 75 mmol) were mixed in a sealed vial. The reaction mixture was stirred at 60°C for 2 hours. The reaction mixture was quenched with saturated aqueous sodium bicarbonate (25 ml) and extracted with EtOAc (25 ml). The organic layer was separated, washed with brine (20 ml), dried over MgSO ₄ , filtered and concentrated in vacuo. Chromatographic purification of the residue (silica gel, 0 to 50% EtOAc in heptane) provided tert-butyl 4-(6,7-dichloro-4methoxyphthalazin-1-yl)piperazine-1-carboxylate.

1H NMR (400 MHz, CHLOROFORM-d), δ: 8.29 (1H, s), 8.08 (1H, s), 4.22 (3H, s), 3.68-3.73 (4H, m), 3.333.38 (4H, m), 1.51 (9H, s).

Mass/charge (ESI) M+H: 413.1.

From Step 6. First regioisomer to elute: 2-(1-(4-acryloyl-1-piperazinyl)-7-chloro-4methoxy-6-phthalazinyl)-3-fluorophenol (Example 32).

1H NMR (400 MHz, CHLOROFORM-d), δ: 8.23 (1H, s), 8.11 (1H, s), 7.32 (1H, td, J=8.31, 6.46 Hz) , 6.88 (1H, d, J=8.22 Hz), 6.77-6.83 (1H, m), 6.65 (1H, dd, J=16.82, 10.56 Hz), 6.37 (1H, dd, J=16.82, 1.76 Hz), 5.79 (1H, dd, J=10.47, 1.86 Hz), 4.18 (3H, s), 3 .79-4.05 (4H, m), 3.34-3.54 (4H, m).

Mass/charge (ESI) M+H: 443.1.

Second regioisomer eluted: 2-(4-(4-acryloyl-1-piperazinyl)-7-chloro-1-methoxy-6phthalazinyl)-3-fluorophenol (Example 33).

1H NMR (400 MHz, CHLOROFORM-d), δ: 8.32 (1H, s), 8.01 (1H, s), 7.32 (1H, td, J=8.27, 6.55 Hz) , 6.89 (1H, d, J=8.22 Hz), 6.77-6.83 (1H, m), 6.60 (1H, dd, J=17.02, 10.56 Hz), 6.30 (1H, dd, J=16.82, 1.76 Hz), 5.75 (1H, dd, J=10.56, 1.76 Hz), 4.22 (3H, s), 3 .67-3.98 (4H, m), 3.25-3.55 (4H, m).

Mass/charge (ESI) M+H: 443.1.

Example 34.

-(4-Acryloyl-1-piperazinyl)-4-benzyl-6,7-dichlorophthalazine

Example 34 was prepared in a manner similar to Examples 30 and 31, with the exception of step 6, which was omitted, and steps 2 and 3, which were modified as follows.

Steps 2 and 3. tert-Butyl 4-(4-benzyl-6,7-dichlorophthalazin-1-yl)piperazine-1-carboxylate.

Benzylzinc bromide (0.5 M in THF, 1.926 mL, 0.963 mmol) was added to a sealed vial containing 1,4,6,7-tetrachlorophthalazine (intermediate L, 258 mg, 0.963 mmol) and tetrakis(triphenylphosphine)palladium (111 mg, 0.096 mmol, Strem Chemicals Inc., Newburyport, MA, USA) under argon. The reaction mixture was stirred at room temperature. over 16 hours. 1-Vospiperazine (1.79 g, 9.63 mmol) was added and the reaction mixture was stirred at 60°C for 5 hours. The reaction mixture was quenched with saturated aqueous sodium bicarbonate (40 ml) and extracted with EtOAc (50 ml). The organic layer was separated, washed with brine (40 ml), dried over MgSO ₄ , filtered and concentrated in vacuo. Chromatographic purification of the residue (silica gel, 0 to 50% EtOAc in heptane) provided tert-butyl 4-(4-benzyl-6,7dichlorophthalazin-1-yl)piperazine-1-carboxylate.

1H NMR (400 MHz, CHLOROFORM-d) δ: 8.11 (1H, s) 8.10 (1H, s), 7.27-7.35 (4H, m), 7.20-7.25 ( 1Н, m)

- 130 044387

4.59 (2H, s) 3.69-3.74 (4H, m) 3.44-3.49 (4H, m) 1.52 (9H, s).

Mass/charge (ESI) M+H: 473.1.

From step 5. 1-(4-Acryloyl-1-piperazinyl)-4-benzyl-6,7-dichlorophthalazine.

1H NMR (400 MHz, CHLOROFORM-d), δppm 8.13 (1H, s), 8.12 (1H, s), 7.28-7.36 (4H, m), 7.20-7.26 (1H, m), 6.65 (1H, dd, J=16.82, 10.56 Hz), 6.37 (1H, dd, J=16.82, 1.57 Hz), 5.78 ( 1H, dd, J=10.56, 1.56 Hz), 4.61 (2H, s), 3.83-4.01 (4H, m), 3.48-3.62 (4H, m) .

Mass/charge (ESI) M+H: 427.1.

Examples 35 and 36.

2-(1-(4-Acryloyl-1-piperazinyl)-4-benzyl-7-chloro-6-phthalazinyl)-3-fluorophenol (Example 35) and 2-(4-(4-acryloyl-1-piperazinyl) -1-benzyl-7-chloro-6-phthalazinyl)-3-fluorophenol (example 36)

1-(4-(4-Benzyl-6,7-dichlorophthalazin-1-yl)piperazin-1-yl)prop-2-en-1-one. (Example 34, 35 mg, 0.082 mmol), 2-fluoro-6-hydroxyphenylboronic acid (12.77 mg, 0.082 mmol, Combi-Blocks Inc., San Diego, CA, USA), SPhos Pd G3 (7.09 mg, 8.19 µmol) and sodium carbonate (2 M aq, 0.123 ml, 0.246 mmol) were mixed in 1,2-dimethoxyethane (0.3 ml) in a sealed vial under argon. The reaction mixture was stirred at 60°C for 1 hour. The reaction mixture was quenched with saturated aqueous sodium bicarbonate (15 ml) and extracted with EtOAc (20 ml). The organic layer was separated, washed with brine (10 ml), dried over MgSO ₄ , filtered and concentrated in vacuo. Chromatographic purification of the residue (silica gel, 0 to 100% EtOAc in heptane) provided a mixture of two regioisomeric products. Preparative reverse phase chromatography (XBridge Prep C18 5 µm OBD, 150x30 mm; 20 to 90% (0.1% TFA in water) in (0.1% TFA in acetonitrile); flow = 30 ml/min.) provided obtaining partially separated regioisomeric products. Fractions containing regioisomers were neutralized with saturated aqueous sodium bicarbonate and extracted with DCM, and the organic extracts were concentrated in vacuo. 2-(1-(4-Acryloyl-1-piperazinyl)-4-benzyl-7-chloro-6phthalazinyl)-3-fluorophenol (Example 35) was the first regioisomer eluted during preparative reverse phase chromatography and contained approximately 36% of the second eluted regioisomer.

^1H NMR (400 MHz, CHLOROFORM-d), δ: 8.13 (1H, s), 8.11 (1H, s), 7.12-7.37 (6H, m), 6.91 (1H , d, J=8.22 Hz), 6.77 (1H, t, J=8.61 Hz), 6.64 (1H, dd, J=16.82, 10.56 Hz), 6.37 (1H, dd, J=16.82, 1.76 Hz), 5.79 (1H, dd, J=10.56, 1.96 Hz), 4.55 (2H, s), 3.34- 4.01 (8H, m).

Mass/charge (ESI) M+H: 503.1.

2-(4-(4-Acryloyl-1-piperazinyl)-1-benzyl-7-chloro-6-phthalazinyl)-3-fluorophenol (Example 36) was the second regioisomer to elute.

1H NMR (400 MHz, CHLOROFORM-d) δ: 8.12 (1H, s), 8.05 (1H, s), 7.26-7.36 (5H, m), 7.19-7.24 (1H, m), 6.93 (1H, d, J=8.41 Hz), 6.76 (1H, t, J=8.31 Hz), 6.58 (1H, dd, J=16, 82, 10.76 Hz), 6.28 (1H, dd, J=16.82, 1.76 Hz), 5.75 (1H, dd, J=10.56, 1.76 Hz), 4, 54 (2H, s), 3.32-3.93 (8H, m).

Mass/charge (ESI) M+H: 503.1.

Example 37.

1-(4-Acryloyl-1-piperazinyl)-4-benzyl-6-chloro-7-(5-methyl-1H-indazol-4-yl)phthalazine and 1-(4-acryloyl-1-piperazinyl)-4 -benzyl-7-chloro-6-(5-methyl-1H-indazol-4-yl)phthalazine

Example 37 was prepared similarly to Examples 35 and 36 using 5-methyl-1H-inαzol-4ylboronic acid (Combi-Blocks Inc., San Diego, CA, USA) instead of 2-fluoro-6-131 044387 hydroxyphenylboronic acid. In this embodiment, the two regioisomeric products were not separated.

1H NMR (400 MHz, CHLOROFORM-d) δ: 8.23 (0.6H, s), 8.22 (0.4H, s), 8.02 (0.4H, s), 8.00 (0 ,6Н, s), 7.19-7.57 (8Н, m), 6.68 (0.4Н, dd, J=16.82, 10.56 Hz), 6.60 (0.6Н, dd , J=16.82, 10.56 Hz), 6.38 (0.4H, dd, J=16.63, 1.76 Hz), 6.32 (0.6H, dd, J=16.82 , 1.76 Hz), 5.79 (0.4 N, dd, J=10.56, 1.76 Hz), 5.73 (0.6 N, dd, J=10.56, 1.76 Hz ), 4.67 (1.2H, s), 4.60 (0.8H, s), 3.74-4.06 (4H, m), 3.46-3.70 (4H, m), 2.21 (1.8H, s), 2.06 (1.2H, s).

Mass/charge (ESI) M+H: 523.

Example 38.

6-Chloro-1-(cyclopropylmethyl)-7-(2-fluoro-6-hydroxyphenyl)-4-(4-(2-propenoyl)-1-piperazinyl)pyrido[2,3-b]pyrimidine-2(1H )-oh

The starting material for example 38 was prepared using method 8, steps 1-4, using the reagents: 2,5,6-trichloronicotinic acid (step 1), aminomethylcyclopropane (step 2), 2-fluoro-6-hydroxyphenylboronic acid (step 4 , Combi-Blocks Inc., San Diego, California, USA) and sodium carbonate (step 4).

Step 1. 7-(2-((tert-Butyldiphenylsilyl)oxy)-6-fluorophenyl)-6-chloro-1-(cyclopropylmethyl)pyrido[2,3-d]pyrimidine-2,4(1H,3H)- dione

tert-Butylchlorodiphenylsilane (0.036 ml, 0.139 mmol) was added to a stirred mixture of 6-chloro1-(cyclopropylmethyl)-7-(2-fluoro-6-hydroxyphenyl)pyrido[2,3-d]pyrimidine-2,4(1H,3H )-dione (42 mg, 0.116 mmol) and triethylamine (0.065 ml, 0.464 mmol) in acetonitrile (0.5 ml). The reaction mixture was stirred at room temperature. for 1 hour. The reaction mixture was quenched with saturated aqueous NH ₄ Cl (25 ml) and extracted with EtOAc (30 ml). The organic layer was separated, washed with brine (25 ml), dried over MgSO ₄ , filtered and concentrated in vacuo to give crude 7-(2-((tert-butyldiphenylsilyl)oxy)-6-fluorophenyl)-6-chloro- 1-(cyclononylmethyl)nirido[2,3-d]nirimidine-2,4(1H,3H)-dione, which was used directly in the next step.

Mass/charge (ESI) M+H: 599.8.

Step 2. 4-(4-Acryloylpiperazin-1-yl)-7-(2-((tert-butyldiphenylsilyl)oxy)-6-fluorophenyl)-6chloro-1-(cyclopropylmethyl)πyrido[2,3-d]pyrimidine -2(1H)-one.

Phosphorus oxychloride (0.087 mL, 0.933 mmol) was added to a stirred mixture of crude 7-(2-((tert-butyldiphenylsilyl)oxy)-6-fluorophenyl)-6-chloro-1-(cyclopropylmethyl)πpyrido[2,3-d] pyrimidine-2,4(1H,3H)-dione (70 mg, 0.117 mmol), triethylamine (0.295 ml, 2.099 mmol) and 1H-benzo[d][1,2,3]triazole (167 mg, 1,400 mmol) in acetonitrile (2 ml). The reaction mixture was stirred at 80°C for 4 hours. The reaction mixture was concentrated under vacuum. The resulting residue was taken up in 1,2-dichloroethane (2 ml) and triethylamine (0.295 ml, 2.099 mmol) and 1-(piperazin-1yl)prop-2-en-1-one (32.7 mg, 0.233 mmol, eNovation) were added Chemicals LLC, Bridgewater, New Jersey, USA). The reaction mixture was stirred at room temperature. over 16 hours. Additional triethylamine (0.148 mL, 1.050 mmol) and 1-(piperazin-1-yl)prop-2-en-1-one (32.7 mg, 0.233 mmol, eNovation Chemicals LLC, Bridgewater, New Jersey, USA) and the reaction mixture was stirred at room temperature. for 1 hour before heating to 60°C and stirring for 4 hours. The reaction mixture was diluted with saturated aqueous NaHCO ₃ (40 ml) and extracted with DCM (50 ml). The organic layer was separated, dried over MgSO ₄ , filtered and concentrated in vacuo. The resulting residue was taken up again with 1,2-dichloroethane (2 ml) and triethylamine (0.295 ml, 2.099 mmol) and 1-(piperazin-1-yl)prop-2-en-1-one (32.7 mg, 0.233 mmol, eNovation Chemicals LLC, Bridgewater, NJ, USA). The reaction mixture was stirred at 60°C for 6 hours. The reaction mixture was diluted with saturated aqueous NaHCO ₃ (40 ml) and extracted with DCM (50 ml). The organic layer was separated, dried over MgSO ₄ , filtered and concentrated in vacuo. Chromatographic purification of the residue (silica gel, 0 to 100% (3:1 EtOAc/EtOH) in heptane) provided 4-(4-acryloylpiperazin-1-yl)-7-(2-((tert-butyldiphenylsilyl)oxy)- 6-fluorophenyl)-6-chloro-1(cyclopropylmethyl)pyrido[2,3^]pyrimidin-2(1H)-one, which was taken to the next stage without additional purification.

Mass/charge (ESI) M+H: 721.8.

Step 3. 4-(4-Acryloylpiperazin-1-yl)-6-chloro-1-(cyclopropylmethyl)-7-(2-fluoro-6-hydroxyphenyl)pyrido[2,3-d]pyrimidin-2(1H) -He.

Tetrabutylammonium fluoride (1.0 M solution in tetrahydrofuran, 0.025 ml, 0.025 mmol) was added to the stirred mixture of 4-(4-acryloylpiperazin-1-yl)-7-(2-((tert-butyldiphenylsilyl)oxy)-6-fluorophenyl )-6-chloro-1-(cyclopropylmethyl)pyrido[2,3-d]pyrimidin-2(1H)-one (6 mg, 8.31 µmol) in tetrahydrofuran (0.2 ml). The reaction mixture was stirred at room temperature. for 20 min before concentrating in vacuo. Chromatographic purification of the residue (silica gel, 0 to 100% (3:1 EtOAc/EtOH) in heptane) provided 6-chloro-1-(cyclopropylmethyl)-7-(2-fluoro-6-hydroxyphenyl)-4-(4 -(2propenoyl)-1-piperazinyl)pyrido[2,3-d]pyrimidin-2(1H)-one.

¹ H NMR (400 MHz, CHLOROFORM-d), δ: 8.04 (1H, s), 7.26-7.33 (1H, m), 6.82 (1H, d, J=8.29 Hz ), 6.71 (1H, t, J=8.91 Hz), 6.51 (1H, dd, J=16.79, 10.57 Hz), 6.30 (1H, dd, J=16, 79, 1.45 Hz), 5.72 (1H, dd, J=10.47, 1.55 Hz), 4.15 (2H, br d, J=6.43 Hz), 3.69-3 .90 (8H, m), 1.14-1.27 (4H, m), 0.73-0.88 (1H, m).

Mass/charge (ESI) M+H: 483.8.

Example 39.

6-Chloro-7-(2-fluoro-6-hydroxyphenyl)-1-(2-methyl-6-(2-propanyl)phenyl)-4-((2S)-2-methyl-4-(2propenoyl)- 1-piperazinyl)pyrido[2,3-d]pyrimidin-2(1H)-one

[1)(COCI) ₂ , THF, 65 °C

Intermediate P (1) POCI ₃₁ DIPEA, Pd(dppf)CI ₂ KOAc,

Step 1. 2,5,6-Trichloronicotinamide (intermediate P).

1,1'-Carbonyldiimidazole (40 g, 247 mmol) was added portionwise to 2,5,6-trichloronicotinic acid (50.7 g, 224 mmol, Combi-Blocks, San Diego, CA, USA) in THF (400 ml ), ensuring that gas evolution ceases between additions. The resulting mixture was stirred for 5 min and then degassed under vacuum and flushed with nitrogen (x2). The resulting mixture was heated to 50°C for 60 min, then diluted with toluene (100 ml) and concentrated to half volume. The resulting mixture was cooled to 0°C and ammonium hydroxide (60 ml, 437 mmol) was added slowly using a syringe. The reaction mixture was stirred for 10 min at room temperature, diluted with EtOAc (200 ml) and washed with water (3 x 100 ml). The organic layer was dried over anhydrous sodium sulfate and concentrated. The residue was suspended in 9:1 heptane/EtOAc (300 ml) and filtered. The filtered solids were collected and the remaining stock solution was partially evaporated to half volume, cooled to 0°C and filtered. Two portions of the filtered solids were combined to obtain 2,5,6-trichloronicotinamide.

Step 2. 2,5,6-Trichloro-N-((2-isopropyl-6-methylphenyl)carbamoyl)nicotinamide.

To a mixture of 2,5,6-trichloronicotinamide (intermediate P, 1.13 g, 5.0 mmol) in THF (30 mL) was added oxalyl chloride (2 M solution in DCM, 2.7 mL, 5.4 mmol). The resulting suspension was heated at 65°C for 40 minutes, then the heating was stopped and the reaction mixture was cooled to room temperature. Add 2-isopropyl-6-methylaniline (0.80 ml, 5.36 mmol, Enamine,

- 133 044387

Monmouth Junction, New Jersey, USA) and the reaction mixture was stirred at room temperature for 14 hours. The reaction mixture was concentrated and the residue was partitioned between EtOAc (50 ml) and saturated aqueous sodium bicarbonate (10 ml). The organic layer was washed with brine (10 ml), dried over anhydrous sodium sulfate and concentrated. The residue was suspended in 5:1 heptane/EtOAc (10 ml) and filtered. The filtered solids were collected to give 2,5,6-trichloro-K-((2-isopropyl-6-methylphenyl)carbamoyl)nicotinamide.

1H NMR (400 MHz, CHLOROFORM-d) δ: ppm 9.63 (s, 1H), 9.35 (br s, 1H), 8.25 (s, 1H), 7.19-7.26 (m , 2H), 7.13 (d, J=7.3 Hz, 1H), 3.14 (quin, J=6.9 Hz, 1H), 2.29 (s, 3H), 1.23 (d , J=6.8 Hz, 6H).

Mass/charge (ESI, +ve ion): 400.0 (M+H) ⁺ .

Step 3. 6,7-Dichloro-1-(2-isopropyl-6-methylphenyl)pyrido[2,3-d]pyrimidine-2,4(1H,3H)-dione.

To a mixture of 2,5,6-trichloro-K-((2-isopropyl-6-methylphenyl)carbamoyl)nicotinamide (1.45 g, 3.6 mmol) in THF (20 ml) was added KHMDS (1 M solution in THF , 7.5 ml, 7.5 mmol). After stirring for 30 min at room temperature, the reaction mixture was concentrated to 1/3 volume and quenched with saturated aqueous ammonium chloride (10 ml). The mixture was extracted with EtOAc (40 ml). The organic layer was washed with brine (10 ml), dried over anhydrous sodium sulfate and concentrated to give 6,7-dichloro-1-(2-isopropyl-6methylphenyl)pyrido[2,3-b]pyrimidine-2,4( 1H,3H)-dione.

This material was used in the next step without further purification.

Mass/charge (ESI, +ve ion): 364.0 (M+H) ⁺ .

Step 4. (S)-tert-Butyl-4-(6,7-dichloro-1-(2-isopropyl-6-methylphenyl)-2-oxo-1,2-dihydropyrido[2,3-d]pyrimidin- 4-yl)-3-methylpiperazine-1-carboxylate.

To a mixture of crude 6,7-dichloro-1-(2-isopropyl-6-methylphenyl)pyrido[2,3-d]pyrimidine2,4(1H,3H)-dione (3.6 mmol) in acetonitrile (10 ml) DIPEA (1.50 mL, 8.6 mmol) was added followed by phosphorus oxychloride (0.50 mL, 5.3 mmol). The resulting mixture was heated at 80°C for 1 hour, then cooled to room temperature and concentrated. The residue was dissolved in DMF (15 ml) and treated with DIPEA (1.50 ml, 8.6 mmol), then with (S)-4-N-Boc-2methylpiperazine (900 mg, 4.5 mmol, ArkPharm Inc ., Arlington Heights, Illinois, USA). The resulting solution was stirred at room temperature for 14 hours and then diluted with EtOAc (30 ml). The mixture was washed with water (10 ml) and brine (10 ml), and the organic layer was dried over anhydrous magnesium sulfate and concentrated. The residue was purified by silica gel chromatography (eluent: 10-50% 3:1 EtOAc-EtOH/heptane) to give (S)-tert-butyl-4-(6,7dichloro-1-(2-isopropyl-6-methylphenyl) -2-oxo-1,2-dihydropyrido[2,3-d]pyrimidin-4-yl)-3methylpiperazine-1-carboxylate.

1H NMR (400 MHz, MeOH-d4) δ: ppm 8.45 (s, 1H), 7.34-7.43 (m, 2H), 7.23 (d, J=7.3 Hz, 1H) , 4.97 (br s, 1H), 4.34 (br d, J=13.3 Hz, 1H), 4.15 (br d, J=12.0 Hz, 1H), 4.01 (br d, J=13.7 Hz, 1H), 3.80 (br s, 1H), 3.093.32 (m, 2H), 2.49-2.59 (m, 1H), 1.99 (d, J=3.7 Hz, 3H), 1.55 (s, 9H), 1.50 (dd, J=1.7, 6.6 Hz, 3H), 1.18 (dd, J=6.7 , 1.8 Hz, 3H), 1.09 (dd, J=6.8, 2.3 Hz, 3H).

Mass/charge (ESI, +ve ion): 546.1 (M+H) ⁺ .

Step 5. (3S)-tert-Butyl-4-(6-chloro-7-(2-fluoro-6-hydroxyphenyl)-1-(2-isopropyl-6-methylphenyl)-2oxo-1,2-dihydropyrido[ 2,3-d]pyrimidin-4-yl)-3-methylpiperazine-1-carboxylate.

(S)-tert-butyl-4-(6,7-dichloro-1-(2-isopropyl-6-methylphenyl)-2oxo-1,2-dihydropyrido[2,3-d]pyrimidine-4 was loaded into a round-bottom flask -yl)-3-methylpiperazine-1-carboxylate (8.3 g, 15.19 mmol), potassium acetate (7.50 g, 76 mmol) and [1,1'-bis-(diphenylphosphino)ferrocene]-dichloropalladium (P), complex with dichloromethane (0.495 g, 0.606 mmol). 1,4-dioxane (40 ml) and water (8 ml) were added and the mixture was heated to 90°C. Add (2-fluoro-6-hydroxyphenyl)potassium trifluoroborate (intermediate Q, 7.45 g, 34.2 mmol) and additional [1,1'-bis(diphenylphosphino)ferrocene]-dichloropalladium(II) complex with dichloromethane (0.176 g). The resulting mixture was stirred at room temperature for 2.5 hours, then cooled to room temperature, diluted with EtOAc (200 ml) and washed with water (1x) and brine (1x). The organic layer was dried over anhydrous sodium sulfate and concentrated. The residue was purified by silica gel chromatography (eluent: 5-40% 3:1 EtOAc-EtOH/heptane) to give (3S)-tert-butyl-4-(6-chloro-7-(2-fluoro-6-hydroxyphenyl) -1-(2-isopropyl-6-methylphenyl)-2-oxo-1,2-dihydropyrido[2,3-d]pyrimidin-4-yl)-3-methylpiperazine-1-carboxylate.

1H NMR (400 MHz, CHLOROFORM-d) δ: ppm 8.68 (br s, 1H), 8.14 (br s, 1H), 7.33-7.45 (m, 2H), 7.25 ( d, J=6.3 Hz, 2H), 6.64-6.73 (m, 2H), 3.91-5.15 (m, 4H), 3.67 (br s, 1H), 3, 32 (br s, 2H), 2.49-2.76 (m, 1H), 1.952.08 (m, 3H), 1.53 (s, 12H), 1.14-1.29 (m, 3H ), 0.95-1.07 (m, 3H).

¹⁹ F NMR (377 MHz, CHLOROFORM-d) δ: ppm -104.56 (br s, 1F).

Mass/charge (ESI, +ve ion): 622.1 (M+H) ⁺ .

Step 6. 6-Chloro-7-(2-fluoro-6-hydroxyphenyl)-1-(2-methyl-6-(2-propanyl)phenyl)-4-((2S)-2-methyl-4(2 -propenoyl)-1-piperazinyl)pyrido[2,3-d]pyrimidin-2(1H)-one.

To a solution of (3S)-tert-butyl-4-(6-chloro-7-(2-fluoro-6-hydroxyphenyl)-1-(2-isopropyl-6-methylphenyl)

- 134 044387

2-oxo-1,2-dihydropyrido[2,3-d]pyrimidin-4-yl)-3-methylpiperazine-1-carboxylate (680 mg, 1.1 mmol) in DCM (10 ml) was added trifluoroacetic acid (1 .5 ml, 19.6 mmol). The reaction mixture was stirred for 1.5 hours at room temperature and then concentrated. The residue was partitioned between EtOAc (40 ml) and saturated aqueous sodium bicarbonate (2x15 ml). The organic layer was washed with brine (5 ml), dried over anhydrous sodium sulfate and concentrated. The residue was dissolved in DCM (10 ml) and treated with DIPEA (0.5 ml, 2.9 mmol) then with acryloyl chloride (0.09 ml, 1.1 mmol). The reaction mixture was stirred for 10 min at room temperature, then diluted with EtOAc (30 ml) and washed with saturated aqueous sodium bicarbonate (10 ml) and brine (5 ml). The organic layer was dried over anhydrous sodium sulfate and concentrated. The residue was purified by silica gel chromatography (eluent: 10-60% 3:1 EtOAc-EtOH/heptane) to give 6-chloro-7-(2fluoro-6-hydroxyphenyl)-1-(2-methyl-6-(2- propanyl)phenyl)-4-((2S)-2-methyl-4-(2-propenoyl)-1 piperazinyl)pyrido[2,3-d]pyrimidin-2(1H)-one.

^1H NMR (400 MHz, DMSO- _d6 ) δ: ppm 10.07 (s, 1H), 8.40 (br s, 1H), 7.17-7.25 (m, 3H), 7.06 -7.13 (m, 1H), 6.79-6.91 (m, 1H), 6.69 (d, J=8.0 Hz, 1H), 6.65 (t, J=8.8 Hz, 1H), 6.20 (br d, J=16.8 Hz, 1H), 5.73-5.78 (m, 1H), 4.91 (br s, 1H), 4.21-4 .46 (m, 2H), 3.95-4.20 (m, 1H), 3.42-3.80 (m, 2H), 3.03-3.27 (m, 1H), 2.53 -2.63 (m, 1H), 1.85 (br s, 3H), 1.32 (br t, J=5.9 Hz, 3H), 1.05 (d, J=6.8 Hz, 3H), 0.91 (br d, J=6.6 Hz, 3H).

¹⁹ F NMR (377 MHz, DMSO-d ₆ ) δ: ppm -115.71 - -115.50 (m, 1F), -116.16 (br s, 1F). Mass/charge (ESI, +ve ion): 576.0 (M+H) ⁺ .

__7 KF, L-(+) -tartaric acid U <Qa(oh) ₂ ------------- Pj^bf ₃ -k~

OH MeCN/THF, b. t. OH

Intermediate Q (2-Fluoro-6-hydroxyphenyl)potassium trifluoroborate (intermediate Q).

A solution of potassium fluoride (44.7 g, 770 mmol) in water (75 ml) was added to a suspension of (2-fluoro-6hydroxyphenyl)boronic acid (30 g, 192 mmol, Combi-Blocks, San Diego, CA, USA) in acetonitrile (750 ml). The mixture was stirred for 2 minutes and then a solution of L-(+)-tartaric acid (72.2 g, 481 mmol) in THF (375 ml) was added over a period of 10 minutes. using a dropping funnel. The mixture was stirred vigorously using a mechanical stirrer for 1 hour, and then the resulting suspension was filtered and the filtered solids were washed with a small amount of THF. The solids were discarded and the filtrate was partially concentrated until the solids began to precipitate out of solution. The mixture was then cooled to -20°C and stirred for 16 hours. The reaction mixture was slowly heated and 2-propanol (20 ml) was added. The resulting suspension was filtered and the filtered solids were washed with 2-propanol. The filtrate was again partially concentrated until a suspension was formed and then cooled to -20°C and stirred for an additional 20 minutes. The resulting suspension was diluted with 2-propanol and filtered, and the filtered solids were washed with 2-propanol. Two batches of solids were combined to produce 2-fluoro-6hydroxyphenyl)potassium trifluoroborate (intermediate Q).

1H NMR (400 MHz, DMSO-d ₆ ) δ: ppm 8.07 (q, J=14.7 Hz, 1H), 6.93 (q, J=7.5 Hz, 1H), 6.30- 6.38 (m, 2H).

- 135 044387

Example 40.

6-Chloro-7-(2-fluorophenyl)-1-(4-methyl-2-(2-propanyl)-3-pyridinyl)-4-((2S)-2-methyl-4-(2-propenoyl) 1-piperazinyl)pyrido[2,3-d]pyrimidin-2(1H)-one _N Pd(dppf) ₂ CI ₂ .

„ I i-PrZnBr

Ti T THF, 60 °C

Step 1 Intermediate R (1)(COCI) ₂ , THF, 60 °C (2) TEA,

connection P

Step 1: 2-Isopropyl-4-methylpyridin-3-amine (intermediate R).

[1,1-Bis(diphenylphosphino )ferrocene]dichloropalladium(II), complex with dichloromethane (79 mg, 0.10 mmol). The resulting slurry was deoxygenated with argon for 2 min and then 2-propylzinc bromide (0.5 M solution in THF, 5.40 mL, 2.7 mmol, Sigma-Aldrich, St. Louis, MO, USA) was added. The resulting solution was heated at 60°C for 17 hours, then the heating was stopped and the reaction mixture was cooled to room temperature. The reaction mixture was quenched with water (10 ml) and 1 N. NaOH solution (20 ml) and then extracted with EtOAc (2x). The combined organic layers were dried over anhydrous sodium sulfate and concentrated. The residue was purified by silica gel chromatography (eluent: 0-15% MeOH/DCM) to give 2-isopropyl-4-methylpyridin-3-amine.

¹ H NMR (400 MHz, DMSO-d ₆ ) δ: ppm 7.66 (d, J=4.6 Hz, 1H), 6.78 (d, J=4.8 Hz, 1H), 4.72 (br s, 2H), 3.14-3.25 (m, 1H), 2.08 (s, 3H), 1.14 (d, J=6.8 Hz, 6H).

Mass/charge (ESI, +ve ion): 151.1 (M+H) ⁺ .

Step 2. 2,5,6-Trichloro-N-((2-isopropyl-4-methylpyridin-3-yl)carbamoyl)nicotinamide.

To a slurry of 2,5,6-trichloronicotinamide (intermediate P, 3.10 g, 13.8 mmol) in THF (46 ml) at -78° C., oxalyl chloride (2 M solution in DCM, 7.4 ml, 14.7 mmol) using a syringe. The resulting suspension was heated at 60°C for 3.5 hours, then the heating was stopped and the reaction mixture was cooled to -78°C. Triethylamine (6.0 mL, 42.6 mmol) was added followed by the addition of a solution of 2-isopropyl-4-methylpyridin-3-amine (intermediate R, 2.12 g, 14.1 mmol) via cannula. The resulting slurry was warmed to room temperature and stirred for 1 hour, then partitioned between water (120 ml) and EtOAc (175 ml). The organic layer was dried over anhydrous sodium sulfate and concentrated. The residue was suspended in heptane/EtOAc 9:1 and filtered. The filtered solids were collected to give 2,5,6-trichloro-N-((2isopropyl-4-methylpyridin-3-yl)carbamoyl)nicotinamide.

1H NMR (400 MHz, DMSO-d ₆ ) δ: ppm 11.31 (s, 1H), 9.54 (s, 1H), 8.66 (s, 1H), 8.34 (d, J=4 .8 Hz, 1H), 7.16 (d, J=5.0 Hz, 1H), 3.24-3.33 (m, 1H), 2.22 (s, 3H), 1.17 (d , J=6.6 Hz, 6H).

Mass/charge (ESI, +ve ion): 400.9 (M+H) ⁺ .

Step 3. 6,7-Dichloro-1-(2-isopropyl-4-methylpyridin-3-yl)pyrido[2,3-d]pyrimidine-2,4(1H,3H)dione.

To an ice-cooled solution of 2,5,6-trichloro-N-((2-isopropyl-4-methylpyridin-3-yl)carbamoyl)nicotinamide (4.71 g, 11.7 mmol) in THF (55 mL) was added slowly KHMDS (1 M solution in THF, 23.5 ml, 23.5 mmol) using a syringe. After 10 minutes, the ice bath was removed and the resulting solution

- 136 044387 was stirred for an additional 30 minutes at room temperature. The reaction mixture was quenched with saturated aqueous ammonium chloride (125 ml) and extracted with EtOAc (250 ml). The organic layer was washed with brine (1x), dried over anhydrous sodium sulfate and concentrated. The residue was purified by silica gel chromatography (eluent: 0-11% MeOH/DCM) to give 6,7-dichloro-1-(2-isopropyl-4-methylpyridin-3yl)pyrido[2,3-d]pyrimidine-2, 4(1H,3H)-dione.

^1H NMR (400 MHz, DMSO-d6) δ: ppm 12.27 (br s, 1H), 8.59 (s, 1H), 8.52 (d, J=5.0 Hz, 1H), 7 .28 (d, J=5.0 Hz, 1H), 2.82-2.92 (m, 1H), 2.04 (s, 3H), 1.08 (d, J=6.6 Hz, 3H), 1.01 (d, J=6.8 Hz, 3H).

Mass/charge (ESI, +ve ion): 365.0 (M+H) ⁺ .

Step 4. 4,6,7-Trichloro-1-(2-isopropyl-4-methylpyridin-3-yl)pyrido[2,3-d]pyrimidin-2(1H)-one.

To a suspension of 6,7-dichloro-1-(2-isopropyl-4-methylpyridin-3-yl)pyrido[2,3-d]pyrimidine-2,4(1H,3H)dione (2.52 g, 6. 9 mmol) in acetonitrile (45 ml) was added DIPEA (1.80 ml, 10.3 mmol) followed by slow addition of phosphorus oxychloride (1.58 ml, 10.3 mmol) using a syringe. The resulting mixture was heated at 80°C for 1.75 hours and then cooled to room temperature and concentrated to give 4,6,7-trichloro-1-(2-isopropyl-4-methylpyridin-3-yl)pyrido[2 ,3-d]pyrimidin2(1H)-one. This material was used in the next step without further purification.

Step 5. (S)-tert-Butyl-4-(6,7-dichloro-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydropyrido[2,3- d]pyrimidin-4-yl)-3-methylpiperazine-1-carboxylate.

To a solution of 4,6,7-trichloro-1-(2-isopropyl-4-methylpyridin-3-yl)pyrido[2,3-d]pyrimidin-2(1H)-one (2.64 g, 6.9 mmol) in THF (40 ml) was added DIPEA (3.61 ml, 20.7 mmol) followed by (S)-4-N-Boc-2-methylpiperazine (2.07 g, 10.3 mmol, Combi- Blocks, Inc., San Diego, California, USA). The resulting solution was stirred at room temperature for 1.5 hours, and then ice water (60 ml) was added. The mixture was stirred for an additional 5 minutes, then extracted with EtOAc (3x). The combined organic layers were dried over anhydrous sodium sulfate and concentrated. The residue was purified by silica gel chromatography (eluent: 0-11% MeOH/DCM) to give (S)-tert-butyl-4-(6,7-dichloro-1-(2-isopropyl-4-methylpyridin-3-yl )-2-oxo1,2-dihydropyrido[2,3-d]pyrimidin-4-yl)-3-methylpiperazine-1-carboxylate.

1H NMR (400 MHz, DMSO-d ₆ ) δ: ppm 8.46 (dd, J=11.6, 5.2 Hz, 2H), 7.25 (d, J=4.8 Hz, 1H), 4.79-4.93 (m, 1H), 4.10-4.24 (m, 1H), 3.87-4.05 (m, 1H), 3.77-3.87 (m, 1H ), 3.62-3.76 (m, 1H), 2.99-3.25 (m, 2H), 2.55-2.69 (m, 1H), 1.94 (d, J=2 .5 Hz, 3H), 1.45 (s, 9H), 1.32 (br t, J=5.7 Hz, 3H), 1.06 (d, J=6.8 Hz, 3H), 1 .00 (d, J=6.6 Hz, 3H).

Mass/charge (ESI, +ve ion): 547.2 (M+H) ⁺ .

Step 6. (S)-tert-Butyl-4-(6-chloro-7-(2-fluorophenyl)-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo1,2-dihydropyrido[ 2,3-d]pyrimidin-4-yl)-3-methylpiperazine-1-carboxylate.

To a solution of (S)-tert-butyl-4-(6,7-dichloro-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydropyrido[2,3-d ]pyrimidin-4-yl)-3-methylpiperazine-1-carboxylate (1.02 g, 1.8 mmol) in 1,4-dioxane (17 ml) was added potassium acetate (914 mg, 9.3 mmol) and ( 2-fluorophenyl)boronic acid (313 mg, 2.2 mmol, Sigma-Aldrich, St. Louis, MO, USA). The mixture was bubbled with argon and then [1,1'-bis-(diphenylphosphino)ferrocene]dichloropalladium(II) complexed with dichloromethane (76 mg, 0.093 mmol) was added. The mixture was again bubbled with argon and heated at 90°C. After 30 s, three drops of water were added to the reaction mixture. Heating was continued at 90°C for 40 minutes and then the reaction mixture was allowed to cool to room temperature. Water (50 ml) and brine (4 ml) were added and the resulting mixture was extracted with EtOAc (2x). The combined organic layers were washed with brine (1x), dried over anhydrous sodium sulfate and concentrated. The residue was purified by silica gel chromatography (eluent: 0-9% MeOH/DCM) to give (S)-tert-butyl-4-(6-chloro-7-(2-fluorophenyl)-1-(2-isopropyl-4 -methylpyridin-3yl)-2-oxo-1,2-dihydropyrido[2,3-d]pyrimidin-4-yl)-3-methylpiperazine-1-carboxylate.

1H NMR (400 MHz, DMSO-d6) δ: ppm 8.43 (d, J=2.5 Hz, 1H), 8.39 (d, J=4.8 Hz, 1H), 7.47-7 .55 (m, 1H), 7.16-7.33 (m, 4H), 4.86-4.97 (m, 1H), 4.21-4.30 (m, 1H), 3, 90-4.06 (m, 2H), 3.80-3.89 (m, 1H), 3.67-3.78 (m, 1H), 3.04-3.16 (m, 1H), 2.65-2.75 (m, 1H), 1.93 (s, 3H), 1.48 (s, 9H), 1.36 (br d, J=6.6 Hz, 3H), 1, 06 (d, J=6.6 Hz, 3H), 0.94 (dd, J=6.6, 2.1 Hz, 3H).

Mass/charge (ESI, +ve ion): 607.0 (M+H) ⁺ .

Step 7. 6-Chloro-7-(2-fluorophenyl)-1-(4-methyl-2-(2-propanyl)-3-pyridinyl)-4-((2S)-2-methyl-4-(2propenoyl )-1-piperazinyl)pyrido[2,3-d]pyrimidin-2(1H)-one.

Trifluoroacetic acid (4.0 mL, 53.9 mmol) was added to a solution of (S)-tert-butyl-4-(6-chloro-7-(2fluorophenyl)-1-(2-isopropyl-4-methylpyridin-3- yl)-2-oxo-1,2-dihydropyrido[2,3-d]pyrimidin-4-yl)-3methylpiperazine-1-carboxylate (935 mg, 1.54 mmol) in DCM (20 ml). The resulting solution was stirred at room temperature for 1.5 hours and then concentrated. The residue was dissolved in DCM (12 ml), cooled to 0°C and treated with DIPEA (0.807 ml, 4.62 mmol), then with acryloyl chloride (0.131 ml, 1.62 mmol; added dropwise via syringe). The resulting solution was stirred at 0°C for 35 min, then quenched with saturated aqueous sodium bicarbonate (35 ml) and extracted with DCM (2x). The combined organic layers were dried over anhydrous sodium sulfate and concentrated. The residue was purified by silica gel chromatography (eluent: 0-11% MeOH/DCM) to give 6-chloro-7-(2-fluorophenyl)-1-(4-methyl-2-(2-propanyl)3-pyridinyl)- 4-((2S)-2-methyl-4-(2-propenoyl)-1-piperazinyl)pyrido[2,3-d]pyrimidin-2(1H)-one.

¹ H NMR (400 MHz, DMSO-d6) δ: ppm 8.46 (br d, J=4.6 Hz, 1H), 8.39 (d, J=4.8 Hz, 1H), 7.47 -7.55 (m, 1H), 7.16-7.34 (m, 4H), 6.78-6.94 (m, 1H), 6.15-6.26 (m, 1H), 5 .73-5.80 (m, 1H), 4.95 (br s, 1H), 4.36-4.45 (m, 0.5 H), 4.24-4.36 (m, 1. 5 H), 4.11-4.21 (m, 0.5 H), 3.98-4.08 (m, 0.5 H), 3.71-3.85 (m, 1H), 3 ,60-3.69 (m, 0.5 H), 3.41-3.53 (m, 0.5 H), 3.06-3.27 (m, 1H), 2.65-2, 75 (m, 1H), 1.94 (d, J=1.7 Hz, 3H), 1.34 (d, J=6.2 Hz, 3H), 1.07 (d, J=6.6 Hz, 3H), 0.94 (dd, J=6.5, 0.9 Hz, 3H).

Mass/charge (ESI, +ve ion): 560.9 (M+H)+.

Example 41.

6-Fluoro-7-(2-fluoro-6-hydroxyphenyl)-1-(4-methyl-2-(2-propanyl)-3-pyridinyl)-4-((2S)-2-methyl-4-( 2propenoyl)-1-piperazinyl)pyrido[2,3-d]pyrimidin-2(1H)-one (1TSOS1) ₂ , THF, 75 C

Step 1: 2,6-Dichloro-5-fluoronicotinamide (intermediate S).

To a mixture of 2,6-dichloro-5-fluoro-nicotinic acid (4.0 g, 19.1 mmol, AstaTech Inc., Bristol, PA) in dichloromethane (48 ml) was added oxalyl chloride (2 M solution in DCM, 11, 9 ml, 23.8 mmol), followed by the addition of a catalytic amount of DMF (0.05 ml). The reaction mixture was stirred at room temperature overnight and then concentrated. The residue was dissolved in 1,4-dioxane (48 ml) and cooled to 0°C. Ammonium hydroxide solution (28.0-30% NH ₃ base, 3.6 mL, 28.6 mmol) was added slowly using a syringe. The resulting mixture was stirred at 0°C for 30 minutes and then concentrated. The residue was diluted with 1:1 EtOAc/heptane and shaken for 5 minutes, then filtered. The filtered solids were discarded and the remaining stock solution was partially concentrated to half volume and filtered. The filtered solids were washed with heptane and dried in a reduced pressure oven (45°C) overnight to give 2,6-dichloro-5-fluoronicotinamide.

^1H NMR (400 MHz, DMSO-d6) δ: ppm 8.23 (d, J=7.9 Hz, 1H), 8.09 (br s, 1H), 7.93 (br s, 1H).

Mass/charge (ESI, +ve ion): 210.9 (M+H)+.

Step 2. 2,6-Dichloro-5-fluoro-N-((2-isopropyl-4-methylpyridin-3-yl)carbamoyl)nicotinamide.

To an ice-cooled slurry of 2,6-dichloro-5-fluoronicotinamide (intermediate S, 5.0 g, 23.9 mmol) in THF (20 mL) was added slowly oxalyl chloride (2 M solution in DCM, 14.4 mL, 28 .8 mmol) using a syringe. The resulting mixture was heated at 75°C for 1 hour, then the heating was stopped and the reaction mixture was concentrated to half the volume. After cooling to 0°C, THF (20 ml) was added followed by dropwise addition of a solution of 2-isopropyl-4-methylpyridin-3-amine (intermediate R, 3.59 g, 23.92 mmol) in THF (10 ml) using a cannula. The resulting mixture was stirred at 0°C for 1 hour and then quenched using a 1:1 mixture of saline and saturated aqueous ammonium chloride. The mixture was extracted with EtOAc (3x) and the combined organic layers were dried over anhydrous sodium sulfate and concentrated to give 2,6-dichloro-5-fluoro-N-((2-isopropyl-4-methylpyridin-3-yl)carbamoyl) nicotinamide. This material was used in the next step without further purification.

Mass/charge (ESI, +ve ion): 385.1 (M+H)+.

- 138 044387

Step 3. 7-Chloro-6-fluoro-1-(2-isopropyl-4-methylpyridin-3-yl)pyrido[2,3-d]pyrimidine2,4(1H,3H)-dione.

To an ice-cooled solution of 2,6-dichloro-5-fluoro-K-((2-isopropyl-4-methylpyridin-3yl)carbamoyl)nicotinamide (9.2 g, 24.0 mmol) in THF (40 mL) was added slowly KHMDS (1 M solution in THF, 50.2 ml, 50.2 mmol) using a syringe. The ice bath was removed and the resulting mixture was stirred for 40 minutes at room temperature. The reaction mixture was quenched with saturated aqueous ammonium chloride and extracted with EtOAc (3x). The combined organic layers were dried over anhydrous sodium sulfate and concentrated. The residue was purified by silica gel chromatography (eluent: 0-50% 3:1 EtOAc-EtOH/heptane) to give 7-chloro-6-fluoro-1-(2-isopropyl-4-methylpyridin-3-yl)pyrido[2 ,3-d]pyrimidine-2,4(1H,3H)-dione.

¹ H NMR (400 MHz, DMSO-d ₆ ) δ: ppm 12.27 (br s, 1H), 8.48-8.55 (m, 2H), 7.29 (d, J=4.8 Hz , 1H), 2.87 (quin, J=6.6 Hz, 1H), 1.99-2.06 (m, 3H), 1.09 (d, J=6.6 Hz, 3H), 1 .01 (d, J=6.6 Hz, 3H).

¹⁹ F NMR (376 MHz, DMSO-d ₆ ) δ: -126.90 (s, 1F).

Mass/charge (ESI, +ve ion): 349.1 (M+H) ⁺ .

Step 4. 4,7-Dichloro-6-fluoro-1-(2-isopropyl-4-methylpyridin-3-yl)pyrido[2,3-d]pyrimidin-2(1H)-one.

To a solution of 7-chloro-6-fluoro-1-(2-isopropyl-4-methylpyridin-3-yl)pyrido[2,3-d]pyrimidine2,4(1H,3H)-dione (4.7 g, 13 .5 mmol) and DIPEA (3.5 mL, 20.2 mmol) in acetonitrile (20 mL), phosphorus oxychloride (1.63 mL, 17.5 mmol) was added dropwise using a syringe. The resulting mixture was heated at 80°C for 1 hour and then cooled to room temperature and concentrated to give 4,7-dichloro-6-fluoro-1-(2-isopropyl-4-methylpyridin-3-yl)pyrido[2 ,3-d]pyrimidin-2(1H)-one. This material was used in the next step without further purification.

Mass/charge (ESI, +ve ion): 367.1 (M+H) ⁺ .

Step 5. (S)-tert-Butyl-4-(7-chloro-6-fluoro-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2dihydropyrido[2,3- d]pyrimidin-4-yl)-3-methylpiperazine-1-carboxylate.

To an ice-cooled solution of 4,7-dichloro-6-fluoro-1-(2-isopropyl-4-methylpyridin-3yl)pyrido[2,3-d]pyrimidin-2(1H)-one (13.5 mmol) in acetonitrile (20 ml) was added DIPEA (7.1 ml, 40.3 mmol) followed by (S)-4-N-Boc-2-methylpiperazine (3.23 g, 16.1 mmol, Combi-Blocks, Inc ., San Diego, California, USA). The resulting mixture was warmed to room temperature and stirred for 1 hour, then diluted with cold saturated aqueous sodium bicarbonate (200 ml) and EtOAc (300 ml). The mixture was stirred for an additional 5 minutes, the layers were separated and the aqueous layer was extracted with additional EtOAc (1x). The combined organic layers were dried over anhydrous sodium sulfate and concentrated. The residue was purified by silica gel chromatography (eluent: 0-50% EtOAc/heptane) to give (S)-tert-butyl-4-(7-chloro-6-fluoro-1-(2-isopropyl-4-methylpyridinium-3 -yl)-2-oxo-1,2-dihydropyrido[2,3-d]pyrimidin-4-yl)-3-methylpiperazine-1-carboxylate.

Mass/charge (ESI, +ve ion): 531.2 (M+H) ⁺ .

Step 6. (3S)-tert-Butyl-4-(6-fluoro-7-(2-fluoro-6-hydroxyphenyl)-1-(2-isopropyl-4-methylpyridin3-yl)-2-oxo-1, 2-dihydropyrido[2,3-d]pyrimidin-4-yl)-3-methylpiperazine-1-carboxylate.

A mixture of (S)-tert-butyl-4-(7-chloro-6-fluoro-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydropyrido[2,3- d]pyrimidin-4-yl)-3-methylpiperazin-1-carboxylate (4.3 g, 8.1 mmol), potassium trifluoro(2-fluoro-6hydroxyphenyl)borate (intermediate Q, 2.9 g, 10, 5 mmol), potassium acetate (3.2 g, 32.4 mmol) and [1,1'-bis-(diphenylphosphino)ferrocene]dichloropalladium(P), complexed with dichloromethane (661 mg, 0.81 mmol) in 1 ,4-dioxane (80 ml) was degassed with nitrogen for 1 min. Deoxygenated water (14 ml) was added and the resulting mixture was heated at 90°C for 1 hour. Allow the reaction mixture to cool to room temperature, quench it with semi-saturated aqueous sodium bicarbonate and extract with EtOAc (2x) and DCM (1x) . The combined organic layers were dried over anhydrous sodium sulfate and concentrated. The residue was purified by silica gel chromatography (eluent: 0-60% 3:1 EtOAc-EtOH/heptane) to give (3S)-tert-butyl-4-(6-fluoro-7-(2-fluoro-6-hydroxyphenyl) -1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo1,2-dihydropyrido[2,3-d]pyrimidin-4-yl)-3-methylpiperazin-1-carboxylate.

¹ H NMR (400 MHz, DMSO-d ₆ ) δ: ppm 10.19 (br s, 1H), 8.38 (d, J=5.0 Hz, 1H), 8.26 (dd, J=12 ,5, 9.2 Hz, 1H), 7.23-7.28 (m, 1H), 7.18 (d, J=5.0 Hz, 1H), 6.72 (d, J=8, 0 Hz, 1H), 6.68 (t, J=8.9 Hz, 1H), 4.77-4.98 (m, 1H), 4.24 (br t, J=14.2 Hz, 1H ), 3.93-4.08 (m, 1H), 3.84 (br d, J=12.9 Hz, 1H), 3.52-3.75 (m, 1H), 3.07-3 .28 (m, 1H), 2.62-2.74 (m, 1H), 1.86-1.93 (m, 3H), 1.43-1.48 (m, 9H), 1.35 (dd, J=10.8, 6.8 Hz, 3H), 1.26-1.32 (m, 1H), 1.07 (dd, J=6.6, 1.7 Hz, 3H), 0.93 (dd, J=6.6, 2.1 Hz, 3H).

¹⁹ F NMR (376 MHz, DMSO-d ₆ ) δ: -115.65 (s, 1F), -128.62 (s, 1F).

Mass/charge (ESI, +ve ion): 607.3 (M+H) ⁺ .

Step 7. 6-Fluoro-7-(2-fluoro-6-hydroxyphenyl)-1-(4-methyl-2-(2-propanyl)-3-pyridinyl)-4-((2S)-2methyl-4- (2-propenoyl)-1-piperazinyl)pyrido[2,3-d]pyrimidin-2(1H)-one.

Trifluoroacetic acid (25 mL, 324 mmol) was added to a solution of (3S)-tert-butyl-4-(6-fluoro-7-(2fluoro-6-hydroxyphenyl)-1-(2-isopropyl-4-methylpyridinium-3- yl)-2-oxo-1,2-dihydropyrido[2,3-d]- 139 044387 pyrimidin-4-yl)-3-methylpiperazine-1-carboxylate (6.3 g, 10.4 mmol) in DCM ( 30 ml). The resulting mixture was stirred at room temperature for 1 hour and then concentrated. The residue was dissolved in DCM (30 ml), cooled to 0°C and treated sequentially with DIPEA (7.3 ml, 41.7 mmol) and a solution of acryloyl chloride (0.849 ml, 10.4 mmol) in DCM (3 ml, added drop by drop using a syringe). The reaction mixture was stirred at 0°C for 10 min, then quenched with semi-saturated aqueous sodium bicarbonate and extracted with DCM (2x). The combined organic layers were dried over anhydrous sodium sulfate and concentrated. The residue was purified by silica gel chromatography (eluent: 0-100% 3:1 EtOAc-EtOH/heptane) to give 6-fluoro-7-(2-fluoro-6-hydroxyphenyl)-1-(4-methyl-2-( 2-propanyl)-3-pyridinyl)-4-((2S)-2-methyl-4-(2propenoyl)-1-piperazinyl)pyrido[2,3-d]pyrimidin-2(1H)-one.

1H NMR (400 MHz, DMSO-d ₆ ) δ: ppm 10.20 (s, 1H), 8.39 (d, J=4.8 Hz, 1H), 8.24-8.34 (m, 1H ), 7.23-7.32 (m, 1H), 7.19 (d, J=5.0 Hz, 1H), 6.87 (td, J=16.3, 11.0 Hz, 1H) , 6.74 (d, J=8.6 Hz, 1H), 6.69 (t, J=8.6 Hz, 1H), 6.21 (br d, J=16.2 Hz, 1H), 5.74-5.80 (m, 1H), 4.91 (br s, 1H), 4.23-4.45 (m, 2H), 3.97-4.21 (m, 1H), 3 .44-3.79 (m, 2H), 3.11-3.3 (m, 1H), 2.67-2.77 (m, 1H), 1.91 (s, 3H), 1.35 (d, J=6.8 Hz, 3H), 1.08 (d, J=6.6 Hz, 3H), 0.94 (d, J=6.8 Hz, 3H).

¹⁹ F NMR (376 MHz, DMSO-d6) δ: ppm -115.64 (s, 1F), -128.63 (s, 1F).

Mass/charge (ESI, +ve ion): 561.2 (m+H) ⁺ .

Example 42.

1-(2-Cyclopropyl-4-methyl-3-pyridinyl)-6-fluoro-7-(2-fluoro-6-hydroxyphenyl)-4-((2S)-2-methyl-4-(2propenoyl)-1 -piperazinyl)pyrido [2,3 -d] pyrimidine-2 (1 H)-oh _Νμ Pd(dppf) ₂ CI _{2 Br} ^C ' ^PrZnBr 'l TN^ ¹ THF, 70 °C

Stage 1 Intermediate T

(1) (COCI) _2l THF, 65 °C (2)

Pd(dppf) ₂ CI ₂ , KOAc,

> _c A (1)TFA, DCM, K. T. >

< ) / \ (2) DIPEA, / ) _F F / ^N \ / \ / \ acryloyl chloride )— n /=( )=\ nA ^dcm '°° ^c HA

Step 1. 2-Cyclopropyl-4-methylpyridin-3-amine (intermediate T).

To a suspension of 3-amino-2-bromo-4-picoline (4.0 g, 21.4 mmol, Combi-Blocks, San Diego, CA, USA) and the dichloro[1,1'-bis(diphenylphosphino) adduct ferrocene]palladium(II)dichloride with dichloromethane (1.78 g, 2.1 mmol) in THF (100 ml), cyclopropylzinc bromide (0.5 M solution in THF, 68.4 ml, 34.2 mmol, Sigma) was slowly added -Aldrich, St. Louis, MO, USA) using a dropping funnel. The resulting mixture was heated at 70°C for 6 hours, then the heating was stopped and the reaction mixture was cooled to room temperature. The reaction mixture was quenched with 5N. NaOH solution and extracted with EtOAc (1x). The organic layer was dried over anhydrous magnesium sulfate and concentrated. The residue was purified by silica gel chromatography (eluent: 0-50% 3:1 EtOAc-EtOH/heptane) to give 2-cyclopropyl-4-methylpyridin-3-amine.

1H NMR (400 MHz, CHLOROFORM-d) δ: ppm 7.84 (d, J=4.8 Hz, 1H), 6.82 (d, J=4.8 Hz, 1H), 3.82 (br s, 2H), 2.17 (s, 3H), 1.85 (quin, J=6.7 Hz, 1H), 0.92-0.99 (m, 4H).

Mass/charge (ESI, +ve ion): 149.1 (M+H) ⁺ .

Step 2. 2,6-Dichloro-X-((2-cyclopropyl-4-methylpyridin-3-yl)carbamoyl)-5-fluoronicotinamide.

To a solution of 2,6-dichloro-5-fluoronicotinamide (intermediate S, 3.0 g, 14.4 mmol) in THF (30 mL) was added slowly oxalyl chloride (2 M solution in DCM, 10.7 mL, 21.5 mmol) using a syringe. The resulting mixture was heated at 65°C for 2 hours, then the heating was stopped and the reaction

- 140 044387 the mixture was concentrated. The residue was dissolved in THF (30 ml) and added by cannula to a solution of 2-cyclopropyl-4-methylpyridin-3-amine (intermediate T, 2.1 g, 14.4 mmol) in THF (105 ml). The resulting mixture was stirred at room temperature for 3 hours, then quenched with water and extracted with EtOAc (3x). The combined organic layers were dried over anhydrous sodium sulfate and concentrated. The residue was purified by silica gel chromatography (eluent: 0-50% EtOAc/heptane) to give 2,6-dichloro-K-((2-cyclopropyl-4-methylpyridin3-yl)carbamoyl)-5-fluoronicotinamide.

¹ H NMR (400 MHz, CHLOROFORM-d) δ: ppm 9.85 (s, 1H), 9.68 (s, 1H), 8.29 (d, J=4.8 Hz, 1H), 7, 97 (d, J=7.0 Hz, 1H), 6.99 (d, J=4.8 Hz, 1H), 2.29 (s, 3H), 2.08-2.16 (m, 1H ), 1.08-1.13 (m, 2H), 0.95-1.02 (m, 2H).

Mass/charge (ESI, +ve ion): 383.0 (M+H) ⁺ .

Step 3. 7-Chloro-1-(2-cyclopropyl-4-methylpyridin-3-yl)-6-fluoropyrido[2,3-d]pyrimidine2,4(1H,3H)-dione.

To an ice-cooled solution of 2,6-dichloro-K-((2-cyclopropyl-4-methylpyridin-3-yl)carbamoyl)5-fluoronicotinamide (3.35 g, 8.7 mmol) in THF (30 mL) was added slowly KHMDS (1 M solution in THF, 17.5 ml, 17.5 mmol) using a syringe. The ice bath was removed and the resulting mixture was stirred for 14 hours at room temperature. The reaction mixture was quenched with saturated aqueous ammonium chloride and extracted with EtOAc (3x). The combined organic layers were dried over anhydrous sodium sulfate and concentrated to give 7-chloro-1-(2cyclopropyl-4-methylpyridin-3-yl)-6-fluoropyrido[2,3-d]pyrimidine-2,4(1H,3H) -dione. This material was used without further purification in the next step.

Mass/charge (ESI, +ve ion): 347.0 (M+H) ⁺ .

Step 4. 4,7-Dichloro-1-(2-cyclopropyl-4-methylpyridin-3-yl)-6-fluoropyrido[2,3-d]pyrimidin2(1H)-one.

To a solution of 7-chloro-1-(2-cyclopropyl-4-methylpyridin-3-yl)-6-fluoropyrido[2,3-d]pyrimidine2,4(1H,3H)-dione (8.7 mmol) and DIPEA (2.32 mL, 13.1 mmol) in acetonitrile (25 mL), phosphorus oxychloride (1.22 mL, 13.1 mmol) was added dropwise using a syringe. The resulting mixture was heated at 80°C for 3 hours and then cooled to room temperature and concentrated to give 4,7-dichloro1-(2-cyclopropyl-4-methylpyridin-3-yl)-6-fluoropyrido[2,3- d]pyrimidin-2(1H)-one. This material was used in the next step without further purification.

Mass/charge (ESI, +ve ion): 365.0 (M+H) ⁺ .

Step 5. (S)-tert-Butyl-4-(7-chloro-1-(2-cyclopropyl-4-methylpyridin-3-yl)-6-fluoro-2-oxo-1,2dihydropyrido[2,3- d]pyrimidin-4-yl)-3-methylpiperazine-1-carboxylate.

To an ice-cooled solution of 4,7-dichloro-1-(2-cyclopropyl-4-methylpyridin-3-yl)-6fluoropyrido[2,3-d]pyrimidin-2(1H)-one (8.7 mmol) in DCM (25 mL) DIPEA (7.74 mL, 43.7 mmol) was added followed by (S)-4-N-Boc-2-methylpiperazine (1.75 g, 8.7 mmol, Combi-Blocks, Inc. , San Diego, California, USA). The resulting mixture was stirred at 0°C for 2 hours, then quenched with water and extracted with EtOAc (3x). The combined organic layers were dried over anhydrous sodium sulfate and concentrated. The residue was purified by silica gel chromatography (eluent: 0-60% EtOAc/heptane) to give (S)-tert-butyl-4-(7chloro-1-(2-cyclopropyl-4-methylpyridin-3-yl)-6- fluoro-2-oxo-1,2-dihydropyrido[2,3-d]pyrimidin-4-yl)3-methylpiperazine-1-carboxylate.

1H NMR (400 MHz, DMSO-d ₆ ) δ: ppm 8.31-8.39 (m, 2H), 7.18 (d, J=4.8 Hz, 1H), 4.82 (br s, 1H), 4.104.21 (m, 1H), 3.96 (br s, 1H), 3.82 (br d, J=13.3 Hz, 1H), 3.62-3.72 (m, 1H ), 3.19-3.52 (m, 2H), 1.94 (s, 3H), 1.62 (dq, J=8.2, 4.0 Hz, 1H), 1.45 (s, 9H), 1.32 (dd, J=6.5, 3.6 Hz, 3H), 0.87-0.98 (m, 1H), 0.69-0.84 (m, 2H), 0 .57-0.68 (m, 1H).

Mass/charge (ESI, +ve ion): 529.0 (M+H) ⁺ .

Step 6. (3S)-tert-Butyl-4-(1-(2-cyclopropyl-4-methylpyridin-3-yl)-6-fluoro-7-(2-fluoro-6hydroxyphenyl)-2-oxo-1, 2-dihydropyrido[2,3-d]pyrimidin-4-yl)-3-methylpiperazine-1-carboxylate.

Mixture of (S)-tert-butyl-4-(7-chloro-1-(2-cyclopropyl-4-methylpyridin-3-yl)-6-fluoro-2-oxo-1,2dihydropyrido[2,3-d] pyrimidin-4-yl)-3-methylpiperazine-1-carboxylate (987 mg, 1.87 mmol), (2-fluoro6-hydroxyphenyl)boronic acid (524 mg, 3.36 mmol, Combi-Blocks, San Diego, California, USA), potassium acetate (916 mg, 9.33 mmol) and [1,1'-bis-(diphenylphosphino)ferrocene]dichloropalladium(P), complexed with dichloromethane (152 mg, 0.19 mmol) in 1, 4-dioxane (10 ml) was bubbled with argon and heated at 80°C. After 2 minutes, three drops of water were added to the reaction mixture and the temperature was increased to 90°C. Heating was continued at 90°C for 1 hour and then the reaction mixture was allowed to cool to room temperature. Water was added and the resulting mixture was extracted with EtOAc (3x). The combined organic layers were washed with brine (1x), dried over anhydrous sodium sulfate and concentrated. The residue was purified by silica gel chromatography (eluent: 0-65% EtOAc/heptane) to give (3S)-tert-butyl-4-(1-(2cyclopropyl-4-methylpyridin-3-yl)-6-fluoro-7- (2-fluoro-6-hydroxyphenyl)-2-oxo-1,2-dihydropyrido[2,3d]pyrimidin-4-yl)-3-methylpiperazine-1-carboxylate.

- 141 044387

L NMR (400 MHz, DMSO-d ₆ ) δ: ppm 10.22 (s, 1H), 8.19-8.31 (m, 2H), 7.23-7.32 (m, 1H), 7 ,10 (d,

J=5.0 Hz, 1H), 6.74 (d, J=8.3 Hz, 1H), 6.69 (t, J=8.8 Hz, 1H), 4.76-4.98 ( m, 1H), 4.15-4.31 (m, 1H), 3.99 (br s,

1H), 3.78-3.89 (m, 1H), 3.55-3.77 (m, 1H), 2.99-3.29 (m, 2H), 1.91 (d, J= 2.7 Hz, 3H), 1.68 (td, J=8.0, 4.5 Hz,

1H), 1.45 (s, 9H), 1.35 (dd, J=18.7, 6.6 Hz, 3H), 0.82-0.89 (m, 1H), 0.71-0 .82 (m, 2H), 0.57-0.66 (m, 1H).

Mass/charge (ESI, +ve ion): 605.0 (M+H) ⁺ .

Step 7. 1-(2-Cyclopropyl-4-methyl-3-pyridinyl)-6-fluoro-7-(2-fluoro-6-hydroxyphenyl)-4-((28)-2methyl-4-(2-propenoyl )-1-piperazinyl)pyrido[2,3-b]pyrimidin-2(1H)-one.

Trifluoroacetic acid (2.17 mL, 28.1 mmol) was added to a solution of (38)-tert-butyl-4-(1-(2cyclopropyl-4-methylpyridin-3-yl)-6-fluoro-7-(2- fluoro-6-hydroxyphenyl)-2-oxo-1,2-dihydropyrido[2,3b]pyrimidin-4-yl)-3-methylpiperazine-1-carboxylate (850 mg, 1.41 mmol) in DCM (10 ml) . The resulting mixture was stirred at room temperature for 3 hours and then concentrated. The residue was dissolved in DCM (10 ml), cooled to 0°C and treated with DIPEA (1.23 ml, 7.03 mmol), then with acryloyl chloride (0.103 ml, 1.27 mmol) dropwise using a syringe. The reaction mixture was stirred at 0°C for 2 hours, then quenched with water and extracted with DCM (3x). The combined organic layers were dried over anhydrous sodium sulfate and concentrated. The residue was purified by chromatography on silica gel (eluent: 0-80% 3:1 ESA-EH/heptane) to give 1-(2-cyclopropyl-4-methyl-3-pyridinyl)-6-fluoro-7-(2-fluoro -6hydroxyphenyl)-4-((28)-2-methyl-4-(2-propenoyl)-1-piperazinyl)pyrido[2,3-b]pyrimidin-2(1H)-one.

Λ NMR (400 MHz, DMSO-d ₆ ) δ: ppm 8.32 (br t, J=10.l Hz, 1H), 8.28 (d, J=5.0 Hz, 1H), 7.51 -7.60 (m, 1H), 7.27-7.38 (m, 3H), 7.14 (d, J=4.8 Hz, 1H), 6.80-6.92 (m, 1H ), 6.20 (br d, J=16.6 Hz, 1H), 5.69-5.80 (m, 1H), 4.92 (br d, J=1.5 Hz, 1H), 4 ,24-4.46 (m, 2H), 3.97-4.19 (m, 1H), 3.71 (br s, 1H), 3.42-3.66 (m, 1H), 3.053, 30 (m, 1H), 1.97 (s, 3H), 1.65 (br s, 1H), 1.33 (d, J=6.6 Hz, 3H), 0.90 (td, J= 5.4, 2.6 Hz, 1H), 0.80-0.87 (m, 1H), 0.70-0.79 (m, 1H), 0.60-0.70 (m, 1H) .

Mass/charge (ESI, +ve ion): 559.0 (M+H) ⁺ .

Example 43.

6-Chloro-1-(4,6-di(2-propanyl)-5-pyrimidinyl)-7-(2-fluorophenyl)-4-((28)-2-methyl-4-(2-propenoyl)- 1piperazinyl)pyrido[2,3-b]pyrimidin-2(1H)-one

Xantphos Pd G3, “Αγ ⁰¹ ΑΛΑ

An THF. k. t. An

Stage 1 Intermediate U(1) (C0CI) ₂ , DCE, 80 -c

Intermediate connection U

MeCN,k,t,„

connection P

KH MDS

THF, k.t.

MeCN, K.T.

(1) POCIs, DIPEA, MeCN, 80 'C (2) DIPEA,

Stage 4

Intermediate Stage 2

Stage 3

Step 1. 4,6-Diisopropylpyrimidin-5-amine (intermediate U).

A solution of 4,6-dichloro-5-aminopyrimidine (3.00 g, 18.29 mmol, Combi-Blocks Inc., San Diego, CA, USA) in THF (18 ml) was deoxygenated by bubbling argon through the mixture for 5 minutes. 2-Propylzinc bromide (0.5 M solution in THF, 91.0 mL, 45.5 mmol, Sigma-Aldrich, St. Louis, MO) was added via syringe followed by XantPhos Pd G3 (434 mg, 0.46 mmol, Sigma-Aldrich, St. Louis, MO, USA). The resulting mixture was stirred at room temperature for 16 hours and then filtered through a pad of celite. The filter cake was rinsed with EtOAc and the filtrate was collected and concentrated to give 4,6-diisopropylpyrimidin-5-amine (3.45 g). This material was used in the next step without further purification.

Mass/charge (ESI, +ve ion): 180.2 (M+H) ⁺ .

Step 2. 2,5,6-Trichloro-X-((4,6-diisopropylpyrimidin-5-yl)carbamoyl)nicotinamide.

A solution of 2,5,6-trichloronicotinamide (intermediate P, 3.30 g, 14.6 mmol) in 1,2-dichloroethane (49 ml) was treated with oxalyl chloride (2 M solution in DCM, 11.0 ml, 22 .0 mmol). The mixture was heated at 80°C for 45 minutes, then the heating was stopped and the reaction mixture was concentrated. The residue was dissolved in acetonitrile (49 ml), cooled to -10°C and a solution of 4,6-diisopropylpyrimidin-5-amine (intermediate U, 3.15 g, 17.6 mmol) in acetonitrile (5 ml) was added ) using a cannula. The resulting mixture was stirred at room temperature overnight and then concentrated. The residue was suspended in warm 10:1 heptane/EtOAc (110 ml) and filtered. The filtrate was concentrated and the residue was purified by silica gel chromatography (eluent: 0-40% EtOAc/heptane) to give 2,5,6-trichloro-K-((4,6-diisopropylpyrimidin5-yl)carbamoyl)nicotinamide. 1H NMR (400 MHz, DMSO-d ₆ ) δ: ppm 11.30-11.46 (m, 1H), 9.66 (br s, 1H), 8.95-9.01 (m, 1H), 8.65-8.72 (m, 1H), 3.26 (s, 2H), 1.17 (d, J=6.6 Hz, 12 H).

Mass/charge (ESI, +ve ion): 432.0 (M+H) ⁺ .

Step 3. 6,7-Dichloro-1-(4,6-diisopropylpyrimidin-5-yl)pyrido[2,3-d]pyrimidin-2,4(1H,3H)dione.

To a solution of 2,5,6-trichloro-K-((4,6-diisopropylpyrimidin-5-yl)carbamoyl)nicotinamide (2.10 g, 4.9 mmol) in THF (49 ml) at -20°C was added KHMDS (1 M solution in THF, 12.2 ml, 12.2 mmol). The cooling bath was removed and the resulting mixture was stirred for 2 hours at room temperature. The reaction mixture was quenched with saturated aqueous ammonium chloride (50 ml), diluted with brine and extracted with 3:1 EtOAc/MeOH (1x). The layers were separated and the aqueous layer was extracted with additional EtOAc (1x). The combined organic layers were dried over anhydrous magnesium sulfate and concentrated. The residue was suspended in heptane/EtOAc and filtered. The filtrate was concentrated to give 6,7-dichloro-1-(4,6diisopropylpyrimidin-5-yl)pyrido[2,3-d]pyrimidine-2,4(1H,3H)-dione.

1H NMR (400 MHz, DMSO-d6) δ: ppm 12.33 (s, 1H), 9.18 (s, 1H), 8.61 (s, 1H), 2.90-3.02 (m, 2H), 1.10 (d, J=6.6 Hz, 6H), 0.99 (d, J=6.6 Hz, 6H).

Mass/charge (ESI, +ve ion): 394.0 (M+H) ⁺ .

Step 4. (S)-tert-Butyl-4-(6,7-dichloro-1-(4,6-diisopropylpyrimidin-5-yl)-2-oxo-1,2-dihydropyrido[2,3-d] pyrimidin-4-yl)-3-methylpiperazine-1-carboxylate.

To a solution of 6,7-dichloro-1-(4,6-diisopropylpyrimidin-5-yl)pyrido[2,3-d]pyrimidine-2,4(1H,3H)dione (900 mg, 2.28 mmol) and DIPEA (0.518 ml, 2.97 mmol) in acetonitrile (15 ml) was added slowly with phosphorus oxychloride (0.255 ml, 2.74 mmol) using a syringe. The resulting mixture was heated at 80°C for 45 minutes and then cooled to -10°C. DIPEA (1.2 mL, 6.88 mmol) was added followed by cannula addition of a solution of (S)-N-Boc-2-methylpiperazine (1.37 g, 6.85 mmol, Combi-Blocks, Inc., San -Diego, California, USA) in acetonitrile (5 ml). The resulting mixture was warmed to room temperature and stirred for 10 minutes before additional DIPEA (1.2 mL, 6.88 mmol) was added. The reaction mixture was poured into ice water and extracted with EtOAc (2x). The combined organic layers were washed with brine (1x), dried over anhydrous magnesium sulfate and concentrated. The residue was purified by silica gel chromatography (eluent: 0-80% EtOAc/heptane) to give (S)-tert-butyl-4-(6,7-dichloro-1(4,6-diisopropylpyrimidin-5-yl)-2 -oxo-1,2-dihydropyrido[2,3-d]pyrimidin-4-yl)-3-methylpiperazine 1-carboxylate.

1H NMR (400 MHz, DMSO-d6) δ: ppm 9.15 (s, 1H), 8.48 (s, 1H), 5.75 (s, 1H), 4.90 (br s, 1H), 4.21 (br d, J=14.1 Hz, 1H), 3.91-4.06 (m, 1H), 3.83 (br d, J=13.3 Hz, 1H), 3.73 (br t, J=10.6 Hz, 1H), 3.03-3.19 (m, 1H), 2.69 (dq, J=13.4, 6.7 Hz, 2H), 1.45 (s, 9H), 1.31-1.36 (m, 3H), 1.09 (d, J=6.6 Hz, 6H), 1.00 (d, J=6.6 Hz, 6H) .

Mass/charge (ESI, +ve ion): 576.2 (M+H) ⁺ .

Step 5. (S)-tert-Butyl-4-(6-chloro-1-(4,6-diisopropylpyrimidin-5-yl)-7-(2-fluorophenyl)-2-oxo1,2-dihydropyrido[2, 3-d]pyrimidin-4-yl)-3-methylpiperazine-1-carboxylate.

A mixture of (S)-tert-butyl-4-(6,7-dichloro-1-(4,6-diisopropylpyrimidin-5-yl)-2-oxo-1,2-dihydropyrido[2,3-b]pyrimidin- 4-yl)-3-methylpiperazine-1-carboxylate (500 mg, 0.87 mmol) and potassium acetate (426 mg, 4.34 mmol) in 1,4-dioxane (4.3 ml) were deoxygenated by sparging with argon through the mixture for 5 minutes. [1,1'-Bis-(diphenylphosphino)ferrocene]dichloropalladium(II) complexed with dichloromethane (63 mg, 0.087 mmol) was added and the mixture was heated at 90°C for 10 min. A solution of 2-fluorophenylboronic acid (243 mg, 1.735 mmol, Combi-Blocks, Inc., San Diego, CA, USA) in 1,4-dioxane (2 mL) was slowly added, followed by 6 drops of water. The resulting mixture was heated at 90°C for 1 hour. The reaction mixture was absorbed onto a silica gel plug and purified by silica gel chromatography (eluent: 0-8% MeOH/DCM) to give (S)-tert-butyl-4-(6chloro -1-(4,6-diisopropylpyrimidin-5-yl)-7-(2-fluorophenyl)-2-oxo-1,2-dihydropyrido[2,3-d]pyrimidin4-yl)-3-methylpiperazin-1- carboxylate.

1H NMR (400 MHz, DMSO-d ₆ ) δ: ppm 9.05 (s, 1H), 8.46 (s, 1H), 8.15 (s, 1H), 7.49-7.54 (m , 1H), 7.277.32 (m, 1H), 7.12-7.16 (m, 1H), 4.93 (br s, 1H), 4.29 (br d, J=13.9 Hz, 1H), 4.07 (br d, J=4.6 Hz, 1H), 3.85 (br d, J=13.7 Hz, 1H), 3.75 (br t, J=11.0 Hz , 1H), 3.56 (s, 2H), 3.08-3.22 (m, 3H), 2.67-2.78 (m, 2H), 1.45 (s, 9H), 1, 08 (d, J=6.6 Hz, 6H), 0.92 (d, J=6.6 Hz, 6H).

Mass/charge (ESI, +ve ion): 636.2 (M+H) ⁺ .

Step 6. 6-Chloro-1-(4,6-di(2-propanyl)-5-pyrimidinyl)-7-(2-fluorophenyl)-4-((2S)-2-methyl-4-(2propenoyl) -1-piperazinyl)pyrido[2,3-d]pyrimidin-2(1H)-one.

Trifluoroacetic acid (0.176 ml, 2.36 mmol) was added to a solution of (S)-tert-butyl-4-(6-chloro-1

- 143 044387 (4,6-diisopropylpyrimidin-5-yl)-7-(2-fluorophenyl)-2-oxo-1,2-dihydropyrido[2,3^]pyrimidin-4-yl)-3methylpiperazine-1-carboxylate (150 mg, 0.24 mmol) in DCM (2.4 ml). The resulting mixture was heated at 38°C for 2 hours and then concentrated. The residue was dissolved in DCM (2.4 ml), cooled to 0°C and treated with DIPEA (0.494 ml, 2.83 mmol). After 2 minutes, acryloyl chloride (0.019 mL, 0.24 mmol) was added dropwise using a syringe and the reaction mixture was stirred at 0°C for an additional 10 minutes. The reaction mixture was concentrated and the residue was partitioned between saturated aqueous sodium bicarbonate and EtOAc (2x). The combined organic layers were dried over anhydrous magnesium sulfate and concentrated. The residue was purified by silica gel chromatography (eluent: 0-100% EtOAc/heptane, then 0-8% MeOH/DCM) to give 6-chloro-1-(4,6-di(2propanyl)-5-nirimidinyl)-7 -(2-fluorophenyl)-4-((2S)-2-methyl-4-(2-propenoyl)-1-niperazinyl)pyrido[2,3-d]pyrimidin-2(1H)-one.

^1H NMR (400 MHz, DMSO-d6) δ: ppm 9.06 (s, 1H), 8.46-8.52 (m, 1H), 7.48-7.55 (m, 1H), 7 ,26-7.34 (m, 2H), 7.17 (td, J=7.4, 1.6 Hz, 1H), 6.82-6.93 (m, 1H), 6.22 (br d, J=16.6 Hz, 1H), 5.75-5.80 (m, 1H), 5.00 (br s, 1H), 4.31-4.43 (m, 2H), 4, 02-4.21 (m, 1H), 3.81 (br d, J=8.9 Hz, 1H), 3.45-3.70 (m, 1H), 3.10-3.30 (m , 1H), 2.73 (br d, J=6.4 Hz, 2H), 1.36 (d, J=6.6 Hz, 3H), 1.09 (d, J=6.6 Hz, 6H), 0.93 (d, J=6.4 Hz, 6H).

Mass/charge (ESI, +ve ion): 590.2 (M+H) ⁺ .

Example 44.

6-Chloro-1-(2-cyclopropyl-4-methyl-3-niridinyl)-7-(2-fluoro-6-hydroxyphenyl)-4-((2S)-2-methyl-4-(2propenoyl)-1 -piperazinyl)pyrido[2,3-d]pyrimidin-2(1H)-one (1) (COCIJz, THF, 65 °C (2)

Pd(dppf) ₂ Cl2, KOAc,

Intermediate V

Step 1. 2,5,6-Trichloro-N-((2-cyclopropyl-4-methylpyridin-3-yl)carbamoyl)nicotinamide.

To a solution of 2,5,6-trichloronicotinamide (intermediate P, 3.5 g, 15.5 mmol) in THF (30 mL) was added slowly oxalyl chloride (2 M solution in DCM, 11.6 mL, 23.3 mmol) using a syringe. The resulting mixture was heated at 65°C for 2 hours, then the heating was stopped and the reaction mixture was concentrated. The residue was dissolved in THF (30 ml) and treated with a solution of 2-cyclopropyl-4-methylpyridin-3-amine (intermediate T, 2.5 g, 17.1 mmol) in THF (15 ml) using a cannula. The resulting mixture was stirred at room temperature for 2 hours, then quenched with water and extracted with EtOAc (3x). The combined organic layers were dried over anhydrous sodium sulfate and concentrated to give 2,5,6-trichloro-N-((2cyclopropyl-4-methylpyridin-3-yl)carbamoyl)nicotinamide. This material was used in the next step without further purification.

1H NMR (400 MHz, DMSO-d6) δ: ppm 11.30 (br s, 1H), 9.44-9.73 (m, 1H), 8.64 (s, 1H), 8.21 (d , J=5.0 Hz, 1H), 7.07 (d, J=5.0 Hz, 1H), 2.22 (s, 3H), 0.92 (s, 2H), 0.91 (d , J=3.5 Hz, 2H).

Mass/charge (ESI, +ve ion): 400.9 (M+H) ⁺ .

Step 2. 6,7-Dichloro-1-(2-cyclopropyl-4-methylpyridin-3-yl)pyrido[2,3-d]pyrimidin-2,4(1H,3H) dione.

To an ice-cooled solution of 2,5,6-trichloro-N-((2-cyclopropyl-4-methylpyridin-3yl)carbamoyl)nicotinamide (460 mg, 1.15 mmol) in THF (5 mL), KHMDS (1 M) was slowly added solution in THF, 2.30 ml, 2.30 mmol) using a syringe. The resulting mixture was stirred at 0°C for 2 hours, then quenched with saturated aqueous ammonium chloride and extracted with EtOAc (3x). The combined organic layers were dried over anhydrous sodium sulfate and concentrated. The residue was purified by chromatography on silica gel (eluent: 0-40% 3:1

EtOAc-EtOH/heptane) to give 6,7-dichloro-1-(2-cyclopropyl-4-methylpyridin-3-yl)pyrido[2,3d]pyrimidine-2,4(1H,3H)-dione.

^1H NMR (400 MHz, DMSO-d6) δ: ppm 12.26 (br s, 1H), 8.59 (s, 1H), 8.36 (d, J=5.0 Hz, 1H), 7 .19 (d, J=5.0 Hz, 1H), 2.05 (s, 3H), 1.86-1.96 (m, 1H), 0.88-0.95 (m, 1H), 0.79-0.87 (m, 1H), 0.73-0.79 (m, 1H), 0.620.70 (m, 1H).

Mass/charge (ESI, +ve ion): 362.9 (M+H) ⁺ .

Step 3. 4,6,7-Trichloro-1 -(2-cyclopropyl-4-methylpyridin-3-yl)pyrido[2,3-d]pyrimidin-2(1H)-one.

To a solution of 6,7-dichloro-1-(2-cyclopropyl-4-methylpyridin-3-yl)pyrido[2,3-d]pyrimidine2,4(1H,3H)-dione (4.60 g, 12.7 mmol) and DIPEA (3.32 mL, 19.0 mmol) in acetonitrile (25 mL), phosphorus oxychloride (1.77 mL, 19.0 mmol) was added. The resulting mixture was heated at 80°C for 3 hours and then cooled to room temperature and concentrated to give 4,6,7-trichloro-1-(2cyclopropyl-4-methylpyridin-3-yl)pyrido[2,3-d ]pyrimidin-2(1H)-one. This material was used in the next step without further purification.

Mass/charge (ESI, +ve ion): 380.9 (M+H) ⁺ .

Step 4. (S)-tert-Butyl-4-(6,7-dichloro-1-(2-cyclopropyl-4-methylpyridin-3-yl)-2-oxo-1,2dihydropyrido[2,3-d] pyrimidin-4-yl)-3-methylpiperazine-1-carboxylate (intermediate V).

To an ice-cooled solution of 4,6,7-trichloro-1-(2-cyclopropyl-4-methylpyridin-3yl)pyrido[2,3-d]pyrimidin-2(1H)-one (12.7 mmol) in DCM ( 40 mL) DIPEA (11.1 mL, 63.3 mmol) was added followed by (S)-4-N-Boc-2-methylpiperazine (2.54 g, 12.7 mmol, Combi-Blocks, Inc., San Diego, California, USA). The resulting mixture was stirred at 0°C for 1 hour, then quenched with water and extracted with DCM (3x). The combined organic layers were dried over anhydrous sodium sulfate and concentrated. The residue was purified by silica gel chromatography (eluent: 0-50% 3:1 EtOAc-EtOH/heptane) to give (S)-tert-butyl-4(6,7-dichloro-1-(2-cyclopropyl-4-methylpyridine -3-yl)-2-oxo-1,2-dihydropyrido[2,3-d]pyrimidin-4-yl)3-methylpiperazine-1-carboxylate.

¹ H NMR (400 MHz, DMSO-d ₆ ) δ: ppm 8.44 (d, J=7.0 Hz, 1H), 8.33 (d, J=5.0 Hz, 1H), 7.17 (d, J=5.2 Hz, 1H), 4.85 (br s, 1H), 4.16 (br d, J=11.0 Hz, 1H), 3.96 (br dd, J=3 ,4, 2.2 Hz, 1H), 3.82 (br d, J=13.3 Hz, 1H), 3.69 (q, J=12.0 Hz, 1H), 3.19-3, 29 (m, 1H), 3.03-3.19 (m, 1H), 1.95 (d, J=3.9 Hz, 3H), 1.58-1.68 (m, 1H), 1 .45 (s, 9H), 1.32 (dd, J=6.6, 2.5 Hz, 3H), 0.88-0.97 (m, 1H), 0.77-0.86 (m , 1H), 0.70-0.77 (m, 1H), 0.59-0.69 (m, 1H).

Mass/charge (ESI, +ve ion): 544.9 (M+H) ⁺ .

Step 5. (3S)-tert-Butyl-4-(6-chloro-1-(2-cyclopropyl-4-methylpyridin-3-yl)-7-(2-fluoro-6hydroxyphenyl)-2-oxo-1, 2-dihydropyrido[2,3-d]pyrimidin-4-yl)-3-methylpiperazine-1-carboxylate.

Mixture of (S)-tert-butyl-4-(6,7-dichloro-1-(2-cyclopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydropyrido[2,3-d] pyrimidin-4-yl)-3-methylpiperazine-1-carboxylate (intermediate V, 992 mg, 1.82 mmol), (2-fluoro-6-hydroxyphenyl)boronic acid (510 mg, 3.27 mmol, Combi- Blocks, San Diego, CA, USA), potassium acetate (892 mg, 9.09 mmol) and [1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium(P), complexed with dichloromethane (119 mg, 0.15 mmol ) in 1,4-dioxane (16 ml) was degassed with argon and heated at 80°C. After 2 minutes, two drops of water were added to the reaction mixture and the temperature was increased to 90°C. Heating was continued at 90°C for 1 hour and then the reaction mixture was allowed to cool to room temperature. Water was added and the resulting mixture was extracted with EtOAc (3x). The combined organic layers were dried over anhydrous magnesium sulfate and concentrated. The residue was purified by silica gel chromatography (eluent: 0-50% 3:1 EtOAc-EtOH/heptane) to give (3S)-tert-butyl-4-(6-chloro-1-(2-cyclopropyl4-methylpyridinium-3- yl)-7-(2-fluoro-6-hydroxyphenyl)-2-oxo-1,2-dihydropyrido[2,3-d]pyrimidin-4-yl)-3methylpiperazine-1-carboxylate.

¹ H NMR (400 MHz, DMSO-d6) δ: ppm 10.04-10.22 (m, 1H), 8.31-8.45 (m, 1H), 8.23 (d, J=5, 0 Hz, 1H), 7.19-7.31 (m, 1H), 7.09 (br s, 1H), 6.60-6.76 (m, 2H), 4.75-5.02 ( m, 1H), 4.10-4.36 (m, 1H), 3.92-4.06 (m, 1H), 3.56-3.89 (m, 2H), 2.91-3, 30 (m, 2H), 1.90 (br d, J=19.3 Hz, 3H), 1.61-1.77 (m, 1H), 1.45 (s, 9H), 1.30- 1.40 (m, 3H), 0.59-0.90 (m, 4H).

Mass/charge (ESI, +ve ion): 620.9 (M+H) ⁺ .

Step 6. 1-(2-Cyclopropyl-4-methyl-3-pyridinyl)-6-chloro-7-(2-fluoro-6-hydroxyphenyl)-4-((25)-2methyl-4-(2-propenoyl )-1-piperazinyl)pyrido[2,3-d]pyrimidin-2(1H)-one.

Trifluoroacetic acid (2.47 mL, 33.2 mmol) was added to a solution of (3S)-tert-butyl-4-(6-chloro-1(2-cyclopropyl-4-methylpyridin-3-yl)-7-(2 -fluoro-6-hydroxyphenyl)-2-oxo-1,2-dihydropyrido[2,3d]pyrimidin-4-yl)-3-methylpiperazine-1-carboxylate (1.03 g, 1.66 mmol) in DCM ( 10 ml). The resulting mixture was stirred at room temperature for 3 hours and then concentrated. The residue was dissolved in DCM (10 ml), cooled to 0°C and treated with DIPEA (1.45 ml, 8.29 mmol), then with acryloyl chloride (0.120 ml, 1.49 mmol) dropwise using a syringe. Reactionary

- 145 044387 the mixture was stirred at 0°C for 2 hours, then quenched with water and extracted with

DCM (3x). The combined organic layers were dried over anhydrous sodium sulfate and concentrated. The residue was purified by chromatography on silica gel (eluent: 0-50% 3:1

EtOAc-EtOH/heptane) to give 6-chloro-1-(2-cyclopropyl-4-methyl-3-pyridinyl)-7-(2-fluoro-6hydroxyphenyl)-4-((25)-2-methyl-4 -(2-propenoyl)-1-piperazinyl)pyrido[2,3-d]pyrimidin-2(1H)-one.

1H NMR (400 MHz, DMSO-d6) δ: ppm 10.06-10.26 (m, 1H), 8.33-8.49 (m, 1H), 8.23 (d, J=4.8 Hz, 1H), 7.20-7.34 (m, 1H), 7.09 (br d, J=2.1 Hz, 1H), 6.79-6.92 (m, 1H), 6, 63-6.78 (m, 2H), 6.16-6.29 (m, 1H), 5.76 (dd, J=10.4, 2.3 Hz, 1H), 4.77-5, 08 (m, 1H), 4.21-4.47 (m, 2H), 3.98-4.20 (m, 1H), 3.39-3.92 (m, 2H), 2.92- 3.28 (m, 1H), 1.85-1.99 (m, 3H), 1.62-1.79 (m, 1H), 1.34 (br d, J=19.5 Hz, 3H ), 0.74-0.88 (m, 3H), 0.56-0.68 (m, 1H).

Mass/charge (ESI, +ve ion): 574.9 (M+H)+.

Example 45.

6-Chloro-1-(2-cyclopropyl-4-methyl-3-pyridinyl)-7-(2-fluorophenyl)-4-((2S)-2-methyl-4-(2-propenoyl)1-piperazinyl) pyrido[2,3-d]pyrimidin-2(1H)-one

Step 1. (S)-tert-Butyl-4-(6-chloro-1-(2-cyclopropyl-4-methylpyridin-3-yl)-7-(2-fluorophenyl)-2oxo-1,2-dihydropyrido[ 2,3-d]pyrimidin-4-yl)-3-methylpiperazine-1-carboxylate.

Mixture of (S)-tert-butyl-4-(6,7-dichloro-1-(2-cyclopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydropyrido[2,3-d] pyrimidin-4-yl)-3-methylpiperazine-1-carboxylate (intermediate V, 500 mg, 0.92 mmol), (2-fluorophenyl)boronic acid (269 mg, 1.92 mmol, Combi-Blocks, San- Diego, California, USA), potassium acetate (450 mg, 4.58 mmol) and [1,1-bis-(diphenylphosphino)ferrocene]dichloropalladium(H), complexed with dichloromethane (75 mg, 0.09 mmol) in 1 ,4-dioxane (7 ml) was degassed with argon and heated at 80°C. After 2 min. Three drops of water were added to the reaction mixture and the temperature was raised to 90°C. Heating was continued at 90°C for 1 hour and then the reaction mixture was allowed to cool to room temperature. Water was added and the resulting mixture was extracted with EtOAc (3x). The combined organic layers were dried over anhydrous magnesium sulfate and concentrated. The residue was purified by silica gel chromatography (eluent: 0-40% 3:1 EtOAc-EtOH/heptane) to give (S)-tert-butyl-4-(6-chloro-1-(2-cyclopropyl-4-methylpyridinium- 3yl)-7-(2-fluorophenyl)-2-oxo-1,2-dihydropyrido[2,3-d]pyrimidin-4-yl)-3-methylpiperazine-1carboxylate.

¹ H NMR (400 MHz, DMSO-d ₆ ) δ: ppm 8.41 (d, J=2.5 Hz, 1H), 8.25 (d, J=5.0 Hz, 1H), 7.49 -7.56 (m, 1H), 7.25-7.35 (m, 3H), 7.11 (d, J=4.8 Hz, 1H), 4.90 (br d, J=1, 5 Hz, 1H), 4.24 (br d, J=13.7 Hz, 1H), 3.93-4.07 (m, 1H), 3.85 (br d, J=13.9 Hz, 1H), 3.65-3.79 (m, 1H), 3.21-3.30 (m, 1H), 3.09-3.20 (m, 1H), 1.96 (s, 3H) , 1.601.70 (m, 1H), 1.45 (s, 9H), 1.36 (d, J=6.6 Hz, 3H), 0.86-0.93 (m, 1H), 0, 72-0.83 (m, 2H), 0.61-0.71 (m, 1H).

Mass/charge (ESI, +ve ion): 605.0 (M+H)+.

Step 2. 6-Chloro-1-(2-cyclopropyl-4-methyl-3-pyridinyl)-7-(2-fluorophenyl)-4-((2S)-2-methyl-4-(2pronenoyl)-1- pinerasinyl)pyrido[2,3-d]pyrimidin-2(1H)-one.

Trifluoroacetic acid (1.28 mL, 17.2 mmol) was added to a solution of (S)-tert-butyl-4-(6-chloro-1(2-cyclopropyl-4-methylpyridin-3-yl)-7-(2 -fluorophenyl)-2-oxo-1,2-dihydropyrido[2,3-d]pyrimidin-4yl)-3-methylpiperazine-1-carboxylate (521 mg, 0.86 mmol) in DCM (10 ml). The resulting mixture was stirred at room temperature for 3 hours and then concentrated. The residue was dissolved in DCM (10 ml), cooled to 0°C and treated with DIPEA (0.762 ml, 4.31 mmol), then with acryloyl chloride (0.440 ml, 0.86 mmol) dropwise using a syringe. The reaction mixture was stirred at 0°C for 2 hours, then quenched with water and extracted with DCM (1x). The organic layer was dried over anhydrous sodium sulfate and concentrated. The residue was purified by silica gel chromatography (eluent: 0-50% 3:1 EtOAc-EtOH/heptane) to give 6-chloro-1-(2-cyclopropyl-4-methyl-3-pyridinyl)-7-(2-fluorophenyl) )-4-((2S)-2-methyl-4-(2-propenoyl)-1piperazinyl)pyrido[2,3-d]pyrimidin-2(1H)-one.

1H NMR (400 MHz, DMSO-d6) δ: ppm 8.44 (br s, 1H), 8.25 (d, J=5.0 Hz, 1H), 7.48-7.56 (m, 1H ), 7.247.35 (m, 3H), 7.11 (d, J=5.0 Hz, 1H), 6.78-6.92 (m, 1H), 6.20 (br d, J=16 .2 Hz, 1H), 5.72-5.79 (m, 1H), 4.94 (br s, 1H), 4.25-4.44 (m, 2H), 3.99-4.20 (m, 1H), 3.43-3.69 (m, 2H), 3.02-3.15 (m, 1H), 1.96 (d, J=3.3 Hz, 3H), 1, 60-1.71 (m, 1H), 1.34 (d, J=6.8 Hz, 3H), 0.89 (br dd, J=8.3, 4.6 Hz, 1H), 0. 72-0.84 (m, 2H), 0.61-0.71 (m, 1H).

Mass/charge (ESI, +ve ion): 559.0 (M+H)+.

Example 46.

6-Chloro-1-(2-ethyl-4-methyl-3-pyridinyl)-7-(2-fluorophenyl)-4-((2S)-2-methyl-4-(2-propenoyl)-1piperazinyl)pyrido [2,3-d]pyrimidine-2(1H)-oh.

- 146 044387

Step 1: 2-Ethyl-4-methylpyridin-3-amine (intermediate W).

Ethylmagnesium bromide (3 M solution in diethyl ether, 3.5 ml, 10.5 mmol) was slowly added to a solution of zinc chloride (0.5 M in THF, 18 ml, 9.0 mmol) using a syringe. The addition was exothermic. The solution was stirred at room temperature for 10 min and then 3-amino-2-bromo-4-picoline (1.5 g, 8.0 mmol, Combi-Blocks, San Diego, CA, USA) and the dichloro adduct were added [1,1'-bis-(diphenylphosphino)ferrocene]palladium(P)dichloride with dichloromethane (120 mg, 0.16 mmol). The resulting mixture was heated at 50°C for 20 minutes, then the heating was stopped and the reaction mixture was cooled to room temperature. The reaction mixture was quenched with 10% ammonium hydroxide (30 ml) and extracted with EtOAc (80 ml, 2x40 ml). The combined organic layers were dried over anhydrous sodium sulfate and concentrated. The residue was purified by silica gel chromatography (eluent: 1-5% MeOH/DCM) to give 2-ethyl-4-methylpyridin-3-amine.

1H NMR (400 MHz, CHLOROFORM-d) δ: ppm 7.92 (d, J=5.0 Hz, 1H), 6.87 (d, J=4.8 Hz, 1H), 3.59 (br s, 2H), 2.74 (q, J=7.6 Hz, 2H), 2.19 (s, 3H), 1.33 (t, J=7.6 Hz, 3H).

Mass/charge (ESI, +ve ion): 137.1 (M+H) ⁺ .

Step 2. 2,5,6-Trichloro-N-((2-ethyl-4-methylpyridin-3-yl)carbamoyl)nicotinamide.

To a solution of 2,5,6-trichloronicotinamide (intermediate P, 2.5 g, 11.1 mmol) in THF (20 ml) was added slowly oxalyl chloride (2 M solution in DCM, 5.4 ml, 10.8 mmol) using a syringe. The resulting mixture was heated at 65°C for 1.5 hours, then the heating was stopped and the reaction mixture was cooled to room temperature. A solution of 2-ethyl-4methylpyridin-3-amine (intermediate W, 1.5 g, 10.7 mmol) in THF (15 ml) was added via cannula. The resulting mixture was stirred at room temperature for 1 hour and then partially concentrated to remove most of the THF. The residue was partitioned between saturated aqueous sodium bicarbonate (30 ml) and EtOAc (50 ml). The organic layer was washed with brine (1x), dried over anhydrous sodium sulfate and concentrated. The residue was suspended in 10:1 heptane/EtOAc (60 ml) and filtered to give 2,5,6-trichloro-N-((2-ethyl-4-methylpyridin-3yl)carbamoyl)nicotinamide. The product was carried directly to the next stage.

Step 3. 6,7-Dichloro-1-(2-ethyl-4-methylpyridin-3-yl)pyrido[2,3-d]pyrimidine-2,4(1H,3H)-dione.

To an ice-cooled solution of 2,5,6-trichloro-N-((2-ethyl-4-methylpyridin-3-yl)carbamoyl)nicotinamide (10.7 mmol) in THF (60 ml), KHMDS (1 M solution) was slowly added in THF, 16.0 ml, 16.0 mmol) using a syringe. The resulting solution was stirred at 0°C for 10 min, then quenched with saturated aqueous ammonium chloride (20 ml) and extracted with EtOAc (60 ml). The organic layer was washed with saline (1x), dried over

- 147 044387 anhydrous sodium sulfate and concentrated. The residue was suspended in 9:1 heptane/EtOAc (60 ml) and filtered to give 6,7-dichloro-1-(2-ethyl-4-methylpyridin-3-yl)pyrido[2,3-d]pyrimidine2,4( 1H,3H)-dione.

^1H NMR (400 MHz, DMSO-d6) δ: ppm 12.14-12.39 (m, 1H), 8.54 (s, 1H), 8.47 (d, J=5.0 Hz, 1H ), 7.28 (d, J=5.0 Hz, 1H), 2.41-2.49 (m, 2H), 2.03 (s, 3H), 1.07 (t, J=7, 5 Hz, 3H).

Mass/charge (ESI, +ve ion): 350.9 (M+H) ⁺ .

Step 4. (S)-tert-Butyl-4-(6,7-dichloro-1-(2-ethyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydropyrido[2,3- d]pyrimidin-4-yl)-3-methylpiperazine-1-carboxylate.

To a solution of 6,7-dichloro-1-(2-ethyl-4-methylpyridin-3-yl)pyrido[2,3-d]pyrimidin-2,4(1H,3H)-dione (1.0 g, 2 .9 mmol) and DIPEA (1.5 ml, 8.6 mmol) in acetonitrile (8 ml), phosphorus oxychloride (0.40 ml, 4.3 mmol) was added. The resulting solution was heated at 80°C for 30 minutes and then cooled to 0°C. DIPEA (1.5 mL, 8.6 mmol) was added followed by (S)-4-N-Boc-2-methylpiperazine (800 mg, 4.0 mmol, Combi-Blocks, Inc., San Diego, CA , USA). The resulting mixture was warmed to room temperature and stirred for 30 min, then diluted with EtOAc (30 ml) and washed with saturated aqueous sodium bicarbonate (10 ml) and brine (10 ml). The organic layer was dried over anhydrous magnesium sulfate and concentrated. The residue was purified by silica gel chromatography (eluent: 10-50% 3:1 EtOAc-EtOH/heptane) to give (S)-tert-butyl-4-(6,7-dichloro-1-(2-ethyl-4- methylpyridin-3-yl)-2-oxo-1,2-dihydropyrido[2,3-d]pyrimidin-4-yl)-3-methylpiperazine-1-carboxylate.

A NMR (400 MHz, DMSO-d6) δ: ppm 8.46 (d, J=5.0 Hz, 1H), 8.44 (s, 1H), 7.27 (d, J=5.0 Hz , 1H), 4.85 (br s, 1H), 4.12-4.21 (m, 1H), 3.95 (br s, 1H), 3.82 (br d, J=12.9 Hz , 1H), 3.69 (br d, J=11.4 Hz, 1H), 2.94-3.27 (m, 2H), 2.25-2.43 (m, 2H), 1.94 (d, J=1.7 Hz, 3H), 1.45 (s, 9H), 1.32 (t, J=6.2 Hz, 3H), 1.05 (t, J=7.6 Hz , 3H).

Mass/charge (ESI, +ve ion): 533.0 (M+H) ⁺ .

Step 5. (S)-tert-Butyl-4-(6-chloro-1-(2-ethyl-4-methylpyridin-3-yl)-7-(2-fluorophenyl)-2-oxo-1,2dihydropyrido[ 2,3-d]pyrimidin-4-yl)-3-methylpiperazine-1-carboxylate.

A mixture of (S)-tert-butyl-4-(6,7-dichloro-1-(2-ethyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydropyrido[2,3d]pyrimidin- 4-yl)-3-methylpiperazine-1-carboxylate (450 mg, 0.84 mmol), (2-fluorophenyl)boronic acid (199 mg, 1.43 mmol, Combi-Blocks, San Diego, CA, USA) , potassium acetate (250 mg, 2.55 mmol) and [1,1'-bis-(diphenylphosphino)ferrocene]dichloropalladium(P), complexed with dichloromethane (33 mg, 0.04 mmol) in 1,4-dioxane ( 5 ml) and water (1 ml) were degassed with nitrogen and heated at 65°C for 1.5 hours. Allow the reaction mixture to cool to room temperature, then dilute it with EtOAc (40 ml) and wash with saturated aqueous sodium bicarbonate solution (10 ml) and saline solution (10 ml). The organic layer was dried over anhydrous magnesium sulfate and concentrated. The residue was purified by silica gel chromatography (eluent: 10-50% 3:1 EtOAc-EtOH/heptane) to give (S)-tert-butyl-4-(6-chloro-1-(2-ethyl-4-methylpyridinium- 3-yl)7-(2-fluorophenyl)-2-oxo-1,2-dihydropyrido[2,3-d]pyrimidin-4-yl)-3-methylpiperazine-1-carboxylate.

A NMR (400 MHz, DMSO-d ₆ ) δ: ppm 8.43 (s, 1H), 8.36 (d, J=5.0 Hz, 1H), 7.47-7.55 (m, 1H ), 7.20-7.33 (m, 4H), 4.90 (br s, 1H), 4.24 (br s, 1H), 3.99 (br s, 1H), 3.85 (br d, J=13.3 Hz, 1H), 3.72 (br d, J=11.6 Hz, 1H), 3.33-3.47 (m, 1H), 3.03-3.21 ( m, 1H), 2.30-2.43 (m, 2H), 1.95 (s, 3H), 1.46 (s, 9H), 1.36 (t, J=6.1 Hz, 3H ), 1.04 (dt, J=7.5, 2.3 Hz, 3H).

¹⁹ F NMR (377 MHz, CHLOROFORM-d) δ: ppm -113.84 - -113.85 (2s, 1F).

Mass/charge (ESI, +ve ion): 592.9 (M+H) ⁺ .

Step 6. 6-Chloro-1-(2-ethyl-4-methyl-3-pyridinyl)-7-(2-fluorophenyl)-4-((2S)-2-methyl-4-(2propenoyl)-1 - piperazinyl)pyrido[2,3-d]pyrimidin-2(1H)-one.

Trifluoroacetic acid (3.0 mL, 40.3 mmol) was added to a solution of (S)-tert-butyl-4-(6-chloro-1-(2ethyl-4-methylpyridin-3-yl)-7-(2- fluorophenyl)-2-oxo-1,2-dihydropyrido[2,3-d]pyrimidin-4-yl)-3methylpiperazine-1-carboxylate (500 mg, 0.84 mmol) in DCM (10 ml). The resulting mixture was stirred at room temperature for 30 minutes and then concentrated. The residue was dissolved in DCM (10 ml) and treated with DIPEA (0.500 ml, 2.86 mmol), then with a solution of acryloyl chloride (0.100 ml, 1.23 mmol) in DCM (2 ml). The reaction mixture was stirred at room temperature for 30 minutes and then quenched with saturated aqueous sodium bicarbonate (10 ml) and brine (5 ml). The mixture was extracted with EtOAc (2x15 ml), the combined organic layers were dried over anhydrous sodium sulfate and concentrated. The residue was purified by silica gel chromatography (eluent: 10-60% 3:1 EtOAc-EtOH/heptane) to give 6-chloro-1-(2ethyl-4-methyl-3-pyridinyl)-7-(2-fluorophenyl)- 4-((2S)-2-methyl-4-(2-propenoyl)-1-piperazinyl)pyrido[2,3-d]pyrimidin-2(1H)-one.

A NMR (400 MHz, CHLOROFORM-d) δppm 8.45 (d, J=5.0 Hz, 1H), 8.10 (s, 1H), 7.38-7.46 (m, 1H), 7 .08-7.20 (m, 4H), 6.62 (br s, 1H), 6.42 (dd, J=16.7, 1.6 Hz, 1H), 5.82 (dd, J= 10.5, 1.8 Hz, 1H), 4.24-5.21 (m, 3H), 3.60-4.15 (m, 3H), 3.00-3.36 (m, 1H) , 2.41-2.60 (m, 2H), 2.00-2.12 (m, 3H), 1.45-1.61 (m, 3H), 1.15-1.22 (m, 3H).

¹⁹ F NMR (377 MHz, CHLOROFORM-d) δ: ppm -111.94 - -113.08 (m, 1F).

- 148 044387

Mass/charge (ESI, +ve ion): 547.2 (M+H) ⁺ .

Example 47.

6-Chloro-1-(4,6-diethyl-5-pyrimidinyl)-7-(2-fluorophenyl)-4-((2S)-2-methyl-4-(2-propenoyl)-1piperazinyl)pyrido [2 ,3-d]pyrimidin-2(1H)-one

Pd(dppf)CI ₂ Cs ₂ CO ₃ , nh ₂

NH ₂

H ₂ , Pd/C nh ₂ dioxane/H ₂ O, 100°C

Stage 1

EtOH. K.T.

Stage 2

Intermediate X (1) (COCI) ₂ , THF, 65 °C

(1) P0CI ₃ , TEA, MeCN, 60 °C (2) TEA,

Step 1. 4,6-Dvinylpyrimidin-5-amine.

A mixture of 4,6-dichloro-5-aminopyrimidine (5.0 g, 30.5 mmol, Sigma Aldrich, St. Louis, MO, USA), potassium vinyl trifluoroborate (16.3 g, 122 mmol, Sigma Aldrich, St. Louis , Missouri, USA), Pd(dppf)Cl ₂ (1.12 g, 1.52 mmol) and cesium carbonate (49.7 g, 152 mmol) in 1,4-dioxane (130 ml) and water (13 ml ) was degassed with nitrogen and heated at 100°C for 4 hours. The reaction mixture was allowed to cool to room temperature and then diluted with water and extracted sequentially with EtOAc (3x100 ml) and DCM (5x100 ml). The combined organic layers were dried over anhydrous magnesium sulfate and concentrated. The residue was purified by silica gel chromatography (eluent: 0-20% 4:1 DCM-MeOH/DCM) to give 4,6-divinylpyrimidin-5 amine.

¹ H NMR (400 MHz, CHLOROFORM-d) δ: ppm 8.62 (s, 1H), 6.80-6.89 (m, 2H), 6.46 (dd, J=17.0, 1, 7 Hz, 2H), 5.71 (dd, J=10.9, 1.8 Hz, 2H), 3.86 (br s, 2H).

Mass/charge (ESI, +ve ion): 148.1 (M+H) ⁺ .

Step 2: 4,6-Diethylpyrimidin-5-amine (Intermediate X).

A solution of 4,6-divinylpyrimidin-5-amine (3.1 g, 21.1 mmol) in ethanol (50 ml) was treated with palladium (10 wt.% on activated carbon, 1.12 g, 1.05 mmol) . The mixture was purged with hydrogen at 20 psi. inch (4x) and stirred under a hydrogen atmosphere at 20 psi. inch at room temperature for 4 hours. The reaction mixture was filtered through a pad of celite and concentrated to give 4,6-diethylpyrimidin-5-amine.

1H NMR (400 MHz, CHLOROFORM-d) δ: ppm 8.59 (s, 1H), 3.62 (br s, 2H), 2.70 (q, J=7.6 Hz, 4H), 1, 34 (t, J=7.6 Hz, 6H).

Mass/charge (ESI, + ve ion): 152.2 (M+H) ⁺ .

Step 3. 2,5,6-Trichloro-N-((4,6-diethylpyrimidin-5-yl)carbamoyl)nicotinamide.

To a suspension of 2,5,6-trichloronicotinamide (intermediate P, 4.67 g, 20.7 mmol) in THF (100 mL) was added oxalyl chloride (2 M solution in DCM, 15.5 mL, 31.0 mmol). The resulting mixture was heated at 65°C for 1 hour, then the heating was stopped and the reaction mixture was concentrated to one third of the volume. Toluene (100 ml) was added and the mixture was cooled to 0°C. A solution of 4,6-diethylpyrimidin-5-amine (intermediate X, 3.13 g, 20.7 mmol) in 1,2-dichloroethane (20 ml) was added dropwise via cannula. The resulting mixture was heated to room temperature and stirred in

- 149 044387 for 15 minutes and then concentrated. The residue was suspended in MTBE (50 ml) and filtered. The filtered solids were briefly dried in a vacuum oven at 50° C. to obtain 2,5,6-trichloro-K-((4,6-diethylpyrimidin-5-yl)carbamoyl)nicotinamide.

Mass/charge (ESI, +ve ion): 402.1 (M+H) ⁺ .

Step 4. 6,7-Dichloro-1-(4,6-diethylpyrimidin-5-yl)pyrido[2,3-d]pyrimidine-2,4(1H,3H)-dione.

KHMDS ( 1 M solution in THF, 36.2 ml, 36.2 mmol) using a syringe. The resulting mixture was warmed to room temperature and stirred for 5 min, then quenched with saturated aqueous ammonium chloride and extracted with EtOAc (5x100 ml). The combined organic layers were dried over anhydrous magnesium sulfate and concentrated. The residue was dried overnight in a vacuum oven at 45°C to give 6,7-dichloro-1-(4,6-diethylpyrimidin-5-yl)pyrido[2,3-d]pyrimidine-2,4(1H,3H )dione. This material was used in the next step without further purification.

Mass/charge (ESI, +ve ion): 366.0 (M+H) ⁺ .

Step 5. (S)-tert-Butyl-4-(6,7-dichloro-1-(4,6-diethylpyrimidin-5-yl)-2-oxo-1,2-dihydropyrido[2,3-d] pyrimidin-4-yl)-3-methylpiperazine-1-carboxylate.

Solution of 6,7-dichloro-1-(4,6-diethylpyrimidin-5-yl)pyrido[2,3-d]pyrimidine-2,4(1H,3H)-dione (3.87 g, 10.6 mmol ) in acetonitrile (30 ml) was treated with triethylamine (3.71 ml, 26.4 mmol), then with phosphorus oxychloride (1.26 ml, 12.7 mmol). The resulting mixture was heated at 60°C for 30 minutes and then cooled to 0°C. Triethylamine (1.50 mL, 10.7 mmol) was added followed by (S)-S-N-Boc-2-methylpiperazine (2.22 g, 11.1 mmol, Combi-Blocks, Inc., San Diego, CA, USA ). The reaction mixture was warmed to room temperature and stirred for 30 min, then partitioned between EtOAc (100 ml) and brine (40 ml). The organic layer was washed with water (30 ml) and brine (50 ml), dried over anhydrous magnesium sulfate and concentrated. The residue was purified by silica gel chromatography (eluent: 0100% EtOAc/heptane) to give (S)-tert-butyl-4-(6,7-dichloro-1-(4,6-diethylpyrimidin-5-yl)-2- oxo1,2-dihydropyrido[2,3-d]pyrimidin-4-yl)-3-methylpiperazine-1-carboxylate.

1H NMR (400 MHz, DMSO-d6) δ: ppm 9.09 (s, 1H), 8.48 (s, 1H), 4.88 (br s, 1H), 4.19 (br d, J= 13.7 Hz, 1H), 3.90-4.00 (m, 1H), 3.83 (br d, J=13.7 Hz, 1H), 3.66-3.78 (m, 1H) , 3.05-3.35 (m, 2H), 2.35-2.46 (m, 4H), 1.45 (s, 9H), 1.33 (d, J=6.6 Hz, 3H ), 1.08 (t, J=7.5 Hz, 6H).

Mass/charge (ESI, +ve ion): 548.2 (M+H) ⁺ .

Step 6. (S)-tert-Butyl-4-(6-chloro-1-(4,6-diethylpyrimidin-5-yl)-7-(2-fluorophenyl)-2-oxo-1,2dihydropyrido[2, 3-d]pyrimidin-4-yl)-3-methylpiperazine-1-carboxylate.

A mixture of (S)-tert-butyl-4-(6,7-dichloro-1-(4,6-diethylpyrimidin-5-yl)-2-oxo-1,2-dihydropyrido[2,3d]pyrimidin-4- yl)-3-methylpiperazine-1-carboxylate (354 mg, 0.65 mmol), (2-fluorophenyl)boronic acid (108 mg, 0.78 mmol, Combi-Blocks, San Diego, CA, USA), acetate potassium (317 mg, 3.23 mmol) and [1,1'-bis-(diphenylphosphino)ferrocene]dichloropalladium(P), complexed with dichloromethane (23 mg, 0.03 mmol) in 1,4-dioxane (6, 5 ml) was degassed with argon for 5 min. Four drops of water were added and the mixture was heated at 90°C for 30 minutes. The reaction mixture was allowed to cool to room temperature and then partitioned between water (10 ml) and EtOAc (20 ml). The aqueous layer was extracted with DCM (20 ml). The combined organic layers were washed with brine (20 ml), dried over anhydrous magnesium sulfate and concentrated. The residue was purified by silica gel chromatography (eluent: 0-30% 4:1 DCM-MeOH/DCM) to give (S)-tert-butyl-4-(6-chloro-1-(4,6-diethylpyrimidine-5- yl)-7-(2-fluorophenyl)-2-oxo-1,2-dihydropyrido[2,3-d]pyrimidin-4-yl)-3-methylpiperazine-1-carboxylate.

1H NMR (400 MHz, DMSO-d ₆ ) δ: ppm 9.01 (s, 1H), 8.45 (s, 1H), 7.49-7.57 (m, 1H), 7.26-7 .34 (m, 2H), 7.20-7.26 (m, 1H), 4.88-4.98 (m, 1H), 4.27 (br d, J=13.7 Hz, 1H) , 3.93-4.03 (m, 1H), 3.85 (br d, J=13.5 Hz, 1H), 3.70-3.81 (m, 1H), 3.05-3, 35 (m, 2H), 2.35-2.46 (m, 4H), 1.46 (s, 9H), 1.37 (d, J=6.6 Hz, 3H), 1.06 (td , J=7.5, 1.6 Hz, 6H).

¹⁹ F NMR (377 MHz, DMSO-d ₆ ) δ: ppm -114.01 (s, 1F).

Mass/charge (ESI, +ve ion): 608.2 (M+H) ⁺ .

Step 7. 6-Chloro-1-(4,6-diethyl-5-pyrimidinyl)-7-(2-fluorophenyl)-4-((2S)-2-methyl-4-(2-propenoyl)1-piperazinyl )pyrido[2,3-d]pyrimidin-2(1H)-one.

Trifluoroacetic acid (3.0 mL, 38.9 mmol) was added to a solution of (S)-tert-butyl-4-(6-chloro-1(4,6-diethylpyrimidin-5-yl)-7-(2-fluorophenyl )-2-oxo-1,2-dihydropyrido[2,3-d]pyrimidin-4-yl)-3methylpiperazine-1-carboxylate (327 mg, 0.54 mmol) in DCM (5 ml). The resulting mixture was stirred at room temperature for 15 minutes and then concentrated. The residue was dissolved in DCM (10 ml), cooled to 0°C and treated with DIPEA (0.470 ml, 2.69 mmol), then with acryloyl chloride (0.048 ml, 0.59 mmol). The reaction mixture was stirred at 0°C for 10 min and then additional DIPEA (0.300 mL, 1.71 mmol) was added, followed by additional acryloyl chloride (0.020 mL, 0.25 mmol). After an additional 20 minutes at

- 150 044387

The 0°C reaction mixture was quenched with saturated aqueous sodium bicarbonate (10 ml) and extracted with DCM (2x30 ml). The combined organic layers were dried over anhydrous magnesium sulfate and concentrated. The residue was purified by silica gel chromatography (eluent: 0-60% 4:1 DCM-MeOH/DCM) to give 6-chloro-1-(4,6-diethyl-5-pyrimidinyl)-7-(2-fluorophenyl)- 4((2S)-2-methyl-4-(2-propenoyl)-1-piperazinyl)pyrido[2,3-d]pyrimidin-2(1H)-one.

^1H NMR (400 MHz, DMSO-d6) δ: ppm 9.01 (s, 1H), 8.48 (br d, J=5.0 Hz, 1H), 7.50-7.57 (m, 1H), 7.277.35 (m, 3H), 7.20-7.27 (m, 1H), 6.79-6.93 (m, 1H), 6.17-6.26 (m, 1H) , 5.65-5.80 (m, 1H), 4.92-5.02 (m, 1H), 4.23-4.44 (m, 2H), 3.99-4.20 (m, 1H), 3.72-3.88 (m, 1H), 3.40-3.71 (m, 1H), 2.35-2.48 (m, 4H), 1.36 (d, J= 6.6 Hz, 3H), 1.06 (t, J=7.5 Hz, 6H).

¹⁹ F NMR (377 MHz, DMSO-d ₆ ) δ: ppm -114.01 (s, 1F).

Mass/charge (ESI, +ve ion): 562.1 (M+H) ⁺ .

Example 48.

6-Chloro-7-(2-fluoro-6-hydroxyphenyl)-4-((2S)-2-methyl-4-(2-pronoyl)-1-piperazinyl)-1-(4-(2propanyl)-1 ,3-thiazol-5-yl)pyrido[2,3-d]pyrimidin-2(1H)-one (1) (COCI) ₂ , THF, 70 °CC) ^P0CI 3- ^DIPEA >

MeCN, 80 °C 1 (2) DIPEA.

U ri Vos—N NH ⁵ o _h /4 ¹ n,

O THF, k.t. rQ 7 I _Λ a ^KHMDS iHL A ^{CI DM} E ^KT

ΉΓΑοι -------- ν yY ------ уУ ------ ^{H N} cr ^N Stage 1 ci n CI _TH p, kt | Stage 3

Intermediate Stage 2/ ^N connection P

Step 1. 2,5,6-Trichloro-N-((4-isopropylthiazol-5-yl)carbamoyl)nicotinamide.

To a mixture of 2,5,6-trichloronicotinamide (intermediate P, 1.00 g, 4.5 mmol) in THF (20 mL) was added oxalyl chloride (2 M solution in DCM, 2.4 mL, 4.9 mmol). The resulting mixture was heated at 70°C for 30 minutes, then the heating was stopped and the reaction mixture was cooled to room temperature. 4-(propan-2-yl)-1,3-thiazol-5-amine (715 mg, 5.0 mmol, Enamine, Monmouth Junction, NJ, USA) was added. The reaction mixture was stirred at room temperature for 10 minutes and then concentrated. The residue was suspended in MeOH and filtered. The filtered solids were discarded and the filtrate was concentrated to give 2,5,6-trichloro-N((4-isopropylthiazol-5-yl)carbamoyl)nicotinamide.

^1H NMR (400 MHz, DMSO- _d6 ) δ: ppm 11.69 (br s, 1H), 10.64 (br s, 1H), 8.71 (s, 1H), 8.58 (s, 1H), 3.13 (br d, J=6.4 Hz, 1H), 1.25 (d, J=6.8 Hz, 6H).

Mass/charge (ESI, +ve ion): 393.0 (M+H) ⁺ .

Step 2. 6,7-Dichloro-1-(4-isopropylthiazol-5-yl)pyrido[2,3-d]pyrimidine-2,4(1H,3H)-dione.

To an ice-cooled solution of 2,5,6-trichloro-N-((4-isopropylthiazol-5-yl)carbamoyl)nicotinamide (1.75 g, 4.5 mmol) in THF (20 mL) was added KHMDS (1 M solution in THF, 9.3 ml, 9.3 mmol). The resulting mixture was warmed to room temperature and stirred for 30 min, then re-cooled to 0°C and treated with KHMDS (IM solution in THF, 1.0 ml, 1.0 mmol). The reaction mixture was warmed to room temperature and stirred for 15 min, then quenched with semi-saturated aqueous ammonium chloride (60 ml) and extracted with EtOAc (2x50 ml). The combined organic layers were washed with water (60 ml), dried by elution through a Chem Elut extraction thimble (Agilent Technologies, Santa Clara, CA, USA) and concentrated. The residue was purified by silica gel chromatography (eluent: 5-70% EtOAc/heptane) to give 6,7-dichloro-1-(4-isopropylthiazol-5-yl)pyrido[2,3d]pyrimidine-2,4(1H, 3H)-dione.

¹ H NMR (400 MHz, DMSO-d ₆ ) δ: ppm 12.20 (s, 1H), 9.14 (s, 1H), 8.54 (s, 1H), 2.86 (quin, J= 6.8 Hz, 1H), 1.11 (t, J=7.4 Hz, 6H).

Mass/charge (ESI, +ve ion): 357.0 (M+H) ⁺ .

Step 3. (S)-tert-Butyl-4-(6,7-dichloro-1-(4-isopropylthiazol-5-yl)-2-oxo-1,2-dihydropyrido[2,3d]pyrimidin-4- yl)-3-methylpiperazine-1-carboxylate.

To a mixture of 6,7-dichloro-1-(4-isopropylthiazol-5-yl)nirido[2,3-d]pyrimidine-2,4(1H,3H)-dione (3.01 g, 8.4 mmol) in acetonitrile (40 ml), DIPEA (4.40 ml, 25.3 mmol) was added followed by phosphorus oxychloride (1.57 ml, 16.8 mmol). The resulting mixture was heated at 80°C for 15 minutes, then cooled to room temperature and concentrated. The residue was dissolved in DMF (30 ml) and

- 151 044387 were treated with DIPEA (7.34 ml, 42.2 mmol), then with (S)-4-N-Boc-2methylpiperazine (1.89 g, 9.5 mmol, Sigma-Aldrich, St. Louis, Missouri, USA). The resulting solution was stirred at room temperature for 5 minutes, then ice water (100 ml) was added and the mixture was stirred for an additional 15 minutes. The mixture was filtered and the filtered solids were dissolved with DCM, dried by passing through a Chem Elut extraction thimble (Agilent Technologies, Santa Clara, CA, USA) and concentrated. The residue was purified by silica gel chromatography (eluent: 40-100% EtOAc/heptane) to give (S)tert-butyl-4-(6,7-dichloro-1-(4-isopropylthiazol-5-yl)-2-oxo -1,2-dihydropyrido[2,3-d]pyrimidin-4-yl)3-methylpiperazine-1-carboxylate.

1H NMR (400 MHz, DMSO-d ₆ ) δ: ppm 9.10 (s, 1H), 8.40 (d, J=1.7 Hz, 1H), 4.85 (br s, 1H), 4 .07-4.18 (m, 1H), 3.88-4.00 (m, 1H), 3.81 (br d, J=13.9 Hz, 1H), 3.64-3.77 ( m, 1H), 3.28 (s, 1H), 3.01-3.19 (m, 1H), 2.65 (t, J=13.3, 6.6 Hz, 1H), 1.44 (s, 9H), 1.31 (dd, J=6.4, 3.3 Hz, 3H), 1.09-1.13 (m, 6H).

Mass/charge (ESI, +ve ion): 539.2 (M+H) ⁺ .

Step 4. (3S)-tert-Butyl-4-(6-chloro-7-(2-fluoro-6-hydroxyphenyl)-1-(4-isopropylthiazol-5-yl)-2oxo-1,2-dihydropyrido[ 2,3-d]pyrimidin-4-yl)-3-methylpiperazine-1-carboxylate.

Mixture of (s)-tert-butyl-4-(6,7-dichloro-1-(4-isopropylthiazol-5-yl)-2-oxo-1,2-dihydropyrido[2,3d]pyrimidin-4-yl) -3-methylpiperazine-1-carboxylate (2.03 g, 3.76 mmol), potassium acetate (1.86 g, 18.9 mmol), (2-fluoro-6-hydroxyphenyl)boronic acid (911 mg, 5 .84 mmol, Combi-Blocks, San Diego, CA, USA) and [1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium(P), complexed with dichloromethane (282 mg, 0.39 mmol) in 1,4- dioxane (36 ml) was degassed with argon for 5 min. A drop of water was added and the resulting mixture was heated to 90°C for 1.5 hours. The reaction mixture was allowed to cool to room temperature and then partitioned between water (50 ml) and EtOAc (2x50 ml). The combined organic extracts were washed with water (60 mL), dried by elution through a Chem Elut extraction thimble (Agilent Technologies, Santa Clara, CA, USA) and concentrated. The residue was purified by silica gel chromatography (eluent: 50-100% EtOAc/heptane) to give (3S)-tert-butyl-4-(6-chloro-7-(2-fluoro-6-hydroxyphenyl)-1(4- isopropylthiazol-5-yl)-2-oxo-1,2-dihydropyrido[2,3-d]pyrimidin-4-yl)-3-methylpiperazine-1carboxylate.

1H NMR (400 MHz, DMSO-d6) δ: ppm 10.10 (s, 1H), 9.00 (s, 1H), 8.31-8.39 (m, 1H), 7.21-7, 32 (m, 1H), 6.65-6.77 (m, 2H), 4.88 (br s, 1H), 4.11-4.33 (m, 1H), 3.91-3.98 (m, 1H), 3.83 (br d, J=13.9 Hz, 1H), 3.64-3.78 (m, 1H), 3.28 (br s, 1H), 3.03- 3.21 (m, 1H), 2.63 (br s, 1H), 1.45 (s, 9H), 1.36 (br s, 3H), 1.09 (br d, J=6.8 Hz, 3H), 1.01 (d, J=6.8 Hz, 3H).

¹⁹ F NMR (376 MHz, DMSO d6) δ: ppm -115.41 (br s, 1F).

Mass/charge (ESI, +ve ion): 615.2 (M+H) ⁺ .

Step 5. 6-Chloro-7-(2-fluoro-6-hydroxyphenyl)-4-((2S)-2-methyl-4-(2-propenoyl)-1-piperazinyl)-1(4-(2- propanyl)-1,3-thiazol-5-yl)pyrido[2,3-d]pyrimidin-2(1H)-one.

Trifluoroacetic acid (10 ml, 130 mmol) was added to a solution of (3S)-tert-butyl-4-(6-chloro-7-(2fluoro-6-hydroxyphenyl)-1-(4-isopropylthiazol-5-yl)-2 -oxo-1,2-dihydropyrido[2,3-d]pyrimidin-4-yl)-3methylpiperazine-1-carboxylate (2.15 g, 3.50 mmol) in DCM (20 ml). The reaction mixture was stirred for 30 minutes at room temperature and then concentrated. The residue was dissolved in DCM (20 ml), cooled to 0°C and treated with DIPEA (3.05 ml, 17.5 mmol), then with acryloyl chloride solution (0.258 M in DCM, 10.84 ml, 2.8 mmol). The reaction mixture was warmed to room temperature and stirred for 15 min, then re-cooled to 0°C and treated with acryloyl chloride solution (0.258 M in DCM, 2.0 ml, 0.52 mmol). After warming to room temperature and stirring for an additional 10 minutes, the reaction mixture was concentrated. The residue was purified by silica gel chromatography (eluent: 40-100% 3:1 EtOAc-EtOH/heptane) to give 6-chloro-7-(2-fluoro-6-hydroxyphenyl)-4-((2S)-2-methyl -4-(2-propenoyl)1-piperazinyl)-1-(4-(2-propanyl)-1,3-thiazol-5-yl)pyrido[2,3-d]pyrimidin-2(1H)-one .

1H NMR (400 MHz, DMSO-d6) δ: ppm 10.10 (s, 1H), 9.01 (s, 1H), 8.37 (br s, 1H), 7.22-7.32 (m , 1H), 6.80-6.93 (m, 1H), 6.66-6.77 (m, 2H), 6.20 (br d, J=16.6 Hz, 1H), 5.71 -5.81 (m, 1H), 4.83-5.04 (m, 1H), 3.984.45 (m, 3H), 3.56-3.87 (m, 2H), 3.07 (br dd, J=4.1, 2.9 Hz, 1H), 2.59-2.70 (m, 1H), 1.33 (br d, J=5.8 Hz, 3H), 1.07- 1.14 (m, 3H), 1.01 (br d, J=6.6 Hz, 3H).

¹⁹ F NMR (376 MHz, DMSO-d ₆ ) δ: ppm -115.41 (br s, 1F).

Mass/charge (ESI, +ve ion): 569.2 (M+H) ⁺ .

Example 49.

6-Chloro-7-(2-fluoro-6-hydroxyphenyl)-1-(2-(2-methyl-2-propanyl)phenyl)-4-((2S)-2-methyl-4-(2propenoyl)- 1-piperazinyl)pyrido[2,3-d]pyrimidine-2(1H)-oh

- 152 044387

Step 1. N-((2-(tert-Butyl)phenyl)carbamoyl)-2,5,6-trichloronicotinamide.

To a mixture of 2,5,6-trichloronicotinamide (intermediate P, 1.02 g, 4.5 mmol) in THF (20 ml) was added oxalyl chloride (2 M solution in DCM, 2.5 ml, 5.0 mmol). The resulting mixture was heated at 70°C for 40 minutes, then the heating was stopped and the reaction mixture was cooled to room temperature. 2-tert-Butylaniline (0.708 ml, 4.5 mmol, Ark Pharm, Arlington Heights, IL, USA) was added. The reaction mixture was stirred at room temperature for 10 minutes and then concentrated. The residue was suspended in MeOH and filtered. The filtered solids were collected to give N-((2-(tert-butyl)phenyl)carbamoyl)-2,5,6-trichloronicotinamide.

1H NMR (400 MHz, DMSO-d6) δ: ppm 11.43 (br s, 1H), 9.79-10.16 (m, 1H), 8.65 (s, 1H), 7.49 (br dd, J=7.5, 1.7 Hz, 1H), 7.43 (br d, J=7.7 Hz, 1H), 7.18-7.28 (m, 2H), 1.40 ( s, 9H).

Mass/charge (ESI, + ve ion): 422.0 (M+Na) ⁺ .

Step 2. 1-(2-(tert-Butyl)phenyl)-6,7-dichloropyrido[2,3-d]pyrimidine-2,4(1H,3H)-dione.

To an ice-cooled mixture of N-((2-(tert-butyl)phenyl)carbamoyl)-2,5,6-trichloronicotinamide (1.26 g, 3.1 mmol) in THF (20 mL) was added KHMDS (1 M solution in THF, 6.6 ml, 6.6 mmol). The resulting mixture was warmed to room temperature and stirred for 10 min, then quenched by adding semi-saturated aqueous ammonium chloride (60 ml) and extracted with EtOAc (2x50 ml). The combined organic layers were washed with water (60 mL), dried by elution through a Chem Elut extraction thimble (Agilent Technologies, Santa Clara, CA, USA) and concentrated. The residue was suspended in MeOH and filtered. The filtered solids were collected to give 1-(2-(tert-butyl)phenyl)-6,7-dichloropyrido[2,3-d]pyrimidine2,4(1H,3H)-dione.

^1H NMR (400 MHz, DMSO-d6) δ: ppm 12.19 (s, 1H), 8.56 (s, 1H), 7.65 (dd, J=8.2, 1.3 Hz, 1H ), 7.41-7.46 (m, 1H), 7.33 (td, J=7.5, 1.2 Hz, 1H), 7.21 (dd, J=7.8, 1.3 Hz, 1H), 1.17 (s, 9H).

Mass/charge (ESI, +ve ion): 364.0 (M+H) ⁺ .

Step 3. (S)-tert-Butyl-4-(1-(2-(tert-butyl)phenyl)-6,7-dichloro-2-oxo-1,2-dihydropyrido[2,3d]pyrimidine-4 -yl)-3-methylpiperazine-1-carboxylate.

To a mixture of 1-(2-(tert-butyl)phenyl)-6,7-dichloropyrido[2,3-d]pyrimidine-2,4(1H,3H)-dione (925 mg, 2.54 mmol) in acetonitrile (20 ml) DIPEA (0.574 ml, 3.30 mmol) was added followed by phosphorus oxychloride (0.284 ml, 3.05 mmol). The resulting mixture was heated at 80°C for 45 minutes, then cooled to 0°C. DIPEA (1.33 mL, 7.62 mmol) was added followed by (S)-4-N-Boc-2-methylpiperazine (0.522 g, 2.61 mmol, Sigma-Aldrich, St. Louis, MO, USA) . Allow the resulting solution to gradually warm to room temperature over a period of 40 minutes, then pour it into cold saturated aqueous sodium bicarbonate solution (50 ml) and stir for an additional 10 minutes. The mixture was extracted with EtOAc (2x30 ml) and the combined organic extracts were dried by elution through a Chem Elut extraction thimble (Agilent Technologies, Santa Clara, CA, USA) and concentrated. The residue was purified by silica gel chromatography (eluent: 30-80% EtOAc/heptane) to give (S)-tert-butyl-4-(1-(2-(tert-butyl)phenyl)-6,7-dichloro-2 -oxo-1,2-dihydropyrido[2,3-d]pyrimidin-4-yl)3-methylpiperazine-1-carboxylate.

1H NMR (400 MHz, DMSO-d ₆ ) δ: ppm 8.33-8.48 (m, 1H), 7.61 (d, J=7.9 Hz, 1H), 7.36-7.42 (m, 1H), 7.29 (td, J=7.5, 1.2 Hz, 1H), 7.00 (dd, J=7.7, 1.0 Hz, 1H), 4.68- 4.89 (m, 1H), 3.88-4.25 (m, 2H), 3.77-3.86 (m, 1H), 3.52-3.71 (m, 1H), 3, 09-3.26 (m, 1H), 2.84-3.07 (m, 1H), 1.44 (s, 9H), 1.24 (s, 3H), 1.13 (d, J= 2.3 Hz, 9H).

Mass/charge (ESI, +ve ion): 546.2 (M+H) ⁺ .

Step 4. (3S)-tert-Butyl-4-(1-(2-(tert-butyl)phenyl)-6-chloro-7-(2-fluoro-6-hydroxyphenyl)-2-oxo1,2-dihydropyrido [2,3-d]pyrimidin-4-yl)-3-methylniperazine-1-carboxylate.

- 153 044387

Mixture of (S)-tert-butyl-4-(1-(2-(tert-butyl)phenyl)-6,7-dichloro-2-oxo-1,2-dihydropyrido[2,3-d]pyrimidine-4 -yl)-3-methylpiperazine-1-carboxylate (1.00 g, 1.83 mmol), potassium acetate (905 mg, 9.22 mmol), (2-fluoro-6-hydroxyphenyl)boronic acid (347 mg, 2.22 mmol, Combi-Blocks, San Diego, CA, USA) and [1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium(P) complexed with dichloromethane (137 mg, 0.19 mmol) in 1.4 α-dioxane (20 ml) was degassed with argon for 5 min. A drop of water was added and the resulting mixture was heated to 90°C for 1.5 hours. The reaction mixture was allowed to cool to room temperature and then partitioned between water (40 ml) and EtOAc (2x40 ml). The combined organic extracts were washed with water (40 mL), dried by elution through a Chem Elut extraction thimble (Agilent Technologies, Santa Clara, CA, USA) and concentrated. The residue was purified by silica gel chromatography (eluent: 40-100% EtOAc/heptane) to give (3S)-tert-butyl-4-(1-(2-(tert-butyl)phenyl)-6-chloro-7-( 2fluoro-6-hydroxyphenyl)-2-oxo-1,2-dihydropyrido[2,3-d]pyrimidin-4-yl)-3-methylpiperazine-1carboxylate.

¹ H NMR (400 MHz, DMSO-d ₆ ) δ: ppm 10.03-10.09 (m, 1H), 8.31-8.39 (m, 1H), 7.53 (d, J=8 ,1 Hz, 1H), 7.25-7.32 (m, 1H), 7.18-7.24 (m, 2H), 6.94 (br d, J=6.6 Hz, 1H), 6.61-6.73 (m, 2H), 4.80 (br d, J=1.2 Hz, 1H), 4.09-4.25 (m, 1H), 3.92-4.01 (m, 1H), 3.84 (br d, J=12.9 Hz, 1H), 3.63 (br d, J=9.1 Hz, 1H), 2.99-3.26 (m, 2H), 1.45 (s, 9H), 1.29-1.37 (m, 3H), 1.11 (s, 9H).

¹⁹ F NMR (376 MHz, DMSO-d6) δ: ppm -115.35 -115.44 (m, 1F).

Mass/charge (ESI, +ve ion): 622.2 (M+H) ⁺ .

Step 5. 6-Chloro-7-(2-fluoro-6-hydroxyphenyl)-1-(2-(2-methyl-2-propanyl)phenyl)-4-((2S)-2-methyl-4(2 -propenoyl)-1-piperazinyl)pyrido[2,3-d]pyrimidin-2(1H)-one.

Trifluoroacetic acid (5.0 mL, 64.9 mmol) was added to a solution of (3S)-tert-butyl-4-(1-(2-(tert-butyl)phenyl)-6-chloro-7-(2-fluoro-6 -Hydroxyphenyl)-2-oxo-1,2-dihydropyrido[2,3-d]pyrimidin-4-yl)-3methylpiperazine-1-carboxylate (984 mg, 1.58 mmol) in DCM (10 ml). The resulting mixture was stirred at room temperature for 30 minutes and then concentrated. The residue was dissolved in DCM (10 ml), cooled to 0°C and treated with DIPEA (1.38 ml, 7.91 mmol), then with acryloyl chloride solution (0.258 M in DCM, 4.90 ml, 1.27 mmol). The reaction mixture was warmed to room temperature and stirred for 30 min, then re-cooled to 0°C and treated with acryloyl chloride solution (0.258 M in DCM, 1.0 ml, 0.26 mmol). After warming to room temperature and stirring for an additional 5 minutes. the reaction mixture was concentrated. The residue was purified by silica gel chromatography (eluent: 40-100% 3:1 EtOAc-EtOH/heptane) to give 6-chloro-7-(2-fluoro-6-hydroxyphenyl)-1-(2-(2-methyl- 2-propanyl)phenyl)-4-((2S)-2-methyl-4-(2-propenoyl)-1-piperazinyl)pyrido[2,3-d]pyrimidin-2(1H)-one.

1H NMR (400 MHz, DMSO-d6) δ: ppm 10.06 (br d, J=5.4 Hz, 1H), 8.38 (br d, J=12.2 Hz, 1H), 7.54 (d, J=7.7 Hz, 1H), 7.17-7.32 (m, 3H), 6.91-7.02 (m, 1H), 6.83 (br dd, J=10, 2, 16.4 Hz, 1H), 6.61-6.74 (m, 2H), 6.20 (br d, J=16.6 Hz, 1H), 5.72-5.80 (m, 1H), 4.85 (br d, J=2.1 Hz, 1H), 4.07-4.51 (m, 3H), 3.59-3.81 (m, 3H), 1.241.37 ( m, 3H), 1.06-1.13 (m, 9H).

¹⁹ F NMR (376 MHz, DMSO-d ₆ ) δ: ppm -115.42 (m, 1F).

Mass/charge (ESI, +ve ion): 576.2 (M+H) ⁺ .

- 154 044387

Example 50 2( 1 H)-pteridinone (PPh ₃ ) ₄ Pd, K ₂ C0 _;

K ₂ N DME/H ₂ O.90 ^s CH ₂ N f-BuOH, 60 °C Cl THF, 50 ^° C

Stage 1 Stage 2 Stage 3

KHMDS

THF, k.t.

Stage 5 (1) POCI ₃ , DIPEA, MeCN, 80 'C (2) DIPEA,

Intermediate Y

Step 1. Methyl 3-amino-6-chloro-5-(2-fluorophenyl)pyrazine-2-carboxylate.

A mixture of methyl 3-amino-5,6-dichloropyrazine-2-carboxylate (10.0 g, 45 mmol, Ark Pharm, Inc., Arlington Hythe, IL), (2-fluorophenyl)boronic acid (6.93 g, 49.5 mmol, Combi-Blocks, San Diego, CA, USA) and potassium carbonate (13.1 g, 95 mmol) in 10:1 DME/water (220 ml) were degassed with nitrogen for 5 min and then tetrakis(triphenylphosphine)palladium(0) (1.04 g, 0.90 mmol) was added. The reaction mixture was stirred at 90°C for 16 hours, then allowed to cool to room temperature and partitioned between EtOAc (200 ml) and 1N. HCl (200 ml). The organic layer was dried over anhydrous sodium sulfate and concentrated to give methyl 3amino-6-chloro-5-(2-fluorophenyl)pyrazine-2-carboxylate.

Mass/charge (ESI, +ve): 282.1 (M+H) ⁺ .

Step 2. Methyl 3,6-dichloro-5-(2-fluorophenyl)pyrazine-2-carboxylate.

To a solution of methyl 3-amino-6-chloro-5-(2-fluorophenyl)pyrazine-2-carboxylate (22.0 g, 78 mmol) in t-BuOH (220 mL) was added isoamyl nitrite (15.8 mL, 117 mmol) and copper chloride (12.6 g, 94 mmol). The resulting mixture was stirred at 60°C for 16 hours and then partitioned between water (1 L) and EtOAc (2 L). The organic layer was dried over anhydrous sodium sulfate and concentrated. The residue was purified by silica gel chromatography (eluent: 0-5% EtOAc/hexane) to give methyl 3,6-dichloro-5-(2-fluorophenyl)pyrazine-2-carboxylate.

Mass/charge (ESI, +ve ion): 300.9 (M+H) ⁺ .

Step 3. 3,6-Dichloro-5-(2-fluorophenyl)pyrazine-2-carboxamide.

To a solution of methyl 3,6-dichloro-5-(2-fluorophenyl)pyrazine-2-carboxylate (13.5 g, 44.8 mmol) in THF (150 ml) was added a solution of ammonium hydroxide (30%, 150 ml, 44.8 mmol). The resulting mixture was heated at 50°C for 4 hours, then allowed to cool to room temperature and partitioned between water and EtOAc (500 ml). The organic layer was dried over anhydrous sodium sulfate and concentrated. The residue was purified by silica gel chromatography (eluent: 0-30% EtOAc/hexanes) to give 3,6-dichloro-5-(2-fluorophenyl)pyrazine-2-carboxamide.

^1H NMR (400 MHz, DMSO- _d6 ) δ: ppm 8.32 (br s, 1H), 8.16 (br s, 1H), 7.64-7.77 (m, 2H), 7, 41-7.47 (m, 2H).

Mass/charge (ESI, +ve): 286.0 (M+H) ⁺ .

Step 4. 3,6-Dichloro-5-(2-fluorophenyl)-N-((2-isopropylphenyl)carbamoyl)pyrazine-2-carboxamide.

A mixture of 3,6-dichloro-5-(2-fluorophenyl)pyrazine-2-carboxamide (704 mg, 2.46 mmol) and a solution of oxalyl chloride (2 M in DCM, 1.35 ml, 2.71 mmol) in DCE ( 15 ml) was heated at 80°C for 1 hour. The reaction mixture was cooled to room temperature and 2-isopropylaniline (0.35 ml, 2.46 mmol) was added. The resulting mixture was stirred at room temperature for 3 hours and then concentrated. The residue was treated with EtOAc, sonicated and filtered. The filtered solids were collected to give 3,6-dichloro-5-(2-fluorophenyl)-N-((2-155 044387 isopropylphenyl)carbamoyl)pyrazine-2-carboxamide.

Mass/charge (ESI, +ve): 446.8 (M+H)+.

Step 5. 6-Chloro-7-(2-fluorophenyl)-1-(2-isopropylphenyl)pteridine-2,4(III,3H)-dione (intermediate Y).

KHMDS (1 M solution in THF, 0.93 mL, 0.93 mmol) was added to the ice-cooled mixture of 3,6-dichloro-5-(2-fluorophenyl)-X-((2-isopropylphenyl)carbamoyl)pyrazine-2 -carboxamide (208 mg, 0.46 mmol) in THF (4 ml). The resulting mixture was allowed to warm to room temperature and stirred for 2 hours. The reaction mixture was partitioned between EtOAc (10 ml) and saturated aqueous ammonium chloride (2x5 ml). The organic layer was dried over anhydrous sodium sulfate and concentrated. The residue was purified by silica gel chromatography (eluent: 0-50% EtOAc/heptane) to give 6-chloro-7-(2-fluorophenyl)-1-(2-isopropylphenyl)pteridine-2,4(1H,3H)-dione .

¹ H NMR (400 MHz, CHLOROFORM-d) δ: ppm 8.50 (br s, 1H), 7.40-7.47 (m, 3H), 7.31 (dt, J=8.3, 4 ,2 Hz, 1H), 7.17-7.24 (m, 2H), 7.09-7.16 (m, 2H), 2.69 (quin, J=6.8 Hz, 1H), 1 .22 (d, J=6.8 Hz, 3H), 1.06 (d, J=6.8 Hz, 3H).

Mass/charge (ESI, +ve): 410.9 (M+H)+.

Step 6. 6-Chloro-7-(2-fluorophenyl)-4-((2S)-2-methyl-4-(2-propenoyl)-1-piperazinyl)-1-(2-(2propanyl)phenyl)- 2( 1 H)-pteridinone.

DIPEA was added to a solution of 6-chloro-7-(2-fluorophenyl)-1-(2-isonropylphenyl)pteridine-2,4(1H,3H)-dione (373 mg, 0.91 mmol) in acetonitrile (5 ml). (0.269 ml, 1.54 mmol) followed by the addition of phosphorus oxychloride (0.127 ml, 1.36 mmol). The resulting solution was stirred at 80°C for 30 minutes and then concentrated. The residue was dissolved in DMF (5 ml), cooled to 0°C and treated with DIPEA (0.54 ml, 3.08 mmol), then with a solution of (S)-1-(3-methylpiperazin-1-yl) prop-2en-1-one-2,2,2-trifluoroacetate (582 mg, 1.00 mmol) in DMF (0.5 ml). The resulting solution was stirred at 0°C for 30 minutes, and then gradually warmed to room temperature. The reaction mixture was partitioned between EtOAc (10 ml) and water (2x10 ml). The organic layer was dried over anhydrous sodium sulfate and concentrated. The residue was purified by silica gel chromatography (eluent: 0-3% MeOH/DCM) to give 6-chloro-7-(2-fluorophenyl)-4-((2S)-2-methyl-4-(2propenoyl)-1 - piperazinyl)-1 -(2-(2-propanyl)phenyl)-2(1H)-pterudinone.

^1H NMR (400 MHz, DMSO-d6) δ: ppm 7.51-7.61 (m, 1H), 7.44 (dt, J=7.9, 1.9 Hz, 1H), 7.33 -7.38 (m, 2H), 7.27-7.32 (m, 2H), 7.24 (br t, J=7.7 Hz, 1H), 7.13 (br dt, J=7 ,9, 1.7 Hz, 1H), 6.89 (br dd, J=16.4, 10.4 Hz, 1H), 6.22 (br d, J=16.4 Hz, 1H), 5 .76 (dd, J=10.2, 1.2 Hz, 1H), 4.68-5.54 (m, 1H), 4.21-4.44 (m, 6H), 2.69-2 .83 (m, 1H), 1.21-1.31 (m, 3H), 1.10 (t, J=6.3 Hz, 3H), 0.99 (dd, J=6.7, 3 ,4 Hz, 3H).

Mass/charge (ESI, +ve): 546.8 (M+H)+.

Example 51.

7-(2-Fluorophenyl)-6-methyl-4-((2S)-2-methyl-4-(2-πpropenoyl)-1-πiperazinyl)-1-(2-(2-propanyl)phenyl)-2 (1H)-pteridinone

Step 1. 7-(2-fluorophenyl)-1-(2-isopropylphenyl)-6-methylpteridine-2,4(1H,3H)-dione.

A mixture of 6-chloro-7-(2-fluorophenyl)-1-(2uzopropylphenyl)pteridine-2,4(1H,3H)-dione (intermediate Y, 930 mg, 2.27 mmol) was charged into a glass microwave reaction vessel. [1,1'-bis-(diphenylphosphino)ferrocene]dichloropalladium(II) complexed with dichloromethane (166 mg, 0.28 mmol) trimethylboroxine (0.64 ml, 4.54 mmol, Sigma-Aldrich, St. Louis, Missouri, USA) and potassium carbonate (0.63 g, 4.54 mmol) in 10:1 dioxane/water (11 ml). The reaction mixture was degassed with nitrogen for 5 min and heated in an Emrys Optimizer microwave reactor (Biotage, Uppsala, Sweden) at 100°C for 1.5 h. The reaction mixture was allowed to cool to room temperature and then partitioned between water ( 10 ml) and EtOAc (10 ml). The organic layer was dried over anhydrous sodium sulfate and concentrated. The residue was purified by silica gel chromatography (eluent: 0-40% EtOAc/heptane) to give 7-(2-fluorophenyl)-1-(2-isopropylphenyl)-6methylπterudin-2,4(1H,3H)-dione.

¹ H NMR (400 MHz, CHLOROFORM-d) δ: ppm 8.53 (br s, 1H), 7.38-7.48 (m, 3H), 7.30 (dt, J=8.3, 4 ,2 Hz, 1H), 7.08-7.20 (m, 4H), 2.71 (quin, J=6.8 Hz, 1H), 2.61 (d, J=2.3 Hz, 3H ), 1.21 (d, J=6.8 Hz, 3H), 1.05 (d, J=6.8 Hz, 3H).

Mass/charge (ESI, +ve): 391.0 (M+H)+.

- 156 044387

Step 2. 7-(2-Fluorophenyl)-6-methyl-4-((2S)-2-methyl-4-(2-propenoyl)-1-piperazinyl)-1-(2-(2propanyl)phenyl)- 2(1H)-pteridinone.

DIPEA ( 0.252 ml, 1.44 mmol) followed by the addition of phosphorus oxychloride (0.119 ml, 1.27 mmol). The resulting solution was stirred at 60°C for 1 hour and then concentrated. The residue was dissolved in DMF (5 ml), cooled to 0°C and treated with DIPEA (0.51 ml, 2.88 mmol), then with a solution of ^)-1-(3-methylpiperazin-1-yl)prop -2en-1-one-2,2,2-trifluoroacetate (544 mg, 0.93 mmol) in DMF (0.5 ml) using a cannula. The resulting mixture was stirred at 0°C for 30 minutes and then gradually warmed to room temperature. The reaction mixture was partitioned between EtOAc (10 ml) and water (2x10 ml). The organic layer was dried over anhydrous sodium sulfate and concentrated. The residue was purified by silica gel chromatography (eluent: 0-3% MeOH/DCM) to give 7-(2-fluorophenyl)-6-methyl4-((2S)-2-methyl-4-(2-propenoyl)-1 - piperazinyl)-1-(2-(2-propanyl)phenyl)-2(1H)-pteridinone.

Ή NMR (400 MHz, DMSO-d6) δ: ppm 7.48-7.57 (m, 1H), 7.42 (dt, J=7.7, 1.9 Hz, 1H), 7.32- 7.37 (m, 2H), 7.20-7.31 (m, 3H), 7.10 (br dt, J=7.1, 1.0 Hz, 1H), 6.80-6.95 (m, 1H), 6.20 (br d, J=16.6 Hz, 1H), 5.75 (dd, J=10.3, 2.0 Hz, 1H), 4.66-5.57 (m, 1H), 4.13-4.46 (m, 3H), 3.44-3.73 (m, 3H), 2.56-2.70 (m, 1H), 2.42 (s , 3H), 1.25 (br s, 3H), 1.09 (t, J=6.1 Hz, 3H), 0.97 (dd, J=6.6, 1.7 Hz, 3H).

Mass/charge (ESI, +ve): 527.0 (M+H) ⁺ .

Example 52 7-(2-Fluoro-6-hydroxyphenyl)-6-methyl-4-((2S)-2-methyl-4-(2-propenoyl)-1-piperazinyl)1-(2-(2-propanyl) )phenyl)pyrido[2,3-d]pyrimidin-2(1H)-one

Stage 1. 2,6-Dichloro-5-methylnicotinic acid.

A mixture of ethyl 2,6-dichloro-5-methylnicotinate (6.49 g, 27.7 mmol, Pharmablock Inc., Sunnyvale, CA, USA) in TFA (30 ml) and 5 N. HCl (24 ml) was heated at 90°C for 16 hours. The reaction mixture was allowed to cool to room temperature and then partially concentrated. Water was added and the mixture was filtered. The filtered solids were collected and dried under vacuum to obtain 2,6-dichloro-5-methylnicotinic acid.

Mass/charge (ESI, +ve): 205.9 (M+H) ⁺ .

Stage 2. 2,6-Dichloro-5-methylnicotinamide.

To an ice-cooled mixture of 2,6-dichloro-5-methylnicotinic acid (4.55 g, 22.1 mmol) in DCM (30 mL) was added oxalyl chloride (2 M solution in DCM, 16.6 mL, 33.1 mmol) followed by adding a few drops of DMF. The reaction mixture was gradually warmed to room temperature and stirred for 1 hour and then concentrated. The residue was suspended in toluene (15 ml), cooled to 0°C and treated with ammonium hydroxide (30%, 9.1 ml, 62 mmol). The reaction mixture was stirred at room temperature for 30 minutes and then filtered. The filtered solids were washed with water and dried under vacuum to obtain 2,6-dichloro-5-methylnicotinamide.

Mass/charge (ESI, +ve): 204.9 (M+H) ⁺ .

Step 3. 2,6-Dichloro-N-((2-isopropylphenyl)carbamoyl)-5-methylnicotinamide.

A mixture of 2,6-dichloro-5-methylnicotinamide (513 mg, 2.50 mmol) and oxalyl chloride (2 M solution in DCM, 1.38 ml, 2.63 mmol) in THF (10 ml) was heated at 65°C in for 1 hour. The reaction mixture was cooled to room temperature and 2-isopropylaniline (0.36 ml, 2.63 mmol) was added. The resulting mixture was stirred at room temperature for 1 hour and then concentrated. The residue was partitioned between EtOAc (50 ml) and saturated aqueous sodium bicarbonate (15 ml). Organic

- 157 044387 the layer was dried over anhydrous sodium sulfate and concentrated. The residue was suspended in heptane/EtOAc 5:1 and filtered. The filtered solids were collected to give 2,6-dichloroK-((2-isopropylphenyl)carbamoyl)-5-methylnicotinamide.

Mass/charge (ESI, +ve): 365.8 (M+H) ⁺ .

Step 4. 7-Chloro-1-(2-isopropylphenyl)-6-methylpyrido[2,3-d]pyrimidine-2,4(1H,3H)-dione.

KHMDS (1 M solution in THF, 4.54 mL, 4.54 mmol) was added via syringe to an ice-cold mixture of 2,6-dichloro-K-((2-isopropylphenyl)carbamoyl)-5-methylnicotinamide (831 mg, 2.27 mmol) in THF (10 ml). The resulting mixture was allowed to warm to room temperature and stirred for 2 hours. The reaction mixture was partitioned between EtOAc (20 ml) and saturated aqueous ammonium chloride (2x10 ml). The organic layer was dried over anhydrous sodium sulfate and concentrated. The residue was suspended in heptane/EtOAc 5:1 and filtered. The filtered solids were collected to give 7-chloro-1-(2-isopropylphenyl)-6-methylpyrido[2,3-d]pyrimidine-2,4(1H,3H)-dione.

¹ H NMR (400 MHz, DMSO d ₆ ) δ: ppm 8.36 (s, 1H), 7.49 (dd, J=8.5, 1.4 Hz, 1H), 7.44 (td, J =6.8, 1.2 Hz, 1H), 7.26-7.33 (m, 1H), 7.23 (dd, J=7.9, 1.7 Hz, 1H), 2.68 ( quin, J=6.8 Hz, 1H), 2.34 (s, 3H), 1.08 (d, J=6.8 Hz, 3H), 1.02 (d, J=6.6 Hz, 3H).

Mass/charge (ESI, +ve): 329.9 (M+H) ⁺ .

Step 5. tert-Butyl-(S)-4-(7-chloro-1-(2-isopropylphenyl)-6-methyl-2-oxo-1,2-dihydropyrido[2,3b]pyrimidin-4-yl) -3-methylpiperazine-1-carboxylate.

To a solution of 7-chloro-1-(2-isopropylphenyl)-6-methylpyrido[2,3-b]pyrimidine-2,4(1H,3H)-dione (444 mg, 1.35 mmol) in acetonitrile (5 ml ) DIPEA (0.70 mL, 4.04 mmol) was added followed by phosphorus oxychloride (0.125 mL, 1.35 mmol). The resulting solution was stirred at 80°C for 2 hours and then concentrated. The residue was dissolved in acetonitrile (5 ml), cooled to 0°C and treated with DIPEA (0.70 ml, 4.04 mmol), then with (S)-4-N-Boc-2methylpiperazine (297 mg, 1 .48 mmol, Combi-Blocks, Inc., San Diego, CA, USA). The resulting mixture was stirred at 0°C for 30 minutes and then gradually warmed to room temperature. The reaction mixture was partitioned between EtOAc (10 ml) and water (2x10 ml). The organic layer was dried over anhydrous sodium sulfate and concentrated. The residue was purified by silica gel chromatography (eluent: 0-30% 3:1 EtOAc-EtOH/heptane) to give tert-butyl(S)-4-(7-chloro-1-(2-isopropylphenyl)-6-methyl- 2-oxo-1,2-dihydropyrido[2,3-b]pyrimidin-4-yl)-3methylpiperazine-1-carboxylate.

1H NMR (400 MHz, DMSO-d6) δ: ppm 8.16 (d, J=16.2 Hz, 1H), 7.47 (dd, J=6.8, 1.2 Hz, 1H), 7 .41 (br t, J=7.3 Hz, 1H), 7.28 (dt, J=7.7, 1.2 Hz, 1H), 7.09 (dt, J=7.8, 1, 6 Hz, 1H), 4.70-4.90 (m, 1H), 3.89-4.20 (m, 2H), 3.97-4.09 (m, 2H), 3.76-3 .88 (m, 1H), 3.53-3.72 (m, 1H), 2.44 (td, J=6.8, 4.9 Hz, 1H), 2.35 (d, J=0 .8 Hz, 3H), 1.45 (s, 9H), 1.25 (br s, 3H), 1.06 (d, J=6.8, 3H), 1.02 (dd, J=6 ,8, 1.7 Hz, 3H).

Mass/charge (ESI, +ve): 511.9 (M+H) ⁺ .

Step 6. tert-Butyl-(3 S)-4-(7-(2-fluoro-6-hydroxyphenyl)-1 -(2-isopropylphenyl)-6-methyl-2-oxo1,2-dihydropyrido[2,3 -d]pyrimidin-4-yl)-3-methylpiperazine-1-carboxylate.

Mixture of tert-butyl-(S)-4-(7-chloro-1-(2-isopropylphenyl)-6-methyl-2-oxo-1,2-dihydropyrido[2,3d]pyrimidin-4-yl)-3 -methylpiperazine-1-carboxylate (458 mg, 0.89 mmol), potassium acetate (439 mg, 4.47 mmol), (2-fluoro-6-hydroxyphenyl)boronic acid (418 mg, 2.68 mmol, Combi- Blocks, San Diego, CA, USA) and [1,1'-bis(diphenylphosphino)ferrocene]-dichloropalladium(I) complexed with dichloromethane (65 mg, 0.09 mmol) in 1,4-dioxane (7 ml) and water (0.05 was heated at 90°C for 2 hours. Allow the reaction mixture to cool to room temperature and then partition it between saturated aqueous sodium bicarbonate (15 ml) and EtOAc (20 ml). The organic layer was washed with saline solution (10 ml), dried over anhydrous sodium sulfate and concentrated.The residue was purified by chromatography on silica gel (eluent: 0-30% 3:1 EtOAc-EtOH/heptane) to give tert-butyl-(3S)-4-( 7-(2-fluoro-6-hydroxyphenyl)-1-(2-isopropylphenyl)-6-methyl-2-oxo-1,2-dihydropyrido[2,3-d]pyrimidin-4-yl)-3-methylpiperazine -1-carboxylate.

Mass/charge (ESI, +ve): 587.9 (M+H) ⁺ .

Step 7. 7-(2-Fluoro-6-hydroxyphenyl)-6-methyl-4-((2S)-2-methyl-4-(2-propenoyl)-1-piperazinyl)-1(2-(2- propanyl)phenyl)pyrido[2,3-d]pyrimidin-2(1H)-one.

Trifluoroacetic acid (1.7 mL, 14.7 mmol) was added to the solution of tert-butyl-(3S)-4-(7-(2-fluoro6-hydroxyphenyl)-1-(2-isopropylphenyl)-6-methyl-2 -oxo-1,2-dihydropyrido[2,3-d]pyrimidin-4-yl)-3methylpiperazine-1-carboxylate (288 mg, 0.49 mmol) in DCM (5 ml). The resulting mixture was stirred at room temperature for 30 minutes and then concentrated. The residue was dissolved in DCM (3 ml), cooled to 0°C and treated with DIPEA (0.34 ml, 1.96 mmol), then with a solution of acryloyl chloride (0.042 ml, 0.51 mmol) in DCM (0. 5 ml). The reaction mixture was warmed to room temperature and stirred for 30 min, then quenched with saturated aqueous sodium bicarbonate (5 ml) and extracted with EtOAc (10 ml). The organic layer was washed with brine (5 ml), dried over anhydrous sodium sulfate and concentrated. The residue was purified by chromatography on silica gel (eluent: 0-50% 3:1 EtOAc-EtOH/heptane) with

- 158 044387 by preparing 7-(2-fluoro-6-hydroxyphenyl)-6-methyl-1-(2-methyl-6-(2-propanyl)phenyl)-4-((2S)-2-methyl-4- (2propenoyl)-1-piperazinyl)pyrido[2,3D]pyrimidin-2(III)-one.

Ή NMR (400 MHz, DMSO- _d6 ) δ: ppm 9.94 (s, 1H), 8.10-8.19 (m, 1H), 7.38 (dd, J=7.5, 1.0 Hz, 1H), 7.31 (td, J=7.1, 0.8 Hz, 1H), 7.16-7.24 (m, 2H), 7.06 (br dt, J=7.9 , 1.9 Hz, 1H), 6.75-6.97 (m, 1H), 6.71 (d, J=8.1 Hz, 1H), 6.64 (t, J=8.8 Hz , 1H), 6.21 (br dd, J=16.9, 5.7 Hz, 1H), 5.76 (dd, J=10.2, 2.5 Hz, 1H), 4.76-4 .98 (m, 1H), 4.10-4.51 (m, 2H), 3.24-3.81 (m, 4H), 2.43-2.49 (m, 1H), 2.12 (d, J=2.9 Hz, 3H), 1.22-1.33 (m, 3H), 1.05 (d, J=6.8 Hz, 3H), 0.96 (d, J= 6.8 Hz, 3H).

Mass/charge (ESI, +ve): 541.8 (M+H) ⁺ .

Example 53.

6-Chloro-7-(2-fluorophenyl)-1-(4-methyl-1-(2-nponanyl)-1H-nirazol-5-yl)-4-((2S)-2-methyl-4-( 2propenoyl)-1-piperazinyl)pyrido[2,3-d]pyrimidin-2(1H)-one (1) (C0CI) ₂ , THF, 65 °C (2)

connection P

Step 1. 2,5,6-Trichloro-N-((1-isopronyl-4-methyl-1H-pyrazol-5-yl)carbamoyl)nicotinamide.

To a mixture of 2,5,6-trichloronicotinamide (intermediate P, 2.43 g, 10.8 mmol) in THF (21.5 mL) was added a solution of oxalyl chloride (2 M in DCM, 5.70 mL, 11.4 mmol ) at room temperature. The resulting mixture was heated at 65°C for 3 hours, then the heating was stopped and the reaction mixture was cooled to 0°C. A solution of 1-isopropyl-4-methyl-1H-pyrazol-5amine (1.50 g, 10.8 mmol, ChemBridge, San Diego, CA, USA) in THF (15 ml) was added via cannula. The resulting mixture was stirred at 0°C for 30 minutes and then warmed to room temperature and stirred for an additional 16 hours. The reaction mixture was filtered and the filtered solids were collected and washed with acetonitrile (20 ml) to obtain 2,5,6trichloro -N-(( 1 -isopropyl-4-methyl-1 H-pyrazol-5 -yl)carbamoyl)nicotinamide.

^1H NMR (400 MHz, DMSO-d6) δ: ppm 11.39 (br s, 1H), 9.51 (br s, 1H), 8.62 (s, 1H), 7.29 (s, 1H ), 4.334.44 (m, 1H), 1.87 (s, 3H), 1.33 (d, J=6.6 Hz, 6H).

Mass/charge (ESI, +ve ion): 389.9 (M+H) ⁺ .

Step 2. 6,7-Dichloro-1-(1-isopropyl-4-methyl-1H-pyrazol-5-yl)pyrido[2,3-d]pyrimidin-

2,4(1H,3H)-dione.

KHMDS (1 M solution in THF, 18.0 mL, 18.0 mmol) was added slowly via syringe to the ice-cold mixture of 2,5,6-trichloro-N-((1-isopropyl-4-methyl-1H-pyrazole -5-yl)carbamoyl)nicotinamide (3.52 g, 9.0 mmol) in THF (30 ml). After 10 minutes, the resulting mixture was allowed to warm to room temperature and stirred for 18 hours. The resulting slurry was quenched with saturated aqueous ammonium chloride (50 ml) and brine (50 ml), and then extracted with EtOAc (2x50 ml). The combined organic layers were washed with brine (50 ml), dried over anhydrous sodium sulfate and concentrated to give 6,7-dichloro-1-(1isopropyl-4-methyl-1H-pyrazol-5-yl)pyrido[2,3 -d]nirimidine-2,4(1H,3H)-dione. This material was used in the next step without further purification.

1H NMR (400 MHz, DMSO- _d6 ) δ: ppm 12.22 (br s, 1H), 8.55 (s, 1H), 7.45 (s, 1H), 4.22-4.34 ( m, 1H), 1.78 (s, 3H), 1.25 (t, J=6.6 Hz, 6H).

Mass/charge (ESI, +ve ion): 353.9 (M+H) ⁺ .

- 159 044387

Step 3. 4,6,7-Trichloro-1-(1-isopropyl-4-methyl-1H-pyrazol-5-yl)pyrido[2,3-d]pyrimidin-2(1H)-one.

To a solution of 6,7-dichloro-1-(1-isopropyl-4-methyl-1H-pyrazol-5-yl)pyrido[2,3-d]pyrimidine2,4(1H,3H)-dione (3.19 g, 9.0 mmol) and DIPEA (4.71 mL, 27.0 mmol) in acetonitrile (60 mL), phosphorus oxychloride (1.68 mL, 18.0 mmol) was added. The resulting mixture was heated at 80°C for 30 minutes and then cooled to room temperature and concentrated to give 4,6,7-trichloro-1-(1isopropyl-4-methyl-1H-pyrazol-5-yl)pyrido[2 ,3-d]pyrimidin-2(1H)-one. This material was used in the next step without further purification.

Step 4. (S)-tert-Butyl-4-(6,7-dichloro-1 -(1-isopropyl-4-methyl-1H-pyrazol-5-yl)-2-oxo-1,2dihydropyrido[2 ,3-d]pyrimidin-4-yl)-3-methylpiperazine-1-carboxylate.

To a solution of 4,6,7-trichloro-1-(1-isopropyl-4-methyl-1H-pyrazol-5-yl)pyrido[2,3-d]pyrimidin-2(1H)one (3.36 g , 9.0 mmol) in THF (45 ml) was added DIPEA (4.71 ml, 27.1 mmol) followed by the addition of (S)-4-N-Boc-2-methylpiperazine (2.71 g, 13.6 mmol, Combi-Blocks, Inc., San Diego, CA, USA). The resulting mixture was stirred at room temperature for 1 hour, then quenched with ice-cold saturated aqueous sodium bicarbonate (100 ml) and extracted with EtOAc (100 ml, 2x50 ml). The combined organic layers were dried over anhydrous sodium sulfate and concentrated. The residue was purified by silica gel chromatography (eluent: 0-50% 3:1 EtOAc-EtOH/heptane) to give (S)-tert-butyl-4-(6,7-dichloro-1-(1-isopropyl-4methyl- 1H-pyrazol-5-yl)-2-oxo-1,2-dihydropyrido[2,3-d]pyrimidin-4-yl)-3-methylpiperazine-1carboxylate.

1H NMR (400 MHz, DMSO-d6) δ: ppm 8.43 (d, J=19.1 Hz, 1H), 7.42 (s, 1H), 4.77-4.98 (m, 1H) , 4.15 (br dd, J=26.7, 13.5 Hz, 1H), 3.61-4.07 (m, 4H), 2.94-3.28 (m, 2H), 1, 72 (d, J=5.2 Hz, 3H), 1.44 (s, 9H), 1.32 (br dd, J=13.0, 6.7 Hz, 3H), 1.21-1, 28 (m, 6H).

Mass/charge (ESI, +ve ion): 536.0 (M+H) ⁺ .

Step 5. tert-Butyl-(S)-4-(6-chloro-7-(2-fluorophenyl)-1-(1-isopropyl-4-methyl-1H-pyrazol-5-yl)-2oxo-1, 2-dihydropyrido[2,3-d]pyrimidin-4-yl)-3-methylpiperazine-1-carboxylate.

A mixture of tert-butyl-(S)-4-(6,7-dichloro-1-(1-isopropyl-4-methyl-1H-pyrazol-5-yl)-2-oxo-1,2-dihydropyrido[2, 3-d]pyrimidin-4-yl)-3-methylpiperazine-1-carboxylate (703 mg, 1.31 mmol), (2-fluorophenyl)boronic acid (330 mg, 2.36 mmol, Combi-Blocks, San- Diego, California, USA), anhydrous sodium carbonate (416 mg, 3.93 mmol) and tetrakis(triphenylphosphine)palladium(0) (151 mg, 0.13 mmol) in 1,4-dioxane (3.5 ml) and water (0.87 ml) were heated at 80°C for 2 hours. The reaction mixture was concentrated and the residue was purified by silica gel chromatography (eluent: 0-100% EtOAc/heptane) to give tert-butyl-(S)-4 -(6-chloro-7-(2-fluorophenyl)-1-(1-isopropyl-4-methyl-1H-pyrazol-5-yl)-2-oxo1,2-dihydropyrido[2,3-d]pyrimidin- 4-yl)-3-methylpiperazine-1-carboxylate.

1H NMR (400 MHz, DMSO-d6) δ: ppm 8.41 (d, J=8.9 Hz, 1H), 7.50-7.66 (m, 1H), 7.24-7.37 ( m, 4H), 4.92 (br s, 1H), 4.24 (br t, J=12.4 Hz, 1H), 3.95-4.12 (m, 2H), 3.67-3 .89 (m, 2H), 3.00-3.29 (m, 2H), 1.70 (s, 3H), 1.45 (s, 9H), 1.36 (t, J=6.6 Hz, 3H), 1.14-1.28 (m, 6H).

¹⁹ F NMR (376 MHz, DMSO-d ₆ ) δ: ppm -113.95 (d, J=6.9 Hz, 1F).

Mass/charge (ESI, +ve ion): 596.0 (M+H) ⁺ .

Step 6. 6-Chloro-7-(2-fluorophenyl)-1-(4-methyl-1-(2-propanyl)-1H-pyrazol-5-yl)-4-((2S)-2-methyl- 4(2-propenoyl)-1-piperazinyl)pyrido[2,3-d]pyrimidin-2(1H)-one.

Trifluoroacetic acid (7.4 mL, 63.6 mmol) was added to a solution of tert-butyl-(S)-4-(6-chloro-7-(2fluorophenyl)-1-(1-isopropyl-4-methyl-1H- pyrazol-5-yl)-2-oxo-1,2-dihydropyrido[2,3-b]pyrimidin-4yl)-3-methylpiperazine-1-carboxylate (532 mg, 0.74 mmol) in DCM (7.4 ml). The resulting mixture was stirred at room temperature for 1 hour and then concentrated. The residue was dissolved in DCM (7.4 ml), cooled to 0°C and treated with DIPEA (1.28 ml, 7.35 mmol), then with acryloyl chloride (0.2 M solution in DCM, 3.67 ml , 0.74 mmol). The resulting mixture was stirred at 0°C for 20 min, then quenched with saturated aqueous sodium bicarbonate (50 ml) and extracted with DCM (2x50 ml). The combined organic layers were dried over anhydrous sodium sulfate and concentrated. The residue was purified by silica gel chromatography (eluent: 0-50% EtOAc/heptane, then 0-50% 4:1 DCM-MeOH/DCM) to give 6-chloro-7-(2-fluorophenyl)-1-(4methyl- 1-(2-propanyl)-1H-pyrazol-5-yl)-4-((25)-2-methyl-4-(2-propenoyl)-1-piperazinyl)pyrido[2,3d]pyrimidine-2( 1 N)-she.

1H NMR (400 MHz, DMSO-d ₆ ) δ: ppm 8.44 (br d, J=7.5 Hz, 1H), 7.50-7.59 (m, 1H), 7.23-7, 41 (m, 4H), 6.77-6.93 (m, 1H), 6.21 (br d, J=16.4 Hz, 1H), 5.71-5.81 (m, 1H), 4.96 (br s, 1H), 4.22-4.46 (m, 2H), 3.96-4.21 (m, 2H), 3.37-3.89 (m, 2H), 3 ,00-3.28 (m, 1H), 1.71 (d, J=2.5 Hz, 3H), 1.34 (t, J=6.8 Hz, 3H), 1.12-1, 29 (m, 6H).

¹⁹ F NMR (376 MHz, DMSO-d ₆ ) δ: ppm -113.96 (d, J=9.5 Hz, 1F).

Mass/charge (ESI, +ve ion): 550.2 (M+H) ⁺ .

- 160 044387

Table 12(a)

Examples of separated compounds, including stereoisomers, some of which are atropoisomers

No. note Chemical structure Name Racemic SM/Separation Conditions 2-5-1 o "A A ^S N- ^J N h °" μ J Г 1 1st eluting isomer 1-((35)-4-(5-chloro-6(3-hydroxy-1naphthalenyl)[1,2]thiazol o[3,4-th]pyridin-3yl)-3-methyl-1piperazinyl)-2propen- 1-one or 1((37?)-4-(5-chloro-6-(3hydroxy-1naphthalenyl)[1,2]thiazol o[3,4-th]pyridin-3l)-3-methyl l- 1 piperazinyl)-2propen-1-one 2-5/ SFC (Chiralpak ADH, 20x150 mm, 5 µm, 55% MeOH/CO ₂ , 80 ml/min., 100 bar). 2-5-2 S P A | ' Ν ¹ А ОН .1 1 ( 1 2nd eluting isomer 1 -((37?)-4-(5-chloro-6(3-hydroxy-1naphthalenyl) [1,2]thiazol o[3,4-th]pyridin-3i l )-3-methyl l-1 piperazinyl )-2propen-1-one or 1((35)-4-(5-chloro-6-(3hydroxy-1naphthalenyl)[1,2]thiazol o[3,4-th]pyridin-3yl)-3-methyl -1piperazinyl)-2propen-1-one 2-5/ SFC (Chiralpak ADH, 20x150 mm, 5 µm, 55% MeOH/CO ₂ , 80 ml/min., 100 bar).

- 161 044387

2-6-1 V. ν Ν-, \ \ Ν μ-Α J ' ΜΗ ^Ν l· Ί ₁ „ ' ~ 1st eluting isomer 1-((35)-4-(5-chloro-6(5-methyl-1 H-ind azol4-yl)[l,2]τthiazolo[3,4L]pyridin-3-yl)-3methyl-1piperazinyl) -2propen-1-one or 1((37?)-4-(5-chloro-6-(5methyl-1 H-ind azol-4yl)[1,2]thiazolo[3,4L]pyridin-3-yl )-3methyl-1piperazinyl)-2propen-1-one 2-6/ SFC (Phenomenex (S, S)-Whelk-0 1, 250x20 mm, 3 µm, 50% MeOH/CO ₂ containing 20 mM NH ₃ , 60 g/min., 102 bar) 2-6-2 Έ ο Ν·^ λ-Ν ' Υ - ^α _{Λ c} - r 4 _K| J - ί NH ^Ν Ί ί ₉ „ - - 2nd eluting isomer 1 -((37?)-4-(5-chloro-6(5-methyl-1 H-ind azol4-yl)[1,2]thiazolo[3,4- L]pyridin-3-yl)-3methyl-1piperazinyl)-2propen-1-one or 1((35)-4-(5-chloro-6-(5methyl-1H-indazol-4yl)[1,2 ]thiazolo[3,4L]pyridin-3-yl)-3methyl-1piperazinyl)-2propen-1-one 2-6/ SFC (Phenomenex (S, S)-Whelk-0 1, 250x20 mm, 3 µm, 50% MeOH/CO ₂ containing 20 mM NH ₃ , 60 g/min., 102 bar)

- 162 044387

1-19-1 o.- _h Ll V'l II g 1 1 JI OH 1st eluting isomer 1-((3 8)-4-(5-chloro-7fluoro-6-(3-hydroxy1-naphthalenyl)-2,1benzothiazol-3-yl)-3methyl-1piperazinyl)-2- propen-1-one 1-19/ SFC (IC 250x30 mm, 5 µm, 50% MeOH/CO ₂ (w/ 20 mM NH ₃ ), 100 g/min., 100 bar). 1-19-2 O D. C| % 1 ^! . ¹ L A 2nd eluting isomer 1-((3 8)-4-(5-chloro-7fluoro-6-(3-hydroxy1-naphthalenyl)-2,1benzothiazol-3-yl)-3methyl-1piperazinyl)-2- propen-1-one 1-19/ SFC (IC 250x30 mm, 5 µm, 50% MeOH/CO ₂ (w/ 20 mM NH ₃ ), 100 g/min., 100 bar). 3-1-1 h° Oh I uu 1st eluting isomer 1 -(4-(5-chloro-7-fluoro6-(3-hydroxy-1naphthalenyl)-2,1benzothiazol-3-yl)-1 piperazinyl)-2propen-1-one 3-1/ SFC (OD-H 250x21 mm, 5 µm 40% MeOH/CO ₂ (w/ 20 mM NH ₃ ), 60 ml/min., 100 bar). 3-1-2 U Q l .He 2nd eluting isomer 1 -(4-(5-chloro-7-fluoro6-(3-hydroxy-1naphthalenyl)-2,1benzothiazol-3-yl)-1 piperazinyl)-2propen-1-one 3-1/ SFC (OD-H 250x21 mm, 5 µm 40% MeOH/CO ₂ (w/ 20 mM NH ₃ ), 60 ml/min., 100 bar).

- 163 044387

8-6-1 V/ί U N 0 ’ Ν Ν~ γ > 1 ί but'4‘ ' V 1st eluted isomer 6-chloro-7-(2-fluoro-6hydroxyphenyl)-4((2 S)-2-methyl-4-(2πpoπenoyl)-lpiperazinyl)-1 -(2-(2propanyl)phenyl)pyride o[2,3 -s!]pyrimidine- 2(1H)-on 8-6/ SFC (Chiralpak IC, 150x30 mm, 5 µm, 30% MeOH/CO ₂ (w/20 mM NH ₃ ), 120 g/min, 102 bar). 8-6-2 at /V Agricultural 2nd eluted isomer 6-chloro-7-(2-fluoro-6hydroxyphenyl)-4((2 S)-2-methyl-4-(2πpoπenoyl)-lpiperazinyl)-1 -(2-(2propanyl)phenyl)pyride o[2,3 -s!]pyrimidine- 2(1H)-on 8-6/ SFC (Chiralpak IC, 150x30 mm, 5 µm, 30% MeOH/CO ₂ (w/20 mM NH ₃ ), 120 g/min, 102 bar). 8-1-1 U eleven M' - <-1, G 3 Ί [ 1 | Ί J 1 ί but 1! 1 1st eluted isomer 6-chloro-7-(2-fluoro-6hydroxyphenyl)-1 -(2(2-propanyl)phenyl)4-(4-(2-propenoyl)-1 piperazinyl)-2(1H)- quinazolinone 8-1/ SFC (Chiralpak IC, 300x15 mm, 5 µm, 40% MeOH/CO ₂ (w/20 mM NH ₃ ), 135 g/min, 188 bar). 8-1-2 V N. : ] 'Ν g e Ί Ί Ί A y i 6-chloro-7-(2-fluoro-6hydroxyphenyl)-1 -(2(2-propanyl)phenyl)4-(4-(2-propenoyl)-1 piperazinyl)-2(1H)- quinazolinone 8-1/ SFC (Chiralpak IC, 300x15 mm, 5 µm, 40% MeOH/CO ₂ (w/ 20mMNH ₃ ), 135 r/min., 188 bar).

- 164 044387

2nd eluting isomer U a a 6-chloro-7-(2-fluoro-6- 8-3/ hydroxyphenyl)-4- SFC „X Xi _with ^N XA' ^h L JX ..1.. Ail ((28)-2-methyl-4-(2- (Whelk-01 (S, S), 8-3-1 propenoyl)-1- 250x21 mm, 5 microns, 1 BUT' 1st eluted piperazinyl)-1 -(2-(2- 30% EtOH/CO ₂ (w/ 20 propanyl)phenyl)2(1H)-quinazolinone mM NH ₃ ), 70 g/min, 187 bar). isomer J _ν O Ν 6-chloro-7-(2-fluoro-6- 8-3/ 1 . \ hydroxyphenyl)-4- SFC = CI, Ν η _; h LJ ί L ? J G ] ((2 8)-2-methyl-4-(2- (Whelk-01 (S, S), 8-3-2 propenoyl)-1- 250x21 mm, 5 microns, I: and<. ^L > g ^% piperazinyl)-1 -(2-(2- 30% EtOH/CO ₂ (w/ 20 II, propanyl)phenyl)- mM NH ₃ ), 70 g/min, 2nd eluted 2(1H)-quinazolinone 187 bar). Example isomer s separated connections opinions, including Table 12(b) of tropoisomers Note No. Chemical structure Name Racemic SM/conditions 39-1 V/ N—. WITH? F Ν —N <— 6-chloro-7-(2-fluoro-6hydroxyphenyl)-1 -(2methyl-6-(2- separation 39/ SFC (Chiralpak ΟΠ- Η, 21x250 mm, 5 µm, >—N BUT 0 1st eluting isomer propanyl)phenyl)-4((28)-2-methyl-4-(2propenoyl)-!- 30% MeOH/CO ₂ , 50 ml/min., 100 bar).

- 165 044387

piperazinyl)pyrido[2, 3-d]pyrimidin-2(1H)one 39-2 w Μ ' 1 F /_- N—N' HC About >—< 2nd eluting isomer 6-chloro-7-(2-fluoro-6hydroxyphenyl)-1 -(2methyl-6-(2propanyl)phenyl)-4((2 8)-2-methyl-4-(2propenoyl)-1- piperazinyl)pyrido[2, 3-d] pyrimidine-2(1H)- He 39/ SFC (Chiralpak ΟΠΗ, 21x250 mm, 5 µm, 30% MeOH/CO ₂ , 50 ml/min., 100 bar). 40-1 N—, N' Tn=' 1st eluting isomer 6-chloro-7-(2fluorophenyl)-1 -(4methyl-2-(2propanyl)-3pyridinyl)-4-((2 S)-2methyl-4-(2propenoyl)-1piperazinyl)pyrido[2, 3-d] pyrimidine-2(1H)- He 40/ SFC (Chiralpak IC, 21x150 mm, 5 µm, 60% MeOH/CO ₂ , 50 ml/min., 100 bar). 40-2 id 2nd eluting isomer 6-chloro-7-(2fluorophenyl)-1 -(4methyl-2-(2propanyl)-3pyridinyl)-4-((2 S)-2methyl-4-(2propenoyl)-1piperazinyl)pyrido[2, 3-d] pyrimidine-2(1H)- He 40/ SFC (Chiralpak IC, 21x150 mm, 5 µm, 60% MeOH/CO ₂ , 50 ml/min., 100 bar).

- 166 044387

Ch_ 6-fluoro-7-(2-fluoro-6hydroxyphenyl)-1 -(4methyl-2-(2- 41/ ί' ) F Г -Π propanyl)-3pyridinyl)-4-((2 S)-2- SFC (Chiralpak IC, 41-1 . He ο' \· <' Τ₽ 1st eluting isomer methyl-4-(2propenoyl)-1piperazinyl)pyrido[2, 3-d]pyrimidine-2(1H)- it is 6-fluoro-7-(2-fluoro-6hydroxyphenyl)-1 -(4- 30x250 mm, 5 µm, 55% MeOH/CO ₂ , 120 g/min., 103 bar). m Ν—, F F A, methyl-2-(2propanyl)-3pyridinyl)-4-((2S)-2- 41/SFC (Chiralpak IC, 41-2 N - N - сН O . η 2nd eluting isomer ψ O methyl-4-(2propenoyl)-1piperazinyl)pyrido[2, 3-d]pyrimidine-2(1H)- He 1 -(2-cyclopropyl-4methyl-3-pyridinyl)- 30x250 mm, 5 µm, 55% MeOH/CO ₂ , 120 g/min., 103 bar). Y-l 6-fluoro-7-(2-fluoro-6- 42/ η „-A hydroxyphenyl)-4- SFC (Chiralpak OD- 42-1 Ν g *- Ν PN ((2 S)-2-methyl-4-(2- H, 21x250 mm, 5 µm, WITH -. -l- 20% EUN/CO ₂ , 80 1st eluting isomer piperazinyl)pyrido[2, 3-d]pyrimidin-2(1H)one ml/min., 100 bar). 1 -(2-cyclopropyl-4methyl-3-pyridinyl)6-fluoro-7-(2-fluoro-6- 42/ SFC (Chiralpak ΟΠ- 42-2 Ν ί-Ν )—' A” 2nd eluting isomer hydroxyphenyl)-4((28)-2-methyl-4-(2propenoyl)-!- Η, 21x250 mm, 5 µm, 20% EUN/CO ₂ , 80 ml/min., 100 bar).

- 167 044387

piperazinyl)pyrido[2, 3-d]pyrimidin-2(1H)one 44-1 w N -> Ll F / n n :— UH - N— 1st eluting isomer 1 -(2-cyclopropyl-4methyl-3-pyridinyl)6-fluoro-7-(2-fluoro-6hydroxyphenyl)-4((2 S)-2-methyl-4-(2πpoπenoyl)-lpiperazinyl)pyrido[2, 3-d]pyrimidine-2(1H)- He 44/ SFC (Chiralpak ODN, 21x250 mm, 5 µm, 20% EUN/CO ₂ , 80 ml/min., 100 bar). 44-2 ft o N - / C ⁼ _ A tj Ί '— -N ΟΙ 1 O - N— 2nd eluting isomer 1 -(2-cyclopropyl-4methyl-3-pyridinyl)6-fluoro-7-(2-fluoro-6hydroxyphenyl)-4((2 S)-2-methyl-4-(2πpoπenoyl)-lpiperazinyl)pyrido[2, 3-d]pyrimidine-2(1H)- He 44/ SFC (Chiralpak ODH, 21x250 mm, 5 µm, 20% EtOH/CO ₂ , 80 ml/min., 100 bar). 45-1 vN,. t '> Cl F _s N y N ;> n . AH V _N =/ 1st eluting isomer 6-chloro-1-(2- cyclopropyl-4methyl-3-pyridinyl)7-(2-fluorophenyl)-4((2 S)-2-methyl-4-(2πpoπenoyl)-lpiperazinyl)pyrido[2, 3-d]pyrimidine-2(1H)- He 45/ SFC (Chiralpak ΟΠΗ, 21x250 mm, 5 µm, 20% EtOH/CO ₂ , 80 ml/min., 100 bar).

- 168044387

45-2 h N , ( \ Cl f lrk Pc. N y N -- AN _z °' pi V 2nd eluting isomer 6-chloro-1-(2cyclopropyl-4methyl-3-pyridinyl)7-(2-fluorophenyl)-4((2 S)-2-methyl-4-(2πpoπenoyl)-lpiperazinyl)pyrido[2, 3-d] pyrimidine-2(1H)- He 45/ SFC (Chiralpak ΟΠΗ, 21x250 mm, 5 µm, 20% EtOH/CO ₂ , 80 ml/min., 100 bar). 46-1 N—v ; Cl f U >—N ' —N (j '------ N ^is the 1st eluting isomer 6-chloro-1 -(2-ethyl-4methyl-3-pyridinyl)7-(2-fluorophenyl)-4((2 S)-2-methyl-4-(2πpoπenoyl)-lpiperazinyl)pyrido[2, 3- d]pyrimidine-2(1H)- He 46/ SFC (Chiralpak ADH, 20x250 mm, 5 µm, 30% i-PrOH/CO ₂ , 50 ml/min., 100 bar). 46-2 h Arb X / 2nd eluting isomer 6-chloro-1 -(2-ethyl-4methyl-3-pyridinyl)7-(2-fluorophenyl)-4((2 S)-2-methyl-4-(2πpoπenoyl)-lpiperazinyl)pyrido[2, 3- d]pyrimidine-2(1H)- He 46/ SFC (Chiralpak ADH, 20x250 mm, 5 µm, 30% i-PrOH/CO ₂ , 50 ml/min., 100 bar). 49-1 s, ϋ Ν— > Cl F y '· —' N—N ·— —P Hu U >...............l 1st eluting isomer 6-chloro-7-(2-fluoro-6hydroxyphenyl)-1 -(2(2-methyl-2propanyl)phenyl)-4((2 S)-2-methyl-4-(2πpoπenoyl)-lpiperazinyl)pyrido[2 , 3-d]pyrimidine-2(1H)- He 49/ SFC (Chiralpak IF, 21x250 mm, 5 µm, 50% MeOH/CO ₂ , 65 ml/min., 102 bar).

- 169 044387

49-2 1 < Cl p /' - N —fl — —N HC 2nd eluting isomer 6-chloro-7-(2-fluoro-6hydroxyphenyl)-1 -(2(2-methyl-2propanyl)phenyl)-4((2 S)-2-methyl-4-(2πpoπenoyl)-lpiperazinyl)pyrido[2 , 3-d]pyrimidine-2(1H)- He 49/ SFC (Chiralpak IF, 21x250 mm, 5 µm, 50% MeOH/CO ₂ , 65 ml/min., 102 bar). 50-1 η сАЪ 1st eluting isomer 6-chloro-7-(2fluorophenyl)-4-((28)-2methyl-4-(2propenoyl)-1piperazinyl)-1 -(2-(2propanyl)phenyl)2(1H)-pteridinone, 50/ SFC (Chiralpak OD, 21x250 mm, 5 µm, 17% MeOH/CO ₂ , 80 g/min., 102 bar). 50-2 ,l O N—\ (\af AAAA ν' An ' U ν' o' 2nd eluting isomer 6-chloro-7-(2fluorophenyl)-4-((28)-2methyl-4-(2propenoyl)-1piperazinyl)-1 -(2-(2propanyl)phenyl)2(1H)-pteridinone 50/ SFC (Chiralpak OD, 21x250 mm, 5 µm, 17% MeOH/CO ₂ , 80 g/min., 102 bar). 51-1 m°h Uch O u d -tUU 1st eluting isomer 7-(2-fluorophenyl)-6methyl-4-((28)-2methyl-4-(2propenoyl)-1piperazinyl)-1 -(2-(2propanyl)phenyl)2(1H)-pteridinone 51 / SFC ((S, S) Whelk01, 21x250 mm, 5 µm, 60% MeOH/CO ₂ , 60 g/min., 110 bar). 51-2 ' e Τ1Ύ ¹ k i t |/ k 7-(2-fluorophenyl)-6methyl-4-((28)-2methyl-4-(2propenoyl)-1piperazinyl)-1 -(2-(2- 51 / SFC ((S, S) Whelk01, 21x250 mm, 5 µm, 60% MeOH/CO ₂ , 60

- 170044387

2nd eluting isomer propanyl)phenyl)- 2(1H)-pteridinone g/min., 110 bar). 7-(2-fluoro-6- A O hydroxyphenyl)-6- A YagA ' „AT -N BUT methyl-4-((2S)-2- 52/ 52-1 methyl-4-(2- propenoyl)-1 - SFC (Chiralpak ASH, 21x250 mm, 5 µm, piperazinyl)-1 -(2-(2- 25% MeOH/CO ₂ , 50 1st eluting isomer propanyl)phenyl)pyride o[2,3-<1]pyrimidine- g/min., 125 bar). 2(1H)-on 7-(2-fluoro-6- \\ ° hydroxyphenyl)-6- methyl-4-((2S)-2- 52/ A/\Ta methyl-4-(2- SFC (Chiralpak AS- 52-2 ν Χν У Vn but tb propenoyl)-1piperazinyl)-1 -(2-(2- Η, 21x250 mm, 5 µm, 25% MeOH/CO ₂ , 50 2nd eluting isomer propanyl)phenyl)pyride o[2,3-<1]pyrimidine- g/min., 125 bar). 2(1H)-on 6-chloro-7-(2- fluorophenyl)-1 -(4- O methyl-1-(2- 53/ ..................A O A propanil)-1H- SFC((S,S)Whelk- 53-1 Vn A J pyrazol-5-yl)-4- ((28)-2-methyl-4-(2propenoyl)-1- 01, 21x250 mm, 5 µm, 25% MeOH/CO ₂ , 90 1st eluting isomer piperazinyl)pyrido[2, 3-d] pyrimidine-2(1H)- g/min., 102 bar). He 6-chloro-7-(2fluorophenyl)-1 -(4- A. U methyl-1-(2- 53/ About F propanil)-1H- SFC((S,S)Whelk- 53-2 / Ai ' J ¹ । ⁴ t 0 pyrazol-5-yl)-4((2 8)-2-methyl-4-(2- 01, 21x250 mm, 5 µm, 25% /N' propenoyl)-1- MeOH/CO ₂ , 90 2nd eluting isomer piperazinyl)pyrido[2, 3-d] pyrimidine-2(1H)- g/min., 102 bar). He

- 171 044387

Table 13

Analytical data for common procedures

Note No. LRMS: (ESI, +ve ion) mass/charge NMR 1-1 498.0 Ή NMR (400 MHz, DMSO-d ₆ ) δ 9.99 (br s, 1 Η), 8.04 (s, 1 Η), 7.55 (d, 7=8.7 Hz, 1 Η), 6.81-6.94 (m, 2 Η), 6.79 (d, 7=2.9 Hz, 1 Η), 6.19 (dd, 7=16.7, 2.2 Hz, 1 Η ), 5.77 (dd, 7=10.5, 2.2 Hz, 1 Η), 3.87 (br d, 7=19.5 Hz, 4 Η), 3.63 (br t, 7= 5.1 Hz, 4 H). ¹⁹ F NMR (376 MHz, DMSO-d ₆ ) δ 123.78 (s, 1 F). 1-2 441.1 Ή NMR (400 MHz, DMSO-7 ₆ ) δ 8.19 (s, 1H), 7.91 (d, 7=8.4 Hz, 1H), 7.45-7.53 (m, 2H), 7.43 (d, 7=2.4 Hz, 1H), 7.26-7.35 (m, 2H), 7.10 (d, 7=2.5 Hz, 1H), 6.87 (dd , 7=16.7, 10.5 Hz, 1H), 6.18 (dd, 7=16.7, 2.3 Hz, 1H), 5.76 (d, 7=10.3, 2.4 Hz, 1H), 3.93 (s, 3H), 3.81-3.93 (m, 4H), 3.58-3.64 (m, 4H). 1-3 450.0 Ή NMR (400 MHz, DMSO-7 ₆ ) δ 9.91 (br. s, 1H), 8.18 (s, 1H), 7.76 (d, 7=8.2 Hz, 1H), 7, 47 (s, 1H), 7.37-7.43 (m, 1H), 7.24-7.29 (m, 1H), 7.17-7.23 (m, 2H), 7.01 ( d, 7=2.4 Hz, 1H), 6.87 (dd, 7=16.7, 10.5 Hz, 1H), 6.19 (dd, 7=16.7, 2.3 Hz, 1H ), 5.73-5.79 (m, 1H), 3.82-3.95 (m, 4H), 3.583.64 (m, 4H). 1-4 464.0 Ή NMR (400 MHz, DMSO-7 ₆ ) δ 8.19 (s, 1H), 7.91 (d, 7=8.4 Hz, 1H), 7.46-7.52 (m, 2H), 7.43 (d, 7=2.4 Hz, 1H), 7.25-7.36 (m, 2H), 7.10 (d, 7=2.5 Hz, 1H), 6.87 (dd , 7=16.7, 10.5 Hz, 1H), 6.31 (dd, 7=1.9, 16.8 Hz, 1H), 5.74-

- 172 044387

5.80 (m, 1H), 3.93 (s, 3H), 3.82-3.93 (m, 4H), 3.56-3.63 (m, 4H). Ή NMR (400 MHz, DMSO-J ₆ ) δ 8.12 (s, 1H), 7.41-7.49 (m, 1H), 7.41 (s, 1H), 7.00 (d, 7 =8.4 Hz, 1H), 6.80-6.92 1-5 432.0 (m, 2H), 6.18 (dd, /=16.8, 2.4 Hz, 1H), 5.75 (dd, 7=10.5, 2.3 Hz, 1H), 3.79- 3.93 (m, 4H), 3.75 (s, 3H), 3.53-3.62 (m, 4H). Ή NMR (400 MHz, DVISOR) δ 9.98 (br s., 1H), 8.10 (s, 1H), 7.39 1-6 418.0 (s, 1H), 7.20-7.29 (m, 1H), 6.68-6.90 (m, 3H), 6.17 (dd, 7=16.6, 2.4 Hz, 1H ), 5.75 (dd, 7=10.4, 2.4 Hz, 1H), 3.783.93 (m, 4H), 3.53-3.58 (m, 4H). Ή NMR (400 MHz, DMSO-d ₆ ) δ 9.79-10.10 (1 H, m), 8.05-8.12 (1 Η, m), 7.77-7.83 (1 Η , m), 7.39-7.48 (1 Η, 1-7 498 m), 7.21-7.29 (3 Η, m), 7.04-7.09 (1 Η, m), 6.76-6.91 (1 Η, m), 6.14-6 .24 (1 Η, m), 5.74-5.81 (1 Η, m), 5.02-5.30 (1 Η, m), 4.08-4.53 (3 Η, m) , 3.54-3.81 (6 Η, m). Ή NMR (400 MHz, DMSO-dY δ 9.83-10.10 (1 Η, m), 8.01-8.07 (1 Η, m), 7.77-7.84 (1 Η, m ), 7.39-7.47 (1 Η, 1-8 496 m), 7.19-7.31 (3 Η, m), 7.04-7.10 (1 Η, m), 6.80-6.94 (1 Η, m), 6.15-6 .28 (1 Η, m), 5.75-5.83 (1 Η, m), 4.38-4.58 (1 Η, m), 4.06-4.27 (2 Η, m) , 3.51-3.89 (3 Η, m), 3.193.29 (1 Η, m), 1.58-1.74 (2 Η, m), 0.90-0.99 (3 Η, m). Ή NMR (400 MHz, DMSO-dY δ 9.85-10.08 (1 H, m), 8.73-8.90 (1 Η, m), 7.77-7.83 (1 Η, m ), 7.72-7.76 (1 Η, 1-9 467.8 m), 7.41-7.47 (1 Η, m), 7.21-7.28 (3 Η, m), 7.04-7.09 (1 Η, m), 6.15-6 .23 (2 Η, m), 5.67-5.72 (1 Η, m), 4.90-5.01 (1 Η, m), 4.36-4.58 (2 Η, m) , 4.16-4.31 (1 Η, m), 1.541.62 (3 Η, m). Ή NMR (400 MHz, DMSO-d ₆ ) δ 13.03-13.21 (1 Η, m), 8.05-8.11 (1 Η, m), 7.53-7.63 (2 Η , m), 7.35-7.41 (1 Η, 1-10 470 m), 6.80-6.99 (1 Η, m), 6.15-6.28 (1 Η, m), 5.74-5.83 (1 Η, m), 4.28-4 .55 (2 Η, m), 4.15-4.28 (1 Η, m), 3.48-3.83 (4 Η, m), 2.16-2.22 (3 Η, m) , 1.17-1.28 (3 Η, m).

- 173 044387

1-11 512 Ή NMR (400 MHz, DMSO-dy δ 9.83-10.06 (1 Η, m), 8.19-8.25 (1 Η, m), 7.76-7.85 (1 Η, m ), 7.38-7.48 (1 Η, m), 7.19-7.30 (3 Η, m), 7.03-7.10 (1 Η, m), 6.81-6, 95 (1 Η, m), 6.15-6.27 (1 Η, m), 5.74-5.81 (1 Η, m), 4.73-4.88 (1 Η, m), 4.36-4.59 (2 Η, m), 4.13-4.26 (1 Η, m), 4.074.14 (1 Η, m), 3.49-3.80 (5 Η, m ), 1.72-1.89 (2 Η, m). 1-12 511 Ή NMR (400 MHz, DMSO-de) δ 8.26-8.34 (1 Η, m), 8.008.09 (2 Η, m), 7.70-7.77 (2 Η, m), 7 .53-7.61 (2 Η, m), 7.41-7.49 (2 Η, m), 7.32-7.41 (2 Η, m), 7.18-7.25 (1 Η, m), 5.15-5.27 (1 Η, m), 4.31-4.56 (2 Η, m), 3.63-3.91 (2 Η, m), 3.42 -3.60 (2 Η, m). 1-13 482.0 Ή NMR (400 MHz, DMSO-X δ 8.13 (d, 7=9.1 Hz, 1H), 8.09 (s, 1H), 7.97 (d, 7=6.8 Hz, 1H) , 7.62 (d, 7=9.0 Hz, 1H), 7.38-7.44 (m, 2H), 7.22 (d, 7=8.8 Hz, 1H), 6.87 ( dd, 7=16.8, 10.6 Hz, 1H), 6.19 (dd, 7=16.8, 2.4 Hz, 1H), 5.77 (dd, 7=10.5, 2, 3 Hz, 1H), 3.82-3.94 (m, 4H), 3.87 (s, 3H), 3.63-3.69 (m, 4H). 1-14 468.0 Ή NMR (400 MHz, DMSO-7 ₆ ) δ 9.91 (br. s, 1H), 8.07 (s, 1H), 7.90 (d, 7=9.0 Hz, 1H), 7, 87 (d, 7=7.8 Hz, 1H), 7.277.38 (m, 3H), 7.15 (d, 7=8.2 Hz, 1H), 6.87 (dd, 7=16.6 ,10.6 Hz, 1H), 6.19 (dd, 7=16.7, 2.3 Hz, 1H), 5.76 (dd, 7=10.5, 2.3 Hz, 1H), 3 .80-3.95 (m, 4H), 3.60-3.67 (m, 4H). 1-15 497.8 Ή NMR (400 MHz, DMSO-dd) δ 8.03-8.12 (1 H, m), 7.68-7.79 (1 Η, m), 7.32-7.44 (1 Η, m), 7.11-7.26 (3 Η, m), 6.98-7.06 (1 Η, m), 6.77-6.91 (1 Η, m), 6.13-6 .26 (1 Η, m), 5.73-5.83 (1 Η, m), 4.08-4.54 (3 Η, m), 3.54-3.80 (7 Η, m) . 1-16 498.0 Ή NMR (400 MHz, DMSO-de) δ 9.88-10.15 (1 Η, m), 8.08-8.16 (1 Η, m), 7.77-7.83 (1 Η, m), 7.40-7.48 (1 Η, m), 7.18-7.30 (3 Η, m), 7.05-7.10 (1 Η, m), 6.80-6 .92 (1 Η, m), 6.15-6.24 (1 Η, m), 5.73-5.80 (1 Η, m), 4.58-4.71 (1 Η, m) , 4.26-4.43 (1 Η, m), 3.99-4.24 (2 Η, m), 3.394.00 (6 Η, m).

- 174 044387

1-17 494.0 Ή NMR (400 MHz, DMSO-de) δ 9.81-10.05 (1 Η, m), 8.06-8.12 (1 Η, m), 7.77-7.84 (1 Η, m), 7.40-7.47 (1 Η, m), 7.19-7.29 (3 Η, m), 7.03-7.08 (1 Η, m), 6.77-6 .88 (1 Η, m), 6.19-6.30 (1 Η, m), 5.73-5.82 (1 Η, m), 4.73-4.86 (2 Η, m) , 3.82-3.96 (2 Η, m), 3.44-3.52 (2 Η, m), 1.882.14 (4 Η, m). 1-18 498.0 Ή NMR (400 MHz, DMSO-de) δ 9.85-10.08 (1 H, m), 8.07-8.13 (1 Η, m), 7.77-7.83 (1 Η, m), 7.40-7.48 (1 Η, m), 7.22-7.30 (3 Η, m), 7.04-7.09 (1 Η, m), 6.76-6 .92 (1 Η, m), 6.15-6.25 (1 Η, m), 5.74-5.81 (1 Η, m), 5.04-5.29 (1 Η, m) , 4.10-4.52 (3 Η, m), 3.53-3.78 (5 Η, m). 1-19 482.0 NMR (400 MHz, DMSO-de) δ ppm 9.90-10.04 (1 H, m), 8.02-8.08 (1 H, m), 7.76-7.82 (1 H, m), 7.38-7.46 (1 Η, m), 7.20-7.30 (3 H, m), 7.04-7.08 (1 H, m), 6.79-6 .95 (1 H, m), 6.14-6.27 (1 H, m), 5.73-5.81 (1 H, m), 4.38-4.55 (3 H, m) , 4.28-4.37 (1 H, m), 4.12-4.26 (1 H, m), 3.463.83 (2 H, m), 1.15-1.21 (3 H, m) 1-19-1 482.0 NMR (400 MHz, DMSO-de) δ ppm 9.83-10.08 (1 Η, m), 8.04-8.11 (1 Η, m), 7.77-7.84 (1 Η, m), 7.38-7.48 (1 Η, m), 7.19-7.31 (3 Η, m), 7.05-7.10 (1 Η, m), 6.81-6 .98 (1 Η, m), 6.16-6.28 (1 Η, m), 5.75-5.83 (1 Η, m), 4.40-4.59 (1 Η, m) , 4.16-4.40 (1 Η, m), 3.96-4.07 (1 Η, m), 3.483.83 (3 Η, m), 3.16-3.29 (1 Η, m), 1.10-1.31 (3 Η, m). 1-19-2 482.0 NMR (400 MHz, DMSO-de) δ ppm 9.83-10.10 (1 H, m), 8.03-8.10 (1 H, m), 7.77-7.83 (1 H, m), 7.38-7.48 (1 Η, m), 7.19-7.31 (3 H, m), 7.05-7.10 (1 H, m), 6.81-6 .98 (1 H, m), 6.15-6.28 (1 H, m), 5.73-5.83 (1 H, m), 4.39-4.56 (2 H, m) , 4.15-4.38 (1 H, m), 3.97-4.10 (1 H, m), 3.483.84 (3 H, m), 1.13-1.27 (3 H, m). 1-20 454.0 ^X H NMR (400 MHz, DMSO-de) δ 9.83-10.05 (1 H, m), 9.11-9.35 (1 Η, m), 8.02-8.07 (1 Η , m), 7.76-7.83 (1 Η, m), 7.40-7.47 (1 Η, m), 7.20-7.28 (3 Η, m), 7.02- 7.09 (1 Η, m), 6.33-6.46 (1 Η, m), 6.11-6.22 (1 Η, m), 5.69-5.78 (1 Η, m ), 4.69-4.80 (1 Η, m), 4.40-4.49 (1 Η, m), 4.314.41 (1 Η, m), 4.21-4.30 (1 Η , m), 3.99-4.09 (1 Η, m).

- 175 044387

1-21 482.0 Ή NMR (400 MHz, DMSO-dX) δ 9.85-10.10 (1 Η, m), 8.04-8.09 (1 Η, m), 7.77-7.83 (1 Η, m), 7.40-7.47 (1 Η, m), 7.19-7.30 (3 Η, m), 7.05-7.09 (1 Η, m), 6.74-6 .99 (1 Η, m), 6.09-6.36 (1 Η, m), 5.66-5.88 (1 Η, m), 4.39-4.57 (2 Η, m) , 4.17-4.39 (1 Η, m), 3.96-4.07 (1 Η, m), 3.483.83 (3 Η, m), 1.10-1.29 (3 Η, m). 1-22 468.0 Ή NMR (400 MHz, DMSO dy δ 9.87-10.03 (1 H, m), 8.45-8.66 (1 Η, m), 7.94-8.01 (1 Η, m) , 7.76-7.83 (1 Η, m), 7.39-7.47 (1 Η, m), 7.21-7.29 (3 Η, m), 7.01-7.10 (1 Η, m), 6.10-6.25 (2 Η, m), 5.59-5.70 (1 Η, m), 4.56-4.68 (1 Η, m), 3 .99-4.13 (1 Η, m), 3.75-3.87 (2 Η, m), 3.553.66 (1 Η, m), 2.35-2.48 (1 Η, m) , 2.06-2.21 (1 Η, m). 1-23 482.0 Ή NMR (400 MHz, DMSO-d ₆ ) δ 9.85-10.06 (1 Η, m), 8.27-8.35 (1 Η, m), 7.95-8.00 (1 Η , m), 7.76-7.84 (1 Η, m), 7.39-7.48 (1 Η, m), 7.20-7.29 (3 Η, m), 7.01- 7.10 (1 Η, m), 6.23-6.37 (1 Η, m), 6.10-6.20 (1 Η, m), 5.58-5.68 (1 Η, m ), 4.00-4.11 (1 Η, m), 3.89-3.99 (1 Η, m), 3.673.77 (1 Η, m), 3.35-3.49 (2 Η , m), 1.94-2.07 (2 Η, m), 1.77-1.90 (1 Η, m), 1.62-1.72 (1 Η, m). 2-7 496.2 NMR (DMSO-d ₆ ) δ: 9.83-10.11 (m, 1H), 7.90-7.95 (m, 1H), 7.76-7.84 (m, 1H), 7, 37-7.49 (m, 1H), 7.19-7.31 (m, 3H), 7.06-7.10 (m, 1H), 6.86-7.00 (m, 1H), 6.22-6.31 (m, 1H), 5.78-5.86 (m, 1H), 4.07-4.22 (m, 3H), 3.89-4.01 (m, 1H ), 3.71-3.84 (m, 1H), 3.40-3.52 (m, 1H), 1.22-1.34 (m, 6H) 2-8 506.0 NMR (DMSO-d ₆ ) δ: 13.04-13.25 (m, 1H), 8.09-8.15 (m, 1H), 7.55-7.62 (m, 2H), 7, 35-7.41 (m, 1H), 6.76-6.94 (m, 1H), 6.36-6.62 (m, 1H), 6.16-6.24 (m, 1H), 5.75-5.83 (m, 1H), 4.59-4.86 (m, 1H), 4.19-4.58 (m, 1H), 3.52-3.92 (m, 3H ), 3.40-3.50 (m, 1H), 3.13-3.27 (m, 1H), 2.13-2.21 (m, 3H) 2-9 486.0 Ή NMR (DMSO-d ₆ ) δ: 10.08-10.30 (m, 1H), 7.98-8.07 (m, 1H), 7.29-7.39 (m, 1H), 6 .72-6.91 (m, 3H), 6.23-6.63 (m, 1H), 6.13-6.24 (m, 1H), 5.74-5.81 (m, 1H) , 4.58-4.82 (m, 1H), 4.42-4.57 (m, 1H), 4.17-4.38 (m, 1H), 3.52-3.89

- 176 044387 (m, 4H)

2-10 510.0 Ή NMR (DMSO-d ₆ ) δ: 9.83-10.06 (m, 1H), 7.93-8.06 (m, 1H), 7.75-7.85 (m, 1H), 7 .39-7.51 (m, 1H), 7.17-7.32 (m, 3H), 7.00-7.15 (m, 1H), 6.76-6.96 (m, 1H) , 6.12-6.30 (m, 1H), 5.68-5.86 (m, 1H), 4.59-4.71 (m, 1H), 4.35-4.47 (m, 1H), 4.06-4.28 (m, 1H), 3.60-4.06 (m, 3H), 3.43-3.58 (m, 1H), 2.08-2.25 ( m, 1H), 1.00-1.11 (m, 3H), 0.86-0.97 (m, 3H) 1-28 436.0 Ή NMR (400 MHz, DMSO-d ₆ ) δ 9.63 (s, 1 Η) 8.04 (s, 1 Η) 7.09-7.24 (m, 1 Η) 6.72-6.95 (m, 3 Η) 6.12-6.24 (m, 1 Η) 5.74-5.77 (m, 1 Η) 3.83-3.88 (d, 4 Η) 3.52-3 .72 (m, 4 Η) 2-1 451.0 Ή NMR (400 MHz, DMSO-J ₆ ) δ 9.97 (br. s, 1H), 8.72 (s, 1H), 7.79 (d, 7=8.6 Hz, 1H), 7, 42 (t, 7=7.1 Hz, 1H), 7.177.28 (m, 3H), 7.09 (d, 7=2.1 Hz, 1H), 6.86 (dd, 7=16.7, 10.5 Hz, 1H), 6.19 (dd, 7=16.7, 2.3 Hz, 1H), 5.74-5.79 (m, 1H), 3.81-3.95 (m , 4H), 3.68-3.76 (m, 4H) 2-2 447.0 Ή NMR (400 MHz, DMSO-7 ₆ ) δ 8.58 (d, 7=3.7 Hz, 1H), 7.47-7.55 (m, 1H), 7.03 (d, 7=8 ,4 Hz, 1H), 6.96 (t, 7=8.7 Hz, 1H), 6.79-6.93 (m, 1H), 6.13-6.24 (m, 1H), 5 .77 (dd, 7=10.5, 2.1 Hz, 1H), 4.26-4.54 (m, 2H), 3.96-4.25 (m, 1H), 3.65-3 .84 (m, 2H), 3.76 (d, 7=2.4 Hz, ZN), 3.47-3.64 (m, 2H), 1.19 (s, 3H). 2-3 433.0 Ή NMR (400 MHz, METHANOL-7/ δ 8.51 (s, 1H), 7.257.34 (m, 1H), 6.74-6.93 (m, 1H), 6.65-6.76 ( m, 2H), 6.31 (d, 7=16.4 Hz, 1H), 5.84 (dd, 7=10.6, 1.5 Hz, 1H), 4.414.53 (m, 2H), 4.03-4.15 (m, 1H), 3.54-3.88 (m, 4H), 1.30 (d, 7=6.6 Hz, ZN). 2-4 479.0 Ή NMR (400 MHz, DMSO-J ₆ ) δ 8.67 (s, 1H), 7.93 (d, 7=8.2 Hz, 1H), 7.45-7.53 (m, 2H), 7.26-7.37 (m, 2H), 7.19 (d, 7=2.5 Hz, 1H), 6.81-6.95 (m, 1H), 6.15-6.22 ( m, 1H), 5.78 (dd, 7=10.4, 2.2 Hz, 1H), 4.39-4.58 (m, 2H), 4.164.26 (m, 1H), 3.94 (s, 3H), 3.68-3.84 (m, 2H), 3.48-3.64 (m, 2H), 1.22 (br. s, 3H).

- 177 044387

2-5 465.0 Ή NMR (400 MHz, DMSO-J ₆ ) δ 9.96 (s, 1H), 8.66 (s, 1H), 7.79 (d, 7=8.4 Hz, 1H), 7.42 ( t, 7=7.0 Hz, 1H), 7.17-7.30 (m, 3H), 7.09 (d, 7=2.4 Hz, 1H), 6.80-6.94 (m , 1H), 6.146.27 (m, 1H), 5.78 (dd, 7=10.6, 2.2 Hz, 1H), 3.97-4.57 (m, 3H), 3.48- 3.83 (m, 4H), 1.22 (br. s, 3H). 2-6 453.0 'NMR (400 MHz, DMSO-7 ₆ )5 13.11 (br. s, 1H), 8.66 (d, 7=2.3 Hz, 1H), 7.55 (d, 7=8.5 Hz, 2H), 7.33 (d, 7=8.5 Hz, 1H), 6.80-6.93 (m, 1H), 6.16-6.24 (m, 1H), 5.78 (dd, 7=10.5, 2.3 Hz, 1H), 3.97-4.56 (m, 4H), 3.48-3.85 (m, 3H), 2.19 (s, 3H ), 1.22 (brs., 3H). 3-1 468.0 Ή NMR (400 MHz, DMSO-de) δ 9.90-10.04 (1H, m), 8.10 (1H, s), 7.80 (1H, d, 7=8.41 Hz), 7 .43 (1H, ddd, 7=1.96, 6.11, 8.17 Hz), 7.16-7.31 (3H, m), 7.07 (1H, d, 7=2.35 Hz ), 6.87 (1H, dd, 7=10.47, 16.73 Hz), 6.19 (1H, dd, 7=2.25, 16.73 Hz), 5.77 (1H, dd, 7=2.25, 10.47 Hz), 3.88 (4H, br d, 7=19.56 Hz), 3.61-3.72 (4H, m). ¹⁹ F NMR (376 MHz, DMSO-d ₆ ) δ -123.78 (s, IF). 3-2 469 Ή NMR (400 MHz, CDC1 ₃ ) δ 9.43 (br. s., 1 Η) 8.12 (d, 7=15.1 Hz, 1 Η) 7.77 (d, 7=9.6 Hz , 1 Η) 7.66 (d, 7=7.6 Hz, 1 Η) 7.59 (d, 7=8.0 Hz, 1 Η) 7.45 (s, 1 Η) 7.21-7 .26 (m, 1 Η) 7.03 (br d, 7=10.4 Hz, 1 Η) 6.95 (d, 7=15.1 Hz, 1 Η) 6.71 (d, 7=9 .6 Hz, 1 Η) 3.93-4.14 (m, 4 Η) 3.52-3.59 (m, 4 Η). ¹⁹ F NMR (376 MHz, CDC1 ₃ ) δ -123.91 (s, 1 F). 3-3 453.0 Ή NMR (400 MHz, DMSO-de) δ 8.82-8.88 (1 Η, m), 8.52-8.63 (1 Η, m), 8.09-8.22 (2 Η, m), 7.89-8.02 (2 Η, m), 7.67-7.74 (1 Η, m), 6.80-6.96 (1 Η, m), 6.13-6 .25 (1 Η, m), 5.72-5.84 (1 Η, m), 3.77-4.03 (4 Η, m), 3.56-3.74 (4 Η, m) . 3-4 469.0 Η NMR (400 MHz, DMSO-de) δ 11.81-12.11 (1 H, m), 8.05-8.15 (1 Η, m), 7.58-7.67 (1 Η, m), 7.42-7.49 (1 Η, m), 7.35-7.42 (1 Η, m), 7.14-7.20 (1 Η, m), 6.80-6 .93 (1 Η, m), 6.44-6.53 (1 Η, m), 6.14-6.24 (1 Η, m), 5.72-5.82 (1 Η, m) , 3.80-3.95 (4 Η, m), 3.61-3.69 (4 Η, m).

- 178 044387

3-5 456.0 Ή NMR (400 MHz, DMSO-d^ δ 12.99-13.26 (1 Η, m), 8.10-8.14 (1 Η, m), 7.54-7.60 (2 Η, m), 7.33-7.40 (1 Η, m), 6.81-6.94 (1 Η, m), 6.14-6.25 (1 Η, m), 5.74-5 .80 (1 Η, m), 3.81-3.95 (4 Η, m), 3.62-3.71 (4 Η, m), 2.12-2.20 (3 Η, m) . 3-6 436.0 Ή NMR (400 MHz, METHANOL-d^ δ 7.84 (d, 7=1.17 Hz, 1H), 7.26 (dt, 7=6.75, 8.27 Hz, 1H), 6.71 -6.90 (m, 2H), 6.62-6.71 (m, 1H), 6.25 (dd, 7=1.96, 16.82 Hz, 1H), 5.79 (dd, 7 =1.86, 10.66 Hz, 1H), 3.91-4.03 (m, 4H), 3.57-3.71 (m, 4H), 3.33 (s, 1H). ¹⁹ F NMR (377 MHz, METHANOL-d^) δ -116.77 (1 F, s), -125.66 (1 F, d, 7=2.6 Hz) 3-7 438.0 Ή NMR (400 MHz, DMSO-A) δ 8.08 (s, 1 Η), 7.53-7.62 (m, 1 Η), 7.47 (td, 7=9.7, 2.5 Hz, 1 Η), 7.28 (td, 7=8.5, 2.2 Hz, 1 Η), 6.85 (dd, 7=16.7, 10.5 Hz, 1 Η), 6, 18 (dd, 7=16.7, 2.2 Hz, 1 Η), 5.71-5.79 (m, 1 Η), 3.86 (br d, 7=19.6 Hz, 4 Η) , 3.63 (t, 7=5.2 Hz, 4 Η) 3-8 432.2 Ή NMR (400 MHz, DMSO-A) δ 9.38 (s, 1 Η), 8.02 (s, 1 Η), 7.15 (d, 7=8.4 Hz, 1 Η), 6, 85 (dd, 7=16.7, 10.5 Hz, 1 Η), 6.77 (dd, 7=8.2, 2.5 Hz, 1 Η), 6.58 (d, 7=2, 5 Hz, 1 Η), 6.18 (dd, 7=16.6, 2.3 Hz, 1 Η), 5.72-5.80 (m, 1 Η), 3.773.93 (m, 4 Η ), 3.62 (t, 7 = 5.1 Hz, 4 Η), 1.94 (s, 3 Η). ¹⁹ F NMR (376 MHz, DMSO-7 ₆ ) δ -124.66 (s, 1 F) 3-9 467.8 Ή NMR (400 MHz, DMSO-de) δ 8.05 (s, 1 Η) 7.54 (d, 7=8.8 Hz, 1 Η) 7.10 (dd, 7=8.8, 3, 1 Hz, 1 Η) 7.02-7.07 (m, 1 Η) 6.85 (dd, 7=16.7, 10.5 Hz, 1 Η) 6.18 (dd, 7=16.7 , 2.2 Hz, 1 Η) 5.72-5.79 (m, 1 Η) 3.86 (br d, 7=19.4 Hz, 4 Η) 3.79 (s, 3 Η) 3, 62 (t, 7=5.1 Hz, 4 N). ¹⁹ F NMR (376 MHz, DMSO-7 ₆ ) δ -123.83 (s, 1 F) 3-10 454.0 Ή NMR (400 MHz, DMSO-d^ δ 10.09 (s, 1 Η), 8.05 (s, 1 Η), 7.39 (dd, 7=11.0, 9.6 Hz, 1 Η ), 6.97 (dd, 7=9.5, 7.1 Hz, 1 Η), 6.85 (dd, 7=16.6, 10.4 Hz, 1 Η), 6.17 (dd, 7=16.6, 2.2 Hz, 1 Η), 5.75 (dd, 7=10.5, 2.2 Hz, 1 Η), 3.76-3.95 (m, 4 Η), 3.62 (br t, 7=5.1 Hz, 4 H) ¹⁹ F NMR (376 MHz, DMSOd ₆ ) δ -122.67 (br s, 1 F), -123.60 (d, 7= 4.3 Hz, 1 F), 130.52 (d, 7=2.6 Hz, IF)

- 179 044387

3-11 453.8 Ή NMR (400 MHz, DMSO-de) δ 9.83-10.02 (1 Η, m), 8.04 (1 Η, s), 7.40 (1 Η, br d, 7=8.6 Hz), 6.76-7.03 (3 H, m), 6.12-6.22 (1 H, m), 5.69-5.82 (1 H, m), 3.86 (4 H, br d, 7=18.6 Hz), 3.63 (4 H, br d, 7=4.7 Hz). ¹⁹ F NMR (376 MHz, DMSO-7 ₆ ) δ -124.09 (s, 1 F) 3-12 431.0 Ή NMR (400 MHz, DMSO dp δ 8.00 (s, 1 Η), 6.99 (d, 7=8.2 Hz, 1 Η), 6.85 (dd, 7=16.6, 10, 4 Hz, 1 Η), 6.57 (dd, 7=8.0, 2.0 Hz, 1 Η), 6.39 (d, 7=1.8 Hz, 1 Η), 6.18 (dd , 7=16.7, 2.2 Hz, 1 Η), 5.69-5.85 (m, 1 Η), 4.99 (s, 2 Η), 3.86 (br d, 7=19 .6 Hz, 4 Η), 3.56-3.70 (m, 4 Η), 1.88 (s, 3 Η). ¹⁹ F NMR (376 MHz, DMSO-de) δ -124.75 (1 F, s) 3-13 473.0 Ή NMR (400 MHz, DMSO dp δ 9.98 (s, 1 Η), 8.05 (s, 1 Η), 7.54 (br d, 7=8.4 Hz, 1 Η), 7.49 (s, 1 Η), 7.29 (d, 7=8.4 Hz, 1 Η), 6.85 (dd, 7=16.5, 10.5 Hz, 1 Η), 6.18 (dd , 7=16.6, 2.0 Hz, 1 Η), 5.76 (dd, 7=10.5, 2.1 Hz, 1 Η), 3.86 (br d, 7=19.8 Hz , 4 Η), 3.63 (br t, 7=4.9 Hz, 4 Η), 2.03 (s, 3 Η), 2.01 (s, 3 H). ¹⁹ F NMR (376 MHz, DMSO~d ₆ ) δ -124.58 (s, IF) 3-14 435.0 Ή NMR (400 MHz, DMSO-de) δ 8.03 (s, 1 Η), 7.01 (t, 7=9.2 Hz, 1 Η), 6.85 (dd, 7=16.7, 10.5 Hz, 1 Η), 6.62-6.71 (m, 1 Η), 6.52 (dd, 7=6.1, 2.7 Hz, 1 Η), 6.18 (dd, 7=16.7, 2.2 Hz, 1 Η), 5.70-5.81 (m, 1 Η), 5.11 (s, 2 Η), 3.86 (br d, 7=19, 6 Hz, 4 Η), 3.52-3.69 (m, 4 H). ¹⁹ F NMR (376 MHz, DMSO dp δ -123.91 (s, 1 F), -131.17 (s, 1 F) 3-15 453.0 Ή NMR (400 MHz, DMSO dp δ 8.06 (s, 1 Η), 6.85 (dd, 7=16.7, 10.5 Hz, 1 Η), 6.63 (ddd, 7=13, 0, 6.6, 2.5 Hz, 1 Η), 6.33 (br d, 7=2.0 Hz, 1 Η), 6.18 (dd, 7=16.6, 2.3 Hz, 1 Η), 5.71-5.82 (m, 1 Η), 5.44 (s, 2 Η), 3.76-3.96 (m, 4 Η), 3.563.71 (m, 4 Η ) 3-16 453.9 Ή NMR (400 MHz, DMSO-d^ δ 10.12 (s, 1 Η), 8.08 (s, 1 Η), 6.92 (ddd, 7=12.3, 6.5, 2.9 Hz, 1 Η), 6.85 (dd, 7=16.6, 10.4 Hz, 1 Η), 6.58-6.65 (m, 1 Η), 6.18 (dd, 7=16 ,7, 2.2 Hz, 1 Η), 5.72-5.79 (m, 1 Η), 3.78-3.92 (m, 4 Η), 3.63 (t, 7=5, 2 Hz, 4 Η)

- 180 044387

3-17 486.0 Ή NMR (400 MHz, DMSO-d^ δ 10.86 (br s, 1 Η), 8.05 (s, 1 Η), 7.68 (s, 1 Η), 6.96 (s, 1 Η ), 6.85 (dd, /=16.6, 10.4 Hz, 1 Η), 6.18 (dd, /=16.6, 2.3 Hz, 1 Η), 5.76 (dd, /=10.5, 2.2 Hz, 1 Η), 3.78-3.93 (m, 4 Η), 3.56-3.68 (m, 4 Η). ¹⁹ F NMR (376 MHz, DMSO-d^δ -123.87 (s, 1 F) 3-18 469.9 Ή NMR (400 MHz, DMSO-d ₆ ) δ 10.36-10.69 (m, 1 Η), 8.04 (s, 1 Η), 7.55 (d, /=11.0 Hz, 1 Η), 6.98 (d, /=9.0 Hz, 1 Η), 6.85 (dd, /=16.7, 10.5 Hz, 1 Η), 6.18 (dd, /=16 ,7, 2.2 Hz, 1 Η), 5.70-5.80 (m, 1 Η), 3.86 (br d, /=19.8 Hz, 4 Η), 3.62 (br t , /=5.1 Hz, 4 H). ¹⁹ F NMR (376 MHz, DMSO-d^ δ -123.82 (s, 1 F), -132.61 (br s, 1 F) 3-19 451.0 ^X H NMR (400 MHz, DMSO-dd) δ 8.01 (s, 1 H), 7.20 (d, /=8.6 Hz, 1 H), 6.85 (dd, /=16.7 , 10.5 Hz, 1 H), 6.66 (dd, /=8.6, 2.7 Hz, 1 H), 6.54 (d, /=2.5 Hz, 1 H), 6, 17 (dd, /=16.7, 2.2 Hz, 1 H), 5.67-5.82 (m, 1 H), 5.40 (s, 2 H), 3.85 (br d, /=19.4 Hz, 4 H), 3.54-3.73 (m, 4 H). ¹⁹ F NMR (376 MHz, DMSO-d^ δ -124.25 (1 F, s) 3-20 485.0 Ή NMR (400 MHz, DMSO-d^ δ 8.04 (s, 1 Η), 7.50 (s, 1 Η), 6.85 (dd, /=16.7, 10.5 Hz, 1 Η ), 6.79 (s, 1 Η), 6.18 (dd, /=16.7, 1.9 Hz, 1 Η), 5.76 (dd, /=10.6, 1.8 Hz, 1 Η), 5.70 (s, 2 Η), 3.86 (br d, /=19.8 Hz, 4 Η), 3.63 (br d, /=4.9 Hz, 4 H). ¹⁹ F NMR (376 MHz, DMSO-J ₆ ) δ -124.05 (br s, 1 F) 3-21 445.2 Ή NMR (400 MHz, DMSO-/ ₆ ) δ 8.54 (d, /=5.4 Hz, 1 Η), 8.29 (s, 1 Η), 8.02 (s, 1 Η), 7 .47 (d, /=5.2 Hz, 1 Η), 6.78 (dd, /=16.7, 10.5 Hz, 1 Η), 6.10 (dd, /=16.7, 2 ,2 Hz, 1 Η), 5.62-5.75 (m, 1 Η), 3.71-3.88 (m, 4 Η), 3.56 (br t, /=5.3 Hz, 4 Η), 2.49-2.64 (m, 1 Η), 1.10 (d, /=6.8 Hz, 3 Η), 1.01 (d, /=6.8 Hz, 3 N ). ¹⁹ F NMR (376 MHz, DMSO-d^ δ 123.04 (1 F, s) 3-22 486.0 Ή NMR (400 MHz, DMSO-/ ₆ ) δ 10.52 (br s, 1 H), 8.07 (s, 1 H), 7.14 (d, /=2.7 Hz, 1 H), 6.78-6.93 (m, 2 H), 6.19 (dd, /=16.7, 2.2 Hz, 1 H), 5.70-5.82 (m, 1 H), 3 .79-3.92 (m, 4 H), 3.63 (br t, /=5.0 Hz, 4 H). ¹⁹ F NMR (376 MHz, DMSO/ ₆ ) δ-124.13 (s, IF)

- 181 044387

3-23 452.1 Ή NMR (400 MHz, DMSO-d ₆ ) δ 8.03-8.14 (m, 3 Η) 7.417.69 (m, 5 Η) 6.84-6.91 (m, 1 Η) 6.17 -6.22 (m, 1 Η) 5.765.79 (m, 1 Η) 3.86-3.91 (m, 4 Η) 3.65-3.67 (m, 4 Η) 3-24 453 Ή NMR (400 MHz, DMSO-d ₆ ) δ 8.78-8.87 (m, 1 Η) 8.438.54 (m, 1 Η) 8.04-8.40 (m, 2 Η) 7.45 -7.79 (m, 2 Η) 7.537.65 (m, 1 Η) 6.74-6.98 (m, 1 Η) 6.09-6.26 (m, 1 Η) 5.795.81 (m , 1 Η) 3.83-3.93 (m, 4 Η) 3.52-3.66 (m, 4 Η) 3-25 442 Ή NMR (400 MHz, DMSO-d ₆ ) δ 8.03 (s, 1 Η) 6.81-6.98 (m, 4 Η) 6.15-6.20 (m, 1 Η) 5.72 -5.86 (m, 3 Η) 3.83-3.88 (m, 4 Η) 3.6-3.62 (m, 4 Η) 4-1 480.0 NMR (400 MHz, DMSO-d^ δ 9.82-10.04 (1 H, m), 7.79 (1 Η, d, 7 = 8.2 Hz), 7.66 (1 H, s), 7.43 (1 H, dt, 7=8.3, 4.0 Hz), 7.26 (1 H, dt, 7=2.3 Hz), 7.22 (2 H, d, 7=3 .7 Hz), 7.05 (1 H, d, 7=2.3 Hz), 6.26-6.38 (1 H, m), 6.12 (1 H, dd, 7=16.8 , 2.2 Hz), 5.66-5.72 (1 H, m), 4.58-4.67 (4 Η, m), 4.50 (2 Η, s), 4.22 (2 Η, s) ^.19 F NMR (376 MHz, DMSO-de) δ-123.98 (1 F, s) 4-2 418.0 Ή NMR (400 MHz, DMSO-d/) δ 7.65 (1 H, d, 7=1.4 Hz), 6.25-6.36 (1 H, m), 6.10 (1 H, dd, 7=17.0, 2.3 Hz), 5.645.72 (1 H, m), 4.58 (4 Η, s), 4.47 (2 Η, s), 4.18 (2 Η , s). ¹⁹ F NMR (376 MHz, DMSO-de) δ -113.54 (1 F, s) 4-3 494.0 Ή NMR (400 MHz, DMSO-de) δ 7.93 (1 H, d, 7=8.4 Hz), 7.67 (1 H, s), 7.45-7.57 (2 H, m ), 7.23-7.36 (2 H, m), 7.16 (1 H, d, 7=2.5 Hz), 6.27-6.39 (1 H, m), 6.11 (1 Η, dd, 7=17.0, 2.2 Hz), 5.65-5.76 (1 N, m), 4.58-4.67 (4 H, m), 4.50 ( 2H, s), 4.22 (2H, s), 3.93 (3H, s). ¹⁹ F NMR (376 MHz, DMSO-de) δ -123.88 (1 F, s) 4-4 391.8 Ή NMR (400 MHz, DMSO-de) δ 8.93 (1 Η, br s), 7.637.77 (1 Η, m), 6.04-6.33 (2 Η, m), 5.60- 5.77 (1 Η, m), 4.89 (1 Η, br d, 7=3.3 Hz), 4.72 (2 H, br dd, 7=8.1, 3.6 Hz), 4.28 (2 H, br dd, 7=8.0, 3.9 Hz). ¹⁹ F NMR (377 MHz, DMSOd ₆ ) δ -113.24(1 F, s) 4-5 468.0 1H NMR (400 MHz, DMSO-d6) δ 8.92-9.00 (1 H, m), 7.93 (1 H, d, 7=8.2 Hz), 7.72 (1 H, s ), 7.45-7.60 (2 Η, m), 7.25-

- 182 044387

7.36 (2 H, m), 7.17 (1 Η, d, 7 = 2.3 Hz), 6.08-6.36 (2 H, m), 5.69 (1 H, dd, /=9.8, 2.2 Hz), 4.87-5.01 (1 H, m), 4.694.84 (2 Η, m), 4.33 (2 Η, br d, /=3, 3 Hz), 3.94 (3 H, s). ¹⁹ F NMR (376 MHz, DMSO-de) δ -123.93 (1 F, s) Ή NMR (400 MHz, DMSO-de) δ 9.95 (1 H, s), 8.97 (1 Η, d, /=7.0 Hz), 7.80 (1 H, d, /=8.2 Hz), 7.72 (1 H, s), 7.39- 4-6 454.0 7.49 (1 Η, m), 7.16-7.32 (3 Η, m), 7.05 (1 Η, d, /=2.2 Hz), 6.08-6.36 (2 H, m), 5.65-5.73 (1 H, m), 4.87-5.05 (1 H, m), 4.77 (2 H, td, /=8.2, 2, 6 Hz), 4.33 (2 H, br t, /=6.2 Hz). ¹⁹ F NMR (376 MHz, DMSO-de) δ -124.03 (1 F, s) Ή NMR (400 MHz, DMSO-de) δ 9.23-9.42 (1 H, m), 8.04 (1 Η, d, /=1.0 Hz), 6.37 (1 N, dd, /=17.0, 10.2 Hz), 6.15 (1 4-7 391.8 H, dd, /=17.0, 2.0 Hz), 5.65-5.86 (1 H, m), 4.70 (1 H, br t, /=8.0 Hz), 4, 35-4.48 (1 H, m), 4.25-4.33 (1 H, m), 4.22 (1 H, br dd, /=9.0, 4.3 Hz), 3, 99 (1 H, br dd, /=10.3, 4.4 Hz). ¹⁹ F NMR (376 MHz, DMSO-de) δ -113.81 (1 F, s) Ή NMR (400 MHz, DMSO-de) δ 8.64 (1 H, br s), 8.15 (1 H , br d, /=8.8 Hz), 6.58-6.97 (1 H, m), 5.97-6.27 (1 H, m), 5.58-5.74 (1 H , m), 4.50 (1 H, br d, /=11.5 Hz), 3.91 (1 H, 4-8 420.0 br d, /=13.5 Hz), 3.14-3.28 (2 H, m), 2.85-2.97 (1 H, m), 2.08-2.23 (1 H, m), 1.79-1.91 (1 H, m), 1.64-1.78 (1 H, m), 1.45-1.60 (1 H, m). ¹⁹ F NMR (376 MHz, DMSO-de) δ -114.17(1 F, br s) ^X H NMR (400 MHz, DMSO-de) δ 9.87-10.07 (1 H, m), 8 .50-8.70 (1 H, m), 8.06-8.23 (1 H, m), 7.70-7.88 (1 H, m), 7.39-7.48 (1 H, m), 7.19-7.28 (3 H, m), 7.02-7.08 (1 4-9 482.0 H, m), 6.68-6.93 (1 H, m), 6.05-6.23 (1 H, m), 5.64-5.78 (1 H, m), 4.51 -4.63 (1 H, m), 4.06-4.17 (1 H, m), 3.914.04 (1 H, m), 3.14-3.30 (1 H, m), 2 .91-3.01 (1 H, m), 2.14-2.29 (1 H, m), 1.83-1.96 (1 H, m), 1.68-1.82 (1 H, m), 1.49-1.65 (1 H, m). NMR (400 MHz, DMSO-de) δ 9.89-10.10 (m, 1 H), 7.79 5-1 468.0 (d, /=8.4 Hz, 1 H), 7.73 (s, 1 H), 7.43 (ddd, /=8.2, 5.1, 2.9 Hz, 1 H), 7 ,20-7.30 (m, 3 H), 7.05 (d, /=2.2 Hz, 1 H), 6.81 (dd, /=16.7, 10.5 Hz, 1 H) , 6.10-6.23 (m, 1 H), 5.69-5.81

- 183 044387

(m, 1 Η), 5.37-5.59 (m, 1 Η), 4.63-4.74 (m, 3 Η), 4.534.61 (m, 1 Η), 3.14-3 .23 (m, 3 H). ¹⁹ F NMR (376 MHz, DMSO-de) δ -124.10 (1 F, s) 5-2 468.2 NMR (400 MHz, DMSO-d ₆ ) δ 9.89-10.09 (m, 1 Η), 8.70 (s, 1 Η), 7.79 (d, /=8.2 Hz, 1 Η) , 7.69 (s, 1 Η), 7.39-7.46 (m, 1 Η), 7.16-7.31 (m, 3 Η), 7.05 (d, /=2.2 Hz, 1 Η), 6.206.32 (m, 1 Η), 6.08-6.18 (m, 1 Η), 5.65 (dd, /=10.1, 1.9 Hz, 1 Η) , 4.57 (dd, /=8.1, 1.9 Hz, 2 Η), 4.40 (br d, /=8.4 Hz, 2 Η), 1.67 (s, 3 Η). ¹⁹ F NMR (376 MHz, DMSO-d ₆ ) δ 124.13 (1 F, s) 5-3 484.0 Ή NMR (400 MHz, DMSO-dY δ 9.95 (s, 1 Η), 8.74 (s, 1 Η), 7.79 (d, /=8.4 Hz, 1 Η), 7.70 (s, 1 Η), 7.42 (br t, /=6.6 Hz, 1 Η), 7.16-7.28 (m, 3 Η), 7.05 (d, /=2.2 Hz, 1 Η), 6.236.41 (m, 1 Η), 6.07-6.19 (m, 1 Η), 5.66 (dd, /=10.1, 1.7 Hz, 1 Η) , 5.36 (br t, /=5.8 Hz, 1 Η), 4.49 (s, 4 Η), 3.74 (br d, /=5.5 Hz, 2 H). ¹⁹ F NMR (376 MHz, DMSO dy δ 124.12(1 F, s) 5-4 482.1 Ή NMR (400 MHz, DM SO dp δ 9.99 (s, 1 Η), 8.02 (s, 1 Η), 7.80 (d, /=8.1 Hz, 1 Η), 7.38 -7.52 (m, 1 Η), 7.16-7.34 (m, 3 Η), 7.07 (s, 1 Η), 6.75-6.96 (m, 1 Η), 6 ,11-6.35 (m, 1 Η), 5.66-5.90 (m, 1 Η), 4.43-4.91 (m, 1 Η), 4.07-4.39 (m , 1 Η), 3.90-4.05 (m, 1 Η), 3.71-3.87 (m, 1 Η), 3.61 (br d, /=9.7 Hz, 1 Η) , 3.33-3.51 (m, 2 Η), 1.36 (br s, 3 H) ^.19 F NMR (376 MHz, DMSO dy δ -124.10 (1 F, s) 5-5 480.1 Ή NMR (400 MHz, DMSO-/ ₆ ) δ 9.97 (s, 1 Η), 7.80 (d, /=8.3 Hz, 1 Η), 7.69 (d, /=4.4 Hz, 1 Η), 7.38-7.49 (m, 1 Η), 7.16-7.29 (m, 3 Η), 7.05 (d, /=1.0 Hz, 1 Η) , 6.54-6.86 (m, 1 Η), 6.14-6.31 (m, 1 Η), 5.74-5.80 (m, 1 Η), 5.215.40 (m, 1 Η), 4.96-5.17 (m, 1 Η), 4.61-4.73 (m, 1 Η), 4.15-4.42 (m, 2 Η), 3.55-3 .92 (m, 1 Η), 2.25-2.47 (m, 1 Η), 1.93-2.20 (m, 1 Η) 5-6 480.1 Ή NMR (400 MHz, DMSO-/ ₆ ) δ 9.97 (s, 1 Η), 7.80 (d, /=8.1 Hz, 1 Η), 7.69 (d, /=4.1 Hz, 1 Η), 7.43 (dt, /=8.1, 4.1 Hz, 1 Η), 7.26 (d, /=2.3 Hz, 1 Η), 7.19-7, 25 (m, 2 Η), 7.05 (d, /=2.1 Hz, 1 Η), 6.55-6.83 (m, 1 Η), 6.23 (ddd,

- 184 044387

7=16.5, 7.5, 2.2 Hz, 1 Η), 5.79 (br d, 7=2.1 Hz, 1 Η), 5.225.40 (m, 1 Η), 4.96 -5.18 (m, 1 Η), 4.61-4.72 (m, 1 Η), 4.18-4.44 (m, 2 Η), 3.57-3.94 (m, 1 Η), 2.27-2.45 (m, 1 Η), 1.92-2.24 (m, 1 Η) 5-7 482.1 Ή NMR (400 MHz, DMSO-d^ δ 9.97 (1 H, br s), 8.02 (1 H, s), 7.80 (1 H, d, 7=8.5 Hz), 7 .35-7.53 (1 H, m), 7.177.33 (3 Η, m), 7.07 (1 Η, d, 7 = 2.3 Hz), 6.84 (1 H, dd, 7 =16.7, 10.5 Hz), 6.19 (1 H, dd, 7 = 16.8, 1.7 Hz), 5.68-5.83 (1 H, m), 4.43- 4.93 (1 H, m), 4.08-4.39 (1 H, m), 3.934.05 (1 H, m), 3.79 (1 H, br d, 7 = 11.6 Hz ), 3.54-3.66 (1 Η, m), 3.37-3.50 (2 H, m), 1.35 (3 H, br s). ¹⁹ F NMR (376 MHz, DMSO- de) δ -123.81 (1 F, d, 7=16.5 Hz) 5-8 480.0 Ή NMR (400 MHz, DMSO-7 ₆ ) δ 9.98 (s, 1 Η), 7.86 (br d, 7 = 8.7 Hz, 1 Η), 7.80 (d, 7 = 8, 1 Hz, 1 Η), 7.37-7.53 (m, 1 Η), 7.19-7.31 (m, 3 Η), 7.06 (br s, 1 H), 6.25- 6.55 (m, 1 H), 6.11-6.24 (m, 1 H), 5.64-5.82 (m, 1 H), 5.14-5.44 (m, 1 H ), 5.03-5.12 (m, 1 H), 4.24-4.59 (m, 1 H), 3.63-4.20 (m, 3 H), 2.22-2, 48 (m, 2 H). ¹⁹ F NMR (376 MHz, DMSOJ ₆ ) δ-124.17--123.93 (m, IF) 5-9 468.0 Ή NMR (400 MHz, DMSO-7 ₆ ) δ 9.98 (s, 1 Η), 8.04 (s, 1 Η), 7.80 (d, 7=8.5 Hz, 1 Η), 7 .38-7.52 (m, 1 Η), 7.15-7.35 (m, 3 Η), 7.07 (d, 7=2.3 Hz, 1 Η), 6.39 (dd, 7=17.0, 10.4 Hz, 1 Η), 6.16 (dd, 7=17.0, 2.1 Hz, 1 Η), 5.66-5.81 (m, 1 Η), 4.94-5.06 (m, 1 Η), 4.67 (br t, 7=8.7 Hz, 1 Η), 4.49 (br dd, 7=9.4, 5.3 Hz, 1 Η), 4.31-4.43 (m, 1 Η), 4.21 (br dd, 7=10.6, 5.2 Hz, 1 Η), 3.43 (s, 3 H). ¹⁹ F NMR (376 MHz, DMSO-Tb) δ -124.00 (s, 1 F) 6-1 452.0 Ή NMR (400 MHz, DMSO-d _e ) δ 7.97-8.10 (m, 1 Η), 7.60 (d, 7=8.9 Hz, 1 Η), 6.86 (dd, 7 =16.6, 10.6 Hz, 1 Η), 6.57 (d, 7=8.9 Hz, 1 Η), 6.38 (s, 2 Η), 6.19 (dd, 7=16 ,8, 2.3 Hz, 1 Η), 5.71-5.84 (m, 1 Η), 3.86 (br d, 7=19.9 Hz, 4 Η), 3.63 (br d , 7=1.0 Hz, 4 H). ¹⁹ F NMR (376 MHz, DMSO-d ₆ ) δ 126.04 (1 F, s)

- 185 044387

6-2 437.0 Ή NMR (400 MHz, DMSO-df) δ 8.67-8.79 (1 Η, m), 8.18 (1 Η, dd, 7=8.3, 1.0 Hz), 8.10 ( 1 Η, s), 7.61 (1 Η, dd, 7=8.2, 4.7 Hz), 6.86 (1 N, dd, 7=16.6, 10.4 Hz), 6, 19 (1 H, dd, 7=16.8, 2.3 Hz), 5.77 (1 H, dd, 7=10.4, 2.3 Hz), 3.79-3.97 (4 H , m), 3.58-3.73 (4 Η, m). ¹⁹ F NMR (376 MHz, DMSOd ₆ ) δ -125.75 (1 F, s) 7-1 518.0 Ή NMR (400 MHz, DMSO-7 ₆ ) δ 10.13 (br. s., 1 H) 8.12 (d, 7 = 2.2 Hz, 1 H) 7.80 (d, 7 = 8, 2 Hz, 1 H) 7.43 (br t, 7=7.0 Hz, 1 H) 7.20-7.30 (m, 3 H) 7.08 (dd, 7=5.8, 2, 2 Hz, 1 H) 6.78-6.91 (m, 1 H) 6.27-6.70 (m, 1 H) 6.20 (dd, 7=16.6, 2.0 Hz, 1 H) 5.76-5.84 (m, 1 H) 4.73-4.87 (m, 1 H) 4.194.72 (m, 2 H) 3.55-3.90 (m, 3 H) 3.36-3.47 (m, 1H) 7-2 500.0 Ή NMR (DMSO-d ₆ ) δ: 9.73-10.17 (m, 1H), 8.04-8.12 (m, 1H), 7.77-7.84 (m, 1H), 7 .39-7.48 (m, 1H), 7.20-7.30 (m, 3H), 7.06-7.10 (m, 1H), 6.77-6.93 (m, 1H) , 6.15-6.24 (m, 1H), 5.74-5.83 (m, 1H), 4.57-4.92 (m, 3H), 4.14-4.54 (m, 2H), 3.55-3.87 (m, 3H), 3.21-3.29 (m, 1H) 7-3 526.0 Ή NMR (DMSO-d ₆ ) δ: 9.76-10.22 (m, 1H), 8.09-8.14 (m, 1H), 7.77-7.84 (m, 1H), 7 .39-7.48 (m, 1H), 7.20-7.29 (m, 3H), 7.04-7.11 (m, 1H), 6.82 (br. s., 1H), 6.14-6.22 (m, 1H), 5.77-5.83 (m, 1H), 3.68-5.36 (m, 4H), 3.60-3.67 (m, 3H ) 8-1 547 NMR (400 MHz, DMSCM,) δ 10.06 (br. d., 7 = 15.1 Hz, 1 H) 8.03 (d, 7 = 1.2 Hz, 1 H) 7.51-7, 56 (m, 1 H) 7.45 (t, 7=7.6 Hz, 1 H) 7.33 (tdd, 7=7.5, 7.5, 3.8, 1.4 Hz, 1 H ) 7.147.25 (m, 2 H) 6.84 (dd, 7=16.8, 10.4 Hz, 1 H) 6.62-6.74 (m, 2 H) 6.14-6.26 (m, 2 H) 5.71-5.78 (m, 1 H) 3.71-3.99 (m, 8 H) 2.52-2.59 (m, 1 H) 1.02-1 .12 (m, 6 H). ¹⁹ F NMR (377 MHz, DMSO-7 ₆ ) δ -113.6 (s, 1 F) -114.8 (s, 1 F). 8-2 548.2 ^X NMR (400 MHz, CDC1 ₃ ) δ 8.61 (1 H, br s) 8.17 (1 H, s) 7.49-7.55 (2 H, m) 7.35-7.43 ( 1 H, m) 7.23-7.30 (1 H, m) 7.09 (1 H, d, 7=7.88 Hz) 6.58-6.72 (3 H, m) 6.41 (1 H, dd, 7=16.79, 1.66 Hz) 5.82 (1 H, dd, 7=10.47, 1.55 Hz) 3.804.15 (8 H, m) 2.71 ( 1 H, spt, 7=6.84 Hz) 1.23 (3 H, d, 7=6.84 Hz) 1.03 (4 H, d, 7=6.84 Hz)

- 186044387

8-3 561 Ή NMR (400 MHz, CDC1 ₃ ) δ 7.81 (s, 1 Η) 7.35-7.49 (m, 2 Η) 7.18-7.32 (m, 1 Η) 7.01-7 .17 (m, 2 Η) 6.67-6.74 (m, 1 Η) 6.48-6.65 (m, 3 Η) 6.32-6.44 (m, 1 Η) 5.77 -5.83 (m, 1 Η) 4.19-5.14 (m, 3 Η) 3.75-3.98 (m, 1 Η) 3.41-3.68 (m, 2 Η) 2 .85-3.28 (m, 1 Η) 2.49-2.71 (m, 1 Η) 1.34-1.54 (m, 3 Η) 1.13-1.21 (m, 3 Η ) 1.00-1.07 (m, 3 Η). ¹⁹ NMR (377 MHz, CDC1 ₃ ) δ -113.43 - -113.3 (m, 1 F) -114.3 - -113.9 (m, 1 F). 8-3-1 561.2 Ή NMR (400 MHz, CDC1 ₃ ) δ 7.82 (s, 1 Η) 7.37-7.51 (m, 2 Η) 7.21-7.34 (m, 1 Η) 7.05-7 .20 (m, 2 Η) 6.70 (br d, 7=8.1 Hz, 1 H) 6.52-6.67 (m, 3 H) 6.39 (dd, 7=16.8, 1.7 Hz, 1 H) 5.80 (dd, 7=10.5, 1.6 Hz, 1 H) 4.26-5.00 (m, 3 H) 3.40-4.00 (m , 3 H) 3.06-3.24 (m, 1 H) 2.52-2.69 (m, 1 H) 1.20 (d, 7=6.2 Hz, 6 H) 1.04 ( br d, 7=6.4 Hz, 3 H). ¹⁹ F NMR (377 MHz, CDC1 ₃ ) δ -113.31 (br d, 7=63.3 Hz, 1 F) 113.99 (br d, 7=33.8 Hz, 1 F). 8-3-2 561.2 Ή NMR (400 MHz, CDC1 ₃ ) δ 7.79 (br s, 1 H) 7.35-7.48 (m, 2 H) 7.21-7.32 (m, 1 H) 7.04- 7.17 (m, 2 H) 6.68 (d, 7=8.3 Hz, 1 H) 6.49-6.66 (m, 3 H) 6.36 (br d, 7=16.6 Hz, 1 H) 5.78 (dd, 7=10.4, 1.9 Hz, 1 H) 4.29-5.11 (m, 2 H) 3.46-4.02 (m, 3 H ) 2.93-3.29 (m, 2 H) 2.49-2.68 (m, 1 H) 1.48 (dd, 7=14.5, 2.1 Hz, 6 H) 1.03 (br d, 7=6.0 Hz, 3 H). ¹⁹ F NMR (377 MHz, CDC1 ₃ ) δ -113.32 (br d, 7=9.5 Hz, 1 F) -114.02--113.78 (m, IF). 8-4 576 Ή NMR (400 MHz, CDC1 ₃ ) δ 8.58 (s, 1 Η) 8.13 (s, 1 Η) 7.41-7.50 (m, 1 Η) 7.32 (s, 1 Η) 7.30 (s, 1 Η) 7.22-7.30 (m, 1 Η) 6.53-6.75 (m, 3 Η) 6.42 (dd, 7=16.8, 1.7 Hz, 1 Η) 5.77-5.86 (m, 1 Η) 3.53-5.25 (m, 6 Η) 2.98-3.34 (m, 1 Η) 2.16-2, 49 (m, 4 Η) 1.52 (br d, 7 = 19.5 Hz, 3 Η) 1.07-1.17 (m, 6 Η). ¹⁹ F NMR (377 MHz, CDC1 ₃ ) δ -104.8 (br. s., 1 F) -104.9 (br. s., 1 F). 8-5 546.2 Ή NMR (400 MHz, DMSO-d ₆ ) δ 10.08-10.10 (m, 1 Η) 8.47-8.48 (m, 1 Η) 8.37 (s, 1H), 8.05 (s, 1H), 7.16-7.26 (m, 1H) 7.04-7.25 (m, 1 H) 6.82-6.90 (m, 1 H) 6.63-6, 77 (m, 2 H) 6.30-6.31 (m, 1 H) 6.16-6.21

- 187 044387

(m, 1 Η) 5.72-5.79 (m, 1 Η) 3.84-4.02 (m, 6 Η) 3.76-3.81 (m, 2 Η), 1.54- 1.55 (m, 1 Η) 1.23 (s, 2H) 0.74-0.85 (m, 1 Η), 0.52-0.69 (m, 1 Η) ^X H NMR (400 MHz , CDC1 ₃ ) δ 8.62 (1 H, br s) 8.10-8.13 (1 H, m) 7.49-7.56 (2 H, m) 7.35-7.43 (1 H, m) 7.22-7.30 (1 8-6 562.1 H, m) 7.10 (1 H, br s) 6.54-6.73 (3 H, m) 6.38-6.45 (1 H, m) 5.82 (1 H, dd, 7 =10.57, 1.45 Hz) 2.60-5.27 (8 H, m) 1.42-1.54 (3 H, m) 1.23 (3 H, d, 7 = 6.84 Hz) 1.03 (3 H, d, 7=6.84 Hz) ^X H NMR (400 MHz, DMSO-7 ₆ ) δ 10.42 (br d, 7=17.0 Hz, 1 H), 7 .86-8.11 (m, 1 H), 7.50-7.63 (m, 1 H), 7.47 (br t, 7=6.0 Hz, 1 H), 7.36 (t , 7=7.5 Hz, 1 H), 7.15-7.26 (m, 1 H), 9-1 611.0 7.05 (d, 7=2.3 Hz, 1 H), 6.78-6.96 (m, 1 H), 6.44-6.58 (m, 1 H), 6.11-6 .29 (m, 2 H), 5.71-5.82 (m, 1 H), 4.68-4.98 (m, 1 H), 3.96-4.52 (m, 3 H) , 3.52-3.85 (m, 2 H), 3.343.51 (m, 1 H), 2.95-3.26 (m, 1 H), 1.27-1.41 (m, 3 H), 0.95-1.13 (m, 6H) ^X H NMR (400 MHz, DMSO-7 ₆ ) δ 7.91-8.08 (m, 1 H), 7.49-7.67 ( m, 2 H), 7.41 (br d, 7=5.8 Hz, 1 H), 7.21 (br s, 1 H), 6.76-6.98 (m, 1 H), 6 .52-6.67 (m, 1 H), 6.09-6.29 9-2 531.0 (m, 1 H), 5.75 (br s, 1 H), 4.61-4.96 (m, 1 H), 4.23-4.48 (m, 1 H), 3.93- 4.21 (m, 2H), 3.50-3.77 (m, 1H), 3.33-3.49 (m, 1 H), 3.23-3.28 (m, 1 H), 2.94-3.24 (m, 1 H), 1.27 (br d, 7=9.3 Hz, 6 H), 1.09 (br s, 3 H) ^X H NMR (400 MHz, DMSO -7 ₆ ) δ 7.88-8.01 (m, 1 H), 7.49-7.61 (m, 1 H), 7.46 (br t, 7 = 7.6 Hz, 1 H) , 7.36 (t, 7=7.3 Hz, 1 H), 7.15-7.25 (m, 1 H), 7.08 (d, 7=8.7 Hz, 1 9-3 576.2 H), 6.76-6.95 (m, 1 H), 6.57 (dd, 7=8.7, 2.5 Hz, 1 H), 6.176.32 (m, 2H), 6.11 -6.16 (m, 1 H), 5.72-5.81 (m, 1 H), 5.40 (br d, 7=10.8 Hz, 2 H), 4.66-4.99 (m, 1 H), 4.21-4.52 (m, 2 H), 3.94-4.20 (m, 2 H), 3.53-3.82 (m, 2 H), 3 .36-3.51 (m, 1 H), 1.27-1.40 (m, 3 H), 0.95-1.12 (m, 6 H) ^X H NMR (400 MHz, DMSO-d ₆ ) δ 9.94 (br. s, 1H), 8.10 (s, 9-4 593.2 1H), 7.72 (d, 7=8.1 Hz, 1H), 7.43-7.51 (m, 1H), 7.31-7.42 (m, 3H), 7.11-7 .27 (m, 4H), 6.68-6.93 (m, 2H), 6.14-6.36

- 188 044387

(m, 2H), 5.70-5.83 (m, 1H), 3.72-4.12 (m, 8H), 2.23-2.40 (m, 1H), 1.32-1 .65 (m, 2H), 0.95-1.16 (m, 3H), 0.36-0.75 (m, 3H). 9-5 530.2 Ή NMR (400 MHz, DMSO-d ₆ ) δ 7.97 (s, 1 Η) 7.85-7.90 (m, 2 Η) 7.64-7.72 (m, 2 Η) 7.107.18 (m, 1 Η) 6.73-6.78 (m, 1 Η) 6.57-6.67 (m, 2 Η) 6.33 (s, 1 Η) 6.10-6.14 (m , 1 Η) 5.66-5.72 (m, 1 Η) 3.85-3.96 (m, 4 Η) 3.48-3.79 (m, 2 Η) 3.68-3.75 (m, 2 Η) 9-6 546.2 Ή NMR (400 MHz, DMSO-d ₆ ) δ 9.92-10.25 (m, 1 Η) 8.44-8.82 (m, 1 Η) 8.27-8.40 (m, 1 Η ) 7.99 (s, 1 Η) 7.27-7.28 (m, 1 Η) 7.11-7.21 (m, 1 Η) 6.726.91 (m, 1 Η) 6.48-6 .71 (m, 2 Η) 6.03-6.31 (m, 2 Η) 5.485.79 (m, 1 Η) 3.57 4.16 (m, 8 Η) 1.36-1.70 ( m, 1 Η) 0.210.92 (m, 4 Η) 9-7 561.2 Ή NMR (400 MHz, METAHOJI-d ₄ ) δ 8.44-8.56 (m, 1 Η) 8.41-8.44 (m, 1 Η) 8.00-8.03 (m, 1 Η ) 7.35-7.37 (m, 1 Η) 7.14-7.23 (m, 1 Η) 6.75-6.83 (m, 1 Η), 6.56-6.66 (m , 2 Η) 6.40-6.46 (m, 1 Η) 6.25-6.31 (m, 1 Η) 5.75-5.82 (m, 1 Η) 4.42-4.67 (m, 2 Η) 3.59-3.89 (m, 2 Η) 1.57-1.70 (m, 1 Η) 1.45-1.52 (m, 3 Η) 1.27 (s , 4H) 0.80-0.88 (m, 3 Η) 9-8 561.2 Ή NMR (400 MHz, METAHOJI-d ₄ ) δ 8.43-8.50 (m, 1 Η) 8.30-8.36 (m, 1 Η) 7.91-7.97 (m, 1 Η ) 7.25-7.32 (m, 1 Η) 7.07-7.14 (m, 1 Η) 6.70-6.78 (m, 1 Η) 6.47-6.60 (m, 2 Η) 6.33-6.41 (m, 1 Η) 6.16-6.26 (m, 1 Η) 5.68-5.77 (m, 1 Η) 4.32-4.52 ( m, 2 Η) 3.50-3.81 (m, 2 Η) 1.51-1.62 (m, 1 Η) 1.36 -1.41 (m, 3 Η) 1.19 (s, 4 Η) 0.74-0.85 (m, 3 Η) 9-9 581.2 Ή NMR (400 MHz, METAHOJI-d ₄ ) δ 8.48-8.57 (m, 1 Η) 8.31-8.41 (m, 1 Η) 8.13-8.18 (m, 1 Η ) 7.38-7.54 (m, 3 Η) 7.28-7.40 (m, 1 Η) 6.82-6.95 (m, 1 Η) 6.45 -6.50 (m, 1 Η) 6.28-6.38 (m, 1 Η) 5.81-5.90 (m, 1 Η) 4.47-4.63 (m, 2 Η) 4.07-4.25 ( m, 1 Η) 3.63-3.94 (m, 2 Η) 2.13-2.21 (m, 3 Η) 1.62-1.75 (m, 1 Η) 1.51-1, 57 (m, 3 Η) 1.27-1.37 (m, 3 Η) 0.86-0.95 (m, 3 Η)

- 189 044387

9-10 597.2 Ή NMR (400 MHz, DMSO-J ₆ ) δ 8.01 (br d, 7=18.0 Hz, 1 Η), 7.71 (d, 7=8.1 Hz, 1 Η), 7.50 -7.58 (m, 1 Η), 7.39-7.49 (m, 2 Η), 7.35 (t, 7 = 7.4 Hz, 1 Η), 7.09-7.27 ( m, 2 Η), 6.786.97 (m, 1 Η), 6.14-6.29 (m, 2 Η), 5.71-5.82 (m, 1 Η), 4.70-4, 98 (m, 1 Η), 3.98-4.54 (m, 3 Η), 3.39-3.85 (m, 2 Η), 2.91-3.29 (m, 1 Η), 1.27-1.41 (m, 3 Η), 1.10 (br t, 7=6.2 Hz, 3 Η), 0.94-1.07 (m, 3 Η) 9-11 561.2 Ή NMR (400 MHz, DMSO-7 ₆ ) δ 7.99 (br d, 7 = 16.6 Hz, 1 Η), 7.49-7.68 (m, 2 Η), 7.29-7, 48 (m, 4 Η), 7.10-7.26 (m, 2 Η), 6.76-6.96 (m, 1 Η), 6.12-6.31 (m, 2 Η), 5.71-5.82 (m, 1 Η), 4.68-4.97 (m, 1 Η), 3.95-4.51 (m, 3 Η), 3.443.85 (m, 2 Η ), 2.92-3.26 (m, 2 Η), 1.27-1.41 (m, 3 Η), 1.06-1.17 (m, 3 Η), 0.92-1, 06 (m, 3 Η) 9-12 583 Ή NMR (400 MHz, CDC1 ₃ ) δ 8.35 (s, 1 Η) 8.08 (d, 7 = 8.3 Hz, 1 Η) 7.51-7.59 (m, 1 Η) 7, 34-7.44 (m, 3 Η) 7.21 (td, 7=7.7, 0.8 Hz, 1 Η) 6.85 (d, 7=8.5 Hz, 1 Η) 6.56 -6.75 (m, 1 Η) 6.45 (dd, 7=16.8, 1.7 Hz, 1 Η) 5.81-5.90 (m, 1 Η) 4.33-5.25 (m, 3 Η) 3.82-4.01 (m, 1 Η) 3.05-3.71 (m, 3 Η) 2.40-2.58 (m, 2 Η) 2.20-2 .37 (m, 2 Η) 1.57 (br d, 7=18.0 Hz, 3 Η) 1.10 (td, 7=7.6, 0.8 Hz, 6 Η). 9-13 579.2 Ή NMR (400 MHz, CDC1 ₃ ) δ 7.93 (0.5 Η, s) 7.92 (0.5 Η, s) 7.69-7.74 (1 Η, m) 7.18-7 .51 (7 Η, m) 7.11-7.16 (1 Η, m) 6.88 (1 Η, dd, 7=25.82, 2.54 Hz) 6.59-6.68 (2 Η, m) 6.42 (1 Η, dd, 7=16.82, 1.76 Hz) 5.82 (1 N, dd, 7=10.56, 1.56 Hz) 3.81-4, 11 (8 H, m) 2.65-2.74 (1 Η, m) 1.25 (1.5 Η, d, 7=6.85 Hz) 1.22 (1.5 N, d, 7 =6.85 Hz) 1.13 (1.5 H, d, 7 = 6.85 Hz) 0.98 (1.5 H, d, 7 = 6.85 Hz) 9-14 540.2 Ή NMR (400 MHz, DMSO-Tg) δ 10.21 (0.6 H, br s) 10.12 (0.4 H, br s) 8.29-8.35 (1 H, m) 7, 28-7.38 (1 H, m) 6.736.85 (3 H, m) 6.17 (1 H, dd, 7=16.59, 2.28 Hz) 5.74 (1 H, dd, 7 =10.37, 2.28 Hz) 5.30-5.38 (0.6 H, m) 5.00-5.06 (0.4 H, m) 3.61-3.96 (8 H , m) 2.90-3.06 (1 H, m) 1.69-1.83 (1 H, m) 1.15-1.52 (6 H, m) 0.69-1.04 ( 6 H, m)

- 190 044387

10-1 503.1 Ή NMR (400 MHz, DMSO-t/ ₆ ) δ 10.15 (1 H, br s) 8.33 (1 Η, s) 7.36-7.45 (2 Η, m) 7.24-7 .36 (4 Η, m) 6.90 (1 Η, dd, 7=16.63, 10.37 Hz) 6.70-6.80 (2 N, m) 6.18 (1 H, dd, 7=16.73, 2.25 Hz) 5.75 (1 H, dd, 7=10.56, 2.15 Hz) 3.833.97 (4 N, m) 3.47-3.62 (4 H , m) 1.98-2.06 (3 H, m) 10-2 519.2 Ή NMR (CDC1 ₃ ) δ: 8.16-8.24 (m, 1H), 7.61-7.67 (m, 1H), 7.43-7.52 (m, 2H), 7.15 -7.23 (m, 1H), 7.05-7.13 (m, 1H), 6.92-7.02 (m, 1H), 6.70-6.82 (m, 2H), 6 .57-6.69 (m, 1H), 6.30-6.40 (m, 1H), 5.68-5.81 (m, 1H), 3.81-4.03 (m, 4H) , 3.49-3.71 (m, 7H), 2.52-2.66 (m, 1H). 10-3 537.0 Ή NMR (400 MHz, METHANOL-7 ₄ ) δ 8.35 (1 H, s) 7.387.42 (1 Η, m) 7.37 (1 Η, s) 7.31-7.36 (2 Η, m) 7.21 (1 Η, td, 7=8.31, 6.85 Hz) 6.83 (1 N, dd, 7=16.73, 10.66 Hz) 6.69 (1 H, d , 7=8.41 Hz) 6.63 (1 N, br t, 7=8.80 Hz) 6.24 (1 Η, dd, 7=16.82, 1.96 Hz) 5.77 (1 H, dd, 7=10.66, 1.86 Hz) 3.94-4.01 (4 H, m) 3.58-3.66 (4 H, m) 1.99-2.04 (3 H, m) 10-4 531.2 Ή NMR (400 MHz, METHANOL-7 ₄ ) δ 8.34 (1 H, s) 7.447.51 (2 H, t) 7.34 (1 H, d, 7 = 4.30 Hz) 7.27- 7.32 (1 N, w) 7.15-7.25 (2 N, w) 6.83 (1 N, dd, 7=16.82, 10.56 Hz) 6.66 (1 N, d , 7=8.22 Hz) 6.57-6.64 (1 N, w) 6.21-6.27 (1 N, w) 5.77 (1 N, dd, 7=10.66, 1 .86 Hz) 3.93-4.02 (4 N, t) 3.563.65 (4 N, t) 2.46-2.56 (1 N, t) 0.98-1.13 (6 N, T) 10-5 517.1 Ή NMR (400 MHz, METHANOL-7 ₄ ) δ 8.44 (1 H, s) 7.467.57 (3 H, t) 7.26-7.44 (3 H, t) 6.94 (1 H, dd, 7=16.73, 10.66 Hz) 6.77 (1 N, d, 7=8.22 Hz) 6.68-6.75 (1 N, t) 6.35 (1 N, dd , 7=16.73, 1.86 Hz) 5.88 (1 N, dd, 7=10.76, 1.96 Hz) 4.04-4.12 (4 N, t) 3.67-3 .76 (4 N, t) 2.36-2.60 (2 N, t) 1.06 (3 N, q, 7=7.63 Hz) 10-6 504.1 Ή NMR (400 MHz, METHANOL-7 ₄ ) δ 8.56 (1 H, br d, 7 = 5.09 Hz) 8.51 (1 H, s) 8.42 (1 H, s) 7.49 (1 N, br d, 7 = 5.09 Hz) 7.43 (1 N, s) 7.22-7.29 (1 N, t) 6.87 (1 N, dd, 7 = 16.73 , 10.66 Hz) 6.72 (1 N, br d, 7=8.41 Hz) 6.67 (1 N, brt, 7=8.71 Hz) 6.28 (1 N, dd, 7= 16.73, 1.27 Hz) 5.79-5.84 (1 N, t) 3.98-4.06 (4 N, t) 3.64-3.73 (4 N, t) 2, 19 (3 N, s)

- 191 044387

10-7 517.2 Ή NMR (400 MHz, METHANOL-7 ₄ ) δ 8.46 (1 H, s) 7.247.41 (5 H, m) 6.93 (1 H, dd, 7 = 16.82, 10.56 Hz) 6.76 (1 Η, d, 7=8.41 Hz) 6.68-6.74 (1 N, m) 6.34 (1 H, dd, 7=16.82, 1.96 Hz) 5 .87 (1 H, dd, 7=10.56, 1.96 Hz) 4.05-4.11 (4 Η, m) 3.68-3.76 (4 H, m) 2.02 (3 H, s) 2.00 (3 H, s) Ή NMR (400 MHz, METHANOL-7 ₄ ) δ 8.62 (1 H, dd, 7 = 4.89, 1.56 Hz) 8.43 (1 N, s) 7.87 (1 N, dd, 7=7.73, 1.47 Hz) 7.48 (1 N, dd, 7=7.63, 5.09 Hz) 7.43 (1 N , s) 7.27 (1 N, 10-8 504.2 td, 7=8.31, 6.85 Hz) 6.89 (1 N, dd, 7=16.82, 10.56 Hz) 6.74 (1 N, d, 7=8.22 Hz) 6 .68 (1 N, t, 7=8.80 Hz) 6.30 (1 N, dd, 7=16.73, 1.86 Hz) 5.83 (1 N, dd, 7=10.66, 1.86 Hz) 3.994.07 (4 H, t) 3.65-3.74 (4 H, t) 2.34 (3 H, s) Ή NMR (400 MHz, METHANOL-<7 ₄ ) δ 8 .33 (1 N, s) 7.75 (1 N, s) 7.52-7.56 (1 N, t) 7.12-7.28 (4 N, t) 6.82 (1 N, t) 10-9 528.2 dd, 7=16.73, 10.66 Hz) 6.64 (1 N, d, 7=8.41 Hz) 6.55-6.61 (1 N, t) 6.24 (1 N, dd , 7=16.82, 1.96 Hz) 6.14 (1 N, dd, 7=3.33, 0.78 Hz) 5.77 (1 N, dd, 7=10.56, 1.96 Hz) 3.914.01 (4 H, t) 3.55-3.63 (4 H, t) Ή NMR (400 MHz, METHANOL-7 ₄ ) δ 8.52 (1 H, s) 7.557.63 (2 N, t) 7.44-7.51 (2 N, t) 7.35-7.43 (1 N, t) 7.24 (1 N, t, 7=7.04 Hz) 7.02 ( 1 N, dd, 7=16.73, 10.66 Hz) 6.83- 10-10 529.1 6.90 (1 N, t) 6.76-6.83 (1 N, t) 6.43 (1 N, dd, 7=16.73, 1.66 Hz) 5.97 (1 N, dd , 7=10.66, 1.66 Hz) 4.13-4.19 (4 N, t) 3.76-3.82 (4 N, t) 1.56-1.70 (1 N, t ) 0.70-0.92 (3 H, t) 0.55-0.68 (1 H, t) Ή NMR (400 MHz, METHANOL-7 ₄ ) δ 8.40 (1 H, s) 7.507, 70 (4 N, t) 7.42 (1 N, s) 7.22-7.30 (1 N, t) 6.89 (1 N, 10-11 523.1 dd, 7=16.73, 10.66 Hz) 6.63-6.76 (2 N, t) 6.30 (1 N, dd, 7=16.73, 1.86 Hz) 5.83 ( 1 N, dd, 7=10.56, 1.96 Hz) 3.984.10 (4 N, t) 3.62-3.76 (4 N, t) Ή NMR (400 MHz, METHANOL-7 ₄ ) δ 8.45 (1 H, s) 7.637.67 (1 H, t) 7.51-7.54 (1 H, t) 7.44-7.48 (1 N, t) 7.34- 10-12 543.1 7.37 (1 N, t) 7.21 (1 N, td, 7=8.22, 6.85 Hz) 6.90 (1 N, dd, 7=16.73, 10.66 Hz) 6 .58-6.72 (2 N, t) 6.31 (1 N, dd, 7=16.82, 1.96 Hz) 5.84 (1 N, dd, 7=10.56, 1.96 Hz) 4.03-

- 192 044387

4.08 (4 H, m) 3.69-3.75 (4 Η, m) 2.22 (1.25 Η, s) 2.20 (1.75 Η, s) 10-13 447.0 Ή NMR (400 MHz, DMSO-t/ ₆ ) δ 10.26 (1 Η, br s) 8.31 (1 Η, s) 8.14 (1 Η, s) 7.31-7.40 (1 Η, m) 6.78-6.92 (3 Η, m) 6.17 (1 Η, dd, 7=16.63, 2.35 Hz) 5.74 (1 N, dd, 7=10, 37, 2.35 Hz) 3.79-3.92 (4 H, m) 3.46-3.55 (4 Η, m) 11-1-1 567.2 Ή NMR (400 MHz, CDC1 ₃ ) δ 10.28 (1 Η, br s) 7.94 (1 Η, s) 7.35-7.49 (4 Η, m) 7.25-7.31 ( 2 Η, m) 7.11 (1 Η, d, 7=7.67 Hz) 6.64 (1 N, dd, 7=16.79, 10.57 Hz) 6.54 (1 H, s) 6.41 (1 Η, dd, 7=16.79, 1.87 Hz) 5.81 (1 N, dd, 7=10.57, 1.66 Hz) 3.83-4.07 (8 H , m) 2.74 (1 Η, spt, 7=6.84 Hz) 2.13 (3 H, s) 1.23 (3 H, d, 7=6.84 Hz) 1.04 (3 H , d, 7=6.84 Hz) 11-1-2 567.2 Ή NMR (400 MHz, CDC1 ₃ ) δ 10.37 (1 H, br s) 7.94 (1 Η, s) 7.34-7.50 (4 H, m) 7.21-7.31 ( 2 H, m) 7.13 (1 H, d, 7=7.67 Hz) 6.64 (1 H, dd, 7=16.90, 10.68 Hz) 6.55 (1 H, s) 6.41 (1 H, dd, 7=16.79, 1.66 Hz) 5.81 (1 H, dd, 7=10.47, 1.55 Hz) 3.83-4.08 (8 N , m) 2.70 (1 H, spt, 7=6.84 Hz) 2.13 (3 H, s) 1.22 (3 Η, d, 7=6.84 Hz) 1.03 (3 N , d, 7=6.84 Hz). MS (ESI, +ve) mass/charge: 567.2 [M+H] ⁺ . 11-2-1 581.3 NMR (400 MHz, METAHOJI-d ₄ ) δ 8.12-8.21 (m, 1H), 7.30-7.56 (m, 6H), 7.21 (d, 7=7.7 Hz, 1H), 6.80-6.97 (m, 1H), 6.46 (s, 1H), 6.30-6.41 (m, 1H), 5.79-5.94 (m, 1H) , 5.02-5.14 (m, 1H), 4.39-4.69 (m, 2H), 4.07-4.30 (m, 1H), 3.67 (s, 2H), 3 ,21-3.51 (m, 1H), 2.68-2.84 (m, 1H), 2.13 (s, 3H), 1.54 (br d, 7=6.0 Hz, 3H) , 1.23 (d, 7=7.1 Hz, 3H), 1.05 (d, 7=6.8 Hz, 3H) 11-2-2 581.2 Ή NMR (400 MHz, METHANOL) δ 8.15 (s, 1H), 7.427.58 (m, 4H), 7.30-7.39 (m, 2H), 7.25 (d, 7=7, 7 Hz, 1H), 6.81-6.98 (m, 1H), 6.48 (s, 1H), 6.29-6.41 (m, 1H), 5.87 (dd, 7=1 ,35, 10.68 Hz, 1H), 5.02-5.12 (m, 1H), 4.42-4.69 (m, 2H), 4.05-4.29 (m, 1H), 3.65-3.93 (m, 2H), 3.21-3.47 (m, 1H), 2.64-2.79 (m, 1H), 2.15 (s, 3H), 1, 53 (br d, 7=6.6 Hz, 3H), 1.23 (br d, 7=6.8 Hz, 3H), 1.06 (d, 7=6.8 Hz, 3H)

- 193 044387

Table 14(a)

Analytical data for selected examples

Note No. LRMS: (ESI, +ve ion) mass/charge NMR 12 453.2 Ή NMR (CDC1 ₃ ) δ: 8.30-8.37 (m, 1H), 8.11-8.18 (m, 1H), 7.29-7.38 (m, 1H), 6.96 -7.18 (m, 1H), 6.88-6.94 (m, 1H), 6.76-6.85 (m, 1H), 6.59-6.72 (m, 1H), 6 .31-6.42 (m, 1H), 5.73-5.84 (m, 1H), 3.73-4.05 (m, 4H), 3.35-3.62 (m, 4H) , 2.40-2.52 (m, 1H), 1.35-1.42 (m, 1H), 1.29-1.34 (m, 1H), 1.03-1.14 (m, 2H). mass/charge (ESI) M+H: 453.2. 13 504.2 NMR (CDC1 ₃ ) δ: 7.96-8.09 (m, 2H), 7.46-7.57 (m, 2H), 7.37-7.44 (m, 1H), 7.29- 7.33 (m, 1H), 7.20-7.26 (m, 1H), 6.96-7.07 (m, 1H), 6.81-6.87 (m, 1H), 6. 70-6.77 (m, 1H), 6.54-6.67 (m, 1H), 6.29-6.41 (m, 1H), 5.68-5.82 (m, 1H), 3.74-3.96 (m, 4H), 3.12-3.43 (m, 4H). 14 481.2. NMR (CDC1 ₃ ) δ: 8.10-8.22 (m, 2H), 7.29-7.38 (m, 1H), 6.86-6.93 (m, 1H), 6.77- 6.85 (m, 1H), 6.61-6.72 (m, 1H), 6.33-6.44 (m, 1H), 5.74-5.85 (m, 1H), 3, 82-4.05 (m, 4H), 3.75-3.82 (m, 1H), 3.40-3.63 (m, 4H), 2.06-2.24 (m, 4H), 1.81-1.96 (m, 2H), 1.67-1.79 (m, 2H). 15 496.2 NMR (CDC1 ₃ ) δ: 8.08-8.15 (m, 1H), 7.98-8.05 (m, 1H), 7.29-7.39 (m, 1H), 6.86- 6.94 (m, 1H), 6.76-6.85 (m, 1H), 6.59-6.70 (m, 1H), 6.30-6.43 (m, 1H), 5, 72-5.84 (m, 1H), 3.77-4.05 (m, 4H), 3.40-3.56 (m, 4H), 3.32-3.38 (m, 4H), 1.73-1.85 (m, 4H), 1.64-1.70 (m, 2H) 16 505.2 NMR (CDC1 ₃ ) δ: 8.41-8.45 (m, 1H), 8.17-8.20 (m, 1H), 7.40-7.45 (m, 2H), 7.28- 7.37 (m, 2H), 7.20-7.26 (m, 1H), 6.78-6.87 (m, 2H), 6.59-6.70 (m, 1H), 6. 31-6.41 (m, 1H), 5.97-6.06 (m, 1H), 5.74-5.81 (m, 1H), 3.76-4.03 (m, 4H), 3.38-3.53 (m, 4H). 17-1 539.2 NMR (400 MHz, DMSO-A) δ 9.97 (s, 1H), 8.10 (s, 1H), 7.80 (d, 7=8.4 Hz, 1H), 7.40-7, 46 (m, 1H), 7.19-7.30 (m, 3H), 7.97 (d, 7=2.4 Hz, 1H), 6.62-6.71 (m, 2H), 3 .80-3.93 (m, 4H), 3.62-3.69 (m, 4H), 3.07 (d, 7=4.1 Hz, 2H), 2.17 (s, 6H).

- 194 044387

17-2 525.0 Ή NMR (400 MHz, DMSO-t/ ₆ ) δ 9.97 (s, 1H), 8.10 (s, 1H), 7.80 (d, 7=8.4 Hz, 1H), 7.40 -7.46 (m, 1H), 7.19-7.30 (m, 3H), 7.97 (d, 7=2.4 Hz, 1H), 6.62-6.71 (m, 2H ), 3.80-3.93 (m, 4H), 3.62-3.69 (m, 4H), 3.07 (d, 7=4.1 Hz, 2H), 2.17 (s, 6H). 18-1 512.0 Ή NMR (400 MHz, DMSO-7 ₆ ) δ 8.12 (s, 1H), 7.94 (d, 7=8.2 Hz, 1H), 7.47-7.55 (m, 2H), 7.25-7.34 (m, 2H), 7.19 (d, 7=2.5 Hz, 1H), 5.43 (br. s, 1H), 5.20 (br. s, 1H) , 5.14 (t, 7=5.8 Hz, 1H), 4.12 (d, 7=5.7 Hz, 2H), 3.94 (s, 3H), 3.783.85 (m, 4H) , 3.54-3.66 (m, 4H). 18-2 560.0 Ή NMR (400 MHz, DMSO-7 ₆ ) δ 9.96 (br. s, 1H), 8.13 (s, 1H), 7.80 (d, 7=8.2 Hz, 1H), 7, 40-7.47 (m, 1H), 7.19-7.29 (m, 3H), 7.07 (d, 7=2.4 Hz, 1H), 5.41 (s, 1H), 4 .38 (s, 1H), 4.38 (s, 2H), 3.84-3.93 (m, 4H), 3.62-3.72 (m, 4H). 18-3 498.0 Ή NMR (400 MHz, DMSO-7 ₆ ) δ 9.98 (br. s, 1H), 8.11 (s, 1H), 7.79 (d, 7=8.2 Hz, 1H), 7, 37-7.48 (m, 1H), 7.17-7.28 (m, 3H), 7.07 (d, 7=2.4 Hz, 1H), 5.43 (br. s, 1H) , 5.20 (br. s, 1H), 5.07-5.14 (m, 1H), 4.12 (br. s, 2H), 3.78-3.86 (m, 4H), 3 .57-3.66 (m, 4H). 19-1 486.0 NMR (400 MHz, DMSO-7 ₆ ) δ 8.13 (s, 1H), 8.02 (s, 1H), 7.33 (d, 7=2.2 Hz, 1H), 6.99 (d , 7=2.4 Hz, 1H), 6.85 (dd, 7=16.6, 10.6 Hz, 1H), 6.18 (dd, 7=16.7, 2.3 Hz, 1H) , 5.76 (dd, 7=10.5, 2.3 Hz, 1H), 3.85-3.95 (m, 4H), 3.84 (s, 3H), 3.62-3.72 (m, 4H), 3.56 (s, 3H). 19-2 472.0 NMR (400 MHz, DMSO-7 ₆ ) δ 9.40 (s, 1H), 8.12 (s, 1H), 7.92 (s, 1H), 7.12 (d, 7=2.2 Hz , 1H), 6.81-6.91 (m, 2H), 6.18 (dd, 7=16.7, 2.5 Hz, 1H), 5.76 (dd, 7=10.4, 2 .4 Hz, 1H), 3.81-3.94 (m, 4H), 3.62-3.70 (m, 4H), 3.52 (s, 3H). 19-3 472.0 Ή NMR (400 MHz, DMSO-7 ₆ ) δ 9.28 (s, 1H), 8.11 (s, 1H), 8.01 (s, 1H), 6.95 (d, 7=2.0 Hz, 1H), 6.77-6.90 (m, 2H), 6.18 (dd, 7=16.7, 2.5 Hz, 1H), 5.76 (dd, 7=10.4, 2.2 Hz, 1H), 4.03 (s, 3H), 3.80-3.94 (m, 4H), 3.58-3.66 (m, 4H). 20 512 Ή NMR (400 MHz, DMSO-7 ₆ ) δ 9.93 (br s, 1H), 8.11 (s, 1H), 7.80 (d, 7=12 Hz, 1H), 7.43 (m , 1H), 7.26-7.20 (m,

- 195 044387

ZH), 7.07 (s, 1H), 5.32 (s, 1H), 5.16 (s, 1H), 3.83 (br s, 4H), 3.63 (br s, 4H), 3.53 (t, /=8.0 Hz, 2H), 2.42 (t, /=8.0 Hz, 2H). ¹⁹ NMR (377 MHz, DMSO-/,) δ -123.8 (s, IF). 21 469 Ή NMR (400 MHz, CDC1 ₃ ) δ 8.81 (dd, /=4.2, 1.3 Hz, 1 Η) 7.72-7.78 (m, 2 Η) 7.64 (s, 1 Η) 7.28 (d, /=2.2 Hz, 1 Η) 7.16 (dd, /=8.4, 4.3 Hz, 1 Η) 6.56-6.66 (m, 1 Η ) 6.40 (dd, /=16.8, 1.6 Hz, 1 Η) 5.78-5.87 (m, 1 Η) 4.01 (br. s, 2 Η) 3.89 (br s, 2 H) 3.50-3.60 (m, 4 H). ¹⁹ F NMR (376 MHz, CDC1 ₃ ) δ-121.33 (s, 1 F). 22 512.0 Ή NMR (400 MHz, DMSO-/,) δ 3.14-3.28 (m, 1 H) 3.523.87 (m, 3 H) 4.15-5.03 (m, 2 H) 5.15 -5.23 (m, 1 H) 5,775.83 (m, 1 H) 6.13-6.24 (m, 1 H) 6.86 (br. s, 1 H) 7,067.12 (m, 1 H) 7.20-7.30 (m, 3 H) 7.38-7.49 (m, 1 H) 7.767.84 (m, 1 H) 8.07-8.13 (m, 1 H) 9.98 (br. s, 1 H) 13.42 (br. s, 1 H). 23 482.0 Ή NMR (400 MHz, DMSO-de) δ 12.92-13.19 (1 H, m), 8.02-8.21 (1 H, m), 7.47-7.60 (2 H, m), 7.02-7.09 (1 H, m), 6.80-6.93 (1 H, m), 6.15-6.25 (1 H, m), 5.71-5 .82 (1 H, m), 3.80-3.96 (4 H, m), 3.60-3.72 (4 H, m), 1.55-1.74 (1 H, m) , 0.72-0.79 (2 H, m), 0.58-0.71 (2 H, m). 24 482.0 Ή NMR (400 MHz, METAHOJI-d ₄ ) δ 7.90 (s, 1H), 7.63 (d, /=8.5 Hz, 1H), 7.43 (t, /=7.4 Hz, 1H), 7.24 (d, /=8.5 Hz, 1H), 7.05 (t, /=7.4 Hz, 1H), 6.72-6.84 (m, 1H), 6, 67 (s, 1H), 6.15-6.28 (m, 1H), 5.68-5.81 (m, 1H), 3.87-3.97 (m, 4H), 3.63 ( m, 4H), 2.90 (s, 3H). 25 468.0 Ή NMR (400 MHz, METAHOJI-d ₄ ) δ 7.85 (s, 1H), 7.53 (d, /=8.5 Hz, 1H), 7.39 (t, /=7.6 Hz, 1H), 7.23 (d, /=8.5 Hz, 1H), 7.03 (t, /=7.8 Hz, 1H), 6.82 (s, 1H), 6.71 (dd, /=10.8, 16.8 Hz, 1H), 6.2 (dd, /=1.5, 16.8 Hz, 1H), 5.70 (dd, /=1.5, 10.8 Hz , 1H), 3.82-3.93 (m, 4H), 3.50-3.66 (m, 4H) 26 468.0 Ή NMR (400 MHz, METAHOJI-d ₄ ) δ 7.90 (s, 1H), 7.53 (d, /=8.2 Hz, 1H), 7.42-7.49 (m, 1H), 7.10 (d, /=8.0 Hz, 1H), 7.03-7.08 (m, /=7.6 Hz, 1H), 6.67-6.81 (m, 2H), 6 .19 (dd, /=1.8, 16.6 Hz, 1H), 5.72 (dd, /=1.8, 10.6 Hz, 1H),

- 196 044387

3.87-3.93 (m, 4H), 3.56-3.66 (m, 4H). 27 512.3 Ή NMR (400 MHz, CDC1 ₃ ) δ 9.51 (0.6 Η, br s) 8.98 (0.4 Η, br s) 7.63 (0.4 H, s) 7.58 (0 ,6 H, s) 7.35-7.43 (2 H, m) 7.10-7.26 (3 H, m) 6.78 (1 H, dd, 7=16.63, 8.22 Hz) 6.596.71 (2 H, m) 6.36 (1 Η, dd, 7=16.82, 1.57 Hz) 5.78 (1 Η, dd, 7=10.56, 1.37 Hz ) 4.10-4.38 (4 H, m) 3.80-4.03 (4 Η, m) 2.60-2.72 (1 Η, m) 2.61 (1.2 Η, s ) 2.59 (1.8 Η, s) 0.911.08 (6 Η, m) 28 517.1 Ή NMR (400 MHz, METHANOL-7 ₄ ) δ 8.27 (1 H, s) 8.15 (0.33 H, s) 8.10 (0.67 H, s) 7.19-7.31 (5 H, m) 7.10-7.16 (1 H, m) 6.86 (1 H, dd, 7=16.73, 10.66 Hz) 6.62-6.78 (2 Η, m) 6.27 (1 Η, dd, 7=16.82, 1.96 Hz) 5.80 (1 N, dd, 7=10.66, 1.86 Hz) 4.94-5.01 ( 1 H, m) 3.93-4.03 (4 Η, m) 3.49-3.60 (4 Η, m) 1.81 (3 Η, d, 7=7.04 Hz) 29 521.1 Ή NMR (400 MHz, METHANOL-7 ₄ ) δ 8.32 (1 H, s) 8.19 (1 H, s) 7.26-7.34 (3 H, w) 6.98 (2 H, t, 7=8.71 Hz) 6.696.91 (3 N, w) 6.28 (1 N, dd, 7=16.92, 1.86 Hz) 5.82 (1 N, dd, 7=10 .56, 1.76 Hz) 4.54-4.65 (2 N, t) 3.99 (4 N, t) 3.58 (4 N, t) thirty 489.0 Ή NMR (400 MHz, CHLOROFORM-7) δ 8.21 (1 H, s) 8.06 (1 H, s) 7.62-7.69 (2 H, t) 7.45-7.51 ( 3 N, t) 7.24-7.32 (1 N, t) 6.81-6.90 (1 N, t) 6.75 (1 N, t, 7=8.41 Hz) 6.65 (1 N, dd, 7 = 16.82, 10.56 Hz) 6.38 (1 N, dd, 7 = 16.82, 1.76 Hz) 5.79 (1 N, dd, 7 = 10, 56, 1.76 Hz) 3.86-4.02 (4 N, t) 3.57-3.76 (4 N, t) 31 489.1 Ή NMR (400 MHz, CDC1 ₃ ) δ 8.15 (1 H, s) 8.12 (1 H, s) 7.68-7.73 (2 H, m) 7.53-7.58 (3 H, m) 7.30 (1 H, br td, 7=8.22, 6.65 Hz) 6.88 (1 H, d, 7=8.22 Hz) 6.78 (1 H, t, 7=8.61 Hz) 6.57 (1 N, dd, 7=16.82, 10.56 Hz) 6.28 (1 Η, dd, 7=16.73, 1.66 Hz) 5.71 (1 N, dd, 7=10.56, 1.56 Hz) 3.78-3.89 (4 H, t) 3.51-3.73 (4 Η, t)

- 197044387

32 443.1 Ή NMR (400 MHz, CDC1 ₃ ) δ 8.23 (1 Η, s) 8.11 (1 Η, s) 7.32 (1 Η, td, /=8.31, 6.46 Hz) 6, 88 (1 Η, d, /=8.22 Hz) 6.77-6.83 (1 N, m) 6.65 (1 H, dd, /=16.82, 10.56 Hz) 6.37 (1 H, dd, /=16.82, 1.76 Hz) 5.79 (1 H, dd, /=10.47, 1.86 Hz) 4.18 (3 H, s) 3.79- 4.05 (4 Η, m) 3.34-3.54 (4 Η, m) 33 443.1 Ή NMR (400 MHz, CDC1 ₃ ) δ 8.32 (1 Η, s) 8.01 (1 Η, s) 7.32 (1 Η, td, /=8.27, 6.55 Hz) 6, 89 (1 Η, d, /=8.22 Hz) 6.77-6.83 (1 N, m) 6.60 (1 H, dd, /=17.02, 10.56 Hz) 6.30 (1 H, dd, /=16.82, 1.76 Hz) 5.75 (1 H, dd, /=10.56, 1.76 Hz) 4.22 (3 H, s) 3.67- 3.98 (4 Η, m) 3.25-3.55 (4 Η, m) 34 427.1 Ή NMR (400 MHz, CDC1 ₃ ) δ ppm 8.13 (1 Η, s) 8.12 (1 Η, s) 7.28-7.36 (4 Η, m) 7.20-7.26 ( 1 Η, m) 6.65 (1 Η, dd, /=16.82, 10.56 Hz) 6.37 (1 N, dd, /=16.82, 1.57 Hz) 5.78 (1 H, dd, /=10.56, 1.56 Hz) 4.61 (2 H, s) 3.83-4.01 (4 Η, m) 3.48-3.62 (4 H, m) . 35 503.1 Ή NMR (400 MHz, CDC1 ₃ ) δ 8.13 (1 H, s) 8.11 (1 Η, s) 7.12-7.37 (6 Η, m) 6.91 (1 Η, d, /=8.22 Hz) 6.77 (1 H, t, /=8.61 Hz) 6.64 (1 H, dd, /=16.82, 10.56 Hz) 6.37 (1 Η, dd, /=16.82, 1.76 Hz) 5.79 (1 H, dd, /=10.56, 1.96 Hz) 4.55 (2 H, s) 3.34-4.01 ( 8 H, m) 36 503.1 Ή NMR (400 MHz, CDC1 ₃ ) δ 8.12 (1 H, s) 8.05 (1 Η, s) 7.26-7.36 (5 Η, m) 7.19-7.24 (1 Η, m) 6.93 (1 Η, d, /=8.41 Hz) 6.76 (1 H, t, /=8.31 Hz) 6.58 (1 H, dd, /=16.82 , 10.76 Hz) 6.28 (1 H, dd, /=16.82, 1.76 Hz) 5.75 (1 H, dd, /=10.56, 1.76 Hz) 4.54 ( 2 N, s) 3.32-3.93 (8 H, m) 37 523 Ή NMR (400 MHz, CDC1 ₃ ) δ 8.23 (0.6 H, s) 8.22 (0.4 Η, s) 8.02 (0.4 Η, s) 8.00 (0.6 Η, s) 7.19-7.57 (8 Η, m) 6.68 (0.4 Η, dd, /=16.82, 10.56 Hz) 6.60 (0.6 N, dd, /=16.82, 10.56 Hz) 6.38 (0.4 H, dd, /=16.63, 1.76 Hz) 6.32 (0.6 H, dd, /=16.82, 1.76 Hz) 5.79 (0.4 H, dd, /=10.56, 1.76 Hz) 5.73 (0.6 H, dd, /=10.56, 1.76 Hz) 4 .67 (1.2 H, s) 4.60 (0.8 Η, s) 3.74-4.06 (4 Η, m) 3.46-3.70 (4 Η, m) 2.21 (1.8 Η, s) 2.06 (1.2 Η, s) 38 483.3 Ή NMR (400 MHz, CDC1 ₃ ) δ 8.04 (1 Η, s) 7.26-7.33 (1 Η, m) 6.82 (1 Η, d, /=8.29 Hz) 6, 71 (1 Η, t, /=8.91 Hz) 6.51 (1 H, dd, /=16.79, 10.57 Hz) 6.30 (1 H, dd, /=16.79, 1 .45 Hz) 5.72 (1 H, dd, /=10.47, 1.55 Hz) 4.15 (2 H, br d, /=6.43 Hz) 3.69-3.90 (8 H, m) 1.14-1.27 (4 Η, m) 0.73-0.88 (1 Η, m)

- 198044387

Table 14(b)

Analytical data for selected examples__________

Note No. LRMS: (ESI, +ve ion) mass/charge NMR 39-1 576.0 Ή NMR (400 MHz, DMSO-7 ₆ ) δ ppm 10.07 (s, 1 Η), 9.9610.13 (m, 1 Η), 8.39 (br s, 1H), 7.17-7 .26 (m, 3 H), 7.09 (br d, 7=5.9 Hz, 1 H), 6.84 (br s, 1 H), 6.60-6.71 (m, 2 H ), 6.20 (br d, 7=17.2 Hz, 1 H), 5.74-5.78 (m, 1 H), 4.77-4.99 (m, 1 H), 4, 26 (br s, 3 H), 3.58-3.86 (m, 1 H), 3.57 (s, 1 H), 3.13 (brd, 7=1.7 Hz, 1 H), 1.85 (br s, 3 H), 1.34 (br d, 7=6.4 Hz, 3 H), 1.05 (d, 7=6.6 Hz, 3 H), 0.92 ( d, 7=6.6 Hz, 3 H). ¹⁹ F NMR (376 MHz, CHLOROFORM 7) δ ppm -104.54 (br d, 7=10.4 Hz, IF). 39-2 576.0 ^X H NMR (400 MHz, DMSO-7 ₆ ) δ ppm 10.07 (s, 1 H), 8.42 (br s, 1 H), 7.19-7.26 (m, 3 H), 7 ,10 (br d, 7=6.0 Hz, 1 H), 6.80-6.91 (m, 1 H), 6.70 (d, 7=8.0 Hz, 1 H), 6, 65 (t, 7=8.7 Hz, 1 H), 6.21 (br d, 7=15.8 Hz, 1 H), 5.74-5.79 (m, 1 H), 4.94 (br s, 1 H), 4.27 (br s, 2 H), 4.03 (br d, 7=12.4 Hz, 1 H), 3.61-3.86 (m, 2 H) , 2.94-3.20 (m, 1 H), 1.85 (br s, 3 H), 1.32 (br d, 7 = 6.2 Hz, 3 H), 1.02-1, 08 (m, 4 H), 0.92 (d, 7 = 6.8 Hz, 3 H). ¹⁹ F NMR (376 MHz, CHLOROFORM 7) δ ppm -104.83 - -103.96 (m, IF). 40-1 560.9 Ή NMR (400 MHz, DMSO-7 ₆ ) δ ppm 8.45 (br d, 7 = 5.4 Hz, 1 H), 8.39 (d, 7 = 5.0 Hz, 1 H), 7, 47-7.55 (m, 1 H), 7,177.34 (m, 4 H), 6.81-6.92 (m, 1 H), 6.16-6.26 (m, 1 H), 5.73-5.80 (m, 1 H), 4.95 (br s, 1 H), 3.99-4.45 (m, 3 H), 3.71-3.85 (m, 1 H), 3.60-3.69 (m, 0.5 H), 3.41-3.53 (m, 0.5 H), 3.06-3.27 (m, 1 H), 2 .65-2.75 (m, 1 H), 1.94 (s, 3 H), 1.34 (d, 7=6.6 Hz, 3 H), 1.07 (d, 7=6, 6 Hz, 3 H), 0.94 (d, 7=6.6 Hz, 3 H).

- 199 044387

40-2 560.9 Ή NMR (400 MHz, DMSO-J ₆ ) δ ppm 8.46 (br d, 7=4.6 Hz, 1 Η), 8.39 (d, 7=4.8 Hz, 1 Η),7, 47-7.55 (m, 1 Η), 7.177.34 (m, 4 Η), 6.81-6.92 (m, 1 Η), 6.16-6.26 (m, 1 Η), 5.73-5.80 (m, 1 Η), 4.95 (br s, 1 Η), 3.99-4.45 (m, 3 Η), 3.71-3.85 (m, 1 Η), 3.60-3.69 (m, 0.5 Η), 3.41-3.53 (m, 0.5 Η), 3.06-3.27 (m, 1 Η), 2 .65-2.75 (m, 1 Η), 1.93 (s, 3 Η), 1.34 (d, 7=6.6 Hz, 3 Η), 1.07 (d, 7=6, 6 Hz, 3 Η), 0.94 (d, 7 = 6.6 Hz, 3 Η). 41-1 561.2 Ή NMR (400 MHz, DMSO-J ₆ ) δ ppm 10.20 (s, 1 H), 8.39 (d, 7 = 4.8 Hz, 1 H), 8.24-8.34 (m, 1 H), 7.23-7.32 (m, 1 H), 7.19 (d, 7=5.0 Hz, 1 H), 6.87 (td, 7=16.3, 11.0 Hz, 1 H), 6.74 (d, 7=8.6 Hz, 1 H), 6.69 (t, 7=8.6 Hz, 1 H), 6.21 (br d, 7=16 .2 Hz, 1 H), 5.74-5.80 (m, 1 H), 4.91 (br s, 1 H), 4.234.45 (m, 2 H), 3.97-4.21 (m, 1 H), 3.44-3.79 (m, 2 H), 3.11-3.31 (m, 1 H), 2.67-2.77 (m, 1 H), 1 .91 (s, 3 H), 1.35 (d, 7=6.8 Hz, 3 H), 1.08 (d, 7=6.6 Hz, 3 H), 0.94 (d, 7 =6.8 Hz, 3 H). 41-2 561.2 Ή NMR (400 MHz, DMSO-J ₅ ) δ ppm 10.20 (s, 1 H), 8.39 (d, 7 = 4.8 Hz, 1 H), 8.24-8.34 (m, 1 H), 7.23-7.32 (m, 1 H), 7.19 (d, 7=5.0 Hz, 1 H), 6.87 (td, 7=16.3, 11.0 Hz, 1 H), 6.74 (d, 7=8.6 Hz, 1 H), 6.69 (t, 7=8.6 Hz, 1 H), 6.21 (br d, 7=16 .2 Hz, 1 H), 5.74-5.80 (m, 1 H), 4.91-4.99 (br s, 1 H), 4.23-4.45 (m, 2 H) , 3.97-4.21 (m, 1 H), 3.44-3.79 (m, 2 H), 3.11-3.31 (m, 1 H), 2.67-2.77 (m, 1 H), 1.91 (s, 3 H), 1.35 (d, 7=6.8 Hz, 3 H), 1.08 (d, 7=6.6 Hz, 3 H) , 0.94 (d, 7=6.8 Hz, 3 H). 42-1 559.0 Ή NMR (400 MHz, DMSOT) δ ppm 10.25 (br s, 1 H), 8.16-8.34 (m, 2 H), 7.22-7.34 (m, 1 H), 7 .10 (d, 7=5.0 Hz, 1 H), 6.79-6.94 (m, 1 H), 6.58-6.77 (m, 2 H), 6.13-6, 27 (m, 1 H), 5.70-5.83 (m, 1 H), 4.86 (br s, 1 H), 4.26-4.48 (m, 2 H), 3.95 -4.20 (m, 1 H), 3.45-3.73 (m, 2 H), 3.113.27 (m, 1 H), 1.91 (s, 3 H), 1.68 (br d, 7=3.9 Hz, 1 H), 1.36 (d, 7=6.6 Hz, 3 H), 0.82-0.88 (m, 1 H), 0.71-0, 82 (m, 2 H), 0.58-0.66 (m, 1 H). ¹⁹ F NMR (376 MHz, DMSO-J ₆ ) δ ppm -116.15 (s, 1 F), -127.30 (s, IF).

- 200 044387

42-2 559.0 Ή NMR (400 MHz, DMSO-t/ ₆ ) δ ppm 10.22 (s, 1 Η), 8.288.35 (m, 1 Η), 8.25 (br d, 7=4.4 Hz, 1 Η ), 7.28 (q, 7=7.6 Hz, 1 Η), 7.11 (br d, 7=4.4 Hz, 1 Η), 6.80-6.93 (m, 1 Η) , 6.656.78 (m, 2 Η), 6.20 (br d, 7=16.8 Hz, 1 Η), 5.76 (br d, 7=10.6 Hz, 1 Η), 4.96 (br d, 7=3.1 Hz, 1 Η), 4.20-4.46 (m, 2 Η), 3.97-4.18 (m, 1 Η), 3.39-3.83 (m, 2 Η), 3.02-3.29 (m, 1 Η), 1.92 (s, 3 Η), 1.69 (br s, 1 Η), 1.31 (br d, 7 =6.0 Hz, 3 Η), 0.71-0.90 (m, 3 Η), 0.62 (br d, 7 = 6.4 Hz, 1 H). ¹⁹ F NMR (376 MHz, DMSO-7 ₆ ) δ ppm -115.37 (s, 1 F), -128.36 (s, IF). 43 590.2 Ή NMR (400 MHz, DMSO-7 ₆ ) δ ppm 9.06 (s, 1 H), 8.468.52 (m, 1 H), 7.48-7.55 (m, 1 H), 7.26 -7.34 (m, 2 H), 7.17 (td, 7=7.4, 1.6 Hz, 1 H), 6.82-6.93 (m, 1 H), 6.22 ( br d, 7=16.6 Hz, 1H), 5.75-5.80 (m, 1 H), 5.00 (brs, 1 H), 4.314.43 (m, 2 H), 4.02 -4.21 (m, 1 H), 3.81 (br d, 7=8.9 Hz, 1 H), 3.45-3.70 (m, 1 H), 3.10-3.30 (m, 1 H), 2.73 (br d, 7=6.4 Hz, 2 H), 1.36 (d, 7=6.6 Hz, 3 H), 1.09 (d, 7= 6.6 Hz, 6 H), 0.93 (d, 7=6.4 Hz, 6 H). 44-1 574.9 Ή NMR (400 MHz, DMSO-J ₆ ) δ 10.07-10.22 (m, 1 H), 8.39 (br s, 1 H), 8.24 (d, 7 = 4.8 Hz, 1 H), 7.25 (q, 7=8.1 Hz, 1 H), 7.12 (br s, 1 H), 6.79-6.93 (m, 1 H), 6.62- 6.75 (m, 2 H), 6.20 (br d, 7=15.8 Hz, 1 H), 5.71-5.83 (m, 1 H), 4.88 (br s, 1 H), 4.25-4.45 (m, 2 H), 3.98-4.21 (m, 1 H), 3.54 (br d, 7=5.8 Hz, 2 H), 3 .09-3.32 (m, 1 H), 1.92 (brd, 7=15.8 Hz, 3 H), 1.62-1.82 (m, 1 H), 1.37 (br s , 3 H), 0.85 (br s, 1 H), 0.76 (br s, 2 H), 0.65 (br s, 1 H). 44-2 574.9 Ή NMR (400 MHz, DMSO-7 ₆ ) δ 10.07-10.21 (m, 1 H), 8.44 (br s, 1 H), 8.25 (d, 7 = 5.0 Hz, 1 H), 7.25 (q, 7=7.9 Hz, 1 H), 7.13 (br s, 1 H), 6.86 (dt, 7=16.9, 8.3 Hz, 1 H), 6.636.75 (m, 2 H), 6.21 (br d, 7=16.8 Hz, 1 H), 5.72-5.80 (m, 1 H), 4.99 (br d, 7=2.3 Hz, 1 H), 4.21-4.46 (m, 2 H), 4.014.18 (m, 1 H), 3.40-3.59 (m, 2 H) , 3.02-3.34 (m, 1 H), 1.92 (br d, 7=18.5 Hz, 3H), 1.62-1.81 (m, 1 H), 1.31 ( br s, 3 H), 0.83-0.91 (m, 1 H), 0.77 (br s, 2 H), 0.65 (br s, 1 H).

- 201 044387

45-1 559.0 Ή NMR (400 MHz, DMSO-/ ₆ ) δ ppm 8.44 (br s, 1 H), 8.25 (d, /=5.0 Hz, 1 H), 7.49-7.57 (m , 1 H), 7.30-7.35 (m, 1 H), 7.24-7.30 (m, 2 H), 7.11 (d, /=5.0 Hz, 1 H), 6.80-6.93 (m, 1 H), 6.15-6.25 (m, 1 H), 5.73-5.80 (m, 1 H), 4.94 (br d, / =4.6 Hz, 1 H), 4.28 (br s, 1 H), 3.98-4.19 (m, 1 H), 3.703.82 (m, 2 H), 3.41-3 .68 (m, 1 H), 3.05-3.29 (m, 1 H), 1.97 (s, 3 H), 1.64 (br d, /=4.1 Hz, 1 H) , 1.34 (d, /=6.8 Hz, 3 H), 0.86-0.94 (m, 1 H), 0.77-0.84 (m, 1 H), 0.70- 0.76 (m, 1 H), 0.67 (br d, /=8.1 Hz, 1 H). ¹⁹ F NMR (376 MHz, DMSO/ ₆ ) δ ppm-113.35 (s, IF). 45-2 559.0 Ή NMR (400 MHz, DMSOR) δ ppm 8.44 (br d, /=4.6 Hz, 1 H), 8.25 (d, /=4.8 Hz, 1 H), 7.48-7 .57 (m, 1 H), 7.257.35 (m, 3 H), 7.12 (d, /=5.0 Hz, 1 H), 6.77-6.94 (m, 1 H), 6.20 (br d, /=17.2 Hz, 1 H), 5.71-5.79 (m, 1 H), 4.95 (br s, 1 H), 4.24-4.44 (m, 2 H), 3.98-4.22 (m, 1 H), 3.71-3.87 (m, 1 H), 3.56-3.67 (m, 1 H), 3 .20-3.30 (m, 1 H), 1.96 (s, 3 H), 1.68 (br s, 1 H), 1.34 (d, /=6.6 Hz, 3 H) , 0.86-0.93 (m, 1 H), 0.73-0.84 (m, 2 H), 0.61-0.72 (m, 1 H). ¹⁹ F NMR (376 MHz, DMSOR) δ ppm -113.48 (s, 1 F). 46-1 547.2 Ή NMR (400 MHz, DMSOR) δ ppm 8.42-8.47 (m, 1 H), 8.39 (d, /=5.0 Hz, 1 H), 7.48-7.55 (m , 1 H), 7.21-7.33 (m, 4 H), 6.80-6.91 (m, 1 H), 6.21 (br d, /=17.0 Hz, 1 H) , 5.745.79 (m, 1 H), 4.93 (br s, 1 H), 3.98-4.44 (m, 3 H), 3.533.88 (m, 3 H), 2.33- 2.45 (m, 2 H), 1.97 (s, 3 H), 1.34 (d, /=6.6 Hz, 3 H), 1.03-1.07 (m, 3 H) . ¹⁹ F NMR (376 MHz, DMSO-/ ₆ ) δ ppm -113.84 (s, 1 F). 46-2 547.0 Ή NMR (400 MHz, DMSOR) δ ppm 8.48 (br s, 1 H), 8.39 (d, /=5.0 Hz, 1 H), 7.47-7.56 (m, 1 H ), 7.21-7.33 (m, 4 H), 6.80-6.92 (m, 1 H), 6.21 (br d, /=16.4 Hz, 1 H), 5, 77 (br d, /=11.4 Hz, 1 H), 4.97 (br s, 1 H), 4.00-4.48 (m, 3 H), 3.77 (td, /=12 ,2, 6.1 Hz, 2 H), 3.04-3.33 (m, 1 H), 2.35-2.47 (m, 2 H), 1.97 (s, 3 H), 1.33 (br d, /=6.6 Hz, 3 H), 1.00-1.08 (m, 3 H). ¹⁹ F NMR (376 MHz, DMSO-/ ₆ ) δ ppm -113.86 (s, 1 F).

- 202 044387

47 562.1 Ή NMR (400 MHz, DMSO-U) δ ppm 9.01 (s, 1 H), 8.48 (br d, 7=5.0 Hz, 1 H), 7.50-7.57 (m, 1 H), 7.27-7.35 (m, 3 H), 7.20-7.27 (m, 1 H), 6.79-6.93 (m, 1 H), 6.17- 6.26 (m, 1 H), 5.65-5.80 (m, 1 H), 4.92-5.02 (m, 1 H), 4.23-4.44 (m, 2 H ), 3.99-4.20 (m, 1 H), 3.72-3.88 (m, 1 H), 3.403.71 (m, 1 H), 2.35-2.48 (m, 4 H), 1.36 (d, 7=6.6 Hz, 3 H), 1.06 (t, 7=7.5 Hz, 6 H). ¹⁹ F NMR (377 MHz, DMSO-b/D δ ppm-114.01 (s, 1 F). 48 569.2 Ή NMR (400 MHz, DMSO 7 ₆ ) δ ppm 10.10 (s, 1 H), 9.01 (s, 1 H), 8.37 (br s, 1 H), 7.22-7.32 (m, 1 H), 6.80-6.93 (m, 1 H), 6.66-6.77 (m, 2 H), 6.20 (br d, 7=16.6 Hz, 1 H), 5.715.81 (m, 1 H), 4.83-5.04 (m, 1 H), 3.98-4.45 (m, 3 H), 3.56-3.87 (m , 2 H), 3.07 (br dd, 7=4.1, 2.9 Hz, 1 H), 2.592.70 (m, 1 H), 1.33 (br d, 7=5.8 Hz , 3 H), 1.07-1.14 (m, 3 H), 1.01 (br d, 7 = 6.6 Hz, 3 H). ¹⁹ F NMR (376 MHz, DMSO-U) δ ppm -115.41 (br s, 1 F). 49-1 576.2 Ή NMR (400 MHz, DMSO-J ₆ ) δ ppm 10.05 (s, 1 Η), 8.40 (br s, 1 H), 7.54 (br d, 7=7.7 Hz, 1 H ), 7.17-7.33 (m, 3 H), 6.78-7.01 (m, 2 H), 6.61-6.75 (m, 2 H), 6.20 (br d , 7=15.6 Hz, 1 H), 5.76 (br d, 7=10.8 Hz, 1 H), 4.86 (br s, 1 H), 4.264.52 (m, 1 H) , 3.96-4.25 (m, 2 H), 3.36-3.74 (m, 2 H), 2.93-3.13 (m, 1 H), 1.28 (br s, 3 H), 1.11 (br s, 9 H). 49-2 576.2 Ή NMR (400 MHz, DMSO-U) δ ppm 10.07 (br s, 1 H), 8.37 (br s, 1 H), 7.54 (br d, 7=7.9 Hz, 1 H ), 7.25-7.33 (m, 1 H), 7.17-7.25 (m, 2 H), 6.96 (br d, 7 = 6.0 Hz, 1 H), 6.776, 90 (m, 1 H), 6.71 (d, 7=8.3 Hz, 1 H), 6.65 (br t, 7=8.7 Hz, 1 H), 6.20 (br d, 7=16.6 Hz, 1 H), 5.72-5.80 (m, 1 H), 4.85 (br s, 1 H), 4.18-4.43 (m, 2 H), 3.96-4.17 (m, 1 H), 3.70 (br s, 1 H), 3.41-3.64 (m, 1 H), 3.22 (br s, 1 H), 1.33 (br d, 7=6.2 Hz, 3 H), 1.08-1.14 (m, 9 H). 50-1 546.8 NMR (400 MHz, DMSO-7 ₆ ) δ ppm 7.51-7.61 (m, 1 H), 7.43 (dd, 7 = 7.9, 1.0 Hz, 1 H), 7.33 -7.39 (m, 2 H), 7.287.33 (m, 2 H), 7.24 (td, 7=7.6, 1.4 Hz, 1 H), 7.12 (dd, 7= 7.9, 1.0 Hz, 1 H), 6.90 (br dd, 7=16.3, 10.5 Hz, 1 H), 6.21 (br d, 7=16.6 Hz, 1 H), 5.76 (dd, 7=8.7, 1.4 Hz, 1 H), 5.13-5.59

- 203 044387

50-2 546.8 (m, 1 Η), 4.64-4.94 (m, 1 Η), 3.99-4.41 (m, 3 Η), 3.473.90 (m, 2 Η), 2.75 (quin , 7=6.8 Hz, 1 Η), 1.25 (br s, 3 Η), 1.11 (d, 7=6.8 Hz, 3 Η), 0.99 (d, 7=6, 8 Hz, 3 Η). Ή NMR (400 MHz, DMSO-7,) δ ppm 7.50-7.61 (m, 1 Η), 7.44 (dd, 7=7.7, 0.8 Hz, 1 Η), 7, 33-7.39 (m, 2 Η), 7.287.33 (m, 2 Η), 7.24 (td, 7=7.5, 1.3 Hz, 1 Η), 7.13 (dd, 7 =7.9, 0.8 Hz, 1 Η), 6.89 (br dd, 7=16.3, 10.7 Hz, 1 Η), 6.21 (br d, 7=16.6 Hz, 1 Η), 5.76 (dd, 7=9.9, 1.2 Hz, 1 Η), 5.13-5.59 (m, 1 Η), 4.69-4.92 (m, 1 Η), 3.97-4.39 (m, 3 Η), 3.613.75 (m, 1 Η), 3.44-3.60 (m, 1 Η), 2.64-2.73 (m , 1 Η), 1.25 (br s, 3 H), 1.09 (d, 7=6.8 Hz, 3 H), 1.00 (d, 7=6.8 Hz, 3 H). 51-1 526.8 Ή NMR (400 MHz, DMSO-7 ₆ ) δ ppm 7.48-7.59 (m, 1 H), 7.43 (dd, 7 = 8.3, 1.0 Hz, 1 H), 7, 32-7.39 (m, 2 H), 7.257.32 (m, 2 H), 7.22 (dd, 7=7.5, 1.7 Hz, 1 H), 7.12 (dd, 7 =7.7, 1.0 Hz, 1 H), 6.82-6.94 (m, 1 H), 6.21 (br d, 7 = 16.0 Hz, 1 H), 5.76 ( dd, 7=10.4, 2.1 Hz, 1 H), 4.70-5.56 (m, 1 H), 3.95-4.41 (m, 3 H), 3.45-3 .79 (m, 3 H), 2.55-2.65 (m, 1 H), 2.43 (d, 7=1.0 Hz, 3 H), 1.25 (br s, 3 H) , 1.08 (d, 7=6.8 Hz, 3 H), 0.98 (d, 7=6.8 Hz, 3 H). 51-2 526.8 Ή NMR (400 MHz, DMSO-7 ₆ ) δ ppm 7.49-7.60 (m, 1 H), 7.42 (dd, 7 = 7.7, 1.7 Hz, 1 H), 7, 30-7.38 (m, 2 H), 7.267.30 (m, 2 H), 7.23 (td, 7=7.5, 1.3 Hz, 1 H), 7.10 (dd, 7 =7.8, 0.9 Hz, 1 H), 6.70-6.97 (m, 1 H), 6.21 (br d, 7 = 16.6 Hz, 1 H), 5.76 ( dd, 7=10.5, 2.2 Hz, 1 H), 4.73-5.50 (m, 1 H), 3.89-4.79 (m, 3 H), 3.31-3 .73 (m, 3 H), 2.63-2.73 (m, 1 H), 2.43 (d, 7=1.0 Hz, 3 H), 1.25 (br s, 3 H) , 1.10 (d, 7=6.8 Hz, 3 H), 0.97 (d, 7=6.8 Hz, 3 H). 52-1 541.8 Ή NMR (400 MHz, DMSO-7 ₆ ) δ ppm 9.94 (br s, 1 H), 8.12 (br s, 1 H), 7.38 (dd, 7=7.5, 1.7 Hz, 1 H), 7.31 (td, 7=7.7, 1.5 Hz, 1 H), 7.15-7.24 (m, 2 H), 7.06 (dd, 7=6 ,0, 1.0 Hz, 1 H), 6.78-6.95 (m, 1 H), 6.70 (d, 7=8.3 Hz, 1 H), 6.64 (t, 7 =8.8 Hz, 1 H), 6.21 (dd, 7 = 16.7, 5.3 Hz, 1 H), 5.77 (dd, 7 = 10.4, 2.3 Hz, 1 H ), 4.80 (br s, 1 H), 4.11-4.48 (m, 4 H), 3.45-3.67 (m, 2 H), 3.07-3.25 (m , 1 H), 2.12 (s, 3 H), 1.36 (d, 7=6.6 Hz, 3 H), 1.05 (d, 7=6.6 Hz, 3 H), 0 .95 (d, 7=6.8

- 204 044387

Hz, 3 N). 52-2 541.8 Ή NMR (400 MHz, DMSO-7 ₆ ) δ ppm 9.94 (s, 1 Η), 8.15 (s, 1 Η), 7.38 (dd, 7=8.5, 1.0 Hz, 1 Η), 7.32 (td, 7=6.6, 1.7 Hz, 1 Η), 7.16-7.25 (m, 2 Η), 7.06 (dd, 7=6.8 , 1.2 Hz, 1 Η), 6.81-6.93 (m, 1 Η), 6.70 (d, 7=8.3 Hz, 1 Η), 6.63 (t, 7=8 .8 Hz, 1 Η), 6.20 (br dd, 7=15.8, 5.4 Hz, 1 Η), 5.77 (dd, 7=10.2, 2.5 Hz, 1 Η) , 4.87-4.98 (m, 1 Η), 3.98-4.47 (m, 3 Η), 3.57-3.80 (m, ZN), 2.97-3.11 ( m, 1 Η), 2.12 (s, 3 Η), 1.30 (br d, 7=6.4 Hz, 3 Η), 1.06 (d, 7=6.8 Hz, 3 Η) , 0.96 (d, 7=6.6 Hz, ZN). 53-1 550.2 Ή NMR (400 MHz, DMSOA) δ ppm 8.45 (br s, 1 Η), 7.49-7.60 (m, 1 Η), 7.22-7.41 (m, 4 Η), 6 .78-6.93 (m, 1 Η), 6.21 (br d, 7=16.2 Hz, 1 Η), 5.72-5.81 (m, 1 Η), 4.98 (br s, 1 H), 4.22-4.46 (m, 2 H), 3.99-4.21 (m, 2 H), 3.373.90 (m, 2 H), 2.99-3, 28 (m, 1 H), 1.71 (s, 3 H), 1.33 (br d, 7=6.6 Hz, 3 H), 1.21 (br dd, 7=33.2, 6 .4 Hz, 6 H). ¹⁹ F NMR (376 MHz, DMSO 7 ₆ ) δ ppm -113.97 (s, 1 F). 53-2 550.2 NMR (400 MHz, DMSO-7 ₆ ) δ ppm 8.43 (br s, 1 H), 7.55 (q, 7 = 6.2 Hz, 1 H), 7.24-7.40 (m, 4 H), 6.79-6.93 (m, 1 H), 6.21 (br d, 7=16.2 Hz, 1 H), 5.77 (br d, 7=11.8 Hz, 1 H), 4.95 (br s, 1 H), 4.23-4.47 (m, 2 H), 3.98-4.21 (m, 2 H), 3.39-3.89 (m, 2 H), 3.04-3.28 (m, 1 H), 1.71 (s, 3 H), 1.35 (br d, 7=6.6 Hz, 3 H), 1 .21 (br dd, 7=35.5, 6.4 Hz, 6 H). ¹⁹ F NMR (376 MHz, DMSOA) δ ppm -113.94 (s, 1 F).

Biological analysis data.

For connections in table. 15 used the following analysis conditions.

Analysis of exchange of bound nucleotides. Purified GDP-bound KRAS protein (amino acids 1-169), containing both G12C and C118A amino acid substitutions and an N-terminal His tag, was preincubated with a dose-matched compound for 2 hours in assay buffer (25 mM HEPES pH 7.4, 10 mM MgCl ₂ and 0.01% Triton X-100). After preincubation with the compound, purified SOS protein (amino acids 564-1049) and GTP (Roche 10106399001) were added to the assay wells and incubated for an additional hour. To determine the degree of inhibiting SOS-mediated nucleotide metabolism to the analytical holes, the purified GST-Laughter CRAF (amino acids 1-149), acceptor granules with nickel helat Alphalisa (Perkinelmer AL108R) and donor granules with glutation are added Elmer 6765302) and incubated within 10 min. The assay plates were then read on a PerkinElmer EnVision Multilabel Reader using AlphaScreen® technology and the data were analyzed using a 4-parameter logistic model to calculate IC ₅₀ values.

Phospho-ERK1/2 assay using MSD.

MIA PaCa-2 (ATCC® CRL-1420™) and A549 (ATCC® CCL-185™) cells were cultured in RPMI 1640 medium (ThermoFisher Scientific 11875093) containing 10% fetal bovine serum (ThermoFisher Scientific 16000044) and 1x penicillin-strepto mycin -glutamine (ThermoFisher Scientific 10378016). 16 hours before compound treatment, MIA PaCa-2 or A549 cells were seeded in 96-well cell culture plates at a density of 25,000 cells/well and incubated at 37°C, 5% CO2. The compound was dose-dependently diluted in growth medium, added to the appropriate wells of the cell culture plate and then incubated at 37°C, 5% CO ₂ for 4 hours. After treatment with the compound, cells were stimulated with 10 ng/ml EGF (Roche 11376454001) for 10 min, washed with ice-cold Dulbecco's phosphate buffered saline solution containing no Ca ²⁺ or Mg ²⁺ (ThermoFisher Scientific 14190144), and then lysed in RIPA buffer (50 mM Tris-HCl pH 7.5 , 1% Igepal, 0.5% sodium dodecyl sulfate, 150 mM NaCl and 0.5% sodium deoxycholate) containing protease inhibitors (Roche 4693132001) and phosphatase inhibitors (Roche 4906837001). Cell lysates were stored frozen at -80°C overnight. Phosphorylation of ERK1/2 in compound-treated lysates was analyzed using kits

- 205 044387

Phospho-ERK1/2 Whole Cell Lysate (Meso Scale Discovery K151DWD) according to the manufacturer's protocol. Assay plates were read on a Meso Scale Discovery Sector Imager 6000 and data were analyzed using a 4-parameter logistic model to calculate IC ₅₀ values.

Table 15

Biochemical and cellular activity of compounds

No. note 1C ₅₀ (µM) for coupled exchange 1C ₅₀ for p-ERK (MIA PaCa-2, µM) 1C ₅₀ for p-ERK (A549, µM) 1-1 0.355 2.55 >33.3 1-2 2.38 6.08 >100 1-3 0.610 3.84 >100 1-4 >10 >100 >100 1-5 6.66 - - 1-6 2.88 36.2 >100 1-7 0.209 1.86 >100 1-8 0.894 6.07 >100 1-9 5.92 - - 1-10 0.381 1.09 11.1 1-11 0.695 8.2 >100 1-12 I,8 - - 1-13 >10 - - 1-14 1.78 5.03 >100 1-15 0.562 4.7 >100 1-16 0.492 4.83 >100 1-17 63.5 - - 1-18 0.370 0.559 >33.3 1-19 0.297 1.33 >100 1-19-1 0.115 0.368 >100 1-19-2 5.10 >100 >100 1-20 0.683 4.99 >100 1-21 1.30 4.89 >100 1-22 >250 - - 1-23 >250 - - 1-28 2.20 - >100 2-1 0.341 1.89 3.7 2-2 12.7 - - 2-3 4.05 6.53 >100 2-4 >250 - — 2-5 0.684 5.46 3.7 2-5-1 0.308 1.14 >100 2-5-2 1.35 7.48 3.7 2-6 1.59 2.97 3.7 2-6-1 13.0 - - 2-6-2 1.25 1.29 3.7 2-7 1.08 3.87 >33.3 2-8 0.361 0.258 >100 2-9 0.301 0.747 >100

- 206 044387

2-10 1.73 3.07 >100 3-1 0.266 3.23 >100 3-1-1 3.00 >100 >100 3-1-2 0.302 2.35 >100 3-2 >250 - - 3-3 11.3 - - 3-4 >250 - - 3-5 0.693 5.26 3.7 3-6 1.05 11.8 >100 3-7 6.98 - 3-8 1.07 7.05 >100 3-9 3.37 9.25 >33.3 3-10 4.74 66.4 11.1 3-11 0.457 3.06 11.1 3-12 2.56 7.66 >100 3-13 6.49 - - 3-14 5.64 - - 3-15 4.03 20.1 >100 3-16 2.60 21.3 >100 3-17 5.48 - - 3-18 2.60 >100 >100 3-19 0.954 2.03 >33.3 3-20 2.99 9.65 >100 3-21 32.0 - - 3-22 0.249 1.12 >33.3 3-23 4.65 13.6 >100 3-24 9.07 23.7 >100 3-25 >250 - - 4-1 0.529 2.34 >100 4-2 >250 - - 4-3 >250 - - 4-4 >250 - -

-207 044387

4-5 >250 - - 4-6 0.630 10.3 >100 4-7 125 - - 4-8 177 - - 4-9 >250 - - 5-1 0.875 2.86 >100 5-2 14.2 - - 5-3 14.2 - - 5-4 0.610 3.25 >100 5-5 0.341 2.53 >100 5-6 0.883 5.9 >100 5-7 0.815 3.79 >100 5-8 0.433 1.2 >33.3 5-9 0.139 0.822 >100 6-1 0.537 1.3 3.7 6-2 5.31 - - 7-1 0.299 0.43 >100 7-2 0.180 0.222 >100 7-3 1.73 5.83 >100 8-1 0.542 0.211 62.3 8-1-1 0.172 0.046 69.8 8-1-2 0.322 0.811 >100 8-2 0.152 0.050 >100 8-3 0.283 0.061 >100 8-3-1 0.282 0.408 >100 8-3-2 0.340 0.028 >100 8-4 0.095 0.017 >33.3 8-5 0.400 2.41 >100 8-6 0.100 0.012 69.9 8-6-2 0.185 0.128 >100 8-6-1 0.066 0.01 >33.3 9-1 0.155 0.052 57.4

- 208 044387

9-2 0.289 1.11 74.4 9-3 0.113 0.035 66.1 9-4 0.198 0.023 13 9-5 1.33 3.92 >100 9-6 0.237 3.51 >100 9-7-2 - 0.232 62.0 9-7-1 - 0.023 14.9 9-9 0.147 0.136 65.6 9-10 0.101 0.117 66.5 9-11 0.093 0.147 64 9-12 1.32 1.29 >100 9-13 0.306 0.078 12.8 9-14 0.129 0.344 >100 10-1 24.5 - - 10-2 1.93 36.1 >100 10-3 ι,ιο 9.33 >100 10-4 0.235 3.47 >100 10-5 0.297 3.41 >100 10-6 1.20 6.07 >100 10-7 0.533 9.2 >100 10-8 1.34 12.9 >100 10-9 1.68 33.6 >100 10-10 0.359 6.85 >100 10-11 0.516 14.7 >100 10-12 0.912 22.6 >100 10-13 15.1 - - 11-1-1 0.231 0.247 42.9 11-1-2 0.151 0.016 21.7 11-2-1 0.219 0.054 10 11-2-2 0.256 0.006 26.4 12 3.34 25.9 >100 13 5.20 >100 >100

- 209 044387

14 2.69 - - 15 2.56 - - 16 2.93 8.62 33.3 17-1 20.6 - - 17-2 1.02 2.6 49.1 18-1 23.0 - - 18-2 0.760 >100 >100 18-3 24.6 - - 19-1 15.2 - - 19-2 1.62 4.69 3.7 19-3 115 - - 20 78.8 - - 21 9.41 - - 22 0.927 56.1 >100 23 3.29 3.21 >33.3 24 6.20 - - 25 0.251 0.786 >100 26 3.90 30.3 >100 27 1.28 12.8 >33.3 28 8.17 - - 29 3.76 >100 >100 thirty 7.51 58 >100 31 123 - - 32 28.6 - - 33 103 - - 34 34.2 - - 35 8.01 79.7 >100 36 39.8 - - 37 9.07 14.7 >100 38 0.484 1.39 >33.3

For connections in table. 16 used the following analysis conditions. Analysis of exchange of bound nucleotides. Purified GDP-bound KRAS protein (amino acids 1–169), containing both G12C and C118A amino acid substitutions and an N-terminal His tag, was preincubated with a dose-matched compound for 5 min in assay buffer (25 mM HEPES pH 7.4, 10 mM MgCl ₂ and 0.01% Triton X-100). After preincubation with the compound, purified SOS protein (amino acids 564-1049) and GTP (Roche 10106399001) were added to the assay wells and incubated for an additional 30 min. To determine the degree of inhibition of SOS-mediated nucleotide exchange, purified GST-tagged cRAF (amino acids 1-149), AlphaLISA nickel chelate acceptor beads (PerkinElmer AL108R), and AlphaScreen glutathione donor beads (PerkinElmer 6765302) were added to assay wells and incubated for 5 minutes. The assay plates were then read on a PerkinElmer EnVision Multilabel Reader using AlphaScreen® technology and the data were analyzed using a 4-parameter logistic model to calculate IC ₅₀ values.

Phospho-ERK1/2 assay using MSD. MIA PaCa-2 (ATCC® CRL-1420™) and A549 (ATCC® CCL-185™) cells were cultured in RPMI 1640 medium (ThermoFisher Scientific 11875093) containing 10% fetal bovine serum (ThermoFisher Scientific 16000044) and 1x penicillin-strepto mycin -glutamine (ThermoFisher Scientific 10378016). Sixteen hours before compound treatment, MIA PaCa-2 or A549 cells were seeded in 96-well cell culture plates at a density of 25,000 cells/well and incubated at 37°C, 5% CO ₂ . When selecting a dose-dependent effect, the compound was diluted in growth medium, added to the appropriate wells of the cell culture plate, and then incubated at 37°C, 5% CO ₂ for 2 hours. After treatment with the compound, the cells were stimulated with 10 ng/ml EGF ( Roche 11376454001) for 10 min, washed with ice-cold Dulbecco's phosphate buffered saline solution containing no Ca ²⁺ or Mg ²⁺ (ThermoFisher Scientific 14190144), and then lysed in RIPA buffer (50 mM Tris-HCl pH 7, 5, 1% Igepal, 0.5% sodium dodecyl sulfate, 150 mM NaCl and 0.5% sodium deoxycholate) containing protease inhibitors (Roche 4693132001) and phosphatase inhibitors (Roche 4906837001). Phosphorylation of ERK1/2 in compound-treated lysates was analyzed using Phospho-ERKl/2 Whole Cell Lysate kits (Meso Scale Discovery K151DWD) according to the manufacturer's protocol. Assay plates were read on a Meso Scale Discovery Sector Imager 6000 and data were analyzed using a 4-parameter logistic model to calculate _1C50 values.

Table 16

Biochemical and cellular activity of compounds

No. note 1C ₅₀ (µM) for coupled exchange 1C ₅₀ for p-ERK (MIA PaCa-2, µM) 1C ₅₀ for p-ERK (A549, µM) 39 0.063 0.029 >100 39-1 0.025 0.028 - 39-2 0.222 0.195 - 40 0.063 0.029 - 40-1 0.057 0.022 - 40-2 0.597 0.321 - 41 0.136 0.085 - 41-1 0.093 0.072 - 41-2 1.39 1.66 - 42 0.335 0.362 - 42-1 0.150 0.103 - 42-2 1.56 - - 43 0.038 0.034 - 44 0.103 0.065 - 44-1 0.033 0.025 - 44-2 0.208 0.138 - 45 0.319 0.062 - 45-1 0.089 0.049 - 45-2 0.727 0.435 - 46 0.493 0.169 - 46-1 0.170 0.126 - 46-2 0.971 0.552 - 47 0.409 0.152 - 48 0.081 0.063 >10 49 0.177 0.154 - 49-1 2.58 - - 49-2 0.117 0.051 - 50 0.276 0.188 - 50-1 0.117 0.097 - 50-2 3.69 - - 51 0.538 0.440 - 51-1 AND 2 - - 51-2 0.221 0.089 - 52 0.338 0.215 - 52-1 5.99 - - 52-2 0.146 0.177 - 53 0.125 0.071 - 53-1 0.098 0.093 - 53-2 0.320 0.229 -

(-) = not researched.

The present invention has been described using preferred embodiments. However, it should be understood that the present invention is not limited to the disclosed embodiments. It is understood that, given the description of the embodiments of the present invention given herein, various modifications can be made by one skilled in the art. Such modifications are covered by the claims below.

Claims (24)

1. Connection, where the specified connection is selected from: or a pharmaceutically acceptable salt thereof. 2. A pharmaceutically acceptable salt of a compound, wherein said compound is 3. A connection, where said connection is 4. A pharmaceutically acceptable salt of atropisomer 1 of the compound according to claim 2, corresponding to the 1st elution isomer, which can be obtained by separating atropisomers using supercritical liquid chromatography using Chiralpak IC, 30x250 mm, 5 μm, 55% MeOH/CO ₂ , 120 g/min, 10.3 MPa (103 bar). 5. A pharmaceutically acceptable salt of atropisomer 2 of the compound according to claim 2, corresponding to the 2nd elution isomer, which can be obtained by separating atropisomers using supercritical liquid chromatography using Chiralpak IC, 30x250 mm, 5 μm, 55% MeOH/CO2, 120 g/min, 10.3 MPa (103 bar). 6. Atropisomer 1 of the compound according to claim 3, corresponding to the 1st elution isomer, which can be obtained by separating atropisomers using supercritical liquid chromatography using Chiralpak IC, 30x250 mm, 5 μm, 55% MeOH/CO ₂ , 120 g/ min, 10.3 MPa (103 bar). 7. Atropisomer 2 of the compound according to claim 3, corresponding to the 2nd elution isomer, which can be obtained by separating atropisomers using supercritical liquid chromatography using Chiralpak IC, 30x250 mm, 5 μm, 55% MeOH/CO ₂ , 120 g/ min, 10.3 MPa (103 bar). - 212 044387 8. A pharmaceutically acceptable salt according to claim 2, wherein the compound is 9. A pharmaceutically acceptable salt according to claim 2, wherein the compound is 10. The connection according to claim 3, where the connection is 11. The connection according to claim 3, where the connection is 12. A pharmaceutical composition for the treatment of cancer, comprising a compound according to any one of claims 1 to 11 and a pharmaceutically acceptable excipient. 13. An in vitro method of inhibiting KRAS G12C in a cell, comprising bringing the cell into contact with a compound according to any one of claims 1 to 11 or a composition according to claim 12. 14. A method of treating cancer in a subject in need of the treatment, comprising administering to the subject a therapeutically effective amount of a compound of any one of claims 1 to 11 or a composition of claim 12. 15. The method of claim 14, wherein the cancer is non-small cell lung cancer, colorectal cancer, pancreatic cancer, appendix cancer, endometrial cancer, stomach cancer, small bowel cancer, nasal cancer, bile duct cancer or skin cancer. 16. The method of claim 14, wherein the cancer is non-small cell lung cancer. 17. The method of claim 14, wherein the cancer is pancreatic cancer. 18. The method of claim 14, wherein the cancer is colorectal cancer. 19. The method according to any one of claims 14 to 18, wherein the cancer is mediated by the G12C mutation of KRAS. 20. A connection, where said connection is 21. A connection, wherein said connection is 22. A connection, wherein said connection is - 213 044387 23. A connection, where said connection is 24. A connection, wherein said connection is