EA046336B1 - 3,6-DIAMINOPYRIDAZIN-3-YL DERIVATIVES, CONTAINING THEIR PHARMACEUTICAL COMPOSITIONS AND THEIR APPLICATION AS PRO-APOPTOTIC AGENTS - Google Patents

Description

Field of technology to which the invention relates

The present invention relates to 3,6-diaminopyridazin-3-yl derivatives, pharmaceutical compositions containing them and their use as proapoptotic agents. The compounds of the present invention inhibit the activity of the Bcl-xL protein and may be of interest for the treatment of malignancies, immunopathological and autoimmune diseases.

Prerequisites for creating the invention

Apoptosis (programmed cell death) is an evolutionarily conserved pathway critical for tissue homeostasis, development, and removal of damaged cells. Deregulation of apoptosis contributes to the development of human diseases, including malignancies, neurodegenerative disorders, immune system diseases, and autoimmune diseases (Hanahan and Weinberg, Cell. 2011 Mar 4; 144(5):646-74; Marsden and Strasser, Annu Rev Immunol. 2003; 21:71105; Vaux and Flavell, Curr Opin Immunol. 2000 Dec; 12(6):719-24). Evasion of apoptosis is regarded as a characteristic feature of malignancy, participating in the development, as well as in the sustained growth of tumors, and in the emergence of resistance to anticancer therapies (Hanahan and Weinberg, Cell. 2000 Jan 7; 100(1):57-70) .

The Bcl-2 family of proteins includes key regulators of cell viability that can suppress (eg, Bcl-2, Bcl-xL, Mcl-1) or stimulate (eg, Bad, Bax) apoptosis (Gross et al., Genes Dev. 1999 Aug. 1;13(15):1899-911, Youle and Strasser, Nat. Rev. Mol. Cell Biol. 2008 Jan;9(1):47-59).

In the light of stress stimuli, whether a cell survives or undergoes apoptosis depends on the degree of pairing between Bcl-2 family members that stimulate cell death and family members that stimulate cell survival. In most cases, these interactions involve docking of the Bcl-2 homology 3 (BH3) domain of pro-apoptotic family members into a groove on the surface of pro-survival members. The presence of a Bcl-2 homology (BH) domain determines membership in the Bcl-2 family, which is divided into three main groups depending on which specific BH domains are present in the protein. Pro-survival members such as Bcl2, Bcl-xL, and Mcl-1 contain BL domains 1–4, whereas Bax and Bak, proapoptotic effectors of outer mitochondrial membrane permeabilization during apoptosis, contain BL domains 1–3 (Youle and Strasser, Nat Rev Mol Cell Biol 2008 Jan;9(1):47-59).

Overexpression of pro-survival members of the Bcl-2 family is a hallmark of malignancy, and these proteins have been shown to play important roles in tumor development, maintenance, and resistance to antitumor therapy (Czabotar et al., Nat. Rev. Mol. Cell Biol. 2014 Jan;15(1):49-63). Bcl-xL (also called BCL2L1, from BCL2-like 1 (BCL2like 1)) is often amplified in cancer (Beroukhim et al., Nature 2010 Feb 18; 463(7283):899-905), and has been shown to expression inversely correlates with sensitivity to more than 120 antitumor therapeutic molecules in a representative panel of cancer cell lines (NCI-60) (Amundson et al., Cancer Res. 2000 Nov 1; 60(21):6101-10).

In addition, several studies using transgenic gene knockout mouse models and transgenic overexpression of Bcl-2 family members have highlighted the importance of these proteins in immune system diseases and autoimmune diseases (for review, see Merino et al., Apoptosis 2009 Apr; 14(4):570-83. doi: 10.1007/s10495-008-0308-4. PMID: 19172396). Transgenic overexpression of Bcl-xL in the T cell compartment resulted in resistance to apoptosis induced by glucocorticoids, gamma irradiation, and CD3 cross-linking, suggesting that overexpression of transgenic Bcl-xL may reduce apoptosis in resting and activated T cells (Droin et al., Biochim Biophys Acta 2004 Mar l;1644(2-3):179-88. doi: 10.1016/j.bbamcr.2003.10.011.PMID: 14996502). Consistent or high expression of antiapoptotic Bcl-2 family proteins was observed in patient samples (Pope et al., Nat Rev Immunol. 2002 Jul; 2(7):527-35. doi: 10.1038/nri846.PMID: 12094227) . In particular, T cells isolated from the joints of patients with rheumatoid arthritis showed increased expression of Bcl-xL and were resistant to spontaneous apoptosis (Salmon et al., J Clin Invest. 1997 Feb 1; 99(3):439-46. doi: 10.1172/JCI119178.PMID: 9022077). The use of BH3 mimetics has shown benefit in preclinical models of immune system diseases and autoimmune diseases. Treatment with ABT-737 (an inhibitor of Bcl-2, Bcl-xL and Bcl-w) resulted in a strong inhibition of lymphocyte proliferation in vitro. It is important to note that mice treated with ABT-737 in studies in animal models of arthritis and lupus showed a significant reduction in disease severity (Bardwell et al., J Clin Invest. 1997 Feb 1; 99(3):439-46. doi: 10.1172 /JCI119178.PMID: 9022077). In addition, ABT-737 was shown to prevent allogeneic T cell activation, proliferation, and cytotoxicity in vitro and suppress allogeneic T and B cell responses after skin transplantation with high selectivity for lymphoid cells (Cippa et al., Transpl Int 2011 Jul;24(7):722-32. doi: 10.1111/j.1432-2277.2011.01272.x. Epub 2011 May 25. PMID: 21615547).

The above research results served as an impetus for the discovery and development of a new class of drugs called BH3 mimetics. These molecules are capable of disrupting the interaction between proapoptotic and antiapoptotic members of the Bcl-2 family and are

- 1 046336 powerful inducers of apoptosis. This new class of drugs includes inhibitors of Bcl-2, Bcl-xL, Bcl-w and Mcl-1. The first described BH3 mimetics were ABT-737 and ABT-263, targeting Bcl-2, Bcl-xL and Bcl-w (Park et al. J. Med. Chem. 2008 Nov 13; 51(21):6902-15 ; Roberts et al., J. Clin. Oncol. 2012 Feb 10; 30(5):488-96). Subsequently, selective Bcl-2 inhibitors were also discovered (ABT-199 and S55746 - Souers et al., Nat Med. 2013 Feb; 19(2):202-8; Casara et al., Oncotarget 2018 Apr 13; 9(28 ):20075-20088), Bcl-xL (A-1155463 and A-1331852 - Tao et al., ACS Med Chem Lett. 2014 Aug 26; 5(10):1088-93; Leverson et al., Sci Transl Med 2015 Mar 18;7(279):279ra40) and Mcl-1 (A-1210477, S63845, S64315, AMG-176 and AZD-5991 - Leverson et al., Cell Death Dis. 2015 Jan 15;6:e1590. ; Kotschy et al., Nature 2016, 538, 477-482; Maragno et al., AACR 2019, Poster #4482; Kotschy et al., WO 2015/097123; Caenepeel et al., Cancer Discov. 2018 Dec; 8( 12):1582-1597; Tron et al., Nat. Commun. 2018 Dec 17; 9(1):5341). The selective Bcl-2 inhibitor ABT-199 is currently approved for the treatment of patients with CLL and AML as a combination therapy, while other inhibitors are still in preclinical or clinical development. In preclinical models, ABT-263 has demonstrated activity in several hematologic malignancies and solid tumors (Shoemaker et al., Clin. Cancer Res. 2008 Jun 1; 14(11):3268-77; Ackler et al., Cancer Chemother. Pharmacol 2010 Oct;66(5):869-80; Chen et al., Mol. Cancer Ther. 2011 Dec;10(12):2340-9). In clinical studies, ABT-263 demonstrated objective antitumor activity in lymphoid malignancies (Wilson et al., Lancet Oncol. 2010 Dec; 11(12):1149-59; Roberts et al., J. Clin. Oncol. 2012 Feb 10 ; 30(5):488-96), and its activity is being studied in combination with several methods of treating solid tumors. Selective Bcl-xL inhibitors, A-1155463 or A-1331852, have demonstrated in vivo activity in preclinical models of T-ALL (T-cell acute lymphoblastic leukemia) and various types of solid tumors (Tao et al., ACS Med. Chem. Lett. 2014 Aug 26; 5(10):1088-93; Leverson et al., Sci. Transl. Med. 2015 Mar 18; 7(279):279ra40). Selective Mcl-1 inhibitors have shown promising in vivo activity in several types of hematologic malignancies in preclinical models, and three of them, inhibitors S64315, AMG176, and AZD5991, are currently being investigated in clinical trials (Yang et al., Eur. J. Med Chem 2019 May 8;177:63-75). Thus, BH3 mimetics represent an extremely attractive approach for the development of new treatments in oncology, as well as in the field of immunopathological and autoimmune diseases. In particular, there is a need for small molecules that selectively inhibit the Bcl-xL protein. The present invention satisfies this need.

Summary of the Present Invention

The present invention provides potent selective Bcl-xL inhibitors of formula (I) as defined below. The present application shows that these compounds are capable of inducing apoptosis of cancer cells in vivo, triggering tumor regression in mice. Due to their pro-apoptotic properties, the compounds of the present invention may be of interest for the treatment of pathologies involving deregulation of apoptosis, such as, for example, malignancies, autoimmune diseases and diseases of the immune system. In addition, these compounds were well tolerated in mice without clinically significant weight loss when treated at effective doses, indicating an acceptable therapeutic window for the use of these small molecules targeting Bcl-xL in cancer treatment. Consistent with the previously described role of Bcl-xL in regulating platelet lifespan (Zhang et al., Cell Death Differ. 2007 Mau; 14(5):943-51; Mason et al., Cell. 2007 Mar 23; 128( 6):1173-86), we observed a decrease in the number of circulating platelets after treatment of mice with these inhibitors and its recovery after cessation of treatment. In view of this effect on platelet survival, the Bcl-xL inhibitors of the present invention can also be used to treat diseases or conditions characterized by excessive or deregulated platelet activity, such as, for example, prothrombotic conditions.

Detailed Description of the Invention

In the first embodiment (E1), the present invention provides compounds of formula (I):

where R1 and R2 independently of each other represent a group selected from hydrogen; linear or branched C1-C ₆ -alkyl, optionally substituted with a hydroxyl or C1-C ₆ -alkoxy group; C3-C ₆ -cycloalkyl; trifluoromethyl; linear or branched C1-C6 alkylene heterocycloalkyl, wherein the heterocycloalkyl group is optionally substituted with a linear or branched C1-C ₆ alkyl group;

or R1 and R2 together with the carbon atoms they carry form a C3-C ₆ -cycloalkylene group,

R ₃ represents a group selected from hydrogen; C ₃ -C ₆ -cycloalkyl; linear or branch

- 2 046336 lennogo C1-C ₆ -alkyl; -X1-NR _a Rb; -X ₁ -N ⁺ R _a RbR _c ; -X1-OR _c ; -X1-COOR _c ; -X ₁ -PO(OH) ₂ ; -X1-SO ₂ (OH); -X1-N3

-Xi-=CH and ,

R _a and Rb independently of each other represent a group selected from hydrogen; heterocycloalkyl; -SO ₂ -phenyl, where phenyl may be replaced by linear or branched ^-^-alkyl; linear or branched ^-^-alkyl, optionally substituted with one or two hydroxyl groups; C ₁ -C ₆ -alkylene-SO ₂ OH; C ₁ -C ₆ -alkylene-SO ₂ O ^- ; C1-C6-alkylene-COOH; C^^-alkylenePO(OH)2; C1-C6-alkylene-NRdRe; C1-C6-alkylene-N ⁺ RdR _e R _f ; ^-^-alkylenephenyl, where phenyl may be replaced by a ^-^-alkoxy group;

groups:

or R _a and Rb together with the nitrogen atom bearing them form the B1 ring; or

R _a , Rb and Rc, together with the nitrogen atom carrying them, form a bridging C ₃ -C ₈ -heterocycloalkyl,

Rc, Rd, Re, R _f independently of each other represent hydrogen or a linear or branched ^-^-alkyl group, or R _d and Re together with the nitrogen atom bearing them form a B2 ring, or

_Rd , Re and Rf, together with the nitrogen atom carrying them, form a bridging C ₃ -C ₈ -heterocycloalkyl,

Het1 is a group selected from:

Het ₂ is a group selected from:

A1 means -NH-, -N(C ₁ -C ₃ -alkyl), O, S or Se,

A ₂ means N, CH or C(R5),

G is selected from the group consisting of:

-C(O)ORg3, -C(O)NRgiRg2, -C(O)Rg2, -NRgiC(O)Rg2, -NRgiC(O)NRgiRg2, -OC(O)NRgiRg2, -NRgiC(O)ORg3, - C(=NORgi)NRgiRg2, -NRgiC(=NCN)NRgiRg2, -NRgiS(O)2NRgiRg2, -S(O) ₂ Rg3,

-S(O)2NRgiRg2, -NRgiS(O)2Rg2, -NRgiC(=NRg2)NRgiRg2, -C(=S)NRgiRg2, -C(=NRgi)NRgiRg2, halogen, -NO2 and -CN, where R _G1 and R _G2 in each case is each independently selected from the group consisting of hydrogen, ^-^-alkyl, optionally substituted with 1-3 halogen atoms, ^-^-alkenyl, ^-^-alkynyl, C ₃ -C ₆ -cycloalkyl, phenyl and -(CH ₂ ) _1-4 -phenyl;

RG3 is selected from the group consisting of ^-^-alkyl, optionally substituted with 1-3 ha atoms

- 3 046336 logen, ^-^-alkenyl, ^-^-alkynyl, C ₃ -C ₆ -cycloalkyl, phenyl and -(CH ₂ )1. ₄ -phenyl; or

RG1 and RG2, together with the atom to which each is attached, combine to form a C ₃ -C ₈ heterocycloalkyl; or, alternatively, G is selected from the group consisting of:

where RG4 is selected from

C1-C ₆ -alkyl, optionally substituted with 1-3 halogen atoms, C2-C6 alkenyl, ^-^-alkynyl and C ₃ -C ₆ -cycloalkyl,

R ₄ represents hydrogen, a fluorine, chlorine or bromine atom, a methyl, hydroxyl or methoxy group,

R5 represents a group selected from ^-^-alkyl, optionally substituted with 1-3 halogen atoms; ^-^-alkenyl; ^-^-alkynyl; halogen or -CN,

R ₆ is a group selected from:

hydrogen;

- ^-^-alkenyl;

- X2-O-R7;

- X2-NSO2-R7;

-C=C(R9)-Yi-O-R7;

C ₃ -C ₆ -cycloalkyl;

C ₃ -C ₆ -heterocycloalkyl, optionally substituted with a hydroxyl group;

C ₃ -C ₆ -cycloalkylene-Y ₂ -R ₇ ;

a C ₃ -C ₆ -heterocycloalkylene-Y ₂ -R ₇ group, a heteroarylene-I ₇ group, optionally substituted by a linear or branched C ₁ -C ₆ -alkyl group,

R ₇ represents a group selected from: a linear or branched C ₁ -C ₆ alkyl group; (C ₃ -C ₆ )cycloalkylene-R ₈ ; or:

- 4 046336

where Su represents C ₃ -C8-cycloalkyl,

Rg represents a group selected from hydrogen; linear or branched C ₁ -C ₆ alkyl, -NR'aR'b;-NR' _a -CO-OR'_c;-NR' _a -CO-R'_c;-N+R'aR'bR'c;-O-R'_c;-NH-X'2-N+R'aR'bR'c;-O-X'2-NR' _a R' _b , -X'2-NR'aR'b, -NR'c-X'2-N3 and:

-NR'c—X'm^CH

R9 represents a group selected from linear or branched C ₁ -C ₆ -alkyl, trifluoromethyl, hydroxyl, halogen, C 1 -C ₆ -alkoxy,

R ₁₀ represents a group selected from hydrogen, fluorine, chlorine, bromine, -CF3 and methyl,

R11 represents a group selected from hydrogen, C ₁ -C ₃ -alkylene-R ₈ , -O-C1-C ₃ -alkylene-K ₈ , -CO-NRhRi and -CH=CH-C1-C4-αalkylene-NRhRi , -CH=CH-CHO, C3-C8-cycloalkylene-CH2-R8, C3-C8heterocycloalkylene- _CH2 ^ ₈ ,

R12 and _R13 independently represent a hydrogen atom or a methyl group,

R14 and R15 independently represent hydrogen or a methyl group, or R14 and R15 together with the carbon atom they carry form cyclohexyl,

R _h and R _i independently of each other represent hydrogen or a linear or branched C1-C ₆ alkyl group,

X1 and _X2 independently represent a linear or branched C1-C6alkylene group, optionally substituted with one or two groups selected from trifluoromethyl, hydroxyl, halogen, C1- _C6 alkoxy,

X' ₂ is a linear or branched C1-C ₆ -alkylene,

R'a and R'b independently of each other represent a group selected from hydrogen; heterocycloalkyl; TO ₂ -phenyl, where phenyl can be replaced by linear or branched C1-C ₆ -alkyl; linear or branched C1-C ₆ -alkyl, optionally substituted with one or two hydroxyl or C1-C ₆ -alkoxy groups; C ₁ -C ₆ -alkylene-SO ₂ OH; C ₁ -C ₆ -αalkylene-SO ₂ O ^- ; C1-C ₆ -alkylene-COOH; C1-C ₆ -alkylene-PO(OH)2; C ₁ -C ₆ -alkylene-NR' _d R'_e;Cr^-alkylene-M^'^'D^;SDS,-al1<ylene-C)-C _g C,alkylene-OH; C1-C ₆ -alkylenephenyl, where phenyl may be replaced by a hydroxyl or C1-C ₆ -alkoxy group;

groups:

or R' _a and R' _b , together with the nitrogen atom bearing them form the B3 ring, or

R' _a , R' _b , and R' _c , together with the nitrogen atom they carry, form a bridging C ₃ -C ₈ -heterocycloalkyl,

R'c, R'd, R'e, R'f independently of each other represent hydrogen or a linear or branched C1-C ₆ alkyl group, or R'd and R'e together with the nitrogen atom they carry form a ring At ₄ , or

R'd, R'e and R'f, together with the nitrogen atom carrying them, form a bridging C ₃ -C ₈ -heterocycloalkyl,

Y1 is a linear or branched C1-C ₄ -alkylene,

Y2 represents the bond, -O-, -O-CH2-, -O-CO-, -O-SO2-, -CH2-, -CH2-O, -CH2-CO-, -CH2-SO2-,

- 5 046336

-C2H5-, -CO-, -CO-O-, -CO-CH2-, -CO-NH-CH2-, -SO2-, -SO2-CH2-, -NH-CO-, -NH-SO2-, m=0, 1 or 2, p=1, 2, 3 or 4,

B1, _B2 , _B3 and _B4 independently represent a _C3 -C8 heterocycloalkyl group which: (i) may be a mono- or bicyclic group, where the bicyclic group includes a fused, bridged or spiro ring system, ( ii) may contain, in addition to the nitrogen atom, one or two heteroatoms independently selected from oxygen, sulfur and nitrogen, (iii) may be substituted by one or two groups selected from: fluorine, bromine, chlorine, linear or branched C ₁ -C ₆ -alkyl, hydroxyl, -NH2, oxo or piperidinyl, it should also be understood that:

aryl means a phenyl, naphthyl, biphenyl or indenyl group, heteroaryl means any mono- or bicyclic group consisting of 5 to 10 ring members, having at least one aromatic moiety and containing from 1 to 4 heteroatoms selected from oxygen, sulfur and nitrogen (including quaternary nitrogen atoms), cycloalkyl means any mono- or bicyclic non-aromatic carbocyclic group containing from 3 to 10 ring members, which may include fused, bridged or spiro ring systems, heterocycloalkyl means any mono- or bicyclic non-aromatic carbocyclic group consisting of 3 -10 ring members, and containing from 1 to 3 heteroatoms selected from oxygen, sulfur, SO, SO2 and nitrogen, it should be understood that the bicyclic group can be of the fused or spiro ring type, heteroarylene, cycloalkylene, heterocycloalkylene means divalent heteroaryl, cycloalkyl and heterocycloalkyl, their enantiomers and diastereoisomers, and their addition salts with a pharmaceutically acceptable acid or base.

Among the pharmaceutically acceptable acids, mention may be made, without implying any limitation, of hydrochloric acid, hydrobromic acid, sulfuric acid, phosphonic acid, acetic acid, trifluoroacetic acid, lactic acid, pyruvic acid, malonic acid, succinic acid, glutaric acid, fumaric acid, tartaric acid, maleic acid, citric acid, ascorbic acid, oxalic acid, methanesulfonic acid and camphoric acid.

Among the pharmaceutically acceptable bases, mention may be made, without implying any limitation, of sodium hydroxide, potassium hydroxide, triethylamine and tert-butylamine.

Further numbered embodiments (E) of the present invention are described below. It should be understood that the features specifically identified in each embodiment may be combined with other specifically identified features to provide additional embodiments of the present invention.

E2. Connection according to option E1, where Het1 is:

E3. A compound according to option E1 or E2, wherein R1 represents a methyl group and R2 represents a hydrogen atom.

E4. A compound according to embodiment E1 or E2, wherein R1 and R2 each represent a methyl group or a hydrogen atom.

E5. A compound according to option E1 or E2, wherein R1 or R2 is a linear or branched ^-^-alkyl, optionally substituted with a hydroxyl or ^-^-alkoxy group, preferably a group selected from methyl, methoxymethyl, hydroxymethyl, ethyl and prop- 2-ila.

E6. A compound according to embodiment E1 or E2, wherein R1 or R2 is a C3-C6 cycloalkyl, preferably cyclopropyl group.

E7. A compound according to option E1 or E2, wherein R1 is a trifluoromethyl group.

E8. A compound according to option E1 or E2, wherein R2 is a linear or branched C ₁ -C ₆ alkylene heterocycloalkyl, preferably (4-methylpiperazin-1yl)propyl group.

E9. A compound according to option E1 or E2, wherein R1 and R2 together with the carbon atoms they bear form a cyclohexene or cyclopentene group.

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E10. A compound according to any one of embodiments E1 to E9, wherein R3 is a hydrogen atom or a methyl group.

E11. A compound according to any one of embodiments E1-E9, wherein R3 is -X1PO(OH)2, -X1-SO2(OH), -X1-NRaRb; -X1-N ⁺ RaRbR _c , where Ra or R _b , or both of them, represent a group selected from C ₁ -C ₆ -alkylene-SO ₂ OH, C ₁ -C ₆ -alkylene-SO ₂ O ^- and C^C^alkylene-PO^H^.

E12. A connection according to any one of embodiments E1 to E9, wherein R8 is -NR' _a R'_b; -N ⁺ R'aR'bR'c;-NH-X'2-N ⁺ R'aR' _b R' _c , where R' _a and R' _b , or both, represent a group selected from C ₁ -C ₆ -alkylene-SO ₂ OH and C ₁ -C ₆ -alkylene-PO(OH) ₂ .

E13. A compound according to any one of embodiments E1 to E9, wherein R3 is a group selected from ethyl, propyl; 2-methoxyethyl, 2-(morpholin-4-yl)ethyl, 3-(morpholin-4-yl)propyl, 3-diethylaminopropyl, 3-methoxypropyl, 3-hydroxypropyl, 3,4-dihydroxybutyl, 4-methoxy-3hydroxybutyl, 4- hydroxy-3-methoxybutyl, 2,3-dihydroxypropyl, 4,5-dihydroxypentyl, 4hydroxybutyl, 3-hydroxy-2-morpholinopropyl, 5-hydroxy-4-methoxypentyl, 5-morpholino-4hydroxypentyl, 3-hydroxy-2-methoxypropyl, 5 - [2-(dimethylamino)ethylmethylamino]-4-hydroxypentyl, 5-hydroxypentyl, 5-methoxy-4-hydroxypentyl, 5-(dimethylamino)-4hydroxypentyl, 4-hydroxy-5-(trimethylammonio)pentyl, 5-[3 -sulfonatepropyldimethylammonio]-4hydroxypentyl, 4-hydroxy-5-(methylamino)pentyl, 3-carboxypropyl, 5-[methyl(4piperidyl)amino]pentyl, 5-(2-morpholinoethylamino)pentyl, 5-[2-(4-methylpiperazine) -1yl)ethylamino]pentyl, 4-[2-(4-methylpiperazin-1-yl)ethylamino]butyl, 4-hydroxy-5-[methyl-[2(methylamino)ethyl]amino]pentyl, 5-[2- (diethylamino)ethylamino]-4-hydroxypentyl, 5-(4-amino-1-piperidyl)-4-hydroxypentyl, 4-hydroxy-5-piperazin-1 -ylpentyl, 5-[2-( 1 -piperidyl)ethylamino]pentyl, 4-(4-amino-1-piperidyl)butyl, 4-[2-(diethylamino)ethylamino]butyl, 5-(4-amino-1piperidyl)pentyl, 4-[methyl-[2-(methylamino)ethyl]amino ]butyl, 4-(2-morpholinoethylamino)butyl, 4[2-(4-methylpiperazin-1-yl)ethylamino]butyl, 4-[(1-methyl-4-piperidyl)amino]butyl, 5-[2( diethylamino)ethylamino]pentyl, 4-piperazin-1-ylbutyl, 4-(methylamino)butyl, 5-piperazin-1ylpentyl, 5-[methyl-2-(methylamino)]ethyl, 5-aminopentyl, 4-aminobutyl, 5- (methylamino)pentyl, 5[3-(4-methylpiperazin-1-yl)propylamino]pentyl, 4-hydroxy-5-[methyl(2-phosphonoethyl)amino]pentyl, 6-(dimethylamino)hexyl, 3-(dimethylamino )propyl, 2-(trimethylammonio)ethyl, 3-(trimethylammonio)propyl, 2-(dimethylamino)ethyl, 4-(dimethylamino)butyl, 5-[3-sulfonatepropyldimethylammonio]pentyl, 4-(trimethylammonio)butyl, 4-hydroxy -5-quinuclidin-1-ium-1-ylpentyl, 4hydroxy-5-(1-aza-4-azoniabicyclo[2.2.2]octan-4-yl)pentyl, 4-hydroxy-5-(4-methylmorpholine-4 -ii-4yl)pentyl, 5-(trimethylammonio)pentyl, 4-hydroxy-5-(1-methylpiperidin-1-ii-1-yl)pentyl, 5-(1,4dimethylpiperazin-1-ii-1-yl) -4-hydroxypentyl, 6-(trimethylammonio)hexyl, 5-[3-hydroxypropyl(dimethyl)ammonio]pentyl, 5-[2-hydroxyethyl(dimethyl)ammonio]pentyl, 5-[carboxymethyl(dimethyl)ammonio]pentyl, 5 -[carboxymethyl(dimethyl)ammonio]pentyl, 3-phosphonopropyl, 3-[3sulfonatepropyldimethylammonio]propyl, 3-sulfopropyl, 4-sulfobutyl, 4-phosphonobutyl, 4carboxybutyl, 3-aminopropyl, 3-azidopropyl, pent-4-in-1 -yl, 4-aminobutyl, 4-azidobutyl, hex5-in-1-yl, 5-azidopentyl.

E14. A compound according to any of embodiments E1-E13, where Het ₂ is: _N coon

-mx 30^ .

E15. A compound according to any of embodiments E1-E13, where Het ₂ is: coon n—(

E16. A compound according to option E14 or E15, wherein R ₆ represents a hydrogen atom.

E17. Connection in accordance with option E14, where R6 represents the group -X ₂ -OR ₇ , where

X ₂ means a propylene group.

E18. Connection according to option E17, where R ₇ represents the following group:

E19. Connection according to option E17, where R ₇ represents the following group:

- 7 046336

E20. Connection according to option E17, where R ₇ represents the following group:

E21. A compound according to any one of embodiments E18-E20, wherein R ₈ is NR'aR'b.

E22. A compound according to any one of embodiments E18-E20, wherein R ₈ is a group selected from dimethylamino, methylamino, methylethylamino, diethylamino, methyl[2-(methylamino)ethyl]amino, (2-hydroxyethyl)(methyl)amino, 4 -morpholinyl, pyrrolidin-1-yl, piperidyl, [(tert-butoxy)carbonyl](methyl)amino, sulfopropylamino, methyl(3-sulfopropyl)amino, methoxyphenyl)methylmethylamino, 2-(dimethylamino)ethylamino and:

hydroxyl, bis(3-sulfopropyl)amino, methyl(p-tolylsulfonyl)amino,

13(4-

E23. A compound according to any one of embodiments E18-E20, wherein R ₈ is a group selected from: 3-piperazin-1-yl, 4-methylpiperazin-1-yl, 3-hydroxypropyl(methyl)amino, [(3S)- 3,4dihydroxybutyl]methylamino, 3-hydroxypropylamino, [(3S)-3,4-dihydroxybutyl]amino, 4hydroxybutyl(methyl)amino, [(3R)-3,4-dihydroxybutyl]methylamino, 4-hydroxybutylamino, [(3R) -3,4dihydroxybutyl]amino.

E24. Connection in accordance with option E17, where R ₇ is:

where Rn is selected from dimethylcarbamoyl, 3-(dimethylamino)propyl, 3-(methylamino)propyl, 3(methylamino)propyl.

E25. A compound according to option E14, where Kb represents a C ₃ -C ₆ heterocycloalkylene^ ₂ ^ ₇ group, where the heterocycloalkylene group is selected from:

E26. A compound according to embodiment E25, wherein R ₇ is selected from methyl, methylethyl, t-butyl, 2-methylpropyl and phenyl, preferably phenyl.

E27. A compound according to option E14 or E15 wherein R ₆ is -C=C(R ₉ )-Y ₁ OR ₇ where Y ₁ is a methylene group.

E28. Connection in accordance with option E14 or E15, where R ₆ is:

E29. Connection according to option E28, where R ₇ is a group selected from:

- 8 046336

where R8 is -O-X'2-NR'aR'b or -X'2-NR'aR'b, preferably -O-X'2-NR'aR'b.

E30. Connection according to option E28, where R ₇ is a group selected from:

where R8 represents a group selected from hydrogen, 2-(methylamino)ethoxy and:

E31. Connection according to option E28, where R ₇ is a group selected from:

where R8 represents a group selected from 2-(dimethylamino)ethoxy, 2-[(2sulfoethyl)amino]ethoxy, 2-[methyl(2-sulfoethyl)amino]ethoxy, 2-(3-hydroxypropylamino)ethoxy, 2(3 -methoxypropylamino)ethoxy, 2-morpholinoethoxy, 2-(2-carboxyethylamino)ethoxy, 2-[(3hydroxyphenyl)methylamino]ethoxy, 2-(methylamino)ethoxy, 2-pyrrolidin-1-ylethoxy, 2-(4methylpiperazine-1- yl)ethoxy, 2-(4-hydroxybutylamino)ethoxy, 2-piperazin-1-ylethoxy, 2-[3hydroxypropyl(methyl)amino]ethoxy, 2-[4-hydroxybutyl(methyl)amino]ethoxy; 2-[[3-hydroxy-2(hydroxymethyl)propyl]amino]ethoxy; 2-[bis(3-hydroxypropyl)amino]ethoxy.

E32. Connection in accordance with option E1, selected from the following group of connections:

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2-({6-[(1,3-Benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(4hydroxybutyl)amino)-5-(3-{2-fluoro-4-[3- (methylamino)prop-1-yn-1yl]phenoxy}propyl)-1,3-thiazol-4-carboxylic acid,

2-[[6-(1,3-Benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]methylamino]-5-[3-[4-[3-(dimethylamino)prop-1-ynyl]- 2fluorophenoxy]propyl]thiazole-4-carboxylic acid,

2-({6-[(1,3-Benzothiazol-2-yl)amino]-5-methylpyridazin-3yl}(methyl)amino)-5-(3-{2-fluoro-4-[3-(methylamino )prop-1-yn-1yl]phenoxy}propyl)-1,3-thiazol-4-carboxylic acid,

2-[[6-(1,3-Benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]-(3,4 dihydroxybutyl)amino]-5-[3-[2-fluoro-4-[3 -(methylamino)prop-1ynyl]phenoxy]propyl]thiazole-4-carboxylic acid,

2-[[6-(1,3-Benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]-(3hydroxy-4-methoxybutyl)amino]-5-[3-[2-fluoro-4-[ 3-(methylamino)prop-1ynyl]phenoxy]propyl]thiazole-4-carboxylic acid,

2-[[6-(1,3-Benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]-(2,3 dihydroxypropyl)amino]-5-[3-[2-fluoro-4-[3 -(methylamino)prop-1ynyl]phenoxy]propyl]thiazole-4-carboxylic acid,

2-[[6-(1,3-Benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]-(4,5 dihydroxypentyl)amino]-5-[3-[2-fluoro-4-[3 -(methylamino)prop-1ynyl]phenoxy]propyl]thiazole-4-carboxylic acid,

- 10 046336

2-[[6-(1,3-Benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]-(3,4dihydroxybutyl)amino]-5-[3-[4-[3-(dimethylamino)prop -1-ynyl]-2fluorophenoxy]propyl]thiazole-4-carboxylic acid,

2-[[6-(1,3-Benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]-(3hydroxypropyl)amino]-5-[3-[2-fluoro-4-[3-(methylamino )prop-1ynyl]phenoxy]propyl]thiazole-4-carboxylic acid,

2-[[6-(1,3-Benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]-[4hydroxy-5-(trimethylammonio)pentyl]amino]-5-[3-[2-fluoro- 4-[3(methylamino)prop-1-ynyl]phenoxy]propyl]thiazole-4-carboxylate,

3-[[5-[[6-(1,3-Benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]-[4carboxy-5-[3-[2-fluoro-4-[3-( methylamino)prop-1-ynyl]phenoxy]propyl]thiazol-2 yl]amino]-2-hydroxypentyl]-dimethylammonio]propane-1-sulfonate,

2-[[6-(1,3-Benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]-(4hydroxybutyl)amino]-5-[3-[2-fluoro-4-[3-[methyl (3-sulfopropyl)amino]prop-1ynyl]phenoxy]propyl]thiazole-4-carboxylic acid,

5-{1-[(Adamantan-1-yl)methyl]-5-methyl-1H-pyrazol-4-yl}-2-({6-[(1,3benzothiazol-2-yl)amino]-5- methylpyridazin-3-yl}(methyl)amino)-1,3-thiazol-4carboxylic acid,

5-{1-[(Adamantan-1-yl)methyl]-5-methyl-1H-pyrazol-4-yl}-2-({6-[(1,3benzothiazol-2-yl)amino]-5- cyclopropylpyridazin-3-yl}(methyl)amino)-1,3thiazol-4-carboxylic acid,

2-[[6-(1,3-Benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]-[5-[2(diethylamino)ethylamino]-4-hydroxypentyl]amino]-5-[3- [4-[3(dimethylamino)prop-1-ynyl]-2-fluorophenoxy]propyl]thiazole-4-carboxylic acid,

2-[[6-(1,3-Benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]-[5-[2-(4methylpiperazin-1-yl)ethylamino] pentyl] amino]-5-[ 3-[4-[3-(dimethylamino)prop 1-ynyl]-2-fluorophenoxy]propyl]thiazole-4-carboxylic acid,

2-[[6-(1,3-Benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]-(4hydroxy-5-piperazin-1-ylpentyl)amino]-5-[3-[4-[ 3-(dimethylamino)prop-1ynyl]-2-fluorophenoxy]propyl]thiazole-4-carboxylic acid,

2-({6-[(1,3-Benzothiazol-2-yl)amino]-5-methylpyridazin-3yl}(methyl)amino)-5-[(28)-3-{4-[3-(dimethylamino )prop-1-in-1-yl]-2fluorophenoxy}-2-methylpropyl]-1,3-thiazol-4-carboxylic acid,

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2-[4-Aminobutyl-[6-(1,3-benzothiazol-2-ylamino)-5-methylpyridazin3-yl]amino]-5-[3-[4-[3-(dimethylamino)prop-1-ynyl ]-2fluorophenoxy]propyl]thiazole-4-carboxylic acid,

2-[5-Aminopentyl-[6-(1,3-benzothiazol-2-ylamino)-5-methylpyridazin3-yl]amino]-5-[3-[4-[3-(dimethylamino)prop-1-ynyl ]-2fluorophenoxy]propyl]thiazole-4-carboxylic acid,

2-[[6-(1,3-Benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]-[3(dimethylamino)propyl]amino]-5-[3-[2-fluoro-4-[ 3-(methylamino)prop-1ynyl]phenoxy]propyl]thiazole-4-carboxylic acid,

2-[[6-(1,3-Benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]methylamino]-5-[3-[4-[3-(dimethylamino)but-1-ynyl]- 2fluorophenoxy]propyl]thiazole-4-carboxylic acid,

2-[[6-(1,3-Benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]-[4hydroxy-5-(trimethylammonio)pentyl]amino]-5-[3-[4-[3 -(dimethylamino)prop-1ynyl]-2-fluorophenoxy]propyl]thiazole-4-carboxylate,

3-[2-[3-[4-[3-[2-[[6-(1,3-Benzothiazol-2-ylamino)-5-methylpyridazin-3yl]-methylamino]-4-carboxythiazol-5-yl ]propoxy]-3-fluorophenyl]prop-2ynylamino]ethyldimethylammonio]propane-1-sulfonate,

2-[[6-(1,3-Benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]-[4hydroxy-5-(4-methylmorpholin-4-ium-4-yl)pentyl]amino]- 5-[3-[4-[3(dimethylamino)prop-1-ynyl]-2-fluorophenoxy]propyl]thiazole-4-carboxylate,

2-[3-[4-[3-[2-[[6-(1,3-Benzothiazol-2-ylamino)-5-methylpyride azin-3-yl]methylamino]-4-carboxythiazol-5-yl] propoxy]-3-fluorophenyl]prop-2ynylamino]ethyltrimethylammonium,

2-[[6-(1,3-Benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]-[6(trimethylammonio)hexyl]amino]-5-[3-[2-fluoro-4-[ 3-(methylamino)prop-1ynyl]phenoxy]propyl]thiazole-4-carboxylate,

2-[[6-(1,3-Benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]-[5-[2hydroxyethyl(dimethyl)ammonio]pentyl]amino]-5-[3-[4- [3-(dimethylamino)prop1-ynyl]-2-fluorophenoxy]propyl]thiazole-4-carboxylate,

2-[[6-(1,3-Benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]-[5[carboxymethyl(dimethyl)ammonio]pentyl]amino]-5-[3-[4-[ 3(dimethylamino)prop-1-ynyl]-2-fluorophenoxy]propyl]thiazole-4-carboxylate,

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3-[[5-[[6-(1,3-Benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]-[4carboxy-5-[3-[2-fluoro-4-[(E) -3-oxoprop-1-enyl]phenoxy]propyl]thiazol-2yl]amino]-2-hydroxypentyl]-dimethylammonio]propane-1-sulfonate,

2-[[6-(1,3-Benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]methylamino]-5-[3-[4-[3-(dimethylamino)prop-1-ynyl]phenoxy ]propyl]thiazole-4carboxylic acid,

2-[[6-(1,3-Benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]methylamino]-5-[3-[4-[1-[(dimethylamino)methyl]-3bicyclo[1.1 .1]pentanyl]phenoxy]propyl]thiazol-4-carboxylic acid, 2-[[6-(1,3-Benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]-(3phosphonopropyl)amino]-5 -[3-[2-fluoro-4-[3-(methylamino)prop-1ynyl]phenoxy]propyl]thiazole-4-carboxylic acid,

2-({6-[(1,3-Benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(3carboxypropyl)amino)-5-(3-{4-[3-(dimethylamino)prop -1-yn-1-yl]-2fluorophenoxy} propyl)-1,3-thiazol-4-carboxylic acid,

2-({6-[(1,3-Benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(3phosphonopropyl)amino)-5-(3-{4-[3-(dimethylamino)prop -1-yn-1-yl]-2fluorophenoxy} propyl)-1,3-thiazol-4-carboxylic acid,

2-({6-[(1,3-Benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(3sulfopropyl)amino)-5-(3-{4-[3-(dimethylamino)prop -1-yn-1-yl]-2fluorophenoxy} propyl)-1,3-thiazol-4-carboxylic acid,

2-({6-[(1,3-Benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(4sulfobutyl)amino)-5-(3-{4-[3-(dimethylamino)prop -1-yn-1-yl]-2fluorophenoxy} propyl)-1,3-thiazol-4-carboxylic acid,

2-({6-[(1,3-Benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(4phosphonobutyl)amino)-5-(3-{4-[3-(dimethylamino)prop -1-yn-1-yl]-2fluorophenoxy}propyl)-1,3-thiazol-4-carboxylic acid,

2-({6-[(1,3-Benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(4carboxybutyl)amino)-5-(3-{4-[3-(dimethylamino)prop -1-yn-1-yl]-2fluorophenoxy}propyl)-1,3-thiazol-4-carboxylic acid,

2-[{6-[(1,3-Benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(pent-4· in-1-yl)amino]-5-(3-{2 -fluoro-4-[3-(methylamino)prop-1-yn-1yl]phenoxy}propyl)-1,3-thiazole-4-carboxylic acid,

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2-({6-[(1,3-Benzothiazol-2-yl)amino]-5-methylpyridazin-3yl}(methyl)amino)-5-{3-[2-fluoro-4-(3-{methyl [2-(methylamino)ethyl]amino}prop-1in-1-yl)phenoxy]propyl}-1,3-thiazol-4-carboxylic acid,

2-({6-[(1,3-Benzothiazol-2-yl)amino]-5-methylpyridazin-3yl}(methyl)amino)-5-[3-(2-fluoro-4-{3-[( 2-hydroxyethyl)(methyl)amino]prop-1-yn1-yl}phenoxy)propyl]-1,3-thiazol-4-carboxylic acid,

2-({6-[(1,3-Benzothiazol-2-yl)amino]-5-methylpyridazin-3yl}(methyl)amino)-5-{3-[2-fluoro-4-(3-hydroxyprop- 1-yn-1-yl)phenoxy]propyl}1,3-thiazol-4-carboxylic acid,

2-[5-Azidopentyl-[6-[(1,3-benzothiazol-2-yl)amino]-5methylpyridazin-3-yl]amino]-5-[3-[2-fluoro-4-[3-( methylamino)prop-1ynyl]phenoxy]propyl]-1,3-thiazole-4-carboxylic acid, its enantiomers and diastereoisomers, and their addition salts with a pharmaceutically acceptable acid or base.

E33. A compound according to option E1, selected from the following group of compounds: 6-[{6-[(1,3-Benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl} (methyl)amino] -3 - [ 1 -({3 -[2-(dimethylamino)ethoxy]-5,7-dimethyladamantan-1 yl}methyl)-5-methyl-1H-pyrazol-4-yl]pyridin-2-carboxylic acid,

6-[[6-(1,3-Benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]methylamino]-3 -[ 1 -[[3 -[2-(3-hydroxypropylamino)ethoxy]-5 ,7-dimethyl-1 adamantyl]methyl]-5-methylpyrazol-4-yl]pyridine-2-carboxylic acid,

6-[[6-(1,3-Benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]methylamino] -3 -[ 1 -[[3 - [2-(3-methoxypropylamino)ethoxy]-5 ,7-dimethyl-1 adamantyl]methyl]-5-methylpyrazol-4-yl]pyridin-2-carboxylic acid, 6-[{6-[(1,3-Benzothiazol-2-yl)amino]-5-methylpyridazine -3yl}(methyl)amino]-3-(1-{[3-(2-{[(3 8)-3,4-dihydroxybutyl]amino}ethoxy)-5,7dimethylad amantan-1-yl]methyl} -5-methyl-1H-pyrazol-4-yl)pyridin-2-carboxylic acid,

6-[[6-(1,3-Benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]methylamino]-3-[1-[[3,5-dimethyl-7-(2-morpholinoethoxy)- 1-adamantyl]methyl]-5methylpyrazol-4-yl]pyridin-2-carboxylic acid, its enantiomers and diastereoisomers, and their addition salts with a pharmaceutically acceptable acid or base.

E34. A method for producing a compound of formula (I) in accordance with any of variants E1-E33, which is characterized in that a compound of formula (II) is used as a starting material:

wherein R1 and R2 are as defined for formula (I), wherein said compound of formula (II) is subjected to a Buchwald reaction in an aqueous or organic medium in the presence of a palladium catalyst (preferably Pd2(dba)3), a base (preferably Cs2CO3 and DIPEA) , phosphine (preferably xanthos) and a compound of formula (III):

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6-[[6-(1,3-Benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]methylamino]-3-[1-[[3-[2-(2-carboxyethylamino)ethoxy]-5 ,7-dimethyl-1adamantyl]methyl]-5-methylpyrazol-4-yl]pyridin-2-carboxylic acid, 6-[[6-(1,3-Benzothiazol-2-ylamino)-5-methylpyridazin-3-yl ]methylamino]-3-[1-[[3-[2-[(3-hydroxyphenyl)methylamino]ethoxy]-5,7-dimethyl-1adamantyl]methyl]-5-methylpyrazol-4-yl]pyridin-2- carboxylic acid, 2-[[6-(1,3-Benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]methylamino]-5-[3-[4-[3-(dimethylamino)prop-1- inyl]-2fluorophenoxy]propyl]thiazole-4-carboxylic acid,

2-[[6-(1,3-Benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]-(3hydroxypropyl)amino]-5-[3-[2-fluoro-4-[3-(methylamino )prop-1ynyl]phenoxy]propyl]thiazole-4-carboxylic acid,

2-[[6-(1,3-Benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]methylamino]-5-[3-[2-fluoro-4-(3-pyrrolidin-1-ylprop- 1ynyl)phenoxy]propyl]thiazole-4-carboxylic acid, where R4 and m are as defined for formula (I), to give a compound of formula (IV):

the amine function of the compound of formula (IV) is further protected to give the compound of formula (V):

where _k PG represents a protecting group (preferably 2trimethylsilylethoxymethyl group), the compound of formula (V) is then subjected to a Buchwald reaction in an aqueous or organic medium in the presence of a palladium catalyst (preferably Pd2(dba)3), a base (preferably Cs2CO3 and DIPEA), phosphine ( preferably xanthos) and compounds of formula (VI):

where A1, R3 and R6 are as defined for formula (I), and Alk represents a C1-C ₆ alkyl group, to obtain a compound of formula (VII-a) or (VII-b):

- 15 046336 and then the benzothiazole group of said compound of formula (VII-a) or (VII-b) is deprotected (preferably using HFxPyr) and the ester function is hydrolyzed (preferably using LiOHxH2O) to obtain a compound of formula (I), wherein the compound of formula (I) may be purified according to conventional resolution techniques, may be converted into its addition salts with a pharmaceutically acceptable acid or base, and optionally resolved into its isomers according to conventional separation techniques, it being understood that, at any time deemed appropriate during the course of the described above the method, the hydroxy, amino, carboxyl and phosphono groups of the reagents or intermediates of the synthesis can be protected and then deprotected as required by the synthesis.

E35. Method according to option E34, wherein compound (VI-b) is:

where R8, R ₁₂ and R ₁₃ are as defined for formula (I).

E36. Process for preparing a compound of formula (I) according to embodiment E28, wherein R ₆ is:

and R ₇ is as defined for formula (I), which is characterized in that a compound of formula (II) is used as a starting material:

wherein R ₁ and R ₂ are as defined for formula (I), wherein the compound of formula (II) is subjected to a Buchwald reaction in an aqueous or organic medium in the presence of a palladium catalyst (preferably Pd ₂ (dba) ₃ ), a base (preferably Cs ₂ CO ₃ and DIPEA), phosphine (preferably xanthos) and a compound of formula (III):

where R ₄ and m are as defined for formula (I), giving a compound of formula (IV):

the amine function of the compound of formula (IV) is further protected to give the compound of formula (V):

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(pg) where represents a protecting group (preferably a 2trimethylsilylethoxymethyl group), the compound of formula (V) is then subjected to a Buchwald reaction in an aqueous or organic medium in the presence of a palladium catalyst (preferably Pd ₂ (dba) ₃ ), a base (preferably Cs ₂ CO ₃ and DIPEA), phosphine (preferably xanthos) and a compound of formula (VI'):

where A1 and R3 are as defined for formula (I), and Alk represents a C1-C6 alkyl group, giving a compound of formula (VIII):

a compound of formula (VIII) is halogenated (preferably using Nbromosuccinimide or N-iodosuccinimide) to give a compound of formula (IX):

where Hal represents a halogen atom, the compound of formula (IX) is then subjected to a Suzuki coupling reaction, in an aqueous or organic medium, in the presence of a palladium catalyst (preferably Pd(dppf)Cl2), a base (preferably potassium carbonate), with a compound of formula (X):

to obtain a compound of formula (XI):

and then the benzothiazole group of said compound of formula (XI) is deprotected (preferably using HFxPyr) and the ester function is hydrolyzed (preferably using LiOHxH2O) to obtain the compound of formula (I), wherein the compound of formula (I) can be purified according to conventional separation techniques, can be converted in its addition salt with a pharmaceutically acceptable acid or base and optionally separated into its isomers according to conventional separation techniques, it being understood that, at any time deemed appropriate during the course of the above

- 17 046336 method, the hydroxy, amino, carboxyl and phosphono groups of the reagents or synthesis intermediates can be protected and then deprotected as required by the synthesis.

E37. A synthesis intermediate according to option E34 or E36, which is:

where R ₄ and m are as defined for general formula (I), preferably R ₄ represents a hydrogen atom.

E38. A compound according to option E1, wherein R4 is hydrogen, a fluorine, chlorine or bromine atom, a methyl or methoxy group.

E39. A compound according to embodiment E1, wherein R ₈ is a group selected from hydrogen; linear or branched ^-^-alkyl, -NR'aR'b;-NR'a-CO-OR'c; -N ⁺ R' _a R' _b R'_c;-O-R'_c; -NH- +

X'2-N+R'aR'bR'c; -O-X'2-NR'aR'b, -NR'c-X'2-Nj and .

E40. The compound according to E1, wherein R'a and _R'b independently represent a group selected from hydrogen; heterocycloalkyl; -SO ₂ ^h^^ where phenyl may be replaced by linear or branched ^-^-alkyl; linear or branched ^-^-alkyl, optionally substituted with one or two hydroxyl groups; C ₁ -C ₆ -alkylene-SO ₂ OH; C ₁ -C ₆ -αalkylene-SO ₂ O ^- ; C1-C6-alkylene-COOH; C1-C6-αalkylene-PO(OH)2; C1-C6-alkylene-NR'dR'e; C1-C6-alkylene-N ⁺ R'dR' _e R'_f; C ₁ -C ₆ αalkylene-O-C ₁ -C ₆ -alkylene-OH; ^-^-alkylenephenyl, where phenyl may be replaced by a ^-^-alkoxy group;

groups:

or R'a and R'b, together with the nitrogen atom they carry, form a B ₃ ring, or R'a, R'b, and _R'c, together with the nitrogen atom they carry, form a bridged C ₃ -C ₈ -heterocycloalkyl.

E41. Connection in accordance with any of options E1-E31, where m=1.

Pharmacological study of the compounds of the present invention showed that they have pro-apoptotic properties. The ability to reactivate the apoptotic process in cells is of great therapeutic interest for the treatment of malignant neoplasms and immunopathological and autoimmune diseases. In particular, the compounds of the invention will be useful for the treatment of chemo- or radioresistant malignancies.

In another embodiment, the compounds of the present invention can be used to treat diseases or conditions characterized by excessive or deregulated platelet activity, especially prothrombotic conditions.

As used herein, the term treat or cure any disease or disorder refers in one embodiment to reducing the severity of the disease or disorder (ie, slowing or stopping or reducing the progression of the disease or at least one of its clinical symptoms). In another embodiment, the term treat or treat refers to the mitigation or reduction in intensity of at least one physical parameter, including those that may not be felt by the patient. In yet another embodiment, the term treat or treatment refers to modulating a disease or disorder, either physically (eg, stabilization of a tangible symptom), physiologically (eg, stabilization of a physical parameter), or both.

Among the cancer treatment options contemplated herein may be mentioned, without implying any limitation, the treatment of hematologic malignancies and solid tumors. Hematologic malignancies include myeloma, especially multiple myeloma, lymphoma, especially non-Hodgkin's lymphoma (NHL) and more specifically diffuse large B-cell lymphoma (DLBCL), and leukemia, especially chronic lymphocytic leukemia (CLL), T-cell acute lymphoblastic leukemia (T-ALL), B-cell acute lymphoblastic leukemia (B-ALL) and acute myelocytic leukemia (AML). Solid tumors include bladder, brain, breast, uterine, esophageal and liver cancer, colorectal cancer, kidney cancer, melanoma, ovarian cancer, prostate cancer, pancreatic cancer and lung cancer, especially non-small cell lung cancer and small cell lung cancer .

In particular, T-ALL arises as a result of leukemic transformation of cell precursors

- 18 046336 thymus current and their blocking at certain stages of differentiation. Despite valuable information recently obtained from extensive research into the molecular and cellular mechanisms responsible for the occurrence and progression of T-ALL, this knowledge has not been translated into effective targeted therapies. Current clinical treatments include chemotherapy, with or without hematopoietic stem cell transplantation, with survival rates remaining around 50 and 70% in adults and children, respectively. In both children and adults, relapses demonstrate a very poor prognosis, increasing the need to identify new therapeutic treatments (Passaro et al., Immunol. Rev. 2016 Mau; 271(1):156–72). Dual Bcl-2/Bcl-xL inhibitors like ABT-263 and ABT-737 have been shown to have promising activity in xenograft models derived from patients with TALL (Van Delft et al. Cancer Cell 2006; 10:389-99 ; Suryani et al., Clin. Cancer Res. 2014, 20:4520-31). Other studies have reported a differential requirement for Bcl-xL or Bcl-2 for survival of mature versus very immature (ETP subgroup) T-ALL (Chonghaile et al., Cancer Discov. 2014;4:1074-87). The previously described selective Bcl-xL inhibitor A-1331852 has also been shown to have in vitro and in vivo activity in a xenograft model of the mature T-ALL cell line MOLT4 (Leverson et al., Sci. Transl. Med. 2015 Mar 18;7 (279):279ra40). In a separate embodiment, inhibition of tumor growth was also observed in the MOLT-4 xenograft model following treatment with the Bcl-xL inhibitors of the present invention. These data support the use of the compounds of the present invention for the treatment of T-ALL.

Among the treatment options for autoimmune diseases contemplated herein are, without limitation, the treatment of rheumatoid arthritis (RA) and systemic lupus erythematosus (SLE).

The present invention also relates to pharmaceutical compositions containing at least one compound of formula (I), as an active ingredient, in combination with one or more pharmaceutically acceptable excipients. In particular, such pharmaceutical compositions are interesting for use as proapoptotic and/or antiproliferative agents, in particular for the treatment of malignancies, as well as autoimmune diseases and diseases of the immune system.

Suitable excipients according to the invention include diluents, lubricants, binders, disintegrants, stabilizers, preservatives, absorbents, coloring agents, sweeteners and flavoring agents.

By way of non-limiting example the following may be mentioned:

as diluents: lactose, dextrose, sucrose, mannitol, sorbitol, cellulose, glycerin, as lubricants: silicon dioxide, talc, stearic acid and its magnesium and calcium salts, polyethylene glycol, as binders: magnesium aluminum silicate, starch , gelatin, tragacanth, methylcellulose, sodium carboxymethylcellulose and polyvinylpyrrolidone, as raising agents: agar, alginic acid and its sodium salt, effervescent mixtures.

Among the pharmaceutical compositions according to the invention, mention may be made more particularly of those suitable for oral, parenteral, nasal, trans- or transdermal, rectal, perlingual, ophthalmic or respiratory administration, in particular tablets, dragees, sublingual tablets, capsules, glossets, capsules, lozenges, injectable or drinkable preparations, aerosols, eye or nasal drops, suppositories, creams, ointments, skin gels.

The actual dosage levels of the active ingredients in the pharmaceutical compositions of the present invention may vary to provide an amount of the active ingredient that is effective to achieve the target therapeutic response for a particular patient, composition, and route of administration, but is not toxic to the patient. The dosage level chosen will depend on a number of factors, including the potency of the particular compound used in the present invention, route of administration, time of administration, rate of elimination or metabolism of the particular compound used, rate and extent of absorption, duration of treatment, other drugs, compounds and/or substances agents used in combination with the particular compound used, the age, sex, weight, condition, general health and previous medical history of the patient being treated, and similar factors well known in the medical field.

A suitable daily dosage of a compound of the present invention will depend on the factors described above and may vary from 0.01 mg to 2.5 g per day in one or more administrations.

In another aspect, the present invention also relates to the combination of a compound of formula (I) with an antitumor agent selected from genotoxic agents, mitotic poisons, antimetabolites, proteasome inhibitors, kinase inhibitors and antibodies, as well as pharmaceutical compositions containing such type of combination, and their use for the manufacture of medicinal products for use in the treatment of malignant neoplasms.

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In another aspect, the compounds of the present invention can be used in combination with radiotherapy for the treatment of cancer.

Alternatively, the compounds of the present invention may be associated with monoclonal antibodies. Antibody-drug conjugates (ADCs) are a new class of therapeutics that are formed by chemically linking a cytotoxic agent to a monoclonal antibody through a linker. The monoclonal antibody from the ADC selectively binds to the target antigen of a cell (such as a cancer cell) and releases the drug into the cell. ADCs have therapeutic potential because they combine the specificity of an antibody with the cytotoxic potential of a drug. However, the development of ADCs as therapeutic agents has so far only had limited success due to multiple factors, such as an unfavorable toxicity profile, low efficacy, and poor pharmacological parameters. Accordingly, there is still a need for new ADCs that overcome these problems and can selectively deliver Bcl-xL to targeted cancer cells.

In another aspect, the compounds of the present invention may be linked to monoclonal antibody fragments or linked to scaffold proteins, which may or may not be monoclonal antibodies. Antibody fragments should be understood as fragments of Fv, scFv, Fab, F(ab')2, F(ab'), scFv-Fc type or diabody, which generally have the same binding specificity as the antibody from which they are derived. In accordance with the present invention, antibody fragments of the present invention can be obtained from antibodies using methods such as digestion with enzymes such as pepsin or papain, and/or through cleavage of disulfide bridges using chemical reduction. Alternatively, fragments of the antibodies included in the present invention can be obtained using genetic recombination techniques, also well known to one skilled in the art, or even through peptide synthesis using, for example, automated peptide synthesizers such as those available from Applied Biosystems, etc.

By scaffold proteins, which may or may not be monoclonal antibodies, is meant a protein that may or may not contain an immunoglobulin chain fold and that provides the same binding ability as a monoclonal antibody. One skilled in the art will know how to select a protein scaffold. More specifically, it is known that a framework must be selected that exhibits several of the following distinguishing features (Skerra, J. Mol. Recogn. 2000, 13, 167-187): good phylogenetic conservation, robust architecture with a well-known three-dimensional molecular organization ( as, for example, based on crystallography or NMR), small size, absence or only a low degree of post-translational modifications, ease of preparation, expression and purification. Such a scaffold protein may be, but is not limited to, a structure selected from the group consisting of fibronectin and preferably the tenth domain of fibronectin type III (FNfn10), lipocalin, anticalin (Skerra, J. Biotechnol. 2001, 74, 257-75), protein Z, which is derived from the B domain of staphylococcal protein A, thioredoxin A, or any protein with a repeat domain such as an ankyrin repeat (Kohl et al. PNAS 2003, 100, 1700-1705), armadillo repeat, leucine-rich repeat, or tetratricopeptide repeat. Mention may also be made of a framework derived from toxins (such as scorpion, insect, plant or shellfish toxins) or protein inhibitors of neuronal nitric oxide synthase (PIN).

The following examples illustrate the invention but do not limit it in any way. All intermediates for preparing the compounds of the examples are either commercially available or can be prepared by one skilled in the art using conventional chemical reactions described in the literature.

General techniques

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

Column chromatography.

Automated flash column chromatography was performed on an ISCO CombiFlash® Rf 200 or CombiFlash® Rf+ Lumen™ instrument using RediSep® Rf flash chromatography columns on normal phase silica gel (35-70 μm, 60 A), RediSep Rf Gold® columns for normal phase silica gel high performance chromatography (20-40 µm, 60 A), C18 RediSep® Rf reverse phase chromatography column (40-63 µm, 60 A) or C18 RediSep Rf Gold® reverse phase high performance chromatography column (20- 40 µm, 100 A).

TLC.

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

Reactions in a microwave reactor.

- 20 046336

Microwave heating was performed in a CEM Discove® SP or an Anton Paar single-mode microwave reactor.

NMR.

1H-NMR measurements were performed on a Bruker Avance III 500 MHz spectrometer, a Bruker Avance III 400 MHz spectrometer or a Bruker DPX-400 spectrometer using DMSO-dg or CDCl ₃ as solvent. 1H NMR data are presented as delta values reported in parts per million (ppm) using the residual solvent peak (2.50 ppm for DMSO-dg and 7.26 ppm for CDCl ₃ ) as the internal standard. The splitting patterns are designated as: s (singlet), d (doublet), t (triplet), q (quartet), quint (quintet), sept (septet), m (multiplet), br s (wide singlet), dd ( doublet of doublets), td (triplet of doublets), dt (doublet of triplets), ddd (doublet of doublet of doublets).

Analytical LC-MS.

Certain compounds of the present invention were characterized by high performance liquid chromatography-mass spectroscopy (HPLC-MS) on an Agilent HP1200 instrument with an Agilent 6140 quadrupole LC/MS operating in electrospray ionization in positive or negative ion mode. The molecular weight scanning range was from 100 to 1350. Parallel UV detection was performed at wavelengths of 210 nm and 254 nm. Samples were injected as a 1 mM solution in ACN or a THF/ _H2O mixture (1:1) in a volume of 5 μl using a loop pipette. LCMS analyzes were performed on two instruments, one of which worked with basic and the other with acidic eluents.

Basic LCMS: Gemini-NX, 3 µm, C18, 50 mm x 3.00 mm i.d. column at 23°C, at a flow rate of 1 ml min ^-1 , using 5 mM ammonium bicarbonate (solvent A) and acetonitrile (solvent B) with a gradient starting with 100% solvent A and ending with 100% solvent B over various periods. certain periods of time.

Acidic LCMS: KINATEX XB-C18-100A, 2.6 µm, 50 mm x 2.1 mm column at 40°C, at a flow rate of 1 ml min ^-1 , using 0.02% v/v. aqueous solution of formic acid (solvent A) and 0.02% v/v. a solution of formic acid in acetonitrile (solvent B) with a gradient starting with 100% solvent A and ending with 100% solvent B over various/defined periods of time.

Certain other compounds of the present invention have been characterized by HPLC-MS using the specifically named methods as set forth below. For all these methods, UV detection was performed using a diode array detector at wavelengths of 230, 254, and 270 nm. The injected sample volume was 1 μL. Gradient elution operations were carried out by adjusting the flow rates and percentages of the mixtures of the following mobile phases using HPLC grade solvents:

Solvent A: 10 mM aqueous ammonium formate + 0.04% (v/v) formic acid.

Solvent B: acetonitrile + 5.3% (v/v) solvent A + 0.04% (v/v) formic acid.

The retention time (RT) values for these named methods are given in minutes. Ionization was recorded in the detection mode of positive ions, negative ions, or in the mode with switching detection of positive/negative ions. Specific details for individual methods are provided below.

LCMS-V-B methods.

Used an Agilent 1200 SL Series instrument coupled to an Agilent MSD 6140 single quadrupole detector with a multimode ESI-CIAD source (LCMS-V-BI and LCMSL'-B2 methods), or used an Agilent 1290 Infinity II Series instrument coupled to an Agilent TOF 6230 with ERJet Stream source (LCMS-V-BI method); column: Thermo Accucore 2.6 µm, C18, 50 mm x 2.1 mm at 55°C. Detailed data regarding the gradient for the LCMS-V-BI and Zh ^- X\1SL'-B2 methods:

Time (min) LCMS-V-B 1 ZhKhMS-U-V2 Flow (ml/min) Solvent A (%) Solvent B (%) Solvent A (%) Solvent B (%) 0 95 5 60 40 1.1 0.12 95 5 60 40 1.3 1.30 5 95 2 98 1.3 1.35 5 95 2 98 1.6 1.85 5 95 2 98 1.6 1.90 5 95 2 98 1.3 1.95 95 5 95 5 1.3

- 21 046336

LCMS-V-C method.

An Agilent 1200 SL series instrument was used, coupled with an Agilent MSD 6140 single quadrupole detector with a multimode ESI-CIAD source; Column: Agilent Zorbax Eclipse plus 3.5 µm, C18(2), 30 mm x 2.1 mm at 35°C. Gradient details for the LCMS-V-C method:

Time (min) Solvent A (%) Solvent B (%) Flow(ml/min) 0 95 5 1 0.25 95 5 1 2.50 95 5 1 2.55 5 95 1.7 3.60 5 95 1.7 3.65 5 95 1 3.70 95 5 1 3.75 95 5 1

Preparative HPLC.

Certain compounds of the present invention were purified by high performance liquid chromatography (HPLC) on an Armen Spot Liquid Chromatography or Teledyne EZ system with Gemini-NX® 10 µm C18, 250 mm x 50 mm i.d. column, operating at a flow rate of 118 ml min ^-1 with a diode array UV detector (210-400 nm), and using a 25 mM aqueous solution of NH ₄ HCO ₃ and MeCN or 0.1% TFA in water and MeCN as eluents.

Certain other compounds of the present invention were purified by HPLC using the specifically named methods as set forth below.

HPLC-V-A methods.

Operations were performed on a Waters FractionLynx MS autocleaning system, with Gemini® 5 µm C 18(2), 100 mm x 20 mm i.d. column from Phenomenex, operating at a flow rate of 20 cm ³ min ^-1 with a UV diode array detector (210-400 nm) and mass-directed collection. The mass spectrometer was a Waters Micromass ZQ2000 electrospray ionization spectrometer in positive or negative ion mode with a molecular weight scan range of 150 to 1000.

HPLC-V-AI method (pH 4):

Solvent A: 10 mM aqueous ammonium acetate + 0.08% (v/v) formic acid; Solvent B: acetonitrile + 5% (v/v) solvent A + 0.08% (v/v) formic acid.

HPLC-V-A method (pH 9):

Solvent A: 10 mM aqueous ammonium acetate + 0.08% (v/v) conc. ammonia solution; Solvent B: Acetonitrile + 5% (v/v) solvent A + 0.08% (v/v) conc. ammonia solution.

HPLC-V-B methods.

Surgeries were performed on the AccQPrep HP125 system (Teledyne ISCO) with Gemini® NX 5 µm C 18(2), 150 mm x 21.2 mm i.d. column from Phenomenex, operating at a flow rate of 20 cm ³ min ^-1 with detection on a UV detector (214 and 254 nm) and IDSR.

HPLC-V-BI method (pH 4):

Solvent A: water + 0.08% (v/v) formic acid; Solvent B: acetonitrile + 0.08% (v/v) formic acid.

HPLC-U-B2 method (pH 9):

Solvent A: water + 0.08% (v/v) conc. ammonia solution; Solvent B: Acetonitrile + 0.08% (v/v) conc. ammonia solution.

HPLC-\'-VZ method (neutral):

Solvent A: water; Solvent B: acetonitrile.

Analytical GC-MS.

Combined gas chromatography-low-resolution mass spectrometry (GC-MS) was performed on an Agilent 6850 gas chromatograph and an Agilent 5975C mass spectrometer using a 15 m x 0.25 mm column with a 0.25 μm HP-5MS coating and helium as the carrier gas. Ion source: EI ⁺ , 70 eV, 230°C, quadrupole: 150°C, interface: 300°C.

High resolution MS.

High-resolution mass spectra were obtained on an Agilent 6230 time-of-flight mass spectrometer equipped with a Jet Stream electrospray ion source operating in positive ion mode. An injection volume of 0.5 µl was sent to the mass spectrometer at a flow rate of 1.5

- 22046336 ml/min (gradient program: 5 mM ammonium formate in water and acetonitrile), using an Agilent 1290 Infinity HPLC system. Jet Stream parameters: flow rate and temperature of drying gas (N ₂ ): 8.0 l/min and 325°C, respectively; atomizing gas (N2) pressure: 30 psi. inch; capillary voltage: 3000 V; flow rate and temperature of protective gas: 325°C and 10.0 l/min; TOFMS parameters: fragmentor voltage: 100 V; skimmer potential: 60 V; OST 1 RF Vpp: 750 V. Full scan mass spectra were obtained in the range m/z 105-1700 at an acquisition rate of 995.6 ms/spectrum and processed using Agilent MassHunter B.04.00 software.

Name of chemical compounds.

Preferred IUPAC names were generated using ChemAxon's 'Structure to Name' (s2n) function within MarvinSketch or JChem for Excel (JChem versions 16.6.13 - 18.22.3), or using the chemical name generation function provided by Biovia® Draw 4.2.

Abbreviations.

Ahx - 6-hexanoic acid monomer;

Boc - tert-butyloxycarbonyl;

Boc ₂ O - di-tert-butyl dicarbonate;

tBuOH - tert-butanol;

conc. - concentrated;

CuOH - cyclohexanol;

dba - (1E,4E)-1,5-diphenylpenta-1,4-dien-3-one, dibenzylideneacetone;

DCM - dichloromethane;

DEA - diethanolamine;

DIAD - diisopropyl azodicarboxylate;

DIBAL-H - diisobutylaluminum hydride;

DIPA - N-isopropylpropan-2-amine, diisopropylamine;

DIPEA - Nethyl-N-isopropylpropan-2-amine, diisopropylethylamine;

EtO2 - diethyl ether;

EtOAc - ethyl acetate;

eq. - equivalent;

HFxPyr - hydrogen fluoride-pyridine;

hs - homo sapiens;

LDA - lithium diisopropylamide;

MeCN - acetonitrile;

MeOH - methanol;

MTBE - methyl tert-butyl ether;

VN - during the night;

Pd\C - palladium on carbon;

PPh ₃ - triphenylphosphine;

k.t. - room temperature;

RT - retention time (in minutes);

TBAF - tetrabutylammonium fluoride;

TBAON - tetrabutylammonium hydroxide;

TBSCl - tert-butylchlorodimethylsilane;

TEA - K^diethylethanamine;

TFA - 2,2,2-trifluoroacetic acid;

pTSA - 4-methylbenzenesulfonic acid;

THF - tetrahydrofuran;

TIPSCl - chloro(triisopropyl)silane;

xantphos - 4,5-bis(diphenylphosphino)-9,9-dimethylxanthene;

xantphos Pd G3 - methanesulfonate [(4,5-bis(diphenylphosphino)-9,9-dimethylxanthene)-2-(2'-amino1,1'-biphenyl)] palladium(P);

BINAP - 2,2'-bis(diphenylphosphino)-1,1'-binaphthyl;

rac-BINAP Pd G3 - methanesulfonate [(2,2'-bis(diphenylphosphino)-1,1'-binaphthyl)-2-(2'-amino-1,1'biphenyl)] palladium(P);

Pd(dppf)Cl ₂ - CH ₂ Cl ₂ [1,G-bis(diphenylphosphino)ferrocene]dichloropalladium(P);

Pd ₂ (dba) ₃ - tris(dibenzylideneacetone)dipalladium(0);

Pd(PPh ₃ ) ₂ Cl ₂ - bis(triphenylphosphine) palladium chloride;

Pd(AtaPhos) ₂ Cl ₂ - bis(di-tert-butyl(4-dimethylaminophenyl)phosphine)dichloropalladium(P) Methods with a well-known name.

Below are representative experimental procedures that are referred to by name in the following Syntheses.

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General procedure for carrying out the Sonogashira reaction.

Mixture 1 eq. aryl halide, 2 eq. acetylene, 0.05 equiv. Pd(PPh ₃ ) ₂ Cl ₂ , 0.05 eq. CuI and DIPA (1 ml/mmol) in THF (5 ml/mmol) were kept at 60°C. Once a suitable conversion was achieved, volatiles were removed under reduced pressure and the crude intermediate was purified using flash column chromatography.

General procedure for removing protection using HFIP.

The substrate in 1,1,1,3,3,3-hexafluoropropan-2-ol (10 ml/mmol) was maintained at 100-120°C in a pressure reaction vessel. Once a suitable conversion was achieved, volatiles were removed under reduced pressure and the crude intermediate was purified using flash column chromatography.

General alkylation procedure.

Mixture 1 eq. phenol/carbamate, 1-2 eq. alkyl iodide or bromide and 2-3 eq. Cs ₂ CO ₃ in acetone (5 ml/mmol) was stirred at room temperature. in the case of phenols and at 55°C in the case of carbamates. Once a suitable conversion was achieved, volatiles were removed under reduced pressure and the crude intermediate was purified using flash column chromatography or reverse phase flash column chromatography.

General procedure for performing the Mitsunobu reaction.

To the mixture 1.0-1.5 eq. aliphatic alcohol, 1 eq. carbamate/phenol and 1-2 eq. 1-3 eq. of triphenylphosphine in THF or toluene (5 ml/mmol) was added in one portion. di-tert-butyl azodicarboxylate/diisopropyl azodicarboxylate. The mixture was stirred at room temperature. or, if necessary, 50°C in the case of carbamate, and at k.t. in the case of phenol. Once a suitable conversion was achieved, volatiles were removed under reduced pressure and the crude intermediate was purified using flash column chromatography.

General procedure I for carrying out the Buchwald reaction.

Mixture 1 eq. chlorine-containing substrate, 2 eq. 1,3-benzothiazol-2-amine, 0.1 eq. Pd ₂ (dba) ₃ , 0.2 eq. XantPhos and 3 eq. DIPEA in CyOH (5 ml/mmol) was kept at 140°C. Once a suitable conversion was achieved, the reaction mixture was diluted with DCM (10 ml/mmol), applied to a preconditioned silica gel column and purified by flash column chromatography.

General procedure II for carrying out the Buchwald reaction.

Mixture 1 eq. thiazolamine, 1.2-1.5 eq. (2)-N(6-chloro-4-methylpyridazin-3-yl)-3-(2trimethylsilylethoxymethyl)-1,3-benzothiazol-2-imine, 3 eq. Cs ₂ CO ₃ , 0.1 eq. Pd ₂ (dba) ₃ , 0.2 eq. XantPhos and 3 eq. DIPEA in 1,4-dioxane (5 ml/mmol) was kept under reflux. Once a suitable conversion was achieved, volatiles were removed under reduced pressure and the crude intermediate was purified using flash column chromatography.

General procedure for the Finkelstein reaction.

Mixture 1 eq. alkyl chloride and 2 eq. NaI in acetone (5 ml/mmol) was kept under reflux. Once a suitable conversion was achieved, volatiles were removed under reduced pressure and the crude intermediate was purified by flash column chromatography.

General procedure for carrying out nucleophilic substitution reactions.

Mixture 1 eq. thiazolamine, 1 eq. 3,6-dichloro-4-methylpyridazine and 1.5 eq. Cs ₂ CO ₃ in toluene (4 ml/mmol) was kept at 150°C. Once a suitable conversion was achieved, volatiles were removed under reduced pressure and the crude intermediate was purified by flash column chromatography.

General procedure for tosylate alkylation.

The oven-dried vessel was equipped with a PTFE-coated magnetic stirrer armature, and 1 eq. tosylate and 5 eq. suitable amine suspended in MeCN (5 ml/mmol). The reaction mixture was then heated to 50°C and stirred at this temperature until no further transformation was observed. The reaction mixture was diluted with DCM and then applied to a DCM-preconditioned silica gel column. The crude product was then purified by flash column chromatography using DCM and MeOH (1.2% NH3) as eluents.

General procedure for alkylation with in situ formed tosylate.

The oven-dried vessel was equipped with a PTFE-coated magnetic stirrer armature, and 1 eq. hydroxy derivative, 3 eq. N.N-diethylethanamine. catalytic amount of N-dimethylpyridin-4-amine and DCM (5 ml/mmol), then 2 eq. was added in one portion. 4-methylbenzenesulfonyl chloride, and the mixture was stirred at room temperature. until no further transformation occurs (typically >90%, 1-5 hours). After evaporation of volatiles at 40°C using N2, 30 eq. suitable primary, secondary or tertiary amine in acetonitrile (3 ml/mmol), and the mixture was stirred at 50°C until complete conversion of the tosyl derivative was observed (typically 1-3 hours). The reaction mixture was concentrated

- 24 046336 on celite and purified by flash column chromatography using EtOAc and MeOH (1.2% NH ₃ ) as eluents, or by reverse phase flash column chromatography (0.1% TFA in water:MeCN).

General procedure for alkylation of an in situ formed iodine-containing compound.

The oven-dried vessel was equipped with a magnetic stirrer armature coated with PTFE, and 2 eq. PPh ₃ and 2 eq. imidazole, and then DCM (5 ml/mmol) was added. To the resulting mixture was added in portions 2 eq. iodine, then stirred for 15 minutes at room temperature. To the resulting mixture was added 1 eq. suitable alcohol dissolved in DCM and stirred at room temperature. until the transformation ceases to occur. 5 equiv. was added to the formed iodine-containing compound. suitable amine, and the mixture was then stirred for 30 minutes at RT, during which time the reaction had reached a state of complete conversion. Celite was added to the reaction mixture, and volatiles were removed under reduced pressure. The crude product was then purified by flash chromatography using DCM and MeOH (1.2% NH3) as eluents.

General procedure for the alkylation of silyl-protected phenols.

Mixture 1 eq. silyl-protected phenol, 1 eq. alkyl iodide and 1.15 eq. TBAF (1 M in THF) in THF (2 ml/mmol) was stirred at RT. Once a suitable conversion was achieved, volatiles were removed under reduced pressure and the crude intermediate was purified using flash column chromatography.

General procedure for deprotection and hydrolysis.

Mixture 1 eq. substrate and 100 eq. HFxPyr in MeCN (15 ml/mmol) was stirred at 60°C. After reaching a suitable degree of conversion, volatiles were removed under reduced pressure and the residue was suspended in a 1:1 mixture of 1,4-dioxane - water (30 ml/mmol), treated with 150 eq. LiOHxH ₂ O, and stirred at 60°C. After reaching a suitable degree of conversion, volatiles were removed under reduced pressure and the crude product was purified by flash column chromatography using DCM and MeOH (containing 1.2% NH3) as eluents.

General procedure for the formation of a quaternary salt.

The oven-dried vessel was equipped with a PTFE-coated magnetic stirrer armature, and 1 eq. tosylate and 20 eq. suitable amine suspended in CyOH (5 ml/mmol). The reaction mixture was then heated to 140°C and stirred at this temperature until no further transformation was observed. The reaction mixture was diluted with DCM and then applied to a DCM-preconditioned silica gel column. The crude product was then purified by flash column chromatography using DCM and MeOH (1.2% NH3) as eluents.

General procedure for deprotecting a quaternary salt.

To a solution of the appropriate quaternary salt in THF (5 ml/mmol) was added 3 eq. TBAF, and then the mixture was stirred at RT. until the transformation ceases to occur. The reaction mixture was evaporated to dryness under reduced pressure. To the suspension 1 eq. desilylated quaternary salt in dry MeCN (15 ml/mmol) was added 100 equiv. HFxPyr, and then the mixture was stirred at 60°C. After reaching a suitable degree of conversion, the volatiles were removed under reduced pressure, the residue was suspended in a 1:1 mixture of THF - water (30 ml/mmol), 150 equiv was added. LiOHxH ₂ O, and the mixture was stirred at room temperature. After reaching a suitable degree of conversion, the volatiles were removed under reduced pressure. The crude product was purified by flash column chromatography using DCM and MeOH (containing 1.2% NH3) as eluents.

General procedure for alkylation, deprotection and hydrolysis.

A mixture of tertiary amine (1 eq.) and alkalizing agent (10 eq.) in acetonitrile (3 ml/mmol) was stirred at room temperature. Once a suitable degree of conversion was achieved, volatiles were removed under reduced pressure and the crude product was purified by reverse phase flash column chromatography if necessary, or the residue was directly dissolved in acetonitrile (3 ml/mmol), HFxPyr (100 eq.) was added. ), and the mixture was stirred at 60°C. After reaching a suitable degree of conversion, the volatiles were removed under reduced pressure, the residue was suspended in a 1:1 mixture of 1,4-dioxane - water (10 ml/mmol), LiOHxH2O (150 eq.) was added and the mixture was stirred at 60°C. Once a suitable degree of conversion to the desired product was achieved, volatiles were removed under reduced pressure and the crude product was purified using reverse phase flash column chromatography.

General procedure for removing tert-butyldimethylsilyl protecting group.

The appropriate TBS-protected compound and [(1S,4R)-7,7-dimethyl-2-oxonorborne-1yl]methanesulfonic acid (usually 0.25 eq., 1.25 eq. if N,Ndimethylamino moiety is present in the molecule) were mixed in a mixture methanol (2.5 ml/mmol) and dichloromethane (2.5 ml/mmol) in a sealed tube, and stirred at 50°C until complete conversion was observed (typically 1-5 h). After the reaction time, the mixture was poured into a separatory funnel, diluted with DCM and

- 25 046336 washed with conc. NaHCO ₃ solution and water. Dried over MgSO ₄ , filtered through celite and then purified using flash column chromatography.

General procedure for deprotection and hydrolysis of phosphonic acid derivatives.

A solution of the diethylphosphonate derivative in DCM (5 ml/mmol) and MeCN (5 ml/mmol) was purged with nitrogen, then 40 equiv was added. TEA, and the mixture was then cooled to 0°C. 20 eq. was added in one portion. bromo(trimethyl)silane, and then the mixture was stirred for another 30 minutes at 0°C. The mixture was left to warm to room temperature. and stirred until complete conversion to ethyl ester is achieved (typically 1-5 hours). The oc-protecting group(s) were usually cleaved off, but only partially, resulting in a mixture of products. After complete conversion, the mixture was quenched at 0°C with a few mL of MeOH, concentrated on Celite, and then purified by flash column chromatography using EtOAc and MeOH (containing 1.2% NH3) as eluents. The two products (with Boc, without Boc) were combined and proceeded to the General Deprotection and Hydrolysis Procedure.

General procedure for amine substitution and hydrolysis reactions.

To the product of Synthesis 14 in a 1:1 mixture of acetonitrile and Na-methyl-2-pyrrolidone (10 ml/mmol), a suitable amine (3-10 eq.) was added, and the mixture was stirred at 50°C for 2-24 hours. After adding 70% HF in pyridine (50-100 eq.) at RT, the mixture was stirred for 4-18 hours. After purification of the substitution product by column chromatography (silica gel, using DCM and MeOH as eluents), the product was dissolved in THF (8 ml/mmol), and water (2 ml/mmol) and LiOHxH ₂ O (5 eq.) were added and the mixture was stirred at 20-40°C for 1-4 hours. The hydrolyzed product was purified using preparative HPLC (using acetonitrile and 5 µM aqueous NH4HCO3 as eluents) to obtain the desired product.

Syntheses

The following experimental details describe the preparation of synthetic intermediates.

Synthesis 1a: methyl 2-{[(tert-butoxy)carbonyl]amino}-5-[3-(2-fluoro-4-iodophenoxy)propyl]-1,3thiazol-4-carboxylate.

Step A: Methyl 2-(tert-butoxycarbonylamino)-5-iodothiazole-4-carboxylate.

50.00 g of methyl 2-(tert-butoxycarbonylamino)thiazole-4-carboxylate (193.55 mmol, 1 eq.) was suspended in 600 ml of dry MeCN. 52.25 g of N-iodosuccinimide (232.30 mmol, 1.2 eq.) was added and the resulting mixture was stirred overnight at room temperature. The reaction mixture was diluted with brine and then extracted with EtOAc. The combined organic layers were extracted with 1 M Na ₂ S ₂ O ₃ solution, then with brine again. Then it was dried over Na ₂ SO ₄ , filtered and the filtrate was concentrated under reduced pressure. The crude product was purified by flash column chromatography using heptane as eluent to give 60 g (156 mmol, 80%) of the expected product.

1H NMR (400 MHz, DMSO δ ppm 12.03/11.06 (br s), 3.78 (s, 3H), 1.47 (s, 9H); ¹³ C NMR (400 MHz, DMSO-de) δ ppm 153.8, 82.5, 77.7, 52.3, 28.3; MSHR-ERI (m/z): [M+H]+, calculated for C ₁₀ H ₁₄ IN ₂ O ₄ S: 384.9713; found: 384.9708.

Step B: Methyl 2-(tert-butoxycarbonylamino)-5-(3-hydroxyprop-1-ynyl)thiazole-4-carboxylate.

A 500 mL oven-dried, single-neck, round-bottom flask was equipped with a PTFE-coated magnetic stir bar and a reflux condenser was installed. It was loaded with 9.6 g of the product from step A (25 mmol, 1 eq.), 2.80 g of prop-2-yn-1-ol (2.91 ml, 50 mmol, 2 eq.) and 36.10 g of DIPA (50 ml, 356.8 mmol , 14.27 eq.), then 125 ml of dry THF was added, and the system was purged with argon. After 5 min of stirring in an inert atmosphere, 549 mg of Pd(PPh ₃ ) ₂ Cl ₂ (1.25 mmol, 0.05 eq.) and 238 mg of CuI (1.25 mmol, 0.05 eq.) were added. The resulting mixture was then heated to 60°C and stirred at this temperature until no further transformation was observed. Celite was added to the reaction mixture, and volatiles were removed under reduced pressure. The crude product was then purified by flash column chromatography using heptane and EtOAc as eluents to obtain 7.30 g (23 mmol, 93%) of the title product as a yellow solid.

1H NMR (400 MHz, DMSOO δ ppm 12.10 (br s, 1H), 5.45 (t, 1H), 4.36 (d, 2H), 3.79 (s, 3H), 1.48 (s, 9H); ¹³ C NMR (125 MHz, DMSO^ ₆ ) δ ppm 161.3, 142.4, 118.1, 101.4, 73.9, 52.4, 50.2, 28.3; HRMS-ERI (m/z): [M+H]+, calculated for C ₁₃ H ₁₇ N ₂ O ₅ S: 313.0853; found: 313.0866.

Step C: Methyl 2-(tert-butoxycarbonylamino)-5-(3-hydroxypropyl)thiazole-4-carboxylate.

A 1 L oven-dried pressure reaction vessel equipped with a PTFE-coated magnetic stir bar was charged with 44.75 g of the product from step B (143.3 mmol, 1 eq.) and 7.62 g of Pd/C (7.17 mmol, 0.05 eq. ) in 340 ml ethanol, and then created a nitrogen atmosphere using a hydrogenation system. The vessel was then filled with H ₂ gas at a pressure of 4 bar and the reaction mixture was stirred at room temperature. during the night. Complete conversion was observed, but only the olefin product was formed. After filtering the catalyst through a celite pad, the entire procedure was repeated with 5 mol.% new catalyst. The resulting mixture was stirred overnight to ensure complete conversion. Celite was added to the reaction mixture, and volatile substances

- 26 046336 was removed under reduced pressure. The crude product was then purified by flash column chromatography using heptane and EtOAc as eluents to obtain 31.9 g (101 mmol, 70%) of the title product as light yellow crystals.

¹ H NMR (500 MHz, DMSO-de) δ ppm 11.61 (br s, 1H), 4.54 (t, 1H), 3.76 (s, 3H), 3.43 (m, 2H), 3.09 (t, 2H), 1.74 (m, 2H), 1.46 (s, 9H); ¹³ C NMR (125 MHz, DMSO-de) δ ppm 162.8, 143.1, 135.4, 60.3, 51.9, 34.5, 28.3, 23.4; MSHR-ERI (m/z): [M+H]+, calculated for C ₁₃ H ₂₁ N ₂ O ₅ S: 317.1166; found: 317.1164.

Step D: Methyl 2-{[(tert-butoxy)carbonyl]amino}-5-[3-(2-fluoro-4-iodophenoxy)propyl]-1,3thiazol-4-carboxylate.

A 250 mL oven-dried, single-neck, round-bottomed flask equipped with a PTFE-coated magnetic stir bar was charged with 3.40 g of 2-fluoro-4-iodophenol (14 mmol, 1 eq.), 5.00 g of the product from step C (16 mmol , 1.1 eq.) and 4.10 g of PPh ₃ (16 mmol, 1.1 eq.), and 71 ml of dry toluene. After stirring for 5 min under nitrogen, 3.10 mL of DIAD (3.20 g, 16 mmol, 1.1 eq.) was added in one portion while heating the reaction mixture. The reaction mixture was then heated to 50°C and stirred at this temperature for 30 minutes, during which time the reaction reached a state of complete conversion. The reaction mixture was directly injected onto a preconditioned silica gel column, and the crude product was then purified by flash column chromatography using heptane and EtOAc as eluents. The crude product was crystallized from MeOH to give 4.64 g (9.24 mmol, 66%) of the title product.

1H NMR (500 MHz, DMSOHO δ ppm 11.64 (br s, 1H), 7.59 (dd, 1H), 7.45 (dd, 1H), 6.98 (t, 1H), 4.06 (t, 2H), 3.73 ( s, 3H), 3.22 (t, 2H), 2.06 (m, 2H), 1.46 (s, 9H); ¹³ C NMR (125 MHz, DMSO<) δ ppm 134.0, 124.9, 117.6, 68.2, 51.9 , 30.5, 28.3, 23.2; MSHR-ERI (m/z): [M+H]+, calculated for C ₁₉ H ₂₃ N ₂ O ₅ FSI: 537.0351; found: 537.0348.

Synthesis 1b: Methyl 2-(tert-butoxycarbonylamino)-5-[3-[4-[3-tert-butoxycarbonyl(methyl)amino]prop-1-ynyl]-2-fluorophenoxy]propyl]thiazole-4-carboxylate.

A 500 mL oven-dried, single-neck, round-bottom flask was equipped with a PTFE-coated magnetic stir bar and a reflux condenser was installed. It was loaded with 13.41 g of synthesis product 1a (25 mmol, 1 eq.), 8.46 g of tert-butyl N-methyl-N-prop-2-ynylcarbamate (50 mmol, 2 eq.) and 50 ml of DIPA (36.10 g, 50 ml , 356.8 mmol, 14.27 eq.), then 125 ml of dry THF was added and the system was purged with argon. After 5 min of stirring in an inert atmosphere, 549 mg of Pd(PPh ₃ ) ₂ Cl ₂ (1.25 mmol, 0.05 eq.) and 238 mg of CuI (1.25 mmol, 0.05 eq.) were added. The resulting mixture was then heated to 60°C and stirred at this temperature until no further transformation was observed. Celite was added to the reaction mixture, and volatiles were removed under reduced pressure. The crude product was then purified by flash column chromatography using heptane and EtOAc as eluents to obtain 10.5 g (18.2 mmol, 73%) of the desired product.

1H NMR (500 MHz, DMSO<) δ ppm 11.65 (br s, 1H), 7.31 (br d, 1H), 7.21 (br d, 1H), 7.14 (t, 1H), 4.23 (s, 2H), 4.10 (t, 2H), 3.73 (s, 3H ), 3.23 (t, 2H), 2.86 (s, 3H), 2.07 (m, 2H), 1.46/1.41 (s, 18H); ¹³ C NMR (125 MHz, DMSO-de) δ ppm 129.1, 119.2, 115.4, 68.1, 51.9, 38.6, 33.8, 30.5, 23.2; MSHR-ERI (m/z): [M+H] ⁺ , calculated for C ₂₈ H ₃₇ FN ₃ O ₇ S: 578.2331; found: 578.2331.

Synthesis 1c: methyl 2-(tert-butoxycarbonylamino)-5-[3-[4-[3-(dimethylamino)prop-1-ynyl]-2fluorophenoxy]propyl]thiazole-4-carboxylate.

A 250 mL oven-dried, single-neck, round-bottom flask was equipped with a PTFE-coated magnetic stir bar and a reflux condenser was installed. It was loaded with 5.36 g of synthesis product 1a (10 mmol, 1 eq.), 1.66 g of KH-dimethylprop-2-yn-1-amine (20 mmol, 2 eq.) and 20 ml of DIPA (142.7 mmol, 14.27 eq.) , then 50 ml of dry THF was added and the system was purged with argon. After 5 min of stirring in an inert atmosphere, 220 mg of Pd(PPh ₃ ) ₂ Cl ₂ (0.5 mmol, 0.05 eq.) and 95 mg of CuI (0.5 mmol, 0.05 eq.) were added. The resulting mixture was then heated to 60°C and stirred at this temperature until no further transformation was observed. Celite was added to the reaction mixture, and volatiles were removed under reduced pressure. The crude product was then purified by flash column chromatography using DCM and MeOH (1.2% NH ₃ ) as eluents to give 4.5 g (7.8 mmol, 78%) of the title product.

1H NMR (500 MHz, DMSOO δ ppm 11.66 (s, 1H), 7.29 (dd, 1H), 7.19 (m, 1H), 7.12 (t, 1H), 4.09 (t, 2H), 3.73 (s , 3H), 3.44 (s, 2H), 3.23 (t, 2H), 2.24 (s, 6H), 2.07 (m, 2H), 1.45 (s, 9H); ¹³ C NMR (125 MHz, DMSO<) δ ppm 162.8, 147.3, 129.0, 119.2, 115.4, 84.3, 68.0, 51.9, 48.1, 44.2, 30.6, 28.3, 23.2; HRMS-ERI (m/z): [M+H]+, calculated for C ₂₄ H ₃₁ FN ₃ O ₅ S: 492.1963; found: 492.1956.

Synthesis 2a: 5-tert-Butyl(dimethyl)silyl]oxy-4-tert-butyl(diphenyl)silyl]oxypentan-1-ol.

Step A: pent-4-enyl benzoate.

30.00 g of pent-4-en-1-ol (0.35 mol, 1 eq.) and 58.5 ml of N.N-diethylethanamine (0.42 mol, 1.2 eq.) were mixed in 200 ml of DCM, then cooled to 0°C. 48.5 mL of benzoyl chloride (0.42 mol, 1.2 eq.) was added to the mixture at 0°C using a dropping funnel under an inert atmosphere. After addition, the mixture was further stirred at 0°C for 30 min, then at room temperature. VN. The mixture was diluted with 100 ml DCM, then the organic phase was washed with water, 1 M NaOH, 1 M HCl and brine, respectively.

- 27 046336

The organic phase was dried over MgSO ₄ , filtered, concentrated and purified by flash column chromatography using heptane and EtOAc as eluents to obtain 63.19 g (95%) of the title product as a colorless liquid.

1H NMR (500 MHz, DMSOHO δ ppm 7.97 (dd, 2H), 7.66 (t, 1H), 7.53 (t, 2H), 5.91-5.81 (m, 1H), 5.094.97 (m, 2H) , 4.27 (t, 2H), 2.17 (q, 2H), 1.81 (qv, 2H); ¹³ C NMR (125 MHz, DMSO-de) δ ppm 166.2, 138.2, 133.8, 130.3, 129.6, 129.2, 115.8, 64.5, 30.1, 27.8; GC-MS-EI (m/z): [M] ⁺ , calculated for C ₁₂ H ₁₄ O ₂ : 190.1; found: 190.

Step B: 4,5-dihydroxypentyl benzoate.

42.22 g of the product from step A (0.26 mol, 1.0 eq.) and 50.40 g of 4-methyl-4-oxidomorpholin-4-ium hydrate (0.37 mol, 1.7 eq.) were mixed in 360 ml of 2-methylpropan-2-ol and 40 ml of water, then 6.57 g of osmium tetroxide (2.5 wt.% in 2-methylpropan-2-ol, 0.64 mmol, 0.002 eq.) was added and the mixture was stirred at 60°C for 24 hours. Complete conversion was observed. The mixture was cooled to room temperature. and added 1 M Na ₂ S ₂ O ₃ , then stirred for another 10 minutes at room temperature. DCM was added and the organic phase was separated, washed with water and brine, respectively. The solution was dried over MgSO ₄ , filtered, concentrated and purified by flash column chromatography using heptane and EtOAc as eluents to obtain 36.9 g (63%) of the title product as a white solid.

1H NMR (500 MHz, DMSOO δ ppm 7.99-7.50 (m, 5H), 4.50 (m, 2H), 4.28 (m, 2H), 3.45 (m, 1H), 3.303.24 (m+m, 2H), 1.85-1.72 (m+m, 2H), 1.59-1.33 (m+m, 2H); ¹³ C NMR (125 MHz, DMSO<) δ ppm 166.2, 133.8-129.1, 71.2, 66.3, 65.5, 30.3, 25.2; MSHR-ERI (m/z): [M+Na] ⁺ , calculated for CuHusNaO^ 247.0941; found: 247.0941.

Step C: 5-[tert-butyl(dimethyl)silyl]oxy-4-hydroxypentyl]benzoate.

24.86 g of the product from step B (0.11 mol, 1 eq.) and 15.09 g of imidazole (0.22 mol, 2 eq.) were mixed in 120 ml of NN-dimethylformamide and then cooled to -20°C in an inert atmosphere. 16.71 g of tert-butylchlorodimethylsilane (0.11 mol, 1 eq.) in 40 ml of N,Ndimethylformamide were added slowly over a period of 30 min, supplemented with 10 ml of DCM, then allowed to warm to room temperature. and additionally stirred the VN. We observed a complete transformation. Extinguished the conc. NH4C1 solution, then most of the volatile substances were evaporated. EtOAc and water were added to the residue, the organic phase was separated, then washed with water and brine, dried over MgSO ₄ , filtered, concentrated and purified by flash column chromatography using heptane and EtOAc as eluents to obtain 33.71 g (90%) the target product in the form of a colorless oil.

1H NMR (500 MHz, DMSO<) δ ppm 7.95 (m, 2H), 7.66 (m, 1H), 7.52 (m, 2H), 4.58 (d, 1H), 4.29 (m, 2H), 3.51-3.35 (dd+dd, 2H), 3.48 (m, 1H), 1.86-1.74 (m+m, 2H), 1.67-1.34 (m+m, 2H), 0.83 (s, 9H), 0.01 (s, 6H); ¹³ C NMR (125 MHz, DMSO<) δ ppm 166.2, 133.7, 130.4, 129.5, 129.2, 70.6, 67.7, 65.3, 30.2, 26.3, 24.9, -4.9.

Step D: [5-[tert-butyl(dimethyl)silyl]oxy-4-[tert-butyl(diphenyl)silyl]oxypentyl]benzoate.

33.51 g of the product from step C (0.10 mol, 1 eq.), 16.85 g of imidazole (0.25 mol, 2.5 eq.) and 1.21 g of N,Ndimethylpyridin-4-amine (0.01, 0.1 eq.) were mixed in 230 ml of N,N -dimethylformamide, then slowly added 38 ml of tert-butylchlorodiphenylsilane (0.15 mol, 1.5 eq.), supplemented with 20 ml of N,Ndimethylformamide and then stirred at 50°C overnight. We observed a complete transformation. The mixture was cooled to room temperature, quenched with conc. NH ₄ Cl solution, then most of the volatile substances were evaporated. EtOAc and water were added to the residue, the organic phase was separated, then washed with water and brine, dried over MgSO4, filtered, concentrated and purified by flash column chromatography using heptane and EtOAc as eluents to obtain 56.43 g (99%) of the desired product in the form of a colorless viscous oil.

1H NMR (500 MHz, DMSO-de) δ ppm 7.91-7.37 (m, 15H), 4.17 (m, 2H), 3.76 (m, 1H), 3.45 (m, 2H), 1.72 (m, 2H), 1.66-1.57 (m+m, 2H), 0.99 ( s, 9H), 0.74 (s, 9H), -0.12/-0.16 (s+s, 6H); ¹³ C NMR (125 MHz, DMSO^) δ ppm 166.1, 136.0-128.0, 73.3, 66.0, 65.1, 30.3, 27.3, 26.1, 24.0, -5.1; HRMS-ERI (m/z): [M+Na] ⁺ , calculated for C ₃ 4H48NaO4Si2: 599.2983; found: 599.2981.

Step E: 5-[tert-butyl(dimethyl)silyl]hydroxy-4-[tert-butyl(diphenyl)silyl]oxypentan-1-ol.

46.10 g of the product from step D (0.08 mol, 1 eq.) was dissolved in 227 ml MeOH and 117 ml THF, and then 12.79 g NaOH (0.32 mol, 4.0 eq.) in 85 ml water was slowly added while the mixture was cooled with ice. After addition, the mixture was left to stir at room temperature. until complete conversion was observed (approx. 4 hours). EtOAc and water were added, the mixture was separated and the organic phase was washed with brine, dried over MgSO ₄ , filtered, concentrated and purified by flash column chromatography using heptane and EtOAc as eluents to give 29.32 g (78%) of the title product in in the form of a colorless oil.

1H NMR (500 MHz, DMSO<) δ ppm 7.65-7.37 (m, 10H), 4.34 (t, 1H), 3.71 (m, 1H), 3.42 (m, 2H), 3.26 (m, 2H), 1.52 (m, 2H), 1.42 (m, 2H) , 0.99 (s, 9H), 0.77 (s, 9H), -0.13 (s, 6H); ¹³ C NMR (125 MHz, DMSOO δ ppm 135.8, 135.8, 134.3, 134.0, 130.3, 130.2, 128.2, 128.0, 74.0, 66.4, 61.4, 30.4, 28.3, 27.3, 26.2 , -5.1; MSVRERI (m/ z): [M+Na] ⁺ , calculated for C ₂₇ H ₄₄ NaO ₃ Si ₂ : 495.2721; found: 495.2706.

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Synthesis 2b: 5-[tert-butyl(dimethyl)silyl]hydroxy-4-methoxypentan-1-ol.

Step A: [5-[tert-butyl(dimethyl)silyl]oxy-4-methoxypentyl]benzoate.

3.66 g of synthesis product 2a, step C (10.8 mmol, 1 equiv.), 6.95 g of H1,H1,III,III-tetramethylna(|)talin1,8-diamine (32.4 mmol, 3 equiv.) and a small portion of 4A molecular sieves mixed in 210 ml DCM, then 4.00 g trimethyloxonium tetrafluoroborate (27.0 mmol, 2.5 eq.) was added in one portion and the mixture was stirred at room temperature. within 5 hours. Complete transformation was observed. The mixture was filtered through a pad of celite, washed with DCM, then concentrated. The residue was redissolved in DCM and washed with water, 1 M CuSO ₄ and brine, respectively. The solution was dried over MgSO ₄ , filtered, concentrated on celite and purified by flash column chromatography using heptane and EtOAc as eluents to obtain 2.58 g (68%) of the title product as a colorless oil.

Ή NMR (400 MHz, DMSO<) δ ppm 7.95 (d, 2H), 7.65 (t, 1H), 7.51 (t, 2H), 4.27 (t, 2H), 3.56 (ddd, 2H), 3.30 (s, 3H), 3.23-3.18 (m, 1H) , 1.84-1.44 (m, 4H), 0.83 (s, 9H), 0.02 (s, 6H); ¹³ C NMR (100 MHz, DMSO<) δ ppm 165.7, 133.2, 129.9, 129.0, 128.7, 80.5, 64.7, 63.9, 57.0, 27.2, 25.7, 24.2, 17.9, -5.5, -5.5.

Step B: 5-[tert-butyl(dimethyl)silyl]oxy-4-methoxypentan-1-ol.

2.54 g of the product from step A (7.20 mol, 1 eq.) was dissolved in 54 ml of MeOH, then 1.44 g of NaOH (36 mmol, 5.0 eq.) in 18 ml of water was slowly added to the mixture. After addition, the mixture was stirred at room temperature. until complete conversion was observed (approx. 1.5 h). EtOAc and water were added, the mixture was separated and the organic phase was washed with brine, dried over MgSO ₄ , filtered and concentrated to give 1.76 g (98%) of the title product as a light yellow oil.

Ή NMR (400 MHz, DMSO<) δ ppm 4.37 (t, 1H), 3.58-3.50 (m, 2H), 3.39-3.32 (m, 2H), 3.29 (s, 3H), 3.17-3.10 (m, 1H), 1.51-1.32 (m, 4H), 0.86 (s, 9H), 0.03 (s, 6H); ¹³ C NMR (100 MHz, DMSO<) δ ppm 81.0, 64.4, 60.9, 57.0, 28.4, 27.2, 25.8, 18.0, -5.4, -5.4; LC-MS-ESI (m/z): [M+H]+, calculated for C ₁₂ H ₂₉ O ₃ Si: 249.2; found: 249.2.

Synthesis of 2c: 5-methoxy-4-triisopropylsilyloxypentan-1-ol.

Step A: [5-[tert-butyl(dimethyl)silyl]oxy-4-triisopropylsilyloxypentyl]benzoate.

2.0 g of synthesis product 2a, step C (5.91 mmol, 1 eq.), and 1.21 g of imidazole (17.73 mmol, 3 eq.) were mixed in 6 ml of N,N-dimethylformamide, then 1.71 g of chloro(triisopropyl) )silane (8.86 mol, 1.5 eq.), and the mixture was stirred at 60°C for 3.5 hours. Additional amounts of chloro(triisopropyl)silane (0.5 eq.) and imidazole (1 eq.) were added and stirred for another 3 h. We observed a complete transformation. The mixture was cooled to RT, then EtOAc and water were added, the organic phase was separated, washed with brine, dried over MgSO ₄ , filtered, concentrated on Celite and purified by flash column chromatography using heptane and EtOAc as eluents to give 2.27 g (99%) of the target product in the form of a colorless viscous oil.

Ή NMR (400 MHz, DMSO<) δ ppm 7.94 (d, 2H), 7.66 (t, 1H), 7.52 (t, 2H), 4.29 (t, 2H), 3.86 (qv, 1H), 3.53 (ddd, 2H), 1.83-1.56 (m, 4H) , 1.02 (m, 21H), 0.84 (s, 9H), 0.02 (s, 6H).

Step B: (5-hydroxy-4-triisopropylsilyloxypentyl)benzoate.

Using the general tert-butyldimethylsilyl deprotection procedure from 2.25 g of the product from step A (4.54 mmol, 1 eq.) followed by flash column chromatography using heptane and EtOAc as eluents, 1.13 g (65%) of the desired product was obtained product.

Ή NMR (400 MHz, DMSO<) δ ppm 7.95 (d, 2H), 7.65 (t, 1H), 7.52 (t, 2H), 4.63 (t, 1H), 4.29 (qv, 2H), 3.85-3.78 (m, 1H), 3.45-3.26 (m, 2H), 1.83-1.54 (m, 4H), 1.01 (s, 21H); LC-MS-ESI (m/z): [M+H] ⁺ , calculated for C ₂ iH ₃₇ O ₄ Si: 381.2; found: 381.3.

Step C: (5-methoxy-4-triisopropylsilyloxypentyl)benzoate.

1.11 g of the product from step B (2.91 mmol, 1 eq.) and 1.87 g of H1.H1.H8.H8-tetramethylnaphthalene-1.8diamine (8.7 mmol, 3 eq.) were mixed in 45 ml of DCM, then 1.87 was added in one portion g trimethyloxonium tetrafluoroborate (8.7 mmol, 3 eq.), and the mixture was stirred at room temperature. within 4 hours. Complete transformation was observed. The mixture was filtered through a pad of celite, washed with DCM, then the organic phase was washed with water, 1 M CuSO ₄ and brine, respectively. The solution was dried over MgSO ₄ , filtered, concentrated on celite and purified by flash column chromatography using heptane and EtOAc as eluents to obtain 963 mg (84%) of the title product as a colorless oil.

¹ H NMR (400 MHz, DMSO<) δ ppm 7.95 (d, 2H), 7.66 (t, 1H), 7.52 (t, 2H), 4.28 (t, 2H), 3.96 (qv, 1H), 3.31 (d, 2H), 3.25 (s, 3H), 1.82 -1.54 (m, 4H), 1.01 (m, 21H); ¹³ C NMR (100 MHz, DMSOL) δ ppm 165.7, 133.3, 129.8, 129.0, 128.7, 75.9, 70.4, 64.7, 58.4, 30.7, 23.5, 18.0, 17.9, 12.0.

Step D: 5-methoxy-4-triisopropylsilyloxypentan-1-ol.

953 mg of the product from step C (2.41 mmol, 1 eq.) was dissolved in 12 ml MeOH and 6 ml THF, then 487 mg NaOH (12.1 mmol, 5.0 eq.) in 6 ml water was slowly added. After addition, the mixture was left to stir at room temperature. until complete conversion was observed (approx. 2 h). Added

- 29 046336

EtOAc and water, the mixture was then separated and the organic phase was washed with brine, dried over MgSO ₄ , filtered, concentrated on Celite and purified by flash column chromatography using heptane and EtOAc as eluents to give 687 mg (78%) of the title product as colorless oil.

1H NMR (500 MHz, DMSOHO δ ppm 4.39 (t, 1H), 3.89 (m, 1H), 3.37 (m, 2H), 3.27 (d, 2H), 3.24 (s, 3H), 1.47 (m , 2H), 1.47 (m, 2H), 1.03 (m, 21H), ¹³ C NMR (125 MHz, DMSO-de) δ ppm 76.8, 71.4, 61.4, 58.8, 31.4, 28.3, 18.5, 12.5; MSHR-ERI (m/z): [M+H]+, calculated for C ₁₅ H ₃₅ O ₃ Si: 291.2350; found: 291.2349.

Synthesis 2d: 4-[tert-Butyl(dimethyl)silyl]oxy-5-(dimethylamino)pentan-1-ol.

Step A: 3-(oxiran-2-yl)propyl benzoate.

19.01 g of 3-chlorobenzenecarboperoxic acid (77 wt.%, 85.83 mmol, 1.3 eq.) was dissolved in DCM (3 ml/mmol) in a separating funnel, then the organic phase (3-4 ml of water was discarded) was directly added to the solution of 12.41 g of product synthesis 2a, step A (65.25 mmol, 1 eq.), in DCM (3 ml/mmol), and stirred at room temperature. within 4 hours. Complete transformation was observed. Add 100 ml DCM, then wash with 2x100 ml conc. NaHCO ₃ solution and 1 x 100 ml brine, dried over MgSO4, filtered, concentrated and purified by flash column chromatography using heptane and EtOAc as eluents to obtain 13.92 g (quantity) of the title product as a colorless oil.

1H NMR (400 MHz, DMSOD ₆ ) δ ppm 7.97 (dm, 2H), 7.66 (tm, 1H), 7.53 (tm, 2H), 4.31 (m, 2H), 2.97 (m, 1H), 2.68-2.48 (dd+dd, 2H), 1.84 (m, 2H), 1.67-1.56 (m+m, 2H); ¹³ C NMR (100 MHz, DMSOL) δ ppm 166.2, 133.8, 129.6, 129.3, 64.8, 51.7, 46.6, 29.1, 25.4; MSVR-EI (m/z): [M] ⁺ , calculated for C12H14O3: 206.0943; found: 206.0941.

Step B: [4-[tert-butyl(dimethyl)silyl]oxy-5-(dimethylamino)pentyl]benzoate.

7.01 g of the product from step A (34 mmol, 1 eq.) and 51 ml of dimethylamine solution (2 M in MeOH, 102 mmol, 3 eq.) were mixed in a sealed tube and stirred at 60°C for 1 h. Complete conversion was observed . The reaction mixture was concentrated and [5-(dimethylamino)-4-hydroxypentyl]benzoate was obtained as a viscous light yellow oil. MSHR-ERI (m/z): [M+H] ⁺ , calculated for Ci ₄ H ₂₂ NO ₃ : 252.1594; found: 252.1597.

The crude product was redissolved in NN-dimethylformamide (2 ml/mmol), then 6.95 g of imidazole (102 mmol, 3 eq.), 208 mg of X,X-dimethylpyridin-4-amine (1.70 mmol, 0.05 eq.) and, After the mixture cleared, a solution of 12.8 g of tert-butylchlorodimethylsilane (85.0 mmol, 2.5 eq.) was added in one portion. The mixture was stirred at 60°C for 3 hours (>95% conversion observed). The mixture was cooled to room temperature, then 3-4 ml of conc. NH4C1 solution, stirred for 5 minutes, then the volatile substances were evaporated. Added 300 ml EtOAc, 50 ml water and 50 ml conc. NaHCO ₃ solution, and then the organic layer was separated. The organic phase was washed with 1x50 ml H ₂ O, 1x50 ml brine, then dried over MgSO ₄ , filtered and then concentrated. The crude product was purified by flash column chromatography using DCM and MeOH as eluents to obtain 8.37 g (67%) of the title product as a colorless oil.

1H NMR (400 MHz, DMSO<) δ ppm 7.99-7.49 (m, 5H), 4.29-4.27 (m+m, 2H), 3.78 (m, 1H), 2.20-2.17 (dd+dd, 2H), 2.13 (s, 6H), 1.80-1.73 (m +m, 2H), 1.67-1.46 (m+m, 2H), 0.84 (s, 9H), 0.05 (s, 3H), 0.04 (s, 3H); ¹³ C NMR (100 MHz, DMSO<) δ ppm 166.2, 130.3, 70.0, 66.0, 65.3, 46.6, 32.0, 26.3, 24.5, -3.8, -4.4; MSVRERI (m/z): [M+H]+, calculated for C ₂₀ H ₃₆ NO ₃ Si: 366.2459; found: 366.2463.

Step C: 4-[tert-butyl(dimethyl)silyl]oxy-5-(dimethylamino)pentan-1-ol.

3.12 g of the product from step B (8.53 mmol, 1 eq.) was dissolved in 34 ml of MeOH and 17 ml of THF, then 1.36 g of NaOH (34 mmol, 4.0 eq.) in 17 ml of water was slowly added. After addition, the mixture was left to stir at room temperature. until complete conversion was observed (approx. 1.5 h). EtOAc and water were added, the mixture was separated and the organic phase was washed with brine, dried over MgSO4, filtered, concentrated on Celite and purified by flash column chromatography using DCM and MeOH as eluents to give 1.60 g (72%) of the title product as colorless oil.

1H NMR (400 MHz, CDCl ₃ ) 3.83-3.76 (m, 1H), 3.65-3.55 (m, 3H), 2.31 (ddd, 2H), 2.22 (s, 6H), 1.72-1.54 (m, 4H), 0.87 (s, 9H), 0.05 (s, 3H), 0.05 (s, 3H); ¹³ C NMR (100 MHz, CDCl ₃ ) δ ppm; 70.1, 65.2, 62.9, 46.4, 31.9, 27.9, 26.0, 18.2, -4.36, -4.62; MSHR-ERI (m/z): [M+H]+, calculated for C ₁₃ H ₃₂ NO ₂ Si: 262.2197; found: 262.2197.

Synthesis 2e: 4-[tert-Butyl(dimethyl)silyl]oxy-5-morpholino-pentan-1-ol.

Step A: [4-[tert-butyl(dimethyl)silyl]oxy-5-morpholinopentyl]benzoate.

1.50 g of the product of synthesis 2d, step A (7.27 mmol, 1 eq.), and 1.91 g of morpholine (21.87 mmol, 3 eq.) were stirred in 15 ml of MeCN at 82°C for 24 h. The reaction mixture was concentrated and (4hydroxy- 5-morpholinopentyl)benzoate was obtained as a viscous light yellow oil. LC-MS-ESI (m/z): [M+H] ⁺ , calculated for Q6H24NO4: 294.2; found: 294.2.

The crude product was redissolved in 7 ml of N,N-dimethylformamide, then 1.19 g of imidazole (17.52 mmol, 3 eq.) was added and, after the mixture cleared, a solution of 1.76 g was added in one portion

- 30 046336 tert-butylchlorodimethylsilane (11.68 mmol, 2 eq.). The mixture was stirred at 60°C for 2 hours. The mixture was cooled to RT, then EtOAc was added, and the organic layer was washed with H ₂ O, brine, then dried over MgSO ₄ , filtered and then concentrated on celite. The crude product was purified by flash column chromatography using heptane and EtOAc as eluents to obtain 2.18 g (92%) of the title product as a colorless oil.

1H NMR (400 MHz, DMSO-de) δ ppm 7.96 (dd, 2H), 7.66 (t, 1H), 7.53 (t, 2H), 4.29 (t, 2H), 3.85 (m, 1H), 3.52 (t, 4H), 2.42-2.30 (m, 4H) , 2.26 (t, 2H), 1.85-1.44 (m, 4H), 0.85 (s, 9H), 0.07 (s, 3H), 0.04 (s, 3H); ¹³ C NMR (100 MHz, CDCl ₃ ) δ ppm 165.7, 133.3, 129.9, 129.0, 128.7, 68.7, 66.2, 64.8, 64.7, 54.2, 31.6, 25.8, 24.1, 17.8, -4.2, -4.8; LC-MS-ESI (m/z): [M+H]+, calculated for C ₂₂ H ₃₈ NO ₄ Si: 408.3; found: 408.3.

Step B: 4-[tert-butyl(dimethyl)silyl]oxy-5-morpholinopentan-1-ol.

2.18 g of the product from step A (8.53 mmol, 1 eq.) was dissolved in 20 ml of MeOH and 20 ml of THF, then 2.22 g of K2C.O ₃ (16.06 mmol, 3.0 eq.) was added in one portion. After addition, the mixture was left to stir at room temperature. until complete transformation was observed. Most of the volatiles were evaporated, then EtOAc and water were added, the mixture was separated and the organic phase was washed with brine, dried over MgSO ₄ , filtered and concentrated. 1.49 g (72%) of the target product was obtained in the form of a colorless oil.

1H NMR (400 MHz, DMSO^ ₆ ) δ ppm 7.97 (d, 2H), 7.66 (t, 1H), 7.53 (t, 2H), 4.37 (t, 1H), 3.80-3.72 (m, 1H), 3.55 (t, 4H), 3.37 (q, 2H) , 2.44-2.29 (m, 4H), 2.22 (ddd, 2H), 1.56-1.29 (m, 4H), 0.85 (s, 9H), 0.06 (s, 3H), 0.04 (s, 3H); ¹³ C NMR (100 MHz, DMSO-de) δ ppm 69.3, 66.2, 65.3, 61.0, 54.3, 32.1, 28.5, 25.8, 17.9, -4.2, -4.7; LC-MS-ESI (m/z): [M+H]+, calculated for Ci5H ₃₄ NO ₃ Si: 304.2; found: 304.3.

Synthesis of 2f: 4-[tert-Butyl(dimethyl)silyl]hydroxy-5-[2-(dimethylamino)ethylmethylamino]pentan-1-ol.

Step A: [4-[tert-butyl(dimethyl)silyl]oxy-5-[2-(dimethylamino)ethylmethylamino]pentyl] benzoate.

908 mg of synthesis product 2d, step A (4.40 mmol, 1 equiv.), and 1.35 g of K.I-trimethylethane-1.2diamine (13.2 mmol, 3 equiv.) were stirred in 12 ml of MeCN at 82°C BH. The reaction mixture was concentrated and [5-[2-(dimethylamino)ethylmethylamino]-4-hydroxypentyl]benzoate was obtained as a viscous light yellow oil. LC-MS-ESI (m/z): [M+H]+, calculated for C ₁₇ H ₂₉ N ₂ O ₃ : 309.2; found: 309.3.

The crude product was redissolved in 4 ml of N,N-dimethylformamide, then 609 mg of imidazole (8.95 mmol, 3 eq.) was added and, after the mixture had cleared, a solution of 898 mg of tert-butylchlorodimethylsilane (5.96 mmol, 2 eq.) was added in one portion. ). The mixture was stirred at 60°C for 2 hours. The mixture was cooled to RT, then EtOAc was added, and the organic layer was washed with H ₂ O, brine, then dried over MgSO ₄ , filtered and then concentrated on celite. The crude product was purified by flash column chromatography using DCM and MeOH as eluents to obtain 1.11 g (88%) of the title product as a colorless oil.

1H NMR (400 MHz, DMSO^) δ ppm 7.95 (d, 2H), 7.66 (t, 1H), 7.52 (t, 2H), 4.28 (t, 2H), 3.81-3.74 (m, 1H), 2.42-2.36 (m, 2H), 2.32-2.26 ( m, 4H), 2.16 (s, 3H), 2.10 (s, 6H), 1.83-1.40 (m, 4H), 0.84 (s, 9H), 0.05 (s, 3H), 0.04 (s, 3H); ¹³ C NMR (100 MHz, DMSOO δ ppm 165.7, 133.3, 129.9, 129.0, 128.7, 69.8, 64.8, 63.9, 57.0, 56.1, 45.4, 43.4, 31.5, 25.8, 24.0, 1 7.8, -4.3, -4.8 ; LC-MS-ESI (m/z): [M+H] ⁺ , calculated for C ₂₃ H ₄₃ N ₂ O ₃ Si: 423.3; found: 423.3.

Step B: 4-[tert-butyl(dimethyl)silyl]oxy-5-[2-(dimethylamino)ethylmethylamino]pentan-1-ol.

3.42 g of the product from step A (8.09 mmol, 1 eq.) was dissolved in 40 ml of MeOH and 20 ml of THF, then 1.62 g of NaOH (40.5 mmol, 5.0 eq.) in 20 ml of water was added in one portion. After addition, the mixture was stirred at room temperature. until complete transformation was observed. EtOAc and water were added, the mixture was separated and the organic phase was washed with brine, dried over MgSO4, filtered and concentrated. 2.60 g (quantitative) of the target product was obtained in the form of a light yellow oil.

1H NMR (500 MHz, DMSO<) δ ppm 3.69 (m, 1H), 3.36 (t, 2H), 2.39 (m, 2H), 2.30 (m, 2H), 2.25 (m, 2H), 2.16 (s, 3H), 2.13 (s, 6H), 1.56 -1.30 (m+m, 2H), 1.50-1.40 (m+m, 2H), 0.85 (s, 9H), 0.04 (s, 3H), 0.03 (s, 3H); ¹³ C NMR (125 MHz, DMSO^) δ ppm 70.7, 64.8, 61.5, 57.5, 56.7, 46.0, 43.8, 32.3, 29.0, 26.3, -3.8, -4.3; HRMS-ERI (m/z): [M+H] ⁺ , calculated for C ₁₆ H ₃₉ N ₂ O ₂ Si: 319.2775; found: 319.2788.

Synthesis of 2g: 3-[tert-butyl(dimethyl)silyl]oxy-4-morpholinobutan-1-ol.

Step A: 2-(oxiran-2-yl)ethyl benzoate.

4.063 g of but-3-enyl benzoate (23.06 mmol, 1 eq.) was dissolved in 50 ml of CHCl ₃ , then 4.97 g of 3-chlorobenzenecarboperoxic acid (77 wt.%, 28.80 mmol, 1.25 eq.) was added and stirred at room temperature. T. VN. 60 ml DCM was added, then washed with conc. NaHCO ₃ solution and brine, dried over MgSO ₄ , filtered, concentrated and purified by flash column chromatography using heptane and EtOAc as eluents to obtain 3.81 g (86%) of the title product as a colorless oil.

1H NMR (400 MHz, DMSO^ ₆ ) δ ppm 7.98 (d, 2H), 7.66 (t, 1H), 7.53 (t, 2H), 4.39 (t, 2H), 3.11-3.05 (m, 1H), 2.73 (t, 1H), 2.53 (dd, 1H) , 2.03-1.82 (m, 2H); ¹³ C NMR (100 MHz, DMSO^6) δ ppm 165.7, 133.4, 129.7, 129.2, 128.8, 62.0, 49.2, 45.9, 31.4; LC-MS-ESI (m/z): [M+H] ⁺ , calculated for CnH1 ₃ O ₃ : 193.1; found: 193.1.

- 31 046336

Step B: [3-[tert-butyl(dimethyl)silyl]hydroxy-4-morpholinobutyl]benzoate.

723 mg of the product from step A (3.76 mmol, 1 eq.) and 983 mg of morpholine (11.28 mmol, 3 eq.) were stirred in 10 ml MeCN at 82°C for 24 h. The reaction mixture was concentrated and (3-hydroxy-4morpholinobutyl )benzoate was obtained in the form of a viscous light yellow oil. LC-MS-ESI (m/z): [M+H]+, calculated for Ci5H ₂₂ NO ₄ : 280.2; found: 280.2.

The crude product was redissolved in 5 ml of CN-dimethylformamide. then 487 mg of imidazole (7.15 mmol, 3 eq.) was added and, after the mixture cleared, a solution of 719 mg of tert-butylchlorodimethylsilane (4.77 mmol, 2 eq.) was added in one portion. The mixture was stirred at 60°C for 2 hours. Cooled to rt, then EtOAc was added, and the organic layer was washed with H2O, brine, then dried over MgSO ₄ , filtered and then concentrated on celite. The crude product was purified by flash column chromatography using heptane and EtOAc as eluents to obtain 669 mg (71%) of the title product as a colorless oil.

1H NMR (400 MHz, DMSOHO δ ppm 7.96 (d, 2H), 7.66 (t, 1H), 7.53 (t, 2H), 4.42-4.26 (m, 2H), 4.04-3.97 (m, 1H) , 3.54 (t, 4H), 2.45-2.27 (m, 6H), 2.08-1.98 (m, 1H), 1.82-1.73 (m, 1H), 0.85 (s, 9H), 0.07 (s, 3H), 0.02 (s, 3H); ¹³ C NMR (100 MHz, DMSOO δ ppm 165.7, 133.3, 129.8, 129.0, 128.8, 66.3, 66.2, 65.1, 61.7, 54.2, 34.3, 25.8, 17.8, -4.2, -5 .1 ; LC-MS-ESI (m/z): [M+H] ⁺ , calculated for C21H ₃ 6NO4Si: 394.2; found: 394.3.

Step C: 3-[tert-butyl(dimethyl)silyl]oxy-4-morpholinobutan-1-ol.

366 mg of the product from step B (0.93 mmol, 1 eq.) was dissolved in 2 ml MeOH and 6.75 ml THF, then 186 mg NaOH (4.65 mmol, 5.0 eq.) in 2.25 ml water was added in one portion. After addition, the mixture was left to stir at room temperature. until complete transformation was observed. EtOAc and water were added, the mixture was separated and the organic phase was washed with brine, dried over MgSO ₄ , filtered and concentrated. 262 mg (97%) of the target product was obtained in the form of a colorless oil.

1H NMR (400 MHz, DMSO-06) δ ppm 4.42 (br., 1H), 3.93-3.87 (m, 1H), 3.55 (t, 4H), 3.52-3.41 (m, 2H), 2.37 (dm, 4H), 2.29-2.20 (m, 2H), 1.73 -1.66 (m, 1H), 1.52-1.44 (m, 1H), 0.85 (s, 9H), 0.06 (s, 3H), 0.04 (s, 3H); ¹³ C NMR (100 MHz, DMSOO δ ppm 66.2, 65.4, 57.4, 54.2, 39.0, 25.8, 17.8, -4.3, -4.8; LC-MS-ESI (m/z): [M+H]+ , calculated for C ₁₄ H ₃₂ NO ₃ Si: 290.2; found: 290.3.

Synthesis 2h: 3-[tert-butyl(dimethyl)silyl]oxy-4-methoxybutan-1-ol.

Step A: Methyl 3-hydroxy-4-methoxybutanoate.

To 1.00 g of methyl 4-methoxy-3-oxobutanoate (6.84 mmol, 1 eq.) in 30 ml of MeOH at 0°C, 264 mg of NaBH ₄ (6.98 mmol, 1 eq.) was added in portions, and the mixture was stirred at 0°C in for 10 hours. After concentration, the residue was diluted with water and extracted with EtOAc. The combined organic phases were dried and concentrated to obtain 0.72 g (71%) of the desired product.

1H NMR (400 MHz, DMSO-d6) δ ppm 4.97 (d, 1H), 3.97 (m, 1H), 3.58 (s, 3H), 3.27 (dd, 1H), 3.24 (s, 3H), 3.20 (dd, 1H), 2.46 (dd, 1H), 2.27 (dd, 1H).

Step B: Methyl 3-[tert-butyl(dimethyl)silyl]oxy-4-methoxybutanoate.

To 8.89 g of the product from step A (60 mmol, 1 eq.) and 4.3 g of imidazole (63.2 mmol, 1.05 eq.) in 100 ml of DMF, 10 g of tert-butylchlorodimethylsilane (66.3 mmol, 1.1 eq.) was added in one portion. and the mixture was stirred for 18 hours. After diluting the mixture with 300 ml brine and extracting with EtOAc, the combined organic phases were dried, concentrated and purified by flash column chromatography using heptane and EtOAc as eluents to obtain 11.77 g ( 75%) of the target product.

1H NMR (400 MHz, DMSOO δ ppm 4.17 (sx, 1H), 3.58 (s, 3H), 3.27 (m. 2H), 3.26 (s, 3H), 2.54 (dd, 1H), 2.32 (dd , 1H), 0.82 (s, 9H), 0.03 (d, 6H).

Step C: 3-[tert-butyl(dimethyl)silyl]oxy-4-methoxybutan-1-ol.

To 10.77 g of the product from step B (41 mmol, 1 eq.) in 288 ml DCM at 0°C, 124 ml DIBAL-H (1 M in THF, 124 mmol, 3 eq.) was slowly added. Then the mixture was stirred for 2.5 h at 0°C, 5 ml of water, 5 ml of 15 wt.% NaOH solution, 0.5 ml of water and anhydrous MgSO ₄ were successively added. After stirring for 15 minutes, the mixture was filtered and concentrated to obtain 6.73 g (70%) of the expected product.

1H NMR (500 MHz, DMSOO δ ppm 4.34 (t, 1H), 3.89 (m, 1H), 3.48/3.44 (m+m, 2H), 3.24 (s, 3H), 3.24/3.21 (dd+ dd, 2H), 1.59/1.46 (m+m, 2H), 0.85 (s, 9H), 0.04/0.03 (s+s, 6H); HRMS-ERI (m/z): [M+H] ⁺ , calculated for C11H ₂₇ O ₃ Si: 235.1729; found: 235.1725.

Synthesis 2i: 4-[tert-butyl(dimethyl)silyl]oxy-3-methoxybutan-1-ol.

Step A: 3,4-dihydroxybutyl benzoate.

A mixture of 20.1 g but-3-enyl benzoate (113.5 mmol, 1 eq.), 21.8 g 4-methyl-4-oxidomorpholin-4ium hydrate (161.3 mmol, 1.42 eq.) and 2.83 g osmium tetroxide (2.5 wt.% in 2 -methylpropan-2-ole, 0.28 mmol, 0.0025 eq.) in 227 ml of 2-methylpropan-2-ol and 27 ml of water was stirred at 60°C for 18 hours. The reaction mixture was then quenched by adding at room temperature. 270 ml of 1 M Na ₂ S ₂ O ₃ solution, the mixture was diluted with DCM and the organic phase was washed with water and brine, dried and concentrated to obtain 22.22 g (93%) of 3,4-dihydroxybutyl benzoate.

- 32 046336 ¹ H NMR (500 MHz, DMSO-dg) δ ppm 7.97 (m, 2H), 7.65 (m, 1H), 7.53 (m, 2H), 4.68 (d, 1H), 4.59 (t, 1H), 4.39/4.34 (m+m, 2H), 3.63 (m, 1H), 3.37/3.30 (m+m, 2H), 1.93/1.64 (m+m, 2H); ¹³ C NMR (125 MHz, DMSOd6) δ ppm 166.2, 133.7, 129.5, 129.2, 68.5, 66.4, 62.5, 33.0; MSHR-ERI (m/z): [M+H] ⁺ , calculated for CnH ₁₅ O ₄ : 211.0970; found: 211.0971.

Step B: [4-[tert-butyl(dimethyl)silyl]oxy-3-hydroxybutyl]benzoate.

To 10 g of the product from step A (47.6 mmol, 1 eq.) and 6.52 g of imidazole (0.096 mol, 2 eq.) in 100 ml of DMF, 7.17 g of tert-butylchlorodimethylsilane (47.6 mmol, 1 eq.) was added in one portion. and the mixture was stirred for 1 hour. The mixture was then diluted with EtOAc, the organic phase was washed with conc. NaHCO ₃ , water and brine, dried over MgSO ₄ , filtered, and concentrated to give 14.1 g (91%) of the desired product.

1H NMR (400 MHz, DMSO^) δ ppm 7.95 (m, 2H), 7.64 (m, 1H), 7.51 (t, 2H), 4.76 (s, 1H), 4.38 (m, 2H), 3.65 (m, 1H), 3.56 (dd, 1H), 3.43 (dd, 1H), 1.96 (m, 1H), 1.65 (m, 1H), 0.84 (s, 9H), 0.02 (s, 6H).

Step C: [4-[tert-butyl(dimethyl)silyl]oxy-3-methoxybutyl]benzoate.

7.03 g of the product from step B (21.7 mmol, 1 eq.) and 13.9 g of H1,H1,III,III-tetramethylna(|)talin-1,8diamine (64.9 mmol, 3 eq.) were mixed in 420 ml of DCM, then in 8.0 g of trimethyloxonium tetrafluoroborate (54.1 mmol, 2.5 eq.) was added in one portion and the mixture was stirred at room temperature. within 18 hours. Complete transformation was observed. The mixture was filtered through a pad of celite, washed with DCM, then the organic phase was washed with water and brine, respectively. The solution was dried over MgSO ₄ , filtered, concentrated on celite and purified by flash column chromatography using heptane and EtOAc as eluents to obtain 4.03 g (55%) of the expected product.

1H NMR (400 MHz, DMSO^6) δ ppm 7.97 (m, 2H), 7.67 (tt, 1H), 7.52 (t, 2H), 4.35 (m, 2H), 3.63 (m, 2H), 3.35 (m, 1H), 3.33 (s, 3H), 1.94 (m, 1H), 1.80 (m, 1H), 0.85 (s, 9H), 0.03 (s, 6H).

Step D: 4-[tert-butyl(dimethyl)silyl]oxy-3-methoxybutan-1-ol.

To 4.0 g of the product from step C (11.83 mmol, 1 eq.) in 100 ml of a 1:1 mixture of THF and MeOH at 0°C was added 5.1 g of K ₂ CO ₃ (36.96 mmol, 3 eq.), and the mixture was stirred for 3 hours at 0°C and for 2 hours at room temperature. After adding conc. solution of NH4C1 and DCM at 0°C, the organic phase was washed with conc. NaHCO3 solution, water and brine, dried over MgSO4, filtered and concentrated to obtain 4.14 g (94%) of a 1:1 mixture of the target product and methyl benzoate.

1H NMR (400 MHz, DMSOA) δ ppm 4.38 (t, 1H), 3.57 (dd, 2H), 3.45 (q, 2H), 3.29 (s, 3H), 3.28 (m, 1H), 1.53 (m, 2H), 0.87 (s, 9H), 0.04 (s, 6H).

Synthesis of 2j: 3-[tert-butyl(dimethyl)silyl]oxy-2-methoxypropan-1-ol.

Step A: (2,2-dimethyl-1,3-dioxolan-4-yl)methyl benzoate.

To 20 g (2,2-dimethyl-1,3-dioxolan-4-yl)methanol (151 mmol, 1 eq.) and 25 ml N,N-diethylethanamine (182 mmol, 1.2 eq.) in 92 ml DCM at At 0°C, 21 mL of benzoyl chloride (182 mmol, 1.2 eq.) was added and the mixture was stirred at room temperature. for 18 hours. The reaction mixture was quenched with conc. NaHCO ₃ solution and the organic phase was washed with brine, dried and concentrated to obtain 37.0 g (98%) of the desired product.

1H NMR (400 MHz, DMSO<) δ ppm 7.98 (dd, 2H), 7.67 (m, 1H), 7.54 (t, 2H), 4.42 (m, 1H), 4.37 (dd, 1H), 4.27 (dd, 1H), 4.09 (dd, 1H), 3.82 (dd, 1H), 1.33 (s, 3H), 1.29 (s, 3H).

Step B: 2,3-dihydroxypropyl benzoate.

A mixture of 20 g of the product from stage A (80 mmol, 1 eq.), 220 ml of 1N. HCl solution and 220 ml EtOH were stirred at room temperature. for 18 hours. Then the reaction mixture was quenched with conc. Na2CO3 solution and concentrated, the residue was extracted with EtOAc. The combined organic phases were washed with brine and dried to obtain 15.48 g (98.7%) of the expected product.

1H NMR (400 MHz, DMSOD ₆ ) δ ppm 8.00 (dd, 2H), 7.67 (tt, 1H), 7.54 (t, 2H), 5.03 (d, 1H), 4.71 (t, 1H), 4.30 (dd, 1H), 4.17 (dd, 1H), 3.79 (sx, 1H), 3.45 (m, 2H).

Step C: [3-[tert-butyl(dimethyl)silyl]oxy-2-hydroxypropyl]benzoate.

To 5.62 g of the product from step B (28.7 mmol, 1 eq.) and 3.47 g of imidazole (51.0 mol, 2 eq.) in 50 ml of DMF, 4.04 g of tert-butylchlorodimethylsilane (26.8 mmol, 1 eq.) was added in one portion. and the mixture was stirred for 1 hour. The mixture was then diluted with EtOAc, the organic phase was washed with conc. NaHCO ₃ solution, water and brine, dried over MgSO ₄ , filtered, concentrated and purified by flash column chromatography using heptane and EtOAc as eluents to obtain 3.23 g (36%) of the desired product.

1H NMR (400 MHz, DMSOD ₆ ) δ ppm 7.99 (dd, 2H), 7.66 (tt, 1H), 7.53 (t, 2H), 5.13 (d, 1H), 4.31 (dd, 1H), 4.19 (dd, 1H), 3.83 (sx, 1H), 3.63 (m, 2H), 0.85 (s, 9H), 0.03 (s, 6H).

Step D: [3-[tert-butyl(dimethyl)silyl]oxy-2-methoxypropyl]benzoate.

3.0 g of the product from step C (9.6 mmol, 1 eq.) and 6.2 g of Yu.Yu.Sh.Sh-tetramethylnaphthalene-1,8-diamine (28.9 mmol, 3 eq.) were mixed in 160 ml of DCM, then added in one portion 3.54 g of trimethyloxonium tetrafluoroborate (23.9 mmol, 2.5 eq.), and the mixture was stirred at room temperature. for 18 hours. The mixture was filtered through a pad of celite, washed with DCM, then the organic phase was washed with water and

- 33 046336 saline solution, respectively. The solution was dried over MgSO ₄ , filtered, concentrated on celite and purified by flash column chromatography using heptane and EtOAc as eluents to obtain 1.7 g (55%) of the expected product.

1H NMR (400 MHz, DMSOHO δ ppm 7.97 (dd, 2H), 7.67 (tt, 1H), 7.54 (t, 2H), 4.44 (dd, 1H), 4.26 (dd, 1H), 3.72 (d , 2H), 3.57 (m, 1H), 3.38 (s, 3H), 0.86 (s, 9H), 0.05 (s, 3H), 0.04 (s, 3H).

Step E: 3-[tert-butyl(dimethyl)silyl]oxy-2-methoxypropan-1-ol.

To 1.68 g of the product from step D (5.17 mmol, 1 eq.) in 40 ml of a 1:1 mixture of THF and MeOH at 0°C was added 2.19 g of K ₂ CO ₃ (15.85 mmol, 3 eq.), and the mixture was stirred for 3 hours at 0°C and for 2 hours at room temperature. After adding conc. solution of NH ₄ Cl and DCM at 0°C, the organic phase was washed with conc. NaHCO ₃ solution, water and brine, dried over MgSO ₄ , filtered and concentrated to obtain 1.26 g (quantitative) of a 1:0.25 mixture of the target product and methyl benzoate.

¹ H NMR (400 MHz, DMSO<) δ ppm 4.54 (t, 1H), 3.63 (dd, 1H), 3.55 (dd, 1H), 3.39 (m, 2H), 3.32 (s, 3H), 3.15 (m, 1H), 0.86 (s, 9H), 0.03 (s, 6H).

Synthesis of 2k: 3-[tert-butyl(dimethyl)silyl]oxy-2-morpholinopropan-1-ol.

Step A: diethyl 2-morpholinopropanedioate.

To 16 g of morpholine (184 mmol, 1.6 eq.) and 30 g of K ₂ CO ₃ (217 mmol, 1.9 eq.) in 50 ml of MeCN, 27 g of diethyl bromomalonate (113 mmol, 1 eq.) was added and the exothermic reaction mixture was stirred in for 0.5 h. After cooling the mixture to rt, it was filtered and concentrated, and the crude product was purified by flash column chromatography using heptane and EtOAc as eluents to obtain 26.2 g (94%) of the expected product.

1H NMR (400 MHz, DMSOZH) δ ppm 4.27 (s, 1H), 4.16 (q, 4H), 3.57 (dd, 4H), 2.68 (dd, 4H), 1.20 (s, 6H).

Step B: 2-morpholinopropane-1,3-diol.

To 3.0 g of LiAlH ₄ (79 mmol, 4.8 eq.) in 60 ml of THF at 0°C was added 4.0 g of the product from step A (16.3 mmol, 1 eq.) in 25 ml of THF, and the mixture was stirred at 60°C for 4 hours. After cooling to room temperature, the mixture was treated with 10N. NaOH solution and the resulting precipitate was filtered off and washed with DCM. The phases were then separated, the aqueous phase was extracted with DCM and the combined organic phases were washed with brine, dried over MgSO4, filtered and concentrated to give 1.4 g (53%) of the expected product.

1H NMR (500 MHz, DMSOD ₆ ) δ ppm 4.28 (dd, 2H), 3.52 (m, 4H), 3.49/3.44 (m+m, 4H), 2.59 (m, 4H), 2.40 (qui, 1H); ¹³ C NMR (125 MHz, DMSOO δ ppm 67.5, 67.3, 59.1, 50.5; IR: 3373, 2856; HRMS-EI (m/z): [M] ⁺ , calculated for C ₇ H ₁₅ NO ₃ : 161.1052; found: 161.1053.

Step C: 3-[tert-butyl(dimethyl)silyl]oxy-2-morpholino-propan-1-ol.

To 1.32 g of the product from step B (8.22 mmol, 1 eq.) and 0.56 g of imidazole (8.24 mmol, 1 eq.) in 60 ml of DCM, 1.24 g of tert-butylchlorodimethylsilane (8.23 mmol, 1 eq.) in 10 ml was added dropwise DCM, and the mixture was stirred at room temperature. for 0.5 h. After addition of DCM, the solution was washed with water and brine, dried, concentrated and purified by flash column chromatography using heptane and EtOAc as eluents to obtain 0.79 g (35%) of the expected product.

1H NMR (500 MHz, DMSOHO δ ppm 4.30 (brt, 1H), 3.69/3.65 (dd+dd, 2H), 3.51 (m, 4H), 3.48/3.43 (m+m, 2H), 2.64/ 2.59 (m+m, 4H), 2.44 (m, 1H), 0.87 (s, 9H), 0.04/0.03 (s, 6H); ¹³ C NMR (125 MHz, DMSOL6) δ ppm 67.4, 67.4, 61.2, 59.2, 50.8, 26.3, 18.3, -5.0; IR: 2855; MSHR-EI (m/z): [M-C4H9] ⁺ , calculated for C9H20NO ₃ Si: 218.1217; found: 218.1202.

Synthesis 3a: methyl 5-[3-[4-[3-[tert-butoxycarbonyl(methyl)amino]prop-1-ynyl]-2fluorophenoxy]propyl]-2-[[5-[tert-butyl(dimethyl)silyl ]hydroxy-4-[tert-butyl(diphenyl)silyl]oxypentyl]amino]thiazole-4-carboxylate.

Step A: methyl 2-[tert-butoxycarbonyl-[5-[tert-butyl(dimethyl)silyl]oxy-4-[tert-butyl(diphenyl)silyl]oxypentyl]amino]-5-[3-[4-[3- [tert-butoxycarbonyl(methyl)amino]prop-1-ynyl]2-fluorophenoxy]propyl]thiazole-4-carboxylate.

Using the general Mitsunobu reaction procedure, starting with synthesis 1b as the appropriate carbamate and synthesis 2a as the appropriate alcohol, 2.5 g (61%) of the desired product was obtained.

1H NMR (500 MHz, DMSO<) δ ppm 7.60-7.33 (m, 10H), 7.28 (dd, 1H), 7.17 (m, 1H), 7.1 (t, 1H), 4.22 (s, 2H), 4.09 (t, 2H), 3.94 (m, 2H) , 3.71 (s, 3H), 3.67 (m, 1H), 3.38 (m, 2H), 3.22 (t, 2H), 2.85 (s, 3H), 2.07 (m, 2H), 1.65 (m, 2H), 1.48 (m, 2H), 1.45/1.40 (s+s, 18H), 0.93 (s, 9H), 0.71 (s, 9H), -0.17/-0.22 (s+s, 6H); ¹³ C NMR (125 MHz, DMSOZH) δ ppm. 147.4, 129, 119.3, 115.4, 85.1, 82.3, 73.3, 68.1, 65.6, 51.9, 46.5, 38.4, 33.8, 30.5, 30.5, 28.5/28, 27.2, 26.0, 23.1, 23.0 , -5.3; MSVR-ERI (m/z): [M+H] ⁺ , calculated for C ₅₅ H ₇₉ FN ₃ O ₉ SSi ₂ : 1032.5054; found: 1032.5060.

Step B: methyl 5-[3-[4-[3-[tert-butoxycarbonyl(methyl)amino]prop-1-ynyl]-2fluorophenoxy]propyl]-2-[[5-[tert-butyl(dimethyl)silyl ]hydroxy-4-[tert-butyl(diphenyl)silyl]oxypentyl]amino]thiazole-4-carboxylate.

Using the general HFIP deprotection procedure, starting with the product from Step A as the appropriate carbamate, 1.2 g (53%) of the desired product was obtained.

- 34 046336 ¹ H NMR (500 MHz, DMSO-dg) δ ppm 7.68-7.35 (m, 10H), 7.56 (t, 1H), 7.30 (d, 1H), 7.20 (d, 1H), 7.11 (t, 1H), 4.22 (br., 2H), 4.07 (t, 2H ), 3.70 (m, 1H), 3.68 (s, 3H), 3.42/3.38 (dd+dd, 2H), 3.11 (t, 2H), 3.04 (brq., 2H), 2.86 (brq., 3H), 1.99 (quint., 2H), 1.54 (m, 2H), 1.53/1.45 (m+m, 2H), 1.41 (s, 9H), 0.97 (s, 9H), 0.74 (s, 9H), 0.14/- 0.18 (s+s, 6H); ¹³ C NMR (125 MHz, DMSOL) δ ppm 164.6, 163.0, 154.9, 151.4, 147.5, 136.9, 136.0, 129.1, 119.3, 115.4, 114.8, 85.2, 82.3, 79.8, 73.6, 68.0, 66.2, 51.7, 44.7 , 38.5, 33.8, 31.1, 30.6, 28.5, 27.2, 26.2, 24.3, 23.3, 19.4, 18.3, -5.2; MSVR-ERI (m/z): [M+H] ⁺ , calculated for C ₅₀ H ₇ iFN ₃ O ₇ SSi ₂ : 932.4530; found: 932.4526.

Synthesis 3b: methyl 2-[[5-tert-butyl(dimethyl)silyl]oxy-4-tert-butyl(diphenyl)silyl]oxypentyl]amino]-5-[3-[4-[3-(dimethylamino)prop -1-ynyl]-2-fluorophenoxy]propyl]thiazole-4-carboxylate.

Step A: methyl 2-[tert-butoxycarbonyl-[5-[tert-butyl(dimethyl)silyl]oxy-4-[tert-butyl(diphenyl)silyl]oxypentyl]amino]-5-[3-[4-[3 - (dimethylamino)prop-1-ynyl]-2 fluorophenoxy]propyl]thiazole-4-carboxylate.

Using the general Mitsunobu reaction procedure, starting with synthesis 1c as the appropriate carbamate and synthesis 2a as the appropriate alcohol, 3.2 g (65%) of the desired product was obtained.

1H NMR (500 MHz, DMSOHO δ ppm 7.60-7.35 (m, 10H), 7.28 (dd, 1H), 7.18 (m, 1H), 7.10 (t, 1H), 4.10 (t, 2H), 3.95 (m, 2H), 3.72 (s, 3H), 3.68 (m, 1H), 3.41 (s, 2H), 3.38 (m, 2H), 3.22 (t, 2H), 2.21 (s, 6H), 2.07 ( m, 2H), 1.66 (m, 2H), 1.45 (s, 9H), 1.45 (m, 2H), 0.94 (s, 9H), 0.72 (s, 9H), -0.16/-0.21 (s+s, 6H); ¹³ C NMR (125 MHz, DMSOO δ ppm 162.8, 147.2, 128.9, 119.2, 115.4, 84.2, 73.3, 68.1, 65.6, 52.0, 48.2, 46.7, 44.3, 30.7, 30. 5, 28.0, 27.2, 26.1, 23.1, 23.0, -5.3; MSHR-ERI (m/z): [M+H] ⁺ , calculated for C51H ₇ 3FN3O ₇ SSi2: 946.4686; found: 946.4684.

Step B: methyl 2-[[5-[tert-butyl(dimethyl)silyl]oxy-4-[tert-butyl(diphenyl)silyl]oxypentyl]amino]-5-[3-[4-[3-(dimethylamino )prop-1-ynyl]-2-fluorophenoxy]propyl]thiazole-4-carboxylate.

Using the general HFIP deprotection procedure, starting with the product from Step A as the appropriate carbamate, 2.2 g (55%) of the desired product was obtained.

1H NMR (500 MHz, DMSOO δ ppm 7.64-7.37 (m, 10H), 7.56 (t, 1H), 7.28 (dd, 1H), 7.18 (m, 1H), 7.10 (t, 1H), 4.07 (t, 2H), 3.69 (q, 1H), 3.69 (s, 3H), 3.41 (s, 2H), 3.41 (m, 2H), 3.11 (t, 2H), 3.04 (q, 2H), 2.21 ( s, 6H), 2.00 (m, 2H), 1.55 (m, 2H), 1.54/1.45 (m+m, 2H), 0.97 (s, 9H), 0.74 (s, 9H), -0.14/-0.17 ( s+s, 6H); ¹³ C NMR (125 MHz, DMSOO δ ppm 164.6, 147.2, 129.0, 119.2, 115.4, 73.6, 68.0, 66.2, 51.7, 48.2,

44.8, 44.3, 31.1, 30.7, 27.2, 26.2, 24.4, 23.3, -5.2; MSHR-ERI (m/z): [M+H] ⁺ , calculated for C46H66FN3OsSSi2: 846.4162; found: 846.4160.

Synthesis 3c: methyl 5-[3-[4-[3-[tert-butoxycarbonyl(methyl)amino]prop-1-ynyl]-2fluorophenoxy]propyl]-2-[4-tert-butyl(dimethyl)silyl]oxybutylamino ]thiazole-4-carboxylate.

Step A: Methyl 2-[tert-butoxycarbonyl-[4-[tert-butyl(dimethyl)silyl]oxybutyl]amino]-5-[3-(2fluoro-4-iodophenoxy)propyl]thiazole-4-carboxylate.

Using the general alkylation procedure, starting with 5.36 g of synthesis 1a (10 mmol, 1 eq.) and 3.77 g of tert-butyl-(4-iodobutoxy)-dimethylsilane (3.10 ml, 12 mmol, 1.2 eq.) as a suitable halide, 6.1 g (84%) of the target product were obtained.

1H NMR (500 MHz, DMSOO δ ppm 7.58 (dd, 1H), 7.45 (m, 1H), 6.97 (t, 1H), 4.06 (t, 2H), 4.01 (t, 2H), 3.75 (s , 3H), 3.58 (t, 2H), 3.21 (t, 2H), 2.06 (m, 2H), 1.68 (m, 2H), 1.50 (s, 9H), 1.43 (m, 2H), 0.82 (s, 9H), -0.01 (s, 6H); ¹³ C NMR (125 MHz, DMSO-bb) δ ppm 147.0, 134.0, 124.9, 117.6, 68.2, 62.6, 52.1, 46.4, 30.5, 30.0, 28.1, 26.2 , 24.5, 23.2, -4.9; MSHR-ERI (m/z): [M+H] ⁺ , calculated for C ₂ 9H ₄₅ N ₂ O ₆ FSiSI: 723.1791; found: 723.1775.

Step B: Methyl 2-[4-[tert-butyl(dimethyl)silyl]oxybutylamino]-5-[3-(2-fluoro-4iodophenoxy)propyl]thiazole-4-carboxylate.

Using the general HFIP deprotection procedure, starting with 6.0 g of the product from Step A (8.30 mmol) as the appropriate carbamate, 3.0 g (58%) of the desired product was obtained.

1H NMR (500 MHz, DMSO^ ₆ ) δ ppm 7.59 (dd, 1H), 7.58 (t, 1H), 7.45 (dm, 1H), 6.97 (t, 1H), 4.03 (t, 2H), 3.69 (s, 3H), 3.58 (t, 2H), 3.16 (q, 2H), 3.11 (t, 2H), 1.98 (m, 2H), 1.59-1.44 (m, 4H), 0.84 (s, 9H), 0.01 (s, 6 H); ¹³ C NMR (125 MHz, DMSOO δ ppm 164.7, 163.0, 152.1, 147.0, 136.9, 136.1, 134.0, 124.9, 117.6, 82.4, 68.0, 62.7, 51.7, 44.3, 30. 6, 30.6, 26.3, 25.6, 23.3 , -4.8; MSHR-ERI (m/z): [M+H] ⁺ , calculated for C ₂₄ H ₃₇ FIN ₂ O ₄ SSi: 623.1266; found: 623.1272.

Step C: methyl 5-[3-[4-[3-[tert-butoxycarbonyl(methyl)amino]prop-1-ynyl]-2fluorophenoxy]propyl]-2-[4-[tert-butyl(dimethyl)silyl] hydroxybutylamino]thiazole-4-carboxylate.

Using the general Sonogashira reaction procedure, starting with 3.00 g of the product from step B (4.82 mmol, 1 eq.) and 1.63 g of tert-Butyl N-methyl-N-prop-2-ynylcarbamate (9.63 mmol, 2 eq.) as the appropriate acetylene , 2.50 g (65%) of the target product were obtained.

1H NMR (500 MHz, DMSOO δ ppm 7.58 (t, 1H), 7.31 (brd., 1H), 7.21 (d, 1H), 7.13 (t, 1H), 4.23 (br., 2H), 4.07 (t, 2H), 3.69 (s, 3H), 3.58 (t, 2H), 3.16 (q, 2H), 3.11 (t, 2H), 2.86 (br., 3H), 2 (quint., 2H), 1.54 (m, 2H), 1.49 (m, 2H), 1.41 (s, 9H), 0.84 (s, 9H), 0.01 (s, 6H); ¹³ C NMR (125 MHz, DMSOO δ ppm 129.1, 119.3, 115.4, 68.0, 62.7, 51.7, 44.3, 38.6, 33.8, 30.6, 30.1, 28.5, 26.3, 25.6, 23.3, -4.8; MSVR-ERI (m/z): [M+H] ⁺ ,

- 35 046336 calculated for C ₃₃ H ₅₁ FN ₃ O ₆ SSi: 664.3246; found: 664.3245.

Synthesis 3d: methyl 2-[4-[tert-butyl(dimethyl)silyl]oxybutylamino]-5-[3-[4-[3-(dimethylamino)prop1-ynyl]-2-fluorophenoxy]propyl]thiazol-4- carboxylate.

Using the general Sonogashira reaction procedure, starting from 3.00 g of synthesis product 3c, step B (4.82 mmol, 1 eq.), and 801 mg of KH-dimethyl-N-prop-2-yne-1-amine (9.63 mmol, 2 eq. .) as a suitable acetylene, 2.20 g (79%) of the target product were obtained.

¹ H NMR (500 MHz, DMSOO δ ppm 7.58 (t, 1H), 7.30 (dd, 1H), 7.20 (dm, 1H), 7.12 (t, 1H), 4.07 (t, 2H), 3.69 ( s, 3H), 3.58 (t, 2H), 3.44 (s, 2H), 3.16 (q, 2H), 3.12 (t, 2H), 2.24 (s, 6H), 2.00 (m, 2H), 1.60-1.44 (m, 4H), 0.84 (s, 9H), 0.01 (s, 6H); ¹³ C NMR (125 MHz, DMSO^) δ ppm 129.0, 119.2, 115.4, 84.9, 84.3, 68.0, 62.7, 51.7 , 48.1, 44.3, 44.2, 30.6, 30.2, 26.3, 25.6, 23.3, -4.8; MSHR-ERI (m/z): [M+H]+, calculated for C ₂₉ H ₄₅ FN ₃ O ₄ SSi: 578.2878; found: 578.2865.

Synthesis 3e: ethyl 5-(3-chloropropyl)-2-(methylamino)thiazole-4-carboxylate.

A suspension of 2.25 g of methylthiourea (25.0 mmol, 1 eq.) in 100 ml of ethanol was cooled to 0°C, and then at this temperature 7.46 g of ethyl 3-bromo-6-chloro-2-oxohexanoate (27.5 mmol, 1.1 eq. .). After stirring for 15 min at 0°C, 7 mL of TEA (5.06 g, 50 mmol, 2 eq.) was added. The resulting mixture was stirred overnight at room temperature. We observed a complete transformation. Volatiles were removed in vacuo, then the resulting residue was partitioned between EtOAc and water. The layers were separated, then the organic layer was washed with water, followed by washing with brine. The combined organic layers were dried over Na ₂ SO ₄ , filtered and the filtrate was concentrated under reduced pressure. The crude product was then purified by flash column chromatography using heptane and EtOAc as eluents to obtain 5 g (76%) of the desired product.

¹ H NMR (400 MHz, DMSO^) δ ppm 7.55 (q, 1H), 4.21 (q, 2H), 3.65 (t, 2H), 3.09 (m, 2H), 2.78 (d, 3H), 1.98 (m, 2H), 1.26 (t, 3H); ¹³ C NMR (125 MHz, DMSO^) δ ppm 165.6, 162.5, 137.4, 135.5, 60.5, 45.0, 34.1, 31.2, 24.4, 14.7; MSHR-ERI (m/z): [M+H] ⁺ , calculated for C ₁₀ H ₁₆ ClN ₂ O ₂ S: 263.0616; found: 263.0615.

Synthesis 3f: methyl 5-[3-[4-[3-[tert-butoxycarbonyl(methyl)amino]prop-1-ynyl]-2fluorophenoxy]propyl]-2-[[5-tert-butyl(dimethyl)silyl] hydroxy-4-methoxypentyl]amino]thiazole-4carboxylate.

Step A: methyl 2-[tert-butoxycarbonyl-[5-[tert-butyl(dimethyl)silyl]oxy-4-[tert-butyl(diphenyl)silyl]oxypentyl]amino]-5-[3-[4-[3- [tert-butoxycarbonyl(methyl)amino]prop-1-ynyl]2-fluorophenoxy]propyl]thiazole-4-carboxylate.

Using the general Mitsunobu reaction procedure, starting with 577 mg of synthesis 1b (1 mmol) as the appropriate carbamate and 496 mg of synthesis 2b (2 mmol) as the appropriate alcohol, 790 mg (96%) of the desired product was obtained.

Step B: methyl 5-[3-[4-[3-[tert-butoxycarbonyl(methyl)amino]prop-1-ynyl]-2fluorophenoxy]propyl]-2-[[5-[tert-butyl(dimethyl)silyl ]oxy-4-methoxypentyl]amino]thiazole-4carboxylate.

Using the general HFIP deprotection procedure, starting with 790 mg of the product from Step A (0.95 mmol) as the appropriate carbamate, 270 mg (38%) of the desired product was obtained.

¹ H NMR (500 MHz, DMSOD6) δ ppm 7.58 (t, 1H), 7.31 (brd, 1H), 7.21 (dm, 1H), 7.13 (t, 1H), 4.23 (brs, 2H), 4.07 (t, 2H), 3.69 (s, 3H), 3.56 /3.53 (dd+dd, 2H), 3.29 (s, 3H), 3.16 (m, 1H), 3.15 (m, 2H), 3.11 (t, 2H), 2.86 (brs, 3H), 2.00 (m, 2H ), 1.63-1.43 (m, 4H), 1.42 (s, 9H), 0.84 (s, 9H), 0.02 (s, 6H); MSVR-ERI (m/z): [M+H] ⁺ , calculated for C35H ₅₅ FN ₃ O ₇ SSi: 708.3508; found: 708.3502.

Synthesis of 3g: methyl 5-[3-[4-[3-(tert-butoxycarbonylamino)prop-1-ynyl]-2-fluorophenoxy]propyl]-2[4-[tert-butyl(dimethyl)silyl]oxybutylamino]thiazole -4-carboxylate.

Using the general Sonogashira reaction procedure, starting with 880 mg of synthesis product 3c, step B (1.41 mmol, 1 eq.), and 438 mg tert-butyl Nprop-2-ynylcarbamate (2.82 mmol, 2 eq.) as the appropriate acetylene , 918 mg (85%) of the target product were obtained.

¹ H NMR (500 MHz, DMSOD6) δ ppm 7.58 (t, 1H), 7.34 (t, 1H), 7.26 (ff, 1H), 7.17 (m, 1H), 7.12 (t, 1H), 4.06 (t, 2H), 3.95 (d, 2H), 3.69 (s, 3H), 3.58 (t, 2H), 3.17 (q, 2H), 3.11 (t, 2H), 2.00 (m, 2H), 1.54 (m, 2H), 1.50 (m, 2H), 1.39 ( s, 9H), 0.85 (s, 9H), 0.01 (s, 6H); ¹³ C NMR (125 MHz, DMSO^) δ ppm 164.7, 163.0, 147.4, 136.9, 136.1, 129.0, 119.1, 115.4, 68.0, 62.7, 51.7, 44.3, 30.6, 30.5, 30.2, 28.7, 26.3, 25.6, 23.3, 1 8.4, -4.8 MSVR-ERI (m/z) : [M+H] ⁺ , calculated for C ₃₂ H ₄ 9FN ₃ O ₆ SSi: 650.3090; found: 650.3093.

Synthesis 3h: methyl 5-[3-[4-[3-[tert-butoxycarbonyl(methyl)amino]prop-1-ynyl]-2fluorophenoxy]propyl]-2-[2-(2,2-dimethyl-1, 3-dioxolan-4-yl)ethylamino]thiazol-4-carboxylate.

Step A: methyl 2-[tert-butoxycarbonyl-[2-(2,2-dimethyl-1,3-dioxolan-4-yl)ethyl]amino]-5-[3-(2fluoro-4-iodophenoxy)propyl] thiazole-4-carboxylate.

Using the general Mitsunobu reaction procedure, starting from 2.68 g of synthesis product 1a (5 mmol, 1 eq.) and 1.46 g of 2-(2,2-dimethyl-1,3-dioxolan-4-yl)ethanol (1.42 ml, 10 mmol, 2 eq.) as a suitable alcohol, 2.8 g (84%) of the desired product were obtained.

¹ H NMR (500 MHz, DMSOO δ ppm 7.57 (dd, 1H), 7.44 (dm, 1H), 6.96 (t, 1H), 4.12/4.02 (m+m, 2H),

- 36 046336

4.07 (m, 1H), 4.05 (t, 2H), 4.02/3.54 (dd+dd, 2H), 3.75 (s, 3H), 3.21 (t, 2H), 2.06 (m, 2H), 1.86/1.82 ( m+m, 2H), 1.51 (s, 9H), 1.29 (s, 3H), 1.22 (s, 3H); ¹³ C NMR (125 MHz, DMSO-de) δ ppm 134.0, 124.9, 117.6, 73.8, 68.9, 68.1, 52.0, 44.0, 32.2, 30.5, 28.1, 27.3, 25.9, 23.1; MSVR-ERI (m/z): [M+H] ⁺ , calculated for C26H35FIN2O7S: 665.1188; found: 665.1175.

Step B: Methyl 2-[2-(2,2-dimethyl-1,3-dioxolan-4-yl)ethylamino]-5-[3-(2-fluoro-4iodophenoxy)propyl]thiazole-4-carboxylate.

Using the general HFIP deprotection procedure, starting with 2.5 g of the product from Step A (3.80 mmol) as the appropriate carbamate, 1.6 g (75%) of the desired product was obtained.

1H NMR (500 MHz, DMSOO δ ppm 7.6 (t, 1H), 7.59 (dd, 1H), 7.45 (dm, 1H), 6.97 (dd, 1H), 4.10 (m, 1H), 4.03 (t , 2H), 4.01/3.48 (dd+dd, 2H), 3.69 (s, 3H), 3.27/3.19 (m+m, 2H), 3.11 (t, 2H), 1.99 (m, 2H), 1.76/1.72 (m+m, 2H), 1.31 (s, 3H), 1.25 (s, 3H); HRMS-ERI (m/z): [M+H] ⁺ , calculated for C ₂₁ H ₂₇ FIN ₂ O ₅ S: 565.0663; found: 565.0642.

Step C: methyl 5-[3-[4-[3-[tert-butoxycarbonyl(methyl)amino]prop-1-ynyl]-2fluorophenoxy]propyl]-2-[2-(2,2-dimethyl-1, 3-dioxolan-4-yl)ethylamino]thiazol-4-carboxylate.

Using the general Sonogashira reaction procedure, starting with 400 mg of the product from step B (0.71 mmol, 1 eq.) and 240 mg of tert-butyl N-methyl-N-prop-2-ynylcarbamate (1.42 mmol, 2 eq.) as suitable acetylene, 300 mg (70%) of the target product were obtained.

1H NMR (500 MHz, DMSOO δ ppm 7.60 (t, 1H), 7.31 (brd, 1H), 7.21 (dd, 1H), 7.13 (t, 1H), 4.23 (brs, 2H), 4.09 (m , 1H), 4.07 (t, 2H), 4.00/3.48 (dd+dd, 2H), 3.69 (s, 3H), 3.27/3.19 (m+m, 2H), 3.12 (t, 2H), 2.86 (brs , 3H), 2.00 (m, 2H), 1.74 (m, 2H), 1.41 (s, 9H), 1.31 (s, 3H), 1.25 (s, 3H); ¹³ C NMR (125 MHz, DMSOO δ m. d. 164.5, 136.9, 136.4, 129.1, 119.3, 115.4, 85.2, 82.3, 73.8, 69.0, 68.0, 51.7, 41.4, 38.4, 33.8, 33.2, 30.6, 28.5, 27.3, 2 6.1, 23.3; MSVR-ERI (m/ z): [M+H] ⁺ , calculated for C ₃₀ H ₄₁ FN ₃ O ₇ S: 606.2644; found: 606.2650.

Synthesis 3i: methyl 5-[3-[4-[3-(dimethyl)amino]prop-1-ynyl]-2-fluorophenoxy]propyl]-2-[2-(2,2dimethyl-1,3-dioxolane- 4-yl)ethylamino]thiazole-4-carboxylate.

Using the general Sonogashira reaction procedure, starting from 400 mg of synthesis product 3h, step B (0.71 mmol, 1 eq.), and 117 mg of N-dimethylprop-2-yn-1-amine (1.42 mmol, 2 eq.) in as suitable acetylene, 250 mg (58%) of the desired product were obtained.

1H NMR (500 MHz, DMSOO δ ppm 7.61 (t, 1H), 7.29 (dd, 1H), 7.2 (dd, 1H), 7.12 (t, 1H), 4.1 (m, 1H), 4.07 (t , 2H), 4/3.48 (dd+dd, 2H), 3.69 (s, 3H), 3.41 (s, 2H), 3.28/3.19 (m+m, 2H), 3.12 (t, 2H), 2.22 (s , 6H), 2 (qn, 2H), 1.75/1.72 (m+m, 2H), 1.31 (s, 3H), 1.25 (s, 3H); ¹³ C NMR (125 MHz, DMSOO δ ppm 164.5 , 163, 151.5, 147.3, 136.9, 136.4, 128.9, 119.2, 115.4, 115.3, 108.4, 85.2, 84.2, 73.8, 69.0, 68.0, 51.8, 48.1, 44.3, 41.4, 33.3, 30.6, 27.4, 26.1, 23.4; MSVR -ERI (m/z): [M+H] ⁺ , calculated for C26H35FN3O5S: 520.2281; found: 520.2272.

Synthesis 3j: methyl 2-[2-(2,2-dimethyl-1,3-dioxolan-4-yl)ethylamino]-5-[3-[2-fluoro-4-[3-[(4methoxyphenyl)methylmethylamino] prop-1-ynyl]phenoxy]propyl]thiazole-4-carboxylate.

Using the general Sonogashira reaction procedure, starting from 400 mg of synthesis product 3h, step B (0.71 mmol, 1 eq.), and 268 mg of Na[(4-methoxyphenyl)methyl]-N-methyl-prop-2-yn-1 -amine (1.42 mmol, 2 eq.) as the appropriate acetylene gave 280 mg (63%) of the desired product.

1H NMR (500 MHz, DMSO-de) δ ppm 7.61 (t, 1H), 7.33 (dd, 1H), 7.24 (dd, 1H), 7.23 (d, 2H), 7.13 (t, 1H), 6.89 (d, 2H), 4.10 (qn, 1H), 4.07 (t, 2H), 4/3.48 (dd+dd, 2H), 3.73 (s, 3H), 3.7 (s, 3H), 3.49 (s, 2H), 3.44 (s, 2H), 3.28/3.19 (m +m, 2H), 3.13 (t, 2H), 2.24 (s, 3H), 2.01 (qn, 2H), 1.75/1.73 (m+m, 2H), 1.31 (s, 3H), 1.25 (s, 3H ); ¹³ C NMR (125 MHz, DMSO-de) δ ppm 164.5, 163.0, 158.8, 151.5, 147.3, 136.6, 136.5, 130.8, 130.5, 129.0, 119.3, 115.4, 115.3, 114.1, 108.4, 84.9, 84.5, 73.8, 69.0, 68.0, 59.4, 55.5, 51.8, 45.8, 41.7, 41.4, 33.3, 30.6, 27.3, 26.1, 23.4; MSHR-ERI (m/z): [M+H] ⁺ , calculated for C ₃₃ H ₄₁ FN ₃ O ₆ S: 626.2694; found: 626.2697.

Synthesis 3k: methyl 2-[2-(2,2-dimethyl-1,3-dioxolan-4-yl)ethylamino]-5-[3-[2-fluoro-4-[3-[methyl(ptolylsulfonyl)amino ]prop-1-ynyl]phenoxy]propyl]thiazole-4-carboxylate.

Using the general Sonogashira reaction procedure, starting from 400 mg of the product of synthesis 3h, step B (0.71 mmol, 1 eq.), and 316 mg of N4-dimethyl-N-prop-2-ynylbenzenesulfonamide (1.42 mmol, 2 eq.) in as suitable acetylene, 260 mg (55%) of the desired product were obtained.

1H NMR (500 MHz, DMSOO δ ppm 7.72 (d, 2H), 7.61 (t, 1H), 7.4 (d, 2H), 7.07 (t, 1H), 6.88 (dd, 1H), 6.86 (dd , 1H), 4.25 (s, 2H), 4.1 (qn, 1H), 4.05 (t, 2H), 4/3.48 (dd+dd, 2H), 3.69 (s, 3H), 3.27/3.19 (m+m , 2H), 3.11 (t, 2H), 2.79 (s, 3H), 2.33 (s, 3H), 1.99 (qn, 2H), 1.75/1.73 (m+m, 2H), 1.31 (s, 3H), 1.25 (s, 3H); ¹³ C NMR (125 MHz, DMSOO δ ppm 164.5, 163.0, 151.2, 147.5, 144.0, 136.9, 136.4, 134.4, 130.2, 128.9, 128.2, 119.1, 115.2, 114.2, 108.4, 84.6, 81.9, 73.9, 69.0, 68.1, 51.8, 41.5, 40.4, 34.9, 33.3, 30.6, 27.4, 26.1, 23.4, 21.4; MSVRERI (m/z): [M+H] ⁺ , calculated for C ₃₂ H ₃₉ FN ₃ O ₇ S ₂ : 660.22079; found: 660.2231.

Synthesis 31: methyl 5-[3-[4-[3-[tert-butoxycarbonyl(methyl)amino]prop-1-ynyl]-2fluorophenoxy]propyl]-2-[[3-tert-butyl(dimethyl)silyl] hydroxy-4-methoxybutyl]amino]thiazole-4carboxylate.

Step A: methyl 2-[tert-butoxycarbonyl-[3-[tert-butyl(dimethyl)silyl]oxy-4-methoxybutyl]amino]-5-[3-[4-[3-[tert-butoxycarbonyl(methyl) amino]prop-1-ynyl]-2-fluorophenoxy]propyl]thiazole-4carboxylate.

- 37 046336

Using the general Mitsunobu reaction procedure, starting with 577 mg of synthesis 1b (1 mmol, 1 eq) as the appropriate carbamate and 469 mg of synthesis 2h (2 mmol, 2 eq) as the appropriate alcohol, gave 794 mg (99 %) of the target product.

Step B: methyl 5-[3-[4-[3-[tert-butoxycarbonyl(methyl)amino]prop-1-ynyl]-2fluorophenoxy]propyl]-2-[[3-[tert-butyl(dimethyl)silyl ]oxy-4-methoxybutyl]amino]thiazole-4carboxylate.

Using the general HFIP deprotection procedure, starting from 794 mg of the product from Step A (1 mmol) as the appropriate carbamate, 320 mg (46%) of the desired product was obtained.

1H NMR (400 MHz, DMSOD ₆ ) δ ppm 7.54 (t, 1H), 7.28 (d, 1H), 7.18 (d, 1H), 7.09 (t, 1H), 4.04 (t, 2H), 4.20 (bs, 2H), 3.89-3.81 (m, 1H) , 3.67 (s, 3H), 3.23-3.14 (m, 4H), 3.22 (s, 3H), 3.10 (t, 2H), 2.83 (brs, 3H), 2.031.93 (m, 2H), 1.74-1.50 (m, 2H), 1.39 (s, 9H), 0.81 (s, 9H), 0.00 (s, 3H), -0.01 (s, 3H).

Synthesis of 3m: methyl 5-[3-[4-[3-[tert-butoxycarbonyl(methyl)amino]prop-1-ynyl]-2fluorophenoxy]propyl]-2-[5-[tert-butyl(dimethyl)silyl] hydroxypentylamino]thiazole-4-carboxylate.

Step A: Methyl 2-[tert-butoxycarbonyl-[5-[tert-butyl(dimethyl)silyl]oxypentyl]amino]-5-[3(2-fluoro-4-iodophenoxy)propyl]thiazole-4-carboxylate.

Using the general alkylation procedure, starting from 5.00 g of synthesis 1a (7.55 mmol, 1 eq.) and 2.97 g of tert-butyl-(5-iodopentoxy)-dimethylsilane (9.06 mmol, 1.2 eq.) as the appropriate halide, gave 4.72 g (85%) of the target product.

1H NMR (500 MHz, DMSOO δ ppm 7.59 (dm, 1H), 7.46 (dm, 1H), 6.99 (t, 1H), 4.07 (t, 2H), 4.01 (t, 2H), 3.76 (s , 3H), 3.56 (t, 2H), 3.21 (t, 2H), 2.07 (m, 2H), 1.64 (m, 2H), 1.52 (s, 9H), 1.48 (m, 2H), 1.30 (m, 2H), 0.82 (s, 9H), -0.02 (s, 6H); ¹³ C NMR (125 MHz, DMSOO δ ppm 163.1, 156.3/153.2, 151.8, 147.0, 143.2, 135.6, 133.9, 124.9, 117.6 , 83.5, 82.3, 67.3, 62.5, 52.0, 46.5, 32.3, 30.5, 28.1, 27.6, 26.3, 22.9, 22.6, 18.1, -4.9; MSVR-ERI (m/z): [M+H]+, calculated for C ₃₀ H ₄₇ FIN ₂ O ₆ SSi: 737.1947; found: 737.1948.

Step B: methyl 2-[[5-[tert-butyl(dimethyl)silyl]oxypentyl]amino]-5-[3-(2-fluoro-4iodophenoxy)propyl]thiazole-4-carboxylate.

A general HFIP deprotection procedure was used, starting with 2.36 g of the product from Step A (3.20 mmol) as the appropriate carbamate. After completion of the reaction, the reaction mixture was evaporated to dryness under reduced pressure to obtain crude target product.

1H NMR (400 MHz, DMSOHO δ ppm 7.60-7.54 (m, 2H), 7.44 (d, 1H), 6.96 (t, 1H), 4.03 (t, 2H), 3.69 (s, 3H), 3.56 (t, 2H), 3.18-3.08 (m, 4H), 1.98 (qv., 2H), 1.55-1.42 (m, 4H), 1.37-1.29 (m, 2H), 0.84 (s, 9H), 0.00 ( s, 6H).

Step C: methyl 5-[3-[4-[3-[tert-butoxycarbonyl(methyl)amino]prop-1-ynyl]-2fluorophenoxy]propyl]-2-[5-[tert-butyl(dimethyl)silyl] hydroxypentylamino]thiazole-4-carboxylate.

Using the general Sonogashira reaction procedure, starting from the crude product from step B and tert-butyl N-methyl-N-prop-2-ynylcarbamate as the appropriate acetylene, 1.92 g (89% over 2 steps) of the desired product was obtained.

1H NMR (500 MHz, DMSO<) δ ppm 7.57 (br., 1H), 7.31 (d, 1H), 7.21 (d, 1H), 7.12 (t, 1H), 4.23 (br., 2H), 4.07 (t, 2H), 3.69 (s, 3H) , 3.56 (t, 2H), 3.14 (m, 2H), 3.11 (t, 2H), 2.86 (br., 3H), 2.00 (quint., 2H), 1.51 (m, 2H), 1.45 (m, 2H), 1.41 (s, 9H), 1.33 (m, 2H), 0.84 (s, 9H), 0.00 (s, 6H); ¹³ C NMR (125 MHz, DMSO^) δ ppm 129.1, 119.3, 115.4, 68.0, 62.8, 51.7, 44.5, 38.6, 33.8, 32.4, 30.6, 28.9, 28.5, 26.3, 23.3, 23.2, -4.9; MSHR-ERI (m/z): [M+H]+, calculated for C ₃₄ H ₅₃ FN ₃ O ₆ SSi: 678.3403; found: 678.3393.

Synthesis of 3n: methyl 5-[3-[4-[3-[tert-butoxycarbonyl(methyl)amino]prop-1-ynyl]-2fluorophenoxy]propyl]-2-[3-tert-butyl(dimethyl)silyl]oxypropylamino ]thiazole-4-carboxylate.

Step A: methyl 2-[tert-butoxycarbonyl-[3-[tert-butyl(dimethyl)silyl]oxypropyl]amino]-5-[3[4-[3-[tert-butoxycarbonyl(methyl)amino]prop-1 -inyl]-2-fluorophenoxy]propyl]thiazole-4-carboxylate.

Using the general Mitsunobu reaction procedure, starting with 577 mg of synthesis 1b (1 mmol, 1 eq.) as the appropriate carbamate and 380 mg of 3-[tert-butyl(dimethyl)silyl]oxypropan-1ol (2 mmol, 2 eq. ) as a suitable alcohol, 600 mg (80%) of the desired product were obtained.

Step B: methyl 5-[3-[4-[3-[tert-butoxycarbonyl(methyl)amino]prop-1-ynyl]-2fluorophenoxy]propyl]-2-[3-[tert-butyl(dimethyl)silyl] hydroxypropylamino]thiazole-4-carboxylate.

Using the general HFIP deprotection procedure, starting with the product from Step A as the suitable carbamate, 310 mg (47%) of the desired product was obtained.

1H NMR (400 MHz, DMSOD ₆ ) δ ppm 7.50 (t, 1H), 7.30 (d, 1H), 7.20 (d, 1H), 7.11 (t, 1H), 4.21 (bs, 2H), 4.05 (t, 2H), 3.62 (t, 2H), 3.67 (s, 3H), 3.19 (q, 2H), 3.10 (t, 2H), 2.84 (brs, 3H), 2.04-1.94 (m, 2H), 1.74-1.63 (m, 2H), 1.40 (s, 9H ), 0.84 (s, 9H), 0.00 (s, 6H).

Synthesis of 3o: methyl 5-[3-[4-[3-[tert-butoxycarbonyl(methyl)amino]prop-1-ynyl]-2fluorophenoxy]propyl]-2-[(2,2-dimethyl-1,3- dioxolan-4-yl)methylamino]thiazol-4-carboxylate.

Step A: methyl 2-[tert-butoxycarbonyl-[(2,2-dimethyl-1,3-dioxolan-4-yl)methyl]amino]-5-[3-[4[3-[tert-butoxycarbonyl(methyl )amino]prop-1-ynyl]-2-fluorophenoxy]propyl]thiazole-4-carboxylate.

Using the general Mitsunobu reaction procedure, starting with 577 mg of synthesis 1b (1 mmol, 1 eq.) as the appropriate carbamate and 264 mg of (2,2-dimethyl-1,3-dioxolan-4-yl)methanol

- 38 046336 (2 mmol, 2 eq.) as a suitable alcohol, 640 mg (92%) of the target product were obtained.

Step B: methyl 5-[3-[4-[3-[tert-butoxycarbonyl(methyl)amino]prop-1-ynyl]-2fluorophenoxy]propyl]-2-[(2,2-dimethyl-1,3- dioxolan-4-yl)methylamino]thiazol-4-carboxylate.

Using the general HFIP deprotection procedure, starting with the product from Step A as the appropriate carbamate, 282 mg (51%) of the desired product was obtained.

1H NMR (400 MHz, DMSO<) δ ppm 7.73 (t, 1H), 7.31 (d, 1H), 7.21 (d, 1H), 7.13 (t, 1H), 4.23 (bs, 2H), 4.19 (q, 1H), 4.07 (t, 2H), 4.05 -3.96 (m, 2H), 3.64 (dd, 1H), 3.69 (s, 3H), 3.31-3.24 (m, 1H), 3.11 (t, 2H), 2.86 (brs, 3H), 2.05-1.95 (m , 2H), 1.41 (s, 9H), 1.33 (s, 3H), 1.26 (s, 3H).

Synthesis of 3p: methyl 5-[3-[4-[3-(dimethylamino)prop-1-ynyl]-2-fluorophenoxy]propyl]-2-[(2,2-dimethyl1,3-dioxolan-4-yl) methylamino]thiazole-4-carboxylate.

Step A: methyl 2-[tert-butoxycarbonyl-[(2,2-dimethyl-1,3-dioxolan-4-yl)methyl]amino]-5-[3-[4[3 -(dimethylamino)prop-1 -inyl]-2-fluorophenoxy]propyl]thiazole-4-carboxylate.

Using the general Mitsunobu reaction procedure, starting with 491 mg of synthesis 1c (1 mmol, 1 eq.) as the appropriate carbamate and 292 mg of (2,2-dimethyl-1,3-dioxolan-4-yl)methanol (2 mmol , 2 eq.) as a suitable alcohol, 573 mg (92%) of the desired product were obtained.

Step B: methyl 5-[3-[4-[3-(dimethylamino)prop-1-ynyl]-2-fluorophenoxy]propyl]-2-[(2,2-dimethyl1,3-dioxolan-4-yl) methylamino]thiazole-4-carboxylate.

Using the general HFIP deprotection procedure, starting with the product from Step A as the appropriate carbamate, 378 mg (74%) of synthesis product 3p was obtained.

^1H NMR (400 MHz, DMSOO δ ppm 7.74 (t, 1H), 7.30 (dd, 1H), 7.21 (d, 1H), 7.13 (t, 1H), 4.21 (qui, 1H), 4.07 ( t, 2H), 4.02-3.96 (m, 2H), 3.70 (s, 3H), 3.65 (dd, 1H), 3.42 (s, 2H), 3.33-3.26 (m, 1H), 3.12 (t, 2H) , 2.23 (s, 6H), 2.05-1.96 (m, 2H), 1.34 (s, 3H), 1.26 (s, 3H).

Synthesis of 3q: methyl 5-[3-[4-[3-[tert-butoxycarbonyl(methyl)amino]prop-1-ynyl]-2fluorophenoxy]propyl]-2-[[3-[tert-butyl(dimethyl)silyl ]oxy-2-methoxypropyl]amino]thiazole-4carboxylate.

Step A: methyl 2-[tert-butoxycarbonyl-[3-[tert-butyl(dimethyl)silyl]oxy-2methoxypropyl] amino]-5-[3-[4-[3-[tert-butoxycarbonyl(methyl)amino] prop-1-ynyl]-2fluorophenoxy]propyl]thiazole-4-carboxylate.

Using the general Mitsunobu reaction procedure, starting with 577 mg of synthesis 1b (1 mmol, 1 eq.) as the appropriate carbamate and 440 mg of synthesis 2j (2 mmol, 2 eq.) as the appropriate alcohol, yielded 780 mg (qty. .) target product.

Step B: methyl 5-[3-[4-[3-[tert-butoxycarbonyl(methyl)amino]prop-1-ynyl]-2-fluorophenoxy]propyl]-2-[[3-[tert-butyl(dimethyl )silyl]oxy-2-methoxypropyl]amino]thiazole-4-carboxylate.

Using the general HFIP deprotection procedure, starting with the product from Step A as the suitable carbamate, 400 mg (58%) of the desired product was obtained.

1H NMR (400 MHz, DMSO<) δ ppm 7.60 (t, 1H), 7.31 (d, 1H), 7.21 (d, 1H), 7.13 (t, 1H), 4.23 (bs, 2H), 4.07 (t, 2H), 3.69 (s, 3H), 3.67 -3.57 (m, 2H), 3.40-3.31 (m, 2H), 3.33 (s, 3H), 3.26-3.18 (m, 1H), 3.12 (t, 2H), 2.87 (brs, 3H), 2.01 (qui , 2H), 1.42 (s, 9H), 0.86 (s, 9H), 0.03 (s, 3H), 0.02 (s, 3H).

Synthesis of 3r: methyl 5-[3-[4-[3-[tert-butoxycarbonyl(methyl)amino]prop-1-ynyl]-2-fluorophenoxy]propyl]-2-[[3-[tert-butyl(dimethyl )silyl]oxy-2-morpholinopropyl]amino]thiazole-4-carboxylate.

Step A: methyl 2-[tert-butoxycarbonyl-[3-[tert-butyl(dimethyl)silyl]oxy-2morpholinopropyl] amino] -5 - [3-[4-[3-[tert-butoxycarbonyl(methyl)amino] prop-1-ynyl]-2fluorophenoxy]propyl]thiazole-4-carboxylate.

Using the general Mitsunobu reaction procedure, starting with 577 mg of synthesis 1b (1 mmol, 1 eq.) as the appropriate carbamate and 413 mg of synthesis 2k (1.5 mmol, 1.5 eq.) as the appropriate alcohol, gave 473 mg (56 %) of the target product.

Step B: methyl 5-[3-[4-[3-[tert-butoxycarbonyl(methyl)amino]prop-1-ynyl]-2-fluorophenoxy]propyl]-2-[[3-[tert-butyl(dimethyl )silyl]oxy-2-morpholinopropyl]amino]thiazole-4-carboxylate.

Using the general HFIP deprotection procedure, starting with the product from Step A as the suitable carbamate, 390 mg (93%) of the desired product was obtained.

1H NMR (400 MHz, DMSO<) δ ppm 7.38 (t, 1H), 7.32 (d, 1H), 7.21 (d, 1H), 7.12 (t, 1H), 4.23 (bs, 2H), 4.07 (t, 2H), 3.73 (dd, 1H), 3.69 (s, 3H), 3.64 (dd, 1H), 3.58-3.46 (m, 4H), 3.25 (t, 2H), 3.13 (t, 2H), 2.87 (brs, 3H), 2.72-2.63 (m, 3H ), 2.59-2.53 (m, 2H), 2.01 (qui, 2H), 1.42 (s, 9H), 0.86 (s, 9H), 0.03 (s, 6H).

Synthesis of 3s: methyl 5-[3-(2-fluoro-4-iodophenoxy)propyl]-2-(methylamino)thiazole-4-carboxylate.

Step A: Methyl 2-[tert-butoxycarbonyl(methyl)amino]-5-[3-(2-fluoro-4-iodophenoxy)propyl]thiazole 4-carboxylate.

Using the general alkylation procedure, starting from 2.68 g of synthesis product 1a (10 mmol, 1 eq.) and 1.70 g of iodomethane (12 mmol, 1.2 eq.) as a suitable alkyl halide, 2.8 g (77%) of the desired product were obtained.

Step B: Methyl 5-[3-(2-fluoro-4-iodophenoxy)propyl]-2-(methylamino)thiazole-4-carboxylate.

Using the general HFIP deprotection procedure based on 2.0 g of product from step A

- 39 046336 (4.44 mmol) as a suitable carbamate, 1.2 g (73%) of the target product was obtained.

1H NMR (500 MHz, DMSO-dg) δ ppm 7.59 (dd, 1H), 7.50 (q, 1H), 7.45 (m, 1H), 6.97 (t, 1H), 4.03 (t, 2H), 3.70 (s, 3H), 3.12 (t, 2H), 2.77 (d, 3H), 1.99 (m, 2H); ¹³ C NMR (125 MHz, DMSO^) δ ppm 165.4, 163.0, 147.0, 134.0, 124.9, 117.6, 68.0, 51.8, 31.1, 30.6, 23.3; MSVR-ERI (m/z): [M+H]+, calculated for C ₁₅ H ₁₇ FIN ₂ O ₃ S: 450.9983; found: 450.9971.

Synthesis 3t: methyl 2-[5-[tert-butyl(dimethyl)silyl]oxypentylamino]-5-[3-[4-[3(dimethylamino)prop-1-ynyl]-2-fluorophenoxy]propyl]thiazole-4 -carboxylate.

Using the general Sonogashira reaction procedure, starting from the crude product of synthesis 3m, step B, and KH-dimethylprop-2-yn-1-amine as the appropriate acetylene, 3.73 g (2 step amount) of the desired product was obtained.

MSHR-ERI (m/z): [M+H]+, calculated for C ₃₀ H ₄₇ FN ₃ O ₄ SSi: 592.3035; found: 592.3033.

Synthesis of 3u: ethyl 2-amino-5-(3-{4-[3-(dimethylamino)prop-1-yn-1-yl]-2-fluorophenoxy}propyl)-1,3thiazol-4-carboxylate.

Stage A: ethyl 3-bromo-6-chloro-2-oxohexanoate.

Bromine (1.6 mL, 31.2 mmol, 1.2 equiv) was added to a stirred solution of ethyl 6-chloro-2-oxohexanoate (5 g, 26.0 mmol, 1 eq.) in chloroform (70 mL). The reaction mixture was stirred at ambient temperature for 6.5 hours, then diluted with dichloromethane and washed with 10% aqueous sodium thiosulfate solution, followed by washing with water and then brine. The organic extract was dried (magnesium sulfate) and the solvent was removed in vacuo to give the title compound (7 g, 25.8 mmol, 99%) as a colorless oil.

1H NMR (400 MHz, DMSOHO δ 5.25 (dd, J=9.0, 4.7 Hz, 1H), 4.30 (q, J=7.1 Hz, 2H), 3.72 (t, J=6.3 Hz, 2H), 2.24-2.10 ( m, 1H), 2.07-1.89 (m, 2H), 1.92-1.76 (m, 1H), 1.30 (t, J=7.1 Hz, 3H).

Step B: ethyl 5-(3-chloropropyl)-2-acetamido-1,3-thiazole-4-carboxylate.

To a stirred solution of the product from step A (7 g, 25.8 mmol, 1 eq.) in ethanol (150 ml) was added N-acetylthiourea (3.05 g, 25.78 mmol, 1 eq.). The reaction mixture was refluxed for 6 hours, then allowed to cool to ambient temperature and the solvent was removed in vacuo. The residue was dissolved in dichloromethane (150 ml), then acetic anhydride (4.74 ml, 50.21 mmol, 2 eq.) and 4-dimethylaminopyridine (6.13 g, 50.21 mmol, 2 eq.) were added. After stirring at ambient temperature for 2 hours, the mixture was diluted with dichloromethane, then washed with water (x2) followed by a wash with brine. The organic extract was dried (magnesium sulfate), then the solvent was removed in vacuo. The crude material was triturated with diethyl ether and the solid was collected by filtration and washed with diethyl ether to give the title product (1.1 g, 3.78 mmol, 13.9%). Two cycles of evaporation of the filtrate, re-gritting and filtering resulted in additional batches of product (3.42 g, 11.76 g, 43%). Finally, performing evaporation of the final filtrate and purification by automated flash column chromatography (CombiFlash Rf, 12 g RediSep™ silica gel cartridge) eluting with a gradient of 0-100% ethyl acetate in isoheptane resulted in additional amounts of the desired product (1.46 g, 5.02 mmol, 18.5%) [total yield: 76%].

LC/MS (CnHi ₅ ClN ₂ O ₃ S) 291 [M+H]+; RT 1.08 (ZHMS-U-B1);

1H NMR (400 MHz, DMSOO δ 12.43 (s, 1H), 4.26 (q, J=7.1 Hz, 2H), 3.68 (t, J=6.4 Hz, 2H), 3.27 - 3.18 (m, 2H), 2.13 ( s, 3H), 2.19 - 2.00 (m, 2H), 1.29 (t, J=7.1 Hz, 3H).

Step C: ethyl 2-acetamido-5-(3-iodopropyl)-1,3-thiazole-4-carboxylate.

A solution of the product from step B (5.84 g, 20.1 mmol, 1 eq.) and sodium iodide (18.1 g, 0.12 mol, 6 eq.) in acetonitrile (130 ml) was refluxed for 6 h. The mixture was cooled to ambient temperature The medium and solvent were removed in vacuo. The mixture was diluted with ethyl acetate and washed with water and brine. The organic extract was dried (magnesium sulfate) and concentrated in vacuo. The resulting solid was triturated in diethyl ether and then collected by filtration to give the title product as a cream solid (6.28 g, 16.4 mmol, 82%).

LC/MS (C _n H ₁₅ IN ₂ O ₃ S) 383 [M+H]+; RT 0.96 (ZHMS-U-B1).

1H NMR (400 MHz, DMSOO δ 12.43 (s, 1H), 4.26 (q, J=7.1 Hz, 2H), 3.29 (t, J=6.7 Hz, 2H), 3.18 (t, J=7.4 Hz, 2H) , 2.12 (s, 3H), 2.16-2.04 (m, 2H), 1.30 (t, J=7.1 Hz, 3H).

Step D: Ethyl 5-(3-{4-[3-(dimethylamino)prop-1-yn-1-yl]-2-fluorophenoxy}propyl)-2-acetamido1,3-thiazol-4-carboxylate.

To a stirred solution of the product from step C (1.64 g, 8.46 mmol, 1.2 eq.) in dimethylformamide (20 ml) was added sodium hydride (60% in mineral oil) (0.56 g, 14.1 mmol, 2 eq.). The reaction mixture was stirred at ambient temperature for 30 min, then cooled in an ice bath and a solution of synthesis 6b (2.7 g, 7.05 mmol, 1 eq.) in dimethylformamide (20 ml) was slowly added. After 40 minutes, the reaction mixture was allowed to warm to ambient temperature and stirred for 5 hours. The solvent was removed in vacuo and the crude material was partitioned between ethyl acetate and water, washed with brine and dried (magnesium sulfate). Solvent

- 40 046336 was removed in vacuo and the crude product was purified by automated flash column chromatography (CombiFlash Rf, 80 g RediSep™ silica gel cartridge), eluting with a gradient of 0-10% methanol in dichloromethane to give the title product as a cream solid (1.48 g, 3.32 mmol, 47%).

LC/MS (C ₂₂ H ₂₆ FN ₃ O ₄ S) 448 [M+H]+; RT 0.76 (LCMS-V-BI).

¹ H NMR (400 MHz, DMSOO δ 12.41 (s, 1H), 7.30 (dd, J=11.9, 2.3 Hz, 1H), 7.25-7.18 (m, 1H), 7.177.10 (m, 1H), 4.22 ( q, J=7.1 Hz, 2H), 4.11 (t, J=6.1 Hz, 2H), 3.42 (s, 2H), 3.24 (dt, J=14.7, 7.5 Hz, 2H), 2.23 (s, 6H), 2.10 (s, 3H), 2.12 -2.01 (m, 2H), 1.27 (t, J=7.1 Hz, 3H).

Step E: ethyl 2-amino-5-(3-{4-[3-(dimethylamino)prop-1-yn-1-yl]-2-fluorophenoxy}propyl)-1,3thiazol-4-carboxylate.

A solution of the product from step D (1.49 g, 3.32 mmol, 1 eq.) and hydrochloric acid (4 M in 1,4 dioxane) (8.3 ml, 0.03 mol, 10 eq.) in ethanol (40 ml) was heated at 60°C in for 18 hours. The reaction mixture was allowed to cool to ambient temperature, then the solvent was removed in vacuo. The resulting oil was dissolved in a small amount of methanol, then loaded onto an SCX-2 cartridge (50 g, methanol washed). The cartridge was washed with methanol and then eluted with 3.5N. methanol ammonia. The solvent was removed in vacuo and the resulting oil was purified by automated flash column chromatography (CombiFlash Rf, 80 g RediSep™ silica gel cartridge), eluting with a gradient of 0-10% methanol in dichloromethane to give the title product as a cream solid (0.78 g, 1.93 mmol, 58%).

LC/MS (C ₂₀ H ₂₄ FN3O3S) 406 [M+H]+; RT 0.706 (LCMS-^B1).

¹ H NMR (400 MHz, DMSO<) δ 7.30 (dd, J= 12.0, 2.0 Hz, 1H), 7.25-7.17 (m, 1H), 7.13 (t, J=8.7 Hz, 1H), 7.02 (s, 2H), 4.16 (q, 2H), 4.07 (t, 2H), 3.42 (s, 2H), 3.12 (t, J=7.5 Hz, 2H), 2.23 (s, 6H), 2.09-1.94 (m, 2H ), 1.23 (t, J=7.1 Hz, 3H).

Synthesis 3v: ethyl 2-amino-5-(3-phenoxypropyl)-1,3-thiazole-4-carboxylate.

Step A: ethyl 5-bromo-2-acetamido-1,3-thiazole-4-carboxylate.

A solution of ethyl 2-amino-5-bromo-1,3-thiazol-4-carboxylate (2 g, 7.96 mmol, 1 eq.), acetic anhydride (0.83 ml, 8.76 mmol, 1.1 eq.) and 4-dimethylaminopyridine ( 1.12 g, 9.16 mmol, 1.15 eq.) in dichloromethane (30 ml) was stirred at ambient temperature for 3 days. The reaction mixture was diluted with dichloromethane, washed successively with water and brine, then dried (magnesium sulfate) and the solvent was removed in vacuo. The resulting solid was triturated with diethyl ether, filtered and dried in vacuo to give the title product as an off-white solid (1.8 g, 6.14 mmol, 77%).

LC/MS (C ₈ H ₉ BrN ₂ O ₃ S) 293 [M+H]+; RT 1.01 (LCMS^-B1).

¹ H NMR (400 MHz, DMSOO δ 12.80 (s, 1H), 4.28 (q, J=7.1 Hz, 2H), 2.16 (s, 3H), 1.30 (t, J=7.1 Hz, 3H).

Step B: ethyl 2-acetamido-5-(3-phenoxyprop-1-yn-1-yl)-1,3-thiazol-4-carboxylate.

Product from Step A (200 mg, 0.68 mmol, 1 eq.), phenylpropargyl ether (0.18 ml, 1.36 mmol, 2 eq.), triethylamine (0.28 ml, 2.05 mmol, 3 eq.), copper iodide ([) (13 mg, 0.07 mmol, 0.1 eq.) and tetrakis(triphenylphosphine)palladium(0) (39.4 mg, 0.03 mmol, 0.05 eq.) were combined. The mixture was bubbled with nitrogen (10 min) and then heated at 100°C for 1 h under microwave irradiation. The reaction mixture was partitioned between ethyl acetate and water, and the organics were washed with water, followed by brine, and dried (magnesium sulfate). The solvent was removed in vacuo and the crude product was purified by automated flash column chromatography (CombiFlash Rf, 12 g RediSep™ silica gel cartridge) eluting with a gradient of 0-100% ethyl acetate in isoheptane to give the title product as an off-white solid (122 mg, 0.35 mmol, 52%).

LC/MS (C ₁₇ H ₁₆ N ₂ O ₄ S) 345 [M+H]+; RT 1.21 (LCMS^-B1).

¹ H NMR (400 MHz, DMSOO δ 12.85 (s, 1H), 7.34 (t, J=7.8 Hz, 2H), 7.09-7.03 (m, 2H), 7.02-6.96 (m, 1H), 5.13 (s, 2H), 4.24 (q, J=7.0 Hz, 2H), 2.16 (s, 3H), 1.27 (t, J=7.1 Hz, 3H).

Step C: ethyl 2-acetamido-5-(3-phenoxypropyl)-1,3-thiazole-4-carboxylate.

Ethyl acetate (10 ml) was charged into a flask containing the product from step B (122 mg, 0.35 mmol, 1 eq.) and platinum oxide (8.04 mg, 0.04 mmol, 0.1 eq.) under a nitrogen atmosphere. The mixture was shaken at ambient temperature under a hydrogen atmosphere for 6.5 hours. Additional platinum(^) oxide was added (at the tip of a spatula) and the reaction mixture was shaken under a hydrogen atmosphere for an additional 18 hours. The mixture was filtered through a methanol-soaked celite pad, eluted methanol and the solvent was removed in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 12 g RediSep™ silica gel cartridge) eluting with a gradient of 0-100% ethyl acetate in isoheptane provided the title product as a cream solid (77.2 mg, 0.22 mmol, 63%).

LC/MS (C ₁₇ H ₂₀ N ₂ O ₄ S) 349 [M+H]+; RT 1.22 (LCMS^-B1).

¹ H NMR (400 MHz, DMSOO δ 12.41 (s, 1H), 7.36-7.24 (m, 2H), 7.03-6.88 (m, 3H), 4.23 (q, J=7.1 Hz, 2H), 4.00 (t, J=6.2 Hz, 2H), 3.26 (dd, J=8.3, 6.8 Hz, 2H), 2.12 (s, 3H), 2.11-2.01 (m, 2H), 1.28 (t, J=7.1 Hz, 3H).

- 41 046336

Step D: Ethyl 2-amino-5-(3-phenoxypropyl)-1,3-thiazole-4-carboxylate.

Hydrochloric acid (4 M in 1,4-dioxane; 0.55 mL, 2.22 mmol, 10 eq.) was added to a stirred solution of the product from Step C (77.2 mg, 0.22 mmol, 1 eq.) in ethanol (5 mL). The reaction mixture was heated at 60°C for 5 hours. The solvent was removed in vacuo, then the residue was dissolved in methanol and loaded onto a methanol-moistened SCX-2 cartridge (5 g). The cartridge was washed with methanol, then eluted with 3.5 N. methanolic ammonia, and the solvent was removed in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 12 g RediSep™ silica gel cartridge) eluting with a gradient of 0-10% methanol in dichloromethane gave the title product as a yellow gum (48.2 mg, 0.16 mmol, 71%).

LC/MS (C ₁₅ H ₁₈ N ₂ O ₃ S) 307 [M+H]+; RT 1.16 (ZHMS-U-B1).

1H NMR (400 MHz, DMSOO δ 7.33-7.24 (m, 2H), 7.02 (s, 2H), 6.95-6.90 (m, 3H), 4.16 (q, J=7.1 Hz, 2H), 3.97 (t, J =6.2 Hz, 2H), 3.13 (t, 2H), 2.03-1.94 (m, 2H), 1.24 (t, J=7.1 Hz, 3H).

Synthesis of 3w: ethyl 2-amino-5-(3-methoxypropyl)-1,3-thiazole-4-carboxylate.

Step A: ethyl 2-acetamido-5-(3-methoxyprop-1-yn-1-yl)-1,3-thiazol-4-carboxylate.

Dimethylformamide (10 ml) was added to the product of Synthesis 3v, step A (400 mg, 1.36 mmol, 1 eq.), methyl propargyl ether (0.23 ml, 2.73 mmol, 2 eq.), triethylamine (0.57 ml, 4.09 mmol, 3 eq. ) and copper(1) iodide (26 mg, 0.14 mmol, 0.1 eq.), and then tetrakis(triphenylphosphine)palladium(0) (78.8 mg, 0.07 mmol, 0.05 eq.) was added. The vessel was bubbled with nitrogen (10 min) and then heated at 100°C for 1 h under microwave irradiation. The solvent was removed in vacuo and the crude product was purified by automated flash column chromatography (CombiFlash Rf, 12 g RediSep™ silica gel cartridge) eluting with a gradient of 0-5% methanol in dichloromethane to give a solid, which was triturated with diethyl ether to give the title product as a cream solid (213.7 mg, 0.76 mmol, 55%). The filtrate was evaporated to obtain a second batch of the desired product as an orange gum (94.6 mg, 0.34 mmol, 24%) [79% overall yield].

LC/MS (C ₁₂ H ₁₄ N ₂ O ₄ S) 283 [M+H]+; RT 1.00 (ZHMS-U-B1).

1H NMR (400 MHz, DMSOL) δ 12.84 (s, 1H), 4.41 (s, 2H), 4.28 (q, J=7.1 Hz, 2H), 3.36 (s, 3H), 2.16 (s, 3H), 1.30 (t, J=7.1 Hz, 3H).

Step B: ethyl 2-acetamido-5-(3-methoxypropyl)-1,3-thiazole-4-carboxylate.

A solution of the product from step A (308 mg, 1.09 mmol, 1 eq.) in a 2:1 mixture of ethyl acetate/methanol (15 ml) was loaded into a flask containing a catalytic amount of platinum(IV) oxide under a nitrogen atmosphere. The vessel was evacuated and filled again with nitrogen (x3), then evacuated and placed in a hydrogen atmosphere. After shaking at ambient temperature for 18 hours, the reaction mixture was filtered through celite, eluted with methanol, and the solvent was removed in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 12 g RediSep™ silica gel cartridge) eluting with a gradient of 0-100% ethyl acetate in isoheptane provided the title product as a cream solid (207 mg, 0.72 mmol, 66%) .

LC/MS (C ₁₂ H ₁₈ N ₂ O ₄ S) 287 [M+H]+; RT 0.99 (ZHMS-U-B1).

1H NMR (400 MHz, DMSOO δ 12.40 (s, 1H), 4.25 (q, J=7.1 Hz, 2H), 3.39-3.31 (m, 2H), 3.24 (s, 3H), 3.17-3.08 (m, 2H ), 2.12 (s, 3H), 1.89-1.77 (m, 2H), 1.29 (t, J=7.1 Hz, 3H).

Step C: ethyl 2-amino-5-(3-methoxypropyl)-1,3-thiazole-4-carboxylate.

Hydrochloric acid (4 M in 1,4-dioxane; 2.1 mL, 8.38 mmol, 10 eq.) was added to a stirred solution of the product from Step B (240 mg, 0.84 mmol, 1 eq.) in ethanol (10 mL). The mixture was heated to 60°C for 7 hours, then allowed to cool to ambient temperature and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 12 g RediSep™ silica gel cartridge) eluting with a gradient of 0-10% methanol in dichloromethane gave the title product as a brown gum (219 mg, 0.81 mmol, 97%).

LC/MS (C ₁₀ H ₁₆ N ₂ O ₃ S) 245 [M+H]+; RT 0.88 (ZHMS-U-B1).

1H NMR (400 MHz, DMSOO δ 8.00 (br s, 2H), 4.25 (q, J=7.1 Hz, 2H), 3.34 (t, J=6.3 Hz, 2H), 3.23 (s, 3H), 3.05-2.96 (m, 2H), 1.84-1.72 (m, 2H), 1.28 (t, J=7.1 Hz, 3H).

Synthesis of 3x: ethyl 2-amino-5-[3-(2-fluorophenoxy)propyl]-1,3-thiazol-4-carboxylate.

Step A: ethyl 2-acetamido-5-(3-hydroxyprop-1-yn-1-yl)-1,3-thiazol-4-carboxylate.

To a mixture of synthesis product 3v, step A (1 g, 3.41 mmol, 1 eq.), propargyl alcohol (0.4 ml, 6.82 mmol, 2 eq.), triethylamine (1.42 ml, 10.2 mmol, 3 eq.) and copper iodide ( 1) (65 mg, 0.34 mmol, 0.1 eq.) in dimethylformamide (20 ml), tetrakis(triphenylphosphine) palladium(0) (197 mg, 0.17 mmol, 0.05 eq.) was added. The mixture was bubbled with nitrogen (10 min) and then heated at 100°C for 1 h under microwave irradiation. The solvent was removed in vacuo; subsequent purification by automated flash column chromatography (CombiFlash Rf, 40 g RediSep™ silica gel cartridge) eluting with a gradient of 0-10% methanol in dichloromethane gave the title product as a brown solid (1.01 g, 3.75 mmol, >100 %).

LC/MS (C11H12N2O4S) 269 [M+H]+; RT 0.82 (ZHMS-U-B1).

1H NMR (400 MHz, DMSOHO δ 12.80 (s, 1H), 5.45 (t, J=6.0 Hz, 1H), 4.37 (d, J=6.1 Hz, 2H), 4.27 (q,

- 42 046336

J=7.1 Hz, 2H), 2.16 (s, 3H), 1.30 (t, J=7.1 Hz, 3H).

Step B: ethyl 2-acetamido-5-(3-hydroxypropyl)-1,3-thiazole-4-carboxylate.

A solution of the product from step A (1.01 g, 3.75 mmol, 1 eq.) in 4:1 ethyl acetate/methanol (25 ml) was loaded into a flask containing platinum oxide hydrate ([U) (at the tip of a spatula) under a nitrogen atmosphere. The reaction mixture was evacuated and again filled with nitrogen (x3), then evacuated and placed under a hydrogen atmosphere. The reaction mixture was shaken at ambient temperature for 30 hours, then filtered through celite, eluted with methanol and the solvent was removed in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 24 g RediSep™ silica gel cartridge) eluting with a gradient of 0-10% methanol in dichloromethane gave the title product as a brown gum (582 mg, 2.14 mmol, 57%).

LC/MS (C11H16N2O4S) 273 [M+H]+; RT 0.81 (ZHMS-U-B1).

1H NMR (400 MHz, DMSO<) δ 12.39 (s, 1H), 4.54 (t, J=5.1 Hz, 1H), 4.25 (q, J=7.1 Hz, 2H), 3.44 (td, J=6.4, 5.1 Hz, 2H), 3.15-3.09 (m, 2H), 2.12 (s, 3H), 1.81-1.69 (m, 2H), 1.29 (t, J=7.1 Hz, 3H).

Step C: Ethyl 2-acetamido-5-[3-(2-fluorophenoxy)propyl]-1,3-thiazol-4-carboxylate.

A solution of 2-fluorophenol (0.1 mL, 1.1 mmol, 1.5 eq.), product from step B (200 mg, 0.73 mmol, 1 eq.) and triphenylphosphine (289 mg, 1.1 mmol, 1.5 eq.) in tetrahydrofuran (10 mL) cooled in an ice bath, then diisopropyl azobicarboxylate (0.22 mL, 1.1 mmol, 1.5 eq.) was added dropwise. The reaction mixture was allowed to gradually warm to ambient temperature and then stirred for 18 hours. The mixture was diluted with ethyl acetate, washed with brine, and the organic phase was dried (magnesium sulfate) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 24 g RediSep™ silica gel cartridge) eluting with a gradient of 0-100% ethyl acetate in heptane gave the title product as a white solid (162 mg, 0.44 mmol, 60%).

LC/MS (C ₁₇ H ₁₉ FN ₂ O ₄ S) 367 [M+H]+; RT 1.22 (ZhKhMS-U-B1).

1H NMR (400 MHz, DMSO<) δ 12.41 (s, 1H), 7.29-7.07 (m, 3H), 7.00-6.89 (m, 1H), 4.23 (q, J=7.1 Hz, 2H), 4.08 (t , J=6.2 Hz, 2H), 3.27 (dd, J=8.2, 6.9 Hz, 2H), 2.12 (s, 3H), 2.18-2.03 (m, 2H), 1.27 (t, J=7.1 Hz, 3H) .

Step D: Ethyl 2-amino-5-[3-(2-fluorophenoxy)propyl]-1,3-thiazol-4-carboxylate.

A solution of the product from step C (217 mg, 0.59 mmol, 1 eq.) and hydrochloric acid (4 M in dioxane; 1.48 mL, 5.92 mmol, 10 eq.) in ethanol (8 mL) was heated at 60°C overnight. The mixture was allowed to cool to ambient temperature, then the solvent was removed in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 12 g RediSep™ silica gel cartridge) eluting with a gradient of 0-10% methanol in dichloromethane gave the title product as a yellow foam (181 mg, 0.56 mmol, 94%).

LC/MS (C ₁₅ H ₁₇ FN ₂ O ₃ S) 325 [M+H]+; RT 1.16 (ZHMS-U-B1).

1H NMR (400 MHz, DMSO<) δ 7.62 (br s, 2H), 7.26-7.07 (m, 3H), 6.99-6.89 (m, 1H), 4.20 (q, J=7.1 Hz, 2H), 4.07 ( t, J=6.1 Hz, 2H), 3.15 (t, J=7.5 Hz, 2H), 2.08-1.99 (m, 2H), 1.25 (t, J=7.1 Hz, 3H).

Synthesis 3y: ethyl 2-amino-5-[3-(4-bromo-2-fluorophenoxy)propyl]-1,3-thiazol-4-carboxylate.

Step A: Ethyl 5-[3-(4-bromo-2-fluorophenoxy)propyl]-2-acetamido-1,3-thiazol-4-carboxylate.

Solution of synthesis product 3x, step B (248 mg, 0.91 mmol, 1 eq.), 4-bromo-2-fluorophenol (0.15 ml, 1.37 mmol, 1.5 eq.) and triphenylphosphine (358 mg, 1.37 mmol, 1.5 eq.) in tetrahydrofuran (10 ml) was cooled in an ice bath, then diisopropyl azodicarboxylate (0.27 ml, 1.37 mmol, 1.5 eq.) was added dropwise. The reaction mixture was stirred in an ice bath for 30 minutes, then stirred at ambient temperature for 3 hours. The reaction mixture was diluted with ethyl acetate and washed with water, then brine. The organics were dried (magnesium sulfate) and the solvent was removed in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 24 g RediSep™ silica gel cartridge) eluting with a gradient of 0-100% ethyl acetate in heptane provided the title product as a white solid (229 mg, 0.51 mmol, 56%) .

LC/MS (C ₁₇ H ₁₈ BrFN ₂ O ₄ S) 445 [M+H]+; RT 1.36 (ZHMS-U-B1).

1H NMR (400 MHz, DMSOO δ 12.42 (s, 1H), 7.53 (dd, J=10.9, 2.4 Hz, 1H), 7.32 (ddd, J=8.8, 2.4, 1.5 Hz, 1H), 7.13 (t, J =9.0 Hz, 1H), 4.22 (q, J=7.1 Hz, 2H), 4.08 (t, J=6.1 Hz, 2H), 3.25 (dd, J=8.4, 6.7 Hz, 2H), 2.12 (s, 3H ), 2.18-2.02 (m, 2H), 1.27 (t, J=7.1 Hz, 3H).

Step B: Ethyl 2-amino-5-[3-(4-bromo-2-fluorophenoxy)propyl]-1,3-thiazol-4-carboxylate.

A solution of the product from step A (229 mg, 0.51 mmol, 1 eq.) and hydrochloric acid (4 M in dioxane; 1.29 ml, 5.14 mmol, 10 eq.) in ethanol (8 ml) was heated at 60°C for 18 h The reaction mixture was allowed to cool to ambient temperature, then the solvent was removed in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 12 g RediSep™ silica gel cartridge) eluting with a gradient of 0-10% methanol in dichloromethane gave the title product as a yellow gum (199 mg, 0.49 mmol, 96%).

LC/MS (C ₁₅ H ₁₆ BrFN ₂ O ₃ S) 405 [M+H]+; RT 1.30 (ZHMS-U-B1).

1H NMR (400 MHz, DMSOD ₆ ) δ 7.54 (dd, J=11.0,2.4 Hz, 1H), 7.33 (ddd, J=8.8, 2.4, 1.5 Hz, 1H), 7.13 (t, J=9.0 Hz, 1H ), 4.19 (q, J=7.1 Hz, 2H), 4.07 (t, J=6.1 Hz, 2H), 3.16-3.10 (m, 2H), 2.11-1.96 (m, 2H), 1.24 (t,

- 43 046336

J=7.1 Hz, 3H).

Synthesis of 3z: ethyl 2-[(2-methoxyethyl)amino]-1,3-thiazole-4-carboxylate.

Ethyl bromopyruvate (2.5 mL, 17.9 mmol, 1.07 eq.) was added under nitrogen to a suspension of 2methoxyethylthiourea (2.25 g, 16.8 mmol, 1 eq.) in ethanol (40 mL) at 0°C, and the mixture was stirred for 1 h at environment. Triethylamine (4.7 ml, 33.5 mmol, 2 eq.) was added and the mixture was heated at 75°C for 18 hours. The solution was allowed to cool to ambient temperature and partitioned between ethyl acetate (300 ml) and water (200 ml). The organic phase was washed with water (200 ml) and brine, dried (magnesium sulfate) and concentrated in vacuo. Purification by flash column chromatography (100 g silica gel) eluting with 1:1 ethyl acetate/heptane gave a solid which was triturated with heptane (20 ml), filtered, washed with heptane (2 x 15 ml) and dried in vacuo to give the title product as a pale brown solid (3.15 g, 13.7 mmol, 82%).

LC/MS (C ₉ H ₁₄ N ₂ O ₃ S) 231 [M+H]+; RT 0.88 (LCMS-V-BI).

¹ H NMR (400 MHz, CDCl ₃ ) δ 7.40 (s, 1H), 5.78 (t, 1H), 4.34 (q, J=7.1 Hz, 2H), 3.60-3.54 (m, 2H), 3.48 (td, J=5.5, 4.4 Hz, 2H), 3.36 (s, 3H), 1.36 (t, J=7.1 Hz, 3H).

Synthesis of 3za: ethyl 5-[4-(benzyloxy)piperidin-1-yl]-2-(methylamino)-1,3-thiazol-4-carboxylate.

Step A: ethyl 5-[4-(benzyloxy)piperidin-1-yl]-1,3-thiazol-4-carboxylate.

A solution of ethyl 5-bromothiazole-4-carboxylate (500 mg, 2.12 mmol, 1 eq.), 4benzyloxypiperidine hydrochloride (579 mg, 2.54 mmol, 1.2 eq.) and 1,8-diazabicyclo[5.4.0]undec-7-ene (0.64 ml, 4.24 mmol, 2 eq.) in acetonitrile (10 ml) was heated at 100°C for 2 h under microwave irradiation. The reaction mixture was allowed to cool to ambient temperature, then concentrated in vacuo, partitioned between dichloromethane and water, and the organic phase was washed with brine, dried (magnesium sulfate) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 24 g RediSep™ silica gel cartridge) eluting with a gradient of 0-100% ethyl acetate in isoheptane gave the title product as a yellow oil (499 mg, 1.44 mmol, 68%).

LC/MS (C ₁₈ H ₂₂ N ₂ O ₃ S) 347 [M+H]+; RT 1.08 (LCMS-V-BI).

1H NMR (400 MHz, DMSO^) δ 8.46 (s, 1H), 7.41-7.33 (m, 4H), 7.32-7.26 (m, 1H), 4.55 (s, 2H), 4.25 (q, J=7.1 Hz , 2H), 3.62 (tt, J=7.7, 3.6 Hz, 1H), 3.42-3.30 (m, 2H), 3.08-2.97 (m, 2H), 2.05-1.95 (m, 2H), 1.81-1.66 (m , 2H), 1.28 (t, J=7.1 Hz, 3H).

Step B: ethyl 5-[4-(benzyloxy)piperidin-1-yl]-2-bromo-1,3-thiazol-4-carboxylate.

N-Bromosuccinimide (308 mg, 1.73 mmol, 1.2 eq.) was added to a stirred solution of the product from Step A (499 mg, 1.44 mmol, 1 eq.) in acetonitrile (20 ml), and the mixture was stirred at ambient temperature overnight . The reaction mixture was diluted with ethyl acetate, washed with water followed by brine, dried (magnesium sulfate) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 24 g RediSep™ silica gel cartridge) eluting with a gradient of 0-60% ethyl acetate in isoheptane provided the title product as a yellow solid (454 mg, 1.07 mmol, 74%) .

LC/MS (C ₁₈ H ₂₁ BrN ₂ O ₃ S) 427 [M+H]+; RT 1.26 (LCMS-V-BI).

1H NMR (400 MHz, DMSO<) δ 7.41-7.33 (m, 4H), 7.32-7.26 (m, 1H), 4.54 (s, 2H), 4.24 (q, J=7.1 Hz, 2H), 3.62 (tt , J=7.5, 3.5 Hz, 1H), 3.43-3.28 (m, 2H), 3.07 (ddd, J=11.8, 8.5, 3.4 Hz, 2H), 2.06-1.88 (m, 2H), 1.79-1.65 (m , 2H), 1.28 (t, J=7.1 Hz, 3H).

Step C: ethyl 5-[4-(benzyloxy)piperidin-1-yl]-2-{[(tert-butoxy)carbonyl](methyl)amino}-1,3thiazol-4-carboxylate.

The product from step B (454 mg, 1.07 mmol, 1 eq.) and tert-butyl methyl carbamate (0.19 ml, 1.28 mmol, 1.2 eq.) were added under a nitrogen atmosphere to a solution of tris(dibenzylideneacetone)dipalladium(0) (48.9 mg, 0.05 mmol, 0.05 eq.) and xantphos (61.8 mg, 0.11 mmol, 0.1 eq.) in 1,4-dioxane (10 ml). Cesium carbonate (522 mg, 1.6 mmol, 1.5 eq) was added and the mixture was heated at 100°C overnight. The reaction mixture was allowed to cool to ambient temperature and concentrated in vacuo. The residue was dissolved in ethyl acetate, then washed with water followed by brine, dried (magnesium sulfate) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 50 g RediSep™ silica gel cartridge) eluting with a gradient of 0-50% ethyl acetate in isoheptane provided the title product as a cream solid (339 mg, 0.71 mmol, 67%).

LC/MS (C ₂₄ H ₃₃ N ₃ O5S) 476 [M+H]+; RT 1.37 (LCMS-V-BI).

1H NMR (400 MHz, DMSO<) δ 7.41-7.33 (m, 4H), 7.32-7.26 (m, 1H), 4.55 (s, 2H), 4.23 (q, J=7.1 Hz, 2H), 3.59 (tt , J=8.0, 3.7 Hz, 1H), 3.38 (s, 3H), 3.37-3.29 (m, 2H), 2.97 (ddd, J=11.6, 8.8, 3.1 Hz, 2H), 2.06 - 1.91 (m, 2H ), 1.76-1.62 (m, 2H), 1.52 (s, 9H), 1.28 (t, J=7.1 Hz, 3H).

Step D: Ethyl 5-[4-(benzyloxy)piperidin-1-yl]-2-(methylamino)-1,3-thiazol-4-carboxylate.

Trifluoroacetic acid (0.53 ml, 7.11 mmol, 10 eq.) at 0 °C was added to a stirred solution of the product from step C (339 mg, 0.71 mmol, 1 eq.) in dichloromethane (10 ml), and the mixture was allowed to

- 44 046336 warm to ambient temperature and stir overnight. The reaction mixture was partitioned between dichloromethane and saturated aqueous sodium bicarbonate, and the organic phase was washed with brine, dried (magnesium sulfate) and concentrated in vacuo. The material was triturated with diethyl ether, filtered and dried in vacuo to give the title product as a white solid (213 mg, 0.57 mmol, 80%).

LC/MS (C ₁₉ H ₂₅ N ₃ O ₃ S) 376 [M+H]+; RT 1.07 (LCMS-V-BI).

1H NMR (400 MHz, DMSOZH) δ 7.38-7.33 (m, 3H), 7.32-7.23 (m, 2H), 4.53 (s, 2H), 4.18 (q, J=7.1 Hz, 2H), 3.54 (dt, J=8.2, 4.3 Hz, 1H), 3.19-3.08 (m, 2H), 2.83-2.72 (m, 5H), 2.02-1.89 (m, 2H), 1.76-1.60 (m, 2H), 1.26 (t, J=7.1 Hz, 3H).

Synthesis of 3zb: ethyl 5-{1-[(tert-butoxy)carbonyl]azetidin-3-yl}-2-(methylamino)-1,3-thiazol-4carboxylate.

Step A: ({1-[(tert-butoxy)carbonyl]azetidin-3-yl}methyl)(iodo)zinc.

An oven-dried Schlenk flask was charged with zinc (3.3 g, 50.5 mmol, 1.5 equiv.), and the flask was heated at 160°C for 20 min in vacuo, then allowed to cool to ambient temperature and created a nitrogen atmosphere. Dimethylacetamide (25 mL) was added followed by slow addition of a mixture of trimethylchlorosilane (0.69 mL, 5.42 mmol, 0.16 eq.) and 1,2-dibromoethane (0.49 mL, 5.69 mmol, 0.17 eq.). The resulting mixture was allowed to cool to ambient temperature, then a solution of 1-boc-3-(iodomethyl)azetidine (10 g, 33.65 mmol, 1 eq.) in dimethylacetamide (7 ml) was slowly added and the mixture was stirred overnight. Cannulation through a filter (cotton wool/celite/cotton wool) into a dry 25 ml Schlenk flask resulted in the target product as a 0.62 M solution (as determined by titration with 0.5 M iodine solution), which was used without further characterization.

Step B: tert-butyl 3-(3-ethoxy-2,3-dioxopropyl)azetidine-1-carboxylate.

A solution of copper iodide® (3.54 g, 18.6 mmol, 2 eq.) in tetrahydrofuran (20 ml) was cooled to -20°C, then the product from step A (0.62 M in dimethylacetamide; 30 ml, 18.6 mmol, 2 eq.) was slowly added. ), and the mixture was stirred at -20°C for 10 minutes and then at 0°C for 20 minutes. The mixture was cooled to 20°C, then ethyl chloroglyoxylate (1.04 mL, 9.3 mmol, 1 eq.) was added slowly and the mixture was allowed to warm to ambient temperature and stirred for 1 hour. The reaction mixture was quenched by adding saturated aqueous ammonium chloride, extracted with ethyl acetate (x2) and the combined organic extracts were washed with brine, dried (magnesium sulfate) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 40 g RediSep™ silica gel cartridge) eluting with a gradient of 0-30% ethyl acetate in isoheptane gave the title product as a colorless oil (0.57 g, 2.1 mmol, 23%).

1H NMR (400 MHz, Chloroform-d) δ 4.33 (q, J=7.2 Hz, 2H), 4.13 (t, J=8.5 Hz, 2H), 3.57 (dd, J=8.9, 5.4 Hz, 2H), 3.20 (d, J=7.5 Hz, 2H), 2.99-2.84 (m, 1H), 1.43 (s, 9H), 1.37 (t, J=7.1 Hz, 3H).

Step C: tert-butyl 3-[(12)-3-ethoxy-3-oxo-2-[(trimethylsilyl)oxy]prop-1-en-1-yl]azetidine-1carboxylate.

Triethylamine (0.38 mL, 2.73 mmol, 1.3 eq) was slowly added to a solution of the product from step B (570 mg, 2.1 mmol, 1 eq.) and chlorotrimethylsilane (0.3 mL, 2.35 mmol, 1.12 eq.) in tetrahydrofuran (10 mL). , and the mixture was stirred at ambient temperature for 3.5 hours. The reaction mixture was partitioned between ethyl acetate and water, and the organic phase was washed with brine, dried (magnesium sulfate) and concentrated in vacuo to give the title product as a colorless oil (643 mg, 1.87 mmol, 89%).

1H NMR (400 MHz, Chloroform-d) δ 6.23 (d, J=8.9 Hz, 1H), 4.22 (q, J=7.1 Hz, 2H), 4.14 (t, J=8.5 Hz, 2H), 3.74 (dd , J=8.4, 6.0 Hz, 2H), 3.61-3.46 (m, 1H), 1.44 (s, 9H), 1.32 (t, J=7.1 Hz, 3H), 0.20 (s, 9H).

Step D: tert-butyl 3-(1-bromo-3-ethoxy-2,3-dioxopropyl)azetidine-1-carboxylate.

A solution of the product from step C (643 mg, 1.87 mmol, 1 eq.) and N-bromosuccinimide (367 mg, 2.06 mmol, 1.1 eq.) in tetrahydrofuran (15 ml) was refluxed for 2.5 h and then allowed to cool to ambient temperature. The reaction mixture was partitioned between ethyl acetate and saturated aqueous sodium bicarbonate, and the organic phase was washed with brine, dried (magnesium sulfate) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 24 g RediSep™ silica gel cartridge) eluting with a gradient of 0-40% ethyl acetate in isoheptane gave the title product as a yellow gum (507 mg, 1.45 mmol, 77%).

1H NMR (400 MHz, Chloroform-d) δ 5.25 (d, J=9.3 Hz, 1H), 4.40 (q, 2H), 4.14 (t, 1H), 4.08 (t, J=8.8 Hz, 1H), 3.85 (dd, J=9.1, 5.5 Hz, 1H), 3.58 (dd, J=9.3, 5.2 Hz, 1H), 3.25-3.10 (m, 1H), 1.44 (s, 9H), 1.40 (t, J=7.2 Hz, 3H).

Step E: ethyl 5-(azetidin-3-yl)-2-(methylamino)-1,3-thiazol-4-carboxylate.

A solution of the product from step D (452 mg, 1.29 mmol, 1 eq.) and N-methylthiourea (140 mg, 1.55 mmol, 1.2 eq.) in ethanol (8 ml) was refluxed for 4.5 h and then allowed to cool to ambient temperature. The resulting precipitate was collected by filtering

- 45 046336 trituration, washed with ethanol (2 ml), and dried in vacuum to obtain the title product as a white solid (161 mg, 0.5 mmol, 39%). The filtrate was transferred to a microwave vessel and hydrochloric acid (4 M in 1,4-dioxane, 0.2 ml, 0.8 mmol) was added and the mixture was heated at 60°C for 40 min under microwave irradiation. The reaction mixture was loaded onto a methanol-conditioned SCX-2 cartridge (10 g), then washed with methanol, and eluted with 3.5 N. methanol ammonia. The solvent was removed in vacuo and the crude product was purified using automated flash column chromatography (CombiFlash Rf, 24 g RediSep™ silica gel cartridge) eluting with a 0-15% gradient of 2.33 N. methanolic ammonia in dichloromethane to obtain the target product in the form of a colorless resin (115 mg, 0.48 mmol, 37%). The two batches of product were combined and used in the following steps.

LC/MS (C ₁₀ H ₁₅ N ₃ O2S) 242 [M+H]+; RT 0.34 (LCMS-V-BI).

¹ H NMR (400 MHz, DMSOO δ 8.84 (s, 1H), 8.57 (s, 1H), 7.83 (q, J=4.8 Hz, 1H), 4.76 (p, J=8.8 Hz, 1H), 4.32-4.17 (m, 4H), 3.95 (q, J=8.0 Hz, 2H), 2.82 (d, J=4.7 Hz, 3H), 1.28 (t, J=7.1 Hz, 3H).

Step F: ethyl 2-{[(tert-butoxy)carbonyl](methyl)amino}-5-{1-[(tert-butoxy)carbonyl]azetidin3 -yl}-1,3-thiazol-4-carboxylate.

To a solution of the product from step E (298 mg, 1.23 mmol, 1 eq.) in dichloromethane (10 ml) was added ditert-butyl dicarbonate (809 mg, 3.7 mmol, 3 eq.), 4-(dimethylamino)pyridine (45.3 mg, 0.37 mmol, 0.3 eq.) and triethylamine (0.51 ml, 3.7 mmol, 3 eq.), and the mixture was stirred at ambient temperature overnight. The reaction mixture was partitioned between dichloromethane and water, and the organic phase was washed with brine, dried (magnesium sulfate) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 24 g RediSep™ silica gel cartridge) eluting with a gradient of 0-40% ethyl acetate in isoheptane gave the title product as a white foam (390 mg, 0.88 mmol, 72%).

LC/MS (C ₂₀ H ₃₁ N ₃ O ₆ S) 442 [M+H]+; RT 1.47 (LCMS-V-BI).

1H NMR (400 MHz, DMSOO δ 4.53 (tt, J=8.6, 5.8 Hz, 1H), 4.38-4.30 (m, 2H), 4.27 (q, 4H), 3.85-3.76 (m, 2H), 3.45 (s , 3H), 1.54 (s, 9H), 1.40 (s, 9H), 1.30 (t, J=7.1 Hz, 3H).

Step G: ethyl 5-{1-[(tert-butoxy)carbonyl]azetidin-3-yl}-2-(methylamino)-1,3-thiazol-4carboxylate.

A solution of the product from step F (390 mg, 0.88 mmol, 1 eq.) in 1,1,1,3,3,3-hexafluoro-2-propanol (3 ml) was heated at 100°C for 55 min under microwave irradiation . The mixture was concentrated in vacuo and purified by automated flash column chromatography (CombiFlash Rf, 12 g RediSep™ silica gel cartridge) eluting with a gradient of 0-100% ethyl acetate in isoheptane to give the title product as a white solid (199 mg, 0.58 mmol , 66%).

LC/MS (C15H23N3O4S) 342 [M+H]+; RT 0.98 (LCMS-V-BI).

1H NMR (400 MHz, DMSOO δ 7.70 (q, J=4.7 Hz, 1H), 4.49 (tt, J=8.6, 5.9 Hz, 1H), 4.28 (t, J=8.6 Hz, 2H), 4.21 (q, J=7.1 Hz, 2H), 3.73-3.65 (m, 2H), 2.81 (d, J=4.7 Hz, 3H), 1.39 (s, 9H), 1.26 (t, J=7.1 Hz, 3H).

Synthesis of 3zc: methyl 5-(3-{4-[3-(dimethylamino)prop-1-yn-1-yl]-2-fluorophenoxy}-2,2-dimethylpropyl)2-(methylamino)-1,3 - thiazole-4-carboxylate.

Step A: 4-methoxy-2,2-dimethyl-4-oxobutanoic acid.

To a solution of 2,2-dimethylsuccinic acid (20 g, 137 mmol, 1 eq.) in methanol (200 ml) cooled to 0°C, sulfuric acid (2 ml) was added and the mixture was stirred at ambient temperature overnight . The reaction mixture was concentrated in vacuo, then quenched by adding saturated aqueous sodium bicarbonate (200 ml) in portions, and washed with isoheptane (x2). The aqueous phase was cooled to 0°C, acidified with 6 N. aqueous solution of hydrochloric acid, and extracted with diethyl ether (x2). The combined organic extracts were dried (magnesium sulfate) and concentrated in vacuo to give the title product as a clear oil (13.7 g, 85.4 mmol, 62%).

1H NMR (400 MHz, Chloroform-d) δ 3.67 (s, 3H), 2.62 (s, 2H), 1.30 (s, 6H).

Step B: Methyl 4-hydroxy-3,3-dimethylbutanoate.

A solution of the product from step A (13.7 g, 85.4 mmol, 1 eq.) in tetrahydrofuran (140 ml) was cooled to -30°C, and the borane-dimethyl sulfide complex (13 g, 16.2 ml, 171 mmol, 2 eq.) was added dropwise. ). The mixture was stirred at -30°C for 1.5 hours, then heated to 0°C and stirred at this temperature for 45 minutes. The reaction mixture was quenched by slow addition of methanol (50 ml) and then concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 220 g RediSep™ silica gel cartridge) eluting with a gradient of 0-50% ethyl acetate in isoheptane gave the title product as a clear oil (8.14 g, 55.7 mmol, 65%).

1H NMR (400 MHz, DMSO^) δ 4.65 (t, J=5.4 Hz, 1H), 3.57 (s, 3H), 3.15 (d, J=5.4 Hz, 2H), 2.19 (s, 2H), 0.89 ( s, 6H).

Step C: Methyl 4-[(tert-butyldiphenylsilyl)oxy]-3,3-dimethylbutanoate.

To a solution of the product from step B (8.14 g, 55.7 mmol, 1 eq.) in dimethylformamide (200 ml) add

- 46 046336 tert-butyl(chloro)diphenylsilane (17.4 ml, 66.8 mmol, 1.2 eq.) and imidazole (7.36 ml, 111 mmol, 2 eq.), and the mixture was stirred at ambient temperature overnight. The reaction mixture was partitioned between heptane (500 ml) and 2:1 water/saturated aqueous ammonium chloride (500 ml) and the aqueous phase was extracted with heptane (500 ml). The combined organic extracts were dried (magnesium sulfate) and purified by automated flash column chromatography (CombiFlash Rf, 330 g RediSep™ silica gel cartridge) eluting with a gradient of 0-10% ethyl acetate in isoheptane to give the title product as a clear oil (14.4 g , 37.4 mmol, 67%).

1H NMR (400 MHz, DMSO^) δ 7.65-7.58 (m, 4H), 7.53-7.40 (m, 6H), 3.56 (s, 3H), 3.41 (s, 2H), 2.33 (s, 2H), 1.01 (s, 9H), 0.97 (s, 6H).

Step D: 4-[(tert-butyldiphenylsilyl)oxy]-3,3-dimethylbutan-1-ol.

A solution of the product from step C (14.4 g, 37.4 mmol, 1 eq.) in tetrahydrofuran (70 ml) was cooled to 78°C and added diisobutylaluminum hydride (1 M in tetrahydrofuran; 112 ml, 112 mmol, 3 eq.), and the mixture stirred at -78°C for 2 hours. A saturated aqueous ammonium chloride solution was added and the mixture was stirred at -78°C for 10 minutes and then allowed to warm to ambient temperature. The mixture was filtered through a pad of celite, eluting with ethyl acetate, and the organics were dried (magnesium sulfate) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 220 g RediSep™ silica gel cartridge) eluting with a gradient of 0-15% ethyl acetate in isoheptane gave the title product as a clear oil (12.5 g, 35 mmol, 94%).

LC/MS (C ₂₂ H ₃₂ O ₂ Si) 357 [M+H]+; RT 2.68 (LCMS-VC).

1H NMR (400 MHz, DMSO^) δ 7.65-7.56 (m, 4H), 7.53-7.40 (m, 6H), 4.28 (t, J=5.0 Hz, 1H), 3.50-3.41 (m, 2H), 3.30 (s, 2H), 1.48 (dd, J=8.1, 7.1 Hz, 2H), 1.02 (s, 9H), 0.88 (s, 6H).

Step E: 4-[(tert-butyldiphenylsilyl)oxy]-3,3-dimethylbutyl methanesulfonate.

To a stirred solution of the product from step D (12.5 g, 35 mmol, 1 eq.) in dichloromethane (200 ml), cooled to 0°C, was added triethylamine (7.31 ml, 52.5 mmol, 1.5 eq.) and methanesulfonyl chloride (3.25 ml, 42 mmol, 1.2 eq.), and the mixture was stirred at ambient temperature overnight. The reaction mixture was diluted with dichloromethane, washed with saturated aqueous sodium bicarbonate, dried (PTFE separator) and concentrated in vacuo to give the title product as a yellow oil (15.1 g, 34.8 mmol, 99%).

1H NMR (400 MHz, DMSOHO δ 7.66-7.58 (m, 4H), 7.53-7.40 (m, 6H), 4.27 (t, J=7.4 Hz, 2H), 3.33 (s, 2H), 3.15 (s, 3H ), 1.74 (t, J=7.4 Hz, 2H), 1.02 (s, 9H), 0.92 (s, 6H).

Step F: (4-bromo-2,2-dimethylbutoxy)(tert-butyl)diphenylsilane.

To a solution of the product from step E (15.1 g, 34.8 mmol, 1 eq.) in tetrahydrofuran (400 ml), lithium bromide (9.07 g, 105 mmol, 3 eq.) was added in portions and the mixture was stirred at 70°C for 4 h , and then allowed to cool to ambient temperature. The reaction mixture was concentrated in vacuo, then partitioned between dichloromethane and brine, and the organic phase was washed with 10% aqueous sodium thiosulfate, dried (PTFE phase separator) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 220 g RediSep™ silica gel cartridge) eluting with a gradient of 0-2% ethyl acetate in isoheptane gave the title product as a clear oil (12.8 g, 30.5 mmol, 88%).

1H NMR (400 MHz, DMSOHO δ 7.65-7.55 (m, 4H), 7.53-7.40 (m, 6H), 3.52-3.42 (m, 2H), 3.34 (s, 2H), 1.96-1.86 (m, 2H) , 1.03 (s, 9H), 0.88 (s, 6H).

Step G: Ethyl 6-[(tert-butyldiphenylsilyl)oxy]-5,5-dimethyl-2-oxohexanoate.

The oven-dried flask was charged with magnesium (496 mg, 20.4 mmol, 1.2 eq) and the solid was stirred vigorously under nitrogen for 20 min. A solution of the product from step F (7.13 g, 17 mmol, 1 eq.) in tetrahydrofuran (12 ml) was bubbled with nitrogen (10 min) and an aliquot of this solution was added to the magnesium (without stirring). The mixture was briefly heated under reflux (heat dryer) and an iodine crystal was added. Stirring was started and the remaining solution was added to the magnesium suspension at a rate that allowed gentle reflux to be maintained. After addition was complete, the mixture was refluxed for 2 hours, then allowed to cool to ambient temperature. A stirred solution of diethyl oxalate (2.31 mL, 17 mmol, 1 eq.) in tetrahydrofuran (6 mL) was bubbled with nitrogen (10 min) and then cooled to 78°C. The above-mentioned Grignard reagent solution was added to the solution in portions and the mixture was then allowed to gradually warm to ambient temperature over 2 hours. The reaction mixture was partitioned between ethyl acetate and saturated aqueous ammonium chloride, and the organic phase was dried (magnesium sulfate) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 220 g RediSep™ silica gel cartridge) eluting with a gradient of 0-8% ethyl acetate in isoheptane gave the title product as a clear oil (3.06 g, 6.95 mmol, 41%).

1H NMR (400 MHz, Chloroform-d) δ 7.67-7.61 (m, 4H), 7.45-7.34 (m, 6H), 4.31 (q, J=7.0 Hz, 2H), 3.31 (s, 2H), 2.80- 2.71 (m, 2H), 1.67-1.58 (m, 2H), 1.36 (s, 3H), 1.05 (s, 9H), 0.88 (s, 6H).

- 47 046336

Step GH: ethyl 3-bromo-6-[(tert-butyldiphenylsilyl)oxy]-5,5-dimethyl-2-oxohexanoate.

To a stirred mixture of copper(II) bromide (3.1 g, 13.9 mmol, 2 eq.) in ethyl acetate (80 ml), a solution of the product from step G (3.06 g, 6.94 mmol, 1 eq.) in chloroform (40 ml) was added in portions. and the reaction mixture was refluxed for 16 hours. The reaction mixture was allowed to cool to ambient temperature, then filtered through a pad of celite, eluted with dichloromethane, and evaporated. Purification by automated flash column chromatography (CombiFlash Rf, 80 g RediSep™ silica gel cartridge) eluting with a gradient of 0-10% ethyl acetate in isoheptane gave the title product as a clear oil (2.62 g, 5.04 mmol, 72%).

LC/MS (C ₂₆ H ₃₅ BrO ₄ Si) 441 [M-Ph] ⁺ ; RT 1.36 (ZHMS-U-V2).

¹ H NMR (400 MHz, Chloroform-d) δ 7.69-7.61 (m, 4H), 7.50-7.34 (m, 6H), 5.28 (dd, J=8.4, 4.5 Hz, 1H), 4.36 (qd, J= 7.1, 1.4 Hz, 2H), 3.33 (d, J=1.5 Hz, 2H), 2.56 (dd, J=14.9, 8.5 Hz, 1H), 2.11 (dd, J=14.9, 4.5 Hz, 1H), 1.38 ( t, J=7.1 Hz, 3H), 1.07 (s, 9H), 0.88 (d, J=3.7 Hz, 6H).

Step H: ethyl 5-{3-[(tert-butyldiphenylsilyl)oxy]-2,2-dimethylpropyl}-2-(methylamino)-1,3thiazole-4-carboxylate.

To a solution of the product from step GH (2.62 g, 5.04 mmol, 1 eq.) in ethanol (35 ml) was added Nmethylthiourea (455 mg, 5.04 mmol, 1 eq.) and triethylamine (1.4 ml, 10.1 mmol, 2 eq.), and the mixture was heated at 85°C for 4 hours, and then allowed to cool to ambient temperature. The reaction mixture was partitioned between dichloromethane and brine, and the organic phase was dried (PTFE phase separator) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 80 g RediSep™ silica gel cartridge) eluting with a gradient of 0-50% ethyl acetate in isoheptane gave the title product as a yellow solid (1.45 g, 2.84 mmol, 56%) .

LC/MS (C ₂₈ H ₃₈ N ₂ O ₃ SiS) 511 [M+H] ⁺ ; RT 1.28 (ZHMS-U-V2).

1H NMR (400 MHz, DMSOO δ 7.66-7.58 (m, 4H), 7.52-7.40 (m, 6H), 4.15 (q, J=7.1 Hz, 2H), 3.37 (s, 2H), 3.08 (s, 2H ), 2.77 (d, J=4.8 Hz, 3H), 1.20 (t, 3H), 1.04 (s, 9H), 0.86 (s, 6H).

Step I: Ethyl 2-{[(tert-butoxy)carbonyl](methyl)amino}-5-{3-[(tert-butyldiphenylsilyl)oxy]2,2-dimethylpropyl}-1,3-thiazole-4-carboxylate .

To a solution of the product from step H (1 g, 1.96 mmol, 1 eq.) in tetrahydrofuran (20 ml) was added 4(dimethylamino)pyridine (23.9 mg, 0.2 mmol, 0.1 eq.), triethylamine (0.82 ml, 5.87 mmol, 3 eq.) and ditert-butyl dicarbonate (0.84 ml, 3.92 mmol, 2 eq.), and the mixture was stirred at ambient temperature for 3 hours. The reaction mixture was partitioned between ethyl acetate and water, and the organic phase was dried (magnesium sulfate) and concentrated in a vacuum. Purification by automated flash column chromatography (CombiFlash Rf, 24 g RediSep™ silica gel cartridge) eluting with a gradient of 0-15% ethyl acetate in isoheptane gave the title product as a yellow oil (1.19 g, 1.95 mmol, 100%).

LC/MS (C ₃₃ H ₄₆ N ₂ O ₅ SiS) 611 [M+H] ⁺ ; RT 1.48 (ZHMS-U-V2).

1H NMR (400 MHz, DMSOO δ 7.68-7.60 (m, 4H), 7.52-7.39 (m, 6H), 4.22 (q, J=7.1 Hz, 2H), 3.44 (s, 3H), 3.37 (s, 2H ), 3.20 (s, 2H), 1.54 (s, 9H), 1.23 (t, 3H), 1.04 (s, 9H), 0.87 (s, 6H).

Step J: 2-{[(tert-butoxy)carbonyl](methyl)amino}-5-(3-hydroxy-2,2-dimethylpropyl)-1,3-thiazole4-carboxylic acid.

To a solution of the product from Step I (600 mg, 0.98 mmol, 1 eq.) in tetrahydrofuran (12 ml) was added TBAF (1 M in tetrahydrofuran; 2.95 ml, 2.95 mmol, 3 eq.) and the mixture was stirred at ambient temperature at for 2 hours. The reaction mixture was partitioned between ethyl acetate and water, and the organic phase was dried (magnesium sulfate) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 12 g RediSep™ silica gel cartridge) eluting with a gradient of 0-100% ethyl acetate in isoheptane provided the title product as a white solid (178 mg, 0.52 mmol, 53%) .

LC/MS (C ₁₅ H ₂₄ N ₂ O ₅ S) 345 [M+H] ⁺ ; RT 1.87 (ZHMS-U-S).

1H NMR (400 MHz, DMSOO δ 12.69 (br s, 1H), 4.72 (br s, 1H), 3.34 (s, 3H), 3.16 (s, 2H), 3.10 (s, 2H), 1.53 (s, 9H ), 0.83 (s, 6H).

Step K: methyl 2-{[(tert-butoxy)carbonyl](methyl)amino}-5-(3-hydroxy-2,2-dimethylpropyl)-1,3thiazole-4-carboxylate.

To a solution of the product from step J (178 mg, 0.52 mmol, 1 eq.) in a 1:1 mixture of dichloromethane/methanol (4 ml), cooled to 0°C, was added (trimethylsilyl)diazomethane (2M in hexanes; 0.31 ml, 0.62 mmol, 1.2 eq.), and the mixture was stirred at ambient temperature overnight. The reaction mixture was partitioned between dichloromethane and water, and the organic phase was dried (PTFE phase separator) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 4 g RediSep™ silica gel cartridge) eluting with a gradient of 050% ethyl acetate in isoheptane provided the title product as a white solid (119 mg, 0.33 mmol, 64%).

LC/MS (C16H26N2O5S) 359 [M+H] ⁺ ; RT 2.10 (ZHMS-U-S).

- 48 046336 ¹ H NMR (400 MHz, DMSO-dg) δ 4.74 (t, J=5.1 Hz, 1H), 3.78 (s, 3H), 3.43 (s, 3H), 3.16 (d, J=5.2 Hz, 2H), 3.10 (s, 2H), 1.53 (s, 9H), 0.82 (s, 6H).

Step L: methyl 2-{[(tert-butoxy)carbonyl](methyl)amino}-5-[3-(2-fluoro-4-iodophenoxy)-2,2dimethylpropyl]-1,3-thiazol-4-carboxylate .

To a solution of the product from step K (119 mg, 0.33 mmol, 1 eq.) in toluene (2 ml) was added 2-fluoro-4iodophenol (158 mg, 0.66 mmol, 2 eq.), triphenylphosphine (174 mg, 0.66 mmol, 2 eq.) and diisopropyl azodicarboxylate (131 μl, 0.66 mmol, 2 eq.), and the mixture was heated at 120°C for 24 h. The reaction mixture was partitioned between dichloromethane and water, and the organic phase was dried (PTFE phase separator) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 4 g RediSep™ silica gel cartridge) eluting with a gradient of 020% ethyl acetate in isoheptane gave the title product as a clear gum (136 mg, 0.24 mmol, 71%).

LC/MS (C ₂₂ H ₂₈ FIN ₂ O ₅ S) 579 [M+H]+; RT 2.76 (LCMS-VC).

1H NMR (400 MHz, DMSO<) δ 7.60 (dd, J=10.7, 2.1 Hz, 1H), 7.49 - 7.42 (m, 1H), 3.75 (s, 5H), 3.42 (s, 3H), 3.26 (s , 2H), 1.49 (s, 9H), 1.01 (s, 6H).

Step M: methyl 2-{[(tert-butoxy)carbonyl](methyl)amino}-5-(3-{4-[3-(dimethylamino)prop-1-yn1-yl]-2-fluorophenoxy}-2 ,2-dimethylpropyl)-1,3-thiazole-4-carboxylate.

To a solution of the product from step L (136 mg, 0.24 mmol, 1 eq.) in tetrahydrofuran (6 ml) was added 3-dimethylamino-1-propyne (38.1 μl, 0.35 mmol, 1.5 eq.), copper iodide ([) (4.48 mg, 0.02 mmol, 0.1 eq.) and N.N-diisopropylethylamine (81.9 μl, 0.47 mmol, 2 eq.). The mixture was bubbled with nitrogen (10 min), then bis(triphenylphosphine)palladium(II) dichloride (16.5 mg, 0.02 mmol, 0.1 eq.) was added and the mixture was heated at 70°C overnight. The reaction mixture was partitioned between ethyl acetate and brine, and the organic phase was dried (magnesium sulfate) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 4 g RediSep™ silica gel cartridge) eluting with a gradient of 0-4% methanol in dichloromethane gave the title product as a brown gum (27 mg, 0.05 mmol, 22%).

LC/MS (C ₂₇ H3 ₆ FN ₃ O5S) 534 [M+H]+; RT 2.053 (LCMS-VC).

1H NMR (400 MHz, DMSOD ₆ ) δ 7.31 (dd, J=11.9, 2.0 Hz, 1H), 7.25-7.18 (m, 1H), 7.13 (t, J=8.7 Hz, 1H), 3.78 (s, 2H ), 3.75 (s, 3H), 3.43 (s, 2H), 3.42 (s, 3H), 3.27 (s, 2H), 2.24 (s, 6H), (s, 9H), 1.02 (s, 6H).

Step N: methyl 5-(3-{4-[3-(dimethylamino)prop-1-yn-1-yl]-2-fluorophenoxy}-2,2-dimethylpropyl)-2(methylamino)-1,3- thiazole-4-carboxylate.

To a solution of the product from step M (27 mg, 0.05 mmol, 1 eq.) in dichloromethane (5 ml) was added trifluoroacetic acid (1 ml) and the mixture was stirred at ambient temperature for 2 hours. The reaction mixture was diluted with dichloromethane, then cooled to 0°C and neutralized by adding 2N. aqueous sodium hydroxide solution. The layers were separated and dried (PTFE phase separator), and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 4 g RediSep™ silica gel cartridge) eluting with a gradient of 0-7% methanol in dichloromethane provided the title product as a brown solid (8 mg, 0.02 mmol, 37%) .

LC/MS (C ₂₂ H ₂₈ FN ₃ O ₃ S) 434 [M+H]+; RT 1.62 (LCMS-VC).

1H NMR (400 MHz, DMSOO δ 7.52-7.46 (m, 1H), 7.32 (dd, J=11.8, 2.0 Hz, 1H), 7.24-7.19 (m, 1H), 7.13 (t, J=8.7 Hz, 1H ), 3.76 (s, 2H), 3.71 (s, 3H), 3.42 (s, 2H), 3.16 (s, 2H), 2.76 (d, J=4.8 Hz, 3H), 2.23 (s, 6H), 1.00 (s, 6H).

Synthesis of 3zd: methyl 5-{[1-({4-[3-(dimethylamino)prop-1-in-1-yl]-2-fluorophenoxy}methyl)cyclopropyl]methyl}-2-(methylamino)-1, 3-thiazole-4-carboxylate.

Step A: 2-(1-{ [(tert-butyldiphenylsilyl)oxy]methyl}cyclopropyl)acetonitrile.

To a solution of 2-[1-(hydroxymethyl)cyclopropyl]acetonitrile (8 g, 72 mmol, 1 eq.) in dimethylformamide (200 ml) was added tert-butyl(chloro)diphenylsilane (22.5 ml, 86.4 mmol, 1.2 eq.) and imidazole (9.51 ml, 144 mmol, 2 eq.) and the mixture was stirred at ambient temperature overnight. The reaction mixture was partitioned between isoheptane (600 ml) and 2:1 water/saturated aqueous ammonium chloride (600 ml) and the aqueous phase was extracted with isoheptane (600 ml). The combined organic extracts were dried (magnesium sulfate) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 330 g RediSep™ silica gel cartridge) eluting with a gradient of 0-6% ethyl acetate in isoheptane gave the title product as a clear oil (23 g, 65.7 mmol, 91%).

1H NMR (400 MHz, DMSOD ₆ ) δ 7.67-7.57 (m, 4H), 7.53-7.36 (m, 6H), 3.54 (s, 2H), 2.71 (s, 2H), 1.02 (s, 9H), 0.57 -0.50 (m, 2H), 0.50-0.43 (m, 2H).

Step B: 2-(1-{[(tert-butyldiphenylsilyl)oxy]methyl}cyclopropyl)acetaldehyde.

To a solution of the product from step A (23 g, 65.7 mmol, 1 eq.) in dichloromethane (300 ml), cooled to 0°C, diisobutylaluminum hydride (1 M in hexanes; 72.3 ml, 72.3 mmol, 1.1 eq.) was added in portions. , and the mixture was stirred at 0°C for 1.5 h. The reaction mixture was quenched by adding water, then

- 49 046336 added 2n. an aqueous solution of hydrochloric acid (100 ml) followed by the addition of a saturated aqueous solution of potassium sodium tartrate (500 ml). The mixture was extracted with dichloromethane (2x400 ml) and the combined organic extracts were dried (magnesium sulfate) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 330 g RediSep™ silica gel cartridge) eluting with a gradient of 0-10% ethyl acetate in isoheptane gave the title product as a clear oil (19.6 g, 55.5 mmol, 85%).

LC/MS (C ₂₂ H ₂₈ O ₂ Si) 353 [M+H]+; RT 1.19 (ZHMS-U-V2).

1H NMR (400 MHz, DMSO-dg) δ 9.77 (t, J=2.5 Hz, 1H), 7.61-7.56 (m, 4H), 7.50-7.38 (m, 6H), 3.33 (s, 2H), 2.41 ( d, J=2.5 Hz, 2H), 0.97 (s, 9H), 0.57-0.50 (m, 2H), 0.47-0.40 (m, 2H).

Step C: 2-(1-{ [(tert-butyldiphenylsilyl)oxy]methyl}cyclopropyl)ethan-1-ol.

To a solution of the product from step B (19.6 g, 55.5 mmol, 1 eq.) in methanol (200 ml), cooled to 0°C, sodium borohydride (2.31 g, 61.1 mmol, 1.1 eq.) was added in portions and the mixture was stirred at ambient temperature for 3 hours. The reaction mixture was concentrated in vacuo, then partitioned between ethyl acetate and water, and the organic phase was dried (magnesium sulfate) and concentrated in vacuo to give the title product as a clear oil (18.8 g, 53 mmol, 96% ).

1H NMR (400 MHz, DMSO-de) δ 7.66-7.57 (m, 4H), 7.52-7.37 (m, 6H), 4.32 (t, J=5.1 Hz, 1H), 3.57-3.47 (m, 2H), 3.46 (s, 2H), 1.59 (t, J=7.3 Hz, 2H), 1.00 (s, 9H), 0.35-0.26 (m, 4H).

Step P: 2-(1-{[(tert-butyldiphenylsilyl)oxy]methyl}cyclopropyl)ethyl methanesulfonate.

To a solution of the product from step C (18.8 g, 53 mmol, 1 eq.) in dichloromethane (300 ml), cooled to 0°C, was added triethylamine (22.1 ml, 159 mmol, 3 eq.) and methanesulfonyl chloride (4.92 ml, 63.6 mmol, 1.2 eq.), and the mixture was stirred at ambient temperature overnight. The reaction mixture was partitioned between dichloromethane and saturated aqueous sodium bicarbonate, and the organic phase was separated and dried (PTFE phase separator), and concentrated in vacuo to give the desired product as a brown oil (23.3 g, 53 mmol, 100%).

1H NMR (400 MHz, DMSOO δ 7.67-7.56 (m, 4H), 7.53-7.37 (m, 6H), 4.31 (t, J=7.1 Hz, 2H), 3.48 (s, 2H), 3.13 (s, 3H ), 1.84 (t, J=7.1 Hz, 2H), 1.01 (s, 9H), 0.49-0.38 (m, 2H), 0.36-0.29 (m, 2H).

Step E: {[1-(2-bromoethyl)cyclopropyl]methoxy}(tert-butyl)diphenylsilane.

To a solution of the product from step D (23.3 g, 53.9 mmol, 1 eq) in tetrahydrofuran (600 ml) was added lithium bromide (14 g, 162 mmol, 3 eq) and the mixture was heated at 70°C for 5 h, then allowed to cool to ambient temperature and the solvent was removed in vacuo. The residue was partitioned between dichloromethane and brine, and the organic phase was washed with 10% aqueous sodium thiosulfate, dried (magnesium sulfate) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 330 g RediSep™ silica gel cartridge) eluting with a gradient of 0-2% diethyl ether in isoheptane gave the title product as a clear oil (17.4 g, 41.6 mmol, 77%) .

1H NMR (400 MHz, DMSO-de) δ 7.65-7.58 (m, 4H), 7.53-7.39 (m, 6H), 3.54 (dd, J=8.6, 7.4 Hz, 2H), 3.47 (s, 2H), 1.98 (dd, J=8.6, 7.3 Hz, 2H), 1.02 (s, 9H), 0.46-0.38 (m, 2H), 0.35-0.28 (m, 2H).

Step F: Ethyl 4-(1-{[(tert-butyldiphenylsilyl)oxy]methyl}cyclopropyl)-2-oxobutanoate.

The oven-dried flask was charged with magnesium (1.21 g, 49.9 mmol, 1.2 eq) and the solid was stirred vigorously under nitrogen for 20 min. A solution of the product from step E (17.4 g, 41.6 mmol, 1 eq.) in tetrahydrofuran (30 ml) was bubbled with nitrogen (10 min), then an aliquot of this solution was added to the magnesium (without stirring), the mixture was briefly refluxed and the crystal was added Yoda. Stirring was started and the remaining solution was added to the magnesium at a rate that allowed gentle reflux to be maintained. After addition was complete, the mixture was refluxed for 2 hours, then allowed to cool to ambient temperature. A solution of diethyl oxalate (5.64 mL, 41.6 mmol, 1 eq.) in tetrahydrofuran (15 mL) was bubbled with nitrogen (10 min) and then cooled to -78°C. The above Grignard reagent solution was added slowly and the mixture was then allowed to warm to ambient temperature over 2 hours. The reaction mixture was partitioned between ethyl acetate and saturated aqueous ammonium chloride, and the organic phase was dried (magnesium sulfate) and concentrated in vacuo.

Purification by automated flash column chromatography (CombiFlash Rf, 330 g RediSep™ silica gel cartridge) eluting with a gradient of 0-8% ethyl acetate in isoheptane gave the title product as a clear oil (10 g, 22.8 mmol, 55%).

1H NMR (400 MHz, DMSO-de) δ 7.65-7.57 (m, 4H), 7.53-7.36 (m, 6H), 4.19 (q, J=24.7, 7.1 Hz, 2H), 3.45 (s, 2H), 2.89-2.80 (m, 2H), 1.67-1.59 (m, 2H), 1.33-1.17 (m, 5H), 1.05-0.96 (s, 11H).

Step G: Ethyl 3-bromo-4-(1-{[(tert-butyldiphenylsilyl)oxy]methyl}cyclopropyl)-2-oxobutanoate.

A solution of the product from step F (10 g, 22.8 mmol, 1 eq.) in chloroform (125 ml) was added to copper(II) bromide (10.2 g, 45.6 mmol, 2 eq.) in ethyl acetate (250 ml) and the mixture was heated at 85°C overnight. The reaction mixture was filtered through celite, eluted with dichloromethane and concentrated in vacuo. Purification using automated flash column chromatography (CombiFlash Rf, 220 g RediSep™ silica gel cartridge) eluting with a gradient of 0-8% ethyl acetate in isoheptane resulted in

- 50 046336 to obtain the target product in the form of a yellow oil (2.09 g, 4.04 mmol, 18%).

1H NMR (400 MHz, DMSO-dg) δ 7.68-7.59 (m, 4H), 7.53-7.40 (m, 6H), 5.52 (dd, J=8.9, 5.2 Hz, 1H), 4.28 (q, 2H), 3.68 (d, 1H), 3.37 (s, 1H), 2.44 (dd, J=15.2, 5.2 Hz, 1H), 2.02-1.88 (m, 1H), 1.26 (t, 3H), 1.03 (s, 9H) , 0.62-0.53 (m, 1H), 0.48-0.39 (m, 1H), 0.39-0.32 (m, 1H), 0.32-0.23 (m, 1H).

Step H: ethyl 5-[(1-{[(tert-butyldiphenylsilyl)oxy]methyl}cyclopropyl)methyl]-2-(methylamino)1,3-thiazole-4-carboxylate.

To a solution of the product from step G (2.09 g, 4.04 mmol, 1 eq.) in ethanol (30 ml) was added Nmethylthiourea (364 mg, 4.04 mmol, 1 eq.) and triethylamine (1.12 ml, 8.08 mmol, 2 eq.), and the mixture was heated at 85°C overnight. The reaction mixture was partitioned between dichloromethane and brine, and the organic phase was dried (PTFE phase separator) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 40 g RediSep™ silica gel cartridge) eluting with a gradient of 0-50% ethyl acetate in isoheptane gave the title product as an orange solid (1.8 g, 3.53 mmol, 88%) .

LC/MS (C41 h.NCLSiS) 509 [M+H]+; RT 2.69 (LCMS-V-C).

1H NMR (400 MHz, DMSOO δ 7.59-7.51 (m, 4H), 7.46-7.36 (m, 6H), 4.16 (q, J=7.1 Hz, 2H), 3.44 (s, 2H), 3.22 (s, 2H ), 2.79 (d, J=4.8 Hz, 3H), 1.20 (t, 3H), 0.96 (s, 9H), 0.55-0.48 (m, 2H), 0.48-0.41 (m, 2H).

Step I: ethyl 2-{[(tert-butoxy)carbonyl](methyl)amino}-5-[(1-{[(tert-butyldiphenylsilyl)oxy]methyl} cyclopropyl)methyl]-1,3-thiazol-4 -carboxylate.

To a solution of the product from step H (1.8 g, 3.54 mmol, 1 eq.) in tetrahydrofuran (40 ml) was added 4(dimethylamino)pyridine (43.2 mg, 0.35 mmol, 0.1 eq.), triethylamine (1.48 ml, 10.6 mmol, 3 eq.) and di-tert-butyl dicarbonate (1.51 ml, 7.08 mmol, 2 eq.), and the mixture was stirred at ambient temperature overnight. The reaction mixture was partitioned between ethyl acetate and water, and the organic phase was dried (magnesium sulfate) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 40 g RediSep™ silica gel cartridge) eluting with a gradient of 0-15% ethyl acetate in isoheptane gave the title product as a yellow oil (2.03 g, 3.33 mmol, 94%).

LC/MS (C ₃₃ H ₄₄ N ₂ O ₅ SiS) 609 [M+H]+; RT 1.98 (LCMS-VC).

1H NMR (400 MHz, DMSOO δ 7.56-7.48 (m, 4H), 7.46-7.34 (m, 6H), 4.23 (q, J=7.1 Hz, 2H), 3.48-3.42 (m, 5H), 3.32 (s , 2H), 1.53 (s, 9H), 1.24 (t, 3H), 0.94 (s, 9H), 0.62-0.55 (m, 1H), 0.55-0.46 (m, 1H).

Step J: 2-{[(tert-butoxy)carbonyl](methyl)amino}-5-{[1-(hydroxymethyl)cyclopropyl]methyl}1,3-thiazole-4-carboxylic acid.

To a solution of the product from Step I (2.03 g, 3.33 mmol, 1 eq.) in tetrahydrofuran (35 ml) was added tetrabutylammonium fluoride (1 M in tetrahydrofuran; 10 ml, 10 mmol, 3 eq.) and the mixture was stirred at 70°C for 2.5 hours. The reaction mixture was partitioned between ethyl acetate and water, and the organic phase was dried (magnesium sulfate) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 24 g RediSep™ silica gel cartridge) eluting with a gradient of 0-100% ethyl acetate in isoheptane provided the title product as a yellow solid (460 mg, 1.34 mmol, 40%) .

LC/MS (C ₁₅ H ₂₂ N ₂ O ₅ S) 343 [M+H]+; RT 1.77 (LCMS-VC).

1H NMR (400 MHz, DMSOO δ 4.54 (br s, 1H), 3.44 (s, 3H), 3.22 (s, 2H), 3.19 (s, 2H), 1.53 (s, 9H), 0.50-0.42 (m, 4H).

Step K: methyl 2-{[(tert-butoxy)carbonyl](methyl)amino}-5-{[1-(hydroxymethyl)cyclopropyl]methyl}-1,3-thiazol-4-carboxylate.

To a solution of the product from step J (460 mg, 1.34 mmol, 1 eq.) in a 1:1 mixture of dichloromethane/methanol (12 ml), cooled to 0°C, was added (trimethylsilyl)diazomethane (2 M in hexanes; 0.87 ml 1.75 mmol, 1.3 eq.), and the mixture was stirred at ambient temperature overnight. The reaction mixture was partitioned between dichloromethane and water, and the organic phase was dried (PTFE phase separator) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 4 g RediSep™ silica gel cartridge) eluting with a gradient of 0-50% ethyl acetate in isoheptane provided the title product as a white solid (418 mg, 1.17 mmol, 87%) .

LC/MS (C ₁₆ H ₂₄ N ₂ O ₅ S) 357 [M+H]+; RT 1.98 (LCMS-VC).

1H NMR (400 MHz, DMSOO δ 4.54 (t, J=5.5 Hz, 1H), 3.78 (s, 3H), 3.44 (s, 3H), 3.22 (d, 2H), 3.20 (s, 2H), 1.53 ( s, 9H), 0.51-0.44 (m, 4H).

Step L: methyl 2-{[(tert-butoxy)carbonyl](methyl)amino}-5-({1-[(2-fluoro-4iodophenoxy)methyl]cyclopropyl}methyl)-1,3-thiazol-4- carboxylate.

To a solution of the product from step K (418 mg, 1.17 mmol, 1 eq.) in toluene (10 ml) was added 2-fluoro-4iodophenol (558 mg, 2.35 mmol, 2 eq.), triphenylphosphine (615 mg, 2.35 mmol, 2 eq.) and diisopropyl azodicarboxylate (0.46 ml, 2.35 mmol, 2 eq.), and the mixture was heated at 120°C overnight, then allowed to cool to ambient temperature. The reaction mixture was partitioned between dichloromethane and water, and the organic phase was separated and dried (PTFE phase separator), and concentrated in vacuo.

- 51 046336

Purification by automated flash column chromatography (CombiFlash Rf, 12 g RediSep™ silica gel cartridge) eluting with a gradient of 0-20% ethyl acetate in isoheptane gave the title product as a yellow oil (649 mg, 1.13 mmol, 96%).

LC/MS (C ₂₂ H ₂₆ FIN ₂ O ₅ S) 577 [M+H]+; RT 2.65 (LCMS-VC).

1H NMR (400 MHz, DMSO-db) δ 7.56 (dd, J=10.8, 2.1 Hz, 1H), 7.39 (dt, J=8.6, 1.7 Hz, 1H), 6.84 (t, J=8.8 Hz, 1H) , 3.75 (s, 3H), 3.42 (s, 3H), 3.34 (s, 2H), 3.31 (s, 2H), 1.52 (s, 9H), 0.77-0.64 (m, 4H).

Step M: methyl 2-{[(tert-butoxy)carbonyl](methyl)amino}-5-{[1-({4-[3-(dimethylamino)prop-1in-1-yl]-2-fluorophenoxy} methyl)cyclopropyl]methyl}-1,3-thiazole-4-carboxylate.

To a solution of the product from step L (649 mg, 1.13 mmol, 1 eq.) in tetrahydrofuran (12 ml) was added 3-dimethylamino-1-propyne (182 μl, 1.69 mmol, 1.5 eq.), copper iodide ([) (21.4 mg, 0.11 mmol, 0.1 eq.) and N.N-diisopropylethylamine (392 μl, 2.25 mmol, 2 eq.). The mixture was bubbled with nitrogen (10 min), then bis(triphenylphosphine)palladium(II) dichloride (79 mg, 0.11 mmol, 0.1 eq.) was added and the mixture was heated at 80°C for 24 h. The reaction mixture was partitioned between ethyl acetate and hydrochloric acid. solution and the organic phase was dried (magnesium sulfate) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 24 g RediSep™ silica gel cartridge) eluting with a gradient of 0-7% methanol in dichloromethane provided the title product as a brown solid (173 mg, 0.33 mmol, 29%) .

LC/MS (C ₂₇ H3 ₄ FN ₃ O5S) 532 [M+H]+; RT 2.004 (LCMS-VC).

1H NMR (400 MHz, DMSO-de) δ 7.27 (dd, J=11.9, 2.0 Hz, 1H), 7.18-7.10 (m, 1H), 6.99 (t, J=8.7 Hz, 1H), 3.79 (s, 2H), 3.65 (s, 3H), 3.42 (s, 5H), 3.32 (s, 2H), 2.23 (s, 6H), 1.52 (s, 9H), 0.77-0.65 (m, 4H).

Step N: methyl 5-{[1-({4-[3-(dimethylamino)prop-1-in-1-yl]-2-fluorophenoxy}methyl)cyclopropyl]methyl}-2-(methylamino)-1, 3-thiazole-4-carboxylate.

To a solution of the product from step M (173 mg, 0.33 mmol, 1 eq.) in dichloromethane (10 ml) was added trifluoroacetic acid (2 ml) and the mixture was stirred at ambient temperature for 2 hours. The reaction mixture was diluted with dichloromethane, then cooled to 0°C and neutralized by adding 2N. aqueous sodium hydroxide solution. The organic phase was separated and dried (PTFE phase separator), and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 4 g RediSep™ silica gel cartridge) eluting with a gradient of 07% methanol in dichloromethane provided the title product as a yellow solid (84 mg, 0.19 mmol, 60%).

LC/MS (C ₂₂ H ₂₆ FN ₃ O ₃ S) 432 [M+H]+; RT 1.55 (LCMS-VC).

1H NMR (400 MHz, DMSO-de) δ 7.51 (q, J=4.7 Hz, 1H), 7.28 (dd, J=11.9, 2.0 Hz, 1H), 7.19-7.12 (m, 1H), 7.01 (t, J=8.7 Hz, 1H), 3.78 (s, 2H), 3.60 (s, 3H), 3.42 (s, 2H), 3.20 (s, 2H), 2.78 (d, J=4.7 Hz, 3H), 2.23 ( s, 6H), 0.69-0.60 (m, 4H).

Synthesis of 3ze: ethyl 2-{[4-(tert-butoxy)-4-oxobutyl]amino}-1,3-thiazole-4-carboxylate.

Step A: ethyl 2-{[4-(tert-butoxy)-4-oxobutyl][(tert-butoxy)carbonyl]amino}-1,3-thiazole-4carboxylate.

To a solution of ethyl 2-[(tert-butoxycarbonyl)amino]-1,3-thiazol-4-carboxylate (5.2 g, 19.1 mmol, 1 eq.) and tert-butyl 4-hydroxybutanoate (4.59 g, 28.6 mmol, 1.5 eq. .) in tetrahydrofuran (150 ml), triphenylphosphine (7.51 g, 28.6 mmol, 1.5 eq.) was added, followed by dropwise addition of diisopropyl azodicarboxylate (5.64 ml, 28.6 mmol, 1.5 eq.), and the mixture was stirred at ambient temperature for 1 h The reaction mixture was partitioned between ethyl acetate and water, the aqueous phase was extracted with ethyl acetate (3x60 ml) and the combined organic extracts were washed with brine (100 ml), dried (magnesium sulfate) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 120 g RediSep™ silica gel cartridge) eluting with a gradient of 0-20% ethyl acetate in isoheptane gave the title product as a colorless oil (7.7 g, 18.6 mmol, 97%).

LC/MS (Ci ₉ H ₃₀ N ₂ O ₆ S) 415 [M+H]+; RT 1.49 (ZHMS-U-B1).

1H NMR (400 MHz, DMSOO δ 8.07 (s, 1H), 4.27 (q, J=7.1 Hz, 2H), 4.12 -4.02 (m, 2H), 2.26 (t, J=7.2 Hz, 2H), 1.90 ( p, J=7.1 Hz, 2H), 1.54 (s, 9H), 1.37 (s, 9H), 1.30 (t, J=7.1 Hz, 3H).

Step B: ethyl 2-{[4-(tert-butoxy)-4-oxobutyl]amino}-1,3-thiazole-4-carboxylate.

A solution of the product from step A (7.7 g, 18.6 mmol, 1 eq.) in 1,1,1,3,3,3-hexafluoro-2-propanol (40 ml) was heated in a stoppered flask at 80°C for 7 hours The reaction mixture was concentrated in vacuo and purified by automated flash column chromatography (CombiFlash Rf, 80 g RediSep™ silica gel cartridge) eluting with a gradient of 0-35% ethyl acetate in isoheptane to give the title product as a white solid (3.61 g, 11.5 mmol, 62%).

LC/MS (C ₁₄ H ₂₂ N ₂ O ₄ S) 315 [M+H]+; RT 1.20 (ZHMS-U-B1).

1H NMR (400 MHz, DMSOO δ 7.82 (t, J=5.4 Hz, 1H), 7.50 (s, 1H), 4.21 (q, J=7.1 Hz, 2H), 3.22 (td, J=7.0, 5.4 Hz, 2H), 2.29 (t, J=7.4 Hz, 2H), 1.76 (p, J=7.2 Hz, 2H), 1.40 (s, 9H), 1.26 (t, J=7.1 Hz, 3H).

Synthesis of 3zf: ethyl 5-[(2S)-3-{4-[3-(dimethylamino)prop-1-in-1-yl]-2-fluorophenoxy}-2-methylpropyl]2-(methylamino)-1, 3-thiazole-4-carboxylate.

- 52 046336

Step A: [(2R)-3-bromo-2-methylnpronoxy](tert-butyl)diphenylsilane.

Imidazole (4.32 ml, 65.4 mmol, 2 eq.) and tert-butyl(chloro)diphenylsilane (11.1 ml, 42.5 mmol, 1.3 eq.), and the mixture was stirred at ambient temperature overnight. The reaction mixture was partitioned between isoheptane (300 ml) and 2:1 water/saturated aqueous ammonium chloride (200 ml), the aqueous phase was extracted with isoheptane (200 ml) and the combined organic extracts were dried (magnesium sulfate) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 220 g RediSep™ silica gel cartridge) eluting with a gradient of 0-2% diethyl ether in isoheptane gave the title product as a clear oil (12.1 g, 30.9 mmol, 95%) .

Ή NMR (400 MHz, DMSOO δ 7.67-7.59 (m, 4H), 7.53-7.40 (m, 6H), 3.63 (d, 2H), 3.61-3.54 (m, 2H), 2.10-1.98 (m, 1H) , 1.01 (s, 9H), 0.96 (d, 3H).

Step B: (3S)-4-[(tert-butyldiphenylsilyl)oxy]-3-methylbutanenitrile.

To a solution of the product from step A (12.1 g, 30.9 mmol, 1 eq) in dimethyl sulfoxide (120 ml) was added sodium cyanide (4.54 g, 92.7 mmol, 3 eq) and the mixture was heated at 120°C for 5 h. The reaction mixture was partitioned between diethyl ether and water, and the organic phase was dried (magnesium sulfate) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 220 g RediSep™ silica gel cartridge) eluting with a gradient of 0-3% ethyl acetate in isoheptane gave the title product as a clear oil (7.17 g, 21.2 mmol, 69%).

Ή NMR (400 MHz, DMSO^ ₆ ) δ 7.67-7.57 (m, 4H), 7.55-7.40 (m, 6H), 3.62-3.46 (m, 2H), 2.68-2.53 (m, 2H), 2.12-1.96 (m, 1H), 1.02 (m, 9H), 0.98 (d, 3H).

Step C: ^)-4-[(tert-butyldiphenylsilyl)oxy]-3-methylbutanal.

To a solution of the product from step B (7.17 g, 21.2 mmol, 1 eq.) in dichloromethane (100 ml) cooled to 0°C, diisobutylaluminum hydride (23.4 ml, 23.4 mmol, 1.1 eq.) was added and the mixture was stirred for nights. Water was added followed by 2N. an aqueous solution of hydrochloric acid (36 ml) and a saturated aqueous solution of potassium sodium tartrate (180 ml). The mixture was diluted with additional water and dichloromethane, and the organic phase was washed successively with water and brine, dried (PTFE phase separator) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 220 g RediSep™ silica gel cartridge) eluting with a gradient of 0-6% ethyl acetate in isoheptane gave the title product as a clear oil (6.21 g, 18.2 mmol, 86%).

Ή NMR (400 MHz, DMSOL) δ 9.70 (t, J=1.9 Hz, 1H), 7.66-7.57 (m, 4H), 7.53-7.40 (m, 6H), 3.56-3.42 (m, 2H), 2.57- 2.44 (m, 1H), 2.37-2.20 (m, 2H), 0.99 (s, 9H), 0.90 (d, J=6.0 Hz, 3H).

Step D: (3S)-4-[(tert-butyldiphenylsilyl)oxy]-3-methylbutan-1-ol.

To a solution of the product from step C (6.21 g, 18.24 mmol, 1 eq.) in methanol (80 mL) cooled in an ice water bath, sodium borohydride (759 mg, 20.1 mmol, 1.1 eq.) was added and the mixture was allowed to warm to ambient temperature and stirred for 3 hours. The reaction mixture was concentrated in vacuo, then partitioned between ethyl acetate and water, and the organic phase was dried (magnesium sulfate) and concentrated in vacuo to give the title product as a clear oil (6.36 g, 18.6 mmol , > 100%).

Ή NMR (400 MHz, DMSOL) δ 7.66-7.56 (m, 4H), 7.52-7.39 (m, 6H), 4.35 (t, J=5.1 Hz, 1H), 3.56-3.34 (m, 4H), 1.85- 1.70 (m, 1H), 1.65-1.51 (m, 1H), 1.31-1.21 (m, 1H), 1.01 (s, 9H), 0.91 (d, J=6.7 Hz, 3H).

Step E: (3S)-4-[(tert-butyldiphenylsilyl)oxy]-3-methylbutyl methanesulfonate.

To a cooled solution of the product from step D (6.36 g, 18.6 mmol, 1 eq.) in dichloromethane (100 ml) was added triethylamine (7.75 ml, 55.7 mmol, 3 eq.) and methanesulfonyl chloride (1.72 ml, 22.3 mmol, 1.2 eq.) , and the mixture was stirred at ambient temperature overnight. The reaction mixture was partitioned between dichloromethane and saturated aqueous sodium bicarbonate, and the organic phase was separated and dried (PTFE phase separator), and concentrated in vacuo to give the desired product as a brown oil (8.28 g, 19.7 mmol, >100%).

Ή NMR (400 MHz, DMSO^6) δ 7.66-7.58 (m, 4H), 7.53-7.40 (m, 6H), 4.25 (t, J=6.6 Hz, 2H), 3.56-3.46 (m, 2H), 3.15 (s, 3H), 1.95-1.76 (m, 2H), 1.59-1.46 (m, 1H), 1.01 (s, 9H), 0.93 (d, J=6.7 Hz, 3H).

Step F: [(2S)-4-bromo-2-methylbutoxy](tert-butyl)diphenylsilane.

To a solution of the product from step E (8.28 g, 19.7 mmol, 1 eq.) in tetrahydrofuran (180 ml) was added lithium bromide (5.13 g, 59.1 mmol, 3 eq.) and the mixture was heated at 70°C for 5 h. The reaction mixture was concentrated in vacuo, the residue was partitioned between dichloromethane and brine, and the organic phase was separated and dried (PTFE phase separator), and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 120 g RediSep™ silica gel cartridge) eluting with a gradient of 0-2% diethyl ether in isoheptane gave the title product as a clear oil (6.3 g, 15.5 mmol, 79%) .

Ή NMR (400 MHz, DMSO^ ₆ ) δ 7.66-7.58 (m, 4H), 7.53-7.40 (m, 6H), 3.64-3.47 (m, 4H), 2.08-1.93 (m,

- 53 046336

1H), 1.92-1.79 (m, 1H), 1.74-1.60 (m, 1H), 1.01 (s, 9H), 0.90 (d, J=6.8 Hz, 3H).

Step G: ethyl (5S)-6-[(tert-butyldiphenylsilyl)oxy]-5-methyl-2-oxohexanoate.

Magnesium (453 mg, 18.7 mmol, 1.2 eq.) was charged into an oven-dried flask under nitrogen atmosphere and the solid was stirred vigorously for 20 minutes, and then the stirring was stopped. A solution of the product from step F (6.3 g, 15.5 mmol, 1 eq.) in tetrahydrofuran (12 ml) was prepared and an aliquot was added to magnesium followed by iodine (39.4 mg, 0.16 mmol, 0.01 eq.) and the mixture was briefly heated with reflux condenser. The remaining solution was added at a rate that allowed gentle reflux to be maintained, and after addition was complete, the mixture was heated for 2 hours at reflux and then allowed to cool to ambient temperature. To a stirred solution of diethyl oxalate (2.11 mL, 15.5 mmol, 1 eq.) in tetrahydrofuran (6 mL), cooled to -78°C, the Grignard reagent solution was added via cannula and the mixture was allowed to warm to ambient temperature and stirred for 5 h The reaction mixture was partitioned between ethyl acetate and saturated aqueous ammonium chloride, and the organic phase was dried (magnesium sulfate) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 80 g RediSep™ silica gel cartridge) eluting with a gradient of 0-8% ethyl acetate in isoheptane gave the title product as a clear oil (4.13 g, 9.68 mmol, 62%).

1H NMR (400 MHz, DMSO^ ₆ ) δ 7.65-7.57 (m, 4H), 7.53-7.38 (m, 6H), 4.32-4.18 (m, 2H), 3.53-3.45 (m, 2H), 2.86-2.74 (m, 1H), 1.74-1.61 (m, 1H), 1.32-1.14 (m, 6H), 1.01 (s, 9H), 0.88 (d, 3H).

Step H: ethyl (5S)-3-bromo-6-[(tert-butyldiphenylsilyl)oxy]-5-methyl-2-oxohexanoate.

To a mixture of copper(II) bromide (4.14 g, 18.5 mmol, 2 eq.) in ethyl acetate (120 ml) was added a solution of the product from step G (3.95 g, 9.26 mmol, 1 eq.) in chloroform (60 ml), and the mixture heated at 85°C overnight. The reaction mixture was filtered through celite, eluting with dichloromethane, then concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 80 g RediSep™ silica gel cartridge) eluting with a gradient of 0-8% ethyl acetate in isoheptane gave the title product as a yellow oil (1.92 g, 3.8 mmol, 41%), which was directly used in the next step without characterization.

Step I: ethyl 5-[(2S)-3-[(tert-butyldiphenylsilyl)oxy]-2-methylpropyl]-2-(methylamino)-1,3thiazole-4-carboxylate.

To a solution of the product from step H (1.67 g, 3.3 mmol, 1 eq.) in ethanol (30 ml) was added Nmethylthiourea (298 mg, 3.3 mmol, 1 eq.) followed by the addition of triethylamine (0.92 ml, 6.61 mmol, 2 eq. ), and the mixture was heated at 80°C for 4 hours. The reaction mixture was allowed to cool to ambient temperature and the solvent was removed by rotary evaporation. The crude material was partitioned between dichloromethane and water, the aqueous phase was extracted with dichloromethane, and the combined organics were washed with brine, dried (magnesium sulfate) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 40 g RediSep™ silica gel cartridge) eluting with a gradient of 0-50% ethyl acetate in isoheptane gave the title product as a brown gum (914 mg, 1.84 mmol, 56%).

LC/MS (C ₂₇ H ₃₆ N ₂ O ₃ SiS) 497 [M+H]+; RT 1.46 (ZHMS-U-B1).

1H NMR (400 MHz, DMSOO δ 7.64-7.56 (m, 4H), 7.55-7.36 (m, 6H), 4.16 (q, J=7.1 Hz, 2H), 3.60-3.47 (m, 2H), 3.08 (dd , J=14.4, 6.6 Hz, 1H), 2.92 (dd, J=14.4, 7.7 Hz, 1H), 2.78 (d, J=4.8 Hz, 3H), 1.96-1.84 (m, 1H), 1.21 (t, J=7.1 Hz, 3H), 1.01 (s, 9H), 0.92 (d, J=6.7 Hz, 3H).

Step J: ethyl 2-{[(tert-butoxy)carbonyl](methyl)amino}-5-[(2S)-3-[(tert-butyldiphenylsilyl)oxy]-2-methylpropyl]-1,3-thiazol- 4-carboxylate.

To a solution of the product from stage I (914 mg, 1.84 mmol, 1 eq.) in tetrahydrofuran (20 ml) was added di-tert-butyl dicarbonate (803 mg, 3.68 mmol, 2 eq.) followed by the addition of triethylamine (0.77 ml, 5.52 mmol, 3 eq.) and 4-(dimethylamino)pyridine (22.5 mg, 0.18 mmol, 0.1 eq.), and the mixture was stirred at ambient temperature for 2 hours. The reaction mixture was partitioned between ethyl acetate and water, and the organic phase was washed with hydrochloric acid solution, dried (magnesium sulfate) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 24 g RediSep™ silica gel cartridge) eluting with a gradient of 0-20% ethyl acetate in isoheptane gave the title product as a brown gum (961 mg, 1.61 mmol, 88%).

LC/MS (C ₃₂ H ₄₄ N ₂ O ₅ SiS) 597 [M+H]+; RT 1.67 (ZHMS-U-B1).

1H NMR (400 MHz, DMSOO δ 7.64-7.56 (m, 4H), 7.51-7.37 (m, 6H), 4.23 (q, J=7.1 Hz, 2H), 3.60-3.47 (m, 2H), 3.44 (s , 3H), 3.19 (dd, J=14.4, 6.8 Hz, 1H), 3.00 (dd, J=14.3, 7.5 Hz, 1H), 2.06-1.94 (m, 1H), 1.54 (s, 9H), 1.24 ( t, J=7.1 Hz, 3H), 1.00 (s, 9H), 0.94 (d, J=6.7 Hz, 3H).

Step K: ethyl 2-{[(tert-butoxy)carbonyl](methyl)amino}-5-[(2S)-3-hydroxy-2-methylpropyl]-1,3thiazole-4-carboxylate.

To a solution of the product from step J (961 mg, 1.61 mmol, 1 eq.) in tetrahydrofuran (20 ml) was added tetrabutylammonium fluoride (1 M in tetrahydrofuran, 4.83 ml, 4.83 mmol, 3 eq.) and the mixture was heated with

- 54 046336 reflux for 1.5 hours. The reaction mixture was allowed to cool to ambient temperature, then partitioned between ethyl acetate and water, and the organic phase was washed with brine, dried (magnesium sulfate) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 24 g RediSep™ silica gel cartridge) eluting with a gradient of 0-80% ethyl acetate in isoheptane gave the title product as a colorless gum (204 mg, 0.57 mmol, 35%).

LC/MS (C ₁₆ H ₂₆ N ₂ O ₅ S) 359 [M+H]+; RT 1.07 (LCMS-V-BI).

1H NMR (400 MHz, DMSO<) δ 4.59 (t, J=5.2 Hz, 1H), 4.26 (q, J=7.1 Hz, 2H), 3.43 (s, 3H), 3.37-3.22 (m, 2H), 3.12 (dd, J=14.3, 6.0 Hz, 1H), 2.91 (dd, J=14.3, 8.2 Hz, 1H), 1.89-1.75 (m, 1H), 1.53 (s, 9H), 1.29 (t, J= 7.1 Hz, 3H), 0.85 (d, J=6.7 Hz, 3H).

Step L: ethyl 2-{[(tert-butoxy)carbonyl](methyl)amino}-5-[^)-3-(2-fluoro-4-iodophenoxy)-2methylpropyl]-1,3-thiazol-4- carboxylate.

To a solution of the product from step K (159 mg, 0.44 mmol, 1 eq.) in toluene (6 ml) was added 2-fluoro-4iodophenol (211 mg, 0.89 mmol, 2 eq.) followed by the addition of triphenylphosphine (233 mg, 0.89 mmol , 2 eq.) and diisopropyl azodicarboxylate (0.17 ml, 0.89 mmol, 2 eq.), and the mixture was heated at 120°C overnight. The reaction mixture was allowed to cool to ambient temperature and the solvent was removed by rotary evaporation. The resulting oil was partitioned between dichloromethane and water, and the organic phase was washed with brine, dried (magnesium sulfate) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 12 g RediSep™ silica gel cartridge) eluting with a gradient of 0-20% ethyl acetate in isoheptane gave the title product as a colorless gum (203 mg, 0.35 mmol, 79%).

LC/MS (C ₂₂ H ₂₈ FIN ₂ O5S) 579 [M+H]+; RT 1.46 (LCMS-V-BI).

1H NMR (400 MHz, DMSO-de) δ 7.59 (dd, J=10.8, 2.1 Hz, 1H), 7.48-7.42 (m, 1H), 6.97 (t, J=8.8 Hz, 1H), 4.24 (q, 2H), 3.99-3.87 (m, 2H), 3.43 (s, 3H), 3.24 (dd, J=14.2, 6.5 Hz, 1H), 3.08 (dd, J=14.3, 7.9 Hz, 1H), 2.332.20 (m, 1H), 1.51 (s, 9H), 1.27 (t, J=7.1 Hz, 3H), 1.01 (d, J=6.7 Hz, 3H).

Step M: ethyl 2-{[(tert-butoxy)carbonyl](methyl)amino}-5-[(2S)-3-{4-[3-(dimethylamino)prop-1in-1-yl]-2- fluorophenoxy}-2-methylpropyl]-1,3-thiazole-4-carboxylate.

To a solution of the product from step L (226 mg, 0.39 mmol, 1 eq.) in tetrahydrofuran (5 ml) was added 3-dimethylamino-1-propyne (0.08 ml, 0.78 mmol, 2 eq.), N.N-diisopropylethylamine (0.19 ml, 1.17 mmol, 3 eq.) and copper(1) iodide (7.44 mg, 0.04 mmol, 0.1 eq.) followed by the addition of tetrakis(triphenylphosphine)palladium(0) (45.2 mg, 0.04 mmol, 0.1 eq.), and the mixture was heated reflux for 24 hours. The reaction mixture was partitioned between ethyl acetate and water, and the organic phase was washed with brine, dried (magnesium sulfate) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 24 g RediSep™ silica gel cartridge) eluting with a gradient of 0-5% methanol in dichloromethane gave the title product as a brown gum (68 mg, 0.13 mmol, 33%).

LC/MS (C ₂ 7H ₃₆ FN3O ₅ S) 534 [M+H]+; RT 1.07 (LCMS-V-BB.

1H NMR (400 MHz, DMSO-de) δ 7.30 (dd, J=11.9, 2.0 Hz, 1H), 7.25-7.17 (m, 1H), 7.12 (t, J=8.7 Hz, 1H), 4.24 (q, 2H), 4.02-3.90 (m, 2H), 3.43 (s, 3H), 3.42 (s, 2H), 3.25 (dd, J=14.2, 6.5 Hz, 1H), 3.09 (dd, J=14.3, 8.0 Hz , 1H), 2.31-2.24 (m, 1H), 2.23 (s, 6H), 1.51 (s, 9H), 1.28 (t, J=7.1 Hz, 3H), 1.02 (d, J=6.7 Hz, 3H) .

Step N: ethyl 5-[(2S)-3-{4-[3-(dimethylamino)prop-1-in-1-yl]-2-fluorophenoxy}-2-methylpropyl]-2(methylamino)-1, 3-thiazole-4-carboxylate.

To a solution of the product from step M (68 mg, 0.13 mmol, 1 eq.) in dichloromethane (3 ml) was added trifluoroacetic acid (0.3 ml, 4.03 mmol, 31.6 eq.) and the mixture was stirred at ambient temperature overnight. The reaction mixture was diluted with dichloromethane, cooled in an ice bath, and quenched by adding saturated aqueous sodium bicarbonate. The organic phase was washed with brine, dried (magnesium sulfate) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 4 g RediSep™ silica gel cartridge) eluting with a gradient of 0-7% methanol in dichloromethane gave the title product as a brown gum (40 mg, 0.09 mmol, 72%).

LC/MS (C ₂₂ H ₂₈ FN3O3S) 434 [M+H]+; RT 0.82 (LCMS-V-BB.

1H NMR (400 MHz, DMSO-de) δ 7.52 (q, J=4.8 Hz, 1H), 7.30 (dd, J=11.9, 2.0 Hz, 1H), 7.25-7.17 (m, 1H), 7.12 (t, J=8.7 Hz, 1H), 4.19 (q, J=7.1 Hz, 2H), 4.00-3.88 (m, 2H), 3.42 (s, 2H), 3.13 (dd, J=14.5, 6.6 Hz, 1H), 3.02 (dd, J=14.4, 7.8 Hz, 1H), 2.78 (d, J=4.7 Hz, 3H), 2.23 (s, 6H), 2.21-2.12 (m, 1H), 1.25 (t, J=7.1 Hz , 3H), 1.01 (d, J=6.8 Hz, 3H).

Synthesis of 3zg: ethyl 5-(3-{4-[3-(dimethylamino)prop-1-yn-1-yl]-2-fluorophenoxy}-2-(oxan-2yloxy)propyl)-2-(methylamino)- 1,3-thiazole-4-carboxylate.

Step A: ethyl 2-{[(tert-butoxy)carbonyl](methyl)amino}-5-(prop-2-en-1-yl)-1,3-thiazol-4carboxylate.

Isopropylmagnesium chloride (2M in tetrahydrofuran; 6 mL, 12 mmol, 1.2 eq.) at -78°C was added

- 55 046336 dropwise to a solution of ethyl 5-bromo-2-[(tert-butoxycarbonyl)(methyl)amino]-1,3-thiazol-4-carboxylate (3.65 g, 9.99 mmol, 1 eq.) in dichloromethane (30 ml), and the mixture was stirred for 30 minutes. A solution of zinc chloride (1.9 M in 2-methyltetrahydrofuran; 8 mL, 15.2 mmol, 1.52 eq) was added dropwise and the mixture was stirred for 60 min at -78°C and then for 3 h at ambient temperature. The solution was cooled in ice water, and copper([)-di(lithium chloride) complex (1 M in tetrahydrofuran; 2 mL, 2 mmol, 0.2 eq.) and allyl bromide (2 mL, 23.11 mmol, 2.31 eq.) were slowly added. ). The mixture was stirred for 60 minutes at 0°C and then at ambient temperature for 18 hours. Dichloromethane (125 ml) was added and the mixture was washed successively with saturated aqueous ammonium chloride (75 ml), water (2x75 ml) and brine (75 ml), dried (magnesium sulfate) and concentrated in vacuo. Purification by flash column chromatography (50 g silica gel) eluting with a gradient of 0-10% ethyl acetate in isoheptane gave the title product as a colorless gum (1.64 g, 5.02 mmol, 50%).

LC/MS (C ₁₅ H ₂₂ N ₂ O ₄ S) 327 [M+H]+; RT 1.43 (ZHMS-U-B1).

1H NMR (400 MHz, DMSOL) δ 6.04-5.90 (m, 1H), 5.20-5.06 (m, 2H), 4.37 (q, J=7.1 Hz, 2H), 3.86 (dt, J=6.6, 1.4 Hz, 2H), 3.55 (s, 3H), 1.56 (s, 9H), 1.39 (t, J=7.1 Hz, 3H).

Step B: ethyl 5-(3-bromo-2-hydroxypropyl)-2-{[(tert-butoxy)carbonyl](methyl)amino}-1,3thiazole-4-carboxylate.

To a solution of the product from step A (1.29 g, 3.95 mmol, 1 eq.) in acetone (16 ml), cooled in ice water, was added N-bromosuccinimide (774 mg, 4.35 mmol, 1.1 eq.), ammonium acetate (30.5 mg, 0.4 mmol, 0.1 eq.) and water (4 ml), and the resulting solution was stirred for 45 min at ambient temperature. Ethyl acetate (150 ml) was added and the mixture was washed successively with water (2x75 ml) and brine (75 ml), dried (magnesium sulfate) and concentrated in vacuo. Purification by flash column chromatography (100 g silica gel) eluting with a gradient of 0-25% ethyl acetate in isoheptane gave the title product as a colorless gum (960 mg, 2.27 mmol, 57%).

LC/MS (C ₁₅ H ₂₃ BrN ₂ O ₅ S) 425 [M+H]+; RT 1.29 (ZHMS-U-B1).

1H NMR (400 MHz, DMSO-de) δ 5.59 (d, J=5.4 Hz, 1H), 4.27 (q, 2H), 3.95-3.80 (m, 1H), 3.54 - 3.45 (m, 3H), 3.44 ( s, 3H), 3.08 (dd, J=14.7, 8.3 Hz, 1H), 1.53 (s, 9H), 1.30 (t, J=7.1 Hz, 3H).

Step C: ethyl 5-[3-bromo-2-(oxan-2-yloxy)propyl]-2-{[(tert-butoxy)carbonyl](methyl)amino}1,3-thiazol-4-carboxylate.

To a solution of the product from step B (1.15 g, 2.72 mmol, 1 eq.) in dichloromethane (30 ml), cooled to 0°C, was added 3,4-dihydro-2H-pyran (1 ml, 11 mmol, 4 eq. ) followed by the addition of pyridinium ptoluenesulfonate (75 mg, 0.3 mmol, 0.11 eq.), and the mixture was stirred for 30 min at 0°C and then for 5 h at ambient temperature. Dichloromethane (30 ml) was added and the mixture was washed successively with water (2x25 ml) and brine (25 ml), dried (magnesium sulfate) and concentrated in vacuo to give the title product as a yellow gum (1.52 g, 2.7 mmol, 99% ), which was directly used in the next step without further characterization.

Step D: ethyl 5-[3-(4-bromo-2-fluorophenoxy)-2-(oxan-2-yloxy)propyl]-2-{[(tert-butoxy)carbonyl](methyl)amino}-1,3- thiazole-4-carboxylate.

4-Bromo-2-fluorophenol (0.4 mL, 3.65 mmol, 1.35 eq.) was added to a suspension of potassium tert-butoxide (400 mg, 3.56 mmol, 1.32 eq.) in dimethyl sulfoxide (5 mL) and the mixture was stirred. A solution of the product from step C (1.52 g, 2.7 mmol, 1 eq) in dimethyl sulfoxide (10 ml) was added and the mixture was stirred for 60 min at ambient temperature and then at 60°C for 3 h.

The reaction mixture was allowed to cool to ambient temperature and partitioned between ethyl acetate (150 ml) and water (75 ml), and the organic phase was washed successively with water (2x75 ml) and brine (75 ml), dried (magnesium sulfate) and concentrated in vacuo . Purification by flash column chromatography (100 g silica gel) eluting with a gradient of 0-25% ethyl acetate in isoheptane gave the title product as a pale yellow gum (1.05 g, 1.7 mmol, 63%).

LC/MS (C ₂₆ H ₃₄ BrFN ₂ O ₇ S) 617 [M+H]+; RT 1.61 (ZHMS-U-B1).

1H NMR (400 MHz, DMSO-de) δ 7.54 (dd, J=10.8, 2.4, 0.8 Hz, 1H), 7.35-7.28 (m, 1H), 7.12 (td, J=9.0, 1.1 Hz, 1H), 4.25 (q, 3H), 4.19-4.08 (m, 2H), 3.56-3.37 (m, 7H), 1.77-1.55 (m, 4H), 1.52 (d, J=2.2 Hz, 9H), 1.491.38 ( m, 4H), 1.27 (t, J=7.1 Hz, 3H).

Step E: ethyl 2-{[(tert-butoxy)carbonyl](methyl)amino}-5-(3-{4-[3-(dimethylamino)prop-1-yn-1yl]-2-fluorophenoxy}-2 -(oxan-2-yloxy)propyl)-1,3-thiazol-4-carboxylate.

To a solution of the product from step D (1.05 g, 1.7 mmol, 1 eq.) in dimethylformamide (10 ml) was added dimethyl(prop-2-yn-1-yl)amine (0.25 ml, 2.32 mmol, 1.37 eq.), iodide copper([) (32.4 mg, 0.17 mmol, 0.1 eq.) and bis(triphenylphosphine) palladium dichloride([[) (119 mg, 0.17 mmol, 0.1 eq.). N,Ndiisopropylethylamine (0.9 mL, 5.1 mmol, 3 eq.) was added and the mixture was heated at 75°C for 24 h. The reaction mixture was allowed to cool to ambient temperature and partitioned between ethyl acetate

- 56 046336 (150 ml) and water (75 ml), and the organic phase was washed successively with water (75 ml) and brine (75 ml), dried (magnesium sulfate) and concentrated in vacuo. Purification by flash column chromatography (50 g silica gel) eluting with a gradient of 0-100% ethyl acetate in isoheptane gave the title product as a brown gum (395 mg, 0.64 mmol, 38%).

LC/MS (C ₃₁ H ₄₂ FN ₃ O7S) 620 [M+H]+; RT 1.31 (ZHMS-U-B1).

Step F: ethyl 5-(3-{4-[3-(dimethylamino)prop-1-yn-1-yl]-2-fluorophenoxy}-2-(oxan-2yloxy)propyl)-2-(methylamino)- 1,3-thiazole-4-carboxylate.

A solution of the product from step E (390 mg, 0.63 mmol, 1 eq.) in 1,1,1,3,3,3-hexafluoropropan-2-ol (7 ml) was heated in a sealed tube at 100°C for 60 min , then allowed to cool to ambient temperature and concentrated in vacuo. Purification by flash column chromatography (20 g silica gel) eluting with a gradient of 0-2.5% methanol in dichloromethane gave the title product as a brown gum (170 mg, 0.33 mmol, 52%).

LC/MS (C ₂₆ H ₃₄ FN ₃ O ₅ S) 520 [M+H]+; RT 1.05 (ZHMS-U-B1).

Synthesis of 3zh: ethyl 5-(2-methoxy-3-{[tris(propan-2-yl)silyl]oxy}propyl)-2-(methylamino)-1,3thiazol-4-carboxylate.

Step A: ethyl 2-{[(tert-butoxy)carbonyl](methyl)amino}-5-(2,3-dihydroxypropyl)-1,3-thiazole-4carboxylate.

To a solution of the synthesis product 3zg, stage A (8.39 g, 25.7 mmol, 1 eq.) in a 9:1 mixture of acetone/water (250 ml) was added 4-methylmorpholine-N-oxide (4.75 g, 40.6 mmol, 1.58 eq. ) followed by the addition of osmium tetroxide (3.5 mL, 0.28 mmol, 0.01 eq.), and the resulting mixture was stirred for 18 h at ambient temperature. Water (300 ml) was added and the mixture was extracted with ethyl acetate (4x200 ml) and the combined organics were washed with brine (150 ml), dried (magnesium sulfate) and concentrated in vacuo. Purification by flash column chromatography (100 g silica gel) eluting with a gradient of 0-5% methanol in dichloromethane gave the title product as a brown gum (8.66 g, 24 mmol, 94%).

LC/MS (Ci ₅ H ₂₄ N ₂ O ₆ S) 361 [M+H]+; RT 1.07 (ZHMS-U-B1).

1H NMR (400 MHz, DMSOO δ 4.95 (d, J=5.4 Hz, 1H), 4.66 (t, J=5.6 Hz, 1H), 4.26 (q, J=7.1 Hz, 2H), 3.69-3.58 (m, 1H), 3.49-3.40 (m, 1H), 3.38-3.26 (m, 2H), 2.89 (dd, J=14.9, 8.7 Hz, 1H), 1.53 (s, 9H), 1.29 (t, J=7.1 Hz , 3H).

Step B: ethyl 2-{[(tert-butoxy)carbonyl](methyl)amino}-5-(2-hydroxy-3-{[tris(propan-2yl)silyl]oxy}propyl)-1,3-thiazole -4-carboxylate.

To a solution of the product from step A (8.65 g, 24 mmol, 1 eq.) in dichloromethane (100 ml), cooled in ice water, imidazole (3.5 g, 51.4 mmol, 2.14 eq.) was added, followed by dropwise addition of triisopropylsilyl chloride ( 6 ml, 28 mmol, 1.17 eq.), and the mixture was stirred for 60 minutes at 0°C and then for 18 hours at ambient temperature. The reaction mixture was concentrated in vacuo and purified by flash column chromatography (100 g silica gel) eluting with a gradient of 0-25% ethyl acetate in isoheptane to give the title product as a colorless oil (12.2 g, 23.5 mmol, 98%).

LC/MS (C ₂₄ H ₄₄ N ₂ O ₆ SiS) 517 [M+H]+; RT 1.69 (ZHMS-U-B1).

1H NMR (400 MHz, DMSOO δ 5.07 (d, J=5.2 Hz, 1H), 4.24 (q, J=7.1 Hz, 2H), 3.75-3.61 (m, 2H), 3.553.48 (m, 2H), 3.42 (s, 3H), 2.98 (dd, J=15.0, 8.3 Hz, 1H), 1.53 (s, 9H), 1.28 (t, J=7.1 Hz, 3H), 1.09-0.97 (m, 21H).

Step C: ethyl 2-{[(tert-butoxy)carbonyl](methyl)amino}-5-(2-methoxy-3-{[tris(propan-2yl)silyl]oxy}propyl)-1,3-thiazole -4-carboxylate.

Trimethyloxonium tetrafluoroborate (1.5 g, 10.1 mmol, 2.02 eq.) was added to a cooled solution of the product from step B (2.6 g, 5.03 mmol, 1 eq.) and KKI.I-tetramethylna(|)talin-1.8-diamine (2.2 g, 10.3 mmol, 2.04 eq.) in dichloromethane (75 ml), and the mixture was stirred at 0°C for 60 min, and then for 24 h at ambient temperature. The mixture was diluted with dichloromethane (75 ml) and washed successively with 1 M aqueous copper(II) sulfate (2x75 ml), water (75 ml) and brine (75 ml), dried (magnesium sulfate) and concentrated in vacuo. Purification by flash column chromatography (50 g silica gel) eluting with a gradient of 0-5% ethyl acetate in isoheptane gave the title product as a colorless gum (2.54 g, 4.79 mmol, 95%).

LC/MS (C ₂₅ H ₄₆ N ₂ O ₆ SiS) 531 [M+H]+; RT 1.80 (ZHMS-U-B1).

1H NMR (400 MHz, DMSOD ₆ ) δ 4.37 (q, J=7.1 Hz, 2H), 3.75 (dd, J=5.0, 2.6 Hz, 2H), 3.55 (s, 3H), 3.523.44 (m, 2H ), 3.41 (s, 3H), 3.26-3.14 (m, 1H), 1.39 (t, J=7.1 Hz, 3H), 1.09-1.01 (m, 21H).

Step D: Ethyl 5-(2-methoxy-3-{[tris(propan-2-yl)silyl]oxy}propyl)-2-(methylamino)-1,3-thiazole4-carboxylate.

A solution of the product from step C (2.54 g, 4.79 mmol, 1 eq.) in 1,1,1,3,3,3-hexafluoropropan-2-ol (20 ml) was heated in a sealed tube at 100°C for 5 h The solution was allowed to cool to ambient temperature, concentrated in vacuo and dried in vacuo to give the title product as a white solid (1.75 g, 4.06 mmol, 85%).

- 57 046336 ¹ H NMR (400 MHz, DMSO-dg) δ 5.44 (q, J=5.1 Hz, 1H), 4.35 (q, J=7.1 Hz, 2H), 3.79-3.69 (m, 2H), 3.52 ( dd, J=15.0, 4.1 Hz, 1H), 3.48-3.39 (m, 4H), 3.13 (dd, J=15.0, 7.8 Hz, 1H), 2.95 (d, J=5.0 Hz, 3H), 1.37 (t , J=7.1 Hz, 3H), 1.09 -1.01 (m, 21H).

Synthesis 4a: Na(6-chloro-4-methylpyrvdazin-3-yl)-3-(2-trimethylsilylethoxymethyl)-1,3-benzothiazol-2imine.

Step A: H-(6-chloro-4-methylpyridazin-3-yl)-1,3-benzothiazol-2-amine.

l An oven-dried single-neck, round-bottom flask was equipped with a PTFE-coated magnetic stirrer arm and a reflux condenser was installed. It was loaded with 34.0 g of 6chloro-4-methylpyridazin-3-amine (237 mmol, 1 eq.), 34 ml of 2-chloro-1,3-benzothiazole (44.2 g, 260 mmol, 1.1 eq.), 124 ml of DIPEA ( 91.8 g, 710 mmol, 3 eq.) and 137 g Cs ₂ CO ₃ (710 mmol, 3 eq.), then 1 L of DMF was added and the system was purged with argon. After 5 min of stirring in an inert atmosphere, 2.01 g of Pd ₂ (dba) ₃ (5.9 mmol, 0.025 eq.) and 6.85 g of XantPhos (11.8 mmol, 0.05 eq.) were added. The resulting mixture was then heated to 75°C and stirred at this temperature for 4 hours until complete conversion was achieved. The reaction mixture was left to cool to room temperature, then poured into 3 liters of water with vigorous stirring. After 30 min, the precipitated product was removed by filtration and then washed 2 times with water (2x2 L). The product was dried overnight under high vacuum. The dry crude product was stirred in 1 L of heptane:Et ₂ O (3:2) for 30 minutes and then filtered to obtain 64.5 g (98%) of the desired product as a green powder.

^1H NMR (500 MHz, DMSOO δ ppm 11.96 (brs, 1H), 7.86 (d, 1H), 7.65 (s, 1H), 7.51 (d, 1H), 7.38 (t, 1H), 7.21 ( t, 1H), 2.37 (s, 3H); ¹³ C NMR (125 MHz, DMSOO δ ppm 130.3, 129.5, 126.6, 122.8, 122.3, 17.2; HRMS-ERI (m/z): [M+H ]+, calculated for C ₁₂ H ₁₀ ClN ₄ S: 277.0309; found: 277.0305.

Step B: N(6-chloro-4-methylpyridazin-3-yl)-3-(2-trimethylsilylethoxymethyl)-1,3-benzothiazol-2imine.

A 2 L oven-dried single-neck, round-bottomed flask equipped with a PTFE-coated magnetic stir bar was charged with 64.5 g of the product from step A (236 mmol, 1 eq.), 123 ml of DIPEA (9.16 g, 708 mmol, 3 eq. ) and 14.43 g of N-dimethylpyridin-4-amine (11.81 mmol, 0.05 eq.) in 1 L of dry DCM, and cooled to 0°C in an N2 atmosphere. With vigorous mechanical stirring, 46.00 mL of 2-(chloromethoxy)ethyltrimethylsilane (43.32 g, 259 mmol, 1.1 eq.) was added dropwise to the mixture over a period of 5 min. The mixture was stirred at 0°C for 30 minutes, during which time the reaction reached a state of complete conversion. 24.5 ml of water was added to the reaction mixture, then celite was added, and the volatiles were removed under reduced pressure. The crude product was purified by flash column chromatography using heptane and EtOAc as eluents to obtain 46.62 g (48%) of the desired product.

¹ H NMR (500 MHz, DMSO^6) δ ppm 7.85 (dm, 1H), 7.72 (q, 1H), 7.53 (dm, 1H), 7.47 (m, 1H), 7.29 (m, 1H), 5.89 (s, 2H), 3.70 (m, 2H), 2.39 (d, 3H), 0.90 (m, 2H), -0.12 (s, 9H); ¹³ C NMR (125 MHz, DMSO^) δ ppm 159.5, 158.5, 150.0, 138.1, 137.4, 129.5, 127.4, 125.5, 123.8, 123.2, 112.4, 73.0, 66.8, 17.7, 17.1, -1.0; MSVRERI (m/z): [M+H]+, calculated for C ₁₈ H ₂₄ ClN ₄ OSSi: 407.1123; found: 407.1120.

Synthesis 4b: (22)-L-(6-Chloropyridazin-3-yl)-3-{[2-(trimethylsilyl)ethoxy]methyl}-2,3-dihydro-1,3benzothiazol-2-imine.

N.N-Diisopropylethylamine (3 ml, 16.6 mmol, 3 equiv.) was added under nitrogen atmosphere to a suspension of synthesis product 9b (1.45 g, 5.52 mmol, 1 equiv.) in dichloromethane (75 ml) at 0°C. [2(chloromethoxy)ethyl]trimethylsilane (1.2 mL, 6.78 mmol, 1.23 eq.) and 4-dimethylaminopyridine (33.7 mg, 0.28 mmol, 0.05 eq.) were added and the mixture was stirred at room temperature for 3 h. Dichloromethane ( 75 ml), and the mixture was washed with water (2x75 ml) and brine (75 ml). The solution was dried (magnesium sulfate) and concentrated in vacuo. Trituration with methanol (20 ml) resulted in a solid, which was filtered, washed with methanol (2x10 ml) and dried in vacuo to give the title product as a pale brown solid (1.85 g, 4.71 mmol, 85%).

LC/MS (C ₁₇ H ₂₁ ClN ₄ OSiS) 393 [M+H]+; RT 1.56 (ZHMS-U-B1).

Synthesis 5a: methyl 5-[3-[4-[3-[tert-butoxycarbonyl(methyl)amino]prop-1-ynyl]-2fluorophenoxy]propyl]-2-[[4-[tert-butyl(diphenyl)silyl ]oxy-5-(p-tolylsulfonyloxy)pentyl]-[5methyl-6-[(2)-[3 -(2-trimethylsilylethoxymethyl)-1,3-benzothiazol-2-ylidene]amino]pyridazin-3 yl]amino ]thiazole-4-carboxylate.

Step A: methyl 5-[3-[4-[3-[tert-butoxycarbonyl(methyl)amino]prop-1-ynyl]-2fluorophenoxy]propyl]-2-[[5-[tert-butyl(dimethyl)silyl ]oxy-4-[tert-butyl(diphenylsilyl]oxypentyl][5-methyl-6-[(2)-[3-(2-trimethylsilylethoxymethyl)-1,3-benzothiazol-2-ylidene]amino]pyridazin-3yl ]amino]thiazole-4-carboxylate.

Using the general Buchwald reaction procedure II, starting from 12 g of synthesis product 3a (13 mmol) and 6.30 g of synthesis product 4a (15.6 mmol) as a suitable halide, 14 g (83%) of the desired product were obtained.

^1H NMR (500 MHz, DMSO^ ₆ ) δ ppm 7.85-7.23 (m, 14H), 7.58 (s, 1H), 7.31 (t, 1H), 7.19 (m, 1H), 7.14 (t, 1H), 5.86 (s, 2H), 4.37 (t, 2H) , 4.20 (s, 2H), 4.15 (t, 2H), 3.73 (s, 3H), 3.71 (t, 2H), 3.67 (m, 1H), 3.39 (m, 2H),

- 58 046336

3.27 (t, 2H), 2.83 (s, 3H), 2.41 (s, 3H), 2.12 (m, 2H), 1.72 (m, 2H), 1.52 (m, 2H), 1.40 (s, 9H), 0.90 (t, 2H), 0.89 (s, 9H), 0.69 (s, 9H), -0.14 (s, 9H), -0.19/-0.23 (s+s, 6H); ¹³ C NMR (125 MHz, DMSOL ₆ ) δ ppm 147.5, 129.1, 119.3, 117.5, 115.4, 73.4, 72.3, 68.4, 66.8, 65.8, 51.8, 46.6, 38.5, 33.8, 31.0, 30.5, 28.5, 27.1, 26.1, 23.0 , 22.6, 17.9, 17.8, -1.0, - 5.3; MSHR-ERI (m/z): [M+H] ⁺ , calculated for C68H93FN?O8S2Si3: 1302.5813; found: 1302.5819.

Step B: methyl 5-[3-[4-[3-[tert-butoxycarbonyl(methyl)amino]prop-1-ynyl]-2fluorophenoxy]propyl]-2-[[4-[tert-butyl(diphenyl)silyl ]oxy-5-hydroxypentyl]-[5-methyl-6-[(2)-[3-(2trimethylsilylethoxymethyl)-1,3-benzothiazol-2-ylidene]amino]pyridazin-3-yl]amino]thiazol-4carboxylate .

A 100 mL oven-dried, single-neck, round-bottom flask was equipped with a PTFE-coated magnetic stir bar and a reflux condenser was installed. It was loaded with 1.40 g of the product from step A (1.1 mmol, 1 eq.) and 12 mg of camphorsulfonic acid (0.054 mmol, 0.05 eq.), 5 ml of DCM and 1 ml of MeOH. The resulting mixture was stirred overnight at room temperature. until complete transformation is achieved. The reaction mixture was directly concentrated on Celite and then purified by flash column chromatography using heptane and EtOAc as eluents to obtain 700 mg (55%) of the title product as a yellow solid.

1H NMR (500 MHz, DMSOHO δ ppm 7.85-7.14 (m, 14H), 7.56 (s, 1H), 7.32 (dd 1H), 7.20 (m, 1H), 7.15 (t, 1H), 5.86 ( s, 2H), 4.56 (t, 1H), 4.33 (m, 2H), 4.20 (s, 2H), 4.15 (t, 2H), 3.74 (s, 3H), 3.72 (t, 2H), 3.65 (m , 1H), 3.27 (t, 2H), 3.27 (t, 2H), 2.83 (s, 3H), 2.41 (s, 3H), 2.13 (m, 2H), 1.73/1.64 (m+m, 2H), 1.52 (m, 2H), 1.40 (s, 9H), 0.90 (t, 2H), 0.86 (s, 9H), -0.13 (s, 9H); ¹³ C NMR (125 MHz, DMSO^) δ ppm . 154.9, 147.6, 129.1, 119.4, 117.5, 115.4, 82.4, 73.7, 72.9, 68.4, 66.8, 64.5, 51.9, 46.8, 38.5, 33.8, 31.0, 30.6, 28.5, 2 7.2, 23.1, 22.5, 17.9, 17.8, - 1.0; HRMS-ERI (m/z): [M+H] ⁺ , calculated for C ₆₂ H ₇₉ FN ₇ O ₈ S ₂ Si ₂ : 1188.4949; found: 1188.4938.

Step C: methyl 5-[3-[4-[3-[tert-butoxycarbonyl(methyl)amino]prop-1-ynyl]-2fluorophenoxy]propyl]-2-[[4-[tert-butyl(diphenyl)silyl ]oxy-5-(p-tolylsulfonyloxy)pentyl]-[5methyl-6-[(2)-[3 -(2-trimethylsilylethoxymethyl)-1,3-benzothiazol-2-ylidene]amino]pyridazin-3 yl]amino ]thiazole-4-carboxylate.

A 100 mL oven-dried, single-neck, round-bottomed flask was equipped with a PTFE-coated magnetic stirrer arm and loaded with 700 mg of the product from step B (0.58 mmol, 1 eq.) and 907 mg of N-dimethyl-1-(p-tolylsulfonyl chloride )pyridin-1-ium-4-amine (2.9 mmol, 5 eq., Tetrahedron Lett. 2016, 57, 4620), dissolved them in 35 ml of DCM and the solution was stirred overnight at room temperature. The reaction has reached a state of complete transformation. The reaction mixture was concentrated directly on celite, and then purified by flash column chromatography using heptane and EtOAc as eluents to obtain 450 mg (56%) of the desired product.

1H NMR (500 MHz, DMSOO δ ppm 7.88-7.23 (m, 14H), 7.58 (m, 2H), 7.53 (s, 1H), 7.31 (m, 2H), 7.31 (dd, 1H), 7.19 (m, 1H), 7.15 (t, 1H), 5.86 (s, 2H), 4.20 (s, 2H), 4.16 (t, 2H), 4.15 (t, 2H), 3.92 (m, 2H), 3.84 ( m, 1H), 3.72 (t, 2H), 3.70 (s, 3H), 3.27 (t, 2H), 2.83 (s, 3H), 2.41 (s, 3H), 2.33 (s, 3H), 2.13 (m , 2H), 1.47 (m, 2H), 1.47 (m, 2H), 1.40 (s, 9H), 0.91 (t, 2H), 0.86 (s, 9H), -0.13 (s, 9H); ¹³ C NMR (125 MHz, DMSOL6) δ ppm 147.5, 145.3, 130.4, 129.1, 128.0, 119.3, 117.4, 115.5, 72.9, 72.6, 70.4, 68.4, 66.8, 51.8, 46.2, 38.6 , 33.8, 31.0, 30.1, 28.5 , 27.0, 23.1, 22.4, 21.5, 17.8, 17.8, -1.0; MSHR-ERI (m/z): [M+H] ⁺ , calculated for C69H85FN7O10S3Si2: 1342.5037; found: 1342.5039.

Synthesis 5b: methyl 2-[[4-tert-butyl(diphenyl)silyl]oxy-5-(p-tolylsulfonyloxy)pentyl]-[5methyl-6-[(2)-[3-(2-trimethylsilylethoxymethyl)-1 ,3-benzothiazol-2-ylidene]amino] pyridazin-3 -yl]amino] 5-[3-[4-[3-(dimethylamino)prop-1-ynyl]-2-fluorophenoxy]propyl]thiazol-4- carboxylate.

Step A: methyl 2-[[5-[tert-butyl(dimethyl)silyl]oxy-4-[tert-butyl(diphenyl)silyl]oxypentyl][5-methyl-6-[(2)-[3-( 2-trimethylsilylethoxymethyl)-1,3-benzothiazol-2-ylidene]amino]pyridazin-3yl]amino]-5-[3-[4-[3-(dimethylamino)prop-1-ynyl]-2-fluorophenoxy]propyl ]thiazole-4-carboxylate.

Using the general Buchwald reaction procedure II, starting from 6.0 g of synthesis product 3b (7.1 mmol, 1 eq.) and 3.46 g of synthesis product 4a (8.51 mmol, 1.2 eq.) as the appropriate halide, 7.0 g (81%) of the target product was obtained product.

1H NMR (500 MHz, DMSOL ₆ ) δ ppm 7.84-7.25 (m, 4H), 7.60-7.11 (m, 10H), 7.58 (s, 1H), 7.29 (t, 1H), 7.19 (dd, 1H), 7.14 (m, 1H), 5.86 (s, 2H), 4.37 (t, 2H), 4.15 (t, 2H), 3.74 (s, 3H), 3.71 (t, 2H), 3.67 (m, 1H), 3.39 (d, 2H), 3.38 (s, 2H ), 3.27 (t, 2H), 2.41 (s, 3H), 2.19 (s, 6H), 2.13 (m, 2H), 1.73 (m, 2H), 1.51 (m, 2H), 0.91 (t, 2H) , 0.89 (s, 9H), 0.69 (s, 9H), -0.13 (s, 9H), -0.19/-0.23 (s+s, 6H); ¹³ C NMR (125 MHz, DMSOL6) δ ppm 135.9-111.3, 127.2-112.0, 128.9, 119.2, 117.6, 115.5, 73.4, 72.9, 68.4, 66.8, 65.9, 51.9, 48.1, 46.6, 44.2, 31.0, 30.9, 27. 3, 26.0, 23.1, 22.6, 17.9, 17.8, -1.0, -5.3; MSHR-ERI (m/z): [M+H] ⁺ , calculated for C64H87FN ₇ O6S2Si3: 1216.5446; found: 1216.5425.

Step B: methyl 2-[[4-[tert-butyl(diphenyl)silyl]oxy-5-hydroxypentyl]-[5-methyl-6-[(2)-[3-(2trimethylsilylethoxymethyl)-1,3-benzothiazole -2-ylidene]amino]pyridazin-3-yl]amino]-5-[3-[4-[3(dimethylamino)prop-1-ynyl]-2-fluorophenoxy]propyl]thiazole-4-carboxylate.

A 100 ml oven-dried single-necked, round-bottomed flask was equipped with a mag anchor

- 59 046336 thread stirrer coated with PTFE, and a reflux condenser was installed. It was loaded with 7.0 g of the product from stage A (5.75 mmol, 1 eq.) and 1.60 g of camphorsulfonic acid (6.90 mmol, 1.2 eq.), dissolved in 25 ml of DCM and 5 ml of MeOH. The resulting mixture was stirred overnight at room temperature. until complete transformation is achieved. The reaction mixture was concentrated directly on celite, and purified by flash column chromatography using DCM and MeOH (1.2% NH ₃ ) as eluents to obtain 3.0 g (47%) of the title product as a yellow solid.

Step C: methyl 2-[[4-[tert-butyl(diphenyl)silyl]oxy-5-(p-tolylsulfonyloxy)pentyl]-[5methyl-6-[(2)-[3 -(2-trimethylsilylethoxymethyl)- 1,3-benzothiazol-2-ylidene]amino] pyridazin-3 -yl]amino] 5-[3-[4-[3-(dimethylamino)prop-1-ynyl]-2-fluorophenoxy]propyl]thiazol-4 -carboxylate.

A 100 mL oven-dried single-neck, round-bottomed flask was equipped with a PTFE-coated magnetic stirrer armature and charged with 3.80 g of the product from step B (3.40 mmol, 1 eq.), 5.40 g of M,M-dimethyl-1-(p-tolylsulfonyl) chloride )pyridinium-1-ium-4-amine (17.0 mmol, 5 eq., Tetrahedron Lett. 2016, 57, 4620) and 20 ml DCM, and then stirred overnight at RT. The reaction has reached a state of complete transformation. The reaction mixture was directly concentrated on celite, and then purified by flash column chromatography using DCM and MeOH (1.2% NH ₃ ) as eluents to obtain 3.58 g (83%) of the title product.

¹ H NMR (500 MHz, DMSOO δ ppm (m, 10H), 7.85 (d, 1H), 7.59 (d, 2H), 7.53 (m, 1H), 7.47 (d, 1H), 7.44 (t , 1H), 7.32 (d, 2H), 7.3 (dd, 1H), 7.26 (t, 1H), 7.20 (d, 1H), 7.15 (t, 1H), 5.87 (s, 2H), 4.16 (t, 2H), 4.16 (br., 2H), 3.92 (m, 2H), 3.84 (m, 1H), 3.72 (t, 2H), 3.70 (s, 3H), 3.38 (s, 2H), 3.27 (t, 2H), 2.42 (s, 3H), 2.33 (s, 3H), 2.19 (s, 6H), 2.13 (m, 2H), 1.47 (br., 2H), 1.47 (br., 2H), 0.91 (t , 2H), 0.87 (s, 9H), -0.13 (s, 9H); ¹³ C NMR (125 MHz, DMSO-de) δ ppm 130.5, 128.9, 128.0, 127.2, 123.5, 123.2, 119.2, 117.4 , 115.5, 112.0, 72.9, 72.6, 70.4, 68.4, 66.8, 51.8, 48.1, 46.1, 44.2, 31.0, 30.1, 27.0, 23.1, 22.4, 21.5, 17.8, 17.8, -1.0; MSVR-ERI (m/z) : [M+H]+, calculated for C ₆₅ H ₇ 9FN ₇ O ₈ S3Si ₂ : 1256.4669; found: 1256.4677.

Synthesis of 5c: methyl 5-[3-[4-[3-[tert-butoxycarbonyl(methyl)amino]prop-1-ynyl]-2fluorophenoxy]propyl]-2-[4-hydroxybutyl-[5-methyl-6- [(2)-[3-(2-trimethylsilylethoxymethyl)-1,3benzothiazol-2-ylidene]amino]pyridazin-3-yl]amino]thiazol-4-carboxylate.

Step A: methyl 5-[3-[4-[3-[tert-butoxycarbonyl(methyl)amino]prop-1-ynyl]-2fluorophenoxy]propyl]-2-[4-[tert-butyl(dimethyl)silyl] hydroxybutyl-[5-methyl-6-[(2)-[3-(2trimethylsilylethoxymethyl)-1,3-benzothiazol-2-ylidene]amino]pyridazin-3-yl]amino]thiazol-4carboxylate.

Using the general Buchwald reaction procedure II, starting from 250 mg of synthesis product 3c (0.38 mmol, 1 eq.) and 250 mg of synthesis product 4a (0.38 mmol, 1 eq.) as a suitable halide, 295 mg (75%) of the target product were obtained product.

1H NMR (500 MHz, DMSOO δ ppm 7.83-7.25 (m, 4H), 7.64 (s, 1H), 7.31 (dd, 1H), 7.21 (m, 1H), 7.15 (t, 1H), 5.85 (s, 2H), 4.41 (t, 2H), 4.20 (s, 2H), 4.14 (t, 2H), 3.77 (s, 3H), 3.71 (t, 2H), 3.61 (t, 2H), 3.26 ( t, 2H), 2.84 (s, 3H), 2.44 (s, 3H), 2.12 (m, 2H), 1.74 (m, 2H), 1.51 (m, 2H), 1.40 (s, 9H), 0.91 (t , 2H), 0.79 (s, 9H), 0.03 (s, 6H), -0.12 (s, 9H); ¹³ C NMR (125 MHz, DMSOD6) δ ppm 147.5, 137.6, 129.1, 127.2-112, 119.3, 117.7, 115.5, 72.8, 68.4, 66.8, 62.5, 51.9, 46.7, 38.3, 33.8, 31.0, 29.8, 28.5, 26.2, 23.8, 23.1, 17.9, 17.8, -1.0, -4 .9; MSVR-ERI (m/ z): [M+H]+, calculated for C51H ₇ 3FN ₇ O ₇ S2Si2: 1034.4530; found: 1034.4517.

Step B: methyl 5-[3-[4-[3-[tert-butoxycarbonyl(methyl)amino]prop-1-ynyl]-2fluorophenoxy]propyl]-2-[4-hydroxybutyl-[5-methyl-6- [(2)-[3-(2-trimethylsilylethoxymethyl)-1,3benzothiazol-2-ylidene]amino]pyridazin-3-yl]amino]thiazol-4-carboxylate.

Using the general procedure for deprotection of tert-butyldimethylsilyl, starting from 650 mg of the product from step A (0.63 mmol, 1 eq.) and 36 mg of [(1S,4R)-7,7-dimethyl-2-oxonorbornan-1yl]methanesulfonic acid (0.16 mmol, 0.25 eq.), 178 mg (31%) of the target product were obtained.

1H NMR (500 MHz, DMSOO δ ppm 7.84 (d, 1H), 7.68 (s, 1H), 7.46 (d, 1H), 7.43 (td, 1H), 7.32 (brd., 1H), 7.25 ( td, 1H), 7.22 (d, 1H), 7.17 (t, 1H), 5.86 (s, 2H), 4.39 (t, 2H), 4.20 (br., 2H), 4.15 (t, 2H), 3.76 ( s, 3H), 3.72 (t, 2H), 3.45 (q, 2H), 3.26 (t, 2H), 2.84 (br., 3H), 2.45 (s, 3H), 2.12 (m, 2H), 1.72 ( m, 2H), 1.50 (m, 2H), 1.40 (s, 9H), 0.92 (t, 2H), -0.11 (s, 9H); ¹³ C NMR (125 MHz, DMSO-de) δ ppm. 129.1, 127.2, 123.4, 123.2, 119.3, 117.6, 115.5, 111.9, 72.9, 68.4, 66.7, 60.9, 52.0, 46.8, 38.5, 33.8, 31.0, 29.9, 28.5, 2 4.1, 23.2, 17.9, 17.8, 1.0; MSVR- ERI (m/z): [M+H]+, calculated for C ₄ 5H ₅₉ FN ₇ O ₇ S ₂ Si: 920.3665; found: 920.3650.

Synthesis 5d: methyl 5-[3-[4-[3-(dimethylamino)prop-1-ynyl]-2-fluorophenoxy]propyl]-2-[4hydroxybutyl-[5-methyl-6-[(2)-[ 3-(2-trimethylsilylethoxymethyl)-1,3-benzothiazol-2ylidene]amino]pyridazin-3-yl]amino]thiazol-4-carboxylate.

Step A: methyl 2-[4-[tert-butyl(dimethyl)silyl]oxybutyl-[5-methyl-6-[(2)-[3-(2trimethylsilylethoxymethyl)-1,3-benzothiazol-2-ylidene]amino ]pyridazin-3-yl]amino]-5-[3-[4-[3(dimethylamino)prop-1-ynyl]-2-fluorophenoxy]propyl]thiazole-4-carboxylate.

Using the general Buchwald reaction procedure II, starting from 1800 mg of synthesis product 3d (3.11 mmol, 1 eq.) and 1267 mg of synthesis product 4a (3.11 mmol, 1 eq.) as the appropriate halide, 2035 mg (69%) of the target product were obtained product.

1H NMR (500 MHz, DMSOO δ ppm 7.82 (dm, 1H), 7.61 (s, 1H), 7.44 (dm, 1H), 7.42 (m, 1H), 7.29 (dd,

- 60 046336

1H), 7.23 (m, 1H), 7.19 (dm, 1H), 7.13 (t, 1H), 5.83 (s, 2H), 4.40 (t, 2H), 4.13 (t, 2H), 3.77 (s, 3H ), 3.70 (m, 2H), 3.60 (t, 2H), 3.38 (s, 2H), 3.25 (m, 2H), 2.42 (s, 3H), 2.19 (s, 6H), 2.11 (m, 2H) , 1.73 (m, 2H), 1.51 (m, 2H), 0.90 (m, 2H), 0.78 (s, 9H), -0.03 (s, 6H), -0.13 (s, 9H); ¹³ C NMR (125 MHz, DMSO-dg) δ ppm 128.9, 127.1, 123.4, 123.2, 119.2, 117.5, 115.4, 111.9, 72.8, 68.4, 66.7, 62.5, 51.9, 48.1, 46.5, 44.2, 30.9, 29.7, 26.2, 2 3.7, 23.1, 17.9, 17.8, -1.0, - 4.9; MSHR-ERI (m/z): [M+H]+, calculated for C ₄₇ H ₆₇ FN ₇ O ₅ S ₂ Si ₂ : 948.4162; found: 948.4161.

Step B: methyl 5-[3-[4-[3-(dimethylamino)prop-1-ynyl]-2-fluorophenoxy]propyl]-2-[4hydroxybutyl-[5-methyl-6-[(2)-[ 3-(2-trimethylsilylethoxymethyl)-1,3-benzothiazol-2ylidene]amino]pyridazin-3-yl]amino]thiazol-4-carboxylate.

Using the general procedure for deprotection of the tert-butyldimethylsilyl protecting group, starting from 7.23 g of the product from step A (7.63 mmol, 1 eq.) and 2.22 g of [(1S,4R)-7,7-dimethyl-2-oxonorbornan-1yl]methanesulfonic acid (9.54 mmol, 1.25 eq.) gave 5.23 g (82%) of the target product.

1H NMR (500 MHz, DMSO-de) δ ppm 7.84 (d, 1H), 7.69 (s, 1H), 7.47 (d, 1H), 7.44 (td, 1H), 7.35 (dd, 1H), 7.25 (td, 1H), 7.25 (dm, 1H), 7.18 (t, 1H), 5.86 (s, 2H), 4.46 (t, 1H), 4.39 (t, 2H), 4.15 (t, 2H), 3.78 (s, 3H), 3.72 (m, 2H), 3.65 ( s, 2H), 3.45 (q, 2H), 3.27 (t, 2H), 2.46 (s, 3H), 2.39 (s, 6H), 2.13 (m, 2H), 1.72 (m, 2H), 1.50 (m , 2H), 0.92 (m, 2H), -0.11 (s, 9H); ¹³ C NMR (125 MHz, DMSOO δ ppm 163.2, 129.2, 127.2, 123.4, 123.2, 119.3, 117.6, 115.5, 112.0, 72.9, 68.4, 66.8, 60.9, 52.0, 47. 8, 46.8, 43.6, 31.0, 29.9 , 24.1, 23.1, 17.9, 17.8, -0.9; MSHR-ERI (m/z): [M+H]+, calculated for C ₄₁ H ₅ 3FN ₇ O ₅ S ₂ Si: 834.3297; found: 834.3296.

Synthesis 5e: methyl 5-[3-[4-[3-[tert-butoxycarbonyl(methyl)amino]prop-1-ynyl]-2fluorophenoxy]propyl]-2-[5-hydroxypentyl-[5-methyl-6- [(2)-[3-(2-trimethylsilylethoxymethyl)-1,3benzothiazol-2-ylidene]amino]pyridazin-3-yl]amino]thiazol-4-carboxylate.

Step A: methyl 5-[3-[4-[3-[tert-butoxycarbonyl(methyl)amino]prop-1-ynyl]-2fluorophenoxy]propyl]-2-[5-[tert-butyl(dimethyl)silyl] oxypentyl-[5-methyl-6-[(2)-[3-(2trimethylsilylethoxymethyl)-1,3-benzothiazol-2-ylidene]amino]pyridazin-3-yl]amino]thiazol-4carboxylate.

Using the general Buchwald reaction procedure II, starting from 350 mg of synthesis product 3m (0.52 mmol, 1 eq.) and 210 mg of synthesis product 4a (0.52 mmol, 1 eq.) as the appropriate halide, 481 mg (88%) of the target product were obtained product.

1H NMR (500 MHz, DMSOA) δ ppm 7.81 (dd, 1H), 7.62 (s, 1H), 7.45 (dd, 1H), 7.42 (m, 1H), 7.28 (dd, 1H), 7.24 (m, 1H), 7.19 (m, 1H), 7.15 (t, 1H), 5.84 (s, 2H), 4.38 (t, 2H), 4.20 (s, 2H), 4.16 (t, 2H), 3.78 (s, 3H), 3.73 (t, 2H), 3.55 ( t, 2H), 3.26 (t, 2H), 2.85 (s, 3H), 2.44 (s, 3H), 2.13 (m, 2H), 1.71 (m, 2H), 1.51 (m, 2H), 1.41 (s , 9H), 1.40 (m, 2H), 0.92 (t, 2H), 0.80 (s, 9H), -0.04 (s, 6H), -0.10 (s, 9H); ¹³ C NMR (125 MHz, DMSOO δ ppm 163.2, 147.5, 137.6, 129.0, 127.1, 123.5, 123.1, 119.3, 117.7, 115.7, 111.9, 73.0, 68.6, 66.8, 6 2.7, 51.8, 47.0, 38.6, 33.8 , 32.4, 31.0, 28.5, 27.1, 26.2, 23.1, 23.0, 17.9, 17.8, -1.0, -5.0; MSVR-ERI (m/z): [M+H]+, calculated for C ₅₂ H ₇₅ FN ₇ O ₇ S ₂ Si ₂ : 1048.4686; found: 1048.4692.

Step B: methyl 5-[3-[4-[3-[tert-butoxycarbonyl(methyl)amino]prop-1-ynyl]-2fluorophenoxy]propyl]-2-[5-hydroxypentyl-[5-methyl-6- [(2)-[3-(2-trimethylsilylethoxymethyl)-1,3benzothiazol-2-ylidene]amino]pyridazin-3-yl]amino]thiazol-4-carboxylate.

Using the general procedure for removing the tert-butyldimethylsilyl protecting group, starting from 2.52 g of the product from step A (2.40 mmol, 1 eq.) and 0.70 g of |(u,4K)-7,7-dimethyl-2-oxonorbornan-1yl]methanesulfonic acid (3.00 mmol, 1.25 eq.) gave 1.19 g (53%) of the target product.

1H NMR (500 MHz, DMSOD ₆ ) δ ppm 7.84 (d, 1H), 7.66 (s, 1H), 7.46 (d, 1H), 7.43 (t, 1H), 7.32 (d, 1H), 7.25 (t, 1H), 7.22 (d, 1H), 7.16 (t, 1H), 5.86 (s, 2H), 4.36 (t, 2H), 4.20 (s, 2H), 4.15 (t, 2H), 3.78 (s, 3H), 3.72 (t, 2H), 3.4 ( t, 2H), 3.26 (t, 2H), 2.84 (s, 3H), 2.46 (s, 3H), 2.12 (qn, 2H), 1.69 (qn, 2H), 1.49 (m, 2H), 1.40 (s , 9H), 1.40 (qn, 2H), 0.92 (t, 2H), -0.11 (s, 9H); ¹³ C NMR (125 MHz, DMSOO δ ppm 129.1, 127.2, 123.4, 123.2, 119.3, 117.6, 115.5, 112.0, 72.9, 68.4, 66.7, 61.0, 52.0, 47.0, 38.5 , 33.9, 32.5, 31.0, 28.5 , 27.1, 23.2, 23.1, 17.9, 17.8, -1.0; HRMS-ERI (m/z): [M+Na]+, calculated for C46H6cFN ₇ NaO ₇ S2Si: 956.3641; found: 956.3646.

Synthesis of 5f: methyl 5-[3-[4-[3-(dimethylamino)prop-1-ynyl]-2-fluorophenoxy]propyl]-2-[5hydroxypentyl-[5-methyl-6-[(2)-[ 3-(2-trimethylsilylethoxymethyl)-1,3-benzothiazol-2ylidene]amino]pyridazin-3-yl]amino]thiazol-4-carboxylate.

Step A: methyl 2-[5-[tert-butyl(dimethyl)silyl]oxypentyl-[5-methyl-6-[(2)-[3-(2trimethylsilylethoxymethyl)-1,3-benzothiazol-2-ylidene]amino ]pyridazin-3-yl]amino]-5-[3-[4-[3(dimethylamino)prop-1-ynyl]-2-fluorophenoxy]propyl]thiazole-4-carboxylate.

Using the general Buchwald reaction procedure II, starting from 3.68 g of synthesis product 3t (6.22 mmol, 1 eq.) and 3.29 g of synthesis product 4a (8.08 mmol, 1.3 eq.) as the appropriate halide, 3.92 g (65%) of the target product was obtained product.

1H NMR (500 MHz, DMSOD ₆ ) δ ppm 7.83 (dm, 1H), 7.66 (s, 1H), 7.46 (dm, 1H), 7.43 (m, 1H), 7.30 (dd, 1H), 7.25 (m, 1H), 7.15 (dm, 1H), 5.85 (s, 2H), 4.40 (t, 2H), 4.14 (t, 2H), 4.14 (t, 1H), 3.77 (s, 3H), 3.71 (m, 2H), 3.54 (t, 2H), 3.38 ( s, 2H), 3.26 (t, 2H), 2.44 (s, 3H), 2.19 (s, 6H), 2.12 (m, 2H), 1.70 (m, 2H), 1.49 (m, 2H), 1.37 (m , 2H), 0.91 (m, 2H), 0.79 (s, 9H), -0.06 (s, 6H), -0.12 (s, 9H); MSHR-ERI (m/z): [M+H]+, calculated for C ₄₈ H ₆₉ FN ₇ O ₅ S ₂ Si ₂ : 962.4319; found: 962.4301.

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Step B: methyl 5-[3-[4-[3-(dimethylamino)prop-1-ynyl]-2-fluorophenoxy]propyl]-2-[5hydroxypentyl-[5-methyl-6-[(2)-[ 3-(2-trimethylsilylethoxymethyl)-1,3-benzothiazol-2ylidene]amino]pyridazin-3-yl]amino]thiazol-4-carboxylate.

Using the general procedure for deprotection of the tert-butyldimethylsilyl protecting group, starting from 3.91 g of the product from step A (4.06 mmol, 1 eq.) and 1.18 g of [(1S,4R)-7,7-dimethyl-2-oxonorbornan-1yl]methanesulfonic acid (5.08 mmol, 1.25 eq.) gave 2.61 g (76%) of the target product.

Ή NMR (500 MHz, DMSO-de) δ ppm 7.84 (dm, 1H), 7.67 (s, 1H), 7.47 (dm, 1H), 7.42 (td, 1H), 7.31 (dd, 1H), 7.25 (td, 1H), 7.21 (dd, 1H), 7.16 (t, 1H), 5.86 (s, 2H), 4.37 (m, 1H), 4.37 (m, 2H), 4.14 (t, 2H), 3.78 (s, 3H), 3.72 (t, 2H), 3.40 ( m, 2H), 3.38 (s, 2H), 3.27 (m, 2H), 2.46 (s, 3H), 2.20 (s, 6H), 2.12 (m, 2H), 1.69 (m, 2H), 1.49 (m , 2H), 1.39 (m, 2H), 0.92 (t, 2H), -0.11 (s, 9H); ¹³ C NMR (125 MHz, DMSOO δ ppm 129.0, 127.2, 123.5, 123.2, 119.2, 117.6, 115.5, 111.9, 72.9, 68.4, 66.7, 61.0, 52.0, 48.1, 47.0 , 44.2, 32.5, 31.0, 27.1 , 23.1, 23.1, 17.8, 17.8, -0.9; MSHR-ERI (m/z): [M+2H]2+, calculated for C42H56FN-O5S2Si: 424.6764; found: 424.6755.

Synthesis of 5g: ethyl 5-(3-iodopropyl)-2-[methyl-[5-methyl-6-[(2)-[3-(2-trimethylsilylethoxymethyl)-1,3benzothiazol-2-ylidene]amino]pyridazin- 3-yl]amino]thiazol-4-carboxylate.

Step A: ethyl 5-(3-chloropropyl)-2-[methyl-[5-methyl-6-[(2)-[3-(2-trimethylsilylethoxymethyl)-1,3benzothiazol-2-ylidene]amino]pyridazin- 3-yl]amino]thiazol-4-carboxylate.

Using the general procedure II of the Buchwald reaction, starting from 3.15 g of the synthesis product 3 (12 mmol, 1.2 eq.) and 4.07 g of the synthesis product 4a (10 mmol, 1 eq.) as a suitable halide, we obtained 2.6 g (41%) of the target product.

¹ H NMR (500 MHz, DMSOO δ ppm 7.84 (d, 1H), 7.65 (s, 1H), 7.45 (d, 1H), 7.43 (tm, 1H), 7.25 (tm, 1H), 5.85 ( s, 2H), 4.30 (q, 2H), 3.77 (s, 3H), 3.71 (t, 2H), 3.71 (t, 2H), 3.22 (t, 2H), 2.48 (s, 3H), 2.10 (quin , 2H), 1.31 (t, 3H), 0.92 (t, 2H), -0.11 (s, 9H); ¹³ C NMR (125 MHz, DMSO^) δ ppm 162.6, 157.4, 156.8, 155.1, 151.7 , 140.5, 137.6, 137.1, 135.3, 125.6, 123.5, 123.2, 123.1, 117.6, 111.9, 72.9, 66.7, 60.7, 45.3, 35.4, 34.4, 24.3, 18.0, 17. 8, 14.7, -1.0; MSVR-ERI (m/ z): [M+H]+, calculated for C ₂₈ H ₃₈ ClN ₆ O ₃ S ₂ Si: 633.1899; found: 633.1891.

Step B: ethyl 5-(3-iodopropyl)-2-[methyl-[5-methyl-6-[(2)-[3-(2-trimethylsilylethoxymethyl)-1,3benzothiazol-2-ylidene]amino]pyridazin- 3-yl]amino]thiazol-4-carboxylate.

A 100 mL single-neck, round-bottom flask was equipped with a PTFE-coated magnetic stir bar and a reflux condenser was installed. It was loaded with 2.6 g of the product from stage A (4.10 mmol, 1 eq.), 1.23 g of NaI (8.2 mmol, 2 eq.) and 20 ml of dry acetone. The reaction mixture was heated to 60°C and stirred at this temperature for 3 days, during which time the reaction reached a state of complete conversion. The reaction mixture was diluted by adding water, and then the precipitated product was collected by filtration, washed with water and then dried under high vacuum to obtain 2.5 g (84%) of the expected product.

Ή NMR (500 MHz, DMSO^) δ 7.82 (d, 1H), 7.61 (s, 1H), 7.47-7.39 (m, 1H), 7.47-7.39 (m, 1H), 7.23 (t, 1H), 5.83 (s, 2H), 4.29 (q, 2H), 3.75 (s, 3H), 3.71 (t, 2H), 3.33 (t, 2H), 3.16 (t, 2H), 2.42 (s, 3H), 2.13 ( quint., 2H), 1.33 (t, 3H), 0.91 (t, 2H), -0.12 (s, 9H); ¹³ C NMR (125 MHz, DMSO^) δ ppm 162.6, 157.3, 156.7, 155.1, 151.6, 140.2, 137.6, 137.1, 135.2, 127.1, 125.4, 123.4, 123.2, 117.5, 111.9, 72.8, 66.7, 60.7, 35.2, 35.2, 27.6, 17.8, 17.8, 14.8, 7.8, -1.0; MSHR-ERI (m/z): [M+H]+, calculated for C ₂₈ H ₃₈ IN ₆ O ₃ S ₂ Si: 725.1255; found: 725.1248.

Synthesis 5h: ethyl 5-[3-[4-[3-tert-butoxycarbonyl-[2-(dimethylamino)ethyl]amino]prop-1-ynyl]-2fluorophenoxy]propyl]-2-[methyl-[5-methyl -6-[(2)-[3-(2-trimethylsilylethoxymethyl)-1,3-benzothiazol-2ylidene]amino]pyridazin-3-yl]amino]thiazol-4-carboxylate.

Step A: tert-butyl N[2-(dimethylamino)ethyl]-L-prop-2-ynylcarbamate.

365 mg of N,N-dimethyl-L-prop-2-ynylethane-1,2-diamine (1.83 mmol, 1 eq.) and 22 mg of N,Ndimethylpyridin-4-amine (0.18 mmol, 0.1 eq.) were mixed in dichloromethane (2.2 ml/mmol), then at room temperature. 600 mg tert-butoxycarbonyl tert-butyl carbonate (2.75 mmol, 1.5 eq.) was added in one portion and the mixture was stirred at ambient temperature for 3 hours. After reaction time, approx. 10 ml DXM and approx. 10 ml conc. NaHCO ₃ , the layers were separated and the organic layer was dried over MgSO ₄ , filtered and concentrated. A yellowish oil was obtained as the crude product: 332 mg (77%) of the expected product.

Ή NMR (400 MHz, DMSOO δ ppm 4.02 (s, 2H), 3.29 (t, 2H), 3.18 (t, 1H), 2.37 (t, 2H), 2.15 (s, 6H), 1.40 (s , 9H); ¹³ C NMR (100 MHz, DMSO<) δ ppm 149.3, 106.7, 79.2, 57.0, 45.3, 43.7, 38.6, 28.0.

Step B: (2-fluoro-4-iodophenoxy)-triisopropylsilane 100 ml oven-dried, single-neck, round-bottom flask was equipped with a PTFE-coated magnetic stir bar. It was loaded with 2.38 g of 2-fluoro-4-iodophenol (10.00 mmol), 1.98 g of K ₂ CO ₃ (20.00 mmol, 2 eq.) and acetonitrile (4 ml/mmol). To the resulting mixture at room temperature. with nearly vigorous stirring, 2.31 g of chloro(triisopropyl)silane (12.00 mmol, 1.2 eq.) was added dropwise. The resulting mixture was stirred at room temperature. for 30 minutes, during which time the reaction reached a state of complete transformation. The reaction mixture was filtered through a celite pad, then concentrated on celite and purified by flash column chromatography using heptane and EtOAc as eluents with

- 62 046336 obtaining 3.70 g (94%) of the target product (94%) in the form of a colorless oil.

1H NMR (500 MHz, DMSO-dg) δ ppm 7.60 (dd, 1H), 7.40 (dm, 1H), 6.82 (dd, 1H), 1.24 (sp, 3H), 1.04 (d, 18H); ¹³ C NMR (125 MHz, DMSOO δ ppm 153.9, 143.8, 134.2, 125.5, 124.1, 83.6, 18.0, 12.5; MSHR-EI (m/z): [M] ⁺ , calculated for Ci ₅ H ₂₄ FIOSi : 394.0625; found: 394.0616.

Step C: tert-butyl N[2-(dimethylamino)ethyl]-N-[3-(3-fluoro-4-triisopropylsilyloxyphenyl)prop-2-ynyl]carbamate.

Using the general Sonogashira reaction procedure, starting from 99 mg of the product from step B (0.25 mmol, 1 eq.) and 73 mg of the product from step A, 90 mg (75%) of the desired product was obtained as a yellow oil.

1H NMR (400 MHz, DMSOO δ ppm 7.09 (dd, 1H), 7.03 (d, 1H), 6.83 (t, 1H), 4.40-4.15 (br., 2H), 3.46 (brt, 2H), 2.50 (t, 2H), 2.26 (s, 6H), 1.47 (s, 9H), 1.25 (m, 3H), 1.07 (d, 18H); ¹³ C NMR (100 MHz, DMSO^) δ ppm. 128.1, 121.6, 119.7, 57.5, 45.7, 44.2, 37.8, 36.8, 28.4, 17.7, 12.7; HRMS-ERI (m/z): [M+H] ⁺ , calculated for C ₂₇ H ₄₆ FN ₂ O ₃ Si: 493.3256; found: 493.3264.

Step D: ethyl 5-[3-[4-[3-[tert-butoxycarbonyl-[2-(dimethylamino)ethyl]amino]prop-1-ynyl]-2fluorophenoxy]propyl]-2-[methyl-[5- methyl 6-[(2)-[3-(2-trimethylsilylethoxymethyl)-1,3-benzothiazol-2ylidene]amino]pyridazin-3-yl]amino]thiazol-4-carboxylate.

Using the general procedure for the alkylation of silyl-protected phenols, starting from 147 mg of the product of synthesis 5g, step B (0.20 mmol, 1 eq.), and 100 mg of the product from step C (0.20 mmol, 1 eq.), 164 mg (87%) were obtained the target product in the form of a brown solid.

1H NMR (500 MHz, DMSO^ ₆ ) δ ppm 7.84 (d, 1H), 7.66 (s, 1H), 7.45 (m, 1H), 7.44 (td, 1H), 7.30 (br, 1H), 7.25 (td, 1H), 7.20 (m, 1H), 7.17 (t, 1H), 5.86 (s, 2H), 4.26 (q, 2H), 4.22 (br, 2H), 4.15 (t, 2H), 3.77 (s, 3H), 3.72 (t, 2H), 3.37 ( m, 2H), 3.27 (t, 2H), 2.48 (m, 2H), 2.45 (s, 3H), 2.22 (br, 6H), 2.12 (quin, 2H), 1.41 (s, 9H), 1.28 (t , 3H), 0.92 (t, 2H), -0.10 (s, 9H); ¹³ C NMR (125 MHz, DMSO^) δ ppm 171.0, 157.6, 156.7, 155.3, 154.7, 151.7, 151.5, 147.6, 141.2, 137.6, 137.1, 135.3, 129.0, 127.2, 125.4, 123.5, 123.1, 119. 2, 117.8, 115.5, 114.7, 111.9, 85.6, 82.1, 79.9, 72.8, 68.7, 66.7, 60.3, 57.0, 45.8, 44.0, 36.9, 35.2, 31.2, 28.5, 23.3, 17.8, 17.5, 14.7, -1.0;

MSHR-ERI (m/z): [M+H] ⁺ , calculated for C ₄₆ H ₆₂ FN ₈ O ₆ S ₂ Si: 933.3982; found: 933.3977.

Synthesis 5i: methyl 5-[3-[4-[3-(tert-butoxycarbonylamino)prop-1-ynyl]-2-fluorophenoxy]propyl]-2[4-[tert-butyl(dimethyl)silyl]oxybutyl-[ 5-methyl-6-[(2)-[3-(2-trimethylsilylethoxymethyl)-1,3benzothiazol-2-ylidene]amino]pyridazin-3-yl]amino]thiazol-4-carboxylate.

Using the general Buchwald reaction procedure II, starting with 780 mg of synthesis product 3g (1.20 mmol, 1 eq.) and 489 mg of synthesis product 4a (1.20 mmol, 1 eq.) as the appropriate halide, 570 mg (47%) of the target product were obtained product.

1H NMR (500 MHz, DMSOO δ ppm 7.83 (dd, 1H), 7.64 (s, 1H), 7.45 (dd, 1H), 7.43 (m, 1H), 7.33 (t, 1H), 7.27 (dd , 1H), 7.26 (m, 1H), 7.18 (m, 1H), 7.15 (t, 1H), 5.85 (s, 2H), 4.41 (t, 2H), 4.14 (t, 2H), 3.93 (d, 2H), 3.76 (s, 3H), 3.71 (t, 2H), 3.61 (t, 2H), 3.25 (t, 2H), 2.44 (s, 3H), 2.12 (m, 2H), 1.74 (m, 2H ), 1.51 (m, 2H), 1.39 (s, 9H), 0.91 (t, 2H), 0.79 (s, 9H), -0.03 (s, 6H), -0.12 (s, 9H); ¹³ C NMR ( 125 MHz, DMSOO δ ppm 162.2, 147.4, 137.6, 129.0, 127.2, 123.4, 123.2, 119.2, 117.6, 115.4, 111.9, 80.8, 72.8, 68.4, 66.7, 62 .5, 51.9, 46.5, 31.1, 30.5, 29.7 , 28.7, 26.1, 23.8, 23.1, 17.9, 17.8, -1.0, -4.9; MSHR-ERI (m/z): [M+H] ⁺ , calculated for C ₅₀ H ₇₁ FN ₇ O ₇ S ₂ Si ₂ : 1020.4373; found: 1020.4373.

Synthesis 5j: ethyl 5-(3-{2-fluoro-4-[3-(methylamino)prop-1-yn-1-yl]phenoxy}propyl)-2-[methyl(5-methyl6-{ [(2Z )-3 -{[2-(trimethylsilyl)ethoxy]methyl}-2,3-dihydro-1,3-benzothiazol-2-ylidene]amino}pyridazin3 -yl)amino]-1,3-thiazol-4-carboxylate .

Step A: ethyl 5-{3-[4-(3-{[(tert-butoxy)carbonyl](methyl)amino}prop-1-yn-1-yl)-2fluorophenoxy]nropyl}-2-[methyl( 5-methyl-6-{[(2Z)-3-{[2-(trimethylsilyl)ethoxy]methyl}-2,3-dihydro-1,3benzothiazol-2-ylidene]amino}pyridazin-3-yl)amino] -1,3-thiazole-4-carboxylate.

To the Synthesis product 5g (1.75 g, 2.41 mmol, 1 equiv.) in dimethylformamide (50 ml) was added the synthesis product 6a (877 mg, 3.14 mmol, 1.3 equiv.) in dimethylformamide (10 ml) and cesium carbonate (2.36 g, 7.24 mmol, 3 eq.), and the mixture was heated at 80°C for 16 hours. The reaction mixture was concentrated in vacuo, then partitioned between ethyl acetate and brine, and the organic phase was dried (magnesium sulfate) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 40 g RediSep™ silica gel cartridge) eluting with a gradient of 0-50% ethyl acetate in isoheptane gave the title product as a yellow oil (1.75 g, 2 mmol, 83%).

LC/MS (C ₄₃ H ₅₄ FN ₇ O ₆ SiS ₂ ) 876 [M+H] ⁺ ; RT 1.46 (ZHMS-U-V2).

1H NMR (400 MHz, DMSOO δ 7.83 (dd, 1H), 7.65 (d, J=1.1 Hz, 1H), 7.49-7.39 (m, 2H), 7.35-7.28 (m, 1H), 7.27-7.12 (m , 3H), 5.86 (s, 2H), 4.25 (q, J=7.1 Hz, 2H), 4.19 (s, 2H), 4.14 (t, J=6.1 Hz, 2H), 3.77 (s, 3H), 3.76 - 3.68 (m, 2H), 3.26 (t, J=7.7 Hz, 2H), 2.84 (s, 3H), 2.45 (s, 3H), 2.19-2.05 (m, 1H), 1.41 (s, 9H), 1.30 (t, 3H), 0.97-0.88 (m, 2H), -0.12 (s, 9H).

Step B: ethyl 5-(3-{2-fluoro-4-[3-(methylamino)prop-1-yn-1-yl]phenoxy}propyl)-2-[methyl(5-methyl6-{ [(2Z )-3-{ [2-(trimethylsilyl)ethoxy]methyl}-2,3-dihydro-1,3-benzothiazol-2-ylidene]amino}pyridazin3 -yl)amino]-1,3-thiazol-4-carboxylate .

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Trifluoroacetic acid (20 ml) was added to a stirred solution of the product from step A (1.5 g, 1.71 mmol, 1 eq.) in dichloromethane (60 ml), and the mixture was stirred at ambient temperature for 5 hours. The reaction mixture was diluted with dichloromethane, cooled to 0°C and made alkaline by adding 2N. aqueous sodium hydroxide solution. The organic phase was dried (magnesium sulfate) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 40 g RediSep™ silica gel cartridge) eluting with a gradient of 0-10% methanol in dichloromethane gave the title product as a yellow gum (329 mg, 0.42 mmol, 25%).

LC/MS (C ₃₈ H ₄₆ FN ₇ O ₄ SiS ₂ ) 776 [M+H]+; RT 2.58 (LCMS-VC).

1H NMR (400 MHz, DMSO<) δ 7.84 (dd, 1H), 7.67 (d, J=1.0 Hz, 1H), 7.49-7.40 (m, 2H), 7.31-7.22 (m, 2H), 7.21-7.11 (m, 2H), 5.86 (s, 2H), 4.26 (q, J=7.1 Hz, 2H), 4.15 (t, J=6.1 Hz, 2H), 3.76 (s, 3H), 3.76-3.67 (m, 2H), 3.45 (s, 2H), 3.33-3.22 (m, 2H), 2.46 (d, J=1.0 Hz, 3H), 2.30 (s, 3H), 2.18-2.06 (m, 2H), 1.29 (t , J=7.1 Hz, 3H), 0.97-0.88 (m, 2H), -0.11 (s, 9H).

Synthesis 5k: ethyl 2-[methyl(5-methyl-6-{[(22)-3-{[2-(trimethylsilyl)ethoxy]methyl}-2,3-dihydro-1,3benzothiazol-2-ylidene]amino }pyridazin-3-yl)amino]-5-(piperidin-4-yl)-1,3-thiazol-4-carboxylate.

Step A: bromine({1-[(tert-butoxy)carbonyl]piperidin-4-yl})zinc.

A 50 mL Schlenk flask equipped with a magnetic stir bar was charged with zinc (1.96 g, 30 mmol, 1.5 eq.) and lithium chloride (1.27 g, 30 mmol, 1.5 eq.), and the mixture was heated with a heat gun under high vacuum (< 1 mbar) for 10 minutes with gentle stirring. Still under high vacuum, the mixture was allowed to cool to ambient temperature and then transferred to a Schlenk line, which was filled with nitrogen. Tetrahydrofuran (15 ml) was added followed by 1,2-dibromoethane (0.06 ml, 0.75 mmol, 0.04 eq) and the mixture was heated to 60°C for 10 min. Heating of the reaction mixture was then stopped and trimethylchlorosilane (0.02 mL, 0.15 mmol, 0.01 eq.) was added, followed by the addition of iodine (0.5 M in tetrahydrofuran; 0.15 mL, 0.175 mmol, 0.05 eq.). The mixture was then heated at 60°C for 10 minutes and then cooled to <35°C. A solution of tert-butyl 4bromopiperidine-1-carboxylate (5.28 g, 20 mmol, 1 equiv) in tetrahydrofuran (10 ml) was added over 3 min and the mixture was stirred at 50°C for 6 h and then at ambient temperature at during the night. The reaction mixture was allowed to cool to ambient temperature and then cannulated through a filter (cotton wadding/celite/cotton wadding) under low vacuum in a dry 25 ml Schlenk flask to obtain the desired product as a 0.5 M solution (as determined by titration with a 0.5 M solution iodine), which was used without further characterization.

Step B: tert-butyl 4-[4-(ethoxycarbonyl)-2-[methyl(5-methyl-6-{[(22)-3-{[2(trimethylsilyl)ethoxy]methyl}-2,3-dihydro -1,3-benzothiazol-2-ylidene]amino}pyridazin-3-yl)amino]1,3-thiazol-5-yl]piperidine-1-carboxylate.

Synthesis product 11b (500 mg, 0.79 mmol, 1 eq.) and copper iodide ([) (300 mg, 1.57 mmol, 2 eq.) were loaded into an oven-dried flask, and the mixture was evacuated and purged with nitrogen (x3). Dimethylacetamide (15 ml) was added resulting in a brown suspension which was allowed to stir for 5 minutes. The product from Step A (0.5 M in tetrahydrofuran; 9.44 mL, 4.72 mmol, 6 eq.) was added and the mixture was stirred at ambient temperature overnight. The reaction mixture was quenched by adding saturated aqueous ammonium chloride (25 ml), and then further diluted 1:1 with water/saturated aqueous ammonium chloride (150 ml). Organics were extracted with dichloromethane (x3) and the combined organic extracts were washed with brine, dried (magnesium sulfate) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 40 g RediSep™ silica gel cartridge) eluting with a gradient of 0-40% ethyl acetate in isoheptane gave the title product as a pale yellow foam (429 mg, 0.58 mmol, 74% ).

LC/MS (C ₃₅ H ₄₉ N ₇ O ₅ SiS ₂ ) 740 [M+H]+; RT 1.40 (ZHMS-U-V2).

1H NMR (400 MHz, CDCl ₃ ) δ 7.63 (dt, J=7.7, 1.0 Hz, 1H), 7.46-7.34 (m, 2H), 7.31 (d, J=1.2 Hz, 1H), 7.22 (ddd, J =7.7, 5.8, 2.6 Hz, 1H), 5.85 (s, 2H), 4.40 (q, J=7.1 Hz, 2H), 4.26 (s, 2H), 3.90 (ddd, J=12.1, 8.5, 3.7 Hz, 1H), 3.85 (s, 3H), 3.80-3.69 (m, 2H), 2.85 (s, 2H), 2.47 (d, J=1.0 Hz, 3H), 2.04 (d, J=12.8 Hz, 2H), 1.71 (q, J=12.1 Hz, 2H), 1.50 (s, 9H), 1.47 (s, 0H), 1.44 (t, J=7.1 Hz, 3H), 1.02-0.93 (m, 2H), -0.07 ( s, 9H).

Step C: ethyl 2-[methyl(5-methyl-6-{[(22)-3-{[2-(trimethylsilyl)ethoxy]methyl}-2,3-dihydro-1,3benzothiazol-2-ylidene]amino }pyridazin-3-yl)amino]-5-(piperidin-4-yl)-1,3-thiazol-4-carboxylate.

Trifluoroacetic acid (0.5 mL, 6.71 mmol, 49.7 eq.) was added slowly to a cooled solution of the product from Step B (100 mg, 0.14 mmol, 1 eq.) in dichloromethane (6 mL), and the mixture was stirred at 0°C for 1.5 h. The reaction mixture was quenched by adding saturated aqueous sodium bicarbonate, extracted with dichloromethane, and the organic extract was washed with brine, dried (magnesium sulfate) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 12 g RediSep™ silica gel cartridge) eluting with a gradient of 0-10% methanol in dichloromethane gave the title product as an off-white glassy solid (59 mg, 0.09 mmol, 68 %).

- 64 046336

LC/MS (C ₃₀ H ₄₁ N ₇ O ₃ SiS ₂ ) 640 [M+H]+; RT 1.23 (LCMS-V-Bl).

1H NMR (400 MHz, DMSO^ ₆ ) δ 7.85 (dd, J=7.6, 1.1 Hz, 1H), 7.67 (d, J=1.1 Hz, 1H), 7.51-7.40 (m, 2H), 7.25 (ddd, J=8.3, 6.9, 1.5 Hz, 1H), 5.87 (s, 2H), 4.29 (q, J=7.1 Hz, 2H), 3.90-3.61 (m, 6H), 3.12 (d, J=12.2 Hz, 2H ), 2.68 (td, J=12.5, 2.3 Hz, 3H), 2.46 (d, J=1.0 Hz, 3H), 2.00-1.89 (m, 2H), 1.71-1.54 (m, 2H), 1.32 (t, J=7.1 Hz, 3H), 0.92 (dd, J=8.6, 7.4 Hz, 2H), -0.11 (s, 9H).

Synthesis 51: ethyl 5-(azetidin-3-yl)-2-[methyl(5-methyl-6-{[(22)-3-{[2-(trimethylsilyl)ethoxy]methyl}2,3-dihydro- 1,3-benzothiazol-2-ylidene]amino}pyridazin-3-yl)amino] - 1,3-thiazol-4-carboxylate.

Step A: ethyl 5-{1-[(tert-butoxy)carbonyl|azetidin-3-yl}-2-[methyl(5-methyl-6-{[(22)-3-{[2(trimethylsilyl)ethoxy ]methyl}-2,3-dihydro-1,3-benzothiazol-2-ylidene]amino}pyridazin-3-yl)amino]1,3-thiazol-4-carboxylate.

To a solution of synthesis product 4a (401 mg, 0.98 mmol, 2 equiv.) in 1,4-dioxane (5 ml) was added synthesis product 3zb (168 mg, 0.49 mmol, 1 equiv.), N,N-diisopropylethylamine (0.24 ml , 1.48 mmol, 3 eq.), cesium carbonate (481 mg, 1.48 mmol, 3 eq.) and xantphos (56.9 mg, 0.1 mmol, 0.2 eq.), and the mixture was bubbled with nitrogen (10 min). Tris(dibenzylideneacetone)dipalladium(0) (45.1 mg, 0.05 mmol, 0.1 equiv) was added and the mixture was heated at 120°C for 2 h under microwave irradiation. The reaction mixture was diluted with dichloromethane, then washed with water followed by brine, dried (magnesium sulfate) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 24 g RediSep™ silica gel cartridge) eluting with a gradient of 0-100% ethyl acetate in isoheptane gave the title product as a brown solid (99 mg, 0.14 mmol, 28%).

LC/MS (C ₃₃ H ₄₅ N ₇ O ₅ SiS ₂ ) 712 [M+H]+; RT 1.52 (LCMS-^B1).

1H NMR (400 MHz, DMSO<) δ 7.90-7.81 (m, 1H), 7.70 (d, J=1.1 Hz, 1H), 7.52-7.39 (m, 2H), 7.29-7.21 (m, 1H), 5.87 (s, 2H), 4.59 (tt, J=8.6, 6.0 Hz, 1H), 4.37 (t, J=8.6 Hz, 2H), 4.29 (q, J=7.1 Hz, 2H), 3.89 (t, J= 7.3 Hz, 2H), 3.79 (s, 3H), 3.76-3.68 (m, 2H), 2.46 (d, J=1.0 Hz, 3H), 1.43 (s, 9H), 1.32 (t, J=7.1 Hz, 3H), 0.99 - 0.83 (m, 2H), -0.11 (s, 9H).

Step B: ethyl 5-(azetidin-3-yl)-2-[methyl(5-methyl-6-{[(22)-3-{[2-(trimethylsilyl)ethoxy]methyl}2,3-dihydro- 1,3-benzothiazol-2-ylidene]amino}pyridazin-3-yl)amino] - 1,3-thiazol-4-carboxylate.

To a solution of the product from step A (78 mg, 0.11 mmol, 1 eq.) in dichloromethane (6 mL) cooled in an ice bath, trifluoroacetic acid (0.5 mL, 6.71 mmol, 61.2 eq.) was slowly added and the mixture was stirred at 0°C for 5 hours. The reaction mixture was quenched by adding saturated aqueous sodium bicarbonate, then extracted with dichloromethane, washed with brine, dried (magnesium sulfate) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 12 g RediSep™ silica gel cartridge) eluting with a gradient of 0-10% methanol in dichloromethane provided the title product as a cream solid (29.6 mg, 0.05 mmol, 44%) .

LC/MS (C ₂₈ H ₃₇ N ₇ O ₃ SiS ₂ ) 612 [M+H]+; RT 1.18 (LCMS-^B1).

1H NMR (400 MHz, DMSO-de) δ 7.83 (d, J=7.7 Hz, 1H), 7.71 (s, 1H), 7.52 -7.40 (m, 2H), 7.30-7.23 (m, 1H), 5.87 ( s, 2H), 4.76 (p, J=8.2 Hz, 1H), 4.29 (q, J=7.1 Hz, 2H), 4.17 (t, J=9.0 Hz, 2H), 3.91 (t, J=8.5 Hz, 2H), 3.79 (s, 3H), 3.75-3.66 (m, 2H), 2.47 (s, 3H), 1.32 (t, J=7.1 Hz, 3H), 0.98-0.87 (m, 2H), -0.11 ( s, 9H).

Synthesis 5t: ethyl 5-[(1E)-3-hydroxy-2-methylprop-1-en-1-yl]-2-[methyl(5-methyl-6-{[(22)-3-{[2 (trimethylsilyl)ethoxy]methyl}-2,3-dihydro-1,3-benzothiazol-2-ylidene]amino}pyridazin-3-yl)amino]1,3-thiazol-4-carboxylate.

Step A: tert-butyldimethyl(prop-2-yn-1-yloxy)silane.

To a cooled solution of propargyl alcohol (5.21 ml, 89.2 mmol, 1 eq.) in dichloromethane (250 ml) was added imidazole (8.84 ml, 134 mmol, 1.5 eq.) and tert-butyldimethylsilyl chloride (20.2 g, 134 mmol, 1.5 eq.) , and the mixture was stirred at 0°C for 90 minutes. The reaction mixture was partitioned between dichloromethane and saturated aqueous ammonium chloride, the layers were separated (PTFE phase separator), and the organic phase was concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 220 g RediSep™ silica gel cartridge) eluting with a gradient of 0-10% ethyl acetate in isoheptane gave the title product as a clear oil (4.07 g, 23.9 mmol, 27%).

1H NMR (400 MHz, DMSO-de) δ 4.28 (d, J=2.4 Hz, 2H), 3.37 (t, J=2.4 Hz, 1H), 0.86 (s, 9H), 0.08 (s, 6H).

Step B: tert-butyldimethyl{[(2E)-2-methyl-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)prop2-en-1-yl]oxy }silane.

Copper(1) chloride (237 mg, 2.39 mmol, 0.1 equiv.), xanthos (1.38 g, 2.39 mmol, 0.1 equiv.), bis(pinacolato)dibor (6.67 g, 26.3 mmol, 1.1) was loaded into a flask dried in an oven. eq.) and tetrahydrofuran (17 ml), and the mixture was bubbled with nitrogen (10 min). A solution of sodium 2-methylpropan-2-olate (2.53 g, 26.3 mmol, 1.1 eq) in tetrahydrofuran (14 ml) was added and the mixture was stirred for 5 min. A solution of the product from step A (4.07 g, 23.9 mmol, 1 eq) in tetrahydrofuran (14 ml) was added followed by methyl iodide (5.95 ml, 95.6 mmol, 4 eq) and the mixture was stirred at ambient temperature overnight. The reaction mixture was partitioned between ethyl acetate and water, and

- 65 046336 the ganic phase was dried (magnesium sulfate) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 80 g RediSep™ silica gel cartridge) eluting with a gradient of 0-5% ethyl acetate in isoheptane gave the title product as a clear oil (616 mg, 1.97 mmol, 8%).

1H NMR (400 MHz, DMSO^b) δ 5.34-5.30 (m, 1H), 4.04-3.97 (m, 2H), 1.83 (s, 3H), 1.23-1.16 (m, 12H), 0.88 (s, 9H ), 0.03 (s, 6H).

Step C: ethyl 5-[(1E)-3-[(tert-butyldimethylsilyl)oxy]-2-methylprop-1-en-1-yl]-2-[methyl(5methyl-6-{[(22)- 3-{[2-(trimethylsilyl)ethoxy]methyl}-2,3-dihydro-1,3-benzothiazol-2ylidene] amino } pyridazin-3 -yl) amino]-1,3-thiazol-4-carboxylate.

To a solution of synthesis product 11e (224 mg, 0.33 mmol, 1 eq.) in tetrahydrofuran (7.5 ml) was added the product from step B (123 mg, 0.39 mmol, 1.2 eq.) followed by the addition of water (2.5 ml) and potassium carbonate ( 136 mg, 0.98 mmol, 3 eq.). The mixture was bubbled with nitrogen (10 min), after which [1,1'bis(diphenylphosphino)ferrocene]dichloropalladium(II) (24 mg, 0.03 mmol, 0.1 eq.) was added and the mixture was heated at 100°C for 1 h under microwave irradiation. The reaction mixture was partitioned between ethyl acetate and brine, and the organic phase was dried (magnesium sulfate) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 12 g RediSep™ silica gel cartridge) eluting with a gradient of 0-40% ethyl acetate in isoheptane provided the title product as a white solid (192 mg, 0.26 mmol, 79%) .

LC/MS (C ₃₅ H ₅₂ N ₆ O ₄ Si ₂ S ₂ ) 741 [M+H]+; RT 1.58 (ZHMS-U-V2).

1H NMR (400 MHz, DMSO<) δ 7.86-7.79 (m, 1H), 7.70 (d, J=1.1 Hz, 1H), 7.51-7.40 (m, 2H), 7.29-7.20 (m, 2H), 5.86 (s, 2H), 4.28 (q, 2H), 4.22 (s, 2H), 3.79 (s, 3H), 3.76-3.67 (m, 2H), 2.46 (s, 3H), 1.93 (s, 3H), 1.30 (t, 3H), 0.93 (s, 9H), 0.91-0.83 (m, 2H), 0.11 (s, 6H), -0.12 (s, 9H).

Step D: ethyl 5-[(1E)-3-hydroxy-2-methylprop-1-en-1-yl]-2-[methyl(5-methyl-6-{[(22)-3-{[2 (trimethylsilyl)ethoxy]methyl}-2,3-dihydro-1,3-benzothiazol-2-ylidene]amino}pyridazin-3-yl)amino]1,3-thiazol-4-carboxylate.

To a solution of the product from step C (192 mg, 0.26 mmol, 1 eq.) in tetrahydrofuran (6 ml) was added tetrabutylammonium fluoride (1 M in tetrahydrofuran; 0.39 ml, 0.39 mmol, 1.5 eq.) and the mixture was stirred at ambient temperature for 2 hours. The reaction mixture was partitioned between ethyl acetate and water, and the organic phase was dried (magnesium sulfate) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 4 g RediSep™ silica gel cartridge) eluting with a gradient of 0-60% ethyl acetate in isoheptane provided the title product as a yellow solid (160 mg, 0.26 mmol, 99%) .

LC/MS (C2 ₉ H38N6O4SiS2) 627 [M+H]+; RT 2.71 (LCMS-VC).

1H NMR (400 MHz, DMSO<) δ 7.86-7.80 (m, 1H), 7.70 (d, J=1.1 Hz, 1H), 7.49-7.39 (m, 2H), 7.28-7.21 (m, 2H), 5.86 (s, 2H), 5.12 (t, J=5.7 Hz, 1H), 4.29 (q, J=7.1 Hz, 2H), 4.09-3.98 (m, 2H), 3.81 (s, 3H), 3.77-3.67 ( m, 2H), 2.46 (d, J=6.8 Hz, 3H), 1.94 (s, 3H), 1.31 (t, J=7.1 Hz, 3H), 0.97-0.89 (m, 2H), -0.12 (s, 9H).

Synthesis 6a: tert-Butyl N[3-(3-fluoro-4-hydroxyphenyl)prop-2-ynyl]-N-methylcarbamate.

Using the general Sonogashira reaction procedure, starting with 10.00 g of 2-fluoro-4iodophenol (42.0 mmol, 1 eq.) as the appropriate phenol and 10.67 g of tert-butyl N-methyl-N-prop-2ynylcarbamate (63.1 mmol, 1.5 eq.) as an alkyne reagent, 10.8 g (92%) of the target product was obtained.

1H NMR (500 MHz, DMSO<) δ ppm 10.32 (s, 1H), 7.22 (brd, 1H), 7.08 (dm, 1H), 6.92 (dd, 1H), 4.21 (s, 2H), 2.85 (s, 3H), 1.41 (s, 9H); ¹³ C NMR (125 MHz, DMSO-d^ δ ppm 150.8, 146.4, 129.0, 119.6, 118.4, 113.2, 84.4, 82.7, 38.5, 33.8, 28.5; HRMS-ERI (m/z): [M- C4H8+H]+, calculated for C _n H _n FNO ₃ : 224.0717; found: 224.0720.

Synthesis 6b: 4-[3-(dimethylamino)prop-1-ynyl]-2-fluorophenol.

Using the general Sonogashira reaction procedure, starting with 10.00 g of 2-fluoro-4iodophenol (42.0 mmol, 1 eq.) as the appropriate phenol and 5.24 g of KH-dimethylprop-2-yne-1-amine (63 mmol, 1.5 eq.) as an alkyne reagent, 7.30 g (90%) of the target product was obtained.

1H NMR (500 MHz, DMSO^6) δ ppm 7.20 (dd, 1H), 7.07 (dm, 1H), 6.91 (m, 1H), 3.39 (m, 2H), 2.21 (m, 3H); ¹³ C NMR (125 MHz, DMSO<) δ ppm 150.9, 146.2, 128.9, 119.5, 118.4, 113.6, 84.5, 84.2, 48.2, 44.3; MSHR-ERI (m/z): [M+H]+, calculated for C _n H ₁₃ FNO: 194.0976; found: 194.0981.

Synthesis of 6c: 2-fluoro-4-(3-pyrrolidin-1-ylprop-1-ynyl)phenol.

Step A: 3-(3-fluoro-4-triisopropylsilyloxyphenyl)prop-2-yn-1-ol.

4.76 g of 2-fluoro-4-iodophenol (20 mmol, 1 eq.) and 3.96 g of K2CO3 (40 mmol, 2 eq. ), and then 100 ml of dry MeCN was added. To the resulting mixture at room temperature. With nearly vigorous stirring, 5.13 mL of TIPSCl (4.62 g, 24 mmol, 1.2 eq.) was added dropwise. The resulting mixture was stirred at room temperature for 30 minutes, during which time the reaction reached a state of complete conversion. The reaction mixture was filtered through a celite pad to remove solid particles, then 2.33 ml of prop-2-in-1 was added to the filtrate

- 66 046336 ol (2.24 g, 40 mmol, 2 eq.) and 20 ml DIPA, and a nitrogen atmosphere was created through the gas inlet. After adding 702 mg Pd(PPh ₃ ) ₂ Cl ₂ (1 mmol, 0.05 eq.) and 190 mg CuI (1 mmol, 0.05 eq.), the resulting mixture was stirred at room temperature for 30 min, during which time the reaction reached state of complete transformation. Celite was added to the reaction mixture, and volatiles were removed under reduced pressure. The crude product was then purified by flash column chromatography using heptane and EtOAc as eluents to give 5.2 g (81%) of the title product as a brown oil.

^1H NMR (500 MHz, DMSOHO δ ppm 7.26 (dd, 1H), 7.14 (dm, 1H), 6.96 (t, 1H), 4.27 (d, 2H), 1.23 (m, 3H), 1.03 ( d, 18H); ¹³ C NMR (125 MHz, DMSO^) δ ppm 153.3, 144.1, 128.8, 122.3, 119.6, 116.5, 89.8, 82.8, 49.8, 17.9, 12.5; HRMS-ER (m/z) : [M+H]+, calculated for C ₁₈ H ₂₈ FO ₂ Si: 323.1837; found: 323.1832.

Step B: [2-fluoro-4-(3-pyrrolidin-1-ylprop-1-ynyl)phenoxy]triisopropylsilane.

Using a general alkylation procedure with an in situ formed iodine-containing compound, starting with 322 mg of the product from Step A (1 mmol, 1 eq.) as the appropriate alcohol and 355 mg of pyrrolidine (5 mmol, 5 eq.) gave 130 mg (34%) target product.

1H NMR (500 MHz, DMSO^ ₆ ) δ ppm 7.29 (dd, 1H), 7.14 (dm, 1H), 6.97 (t, 1H), 3.56 (s, 2H), 2.55 (m, 4H), 1.71 (m, 4H), 1.25 (m, 3H), 1.05 (d, 18H); ¹³ C NMR (125 MHz, DMSOL) δ ppm 153.2, 144.0, 129.0, 122.3, 119.8, 116.6, 86.2, 83.2, 52.3, 43.3, 23.8, 18.0, 12.5; MSHR-ERI (m/z): [M+H]+, calculated for C ₂₂ H ₃₅ FNOSi: 376.2466; found: 376.2456.

Step C: 2-fluoro-4-(3-pyrrolidin-1-ylprop-1-ynyl)phenol.

A 4 ml oven-dried vessel equipped with a PTFE-coated magnetic stirrer arm was charged with 83 mg of the product from step B (0.221 mmol, 1 eq.) and 1.1 ml of dry THF, then at room temperature. 265 μl of TBAF (1 M in THF, 225.9 mg, 0.2652 mmol, 1.2 eq.) was added dropwise. The resulting mixture was stirred at room temperature. for 15 minutes, during which the reaction reached a state of complete transformation. The reaction mixture was quenched by adding 200 μl of conc. NH4CI solution, and then celite was added to the reaction mixture and the volatiles were removed under reduced pressure. The crude product was then purified by flash column chromatography using DCM and MeOH (1.2% NH3) as eluents to obtain 43 mg (88%) of the title product.

1H NMR (500 MHz, DMSOHO δ ppm 10.30 (brs, 1H), 7.17 (dd, 1H), 7.04 (dm, 1H), 6.88 (t, 1H), 3.53 (s, 2H), 2.54 (m ^- ERI (m/z): [M+H] ⁺ , calculated for C^FNO: 220.1132; found: 220.1129.

Synthesis of 6d: 2-fluoro-4-[3-(1-piperidyl)prop-1-ynyl]phenol.

Step A: [2-fluoro-4-[3-(1-piperidyl)prop-1-ynyl]phenoxy]triisopropylsilane.

Using a general alkylation procedure with an in situ formed iodine-containing compound, starting from 322 mg of the product of synthesis 6c, step A (1 mmol, 1 eq.) as a suitable alcohol and 425 mg of piperidine (5 mmol, 5 eq.), gave 250 mg ( 64%) of the target product.

1H NMR (500 MHz, DMSOO δ ppm 7.3 (dd, 1H), 7.15 (dm, 1H), 6.98 (t, 1H), 3.42 (s, 2H), 2.46 (br., 4H), 1.51 ( m, 4H), 1.37 (br., 2H), 1.26 (m, 3H), 1.05 (d, 18H); ¹³ C NMR (125 MHz, DMSO^) δ ppm 153.2, 144.0, 129.0, 122.3, 119.8, 116.6, 85.9, 83.8, 53.1, 48.0, 25.9, 24.0, 18.0, 12.5; MSHR-ERI (m/z): [M+H]+, calculated for C ₂₃ H ₃₇ FNOSi: 390.2622; found: 390.2608.

Step B: 2-fluoro-4-[3-(1-piperidyl)prop-1-ynyl]phenol.

A 4 mL oven-dried vessel equipped with a PTFE-coated magnetic stir bar was charged with 272 mg of the product from step A (0.69 mmol, 1 eq.), dissolved in 3.5 mL of dry THF, and then at room temperature. 698 μl of TBAF (1 M in THF, 0.69 mmol, 1 eq.) was added dropwise. The resulting mixture was stirred at room temperature. for 15 minutes, during which the reaction reached a state of complete transformation. The reaction mixture was quenched by adding 200 μl of conc. NH ₄ Cl solution, and then celite was added to the reaction mixture, and the volatiles were removed under reduced pressure. The crude product was then purified by flash column chromatography using DCM and MeOH (1.2% NH3) as eluents to obtain 143 mg (87%) of the desired product.

1H NMR (500 MHz, DMSOO δ ppm 10.27 (s, 1H), 7.20 (dd, 1H), 7.07 (dm, 1H), 6.90 (t, 1H), 3.40 (s, 2H), 2.44 (br , 4H), 1.51 (m, 4H), 1.37 (br, 2H); ¹³ C NMR (125 MHz, DMSOO δ ppm 150.9, 146.1, 128.9, 119.5, 118.4, 113.7, 84.7, 84.2, 53.1, 48.0 , 25.9, 24.1; MSHR-ERI (m/z): [M+H]+, calculated for C14H17FNO: 234.1289; found: 234.1292.

Synthesis 6e: 2-Fluoro-4-(3-morpholinoprop-1-ynyl)phenol.

Step A: [2-fluoro-4-(3-morpholinoprop-1-ynyl)phenoxy]triisopropylsilane.

Using a general alkylation procedure with an in situ formed iodine-containing compound, starting from 322 mg of the product of synthesis 6c, step A (1 mmol, 1 eq.), as a suitable alcohol and 435 mg of morpholine (5 mmol, 5 eq.), gave 160 mg ( 41%) of the target product.

1H NMR (500 MHz, DMSOO δ ppm 7.32 (dd, 1H), 7.16 (dd, 1H), 6.98 (t, 1H), 3.6 (t, 4H), 3.47 (s, 2H), 2.49 (t , 4H), 1.25 (m, 3H), 1.05 (d, 18H); ¹³ C NMR (125 MHz, DMSO<) δ ppm 153.3, 144.1, 129.0, 122.3, 119.8, 116.4, 85.3, 84.1, 66.6 , 52.3, 47.5, 18.0, 12.5; MSVR-ERI (m/z): [M+H]+, calculated for

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C ₂₂ H ₃₅ FNO2Si: 392.2415; found: 392.2408.

Step B: 2-fluoro-4-(3-morpholinoprop-1-ynyl)phenol.

A 4 mL oven-dried vessel equipped with a PTFE-coated magnetic stir bar was charged with 220 mg of the product from step A (0.56 mmol, 1 eq.), dissolved in 3.0 mL of dry THF, and then at room temperature. 525 μl of TBAF (1 M in THF, 0.52 mmol, 1 eq.) was added dropwise. The resulting mixture was stirred at room temperature. for 15 minutes, during which the reaction reached a state of complete transformation. The reaction mixture was quenched by adding 200 μl of conc. NH ₄ Cl solution, and then celite was added to the reaction mixture, and the volatiles were removed under reduced pressure. The crude product was purified by flash column chromatography using DCM and MeOH (1.2% NH3) as eluents to give 120 mg (91%) of the title product.

1H NMR (400 MHz, DMSO<) δ ppm 10.28 (s, 1H), 7.22 (dd, 1H), 7.08 (dm, 1H), 6.91 (dd, 1H), 3.60 (m, 4H), 3.45 (s, 2H), 2.48 (m, 4H); ¹³ C NMR (125 MHz, DMSO^) δ ppm 150.8, 146.2, 128.9, 119.5, 118.4, 113.6, 84.6, 84.1, 66.5, 52.2, 47.5; MSHR-ERI (m/z): [M+H]+, calculated for Ci ₃ Hi ₃ FNO2: 236.1081; found: 236.1082.

Synthesis of 6f: 4-[3-(dimethylamino)but-1-ynyl]-2-fluorophenol.

Step A: 4-(3-fluoro-4-triisopropylsilyloxyphenyl)but-3-yn-2-ol.

A 500 mL oven-dried, single-neck, round-bottom flask was equipped with a PTFE-coated magnetic stir bar. 4.76 g of 2-fluoro-4-iodophenol (20 mmol, 1 eq.) and 3.96 g of K2CO3 (40 mmol, 2 eq.) were loaded into it, and then 100 ml of dry MeCN was added. To the resulting mixture at room temperature. With nearly vigorous stirring, 5.13 mL of TIPSCl (4.62 g, 24 mmol, 1.2 eq.) was added dropwise. The resulting mixture was stirred at room temperature for 30 minutes, during which time the reaction reached a state of complete conversion. The reaction mixture was filtered through a pad of Celite to remove solids, then 3.10 mL of but-3-yn-2-ol (2.81 g, 40 mmol, 2 eq.) and 20 mL of DIPA were added to the filtrate, and a nitrogen atmosphere was created through the gas inlet. After adding 702 mg Pd(PPh ₃ ) ₂ Cl ₂ (1 mmol, 0.05 eq.) and 190 mg CuI (1 mmol, 0.05 eq.), the resulting mixture was stirred at room temperature for 30 min, during which time the reaction reached state of complete transformation. Celite was added to the reaction mixture, and volatiles were removed under reduced pressure. The crude product was then purified by flash column chromatography using heptane and EtOAc as eluants to give 6.2 g (92%) of the title product as a yellow oil.

1H NMR (400 MHz, DMSO<) δ ppm 7.26 (dd, 1H), 7.12 (dm, 1H), 6.98 (t, 1H), 5.44 (d, 1H), 4.55 (m, 1H), 1.36 (d, 3H), 1.24 (sp, 1H), 1.05 (d, 18H); ¹³ C NMR (100 MHz, DMSO^) δ ppm 153.2, 144.1, 128.8, 122.3, 119.6, 116.5, 93.4, 81.4, 57.1, 25.0, 18.0, 12.5; MSHR-ERI (m/z): [M+H]+, calculated for C ₁₉ H ₃₀ FO ₂ Si: 337.1994; found: 337.1994.

Step B: 4-(3-fluoro-4-triisopropylsilnoxyphenyl)-^Ndimethyl-but-3-yn-2-amine.

Using a general alkylation procedure with an in situ formed iodine-containing compound, starting from 644 mg of the product from step A (2 mmol, 1 eq.) as the appropriate alcohol and 5 ml of Nmethylmethanamine (10 mmol, 5 eq., 2 M solution in MeOH) gave 360 mg (50%) of the target product.

1H NMR (500 MHz, DMSO<) δ ppm 7.28 (dd, 1H), 7.14 (dm, 1H), 6.97 (t, 1H), 3.67 (q, 1H), 2.19 (s, 6H), 1.27 (d, 3H), 1.25 (m, 3H), 1.05 (d, 18H); ¹³ C NMR (500 MHz, DMSOL) δ ppm 153.1, 144.0, 129.0, 122.3, 119.8, 116.6, 88.2, 84.1, 52.3, 41.3, 20.1, 18.0, 12.5; MSHR-ERI (m/z): [M+H]+, calculated for C ₂₁ H ₃₅ FNOSi: 364.2466; found: 364.2470.

Step C: 4-[3-(dimethylamino)but-1-ynyl]-2-fluorophenol.

A 4 mL oven-dried vessel equipped with a PTFE-coated magnetic stir bar was charged with 200 mg of the product from step B (0.55 mmol, 1 eq.) dissolved in 3.0 mL of dry THF, and then at room temperature. 660 μl of TBAF (1 M in THF, 0.66 mmol, 1.1 eq.) was added dropwise. The resulting mixture was stirred at room temperature. for 15 minutes, during which the reaction reached a state of complete transformation. The reaction mixture was quenched by adding 200 μl of conc. NH ₄ Cl solution, and then celite was added to the reaction mixture, and the volatiles were removed under reduced pressure. The crude product was then purified by flash column chromatography using DCM and MeOH (1.2% NH ₃ ) as eluents to obtain 80 mg (70%) of the title product.

Synthesis of 6g and 6h: 2-fluoro-4-(3-morpholinobut-1-ynyl)phenol (enantiomers 1 and 2).

Step A: [2-fluoro-4-(3-morpholnobut-1-ynyl)phenoxy]triisopropylsilane.

Using a general alkylation procedure with an in situ formed iodine-containing compound, starting with 644 mg of synthesis product 6f, step A (2 mmol, 1 eq.) as a suitable alcohol and 5 ml of morpholine (10 mmol, 5 eq., 2 M solution in MeOH ) obtained 370 mg (45%) of the target product.

1H NMR (500 MHz, DMSO<) δ ppm 7.29 (dd, 1H), 7.15 (dm, 1H), 6.98 (t, 1H), 3.69 (q, 1H), 3.60 (m, 4H), 2.6/2.45 (m+m, 4H), 1.30 (d, 3H), 1.25 (m, 3H), 1.05 (d, 18H); ¹³ C NMR (500 MHz, DMSOL) δ ppm 153.2, 144.0, 129.0, 122.3, 119.8, 116.5, 88.5, 84.1, 66.7, 52.1, 49.5, 19.3, 18.0, 12.5; MSHR-ERI (m/z): [M+H] ⁺ , calculated for C ₂₃ H ₃₇ FNO ₂ Si: 406.2572; found: 406.2579.

Step B: 2-fluoro-4-(3-morpholinobut-1-ynyl)phenol.

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A 4 mL oven-dried vessel equipped with a PTFE-coated magnetic stir bar was charged with 370 mg of the product from step A (0.90 mmol, 1 eq.), dissolved in 3.0 mL of dry THF, and then at room temperature. 990 μl of TBAF (1 M in THF, 0.99 mmol, 1.1 eq.) was added dropwise. The resulting mixture was stirred at room temperature. for 15 minutes, during which the reaction reached a state of complete transformation. The reaction mixture was quenched by adding 200 μl of conc. NH4C1 solution, and then celite was added to the reaction mixture and the volatiles were removed under reduced pressure. The crude product was then purified by flash column chromatography using DCM and MeOH (1.2% NH ₃ ) as eluents to obtain 150 mg (60%) of the title product.

Enantiomers were separated using chiral chromatography. Column: AD-H, eluents: heptane/EtOH + 0.1% DEA; the early eluting enantiomer was collected as 6g with 99.8% ee, and the later eluting enantiomer was collected as 6h with 99.4% ee.

Synthesis 6i: 4-[1-[(dimethylamino)methyl]-3-bicyclo[1.1.1]pentanyl]phenol.

Step A: 1 -(4-methoxyphenyl)-N,M-dimethylbicyclo[1.1.1]pentane-3-carboxamide.

300 mg of 1-(4-methoxyphenyl)bicyclo[1.1.1]pentane-3-carboxylic acid (1.38 mmol, 1 eq.) and 278 mg of NN-diethalethanamine (2.75 mmol, 2 eq.) were mixed in EtOAc (3 ml/ mmol), then 1312 mg of 2.4.6-tripropyl-1.3.5.2l ^L {5;.4l ^L {5;.6/. ^L {5;-'grioxatri(|)oc(|)inane 2,4,6-trioxide (50 wt.% in EtOAc, 2.06 mmol, 1.5 eq.), and the mixture was stirred at RT. within 40 min. After reaction time, 1.03 mL of N-methylmethanamine (2 M in MeOH, 2.06 mmol, 1.5 eq.) was added and the mixture was stirred at room temperature. until complete conversion was observed (30 min). The reaction mixture was diluted with DCM, washed with conc. NaHCO ₃ solution, then the organic phase was washed with conc. NaCl, dried over MgSO ₄ , filtered, concentrated, and dried in vacuo to give 346 mg (quantity) of the title product as a peach-colored solid.

1H NMR (500 MHz, DMSOHO δ ppm 7.14 (m, 2H), 6.86 (m, 2H), 3.72 (s, 3H), 3.08 (s, 3H), 2.81 (s, 3H), 2.26 (s , 6H); ¹³ C NMR (125 MHz, DMSOL) δ ppm 168.9, 158.6, 132.5, 127.6, 114.1, 55.5, 54.2, 42.0, 39.0, 37.4, 35.9; HRMS-ERI (m/z): [ M+H] ⁺ , calculated for C ₁₅ H ₂₀ NO ₂ : 246.1488; found: 246.1495.

Step B: 1-[3-(4-methoxyphenyl)-1-bicyclo[1.1.1]pentanyl]-N,N-dimethylmethanamine.

289 mg of the product from step A (1.18 mmol, 1 eq.) was dissolved in THF (5 ml/mmol), then 2.36 ml of LiAlH ₄ (1 M in THF, 2.36 mmol, 2 eq.) was added at ambient temperature under nitrogen atmosphere. , and the mixture was stirred until complete conversion was achieved (approx. 1 hour). The mixture was cooled to 0°C, then quenched with conc. NH4C1 solution. After quenching, ~5 mL water and ~10 mL EtOAc were added and the mixture was shaken well. 2 M HCl was added and the (acidic) aqueous phase was separated and the organic phase was then extracted with additional 2 M HC1. The combined aqueous phases were made basic with 2 M NaOH and extracted with DCM.

The combined organic phases were washed with brine, dried over MgSO ₄ , concentrated and dried in vacuo. 219 mg (80%) of the target product was obtained in the form of a viscous oil.

1H NMR (500 MHz, DMSOHO δ ppm 7.10 (m, 2H), 6.84 (m, 2H), 3.71 (s, 3H), 2.35 (s, 2H), 2.16 (s, 6H), 1.89 (s , 6H); ¹³ C NMR (125 MHz, DMSO^) δ ppm 158.4, 133.4, 127.4, 114, 60.8, 55.5, 52.9, 46.6, 41.9, 38.0; HRMS-ERI (m/z): [M +H]+, calculated for C ₁₅ H ₂₂ NO: 232.1696; found: 232.1700.

Step C: 4-[1-[(dimethylamino)methyl]-3-bicyclo[1.1.1]pentanyl]phenol.

mg of the product from step B (0.22 mmol, 1 eq.) was dissolved in DCM (5 ml/mmol), then 0.65 ml of BBr ₃ (1 M in DCM, 0.65 mmol, 3 eq.) was added under a nitrogen atmosphere at 0°C, and the mixture was stirred for 30 min at 0°C and at room temperature. until complete conversion is achieved (approx. 45 min). DCM was added, the mixture was poured into a NaHCO3 solution, stirred for several minutes and then brought to neutral with conc. NH4C1 solution. The organic phase was separated and washed with brine, dried over MgSO ₄ , concentrated and dried in vacuo. 47 mg (quantit.) of crude target product were obtained in the form of a viscous oil.

1H NMR (400 MHz, CDC13) δ ppm 7.07 (d, 2H), 6.81 (d, 2H), 5.18 (br. s, 1H), 3.83 (s, 2H), 3.17/3.16 (s+s, 6H), 2.23 (s, 6H); ¹³ C NMR (100 MHz, CDC13) δ ppm 155.0, 131.3, 127.3, 115.4, 59.6, 54.7, 53.4, 46.0, 44.1; LC-MS-ESI (m/z): [M+H] ⁺ , calculated for C14H ₂₀ NO: 218.2; found: 218.2.

Synthesis of 6j: ^NiDimethyl-3-(4-triisopropylsilyloxyphenyl)prop-2-yn-1-amine.

500 mg of 4-iodophenol (2.27 mmol, 1 eq.) and 628 mg of K ₂ CO ₃ (4.55 mmol, 2 eq.) were mixed in acetonitrile (5 ml/mmol), then at room temperature. 526 mg of chloro(triisopropyl)silane (2.73 mmol, 1.2 eq.) was added and the mixture was stirred for 1 hour. The reaction mixture was filtered through a pad of celite, then 236 mg of C.X-dimethylprop-2-yne-1-amine ( 2.84 mmol, 1.25 eq.), 50 mg Pd(PPh ₃ ) ₂ C1 ₂ (0.11 mmol, 0.05 eq.), 22 mg CuI (0.11 mmol, 0.05 eq.) and 2.27 ml of isopropylpropan-2-amine (1 ml /mmol), and then the mixture was stirred at 50°C for 3 hours. The reaction mixture was concentrated on celite and purified by flash column chromatography using heptane and EtOAc as eluents to obtain 449 mg (60%) of the title product as yellow oil.

1H NMR (500 MHz, DMSO-de) δ ppm 7.32 (dm, 2H), 6.84 (dm, 2H), 3.41 (s, 2H), 2.22 (s, 6H), 1.24 (m, 3H), 1.06 (d, 18H); ¹³ C NMR (125 MHz, DMSO-de) 156, 133.5, 120.3, 116, 85.1/84.5, 44.3, 18.3, 12.4; MSVRERI (m/z): [M+H] ⁺ , calculated for C ₂₀ H ₃₄ NOSi: 332.2404; found: 332.2405.

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Synthesis of 6k: 2-Fluoro-4-{3-[(4-methoxyphenyl)methoxy]prop-1-yn-1-yl}phenol.

Stage A: 4-bromo-2-fluorophenylacetate.

To a solution of 4-bromo-2-fluorophenol (2.29 ml, 20.9 mmol, 1 equiv.) in dichloromethane (60 ml) was added 4(dimethylamino)pyridine (5.12 g, 41.9 mmol, 2 equiv.) and acetic anhydride (4.94 ml , 52.4 mmol, 2.5 eq.), and the mixture was stirred at ambient temperature for 18 hours. The reaction mixture was concentrated, then partitioned between dichloromethane and water, washed with brine, separated (phase separator) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 80 g RediSep™ silica gel cartridge) eluting with a gradient of 0-10% ethyl acetate in isoheptane gave the title product as a clear oil (5.1 g, 21.9 mmol, >100%) .

1H NMR (400 MHz, DMSOL) δ 7.75 (dd, J=10.0, 2.3 Hz, 1H), 7.48 (ddd, J=8.6, 2.3, 1.3 Hz, 1H), 7.30 (t, J=8.5 Hz, 1H) , 2.41 (s, 3H).

Step B: 1-methoxy-4-[(prop-2-yn-1-yloxy)methyl]benzene.

To a stirred suspension of sodium hydride (60% dispersion; 856 mg, 21.4 mmol, 1.2 eq.) in tetrahydrofuran (25 ml), cooled to 0°C, was added a solution of propargyl alcohol (1.04 ml, 17.8 mmol, 1 eq.) in tetrahydrofuran (10 ml) followed by the addition of 1-(bromomethyl)-4-methoxybenzene (3.09 ml, 21.4 mmol, 1.2 eq.), and the mixture was stirred at ambient temperature for 18 hours. The reaction mixture was partitioned between ethyl acetate and saturated aqueous chloride solution ammonium, and the organic phase was dried (magnesium sulfate), filtered and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 80 g RediSep™ silica gel cartridge) eluting with a gradient of 0-10% ethyl acetate in isoheptane gave the title product as a clear oil (2.44 g, 13.9 mmol, 78%).

1H NMR (400 MHz, DMSOO δ 7.31-7.18 (m, 2H), 6.97-6.86 (m, 2H), 4.44 (s, 2H), 4.13 (d, J=2.4 Hz, 2H), 3.75 (s, 3H ), 3.48 (t, J=2.4 Hz, 1H).

Step C: 2-fluoro-4-{3-[(4-methoxyphenyl)methoxy]prop-1-yn-1-yl}phenylacetate.

The product from stage A (1.68 g, 7.23 mmol, 1 eq.), triethylamine (18 ml), the product from stage B (1.91 g, 10.8 mmol, 1.5 eq.), copper iodide ( [) (275 mg, 1.45 mmol, 0.2 eq.) and tetrakis(triphenylphosphine)palladium(0) (835 mg, 0.72 mmol, 0.1 eq.), and the mixture was bubbled with nitrogen (10 min), after which the tube was sealed and heated at 90°C for 4 hours. The reaction mixture was allowed to cool to ambient temperature, then partitioned between dichloromethane and brine, and the organic phase was separated, dried (PTFE phase separator) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 40 g RediSep™ silica gel cartridge) eluting with a gradient of 0-30% ethyl acetate in isoheptane gave the title product as an orange solid (2.11 g, 6.43 mmol, 89%).

1H NMR (400 MHz, DMSOO δ 7.55 (dd, J=11.1, 1.7 Hz, 1H), 7.44-7.28 (m, 4H), 6.96 - 6.91 (m, 2H), 4.53 (s, 2H), 4.39 (s , 2H), 3.76 (s, 3H), 2.34 (s, 3H).

Step D: 2-fluoro-4-{3-[(4-methoxyphenyl)methoxy]prop-1-yn-1-yl}phenol.

To a solution of the product from step C (200 mg, 0.61 mmol, 1 eq) in methanol (15 ml) was added potassium carbonate (253 mg, 1.83 mmol, 3 eq) and the mixture was stirred at ambient temperature for 3 h. The reaction mixture was concentrated in vacuo, partitioned between dichloromethane and dilute aqueous hydrochloric acid, and the organic phase was separated (PTFE phase separator) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 12 g RediSep™ silica gel cartridge) eluting with a gradient of 0-40% ethyl acetate in isoheptane gave the title product as a yellow gum (128 mg, 0.45 mmol, 73%).

LC/MS (C17H15FO3) 285 [M-H] ^- ; RT 2.09 (LCMS-VC).

1H NMR (400 MHz, DMSO-de) δ 10.37 (s, 1H), 7.32-7.24 (m, 3H), 7.16-7.10 (m, 1H), 6.98-6.90 (m, 3H), 4.50 (s, 2H ), 4.34 (s, 2H), 3.75 (s, 3H).

Synthesis of 7c: 3,6-Dichloro-4-(propan-2-yl)pyridazine.

3,6-Dichloropyridazine (5 g, 33.6 mmol, 1 eq.), silver nitrate (5.7 g, 33.6 mmol, 1 eq.) and isobutyric acid (2.96 g, 33.6 mmol, 1 eq.) were suspended in water (100 ml ). The mixture was heated to 50°C and sulfuric acid (9.88 g, 101 mmol, 3 eq.) was added. The mixture was heated to 60°C and 1.6 M aqueous ammonium persulfate (62.9 mL, 101 mmol, 3 eq.) was added dropwise and the reaction mixture was heated at 70°C for 30 min. The mixture was allowed to cool to ambient temperature, then cooled to 0°C and slowly neutralized with concentrated ammonium hydroxide to pH 7, and then adjusted to pH 8-9. A yellow precipitate formed which was filtered and washed with water and ethyl acetate to give a white solid. The filtrates were combined, the layers were separated, and the aqueous phase was extracted with ethyl acetate (100 ml). The combined organics were washed with brine (100 ml), dried (magnesium sulfate) and concentrated in vacuo to give a yellow oil. Purification using automated flash column chromatography (CombiFlash Rf, 80 g silica gel

- 70 046336 RediSep™ cartridge) eluting with a gradient of 0-5% ethyl acetate in heptane gave the title product as a colorless oil (4.61 g, 24.1 mmol, 72%).

LC/MS (C ₇ H ₈ Cl ₂ N ₂ ) 191 [M+H]+; RT 1.11 (LCMS-V-BI).

1H NMR (400 MHz, DMSOL) δ 8.00 (d, J=0.6 Hz, 1H), 3.16 (h, 1H), 1.26 (d, J=6.8 Hz, 6H).

Synthesis 7e: 3,6-Dichloro-4-methyl-5-(propan-2-yl)pyridazine.

A suspension of 3,6-dichloro-4-methylpyridazine (0.5 g, 3.07 mmol, 1 eq.), isobutyric acid (0.28 ml, 3.07 mmol, 1 eq.) and silver nitrate (0.52 g, 3.07 mmol, 1 eq.) in water (10 ml) was heated to 50°C. Sulfuric acid (0.49 mL, 9.2 mmol, 3 eq.) was added and the mixture was heated to 60°C. 1.6 M aqueous ammonium persulfate (5.75 mL, 9.2 mmol, 3 eq) was added dropwise and the mixture was stirred for 30 min and then allowed to cool to ambient temperature. Performing ammonium hydroxide neutralization, celite filtration, and ethyl acetate elution resulted in a biphasic mixture. The layers were separated, the aqueous phase was extracted with ethyl acetate, and the combined organics were dried (magnesium sulfate) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 40 g RediSep™ silica gel cartridge) eluting with a gradient of 0-15% ethyl acetate in isoheptane provided the title product as a white solid (476 mg, 2.32 mmol, 76%) . LC/MS (C ₈ Hi ₀ C1 ₂ N ₂ ) 205 [M+H]+; RT 1.15 (LCMS-V-BI).

1H NMR (400 MHz, DMSOL ₆ ) δ 3.57 (hept, J=7.2 Hz, 1H), 2.49 (s, 3H), 1.35 (d, J=7.2 Hz, 6H).

Synthesis of 7f: 3,6-Dichloro-4-ethyl-5-methylpyridazine.

Silver nitrate (5.21 g, 30.7 mmol, 1 eq.), and the mixture was heated to 50°C. Sulfuric acid (4.91 mL, 92 mmol, 3 eq.) was added dropwise and the reaction mixture was heated to 60°C. Then, 1.6 M aqueous ammonium persulfate (57.5 mL, 92 mmol, 3 eq.) was added dropwise over 20 min. The reaction mixture was heated to 70°C for 30 min and then allowed to cool to ambient temperature. The mixture was made alkaline to pH 8 using conc. ammonium hydroxide solution, and the product was extracted with ethyl acetate (100 ml x 2). The combined organic extracts were dried (magnesium sulfate) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 120 g RediSep™ silica gel cartridge) eluting with a gradient of 0-25% ethyl acetate in isoheptane provided the title product as a white solid (4.11 g, 21.5 mmol, 70%) .

LC/MS (C ₇ H ₈ Cl ₂ N ₂ ) 191 [M+H]+; RT 2.17 (LCMS-VC).

1H NMR (400 MHz, DMSOL ₆ ) δ 2.82 (q, J=7.6 Hz, 2H), 2.44 (s, 3H), 1.13 (t, J=7.6 Hz, 3H).

Synthesis of 7g: 4-[(Benzyloxy)methyl]-3,6-dichloro-5-methylpyridazine.

Silver nitrate (5.21 g, 30.7 mmol, 1 eq.), and the mixture was heated to 50°C. Sulfuric acid (4.91 mL, 92 mmol, 3 eq.) was added and the mixture was heated to 60°C. 1.6 M aqueous ammonium persulfate (57.5 mL, 92 mmol, 3 eq.) was added dropwise and the reaction mixture was heated at 70°C for 30 min. The mixture was allowed to cool to ambient temperature and made alkaline to pH 8 conc. an aqueous solution of ammonium hydroxide. The mixture was extracted with ethyl acetate (2 x 150 ml) and the combined organics were washed with brine (200 ml), dried (magnesium sulfate) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 120 g RediSep™ silica gel cartridge) eluting with a gradient of 0-25% ethyl acetate in isoheptane gave the title product as a yellow oil (4.67 g, 16.5 mmol, 54%).

LC/MS (C ₁₃ H ₁₂ Cl ₂ N ₂ O) 283 [M+H]+; RT 1.26 (LCMS-V-BI).

1H NMR (400 MHz, DMSOZH) δ 7.42-7.27 (m, 5H), 4.71 (s, 2H), 4.60 (s, 2H), 2.46 (s, 3H).

Synthesis 7h: 3,6-Dichloro-4-cyclopropyl-5-methylpyridazine.

Silver nitrate (5.09 g, 29.9 mmol, 1 eq.), and the mixture was heated to 50°C. Sulfuric acid (4.79 mL, 89.8 mmol, 3 eq.) was added to the mixture and the mixture was heated to 60°C. 1.6 M aqueous ammonium persulfate (56.1 mL, 89.8 mmol, 3 eq) was added dropwise over 20 min, and the mixture was then heated to 70°C and stirred for 30 min. The mixture was allowed to cool to ambient temperature and made alkaline to pH 8 conc. ammonium hydroxide solution. The product was extracted into the organic phase using ethyl acetate (2 x 200 ml) and the combined organic extracts were washed with brine, dried (magnesium sulfate) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 120 g RediSep™ silica gel cartridge) eluting with a gradient of 0-30% ethyl acetate in isoheptane provided the title product as a white solid (3.99 g, 19.7 mmol, 66%) .

LC/MS (C ₈ H ₈ Cl ₂ N ₂ ) 203 [M+H]+; RT 1.09 (LCMS-V-BI).

1H NMR (400 MHz, DMSOZH) δ 2.49 (d, J=0.9 Hz, 3H), 1.98-1.88 (m, 1H), 1.26-1.11 (m, 2H), 0.78-0.67 (m, 2H).

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Synthesis 71: 3,6-dichloro-4-cyclopropylpyridazine.

To a suspension of 3,6-dichloropyridazine (2 g, 13.4 mmol, 1 eq.) and cyclopropanecarboxylic acid (1.07 ml, 13.4 mmol, 1 eq.) in water (85 ml) was added silver nitrate (2.28 g, 13.4 mmol, 1 eq. .), and the mixture was heated to 50°C. Sulfuric acid (2.15 mL, 40.3 mmol, 3 eq.) was added and the mixture was heated to 60°C. 1.6 M aqueous ammonium persulfate (25.2 mL, 40.3 mmol, 3 eq) was added dropwise over 15 min and the reaction mixture was then heated at 70°C for 30 min. The mixture was allowed to cool to ambient temperature and made alkaline to pH 10 conc. ammonium hydroxide solution. The product was extracted into ethyl acetate (2x150 ml) and the extracts were washed with brine, dried (magnesium sulfate) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 80 g RediSep™ silica gel cartridge) eluting with a gradient of 0-20% ethyl acetate in isoheptane gave the title product as a white solid (985 mg, 5.21 mmol, 39%). LC/MS (C ₇ H ₆ Cl ₂ N ₂ ) 189 [M+H]+; RT 1.00 (LCMS-V-BI).

1H NMR (400 MHz, DMSO<) δ 7.61 (d, J=0.6 Hz, 1H), 2.19-2.09 (m, 1H), 1.25-1.19 (m, 2H), 1.08-1.03 (m, 2H).

Synthesis of 7m: 4-[3-(benzyloxy)propyl]-3,6-dichloro-5-methylpyridazine.

Step A: [(pent-4-yn-1-yloxy)methyl]benzene.

4-pentin-1-ol (5.53 mL, 59.4 mmol, 1 eq.) and tetrahydrofuran (50 mL) were charged into an oven-dried flask, and the solution was cooled to 0°C. Sodium hydride (60%; 2.85 g, 119 mmol, 2 eq.) was added in portions and the reaction mixture was allowed to stir for 30 min at 0°C. Benzyl bromide (7.42 mL, 62.4 mmol, 1.05 eq) was added dropwise and the reaction mixture was allowed to warm to ambient temperature and stirred for 16 h. The reaction mixture was cooled to 0°C and quenched by adding saturated aqueous ammonium chloride (20 ml). Water (20 ml) was added and the layers were separated. The aqueous layer was extracted with ethyl acetate (2x50 ml) and the combined organics were washed with brine (50 ml), dried (magnesium sulfate) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 120 g RediSep™ silica gel cartridge) eluting with a gradient of 0-10% ethyl acetate in isoheptane gave the title product as a yellow oil (8.93 g, 51.3 mmol, 86%).

LC/MS (C ₁₂ H ₁₄ O) 175 [M+H]+; RT 1.25 (LCMS-V-BI).

1H NMR (400 MHz, CDCl ₃ ) δ 7.42-7.21 (m, 5H), 4.52 (s, 2H), 3.57 (t, J=6.2 Hz, 2H), 2.32 (td, J=7.1, 2.7 Hz, 2H ), 1.94 (t, J=2.7 Hz, 1H), 1.84 (tt, J=7.1, 6.2 Hz, 2H).

Step B: [(hex-4-yn-1-yloxy)methyl]benzene.

A solution of the product from step A (3.9 g, 22.4 mmol, 1 eq.) in tetrahydrofuran (30 ml) was loaded into an oven-dried flask under a nitrogen atmosphere. The reaction mixture was cooled to -78°C and n-butyllithium (2.2 M in hexane; 12.2 mL, 26.9 mmol, 1.2 eq.) was added over 30 min and the mixture was stirred at -78°C for 1 h. Dropwise iodomethane (2.09 mL, 33.6 mmol, 1.5 eq.) was added and the resulting mixture was allowed to warm to 0°C over 2 hours. The reaction mixture was quenched by adding saturated aqueous ammonium chloride (20 mL), then diluted with water (20 ml) and extracted with ethyl acetate (3x20 ml). The combined organic extracts were washed successively with 2M aqueous sodium thiosulfate (50 ml) and brine (50 ml), dried (magnesium sulfate) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 80 g RediSep™ silica gel cartridge) eluting with a gradient of 0-10% ethyl acetate in isoheptane gave the title product as a straw-yellow liquid (3.61 g, 19.2 mmol, 86%). LC/MS (C ₁₃ H ₁₆ O) without ionization; RT 1.15 (LCMS-V-BI).

1H NMR (400 MHz, DMSO<) δ 7.43-7.19 (m, 5H), 4.46 (s, 2H), 3.48 (t, J=6.3 Hz, 2H), 2.23-2.14 (m, 2H), 1.72 (t , 3H), 1.71-1.62 (m, 2H).

Step C: 4-[3-(benzyloxy)propyl]-3,6-dichloro-5-methylpyridazine.

A solution of dichloro-1,2,4,5-tetrazine (2.5 g, 16.6 mmol, 1 eq.) and the product from step B (3.74 g, 19.9 mmol, 1.2 eq.) in toluene (15 ml) was heated in a sealed tube at 160°C for 20 hours. The mixture was allowed to cool to ambient temperature and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 80 g RediSep™ silica gel cartridge) eluting with a gradient of 0-35% ethyl acetate in isoheptane gave the title product as an orange oil (3.06 g, 9.83 mmol, 62%).

LC/MS (C ₁₅ H ₁₆ Cl ₂ N ₂ O) 311 [M+H]+; RT 1.35 (LCMS-V-BI).

1H NMR (400 MHz, DMSO<) δ 7.47-7.14 (m, 5H), 4.48 (s, 2H), 3.53 (t, J=5.9 Hz, 2H), 2.96-2.81 (m, 2H), 2.42 (s , 3H), 1.87-1.72 (m, 2H).

Synthesis of 8ca and 8cb: 6-chloro-5-(propan-2-yl)pyridazin-3-amine and 6-chloro-4-(propan-2-yl)pyridazin-3amine.

Synthesis product 7c (3.61 g, 18.9 mmol, 1 equiv.) was suspended in ammonia water (20 ml) and heated at 150°C for 4 h under microwave irradiation. The solids were collected by filtration, washed with water and dried to give 6-chloro-5-(propan-2-yl)pyridazin-3-amine (2.06 g, 12 mmol, 64%) as an off-white solid [~9 :1 mixture with another regioisomer] (product

- 72 046336 synthesis of 8ca).

LC/MS (C7H10C1N3) 172 [M+H]+; RT 0.81 (LCMS-V-BI).

1H NMR (400 MHz, DMSO-dg) δ 6.76 (d, J=0.7 Hz, 1H), 6.51 (s, 2H), 3.06-2.82 (m, 1H), 1.18 (d, 6H).

The filtrate was concentrated in vacuo and the crude solid was extracted with dichloromethane, filtered and evaporated to give 6-chloro-4-isopropylpyridazin-3-amine (447 mg, 2.6 mmol, 14%) as a yellow oil [~7:3 mixture with other regioisomer ] (synthesis product 8cb). LC/MS (C ₇ H ₁₀ ClN ₃ ) 172 [M+H]+; RT 0.79 (LCMS-V-BI).

1H NMR (400 MHz, DMSO^) δ 7.23 (s, 1H), 6.53 (s, 2H), 2.95-2.80 (m, 1H), 1.15 (d, 6H).

Synthesis of 8d: 6-chloro-5-methylpyridazin-3-amine and 6-chloro-4-methylpyridazin-3-amine.

A suspension of 3,6-dichloro-4-methylpyridazine (2 g, 12.3 mmol, 1 eq.) in ammonia water (40 ml) was heated in a high pressure reactor at 150°C for 6 h, and then allowed to cool to ambient temperature . The resulting precipitate was filtered, washed with water (10 ml) and dried to obtain the target product in the form of thin brown needles (1.31 g, 0.01 mol, 74%) [regioisomer ratio ~ 6:4]. The mixture of isomers was used in the next step without further purification. LC/MS (C ₅ H ₆ ClN ₃ ) without ionization; RT 0.28 (LCMS-V-BI).

Synthesis 8e: 6-chloro-4-methyl-5-(propan-2-yl)pyridazin-3-amine and 6-chloro-5-methyl-4-(propan-2yl)pyridazin-3-amine.

Synthesis product 7e (480 mg, 2.34 mmol, 1 eq.) was introduced into the high-pressure reaction vessel. Ammonia water (15 ml) was added, the vessel was sealed and the mixture was stirred at 160°C for 6 hours. The reaction mixture was then allowed to cool to ambient temperature, the vessel was opened and the reaction mixture was concentrated in vacuo, and the mixture of isomers was used in the next step without cleaning.

LC/MS (C ₈ H ₁₂ ClN ₃ ) 186 [M+H]+; RT 0.88 (LCMS-V-BI).

Synthesis of 8f: 6-chloro-5-ethyl-4-methylpyridazin-3-amine and 6-chloro-4-ethyl-5-methylpyridazin-3-amine.

A suspension of synthesis product 7f (4 g, 21 mmol, 1 eq) in ammonia water (15 ml) was stirred at 160°C in a stoppered flask for 24 hours. The mixture was allowed to cool to ambient temperature and a brown solid was observed to precipitate. The solid was collected by filtration, washed with water and dried in vacuo to give the desired products as a brown solid (1.4 g, 8.16 mmol, 39%) [2:1 mixture of regioisomers]. The mixture of isomers was used in the next step without purification.

LC/MS (C ₇ H ₁₀ ClN ₃ ) 172 [M+H]+; RT 1.53 (LCMS-VC).

Synthesis of 8g: 5-[(benzyloxy)methyl]-6-chloro-4-methylpyridazin-3-amine and 4-[(benzyloxy)methyl]-6chloro-5-methylpyridazin-3-amine.

Synthesis product 7g (600 mg, 2.12 mmol, 1 equiv.) was suspended as an oil in ammonia water (7 ml) and heated at 160°C for 5 h in a sealed tube. The mixture was allowed to cool to ambient temperature and a brown solid was observed to precipitate. The solid was collected by filtration, washed with water and dried in vacuo to give the title product (455 mg, 1.73 mmol, 81%) as a mixture of two regioisomers, which was directly used in the next step. LC/MS (C ₁₃ H ₁₄ ClN ₃ O) 264 [M+H]+; RT 1.02 (LCMS^-B1).

Synthesis of 8ha and 8hb: 6-chloro-5-cyclopropyl-4-methylpyridazin-3-amine and 6-chloro-4-cyclopropyl-5methylpyridazin-3-amine.

Synthesis 7h (3.99 g, 19.7 mmol, 1 eq.) was suspended in ammonia water (10 mL) and heated in a sealed tube for 20 h. The mixture was allowed to cool to ambient temperature and the precipitated solid was collected by filtration and dried to give 6-chloro-5cyclopropyl-4-methylpyridazin-3-amine (2.44 g, 13.3 mmol, 68%) [synthesis product 8ha], which was directly used in the next step without further purification.

LC/MS (C ₈ H ₁₀ ClN ₃ ) 184 [M+H]+; RT 0.80 (LCMS-^B1).

The filtrate was concentrated under reduced pressure and then evaporated with toluene (x3) to remove residual water to obtain a crude material (2.00 g, 10.89 mmol, 55%) containing 6-chloro-4cyclopropyl-5-methylpyridazin-3-amine, which was used directly in the following stages without additional purification [synthesis product 8hb].

LC/MS (C ₈ H ₁₀ ClN ₃ ) 184 [M+H]+; RT 0.79 (LCMS-^B1).

Synthesis of 8ia and 8ib: (6-amino-3-chloro-5-methylpyridazin-4-yl)methanol and (3-amino-6-chloro-5methylpyridazin-4-yl)methanol.

To a cooled solution of synthesis product 8g (3.1 g, 11.8 mmol, 1 eq.) in dichloromethane (100 ml), a solution of boron trichloride (58.8 ml, 1 M, 0.06 mol, 5 eq.) was added dropwise and the mixture was stirred at ambient temperature environment for 2 hours. The reaction mixture was cooled to 0-5°C and carefully quenched by adding methanol (30 ml). Additional methanol (30 ml) was added and the mixture was refluxed for 30 minutes. The solvent was removed in vacuo and the crude product was purified using automated flash column chromatography (CombiFlash Rf, 120 g RediSep™ silica gel cartridge) eluting with a gradient of 0-17.5% methanol in dichloromethane with

- 73 046336 obtaining target regioisomeric products:

(3-Amino-6-chloro-5-methylpyridazin-4-yl)methanol as a beige solid (0.87 g, 5.0 mmol, 42%) [synthesis product 8ib].

LC/MS (C ₆ H ₈ ClN ₃ O) 174 [M+H]+; RT 0.26 (LCMS-V-BI).

1H NMR (400 MHz, DMSO^) δ 8.13 (s, 2H), 4.57 (s, 2H), 2.40 (s, 3H).

(6-Amino-3-chloro-5-methylpyridazin-4-yl)methanol as a beige solid (1.38 g, 7.9 mmol, 67%) [synthesis product 8ia].

LC/MS (C ₆ H8ClN ₃ O) 174 [M+H]+; RT 0.18 (LCMS-V-BI).

1H NMR (400 MHz, DMSO^) δ 8.18 (s, 2H), 4.56 (s, 2H), 2.31 (s, 3H).

Synthesis of 8ja: 6-chloro-5-(methoxymethyl)-4-methylpyridazin-3-amine.

Step A: 5-(bromomethyl)-6-chloro-4-methylpyridazin-3-amine.

To a suspension of synthesis product 8ia (960 mg, 5.53 mmol, 1 eq.) in dichloromethane (50 ml) was added phosphorus tribromide (0.62 ml, 6.34 mmol, 1.2 eq.), and the mixture was stirred for 1 h at ambient temperature. The reaction mixture was quenched by adding methanol (20 ml) and concentrated in vacuo to give a solid (3.2 g). The substance was directly used in the next step without further purification.

LC/MS (C ₆ H ₇ BrClN ₃ ) 238 [M+H]+; RT 0.81 (LCMS-V-BI).

Step B: 6-chloro-5-(methoxymethyl)-4-methylpyridazin-3-amine A solution of the crude product from Step A (780 mg) in methanol (10 ml) was heated at 120°C for 1 hour under microwave irradiation. The mixture was concentrated in vacuo and purified by automated flash column chromatography (CombiFlash Rf, 12 g RediSep™ silica gel cartridge) eluting with a gradient of 0-10% methanol in dichloromethane to give the title product as a brown oil (323 mg, 0.90 mmol, 66% for both stages).

LC/MS (C ₇ H ₁₀ ClN ₃ O) 188 [M+H]+; RT 0.61 (LCMS-V-BI).

Synthesis of 8jb: 6-chloro-4-(methoxymethyl)-5-methylpyridazin-3-amine.

Step A: 4-(bromomethyl)-6-chloro-5-methylpyridazin-3-amine.

To a suspension of synthesis product 8ib (500 mg, 2.88 mmol, 1 eq.) in dichloromethane (50 ml) was added phosphorus tribromide (0.32 ml, 3.46 mmol, 1.2 eq.), and the mixture was stirred for 1 h at ambient temperature.

The reaction mixture was quenched by adding methanol (20 ml) and then the volatile organics were removed in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 24 g RediSep™ silica gel cartridge) eluting with a gradient of 0-5% methanol in dichloromethane provided the title product as an orange solid (695 mg, 2.94 mmol, 100%) .

LC/MS (C ₆ H ₇ BrClN ₃ ) 238 [M+H]+; RT 0.82 (LCMS-V-BI).

1H NMR (400 MHz, DMSOL ₆ ) δ 7.84 (br s, including water peak), 4.76 (s, 2H), 2.38 (s, 3H).

Step B: 6-chloro-4-(methoxymethyl)-5-methylpyridazin-3-amine.

A solution of the product from step A (430 mg, 1.82 mmol, 1 eq.) in methanol (10 ml) was heated at 120°C for 30 min under microwave irradiation. The mixture was concentrated in vacuo and purified by automated flash column chromatography (CombiFlash Rf, 24 g RediSep™ silica gel cartridge) eluting with a gradient of 0-5% methanol in dichloromethane to give the title product as a purple crystalline solid (91 mg, 0.49 mmol, 27%).

LC/MS (C ₇ H ₁₀ ClN ₃ O) 188 [M+H]+; RT 0.70 (LCMS-V-BI).

1H NMR (400 MHz, DMSO^) δ 7.66 (s, 2H), 4.50 (s, 2H), 3.32 (s, 3H), 2.38 (s, 3H).

Synthesis of 8ka: 6-chloro-5-(ethoxymethyl)-4-methylpyridazin-3-amine.

A solution of synthesis product 8ja, step A (500 mg, 2.11 mmol, 1 equiv.) in ethanol (15 ml) was heated at 120°C for 1 h under microwave irradiation. The mixture was concentrated in vacuo and purified by automated flash column chromatography (CombiFlash Rf, 24 g RediSep™ silica gel cartridge) eluting with a gradient of 0-10% methanol in dichloromethane to give the title product as a viscous oil that crystallized when scratched (136 mg, 0.67 mmol, 32%).

LC/MS (C ₈ H ₁₂ ClN ₃ O) 202 [M+H]+; RT 0.63 (LCMS-V-BI).

1H NMR (400 MHz, DMSO^) δ 4.56 (s, 2H), 3.55 (q, J=7.0 Hz, 2H), 2.28 (s, 3H), 1.14 (t, J=7.0 Hz, 3H).

Synthesis of 8ma and 8mb: 5-[3-(benzyloxy)propyl]-6-chloro-4-methylpyridazin-3-amine and 4-[3(benzyloxy)propyl]-6-chloro-5-methylpyridazin-3-amine.

Synthesis product 7m (3.06 g, 9.83 mmol, 1 equiv.) and ammonia water (15 ml) were heated in a stoppered flask for 48 h at 160°C. The mixture was allowed to cool to ambient temperature and extracted with dichloromethane (2x100 ml). The organics were washed with brine (50 ml) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 80 g RediSep™ silica gel cartridge) eluting with a gradient of 0-100% ethyl acetate in isoheptane resulted in the desired regioisomeric products:

5-[3-(Benzyloxy)propyl]-6-chloro-4-methylpyridazin-3-amine as a beige solid

- 74 046336 (1.3 g, 4.46 mmol, 45%) [synthesis product 8ta].

LC/MS (C ₁₅ H ₁₈ ClN ₃ O) 292 [M+H]+; RT 1.12 (ZHMS-U-B1).

1H NMR (400 MHz, DMSOO δ 7.47-7.20 (m, 5H), 6.33 (s, 2H), 4.48 (s, 2H), 3.49 (t, J=6.0 Hz, 2H), 2.79-2.66 (m, 2H ), 2.08 (s, 3H), 1.84-1.65 (m, 2H).

4-[3-(Benzyloxy)propyl]-6-chloro-5-methylpyridazin-3-amine as a beige solid (500 mg, 1.71 mmol, 17%) [synthesis product 8mb].

LC/MS (C ₁₅ H ₁₈ ClN ₃ O) 292 [M+H]+; RT 1.15 (ZHMS-U-B1).

1H NMR (400 MHz, DMSOO δ 7.49-7.15 (m, 5H), 6.34 (s, 2H), 4.48 (s, 2H), 3.47 (t, J=6.2 Hz, 2H), 2.72-2.54 (m, 2H ), 2.22 (s, 3H), 1.83-1.61 (m, 2H).

Synthesis 9a: Na(6-chloro-4,5-dimethylpyridazin-3-yl)-1,3-benzothiazol-2-amine.

To a solution of 3-amino-6-chloro-4,5-dimethylpyridazine (450 mg, 2.86 mmol, 1 equiv.) and 2-bromo-1,3benzothiazole (672 mg, 3.14 mmol, 1.1 equiv.) in 1,4- dioxane (20 ml), sodium hydride (60% dispersion; 457 mg, 11.42 mmol, 4 eq.) was added in portions and the mixture was refluxed for 16 hours. The reaction mixture was allowed to cool to ambient temperature and then quenched with acetic acid. The mixture was diluted with ethyl acetate (200 ml) and washed with aqueous saturated sodium bicarbonate (100 ml) and brine (100 ml). The organic extract was dried (magnesium sulfate) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 40 g RediSep™ silica gel cartridge) eluting with a gradient of 0-3% methanol in dichloromethane provided the title product as a yellow solid (673 mg, 2.32 mmol, 81%) .

LC/MS (C13H11C1N4S) 291 [M+H]+; RT 1.22 (ZHMS-U-B1).

1H NMR (400 MHz, DMSO^) δ 11.80 (br s, 1H), 7.86 (d, J=7.75 Hz, 1H), 7.50 (s, 1H), 7.40 (td, J=1.28, 7.69 Hz, 1H) , 7.22 (t, J=7.67 Hz, 1H), 2.37 (s, 3H), 2.33 (s, 3H).

Synthesis 9b: Na(6-chloropyridazin-3-yl)-1,3-benzothiazol-2-amine.

To a solution of 3-amino-6-chloropyridazine (2 g, 15.4 mmol, 1 eq.) and 2-bromo-1,3-benzothiazole (3.64 g, 17.0 mmol, 1.1 eq.) in 1,4-dioxane (120 ml ) sodium hydride (60% dispersion; 2.47 g, 61.8 mmol, 4 eq.) was added in portions and the mixture was refluxed for 30 min. The reaction mixture was allowed to cool to ambient temperature and then cooled in an ice water bath and quenched with methanol. Removal of methanol by rotary evaporation caused precipitation of a solid. The mixture was cooled in ice water and the solid was collected by filtration, washed with ice-cold 1,4-dioxane (100 ml) and then diethyl ether (100 ml). Finally, the solid was suspended in water, the pH was adjusted to 7 and the solid was collected by filtration and dried in vacuo to give the expected product (1.94 g, 7.38 mmol, 48%).

LC/MS (C11H7C1N4S) 263 [M+H]+; RT 1.1 (ZHMS-U-B1).

1H NMR (400 MHz, DMSOO δ 12.03 (s, 1H), 8.00-7.93 (m, 1H), 7.82 (d, J=9.3 Hz, 1H), 7.69 (d, J=8.0 Hz, 1H), 7.59 ( d, J=9.3 Hz, 1H), 7.46-7.37 (m, 1H), 7.30-7.23 (m, 1H).

Synthesis of 9ca: Na[6-chloro-5-(propan-2-yl)pyridazin-3-yl]-1,3-benzothiazol-2-amine.

To a suspension of the synthesis product 8ca (2.06 g, 12 mmol, 1 eq.) in 1,4-dioxane (70 ml) was added 2bromo-1,3-benzothiazole (2.83 g, 13.2 mmol, 1.1 eq.) followed by the addition of hydride in portions sodium (60% dispersion; 0.96 g, 24.01 mmol, 2 eq.), and the mixture was heated at 90°C for 30 min. The reaction mixture was then allowed to cool to ambient temperature, after which the mixture was cooled in ice water, quenched by adding acetic acid and concentrated in vacuo. The resulting solid was triturated with water, filtered and dried in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 120 g RediSep™ silica gel cartridge) eluting with a gradient of 0-50% ethyl acetate in isoheptane gave a solid which was triturated with ethyl acetate, filtered and dried in vacuo to give the title product as an off-white solid (1.54 g, 5.05 mmol, 42%).

LC/MS (C ₁₄ H ₁₃ ClN ₄ S) 305 [M+H]+; RT 1.31 (ZHMS-U-B1).

1H NMR (400 MHz, DMSOO δ 11.87 (s, 1H), 7.99-7.91 (m, 1H), 7.69 (d, J=8.1 Hz, 1H), 7.51 (s, 1H), 7.42 (ddd, J=8.3 , 7.2, 1.3 Hz, 1H), 7.26 (ddd, J=8.2, 7.3, 1.2 Hz, 1H), 3.25-3.06 (m, 1H), 1.26 (d, J=6.8 Hz, 6H).

Synthesis of 9cb: N[6-chloro-4-(propan-2-yl)pyridazin-3-yl] - 1,3-benzothiazol-2-amine.

To a solution of synthesis product 8cb (47 mg, 2.6 mmol, 1 eq.) and 2-bromo-1,3-benzothiazole (669 mg, 3.13 mmol, 1.2 eq.) in 1,4-dioxane (10 ml) the hydride was added in portions sodium (60% dispersion; 208 mg, 5.21 mmol, 2 eq.), and the mixture was refluxed for 1 hour. The mixture was allowed to cool to ambient temperature, then cooled to 0°C and quenched by adding acetic acid. After evaporation, the residue was partitioned between ethyl acetate and water, and the organic phase was washed with brine, dried (magnesium sulfate) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 24 g RediSep™ silica gel cartridge) eluting with a gradient of 0-20% ethyl acetate in isoheptane gave the title product as a bright yellow solid (367 mg, 1.2 mmol, 46 %).

LC/MS (C ₁₄ H ₁₃ ClN ₄ S) 305 [M+H]+; RT 1.35 (ZHMS-U-B1).

- 75 046336 ¹ H NMR (400 MHz, DMSO-dg) δ 12.64 (s, 1H), 7.82 (br s, 1H), 7.61 (s, 1H), 7.40 (br s + t, J=7.3 Hz, 2H ), 7.22 (t, J=6.8 Hz, 1H), 3.49 (p, J=6.8 Hz, 1H), 1.25 (d, J=6.9 Hz, 6H).

Synthesis of 9da and 9db: N(6-chloro-5-methylpyridazin-3-yl)-1,3-benzothiazol-2-amine and N-(6-Χλορ-4methylpyridazin-3-yl)-1,3-benzothiazol- 2-am.

The hydride was added in portions to a suspension of synthesis product 8d (1.31 g, 9.1 mmol, 1 eq.) and 2-bromo-1,3-benzothiazole (2.14 g, 10.01 mmol, 1.1 eq.) in 1,4-dioxane (50 ml). sodium (60% dispersion; 0.73 g, 18.2 mmol, 2 eq.), and the mixture was refluxed for 1 hour. The reaction mixture was allowed to cool to ambient temperature, quenched with acetic acid, then neutralized with aqueous sodium bicarbonate. The mixture was extracted with ethyl acetate (100 ml x 3) and the combined organic extracts were washed with brine, dried (magnesium sulfate) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 120 g RediSep™ silica gel cartridge) eluting with a gradient of 0-80% ethyl acetate in isoheptane resulted in the desired products:

Na(6-chloro-4-methylpyridazin-3-yl)-1,3-benzothiazol-2-amine as a yellow solid (140 mg, 0.51 mmol, 5.6%) [synthesis product 9db].

LC/MS (C ₁₂ H ₉ ClN ₄ S) 277 [M+H] ⁺ ; RT 1.18 (LCMS-V-BI).

1H NMR (400 MHz, DMSOO δ 7.90 (s, 1H), 7.71 (s, 1H), 7.57 (br s, 1H), 7.41 (t, J=7.5 Hz, 1H), 7.24 (t, J=7.5 Hz , 1H), 2.39 (s, 3H).

Na(6-Chloro-5-methylpyridazin-3-yl)-1,3-benzothiazol-2-amine as a yellow solid (69 mg, 0.25 mmol, 2.7%) [synthesis product 9da].

LC/MS (C ₁₂ H ₉ ClN ₄ S) 277 [M+H] ⁺ ; RT 1.15 (LCMS-V-BI).

1H NMR (400 MHz, DMSOHO δ 11.92 (s, 1H), 7.96 (d, J=7.7 Hz, 1H), 7.69 (d, J=8.5 Hz, 1H), 7.48 (d, J=1.2 Hz, 1H) , 7.42 (ddd, J=8.2, 7.2, 1.3 Hz, 1H), 7.26 (ddd, J=8.2, 7.3, 1.2 Hz, 1H), 2.38 (d, J=1.0 Hz, 3H).

Synthesis of 9ea and 9eb: N[6-chloro-4-methyl-5-(propan-2-yl)pyridazin-3-yl]-1,3-benzothiazol-2-amine and N[6-Chloro-5-methyl -4-(propan-2-yl)pyridazin-3-yl] - 1,3-benzothiazol-2-amine.

The hydride was added in portions to a suspension of synthesis product 8e (434 mg, 2.34 mmol, 1 eq.) and 2-bromo-1,3-benzothiazole (551 mg, 2.57 mmol, 1.1 eq.) in 1,4-dioxane (10 ml). sodium (60% dispersion; 187 mg, 4.68 mmol, 2 eq.), and the mixture was refluxed for 1 hour. The reaction mixture was allowed to cool to ambient temperature and then quenched with acetic acid. The mixture was then diluted with aqueous sodium bicarbonate and the product was extracted with dichloromethane. The organics were washed with brine, dried (magnesium sulfate) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 40 g RediSep™ silica gel cartridge) eluting with a gradient of 0-30% ethyl acetate in isoheptane resulted in the desired regioisomeric products:

No.[6-chloro-5-methyl-4-(propan-2-yl)pyridazin-3-yl]-1,3-benzothiazol-2-amine as a yellow solid (114 mg, 0.36 mmol, 15%) [synthesis product 9eb].

LC/MS (C ₁₅ H ₁₅ ClN ₄ S) 319 [M+H] ⁺ ; RT 1.39 (LCMS-V-BI).

1H NMR (400 MHz, DMSOO δ 12.48 (s, 1H), 7.73 (d, J=7.8 Hz, 1H), 7.41-7.32 (m, 1H), 7.32-7.26 (m, 1H), 7.18 (t, J =7.5 Hz, 1H), 3.76-3.58 (m, 1H), 2.41 (s, 3H), 1.41 (d, J=7.0 Hz, 6H).

No.[6-Chloro-4-methyl-5-(propan-2-yl)pyridazin-3-yl]-1,3-benzothiazol-2-amine as an orange solid (300 mg, 0.94 mmol, 40%) [synthesis product 9ea].

LC/MS (C ₁₅ H ₁₅ ClN ₄ S) 319 [M+H] ⁺ ; RT 1.36 (LCMS-V-BI).

1H NMR (400 MHz, DMSOD ₆ ) δ 12.64 (br s, 1H), 7.91-7.78 (m, 1H), 7.46 (br s, 1H), 7.40 (t, 1H), 7.22 (t, 1H), 3.56 (h, 1H), 2.47 (s, 3H), 1.37 (d, J=7.2 Hz, 6H).

Synthesis of 9fa and 9fb: N(6-chloro-5-ethyl-4-methylpyridazin-3-yl)-1,3-benzothiazol-2-amine and N-(6-χλορ-4ethyl-5-methylpyridazin-3-yl )-1,3-benzothiazol-2-amine.

The hydride was added in portions to a suspension of synthesis product 8f (1.4 g, 8.16 mmol, 1 eq.) and 2-bromo-1,3-benzothiazole (2.1 g, 9.79 mmol, 1.2 eq.) in 1,4-dioxane (50 ml). sodium (60% dispersion; 1.31 g, 32.6 mmol, 4 eq.). The mixture was refluxed for 1.5 h, then allowed to cool to ambient temperature. The reaction mixture was quenched with acetic acid and neutralized with sodium bicarbonate. The mixture was extracted with ethyl acetate (x2) and the combined organic extracts were washed with brine, dried (magnesium sulfate) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 120 g RediSep™ silica gel cartridge) eluting with a gradient of 0-35% ethyl acetate in isoheptane resulted in the desired regioisomeric products:

Na(6-chloro-4-ethyl-5-methylpyridazin-3-yl)-1,3-benzothiazol-2-amine as a yellow solid (330 mg, 1.08 mmol, 13%) [synthesis product 9fb].

LC/MS (C ₁₄ H ₁₃ ClN ₄ S) 305 [M+H] ⁺ ; RT 1.31 (LCMS-V-BI).

1H NMR (400 MHz, DMSOD ₆ ) δ 7.82 (br s, 1H), 7.38 (br s + t, 2H), 7.21 (t, 1H), 2.91 (q, J=7.4 Hz, 2H), 2.38 (s , 3H), 1.12 (t, J=7.5 Hz, 3H).

No.(6-Chloro-5-ethyl-4-methylpyridazin-3-yl)-1,3-benzothiazol-2-amine as a yellow solid

- 76 046336 stva (750 mg, 2.46 mmol, 30%) [synthesis product 9fa].

LC/MS (C ₁₄ H ₁₃ ClN ₄ S) 305 [M+H]+; RT 1.30 (LCMS-V-BI).

1H NMR (400 MHz, DMSO^b) δ 7.86 (s, 1H), 7.49 (br s, 1H), 7.40 (t, 1H), 7.22 (t, 1H), 2.79 (q, J=7.5 Hz, 2H ), 2.41 (s, 3H), 1.14 (t, J=7.5 Hz, 3H).

Synthesis of 9ga and 9gb: H-{5-|(benzyloxy)methyl|-6-chloro-4-methylpyridazin-3-yl}-1.3-benzothiazol-2amine and N{4-[(benzyloxy)methyl]-6-chloro -5-methylpyridazin-3-yl}-1,3-benzothiazol-2-amine.

To a solution of synthesis product 8g (1.16 g, 4.4 mmol, 1 eq.) and 2-bromo-1,3-benzothiazole (1.22 g, 5.72 mmol, 1.3 eq.) in 1,4-dioxane (30 ml) the hydride was added in portions sodium (60% dispersion; 352 mg, 8.8 mmol, 2 eq.), and the reaction mixture was refluxed for 2 hours. The mixture was allowed to cool to ambient temperature, quenched with acetic acid and neutralized with sodium bicarbonate. The product was extracted using ethyl acetate (200 ml x 2) and washed with brine (100 ml). The organic extract was dried (magnesium sulfate) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 120 g RediSep™ silica gel cartridge) eluting with a gradient of 0-30% ethyl acetate in isoheptane resulted in the desired regioisomeric products:

NO{4-[(benzyloxy)methyl]-6-chloro-5-methylpyridazin-3-yl}-1,3-benzothiazol-2-amine as a yellow solid (250 mg, 0.63 mmol, 14%) [product synthesis 9gb].

LC/MS (C ₂₀ H ₁₇ ClN ₄ OS) 398 [M+H]+; RT 1.43 (LCMS-V-Bb

1H NMR (400 MHz, DMSOC6) δ 12.53 (br s, 1H), 7.84 (d, J=7.6 Hz, 1H), 7.55-7.27 (m, 7H), 7.23 (t, 1H), 4.89 (s, 2H ), 4.61 (s, 2H), 2.38 (s, 3H).

#{5-[(Benzyloxy)methyl]-6-chloro-4-methylpyridazin-3-yl}-1,3-benzothiazol-2-amine as a yellow solid (253 mg, 0.64 mmol, 14%) [product synthesis 9ga].

LC/MS (C ₂₀ H ₁₇ ClN ₄ OS) 398 [M+H]+; RT 1.39 (LCMS-V-BT).

1H NMR (400 MHz, DMSO^) δ 12.71 (br s, 1H), 7.85 (s, 1H), 7.45-7.27 (m, 7H), 7.26-7.15 (m, 1H), 4.67 (s, 2H), 4.60 (s, 2H), 2.44 (s, 3H).

Synthesis of 9ha: Na(6-chloro-5-cyclopropyl-4-methylpyridazin-3-yl)-1,3-benzothiazol-2-amine.

To a solution of the synthesis product 8ha (2.44 g, 13.3 mmol, 1 eq.) and 2-bromo-1,3-benzothiazole (3.7 g, 17.3 mmol, 1.3 eq.) in 1,4-dioxane (50 ml) the hydride was added in portions sodium (60% dispersion; 1.06 g, 26.6 mmol, 2 eq.), and the mixture was refluxed for 1 hour. The reaction mixture was allowed to cool to ambient temperature, then quenched with acetic acid and neutralized by adding sodium bicarbonate. The resulting precipitate was collected by filtration and dried in vacuo to give the target product (0.98 g, 3.09 mmol, 23%).

LC/MS (Ci ₅ Hi ₃ C1N ₄ S) 317 [M+H]+; RT 1.30 (LCMS-V-Bb

1H NMR (400 MHz, DMSOC6) δ 11.88 (br s, 1H), 7.82 (d, 1H), 7.47 (d, J=8.0 Hz, 1H), 7.39-7.32 (m, 1H), 7.21-7.13 (m , 1H), 2.47 (d, J=1.0 Hz, 3H), 1.93-1.79 (m, 1H), 1.23-1.08 (m, 2H), 0.69 (td, J=6.2, 4.4 Hz, 2H).

Synthesis of 9hb: H-(6-Chloro-4-cyclopropyl-5-methylpyridazin-3-yl)-1,3-benzothiazol-2-amine.

To a solution of 2-bromo-1,3-benzothiazole (2.8 g, 13.1 mmol, 1.2 eq.) and the synthesis product 8hb (2 g, 10.9 mmol, 1 eq.) in 1,4-dioxane (50 ml) the hydride was added in portions sodium (60% dispersion; 0.52 g, 21.8 mmol, 2 eq.), and the reaction mixture was refluxed for 1 hour. The mixture was allowed to cool to ambient temperature and concentrated in vacuo. The residue was taken up in ethyl acetate, filtered, and the solid was extracted with ethyl acetate. The combined organic extracts were washed with brine, dried (magnesium sulfate) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 40 g RediSep™ silica gel cartridge) eluting with a gradient of 0-40% ethyl acetate in isoheptane provided the title product as a yellow solid (219 mg, 0.69 mmol, 6%) .

LC/MS (Ci ₅ Hi ₃ C1N ₄ S) 317 [M+H]+; RT 1.28 (LCMS-^B1).

1H NMR (400 MHz, DMSOD ₆ ) δ 10.74 (br s, 1H), 8.05-7.85 (m, 1H), 7.78-7.55 (m, 1H), 7.42 (t, 1H), 7.25 (t, 1H), 2.47 (d, J=0.9 Hz, 4H), 2.01-1.89 (m, 1H), 1.14-1.02 (m, 2H), 0.72-0.60 (m, 2H).

Synthesis of 9ja: Na[6-chloro-5-(methoxymethyl)-4-methylpyridazin-3-yl]-1,3-benzothiazol-2-amine.

Sodium hydride (60 % dispersion; 138 mg, 3.44 mmol, 2 eq.), and the mixture was refluxed for 1.5 h. The mixture was cooled to ambient temperature, neutralized with acetic acid and concentrated in vacuo. The crude material was partitioned between dichloromethane and water, and the organic phase was dried (magnesium sulfate) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 40 g RediSep™ silica gel cartridge) eluting with a gradient of 0-50% ethyl acetate in isoheptane gave the title product as a bright yellow solid (85 mg, 0.26 mmol, 15 %).

LC/MS (C14H13C1N4OS) 321 [M+H]+; RT 1.19 (LCMS-^B1).

1H NMR (400 MHz, DMSOD ₆ ) δ 8.18-6.98 (m, 5H), 4.58 (s, 2H), 3.35 (s, 3H), 2.46 (s, 3H).

Synthesis of 9jb: N[6-chloro-4-(methoxymethyl)-5-methylpyridazin-3-yl] - 1,3-benzothiazol-2-amine.

- 77 046336

To a solution of synthesis product 8jb (140 mg, 0.81 mmol, 1 eq.) and 2-bromo-1,3-benzothiazole (224 mg, 1.05 mmol, 1.3 eq.) in 1,4-dioxane (7 ml) the hydride was added in portions sodium (60% dispersion; 58.1 mg, 2.42 mmol, 3 eq.), and the mixture was refluxed for 30 min. The reaction mixture was cooled to 0°C and slowly quenched with water, diluted with ethyl acetate and washed with sodium bicarbonate and brine. The organic phase was dried (magnesium sulfate) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 24 g RediSep™ silica gel cartridge) eluting with a gradient of 0-100% ethyl acetate in isoheptane provided the title product as a yellow solid (169 mg, 0.53 mmol, 58%) .

LC/MS (C ₁₄ H ₁₃ ClN ₄ OS) 321 [M+H]+; RT 1.23 (LCMS-V-BI).

¹ H NMR (400 MHz, DMSO-dg) δ 12.63 (s, 1H), 7.79 (s, 1H), 7.54-7.26 (m, 2H), 7.22 (s, 1H), 4.77 (s, 2H), 3.36 (s, 3H), 2.41 (s, 3H).

Synthesis of 9ka: Na[6-chloro-5-(ethoxymethyl)-4-methylpyridazin-3-yl]-1,3-benzothiazol-2-amine.

To a solution of the synthesis product 8ka (136 mg, 0.67 mmol, 1 eq.) and 2-bromo-1,3-benzothiazole (173 mg, 0.81 mmol, 1.2 eq.) in 1,4-dioxane (10 ml) the hydride was added in portions sodium (60% dispersion; 53.9 mg, 1.35 mmol, 2 eq.), and the mixture was refluxed for 1 hour. The reaction mixture was allowed to cool to ambient temperature and quenched with water. The mixture was partitioned between ethyl acetate and saturated aqueous sodium bicarbonate, and the organic phase was washed with brine, then dried (magnesium sulfate) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 24 g RediSep™ silica gel cartridge) eluting with a gradient of 0-40% ethyl acetate in isoheptane provided the title product as a yellow solid (113 mg, 0.34 mmol, 50%) .

LC/MS (C ₁₅ H ₁₅ ClN ₄ OS) 335 [M+H]+; RT 1.28 (LCMS-V-BI).

1H NMR (400 MHz, DMSOO δ 7.86 (s, 1H), 7.46 (br s, 1H), 7.41 (t, 1H), 7.23 (t, J=7.7 Hz, 1H), 4.61 (s, 2H), 3.56 (q, J=7.0 Hz, 2H), 2.47 (s, 3H), 1.16 (t, J=7.0 Hz, 3H).

Synthesis of 9ma: N{5-[3-(benzyloxy)propyl]-6-chloro-4-methylpyridazin-3-yl}-1,3-benzothiazol-2-amine.

To a solution of the synthesis product 8ta (1.3 g, 4.46 mmol, 1 equiv.) and 2-bromo-1,3-benzothiazole (1.14 g, 5.35 mmol, 1.2 equiv.) in 1,4-dioxane (50 ml) the hydride was added in portions sodium (60% dispersion; 356 mg, 8.91 mmol, 2 eq.), and the mixture was refluxed for 1 hour. The reaction mixture was allowed to cool to ambient temperature, then quenched by adding water and partitioned between ethyl acetate (100 ml) and sodium bicarbonate (50 ml). The organic phase was washed with brine (50 ml), dried (magnesium sulfate) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 80 g RediSep™ silica gel cartridge) eluting with a gradient of 0-55% ethyl acetate in isoheptane provided the title product as a cream solid (1.59 g, 3.74 mmol, 84%) .

LC/MS (C ₂₂ H ₂₁ ClN ₄ OS) 425 [M+H]+; RT 1.25 (LCMS-V-BI).

1H NMR (400 MHz, DMSOO δ 7.86 (s, 1H), 7.67-7.11 (m, 8H), 4.50 (s, 2H), 3.53 (t, J=6.0 Hz, 2H), 2.94-2.81 (m, 2H ), 2.41 (s, 3H), 1.89-1.72 (m, 2H).

Synthesis 10a: 1-(1-adamantylmethyl)-5-methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyrazole.

Step A: 1-(1-adamantylmethyl)-4-iodopyrazole.

A mixture of 35.9 g 1-adamantylmethanol (216 mmol), 73.48 g triphenylphosphine (280 mmol, 1.3 eq.), 54.25 g 4-iodo-1H-pyrazole (280 mmol, 1.3 eq.) and 64.4 g tert-butyl H-(tert-butoxycarbonyliminomethylene )carbamate (266 mmol. 1.3 eq.) in 1078 ml THF was stirred at room temperature. for 48 hours. After adding an additional 10.94 g of 4-iodo-1H-pyrazole (56 mmol, 0.26 eq.), 12.81 g of tert-butyl N(tert-butoxycarbonyliminomethylene)carbamate (53 mmol, 0.26 eq.) and 14.69 g of triphenylphosphine (56 mmol, 0.26 eq.), the reaction mixture was stirred at room temperature. for 24 hours, then concentrated, purified by flash column chromatography using DCM as eluant, and the resulting material was triturated in cold MeOH and filtered to obtain 53.6 g (73%) of the expected product.

Step B: 1-(1-adamantylmethyl)-4-iodo-5-methylpyrazole.

To 9.8 ml of diisopropylamine (69.5 mmol, 1.1 eq.) in 180 ml of THF at -78°C, 33.4 ml of 2.5 M butyllithium solution (84 mmol, 1.3 eq.) was added dropwise, and the mixture was stirred at -78°C for 0.5 h, treated with 22.0 g of the product from step A (64.28 mmol, 1 eq.) in 90 ml of THF, stirred at -78°C for 1 h, treated with 4.67 ml of methyl iodide (73.3 mmol, 1.14 eq.), and stirred at - 78°C for 18 hours. After quenching the reaction mixture, conc. NH4Cl solution, the reaction mixture was extracted with EtOAc and the combined organic phases were washed with brine, dried, concentrated, triturated with MeOH and filtered to give 21 g (92%) of the expected product.

1H NMR (400 MHz, CDCl ₃ ) δ ppm 7.44 (s, 1H), 3.77 (s, 2H), 2.30 (s, 3H), 1.99 (bs, 3H), 1.74/1.52 (m, 12H).

Step C: 1-(1-adamantylmethyl)-5-methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyrazole.

To 21 g of the product from step B (58.95 mmol, 1 eq.) in 300 ml THF at -78°C was added 28.3 ml of 2.5 M butyllithium solution (70.8 mmol, 1.2 eq.), and the mixture was stirred at -78°C for 0.5 h, treatments

- 78 046336 add 16.4 g of 2-isopropoxy-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (88.1 mmol, 1.5 eq.) (added in portions after 40 min), and kept at -78°C for 24 hours. After quenching the reaction mixture, conc. solution of NH ₄ Cl at RT, the reaction mixture was extracted with EtOAc and the combined organic phases were washed with brine, dried, concentrated, triturated with MeOH, and filtered to obtain 19.7 g (94%) of the expected product.

1H NMR (500 MHz, DMSO^b) δ ppm 7.45 (s, 1H), 3.69 (s, 2H), 2.36 (s, 3H), 1.91 (m, 1H), 1.64/1.54 (m, 6H), 1.50 (m, 6H), 1.24 (s, 12H) ; ¹³ C NMR (500 MHz, DMSO^) δ ppm 146.9, 144.1, 104.6, 59.7, 40.6, 36.8, 35.4, 28.1, 25.1, 12.1; MSHR-ERI (m/z): [M+H]+, calculated for C ₂₁ H ₃₄ BN ₂ O ₂ : 357.2713; found: 357.2704.

Synthesis 10b: 1-{[1-(3-Methoxypropyl)cyclooctyl]methyl}-5-methyl-4-(4,4,5,5-tetramethyl-1,3,2dioxaborolan-2-yl)-1H-pyrazole .

Step A: Methyl 1-(3-methoxypropyl)cyclooctanecarboxylate.

To 4.74 g (1.14 eq.) of diisopropylamine in 90 ml of tetrahydrofuran at -78°C, 18.8 ml (1.14 eq.) of a 2.5 M solution of butyllithium was added and, after incubating for 0.5 h at -78°C, 7.0 g (41.1 mmol) was added ) methyl cyclooctane carboxylate in 40 ml tetrahydrofuran for 1 hour. After standing for 1 hour at -78°C, 7.2 g (1.14 eq.) 1-bromo-3-methoxypropane was added and the mixture was stirred for 18 hours. After quenching reaction mixture by adding saturated NH4Cl solution, the mixture was extracted with EtOAc and the organic phases were dried over MgSO ₄ and concentrated to obtain 8.0 g (80%) of the expected product.

1H NMR (400 MHz, CDCl ₃ ) δ ppm 3.66 (s, 3H), 3.33 (t, 2H), 3.31 (s, 3H), 2.03-1.94 (m, 2H), 1.64-1.38 (m, 16H).

Step B: [1-(3-methoxypropyl)cyclooctyl]methanol.

To 9.0 g (37.13 mmol) of the product from step A in 93 ml of diethyl ether at 0°C, 1.76 g (1.25 eq.) of lithium aluminum hydride was added in portions. After stirring at room temperature. for 2 hours, the reaction mixture was quenched by adding ice water and EtOAc, and 10% NaOH solution was added. The mixture was extracted with EtOAc, dried and concentrated to give 7.4 g (93%) of the expected product.

1H NMR (400 MHz, CDCl ₃ ) δ ppm 3.37 (t, 2H), 3.34 (s, 3H), 3.30 (s, 2H), 1.61-1.23 (m, 18H).

Step C: 4-iodo-1-[[1-(3-methoxypropyl)cyclooctyl]methyl]pyrazole.

To 1.39 g (6.5 mmol) of the product from step B and 1.64 g (1.3 eq.) 4-iodo-1H-pyrazole in 33 ml of tetrahydrofuran were added 2.22 g (1.3 eq.) triphenylphosphine and 1.95 g (1.3 eq.) di-tert -butyl azodicarboxylate, and the mixture was stirred at room temperature. for 67 hours. 278 mg of 4-iodo-1H-pyrazole, 444 mg of triphenylphosphine, and 390 mg of di-tert-butyl azodicarboxylate were added to the mixture and stirred at room temperature. for 24 hours. After adding the reagents and stirring at room temperature. repeated for 24 hours (115 hours of stirring in total), the mixture was concentrated and purified by flash column chromatography (silica gel) using heptane and EtOAc as eluents to obtain 1.24 g (49%) of the desired product.

1H NMR (400 MHz, CDCl ₃ ) δ ppm 7.47 (s, 1H), 7.42 (s, 1H), 3.93 (s, 2H), 3.37 (t, 2H), 3.36 (s, 3H), 1.681.18 (m, 18H).

Step D: 4-iodo-1-[[1-(3-methoxypropyl)cyclooctyl]methyl]-5-methylpyrazole.

To 1.2 g (3.07 mmol) of the product from step C in 5 ml of tetrahydrofuran at -78°C was added 3.7 ml (1.2 eq.) 1 M LDA solution. After 0.6 h at -78°C, 0.5 ml (1.14 eq.) of methyl iodide was added dropwise to the mixture and the mixture was allowed to warm to room temperature. for 20 hours. The reaction mixture was quenched with saturated NH4Cl solution and extracted with EtOAc. The combined organic phases were dried, concentrated and purified by flash column chromatography (silica gel) using heptane and EtOAc as eluents to give 0.79 g (64%) of the expected product.

1H NMR (400 MHz, CDCl ₃ ) δ ppm 7.43 (s, 1H), 3.85 (s, 2H), 3.38 (t, 2H), 3.35 (s, 3H), 2.29 (s, 3H), 1.691.24 (m, 18H).

Step E: 1-[[1-(3-methoxypropyl)cyclooctyl]methyl]-5-methyl-4-(4,4,5,5-tetramethyl-1,3,2dioxaborolan-2-yl)pyrazole.

To a solution of 0.81 g (2 mmol) of the product from step D in 15 ml of tetrahydrofuran at -78°C, 0.96 ml (1.2 eq.) of a 2.5 M solution of butyllithium was added dropwise. After 0.5 h, 0.5 ml (1.2 eq.) of 2-isopropoxy-4,4,5,5-tetramethyl-1,3,2-dioxaborolane was added over 20 min, and the mixture was kept at -78°C for 6 h and at k.t. also for 6 hours. After quenching the reaction mixture with saturated NH4Cl solution and extraction with EtOAc, the combined organic phases were washed with brine, dried and purified by flash column chromatography (silica gel) using heptane and EtOAc as eluents to obtain 0.33 g ( 34%) of the target product.

1H NMR (500 MHz, DMSOHO δ ppm 7.46 (s, 1H), 3.75 (s, 2H), 3.27 (t, 2H), 3.21 (s, 3H), 2.36 (s, 3H), 1.66-1.1 (m, 14H), 1.57 (m, 2H), 1.24 (s, 12H), 1.24 (m, 2H) ¹³ C NMR (500 MHz, DMSO-d^ δ ppm 147.3, 144.5, 104.5, 73.2 , 58.2, 54.4, 40.5, 33.2, 25.1, 23.6, 11.8 IR: 2922, 1556, 1246, 1144, 1055 HRMS-ERI (m/z): [M+H]+, calculated for C ₂₃ H ₄₂ N ₂ O ₃ B: 405.3289; found: 404.3334.

Synthesis of 10c: 1-{[1-(3-methoxypropyl)cyclohexyl]methyl}-5-methyl-4-(4,4,5,5-tetramethyl-1,3,2dioxaborolan-2-yl)-1H-pyrazole .

- 79 046336

Step A: Methyl 1-(3-methoxypropyl)cyclohexanecarboxylate.

To 6.84 g (1.09 eq.) of diisopropylamine in 130 ml of tetrahydrofuran at -78°C, 27 ml (1.09 eq.) of a 2.5 M solution of butyllithium was added, and after 0.5 h at -78°C, 8.8 g of methyl cyclohexane carboxylate was added in 50 ml of tetrahydrofuran in for 1 hour. After standing for 1 hour at -78°C, 10.7 g (1.13 eq.) 1-bromo-3-methoxypropane was added and the mixture was stirred for 18 hours. After quenching the reaction mixture by adding a saturated solution of NH4C1, the mixture was extracted with EtOAc and the organic phases were dried over MgSO ₄ and concentrated to give 12 g (92%) of the expected product.

¹ H NMR (400 MHz, CDCl ₃ ) δ ppm 3.67 (s, 3H), 3.35 (d, 1H), 3.32 (d, 1H), 3.31 (s, 3H), 2.11-2.03 (m, 2H), 1.60-1.16 (m, 12H).

Step B: [1-(3-methoxypropyl)cyclohexyl]methanol.

To 12 g (56.41 mmol) of the product from step A in 140 ml of diethyl ether at 0°C, 2.68 g (1.25 eq.) of lithium aluminum hydride was added in portions. After stirring at room temperature. for 2 hours, the reaction mixture was quenched by adding ice water, and EtOAc and 10% NaOH solution were added. The mixture was extracted with EtOAc, dried and concentrated to give 9.37 g (89%) of the expected product.

1H NMR (400 MHz, CDCl ₃ ) δ ppm 3.41 (s, 2H), 3.38 (t, 2H), 3.35 (s, 3H), 1.56-1.27 (m, 14H).

Step C: 4-iodo-1-[[1-(3-methoxypropyl)cyclohexyl]methyl]pyrazole.

To 1.21 g (6.5 mmol) of the product from step B and 2.58 g (2.05 eq.) of 4-iodo-1H-pyrazole in 33 ml of tetrahydrofuran, 3.5 g (2.05 eq.) of triphenylphosphine and 3.07 g (2.05 eq.) of di-tert were added -butyl azodicarboxylate, and the mixture was stirred at room temperature. for 2 hours. 140 mg of 4-iodo-1H-pyrazole, 230 mg of triphenylphosphine and 200 mg of di-tert-butyl azodicarboxylate were added to the mixture and stirred at room temperature. for 24 hours. After adding the reagents and stirring at room temperature. within 24 hours was repeated twice (96 hours of stirring in total), the mixture was concentrated and purified by flash column chromatography (silica gel) using heptane and EtOAc as eluents to obtain 1.4 g (59.5%) of the desired product.

1H NMR (400 MHz, CDCl ₃ ) δ ppm 7.47 (s, 1H), 7.41 (s, 1H), 4.00 (s, 2H), 3.36 (t, 2H), 3.35 (s, 3H), 1.621.21 (m, 14H).

Step D: 4-iodo-1-[[1-(3-methoxypropyl)cyclohexyl]methyl]-5-methylpyrazole.

To 3.7 g (10.21 mmol) of the product from step C in 15 ml of tetrahydrofuran at -78°C was added 12.3 ml (1.2 eq.) of a 1 M solution of LDA in tetrahydrofuran. After 0.6 h at -78°C, 0.73 mL (1.14 eq) of methyl iodide was added dropwise to the mixture and the mixture was allowed to warm to room temperature. for 20 hours. The reaction mixture was quenched with saturated NH ₄ Cl solution and extracted with EtOAc. The combined organic phases were dried, concentrated and purified by flash column chromatography (silica gel) using heptane and EtOAc as eluents to give 2.85 g (74%) of the expected product.

1H NMR (400 MHz, CDCl ₃ ) δ ppm 7.44 (s, 1H), 3.92 (s, 2H), 3.38 (t, 2H), 3.35 (s, 3H), 2.29 (s, 3H), 1.581.13 (m, 14H).

Step E: 1-[[1-(3-methoxypropyl)cyclohexyl]methyl]-5-methyl-4-(4,4,5,5-tetramethyl-1,3,2dioxaborolan-2-yl)pyrazole.

To a solution of 5.0 g (13.3 mmol) of the product from step D in 71 ml of tetrahydrofuran at -78°C, 6.38 ml (1.2 eq.) of a 2.5 M solution of butyllithium was added dropwise. After 0.5 h, 4.1 ml (1.5 eq.) of 2isopropoxy-4,4,5,5-tetramethyl-1,3,2-dioxaborolane was added over 40 min, and the mixture was kept at -78°C for 6 h and at .T. for 6 hours. After quenching the reaction mixture with saturated NH4Cl solution and extraction with EtOAc, the combined organic phases were washed with brine, dried and purified by flash column chromatography (silica gel) using heptane and EtOAc as eluents to give 2.3 g ( 46%) of the target product.

1H NMR (500 MHz, DMSOHO δ ppm 7.47 (s, 1H), 3.84 (s, 2H), 3.27 (t, 2H), 3.2 (s, 3H), 2.37 (s, 3H), 1.54-1.07 (m, 10H), 1.46 (m, 2H), 1.32 (m, 2H), 1.24 (s, 12H) ¹³ C NMR (500 MHz, DMSOO δ ppm 147.3, 144.4, 104.6, 73.1, 58.2, 55.7, 37.9, 30.6, 25.1, 23.1, 12.0. IR: 2927, 1556, 1257, 1144, 1053. HRMS-ERI (m/z): [M+H]+, calculated for C ₂₁ H ₃₈ N ₂ O ₃ B: 376.2897; found: 376.3019.

Synthesis 11a: ethyl 5-bromo-2-[(4,5-dimethyl-6-{[(22)-3-{[2-(trimethylsilyl)ethoxy]methyl}-2,3-dihydro1,3-benzothiazol- 2-ylidene]amino}pyridazin-3-yl)amino] - 1,3-thiazol-4-carboxylate.

Step A: (22)-N(6-chloro-4,5-dimethylpyridazin-3-yl)-3-{[2-(trimethylsilyl)ethoxy]methyl}-2,3dihydro-1,3-benzothiazol-2- imin.

N.N-Diisopropylethylamine (3.6 ml, 20.6 mmol, 2 equiv.) was added under nitrogen atmosphere to a suspension of synthesis product 9a (3 g, 10.3 mmol, 1 equiv.) in dichloromethane (75 ml) at 0°C. [2(chloromethoxy)ethyl]trimethylsilane (2 mL, 11.3 mmol, 1.1 eq.) and 4-dimethylaminopyridine (63.0 mg, 0.52 mmol, 0.05 eq.) were added and the mixture was stirred at room temperature for 3 h. Dichloromethane ( 75 ml), and the mixture was washed with water (2x75 ml) and brine (75 ml). The solution was dried (magnesium sulfate) and concentrated in vacuo. Purification by flash silica gel Isolute column chromatography (20 g) eluting with dichloromethane provided the title product as a pale brown solid (3.95 g, 8.44 mmol, 82%), which was used directly in the next step without further definition of characteristics.

Step B: ethyl 2-[(4,5-dimethyl-6-{[(22)-3-{[2-(trimethylsilyl)ethoxy]methyl}-2,3-dihydro-1,3benzothiazol-2-ylidene] amino}pyridazin-3-yl)amino]-1,3-thiazol-4-carboxylate.

The product from step A (500 mg, 1.19 mmol, 1 eq.) and ethyl 2-amino-1,3-thiazol-4-carboxylate (245.4 mg, 1.43 mmol, 1.2 eq.) were added to the cesium carbonate solution under nitrogen atmosphere ( 1.16 g, 3.56 mmol, 3 eq.), tris(dibenzylideneacetone)dipalladium(0) (54.4 mg, 0.06 mmol, 0.05 eq.) and xanthos (68.7 mg, 0.12 mmol, 0.1 eq.) in alpha, alpha, alpha- trifluorotoluene (10 ml). The mixture was bubbled with nitrogen (10 min) and then heated at 170°C for 1 h under microwave irradiation. The solution was allowed to cool to ambient temperature and concentrated in vacuo. The mixture was partitioned between ethyl acetate and water, and the organic phase was washed with brine, dried (magnesium sulfate) and the solvent was removed in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 24 g RediSep™ silica gel cartridge) eluting with a gradient of 0-50% ethyl acetate in isoheptane gave the title product as an orange foam (301 mg, 0.54 mmol, 46%).

LC/MS (C ₂₅ H ₃₂ N ₆ O ₃ SiS ₂ ) 557 [M+H]+; RT 1.49 (ZHMS-U-B1).

1H NMR (400 MHz, DMSOHO δ 10.98 (s, 1H), 7.93 (s, 1H), 7.81 (dd, J=7.6, 1.1 Hz, 1H), 7.48-7.37 (m, 2H), 7.27-7.20 (m , 1H), 5.84 (s, 2H), 4.29 (q, J=7.1 Hz, 2H), 3.71 (dd, J=8.4, 7.5 Hz, 2H), 2.41 (s, 3H), 2.35 (s, 3H) , 1.31 (t, J=7.1 Hz, 3H), 0.97-0.86 (m, 2H), -0.12 (s, 9H).

Step C: ethyl 5-bromo-2-[(4,5-dimethyl-6-{[(22)-3-{[2-(trimethylsilyl)ethoxy]methyl}-2,3-dihydro1,3-benzothiazol- 2-ylidene]amino}pyridazin-3-yl)amino] - 1,3-thiazol-4-carboxylate.

N-Bromosuccinimide (125 mg, 0.7 mmol, 1.3 eq.) was added to a stirred solution of the product from Step B (301 mg, 0.54 mmol, 1 eq.) in dichloromethane (15 ml). The reaction mixture was stirred at ambient temperature for 1.5 h, then the solvent was removed in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 24 g RediSep™ silica gel cartridge) eluting with a gradient of 0-60% ethyl acetate in isoheptane gave the title product as a brown gum (198 mg, 0.31 mmol, 58%).

LC/MS (C ₂ 5H ₃₁ BrN ₆ O ₃ SiS ₂ ) 637 [M+H]+; RT 1.57 (ZHMS-U-B1).

1H NMR (400 MHz, DMSO-de) δ 11.31 (s, 1H), 7.82 (d, 1H), 7.48-7.38 (m, 2H), 7.28-7.20 (m, 1H), 5.84 (s, 2H), 4.30 (q, J=7.1 Hz, 2H), 3.70 (dd, J=8.5, 7.4 Hz, 2H), 2.41 (s, 3H), 2.34 (s, 3H), 1.32 (t, J=7.1 Hz, 3H ), 0.97-0.86 (m, 2H), -0.12 (s, 9H).

Synthesis 11b: ethyl 5-bromo-2-[methyl(5-methyl-6-{[(22)-3-{[2-(trimethylsilyl)ethoxy]methyl}-2,3dihydro-1,3-benzothiazol-2 -ylidene]amino}pyridazin-3 -yl)amino] - 1,3-thiazol-4-carboxylate.

Step A: ethyl 2-[(6-chloro-5-methylpyridazin-3-yl)(methyl)amino]-1,3-thiazol-4-carboxylate.

A solution of 3,6-dichloro-4-methylpyridazine (662 mg, 4.06 mmol, 1.02 eq.) and ethyl 2-(methylamino)-1,3thiazol-4-carboxylate (742 mg, 3.98 mmol, 1 eq.) in tetrahydrofuran ( 20 mL) was cooled in an ice bath, then sodium hydride (60% in mineral oil; 188 mg, 4.7 mmol, 1.18 eq.) was slowly added. The mixture was kept in an ice bath for 1 hour, then stirred at ambient temperature for 3 hours. A saturated aqueous solution of ammonium chloride was added and the resulting solution was extracted with dichloromethane (x2). The combined organic extracts were washed with water (x2) followed by brine, then dried (magnesium sulfate) and the solvent was removed in vacuo. The residue was triturated with dichloromethane and methanol, and the solids were collected by filtration and dried in vacuo to give the title product as a cream solid (324 mg, 1.04 mmol, 26%).

LC/MS (C ₁₂ H ₁₃ ClN ₄ O ₂ S) 313 [M+H]+; RT 1.15 (ZHMS-U-B1).

1H NMR (400 MHz, DMSO<) δ 8.04 (s, 1H), 7.87 (d, J=1.2 Hz, 1H), 4.30 (q, J=7.1 Hz, 2H), 3.83 (s, 3H), 2.44 ( d, J=1.0 Hz, 3H), 1.32 (t, J=7.1 Hz, 3H).

Step B: ethyl 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)-1,3thiazol-4-carboxylate.

The product from step A (467 mg, 1.49 mmol, 1 eq.) and 2-aminobenzothiazole (269 mg, 1.79 mmol, 1.2 eq.) were added to a mixture of cesium carbonate (1.46 g, 4.47 mmol, 3 eq.) under nitrogen atmosphere. tris(dibenzylideneacetone)dipalladium(0) (68.3 mg, 0.07 mmol, 0.05 eq.) and xantphos (86.3 mg, 0.15 mmol, 0.1 eq.) in alpha,alpha,alpha-trifluorotoluene (10 ml). The mixture was bubbled with nitrogen (10 min) and then heated at 170°C for 1 h under microwave irradiation. The solution was allowed to cool to ambient temperature, filtered, eluting with dichloromethane, and the filtrate was concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 24 g RediSep™ silica gel cartridge) eluting with a gradient of 0-5% methanol in dichloromethane provided the title product as a yellow solid (133 mg, 0.31 mmol, 21%) .

LC/MS (C ₁₉ H ₁₈ N ₆ O ₂ S ₂ ) 427 [M+H]+; RT 1.30 (ZHMS-U-B1).

1H NMR (400 MHz, DMSOO δ 11.10 (s, 1H), 7.99 (d, J=6.9 Hz, 1H), 7.76-7.67 (m, 1H), 7.54-7.34 (m, 2H), 7.28-7.17 (m , 1H), 4.30 (q, J=7.2 Hz, 2H), 3.83 (s, 3H), 2.48 (s, 3H), 1.31 (t, 3H).

Step C: ethyl 2-[methyl(5-methyl-6-{[(22)-3-{[2-(trimethylsilyl)ethoxy]methyl}-2,3-dihydro-1,3benzothiazol-2-ylidene]amino }pyridazin-3-yl)amino]-1,3-thiazol-4-carboxylate.

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A solution of the product from step B (133 mg, 0.31 mmol, 1 eq.) in dichloromethane (5 ml) was cooled in an ice bath, then N.N-diisopropylethylamine (0.1 ml, 0.62 mmol, 2 eq.) was added. After 5 min, 4-dimethylaminopyridine (1.9 mg, 0.02 mmol, 0.05 eq.) was added followed by 2(trimethylsilyl)ethoxymethyl chloride (0.06 mL, 0.34 mmol, 1.1 eq.). The mixture was allowed to warm slowly to ambient temperature and stirred overnight. The reaction mixture was diluted with dichloromethane, washed successively with water (x2) and brine, dried (magnesium sulfate), and the solvent was removed in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 12 g RediSep™ silica gel cartridge) eluting with a gradient of 0-50% ethyl acetate in isoheptane gave the title product as a yellow glass (56.3 mg, 0.1 mmol, 32%).

LC/MS (C ₂₅ H ₃₂ N ₆ O ₃ SiS ₂ ) 557 [M+H]+; RT 1.64 (ZHMS-U-B1).

1H NMR (400 MHz, DMSO^ ₆ ) δ 7.97 (s, 1H), 7.85 (dd, J=7.6, 1.0 Hz, 1H), 7.71 (d, J=1.1 Hz, 1H), 7.507.40 (m, 2H), 7.29-7.22 (m, 1H), 5.87 (s, 2H), 4.29 (q, J=7.1 Hz, 2H), 3.83 (s, 3H), 3.78-3.67 (m, 2H), 2.46 (d , J=1.0 Hz, 3H), 1.31 (t, J=7.1 Hz, 3H), 0.98-0.87 (m, 2H), -0.12 (s, 9H).

Step D: ethyl 5-bromo-2-[methyl(5-methyl-6-{[(22)-3-{[2-(trimethylsilyl)ethoxy]methyl}-2,3dihydro-1,3-benzothiazol-2 -ylidene]amino}pyridazin-3 -yl)amino] - 1,3-thiazol-4-carboxylate.

N-Bromosuccinimide (23.4 mg, 0.13 mmol, 1.3 eq.) was added to a stirred solution of the product from step C (56.3 mg, 0.1 mmol, 1 eq.) in dichloromethane (5 ml). The reaction mixture was stirred at ambient temperature for 1.5 h, then the solvent was removed in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 24 g RediSep™ silica gel cartridge) eluting with a gradient of 0-60% ethyl acetate in isoheptane provided the title product as a white solid (54.7 mg, 0.09 mmol, 85%) .

LC/MS (C ₂₅ H ₃₁ BrN ₆ O ₃ SiS ₂ ) 635 [M+H]+; RT 1.73 (ZHMS-U-B1).

1H NMR (400 MHz, DMSO-a6) δ 7.86 (dd, J=7.7, 1.0 Hz, 1H), 7.73 (d, J=1.1 Hz, 1H), 7.52-7.38 (m, 2H), 7.25 (ddd, J=8.3, 7.0, 1.5 Hz, 1H), 5.87 (s, 2H), 4.31 (q, J=7.1 Hz, 2H), 3.78 (s, 3H), 3.76-3.66 (m, 2H), 2.46 (d , J=1.0 Hz, 3H), 1.32 (t, J=7.1 Hz, 3H), 0.98-0.85 (m, 2H), -0.11 (s, 9H).

Synthesis 11с: ethyl 5-bromo-2-[(3-methoxypropyl)(5-methyl-6-{[(22)-3-{[2(trimethylsilyl)ethoxy]methyl}-2,3-dihydro-1, 3-benzothiazol-2-ylidene]amino}pyridazin-3-yl)amino]1,3-thiazol-4-carboxylate.

Step A: ethyl 2-[(3-methoxypropyl)amino]-1,3-thiazole-4-carboxylate.

Ammonia (2 M in isopropanol; 60 ml, 0.12 mol, 3.45 eq.) at 0°C was added to a solution of 3methoxypropyl isothiocyanate (4.5 g, 34.3 mmol, 1 eq.) in isopropanol (15 ml), and the mixture was stirred for 1 h at 0°C and then for 18 hours at ambient temperature. The suspension was concentrated in vacuo to give a pale yellow resin. Ethanol (60 mL) and ethyl bromopyruvate (5 mL, 35.9 mmol, 1.05 eq.) were added and the mixture was stirred for 1 h under nitrogen atmosphere. Triethylamine (10 mL, 68.6 mmol, 2 eq.) was added and the mixture was stirred at ambient temperature for 18 h. The reaction mixture was partitioned between ethyl acetate (300 mL) and water, and the organic phase was washed successively with water (150 mL) and brine. solution (100 ml), dried (magnesium sulfate) and concentrated in vacuo. Purification by flash column chromatography (100 g silica gel) eluting with 99:1 dichloromethane/methanol gave a dark yellow oil which was triturated with heptane (30 ml), filtered, washed with heptane (20 ml) and dried in vacuo to obtain the target product in the form of a pale brown powder (5.75 g, 23.5 mmol, 69%).

LC/MS (C ₁₀ H ₁₆ N ₂ O ₃ S) 245 [M+H]+; RT 0.99 (ZHMS-U-B1).

1H NMR (400 MHz, CDC13) δ 7.38 (d, J=0.7 Hz, 1H), 6.03 (t, J=5.6 Hz, 1H), 4.34 (q, J=7.1 Hz, 2H), 3.543.42 (m , 2H), 3.40-3.33 (m, 2H), 3.32 (s, 3H), 1.97-1.83 (m, 2H), 1.36 (t, J=7.1 Hz, 3H).

Step B: ethyl 2-[(6-chloro-5-methylpyridazin-3-yl)(3-methoxypropyl)amino]-1,3-thiazol-4carboxylate.

Sodium hydride (60% dispersion; 350 mg, 8.75 mmol, 1.22 eq.) under nitrogen was added to a solution of the product from step A (1.75 g, 7.16 mmol, 1 eq.) and 3,6-dichloro-4-methylpyridazine (1.3 g, 7.98 mmol, 1.11 eq.) in 1,4-dioxane (30 ml) at 0°C, the mixture was stirred for 1 hour at 0°C and then heated at 60°C for 6 hours. The reaction mixture was allowed to cool to ambient temperature, then partitioned between ethyl acetate (300 ml) and saturated aqueous ammonium chloride (150 ml). The organic phase was washed successively with water (2x150 ml) and brine (100 ml), dried (magnesium sulfate) and concentrated in vacuo. Purification by flash column chromatography (50 g silica gel) eluting with 39:1 dichloromethane/methanol gave a brown oil which was triturated with diethyl ether (10 ml), filtered, washed with diethyl ether (10 ml) and dried in vacuo to give the target product in the form of a pale brown powder (1.3 g, 3.51 mmol, 49%).

LC/MS (C ₁₅ H ₁₉ ClN ₄ O ₃ S) 371 [M+H]+; RT 1.28 (ZHMS-U-B1).

1H NMR (400 MHz, CDC13) δ 7.81 (s, 1H), 7.55 (d, J=1.1 Hz, 1H), 4.57-4.44 (m, 2H), 4.37 (q, J=7.1 Hz, 2H), 3.45 (t, J=5.5 Hz, 2H), 3.34 (s, 3H), 2.45 (d, J=0.9 Hz, 3H), 2.20-2.06 (m, 2H), 1.39 (t, J=7.1 Hz, 3H) .

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Step C: ethyl 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(3methoxypropyl)amino)-1,3-thiazol-4-carboxylate.

The product from step B (1.3 g, 3.51 mmol, 1 eq.) and 2-aminobenzothiazole (600 mg, 3.99 mmol, 1.14 eq.) were added to a solution of tris(dibenzylideneacetone)dipalladium(0) (161 mg, 0.18 mmol) under a nitrogen atmosphere , 0.05 eq.) and xanthos (203 mg, 0.35 mmol, 0.1 eq.) in 1,4-dioxane (20 ml), and the mixture was stirred. N,N-diisopropylethylamine (1.9 mL, 10.5 mmol, 3 eq.) was added and the mixture was heated in a sealed tube at 150°C for 24 hours. The solution was allowed to cool to ambient temperature and concentrated in vacuo. Purification by flash column chromatography (50 g silica gel) eluting with 99:1 dichloromethane/methanol gave a red gum which was triturated with methanol (15 ml), filtered, washed with methanol (10 ml) and dried in vacuo to give the target product as a yellow solid (1.35 g, 2.79 mmol, 80%).

LC/MS (C ₂₂ H ₂₄ N6O3S2) 485 [M+H]+; RT 1.41 (ZHMS-U-B1).

1H NMR (400 MHz, DMSO<) δ 11.15 (br s, 1H), 7.97 (s, 1H), 7.92 (br s, 1H), 7.69 (s, 1H), 7.56 (br s, 1H), 7.40 ( t, J=7.4 Hz, 1H), 7.22 (t, J=7.5 Hz, 1H), 4.44 (t, J=7.2 Hz, 2H), 4.29 (q, J=7.1 Hz, 2H), 3.43 (t, J=6.0 Hz, 2H), 3.25 (s, 3H), 2.48 (s, 3H), 2.05-1.93 (m, 2H), 1.32 (t, J=7.1 Hz, 3H).

Step D: ethyl 2-[(3-methoxypropyl)(5-methyl-6-{[(22)-3-{[2-(trimethylsilyl)ethoxy]methyl}-2,3dihydro-1,3-benzothiazol-2 -ylidene]amino}pyridazin-3 -yl)amino] - 1,3-thiazol-4-carboxylate.

A solution of the product from step C (860 mg, 1.77 mmol, 1 eq.) in dichloromethane (30 mL) was cooled in an ice bath, then N,N-diisopropylethylamine (0.59 mL, 3.55 mmol, 2 eq.) was added. After stirring for 10 min, 2-(trimethylsilyl)ethoxymethyl chloride (0.37 mL, 2.13 mmol, 1.2 eq.) was added followed by 4-(dimethylamino)pyridine (10.8 mg, 0.09 mmol, 0.05 eq.). The reaction mixture was allowed to warm to ambient temperature and stirred for 5 hours. The mixture was diluted with dichloromethane, then washed with water followed by brine, dried (magnesium sulfate) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 24 g RediSep™ silica gel cartridge) eluting with a gradient of 0-50% ethyl acetate in isoheptane gave the title product as a creamy foam (0.83 g, 1.35 mmol, 76%).

LC/MS (C28H38N6O4SiS2) 615 [M+H]+; RT 1.51 (ZHMS-U-B1).

1H NMR (400 MHz, DMSO-b6) δ 7.96 (s, 1H), 7.85 (d, 1H), 7.69 (s, 1H), 7.50-7.40 (m, 2H), 7.30-7.22 (m, 1H), 5.86 (s, 2H), 4.43 (t, J=7.2 Hz, 2H), 4.28 (q, J=7.1 Hz, 2H), 3.78-3.67 (m, 2H), 3.42 (t, J=6.0 Hz, 2H ), 3.24 (s, 3H), 2.46 (d, J=0.9 Hz, 3H), 2.06-1.92 (m, 2H), 1.31 (t, J=7.1 Hz, 3H), 0.98-0.86 (m, 2H) , -0.12 (s, 9H).

Step E: ethyl 5-bromo-2-[(3-methoxypropyl)(5-methyl-6-{[(22)-3-{[2(trimethylsilyl)ethoxy]methyl}-2,3-dihydro-1, 3-benzothiazol-2-ylidene]amino}pyridazin-3-yl)amino]1,3-thiazol-4-carboxylate.

N-Bromosuccinimide (63.7 mg, 0.36 mmol, 1.1 eq.) was added to a stirred solution of the product from Step A (200 mg, 0.33 mmol, 1 eq.) in dichloromethane (10 ml), and the mixture was stirred for 4 hours. The solvent was removed in vacuo and the crude product was purified by automated flash column chromatography (CombiFlash Rf, 12 g RediSep™ silica gel cartridge) eluting with a gradient of 0-60% ethyl acetate in isoheptane to give the title product as a creamy foam (193 mg, 0.28 mmol, 85%).

LC/MS (C ₂₈ H ₃₇ BrN ₆ O ₄ SiS ₂ ) 695 [M+H]+; RT 1.60 (ZHMS-U-B1).

1H NMR (400 MHz, DMSO-b6) δ 7.87 (dd, J=7.6, 1.1 Hz, 1H), 7.72 (d, J=1.2 Hz, 1H), 7.53-7.39 (m, 2H), 7.26 (ddd, J=8.3, 7.0, 1.4 Hz, 1H), 5.87 (s, 2H), 4.43 (t, J=7.1 Hz, 2H), 4.31 (q, J=7.1 Hz, 2H), 3.77-3.67 (m, 2H ), 3.41 (t, J=6.0 Hz, 2H), 3.23 (s, 3H), 2.47 (s, 3H), 2.04-1.90 (m, 2H), 1.32 (t, J=7.1 Hz, 3H), 0.97 -0.86 (m, 2H), -0.11 (s, 9H).

Synthesis 11d: ethyl 5-bromo-2-[(5-cyclopropyl-6-{[(22)-3-{[2-(trimethylsilyl)ethoxy]methyl}-2,3dihydro-1,3-benzothiazol-2- ylidene]amino}pyridazin-3-yl)(methyl)amino]-1,3-thiazol-4-carboxylate.

Step A: ethyl 2-[(6-chloro-5-cyclopropylpyridazin-3-yl)(methyl)amino]-1,3-thiazol-4-carboxylate.

To a solution of synthesis product 71 (495 mg, 2.62 mmol, 1 eq.) and ethyl 2-(methylamino)-1,3-thiazol-4carboxylate (488 mg, 2.62 mmol, 1 eq.) in tetrahydrofuran (25 ml) in atmosphere nitrogen, sodium hydride (60% in mineral oil; 124 mg, 3.09 mmol, 1.18 eq.) was slowly added at 0°C. The mixture was stirred for 1 hour at 0°C, then allowed to warm to ambient temperature and stirred for 18 hours. Saturated aqueous ammonium chloride (75 ml) was added and the mixture was extracted with dichloromethane (250 ml). The organics were washed successively with water (2x75 ml) and brine (75 ml), dried (magnesium sulfate) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 40 g RediSep™ silica gel cartridge) eluting with a gradient of 0-60% ethyl acetate in isoheptane provided the title product as a white crystalline solid (398 mg, 1.17 mmol, 45% ).

LC/MS (C14H15C1N4O2S) 339 [M+H]+; RT 1.25 (ZHMS-U-B1).

1H NMR (400 MHz, DMSO-b6) δ 8.03 (s, 1H), 7.24 (s, 1H), 4.30 (q, J=7.1 Hz, 2H), 3.83 (s, 3H), 2.23-2.12 (m, 1H), 1.32 (t, J=7.1 Hz, 3H), 1.24-1.16 (m, 2H), 1.13-1.06 (m, 2H).

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Step B: ethyl 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-cyclopropylpyridazin-3-yl}(methyl)amino)1,3-thiazol-4-carboxylate.

2-Aminobenzothiazole (0.21 g, 1.41 mmol, 1.2 eq.) and the product from step A (398 mg, 1.17 mmol, 1 eq.) were added under nitrogen to a solution of xantphos (68 mg, 0.12 mmol, 0.1 eq.) and Tris (dibenzylideneacetone)dipalladium(0) (53.8 mg, 0.06 mmol, 0.05 eq.) in 1,4-dioxane (20 ml). N.N-Diisopropylethylamine (0.61 mL, 3.52 mmol, 3 eq.) was added and the mixture was heated in a sealed tube at 150°C for 24 h. The solution was allowed to cool to ambient temperature and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 40 g RediSep™ silica gel cartridge) eluting with a gradient of 0-90% ethyl acetate in isoheptane provided the title product as a yellow solid (249 mg, 0.55 mmol, 47%) .

LC/MS (C ₂ iH ₂₀ N ₆ O ₂ S ₂ ) 453 [M+H]+; RT 1.38 (ZHMS-U-B1).

1H NMR (400 MHz, DMSO^) δ 7.96 (s, 1H), 7.90 (s, 1H), 7.50 (br s, 1H), 7.39 (t, J=7.6 Hz, 1H), 7.297.08 (m, 2H), 4.30 (q, 3H), 3.82 (s, 3H), 1.32 (t, 3H), 1.21-1.07 (m, 2H), 1.06-0.95 (m, 2H).

Step C: ethyl 2-[(5-cyclopropyl-6-{[(22)-3-{[2-(trimethylsilyl)ethoxy]methyl}-2,3-dihydro-1,3benzothiazol-2-ylidene]amino} pyridazin-3-yl)(methyl)amino]-1,3-thiazol-4-carboxylate.

A solution of the product from step B (221 mg, 0.49 mmol, 1 eq.) in dichloromethane (10 ml) was cooled in an ice bath, then N,N-diisopropylethylamine (0.16 ml, 0.98 mmol, 2 eq.) was added and the mixture was stirred in for 10 minutes. 2-(trimethylsilyl)ethoxymethyl chloride (0.1 mL, 0.59 mmol, 1.2 eq.) was added followed by 4-(dimethylamino)pyridine (2.98 mg, 0.02 mmol, 0.05 eq.) and the mixture was stirred at ambient temperature for 7 h. The reaction mixture was partitioned between dichloromethane and water, and the organic phase was washed with water followed by brine, dried (magnesium sulfate) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 12 g RediSep™ silica gel cartridge) eluting with a gradient of 0-50% ethyl acetate in isoheptane gave the title product as a creamy foam (185 mg, 0.32 mmol, 65%).

LC/MS (C ₂₇ H ₃₄ N6O ₃ SiS ₂ ) 583 [M+H]+; RT 1.45 (ZHMS-U-B1).

1H NMR (400 MHz, DMSOO δ 7.95 (s, 1H), 7.84 (dd, J=7.9, 1.1 Hz, 1H), 7.51-7.40 (m, 2H), 7.29-7.21 (m, 1H), 7.11 (s , 1H), 5.88 (s, 2H), 4.29 (q, J=7.1 Hz, 2H), 3.82 (s, 3H), 3.77-3.65 (m, 2H), 2.73-2.61 (m, 1H), 1.31 ( t, J=7.1 Hz, 3H), 1.24-1.12 (m, 2H), 1.11-1.03 (m, 2H), 0.96-0.86 (m, 2H), -0.13 (s, 9H).

Step D: ethyl 5-bromo-2-[(5-cyclopropyl-6-{[(22)-3-{[2-(trimethylsilyl)ethoxy]methyl}-2,3dihydro-1,3-benzothiazol-2- ylidene]amino}pyridazin-3-yl)(methyl)amino]-1,3-thiazol-4-carboxylate.

N-Bromosuccinimide (62.2 mg, 0.35 mmol, 1.1 eq.) was added to a stirred solution of the product from step C (185 mg, 0.32 mmol, 1 eq.) in dichloromethane (10 ml). After 2.5 h, the solvent was removed in vacuo and the crude product was purified by automated flash column chromatography (CombiFlash Rf, 12 g RediSep™ silica gel cartridge) eluting with a gradient of 0-60% ethyl acetate in isoheptane to give the title product as a cream solid ( 149 mg, 0.23 mmol, 71%).

LC/MS (C ₂₇ H ₃₃ BrN6O ₃ SiS ₂ ) 663 [M+H]+; RT 1.52 (ZHMS-U-B1).

1H NMR (400 MHz, DMSO-de) δ 7.86 (dd, J=7.8, 1.1 Hz, 1H), 7.52-7.39 (m, 2H), 7.30-7.22 (m, 1H), 7.09 (s, 1H), 5.88 (s, 2H), 4.31 (q, J=7.1 Hz, 2H), 3.79 (s, 3H), 3.75-3.66 (m, 2H), 2.74-2.61 (m, 1H), 1.33 (t, J= 7.1 Hz, 3H), 1.22-1.13 (m, 2H), 1.13-1.05 (m, 2H), 0.95-0.84 (m, 2H), -0.13 (s, 9H).

Synthesis 11e: ethyl 5-iodo-2-[methyl(5-methyl-6-{[(22)-3-{[2-(trimethylsilyl)ethoxy]methyl}-2,3dihydro-1,3-benzothiazol-2 -ylidene]amino}pyridazin-3 -yl)amino] - 1,3-thiazol-4-carboxylate.

Step A: ethyl 2-(methylamino)-1,3-thiazole-4-carboxylate.

To a suspension of N-methylthiourea (5 g, 55.5 mmol, 1 eq.) in ethanol (80 mL) cooled in ice water, ethyl bromopyruvate (7.45 mL, 59.4 mmol, 1.07 eq.) was slowly added. After 10 min, the mixture was allowed to warm to ambient temperature and stirred for 1.5 h. Triethylamine (15.4 mL, 111 mmol, 2 eq.) was added and the mixture was stirred at ambient temperature for 18 h. The reaction mixture was concentrated in vacuo, the residue partitioned between ethyl acetate and water, the aqueous phase was extracted with ethyl acetate (x2) and the combined organic extracts were dried (magnesium sulfate) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 80 g RediSep™ silica gel cartridge) eluting with a gradient of 0-100% ethyl acetate in isoheptane gave the title product as a beige solid (5.26 g, 28.2 mmol, 51%).

LC/MS (C ₇ Hj ₀ N ₂ O ₂ S) 187 [M+H]+; RT 0.85 (ZHMS-U-B1).

1H NMR (400 MHz, DMSOO δ 7.73 (q, J=4.7 Hz, 1H), 7.51 (d, J=0.5 Hz, 1H), 4.22 (q, J=7.1 Hz, 2H), 2.83 (d, J= 4.8 Hz, 3H), 1.27 (t, J=7.1 Hz, 3H).

Step B: ethyl 2-[(6-chloro-5-methylpyridazin-3-yl)(methyl)amino]-1,3-thiazol-4-carboxylate.

A solution of 3,6-dichloro-4-methylpyridazine (4.6 g, 28.2 mmol, 1 eq.) and the product from step A (5.26 g, 28.2 mmol, 1 eq.) in tetrahydrofuran (100 ml) was cooled in an ice bath and added in portions sodium hydride (60% in mineral oil; 1.24 g, 31.1 mmol, 1.1 eq.). The mixture was allowed to gradually warm up

- 84 046336 to ambient temperature and stirred for 18 hours. Dichloromethane and saturated aqueous ammonium chloride were added and a precipitate was observed to form. The solid was collected by filtration, washed with water and then dried in vacuo at 40°C for 12 hours to give the title product as a cream solid (4.24 g, 13.6 mmol, 48%).

LC/MS (C ₁₂ H ₁₃ ClN ₄ O ₂ S) 313 [M+H]+; RT 1.19 (LCMS-V-BI).

1H NMR (400 MHz, DMSO<) δ 8.04 (s, 1H), 7.87 (d, J=1.1 Hz, 1H), 4.30 (q, J=7.1 Hz, 2H), 3.83 (s, 3H), 2.44 ( d, J=1.0 Hz, 3H), 1.32 (t, J=7.1 Hz, 3H).

Step C: ethyl 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)-1,3thiazol-4-carboxylate.

Into a sealed flask containing the product from step B (4.24 g, 13.6 mmol, 1 eq.), 2-aminobenzothiazole (2.44 g, 16.3 mmol, 1.2 eq.) and xantphos (0.78 g, 1.36 mmol, 0.1 eq.) at 1.4 -dioxane (250 ml) under a nitrogen atmosphere, tris(dibenzylideneacetone)dipalladium(0) (0.62 g, 0.68 mmol, 0.05 eq.) was added, followed by the addition of N,N-diisopropylethylamine (6.74 ml, 40.7 mmol, 3 eq.), and the vessel was sealed and heated at 150°C for 24 hours. The reaction mixture was allowed to cool to ambient temperature and purified using automated flash column chromatography (CombiFlash Rf, 120 g RediSep™ silica gel cartridge) eluting with a gradient of 0-100% ethyl acetate in isoheptane to give the title product as a beige solid (3.05 g, 7.14 mmol, 53%).

LC/MS (C ₁₉ H ₁₈ N ₆ O ₂ S ₂ ) 427 [M+H]+; RT 1.16 (LCMS-V-BI).

1H NMR (400 MHz, DMSO<) δ 7.97 (s, 1H), 7.95-7.82 (m, 1H), 7.71 (s, 1H), 7.55 (br s, 1H), 7.39 (t, J=7.5 Hz, 1H), 7.22 (t, J=7.5 Hz, 1H), 4.31 (q, 3H), 3.83 (s, 3H), 2.48 (s, 3H), 1.33 (t, 3H).

Step D: Ethyl 2-[methyl(5-methyl-6-{[(22)-3-{[2-(trimethylsilyl)ethoxy]methyl}-2,3-dihydro-1,3benzothiazol-2-ylidene]amino }pyridazin-3-yl)amino]-1,3-thiazol-4-carboxylate.

To a suspension of the product from step C (3.05 g, 7.15 mmol, 1 eq.) in dichloromethane (100 ml), cooled to 0°C, was added N,N-diisopropylethylamine (2.37 ml, 14.3 mmol, 2 eq.). After 10 min, 2-(trimethylsilyl)ethoxymethyl chloride (1.51 mL, 8.58 mmol, 1.2 eq.) was added followed by 4(dimethylamino)pyridine (43.7 mg, 0.36 mmol, 0.05 eq.) and the mixture was allowed to warm to ambient temperature and stirred for 6 hours. The reaction mixture was washed with water, followed by brine, then dried (magnesium sulfate) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 80 g RediSep™ silica gel cartridge) eluting with a gradient of 0-50% ethyl acetate in isoheptane provided the title product as a white solid (1.38 g, 2.47 mmol, 35%) .

LC/MS (C ₂₅ H ₃₂ N ₆ O ₃ SiS ₂ ) 557 [M+H]+; RT 1.65 (LCMS-V-BI).

1H NMR (400 MHz, DMSO<) δ 7.96 (s, 1H), 7.84 (dd, 1H), 7.70 (d, J=0.9 Hz, 1H), 7.51-7.39 (m, 2H), 7.25 (ddd, J =8.3, 6.9, 1.6 Hz, 1H), 5.86 (s, 2H), 4.28 (q, J=7.1 Hz, 2H), 3.83 (s, 3H), 3.77-3.66 (m, 2H), 2.46 (d, J=1.0 Hz, 3H), 1.32 (t, J=7.1 Hz, 3H), 0.99-0.879 (m, 2H), -0.10 (s, 9H).

Step E: ethyl 5-iodo-2-[methyl(5-methyl-6-{[(22)-3-{[2-(trimethylsilyl)ethoxy]methyl}-2,3-dihydro1,3-benzothiazol-2 -ylidene]amino}pyridazin-3 -yl)amino] - 1,3-thiazol-4-carboxylate.

N-Iodosuccinimide (0.05 ml, 2.25 g/ml, 0.54 mmol, 1.5 eq.) was added to a stirred solution of the product from step D (200 mg, 0.36 mmol, 1 eq.) in dichloromethane (6 ml), and the mixture was stirred at environment for 18 hours. The reaction mixture was washed successively with water and brine, dried (magnesium sulfate) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 12 g RediSep™ silica gel cartridge) eluting with a gradient of 0-60% ethyl acetate in isoheptane provided the title product as a cream solid (180 mg, 0.26 mmol, 73%) .

LC/MS (C ₂₅ H ₃₁ IN ₆ O ₃ SiS ₂ ) 683 [M+H]+; RT 1.73 (LCMS-V-BI).

1H NMR (400 MHz, DMSO<) δ 7.88 (dd, J=7.5, 1.0 Hz, 1H), 7.71 (s, 1H), 7.53-7.40 (m, 2H), 7.31-7.22 (m, 1H), 5.86 (s, 2H), 4.32 (q, J=7.1 Hz, 2H), 3.82 (s, 3H), 3.78-3.69 (m, 2H), 2.48 (d, J=1.0 Hz, 3H), 1.34 (t, J=7.1 Hz, 3H), 0.99-0.89 (m, 2H), -0.13 (s, 9H).

Synthesis 11f: ethyl 5-bromo-2-[(6-{[(22)-3-{[2-(trimethylsilyl)ethoxy]methyl}-2,3-dihydro-1,3benzothiazol-2-ylidene]amino} pyridazin-3-yl)amino]-1,3-thiazol-4-carboxylate.

Step A: ethyl 2-[(6-{[(22)-3-{[2-(trimethylsilyl)ethoxy]methyl}-2,3-dihydro-1,3-benzothiazol-2ylidene] amino } pyridazine-3 - yl)amino]-1,3-thiazole-4-carboxylate.

Synthesis product 4b (1 g, 2.54 mmol, 1 eq.) and ethyl 2-amino-1,3-thiazol-4-carboxylate (500 mg, 2.9 mmol, 1.14 eq.) were added to the tris(dibenzylideneacetone) solution under nitrogen atmosphere. dipalladium(0) (116.5 mg, 0.13 mmol, 0.05 eq.) and xanthos (147.2 mg, 0.25 mmol, 0.1 eq.) in 1,4-dioxane (20 ml). N.N-Diisopropylethylamine (1.5 mL, 7.63 mmol, 3 eq.) was added and the mixture was heated at 100°C for 18 hours. The solution was allowed to cool to ambient temperature and concentrated in vacuo. Purification by column chromatography (50 g silica gel) eluting with 50% ethyl acetate in heptane gave a dark yellow solid. Trituration with heptane (20 ml) resulted in a solid, which was collected by filtration, washed with heptane (2x10 ml) and dried in a bath.

- 85 046336 kuume to obtain the target product in the form of a yellow solid (995 mg, 1.88 mmol, 74%) [mixture of isomers].

LC/MS (C ₂₃ H ₂₈ N ₆ O ₃ SiS ₂ ) 529 [M+H]+; RT 1.54 (LCMS-V-BI).

1H NMR (400 MHz, Chloroform-d) δ 10.83 (s, 1H), 7.90-7.29 (m, 6H), 7.26-7.13 (m, 1H), 5.95-5.68 (m, 2H), 4.53-4.19 (m , 2H), 3.85-3.68 (m, 2H), 1.39-1.19 (m, 3H), 1.06-0.77 (m, 2H), 0.21-(-0.31) (m, 9H).

Step B: ethyl 5-bromo-2-[(6-{[(22)-3-{[2-(trimethylsilyl)ethoxy]methyl}-2,3-dihydro-1,3benzothiazol-2-ylidene]amino} pyridazin-3-yl)amino]-1,3-thiazol-4-carboxylate.

N-Bromosuccinimide (45.2 mg, 0.25 mmol, 1.3 eq.) was added to a stirred solution of the product from Step A (100 mg, 0.19 mmol, 1 eq.) in dichloromethane (10 ml), and the mixture was stirred at ambient temperature for 1 The solvent was removed in vacuo and the crude product was purified by automated flash column chromatography (CombiFlash Rf, 12 g RediSep™ silica gel cartridge) eluting with a gradient of 0-80% ethyl acetate in isoheptane to give the title product as a cream solid (64.1 mg, 0.11 mmol, 56%) [mixture of isomers].

LC/MS (C ₂₃ H ₂₇ BrN ₆ O ₃ SiS ₂ ) 609 [M+H]+; RT 1.47 (LCMS-V-BI).

1H NMR (400 MHz, DMSOL) δ 12.45 (s, 1H), 12.31 (s, 1H), 8.13 -8.04 (m, 1H), 8.04-7.90 (m, 2H), 7.847.71 (m, 1H), 7.69-7.48 (m, 5H), 7.46-7.32 (m, 3H), 5.98 (s, 2H), 5.92 (s, 2H), 4.41 (qd, J=7.1, 2.4 Hz, 4H), 3.87-3.74 ( m, 4H), 1.43 (td, J=7.1, 1.1 Hz, 6H), 1.03 (s, 2H), 1.08-0.98 (m, 2H), 0.26-(-0.36) (m, 18H).

Synthesis 12: tert-Butyldiphenyl-[2-[[3,5-dimethyl-7-[[5-methyl-4-(4,4,5,5-tetramethyl-1,3,2dioxaborolan-2-yl)pyrazole -1 -yl]methyl]-1 -adamantyl]oxy]ethoxy]silane.

Step A: 3-bromo-5,7-dimethyladamantane-1-carboxylic acid After stirring iron (6.7 g, 120 mmol) in bromine (30.7 ml, 600 mmol, 5 eq.) at 0°C for 1 h, add 3,5-dimethyladamantane1-carboxylic acid (25 g, 1 eq.), and the reaction mixture was stirred at room temperature. within 2 days. After adding EtOAc, the reaction mixture was carefully treated with saturated sodium thiosulfate solution at 0°C and stirred for 15 minutes. After filtering through a pad of Celite and washing with EtOAc, the organic phase was separated, washed with saturated sodium thiosulfate and brine, dried, and concentrated to give the desired product (34.28 g, 74.6%), which was used without further purification.

1H NMR (400 MHz, DMSOL ₆ ): δ ppm 12.33 (br., 1H), 2.21 (s, 2H), 1.96/1.91 (d+d, 4H), 1.50/1.43 (d+d, 4H), 1.21/1.14 (dm+dm, 2H), 0.86 ( s, 6H); ¹³ C NMR (100 MHz, DMSOD6) δ ppm 176.8, 66.8, 54.0, 48.7, 48.5, 45.7, 43.3, 35.5, 29.4; MSVR-ERI (m/z): [M-H] ^- , calculated for C13H ₁₈ BrO ₂ : 285.0496; found: 285.0498.

Step B: 3-bromo-5,7-dimethyl-1-adamantylmethanol.

To the product from step A (34.3 g, 119 mmol) in THF (77.6 ml), a 1 M solution of BH3THF in THF (358 ml, 3 eq.) was slowly added and the reaction mixture was stirred for 18 hours. After adding methanol and stirring within 30 min, purification by column chromatography (silica gel, heptane and MTBE as eluents) gave the desired product (16.19 g, 49.6%).

1H NMR (400 MHz, DMSOO: δ ppm 4.51 (t, 1H), 3.05 (d, 2H), 1.91 (s, 2H), 1.91 (s, 4H), 1.19/1.09 (d+d, 2H ), 1.19/1.05 (d+d, 4H), 0.85 (s, 6H) ¹³ C NMR (100 MHz, DMSOZH) δ ppm 70.4, 68.9, 54.9, 49.8, 49.3, 43.8, 41.4, 35.7, 29.7 ; MSVR-ERI (m/z): [M-Br] ^- , calculated for C ₁₃ H ₂₁ O: 193.1598; found: 193.1589.

Step C: 1-[3-bromo-5,7-dimethyl-1-adamantyl]methyl]pyrazole.

To the product from step B (16.19 g, 59.26 mmol) and 1H-pyrazole (4.841 g, 1.2 eq.) in toluene (178 ml), (cyanomethylene)tributylphosphorane (18.64 ml, 1.2 eq.) was added in one portion and the reaction mixture was stirred at 90°C for 2 hours. Purification by column chromatography (silica gel, heptane and MTBE as eluents) gave the desired product (17.88 g, 93%).

1H NMR (400 MHz, DMSOO: δ ppm 7.63 (d, 1H), 7.43 (d, 1H), 6.23 (t, 1H), 3.90 (s, 2H), 1.92-1.02 (m, 12H), 0.83 (s, 6H); ¹³ C NMR (100 MHz, DMSOO δ ppm 139.0, 131.8, 105.2, 67.7, 61.4, 54.4/48.8/44.6, 50.4, 35.7, 29.6; HRMS-ERI (m/z) : [M] ⁺ , calculated for C ₁₆ H ₂₃ BrN _2: 322.1045; found: 322.1014.

Step D: 5-methyl-1-[[-3-bromo-5,7-dimethyl-1-adamantyl]methyl]pyrazole.

To a solution of the product from step C (17.88 g, 55.3 mmol) in THF (277 ml) at -78°C, butyllithium (2.5 M in THF, 66 ml, 3 eq.) was added, then iodomethane (17.2 ml, 3 eq.) was added after 1 h. 5 eq.). After 10 min, the reaction mixture was quenched with saturated NH ₄ Cl solution, extracted with EtOAc and the combined organic layers were dried and concentrated to give the desired product (18.7 g, 100%), which was used in the next step without further purification.

1H NMR (400 MHz, DMSO<): δ ppm 7.31 (d, 1H), 6.00 (d, 1H), 3.79 (s, 2H), 2.23 (s, 3H), 2.01 (s, 2H), 1.89/1.85 (d+d, 4H), 1.23/1.15 ( d+d, 4H), 1.16/1.05 (d+d, 2H), 0.83 (s, 6H); ¹³ C NMR (100 MHz, DMSOZH) δ ppm 139.2, 138.0, 105.2, 67.8, 57.8, 54.4, 50.6, 48.8, 44.8, 41.5, 35.7, 29.6, 11.8; HRMS-ERI (m/z): [M+H] ⁺ , calculated for C ₁₇ H ₂₆ BrN ₂ : 337.1279; found: 337.1289.

Step E: 2-[[3,5-dimethyl-7-[(5-methylpyrazol-1-yl)methyl]-1-adamantyl]oxy]ethanol.

A mixture of product from step D (18.7 g, 55.3 mmol), ethylene glycol (123 ml, 40 eq.) and DIPEA (48.2 ml, 5 eq.) was stirred at 120°C for 6 hours. The reaction mixture was then diluted with water and extracted with EtOAc, and the combined organic layers were dried and concentrated to give the desired product (18.5 g, 105%), which was used in the next step without further

- 86 046336 cleaning.

¹ H NMR (400 MHz, DMSO-dg): δ ppm 7.29 (d, 1H), 5.99 (d, 1H), 4.45 (t, 1H), 3.78 (s, 2H), 3.39 (q, 2H), 3.32 (t, 2H), 2.23 (s, 3H), 1.34 (s, 2H), 1.27/1.21 (d+d, 4H), 1.13/1.07 (d+d, 4H), 1.04/0.97 (d+d, 2H), 0.84 (s, 6H); ¹³ C NMR (100 MHz, D\1CO-T) δ ppm 139.0, 137.8, 105.1, 74.0, 62.1, 61.5, 58.5, 50.1, 47.0, 46.1, 43.3, 39.7, 33.5, 30.2, 11.9; MSHR-ERI (m/z): [M+H] ⁺ , calculated for C ₁₉ H ₃₁ N ₂ O ₂ : 319.2386; found: 319.2387.

Step F: tert-butyldiphenyl-[2-[[3,5-dimethyl-7-[(5-methylpyrazol-1-yl)methyl]-1adamantyl]oxy]ethoxy]silane.

To a mixture of the product from step E (17.6 g, 55.3 mmol) and imidazole (5.65 g, 1.5 eq.) in DCM (150 ml) was added tert-butylchlorodiphenylsilane (18.6 g, 1.2 eq.) and the reaction mixture was stirred for 1 h Purification by column chromatography (silica gel, heptane and MTBE as eluents) gave the desired product (27.0 g, 87.8%).

1H NMR (400 MHz, DMSO-dA δ ppm 7.72-7.34 (m, 10H), 7.29 (d, 1H), 5.99 (br., 1H), 3.78 (s, 2H), 3.67 (t, 2H ), 3.44 (t, 2H), 2.21 (s, 3H), 1.33 (s, 2H), 1.26/1.18 (d+d, 4H), 1.12/1.06 (d+d, 4H), 1.03/0.96 (d +d, 2H), 0.98 (s, 9H), 0.82 (s, 6H); ¹³ C NMR (100 MHz, DMSOL) δ ppm 139.0, 137.8, 105.1, 74.2, 64.4, 61.7, 58.5, 50.0, 46.9, 46.0, 43.4, 39.6, 33.5, 30.1, 27.1, 19.3, 11.9; HRMS-ERI (m/z): [M+H] ⁺ , calculated for C ₃₅ H ₄₉ N ₂ O ₂ Si: 557.3563; found: 557.3564.

Step G: tert-butyldiphenyl-[2-[[3-[(4-iodo-5-methylpyrazol-1-yl)methyl]-5,7-dimethyl-1adamantyl]oxy]ethoxy]silane.

To a solution of the product from step F (27.0 g, 48.56 mmol) in DMF (243 ml) was added Niodosuccinimide (13.6 g, 1.25 eq.), and the reaction mixture was stirred for 2 hours. After dilution with water, the mixture was extracted with DCM. The combined organic layers were washed with saturated sodium thiosulfate and brine, dried and concentrated to give the desired product (30.1 g, 90%).

1H NMR (400 MHz, DMSO^): δ ppm 7.68-7.37 (m, 10H), 7.45 (s, 1H), 3.89 (s, 2H), 3.67 (t, 2H), 3.44 (t, 2H), 2.23 (s, 3H), 1.30 (s, 2H) , 1.26/1.17 (d+d, 4H), 1.12/1.05 (d+d, 4H), 1.00/0.96 (d+d, 2H), 0.98 (s, 9H), 0.82 (s, 6H); ¹³ C NMR (100 MHz, DMSOO δ ppm 142.5, 140.8, 133.7, 64.4, 61.7, 60.3, 59.9, 49.9, 46.8, 45.9, 43.2, 39.7, 33.5, 30.1, 27.1, 19. 3, 12.2; MSVR-ERI (m/z): [M+H] ⁺ , calculated for C ₃₅ H ₄₈ IN ₂ O ₂ Si: 683.2530; found: 683.2533.

Step H: tert-butyldiphenyl-[2-[[3,5-dimethyl-7-[[5-methyl-4-(4,4,5,5-tetramethyl-1,3,2dioxaborolan-2-yl)pyrazole -1 -yl]methyl]-1 -adamantyl]oxy]ethoxy]silane.

To the product from step G (17.5 g, 25.6 mmol) in THF (128 ml) at 0°C was added chloro(isopropyl)magnesium-Na (1.3 M in THF, 24 ml, 1.2 eq.), stirred for 40 min, treated with 2-isopropoxy-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (15.7 ml, 3 eq.), and the reaction mixture was stirred for 10 min. After dilution with saturated NH4Cl solution and extraction with EtOAc, the combined organic phases were concentrated and purified by column chromatography (silica gel, heptane and MTBE as eluents) to give the expected product (15.2 g, 86.9%).

1H NMR (400 MHz, DMSO^): δ ppm 7.65 (dm, 4H), 7.47 (s, 1H), 7.45 (tm, 2H), 7.40 (tm, 4H), 3.80 (s, 2H), 3.66 (t, 2H), 3.44 (t, 2H), 2.35 (s, 3H), 1.35-0.94 (m, 12H), 1.24 (s, 12H), 0.97 (s, 9H), 0.83 (s, 6H); ¹³ C NMR (100 MHz, DMSO^) δ ppm 146.9, 144.3, 135.6, 130.2, 128.2, 104.7, 83.0, 74.2, 64.4, 61.7, 58.4, 30.1, 27.1, 25.2, 19.3, 12.0; MSHR-ERI (m/z): [M+H] ⁺ , calculated for C ₄₁ H ₆₀ BN ₂ O ₄ Si: 683.4415; found: 683.4423.

Synthesis 13: Methyl 3-bromo-6-(methylamino)pyridine-2-carboxylate.

Step A: Methyl 6-[bis(tert-butoxycarbonyl)amino]-3-bromo-pyridine-2-carboxylate.

Boc2O (59.0 g, 2.5 eq.) was added to methyl 6-amino-3-bromopyridine-2-carboxylate (25.0 g, 108.2 mmol) and DMAP (1.3 g, 0.1 eq.) in DCM (541 ml) at 0°C. , and the reaction mixture was stirred for 2.5 hours. After adding saturated NaHCO ₃ solution and extracting with DCM, the combined organic phases were dried and concentrated to give the expected product (45.0 g, 72.3%).

LC/MS (C ₁₇ H ₂₃ BrN ₂ O ₆ Na) 453 [M+H] ⁺ .

Step B: methyl 3-bromo-6-(tert-butoxycarbonylamino)pyridine-2-carboxylate.

To the product from step A (42.7 g, 74.34 mmol) in DCM (370 ml) at 0°C was added TFA (17.1 ml, 3 eq.), and the reaction mixture was stirred for 18 hours. After washing with saturated NaHCO ₃ solution and brine solution, the combined organic phases were dried, concentrated and purified by column chromatography (silica gel, heptane and EtOAc as eluents) to obtain the target product (28.3 g, 115.2%).

1H NMR (400 MHz, DMSO-dA δ ppm 10.29 (s, 1H), 8.11 (d, 1H), 7.88 (d, 1H), 3.87 (s, 3H), 1.46 (s, 9H) ¹³ C NMR (100 MHz, DMSO^ ₆ ) δ ppm 165.6, 153.1, 151.8/148.3, 143.5, 116.3, 109.2, 53.2, 28.4 LC/MS (C ₁₂ H ₁₅ BrN ₂ O ₄ Na) 353 [M+ H] ⁺ .

Step C: methyl 3-bromo-6-[tert-butoxycarbonyl(methyl)amino]pyridine-2-carboxylate.

To the product from step B (2.96 g, 8.93 mmol) in acetone (45 ml) was added Cs ₂ CO ₃ (8.7 g, 3 eq.) and iodomethane (0.67 ml, 1.2 eq.), and the reaction mixture was stirred for 3 h .After dilution with water

- 87 046336 and extraction with EtOAc, the combined organic phases were washed with brine, dried and concentrated to give the expected product (3.5 g, 112%).

1H NMR (400 MHz, DMSOCH δ ppm 8.13 (d, 1H), 7.78 (d, 1H), 3.90 (s, 3H), 3.27 (s, 3H), 1.47 (s, 9H); ¹³ C NMR (100 MHz, DMSO-dg) δ ppm 165.5, 153.6, 153.6, 147.5, 142.8, 122.5, 111.3, 82.0, 53.3, 34.3, 28.2; MSVR-ERI (m/z): [M+H]+ , calculated for C ₁₃ H ₁₈ BrN ₂ O ₄ : 345.0450; found: 345.0429.

Step D: Methyl 3-bromo-6-(methylamino)pyridine-2-carboxylate.

The product from step C (3.0 g, 8.9 mmol) in 1,1,1,3,3,3-hexafluoroisopropanol (90 ml) was stirred at 100°C for 18 hours. Purification by column chromatography (silica gel, heptane and EtOAc as eluents) led to the obtaining of the target product (2.1 g, 96%).

1H NMR (400 MHz, DMSOO: δ ppm 7.63 (d, 1H), 7.04 (q, 1H), 6.53 (d, 1H), 3.83 (s, 3H), 2.73 (d, 3H); ¹³ C NMR (100 MHz, DMSO<) δ ppm 166.6, 158.2, 148.2, 141.3, 112.1, 101.3, 52.9, 28.3; HRMS-ESI (m/z): [M] ⁺ , calculated for C ₈ H ₉ BrN ₂ O ₂ : 243.9847; found: 243.9843.

Synthesis 14: methyl 3-[1-[[3,5-dimethyl-7-[2-(p-tolylsulfonyloxy)ethoxy]-1-adamantyl]methyl]-5methylpyrazol-4-yl]-6-[methyl-[ 5-methyl-6-[(2)-[3-(2-trimethylsilylethoxymethyl)-1,3-benzothiazol-2ylidene]amino]pyridazin-3-yl]amino]pyridine-2-carboxylate.

Step A: methyl 3-[1-[[3-[2-[tert-butyl(diphenyl)silyl]oxyethoxy]-5,7-dimethyl-1adamantyl]methyl]-5-methylpyrazol-4-yl]-6- (methylamino)pyridine-2-carboxylate.

A mixture of synthesis product 13 (2.07 g, 8.45 mmol), synthesis product 12 (6.9 g, 1.2 eq.), Cs ₂ CO ₃ (8.26 g, 3 eq.) and Pd(AtaPhos) ₂ Cl ₂ (374 mg, 0.1 eq. .) in 1,4-dioxane (51 ml) and water (8.5 ml) were stirred at 80°C for 1 hour. Purification by column chromatography (silica gel, heptane and EtOAc as eluents) gave the desired product (4.5 g, 74%).

1H NMR (400 MHz, DMSO: δ ppm 7.66 (dm, 4H), 7.47-7.38 (m, 6H), 7.31 (d, 1H), 7.23 (s, 1H), 6.78 (q, 1H), 6.59 (d, 1H), 3.82 (s, 2H), 3.67 (t, 2H), 3.58 (s, 3H), 3.46 (t, 2H), 2.77 (d, 3H), 2.06 (s, 3H), 1.35 (s, 2H), 1.27/1.20 (d+d, 4H), 1.14/1.09 (d+d, 4H), 1.05/0.97 (d+d, 2H), 0.98 (s, 9H), 0.84 (s, 6H); ¹³ C NMR (100 MHz, DMSO^) δ ppm 140.1, 137.4, 135.6, 130.2/128.3, 109.8, 74.2, 64.4, 61.7, 58.9, 52.2, 50.0, 46.9, 46.0, 43 .4, 39.8, 33.5, 30.1, 28.4, 27.1, 10.8; HRMS-ERI (m/z): [M+H] ⁺ , calculated for C ₄₃ H ₅₇ N ₄ O ₄ Si: 721.4149; found: 721.4148.

Step B: methyl 3-[1-[[3-[2-[tert-butyl(diphenyl)silyl]oxyethoxy]-5,7-dimethyl-1adamantyl]methyl]-5-methylpyrazol-4-yl]-6- [methyl-[5-methyl-6-[(2)-[3-(2-trimethylsilylethoxymethyl)-1,3benzothiazol-2-ylidene]amino]pyridazin-3-yl]amino]pyridine-2-carboxylate.

Using the general Buchwald reaction procedure II, starting from the product from step A and reacting at reflux for 18 hours, 4.7 g (86%) of the desired product was obtained.

1H NMR (400 MHz, DMSO-dA δ ppm 7.78 (dm, 1H), 7.69-7.36 (m, 10H), 7.63 (q, 1H), 7.63 (d, 1H), 7.47 (dm, 1H) , 7.44 (m, 1H), 7.35 (s, 1H), 7.31 (d, 1H), 7.24 (m, 1H), 5.86 (s, 2H), 3.86 (s, 2H), 3.72 (m, 2H), 3.67 (t, 2H), 3.64 (s, 3H), 3.61 (s, 3H), 3.46 (t, 2H), 2.36 (d, 3H), 2.13 (s, 3H), 1.40-0.94 (m, 12H) , 0.97 (s, 9H), 0.92 (m, 2H), 0.85 (s, 6H), -0.11 (s, 9H); HRMS-ERI (m/z): [M+H] ⁺ , calculated for C ₆₁ H ₇ 9N ₈ O ₅ SSi ₂ : 1091.5433; found: 1091.5426.

Step C: methyl 3-[1-[[3-(2-hydroxyethoxy)-5,7-dimethyl-1-adamantyl]methyl]-5-methylpyrazol-4yl]-6-[methyl-[5-methyl-6 -[(2)-[3-(2-trimethylsilylethoxymethyl)-1,3-benzothiazol-2ylidene]amino]pyridazin-3-yl]amino]pyridine-2-carboxylate.

To the product from step B (1.0 g, 0.916 mmol) in THF (9 ml) at 0°C was added a 1 M solution of TBAF in THF (1.0 ml, 1.1 eq.), and the reaction mixture was stirred for 1 hour. After quenching the reaction mixture with saturated NH4Cl solution and extraction with EtOAc, the combined organic phases were dried, concentrated and purified by column chromatography (silica gel, DCM and MeOH as eluents) to give the expected product (752 mg, 96%).

1H NMR (500 MHz, DMSO^ ₆ ) δ ppm 7.79 (dm, 1H), 7.66 (d, 1H), 7.64 (s, 1H), 7.47 (dm, 1H), 7.43 (m, 1H), 7.36 (s, 1H), 7.33 (d, 1H), 7.25 (m, 1H), 5.87 (s, 2H), 4.46 (t, 1H), 3.86 (s, 2H), 3.73 (m, 2H), 3.68 (s, 3H), 3.62 (s, 3H), 3.40 ( m, 2H), 3.35 (t, 2H), 2.37 (s, 3H), 2.14 (s, 3H), 1.42-0.96 (m, 12H), 0.92 (m, 2H), 0.86 (s, 6H), - 0.10 (s, 9H); MSHR-ERI (m/z): [M+H] ⁺ , calculated for C ₄₅ H ₆₁ N ₈ O ₅ SSi: 853.4255; found: 853.4256.

Step D: methyl 3-[1-[[3,5-dimethyl-7-[2-(p-tolylsulfonyloxy)ethoxy]-1-adamantyl]methyl]-5methylpyrazol-4-yl]-6-[methyl-[ 5-methyl-6-[(2)-[3-(2-trimethylsilylethoxymethyl)-1,3-benzothiazol-2ylidene]amino]pyridazin-3-yl]amino]pyridine-2-carboxylate.

To the product from step C (752 mg, 0.88 mmol) and triethylamine (0.5 ml, 4 eq.) in DCM (4.4 ml) was added p-tolylsulfonyl-4-methylbenzenesulfonate (575.4 mg, 1.76 mmol, 2 eq.), and the reaction the mixture was stirred for 1 hour. Purification by column chromatography (silica gel, heptane and EtOAc as eluents) gave the desired product (722 mg, 81%).

1H NMR (400 MHz, DMSO-dA δ ppm 7.79 (dm, 1H), 7.76 (dm, 2H), 7.68 (d, 1H), 7.64 (s, 1H), 7.47 (m, 1H), 7.46 (dm, 2H), 7.43 (td, 1H), 7.36 (s, 1H), 7.33 (d, 1H), 7.25 (td, 1H), 5.87 (s, 2H), 4.06 (m, 2H), 3.84 ( s, 2H), 3.73 (t, 2H), 3.66 (s, 3H), 3.62 (s, 3H), 3.48 (m, 2H), 2.40 (s, 3H), 2.37 (s, 3H), 2.13 (s , 3H), 1.31-0.94 (m, 12H), 0.92 (t, 2H), 0.83 (s, 6H), -0.10 (s, 9H); ¹³ C NMR (100 MHz, DMSO^ ₆ ) δ ppm .141.2, 137.5, 130.6, 128.1,

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127.2, 123.4, 123.4, 123.1, 114.7, 112.0, 72.9, 71.5, 66.7, 58.8, 58.4, 52.6, 36.6, 30.1, 21.6, 17.8, 17.4, 10.8, 0.9; HRMS-ERI (m/z): [M+H]+, calculated for C ₅₂ H ₆₇ N ₈ O ₇ S ₂ Si: 1007.4343; found: 1007.4344.

Example 1: 2-({6-[(1,3-benzothiazol-2-yl)amino]-4,5-dimethylpyridazin-3-yl}amino)-1,3-thiazol-4carboxylic acid

Step A: ethyl 2-({6-[(1,3-benzothiazol-2-yl)amino]-4,5-dimethylpyridazin-3-yl}amino)-1,3-thiazol4-carboxylate.

Ethyl 2aminothiazole-4-carboxylate (71 mg, 0.41 mmol, 1.2 eq.), synthesis product 9a (100 mg, 0.34 mmol, 1 eq.), xantphos (19.9 mg, 0.03 mmol) were loaded into a microwave-dried vessel for microwave treatment. , 0.1 eq.), tris(dibenzylideneacetone)dipalladium(0) (15.8 mg, 0.02 mmol, 0.05 eq.) and cesium carbonate (134.5 mg, 0.41 mmol, 1.2 eq.). The vessel was evacuated and purged with nitrogen (x3), and then toluene (6 ml) was added. The mixture was bubbled with nitrogen (10 min) and then heated at 180°C for 1.5 h under microwave irradiation. The mixture was partitioned between ethyl acetate (100 ml) and brine (50 ml) and the organic phase was dried (magnesium sulfate) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 12 g RediSep™ silica gel cartridge) eluting with a gradient of 0-100% ethyl acetate in isoheptane gave the desired product (63 mg, 0.15 mmol, 43%).

LC/MS (C ₁₉ H ₁₈ N ₆ O ₂ S ₂ ) 427 [M+H]+; RT 1.32 (LCMS-V-BI).

¹ H NMR (400 MHz, DMSO-06) δ 10.99 (s, 1H), 10.87 (br s, 1H), 7.95 (s, 1H), 7.89 (br s, 1H), 7.57 (br s, 1H), 7.37 (t, J=7.59 Hz, 1H), 7.20 (t, 1H), 4.30 (q, J=7.10 Hz, 2H), 2.38 (s, 3H), 2.36 (s, 3H), 1.32 (t, J =7.11 Hz, 3H).

Step B: 2-({6-[(1,3-benzothiazol-2-yl)amino]-4,5-dimethylpyridazin-3-yl}amino)-1,3-thiazol-4carboxylic acid.

To a solution of the product from step A (38 mg, 0.09 mmol, 1 eq.) in 1,4-dioxane (5 ml) was added a 2M aqueous solution of lithium hydroxide (90 μl, 0.18 mmol, 2 eq.), and the mixture was stirred at temperature environment for 30 minutes, then heated at 40°C for 1 hour, and finally at 80°C for 1 hour. The reaction mixture was then allowed to cool to ambient temperature, the reaction mixture was partitioned between ethyl acetate (10 ml) and water (10 ml) and the pH was adjusted to 6. The aqueous phase was concentrated in vacuo and the residue was triturated with water and collected by filtration. Washing with water, diethyl ether, then dichloromethane, and then drying in vacuo provided the title product as an orange solid (14 mg, 0.04 mmol, 39%).

HRMS-ERI (m/z) [M+H]+, calculated for C ₁₇ H ₁₅ N ₆ O ₂ S ₂ : 399.0692; found: 399.0729.

Example 2: 6-({6-[(1,3-benzothiazol-2-yl)amino]-4,5-dimethylpyridazin-3-yl} amino)pyridin-2-carboxylic acid

Step A: ethyl 6-({6-[(1,3-benzothiazol-2-yl)amino]-4,5-dimethylpyridazin-3-yl}amino)pyridin-2-carboxylate.

Ethyl 6aminopyridine-2-carboxylate (171 mg, 1.03 mmol, 1.5 eq.), synthesis product 9a (200 mg, 0.69 mmol, 1 eq.), xantphos (39.8 mg, 0.07 mmol) were loaded into a microwave-dried vessel for microwave treatment. , 0.1 equiv.), tris(dibenzylideneacetone)diialladium(0) (31.5 mg, 0.03 mmol, 0.05 equiv.) and cesium carbonate (336 mg, 1.03 mmol, 1.5 equiv.). The vessel was evacuated and purged with nitrogen (x3), and then toluene (5 ml) was added. The mixture was bubbled with nitrogen (10 min) and then heated at

- 89 046336

180°C for 3 hours under microwave irradiation. The mixture was diluted with ethyl acetate and water, and filtered through a small pad of celite. The organic layer was washed with brine, dried (magnesium sulfate), filtered and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 24 g RediSep™ silica gel cartridge) eluting with a gradient of 0-100% ethyl acetate in hexanes provided the title compound as a yellow, glassy solid (55 mg, 0.13 mmol, 19 %).

LC/MS (C ₂ iH ₂₀ N ₆ O ₂ S) 421 [M+H]+; RT 1.17 (LCMS-V-BI).

¹ H NMR (400 MHz, DMSO^) δ 10.91 (br s, 1H), 9.17 (s, 1H), 7.85 (br s, 1H), 7.84 (dd, J=7.32, 8.47 Hz, 1H), 7.66 ( d, J=8.47 Hz, 1H), 7.55 (d, J=7.20 Hz, 1H), 7.37 (t, J=7.73 Hz, 1H), 7.19 (s, 1H), 4.32 (q, J=7.13 Hz, 2H), 2.38 (s, 3H), 2.27 (s, 3H), 1.32 (t, J=7.10 Hz, 3H).

Step B: 6-({6-[1,3-benzothiazol-2-yl)amino]-4,5-dimethylpyridazin-3-yl}amino)pyridin-2carboxylic acid.

To a solution of the product from step A (50 mg, 0.12 mmol, 1 eq.) in 1,4-dioxane (5 ml) was added a 2 M aqueous solution of lithium hydroxide (0.12 ml, 0.24 mmol, 2 eq.), and the mixture was heated at 90°C for 1 hour. The mixture was cooled to ambient temperature, the solvent was evaporated in vacuo and the residue was triturated with water and collected by filtration to give the title product as an orange solid (9 mg, 0.02 mmol, 19%).

HRMS-ERI (m/z) [M+H] ⁺ , calculated for C ₁₉ H ₁₇ N ₆ O ₂ S: 393.1128; found: 393.1163.

Example 3: 2-({6-[(1,3-benzothiazol-2-yl)amino]-4,5-dimethylpyridazin-3-yl}amino)-5-(3-{4-[3(dimethylamino) prop-1 -yn-1 -yl]-2-fluorophenoxy}propyl)-1,3-thiazol-4-carboxylic acid

\

Step A: ethyl 2-({6-[(1,3-benzothiazol-2-yl)amino]-4,5-dimethylpyridazin-3-yl}amino)-5-(3-{4-[3(dimethylamino )prop-1 -yn-1 -yl]-2-fluorophenoxy}propyl)-1,3-thiazol-4-carboxylate.

Synthesis product 9a (376 mg, 1.29 mmol, 1.5 eq.), synthesis product 3u (350 mg, 0.86 mmol, 1 eq.), cesium carbonate (394 mg, 1.21 mmol, 1.4 eq.), xanthos (49.9 mg, 0.09 mmol, 0.1 eq.) and tris(dibenzylideneacetone) dipalladium(O) (39.5 mg, 0.04 mmol, 0.05 eq.). The vessel was evacuated and purged with nitrogen (x3), and then trifluorotoluene (10 ml) was added. The mixture was bubbled with nitrogen (10 min) and then heated at 180°C for 1 h under microwave irradiation. Purification by reverse phase automated flash chromatography (CombiFlash Rf, C18 43 g RediSep column) eluting with 5-95% acetonitrile in water gave the title product as a bright yellow glassy solid (266 mg, 0.4 mmol, 47% ).

LC/MS (C33H34FN7O3S2) 66О [M+H] ⁺ ; RT 1.18 (LCMS-V-BI).

1H NMR (4OO MHz, DMSO-d,) δ 11.31 (br s, 1H), 1О.81 (br s, 1H), 7.88 (d, J=7.8 Hz, 1H), 7.5О (d, J=7.8 Hz, 1H), 7.37 (ddd, J=8.2, 7.3, 1.3 Hz, 1H), 7.31 (dd, J=11.9, 2.О Hz, 1H), 7.25-7.11 (m, 3H), 4.26 (q, J=7.1 Hz, 2H), 4.16 (t, J=6.2 Hz, 2H), 3.39 (s, 2H), 3.30 (t, J=7.6 Hz, 2H), 2.37 (s, 3H), 2.35 (s, 3H), 2.21 (s, 6H), 2.14 (q, J=7.2, 6.5 Hz, 2H), 1.29 (t, J=7.1 Hz, 3H).

Step B: 2-({6-[(1,3-benzothiazol-2-yl)amino]-4,5-dimethylpyridazin-3-yl}amino)-5-(3-{4-[3(dimethylamino) prop-1 -yn-1 -yl]-2-fluorophenoxy}propyl)-1,3-thiazol-4-carboxylic acid.

To a solution of the product from step A (266 mg, 0.4 mmol, 1 eq.) in 1,4-dioxane (10 ml) was added a 2 M aqueous solution of lithium hydroxide (0.4 ml, 0.81 mmol, 2 eq. ), and the mixture was stirred at reflux for 3 hours. The reaction mixture was then allowed to cool to ambient temperature, the mixture was concentrated in vacuo and triturated with water (10 ml). The solids were collected by filtration, washed with water and dried in vacuo to give the title product as a bright yellow solid (2O9 mg, 0.33 mmol, 82%) [as lithium salt].

MSVR-ERI (m/z): [M+H] ⁺ , calculated for C ₃₁ H ₃₁ FN ₇ O ₃ S ₂ : 632.1908; found: 632.1914.

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Example 4: 2-({6-[(1,3-benzothiazol-2-yl)amino]pyridazin-3-yl}amino)-1,3-thiazol-4-carboxylic acid

Step A: ethyl 2-({6-[(1,3-benzothiazol-2-yl)amino]pyridazin-3-yl}amino)-1,3-thiazol-4carboxylate.

Synthesis product 9b (500 mg, 1.9 mmol, 1 eq.), ethyl 2-amino-1,3-thiazol-4-carboxylate (393 mg, 2.28 mmol, 1.2 eq.), cesium carbonate was loaded into a flask dried in an oven. (930 mg, 2.85 mmol, 1.5 eq.), xanthos (110 mg, 0.19 mmol, 0.1 eq.) and tris(dibenzylideneacetone) dipalladium(0) (87.1 mg, 0.1 mmol, 0.05 eq.). The flask was evacuated and purged with nitrogen (x3) and then toluene (20 ml) was added. The mixture was then bubbled with nitrogen (15 min) and heated at 180°C for 1.5 h under microwave irradiation. The mixture was partitioned between dichloromethane (200 ml) and water (200 ml) and the organic phase was concentrated in vacuo. The crude material was triturated with dichloromethane and filtered, washed with dichloromethane and dried to give the title product as a yellow solid (417 mg, 1.05 mmol, 55%).

LC/MS (C ₁₇ H ₁₄ N ₆ O ₂ S ₂ ) 397 [M+H]+; RT 1.20 (LCMS-V-BI).

1H NMR (400 MHz, DMSO^) δ 11.86 (s, 2H), 7.97 (s, 1H), 7.95 (s, 1H), 7.65 (d, J=8.1 Hz, 1H), 7.52 (d, J=9.3 Hz, 1H), 7.46-7.32 (m, 2H), 7.23 (t, 1H), 4.29 (q, J=7.1 Hz, 2H), 1.32 (t, J=7.1 Hz, 3H).

Step B: 2-({6-[(1,3-benzothiazol-2-yl)amino]pyridazin-3-yl}amino)-1,3-thiazol-4-carboxylic acid.

To a solution of the product from step A (417 mg, 1.05 mmol, 1 eq.) in 1,4-dioxane (20 ml) was added a 2 M aqueous solution of lithium hydroxide (1.05 ml, 2.09 mmol, 2 eq.), and the reaction mixture was heated reflux for 1 hour. The mixture was allowed to cool to ambient temperature, then the solvent was removed in vacuo and the residue triturated with acetone followed by trituration with a 1:1 isopropanol/water mixture. The solid was collected by filtration and dried in vacuo to give the title product as a brown solid (161 mg, 0.43 mmol, 42%).

HRMS-ERI (m/z) [M+H] ⁺ , calculated for C ₁₅ H _n N ₆ O ₂ S ₂ : 371.0379; found: 371.0404.

Example 5: 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-(propan-2-yl)pyridazin-3-yl}amino)-1,3-thiazol4-carboxylic acid

Step A: ethyl 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-(propan-2-yl)pyridazin-3-yl}amino)-1,3thiazol-4-carboxylate .

Synthesis product 9cb (367 mg, 1.2 mmol, 1 eq.), ethyl 2-amino-1,3-thiazol-4-carboxylate (249 mg, 1.44 mmol, 1.2 eq.) was loaded into an oven-dried vessel for microwave treatment. , cesium carbonate (589 mg, 1.81 mmol, 1.5 eq.), xanthos (69.7 mg, 0.12 mmol, 0.1 eq.) and tris(dibenzylideneacetone) dipalladium(0) (55.1 mg, 0.06 mmol, 0.05 eq.). The vessel was evacuated and purged with nitrogen (x3), and then toluene (10 ml) was added. The mixture was bubbled with nitrogen (15 min) and then heated at 180°C for 1 h under microwave irradiation. The mixture was partitioned between ethyl acetate and water, and the aqueous phase was extracted with ethyl acetate. The combined organics were washed with brine, dried (magnesium sulfate) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 40 g RediSep™ silica gel cartridge) eluting with a gradient of 0-50% ethyl acetate in isoheptane gave the title product as a bright yellow solid (303 mg, 0.69 mmol, 57 %).

LC/MS (C20H20N6O2S2) 441 [M+H] ⁺ ; RT 1.39 (LCMS-V-BI).

1H NMR (400 MHz, DMSO^) δ 12.03 (d, J=233.3 Hz, 1H), 11.72 (s, 1H), 7.94 (s, 1H), 7.85-7.09 (m, 5H), 4.29 (q, J =7.1 Hz, 2H), 3.33 (s, 1H), 1.32 (t, J=7.1 Hz, 3H), 1.25 (d, J=6.8 Hz, 6H).

Step B: 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-(propan-2-yl)pyridazin-3-yl}amino)-1,3-thiazol4-carboxylic acid .

To a solution of the product from step A (300 mg, 0.68 mmol, 1 eq.) in 1,4-dioxane (10 ml) was added 2 M

- 91 046336 aqueous lithium hydroxide solution (0.68 ml, 1.36 mmol, 2 eq.), and the reaction mixture was heated at 60°C for 18 hours. The mixture was allowed to cool to ambient temperature and concentrated in vacuo. The residue was diluted with water and acidified with a 2 M aqueous solution of hydrochloric acid. The resulting solid was collected by filtration, washed with water and dried in vacuo to give a brown solid (130 mg, 0.32 mmol, 46%) [as hydrochloric acid salt].

HRMS-ERI (m/z) [M+H]+, calculated for C ₁₈ H ₁₇ N ₆ O ₂ S ₂ : 413.0849; found: 413.0885.

Example 6: 2-({6-[(1,3-benzothiazol-2-yl)amino]-4-(propan-2-yl)pyridazin-3-yl}amino)-1,3-thiazol4-carboxylic acid

Step A: ethyl 2-({6-[(1,3-benzothiazol-2-yl)amino]-4-(propan-2-yl)pyridazin-3-yl}amino)-1,3thiazol-4-carboxylate .

Synthesis product 9ca (500 mg, 1.64 mmol, 1 eq.), ethyl 2-amino-1,3-thiazol-4-carboxylate (339 mg, 1.97 mmol, 1.2 eq.), xantphos ( 94.9 mg, 0.16 mmol, 0.1 eq.), cesium carbonate (802 mg, 2.46 mmol, 1.5 eq.) and tris(dibenzylideneacetone) dipalladium(0) (75.1 mg, 0.08 mmol, 0.05 eq.). The vessel was evacuated and purged with nitrogen (x3), and then toluene (20 ml) was added. The mixture was bubbled with nitrogen (10 min) and then heated at 200°C for 1.5 h under microwave irradiation. Additional amounts of xanthos (94.9 mg, 0.16 mmol, 0.1 eq.) and tris(dibenzylideneacetone)dipalladium(0) (75.1 mg, 0.08 mmol, 0.05 eq.) were added, the mixture was bubbled with nitrogen (10 min) and then heated at 180°C for 3 hours under microwave irradiation. The reaction mixture was partitioned between ethyl acetate and water, the aqueous phase was extracted with ethyl acetate, and the combined organics were washed with brine, dried (magnesium sulfate) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 40 g RediSep™ silica gel cartridge) eluting with a gradient of 0-55% ethyl acetate in isoheptane provided the title product as an orange solid (147 mg, 0.33 mmol, 20%) .

LC/MS (C20H20N6O2S2) 441 [M+H] ⁺ ; RT 1.35 (LCMS-V-BI).

Step B: 2-({6-[(1,3-benzothiazol-2-yl)amino]-4-(propan-2-yl)pyridazin-3-yl}amino)-1,3-thiazol4-carboxylic acid .

To a solution of the product from step A (147 mg, 0.33 mmol, 1 eq.) in 1,4-dioxane (5 ml) was added a 2 M aqueous solution of lithium hydroxide (0.33 ml, 0.67 mmol, 2 eq.), and the mixture was heated with reflux for 1 hour. The mixture was cooled to ambient temperature and concentrated in vacuo. The residue was added to water and acidified to pH 6. The aqueous phase was extracted with ethyl acetate, and the organics were dried (magnesium sulfate) and concentrated in vacuo. Purification by reverse phase automated flash chromatography (CombiFlash Rf, C18 4.3 g RediSep column) eluting with a gradient of 5-95% acetonitrile in water gave the title product as a yellow solid (7 mg, 0.02 mmol, 5% ).

HRMS-ERI (m/z) [M+H] ⁺ , calculated for C ₁₈ H ₁₇ N ₆ O ₂ S ₂ : 413.0849; found: 413.0886.

Example 7: 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}amino)-1,3-thiazol-4carboxylic acid

Step A: ethyl 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}amino)-1,3-thiazol-4carboxylate.

Synthesis product 9db (400 mg, 1.45 mmol, 1 eq.), ethyl 2-amino-1,3-thiazol-4-carboxylate (373 mg, 2.17 mmol, 1.5 eq.) was loaded into an oven-dried vessel for microwave treatment. , cesium carbonate (706 mg, 2.17 mmol, 1.5 eq.), tris(dibenzylideneacetone)dipalladium(0) (66.2 mg, 0.07 mmol, 0.05 eq.) and xanthos (83.6 mg, 0.14 mmol, 0.1 eq.). The vessel was evacuated and purged with nitrogen (x3), and then toluene (15 ml) was added. The mixture was bubbled with nitrogen (5 min) and then heated at 150°C for 2 h under microwave irradiation. Additional amounts of ethyl 2-amino-1,3 were added

- 92 046336 thiazole-4-carboxylate (375 mg, 2.17 mmol, 1.5 eq.), xanthos (83.6 mg, 0.14 mmol, 0.1 eq.) and tris(dibenzylideneacetone) dipalladium(0) (66.2 mg, 0.07 mmol, 0.05 eq. .), the mixture was bubbled with nitrogen (5 min) and then heated at 180°C for 1 h under microwave irradiation. The mixture was concentrated in vacuo and purified by reverse phase auto flash chromatography (CombiFlash Rf, C18 4.3 g RediSep column) eluting with a gradient of 5-95% acetonitrile in water to give the title product (169 mg, 0.41 mmol, 28%) .

LC/MS (C18H16N6O2S2) 413 [M+H]+; RT 1.27 (LCMS-V-BI).

1H NMR (400 MHz, DMSO-06) δ 11.81 (s, 1H), 11.04 (br s, 1H), 7.94 (s, 1H), 7.90 (br s, 1H), 7.55 (br s, 1H), 7.39 (t, J=7.6 Hz, 1H), 7.21 (t, J=7.5 Hz, 1H), 7.14 (s, 1H), 4.29 (q, J=7.1 Hz, 2H), 2.40 (s, 3H), 1.32 (t, J=7.1 Hz, 3H).

Step B: 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl} amino)-1,3-thiazol-4carboxylic acid.

To a stirred suspension of the product from step A (169 mg, 0.41 mmol, 1 eq.) in 1,4-dioxane (5 ml) was added a 2 M aqueous solution of lithium hydroxide (0.49 ml, 0.49 mmol, 1.2 eq.) and the mixture was stirred at 100°C for 4.5 hours. The reaction mixture was allowed to cool to ambient temperature and concentrated in vacuo. The crude product was suspended in a 2:1 isopropanol/water mixture and stirred for 30 minutes. The solids were collected by filtration and dried in vacuo to give the title product as a bright yellow solid (17 mg, 0.04 mmol, 11%) [as lithium salt].

HRMS-ERI (m/z) [M+H] ⁺ , calculated for C ₁₆ H ₁₃ N ₆ O2S2: 385.0536; found: 385.0567.

Example 8: 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methyl-4-(propan-2-yl)pyridazin-3-yl}amino)-1,3thiazol- 4-carboxylic acid

Step A: ethyl 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methyl-4-(propan-2-yl)pyridazin-3yl}amino)-1,3-thiazole -4-carboxylate.

The synthesis product 9ea (300 mg, 0.94 mmol, 1 eq.), ethyl 2-amino-1,3-thiazol-4-carboxylate (243 mg, 1.41 mmol, 1.5 eq.) was loaded into a microwave-dried vessel for microwave treatment. , cesium carbonate (460 mg, 1.41 mmol, 1.5 eq.), xanthos (218 mg, 0.38 mmol, 0.4 eq.) and tris(dibenzylideneacetone) dipalladium(0) (172.4 mg, 0.18 mmol, 0.2 eq.). The vessel was evacuated and purged with nitrogen (x3), and then toluene (4 ml) was added. The mixture was bubbled with nitrogen (15 min) and then heated at 180°C for 2 h under microwave irradiation. The mixture was concentrated in vacuo and the residue was adsorbed onto Isolute. Purification by reverse phase automated flash chromatography (CombiFlash Rf, C18 4.3 g RediSep column) eluting with a gradient of 5-95% acetonitrile in water gave the target product (22 mg, 0.05 mmol, 5%).

LC/MS (C21H22N6O2S2) 455 [M+H] ⁺ ; RT 1.42 (LCMS-V-BI).

1H NMR (400 MHz, DMSOHO δ 10.96 (s, 1H), 7.92 (s, 1H), 7.87 (br s, 1H), 7.53 (br s, 1H), 7.38 (t, J=7.5 Hz, 1H), 7.20 (t, J=7.6 Hz, 1H), 4.30 (q, J=7.1 Hz, 2H), 3.66 (p, J=7.0 Hz, 1h), 2.48 (s, 3H), 1.35 (d, 6H), 1.32 (t, 3H).

Step B: 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methyl-4-(propan-2-yl)pyridazin-3-yl}amino)-1,3thiazol- 4-carboxylic acid.

To a solution of the product from step A (22 mg, 0.05 mmol, 1 eq.) in tetrahydrofuran (2 ml) and methanol (0.5 ml) was added a 2 M aqueous solution of lithium hydroxide (50 μl, 0.1 mmol, 2 eq.), and the mixture heated at 75°C for 1 hour. The reaction mixture was allowed to cool to ambient temperature and concentrated in vacuo. The resulting solid was transferred back to water (2 ml) and the pH was adjusted to 6. The organics were extracted with ethyl acetate (20 ml), dried (magnesium sulfate), concentrated in vacuo and dried in vacuo to give the title product as a light orange solid substances (19 mg, 0.04 mmol, 92%).

HRMS-ERI (m/z) [M+H] ⁺ , calculated for C ₁₉ H ₁₉ N ₆ O ₂ S2: 427.1005; found: 427.1043.

- 93 046336

Example 9: 2-({6-[(1,3-benzothiazol-2-yl)amino]-4,5-dimethylpyridazin-3-yl}amino)-5-(3phenoxypropyl)-1,3-thiazol-4 -carboxylic acid

Step A: ethyl 2-({6-[(1,3-benzothiazol-2-yl)amino]-4,5-dimethylpyridazin-3-yl}amino)-5-(3phenoxypropyl)-1,3-thiazol- 4-carboxylate.

Synthesis product 9a (45.7 mg, 0.16 mmol, 1 eq.), synthesis product 3v (48.2 mg, 0.16 mmol, 1 eq.), xanthos (9.1 mg, 0.02 mmol, 0.1 eq.) were loaded into a microwave-dried vessel for microwave treatment. .), tris(dibenzylideneacetone)dipalladium(0) (7.2 mg, 0.01 mmol, 0.05 eq.) and cesium carbonate (61.5 mg, 0.19 mmol, 1.2 eq.). The vessel was evacuated and purged with nitrogen (x3), and then toluene (2 ml) was added. The mixture was bubbled with nitrogen (10 min) and then heated at 180°C for 2 h under microwave irradiation. The reaction mixture was partitioned between ethyl acetate and water, then washed successively with water and brine. The organic phase was dried (magnesium sulfate) and the solvent was removed in vacuo. Purification by automated flash column chromatography (Com biFlash Rf, 12 g RediSep™ silica gel cartridge) eluting with a gradient of 0-10% methanol in dichloromethane provided the title product as a brown glass (19.2 mg, 0.03 mmol, 22% ).

LC/MS (C ₂₈ H ₂₈ N ₆ O ₃ S ₂ ) 561 [M+H]+; RT 1.51 (LCMS-V-Bi).

1H NMR (400 MHz, DMSO-de) δ 10.81 (br s, 2H), 7.65 (s, 1H), 7.38 (s, 1H), 7.31 (t, 2H), 7.21 (s, 2H), 7.00-6.89 (m, 3H), 4.27 (q, J=7.2 Hz, 2H), 4.06 (t, J=6.2 Hz, 2H), 3.30 (d, J=8.3 Hz, 2H), 2.37 (s, 3H), 2.35 (s, 3H), 2.13 (q, 2H), 1.30 (t, J=7.1 Hz, 3H).

Step B: 2-({6-[(1,3-benzothiazol-2-yl)amino]-4,5-dimethylpyridazin-3-yl} amino)-5-(3 phenoxypropyl)-1,3-thiazol- 4-carboxylic acid.

To a solution of the product from step A (19 mg, 0.03 mmol, 1 eq.) in 1,4-dioxane (2 ml) was added a 2 M aqueous solution of lithium hydroxide (14.2 mg, 0.34 mmol, 10 eq.), and the mixture was heated with reflux for 10 hours. The reaction mixture was then allowed to cool to ambient temperature, the solvent was removed in vacuo, the residue was triturated with water and the crude product was collected by filtration. Purification by reverse phase preparative HPLC (HPLC-A2 method) provided the title product as a beige solid (3.5 mg, 6.6 µmol, 19%). HRMS-ERI (m/z) [M+H] ⁺ , calculated for C ₂₆ H ₂₅ N ₆ O ₃ S ₂ : 533.1424; found: 533.1460.

Example 10: 2-({6-[(1,3-benzothiazol-2-yl)amino]-4-methyl-5-(propan-2-yl)pyridazin-3-yl}amino)1,3-thiazole -4-carboxylic acid

Step A: ethyl 2-({6-[(1,3-benzothiazol-2-yl)amino]-4-methyl-5-(propan-2-yl)pyridazin-3yl}amino)-1,3-thiazole -4-carboxylate.

Synthesis product 9eb (114 mg, 0.36 mmol, 1 eq.), ethyl 2-amino-1,3-thiazol-4-carboxylate (92.4 mg, 0.54 mmol, 1.5 eq.) was loaded into a microwave-dried vessel for microwave treatment. , cesium carbonate (175 mg, 0.54 mmol, 1.5 eq.), xanthos (41.4 mg, 0.07 mmol, 0.2 eq.) and tris(dibenzylideneacetone) dipalladium(0) (32.7 mg, 0.04 mmol, 0.1 eq.). The vessel was evacuated and purged with nitrogen (x3), and then toluene (4 ml) was added. The mixture was bubbled with nitrogen (15 min) and then heated at 180°C for 1 h under microwave irradiation. The mixture was concentrated in vacuo and purified by reverse phase automated flash chromatography (CombiFlash Rf, C18 4.3 g RediSep column) eluting with a gradient of 5-70% acetonitrile in water to give the title product as a yellow solid (20 mg, 0.04 mmol, 12%).

LC/MS (C ₂₁ H ₂₂ N6O ₂ S ₂ ) 455 [M+H] ⁺ ; RT 1.45 (LCMS-V-Bi).

Step B: 2-({6-[(1,3-benzothiazol-2-yl)amino]-4-methyl-5-(propan-2-yl)pyridazin-3-yl}amino)-1,3thiazol- 4-carboxylic acid.

To a solution of the product from step A (20 mg, 0.04 mmol, 1 eq.) in tetrahydrofuran (2 ml) and methanol

- 94 046336 (0.5 ml) lithium hydroxide monohydrate (3.69 mg, 0.09 mmol, 2 eq.) was added and the reaction mixture was refluxed for 1 hour. The mixture was allowed to cool to ambient temperature and concentrated in vacuo. The product was added to water (3 ml), acidified to pH 6 using

M aqueous hydrochloric acid solution and extracted with a 19:1 mixture of dichloromethane/isopropanol. The organic extract was dried (magnesium sulfate) and concentrated in vacuo to give the title product as a crumbly yellow solid (13.5 mg, 0.03 mmol, 72%).

HRMS-ERI (m/z) [M+H]+, calculated for C ₁₉ H ₁₉ N ₆ O ₂ S ₂ : 427.1005; found: 427.1043.

Example 11: 2-({6-[(1,3-benzothiazol-2-yl)amino]-4-methylpyridazin-3-yl}amino)-1,3-thiazol-4carboxylic acid

Step A: ethyl 2-({6-[(1,3-benzothiazol-2-yl)amino]-4-methylpyridazin-3-yl}amino)-1,3-thiazol-4carboxylate.

Synthesis product 9da (240 mg, 0.87 mmol, 1 eq.), ethyl 2-amino-1,3-thiazol-4-carboxylate (224 mg, 1.3 mmol, 1.5 eq.) was loaded into a microwave-dried vessel for microwave treatment. , cesium carbonate (424 mg, 1.3 mmol, 1.5 eq.), tris(dibenzylideneacetone)dipalladium(0) (39.7 mg, 0.04 mmol, 0.05 eq.) and xanthos (50.2 mg, 0.09 mmol, 0.1 eq.). The vessel was evacuated and purged with nitrogen (x3), and then toluene (15 ml) was added. The mixture was bubbled with nitrogen (5 min) and then heated at 180°C for 3 h under microwave irradiation. The mixture was concentrated in vacuo and purified by reverse phase automated flash chromatography (CombiFlash Rf, C18 26 g RediSep column) eluting with a gradient of 5-95% acetonitrile in water to give the title product as a brown solid (44 mg, 0.11 mmol, 12%).

LC/MS (C ₁₈ H ₁₆ N ₆ O ₂ S ₂ ) 413 [M+H] ⁺ ; RT 1.24 (LCMS-V-BI).

¹ H NMR (400 MHz, DMSOC0 δ 11.61 (br s, 1H), 11.17 (s, 1H), 7.97 (s, 1H), 7.95 (br s, 1H), 7.64 (d, J=8.0 Hz, 1H) , 7.39 (ddd, J=8.2, 7.2, 1.3 Hz, 1H), 7.31 (d, J=1.2 Hz, 1H), 7.22 (ddd, J=8.2, 7.3, 1.2 Hz, 1H), 4.30 (q, J =7.1 Hz, 2H), 2.42 (s, 3H), 1.32 (t, J=7.1 Hz, 3H).

Step B: 2-({6-[(1,3-benzothiazol-2-yl)amino]-4-methylpyridazin-3-yl}amino)-1,3-thiazol-4carboxylic acid.

To a solution of the product from step A (44 mg, 0.11 mmol, 1 eq.) in 1,4-dioxane (5 ml) was added a 2 M aqueous solution of lithium hydroxide (0.21 ml, 0.21 mmol, 2 eq.), and the reaction mixture was heated reflux for 1 hour. The mixture was allowed to cool to ambient temperature, then concentrated in vacuo. The solid was triturated with 4:1 isopropanol/water (4 ml) and stirred for 30 minutes. The solid was collected by filtration, washed with additional cold isopropanol (3 mL) and dried in vacuo to give the title product as a brown solid (33 mg, 0.09 mmol, 80%).

HRMS-ERI (m/z) [M+H] ⁺ , calculated for C ₁₆ H ₁₃ N ₆ O ₂ S ₂ : 385.0536; found: 385.0572.

Example 12: 2-({6-[(1,3-benzothiazol-2-yl)amino]-4,5-dimethylpyridazin-3-yl} amino)-5-(3 methoxypropyl)-1,3-thiazol- 4-carboxylic acid

Step A: ethyl 2-({6-[(1,3-benzothiazol-2-yl)amino]-4,5-dimethylpyridazin-3-yl}amino)-5-(3methoxypropyl)-1,3-thiazol- 4-carboxylate.

Synthesis product 9a (260 mg, 0.89 mmol, 1 eq.), synthesis product 3w (219 mg, 0.89 mmol, 1 eq.), xantphos (51.8 mg, 0.09 mmol, 0.1 eq.) were loaded into a microwave-dried vessel for microwave treatment. .), tris(dibenzylideneacetone)dipalladium(0) (41 mg, 0.04 mmol, 0.05 eq.) and cesium carbonate (350 mg, 1.07 mmol, 1.2 eq.). The vessel was evacuated and purged with nitrogen (x3), and then toluene (5 ml) was added. The mixture was bubbled with nitrogen (10 min) and then heated at 180°C for 1 h under microwave irradiation. The reaction mixture was diluted with ethyl acetate, washed successively with water and brine, then dried (magnesium sulfate) and the solvent was removed in vacuo. Purification by reverse phase automated flash chromatography (CombiFlash Rf, C18 13 g column Re

- 95 046336 diSep) eluting with 5-95% acetonitrile in water gave the target product as a yellow glassy substance (72.5 mg, 0.15 mmol, 16%).

LC/MS (C ₂₃ H ₂₆ N ₆ O ₃ S ₂ ) 499 [M+H]+; RT 1.38 (LCMS-V-BI).

¹ H NMR (400 MHz, DMSO^) δ 10.76 (br s, 1H), 7.88 (d, J=7.7 Hz, 1H), 7.50 (br s, 1H), 7.37 (td, J=8.1, 7.7, 1.3 Hz, 1H), 7.20 (t, J=7.7 Hz, 1H), 4.29 (q, J=7.1 Hz, 2H), 3.41 (t, J=6.3 Hz, 1H), 3.27 (s, 3H), 3.21- 3.12 (m, 2H), 2.37 (s, 3H), 2.35 (s, 3H), 1.96-1.84 (m, 2H), 1.32 (t, J=7.1 Hz, 3H).

Step B: 2-({6-[(1,3-benzothiazol-2-yl)amino]-4,5-dimethylpyridazin-3-yl}amino)-5-(3methoxypropyl)-1,3-thiazol-4 -carboxylic acid.

To a solution of the product from step A (72.5 mg, 0.15 mmol, 1 eq.) in 1,4-dioxane (5 ml) was added lithium hydroxide monohydrate (12.2 mg, 0.29 mmol, 2 eq.), and the mixture was refluxed at for 10 hours. The reaction mixture was allowed to cool to ambient temperature and the solvent was removed in vacuo. The resulting residue was triturated with water, then collected by filtration, washed with water and dried in vacuo to give the title product as a brown solid (40.9 mg, 0.09 mmol, 60%) [as lithium salt].

HRMS-ERI (m/z) [M+H] ⁺ , calculated for C ₂₁ H ₂₃ N ₆ O ₃ S ₂ : 471.1268; found: 471.1306.

Example 13: 2-({6-[(1,3-benzothiazol-2-yl)amino]-4,5-dimethylpyridazin-3-yl}amino)-5-[3-(2fluorophenoxy)propyl]-1, 3-thiazole-4-carboxylic acid

Step A: ethyl 2-({6-[(1,3-benzothiazol-2-yl)amino]-4,5-dimethylpyridazin-3-yl}amino)-5-[3-(2fluorophenoxy)propyl]-1 ,3-thiazole-4-carboxylate.

Synthesis product 9a (162 mg, 0.56 mmol, 1 eq.), synthesis product 3x (181 mg, 0.56 mmol, 1 eq.), xantphos (36.0 mg, 0.06 mmol, 0.11 eq.) were loaded into a microwave-dried vessel for microwave treatment. .), tris(dibenzylideneacetone)dipalladium(0) (25.6 mg, 0.03 mmol, 0.05 eq.) and cesium carbonate (243 mg, 0.75 mmol, 1.34 eq.). The vessel was evacuated and purged with nitrogen (x3), and then toluene (4 ml) and dimethylformamide (1 ml) were added. The mixture was bubbled with nitrogen (10 min) and then heated at 180°C for 1 h under microwave irradiation. The reaction mixture was diluted with ethyl acetate, washed successively with water and brine, dried (magnesium sulfate) and concentrated in vacuo. Purification by reverse phase automated flash chromatography (CombiFlash Rf, C18 13 g RediSep column) eluting with a gradient of 5-95% acetonitrile in water gave the title product as a brown glassy solid (76.1 mg, 0.13 mmol, 24% ).

LC/MS (C ₂₈ H ₂ 7FN ₆ O ₃ S ₂ ) 579 [M+H] ⁺ ; RT 1.49 (LCMS-V-BI).

¹ H NMR (400 MHz, DMSO^) δ 10.78 (br s, 1H), 7.89 (brs, 1H), 7.42-7.28 (m, 2H), 7.28-7.08 (m, 4H), 7.00-6.86 (m, 1H), 4.26 (q, J=7.1 Hz, 2H), 4.14 (t, J=6.2 Hz, 2H), 3.30 (d, J=6.8 Hz, 2H), 2.37 (s, 3H), 2.35 (s, 3H), 2.21-2.11 (m, 2H), 1.30 (t, J=7.1 Hz, 3H).

Step B: 2-({6-[(1,3-benzothiazol-2-yl)amino]-4,5-dimethylpyridazin-3-yl}amino)-5-[3-(2fluorophenoxy)propyl]-1, 3-thiazole-4-carboxylic acid.

To a solution of the product from step A (76 mg, 0.13 mmol, 1 eq.) in 1,4-dioxane (5 ml) was added lithium hydroxide monohydrate (11.02 mg, 0.26 mmol, 2 eq.), and the mixture was refluxed at for 8 hours. The mixture was allowed to cool to ambient temperature and the solvent was removed in vacuo. Purification by reverse phase automated flash chromatography (CombiFlash Rf, C18 13 g RediSep column) eluting with a gradient of 5-95% acetonitrile in water yielded a substance which was further purified by reverse phase preparative HPLC (HPLC method' -A2) to obtain the target product as a beige solid (7.2 mg, 0.01 mmol, 10%).

MSHR-ERI (m/z) [M+H] ⁺ , calculated for C ₂₆ H ₂₄ FN ₆ O ₃ S ₂ : 551.1330; found: 551.1368.

- 96 046336

Example 14: 2-({6-[(1,3-benzothiazol-2-yl)amino]-4,5-dimethylpyridazin-3-yl} amino)-5-{3-[4(dimethylcarbamoyl)-2- fluorophenoxy]propyl}-1,3-thiazole-4-carboxylic acid

Step A: ethyl 2-({6-[(1,3-benzothiazol-2-yl)amino]-4,5-dimethylpyridazin-3-yl}amino)-5-{3-[4(dimethylcarbamoyl)-2 -fluorophenoxy]propyl}-1,3-thiazole-4-carboxylate.

Synthesis product 9a (143 mg, 0.49 mmol, 1 eq.), synthesis product 3y (199 mg, 0.49 mmol, 1 eq.), xanthos (28.5 mg, 0.05 mmol, 0.1 eq.) were loaded into a microwave-dried vessel for microwave treatment. .), tris(dibenzylideneacetone)dipalladium(0) (22.5 mg, 0.02 mmol, 0.05 eq.) and cesium carbonate (192 mg, 0.59 mmol, 1.2 eq.). The vessel was evacuated and purged with nitrogen (x3), and then toluene (4 ml) and dimethylformamide (1 ml) were added. The mixture was bubbled with nitrogen (10 min) and then heated at 180°C for 1 h under microwave irradiation. The reaction mixture was diluted with ethyl acetate, then washed with water, followed by brine. The organic extract was dried (magnesium sulfate) and the solvent was removed in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 12 g RediSep™ silica gel cartridge) eluting with a gradient of 0-10% methanol in dichloromethane gave the title product as a brown glass (60 mg, 0.09 mmol, 19%) .

LC/MS (C ₃₁ H ₃₂ FN ₇ O ₄ S ₂ ) 650 [M+H]+; RT 1.36 (LCMS-V-BI).

1H NMR (400 MHz, DMSO-06) δ 10.77 (br s, 1H), 7.87 (s, 1H), 7.42-7.28 (m, 2H), 7.26-7.17 (m, 4H), 4.26 (q, J= 7.1 Hz, 2H), 4.19 (t, J=6.2 Hz, 2H), 2.97-2.94 (m, 2H), 2.93 (s, 6H), 2.37 (s, 3H), 2.35 (s, 3H), 2.18 ( q, J=7.3, 6.9 Hz, 2H), 1.29 (t, 3H).

Step B: 2-({6-[(1,3-benzothiazol-2-yl)amino]-4,5-dimethylpyridazin-3-yl}amino)-5-{3-[4(dimethylcarbamoyl)-2- fluorophenoxy]propyl}-1,3-thiazole-4-carboxylic acid.

To a solution of the product from step A (60 mg, 0.09 mmol, 1 eq.) in 1,4-dioxane was added lithium hydroxide monohydrate (19.4 mg, 0.46 mmol, 5 eq.) and the mixture was refluxed for 14 h. The reaction mixture was allowed to cool to ambient temperature and the solvent was removed in vacuo. The residue was triturated with water followed by trituration with ethanol, and then purified by reverse phase preparative HPLC (HPLC-U-A2 method) to give the title product as a brown solid (11.6 mg, 0.02 mmol, 20%).

MSHR-ERI (m/z) [M+H] ⁺ , calculated for C ₂₉ H ₂₉ FN ₇ O ₄ S ₂ : 622.1701; found: 622.1740.

Example 15: 2-({6-[(1,3-benzothiazol-2-yl)amino]-4-ethyl-5-methylpyridazin-3-yl}amino)-1,3-thiazol4-carboxylic acid

Step A: ethyl 2-({6-[(1,3-benzothiazol-2-yl)amino]-4-ethyl-5-methylpyridazin-3-yl}amino)-1,3thiazol-4-carboxylate.

The synthesis product 9fa (400 mg, 1.31 mmol, 1 eq.), ethyl 2-amino-1,3-thiazol-4-carboxylate (316 mg, 1.84 mmol, 1.4 eq.) was loaded into a microwave-dried vessel for microwave treatment. , cesium carbonate (599 mg, 1.84 mmol, 1.4 eq.), xanthos (75.9 mg, 0.13 mmol, 0.1 eq.) and tris(dibenzylideneacetone)diialladium(0) (60.1 mg, 0.07 mmol, 0.05 eq.). The vessel was evacuated and purged with nitrogen (x3), and then toluene (15 ml) was added. The mixture was bubbled with nitrogen (10 min) and then heated at 200°C for 2 h under microwave irradiation. The reaction mixture was concentrated in vacuo and purified by reverse phase automatic flash chromatography (CombiFlash Rf, C18 13 g RediSep column) eluting with a gradient of 5-95% acetonitrile in water to give the target product (70 mg, 0.16 mmol, 12% ).

LC/MS (C20H20N6O2S2) 441 [M+H] ⁺ ; RT 1.37 (LCMS-V-BI).

1H NMR (400 MHz, DMSO^6) δ 11.08 (s, 1H), 10.88 (br s, 1H), 7.94 (s, 1H), 7.89 (br s, 1H), 7.53 (br s,

- 97 046336

1H), 7.38 (t, 1H), 7.20 (t, 1H), 4.30 (q, J=7.1 Hz, 2H), 2.93 (q, J=7.4 Hz, 2H), 2.40 (s, 3H), 1.32 ( t, J=7.1 Hz, 3h), 1.09 (t, J=7.4 Hz, 3H).

Step B: 2-({6-[(1,3-benzothiazol-2-yl)amino]-4-ethyl-5-methylpyridazin-3-yl} amino)-1,3-thiazol-4carboxylic acid.

To a solution of the product from step A (70 mg, 0.16 mmol, 1 eq.) in 1,4-dioxane (4 ml) was added lithium hydroxide monohydrate (0.32 ml, 0.32 mmol, 2 eq.) and the mixture was refluxed at for 1 hour. The reaction mixture was allowed to cool to ambient temperature and concentrated in vacuo. The residue was then suspended in water, the pH was adjusted to 6, and the mixture was then reconcentrated. The solid was triturated with isopropanol with a few drops of water, then collected by filtration, washed with isopropanol and dried in vacuo to give the title product as a brown solid (38.8 mg, 0.09 mmol, 59%).

MSVR-ERI (m/z) [MH] ^- , calculated for C ₁₈ H ₁₅ N ₆ O ₂ S ₂ : 411.0703; found: 411.0707.

Example 4-carbon

Stage

16: 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-ethyl-4-methylpyridazin-3-yl}amino)-1,3-thiazole acid

A: ethyl 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-ethyl-4-methylpyridazin-3-yl}amino)-1,3 thiazol-4-carboxylate.

Synthesis product 9fb (300 mg, 0.98 mmol, 1 eq.), ethyl 2-amino-1,3-thiazol-4-carboxylate (220 mg, 1.28 mmol, 1.3 eq.) was loaded into a microwave-dried vessel for microwave treatment. , cesium carbonate (449 mg, 1.38 mmol, 1.4 eq.), xanthos (57 mg, 0.1 mmol, 0.1 eq.) and tris(dibenzylideneacetone) dipalladium(0) (45.1 mg, 0.05 mmol, 0.05 eq.). The vessel was evacuated and purged with nitrogen (x3), and then toluene (15 ml) was added. The mixture was bubbled with nitrogen (10 min) and then heated at 200°C for 2 h under microwave irradiation. The reaction mixture was concentrated in vacuo and purified by reverse phase automatic flash chromatography (CombiFlash Rf, C18 13 g RediSep column) eluting with 5-95% acetonitrile in water to give the title product as a yellow solid (54 mg, 0.12 mmol , 12%).

LC/MS (C ₂₀ H ₂₀ N ₆ O ₂ S ₂ ) 441 [M+H]+; RT 1.38 (LCMS-V-Bi).

1H NMR (400 MHz, DMSO-06) δ 10.98 (s, 1H), 7.95 (s, 1H), 7.84 (s, 1H), 7.43 (br s, 1H), 7.36 (t, J=7.5 Hz, 1H ), 7.18 (t, J=7.5 Hz, 1H), 4.30 (q, J=7.1 Hz, 2H), 2.92 (q, J=7.5, 7.1 Hz, 2H), 2.38 (s, 3H), 1.32 (t , J=7.1 Hz, 3H), 1.11 (t, J=7.4 Hz, 3H).

Step B: 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-ethyl-4-methylpyridazin-3-yl}amino)-1,3-thiazol-4carboxylic acid.

To a solution of the product from step A (54 mg, 0.12 mmol, 1 eq.) in 1,4-dioxane (4 ml) was added a 2 M aqueous solution of lithium hydroxide (0.12 ml, 0.25 mmol, 2 eq.), and the reaction mixture was heated reflux for 2 hours. The mixture was concentrated in vacuo, then suspended in water and the pH was adjusted to 5-6, after which the mixture was concentrated in vacuo again. The crude solid was triturated with water, stirred for 30 min, then collected by filtration, washed with cold water and dried in vacuo to give the title product as a yellow solid (28.9 mg, 0.07 mmol, 57%).

HRMS-ERI (m/z) [M+H]+, calculated for C ₁₈ H ₁₇ N ₆ O ₂ S ₂ : 413.0849; found: 413.0848.

Example 17: 2-[[4-(1,3-Benzothiazol-2-ylamino)-5,6,7,8-tetrahydrophthalazin-1-yl]amino]thiazol-4carboxylic acid

Stage A: 4-bromo-5,6,7,8-tetrahydrophthalazine-1-amine.

A mixture of 4.0 g of 1,4-dibromo-5,6,7,8-tetrahydrophthalazine (13.7 mmol) in 50 ml of a 25% solution of NH ₃ in water was kept at 100°C for 18 hours. After concentration, the residue was added to methanol and treated charcoal, stirred, filtered, concentrated, added to EtOAc, washed

- 98 046336

10% NaOH solution, dried over MgSO ₄ , filtered and concentrated to obtain 1.28 g (41%) of the target product.

LC-MS-ESI (m/z): [M+H]+, calculated for C ₈ H ₁₁ BrN ₃ : 228, 230; found: 228, 230.

Step B: ^(4-bromo-5,6,7,8-tetrahydrophthalazin-1-yl)-1,3-benzothiazol-2-amine.

A mixture of 1.05 g of the product from step A (4.6 mmol, 1 eq.), 1.8 g of 2-bromo-1,3-benzothiazole (8.4 mmol, 1.8 eq.) and 736 mg of sodium hydride (60% suspension, 18.4 mmol, 3.6 eq. .) in 80 ml of 1,4-dioxane was kept under reflux for 0.5 h. After cooling and quenching the reaction mixture with methanol, the mixture was concentrated, treated with water, and the product was filtered and dried. The solid was triturated with EtOAc, filtered and dried to give 1.56 g (94%) of the expected product.

1H NMR (500 MHz, DMSO-de) δ ppm 11.51 (br., 1H), 7.83 (d, 1H), 7.53 (brd., 1H), 7.38 (td, 1H), 7.22 (td, 1H), 2.75/2.62 (m+m, 4H), 1.79 ( m, 4H); ¹³ C NMR (125 MHz, DMSO^) δ ppm 126.6, 122.9, 122.2, 28.8/24, 21.4/21.1; MSHR-ERI (m/z): [M+H]+, calculated for C ₁₅ H ₁₄ BrN ₄ S: 361.0117; found: 361.0107.

Step C: ethyl 2-[[4-(1,3-benzothiazol-2-ylamino)-5,6,7,8-tetrahydrophthalazin-1-yl]amino]thiazol-4carboxylate.

A mixture of 120 mg of the product from step B (0.33 mmol, 1 eq.), 57 mg of ethyl 2-aminothiazole-4-carboxylate (0.33 mmol, 1 eq.), 12 mg of Pd ₂ (dba) ₃ (0.013 mmol, 0.04 eq. ), 19 mg of xanthos (0.033 mmol, 0.1 eq.) and 217 mg of Cs ₂ CO ₃ (0.66 mmol, 2 eq.) in 10 ml of trifluoromethylbenzene were kept at 200°C for 0.5 h. The reaction mixture was concentrated and purified using a preparative HPLC to obtain 31 mg (21%) of the desired product.

1H NMR (500 MHz, DMSO-d^ δ ppm 7.90 (s, 1H), 7.86 (br., 1H), 7.55 (br., 1H), 7.36 (t, 1H), 7.19 (t, 1H ), 4.31 (q, 2H), 2.75 (br., 4H), 1.81 (br., 4H), 1.33 (t, 3H); ¹³ C NMR (125 MHz DMSO-de) δ ppm 126.3, 122.5 , 121.5; MSHR-ERI (m/z): [M+H]+, calculated for C ₂₁ H ₂₁ N ₆ O ₂ S ₂ : 453.1162; found: 453.1151.

Step D: 2-[[4-(1,3-benzothiazol-2-ylamino)-5,6,7,8-tetrahydrophthalazin-1-yl]amino]thiazol-4carboxylic acid.

A mixture of 31 mg of the product from step C (0.069 mmol, 1 eq.), 5.8 mg of LiOHxH ₂ O (2 eq.) and 8 drops of water in 10 ml of 1,4-dioxane was kept under reflux for 2 hours. the mixture was concentrated and purified by preparative HPLC to obtain 24 mg (83%) of the desired product.

MSHR-ERI (m/z): [M+H]+, calculated for C ₁₉ H ₁₇ N ₆ O ₂ S ₂ : 425.0849; found: 425.0854.

Example 18: 2-({6-[(1,3-benzothiazol-2-yl)amino]-4-(hydroxymethyl)-5-methylpyridazin-3-yl}amino)1,3-thiazol-4-carboxylic acid

Step A: ethyl 2-({6-[(1,3-benzothiazol-2-yl)amino]-4-[(benzyloxy)methyl]-5-methylpyridin-3yl} amino)-1,3-thiazol-4 -carboxylate.

Synthesis product 9ga (176 mg, 0.44 mmol, 1 eq.), ethyl 2-amino-1,3-thiazol-4-carboxylate (115 mg, 0.67 mmol, 1.5 eq.) was loaded into a microwave-dried vessel for microwave treatment. , cesium carbonate (202 mg, 0.62 mmol, 1.4 eq.), tris(dibenzylideneacetone)dipalladium(0) (40.6 mg, 0.04 mmol, 0.1 eq.) and xanthos (12.8 mg, 0.02 mmol, 0.05 eq.). The vessel was evacuated and purged with nitrogen (x3), and then toluene (15 ml) was added. The mixture was bubbled with nitrogen (5 min) and then heated at 180°C for 1 h under microwave irradiation. The mixture was concentrated in vacuo and purified by reverse phase auto flash chromatography (CombiFlash Rf, C18 24 g RediSep column) eluting with a gradient of 5-95% acetonitrile in water to give the title product (82 mg, 0.15 mmol, 35%) .

LC/MS (C ₂₆ H ₂₄ N ₆ O ₃ S ₂ ) 533 [M+H]+; RT 1.44 (LCMS-V-BI).

1H NMR (400 MHz, DMSO-d^δ 10.91 (br s, 1H), 7.96 (s, 1H), 7.88 (br s, 1H), 7.56 (br s, 1H), 7.43-7.26 (m, 6H) , 7.23-7.18 (m, 1H), 4.87 (s, 2H), 4.65 (s, 2H), 4.31 (q, J=7.1 Hz, 2H), 2.40 (s, 3H), 1.33 (t, J=7.1 Hz, 3H).

Step B: ethyl 2-({6-[(1,3-benzothiazol-2-yl)amino]-4-(hydroxymethyl)-5-methylpyridin-3yl} amino)-1,3-thiazol-4-carboxylate.

A solution of the product from step A (72 mg, 0.14 mmol, 1 eq.) in dichloromethane (5 ml) was cooled to 0°C and a 1 M solution of boron trichloride (0.68 ml, 0.68 mmol, 5 eq.) was added dropwise. The mixture was then allowed to warm to ambient temperature over 2 hours. The reaction mixture was quenched by adding methanol and concentrated in vacuo. The resulting solid was triturated with dichlor

- 99 046336 methane, filtered, washed with dichloromethane and dried in vacuo to give the title product as a cream solid (54 mg, 0.12 mmol, 90%).

LC/MS (C19H18N6O3S2) 443 [M+H]+; RT 1.23 (LCMS-V-BI).

¹ H NMR (400 MHz, DMSO^) δ 7.97 (s, 1H), 7.89 (d, J=7.7 Hz, 1H), 7.51 (d, J=8.0 Hz, 1H), 7.39 (ddd, J=8.2, 7.3, 1.3 Hz, 1H), 7.22 (td, J=7.5, 1.2 Hz, 1H), 4.83 (s, 2H), 4.30 (q, J=7.1 Hz, 2H), 2.43 (s, 3H), 1.32 ( t, J=7.1 Hz, 3H).

Step C: 2-({6-[(1,3-benzothiazol-2-yl)amino]-4-(hydroxymethyl)-5-methylpyridin-3-yl}amino)1,3-thiazol-4-carboxylic acid .

To a solution of the product from step B (50 mg, 0.11 mmol, 1 eq.) in 1,4-dioxane (5 ml) was added a 2 M aqueous solution of lithium hydroxide (0.56 ml, 0.56 mmol, 5 eq.), and the mixture was heated with reflux for 30 minutes. The mixture was cooled to ambient temperature and concentrated in vacuo. The solid was added to water (5 ml) and neutralized to pH 6 with 1 M HCl, and then concentrated in vacuo. The solid was then triturated with 4:1 isopropanol/water and stirred for 30 minutes. The solids were collected by filtration and dried in vacuo to give the title product as an orange solid (30 mg, 0.07 mmol, 64%).

HRMS-ERI (m/z) [M+H]+, calculated for C ₁₇ H ₁₅ N ₆ O3S ₂ : 415.0642; found: 415.0638.

Example 19: 2-({6-[(1,3-benzothiazol-2-yl)amino]-4-cyclopropyl-5-methylpyridazin-3-yl}amino)1,3-thiazol-4-carboxylic acid

Step A: ethyl 2-({6-[(1,3-benzothiazol-2-yl)amino]-4-cyclopropyl-5-methylpyridazin-3yl}amino)-1,3-thiazol-4-carboxylate.

Synthesis product 9ha (300 mg, 0.95 mmol, 1 eq.), ethyl 2-amino-1,3-thiazol-4-carboxylate (245 mg, 1.42 mmol, 1.5 eq.) was loaded into a microwave-dried vessel for microwave treatment. , cesium carbonate (463 mg, 1.42 mmol, 1.5 eq.), xanthos (54.8 mg, 0.09 mmol, 0.1 eq.) and tris(dibenzylideneacetone) dipalladium(0) (43.4 mg, 0.05 mmol, 0.05 eq.). The vessel was evacuated and purged with nitrogen (x3), and then alpha,alpha,alpha-trifluorotoluene (10 ml) was added. The mixture was bubbled with nitrogen (5 min) and then heated at 180°C for 1 h under microwave irradiation. The reaction mixture was concentrated in vacuo and purified by reverse phase automated flash chromatography (CombiFlash Rf, C18 26 g RediSep column) eluting with a gradient of 5-95% acetonitrile (with 0.08% formic acid as modifier) in water (with 0.08% formic acid). acid as a modifier) to obtain the target product as a bright yellow solid (67 mg, 0.15 mmol, 16%).

LC/MS (C ₂₁ H ₂₀ N ₆ 0 ₂ S ₂ ) 453 [M+H]+; RT 1.34 (LCMS-V-BI).

1H NMR (400 MHz, DMSO-4 ₆ ) δ 11.37 (br s, 1H), 10.56 (s, 1H), 7.96 (s, 1H), 7.89 (d, J=7.7 Hz, 1H), 7.51 (s, 1H), 7.38 (td, J=7.7, 1.3 Hz, 1H), 7.20 (t, 1H), 4.30 (q, J=7.1 Hz, 2H), 2.49 (s, 3H), 1.90 (q, 1H), 1.31 (t, 3H), 1.27-1.21 (m, 2H), 0.59 (dt, J=5.9, 3.1 Hz, 2H).

Step B: 2-({6-[(1,3-benzothiazol-2-yl)amino]-4-cyclopropyl-5-methylpyridazin-3-yl}amino)-1,3thiazol-4-carboxylic acid.

To a solution of the product from step A (67 mg, 0.15 mmol, 1 eq.) in 1,4-dioxane (5 ml) was added a 2 M aqueous solution of lithium hydroxide (0.15 ml, 0.3 mmol, 2 eq.), and the mixture was heated with reflux for 1 hour. The reaction mixture was concentrated in vacuo, then added to water and the pH was adjusted to 6 with 1N. an aqueous solution of hydrochloric acid, and then re-concentrated in vacuo. Trituration with 5:1 isopropanol/water (5 mL) resulted in a solid, which was collected by filtration, washed with isopropanol and dried in vacuo to give the title product as a yellow solid (28 mg, 0.07 mmol, 45%).

HRMS-ERI (m/z) [M+H]+, calculated for C ₁₉ H ₁₇ N ₆ O ₂ S ₂ : 425.0849; found: 425.0857.

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Example 20: 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-cyclopropyl-4-methylpyridazin-3-yl}amino)1,3-thiazol-4-carboxylic acid

Step A: ethyl 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-cyclopropyl-4-methylpyridazin-3yl}amino)-1,3-thiazol-4-carboxylate.

Synthesis product 9hb (219 mg, 0.69 mmol, 1 eq.), ethyl 2-amino-1,3-thiazol-4-carboxylate (179 mg, 1.04 mmol, 1.5 eq.) was loaded into a microwave-dried vessel for microwave treatment. , cesium carbonate (338 mg, 1.04 mmol, 1.5 eq.), xanthos (40 mg, 0.07 mmol, 0.1 eq.) and tris(dibenzylideneacetone) dipalladium(0) (31.7 mg, 0.03 mmol, 0.05 eq.). The vessel was evacuated and purged with nitrogen (x3) and then alpha,alpha,alpha-trifluorotoluene (15 ml) was added. The mixture was bubbled with nitrogen (5 min) and then heated at 170°C for 1 h under microwave irradiation. The reaction mixture was concentrated in vacuo and purified by reverse phase automated flash chromatography (CombiFlash Rf, C18 24 g RediSep column) eluting with a gradient of 5-95% acetonitrile (containing 0.08% formic acid) in water (containing 0.08% formic acid) with obtaining the target product (54 mg, 0.12 mmol, 17%).

LC/MS (C ₂₁ H ₂₀ N ₆ O ₂ S ₂ ) 453 [M+H] ⁺ ; RT 1.16 (LCMS-V-BI).

1H NMR (400 MHz, DMSO-de) δ 10.86 (br s, 1H), 10.56 (s, 1H), 7.96 (s, 1H), 7.87 (s, 1H), 7.55 (br s, 1H), 7.38 ( t, J=7.5 Hz, 1H), 7.20 (t, J=7.5 Hz, 1H), 4.30 (q, J=7.1 Hz, 2H), 2.49 (s, 3h), 1.91 (p, J=7.7 Hz, 1H), 1.33 (t, 3H), 1.27-1.21 (m, 2H), 0.59 (dt, J=5.9, 3.1 Hz, 2H).

Step B: 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-cyclopropyl-4-methylpyridazin-3-yl}amino)-1,3thiazol-4-carboxylic acid.

To a solution of the product from step A (54 mg, 0.12 mmol, 1 eq.) in 1,4-dioxane (5 ml) was added a 2 M aqueous solution of lithium hydroxide (0.24 ml, 0.24 mmol, 2 eq.), and the mixture was heated with reflux for 1 hour. The reaction mixture was allowed to cool to ambient temperature and concentrated in vacuo. The residue was taken up in water and neutralized to pH 6 with 1 M aqueous hydrochloric acid and then concentrated in vacuo. The solid was triturated with 4:1 isopropanol/water and stirred for 30 minutes. The solids were collected by filtration, washed with cold isopropanol and dried in vacuo to give the title product as an orange solid (30 mg, 0.07 mmol, 59%).

HRMS-ERI (m/z) [M+H] ⁺ , calculated for C ₁₉ H ₁₇ N ₆ O ₂ S ₂ : 425.0849; found: 425.0853.

Example 21: 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)-1,3-thiazol4-carboxylic acid

Step A: ethyl 2-[(6-chloro-5-methylpyridazin-3-yl)(methyl)amino]-1,3-thiazol-4-carboxylate.

Sodium hydride (60% dispersion, 240 mg, 6 mmol, 1.18 eq.) under nitrogen atmosphere was slowly added to a solution of 3,6-dichloro-4-methylpyridazine (850 mg, 5.21 mmol, 1.02 eq.) and ethyl 2-(methylamino )-1,3thiazole-4-carboxylate (950 mg, 5.1 mmol, 1 equiv.) in tetrahydrofuran (25 ml) at 0°C. The mixture was stirred for 1 hour at 0°C and for 18 hours at ambient temperature to obtain a dark brown suspension. Saturated aqueous ammonium chloride (75 ml) was added and the mixture was extracted with dichloromethane (250 ml). The extracts were washed with water (2x75 ml) and saline (75 ml). The solution was dried (magnesium sulfate) and concentrated in vacuo. Purification by flash column chromatography (100 g silica gel) eluting with 1% methanol in dichloromethane gave a solid which was triturated with diethyl ether (15 ml), filtered, washed with diethyl ether (2 x 10 ml) and dried in vacuo to give the title product as a pale brown solid (960 mg, 3.07 mmol, 60%).

LC/MS (C ₁₂ H ₁₃ ClN ₄ O ₂ S) 313 [M+H]+; RT 1.17 (LCMS-V-BI).

1H NMR (400 MHz, DMSO) δ 8.04 (s, 1H), 7.86 (s, 1H), 4.30 (q, J=7.1 Hz, 2H), 3.83 (s, 3H), 2.43 (s, 3H), 1.32 (t, J=7.1 Hz, 3H).

Step B: ethyl 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)-1,3

- 101 046336 thiazole-4-carboxylate.

2-Aminobenzothiazole (245 mg, 1.63 mmol, 1.16 eq.) and the product from step A (440 mg, 1.41 mmol, 1 eq.) were added to a solution of tris(dibenzylideneacetone)dipalladium(0) (64.4 mg, 0.07 mmol) under a nitrogen atmosphere , 0.05 eq.) and xantphos (81.4 mg, 0.14 mmol, 0.1 eq.) in 1,4-dioxane (20 ml), and the mixture was stirred. N,N-diisopropylethylamine (0.75 mL, 4.22 mmol, 3 eq.) was added and the mixture was heated in a sealed tube at 150°C for 24 hours. The solution was allowed to cool to ambient temperature and concentrated in vacuo. Purification by flash column chromatography (50 g silica gel), eluting with ethyl acetate, gave a yellow solid, which was triturated with methanol (10 ml), filtered, washed with methanol (15 ml) and dried in vacuo to give the title product as yellow solid (315 mg, 0.74 mmol, 53%).

LC/MS (C ₁₉ H ₁₈ N ₆ O ₂ S ₂ ) 427 [M+H]+; RT 1.32 (LCMS-\-B1).

1H NMR (400 MHz, DMSO) δ 11.23 (br s, 1H), 7.97 (s, 1H), 7.91 (br s, 1H), 7.70 (s, 1H), 7.54 (br s, 1H), 7.39 (td , J=7.7, 1.2 Hz, 1H), 7.22 (t, 1H), 4.30 (q, J=7.1 Hz, 2H), 3.82 (s, 3H), 2.46 (s, 3H), 1.32 (t, J= 7.1 Hz, 3H).

Step C: 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl} (methyl)amino)-1,3-thiazol-4carboxylic acid.

To a solution of the product from step B (200 mg, 0.47 mmol, 1 eq.) in 1,4-dioxane (3 ml) was added lithium hydroxide monohydrate (98.4 mg, 2.34 mmol, 5 eq.) and the mixture was stirred at ambient temperature for 3 hours. Water (10 ml) was added and the solution was filtered through celite, the solids were washed with water (5 ml) and the combined filtrate was acidified with acetic acid to obtain a pale yellow suspension. The solids were collected by filtration, washed with water (20 mL) and dried in vacuo to give the title product as a pale yellow solid (150 mg, 0.34 mmol, 72%).

LC/MS (C ₁₇ H ₁₄ N6O ₂ S ₂ ) 399 [M+H]+; RT 1.1 (LCMS-V-Bi).

1H NMR (400 MHz, DMSOO δ 12.21 (br s, 2H), 7.95-7.84 (m, 2H), 7.71 (s, 1H), 7.54 (d, J=8.0 Hz, 1H), 7.39 (ddd, J= 8.2, 7.3, 1.3 Hz, 1H), 7.22 (td, J=7.6, 1.2 Hz, 1H), 3.82 (s, 3H), 2.47 (s, 3H).

Example 22: 2-({6-[(1,3-benzothiazol-2-yl)amino]-4,5-dimethylpyridazin-3-yl}amino)-5-(3-{4-[3(dimethylamino) propyl]-2-fluorophenoxy}propyl)-1,3-thiazole-4-carboxylic acid

Step A: ethyl 5-(3-{4-[(1E)-3-(dimethylamino)prop-1-en-1-yl]-2-fluorophenoxy}propyl)-2acetamido-1,3-thiazol-4- carboxylate.

Sodium hydride (60% in mineral oil) (1.61 g, 40.3 mmol, 2.34 eq.) was added to a stirred solution of synthesis 6b (4.65 g, 24.1 mmol, 1.4 eq.) in dimethylformamide (70 ml). The reaction mixture was stirred at ambient temperature for 30 min, then cooled in an ice bath and a solution of synthesis product 3u, step C (6.57 g, 17.2 mmol, 1 eq.) in dimethylformamide (30 ml) was slowly added. After 40 minutes, the reaction mixture was allowed to warm to ambient temperature and stirred for 5 hours. The mixture was cooled, acidified with 2N. aqueous HCl and organics were extracted with ethyl acetate and then 3:1 dichloromethane/isopropanol. The combined organics were dried (magnesium sulfate) and the solvent was removed in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 80 g RediSep™ silica gel cartridge) eluting with a gradient of 0-10% methanol in dichloromethane gave the title product as a brown foam (2.26 g, 5.69 mmol, 33%) which used in subsequent stages without further purification. The substance sample was further purified using preparative HPLC (HPLC-VA1 method) to obtain analytical data.

LC/MS (C ₂₂ H ₂₈ FN ₃ O ₄ S) 450 [M+H]+; RT 0.75 (LCMS-\-B1).

1H NMR (400 MHz, DMSO-de) δ 7.25-7.08 (m, 2H), 7.05 (dd, J=8.5, 2.0 Hz, 1H), 6.45 (dt, J=12.0, 1.9 Hz, 1H), 5.69 ( dt, J=12.4, 6.4 Hz, 1H), 4.23 (qd, J=7.1, 2.2 Hz, 2H), 4.09 (t, 2H), 3.27 (t, J=7.5 Hz, 2H), 3.11 (dd, J =6.4, 1.9 Hz, 2H), 2.16 (s, 6H), 2.12 (s, 3H), 2.11-2.06 (m, 2H), 1.27 (t, J=7.1 Hz, 3H).

Step B: ethyl 2-amino-5-(3-{4-[(1E)-3-(dimethylamino)prop-1-en-1-yl]-2-fluorophenoxy}propyl)1,3-thiazol-4 -carboxylate.

To a solution of the product from step A (2.25 g, 4.99 mmol, 1 eq.) in ethanol (40 ml) was added chloride

- 102 046336 hydrogen acid (4 M in 1,4-dioxane; 12.5 ml, 49.9 mmol, 10 eq.), and the mixture was heated at 60°C for 18 hours. The reaction mixture was allowed to cool to ambient temperature and the solvent was removed in vacuum. Purification by automated flash column chromatography (CombiFlash Rf, 40 g RediSep™ silica gel cartridge) eluting with a gradient of 0-20% methanol in dichloromethane provided the crude product. It was dissolved in methanol and loaded onto a methanol-moistened SCX-2 cartridge (10 g). Perform elution with methanol followed by elution with 3.5N. methanolic ammonia and subsequent evaporation gave the target product in the form of a brown gum (2.07 g, 5.07 mmol, 100%).

LC/MS (C ₂₀ H ₂₆ FN ₃ O ₃ S) 408 [M+H]+; RT 0.71 (LCMS-V-BI).

¹ H NMR (400 MHz, DMSOO δ 7.34 (br s, 2H), 7.26-7.15 (m, 3H), 6.74 (d, J=11.9 Hz, 1H), 5.80 (dt, J=12.5, 6.5 Hz, 1H ), 4.16 (q, 2H), 4.09 (t, 2H), 4.03 (dd, 2H), 3.14 (t, J=7.5 Hz, 2H), 2.73 (s, 6H), 2.02 (p, J=6.3 Hz , 2H), 1.24 (t, 3H).

Step C: Ethyl 2-amino-5-(3-{4-[3-(dimethylamino)propyl]-2-fluorophenoxy}propyl)-1,3-thiazol-4carboxylate.

A solution of the product from step B (0.84 g, 2.06 mmol, 1 eq.) in 1:1 ethyl acetate/methanol (40 ml) was loaded into a flask containing platinum oxide hydrate (on the tip of a spatula) under a nitrogen atmosphere. The vessel was evacuated and refilled with nitrogen (x3), then evacuated, created an atmosphere of hydrogen, and shaken at ambient temperature for 72 hours. The mixture was filtered through celite (10 g prefilled cartridge), eluted with methanol and the solvent was removed in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 40 g RediSep™ silica gel cartridge) eluting with a gradient of 0-20% methanol (containing 20% ammonia) in dichloromethane gave the title product as a yellow gum (519 mg, 1.27 mmol, 53%).

LC/MS (C ₂₀ H ₂₈ FN3O ₃ S) 410 [M+H]+; RT 0.70 (LCMS-V-BI).

¹ H NMR (400 MHz, DMSOL ₆ ) δ 7.15-7.03 (m, 2H), 7.02 (s, 2H), 6.95 (dt, J=8.3, 1.3 Hz, 1H), 4.16 (q, J=7.1 Hz, 2H), 4.02 (t, J=6.2 Hz, 2H), 3.12 (dd, J=8.2, 6.8 Hz, 2H), 2.63-2.57 (m, 2H), 2.54 (t, 2H), 2.44 (s, 6H ), 2.05-1.93 (m, 2H), 1.79 (p, J=7.9 Hz, 2H), 1.24 (t, J=7.1 Hz, 3H).

Step D: ethyl 2-({6-[(1,3-benzothiazol-2-yl)amino]-4,5-dimethylpyridazin-3-yl}amino)-5-(3-{4-[3(dimethylamino )propyl]-2-fluorophenoxy}propyl)-1,3-thiazol-4-carboxylate.

Synthesis product 9a (118 mg, 0.41 mmol, 1 eq.), product from step C (200 mg, 0.49 mmol, 1.2 eq.), cesium carbonate (265 mg, 0.81 mmol, 2 equiv.), xanthos (23.6 mg, 0.04 mmol, 0.1 equiv.) and tris(dibenzylideneacetone)dipalladium(0) (18.6 mg, 0.02 mmol, 0.05 equiv.). The vessel was evacuated and purged with nitrogen (x3), and then alpha,alpha,alpha-trifluorotoluene (4 ml) was added. The mixture was bubbled with nitrogen (5 min) and then heated at 170°C for 1 h under microwave irradiation. The mixture was partitioned between ethyl acetate and water, and the organic phase was dried (magnesium sulfate) and concentrated in vacuo. Purification by reverse phase automated flash chromatography (CombiFlash Rf, C18 13 g RediSep column) eluting with 5-95% acetonitrile in water gave the title product as a yellow foam (36 mg, 0.05 mmol, 13%).

LC/MS (C ₃₃ H ₃₈ FN7O ₃ S ₂ ) 664 [M+H]+; RT 1.01 (LCMS-V-BI).

1H NMR (400 MHz, DMSOL ₆ ) δ 10.71 (br s, 1H), 7.81 (d, J=7.8 Hz, 1H), 7.44 (d, J=8.0 Hz, 1H), 7.30 (td, 1H), 7.13 (td, J=7.5, 1.1 Hz, 1H), 7.05-6.95 (m, 2H), 6.91-6.82 (m, 1H), 4.20 (q, J=7.1 Hz, 2H), 4.03 (t, J=6.2 Hz, 2H), 3.28-3.19 (m, 4H), 2.30 (s, 3H), 2.28 (s, 3H), 2.15-2.02 (m, 4H), 2.04 (s, 6H), 1.57 (p, J= 7.4 Hz, 2H), 1.23 (t, J=7.1 Hz, 3H).

Step E: 2-({6-[(1,3-benzothiazol-2-yl)amino]-4,5-dimethylpyridazin-3-yl}amino)-5-(3-{4-[3(dimethylamino) propyl]-2-fluorophenoxy}propyl)-1,3-thiazole-4-carboxylic acid.

To a solution of the product from step D (57.1 mg, 0.09 mmol, 1 eq.) in 1,4-dioxane (5 ml) was added lithium hydroxide monohydrate (14.4 mg, 0.34 mmol, 4 eq.) and the mixture was refluxed at for 5.5 hours. The mixture was allowed to cool to ambient temperature and the solvent was removed in vacuo. The residue was triturated with water and the solids were collected by filtration. The solids were further triturated with diethyl ether, then collected by filtration and dried in vacuo to give the title product as a brown solid (19.1 mg, 0.03 mmol, 35%).

HRMS-ERI (m/z) [M+H]+, calculated for C ₃₁ H ₃₅ FN ₇ O ₃ S ₂ : 636.2221; found: 636.2224.

- 103 046336

Example 23: 2-[[1-(1,3-Benzothiazol-2-ylamino)-6,7-dihydro-5H-cyclopenta^]pyridazin-4yl]amino]thiazol-4-carboxylic acid

Step A: 1,4-dibromo-6,7-dihydro-5H-cyclopenta^]pyridazine.

A mixture of 1.86 g of 2,3,6,7-tetrahydro-1H-cyclopenta^]pyridazin-1,4(5H)-dione (12.3 mmol, 1 eq.) and 10.55 g (36.8 mmol, 3 eq.) POBr ₃ in 50 ml of 1,2-dichloroethane were kept under reflux for 3 hours. After cooling to room temperature. and concentration, the residue was treated with 100 g of ice, the pH was adjusted to 7 by adding conc. NaHCO ₃ and the solid was filtered off to obtain 1.88 g (55%) of the desired product.

Step B: 4-bromo-6,7-dihydro-5H-cyclopenta^]pyridazin-1-amine.

A mixture of 1.88 g of the product from stage A (6.8 mmol) in 25 ml of a 30% NH3 solution in water was kept at 130°C for 18 hours. After concentration, the residue was added to EtOAc, washed with a 10% NaOH solution, dried over _MgSO4 , filtered and concentrated to obtain 1.28 g (88%) of the target product.

LC-MS-ESI (m/z): [M+H]+, calculated for C ₇ H ₉ BrM ₃ : 214, 216; found: 214, 216.

Step C: Na(4-bromo-6,7-dihydro-5H-cyclopenta^]pyridazin-1-yl)-1,3-benzothiazol-2-amine.

A mixture of 214 mg of the product from step B (1 mmol, 1 eq.), 321 mg of 2-bromo-1,3-benzothiazole (1.50 mmol, 1.5 eq.) and 160 mg of sodium hydride (60% suspension) (4 mmol, 4 eq.) in 25 ml of 1,4-dioxane was kept under reflux for 1 hour. After cooling and quenching the reaction mixture with methanol, the mixture was concentrated, treated with water, the product was filtered and dried. The solid was triturated with EtOAc and dried to obtain 275 mg (79%) of the expected product.

LC-MS-ESI (m/z): [M+H] ⁺ , calculated for C ₁₄ H ₁₂ BrN ₄ S: 347, 349; found: 347, 349.

Step D: Ethyl 2-[[1-(1,3-benzothiazol-2-ylamino)-6,7-dihydro-5H-cyclopenta^]pyridazin-4yl]amino]thiazol-4-carboxylate.

A mixture of 270 mg of the product from step C (0.778 mmol, 1 eq.), 134 mg of ethyl 2-aminothiazole-4-carboxylate (0.78 mmol, 1 eq.), 29 mg of Pd ₂ (dba) ₃ (0.032 mmol, 0.04 eq. ), 45 mg of xanthos (0.078 mmol, 0.1 eq.) and 507 mg of Cs ₂ CO ₃ (1.56 mmol, 2 eq.) in 10 ml of trifluoromethylbenzene were kept at 200°C for 0.5 h. The reaction mixture was concentrated and purified using a preparative HPLC to obtain 97 mg (28%) of the desired product.

LC-MS-ESI (m/z): [M+H]+, calculated for C20H19N6O2S2: 439.5; found: 439.

Step E: 2-[[1-(1,3-benzothiazol-2-ylamino)-6,7-dihydro-5H-cyclopenta^]pyridazin-4yl]amino]thiazol-4-carboxylic acid.

A mixture of 90 mg of the product from step D (0.22 mmol, 1 eq.), 18 mg of LiOHxH ₂ O (0.43 mmol, 2 eq.) and 13 drops of water in 5 ml of 1,4-dioxane was kept under reflux for 2 h. The reaction mixture was concentrated and purified using preparative HPLC to obtain 54 mg (64%) of the target product.

MSHR-ERI (m/z): [M+H]+, calculated for C ₁₈ H ₁₅ N ₆ O ₂ S ₂ : 411.0697; found: 411.0685.

Example 24: 6-[[4-(1,3-Benzothiazol-2-ylamino)-5,6,7,8-tetrahydrophthalazin-1-yl]amino]pyridine-2carboxylic acid

Step A: ethyl 6-[[4-(1,3-benzothiazol-2-ylamino)-5,6,7,8-tetrahydrophthalazin-1-yl]amino]pyridine 2-carboxylate.

A mixture of 245 mg of the compound of example 17, stage B (0.68 mmol, 1 eq.), 113 mg ethyl 6-aminopyridine-2-carboxylate (0.68 mmol, 1 eq.), 25 mg Pd ₂ (dba) ₃ (0.027 mmol, 0.04 eq. ), 40 mg of xanthos (0.069 mmol, 0.1 eq.) and 443 mg of Cs ₂ CO ₃ (1.36 mmol, 2 eq.) in 10 ml of trifluoromethylbenzene were kept at 200°C for 1.5 h. The reaction mixture was concentrated and purified using a preparative HPLC to obtain 45 mg (15%) of the desired product.

- 104 046336

LC-MS-ESI (m/z): [M+H]+, calculated for C23H23N6O2S: 447; found: 447.

Step B: 6-[[4-(1,3-benzothiazol-2-ylamino)-5,6,7,8-tetrahydrophthalazin-1-yl]amino]pyridine-2-carboxylic acid.

A mixture of 45 mg of the product from stage A (0.11 mmol, 1 eq.), 8.5 mg of LiOHxH ₂ O (0.20 mmol, 2 eq.) and 10 drops of water in 10 ml of 1,4-dioxane was kept under reflux for 2 h. The reaction mixture was concentrated and purified using preparative HPLC to obtain 21 mg (50%) of the target product.

MSHR-ERI (m/z): [M+H]+, calculated for C21H19N6O2S: 419.1290; found: 419.1294.

Example 25: 6-[[4-(1,3-Benzothiazol-2-ylamino)-6,7-dihydro-5H-cyclopenta^]pyridazin-1yl]amino]pyridine-2-carboxylic acid

Step A: 6-[[4-(1,3-benzothiazol-2-ylamino)-6,7-dihydro-5H-cyclopenta^]pyridazin-1yl]amino]pyridin-2-carboxylate.

A mixture of 290 mg of the compound of example 23, step C (0.84 mmol, 1 eq.), 139 mg of ethyl 6-aminopyridine-2-carboxylate (0.84 mmol, 1 eq.), 31 mg of Pd ₂ (dba) ₃ (0.034 mmol, 0.04 eq. ), 48 mg (0.083 mmol, 0.1 eq.) of xantphos and 545 mg of Cs ₂ CO ₃ (1.67 mmol, 2 eq.) in 10 ml of trifluoromethylbenzene were kept at 200°C for 1.5 h. The reaction mixture was concentrated and purified using a preparative HPLC to obtain 70 mg (19.4%) of the target product.

LC-MS-ESI (m/z): [M+H]+, calculated for C ₂₂ H ₂₁ N ₆ O ₂ S: 433.5; found: 433 and 431 [M-N]'.

Step B: 6-[[4-(1,3-benzothiazol-2-ylamino)-6,7-dihydro-5H-cyclopenta^]pyridazin-1yl]amino]pyridine-2-carboxylic acid.

A mixture of 70 mg of the product from stage A (0.16 mmol, 1 eq.), 14 mg of LiOHxH ₂ O (0.33 mmol, 2 eq.) and 10 drops of water in 10 ml of 1,4-dioxane was kept under reflux for 2 h. The reaction mixture was concentrated and purified using preparative HPLC to obtain 24 mg (31%) of the target product.

MSHR-ERI (m/z): [M+H]+, calculated for C ₂₀ H ₁₇ N ₆ O ₂ S: 405.1134; found: 405.1123.

Example 26: 2-({6-[(1,3-benzothiazol-2-yl)amino]-4,5-dimethylpyridazin-3-yl} (methyl)amino)-1,3thiazol-4-carboxylic acid

Step A: ethyl 2-[(6-chloro-4,5-dimethylpyridazin-3-yl)(methyl)amino]-1,3-thiazol-4-carboxylate.

Sodium hydride (60% dispersion, 250 mg, 6.25 mmol, 1.16 eq.) under nitrogen atmosphere was slowly added to a solution of 3,6-dichloro-4,5-dimethylpyridazine (950 mg, 5.37 mmol, 1 eq.) and ethyl 2- (methylamino)-1,3thiazole-4-carboxylate (1.2 g, 6.44 mmol, 1.2 eq.) in 1,4-dioxane (20 ml) at 0°C. The mixture was stirred for 1 hour at ambient temperature, then at 60°C for 2 hours. Saturated aqueous ammonium chloride (75 ml) was added and the mixture was extracted with ethyl acetate (150 ml). The organic extract was washed with water (2x75 ml), brine (75 ml), dried (magnesium sulfate) and concentrated in vacuo. Purification by column chromatography (100 g silica gel) eluting with ethyl acetate gave the title product as a brown gum (715 mg, 2.19 mmol, 41%).

LC/MS (C ₁₃ H ₁₅ ClN ₄ O ₂ S) 327 [M+H]+; RT 1.09 (ZhKhMS-U-V1).

1H NMR (400 MHz, CDC13) δ 7.56 (s, 1H), 4.34 (q, J=7.2 Hz, 2H), 3.66 (s, 3H), 2.43 (s, 3H), 2.23 (s, 3H), 1.36 (t, J=7.2 Hz, 3H).

Step B: ethyl 2-({6-[(1,3-benzothiazol-2-yl)amino]-4,5-dimethylpyridazin-3-yl}(methyl)amino)-1,3thiazol-4-carboxylate.

2-Aminobenzothiazole (250 mg, 1.66 mmol, 1.21 eq.) and the product from step A (500 mg, 1.38 mmol, 1 eq.) were added to a solution of tris(dibenzylideneacetone)dipalladium(0) (63.1 mg, 0.07 mmol) under a nitrogen atmosphere , 0.05 eq.) and xanthos (79.7 mg, 0.14 mmol, 0.1 eq.) in 1,4-dioxane (20 ml), and the mixture was stirred

- 105 046336 li. N,N-diisopropylethylamine (0.75 mL, 4.13 mmol, 3 eq.) was added and the mixture was heated in a sealed tube at 150°C for 24 hours. The solution was allowed to cool to ambient temperature and concentrated in vacuo. Purification by flash column chromatography (50 g silica gel) eluting with 1% methanol in dichloromethane gave a dark yellow resin which was triturated with diethyl ether (15 ml), filtered, washed with diethyl ether (15 ml) and dried in vacuo to obtain the desired product as a pale yellow solid (365 mg, 0.83 mmol, 60%).

LC/MS (C ₂₀ H ₂₀ N6O ₂ S ₂ ) 441 [M+H]+; RT 1.26 (LCMS-V-BI).

¹ H NMR (400 MHz, DMSO-с1 ₆ ) δ 11.22 (br s, 1H), 7.87 (br s, 1H), 7.76 (s, 1H), 7.56 (br s, 1H), 7.40 (t, J= 7.5 Hz, 1H), 7.23 (t, J=7.7 Hz, 1H), 4.25 (q, J=7.1 Hz, 2H), 3.49 (s, 3H), 2.41 (s, 3H), 2.19 (s, 3H) , 1.28 (t, J=7.1 Hz, 3H).

Step C: 2-({6-[(1,3-benzothiazol-2-yl)amino]-4,5-dimethylpyridazin-3-yl}(methyl)amino)-1,3thiazol-4-carboxylic acid.

To a solution of the product from step B (200 mg, 0.45 mmol, 1 eq.) in a 1:3 mixture of water/1,4-dioxane (4 ml) was added lithium hydroxide monohydrate (95.3 mg, 2.27 mmol, 5 eq.), and the mixture was stirred at ambient temperature for 18 hours. Water (10 ml) was added and the solution was filtered through celite, the solids were washed with water (5 ml) and the combined filtrate was acidified with acetic acid to obtain a pale yellow suspension. The solids were collected by filtration, washed with water (20 mL) and dried in vacuo to give the title product as a pale yellow solid (150 mg, 0.36 mmol, 80%).

MSHR-ERI (m/z) [M—H]—, calculated for C18H15N6O2S2: 411.0703; found: 411.0689.

Example 27: 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-(hydroxymethyl)-4-methylpyridazin-3-yl}amino)1,3-thiazol-4-carboxylic acid

Step A: ethyl 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-(methoxymethyl)-4-methylpyridazin-3yl}amino)-1,3-thiazol-4-carboxylate.

Synthesis product 9jb (169 mg, 0.53 mmol, 1 eq.), ethyl 2-amino-1,3-thiazol-4-carboxylate (109 mg, 0.63 mmol, 1.2 eq.) was loaded into a microwave-dried vessel for microwave treatment. , cesium carbonate (240 mg, 0.74 mmol, 1.4 eq.), xanthos (30.5 mg, 0.05 mmol, 0.1 eq.) and tris(dibenzylideneacetone) dipalladium(0) (24.1 mg, 0.03 mmol, 0.05 eq.). The vessel was evacuated and purged with nitrogen (x3), and then alpha,alpha,alpha-trifluorotoluene (10 ml) was added. The mixture was bubbled with nitrogen (10 min) and then heated at 180°C for 1 h under microwave irradiation. The mixture was concentrated in vacuo and purified by automated flash column chromatography (CombiFlash Rf, 12 g RediSep™ silica gel cartridge) eluting with a gradient of 0-5% methanol in dichloromethane to give the title product (138 mg, 0.30 mmol, 57%).

LC/MS (C20H20N6O3S2) 457 [M+H]+; RT 1.17 (LCMS-V-BI).

¹ H NMR (400 MHz, DMSO^) δ 12.25 (br s, 1H), 11.07 (br s, 1H), 10.63 (br s, 1H), 7.96 (s, 1H), 7.87 (s, 1H), 7.49 (br s, 1H), 7.38 (t, J=7.5 Hz, 1H), 7.20 (t, 1H), 4.78 (s, 2H), 4.30 (q, J=7.1 Hz, 2H), 3.38 (s, 3H ), 2.43 (s, 3H), 1.33 (t, J=7.1 Hz, 3H).

Step B: ethyl 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-(hydroxymethyl)-4-methylpyridazin-3yl}amino)-1,3-thiazol-4-carboxylate.

To a solution of the product from step A (55 mg, 0.12 mmol, 1 eq.) in dichloromethane (2 ml) at 0°C, boron tribromide (0.6 ml, 1 M, 0.6 mmol, 5 eq.) was added and the mixture was stirred at environment for 1 hour. The reaction mixture was quenched with methanol and concentrated in vacuo. The solid was added to methanol and refluxed for 30 minutes, then allowed to cool to ambient temperature. Purification by reverse phase automated flash column chromatography (CombiFlash Rf, C18 4.3 g RediSep column) eluting with a 5-95% acetonitrile gradient gave crude product (53 mg), which was carried to the next step without further purification.

Step C: 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-(hydroxymethyl)-4-methylpyridazin-3-yl}amino)1,3-thiazol-4-carboxylic acid .

To a solution of the product from step B (53.3 mg, 0.12 mmol, 1 eq.) in 1,4-dioxane (5 ml) was added a 2 M aqueous solution of lithium hydroxide (0.24 ml, 0.24 mmol, 2 eq.), and the mixture was heated with reflux for 1 hour. The reaction mixture was allowed to cool to ambient temperature and

- 106 046336 centered in vacuum. The residue was added to a minimal amount of water, acidified to pH 6 with 1 M aqueous hydrochloric acid, and allowed to stir for 10 minutes. The solvent was removed in vacuo and the residue was triturated with 5:1 isopropanol/water. The suspension was stirred for 30 min, then the solid was collected by filtration, washed with isopropanol and dried in vacuo to give the title product as a brown solid (16 mg, 0.04 mmol, 32%).

MSVR-ERI (m/z) [MH] ^- , calculated for C ₁₇ H ₁₃ N ₆ O ₃ S ₂ : 413.0496; found: 413.0499.

Example 28: 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-(methoxymethyl)-4-methylpyridazin-3-yl}amino)1,3-thiazol-4-carboxylic acid

To a solution of the product from Example 27, Step A (59 mg, 0.13 mmol, 1 eq.) in 1,4-dioxane (5 ml) was added a 2 M aqueous solution of lithium hydroxide (0.26 ml, 0.26 mmol, 2 eq.), and the mixture was refluxed for 1 hour. The mixture was allowed to cool to ambient temperature and concentrated in vacuo. The residue was added to a minimal amount of water, acidified to pH 6 with 1 M aqueous hydrochloric acid and allowed to stir for 10 minutes. The solvent was removed in vacuo and the residue was triturated with 5:1 isopropanol/water. The suspension was stirred for 30 min, then the solids were collected by filtration, washed with isopropanol and dried in vacuo to give the title product as an orange solid (18.5 mg, 0.04 mmol, 33%).

MSVR-ERI (m/z) [MH] ^- , calculated for C ₁₈ H ₁₅ N ₆ O ₃ S ₂ : 427.0653; found: 427.0650.

Example 29: 2-({6-[(1,3-benzothiazol-2-yl)amino]-4-(methoxymethyl)-5-methylpyridazin-3-yl}amino)1,3-thiazol-4-carboxylic acid

Step A: methyl 2-({6-[(1,3-benzothiazol-2-yl)amino]-4-(methoxymethyl)-5-methylpyridazin-3yl} amino)-1,3-thiazol-4-carboxylate.

Synthesis product 9ja (85 mg, 0.26 mmol, 1 eq.), ethyl 2-amino-1,3-thiazol-4-carboxylate (59.3 mg, 0.34 mmol, 1.3 eq.) was loaded into a microwave-dried vessel for microwave treatment. , cesium carbonate (121 mg, 0.37 mmol, 1.4 eq.), xantphos (7.67 mg, 0.01 mmol, 0.05 eq.) and tris(dibenzylideneacetone) dipalladium(0) (24.3 mg, 0.03 mmol, 0.1 eq.). The vessel was evacuated and purged with nitrogen (x3), and then alpha,alpha,alpha-trifluorotoluene (10 ml) was added. The mixture was bubbled with nitrogen (10 min) and then heated at 180°C for 1 h under microwave irradiation. The mixture was concentrated in vacuo and purified by automated flash column chromatography (CombiFlash Rf, 12 g RediSep™ silica gel cartridge) eluting with a gradient of 0-5% methanol in dichloromethane to give the title product as an orange glassy solid (82.5 mg, 0.18 mmol, 68%).

LC/MS (C ₁₉ H ₁₈ N ₆ O ₃ S ₂ ) 443 [M+H]+; RT 1.26 (LCMS-V-BI).

1H NMR (400 MHz, DMSO) δ 10.78 (br s, 1H), 7.98 (s, 1H), 7.42-7.34 (m, 2H), 7.25-7.14 (m, 2H), 4.77 (s, 2H), 3.83 (s, 3H), 3.39 (s, 3H), 2.44 (s, 3H).

Step B: 2-({6-[(1,3-benzothiazol-2-yl)amino]-4-(methoxymethyl)-5-methylpyridazin-3-yl}amino)1,3-thiazol-4-carboxylic acid .

To a solution of the product from step A (82.5 mg, 0.18 mmol, 1 eq.) in 1,4-dioxane (5 ml) was added a 2 M aqueous solution of lithium hydroxide (0.18 ml, 0.36 mmol, 2 eq.), and the mixture was heated with reflux for 1 hour. The reaction mixture was allowed to cool to ambient temperature and concentrated in vacuo. The residue was added to a minimal amount of water, acidified to pH 6 with 1 M aqueous hydrochloric acid, and allowed to stir for 10 min.

- 107 046336

The solvent was removed in vacuo and the residue was triturated with 5:1 isopropanol/water. Solids were collected by filtration, washed with isopropanol (2x4 ml) and dried in vacuum. Purification by reverse phase HPLC (HPLC-U-A2 method) provided the title product as a yellow solid (22 mg, 0.05 mmol, 28%).

HRMS-ERI (m/z) [M+H]+, calculated for C ₁₈ H ₁₇ N ₆ O ₃ S ₂ : 429.0798; found: 429.0800.

Example 30: 2-({6-[(1,3-benzothiazol-2-yl)amino]-4-(ethoxymethyl)-5-methylpyridazin-3-yl}amino)1,3-thiazol-4-carboxylic acid

Step A: ethyl 2-({6-[(1,3-benzothiazol-2-yl)amino]-4-(ethoxymethyl)-5-methylpyridazin-3yl}amino)-1,3-thiazol-4-carboxylate.

Synthesis product 9ka (113 mg, 0.34 mmol, 1 eq.), ethyl 2-amino-1,3-thiazol-4-carboxylate (69.7 mg, 0.4 mmol, 1.2 eq.) was loaded into a microwave-dried vessel for microwave treatment. , cesium carbonate (154 mg, 0.47 mmol, 1.4 eq.), xanthos (19.5 mg, 0.03 mmol, 0.1 eq.) and tris(dibenzylideneacetone) dipalladium(0) (15.5 mg, 0.02 mmol, 0.05 eq.). The vessel was evacuated and purged with nitrogen (x3), and then alpha,alpha,alpha-trifluorotoluene (7 ml) was added. The mixture was bubbled with nitrogen (10 min) and then heated at 180°C for 1 h under microwave irradiation. The mixture was concentrated in vacuo and purified by automated flash column chromatography (CombiFlash Rf, 24 g RediSep™ silica gel cartridge) eluting with a gradient of 0-100% ethyl acetate in isoheptane to give the title product (47 mg, 0.1 mmol, 30%).

LC/MS (C ₂₁ H ₂₂ N ₆ O ₃ S ₂ ) 471 [M+H]+; RT 1.36 (LCMS-V-BI).

1H NMR (400 MHz, CDCl ₃ ) δ 7.82 (s, 1H), 7.73 (d, J=35.3 Hz, 3H), 7.43 (t, 1H), 7.28-7.23 (m, 1H), 4.72 (s, 2H ), 4.45 (q, J=7.1 Hz, 2H), 3.68 (q, J=7.0 Hz, 2H), 2.47 (s, 3H), 1.45 (t, J=7.1 Hz, 3H), 1.35 (t, J =7.0 Hz, 3H).

Step B: 2-({6-[(1,3-benzothiazol-2-yl)amino]-4-(ethoxymethyl)-5-methylpyridazin-3-yl} amino)-1,3 thiazol-4-carboxylic acid .

To a solution of the product from step A (49 mg, 0.1 mmol, 1 eq.) in 1,4-dioxane (5 ml) was added a 2 M aqueous solution of lithium hydroxide (0.1 ml, 0.21 mmol, 2 eq.), and the mixture was heated with reflux for 2 hours. The mixture was allowed to cool to ambient temperature and concentrated in vacuo. The residue was added to a minimal amount of water, acidified to pH 6 with 1 M aqueous hydrochloric acid, and allowed to stir for 10 min. The sample was triturated with 5:1 isopropanol/water and the solids were collected by filtration, washed with isopropanol and dried in vacuo to give the title product as a brown solid (23.9 mg, 0.05 mmol, 52%).

MSVR-ERI (m/z) [MH] ^- , calculated for C ₁₉ H ₁₇ N ₆ O ₃ S ₂ : 441.0809; found: 441.0803.

Example 31: 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-cyclopropylpyridazin-3-yl}(methyl)amino)-1,3thiazol-4-carboxylic acid

To a solution of synthesis product 11d, step B (249 mg, 0.55 mmol, 1 equiv.) in 1,4-dioxane (10 ml) was added a 2 M aqueous solution of lithium hydroxide (0.55 ml, 1.1 mmol, 2 equiv.), and the mixture reflux for 3 hours. The reaction mixture was allowed to cool to ambient temperature and concentrated in vacuo. Purification by reverse phase automated flash chromatography (CombiFlash Rf, C18 43 g RediSep column) eluting with a gradient of 5-95% acetonitrile in water gave the title product as a yellow powder (33 mg, 0.08 mmol, 14%) [in form of lithium salt].

HRMS-ERI (m/z) [M+H]+, calculated for C ₁₉ H ₁₇ N ₆ O ₂ S ₂ : 425.0849; found: 425.0851.

Example 32: 5-{1-[(adamantan-1-yl)methyl]-5-methyl-1H-pyrazol-4-yl}-2-({6-[(1,3-benzothiazol-2

- 108 046336 yl)amino]pyridazin-3-yl}amino)-1,3-thiazol-4-carboxylic acid

Step A: ethyl 5-['1-[(adamantan-1-yl)methyl]-5-methyl-1H-pyrazol-4-yl}-2-[(6-['[(^)-3-[ '[2(trimethylsilyl)ethoxy]methyl}-2,3-dihydro-1,3-benzothiazol-2-ylidene]amino}pyridazin-3-yl)amino]1,3-thiazol-4-carboxylate.

To a solution of synthesis product 11f (64 mg, 0.11 mmol, 1 eq.), synthesis product 10a (45.0 mg, 0.13 mmol, 1.2 eq.) and potassium carbonate (43.7 mg, 0.32 mmol, 3 eq.) in a 3:1 mixture tetrahydrofuran/water (4 ml), [1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium(II) (7.71 mg, 0.01 mmol, 0.1 eq.) was added. The mixture was bubbled with nitrogen (10 min) and then heated at 120°C for 1 h under microwave irradiation. The mixture was diluted with ethyl acetate, then washed successively with water and brine. The organic extract was dried (magnesium sulfate) and the solvent was removed in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 12 g RediSep™ silica gel cartridge) eluting with a gradient of 0-50% ethyl acetate in isoheptane provided the title product as a cream solid (39.2 mg, 0.05 mmol, 49%) .

LC/MS (C38H48N8OsSiS2) 757 [M+H]+; RT 1.64 (ZhKhMS-U-V1).

Step B: ethyl 5-{1-[(adamantan-1-yl)methyl]-5-methyl-1H-pyrazol-4-yl}-2-({6-[(1,3-benzothiazol-2yl)amino ]pyridazin-3-yl}amino)-1,3-thiazol-4-carboxylate.

A solution of the product from step A (39 mg, 0.05 mmol, 1 eq.) in dichloromethane (2 ml) was cooled to 0°C and then trifluoroacetic acid (0.3 ml, 4.03 mmol, 78.1 eq.) was added. The mixture was allowed to warm to ambient temperature and stirred for 18 hours. The mixture was diluted with dichloromethane and washed successively with saturated aqueous sodium bicarbonate, water and brine. The organic phase was dried (magnesium sulfate) and then the solvent was removed in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 12 g RediSep™ silica gel cartridge) eluting with a gradient of 0-7% methanol in dichloromethane provided the title product as a cream solid (10.5 mg, 0.02 mmol, 33%) .

LC/MS (C ₃₂ H3 ₄ N ₈ O ₂ S ₂ ) 627 [M+H] ⁺ ; RT 1.38 (LCMS-V-BT).

Ή NMR (400 MHz, DMSO) δ 11.87 (br s, 1H), 11.67 (br s, 1H), 7.93 (d, J=7.8 Hz, 1H), 7.64 (d, 1H), 7.57 (s, 1H) , 7.51 (d, J=27.8 Hz, 1H), 7.44-7.33 (m, 2H), 7.21 (t, J=7.6 Hz, 1H), 4.17 (q, J=7.1 Hz, 2H), 3.80 (s, 2H), 2.23 (s, 3H), 1.98 (s, 3H), 1.72-1.56 (m, 12H), 1.20 (t, J=7.1 Hz, 3H).

Step C: 5-{1-[(adamantan-1-yl)methyl]-5-methyl-1H-pyrazol-4-yl}-2-({6-[(1,3-benzothiazol-2yl)amino] pyridazin-3-yl}amino)-1,3-thiazol-4-carboxylic acid.

To a solution of the product from step B (10.5 mg, 0.02 mmol, 1 eq.) in 1,4-dioxane (2 ml) was added lithium hydroxide monohydrate (3.51 mg, 0.08 mmol, 5 eq.), and the mixture was refluxed at during the night. The reaction mixture was allowed to cool to ambient temperature and the solvent was removed in vacuo. The residue was triturated with water and the solids were collected by filtration and dried in vacuo to give the title product as a beige solid (7.2 mg, 0.01 mmol, 71%).

MSHR-ERI (m/z) [M+H] ⁺ , calculated for C3 ₀ H ₃₁ N ₈ O ₂ S ₂ : 599.2006; found: 599.2037.

Example 33: 5-{1-[(adamantan-1-yl)methyl]-5-methyl-1H-pyrazol-4-yl}-2-({6-[(1,3-benzothiazol-2yl)amino] -4,5-dimethylpyridazin-3-yl}amino)-1,3-thiazole-4-carboxylic acid

Step A: ethyl 5-{1-[(adamantan-1-yl)methyl]-5-methyl-1H-pyrazol-4-yl}-2-[(4,5-dimethyl-6-{[(2Z) -3{[2-(trimethylsilyl)ethoxy]methyl}-2,3-dihydro-1,3-benzothiazol-2-ylidene]amino}pyridazin-3yl)amino]-1,3-thiazol-4-carboxylate.

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To a mixture of synthesis product 11a (99 mg, 0.16 mmol, 1 eq.), synthesis product 10a (66.6 mg, 0.19 mmol, 1.2 eq.) and potassium carbonate (64.6 mg, 0 mol, 3 eq.) in a 3:1 mixture tetrahydrofuran/water (4 ml), [1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium(II) (11.4 mg, 0.02 mmol, 0.1 eq.) was added. The mixture was bubbled with nitrogen (10 min) and then heated at 120°C for 1 h under microwave irradiation. The reaction mixture was diluted with ethyl acetate, then washed successively with water and brine. The organic extract was dried (magnesium sulfate), then the solvent was removed in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 12 g RediSep™ silica gel cartridge) eluting with a gradient of 0-50% ethyl acetate in isoheptane gave the title product as a yellow glass (65.5 mg, 0.08 mmol, 54%) .

LC/MS (C ₄₀ H ₅₂ N ₈ O ₃ SiS ₂ ) 785 [M+H]+; RT 1.82 (LCMS-V-BI).

¹ HNMR (400 MHz, DMSO^) δ 10.93 (br s, 1H), 7.78 (d, J=7.6 Hz, 1H), 7.54 (s, 1H), 7.46-7.38 (m, 2H), 7.21 (dt, 1H), 5.84 (s, 2H), 4.17 (q, J=7.1 Hz, 2H), 3.78 (s, 2H), 3.71 (t, 2H), 2.40 (s, 3H), 2.36 (s, 3H), 2.20 (s, 3H), 1.96 (s, 3H), 1.71-1.55 (m, 12H), 1.17 (t, 3H), 0.91 (t, 2H), 0.00 (s, 9H).

Step B: ethyl 5-{1-[(adamantan-1-yl)methyl]-5-methyl-1H-pyrazol-4-yl}-2-({6-[(1,3-benzothiazol-2yl)amino ]-4,5-dimethylpyridazin-3-yl} amino)-1,3-thiazol-4-carboxylate.

A solution of the product from Step A (65.5 mg, 0.08 mmol, 1 eq.) in dichloromethane (4 mL) was cooled in an ice bath and then trifluoroacetic acid (0.4 mL, 5.37 mmol, 64 eq.) was added. The mixture was allowed to warm to ambient temperature and stirred for 18 hours. The reaction mixture was diluted with dichloromethane and washed successively with saturated aqueous sodium bicarbonate, water and brine. The organics were dried (magnesium sulfate) and the solvent was removed in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 12 g RediSep™ silica gel cartridge) eluting with a gradient of 0-5% methanol in dichloromethane gave the title product as a yellow glass (34.2 mg, 0.05 mmol, 63%) .

LC/MS (C ₃₄ H ₃₈ N ₈ O ₂ S ₂ ) 655 [M+H] ⁺ ; RT 1.47 (LCMS-V-BI).

1H NMR (400 MHz, DMSO^) δ 10.94 (br s, 1H), 7.88 (br s, 1H), 7.64 (br s, 1H), 7.55 (s, 1H), 7.36 (t, 1H), 7.22- 7.13 (m, 1H), 4.18 (q, J=7.1 Hz, 2H), 3.80 (s, 2H), 2.38 (s + s, 6H), 2.21 (s, 3H), 1.99 (d, J=8.2 Hz , 3H), 1.73-1.54 (m, 12H), 1.18 (t, J=7.1 Hz, 3H).

Step C: 5-{1-[(adamantan-1-yl)methyl]-5-methyl-1H-pyrazol-4-yl}-2-({6-[(1,3-benzothiazol-2yl)amino] -4,5-dimethylpyridazin-3-yl}amino)-1,3-thiazol-4-carboxylic acid.

To a solution of the product from step B (34.2 mg, 0.05 mmol, 1 eq.) in 1,4-dioxane (5 ml) was added lithium hydroxide monohydrate (22 mg, 0.52 mmol, 10 eq.) and the mixture was refluxed at for 18 hours. The reaction mixture was allowed to cool to ambient temperature, then the solvent was removed in vacuo. The residue was triturated with water and the solids were collected by filtration and dried in vacuo to give the title product as a yellow solid (28 mg, 0.04 mmol, 86%) [as lithium salt].

HRMS-ERI (m/z) [M+H] ⁺ , calculated for C ₃₂ H ₃₅ N ₈ O ₂ S ₂ : 627.2319; found: 627.2357.

Example 34: 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-(propan-2-yl)pyridazin-3-yl}(methyl)amino)-1,3thiazol-4 -carboxylic acid

Step A: ethyl 2-{[6-chloro-5-(propan-2-yl)pyridazin-3-yl](methyl)amino}-1,3-thiazol-4carboxylate.

To a solution of synthesis product 7c (300 mg, 1.57 mmol, 1 eq.) and ethyl 2-(methylamino)-1,3-thiazol-4carboxylate (292 mg, 1.57 mmol, 1 eq.) in tetrahydrofuran (25 ml) in atmosphere nitrogen, sodium hydride (60% in mineral oil; 75.4 mg, 1.88 mmol, 1.2 eq.) was added slowly at 0°C, and the mixture was stirred at 0°C for 1 hour, then at ambient temperature for 18 hours. saturated aqueous ammonium chloride solution (20 ml), and the mixture was extracted with dichloromethane (75 ml). The organic extracts were washed successively with water (2x50 ml) and brine (50 ml), then dried (magnesium sulfate) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 24 g RediSep™ silica gel cartridge) eluting with a gradient of 0-50% ethyl acetate in isoheptane provided the title product as a white crystalline solid (340 mg, 1 mmol, 64% ).

LC/MS (C ₁₄ H ₁₇ ClN ₄ O ₂ S) 341 [M+H] ⁺ ; RT 1.31 (LCMS-V-BI).

1H NMR (400 MHz, DMSO) δ 8.05 (s, 1H), 7.67 (s, 1H), 4.30 (q, J=7.1 Hz, 2H), 3.88 (s, 3H), 3.20 (hept, J=6.9 Hz , 1H), 1.32 (dt, J=7.1, 3.7 Hz, 9H).

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Step B: ethyl 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-(propan-2-yl)pyridazin-3-yl}(methyl)amino)1,3-thiazole -4-carboxylate.

To a solution of the product from step A (340 mg, 1 mmol, 1 eq.) and 2-aminobenzothiazole (180 mg, 1.2 mmol, 1.2 eq.) in 1,4 dioxane (15 ml) under nitrogen atmosphere was added xanthos (115 mg, 0.2 mmol, 0.2 equiv.) and tris(dibenzylideneacetone)dipalladium(0) (91.4 mg, 0.1 mmol, 0.1 equiv.). N,Ndiisopropylethylamine (0.52 mL, 2.99 mmol, 3 eq.) was added and the mixture was heated in a sealed tube at 150°C for 18 hours. The mixture was allowed to cool to ambient temperature and the solvent was removed in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 24 g RediSep™ silica gel cartridge) eluting with a gradient of 0-3% methanol in dichloromethane provided the title product as a yellow solid (110 mg, 0.24 mmol, 24%) .

LC/MS (C ₂₁ H ₂₂ N ₆ O ₂ S ₂ ) 455 [M+H]+; RT 1.44 (LCMS-V-BI).

1H NMR (400 MHz, DMSOO δ 12.40 (br s, 1H), 7.97 (s, 1H), 7.79 (br s, 1H), 7.55 (s, 1H), 7.37 (br s + t, 2H), 7.19 ( br s, 1H), 4.30 (q, J=7.1 Hz, 2H), 3.87 (s, 3H), 3.57 (p, J=6.8 Hz, 1H), 1.40-1.27 (m, 9H).

Step C: 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-(propan-2-yl)pyridazin-3-yl} (methyl)amino)-1,3thiazol-4 -carboxylic acid.

To a stirred solution of the product from step B (111 mg, 0.24 mmol, 1 eq.) in 1,4-dioxane (5 ml) was added 2 M aqueous lithium hydroxide (0.24 ml, 0.49 mmol, 2 eq.) and the mixture was heated reflux for 2 hours. The reaction mixture was allowed to cool to ambient temperature and concentrated in vacuo. The crude material was triturated with diethyl ether (10 ml) and the solids were collected by filtration, washed with diethyl ether and dried in vacuo. The resulting material was dissolved in water and acidified to pH 5-6 with 1 M aqueous hydrochloric acid and then concentrated in vacuo. The solids were triturated with 4:1 isopropanol/water (5 mL), collected by filtration and dried in vacuo to give the title product as a yellow solid (66.7 mg, 0.16 mmol, 64%).

HRMS-ERI (m/z) [M+H] ⁺ , calculated for C ₁₉ H ₁₉ N ₆ O ₂ S ₂ : 427.1005; found: 427.1042.

Example 35: 2-({6-[(1,3-benzothiazol-2-yl)amino]-4,5-dimethylpyridazin-3-yl}amino)-5-(1-{[1-(3methoxypropyl)cyclooctyl ]methyl}-5-methyl-1H-pyrazol-4-yl)-1,3-thiazol-4-carboxylic acid

Step A: ethyl 2-[(4,5-dimethyl-6-{[(2Z)-3-{[2-(trimethylsilyl)ethoxy]methyl}-2,3-dihydro-1,3benzothiazol-2-ylidene] amino} pyridazin-3 -yl)amino]-5-(1 -{[1-(3-methoxypropyl)cyclooctyl]methyl}-5methyl-1 H-pyrazol-4-yl)-1,3-thiazol-4- carboxylate.

To a mixture of synthesis product 11a (99 mg, 0.16 mmol, 1 eq.), synthesis product 10b (75.6 mg, 0.19 mmol, 1.2 eq.) and potassium carbonate (64.6 mg, 0 mol, 3 eq.) in a 3:1 mixture tetrahydrofuran/water (4 ml), [1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium(II) (11.4 mg, 0.02 mmol, 0.1 eq.) was added. The mixture was bubbled with nitrogen (10 min) and then heated at 120°C for 2 h under microwave irradiation. The reaction mixture was diluted with ethyl acetate, then washed with water, followed by brine. The organic extract was dried (magnesium sulfate) and the solvent was removed in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 12 g RediSep™ silica gel cartridge) eluting with a gradient of 0-50% ethyl acetate in isoheptane gave the title product as an orange glass (43.4 mg, 0.05 mmol, 33%) .

LC/MS (C ₄₂ H ₆₀ N ₈ O ₄ SiS ₂ ) 834 [M+H] ⁺ ; RT 1.88 (LCMS-V-BI).

1H NMR (400 MHz, DMSOO δ 10.92 (br s, 1H), 7.77 (d, J=7.8 Hz, 1H), 7.56 (s, 1H), 7.45-7.37 (m, 2H),

7.21 (t, J=7.0 Hz, 1H), 5.84 (s, 2H), 4.17 (q, J=7.1 Hz, 2H), 3.85 (s, 2H), 3.71 (t, J=8.0 Hz, 2H), 2.67 (q.3H),

2.41 (s, 3H), 2.36 (s, 3H), 2.33 (q, 2H), 2.19 (s, 3H), 1.65-1.52 (m, 6H), 1.49-1.36 (m, 6H), 1.34-1.22 ( m,8H),

1.18 (t, J=7.1 Hz, 3H), 1.07 (s, 2H), 0.91 (t, 3H), -0.11 (s, 9H).

Step B: ethyl 2-({6-[(1,3-benzothiazol-2-yl)amino]-4,5-dimethylpyridazin-3-yl}amino)-5-(1-{[1-(3methoxypropyl) cyclooctyl]methyl}-5-methyl-1H-pyrazol-4-yl)-1,3-thiazol-4-carboxylate.

A solution of the product from step A (43.4 mg, 0.05 mmol, 1 eq.) in dichloromethane (4 ml) was cooled in an ice bath, then trifluoroacetic acid (0.4 ml, 5.37 mmol, 103 eq.) was added. The mixture was allowed to warm to ambient temperature, at which it was stirred for 18 hours. The reaction mixture was diluted with dichloromethane, then washed successively with a saturated aqueous solution

- 111 046336 sodium bicarbonate, water and saline solution. The organics were dried (magnesium sulfate) and the solvent was removed in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 12 g RediSep™ silica gel cartridge) eluting with a gradient of 0-5% methanol in dichloromethane gave the title product as a yellow glass (21.3 mg, 0.03 mmol, 58%) .

LC/MS (C ₃₆ H ₄₆ N ₈ O ₃ S ₂ ) 703 [M+H]+; RT 1.33 (LCMS-V-Bi).

¹ H NMR (400 MHz, DMSO^6) δ 10.94 (br s, 1H), 7.88 (br s, 1H), 7.63 (br s, 1H), 7.57 (s, 1H), 7.40-7.32 (m, 1H ), 7.22-7.14 (m, 1H), 4.18 (q, J=7.1 Hz, 2H), 3.86 (s, 2H), 3.25 (s, 3H), 2.38 (s + s, 6H), 2.21 (s, 3H), 1.70-1.51 (m, 9H), 1.50-1.35 (m, 6H), 1.34-1.23 (m, 95), 1.18 (t, 3H).

Step C: 2-({6-[(1,3-benzothiazol-2-yl)amino]-4,5-dimethylpyridazin-3-yl}amino)-5-(1-{[1-(3methoxypropyl)cyclooctyl ]methyl}-5-methyl-1H-pyrazol-4-yl)-1,3-thiazol-4-carboxylic acid.

To a solution of the product from step B (21.3 mg, 0.03 mmol, 1 eq.) in 1,4-dioxane (4 ml) was added lithium hydroxide monohydrate (12.7 mg, 0.3 mmol, 10 eq.), and the mixture was refluxed at for 18 hours. The reaction mixture was allowed to cool to ambient temperature and the solvent was removed in vacuo. The residue was triturated with water and the solids were collected by filtration and dried in vacuo to give the title product as a beige solid (13.9 mg, 0.02 mmol, 68%) [as lithium salt].

HRMS-ERI (m/z) [M+H] ⁺ , calculated for C ₃₄ H ₄₃ N ₈ O ₃ S ₂ : 675.2894; found: 675.2933.

Example 36: 2-({6-[(1,3-benzothiazol-2-yl)amino]-4,5-dimethylpyridazin-3-yl}amino)-5-(1-{[1-(3methoxypropyl)cyclooctyl ]methyl}-5-methyl-1H-pyrazol-4-yl)-1,3-thiazol-4-carboxylic acid

Step A: ethyl 2-[(4,5-dimethyl-6-{[(2/)-3-{[3-(trimethylsilyl)propoxy]methyl}-2,3-dihydro-1,3benzothiazol-2-ylidene ]amino}pyridazin-3-yl)amino]-5-(1-{[1-(3-methoxypropyl)cyclohexyl]methyl}-5methyl-1 H-pyrazol-4-yl)-1,3-thiazol-4 -carboxylate.

To a mixture of synthesis product 11a (110 mg, 0.17 mmol, 1 eq.), synthesis product 10c (81 mg, 0.22 mmol, 1.25 eq.) and potassium carbonate (71.4 mg, 0.52 mmol, 3 eq.) in a 3:1 mixture tetrahydrofuran/water (4 ml), [1,G-bis(diphenylphosphino)ferrocene]dichloropalladium(P) (12.6 mg, 0.02 mmol, 0.1 eq.) was added. The mixture was bubbled with nitrogen (10 min) and then heated at 120°C for 1 h under microwave irradiation. The reaction mixture was diluted with ethyl acetate and washed successively with water and brine. The organics were dried (magnesium sulfate) and the solvent was removed in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 12 g RediSep™ silica gel cartridge) eluting with a gradient of 0-50% ethyl acetate in isoheptane gave the title product as an orange gum (81.2 mg, 0.1 mmol, 59%).

LC/MS (C40H56N8O4SiS2) 805 [M+H] ⁺ ; RT 1.85 (LCMS-V-Bi).

1H NMR (400 MHz, DMSO^ ₆ ) δ 10.94 (s, 1H), 7.77 (d, J=7.7 Hz, 1H), 7.56 (s, 1H), 7.47-7.37 (m, 2H), 7.24-7.19 ( m, 1H), 5.84 (s, 2H), 4.17 (q, J=7.1 Hz, 2H), 3.93 (s, 2H), 3.71 (t, 2H), 3.24 (s, 3H), 2.41 (s, 3H ), 2.37 (s, 3H), 2.21 (s, 3H), 1.56-1.47 (m, 5H), 1.42-1.31 (m, 8H), 1.24-1.14 (m, 6H), 0.91 (t, 2H), -0.11 (s, 9Н).

Step B: ethyl 2-({6-[(1,3-benzothiazol-2-yl)amino]-4,5-dimethylpyridazin-3-yl}amino)-5-(1-{[1-(3methoxypropyl) cyclohexyl]methyl}-5-methyl-1H-pyrazol-4-yl)-1,3-thiazol-4-carboxylate.

A solution of the product from Step A (81.2 mg, 0.1 mmol, 1 eq.) in dichloromethane (5 mL) was cooled in an ice bath and trifluoroacetic acid (0.5 mL, 6.71 mmol, 67 eq.) was added. The mixture was allowed to warm to ambient temperature and stirred for 18 hours. The reaction mixture was diluted with dichloromethane, then washed successively with saturated aqueous sodium bicarbonate, water and brine. The organics were dried (magnesium sulfate) and the solvent was removed in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 12 g RediSep™ silica gel cartridge) eluting with a gradient of 0-5% methanol in dichloromethane gave the title product as a yellow glass (49.6 mg, 0.07 mmol, 73%) .

LC/MS (C ₃₄ H ₄₂ N ₈ O ₃ S ₂ ) 675 [M+H] ⁺ ; RT 1.54 (LCMS-V-Bi).

1H NMR (400 MHz, DMSO^ ₆ ) δ 10.94 (br s, 1H), 7.88 (br s, 1H), 7.57 (s, 1H), 7.40-7.32 (m, 1H), 7.227.13 (m, 1H ), 4.18 (q, 2H), 3.95 (s, 2H), 3.25 (s, 3H), 2.37 (s + s, 6H), 2.22 (s, 3H), 1.60-1.45 (m, 6H), 1.44- 1.34 (m, 8H), 1.28-1.22 (m, 2H), 1.19 (t, J=7.1 Hz, 3H).

Step C: 2-({6-[(1,3-benzothiazol-2-yl)amino]-4,5-dimethylpyridazin-3-yl}amino)-5-(1-{[1-(3methoxypropyl)cyclohexyl ]methyl}-5-methyl-1H-pyrazol-4-yl)-1,3-thiazol-4-carboxylic acid.

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To a solution of the product from step B (49.6 mg, 0.07 mmol, 1 eq.) in 1,4-dioxane was added lithium hydroxide monohydrate (14.7 mg, 0.37 mmol, 5 eq.), and the mixture was refluxed for 2 h. The reaction mixture was allowed to cool to ambient temperature and the solvent was removed in vacuo. The residue was triturated with water and the solids were collected by filtration, washed successively with 2 M aqueous hydrochloric acid and then dichloromethane, and dried in vacuo to give the title product as a pale yellow solid (10.8 mg, 0.02 mmol, 23%).

HRMS-ERI (m/z) [M+H]+, calculated for C ₃₂ H ₃₉ N ₈ O ₃ S ₂ : 647.2581; found: 647.2620.

Example 37: 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(ethyl)amino)-1,3-thiazol-4carboxylic acid

Step A: ethyl 2-[(6-chloro-5-methylpyridazin-3-yl)(ethyl)amino]-1,3-thiazol-4-carboxylate.

Sodium hydride (60% dispersion; 150 mg, 3.75 mmol, 1.25 eq.) under nitrogen atmosphere was added to a solution of 3,6-dichloro-4-methylpyridazine (530 mg, 3.25 mmol, 1.09 eq.) and ethyl 2-(ethylamino) -1,3-thiazole-4carboxylate (600 mg, 3 mmol, 1 eq.) in 1,4-dioxane (20 ml) at 0°C. The mixture was stirred for 1 hour at 0°C and for 18 hours at ambient temperature. Ethyl acetate (150 ml) was added and the mixture was washed successively with saturated aqueous ammonium chloride (75 ml), water (2x75 ml) and brine (75 ml), dried (magnesium sulfate) and concentrated in vacuo. Purification by flash column chromatography (50 g silica gel) eluting with dichloromethane gave a pale yellow solid. Trituration with diethyl ether (10 ml) resulted in a solid which was collected by filtration, washed with diethyl ether (2 x 5 ml) and dried in vacuo to give the title product as an off-white solid (475 mg, 1.45 mmol, 49%) .

LC/MS (C ₁₃ H ₁₅ ClN ₄ O ₂ S) 327 [M+H]+; RT 1.25 (LCMS-V-BI).

¹ H NMR (400 MHz, DMSO) δ 8.03 (s, 1H), 7.89 (s, 1H), 4.46 (q, 2H), 4.30 (q, 2H), 2.44 (s, 3H), 1.351.26 (m , 6H).

Step B: ethyl 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(ethyl)amino)-1,3thiazol-4-carboxylate.

2-Aminobenzothiazole (250 mg, 1.66 mmol, 1.2 eq.) and the product from step A (450 mg, 1.38 mmol, 1 eq.) were added to a solution of tris(dibenzylideneacetone)dipalladium(0) (63.1 mg, 0.07 mmol) under a nitrogen atmosphere , 0.05 equiv.) and xantphos (79.7 mg, 0.14 mmol, 0.1 equiv.) in 1,4-dioxane (10 ml). N,Ndiisopropylethylamine (0.75 mL, 4.13 mmol, 3 eq.) was added and the mixture was heated in a sealed tube at 150°C for 7 hours. The solution was allowed to cool to ambient temperature and concentrated in vacuo. Purification by flash column chromatography (50 g silica gel) eluting with ethyl acetate gave a yellow solid. Trituration with methanol (10 ml) resulted in a solid, which was collected by filtration, washed with methanol (2x5 ml) and dried in vacuo to give the title product as a yellow solid (265 mg, 0.6 mmol, 44%).

LC/MS (C20H20N6O2S2) 441 [M+H]+; RT 1.37 (LCMS-V-BI).

1H NMR (400 MHz, DMSO-06) δ 11.12 (br s, 1H), 7.97 (s, 1H), 7.92 (s, 1H), 7.74 (s, 1H), 7.57 (br s, 1H), 7.39 ( t, J=7.5 Hz, 1H), 7.22 (t, J=7.5 Hz, 1H), 4.46 (q, J=6.9 Hz, 2H), 4.30 (q, J=7.1 Hz, 2H), 2.49 (s, 3H), 1.31 (t + t, 6H).

Step C: 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(ethyl)amino)-1,3-thiazol-4carboxylic acid.

To a solution of the product from step B (210 mg, 0.48 mmol, 1 eq.) in a 1:3 mixture of water/1,4-dioxane (4 ml) was added lithium hydroxide monohydrate (80.0 mg, 1.91 mmol, 4 eq.), and the mixture was stirred at ambient temperature overnight. Water (5 ml) was added, the mixture was filtered through celite, and the solids were washed with water (10 ml). The combined filtrate was acidified with acetic acid to give a yellow slurry, and the solids were collected by filtration, washed with water (30 mL) and dried in vacuo to give the title product as a yellow solid (170 mg, 0.41 mmol, 86%).

HRMS-ERI (m/z) [M+H]+, calculated for C ₁₈ H ₁₇ N ₆ O ₂ S ₂ : 413.0849; found: 413.0882.

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Example 38: 6-({6-[(1,3-Benzothiazol-2-yl)amino]pyridazin-3-yl} (methyl)amino)pyridine-2-carboxylic acid

Step A: tert-butyl N[(tert-butoxy)carbonyl]-N-(6-chloropyridazin-3-yl)carbamate.

4-Dimethylaminopyridine (122 mg, 1 mmol, 0.02 eq.) was added under nitrogen atmosphere to a suspension of 3-amino-6-chloropyridazine (6.5 g, 50.2 mmol, 1 eq.) and di-tert-butyl dicarbonate (25.2 g, 115 mmol, 2.3 eq.) in dimethylformamide (45 ml). The suspension was heated at 50°C for 3 hours, then allowed to cool to ambient temperature and water (350 ml) was added. The solids were collected by filtration, washed with water (250 ml) then dissolved in dichloromethane (250 ml) and the solution filtered through silica gel (25 g) eluting with dichloromethane. The filtrate was concentrated in vacuo and triturated with heptane (50 ml) to give a solid, which was collected by filtration, washed with heptane (50 ml) and dried in vacuo to give the title product as a white solid (9.85 g, 29.9 mmol, 60% ).

LC/MS (C ₁₄ H ₂₀ ClN ₃ O ₄ ) 174 [M-Boc-tBu+3H]+; RT 1.28 (LCMS-V-BI).

¹ H NMR (400 MHz, DMSO-de) δ 7.53 (d, 2H), 1.46 (s, 18H).

Step B: methyl 6-[(6-{bis[(tert-butoxy)carbonyl]amino}pyridazin-3-yl)amino]pyridine-2carboxylate.

Methyl 6-aminopyridine-2-carboxylate (1.65 g, 10.8 mmol, 1.1 eq.) and the product from step A (3.25 g, 9.86 mmol, 1 eq.) were added to a solution of tris(dibenzylideneacetone)dipalladium(0) ( 451 mg, 0.49 mmol, 0.05 eq.) and xantphos (570 mg, 0.99 mmol, 0.1 eq.) in 1,4-dioxane (60 ml). Na-diisopropylethylamine (5.2 mL, 29.6 mmol, 3 eq.) was added and the mixture was heated at 100°C for 3 hours. The solution was allowed to cool to ambient temperature and concentrated in vacuo. The residue was partitioned between dichloromethane (300 ml) and water (200 ml), and the organic phase was washed successively with water (200 ml) and brine (150 ml), then dried (magnesium sulfate) and concentrated in vacuo. Purification by flash column chromatography (100 g silica gel) eluting with 2:3 ethyl acetate/heptane gave a yellow solid. Trituration with diethyl ether (20 ml) provided a solid, which was collected by filtration, washed with diethyl ether (20 ml) and dried in vacuo to give the title product as a pale yellow solid (3.35 g, 7.52 mmol, 76%) .

LC/MS (C ₂₁ H ₂ 7N ₅ O ₆ ) 446 [M+H] ⁺ ; RT 1.27 (LCMS-V-BI).

1H NMR (400 MHz, DMSO-do) δ 10.41(s, 1H), 9.02 (d, 1H), 7.97 (dd, 1H), 7.83-7.73 (m, 2H), 7.45 (d, 1H), 4.01 ( s, 3H), 1.46 (s, 9H).

Step C: methyl 6-[(6-{bis[(tert-butoxy)carbonyl]amino}pyridazin-3-yl)(methyl)amino]pyridine 2-carboxylate.

Potassium tert-Butoxide (750 mg, 6.68 mmol, 1.32 eq.) was added under nitrogen to a suspension of the product from Step B (2.25 g, 5.05 mmol, 1 eq.) in dimethyl sulfoxide (25 ml) and the mixture was stirred for 30 min. . Methyl iodide (0.5 mL, 8.03 mmol, 1.59 eq.) was added and the suspension was stirred for 3 hours. The mixture was partitioned between ethyl acetate (250 mL) and saturated aqueous ammonium chloride (150 mL), washed with water (2x150 mL) and brine (100 ml), dried (magnesium sulfate) and concentrated in vacuo. Purification by flash column chromatography (100 g silica gel) eluting with 2:3 ethyl acetate/heptane provided the title product as a yellow solid (1.4 g, 2.59 mmol, 51%), which was used directly in the next step without further cleaning.

LC/MS (C ₂₂ H ₂ 9N ₅ O ₆ ) 460 [M+H] ⁺ ; RT 1.29 (LCMS-V-BI).

Step D: Methyl 6-[(6-aminopyridazin-3-yl)(methyl)amino]pyridine-2-carboxylate.

Trifluoroacetic acid (5 ml, 64.7 mmol, 25 eq.) at 0°C was added to the product from step C (1.4 g, 2.59 mmol, 1 eq.) in dichloromethane (35 ml), and the solution was stirred for 1 h at 0 °C and then for 18 hours at ambient temperature. The solution was concentrated in vacuo and water (25 ml) was added to give a pale yellow suspension, which was neutralized with sodium bicarbonate and the solids were collected by filtration, washed with water (75 ml) and dried in vacuo to give the title product as a yellow solid (650 mg, 2.51 mmol, 97%), which was directly used in the next step without further purification or characterization.

Step E: methyl 6-({6-[(1,3-benzothiazol-2-yl)amino]pyridazin-3-yl}(methyl)amino)pyridine-2-carboxylate.

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The product from step D (650 mg, 2.51 mmol, 1 eq.) and 2-bromo-1,3-benzothiazole (600 mg, 2.8 mmol, 1.12 eq.) were added to the tris(dibenzylideneacetone)dipalladium(0) solution under nitrogen atmosphere. (115 mg, 0.13 mmol, 0.05 eq.) and xantphos (145 mg, 0.25 mmol, 0.1 eq.) in 1,4-dioxane (10 ml). N,Ndiisopropylethylamine (1.4 mL, 7.52 mmol, 3 eq.) was added and the mixture was heated at 100°C for 24 hours. The reaction mixture was allowed to cool to ambient temperature and concentrated in vacuo. Water (20 ml) was added to give a brown slurry and the solids were collected by filtration, washed with water (40 ml) and dried in vacuo. Purification by flash column chromatography (50 g silica gel) eluting with 39:1 dichloromethane/methanol gave a solid which was triturated with methanol (10 ml), filtered, washed with methanol (10 ml) and dried in vacuo to give the title product in as a pale brown solid (225 mg, 0.57 mmol, 23%).

LC/MS (C ₁₉ H ₁₆ N ₆ O ₂ S) 393 [M+H]+; RT 1.14 (LCMS-V-BI).

1H NMR (400 MHz, DMSO-de) δ 11.73 (br s, 1H), 7.94 (d, 1H), 7.91-7.85 (m, 1H), 7.80 (d, J=9.5 Hz, 1H), 7.70-7.59 (m, 2H), 7.46 (d, 1H), 7.43-7.35 (m, 2H), 7.28-7.19 (m, 1H), 3.86 (s, 3H), 3.64 (s, 3H).

Step F: 6-({6-[(1,3-benzothiazol-2-yl)amino]pyridazin-3-yl}(methyl)amino)pyridine-2-carboxylic acid.

To a solution of the product from step E (200 mg, 0.51 mmol, 1 eq.) in 1:3 water/1,4-dioxane (4 mL) was added lithium hydroxide monohydrate (85.5 mg, 2.04 mmol, 4 eq.), and the mixture was stirred at ambient temperature for 2 hours. Water (5 ml) was added, the solution was filtered through celite, and the solids were washed with water (10 ml). The combined filtrate was acidified with acetic acid to give a solid, which was collected by filtration, washed with water (20 mL) and dried in vacuo to give the title product as a yellow solid (160 mg, 0.42 mmol, 83%).

MSVR-ERI (m/z) [M+H]+, calculated for С^^ОзЗ: 379.0972; found: 379.0993.

Example 39: 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl} (2-methoxyethyl)amino)1,3-thiazol-4-carboxylic acid

Step A: ethyl 2-[(6-chloro-5-methylpyridazin-3-yl)(2-methoxyethyl)amino]-1,3-thiazol-4carboxylate.

Sodium hydride (60% dispersion; 650 mg, 16.3 mmol, 1.2 eq.) under nitrogen atmosphere was added to a solution of synthesis product 3z (3.15 g, 13.7 mmol, 1 eq.) and 3,6-dichloro-4-methylpyridazine (2.5 g , 15.3 mmol, 1.12 eq.) in 1,4-dioxane (50 ml) at 0°C. The mixture was stirred for 1 hour at 0°C, then heated at 60°C for 24 hours. The mixture was allowed to cool to ambient temperature, then partitioned between ethyl acetate (300 ml) and saturated aqueous ammonium chloride (150 ml), and the organic phase was washed successively with water (2x150 ml) and brine (100 ml), dried (magnesium sulfate) and concentrated in vacuo. Trituration with diethyl ether (25 mL) provided a solid, which was collected by filtration, washed with diethyl ether (15 mL) and dried in vacuo to give the title product as a pale pink solid (2.55 g, 7.15 mmol, 52%) .

LC/MS (C ₁₄ H ₁₇ ClN ₄ O3S) 357 [M+H]+; RT 1.22 (LCMS-V-BI).

1H NMR (400 MHz, CDCI3) δ 7.83 (s, 1H), 7.65 (s, 1H), 4.60 (t, 2H), 4.39 (q, 2H), 3.91 (t, 2H), 3.32 (s, 3H) , 2.46 (s, 3H), 1.41 (t, 3H).

Stage

IN:

ethyl

2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(2 methoxyethyl)amino)-1,3-thiazol-4-carboxylate.

2-Aminobenzothiazole (1.15 g, 7.66 mmol, 1.09 eq.) and the product from step A (2.5 g, 7.01 mmol, 1 eq.) were added to a solution of tris(dibenzylideneacetone)dipalladium(0) (321 mg, 0.35 mmol) under a nitrogen atmosphere , 0.05 eq.) and xantphos (405 mg, 0.7 mmol, 0.1 eq.) in 1,4-dioxane (45 ml). N,Ndiisopropylethylamine (3.8 mL, 21.0 mmol, 3 eq.) was added and the mixture was heated in a stoppered flask at 150°C for 18 hours. The solution was allowed to cool to ambient temperature, filtered through celite, and the solids were washed with dichloromethane. The combined filtrate was concentrated in vacuo and triturated with methanol (25 ml) to give a solid, which was collected by filtration, washed with methanol (3x15 ml) and dried in vacuo to give the title product as an orange solid (2.55 g, 5.42 mmol, 77 %).

LC/MS (C ₂₁ H ₂₂ N ₆ O3S ₂ ) 471 [M+H]+; RT 1.35 (LCMS-VB!).

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1H NMR (400 MHz, CDCl ₃ ) δ 7.85-7.80 (m, 1H), 7.79 (s, 1H), 7.77-7.69 (m, 1H), 7.66 (d, J=1.3 Hz, 1H), 7.48-7.36 (m, 1H), 7.31-7.21 (m, 1H), 4.64-4.48 (m, 2H), 4.41 (q, J=7.1 Hz, 2H), 3.94-3.85 (m, 2H), 3.40-3.31 (m , 3H), 2.44 (s, 3H), 1.43 (t, J=7.1 Hz, 3H).

Step C: 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(2-methoxyethyl)amino)-1,3thiazol-4-carboxylic acid.

To a solution of the product from step B (250 mg, 0.53 mmol, 1 eq.) in 1:3 water/1,4-dioxane (4 ml) was added lithium hydroxide monohydrate (89.2 mg, 2.13 mmol, 4 eq.), and the mixture was stirred at ambient temperature for 18 hours. Water (10 ml) was added and the suspension was acidified with acetic acid. The solids were collected by filtration, washed with water (3 x 25 mL) and dried in vacuo to give the title product as a yellow solid (175 mg, 0.4 mmol, 74%).

HRMS-ERI (m/z) [M+H]+, calculated for C ₁₉ H ₁₉ N ₆ O ₃ S ₂ : 443.0955; found: 443.0983.

Example 40: 5-{1-[(adamantan-1-yl)methyl]-5-methyl-1H-pyrazol-4-yl}-2-({6-[(1,3-benzothiazol-2yl)amino] -5-methylpyridazin-3-yl}(methyl)amino)-1,3-thiazol-4-carboxylic acid

Step A: ethyl 5-{1-[(adamantan-1-yl)methyl]-5-methyl-1H-pyrazol-4-yl}-2-[methyl(5-methyl-6-{[(2Z)3 -{[2-(trimethylsilyl)ethoxy]methyl}-2,3-dihydro-1,3-benzothiazol-2-ylidene]amino}pyridazin-3yl)amino]-1,3-thiazol-4-carboxylate.

To a mixture of synthesis product 11b (54.7 mg, 0.09 mmol, 1 eq.), synthesis product 10a (36.8 mg, 0.1 mmol, 1.2 eq.) and potassium carbonate (35.7 mg, 0.26 mmol, 3 eq.) in a 3:1 mixture tetrahydrofuran/water (4 ml), [1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium(II) (6.3 mg, 0.01 mmol, 0.1 eq.) was added. The mixture was bubbled with nitrogen (10 min) and then heated at 120°C for 1 h under microwave irradiation. The reaction mixture was partitioned between ethyl acetate and water, and the organic phase was washed with brine, dried (magnesium sulfate) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 12 g RediSep™ silica gel cartridge) eluting with a gradient of 0-50% ethyl acetate in isoheptane gave the title product as a brown glass (46.7 mg, 0.06 mmol, 69%) .

LC/MS (C40H52N8O ₃ SiS2) 785 [M+H] ⁺ ; RT 1.91 (LCMS-V-BI).

1H NMR (400 MHz, DMSO^) δ 7.82 (dd, J=7.7, 1.1 Hz, 1H), 7.71 (d, J=1.1 Hz, 1H), 7.53 (s, 1H), 7.487.39 (m, 2H ), 7.23 (ddd, J=8.3, 7.0, 1.5 Hz, 1H), 5.86 (s, 2H), 4.16 (q, J=7.1 Hz, 2H), 3.82 (s, 3H), 3.78 (s, 2H) , 3.71 (t, 2H), 2.46 (s, 3H), 2.18 (s, 3H), 1.95 (s, 3H), 1.72-1.54 (m, 12H), 1.16 (t, J=7.1 Hz, 3H), 0.92 (t, 3H), 0.11 (s, 9H).

Step B: ethyl 5-{1-[(adamantan-1-yl)methyl]-5-methyl-1H-pyrazol-4-yl}-2-({6-[(1,3-benzothiazol-2yl)amino ]-5-methylpyridazin-3-yl}(methyl)amino)-1,3-thiazol-4-carboxylate.

A solution of the product from step A (46.7 mg, 0.06 mmol, 1 eq.) in dichloromethane (3 ml) was cooled in an ice bath, then trifluoroacetic acid (0.3 ml, 4.03 mmol, 67.7 eq.) was added and the mixture was allowed to warm to ambient temperature and stirred for 18 hours. The reaction mixture was partitioned between dichloromethane and saturated aqueous sodium bicarbonate, and the organic phase was washed successively with water and brine, then dried (magnesium sulfate) and the solvent was removed in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 12 g RediSep™ silica gel cartridge) eluting with a gradient of 0-5% methanol in dichloromethane gave the title product as a yellow glass (24.2 mg, 0.04 mmol, 62%) .

LC/MS (C ₃₄ H ₃₈ N ₈ O ₂ S ₂ ) 655 [M+H] ⁺ ; RT 1.53 (LCMS^-B2).

1H NMR (400 MHz, DMSO-do) δ 7.90 (br s, 1H), 7.73 (s, 1H), 7.54 (s, 1H), 7.37 (t, J=7.7 Hz, 1H), 7.247.15 (m , 1H), 4.17 (q, J=7.1 Hz, 2H), 3.81 (s, 3H), 3.79 (s, 2H), 2.47 (s, 3H), 2.20 (s, 3H), 1.97 (s, 3H) , 1.72-1.55 (m, 12H), 1.17 (t, J=7.1 Hz, 3H).

Step C: 5-{1-[(adamantan-1-yl)methyl]-5-methyl-1H-pyrazol-4-yl}-2-({6-[(1,3-benzothiazol-2yl)amino ]-5-methylpyridazin-3-yl}(methyl)amino)-1,3-thiazol-4-carboxylic acid.

To a solution of the product from step B (24.2 mg, 0.04 mmol, 1 eq.) in 1,4-dioxane (3 ml) was added lithium hydroxide monohydrate (7.8 mg, 0.18 mmol, 5 eq.), and the mixture was refluxed at for 18 hours. The reaction mixture was allowed to cool to ambient temperature, then the solvent was removed in vacuo. The residue was triturated with water, filtered and dried in vacuo to give the title product as a yellow solid (16.3 mg, 0.03 mmol, 70%) [as lithium salt].

HRMS-ERI (m/z) [M+H] ⁺ , calculated for C ₃₂ H ₃₅ N ₈ O ₂ S ₂ : 627.2319; found: 627.2358.

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Example 41: 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}[2-(morpholin-4yl)ethyl]amino)-1,3-thiazole -4-carboxylic acid

Step A: ethyl 2-[(2,2-dimethoxyethyl)amino]-1,3-thiazole-4-carboxylate.

Ammonia (2 M in isopropanol; 25 mL, 50 mmol, 2.1 eq.) under nitrogen atmosphere was added to a solution of 1,1-dimethoxy-2-isothiocyanatoethane (3.5 g, 20 mmol, 1 eq.) in isopropanol (15 mL) at 0°C, and the mixture was stirred for 1 hour at 0°C and then for 18 hours at ambient temperature. The solvent was removed in vacuo and the residue was dissolved in ethanol (60 ml) and cooled to 0°C. Ethyl bromopyruvate (3.6 mL, 25.8 mmol, 1.09 eq.) was added to the solution under nitrogen atmosphere and the mixture was allowed to warm to ambient temperature and stirred for 1.5 h. Triethylamine (5.6 mL, 39.9 mmol, 2 eq.) was added and the mixture was stirred. for 2 hours and then concentrated in vacuo. The residue was partitioned between ethyl acetate (250 ml) and water (150 ml), and the organic phase was washed successively with water (150 ml) and brine (150 ml), dried (magnesium sulfate) and concentrated in vacuo. Purification by flash column chromatography (100 g silica gel) eluting with 2:3 ethyl acetate/heptane gave a pale yellow solid, which was triturated with heptane (40 ml), filtered, washed with heptane (40 ml) and dried in vacuum to obtain the title product as an off-white solid (2.95 g, 11.33 mmol, 48%).

LC/MS (C ₁₀ H ₁₆ N ₂ O ₄ S) 261 [M+H]+; RT 0.92 (LCMS-V-BI).

¹ H NMR (400 MHz, CDCl ₃ ) δ 7.41 (s, 1H), 5.59 (t, J=5.5 Hz, 1H), 4.52 (t, J=5.4 Hz, 1H), 4.35 (q, J=7.1 Hz , 2H), 3.41 (dd + s, 8H), 1.36 (t, J=7.1 Hz, 3H).

Step B: ethyl 2-[(6-chloro-5-methylpyridazin-3-yl)(2,2-dimethoxyethyl)amino]-1,3-thiazol-4carboxylate.

Sodium hydride (60% dispersion; 500 mg, 12.5 mmol, 1.25 eq.) under nitrogen was added to a solution of the product from step A (2.6 g, 9.99 mmol, 1 eq.) and 3,6-dichloro-4-methylpyridazine (1.85 g, 11.4 mmol, 1.14 eq.) in 1,4-dioxane (40 ml) at 0°C, and the mixture was stirred for 1 hour at 0°C and then for 5 hours at 60°C. The solution was allowed to cool to ambient temperature, then partitioned between ethyl acetate (300 ml) and saturated aqueous ammonium chloride (150 ml). The organic phase was washed successively with water (2x150 ml) and brine (150 ml), dried (magnesium sulfate) and concentrated in vacuo. Purification by flash column chromatography (100 g silica gel) eluting with 99:1 dichloromethane/methanol gave an orange gum which was triturated with diethyl ether (15 ml) to give a solid which was collected by filtration, washed with diethyl ether ( 20 ml) and dried in vacuum to obtain the target product in the form of a pale pink powder (1.95 g, 5.04 mmol, 50%).

LC/MS (C ₁₅ H ₁₉ ClN ₄ O ₄ S) 387 [M+H] ⁺ ; RT 1.26 (LCMS-V-BI).

1H NMR (400 MHz, DMSO-d,) δ 8.06 (s, 1H), 7.95 (d, J=1.1 Hz, 1H), 4.78 (t, J=5.2 Hz, 1H), 4.51 (d, J=5.2 Hz, 2H), 4.29 (q, J=7.1 Hz, 2H), 3.34 (s, 6H), 2.42 (d, J=0.9 Hz, 3H), 1.31 (t, J=7.1 Hz, 3H).

Step C: Ethyl 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(2,2dimethoxyethyl)amino)-1,3-thiazol-4-carboxylate .

The product from step B (1.6 g, 4.14 mmol, 1 eq.) and 2-aminobenzothiazole (700 mg, 4.66 mmol, 1.13 eq.) were added to a solution of tris(dibenzylideneacetone)dipalladium(0) (189 mg, 0.21 mmol) under a nitrogen atmosphere , 0.05 eq.) and xantphos (239 mg, 0.41 mmol, 0.1 eq.) in 1,4-dioxane (20 ml), and the mixture was stirred. Diisopropylethylamine (2.2 mL, 12.4 mmol, 3 eq.) was added and the mixture was heated in a sealed tube at 150°C for 18 hours. The solution was allowed to cool to ambient temperature and concentrated in vacuo. Trituration with diethyl ether (20 ml) provided a solid which was collected by filtration, washed with diethyl ether (2 x 10 ml) and dried in vacuo to give the title product as a yellow/green solid (1.9 g, 3.42 mmol, 83%) , which was used in the next step without further purification.

LC/MS (C ₂₂ H ₂₄ N ₆ O ₄ S ₂ ) 501 [M+H] ⁺ ; RT 1.37 (LCMS-V-BI).

1H NMR (400 MHz, DMSO) δ 11.11 (br s, 1H), 7.99 (s, 1H), 7.92 (br s, 1H), 7.78 (s, 1H), 7.57 (br s, 2H), 7.40 (t , J=7.6 Hz, 1H), 7.22 (t, J=7.6 Hz, 1H), 4.78 (t, J=5.2 Hz, 1H), 4.48 (d, J=5.2 Hz, 2H), 4.29 (q, J =7.1 Hz, 2H), 3.35 (s, 6H), 2.46 (s, 3H), 1.31 (t, J=7.1 Hz, 3H).

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Step D: ethyl 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}[2-(morpholin-4yl)ethyl]amino)-1,3- thiazole-4-carboxylate.

Formic acid (10 mL, 0.27 mol, 177 eq.) was added to a suspension of the product from step C (750 mg, 1.5 mmol, 1 eq.) in water (5 mL) and the mixture was heated at 85°C for 3 h. The solution was allowed to cool to ambient temperature and concentrated in vacuo. Dichloromethane (20 mL) was added followed by morpholine (0.5 mL, 5.78 mmol, 3.86 eq.) and sodium triacetoxyborohydride (1.59 g, 7.49 mmol, 5 eq.) and the mixture was stirred for 18 h. The reaction mixture was partitioned between dichloromethane ( 125 ml) and 10% aqueous potassium carbonate (75 ml), and the organic phase was washed successively with water (2x75 ml) and brine (75 ml), dried (magnesium sulfate) and concentrated in vacuo. Purification by flash column chromatography (20 g silica gel) eluting with 19:1 dichloromethane/methanol gave a yellow solid, which was triturated with diethyl ether (15 ml), filtered, washed with diethyl ether (15 ml), and dried in vacuo to give the title product as a pale orange solid (375 mg, 0.71 mmol, 48%).

LC/MS (C ₂₄ H ₂₇ N ₇ O ₃ S ₂ ) 526 [M+H]+; RT 1.08 (LCMS-V-BI).

1H NMR (400 MHz, DMSO^) δ 11.18 (br s, 1H), 7.97 (s, 1H), 7.91 (br s, 1H), 7.75 (s, 1H), 7.55 (br s, 1H), 7.39 ( t, 1H), 7.22 (t, J=7.6 Hz, 1H), 4.53 (t, J=6.5 Hz, 2H), 4.29 (q, J=7.1 Hz, 2H), 3.48 (t, J=4.5 Hz, 4H), 2.65 (t, J=6.5 Hz, 2H), 2.47 (s, 3H), 1.31 (t, J=7.1 Hz, 3H).

Step E: 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}[2-(morpholin-4yl)ethyl]amino)-1,3-thiazole -4-carboxylic acid.

To a solution of the product from step D (275 mg, 0.52 mmol, 1 eq.) in 2:3 water/1,4-dioxane (5 mL) was added lithium hydroxide monohydrate (87.8 mg, 2.09 mmol, 4 eq.), and the mixture was stirred at ambient temperature for 18 hours. Water (10 ml) was added and the mixture was acidified with acetic acid. The solids were collected by filtration, washed with water and dried in vacuo to give the title product as a pale orange solid (215 mg, 0.43 mmol, 83%).

MSVR-ERI (m/z) [MH] ^- , calculated for C ₂₂ H ₂₂ N ₇ O ₃ S ₂ : 496.1231; found: 496.1232.

Example 42: 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)-5-(1-{[1-(3methoxypropyl) cyclohexyl]methyl}-5-methyl-1H-pyrazol-4-yl)-1,3-thiazol-4-carboxylic acid

Step A: ethyl 5-(1-{[1-(3-methoxypropyl)cyclohexyl]methyl}-5-methyl-1H-pyrazol-4-yl)-2[methyl(5-methyl-6-{[(2Z )-3-{[2-(trimethylsilyl)ethoxy]methyl}-2,3-dihydro-1,3-benzothiazol-2ylidene] amino } pyridazin-3-yl)amino]-1,3-thiazol-4-carboxylate .

To a mixture of synthesis product 11b (110 mg, 0.17 mmol, 1 eq.), synthesis product 10c (78.2 mg, 0.21 mmol, 1.2 eq.) and potassium carbonate (71.8 mg, 0.52 mmol, 3 eq.) in a 10:1 mixture tetrahydrofuran (4 ml)/water (0.4 ml) [1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium(II) (12.7 mg, 0.02 mmol, 0.1 eq.) was added. The mixture was bubbled with nitrogen (10 min) and then heated at 120°C for 1 h under microwave irradiation. The reaction mixture was partitioned between ethyl acetate and water, and the organic phase was washed with brine, dried (magnesium sulfate), and the solvent was removed in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 12 g RediSep™ silica gel cartridge) eluting with a gradient of 0-50% ethyl acetate in isoheptane gave the title product as a yellow foam (56.1 mg, 0.07 mmol, 40%).

LC/MS (C40H56N8O4SiS2) 805 [M+H] ⁺ ; RT 1.74 (ZHMS-U-V2).

1H NMR (400 MHz, DMSO^) δ 7.92 (dd, J=7.6, 1.0 Hz, 1H), 7.82 (d, J=1.1 Hz, 1H), 7.66 (s, 1H), 7.617.47 (m, 2H ), 7.35 (ddd, J=8.3, 7.0, 1.5 Hz, 1H), 5.96 (d, J=9.8 Hz, 2H), 4.28 (q, J=7.1 Hz, 2H), 4.05 (s, 2H), 3.93 (s, 3H), 3.83 (t, 2H), 3.35 (s, 3H), 2.58 (d, J=1.0 Hz, 3H), 2.31 (s, 3H), 1.69-1.58 (m, 5H), 1.52- 1.44 (m, 7H), 1.38-1.32 (s, 4H), 1.28 (t, J=7.1 Hz, 3H), 1.03 (t, 2H), 0.00 (s, 9H).

Step B: ethyl 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)-5-(1{[ 1 -(3- methoxypropyl)cyclohexyl]methyl }-5 -methyl-1 H-pyrazol-4-yl)-1, 3 -thiazol-4-carboxylate.

A solution of the product from Step A (56.1 mg, 0.07 mmol, 1 eq.) in dichloromethane (3 mL) was cooled in an ice bath and trifluoroacetic acid (0.5 mL, 6.71 mmol, 96 eq.) was added. The mixture was allowed to warm to ambient temperature, at which it was stirred overnight. The solvent was removed in vacuo and the crude product was purified using reverse phase automated flash chromatography (CombiFlash Rf, C18 13 g RediSep column) eluting with a 5-95% aceto gradient

- 118 046336 nitrile (pH 4; formic acid) in water (pH 4; formic acid) to obtain the target product in the form of a yellow glassy substance (34.2 mg, 0.05 mmol, 73%).

LC/MS (C ₃₄ H ₄₂ N ₈ O ₃ S ₂ ) 675 [M+H]+; RT 1.47 (LCMS^-B2).

¹ H NMR (400 MHz, DMSO-06) δ 7.88 (br s, 1H), 7.73 (s, 1H), 7.55 (s, 1H), 7.52 (br s, 1H), 7.37 (t, 1H), 7.19 (t, 1H), 4.17 (q, J=7.1 Hz, 2H), 3.94 (s, 2H), 3.82 (s, 3H), 3.24 (s, 3H), 2.48 (s, 3H), 2.20 (s, 3H), 1.60-1.45 (m, 6H), 1.45-1.31 (m, 8H), 1.29-1.21 (m, 2H), 1.17 (t, 3H).

Step C: 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)-5-(1-{[1-(3methoxypropyl) cyclohexyl]methyl}-5-methyl-1H-pyrazol-4-yl)-1,3-thiazol-4-carboxylic acid.

To a solution of the product from step B (34 mg, 0.05 mmol, 1 eq.) in 1,4-dioxane (3 ml) was added lithium hydroxide monohydrate (10.6 mg, 0.25 mmol, 5 eq.), and the mixture was refluxed at during the night. The reaction mixture was allowed to cool to ambient temperature, then the solvent was removed in vacuo. The residue was diluted with water and acidified with 2 N. aqueous hydrochloric acid, and extracted with a 3:1 mixture of dichloromethane/isopropanol. The organic extracts were dried (magnesium sulfate), concentrated in vacuo, and dried in vacuo to give the title product as a cream solid (16.7 mg, 0.03 mmol, 51%).

HRMS-ERI (m/z) [M+H] ⁺ , calculated for C ₃₂ H ₃₉ N ₈ O ₃ S ₂ : 647.2581; found: 647.2591.

Example 43: 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)-5-(1-{[1-(3methoxypropyl) cyclooctyl]methyl}-5-methyl-1H-pyrazol-4-yl)-1,3-thiazol-4-carboxylic acid

Step A: ethyl 5-(1-{[1-(3-methoxypropyl)cyclooctyl]methyl}-5-methyl-1H-pyrazol-4-yl)-2[methyl(5-methyl-6-{[(2Z )-3-{[2-(trimethylsilyl)ethoxy]methyl}-2,3-dihydro-1,3-benzothiazol-2ylidene] amino } pyridazin-3-yl)amino]-1,3-thiazol-4-carboxylate .

To a mixture of synthesis product 11b (106 mg, 0.17 mmol, 1 eq.), synthesis product 10b (80.9 mg, 0.2 mmol, 1.2 eq.) and potassium carbonate (69.1 mg, 0.5 mmol, 3 eq.) in a 3:1 mixture tetrahydrofuran/water (4 ml), [1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium(II) (12.2 mg, 0.02 mmol, 0.1 eq.) was added. The mixture was bubbled with nitrogen (10 min) and then heated at 120°C for 1 h under microwave irradiation. The reaction mixture was partitioned between ethyl acetate and water, and the organic phase was washed with brine, dried (magnesium sulfate), and the solvent was removed in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 12 g RediSep™ silica gel cartridge) eluting with a gradient of 0-50% ethyl acetate in isoheptane gave the title product as a colorless gum (81.3 mg, 0.1 mmol, 59%).

LC/MS (C42H60N8O4SiS2) 833 [M+H] ⁺ ; RT 1.81 (LCMS^-B2).

1H NMR (400 MHz, DMSO-06) δ 7.92 (d, J=7.6 Hz, 1H), 7.82 (s, 1H), 7.66 (s, 1H), 7.61-7.50 (m, 2H), 7.37-7.28 ( m, 1H), 5.98 (s, 2H), 4.28 (q, J=7.1 Hz, 2H), 3.96 (s, 2H), 3.93 (s, 3H), 3.83 (t, J=8.0 Hz, 2H), 3.35 (s, 3H), 2.58 (s, 3H), 2.29 (s, 3H), 1.76-1.64 (m, 9H), 1.60-1.45 (m, 5H), 1.46-1.33 (m, 6H), 1.28 ( t, J=7.1 Hz, 3H), 1.03 (t, 3H), 0.00 (s, 9H).

Step B: ethyl 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)-5-(1{[1-(3- methoxypropyl)cyclooctyl]methyl}-5-methyl-1H-pyrazol-4-yl)-1,3-thiazol-4-carboxylate.

A solution of the product from step A (81.3 mg, 0.1 mmol, 1 eq.) in dichloromethane (3 ml) was cooled in an ice bath, then trifluoroacetic acid (0.3 ml, 4.03 mmol, 41 eq.) was added and the mixture was stirred at ambient temperature over 18 hours. The solvent was removed in vacuo and the crude product was purified by automated flash column chromatography (CombiFlash Rf, 12 g RediSep™ silica gel cartridge) eluting with a gradient of 0-5% methanol in dichloromethane to give the title product as a white solid substances (81.5 mg, 0.12 mmol, > 100%).

LC/MS (C ₃₆ H ₄₆ N ₈ O ₃ S ₂ ) 703 [M+H] ⁺ ; RT 1.56 (LCMS^-B2).

1H NMR (400 MHz, DMSO-06) 7.88 (br s, 1H), 7.73 (s, 1H), 7.56 (s, 1H), 7.50 (br s, 1H), 7.38 (t, J=7.5 Hz, 1H ), 7.20 (t, J=7.6 Hz, 1H), 4.17 (q, J=7.1 Hz, 2H), 3.85 (s, 2H), 3.81 (s, 3H), 3.33 (t, J=6.5 Hz, 2H ), 3.25 (s, 3H), 2.48 (s, 3H), 2.19 (s, 3H), 1.66-1.54 (m, 9H), 1.49-1.36 (m, 4H), 1.33-1.22 (m, 5H), 1.18 (t, J=7.1 Hz, 3H).

Step C: 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)-5-(1-{[1-(3methoxypropyl) cyclooctyl]methyl}-5-methyl-1H-pyrazol-4-yl)-1,3-thiazol-4-carboxylic acid.

To a solution of the product from step B (81.5 mg, 0.12 mmol, 1 eq.) in 1,4-dioxane (5 ml) was added lithium hydroxide monohydrate (24.3 mg, 0.58 mmol, 5 eq.), and the mixture was refluxed at during the night. The reaction mixture was allowed to cool to ambient temperature and the solvent

- 119 046336 was removed in vacuum. The residue was diluted with water, acidified with 2N. aqueous hydrochloric acid, and extracted with a 3:1 mixture of dichloromethane/isopropanol. The organic extracts were dried (magnesium sulfate) and concentrated under reduced pressure to give the title product as a cream solid (53.6 mg, 0.08 mmol, 69%).

HRMS-ERI (m/z) [M+H]+, calculated for C ₃₄ H ₄₃ N ₈ O ₃ S ₂ : 675.2894; found: 675.2890.

Example 44: 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}[3-(morpholin-4yl)propyl]amino)-1,3-thiazole -4-carboxylic acid

Step A: ethyl 2-{[3-(morpholin-4-yl)propyl]amino}-1,3-thiazol-4-carboxylate.

Ethyl bromopyruvate (2.5 mL, 17.9 mmol, 1.18 eq.) was added under nitrogen to a suspension of 3(morpholin-4-yl)propylthiourea (3.1 g, 15.3 mmol, 1 eq.) in ethanol (50 mL) at 0°C, and the mixture was stirred for 1 hour at ambient temperature. Triethylamine (4.5 mL, 30.5 mmol, 2 eq.) was added and the mixture was heated at 85°C for 4 hours. The solution was allowed to cool to ambient temperature and concentrated in vacuo. 10% aqueous potassium carbonate (150 ml) was added and the mixture was extracted with ethyl acetate (5x100 ml). The combined organic extracts were washed with water (2x150 ml) and brine (100 ml), then dried (magnesium sulfate) and concentrated in vacuo. Purification by flash column chromatography (100 g silica gel) eluting with 19:1 dichloromethane/methanol (19:1) gave a solid which was triturated with heptane (50 ml), filtered, washed with heptane (50 ml) and dried in vacuo to give the title product as a pale brown solid (4 g, 13.4 mmol, 88%).

LC/MS (C ₁₃ H ₂₁ N ₃ O ₃ S) 300 [M+H] ⁺ ; RT 0.58 (LCMS-V-BI).

¹ H NMR (400 MHz, CDCl ₃ ) δ 7.41 (s, 1H), 6.48 (t, 1H), 4.37 (q, J=7.1 Hz, 2H), 3.75 (t, J=4.7 Hz, 4H), 3.42 (q, J=6.0 Hz, 2H), 2.54-2.43 (m, 6H), 1.83 (p, J=6.4 Hz, 2H), 1.39 (t, J=7.1 Hz, 3H).

Step B: ethyl 2-[(6-chloro-5-methylpyridazin-3-yl)[3-(morpholin-4-yl)propyl]amino]-1,3-thiazol-4carboxylate.

Sodium hydride (60% dispersion; 350 mg, 8.75 mmol, 1.31 eq.) under nitrogen was added to a solution of the product from step A (2 g, 6.68 mmol, 1 eq.) and 3,6-dichloro-4-methylpyridazine (1.25 g, 7.67 mmol, 1.15 eq.) in 1,4-dioxane (30 ml) at 0°C, and the mixture was stirred for 1 hour at 0°C and then for 3 hours at 60°C. The solution was allowed to cool to ambient temperature, then partitioned between ethyl acetate (300 ml) and saturated aqueous ammonium chloride (100 ml). The organic phase was washed successively with water (150 ml) and brine (100 ml), dried (magnesium sulfate) and concentrated in vacuo. Purification by flash column chromatography (50 g silica gel) eluting with ethyl acetate gave a brown oil. Trituration with heptane (15 ml) provided a solid, which was collected by filtration, washed with heptane (20 ml) and dried in vacuo to give the title product as a pale brown solid (1.5 g, 3.52 mmol, 53%).

LC/MS (C ₁₈ H ₂₄ ClN ₅ O ₃ S) 426 [M+H] ⁺ ; RT 0.96 (LCMS-V-BI).

1H NMR (400 MHz, CDCl ₃ ) δ 7.83 (s, 1H), 7.56 (d, J=1.1 Hz, 1H), 4.51 (t, 2H), 4.40 (q, J=7.1 Hz, 2H), 3.75 ( t, J=4.6 Hz, 4H), 2.52-2.42 (m, 9H), 2.06 (p, 2H), 1.41 (t, J=7.1 Hz, 3H).

Step C: ethyl 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}[3-(morpholin-4yl)propyl]amino)-1,3- thiazole-4-carboxylate.

The product from step B (900 mg, 2.11 mmol, 1 eq.) and 2-aminobenzothiazole (350 mg, 2.33 mmol, 1.1 eq.) were added to a solution of tris(dibenzylideneacetone)dipalladium(0) (96.8 mg, 0.11 mmol) under a nitrogen atmosphere , 0.05 eq.) and xantphos (122 mg, 0.21 mmol, 0.1 eq.) in 1,4-dioxane (20 ml), and the mixture was stirred. H^diisopropylethylamine (1.1 mL, 6.34 mmol, 3 eq.) was added and the mixture was heated in a sealed tube at 150°C for 24 hours. The solution was allowed to cool to ambient temperature and concentrated in vacuo. Purification by flash column chromatography (50 g silica gel) eluting with 10:1 dichloromethane/methanol resulted in a red/brown gum. Trituration with diethyl ether (20 ml) provided a solid which was collected by filtration, washed with diethyl ether (2 x 10 ml) and dried in vacuo to give the title product as a pale brown solid (875 mg, 1.62 mmol, 77%) .

- 120 046336

LC/MS (C ₂₅ H ₂₉ N ₇ O ₃ S ₂ ) 540 [M+H]+; RT 1.12 (LCMS-V-Bl).

¹ H NMR (400 MHz, DMSO^b) δ 11.16 (br s, 1H), 7.96 (s, 1H), 7.92 (s, 1H), 7.76 (s, 1H), 7.63-7.47 (m, 1H), 7.39 (t, 1H), 7.22 (t, J=7.5 Hz, 1H), 4.43 (t, J=7.4 Hz, 2H), 4.29 (q, J=7.1 Hz, 2H), 3.59 (t, J=5.0 Hz, 4H), 2.48 (s, 3H), 2.43-2.33 (m, 6H), 1.99-1.85 (m, 2H), 1.32 (t, J=7.1 Hz, 3H).

Step D: 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}[3-(morpholin-4yl)propyl]amino)-1,3-thiazole -4-carboxylic acid.

To a solution of the product from step C (300 mg, 0.56 mmol, 1 eq.) in a 3:1 mixture of 1,4-dioxane/water (4 ml) was added lithium hydroxide monohydrate (93.3 mg, 2.22 mmol, 4 eq.), and the mixture was stirred at ambient temperature overnight. Water (5 ml) was added and the mixture was filtered through celite and washed with water (10 ml). The filtrate was acidified with acetic acid and stirred for 15 min, then the solids were collected by filtration, washed with water (10 mL) and dried in vacuo to give the title product as a yellow solid (180 mg, 0.35 mmol, 63%).

MSHR-ERI (m/z) [MH], calculated for C ₂₃ H ₂₄ N ₇ O ₃ S ₂ : 510.1388; found: 510.1406.

Example 45: 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl} (propyl)amino)-1,3 thiazol-4-carboxylic acid

Step A: ethyl 2-(propylamino)-1,3-thiazole-4-carboxylate.

Ethyl bromopyruvate (4 ml, 28.7 mmol, 1.13 eq.) was added under nitrogen atmosphere to a solution of propylthiourea (3 g, 25.4 mmol, 1 eq.) in ethanol (50 ml) at 0°C, and the mixture was stirred for 1 h at environment. Triethylamine (7 mL, 50.8 mmol, 2 eq.) was added and the mixture was stirred for 18 hours. The reaction mixture was partitioned between ethyl acetate (300 mL) and water (150 mL) and the organic phase was washed successively with water (150 mL) and brine (100 ml), dried (magnesium sulfate) and concentrated in vacuo. Purification by flash column chromatography (50 g silica gel) eluting with dichloromethane gave the title product as a pale brown oil which solidified on standing (3.4 g, 15.9 mmol, 63%).

LC/MS (C ₉ H ₁₄ N ₂ O ₂ S) 215 [M+H]+; RT 1.04 (ZHMS-U-V1).

¹ H NMR (400 MHz, CDCl ₃ ) δ 7.42 (s, 1H), 5.63 (s, 1H), 4.37 (q, 2H), 4.40 (q, J=7.1 Hz, 2H), 3.24 (q, 2H) , 1.75-1.64 (m, 2H), 1.39 (t, 3H), 1.00 (t, J=7.1 Hz, 3H).

Step B: ethyl 2-[(6-chloro-5-methylpyridazin-3-yl)(propyl)amino]-1,3-thiazol-4-carboxylate.

Sodium hydride (60% dispersion; 500 mg, 12.5 mmol, 1.25 eq.) under nitrogen was added to a solution of the product from step A (2.15 g, 10.0 mmol, 1 eq.) and 3,6-dichloro-4-methylpyridazine (1.75 g, 10.7 mmol, 1.07 eq.) in 1,4-dioxane (30 ml) at 0°C, and the mixture was stirred for 1 hour at 0°C and then for 18 hours at ambient temperature. The reaction mixture was partitioned between ethyl acetate (300 ml) and saturated aqueous ammonium chloride (150 ml), and the organic phase was successively washed with water (2x150 ml) and brine (100 ml), dried (magnesium sulfate) and concentrated in vacuo. Purification by flash column chromatography (100 g silica gel) eluting with dichloromethane provided a solid which was triturated with heptane (30 ml), filtered, washed with heptane (15 ml) and dried in vacuo to give the title product as a white solid (1.3 g, 3.81 mmol, 38%).

LC/MS (C ₁₄ H ₁₇ ClN ₄ O ₂ S) 341 [M+H]+; RT 1.32 (LC]^^^).

¹ H NMR (400 MHz, CDCl ₃ ) δ 7.82 (s, 1H), 7.39 (d, 1H), 4.47-4.35 (m, 4H), 2.48 (d, J=1.0 Hz, 3H), 1.931.78 ( m, 2H), 1.42 (t, J=7.1 Hz, 3H), 1.05 (t, J=7.4 Hz, 3H).

Step C: ethyl 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(propyl)amino)-1,3thiazol-4-carboxylate.

The product from step B (850 mg, 2.49 mmol, 1 eq.) and 2-aminobenzothiazole (425 mg, 2.83 mmol, 1.13 eq.) were added to a solution of tris(dibenzylideneacetone)dipalladium(0) (114 mg, 0.12 mmol) under a nitrogen atmosphere , 0.05 eq.) and xantphos (144 mg, 0.25 mmol, 0.1 eq.) in 1,4-dioxane (10 ml), and the mixture was stirred. Na-diisopropylethylamine (1.3 mL, 7.48 mmol, 3 eq.) was added and the mixture was heated in a sealed tube at 150°C for 24 h. The solution was allowed to cool to ambient temperature and concentrated in vacuo. Purification by flash column chromatography (50 g silica gel) eluting with 39:1 dichloromethane/methanol gave a dark yellow solid, which was triturated with methanol (15 ml), filtered, washed with methanol (15 ml) and dried in under vacuum to obtain the desired product as a yellow solid (915 mg, 2.01 mmol, 81%).

- 121 046336

LC/MS (C21H22N6O2S2) 455 [M+H]+; RT 1.44 (LCMS-V-Bl).

1H NMR (400 MHz, DMSO-de) δ 11.12 (br s, 1H), 7.96 (s, 1H), 7.92 (br s, 1H), 7.73 (s, 1H), 7.59 (br s, 1H), 7.39 (t, J=7.5 Hz, 1H), 7.22 (t, J=7.3 Hz, 1H), 4.37 (dd, J=9.0, 6.3 Hz, 2H), 4.29 (q, J=7.1 Hz, 2H), 2.48 (s, 3h), 1.74 (q, J=7.5 Hz, 2H), 1.32 (t, J=7.1 Hz, 3H), 0.96 (t, J=7.4 Hz, 3H).

Step D: 2-({6-[( 1,3-benzothiazol-2-yl)amino]-5 -methylpyridazin-3 -yl}(propyl)amino)-1,3-thiazol4-carboxylic acid.

To a solution of the product from step C (250 mg, 0.55 mmol, 1 eq.) in a 3:1 mixture of 1,4-dioxane/water (4 ml) was added lithium hydroxide monohydrate (92.3 mg, 2.2 mmol, 4 eq.), and the mixture was stirred at ambient temperature for 18 hours. Water (5 ml) was added and the solution was filtered through celite and washed with water (10 ml). The filtrate was acidified with acetic acid to give a suspension which was collected by filtration, the crude product was washed with water (10 ml) and dried in vacuo to give the title product as a pale yellow solid (185 mg, 0.43 mmol, 79%).

MSVR-ERI (m/z) [MH] ^- , calculated for C ₁₉ H ₁₇ N ₆ O ₂ S ₂ : 425.0860; found: 425.0867.

Example 46: 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}[3-(diethylamino)propyl]amino)-1,3-thiazol-4 -carboxylic acid

Step A: ethyl 2-{[3-(diethylamino)propyl]amino}-1,3-thiazole-4-carboxylate.

Ammonia (2 M in isopropanol; 60 mL, 0.12 mol, 3.45 eq.) was added at 0°C to a solution of 3(diethylamino)propyl isothiocyanate (6 g, 34.8 mmol, 1 eq.) in isopropanol (15 mL), and the mixture stirred for 1 hour at 0°C and then for 18 hours at ambient temperature. The suspension was concentrated in vacuo, then triturated with heptane (50 ml), the solids were filtered and washed with heptane (2x25 ml). Ethanol (150 mL) and ethyl bromopyruvate (5 mL, 39.8 mmol, 1.14 eq.) were added and the mixture was stirred under nitrogen for 1 h. Triethylamine (10 mL, 69.7 mmol, 2 eq.) was added and the mixture was heated at 85 °C for 4 hours. The solution was allowed to cool to ambient temperature and concentrated in vacuo. Purification by flash column chromatography (100 g silica gel) eluting with 19:1 dichloromethane/methanol gave the title product as a dark brown gum (3.95 g, 13.8 mmol, 40%).

LC/MS (C13H23N3O2S) 286 [M+H]+; RT 0.78 (LCMS-V-BI).

1H NMR (400 MHz, CDC13) δ 7.39 (s, 1H), 7.05 (s, 1H), 4.37 (q, J=7.1 Hz, 2H), 3.42 (td, J=6.2, 3.1 Hz, 2H), 2.64 -2.51 (m, 6H), 1.84-1.76 (m, 2H), 1.39 (t, J=7.1 Hz, 3H), 1.06 (t, J=7.1 Hz, 6H).

Step B: ethyl 2-[(6-chloro-5-methylpyridazin-3-yl)[3-(diethylamino)propyl]amino]-1,3-thiazol-4carboxylate.

Sodium hydride (60% dispersion; 700 mg, 17.5 mmol, 1.26 eq.) under nitrogen was added to a solution of the product from step A (3.95 g, 13.8 mmol, 1 eq.) and 3,6-dichloro-4-methylpyridazine (2.5 g, 15.3 mmol, 1.11 eq.) in 1,4-dioxane (50 ml) at 0°C, and the mixture was stirred for 1 hour at 0°C and then for 18 hours at ambient temperature. The reaction mixture was partitioned between ethyl acetate (300 ml) and water (150 ml), and the organic phase was washed successively with water (150 ml) and brine (100 ml), dried (magnesium sulfate) and concentrated in vacuo. Purification by flash column chromatography (100 g silica gel) eluting with 19:1 dichloromethane/methanol gave the title product as a brown gum (2.3 g, 5.58 mmol, 40%).

LC/MS (C ₁₈ H ₂₆ ClN ₅ O ₂ S) 412 [M+H]+; RT 1.00 (LCMS-V-BI).

1H NMR (400 MHz, CDCl ₃ ) δ 7.80 (s, 1H), 7.63 (d, J=1.1 Hz, 1H), 4.47 (q, 2H), 4.37 (q, J=7.1 Hz, 2H), 2.64- 2.54 (m, 6H), 2.45 (d, J=0.9 Hz, 3H), 1.98 (q, 2H), 1.39 (t, J=7.1 Hz, 3H), 1.06 (t, J=7.1 Hz, 6H).

Step C: ethyl 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}[3(diethylamino)propyl]amino)-1,3-thiazol-4 -carboxylate.

The product from step B (2.1 g, 5.1 mmol, 1 eq.) and 2-aminobenzothiazole (850 mg, 5.66 mmol, 1.11 eq.) were added to a solution of tris(dibenzylideneacetone)dipalladium(0) (233 mg, 0.25 mmol) under a nitrogen atmosphere , 0.05 eq.) and xantphos (295 mg, 0.51 mmol, 0.1 eq.) in 1,4-dioxane (20 ml), and the mixture was stirred. N,N-diisopropylethylamine (2.7 mL, 15.3 mmol, 3 eq.) was added and the mixture was heated in a sealed tube at 150°C for 24 hours. The solution was allowed to cool to ambient temperature and concentrated in vacuo. Purification by flash column chromatography (100 g silica gel) eluting with

- 122 046336 a gradient of 5-10% methanol in dichloromethane gave a dark yellow gum, which was triturated with diethyl ether (10 ml), filtered, washed with diethyl ether (10 ml) and dried in vacuum to obtain the desired product as a yellow solid substances (240 mg, 0.46 mmol, 9%).

LC/MS (C ₂₅ H ₃₁ N ₇ O ₂ S ₂ ) 526 [M+H]+; RT 1.18 (LCMS-V-BI).

¹ H NMR (400 MHz, DMSO-dg) δ 11.78 (br s, 1H), 7.88-7.67 (m, 4H), 7.44-7.34 (m, 1H), 7.26-7.18 (m, 1H), 4.49 (s , 2H), 4.36 (q, J=7.1 Hz, 2H), 3.31-3.22 (m, 2H), 3.22-3.12 (m, 4H), 2.52-2.39 (m, 5H), 1.39 (t, J=7.1 Hz, 9H).

Step D: 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}[3-(diethylamino)propyl]amino)-1,3-thiazol-4 -carboxylic acid.

To a solution of the product from step C (200 mg, 0.38 mmol, 1 eq.) in a 3:1 mixture of 1,4-dioxane/water (4 ml) was added lithium hydroxide monohydrate (63.9 mg, 1.52 mmol, 4 eq.), and the mixture was stirred at ambient temperature for 18 hours. Water (5 ml) was added and the mixture was filtered through celite, washing with water (10 ml). The filtrate was acidified with acetic acid to give a solid, which was collected by filtration, washed with water (10 mL) and dried in vacuo to give the title product as a pale yellow solid (105 mg, 0.21 mmol, 55%).

MSVR-ERI (m/z) [MH] ^- , calculated for C ₂₃ H ₂₆ N ₇ O ₂ S ₂ : 496.1595; found: 496.1614.

Example 47: 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(3-methoxypropyl)amino)1,3-thiazol-4-carboxylic acid

Lithium hydroxide monohydrate (104 mg, 2.48 mmol, 4 equiv.) was added to a solution of synthesis product 11c, step C (300 mg, 0.62 mmol, 1 eq.) in a 3:1 mixture of 1,4-dioxane/water (4 ml). ), and the mixture was stirred at ambient temperature overnight. Water (15 mL) was added and the mixture was acidified with acetic acid to give a solid, which was collected by filtration, washed with water (25 mL) and dried in vacuo to give the title product as an orange solid (245 mg, 0.54 mmol, 87% ).

MSHR-ERI (m/z) [M+H]+, calculated for C20H21N6O3S2: 457.1111; found: 457.1119.

Example 48: 2-[[4,5-Dimethyl-6-[(4-methyl-1,3-benzothiazol-2-yl)amino]pyridazin-3-yl]amino]thiazole4-carboxylic acid

Step A: N(6-chloro-4,5-dimethylpyridazin-3-yl)-4-methyl-1,3-benzothiazol-2-amine.

To 611 mg of 6-chloro-4,5-dimethylpyridazin-3-amine (3.9 mmol, 1 eq.) and 884 mg of 2-bromo-4-methyl-1,3benzothiazole (3.9 mmol, 1 eq.) in 13 ml 1 ,4-dioxane was added 1.5 g of sodium hydride (60 wt.%, 37.5 mmol, 9.6 eq.), and the mixture was stirred at reflux for 1 hour. After cooling, the reaction mixture was poured into ice water and acidified with 2N. HCl solution. The precipitate was filtered off, washed with NaHCO ₃ solution, and dried to obtain 810 mg (67%) of the target product.

LC-MS-ESI (m/z): [M+H]+, calculated for C ₁₄ H ₁₄ ClN ₄ S: 305; found: 305.

Step B: ethyl 2-[[4,5-dimethyl-6-[(4-methyl-1,3-benzothiazol-2-yl)amino]pyridazin-3yl]amino]thiazol-4-carboxylate.

A mixture of 600 mg of the product from step A (1.96 mmol, 1 eq.), 730 mg of ethyl 2-aminothiazole-4-carboxylate (4.24 mmol, 2.1 eq.), 78 mg of Pd ₂ (dba) ₃ (0.085 mmol, 0.04 eq. ), 124 mg of 4,5-bis(diphenylphosphino)-9,9dimethylxanthene (0.21 mmol, 0.1 equiv.) and 2.8 ml of DIPEA (8 equiv.) in 16 ml of 1,4-dioxane were stirred at 185°C in a microwave reactor in for 1.5 h. After cooling, volatiles were removed and the crude intermediate was purified by flash column chromatography using heptane and

- 123 046336

EtOAc as eluents to obtain 328 mg (38%) of the desired product.

Ή NMR (500 MHz, DMSO-de) δ ppm 11.00/10.81 (s+brs, 2H), 7.94 (s, 1H), 7.72 (brs, 1H), 7.19 (d, 1H), 7.10 (t, 1H), 4.29 (q, 2H), 2.57 (brs, 3H), 2.39/2.36 (s+s, 6H), 1.31 (t, 3H); ¹³ C NMR (125 MHz, DMSO^) δ ppm 126.9, 122.4, 121.7, 119.4, 60.8, 18.4, 14.7; MSHR-ERI (m/z): [M+H] ⁺ , calculated for C ₂₀ H ₂₀ N ₆ O ₂ S ₂ : 441.1167; found: 441.1165.

Step C: 2-[[4,5-dimethyl-6-[(4-methyl-1,3-benzothiazol-2-yl)amino]pyridazin-3-yl]amino]thiazol-4carboxylic acid.

A mixture of the product from stage B, LiOHxH ₂ O, water and 1,4-dioxane was stirred at reflux for 1 hour. After cooling and concentration, 1N was added. HCl, the precipitate was filtered off and further purified using reverse phase preparative chromatography to obtain the desired product.

MSHR-ERI (m/z): [M+H] ⁺ , calculated for C ₁₈ H ₁₇ N ₆ O ₂ S ₂ : 413.0854; found: 413.0855.

Example 49: 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(3-hydroxypropyl)amino)-1,3-thiazol-4-carboxylic acid

Step A: ethyl 2-[(6-chloro-5-methylpyridazin-3-yl)(3-hydroxypropyl)amino]-1,3-thiazol-4carboxylate.

Boron tribromide (1 M in dichloromethane; 16 ml, 16.2 mmol, 2 eq.) under nitrogen atmosphere was added to a solution of the product of synthesis 11c, step B (3 g, 8.09 mmol, 1 eq.), in dichloromethane (50 ml) at - 78°C. The mixture was stirred for 1 hour at -78°C, then heated at 50°C for 1 hour. The suspension was allowed to cool to ambient temperature, then partitioned between dichloromethane (250 ml) and saturated aqueous sodium hydrogen carbonate (150 ml). The organic phase was washed successively with water (2x150 ml) and brine (100 ml), dried (magnesium sulfate) and concentrated in vacuo. Purification by flash column chromatography (50 g silica gel) eluting with 39:1 dichloromethane/methanol gave a solid which was triturated with diethyl ether (15 ml), filtered, washed with diethyl ether (20 ml) and dried in vacuo with yielding the desired product as an orange solid (900 mg, 2.52 mmol, 31%).

LC/MS (C ₁₄ H ₁₇ ClN ₄ O ₃ S) 357 [M+H] ⁺ ; RT 1.15 (LCMS-V-BI).

1H NMR (400 MHz, DMSOD6) δ 8.04 (s, 1H), 7.87 (dd, J=18.1, 1.1 Hz, 1H), 4.71 (td, J=5.4, 1.1 Hz, 1H), 4.46 (t, 2H) , 4.29 (q, J=7.1 Hz, 2H), 3.50 (q, J=5.9 Hz, 2H), 2.43 (dd, J=2.6, 0.9 Hz, 3H), 1.89 (p, J=6.5 Hz, 2H) , 1.31 (t, J=7.1 Hz, 3H).

Step B: ethyl 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(3hydroxypropyl)amino)-1,3-thiazol-4-carboxylate.

The product from step A (500 mg, 1.4 mmol, 1 eq.) and 2-aminobenzothiazole (240 mg, 1.6 mmol, 1.14 eq.) were added to a solution of tris(dibenzylideneacetone)dipalladium(0) (64.2 mg, 0.07 mmol) under a nitrogen atmosphere , 0.05 eq.) and xantphos (81.1 mg, 0.14 mmol, 0.1 eq.) in 1,4-dioxane (10 ml), and the mixture was stirred. HX-diisopropylethylamine (0.75 ml, 4.2 mmol, 3 eq.) was added and the mixture was heated in a sealed tube at 150°C for 24 hours. The solution was allowed to cool to ambient temperature and concentrated in vacuo. Purification by flash column chromatography (50 g silica gel) eluting with 19:1 dichloromethane/methanol gave a red/orange gum which was triturated with diethyl ether (15 ml), filtered, washed with diethyl ether (2x10 ml) and dried in vacuo to give the title product as an orange solid (490 mg, 0.94 mmol, 67%).

LC/MS (C ₂₁ H ₂₂ N ₆ O ₃ S ₂ ) 471 [M+H] ⁺ ; RT 1.31 (LC\1С-\'-В1).

1H NMR (400 MHz, DMSOD6) δ 11.12 (br s, 1H), 7.98 (s, 1H), 7.92 (br s, 1H), 7.72 (s, 1H), 7.63 (br s, 1H), 7.40 (t , J=7.6 Hz, 1H), 7.23 (t, 1H), 4.74 (t, J=5.4 Hz, 1H), 4.46 (t, J=7.1 Hz, 2H), 4.30 (q, J=7.1 Hz, 2H ), 3.54-3.46 (m, 2H), 2.48 (s, 3H), 1.90 (p, J=6.5 Hz, 2H), 1.32 (t, J=7.1 Hz, 3H).

Step C: 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(3-hydroxypropyl)amino)1,3-thiazol-4-carboxylic acid.

To a solution of the product from step B (275 mg, 0.53 mmol, 1 eq.) in a 3:1 mixture of 1,4-dioxane/water (4 ml) was added lithium hydroxide monohydrate (88.3 mg, 2.1 mmol, 4 eq.), and the mixture was stirred at ambient temperature for 18 hours. Water (15 ml) was added and the mixture was filtered through celite. Filt

- 124 046336 the rat was acidified with acetic acid to give a solid, which was collected by filtration, washed with water (30 ml) and dried in vacuo to give the title product as a yellow solid (190 mg, 0.43 mmol, 82%).

HRMS-ERI (m/z) [M+H]+, calculated for C ₁₉ H ₁₉ N ₆ O ₃ S ₂ : 443.0955; found: 443.0996.

Example 50: 2-[[6-(1,3-benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]methylamino]-5-[3-[4-[3(dimethylamino)prop-1-ynyl ]-2-fluorophenoxy]propyl]thiazole-4-carboxylic acid

Step A: ethyl 5-[3-[4-[3-(dimethylamino)prop-1-ynyl]-2-fluorophenoxy]propyl]-2-[methyl-[5-methyl6-[^)-[3-( 2-trimethylsilylethoxymethyl)-1,3-benzothiazol-2-ylidene]amino]pyridazin-3-yl]amino]thiazol4-carboxylate.

Using a general alkylation procedure, starting with synthesis 5g and synthesis 6b as a suitable phenol, the desired product was obtained.

1H NMR (500 MHz, DMSO-dg) δ ppm 7.84 (d, 1H), 7.67 (s, 1H), 7.47 (d, 1H), 7.44 (t, 1H), 7.33 (dd, 1H), 7.25 (t, 1H), 7.22 (dd, 1H), 7.16 (t, 1H), 5.86 (s, 2H), 4.26 (q, 2H), 4.15 (t, 2H), 3.77 (s, 3H), 3.72 (t, 2H), 3.49 (brs, 2H), 3.27 ( t, 2H), 2.46 (s, 3H), 2.27 (s, 6H), 2.13 (qn, 2H), 1.29 (t, 3H), 0.92 (t, 2H), -0.11 (s, 9H); ¹³ C NMR (125 MHz, DMSO-d^ δ ppm 129.0, 127.2, 123.5, 123.2, 119.2, 117.7, 115.5, 111.9, 72.8, 68.5, 66.7, 60.7, 48.2, 44.0, 35 .3, 31.1, 23.2, 17.9, 17.8, 14.6, -0.9; MSHR-ERI (m/z): [M+H] ⁺ , calculated for C ₃₉ H ₄₉ FN ₇ O ₄ S ₂ Si: 790.3035; found: 790.3023.

Step B: 2-[[6-(1,3-benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]methylamino]-5-[3-[4-[3(dimethylamino)prop-1-ynyl ]-2-fluorophenoxy]propyl]thiazole-4-carboxylic acid.

Using a general deprotection and hydrolysis procedure, starting with the product from Step A as a suitable ethyl ester, the desired product was obtained.

MSVR-ERI (m/z): [M+H] ⁺ , calculated for C ₃₁ H ₃₁ FN ₇ O ₃ S ₂ : 632.1908; found: 632.1913.

Example 51: 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methyl-4-[3-(4-methylpiperazin-1yl)propyl]pyridazin-3-yl} amino) -1,3-thiazole-4-carboxylic acid] Η _Λ \>Λζ ^Ν γ ^Ν ._I xC. _Χ Ν OH

Step A: ethyl 2-({6-[(1,3-benzothiazol-2-yl)amino]-4-[3-(benzyloxy)propyl]-5-methylpyridazin-3yl}amino)-1,3-thiazole -4-carboxylate.

Synthesis product 9ma (500 mg, 1.18 mmol, 1 eq.), ethyl 2-amino-1,3-thiazol-4-carboxylate (284 mg, 1.65 mmol, 1.4 eq.) was loaded into a microwave-dried vessel for microwave treatment. , cesium carbonate (575 mg, 1.76 mmol, 1.5 eq.), xanthos (68.1 mg, 0.12 mmol, 0.1 eq.) and tris(dibenzylideneacetone) diialladium(0) (53.9 mg, 0.06 mmol, 0.05 eq.). The vessel was evacuated and purged with nitrogen (x3), and then alpha,alpha,alpha-trifluorotoluene (20 ml) was added. The mixture was bubbled with nitrogen (10 min) and then heated at 180°C for 1 h under microwave irradiation. The reaction mixture was concentrated in vacuo and purified by automated flash column chromatography (CombiFlash Rf, 40 g RediSep™ silica gel cartridge) eluting with a gradient of 0-70% ethyl acetate in isoheptane to give the title product as a yellow solid (336 mg, 0.6 mmol, 51%).

LC/MS (C ₂₈ H ₂₈ N ₆ O ₃ S ₂ ) 561 [M+H] ⁺ ; RT 1.53 (LCMS-V-BI).

1H NMR (400 MHz, DMSO-dg) δ 11.08 (br s, 1H), 7.95 (d, 1H), 7.88 (br s, 1H), 7.40-7.33 (m, 6H), 7.327.26 (m, 1H ), 7.20 (t, 1H), 4.51 (s, 2H), 4.30 (q, J=7.1 Hz, 2H), 3.54 (t, J=6.2 Hz, 2H), 2.96 (t, J=7.8 Hz, 2H ), 2.39 (s, 3H), 1.76 (p, 2H), 1.32 (t, J=7.1 Hz, 3H).

- 125 046336

Step B: ethyl 2-({6-[(1,3-benzothiazol-2-yl)amino]-4-(3-hydroxypropyl)-5-methylpyridazin-3yl}amino)-1,3-thiazol-4- carboxylate.

To a cooled solution of the product from step A (330 mg, 0.59 mmol, 1 eq.) in anhydrous dichloromethane (15 ml), boron trichloride (1 M in dichloromethane; 1.18 ml, 1.18 mmol, 2 eq.) was added dropwise and the mixture was stirred for 3 hours. The reaction mixture was cooled to 0°C and quenched by adding saturated aqueous sodium bicarbonate. The mixture was diluted with dichloromethane and the layers were separated. The organic phase was washed successively with water and brine, dried (magnesium sulfate) and concentrated in vacuo to give the title product (301 mg, 0.64 mmol, >100%) as a mixture with the appropriate methyl ester, which was directly used in the next step without further cleaning.

LC/MS (C ₂₁ H ₂₂ N ₆ O ₃ S ₂ ) 471 [M+H]+; RT 1.295 (ZHMS-U-B1).

Step C: ethyl 2-{[4-(3-hydroxypropyl)-5-methyl-6-{[(22)-3-{[2-(trimethylsilyl)ethoxy]methyl}2,3-dihydro-1,3 -benzothiazol-2-ylidene]amino}pyridazin-3-yl]amino}-1,3-thiazol-4-carboxylate.

To a cooled solution of the product from step B (300 mg, 0.64 mmol, 1 eq.) in dichloromethane (15 ml) was added 2-(trimethylsilyl)ethoxymethyl chloride (0.12 ml, 0.7 mmol, 1.1 eq.), N,Ndiisopropylethylamine (0.22 ml , 1.28 mmol, 2 equiv.) and DMAP (3.89 mg, 0.03 mmol, 0.05 equiv.). The mixture was stirred for 2 hours at ambient temperature, then cooled to 0°C and quenched by adding aqueous sodium bicarbonate. The layers were separated and the organic phase was washed with water followed by brine, dried (magnesium sulfate) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 24 g RediSep™ silica gel cartridge) eluting with a gradient of 0-70% ethyl acetate in isoheptane gave the title product as an orange solid (105 mg, 0.17 mmol, 27%) , which was found to be a mixture with the corresponding methyl ester. The mixture was directly used in the next step without further purification.

LC/MS (C ₂₇ H ₃₆ N ₆ O ₄ SiS ₂ ) 601 [M+H]+; RT 1.59 (ZHMS-U-B1).

Step D: ethyl 2-{[5-methyl-4-(3-oxopropyl)-6-{[(22)-3-{[2-(trimethylsilyl)ethoxy]methyl}-2,3dihydro-1,3- benzothiazol-2-ylidene]amino}pyridazin-3-yl]amino}-1,3-thiazol-4-carboxylate.

The oven-dried flask was charged with dimethyl sulfoxide (29.6 µl, 0.42 mmol, 2.5 eq) in dichloromethane (5 ml), and the stirred solution was cooled to -78°C. A solution of oxalyl chloride (2 M; 91.5 μl, 0.18 mmol, 1.1 eq.) was added dropwise and the mixture was stirred for 1 h. A solution of the product from step C (100 mg, 0.17 mmol, 1 eq.) in dichloromethane was added dropwise ( 5 ml), and the resulting mixture was stirred at -78°C for 1 hour. Triethylamine (0.14 ml, 1 mmol, 6 eq.) was added and the mixture was allowed to warm to 0°C for 1 hour. The reaction mixture was quenched water (10 ml), diluted with sodium bicarbonate (10 ml) and extracted with dichloromethane (2x 50 ml). The combined organic extracts were washed with brine (50 ml), dried (magnesium sulfate) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 12 g RediSep™ silica gel cartridge) eluting with a gradient of 0-50% ethyl acetate in isoheptane gave the title product as an orange glass (68 mg, 0.11 mmol, 68%) , which was found to be a mixture with the corresponding methyl ester, which was directly used in the next step without further purification.

LC/MS (C ₂₇ H ₃₄ N ₆ O ₄ SiS ₂ ) 599 [M+H]+; RT 1.62 (ZHMS-U-B1).

Step E: ethyl 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methyl-4-[3-(4-methylpiperazin-1yl)propyl]pyridazin-3-yl} amino )-1,3-thiazole-4-carboxylate.

To a solution of the product from step D (68 mg, 0.11 mmol, 1 eq.) in 3:1 methanol/acetic acid (8 ml) was added 1-methylpiperazine (25.2 μl, 0.23 mmol, 2 eq.) followed by the addition of sodium cyanoborohydride (14.3 mg, 0.23 mmol, 2 eq.), and the mixture was stirred at ambient temperature for 40 minutes. The reaction mixture was quenched by adding 1 M aqueous sodium hydroxide solution and the mixture was extracted with dichloromethane (3x50 ml). The combined organic extracts were washed with water followed by brine (50 ml), dried (magnesium sulfate) and concentrated in vacuo. The residue was dissolved in dichloromethane (5 ml), trifluoroacetic acid (1 ml) was added and the mixture was stirred for 1 hour. The mixture was diluted with dichloromethane (20 ml) and washed with 1 M aqueous sodium hydroxide solution (20 ml). The organic phase was washed with brine, dried (magnesium sulfate) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 12 g RediSep™ silica gel cartridge) eluting with a 0-20% 7N gradient. A methanol solution of ammonia in dichloromethane gave the desired product as an orange glassy substance (40 mg, 0.07 mmol, 64%) which was found to be a mixture with the corresponding methyl ester, which was directly used in the next step without further purification.

LC/MS (C ₂₆ H ₃₂ N ₈ O ₂ S ₂ ) 553 [M+H]+; RT 1.17 (ZHMS-U-B1).

Step F: 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methyl-4-[3-(4-methylpiperazine-1

- 126 046336 yl)propyl]pyridazin-3 -yl} amino)-1,3-thiazol-4-carboxylic acid.

To a solution of the product from step E (40 mg, 0.07 mmol, 1 eq.) in 1,4-dioxane (5 ml) was added a 2 M aqueous solution of lithium hydroxide (0.14 ml, 0.14 mmol, 2 eq.), and the mixture was heated with reflux for 1 hour. The reaction mixture was concentrated in vacuo, then hydrochloric acid (4 M in 1,4-dioxane) was added and the mixture was stirred for 1 hour. The mixture was concentrated in vacuo and triturated with diethyl ether, filtered, washed with diethyl ether and dried in vacuo to give the title product as a dark yellow solid (14.4 mg, 0.03 mmol, 38%) [as hydrochloric acid bisalt].

HRMS-ERI (m/z) [M+H]+, calculated for C ₂₄ H ₂₉ N ₈ O ₂ S ₂ : 525.1849; found: 525.1888.

Example 52: 5-{1-[(adamantan-1-yl)methyl]-5-methyl-1H-pyrazol-4-yl}-2-({6-[(1,3-benzothiazol-2yl)amino] -5-methylpyridazin-3-yl}(3-methoxypropyl)amino)-1,3-thiazol-4-carboxylic acid

Step A: ethyl 5-{1-[(adamantan-1-yl)methyl]-5-methyl-1H-pyrazol-4-yl}-2-[(3-methoxypropyl)(5methyl-6-{[(2Z )-3-{[2-(trimethylsilyl)ethoxy]methyl}-2,3-dihydro-1,3-benzothiazol-2ylidene]amino}pyridazin-3-yl)amino]-1,3-thiazol-4-carboxylate .

To a mixture of synthesis product 11c (100 mg, 0.14 mmol, 1 eq.), synthesis product 10a (61.6 mg, 0.17 mmol, 1.2 eq.) and potassium carbonate (59.8 mg, 0.43 mmol, 3 eq.) in a 4:1 mixture tetrahydrofuran/water (5 ml), [1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium(II) (10.6 mg, 0.01 mmol, 0.1 eq.) was added. The mixture was bubbled with nitrogen (10 min) and then heated at 120°C for 1 h under microwave irradiation. The reaction mixture was diluted with ethyl acetate, washed with water followed by brine, dried (magnesium sulfate) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 12 g RediSep™ silica gel cartridge) eluting with a gradient of 0-50% ethyl acetate in isoheptane gave the title product as a creamy foam (89.2 mg, 0.11 mmol, 73%).

LC/MS (C ₄₃ H ₅₈ N ₈ O ₄ SiS ₂ ) 843 [M+H] ⁺ ; RT 1.88 (ZHMS-U-V2).

¹ H NMR (400 MHz, DMSO^) δ 7.82 (dd, J=7.7, 1.0 Hz, 1H), 7.70 (d, J=1.1 Hz, 1H), 7.52 (s, 1H), 7.487.40 (m, 2H), 7.23 (ddd, J=8.3, 7.0, 1.5 Hz, 1H), 5.87 (s, 2H), 4.44 (t, J=7.2 Hz, 2H), 4.14 (q, J=7.1 Hz, 2H), 3.78 (s, 2H), 3.72 (t, 2H), 3.44 (t, J=6.0 Hz, 2H), 3.25 (s, 3H), 2.45 (s, 3H), 2.19 (s, 3H), 2.06-1.91 (m, 5H), 1.72-1.54 (m, 12H), 1.14 (t, J=7.1 Hz, 3H), 0.91 (t, 2H), -0.13 (s, 9H).

Step B: ethyl 5-{1-[(adamantan-1-yl)methyl]-5-methyl-1H-pyrazol-4-yl}-2-({6-[(1,3-benzothiazol-2yl)amino ]-5-methylpyridazin-3-yl} (3-methoxypropyl)amino)-1,3-thiazol-4-carboxylate.

A solution of the product from Step A (89 mg, 0.11 mmol, 1 eq.) in dichloromethane (5 mL) was cooled in an ice bath and trifluoroacetic acid (0.5 mL, 6.71 mmol, 64 eq.) was added. The mixture was allowed to warm to ambient temperature and stirred overnight. The reaction mixture was diluted with dichloromethane, washed with saturated aqueous sodium bicarbonate, dried (magnesium sulfate) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 12 g RediSep™ silica gel cartridge) eluting with a gradient of 0-5% methanol in dichloromethane gave the title product as a yellow glass (63.2 mg, 0.09 mmol, 84%) .

LC/MS (C ₃₇ H ₄₄ N ₈ O ₃ S ₂ ) 713 [M+H] ⁺ ; RT 1.48 (LCMS-V-BI).

1H NMR (400 MHz, DMSO^) δ 11.10 (br s, 1H), 7.93 (br s, 1H), 7.71 (br s + s, 2H), 7.54 (s, 1H), 7.38 (t, 1H), 7.25-7.16 (m, 1H), 4.44 (t, J=7.2 Hz, 2H), 4.16 (q, J=7.1 Hz, 2H), 3.79 (s, 2H), 3.46 (t, J=6.0 Hz, 2H ), 3.26 (s, 3H), 2.48 (s, 3H), 2.20 (s, 3H), 2.07-1.97 (m, 5H), 1.72-1.56 (m, 12H), 1.16 (t, J=7.1 Hz, 3H).

Step C: 5-{1-[(adamantan-1-yl)methyl]-5-methyl-1H-pyrazol-4-yl}-2-({6-[(1,3-benzothiazol-2yl)amino] -5-methylpyridazin-3-yl}(3-methoxypropyl)amino)-1,3-thiazol-4-carboxylic acid.

To a solution of the product from step B (63 mg, 0.09 mmol, 1 eq.) in 1,4-dioxane (5 ml) was added lithium hydroxide monohydrate (18.5 mg, 0.44 mmol, 5 eq.), and the mixture was refluxed at during the night. The reaction mixture was allowed to cool to ambient temperature and then concentrated in vacuo. The residue was triturated with water and the solids were collected by filtration, washed with water and dried in vacuo to give the title product as a yellow solid (11.5 mg, 0.02 mmol, 19%) [as lithium salt].

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MSHR-ERI (m/z) [M+H]+, calculated for C35H41N8O3S2: 685.2738; found: 685.2753.

Example 53: 5-{1-[(adamantan-1-yl)methyl]-5-methyl-1H-pyrazol-4-yl}-2-({6-[(1,3-benzothiazol-2yl)amino] -5-cyclopropylpyridazin-3-yl} (methyl)amino)-1,3-thiazol-4-carboxylic acid

Step A: ethyl 5-{1-[(adamantan-1-yl)methyl]-5-methyl-1H-pyrazol-4-yl}-2-[(5-cyclopropyl-6{[(2Z)-3- {[2-(trimethylsilyl)ethoxy]methyl}-2,3-dihydro-1,3-benzothiazol-2-ylidene]amino}niridazin-3yl)(methyl)amino] - 1,3-thiazol-4-carboxylate.

To a mixture of synthesis product 11d (149 mg, 0.23 mmol, 1 eq.), synthesis product 10a (96.3 mg, 0.27 mmol, 1.2 eq.) and potassium carbonate (93.4 mg, 0.68 mmol, 3 eq.) in a 5:2 mixture tetrahydrofuran/water (7 ml), [1,G-bis(diphenylphosphino)ferrocene]dichloropalladium(P) (16.5 mg, 0.02 mmol, 0.1 eq.) was added. The mixture was bubbled with nitrogen (10 min) and then heated at 120°C for 1 h under microwave irradiation. The reaction mixture was partitioned between ethyl acetate and water, and the organic phase was washed with brine, dried (magnesium sulfate) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 12 g RediSep™ silica gel cartridge) eluting with a gradient of 0-50% ethyl acetate in isoheptane gave the title product as a white foam (130 mg, 0.16 mmol, 71%).

LC/MS (C ₄₂ H ₅₄ N ₈ O ₃ SiS ₂ ) 811 [M+H]+; RT 1.64 (LCMS-V-BI).

¹ H NMR (400 MHz, DMSO-06) δ 7.81 (dd, J=7.6, 1.0 Hz, 1H), 7.52 (s, 1H), 7.49-7.40 (m, 2H), 7.23 (ddd, J=8.3, 7.0, 1.4 Hz, 1H), 7.10 (s, 1H), 5.88 (s, 2H), 4.15 (q, J=7.1 Hz, 2H), 3.80 (s, 3H), 3.79 (s, 2H), 3.70 ( t, 2H), 2.72-2.62 (m, 1H), 2.18 (s, 3H), 2.01-1.92 (m, 3H), 1.71-1.54 (m, 12H), 1.21-1.13 (m, 5H), 1.12- 1.07 (m, 2H), 0.90 (t, 3H), -0.13 (s, 9H).

Step B: ethyl 5-{1-[(adamantan-1-yl)methyl]-5-methyl-1H-pyrazol-4-yl}-2-({6-[(1,3-benzothiazol-2yl)amino ]-5-cyclopropylpyridazin-3-yl} (methyl)amino)-1,3-thiazol-4-carboxylate.

Trifluoroacetic acid (1 ml, 13.4 mmol, 78.9 eq.) was added to a stirred solution of the product from step A (138 mg, 0.17 mmol, 1 eq.) in dichloromethane (3 ml) and the mixture was stirred for 18 hours. The reaction mixture was partitioned between dichloromethane and saturated aqueous sodium bicarbonate, washed successively with water and brine, dried (magnesium sulfate) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 12 g RediSep™ silica gel cartridge) eluting with a gradient of 0-5% methanol in dichloromethane gave the title product as a yellow glass (104 mg, 0.15 mmol, 90%) .

LC/MS (C ₃₆ H ₄₀ N ₈ O ₂ S ₂ ) 681 [M+H]+; RT 1.45 (LCMS-V-BI).

1H NMR (400 MHz, DMSO^) δ 11.16 (br s, 1H), 7.76 (br s, 2H), 7.51 (s, 1H), 7.41-7.33 (m, 1H), 7.247.14 (m, 2H) , 4.16 (q, J=7.1 Hz, 2H), 3.81 (s, 3H), 3.78 (s, 2H), 2.18 (s, 3H), 2.00-1.92 (m, 3H), 1.72-1.53 (m, 12H ), 1.20-1.12 (m, 5H), 1.07-1.00 (m, 2H).

Step C: 5-{1-[(adamantan-1-yl)methyl]-5-methyl-1H-pyrazol-4-yl}-2-({6-[(1,3-benzothiazol-2yl)amino] -5-cyclopropylpyridazin-3-yl} (methyl)amino)-1,3-thiazol-4-carboxylic acid.

To a solution of the product from step B (104 mg, 0.15 mmol, 1 eq.) in 1,4-dioxane (5 ml) was added lithium hydroxide monohydrate (32 mg, 0.76 mmol, 5 eq.) and the mixture was refluxed at during the night. The reaction mixture was allowed to cool to ambient temperature and concentrated in vacuo. The resulting solid was triturated with water, filtered and dried in vacuo to give the title product as a yellow solid (83.5 mg, 0.13 mmol, 84%) [as lithium salt].

HRMS-ERI (m/z) [M+H]+, calculated for C ₃₄ H ₃₇ N ₈ O ₂ S ₂ : 653.2475; found: 653.2475.

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Example 54: 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl} (3-methoxypropyl)amino)5-(3-{4-[3- (dimethylamino)prop-1-yn-1-yl]-2-fluorophenoxy}propyl)-1,3-thiazol-4-carboxylic acid

Stage A: ethyl 5-('3-rngpoKCnnpon-1-nH-1-nj)-2-[('3-MeTOKCnnponnj)('5-MeTnj-6-{[('2Z)-3-{[2 (trimethylsilyl)ethoxy]methyl}-2,3-dihydro-1,3-benzothiazol-2-ylidene]amino}pyridazin-3-yl)amino]1,3-thiazol-4-carboxylate.

Tetrakis(triphenylphosphine)palladium(0) (140.6 mg, 0.12 mmol, 0.1 equiv.) was added under a nitrogen atmosphere to a stirred solution of synthesis product 11c (844 mg, 1.22 mmol, 1 equiv.), propargyl alcohol (0.28 ml, 4.86 mmol, 4 equiv.), triethylamine (1.02 ml, 7.3 mmol, 6 equiv.) and copper(G) iodide (46.4 mg, 0.24 mmol, 0.2 equiv.) in dimethylformamide (20 ml), and the mixture was heated at 100°C for nights. The reaction mixture was allowed to cool to ambient temperature and then concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 40 g RediSep™ silica gel cartridge) eluting with a gradient of 0-100% ethyl acetate in isoheptane gave the title product as a brown gum (671 mg, 1 mmol, 83%).

LC/MS (C31H40N6O5SiS ₂ ) 669 [M+H]+; RT 1.43 (ZHHMSA-Sh).

¹ H NMR (400 MHz, DMSO) δ 7.88 (d, 1H), 7.73 (s, 1H), 7.50-7.41 (m, 2H), 7.26 (ddd, J=8.3, 7.0, 1.5 Hz, 1H), 5.88 (s, 2H), 4.45 (t, 2H), 4.29 (q, J=7.1 Hz, 2H), 4.17 (d, J=6.1 Hz, 2H), 3.72 (t, 2H), 3.42 (t, J= 5.9 Hz, 2H), 3.22 (s, 3H), 2.47 (s, 3H), 1.98 (p, J=6.7 Hz, 2H), 1.33 (t, J=7.1 Hz, 3H), 0.92 (t, 2H) , -0.11 (s, 9Н).

Step B: ethyl 5-(3-hydroxypropyl)-2-[(3-methoxypropyl)(5-methyl-6-{[(2Z)-3-{[2(trimethylsilyl)ethoxy]methyl} -2,3- dihydro-1,3-benzothiazol-2-ylidene]amino}pyridazin-3-yl)amino]1,3-thiazol-4-carboxylate.

A suspension of the product from step A (671 mg, 1 mmol, 1 eq.) in a 1:1 mixture of ethyl acetate/methanol (30 ml) was loaded into a flask containing platinum oxide (PO) (0.02 g, 0.1 mmol, 0.1 eq.) in nitrogen atmosphere. The mixture was evacuated and filled with nitrogen again (x3), then evacuated and placed under a hydrogen atmosphere. The mixture was shaken for 3.5 hours, then filtered through celite (10 g) and washed with methanol. The solvent was removed in vacuo and the crude product was purified by automated flash column chromatography (CombiFlash Rf, 12 g RediSep™ silica gel cartridge) eluting with a gradient of 0-60% ethyl acetate in isoheptane to give the title product as a yellow foam (431 mg, 0.64 mmol, 64%).

LC/MS (C31H44N6O5SiS2) 673 [M+H]+; RT 1.43 (ZHMS-U-V1).

^1H NMR (400 MHz, DMSO^ ₆ ) δ 7.86 (d, 1H), 7.66 (s, 1H), 7.49-7.40 (m, 2H), 7.28-7.22 (m, 1H), 5.87 (s, 2H) , 4.39 (t, J=7.2 Hz, 2H), 4.27 (q, J=7.1 Hz, 2H), 3.72 (t, 2H), 3.48 (q, 2H), 3.42 (t, 2H), 3.24 (s, 3H), 3.12 (t, 2H), 2.46 (s, 3H), 1.96 (p, J=6.2 Hz, 2H), 1.80 (p, 2H), 1.30 (t, J=7.1 Hz, 3H), 0.92 ( t, 2H), -0.11 (s, 9H).

Step C: ethyl 5-(3-iodopropyl)-2-[(3-methoxypropyl)(5-methyl-6-{[(2Z)-3-{[2(trimethylsilyl)ethoxy]methyl} -2,3- dihydro-1,3-benzothiazol-2-ylidene]amino}pyridazin-3-yl)amino]1,3-thiazol-4-carboxylate.

To a stirred solution of the product from step B (431 mg, 0.64 mmol, 1 eq.) in a 4:1 mixture of diethyl ether/acetonitrile (25 ml) was added imidazole (65.4 mg, 0.96 mmol, 1.5 eq.), triphenylphosphine (252 mg, 0.96 mmol, 1.5 eq.) and iodine (244 mg, 0.96 mmol, 1.5 eq.), and the mixture was stirred for 18 hours. The reaction mixture was diluted with ethyl acetate and washed successively with 10% aqueous sodium thiosulfate, water and brine, and dried ( magnesium sulfate) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 12 g RediSep™ silica gel cartridge) eluting with a gradient of 0-60% ethyl acetate in isoheptane gave the title product as a yellow gum (378 mg, 0.48 mmol, 75%).

LC/MS (C ₃₁ H ₄₃ IN ₆ O ₄ SiS ₂ ) 783 [M+H]+; RT 1.78 (ZHMS-U-V1).

^1H NMR (400 MHz, DMTO-sb,) δ 7.85 (d, 1H), 7.67 (s, 1H), 7.49-7.40 (m, 2H), 7.25 (ddd, J=8.2, 6.9, 1.6 Hz, 1H ), 5.87 (s, 2H), 4.38 (t, J=7.2 Hz, 2H), 4.29 (q, J=7.1 Hz, 2H), 3.72 (t, 2H), 3.42 (t, J=6.0 Hz, 2H ), 3.33 (t, 2H), 3.23 (s, 3H), 3.21-3.14 (m, 2H), 2.46 (s, 3H), 2.14 (p, J=6.9 Hz, 2H), 1.97 (p, J= 6.2 Hz, 2H), 1.32 (t, J=7.1

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Hz, 3H), 0.92 (t, 2H), -0.11 (s, 9H).

Step D: Ethyl 5-(3-{4-[3-(dimethylamino)prop-1-yn-1-yl]-2-fluorophenoxy}propyl)-2-[(3methoxypropyl)(5-methyl-6-{ [(2Z)-3-{[2-(trimethylsilyl)ethoxy]methyl}-2,3-dihydro-1,3-benzothiazol-2ylidene]amino}pyridazin-3-yl)amino]-1,3-thiazol- 4-carboxylate.

Sodium hydride (60% in mineral oil; 38.6 mg, 0.96 mmol, 2 eq.) was added to a solution of synthesis 6b (103 mg, 0.53 mmol, 1.1 eq.) in dimethylformamide (8 ml). After 10 min, the reaction mixture was cooled to 0°C and a solution of the product from step C (378 mg, 0.48 mmol, 1 eq.) in dimethylformamide (8 ml) was added. The mixture was allowed to warm to ambient temperature and stirred for 5 hours. The reaction mixture was quenched with water, acidified with 2N. aqueous hydrochloric acid solution and extracted with ethyl acetate (x2). The combined organic extracts were washed with brine, dried (magnesium sulfate) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 24 g RediSep™ silica gel cartridge) eluting with a gradient of 0-10% methanol in dichloromethane gave the title product as a brown foam (115 mg, 0.14 mmol, 28%).

LC/MS (C ₄₂ H ₅₄ FN ₇ O ₅ SiS ₂ ) 848 [M+H]+; RT 1.43 (LCMS-V-BI).

¹ H NMR (400 MHz, DMSOHO δ 7.84 (dd, J=7.5, 1.1 Hz, 1H), 7.67 (s, 1H), 7.49-7.41 (m, 2H), 7.39-7.34 (m, 1H), 7.28- 7.23 (m, 2H), 7.19 (t, J=8.6 Hz, 1H), 5.86 (s, 2H), 4.38 (t, J=7.3 Hz, 2H), 4.24 (q, J=7.1 Hz, 2H), 4.16 (t, J=6.1 Hz, 2H), 3.72 (t, J=8.0 Hz, 2H), 3.41 (t, J=5.9 Hz, 2H), 3.33 (s, 2H), 3.31-3.25 (m, 2H ), 3.24 (s, 3H), 2.35 (s, 2H), 2.48-2.41 (m, 9H), 2.13 (p, 2H), 1.97 (p, 2H), 1.29 (t, J=7.1 Hz, 3H), 0.92 (t, 2H), -0.12 (s, 9H).

Step E: ethyl 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(3methoxypropyl)amino)-5-(3 -{4—[3 - (dimethylamino)prop-1-yn-1-yl]-2-fluorophenoxy}propyl)-1,3-thiazol-4carboxylate.

Trifluoroacetic acid (1 ml, 13.4 mmol, 99 eq.) was added to a stirred solution of the product from step D (115 mg, 0.14 mmol, 1 eq.) in dichloromethane (3 ml), and the mixture was stirred for 18 hours. The reaction mixture was diluted dichloromethane, washed with saturated aqueous sodium bicarbonate, dried (magnesium sulfate) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 12 g RediSep™ silica gel cartridge) eluting with a gradient of 0-10% methanol in dichloromethane gave the title product as a yellow glass (32.2 mg, 0.04 mmol, 33%) .

LC/MS (C ₃₆ H ₄₀ FN ₇ O ₄ S ₂ ) 718 [M+H]+; RT 1.32 (LCMS-V-BI).

1H NMR (400 MHz, DMSO^) δ 7.91 (d, J=7.6 Hz, 1H), 7.67 (s, 1H), 7.54 (d, J=8.2 Hz, 1H), 7.44-7.37 (m, 1H), 7.31 (dd, 1H), 7.26-7.12 (m, 3H), 4.39 (t, J=7.2 Hz, 2H), 4.26 (q, J=7.0 Hz, 2H), 4.16 (t, J=6.1 Hz, 2H ), 3.43 (t, J=6.0 Hz, 2H), 3.40 (s, 2H), 3.31-3.26 (m, 2H), 3.25 (s, 3H), 2.47 (s, 3H), 2.21 (s, 6H) , 2.17-2.10 (m, 2H), 1.97 (p, 2H), 1.30 (t, J=7.1 Hz, 3H).

Step F: 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(3-methoxypropyl)amino)-5(3-{4-[3- (dimethylamino)prop-1-yn-1-yl]-2-fluorophenoxy}propyl)-1,3-thiazol-4-carboxylic acid.

To a solution of the product from step E (32.2 mg, 0.04 mmol, 1 eq.) in 1,4-dioxane (3 ml) was added lithium hydroxide monohydrate (9.41 mg, 0.22 mmol, 5 eq.) and the mixture was refluxed at during the night. The reaction mixture was allowed to cool to ambient temperature and concentrated in vacuo. The residue was triturated with water, then centrifuged, and the solvent was decanted. Toluene was added to the remaining solid and the mixture was sonicated, centrifuged and the solvent was decanted. Drying in vacuo provided the title product as a yellow solid (29.6 mg, 0.04 mmol, 96%) [as lithium salt].

HRMS-ERI (m/z) [M+H]+, calculated for C ₃₄ H ₃₇ FN ₇ O ₄ S ₂ : 690.2327; found: 690.2318.

Example 55: 2-[[6-[(4-Fluoro-1,3-benzothiazol-2-yl)amino]-4,5-dimethylpyridazin-3-yl]amino]thiazole4-carboxylic acid

Step A: N(6-chloro-4,5-dimethylpyridazin-3-yl)-4-fluoro-1,3-benzothiazol-2-amine.

To 158 mg 6-chloro-4,5-dimethylpyridazin-3-amine (1 mmol, 1 eq.) and 232 mg 2-bromo-4-fluoro-1,3benzothiazole (1 mmol, 1 eq.) in 8 ml 1 ,4-dioxane was added 160 mg of sodium hydride (60 wt.%, 4 mmol, 4 eq.), and the mixture was stirred at reflux for 1 hour. After cooling, the reaction mixture was quenched with water and the mixture was extracted with EtOAc. The combined and dried organic phases were concentrated to obtain 311 mg (101%) of the expected product.

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LC-MS-ESI (m/z): [M+H]+, calculated for C13H11C1FN4S: 309; found: 309.

Step B: ethyl 2-[[6-[(4-fluoro-1,3-benzothiazol-2-yl)amino]-4,5-dimethylpyridazin-3yl]amino]thiazol-4-carboxylate.

A mixture of 300 mg of the product from step A (0.97 mmol, 1 eq.), 167 mg of ethyl 2-aminothiazole-4-carboxylate (0.97 mmol, 1 eq.), 33 mg of Pd ₂ (dba) ₃ (0.036 mmol, 0.04 eq. ), 53 mg 4,5-bis(diphenylphosphino)-9,9dimethylxanthene (0.092 mmol, 0.1 eq.) and 0.23 ml DIPEA (2 eq.) in 8 ml 1,4-dioxane were stirred at 160°C in a microwave reactor in for 0.75 h. After cooling, the crude product was purified using reverse phase preparative chromatography to obtain 110 mg (26%) of the expected product.

1H NMR (400 MHz, DMSO-de) δ ppm 11.17 (s, 1H), 11.05 (s, 1H), 7.96 (s, 1H), 7.81 (m, 1H), 7.24 (m, 1H), 7.22 (m, 1H), 4.29 (q, 2H), 2.39 /2.37 (s+s, 6H), 1.31 (t, 3H).

Step C: 2-[[6-[(4-fluoro-1,3-benzothiazol-2-yl)amino]-4,5-dimethylpyridazin-3-yl]amino]thiazol-4carboxylic acid.

A mixture of the product from stage B, LiOHxH ₂ O, water and 1,4-dioxane was stirred at reflux for 9 hours. After cooling and concentration, 1N was added. HCl, and the precipitate was filtered to obtain the target product.

MSHR-ERI (m/z): [M+H] ⁺ , calculated for C ₁₇ H ₁₄ FN ₆ O ₂ S ₂ : 417.0604; found: 417.0595.

Example 56: 5-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)thiophene-2carboxylic acid

Step A: tert-butyl N[(tert-butoxy)carbonyl]-N-(6-chloro-4-methylpyridazin-3-yl)carbamate.

To a solution of di-tert-butyl dicarbonate (5.7 g, 26.1 mmol, 3 eq.) in dichloromethane (40 ml) was added 6-chloro-4-methylpyridazin-3-amine (1.25 g, 8.71 mmol, 1 eq.) followed by by adding 4dimethylaminopyridine (53.2 mg, 0.44 mmol, 0.05 eq.) and the mixture was stirred overnight. The reaction mixture was concentrated in vacuo and purified by flash column chromatography (50 g silica gel) eluting with 2:3 ethyl acetate/heptane to give a yellow solid. Trituration with heptane (15 ml) resulted in a solid, which was collected by filtration, washed with heptane (2 x 15 ml) and dried in vacuo to give the title product as a pale yellow solid (1.85 g, 5.38 mmol, 62%).

LC/MS (C ₁₅ H ₂₂ ClN ₃ O ₄ ) 188 [M-Boc-tBu+3H] ⁺ ; RT 1.28 (LCMS-V-BI).

1H NMR (400 MHz, CDCl3) δ 7.43 (d, 1H), 2.27 (d, 3H), 1.40 (s, 9H).

Step B: methyl 5-[(6-{bis[(tert-butoxy)carbonyl]amino}-5-methylpyridazin-3-yl)amino]thiophene 2-carboxylate.

Tris(dibenzylideneacetone)dipalladium(0) (107 mg, 0.12 mmol, 0.05 eq.) under nitrogen was added to a stirred solution of the product from step A (800 mg, 2.33 mmol, 1 eq.), methyl 5-aminothiophene-2-carboxylate (439 mg, 2.79 mmol, 1.2 eq.), X,X-diisopropylethylamine (1.16 ml, 6.98 mmol, 3 eq.) and xantphos (135 mg, 0.23 mmol, 0.1 eq.) in 1,4-dioxane (20 ml), and the mixture was heated at 100°C overnight. The reaction mixture was allowed to cool to ambient temperature and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 24 g RediSep™ silica gel cartridge) eluting with a gradient of 0-10% methanol in dichloromethane gave the material, which was triturated in diethyl ether and then the solids were collected by filtration to give the target product as a beige solid (442 mg, 0.95 mmol, 41%).

LC/MS (C ₂₁ H ₂₈ N ₄ O ₆ S) 465 [M+H] ⁺ ; RT 1.10 (LCMS-V-BI).

1H NMR (400 MHz, DMSO-de) δ 11.22 (s, 1H), 7.64 (d, J=4.2 Hz, 1H), 7.17 (s, 1H), 6.74 (d, J=4.3 Hz, 1H), 3.78 (s, 3H), 2.17 (s, 3H), 1.39 (s, 18H).

Step C: methyl 5-[(6-{bis[(tert-butoxy)carbonyl]amino}-5-methylpyridazin-3yl)(methyl)amino]thiophene-2-carboxylate.

Sodium hydride (60% dispersion; 60.9 mg, 1.52 mmol, 1.6 eq.) was added to a stirred solution of the product from step B (442 mg, 0.95 mmol, 1 eq.) in dimethylformamide (10 ml). After 15 min, iodomethane (0.12 mL, 1.9 mmol, 2 eq.) was added and the mixture was stirred at ambient temperature for 5 h. The reaction mixture was diluted with ethyl acetate, then washed with water (x2) followed by brine, dried (magnesium sulfate ) and concentrated in vacuo. Purification using automated flash column chromatography (CombiFlash Rf, 24 g RediSep™ silica gel cartridge) eluting with a gradient of 0-100% ethyl acetate in isoheptane resulted in

- 131 046336 to obtain the target product in the form of a creamy solid (295 mg, 0.62 mmol, 65%).

LC/MS (C ₂₂ H ₃₀ N ₄ O ₆ S) 479 [M+H]+; RT 1.23 (LCMS-V-BI).

1H NMR (400 MHz, DMSO) δ 7.73 (s, 1H), 7.69 (d, 1H), 6.92 (d, J=4.4 Hz, 1H), 3.79 (s, 3H), 3.66 (s, 3H), 2.22 (d, J=0.9 Hz, 3H), 1.40 (s, 18H).

Step D: Methyl 5-[(6-amino-5-methylpyridazin-3-yl)(methyl)amino]thiophene-2-carboxylate.

A solution of the product from step C (343 mg, 0.72 mmol, 1 eq.) in dichloromethane (8 ml) was cooled in an ice bath, then trifluoroacetic acid (2 ml, 26.8 mmol, 37 eq.) was added. The mixture was allowed to warm to ambient temperature and stirred for 18 hours. The solvent was removed in vacuo, then the residue was dissolved in methanol and loaded onto a methanol-conditioned SCX-2 cartridge (5 g). The cartridge was washed with methanol, then eluted with 3.5N. methanol ammonia. The solvent was removed in vacuo and the resulting solid was triturated with diethyl ether and filtered to give the title product as a cream solid (142 mg, 0.51 mmol, 71%).

LC/MS (C ₁₂ H ₁₄ N ₄ O ₂ S) 279 [M+H] ⁺ ; RT 0.67 (LCMS-V-BI).

1H NMR (400 MHz, DMSO-de) δ 7.59 (d, J=4.4 Hz, 1H), 7.35 (s, 1H), 6.58 (d, J=4.4 Hz, 1H), 6.00 (s, 2H), 3.75 (s, 3H), 3.50 (s, 3H), 2.16 (s, 3H).

Step E: methyl 5-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3yl} (methyl)amino)thiophene-2-car boxylate.

Tris(dibenzylideneacetone)dipalladium(0) (23.4 mg, 0.03 mmol, 0.05 eq.) under nitrogen was added to a stirred solution of the product from step D (142 mg, 0.51 mmol, 1 eq.), 2-bromo-1,3benzothiazole ( 131 mg, 0.61 mmol, 1.2 eq.), X,X-diisopropylethylamine (0.25 ml, 1.53 mmol, 3 eq.) and xantphos (29.5 mg, 0.05 mmol, 0.1 eq.) in 1,4-dioxane (5 ml) , and the mixture was heated at 100°C for 18 hours. The reaction mixture was allowed to cool to ambient temperature and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 12 g RediSep™ silica gel cartridge) eluting with a gradient of 0-6% methanol in dichloromethane provided the title product as a brown solid (158 mg, 0.38 mmol, 75%) .

LC/MS (C ₁₉ H ₁₇ N ₅ O ₂ S ₂ ) 412 [M+H] ⁺ ; RT 1.18 (LCMS-V-BI).

1H NMR (400 MHz, DMSO^) δ 11.04 (br s, 1H), 8.01 (br s, 1H), 7.70-7.63 (m, 3H), 7.43-7.35 (m, 1H), 7.27-7.17 (m, 1H), 6.80 (d, J=4.4 Hz, 1H), 3.81 (s, 3H), 3.63 (s, 3H), 2.46 (s, 3H).

Step F: 5-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)thiophene-2carboxylic acid.

To a suspension of the product from step E (158 mg, 0.38 mmol, 1 eq.) in 1,4-dioxane (10 ml) was added lithium hydroxide monohydrate (161 mg, 3.84 mmol, 10 eq.) and the mixture was refluxed at during the night. The reaction mixture was allowed to cool to ambient temperature and the resulting suspension was centrifuged, decanted and dried in vacuo to give the title product as a yellow solid (90.5 mg, 0.23 mmol, 59%).

MSVR-ERI (m/z) [MH] ^- , calculated for C ₁₈ H ₁₄ N ₅ O ₂ S ₂ : 396.0594; found: 396.0576.

Example 57: 2-[[6-[(5,6-Difluoro-1,3-benzothiazol-2-yl)amino]-4,5-dimethylpyridazin-3yl]amino]thiazol-4-carboxylic acid

Step A: N(6-chloro-4,5-dimethylpyridazin-3-yl)-5,6-difluoro-1,3-benzothiazol-2-amine.

To 158 mg of 6-chloro-4,5-dimethylpyridazin-3-amine (1 mmol, 1 eq.) and 250 mg of 2-bromo-5,6-difluoro-1,3-benzothiazole (1 mmol, 1 eq.) in 8 ml of 1,4-dioxane, 160 mg of sodium hydride (60 wt.%, 4 mmol, 4 eq.) was added and the mixture was stirred at reflux for 1 hour. After cooling, the reaction mixture was quenched with water and the mixture was extracted with EtOAc . The combined and dried organic phases were concentrated to obtain 300 mg (92%) of the expected product.

LC-MS-ESI (m/z): [M+H] ⁺ , calculated for C ₁₃ H ₁₀ ClF ₂ N ₄ S: 327; found: 327.

Step B: ethyl 2-[[6-[(5,6-difluoro-1,3-benzothiazol-2-yl)amino]-4,5-dimethylpyridazin-3yl]amino]thiazol-4-carboxylate.

A mixture of 300 mg of the product from step A (0.9 mmol, 1 eq.), 155 mg of ethyl 2-aminothiazole-4-carboxylate (0.9 mmol, 1 eq.), 33 mg of Pd ₂ (dba) ₃ (0.036 mmol, 0.04 eq. ), 53 mg of 4,5-bis(diphenylphosphino)-9,9dimethylxanthene (0.092 mmol, 0.1 equiv.) and 0.23 ml of DIPEA (2 equiv.) in 8 ml of 1,4-dioxane were stirred at 200°C in a microwave reactor in for 1 hour. After cooling, the crude product was purified using

- 132 046336 reverse-phase preparative chromatography to obtain 110 mg (26%) of the target product.

LC-MS-ESI (m/z): [M+H]+, calculated for C ₁₉ H ₁₇ F ₂ N ₆ O ₂ S ₂ : 463; found: 463.

Step C: 2-[[6-[(5,6-difluoro-1,3-benzothiazol-2-yl)amino]-4,5-dimethylpyridazin-3yl]amino]thiazol-4-carboxylic acid.

A mixture of the product from step B, LiOHxH ₂ O, water and 1,4-dioxane was stirred at reflux for 9 hours. After cooling, the crude product was purified by reverse phase preparative chromatography to obtain the desired product.

MSHR-ERI (m/z): [M+H]+, calculated for C ₁₇ H ₁₃ F ₂ N ₆ O ₂ S ₂ : 435.0509; found: 435.0506.

Example 58: 2-[[6-[(5-fluoro-1,3-benzothiazol-2-yl)amino]-4,5-dimethylpyridazin-3-yl]amino]thiazole4-carboxylic acid

Step A: N(6-chloro-4,5-dimethylpyridazin-3-yl)-5-fluoro-1,3-benzothiazol-2-amine.

To 158 mg of 6-chloro-4,5-dimethylpyridazin-3-amine (1 mmol, 1 eq.) and 348 mg of 2-bromo-5-fluoro-1,3-benzothiazole (1.5 mmol, 1.5 eq.) in 20 ml 1 ,4-dioxane, 160 mg sodium hydride (60 wt%, 4 mmol, 4 eq.) was added and the mixture was stirred at reflux for 1 hour. After cooling, the reaction mixture was quenched with 2 ml EtOH, concentrated and worked up 20 ml water. The precipitated solid was filtered to obtain 300 mg (97%) of the expected product.

¹ H NMR (500 MHz, DMSO-de) δ ppm 7.81 (dd, 1H), 7.29 (dd, 1H), 7.00 (td, 1H), 2.34 (s, 3H), 2.29 (s, 3H); ¹³ C NMR (125 MHz DMSOC) δ ppm 162.2, 161.9, 154.1, 152.0, 145.5, 137.4, 131.1, 125.8, 123.3, 110.6, 103.5, 15.7, 13.3; LC-MS-ESI (m/z): [M+H] ⁺ , calculated for C ₁₃ H ₁₁ ClFN ₄ S: 309.8; found: 309 and 307 [M-H] ^- .

Step B: ethyl 2-[[6-[(5-fluoro-1,3-benzothiazol-2-yl)amino]-4,5-dimethylpyridazin-3yl]amino]thiazol-4-carboxylate.

A mixture of 300 mg of the product from step A (0.97 mmol, 1 eq.), 167 mg of ethyl 2-aminothiazole-4-carboxylate (0.97 mmol, 1 eq.), 36 mg of Pd ₂ (dba) ₃ (0.039 mmol, 0.04 eq. ), 56 mg of 4,5-bis(diphenylphosphino)-9,9dimethylxanthene (0.097 mmol, 0.1 equiv.) and 0.34 ml of DIPEA (2 equiv.) in 10 ml of 1,4-dioxane were stirred at 200°C in a microwave reactor in for 1 hour. After cooling, the crude product was purified by reverse phase preparative chromatography to obtain 150 mg (35%) of the expected product.

LC-MS-ESI (m/z): [M+H] ⁺ , calculated for C ₁₉ H ₁₈ FN ₆ O ₂ S ₂ : 445.5; found: 445 and 443 [M-N] ^- .

Step C: 2-[[6-[(5-fluoro-1,3-benzothiazol-2-yl)amino]-4,5-dimethylpyridazin-3-yl]amino]thiazol-4carboxylic acid.

A mixture of 150 mg of the product from step B (0.338 mmol, 1 eq.), 28 mg of LiOHxH ₂ O (0.67 mmol, 2 eq.) and 10 drops of water in 10 ml of 1,4-dioxane was stirred under reflux for 3 h. After cooling and concentration, the crude product was purified using reverse phase preparative chromatography to obtain 112 mg (79%) of the expected product.

MSHR-ERI (m/z): [M+H] ⁺ , calculated for C ₁₇ H ₁₄ FN ₆ O ₂ S ₂ : 417.0604; found: 417.0598.

Example 59: 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)-5-[3-(2fluorophenoxy)propyl]-1 ,3-thiazole-4-carboxylic acid

Step A: ethyl 5-[3-(2-fluorophenoxy)propyl]-2-[methyl(5-methyl-6-{[(2Z)-3-{[2(trimethylsilyl)ethoxy]methyl}-2,3 -dihydro-1,3-benzothiazol-2-ylidene]amino}pyridazin-3-yl)amino]1,3-thiazol-4-carboxylate.

Sodium hydride (60% in mineral oil; 19.5 mg, 0.49 mmol, 2 eq.) was added to a solution of 2fluorophenol (0.03 mL, 0.29 mmol, 1.2 eq.) in dimethylformamide (4 mL). After 15 min, the reaction mixture was cooled to 0°C and a solution of synthesis product 5g (177 mg, 0.24 mmol, 1 eq.) in dimethylformamide (3 ml) was added. The mixture was allowed to warm to ambient temperature and stirred for 18 h.

- 133 046336

The reaction mixture was quenched by adding water, acidified with 2 N. aqueous hydrochloric acid solution and extracted with ethyl acetate (x2). The combined organic extracts were washed with brine, dried (magnesium sulfate) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 12 g RediSep™ silica gel cartridge) eluting with a gradient of 0-10% methanol in dichloromethane gave the title product as a beige foam (144 mg, 0.2 mmol, 83%).

LC/MS (C ₃₄ H ₄₁ FN ₆ O ₄ SiS ₂ ) 709 [M+H]+; RT 1.62 (LCMS-V-BI).

¹ H NMR (400 MHz, DMSO-de) δ 7.84 (dd, J=7.6, 1.0 Hz, 1H), 7.67 (d, J=1.1 Hz, 1H), 7.49-7.40 (m, 2H), 7.28-7.08 (m, 4H), 6.96-6.89 (m, 1H), 5.86 (s, 2H), 4.26 (q, J=7.1 Hz, 2H), 4.13 (t, J=6.2 Hz, 2H), 3.77 (s, 3H), 3.72 (t, 2H), 3.31-3.25 (m, 2H), 2.45 (d, J=1.0 Hz, 3H), 2.17-2.08 (m, 2H), 1.29 (t, J=7.1 Hz, 3H ), 0.92 (t, 2H), 0.12 (s, 9H).

Step B: ethyl 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)-5-[3-(2fluorophenoxy)propyl]- 1,3-thiazole-4-carboxylate.

Trifluoroacetic acid (1 mL, 13.4 mmol, 66 eq.) was added to a stirred solution of the product from Step A (144 mg, 0.2 mmol, 1 eq.) in dichloromethane (3 mL) and the mixture was stirred at ambient temperature overnight. The reaction mixture was partitioned between dichloromethane and saturated aqueous sodium bicarbonate, and the organic phase was washed with brine, dried (magnesium sulfate) and concentrated in vacuo. Purification by reverse phase automated flash chromatography (CombiFlash Rf, C18 13 g RediSep column) eluting with a gradient of 5-95% acetonitrile (pH 4; formic acid) in water (pH 4; formic acid) provided the desired product as a yellow solid (71 mg, 0.12 mmol, 61%).

LC/MS (C ₂₈ H ₂₇ FN ₆ O ₃ S ₂ ) 579 [M+H]+; RT 1.37 (LCMS-V-BI).

¹ H NMR (400 MHz, DMSO-de) δ 11.07 (br s, 1H), 7.92 (s, 1H), 7.69 (s, 1H), 7.61 (br s, 1H), 7.40 (t, J=7.6 Hz , 1H), 7.32-7.08 (m, 4H), 6.98-6.90 (m, 1H), 4.27 (q, J=7.1 Hz, 2H), 4.13 (t, J=6.2 Hz, 2H), 3.77 (s, 3H), 3.343.24 (m, 2H), 2.47 (s, 3H), 2.14 (p, J=6.2 Hz, 2H), 1.30 (t, J=7.1 Hz, 3H).

Step C: 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)-5-[3-(2fluorophenoxy)propyl] - 1 ,3-thiazole-4-carboxylic acid.

To a solution of the product from step B (71 mg, 0.12 mmol, 1 eq.) in 1,4-dioxane (4 ml) was added lithium hydroxide monohydrate (25.7 mg, 0.61 mmol, 5 eq.), and the mixture was refluxed at during the night. The reaction mixture was allowed to cool to ambient temperature and concentrated in vacuo. The residue was triturated with water and the solids were collected by filtration. The solid was further triturated with diethyl ether, filtered and dried in vacuo to give the title product as a yellow solid (49 mg, 0.09 mmol, 73%) [as lithium salt].

HRMS-ERI (m/z) [M+H]+, calculated for C ₂₆ H ₂₄ FN ₆ O ₃ S ₂ : 551.1330; found: 551.1335.

Example 60: 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)-5 -(3 methoxypropyl)-1,3-thiazole -4-carboxylic acid

Step A: ethyl 5-(3-methoxyprop-1-yn-1-yl)-2-[methyl(5-methyl-6-{[(2/)-3-{[2(trimethylsilyl)ethoxy]methyl} -2,3-dihydro-1,3-benzothiazol-2-ylidene]amino}pyridazin-3-yl)amino]1,3-thiazol-4-carboxylate.

Tetrakis(triphenylphosphine)palladium(0) (36.4 mg, 0.04 mmol, 0.1 eq.) under a nitrogen atmosphere was added to a stirred solution of synthesis product 11b (200 mg, 0.31 mmol, 1 eq.), methyl propargyl ether (0.06 ml, 0.76 mmol, 2.4 eq.), triethylamine (0.26 ml, 1.88 mmol, 6 eq.) and medip iodide (12 mg, 0.06 mmol, 0.2 eq.) in dimethylformamide (5 ml), and the mixture was heated at 100°C overnight. The reaction mixture was allowed to cool to ambient temperature and then concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 12 g RediSep™ silica gel cartridge) eluting with a gradient of 0-100% ethyl acetate in isoheptane provided the title product as a cream solid (78 mg, 0.12 mmol, 40%) .

LC/MS (C ₂₉ H ₃₆ N ₆ O ₄ SiS ₂ ) 625 [M+H]+; RT 1.56 (LCMS-V-BI).

1H NMR (400 MHz, DMSO-de) δ 7.86 (d, J=7.7 Hz, 1H), 7.74 (s, 1H), 7.51-7.41 (m, 2H), 7.26 (td, J=7.6, 1.5 Hz, 1H), 5.87 (s, 2H), 4.43 (s, 2H), 4.31 (q, J=7.1 Hz, 2H), 3.82 (s, 3H), 3.72 (t, J=8.0 Hz, 2H), 3.38 ( s, 3H), 2.47 (s, 3H), 1.32 (t, J=7.1 Hz, 3H), 0.91 (dd, J=10.7, 5.4 Hz, 2H), 0.00 (s, 9H).

Step B: ethyl 5-(3-methoxypropyl)-2-[methyl(5-methyl-6-{[(2/)-3-{[2- 134 046336 (trimethylsilyl)ethoxy]methyl}-2,3- dihydro-1,3-benzothiazol-2-ylidene]amino}pyridazin-3-yl)amino]1,3-thiazol-4-carboxylate.

A suspension of the product from step A (78 mg, 0.12 mmol, 1 eq.) in a 1:1 mixture of ethyl acetate/methanol (8 ml) was loaded into a flask containing platinum oxide PU (2.83 mg, 0.01 mmol, 0.1 eq.) under a nitrogen atmosphere . The vessel was evacuated and filled again with nitrogen (N3), then evacuated, created a hydrogen atmosphere and shaken overnight. The mixture was filtered through celite (2.5 g), eluted with methanol, and the solvent was removed in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 12 g RediSep™ silica gel cartridge) eluting with a gradient of 0-10% methanol in dichloromethane gave the title product as a white foam (56.1 mg, 0.09 mmol, 71%).

LC/MS (C ₂₉ H ₄₀ N ₆ O ₄ SiS ₂ ) 629 [M+H]+; RT 1.54 (LCMS-V-BI).

¹ H NMR (400 MHz, DMSO^) δ 7.84 (dd, J=7.7, 1.0 Hz, 1H), 7.66 (d, J=1.1 Hz, 1H), 7.49-7.41 (m, 2H), 7.25 (ddd, J=8.4, 6.9, 1.6 Hz, 1H), 5.86 (s, 2H), 4.28 (q, J=7.1 Hz, 2H), 3.77 (s, 3H), 3.72 (t, 2H), 3.38 (t, J =6.3 Hz, 2H), 3.26 (s, 3H), 3.189-3.09 (m, 2H), 2.46 (d, J=1.1 Hz, 3H), 1.86 (p, 1H), 1.31 (t, J=7.1 Hz , 3H), 0.97-0.87 (m, 2H), 0.00 (s, 9H).

Step C: ethyl 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)-5-(3methoxypropyl)-1,3-thiazole -4-carboxylate.

Trifluoroacetic acid (1 mL, 13.4 mmol, 150 eq.) was added to a stirred solution of the product from Step B (56 mg, 0.09 mmol, 1 eq.) in dichloromethane (3 mL), and the mixture was stirred at ambient temperature overnight. The reaction mixture was partitioned between dichloromethane and saturated aqueous sodium bicarbonate, and the organic phase was washed with brine, dried (magnesium sulfate) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 12 g RediSep™ silica gel cartridge) eluting with a gradient of 0-6% methanol in dichloromethane provided the title product as a yellow glass (40 mg, 0.08 mmol, 90%) .

LC/MS (C2 ₃ H2 ₆ N ₆ O ₃ S2) 499 [M+H]+; RT 1.25 (LCMS-V-BI).

1H NMR (400 MHz, DMSO^) δ 11.11 (br s, 1H), 7.93 (br s, 1H), 7.69 (s, 1H), 7.58 (br s, 1H), 7.39 (t, J=7.5 Hz, 1H), 7.22 (t, 1H), 4.29 (q, J=7.1 Hz, 2H), 3.77 (s, 3H), 3.39 (t, J=6.3 Hz, 2H), 3.27 (s, 3H), 3.18- 3.10 (m, 2H), 2.47 (s, 3H), 1.94-1.82 (m, 2H), 1.33 (t, J=7.1 Hz, 3H).

Step D: 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl} (methyl)amino)-5-(3 methoxypropyl)-1,3-thiazole -4-carboxylic acid.

To a solution of the product from step C (40 mg, 0.08 mmol, 1 eq.) in 1,4-dioxane (3 ml) was added lithium hydroxide monohydrate (16.8 mg, 0.4 mmol, 5 eq.) and the mixture was refluxed at for 5 hours. The reaction mixture was allowed to cool to ambient temperature, concentrated in vacuo, triturated with water and the solids collected by filtration. The solid was suspended in 1,4-dioxane (4 ml) and then hydrochloric acid (4 M in 1,4-dioxane; 0.5 ml) was added. The suspension was stirred at ambient temperature for 2 hours, then centrifuged and the solvent was removed by decanting. The solid was washed with 1,4-dioxane (2x2 ml), then centrifuged again and the solvent was removed by decanting. The solid was finally triturated with water, filtered and dried in vacuo to give the title product as an off-white solid (28.4 mg, 0.06 mmol, 75%) [as hydrochloric acid salt].

HRMS-ERI (m/z) [M+H]+, calculated for C ₂ iH ₂ 3N6O3S ₂ : 471.1268; found: 471.1271.

Example 61: 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)-5-[3(cyclohexyloxy)propyl]-1, 3-thiazole-4-carboxylic acid

Step A: ethyl 5-[3-(cyclohexyloxy)prop-1-yn-1-yl]-2-[methyl(5-methyl-6-{[(2Z)-3-{[2(trimethylsilyl)ethoxy] methyl}-2,3-dihydro-1,3-benzothiazol-2-ylidene] amino} pyridazin-3-yl)amino] 1,3-thiazol-4-carboxylate.

Tetrakis(triphenylphosphine)palladium(0) (36.4 mg, 0.04 mmol, 0.1 equiv.) under nitrogen atmosphere was added to a stirred solution of synthesis product 11b (200 mg, 0.31 mmol, 1 equiv.), (prop-2-yn-1yloxy) cyclohexane (120 mg, 0.86 mmol, 2.76 eq.), triethylamine (0.26 ml, 1.88 mmol, 6 eq.) and medip iodide (12 mg, 0.06 mmol, 0.2 eq.) in dimethylformamide (6 ml), and the mixture was heated at 100°C overnight. The reaction mixture was allowed to cool to ambient temperature, then diluted with ethyl acetate, washed with water (x2) followed by brine, dried (sulfate

- 135 046336 magnesium) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 12 g RediSep™ silica gel cartridge) eluting with a gradient of 0-70% ethyl acetate in isoheptane provided the title product as a cream solid (50 mg, 0.07 mmol, 23%).

LC/MS (C ₃₄ H ₄₄ N ₆ O ₄ SiS ₂ ) 693 [M+H]+; RT 1.66 (LCMS-V-BI).

1H NMR (400 MHz, DMSO^) δ 7.85 (d, J=7.7 Hz, 1H), 7.75 (d, J=1.1 Hz, 1H), 7.52-7.39 (m, 2H), 7.25 (td, J=7.5 , 1.4 Hz, 1H), 5.88 (s, 2H), 4.48 (s, 2H), 4.30 (q, J=7.1 Hz, 2H), 3.82 (s, 3H), 3.72 (t, 2H), 3.57-3.49 (m, 1H), 2.48 (d, J=1.0 Hz, 3H), 1.95-1.86 (m, 2H), 1.73-1.65 (m, 2H), 1.55-1.46 (m, 2H), 1.32 (t, 3H ), 1.321.22 (m, 4H), 0.92 (dd, J=8.5, 7.5 Hz, 2H), 0.00 (s, 9H).

Step B: ethyl 5-[3-(cyclohexyloxy)propyl]-2-[methyl(5-methyl-6-{[(2Z)-3-{[2(trimethylsilyl)ethoxy]methyl}-2,3-dihydro -1,3-benzothiazol-2-ylidene]amino}pyridazin-3-yl)amino]1,3-thiazol-4-carboxylate.

A suspension of the product from step A (50 mg, 0.07 mmol, 1 eq.) in a 1:1 mixture of ethyl acetate/methanol (6 ml) was loaded into a flask containing platinum oxide (21.6 mg, 0.01 mmol, 0.1 eq.) in nitrogen atmosphere. The vessel was evacuated and again filled with nitrogen (x3), then evacuated, created an atmosphere of hydrogen and shaken for 18 hours. The reaction mixture was filtered through celite (2.5 g), eluted with ethyl acetate and methanol, and the solvent was removed in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 12 g RediSep™ silica gel cartridge) eluting with a gradient of 0-60% ethyl acetate in isoheptane provided the title product as a colorless glass (42.2 mg, 0.06 mmol, 84%) .

LC/MS (C ₃₄ H ₄₈ N ₆ O ₄ SiS ₂ ) 697 [M+H] ⁺ ; RT 1.71 (LCMS-V-BI).

¹ H NMR (400 MHz, DMSO^) δ 7.82 (d, J=7.5 Hz, 1H), 7.67 (d, J=1.1 Hz, 1H), 7.50-7.41 (m, 2H), 7.287.23 (m, 1H), 5.87 (s, 2H), 4.28 (q, J=7.1 Hz, 2H), 3.77 (s, 3H), 3.72 (t, 2H), 3.45 (t, J=6.1 Hz, 2H), 3.27- 3.20 (m, 1H), 3.18-3.11 (m, 2H), 2.46 (d, J=1.0 Hz, 3H), 1.90-1.79 (m, 4H), 1.72-1.62 (m, 2H), 1.51-1.42 ( m, 2H), 1.31 (t, J=7.1 Hz, 3H), 1.28-1.17 (m, 5H), 0.96-0.89 (m, 2H), 0.00 (s, 9H).

Step C: ethyl 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)-5-[3(cyclohexyloxy)propyl]-1 ,3-thiazole-4-carboxylate.

Trifluoroacetic acid (0.6 mL, 8.06 mmol, 134 eq.) was added to a stirred solution of the product from Step B (42 mg, 0.06 mmol, 1 eq.) in dichloromethane (3 mL), and the mixture was stirred at ambient temperature overnight. The reaction mixture was partitioned between dichloromethane and saturated aqueous sodium bicarbonate, and the organic phase was washed with brine, dried (magnesium sulfate) and concentrated in vacuo. Purification by reverse phase automated flash chromatography (CombiFlash Rf, C18 13 g RediSep column) eluting with a gradient of 5-95% acetonitrile (pH 4; formic acid) in water (pH 4; formic acid) provided the desired product in the form of a yellow glassy substance (19.6 mg, 0.03 mmol, 57%).

LC/MS (C ₂₈ H ₃₄ N ₆ O ₃ S ₂ ) 567 [M+H] ⁺ ; RT 1.43 (LCMS-V-BI).

1H NMR (400 MHz, DMSO) δ 11.085 (br s, 1H), 7.90 (s, 1H), 7.68 (s, 1H), 7.60 (br s, 1H), 7.38 (t, 1H), 7.22 (t, 1H), 4.29 (q, J=7.1 Hz, 2H), 3.77 (s, 3H), 3.46 (t, J=6.2 Hz, 2H), 3.28-3.20 (m, 1H), 3.15 (t, 2H), 2.47 (s, 3H), 1.92-1.81 (m, 4H), 1.73-1.62 (m, 2H), 1.52-1.43 (m, 2H), 1.32 (t, 3H), 1.29-1.15 (m, 4H).

Step D: 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)-5-[3(cyclohexyloxy)propyl]-1, 3-thiazole-4-carboxylic acid.

To a solution of the product from step C (19.6 mg, 0.03 mmol, 1 eq.) in 1,4-dioxane (4 ml) was added lithium hydroxide monohydrate (7.26 mg, 0.17 mmol, 5 eq.), and the mixture was heated at 100°C for 6.5 h, then allowed to cool to ambient temperature and concentrated in vacuo. The residue was triturated with water, filtered and dried in vacuo to give the title product as a yellow solid (11 mg, 0.02 mmol, 59%) [as lithium salt].

MSHR-ERI (m/z) [M+H] ⁺ , calculated for C26H31N6O3S2: 539.1894; found: 539.1895.

Example 62: 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-cyclopropylpyridazin-3-yl}(methyl)amino)-5(3 -{4-[3 -(dimethylamino )prop-1 -yn-1 -yl]-2-fluorophenoxy}propyl)-1,3-thiazol-4-carboxylic acid

Step A: ethyl 2-[(5-cyclopropyl-6-{[(2Z)-3-{[2-(trimethylsilyl)ethoxy]methyl}-2,3-dihydro-1,3

- 136 046336 benzothiazol-2-ylidene]amino}pyridazin-3 -yl)(methyl)amino]-5 -(3-hydroxyprop-1-in-1-yl)-1,3-thiazol-4carboxylate.

Tetrakis(triphenylphosphine)palladium(0) (422 mg, 0.36 mmol, 0.1 eq.) under a nitrogen atmosphere was added to a stirred solution of synthesis product 11d (2.42 g, 3.65 mmol, 1 eq.), propargyl alcohol (0.86 ml, 14.6 mmol, 4 eq.), triethylamine (3.04 ml, 21.9 mmol, 6 eq.) and copper iodide (139 mg, 0.72 mmol, 0.2 eq.) in dimethylformamide (50 ml), and the mixture was heated at 100°C overnight. The reaction mixture was allowed to cool to ambient temperature and then concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 80 g RediSep™ silica gel cartridge) eluting with a gradient of 0-100% ethyl acetate in isoheptane provided the title product as a cream solid (0.91 g, 1.43 mmol, 39%) .

LC/MS (CrH.’vNfO.-iSiSj 637 [M+H]+; RT 1.44 (LCMS-V-BI).

1H NMR (400 MHz, DMSO^ ₆ ) δ 11.18 (br s, 1H), 7.87 (d, 1H), 7.50-7.40 (m, 2H), 7.26 (td, 1H), 7.11 (s, 1H), 6.00 (d, J=10.2 Hz, 2H), 5.88 (s, 2H), 5.43 (t, 1H), 4.39 (d, J=5.9, 0.8 Hz, 2H), 4.30 (q, J=7.1 Hz, 2H) , 3.82 (s, 3H), 3.75-3.67 (m, 2H), 2.72-2.62 (m, 1H), 1.32 (t, J=7.1 Hz, 3H), 1.22-1.12 (m, 2H), 1.12-1.06 (m, 2H), 0.94-0.85 (m, 2H), 0.00 (s, 9H).

Step B: ethyl 2-[(5-cyclopropyl-6-{[(22)-3-{[2-(trimethylsilyl)ethoxy]methyl}-2,3-dihydro-1,3benzothiazol-2-ylidene] amino} pyridazin-3-yl)(methyl)amino]-5-(3-hydroxypropyl)-1,3-thiazol-4carboxylate.

A suspension of the product from step A (913 mg, 1.43 mmol, 1 eq.) in a 1:1 mixture of ethyl acetate/methanol (40 ml) was loaded into a flask containing platinum oxide (32.6 mg, 0.14 mmol, 0.1 eq.) under a nitrogen atmosphere . The vessel was evacuated and again filled with nitrogen (x3), then evacuated, created an atmosphere of hydrogen and shaken for 3.5 hours. The reaction mixture was filtered through celite (10 g), eluted with methanol, and the solvent was removed in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 12 g RediSep™ silica gel cartridge) eluting with a gradient of 0-70% ethyl acetate in isoheptane gave the title product as a creamy foam (762 mg, 1.19 mmol, 83%).

LC/MS (C30H40N6O4SiS2) 641 [M+H]+; RT 1.44 (LCMS-V-BI).

1H NMR (400 MHz, DMSO-de) δ 7.85 (dd, J=7.5, 1.0 Hz, 1H), 7.49-7.39 (m, 2H), 7.28-7.22 (m, 1H), 7.07 (s, 1H), 5.87 (s, 2H), 4.56 (t, J=5.1, 3.1 Hz, 1H), 4.28 (q, J=7.1 Hz, 3H), 3.76 (s, 3H), 3.75-3.66 (m, 2H), 3.48 (q, 2H), 3.17-3.07 (m, 2H), 2.71-2.62 (m, 1H), 1.79 (p, 2H), 1.31 (t, J=7.1 Hz, 3H), 1.19-1.12 (m, 2H ), 1.11-1.04 (m, 2H), 0.94-0.86 (m, 2H), 0.00 (s, 9H).

Step C: ethyl 2-[(5-cyclopropyl-6-{[(22)-3-{[2-(trimethylsilyl)ethoxy]methyl}-2,3-dihydro-1,3benzothiazol-2-ylidene]amino} pyridazin-3-yl)(methyl)amino]-5-(3-iodopropyl)-1,3-thiazol-4-carboxylate.

To a stirred solution of the product from step B (762 mg, 1.19 mmol, 1 eq.) in a 10:3 mixture of diethyl ether/acetonitrile (26 ml) was added imidazole (122 mg, 1.78 mmol, 1.5 eq.), triphenylphosphine (468 mg, 1.78 mmol, 1.5 eq.) and iodine (453 mg, 1.78 mmol, 1.5 eq.), and the mixture was stirred at ambient temperature overnight. The reaction mixture was diluted with ethyl acetate, washed with 10% aqueous sodium thiosulfate followed by water, then brine, dried (magnesium sulfate) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 40 g RediSep™ silica gel cartridge) eluting with a gradient of 0-60% ethyl acetate in isoheptane gave the title product as a creamy foam (613 mg, 0.82 mmol, 69%).

LC/MS (C ₃₀ H ₃ 9IN ₆ O ₃ SiS ₂ ) 751 [M+H]+; RT 1.62 (LCMS-V-BI).

1H NMR (400 MHz, DMSOO δ 7.84 (dd, J=7.7, 1.0 Hz, 1H), 7.49-7.38 (m, 2H), 7.25 (ddd, J=8.2, 7.0, 1.5 Hz, 1H), 7.07 (s , 1H), 5.88 (s, 2H), 4.30 (q, J=7.1 Hz, 2H), 3.75 (s, 3H), 3.74-3.65 (m, 2H), 3.38-3.29 (m, 2H), 3.17 ( t, J=6.8 Hz, 2H), 2.70-2.60 (m, 1H), 2.14 (p, 2H), 1.32 (t, J=7.1 Hz, 3H), 1.20-1.11 (m, 2H), 1.10-1.04 (m, 2H), 0.95-0.85 (m, 2H), 0.00 (s, 9H).

Step D: Ethyl 2-[(5-cyclopropyl-6-{[(22)-3-{[2-(trimethylsilyl)ethoxy]methyl}-2,3-dihydro-1,3benzothiazol-2-ylidene]amino} pyridazin-3 -yl)(methyl)amino]-5 -(3-{4-[3 -(dimethylamino)prop-1 -in-1 -yl] 2-fluorophenoxy}propyl)-1,3-thiazol-4 -carboxylate.

Sodium hydride (60% in mineral oil; 65.3 mg, 1.63 mmol, 2 eq.) was added to a solution of synthesis 6b (189 mg, 0.98 mmol, 1.2 eq.) in dimethylformamide (10 ml). After 10 min, the reaction mixture was cooled to 0°C and a solution of the product from step C (613 mg, 0.82 mmol, 1 eq.) in dimethylformamide (10 ml) was added. The mixture was allowed to warm to ambient temperature and stirred for 3.5 hours. The reaction mixture was quenched by adding water, acidified with 2N. aqueous solution of hydrochloric acid and extracted with dichloromethane (x2). The combined organic extracts were washed with brine, dried (magnesium sulfate) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 24 g RediSep™ silica gel cartridge) eluting with a gradient of 0-10% methanol in dichloromethane resulted in a solid which was further purified by preparative HPLC (HPLC-V-AI method) yielding the desired product as an off-white solid (283 mg, 0.35 mmol, 43%).

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LC/MS (C ₄₁ H ₅₀ FN ₇ O ₄ SiS2) 816 [M+H]+; RT 1.41 (LCMS-V-BI).

1H NMR (400 MHz, DMSO-de) δ 8.35 (s, 1H), 7.82 (d, 1H), 7.48-7.39 (m, 2H), 7.33-7.11 (m, 3H), 7.06 (s, 1H), 5.86 (s, 2H), 4.25 (q, J=7.1 Hz, 2H), 4.13 (t, J=6.2 Hz, 2H), 3.75 (s, 3H), 3.70 (t, 2H), 3.37 (s, 2H ), 3.25 (t, J=7.6 Hz, 2H), 2.69-2.59 (m, 1H), 2.19 (s, 6H), 2.10 (p, J=6.3 Hz, 2H), 1.28 (t, J=7.1 Hz , 3H), 1.19-1.11 (m, 2H), 1.10-1.02 (m, 2H), 0.89 (dd, J=8.7, 7.4 Hz, 2H), 0.00 (s, 9H).

Step E: ethyl 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-cyclopropylpyridazin-3-yl}(methyl)amino)5-(3-{4-[3-( dimethylamino)prop-1-yn-1-yl]-2-fluorophenoxy}propyl)-1,3-thiazol-4-carboxylate.

Trifluoroacetic acid (4 mL, 53.7 mmol, 155 eq.) was added to a stirred solution of the product from Step D (283 mg, 0.35 mmol, 1 eq.) in dichloromethane (10 mL), and the mixture was stirred at ambient temperature overnight. The reaction mixture was partitioned between dichloromethane and saturated aqueous sodium bicarbonate, washed with brine, dried (magnesium sulfate) and concentrated in vacuo. Purification by reverse phase automated flash chromatography (CombiFlash Rf, C18 13 g RediSep column) eluting with a gradient of 5-95% acetonitrile (pH 4; formic acid) in water (pH 4; formic acid) gave the desired product in the form of a yellow glassy substance (56.5 mg, 0.08 mmol, 24%).

LC/MS (C ₃₅ H ₃₆ FN ₇ O ₃ S ₂ ) 686 [M+H]+; RT 1.12 (LCMS-V-BI).

¹ H NMR (400 MHz, DMSS)-с1 ₆ ) δ 11.71 (br s, 1H), 7.89 (d, 1H), 7.50 (d, J=8.0 Hz, 1H), 7.39 (ddd, J=8.2, 7.3 , 1.3 Hz, 1H), 7.31 (dd, J=12.0, 2.0 Hz, 1H), 7.27-7.10 (m, 4H), 4.26 (q, J=7.1 Hz, 2H), 4.15 (t, J=6.2 Hz , 2H), 3.77 (s, 3H), 3.40 (s, 2H), 3.30-3.24 (m, 2H), 2.49-2.45 (m, 1H), 2.21 (s, 6H), 2.13 (p, J=6.3 Hz, 2H), 1.30 (t, J=7.1 Hz, 2H), 1.20-1.10 (m, 2H), 1.06-0.99 (m, 2H).

Step F: 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-cyclopropylpyridazin-3-yl}(methyl)amino)-5-(3{4-[3-(dimethylamino )prop-1-yn-1-yl]-2-fluorophenoxy}propyl)-1,3-thiazol-4-carboxylic acid.

To a solution of the product from step E (56.5 mg, 0.08 mmol, 1 eq.) in 1,4-dioxane (4 ml) was added lithium hydroxide monohydrate (17.3 mg, 0.41 mmol, 5 eq.) and the mixture was heated at 100°C for 6.5 hours. The reaction mixture was allowed to cool to ambient temperature and concentrated in vacuo. The residue was triturated with water, filtered and dried in vacuo to give the title product as a yellow solid (46.5 mg, 0.07 mmol, 86%) [as lithium salt].

HRMS-ERI (m/z) [M+H]+, calculated for C ₃₃ H ₃₃ FN ₇ O ₃ S ₂ : 658.2065; found: 658.2070.

Example 63: 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)-5-(3phenoxypropyl)-1,3-thiazol- 4-carboxylic acid

Step A: ethyl 2-[methyl(5-methyl-6-{[(2Z)-3-{[2-(trimethylsilyl)ethoxy]methyl}-2,3-dihydro-1,3benzothiazol-2-ylidene] amino }pyridazin-3 -yl)amino]-5-(3-phenoxyprop-1 -yn-1 -yl)-1,3-thiazol-4carboxylate.

Tetrakis(triphenylphosphine)palladium(0) (85.2 mg, 0.08 mmol, 0.1 eq.) was added under nitrogen atmosphere to a stirred solution of synthesis product 11b (469 mg, 0.74 mmol, 1 eq.), phenylpropargyl ether (0.38 ml, 2.96 mmol, 4 eq.), triethylamine (0.62 ml, 4.42 mmol, 6 eq.) and copper(G) iodide (28.1 mg, 0.14 mmol, 0.2 eq.) in dimethylformamide (12 ml), and the mixture was heated at 100°C for nights. The reaction mixture was allowed to cool to ambient temperature and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 40 g RediSep™ silica gel cartridge) eluting with a gradient of 0-100% ethyl acetate in isoheptane gave the title product as a brown foam (296 mg, 0.43 mmol, 58%).

LC/MS (C34H38N6O ₄ SiS2) 687 [M+H]+; RT 1.57 (LCMS-V-BI).

1H NMR (400 MHz, DMSO^6) δ 7.87 (dd, J=7.8, 1.0 Hz, 1H), 7.74 (d, J=1.1 Hz, 1H), 7.51-7.42 (m, 2H), 7.38-7.33 ( m, 2H), 7.26 (ddd, J=9.2, 5.5, 1.5 Hz, 1H), 7.10-7.06 (m, 2H), 7.00 (tt, J=7.4, 1.1 Hz, 1H), 5.87 (s, 2H) , 5.15 (s, 2H), 4.26 (q, J=7.1 Hz, 2H), 3.83 (s, 3H), 3.76-3.70 (m, 2H), 2.47 (d, J=1.0 Hz, 3H), 1.28 ( t, J=7.1 Hz, 3H), 0.97-0.88 (m, 2H), 0.00 (s, 9H).

Step B: ethyl 2-[methyl(5-methyl-6-{[(2Z)-3-{[2-(trimethylsilyl)ethoxy]methyl}-2,3-dihydro-1,3benzothiazol-2-ylidene]amino }pyridazin-3-yl)amino]-5-(3-phenoxypropyl)-1,3-thiazol-4-carboxylate.

A suspension of the product from step A (296 mg, 0.43 mmol, 1 eq.) in a 1:1 mixture of ethyl acetate/methanol (10 ml) was loaded into a flask containing platinum oxide (PO) (9.78 mg, 0.04 mmol, 0.1 eq.) in nitrogen atmosphere. The vessel was evacuated and refilled with nitrogen (x3), then evacuated, created a hydrogen atmosphere and shaken overnight. The reaction mixture was filtered through celite (2.5 g), eluted with methanol, and the solvent was removed in vacuo. Cleaning with automatic flash column

- 138 046336 chromatography (CombiFlash Rf, 12 g RediSep™ silica gel cartridge) eluting with a gradient of 0-40% ethyl acetate in isoheptane gave the desired product as a brown gum (148 mg, 0.21 mmol, 50%).

LC/MS (C ₃₄ H ₄₂ N ₆ O ₄ SiS ₂ ) 691 [M+H]+; RT 1.59 (LCMS-V-BI).

¹ H NMR (400 MHz, DMSO^ ₆ ) δ 7.84 (d, J=7.7 Hz, 1H), 7.67 (d, J=1.2 Hz, 1H), 7.51-7.40 (m, 2H), 7.337.22 (m , 3H), 7.05-6.88 (m, 3H), 5.76 (s, 2H), 4.26 (q, 2H), 4.04 (t, J=6.2 Hz, 2H), 3.78 (s, H), 3.73 (t, 2H), 3.333.22 (m, 2H), 2.46 (s, 3H), 2.11 (p, 1H), 1.30 (t, J=7.1 Hz, 3H), 0.93 (t, J=8.0 Hz, 2H), 0.00 (s, 9H).

Step C: ethyl 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)-5-(3phenoxypropyl)-1,3-thiazole -4-carboxylate.

Hydrochloric acid (4 M in 1,4-dioxane; 3 mL) was added slowly to a solution of the product from Step B (148 mg, 0.21 mmol, 1 eq.) in tetrahydrofuran (5 mL), and the mixture was stirred at ambient temperature for nights. The reaction mixture was partitioned between ethyl acetate and saturated aqueous sodium bicarbonate, washed with brine, dried (magnesium sulfate) and concentrated in vacuo. Purification by reverse phase automated flash chromatography (CombiFlash Rf, C18 13 g RediSep column) eluting with a gradient of 5-95% acetonitrile (pH 4; formic acid) in water (pH 4; formic acid) gave the desired product as yellow glassy substance (46.7 mg, 0.08 mmol, 39%).

LC/MS (C ₂₈ H ₂₈ N ₆ O ₃ S ₂ ) 561 [M+H]+; RT 1.37 (LCMS-V-BI).

^1H NMR (400 MHz, DMSO^ ₆ ) δ 11.07 (br s, 1H), 8.01-7.81 (m, 2H), 7.69 (s, 1H), 7.39 (t, 1H), 7.35-7.26 (m, 2H ), 7.25-7.19 (m, 1H), 7.00-6.88 (m, 3H), 4.28 (q, J=7.1 Hz, 2H), 4.05 (t, J=6.2 Hz, 2H), 3.78 (s, 3H) , 3.343.24 (m, 2H), 2.47 (s, 3H), 2.21-1.99 (m, 2H), 1.31 (t, J=7.1 Hz, 3H).

Step D: 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)-5-(3phenoxypropyl)-1,3-thiazol- 4-carboxylic acid.

To a solution of the product from step C (46.7 mg, 0.08 mmol, 1 eq.) in 1,4-dioxane (4 ml) was added lithium hydroxide monohydrate (17.5 mg, 0.42 mmol, 5 eq.), and the mixture was heated at 100°C for 6 hours. The reaction mixture was allowed to cool to ambient temperature and concentrated in vacuo. The residue was triturated with water, filtered and dried in vacuo to give the title product as a yellow solid (28.7 mg, 0.05 mmol, 65%) [as lithium salt].

MSVR-ERI (m/z) [MH] ^- , calculated for C ₂₆ H ₂₃ N ₆ O ₃ S ₂ : 531.1279; found: 531.1285.

Example 64: 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(3-methoxypropyl)amino)5-(3-methoxypropyl)-1,3 -thiazole-4-carboxylic acid

Step A: ethyl 5-(3-methoxyprop-1-yn-1-yl)-2-[(3-methoxypropyl)(5-methyl-6-{[(27)-3-{[2(trimethylsilyl)ethoxy ]methyl}-2,3-dihydro-1,3-benzothiazol-2-ylidene]amino}pyridazin-3-yl)amino]1,3-thiazol-4-carboxylate.

Tetrakis(triphenylphosphine)palladium(0) (83.3 mg, 0.08 mmol, 0.1 equiv.) was added under a nitrogen atmosphere to a stirred solution of synthesis product 11c (500 mg, 0.72 mmol, 1 equiv.), methyl propargyl ether (0.24 ml, 2.88 mmol, 4 equiv.), triethylamine (0.6 ml, 4.32 mmol, 6 equiv.) and copper(G) iodide (27.5 mg, 0.14 mmol, 0.2 equiv.) in dimethylformamide (10 ml), and the mixture was heated at 100°C for nights. The reaction mixture was allowed to cool to ambient temperature and then concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 40 g RediSep™ silica gel cartridge) eluting with a gradient of 0-80% ethyl acetate in isoheptane gave the title product as a brown gum (337 mg, 0.49 mmol, 68%).

LC/MS (C ₃₂ H ₄₂ N ₆ O ₅ SiS ₂ ) 683 [M+H]+; RT 1.57 (LCMS-V-BI).

1H NMR (400 MHz, DMSO-ds) δ 7.87 (dd, J=7.4, 1.1 Hz, 1H), 7.73 (d, J=1.1 Hz, 1H), 7.51-7.42 (m, 2H), 7.26 (dt, 1H), 5.87 (s, 2H), 4.51-4.44 (m, 2H), 4.43 (s, 2H), 4.30 (q, J=7.1 Hz, 2H), 3.72 (t, 2H), 3.42 (t, J =5.9 Hz, 2H), 3.37 (s, 3H), 3.23 (s, 3H), 2.48 (d, J=0.9 Hz, 3H), 1.99 (p, J=6.6 Hz, 2H), 1.32 (t, J =7.1 Hz, 3H), 0.92 (t, J=8.6, 7.4 Hz, 2H), 0.00 (s, 9H).

Step B: ethyl 5-(3-methoxypropyl)-2-[(3-methoxypropyl)(5-methyl-6-{[(27)-3-{[2(trimethylsilyl)ethoxy]methyl}-2,3- dihydro-1,3-benzothiazol-2-ylidene]amino}pyridazin-3-yl)amino]1,3-thiazol-4-carboxylate.

- 139 046336

A suspension of the product from step A (337 mg, 0.49 mmol, 1 eq.) in a 1:1 mixture of ethyl acetate/methanol (10 ml) was loaded into a flask containing platinum(^) oxide (11.2 mg, 0.05 mmol, 0.1 eq.) in nitrogen atmosphere. The vessel was evacuated and refilled with nitrogen (x3), then evacuated, created a hydrogen atmosphere and shaken overnight. The reaction mixture was filtered through Celite (2.5 g), eluted with ethyl acetate, then methanol, and the solvent was removed in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 12 g RediSep™ silica gel cartridge) eluting with a gradient of 0-3% methanol in dichloromethane gave the title product as a yellow gum (286 mg, 0.42 mmol, 84%).

LC/MS (C ₃ 2H46N6O ₅ SiS2) 687 [M+H]+; RT 1.58 (LCMS-V-BI).

¹ H NMR (400 MHz, DMSO^) δ 7.85 (d, 1H), 7.66 (s, 1H), 7.49-7.41 (m, 2H), 7.29-7.22 (m, 1H), 5.86 (s, 2H), 4.39 (t, J=7.2 Hz, 2H), 4.28 (q, J=7.1 Hz, 2H), 3.77-3.68 (m, 2H), 3.48-3.36 (m, 4H), 3.27 (s, 3H), 3.24 (s, 3H), 3.16-3.09 (m, 2H), 2.46 (s, 3H), 2.02-1.93 (m, 2H), 1.92-1.82 (m, 2H), 1.31 (t, J=7.1 Hz, 3h ), 0.97-0.88 (m, 2H), 0.00 (s, 9H).

Step C: ethyl 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(3methoxypropyl)amino)-5-(3-methoxypropyl)-1,3 -thiazole-4-carboxylate.

Hydrochloric acid (4 M in 1,4-dioxane; 5 mL, 20 mmol, 48 eq.) was added to a stirred solution of the product from step B (286 mg, 0.42 mmol, 1 eq.) in 1,4-dioxane (2 mL ), and the mixture was stirred at ambient temperature overnight. The reaction mixture was partitioned between ethyl acetate and saturated aqueous sodium bicarbonate, washed with brine, dried (magnesium sulfate) and concentrated in vacuo. Purification by reverse phase automated flash chromatography (CombiFlash Rf, C18 13 g RediSep column) eluting with a gradient of 5-95% acetonitrile (pH 4; formic acid) in water (pH 4; formic acid) gave the desired product as a yellow foam (162 mg, 0.29 mmol, 70%).

LC/MS (C ₂₆ H ₃₂ N ₆ O ₄ S ₂ ) 557 [M+H] ⁺ ; RT 1.28 (LCMS-V-BI).

1H NMR (400 MHz, DMSOM6) δ 11.06 (br s, 1H), 7.94 (s, 1H), 7.67 (br s + s, 2H), 7.40 (t, J=7.5 Hz, 1H), 7.22 (s, 1H), 4.39 (t, J=7.2 Hz, 2H), 4.29 (q, J=7.1 Hz, 2H), 3.49-3.37 (m, 4H), 3.28 (s, 3H), 3.25 (s, 3H), 3.14 (dd, J=8.7, 6.7 Hz, 2H), 2.47 (s, 3H), 2.03-1.93 (m, 2H), 1.96-1.82 (m, 2H), 1.32 (t, J=7.1 Hz, 3H) .

Step D: 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(3-methoxypropyl)amino)5-(3-methoxypropyl)-1,3 -thiazole-4-carboxylic acid.

To a solution of the product from step C (162 mg, 0.29 mmol, 1 eq.) in 1,4-dioxane (4 ml) was added lithium hydroxide monohydrate (60.9 mg, 1.45 mmol, 5 eq.) and the mixture was refluxed at for 7 hours. The reaction mixture was allowed to cool to ambient temperature and concentrated in vacuo, then dissolved in water, acidified with 2N. aqueous solution of hydrochloric acid and re-evaporated. The residue was triturated with water, filtered and dried in vacuo to give the title product as a beige solid (87.8 mg, 0.17 mmol, 57%) [as hydrochloric acid salt].

HRMS-ERI (m/z) [M+H] ⁺ , calculated for C ₂₄ H ₂₉ N ₆ O ₄ S ₂ : 529.1686; found: 529.1685.

Example 65: 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(3-methoxypropyl)amino)5-(3-phenoxypropyl)-1,3 -thiazole-4-carboxylic acid

Step A: ethyl 2-[(3-methoxypropyl)(5-methyl-6-{[(2Z)-3-{[2-(trimethylsilyl)ethoxy]methyl}-2,3dihydro-1,3-benzothiazol-2 -ylidene] amino } pyridazin-3 -yl) amino] -5-(3 -phenoxyprop-1 -yn-1 -yl)-1,3thiazol-4-carboxylate.

Tetrakis(triphenylphosphine)palladium(0) (83.3 mg, 0.08 mmol, 0.1 equiv.) was added under a nitrogen atmosphere to a stirred solution of synthesis product 11c (500 mg, 0.72 mmol, 1 equiv.), phenylpropargyl ether (0.36 ml, 2.88 mmol, 4 equiv.), triethylamine (0.6 ml, 4.32 mmol, 6 equiv.) and copper iodide (27.5 mg, 0.14 mmol, 0.2 equiv.) in dimethylformamide (10 ml), and the mixture was heated at 100°C overnight. The reaction mixture was allowed to cool to ambient temperature and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 24 g RediSep™ silica gel cartridge) eluting with a gradient of 0-100% ethyl acetate in isoheptane gave the title product as a brown foam (356 mg, 0.48 mmol, 66%).

LC/MS (C ₃₇ H ₄₄ N ₆ O ₅ SiS ₂ ) 745 [M+H] ⁺ ; RT 1.67 (LCMS-V-BI).

- 140 046336

1H NMR (400 MHz, DMSO-de) δ 7.87 (dd, J=7.9, 1.1 Hz, 1H), 7.73 (s, 1H), 7.52-7.42 (m, 2H), 7.38-7.32 (m, 2H), 7.30-7.24 (m, 1H), 7.10-7.05 (m, 2H), 7.00 (tt, J=7.4, 1.1 Hz, 1H), 5.88 (s, 2H), 5.15 (s, 2H), 4.45 (t, J=7.1 Hz, 2H), 4.25 (q, J=7.1 Hz, 2H), 3.77-3.67 (m, 2H), 3.41 (t, J=5.9 Hz, 2H), 3.21 (s, 2H), 2.47 ( s, 3H), 1.98 (p, 2H), 1.28 (t, J=7.1 Hz, 3H), 0.92 (dd, J=8.5, 7.5 Hz, 2H), 0.00 (s, 9H).

Step B: ethyl 2-[(3-methoxypropyl)(5-methyl-6-{[(2Z)-3-{[2-(trimethylsilyl)ethoxy]methyl}-2,3dihydro-1,3-benzothiazol-2 -ylidene]amino}pyridazin-3-yl)amino]-5-(3-phenoxypropyl)-1,3-thiazol-4carboxylate.

A suspension of the product from step A (356 mg, 0.48 mmol, 1 eq.) in ethyl acetate (5 ml) was loaded into a flask containing platinum oxide (PO) (10.8 mg, 0.05 mmol, 0.1 eq.) under a nitrogen atmosphere. The vessel was evacuated and refilled with nitrogen (x3), then evacuated, created a hydrogen atmosphere and shaken overnight. The reaction mixture was filtered through Celite (2.5 g), eluted with ethyl acetate and methanol, the solvent was removed in vacuo and the crude product was purified using automated flash column chromatography (CombiFlash Rf, 12 g RediSep™ silica gel cartridge) eluting with a 0-40% ethyl acetate gradient in isoheptane to obtain the target product in the form of a yellow resin (286 mg, 0.38 mmol, 80%).

LC/MS (C ₃ 7H48N6O5SiS2) 749 [M+H]+; RT 1.66 (ZHMS-U-V1).

1H NMR (400 MHz, DMSO^) δ 7.84 (dd, J=7.7, 1.0 Hz, 1H), 7.66 (s, 1H), 7.50-7.40 (m, 2H), 7.32-7.23 (m, 3H), 6.98 -6.88 (m, 3H), 5.88 (s, 2H), 4.40 (t, J=7.3 Hz, 2H), 4.26 (q, 2H), 4.05 (t, J=6.1 Hz, 2H), 3.78-3.68 ( m, 2H), 3.42 (t, J=6.0 Hz, 2H), 3.31-3.26 (m, 2H), 3.23 (s, 3H), 2.46 (s, 3H), 2.17-2.06 (m, 2H), 2.03 -1.92 (m, 2H), 1.30 (t, J=7.1, 5.2 Hz, 3H), 0.97-0.88 (m, 2H), 0.00 (s, 9H).

Step C: ethyl 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(3methoxypropyl)amino)-5-(3-phenoxypropyl)-1,3 -thiazole-4-carboxylate.

Hydrochloric acid (4 M in 1,4-dioxane; 5 mL, 20 mmol, 52 eq.) was added slowly to a solution of the product from step B (286 mg, 0.38 mmol, 1 eq.) in 1,4-dioxane (2 mL ), and the mixture was stirred at ambient temperature overnight. The reaction mixture was partitioned between ethyl acetate and saturated aqueous sodium bicarbonate, and the organic phase was washed with brine, dried (magnesium sulfate) and concentrated in vacuo. Purification by reverse phase automated flash chromatography (CombiFlash Rf, C18 13 g RediSep column) eluting with a gradient of 5-95% acetonitrile (pH 4; formic acid) in water (pH 4; formic acid) gave the desired product as yellow foam (145 mg, 0.23 mmol, 62%).

LC/MS (C ₃₁ H ₃₄ N ₆ O ₄ S ₂ ) 619 [M+H] ⁺ ; RT 1.40 (LCMS-VB!).

1H NMR (400 MHz, DMSO^) δ 11.07 (br s, 1H), 7.95 (br s, 1H), 7.68 (br s + s, 2H), 7.40 (t, 1H), 7.29 (dd, J=8.8 , 7.3 Hz, 2H), 7.26-7.19 (m, 1H), 7.00-6.89 (m, 3H), 4.39 (t, J=7.2 Hz, 2H), 4.27 (q, J=7.1 Hz, 2H), 4.05 (t, J=6.2 Hz, 2H), 3.44 (t, J=6.0 Hz, 2H), 3.33-3.27 (m, 2H), 3.25 (s, 3H), 2.47 (s, 3H), 2.18-2.06 ( m, 2H), 2.03-1.93 (m, 2H), 1.31 (t, J=7.1 Hz, 3H).

Step D: 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(3-methoxypropyl)amino)5-(3-phenoxypropyl)-1,3 -thiazole-4-carboxylic acid.

To a solution of the product from step C (145 mg, 0.23 mmol, 1 eq.) in 1,4-dioxane (4 ml) was added lithium hydroxide monohydrate (49.3 mg, 1.17 mmol, 5 eq.) and the mixture was heated at 100°C for 7 hours. The reaction mixture was allowed to cool to ambient temperature and concentrated in vacuo, triturated with water, filtered and dried in vacuo to give the title product as a brown solid (90.6 mg, 0.15 mmol, 65%) [as salt lithium].

HRMS-ERI (m/z) [M+H] ⁺ , calculated for C ₂₉ H ₃₁ N ₆ O ₄ S ₂ : 591.1843; found: 591.1843.

Example 66: 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(3-methoxypropyl)amino)5-[3-(cyclohexyloxy)propyl]- 1,3-thiazole-4-carboxylic acid

Step A: ethyl 5-[3-(cyclohexyloxy)prop-1-yn-1-yl]-2-[(3-methoxypropyl)(5-methyl-6-{[(2Z)-3{[2-( trimethylsilyl)ethoxy]methyl } -2,3-dihydro-1,3-benzothiazol-2-ylidene]amino } pyridazin-3 yl)amino]-1,3-thiazol-4-carboxylate.

Tetrakis(triphenylphosphine)palladium(0) (83.3 mg, 0.08 mmol, 0.1 equiv.) under a nitrogen atmosphere was added to a stirred solution of synthesis product 11c (500 mg, 0.72 mmol, 1 equiv.), (prop-2-yn-1yloxy) cyclohexane (400 mg, 2.88 mmol, 4 eq.), triethylamine (0.60 ml, 4.32 mmol, 6 eq.) and iodide

- 141 046336 copper(1) (27.5 mg, 0.14 mmol, 0.2 eq.) in dimethylformamide (10 ml), and the mixture was heated at 100°C overnight. The reaction mixture was allowed to cool to ambient temperature and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 40 g RediSep™ silica gel cartridge) eluting with a gradient of 0-70% ethyl acetate in isoheptane gave the title product as a brown gum (330 mg, 0.44 mmol, 61%).

LC/MS (C ₃₇ H ₅₀ N ₆ O ₅ SiS ₂ ) 751 [M+H]+; RT 1.81 (LCMS-V-BI).

1H NMR (400 MHz, DMSO^) δ 7.86 (d, 1H), 7.73 (s, 1H), 7.51-7.39 (m, 2H), 7.27 (ddd, J=8.3, 7.1, 1.4 Hz, 1H), 5.88 (s, 2H), 4.50 (s, 2H), 4.46 (t, 2H), 4.30 (q, J=7.1 Hz, 2H), 3.73 (t, 2H), 3.60-3.50 (m, 1H), 3.42 ( t, J=6.0 Hz, 2H), 3.23 (s, 3H), 2.48 (s, 3H), 1.98 (p, J=6.3 Hz, 2H), 1.94-1.86 (m, 2H), 1.75-1.63 (m , 2H), 1.541.45 (m, 2H), 1.36-1.17 (m, 7H), 0.92 (dd, J=8.4, 7.6 Hz, 2H), 0.00 (s, 9H).

Step B: ethyl 5-[3-(cyclohexyloxy)propyl]-2-[(3-methoxypropyl)(5-methyl-6-{[(22)-3-{[2(trimethylsilyl)ethoxy]methyl}-2 ,3-dihydro-1,3-benzothiazol-2-ylidene]amino}pyridazin-3-yl)amino]1,3-thiazol-4-carboxylate.

A suspension of the product from step A (330 mg, 0.44 mmol, 1 eq.) in a 1:1 mixture of ethyl acetate/methanol (10 ml) was loaded into a flask containing platinum(^) oxide (9.98 mg, 0.04 mmol, 0.1 eq.) in nitrogen atmosphere. The vessel was evacuated and refilled with nitrogen (x3), then evacuated, created a hydrogen atmosphere and shaken overnight. The reaction mixture was filtered through celite (2.5 g), eluted with ethyl acetate and methanol, and the solvent was removed in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 12 g RediSep™ silica gel cartridge) eluting with a gradient of 0-40% ethyl acetate in isoheptane gave the title product as a yellow gum (272 mg, 0.36 mmol, 82%).

LC/MS (C ₃ 7H54N6O5SiS2) 755 [M+H] ⁺ ; RT 1.79 (LCMS-V-BI).

1H NMR (400 MHz, DMSO^) δ 7.82 (d, J=7.7 Hz, 1H), 7.66 (s, 1H), 7.51-7.40 (m, 2H), 7.26 (ddd, J=8.2, 6.9, 1.6 Hz , 1H), 4.38 (t, 2H), 4.27 (q, J=7.0 Hz, 2H), 3.71 (t, 2H), 3.50-3.38 (m, 4H), 3.24 (s, 3H), 3.23-3.19 ( m, 1H), 3.14 (t, J=7.6 Hz, 2H), 2.45 (s, 3H), 1.97 (p, 2H), 1.91-1.79 (m, 5H), 1.72-1.58 (m, 2H), 1.52 -1.39 (m, 2H), 1.31 (t, J=7.1 Hz, 3H), 1.28-1.14 (m, 4H), 0.92 (t, 2H), 0.00 (s, 9H).

Step C: ethyl 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(3methoxypropyl)amino)-5-[3-(cyclohexyloxy)propyl]- 1,3-thiazole-4-carboxylate.

Hydrochloric acid (4 M in 1,4-dioxane; 4 mL, 16 mmol, 45 eq.) was added to a stirred solution of the product from step B (272 mg, 0.36 mmol, 1 eq.) in 1,4-dioxane (4 mL ), and the mixture was stirred at ambient temperature overnight. The reaction mixture was partitioned between ethyl acetate and saturated aqueous sodium bicarbonate, and the organic phase was washed with brine, dried (magnesium sulfate) and concentrated in vacuo. Purification by reverse phase automated flash chromatography (CombiFlash Rf, C18 13 g RediSep column) eluting with a gradient of 5-95% acetonitrile (pH 4; formic acid) in water (pH 4; formic acid) gave the desired product as a yellow foam (120 mg, 0.19 mmol, 53%).

LC/MS (C ₃₁ H4 ₀ N ₆ O ₄ S ₂ ) 625 [M+H] ⁺ ; RT 1.45 (LCMS-V-BI).

1H NMR (400 MHz, DMSO^) δ 11.09 (br s, 1H), 7.93 (s, 1H), 7.67 (br s + s, 2H), 7.39 (t, 1H), 7.23 (t, 1H), 4.39 (t, J=7.2 Hz, 2H), 4.28 (q, J=7.1 Hz, 2H), 3.45 (dt, J=9.9, 6.1 Hz, 4H), 3.25 (s, 3H), 3.24-3.20 (m, 1H), 3.15 (t, J=7.6 Hz, 2H), 2.47 (s, 3H), 1.97 (p, J=6.3 Hz, 2H), 1.94-1.78 (m, 4H), 1.74-1.62 (m, 2H ), 1.54-1.41 (m, 2H), 1.32 (t, J=7.1 Hz, 3H), 1.29-1.16 (m, 4H).

Step D: 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(3-methoxypropyl)amino)5 - [3-(cyclohexyloxy)propyl] - 1,3-thiazole-4-carboxylic acid.

To a solution of the product from step C (120 mg, 0.19 mmol, 1 eq.) in 1,4-dioxane (4 ml) was added lithium hydroxide monohydrate (40.3 mg, 0.96 mmol, 5 eq.), and the mixture was heated at 100°C during the night. The reaction mixture was allowed to cool to ambient temperature and concentrated in vacuo, triturated with water, filtered and dried in vacuo to give the title product as a yellow solid (98 mg, 0.16 mmol, 86%) [as lithium salt].

HRMS-ERI (m/z) [M+H] ⁺ , calculated for C ₂₉ H ₃₇ N ₆ O ₄ S ₂ : 597.2312; found: 597.2314.

Example 67: 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(3-methoxypropyl)amino)5-[3-(2-fluorophenoxy)propyl ]-1,3-thiazole-4-carboxylic acid

Step A: ethyl 5-(3-hydroxyprop-1-yn-1-yl)-2-[(3-methoxypropyl)(5-methyl-6-{[(22)-3-{[2

- 142 046336 (trimethylsilyl)ethoxy]methyl}-2,3-dihydro-1,3-benzothiazol-2-ylidene]amino}pyridazin-3-yl)amino]1,3-thiazol-4-carboxylate.

Tetrakis(triphenylphosphine)palladium(0) (83.3 mg, 0.08 mmol, 0.1 equiv.) was added under a nitrogen atmosphere to a stirred solution of synthesis product 11c (500 mg, 0.72 mmol, 1 equiv.), propargyl alcohol (0.16 ml, 2.88 mmol, 4 equiv.), triethylamine (0.60 ml, 4.32 mmol, 6 equiv.) and copper(1) iodide (27.5 mg, 0.14 mmol, 0.2 equiv.) in dimethylformamide (10 ml), and the mixture was heated at 100°C for nights. The reaction mixture was allowed to cool to ambient temperature and then concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 40 g RediSep™ silica gel cartridge) eluting with a gradient of 0-100% ethyl acetate in isoheptane provided the title product as a brown solid (362 mg, 0.54 mmol, 75%) .

LC/MS (C ₃₁ H ₄₀ N ₆ O ₅ SiS ₂ ) 669 [M+H]+; RT 1.45 (ZHMS-U-B1).

1H NMR (400 MHz, DMSOD ₆ ) δ 7.88 (dd, J=7.4, 1.1 Hz, 1H), 7.72 (s, 1H), 7.51-7.41 (m, 2H), 7.27 (ddd, J=8.3, 7.1, 1.4 Hz, 1H), 5.89 (s, 2H), 5.44 (t, J=6.0 Hz, 1H), 4.45 (t, J=7.2 Hz, 2H), 4.40 (d, J=5.9 Hz, 2H), 4.29 (q, J=7.1 Hz, 2H), 3.77-3.68 (m, 2H), 3.42 (t, J=6.0 Hz, 2H), 3.24 (s, 3H), 2.48 (s, 3H), 1.99 (p, J=6.3 Hz, 2H), 1.33 (t, J=7.1 Hz, 3H), 0.99-0.88 (m, 2H), 0.00 (s, 9H).

Step B: ethyl 5-(3-hydroxypropyl)-2-[(3-methoxypropyl)(5-methyl-6-{[(22)-3-{[2(trimethylsilyl)ethoxy]methyl}-2,3- dihydro-1,3-benzothiazol-2-ylidene]amino}pyridazin-3-yl)amino]1,3-thiazol-4-carboxylate.

A suspension of the product from step A (362 mg, 0.54 mmol, 1 eq.) in a 1:1 mixture of ethyl acetate/methanol (10 ml) was loaded into a flask containing platinum(IV) oxide (12.3 mg, 0.05 mmol, 0.1 eq.) in nitrogen atmosphere. The vessel was evacuated and refilled with nitrogen (x3), then evacuated, created a hydrogen atmosphere and shaken overnight. The mixture was filtered through celite (2.5 g), eluted with methanol and the solvent was removed in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 24 g RediSep™ silica gel cartridge) eluting with a gradient of 0-80% ethyl acetate in isoheptane gave the title product as a creamy foam (230 mg, 0.34 mmol, 63%).

LC/MS (C ₃₁ H ₄₄ N ₆ O ₅ SiS ₂ ) 673 [M+H]+; RT 1.47 (ZHMS-U-B1).

1H NMR (400 MHz, DMSOL) δ 7.86 (dd, J=7.5, 1.1 Hz, 1H), 7.66 (s, 1H), 7.51-7.40 (m, 2H), 7.25 (ddd, J=8.3, 6.9, 1.6 Hz, 1H), 5.87 (s, 2H), 4.57 (t, J=5.1 Hz, 1H), 4.39 (t, J=7.2 Hz, 2H), 4.28 (q, J=7.1 Hz, 2H), 3.72 ( dd, J=8.5, 7.5 Hz, 2H), 3.52-3.44 (m, 2H), 3.43 (t, 2H), 3.23 (s, 3H), 3.16-3.08 (m, 2H), 2.46 (s, 3H) , 1.96 (p, J=6.4 Hz, 2H), 1.86-1.75 (m, 2H), 1.31 (t, J=7.1 Hz, 3H), 0.92 (dd, J=8.6, 7.4 Hz, 2H), 0.00 ( s, 9H).

Step C: ethyl 5-(3-iodopropyl)-2-[(3-methoxypropyl)(5-methyl-6-{[(22)-3-{[2(trimethylsilyl)ethoxy]methyl}-2,3- dihydro-1,3-benzothiazol-2-ylidene]amino}pyridazin-3-yl)amino]1,3-thiazol-4-carboxylate.

To a stirred solution of the product from step B (230 mg, 0.34 mmol, 1 eq.) in a 3:1 mixture of diethyl ether/acetonitrile (16 ml) was added imidazole (34.9 mg, 0.51 mmol, 1.5 eq.), triphenylphosphine (135 mg, 0.51 mmol, 1.5 eq.) and iodine (130 mg, 0.51 mmol, 1.5 eq.), and the mixture was stirred at ambient temperature overnight. The reaction mixture was diluted with ethyl acetate, washed with 10% aqueous sodium thiosulfate followed by water, then brine, dried (magnesium sulfate) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 12 g RediSep™ silica gel cartridge) eluting with a gradient of 0-50% ethyl acetate in isoheptane gave the title product as a yellow gum (129 mg, 0.16 mmol, 48%).

LC/MS (C ₃₁ H ₄₃ IN ₆ O ₄ SiS ₂ ) 783 [M+H]+; RT 1.63 (ZHMS-U-B1).

1H NMR (400 MHz, DMSO-de) δ 7.85 (dd, J=7.5, 1.1 Hz, 1H), 7.67 (s, 1H), 7.49-7.41 (m, 2H), 7.26 (ddd, J=8.3, 6.9 , 1.6 Hz, 1H), 5.87 (s, 2H), 4.40 (t, J=7.3 Hz, 2H), 4.29 (q, J=7.1 Hz, 2H), 3.73 (dd, J=8.6, 7.4 Hz, 2H ), 3.42 (t, J=6.0 Hz, 2H), 3.40-3.28 (m, 2H), 3.24 (s, 3H), 3.21-3.14 (m, 2H), 2.45 (s, 3H), 2.14 (p, J=7.0 Hz, 2H), 2.02-1.95 (m, 2H), 1.34 (t, J=7.1 Hz, 3H), 0.92 (dd, J=8.6, 7.4 Hz, 2H), 0.00 (s, 9H).

Step D: Ethyl 5-[3-(2-fluorophenoxy)propyl]-2-[(3-methoxypropyl)(5-methyl-6-{[(22)-3-{[2(trimethylsilyl)ethoxy]methyl} -2,3-dihydro-1,3-benzothiazol-2-ylidene]amino}pyridazin-3-yl)amino]1,3-thiazol-4-carboxylate.

Sodium hydride (60% in mineral oil; 13.2 mg, 0.33 mmol, 2 eq.) was added to a solution of 2fluorophenol (0.02 mL, 0.2 mmol, 1.2 eq.) in dimethylformamide (3 mL). After 15 min, the mixture was cooled to 0°C and a solution of the product from step C (129 mg, 0.16 mmol, 1 eq.) in dimethylformamide (3 ml) was added. The mixture was allowed to warm to ambient temperature and stirred overnight. The reaction mixture was quenched by adding water, acidified with 2N. aqueous hydrochloric acid solution and extracted with ethyl acetate (x2). The combined organic extracts were washed with brine, dried (magnesium sulfate) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 12 g RediSep™ silica gel cartridge) eluting with a gradient of 0-6% methanol in dichloromethane gave the title product as a creamy gum (75.6 mg, 0.1 mmol, 60%).

LC/MS (C ₃₇ H ₄₇ FN ₆ O ₅ SiS ₂ ) 767 [M+H]+; RT 1.61 (ZHMS-U-B1).

- 143 046336

1H NMR (400 MHz, DMSO-dg) δ 7.84 (d, 1H), 7.67 (s, 1H), 7.51-7.40 (m, 2H), 7.30-7.07 (m, 4H), 6.976.89 (m, 1H ), 5.87 (s, 2H), 4.40 (t, J=7.2 Hz, 2H), 4.26 (q, J=7.0 Hz, 2H), 4.13 (t, J=6.1 Hz, 2H), 3.78-3.69 (m , 2H), 3.42 (t, J=6.0 Hz, 2H), 3.31-3.25 (m, 2H), 3.24 (s, 3H), 2.46 (s, 3H), 2.25-2.05 (m, 2H), 1.97 ( p, J=6.2 Hz, 2H), 1.29 (t, J=7.1 Hz, 3H), 0.98-0.89 (m, 2H), 0.00 (s, 9H).

Step E: ethyl 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(3methoxypropyl)amino)-5-[3 -(2-fluorophenoxy)propyl ]-1,3-thiazole-4-carboxylate.

Hydrochloric acid (4 M in 1,4-dioxane; 1 mL, 4 mmol, 40.6 eq.) was added to a stirred solution of the product from step D (75.6 mg, 0.1 mmol, 1 eq.) in 1,4-dioxane (2 mL ), and the mixture was stirred at ambient temperature overnight. The reaction mixture was partitioned between ethyl acetate and saturated aqueous sodium bicarbonate, and the organic phase was washed with brine, dried (magnesium sulfate) and concentrated in vacuo. Purification by reverse phase automated flash chromatography (CombiFlash Rf, C18 13 g RediSep column) eluting with a gradient of 5-95% acetonitrile (pH 4; formic acid) in water (pH 4; formic acid) gave the desired product in the form of a yellow glassy substance (36.2 mg, 0.06 mmol, 58%).

LC/MS (C ₃₁ H ₃₃ FN ₆ O ₄ S ₂ ) 637 [M+H]+; RT 1.39 (LCMS-V-BI).

1H NMR (400 MHz, DMSO-dg) 7.92 (br s, 1H), 7.67 (br s + s, 2H), 7.39 (t, J=7.5 Hz, 1H), 7.27-7.08 (m, 4H), 6.99 -6.89 (m, 1H), 4.39 (t, J=7.3 Hz, 2H), 4.26 (q, J=7.1 Hz, 2H), 4.14 (t, J=6.1 Hz, 2H), 3.44 (t, J= 6.0 Hz, 2H), 3.34-3.24 (m, 2H), 3.25 (s, 3H), 2.47 (s, 2H), 2.22-2.06 (m, 2H), 1.97 (p, 2H), 1.30 (t, J =7.1 Hz, 3H).

Step F: 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3 -yl} (3-methoxypropyl)amino)-5[3-(2-fluorophenoxy)propyl ] -1,3-thiazole-4-carboxylic acid.

To a solution of the product from step E (36.2 mg, 0.06 mmol, 1 eq.) in 1,4-dioxane (3 ml) was added lithium hydroxide monohydrate (11.9 mg, 0.28 mmol, 5 eq.) and the mixture was refluxed at during the night. The reaction mixture was allowed to cool to ambient temperature and concentrated in vacuo. Purification by reverse phase automated flash chromatography (CombiFlash Rf, C18 13 g RediSep column) eluting with a gradient of 5-95% acetonitrile (pH 4; formic acid) in water (pH 4; formic acid) gave the desired product as a beige solid (19.5 mg, 0.03 mmol, 56%).

MSHR-ERI (m/z) [M+H]+, calculated for C ₂₉ H ₃₀ FN ₆ O ₄ S ₂ : 609.1749; found: 609.1753.

Example 68: 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl} (methyl)amino)-5-[3 (benzyloxy)azetidin-1 -yl ]-1,3-thiazole-4-carboxylic acid

Step A: ethyl 5-[3-(benzyloxy)azetidin-1-yl]-2-[methyl(5-methyl-6-{[(27)-3-{[2(trimethylsilyl)ethoxy]methyl}-2 ,3-dihydro-1,3-benzothiazol-2-ylidene]amino}pyridazin-3-yl)amino]1,3-thiazol-4-carboxylate.

Synthesis product 11b (500 mg, 0.79 mmol, 1 eq.), 3-(benzyloxy)azetidine hydrochloride (314 mg, 1.57 mmol, 2 eq.), cesium carbonate (769 mg, 2.36 mmol, 3 equiv.) and 1,4-dioxane (10 ml). The mixture was bubbled with nitrogen (10 min), after which xanthos Pd G3 (74.7 mg, 0.08 mmol, 0.1 eq.) was added and the mixture was heated at 140°C for 2 h under microwave irradiation.

The reaction mixture was diluted with ethyl acetate, then washed with water followed by brine, and the organic phase was dried (magnesium sulfate) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 24 g RediSep™ silica gel cartridge) eluting with a gradient of 0-50% ethyl acetate in isoheptane gave the title product as a yellow foam (380 mg, 0.53 mmol, 67%).

LC/MS (C ₃₅ H ₄₃ N ₇ O ₄ SiS ₂ ) 718 [M+H]+; RT 1.53 (LCMS-V-BI).

1H NMR (400 MHz, DMSO-dg) δ 7.82 (d, 1H), 7.57 (d, J=1.0 Hz, 1H), 7.50-7.42 (m, 2H), 7.40-7.35 (m, 4H), 7.34- 7.28 (m, 1H), 7.27-7.21 (m, 1H), 5.84 (s, 2H), 4.52 (s, 2H), 4.52-4.44 (m, 1H), 4.32 (ddd, J=8.9, 6.3, 1.2 Hz, 2H), 4.19 (q, J=7.1 Hz, 2H), 4.03-3.94 (m, 2H), 3.76-3.70 (m, 2H), 3.66 (s, 3H), 2.44 (d, J=1.0 Hz , 3H), 1.26 (t, J=7.1 Hz, 3H), 0.97-0.88 (m, 2H), 0.00 (s, 9H).

Step B: ethyl 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)-5-[3(benzyloxy)azetidin-1- yl]-1,3-thiazol-4-carboxylate.

Trifluoroacetic acid (1 ml, 13.4 mmol, 96.3 eq.) was added to the stirred solution

- 144 046336 product from step A (100 mg, 0.14 mmol, 1 eq.) in dichloromethane (3 ml), and the mixture was stirred at ambient temperature overnight. The reaction mixture was diluted with dichloromethane and washed with saturated aqueous sodium bicarbonate, then brine, dried (magnesium sulfate) and concentrated in vacuo. Purification by reverse phase automated flash chromatography (CombiFlash Rf, C18 13 g RediSep column) eluting with a gradient of 5-95% acetonitrile (pH 4; formic acid) in water (pH 4; formic acid) gave the desired product as yellow glassy substance (48 mg, 0.08 mmol, 59%).

LC/MS (C ₂₉ H ₂₉ N ₇ O ₃ S ₂ ) 588 [M+H]+; RT 1.31 (LCMS-V-BI).

¹ H NMR (400 MHz, DMSOHO δ 11.02 (br s, 1H), 7.90 (br s, 1H), 7.59 (br s + s, 2H), 7.44-7.28 (m, 6H), 7.21 (t, J= 7.5 Hz, 1H), 4.52 (s, 2H), 4.51-4.46 (m, 1H), 4.37-4.28 (m, 2H), 4.20 (q, J=7.1 Hz, 2H), 4.01-3.96 (m, 2H ), 3.66 (s, 3H), 2.45 (s, 3H), 1.27 (t, J=7.1 Hz, 3H).

Step C: 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl} (methyl)amino)-5-[3 (benzyloxy)azetidin-1 -yl ] -1,3-thiazole-4-carboxylic acid.

To a solution of the product from step B (48 mg, 0.08 mmol, 1 eq.) in 1,4-dioxane (4 ml) was added lithium hydroxide monohydrate (17.1 mg, 0.41 mmol, 5 eq.), and the mixture was heated at 100°C during the night. The reaction mixture was allowed to cool to ambient temperature and concentrated in vacuo. The residue was triturated with water, filtered and dried in vacuo to give the title product as a beige solid (33.2 mg, 0.06 mmol, 73%) [as lithium salt].

HRMS-ERI (m/z) [M+H] ⁺ , calculated for C ₂₇ H ₂₆ N ₇ O ₃ S ₂ : 560.1533; found: 560.1531.

Example 69: 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3 -yl} (methyl)amino)-5 -(3 phenoxyazetidin-1 -yl)-1 ,3-thiazole-4-carboxylic acid kg s

Step A: ethyl 2-[methyl(5-methyl-6-{[(2Z)-3-{[2-(trimethylsilyl)ethoxy]methyl}-2,3-dihydro-1,3benzothiazol-2-ylidene]amino }pyridazin-3 -yl)amino]-5-(3-phenoxyazetidin-1 -yl)-1,3-thiazol-4 carboxylate.

Synthesis product 11b (100 mg, 0.16 mmol, 1 eq.), 3-phenoxyazetidine hydrochloride (58.4 mg, 0.31 mmol, 2 eq.), cesium carbonate (154 mg, 0.47 mmol, 3 eq.) were loaded into a vessel dried in an oven. ) and 1,4-dioxane (4 ml). The mixture was bubbled with nitrogen (10 min) and rac-BINAP Pd G3 (15.6 mg, 0.02 mmol, 0.1 eq.) was added and the mixture was heated at 100°C for 24 h. The reaction mixture was diluted with ethyl acetate, washed with water, followed by brine. , dried (magnesium sulfate) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 12 g RediSep™ silica gel cartridge) eluting with a gradient of 0-50% ethyl acetate in isoheptane gave the title product as a yellow glass (55.7 mg, 0.08 mmol, 50%).

LC/MS (C ₃₄ H ₄₁ N ₇ O ₄ SiS ₂ ) 704 [M+H] ⁺ ; RT 1.60 (LCMS-V-BI).

1H NMR (400 MHz, DMSO) δ 7.82 (d, 1H), 7.59 (d, J=1.1 Hz, 1H), 7.51-7.39 (m, 2H), 7.39-7.30 (m, 2H), 7.29-7.20 ( m, 1H), 7.00 (tt, J=7.4, 1.1 Hz, 1H), 6.91 (dt, J=7.7, 1.1 Hz, 2H), 5.85 (s, 2H), 5.16 (tt, J=6.6, 3.8 Hz , 1H), 4.62-4.53 (m, 2H), 4.22 (q, J=7.1 Hz, 2H), 4.18-4.09 (m, 2H), 3.73 (dd, J=8.5, 7.5 Hz, 2H), 3.69 ( s, 3H), 2.45 (d, J=1.0 Hz, 3H), 1.28 (t, J=7.1 Hz, 3H), 0.97-0.88 (m, 2H), 0.00 (s, 9H).

Step B: ethyl 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)-5-(3phenoxyazetidin-1-yl)-1 ,3-thiazole-4-carboxylate.

Trifluoroacetic acid (1 mL, 13.4 mmol, 170 eq.) was added to a stirred solution of the product from Step A (55.7 mg, 0.08 mmol, 1 eq.) in dichloromethane (3 mL), and the mixture was stirred at ambient temperature overnight. The reaction mixture was diluted with dichloromethane, washed with saturated aqueous sodium bicarbonate, then with brine, dried (magnesium sulfate) and concentrated in vacuo. Purification by reverse phase automated flash chromatography (CombiFlash Rf, C18 13 g RediSep column) eluting with a gradient of 5-95% acetonitrile (pH4; formic acid) in water (pH 4; formic acid) gave the desired product in in the form of a yellow glassy substance (20.4 mg, 0.04 mmol, 45%).

LC/MS (C ₂₈ H ₂₇ N ₇ O ₃ S ₂ ) 574 [M+H] ⁺ ; RT 1.31 (LCMS-V-BI).

1H NMR (400 MHz, DMSO-ds) δ 7.92 (br s, 1H), 7.60 (br s + s, 2H), 7.41-7.30 (m, 3H), 7.21 (s, 1H), 7.01 (tt, J =7.4, 1.1 Hz, 1H), 6.91 (dt, J=7.8, 1.1 Hz, 2H), 5.16 (tt, J=6.5, 3.7 Hz, 1H), 4.62-4.53 (m, 2H), 4.22 (q, J=7.1 Hz, 2H), 4.17-4.10 (m, 2H), 3.68 (s, 3H), 2.45 (s, 2H), 1.28 (t, J=7.1 Hz, 3H).

Step C: 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3 -yl} (methyl)amino)-5 -(3 phenoxyazetidin-1 -yl)-1 ,3-thiazole-4-carboxylic acid.

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To a solution of the product from step B (20 mg, 0.03 mmol, 1 eq.) in 1,4-dioxane (3 ml) was added lithium hydroxide monohydrate (7.31 mg, 0.17 mmol, 5 eq.), and the mixture was heated at 100°C during the night. The reaction mixture was allowed to cool to ambient temperature, concentrated in vacuo, triturated with water, filtered and dried in vacuo to give the title product as a beige solid (11.2 mg, 0.02 mmol, 59%) [as lithium salt].

HRMS-ERI (m/z) [M+H]+, calculated for C ₂₆ H ₂₄ N ₇ O ₃ S ₂ : 546.1377; found: 546.1381.

Example 70: 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)-5-[(3S)-3(benzyloxy)pyrrolidine -1-yl]-1,3-thiazol-4-carboxylic acid

Step A: ethyl 5-[(3S)-3-(benzyloxy)pyrrolidin-1-yl]-2-[methyl(5-methyl-6-{[(2Z)-3-{[2(trimethylsilyl)ethoxy] methyl}-2,3-dihydro-1,3-benzothiazol-2-ylidene]amino}pyridazin-3-yl)amino]1,3-thiazol-4-carboxylate.

Synthesis product 11b (200 mg, 0.31 mmol, 1 eq.), N-3-benzyloxypyrrolidine hydrochloride (135 mg, 0.63 mmol, 2 eq.), cesium carbonate (308 mg, 0.94 mmol, 3 equiv.) and 1,4-dioxane (8 ml). The mixture was bubbled with nitrogen (10 min), after which rac-BINAP Pd G3 (15.6 mg, 0.02 mmol, 0.05 eq.) was added and the mixture was heated at 100°C for 24 h. The reaction mixture was diluted with ethyl acetate, then washed with water, followed by washed with brine, dried (magnesium sulfate) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 12 g RediSep™ silica gel cartridge) eluting with a gradient of 0-50% ethyl acetate in isoheptane gave the title product as a yellow foam (113 mg, 0.15 mmol, 49%).

LC/MS (C ₃₆ H ₄₅ N ₇ O ₄ SiS ₂ ) 732 [M+H]+; RT 1.54 (LCMS-V-BI).

¹ H NMR (400 MHz, DMSO-dg) δ 7.82 (d, 1H), 7.57 (d, 1H), 7.49-7.40 (m, 2H), 7.36-7.31 (m, 4H), 7.317.22 (m, 2H), 5.84 (s, 2H), 4.60-4.48 (m, 2H), 4.30-4.26 (m, 1H), 4.25-4.17 (m, 2H), 3.71 (t, 2H), 3.68 (s, 3H) , 3.64-3.52 (m, 2H), 3.48-3.36 (m, 2H), 2.44 (d, J=1.0 Hz, 3H), 2.14 (s, 2H), 1.29 (t, J=7.1 Hz, 3H), 0.97-0.88 (m, 2H), 0.00 (s, 9H).

Step B: ethyl 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)-5-[(38)3-(benzyloxy) pyrrolidin-1-yl]-1,3-thiazol-4-carboxylate.

Hydrochloric acid (4 M in 1,4-dioxane; 4 mL, 16 mmol, 104 eq.) was added to a stirred solution of the product from step A (113 mg, 0.15 mmol, 1 eq.) in 1,4-dioxane (5 mL ), and the mixture was stirred at 40°C overnight. The reaction mixture was diluted with ethyl acetate, washed with saturated aqueous sodium bicarbonate, then with brine, dried (magnesium sulfate) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 12 g RediSep™ silica gel cartridge) eluting with a gradient of 0-5% methanol in dichloromethane gave the title product as a yellow glass (83 mg, 0.14 mmol, 89%) .

LC/MS (C ₃₀ H ₃₁ N ₇ O ₃ S ₂ ) 602 [M+H]+; RT 1.32 (LCMS-V-BI).

1H NMR (400 MHz, DMSO-dg) δ 10.96 (br s, 1H), 7.96 (br s, 1H), 7.60 (br s + s, 2H), 7.42-7.23 (m, 5H), 7.21 (s, 2H), 4.61-4.48 (m, 2H), 4.33-4.26 (m, 1H), 4.25-4.16 (m, 2H), 3.73 (dd, J=11.4, 4.6 Hz, 1H), 3.67 (s, 3H) , 3.64-3.53 (m, 1H), 3.45-3.36 (m, 2H), 2.45 (s, 3H), 2.26-2.05 (m, 2H), 1.29 (t, J=7.1 Hz, 3H).

Step C: 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)-5-[(3S)-3(benzyloxy)pyrrolidine -1-yl]-1,3-thiazol-4-carboxylic acid.

To a solution of the product from step B (83 mg, 0.14 mmol, 1 eq.) in 1,4-dioxane (8 ml) was added lithium hydroxide monohydrate (28.9 mg, 0.69 mmol, 5 eq.), and the mixture was heated at 100°C during the night. The reaction mixture was allowed to cool to ambient temperature and concentrated in vacuo, triturated with water, filtered and dried in vacuo to give the title product as a yellow solid (65.6 mg, 0.11 mmol, 83%).

MSHR-ERI (m/z) [M+H] ⁺ , calculated for C28H28N7O3S2: 574.1690; found: 574.1687.

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Example 71: 2-({6-[(1,3-benzothiαzol-2-yl)amino]-5-methylpyridαzin-3-yl}(methyl)amino)-5-[(3R)-3(benzyloxy)pyrrolidine -1-yl]-1,3-thiazol-4-carboxylic acid

Step A: ethyl 5-[(3R)-3-(benzyloxy)pyrrolidin-1-yl]-2-[methyl(5-methyl-6-{[(2Z)-3-{[2(trimethylsilyl)ethoxy] methyl } -2,3-dihydro-1,3-benzothiazol-2-ylidene] amino } pyridazin-3 -yl) amino] 1,3-thiazol-4-carboxylate.

Synthesis product 11b (200 mg, 0.31 mmol, 1 eq.), N-3-benzyloxypyrrolidine hydrochloride (135 mg, 0.63 mmol, 2 eq.), cesium carbonate (308 mg, 0.94 mmol, 3 equiv.) and 1,4-dioxane (8 ml). The mixture was bubbled with nitrogen (10 min), after which rac-BINAP Pd G3 (31.2 mg, 0.03 mmol, 0.1 eq.) was added and the mixture was heated at 100°C for 24 h. The reaction mixture was diluted with ethyl acetate, washed with water, followed by washing brine, dried (magnesium sulfate) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 12 g RediSep™ silica gel cartridge) eluting with a gradient of 0-50% ethyl acetate in isoheptane gave the title product as a yellow glass (101 mg, 0.14 mmol, 44%) .

LC/MS (C36H4 ₅ N ₇ O4SiS2) 732 [M+H]+; RT 1.55 (LCMS-V-BI).

1H NMR (400 MHz, DMSO-de) δ 7.82 (d, 1H), 7.57 (d, J=1.1 Hz, 1H), 7.49-7.38 (m, 2H), 7.36-7.31 (m, 4H), 7.30- 7.21 (m, 2H), 5.84 (s, 2H), 4.60-4.47 (m, 2H), 4.32-4.25 (m, 1H), 4.25-4.16 (m, 2h), 3.77-3.68 (m, 2h), 3.67 (s, 3H), 3.62-3.52 (m, 2H), 3.45-3.36 (m, 2H), 2.43 (s, 3H), 2.22-2.09 (m, 2H), 1.29 (t, J=7.1 Hz, 3H), 0.91 (dd, J=8.4, 7.5 Hz, 2H), 0.00 (s, 9H).

Step B: ethyl 2-({6-[(1,3-benzothiαzol-2-yl)amino]-5-methylpyridαzin-3-yl}(methyl)amino)-5-[(3R)3-(benzyloxy) pyrrolidin-1-yl]-1,3-thiazol-4-carboxylate.

Hydrochloric acid (4 M in 1,4-dioxane; 4 mL, 16 mmol, 117 eq.) was added to a stirred solution of the product from step A (101 mg, 0.14 mmol, 1 eq.) in 1,4-dioxane (5 mL ), and the mixture was stirred at 40°C overnight. The reaction mixture was diluted with dichloromethane, washed with saturated aqueous sodium bicarbonate, then with brine, dried (magnesium sulfate) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 12 g RediSep™ silica gel cartridge) eluting with a gradient of 0-5% methanol in dichloromethane gave the title product as a yellow gum (80 mg, 0.13 mmol, 97%).

LC/MS (C ₃₀ H ₃₁ N ₇ O ₃ S ₂ ) 602 [M+H] ⁺ ; RT 1.32 (LCMS-V-BI).

1H NMR (400 MHz, DMSO-de) δ 10.98 (br s, 1H), 7.91 (s, 1H), 7.60 (br s + s, 2H), 7.42-7.11 (m, 7H), 4.60-4.48 (m , 2H), 4.33-4.27 (m, 1H), 4.27-4.13 (m, 2H), 3.67 (s, 3H), 3.63-3.55 (m, 2H), 3.42 (td, J=9.3, 8.7, 3.4 Hz , 2H), 2.46 (s, 3H), 2.25-2.06 (m, 2H), 1.30 (t, J=7.1 Hz, 3H).

Step C: 2-({6-[(1,3-benzothiαzol-2-yl)amino]-5-methylpyridαzin-3-yl}(methyl)amino)-5-[(3R)-3(benzyloxy)pyrrolidine -1-yl]-1,3-thiazol-4-carboxylic acid.

To a solution of the product from step B (80 mg, 0.13 mmol, 1 eq.) in 1,4-dioxane (5 ml) was added lithium hydroxide monohydrate (28 mg, 0.67 mmol, 5 eq.) and the mixture was heated to 100°C during the night. The reaction mixture was allowed to cool to ambient temperature and concentrated in vacuo, triturated with water, filtered and dried in vacuo to give the title product as a yellow solid (50.1 mg, 0.09 mmol, 65%) [as lithium salt].

HRMS-ERI (m/z) [M+H] ⁺ , calculated for C2sH2sN ₇ O3S2: 574.1690; found: 574.1723.

Example 72: 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)-5-(3hydroxyazetidin-1-yl)-1, 3-thiazole-4-carboxylic acid

Step A: ethyl 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)-5-(3hydroxyazetidin-1-yl)-1 ,3-thiazole-4-carboxylate.

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A solution of the product from Example 68, Step B (94.2 mg, 0.16 mmol, 1 eq.) in dichloromethane (5 ml) was cooled to -78°C and boron trichloride (1 M in dichloromethane; 0.48 ml, 0.48 mmol, 3 eq.). The mixture was kept at -78°C for 1 hour and then allowed to warm to ambient temperature over 2 hours. The reaction mixture was quenched by adding saturated aqueous sodium bicarbonate, then extracted with dichloromethane (x2) followed by extraction with a 3:1 mixture dichloromethane/isopropanol, and the combined organic extracts were dried (magnesium sulfate) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 12 g RediSep™ silica gel cartridge) eluting with a gradient of 0-6% methanol in dichloromethane provided the title product as a yellow solid (13.4 mg, 0.03 mmol, 17%) .

LC/MS (C ₂₂ H ₂₃ N ₇ O ₃ S ₂ ) 498 [M+H]+; RT 1.00 (LCMS-V-BI).

¹ H NMR (400 MHz, DMSO) δ 7.87 (d, J=7.7 Hz, 1H), 7.56 (s, 1H), 7.51 (br s, 1H), 7.41-7.32 (m, 1H), 7.18 (t, J=7.6 Hz, 1H), 5.75 (d, 1H), 4.55 (h, J=6.0 Hz, 1H), 4.40-4.31 (m, 2H), 4.21 (q, J=7.1 Hz, 2H), 3.88- 3.80 (m, 2H), 3.65 (s, 3H), 2.43 (s, 3H), 1.28 (t, J=7.1 Hz, 3H).

Step B: 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)-5-(3hydroxyazetidin-1-yl)-1, 3-thiazole-4-carboxylic acid.

To a solution of the product from step C (13.4 mg, 0.03 mmol, 1 eq.) in 1,4-dioxane (5 ml) was added lithium hydroxide monohydrate (5.65 mg, 0.13 mmol, 5 eq.), and the mixture was heated at 100°C during the night. The reaction mixture was allowed to cool to ambient temperature and concentrated in vacuo. Purification by reverse phase automated flash chromatography (CombiFlash Rf, C18 13 g RediSep column) eluting with a gradient of 5-95% acetonitrile in water gave the title product as a beige solid (8.1 mg, 0.02 mmol, 64% ).

HRMS-ERI (m/z) [M+H]+, calculated for C ₂₀ H ₂₀ N ₇ O ₃ S ₂ : 470.1064; found: 470.1072.

Example 73: 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl} (methyl)amino)-5-(3 -{2fluoro-4-[3 -(methylamino)prop-1 -yn-1 -yl]phenoxy}propyl)-1,3-thiazol-4-carboxylic acid

Step A: ethyl 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)-5-(3-{2fluoro-4-[ 3-(methylamino)prop-1-yn-1-yl]phenoxy}propyl)-1,3-thiazol-4-carboxylate.

Trifluoroacetic acid (20 ml) was added to a stirred solution of synthesis 5j, step A (1.5 g, 1.71 mmol, 1 eq.) in dichloromethane (60 ml), and the mixture was stirred at ambient temperature overnight. The reaction mixture was diluted with dichloromethane, cooled to 0°C, then made alkaline by adding 2N. aqueous sodium hydroxide solution, and the organic phase was dried (magnesium sulfate) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 40 g RediSep™ silica gel cartridge) eluting with a gradient of 0-10% methanol in dichloromethane gave the title product as a yellow solid (361 mg, 0.56 mmol, 33%) .

LC/MS (C ₃₂ H ₃₂ FN ₇ O ₃ S ₂ ) 646 [M+H]+; RT 1.98 (LCMS-VC).

1H NMR (400 MHz, DMSO-de) δ 7.91 (d, 1H), 7.68 (d, J=1.2 Hz, 1H), 7.53 (d, J=7.9 Hz, 1H), 7.39 (ddd, J=8.2, 7.2, 1.3 Hz, 1H), 7.32-7.11 (m, 4H), 4.25 (q, J=7.1 Hz, 2H), 4.15 (t, J=6.2 Hz, 2H), 3.77 (s, 3H), 3.46 ( s, 2H), 3.27 (t, J=7.7 Hz, 2H), 2.47 (d, J=1.0 Hz, 3H), 2.31 (s, 3H), 2.19-2.07 (m, 2H), 2.23 (s, 1H ), 1.30 (t, J=7.1 Hz, 3H).

Step B: 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl} (methyl)amino)-5-(3 -{2fluoro-4-[3 -(methylamino)prop-1-yn-1-yl]phenoxy}propyl)-1,3-thiazol-4-carboxylic acid.

To a solution of the product from step B (361 mg, 0.56 mmol, 1 eq.) in 1,4-dioxane (15 ml) was added lithium hydroxide monohydrate (352 mg, 8.39 mmol, 15 eq.) and the mixture was heated at 100°C during the night. The reaction mixture was allowed to cool to ambient temperature and concentrated in vacuo. The residue was triturated with water, filtered, washed with water, then diethyl ether, and dried in vacuo to give the title product as a yellow solid (286 mg, 0.46 mmol, 83%) [as lithium salt].

HRMS-ERI (m/z) [M+H]+, calculated for C ₃₀ H ₂₉ FN ₇ O ₃ S ₂ : 618.1752; found: 618.1767.

Example 74: 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)-5-{3-[2fluoro-4-(3 -{methyl[2-(methylamino)ethyl]amino}prop-1-yn-1-yl)phenoxy]propyl}-1,3-thiazol-4-carboxylic

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Step A: ethyl 5-[3-(4-{3-[(2-{[(tert-butoxy)carbonyl](methyl)amino}ethyl)(methyl)amino]prop-1in-1-yl}-2 -fluorophenoxy)propyl]-2-[methyl(5-methyl-6-{[(^)-3-{[2-(trimethylsilyl)ethoxy]methyl}-2,3dihydro-1,3-benzothiazol-2-ylidene ]amino} pyridazin-3-yl)amino]-1,3-thiazol-4-carboxylate.

NBoc-(methylamino)acetaldehyde (234 mg, 1.35 mmol, 2 equiv.), sodium triacetoxyborohydride (343 mg, 1.62 mmol, 2.4 eq.) and glacial acetic acid (50 μl), and the mixture was stirred at ambient temperature for 6 hours. The reaction mixture was partitioned between ethyl acetate and saturated aqueous sodium bicarbonate, and the organic phase was dried (magnesium sulfate) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 40 g RediSep™ silica gel cartridge) eluting with a gradient of 0-50% ethyl acetate in isoheptane gave the title product as a yellow gum (705 mg, 0.76 mmol, >100%) .

LC/MS (C46H61N8O6FSiS2) 933 [M+H]+; RT 1.39 (ZHMS-U-V2).

¹ H NMR (400 MHz, DMSO-d^ δ 7.84 (d, 1H), 7.67 (d, 1H), 7.51-7.40 (m, 2H), 7.33-7.12 (m, 4H), 5.87 (s, 2H) , 4.25 (q, J=7.1 Hz, 2H), 4.15 (t, J=6.1 Hz, 2H), 3.77 (s, 3H), 3.76-3.68 (m, 2H), 3.49 (s, 2H), 3.31- 3.20 (m, 6H), 2.76 (s, 3H), 2.47 (d, J=1.0 Hz, 3H), 2.26 (s, 3H), 2.18-2.07 (m, 2H), 1.37 (s, 9H), 1.28 (t, J=7.1 Hz, 3H), 0.98-0.87 (m, 2H), 0.00 (s, 9H).

Step B: ethyl 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)-5-{3-[2fluoro-4-( 3-{methyl[2-(methylamino)ethyl]amino}prop-1-yn-1-yl)phenoxy]propyl}-1,3-thiazol-4carboxylate.

Trifluoroacetic acid (5 ml) was added to a stirred solution of the product from step A (705 mg, 0.76 mmol, 1 eq.) in dichloromethane (20 ml), and the mixture was stirred at ambient temperature for 12 hours. The reaction mixture was diluted with dichloromethane, cooled to 0°C and made alkaline by adding 2N. aqueous sodium hydroxide solution, and the organic phase was dried (magnesium sulfate) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 12 g RediSep™ silica gel cartridge) eluting with a gradient of dichloromethane to 20% methanol in dichloromethane gave the title product as a yellow gum (123 mg, 0.17 mmol, 22%).

LC/MS (C ₃₅ H3 ₉ FN ₈ O ₃ S ₂ ) 703 [M+H] ⁺ ; RT 2.07 (LCMS-VC).

1H NMR (400 MHz, DMSO) δ 7.90 (d, 1H), 7.67 (d, J=1.2 Hz, 1H), 7.53 (d, 1H), 7.42-7.36 (m, 1H), 7.34-7.26 (dd, 1H), 7.25-7.12 (m, 3H), 4.27 (q, J=7.1 Hz, 2H), 4.15 (t, J=6.1 Hz, 2H), 3.76 (s, 3H), 3.48 (s, 2h), 3.30-3.20 (m, 2H), 2.60 (dd, J=6.7, 5.2 Hz, 2H), 2.51-2.46 (m, 2H), 2.46 (s, 3H), 2.30 (s, 3H), 2.23 (s, 3H), 2.192.08 (m, 2H), 1.30 (t, J=7.1 Hz, 3H).

Step C: 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)-5-{3-[2fluoro-4-(3 -{methyl[2-(methylamino)ethyl]amino}prop-1-yn-1-yl)phenoxy]propyl}-1,3-thiazol-4-carboxylic acid.

To a solution of the product from step B (123 mg, 0.17 mmol, 1 eq.) in 1,4-dioxane (15 ml) was added lithium hydroxide monohydrate (110 mg, 2.62 mmol, 15 eq.) and the mixture was heated to 100°C during the night. The reaction mixture was allowed to cool to ambient temperature and concentrated in vacuo. Purification by reverse phase automated flash chromatography (CombiFlash Rf, C18 13 g RediSep column) eluting with a gradient of 5-95% acetonitrile (pH 9; ammonium formate) in water (pH 9; ammonium formate) provided the desired product as a yellow solid (75.8 mg, 0.11 mmol, 64%) [as lithium salt].

HRMS-ERI (m/z) [M+H] ⁺ , calculated for C ₃₃ H ₃₆ FN ₈ O ₃ S ₂ : 675.2330; found: 675.2331.

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Example 75: 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)-5-[4(benzyloxy)piperidin-1 -yl ]-1,3-thiazole-4-carboxylic acid

Step A: Ethyl 5-[4-(benzyloxy)piperidin-1-yl]-2-[(6-chloro-5-methylpyridazin-3yl)(methyl)amino]-1,3-thiazol-4-carboxylate.

Sodium hydride (60% in mineral oil; 27.2 mg, 0.68 mmol, 1.2 eq.) was added to a cooled solution of the synthesis product 3za (213 mg, 0.57 mmol, 1 eq.) and 3,6-dichloro-4-methylpyridazine (92.5 mg , 0.57 mmol, 1 eq.) in tetrahydrofuran (6 ml), and the mixture was allowed to warm to ambient temperature and stirred overnight. The reaction mixture was diluted with dichloromethane, washed with saturated aqueous ammonium chloride, then with brine, dried (magnesium sulfate) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 24 g RediSep™ silica gel cartridge) eluting with a gradient of 0-60% ethyl acetate in isoheptane gave the title product as a yellow solid (101 mg, 0.2 mmol, 35%) .

LC/MS (C ₂₄ H ₂₈ ClN ₅ O ₃ S) 502 [M+H]+; RT 1.29 (LCMS-V-BI).

1H NMR (400 MHz, DMSO-06) δ 7.77 (d, J=1.2 Hz, 1H), 7.40-7.33 (m, 4H), 7.32-7.26 (m, 1H), 4.56 (s, 2H), 4.25 ( q, J=7.1 Hz, 2H), 3.70 (s, 3H), 3.66-3.56 (m, 1H), 3.45-3.33 (m, 2H), 3.02 (td, J=10.1, 8.7, 3.0 Hz, 2H) , 2.41 (d, J=1.0 Hz, 3H), 2.07-1.93 (m, 2H), 1.79-1.63 (m, 2H), 1.30 (t, J=7.1 Hz, 3H).

Step B: ethyl 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)-5-[4(benzyloxy)piperidin-1 - yl]-1,3-thiazol-4-carboxylate.

A solution of the product from step A (101 mg, 0.2 mmol, 1 eq.), 2-aminobenzothiazole (36.2 mg, 0.24 mmol, 1.2 eq.), ^^diisopropylethylamine (0.1 ml, 0.6 mmol, 3 eq.) and xantphos (11.6 mg, 0.02 mmol, 0.1 eq.) in 1,4-dioxane (4 ml) was bubbled with nitrogen (10 min), then tris(dibenzylideneacetone)dipalladium(0) (9.19 mg, 0.01 mmol, 0.05 eq.) was added, and the mixture heated in a sealed flask at 150°C overnight. The reaction mixture was allowed to cool to ambient temperature and concentrated in vacuo. Purification by reverse phase automated flash chromatography (CombiFlash Rf, C18 13 g RediSep column) eluting with a gradient of 5-95% acetonitrile (pH 4; formic acid) in water (pH 4; formic acid) provided the desired product in the form of a yellow glassy substance (31.2 mg, 0.05 mmol, 25%).

LC/MS (C ₃₁ H ₃₃ N ₇ O ₃ S ₂ ) 616 [M+H] ⁺ ; RT 1.36 (LCMS-V-BI).

1H NMR (400 MHz, DMSO-06) δ 7.90 (s, 1H), 7.63 (br s + s, 2H), 7.43-7.32 (m, 5H), 7.32-7.25 (m, 1H), 7.24-7.16 ( m, 1H), 4.56 (s, 2H), 4.26 (q, J=7.1 Hz, 2H), 3.70 (s, 3H), 3.68-3.57 (m, 1H), 3.46-3.35 (m, 2H), 3.082 .97 (m, 2H), 2.46 (s, 3H), 2.08-1.97 (m, 2H), 1.80-1.69 (m, 2H), 1.32 (t, 3H).

Step C: 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3 -yl} (methyl)amino)-5 -[4(benzyloxy)piperidin-1 -yl ]-1,3-thiazole-4-carboxylic acid.

To a solution of the product from step B (31.2 mg, 0.05 mmol, 1 eq.) in 1,4-dioxane (3 ml) was added lithium hydroxide monohydrate (10.6 mg, 0.25 mmol, 5 eq.), and the mixture was heated at 100°C during the night. The reaction mixture was allowed to cool to ambient temperature and concentrated in vacuo. Purification by reverse phase automated flash chromatography (CombiFlash Rf, C18 13 g RediSep column) eluting with a gradient of 5-95% acetonitrile in water gave the title product as a cream solid (12.2 mg, 0.02 mmol, 41% ) [as lithium salt].

HRMS-ERI (m/z) [M+H] ⁺ , calculated for C ₂₉ H ₃₀ N ₇ O ₃ S ₂ : 588.1846; found: 588.1854.

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Example 76: 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)-5-[3-(2fluoro-4-{3 -[(2-hydroxyethyl)(methyl)amino]prop-1 -yn-1 -yl}phenoxy)propyl] - 1,3-thiazol-4-carboxylic acid

Step A: ethyl 5-(3-{4-[3-({2-[(tert-butyldimethylsilyl)oxy]ethyl}(methyl)amino)prop-1-yn-1-yl]2-fluorophenoxy}propyl) -2-[methyl(5 -methyl-6-{ [(2Z)-3 - {[2-(trimethylsilyl)ethoxy]methyl }-2,3-dihydro1,3-benzothiazol-2-ylidene]amino}pyridazin- 3-yl)amino]-1,3-thiazol-4-carboxylate.

To a solution of synthesis product 5j (747 mg, 0.96 mmol, 1 eq.) in acetonitrile (50 ml) was added (tert-butyldimethylsiloxy)acetaldehyde (367 μl, 1.93 mmol, 2 eq.) followed by the addition of sodium triacetoxyborohydride (612 mg, 2.89 mmol, 3 eq.) and glacial acetic acid (50 μl), and the mixture was stirred at ambient temperature overnight.

The reaction mixture was partitioned between ethyl acetate and saturated aqueous sodium bicarbonate, and the organic phase was dried (magnesium sulfate) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 24 g RediSep™ silica gel cartridge) eluting with a gradient of 0-60% ethyl acetate in isoheptane gave the title product as a clear oil (907 mg, 0.96 mmol, 100%).

LC/MS (C ₄₆ H ₆₄ FN ₇ O ₅ Si ₂ S ₂ ) 934 [M+H]+; RT 1.47 (ZHMS-U-V2).

¹ H NMR (400 MHz, DMSO) δ 7.84 (d, 1H), 7.67 (d, 1H), 7.50-7.40 (m, 2H), 7.31-7.22 (m, 2H), 7.217.12 (m, 2H) , 5.87 (s, 1H), 4.26 (q, J=7.1 Hz, 2H), 4.14 (t, J=6.1 Hz, 2H), 3.77 (s, 3H), 3.75-3.62 (m, 4H), 3.50 ( s, 2H), 2.46 (s, 3H), 2.24 (s, 3H), 2.19-2.06 (m, 2H), 1.29 (t, 3H), 1.24 (s, 6H), 0.96-0.89 (m, 2H) , 0.04 (s, 9H), -0.11 (s, 9H).

Step B: ethyl 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)-5-[3-(2fluoro-4-{ 3-[(2-hydroxyethyl)(methyl)amino]prop-1-yn-1-yl}phenoxy)propyl] - 1,3-thiazol-4-carboxylate.

Trifluoroacetic acid (13 ml) was added to a stirred solution of the product from step A (907 mg, 0.97 mmol, 1 eq.) in dichloromethane (40 ml), and the mixture was stirred at ambient temperature for 9 hours. The reaction mixture was diluted with dichloromethane, cooled to 0°C and made alkaline by adding 2N. aqueous sodium hydroxide solution, and the organic phase was dried (magnesium sulfate) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 12 g RediSep™ silica gel cartridge) eluting with a gradient of 0-20% methanol in dichloromethane gave the title product as a yellow solid (95 mg, 0.14 mmol, 14%).

LC/MS (C ₃₄ H ₃₆ FN ₇ O ₄ S ₂ ) 690 [M+H]+; RT 1.98 (LCMS-VC).

1H NMR (400 MHz, DMSO-dg) δ 7.91 (d, J=7.9 Hz, 1H), 7.67 (s, 1H), 7.56-7.48 (m, 1H), 7.39 (dt, 1H), 7.29 (dd, J=11.9, 1.9 Hz, 1H), 7.26-7.11 (m, 3H), 4.41 (t, J=5.5 Hz, 1H), 4.26 (q, J=7.1 Hz, 2H), 4.14 (t, J=6.1 Hz, 2H), 3.76 (s, 3H), 3.48 (s, 3H), 3.47-3.43 (m, 2H), 3.31-3.25 (m, 2H), 2.46 (s, 3H), 2.24 (s, 3H) , 2.21-2.09 (m, 2H), 1.29 (t, J=7.1 Hz, 3H).

Step C: 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)-5-[3-(2-fluoro4-{3 -[(2-hydroxyethyl)(methyl)amino]prop-1-in-1-yl}phenoxy)propyl]-1,3-thiazol-4-carboxylic acid.

To a solution of the product from step B (95 mg, 0.14 mmol, 1 eq.) in 1,4-dioxane (15 ml) was added lithium hydroxide monohydrate (57.8 mg, 1.38 mmol, 10 eq.), and the mixture was heated at 100°C for 6 hours. The reaction mixture was allowed to cool to ambient temperature and concentrated in vacuo. Purification by reverse phase automated flash chromatography (CombiFlash Rf, C18 13 g RediSep column) eluting with a gradient of 5-95% acetonitrile (pH 9; ammonium acetate) in water (pH 9; ammonium acetate) gave the desired product as a yellow solid (61.8 mg, 0.09 mmol, 67%).

MSHR-ERI (m/z) [M+H]+, calculated for C ₃₂ H ₃₃ FN ₇ O ₄ S ₂ : 662.2014; found: 662.2035.

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Example 77: 2-([6-[('1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)-5-[(^)-3phenoxypyrrolidine-1 - yl]-1,3-thiazole-4-carboxylic acid

Step A: ethyl 2-[methyl(5-methyl-6-{[(27)-3-{[2-(trimethylsilyl)ethoxy]methyl}-2,3-dihydro-1,3benzothiazol-2-ylidene]amino } pyridazin-3 -yl)amino]-5-[(3 S)-3-phenoxypyrrolidin-1 -yl]-1,3-thiazol-4carboxylate.

Synthesis product 11b (200 mg, 0.31 mmol, 1 equiv.), (3S)-3-phenoxypyrrolidine hydrochloride (0.09 ml, 0.63 mmol, 2 equiv.), cesium carbonate (308 mg, 0.94 mmol) was loaded into a vessel dried in an oven. , 3 equiv.) and 1,4-dioxane (8 ml). The mixture was bubbled with nitrogen (10 min), then rac-BINAP Pd G3 (31.2 mg, 0.03 mmol, 0.1 eq.) was added and the mixture was heated at 100°C for 6 h.

The reaction mixture was diluted with ethyl acetate, then washed with water followed by brine, dried (magnesium sulfate) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 12 g RediSep™ silica gel cartridge) eluting with a gradient of 0-50% ethyl acetate in isoheptane gave the title product as a yellow foam (184 mg, 0.26 mmol, 81%).

LC/MS (C ₃₅ H ₄₃ N ₇ O ₄ SiS ₂ ) 718 [M+H]+; RT 1.54 (ZHMS-U-B1).

¹ H NMR (400 MHz, DMSO-dg) δ 7.82 (d, 1H), 7.58 (d, 1H), 7.48-7.37 (m, 2H), 7.35-7.20 (m, 4H), 7.016.91 (m, 3H), 5.86 (s, 2H), 5.18-5.13 (m, 1H), 4.21 (qd, J=7.1, 2.3 Hz, 2H), 3.96 (dd, J=11.8, 4.5 Hz, 1H), 3.773.60 (m, 2H), 3.68 (s, 3H), 3.67-3.61 (m, 1H), 3.49 (td, J=8.7, 2.9 Hz, 1H), 3.37-3.32 (m, 1H), 2.45 (d, J =1.0 Hz, 3H), 2.37-2.26 (m, 1H), 2.25-2.15 (m 1H), 1.27 (t, J=7.1 Hz, 3H), 0.92 (dd, J=8.4, 7.6 Hz, 2H), 0.00 (s, 9H).

Step B: ethyl 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)-5-[(3S)3-phenoxypyrrolidin-1 -yl]-1,3-thiazol-4-carboxylate.

Hydrochloric acid (4 M in 1,4-dioxane; 3 mL, 12 mmol, 47 eq.) was added to a stirred solution of the product from step A (184 mg, 0.26 mmol, 1 eq.) in 1,4-dioxane (7 mL ), and the mixture was stirred at 50°C overnight. The reaction mixture was diluted with dichloromethane, washed with saturated aqueous sodium bicarbonate, then with brine, dried (magnesium sulfate) and concentrated in vacuo. Purification by reverse phase automated flash chromatography (CombiFlash Rf, C18 13 g RediSep column) eluting with 5-95% acetonitrile (pH4; formic acid) in water (pH 4; formic acid) gave the desired product as a yellow gum ( 55.8 mg, 0.09 mmol, 37%).

LC/MS (C ₂₉ H ₂₉ N ₇ O ₃ S ₂ ) 588 [M+H]+; RT 1.32 (ZHMS-U-B1).

¹ H NMR (400 MHz, DMSO-dg) δ 7.89 (br s, 1H), 7.59 (br s + s, 2H), 7.38 (t, J=7.5 Hz, 1H), 7.33-7.26 (m, 2H) , 7.19 (t, J=7.6 Hz, 1H), 7.01-6.91 (m, 3H), 5.18-5.13 (m, 1H), 4.21 (qd, J=7.1, 2.3 Hz, 2H), 3.96 (dd, J =11.8, 4.5 Hz, 1H), 3.66 (s, 3H), 3.66-3.59 (m, 1H), 3.48 (td, J=9.2, 8.8, 2.8 Hz, 1H), 3.38-3.28 (m, 1H), 2.44 (s, 3H), 2.40-2.26 (m, 1H), 2.24-2.15 (m, 2H), 1.26 (t, J=7.1 Hz, 3H).

Step C: 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)-5-[(3S)-3phenoxypyrrolidin-1 -yl ] -1,3-thiazole-4-carboxylic acid.

To a solution of the product from step B (55.8 mg, 0.09 mmol, 1 eq.) in 1,4-dioxane (5 ml) was added lithium hydroxide monohydrate (19.9 mg, 0.47 mmol, 5 eq.), and the mixture was heated at 100°C during the night. The reaction mixture was allowed to cool to ambient temperature, concentrated in vacuo, and the residue triturated with water, filtered and dried in vacuo to give the title product as a yellow solid (38.5 mg, 0.07 mmol, 72%) [as lithium salt].

HRMS-ERI (m/z) [M+H]+, calculated for C ₂₇ H ₂₆ N ₇ O ₃ S ₂ : 560.1533; found: 560.1541.

Example 78: 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)-5cyclobutyl-1,3-thiazol-4-carboxylic acid

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Step A: ethyl 5-cyclobutyl-2-[methyl(5-methyl-6-{[(2Z)-3-{[2-(trimethylsilyl)ethoxy]methyl}-2,3dihydro-1,3-benzothiazol-2 -ylidene]amino}pyridazin-3-yl)amino]-1,3-thiazol-4-carboxylate.

The oven-dried flask was charged with synthesis product 11b (500 mg, 0.79 mmol, 1 eq.) and medip iodide (300 mg, 1.57 mmol, 2 eq.), and the flask was evacuated and purged with nitrogen (x3), and then dimethyl acetamide was added (15 ml). Cyclobutylzinc bromide (0.5 M in tetrahydrofuran; 9.44 mL, 4.72 mmol, 6 eq.) was added in one portion and the reaction mixture was stirred overnight at ambient temperature. The mixture was diluted with dichloromethane and then quenched with saturated aqueous sodium bicarbonate, causing a precipitate to form. The mixture was stirred vigorously for 5 minutes, then filtered through a dichloromethane-soaked celite pad, and eluted with dichloromethane and saturated aqueous sodium bicarbonate. The layers were separated and the aqueous phase was extracted with dichloromethane (x2). The combined organic extracts were washed with brine, dried (magnesium sulfate) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 24 g RediSep™ silica gel cartridge) eluting with a gradient of 0-30% ethyl acetate in isoheptane gave the title product as a pale yellow glass/foam (313 mg, 0.51 mmol, 65%).

LC/MS (C ₂₉ H ₃₈ N ₆ O ₃ SiS ₂ ) 611 [M+H]+; RT 1.38 (ZHMS-U-V2).

1H NMR (400 MHz, CDCl ₃ ) δ 7.60 (dt, J=7.5, 0.9 Hz, 1H), 7.45-7.32 (m, 3H), 7.21 (ddd, J=7.6, 5.9, 2.5 Hz, 1H), 5.85 (s, 2H), 4.51-4.28 (m, 3H), 3.84 (s, 3H), 3.81-3.67 (m, 2H), 2.62-2.47 (m, 2H), 2.46 (s, 3H), 2.382.16 (m, 2H), 2.15-1.83 (m, 2H), 1.42 (t, J=7.1 Hz, 3H), 1.05-0.93 (m, 2H), -0.07 (s, 9H).

Step B: 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl} (methyl)amino)-5 cyclobutyl-1,3-thiazol-4-carboxylic acid.

Trifluoroacetic acid (5.04 mL, 65.8 mmol, 130 eq.) was added to a stirred solution of the product from Step A (309 mg, 0.51 mmol, 1 eq.) in dichloromethane (10 mL), and the mixture was stirred at ambient temperature overnight. The reaction mixture was concentrated in vacuo. While stirring, 1,4-dioxane (10 ml) was added to obtain a viscous off-white suspension, to which 1N was added. aqueous sodium hydroxide solution (5 ml), and the mixture was refluxed for 20 minutes. The mixture was allowed to cool to ambient temperature, then filtered through a pad of celite, eluting with water and then dioxane, and concentrated in vacuo. The residue was partitioned between chloroform and saturated aqueous sodium bicarbonate, and the organic phase was dried (magnesium sulfate) and concentrated in vacuo. The solid was triturated with diethyl ether, filtered and dried in vacuo to give the title product as a yellow powder (98.5 mg, 0.22 mmol, 43%).

HRMS-ERI (m/z) [M+H]+, calculated for C ₂₁ H ₂₁ N ₆ O ₂ S ₂ : 453.1162; found: 453.1172.

Example 79: 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)-5-[(3R)-3phenoxypyrrolidin-1 -yl ]-1,3-thiazole-4-carboxylic acid

Step A: ethyl 2-[methyl(5-methyl-6-{[(2Z)-3-{[2-(trimethylsilyl)ethoxy]methyl}-2,3-dihydro-1,3benzothiazol-2-ylidene] amino } pyridazin-3-yl)amino]-5-[(3R)-3-phenoxypyrrolidin-1-yl]-1,3-thiazol-4carboxylate.

Synthesis product 11b (300 mg, 0.44 mmol, 1 equiv.), ^)-3-phenoxypyrrolidine (0.13 ml, 0.88 mmol, 2 equiv.), cesium carbonate (430 mg, 1.32 mmol, 3 eq.) and 1,4-dioxane (12 ml). The mixture was bubbled with nitrogen (10 min), rac-BINAP Pd G3 (43.6 mg, 0.04 mmol, 0.1 eq.) was added and heated at 100°C overnight. The reaction mixture was diluted with ethyl acetate, then washed with water followed by brine, dried (magnesium sulfate) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 12 g RediSep™ silica gel cartridge) eluting with a gradient of 0-50% ethyl acetate in isoheptane gave the title product as a yellow foam (253 mg, 0.35 mmol, 80%).

LC/MS (C ₃₅ H4 ₃ N7O ₄ SiS ₂ ) 718 [M+H]+; RT 1.54 (LCMS-V-BI).

1H NMR (400 MHz, DMSO-c1 ₆ ) δ 7.85-7.78 (m, 1H), 7.57 (d, J=1.1 Hz, 1H), 7.47-7.38 (m, 2H), 7.35-7.19 (m, 3H) , 7.01-6.80 (m, 3H), 5.85 (s, 2H), 5.22-5.10 (m, 1H), 4.21 (qd, J=7.1, 2.4 Hz, 2H), 3.96 (dd, J=11.8, 4.5 Hz , 1H), 3.76-3.59 (m, 3H), 3.49 (td, J=8.6, 2.8 Hz, 1H), 3.33 (s, 4H), 3.36-3.28 (m, 1H), 2.44 (d, J=1.0 Hz, 3H), 2.39-2.28 (m, 1H), 2.26-2.16 (m, 1H), 1.27 (t, J=7.1 Hz, 3H), 0.96-0.83 (m, 2H), -0.11 (s, 9H ).

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Step B: ethyl 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)-5-[(3R)3-phenoxypyrrolidin-1 -yl]-1,3-thiazol-4-carboxylate.

Hydrochloric acid (4 M in 1,4-dioxane; 4 mL, 16 mmol, 45 eq.) was added to a stirred solution of the product from step A (253 mg, 0.35 mmol, 1 eq.) in 1,4-dioxane (8 mL ), and the mixture was stirred at 50°C overnight. The reaction mixture was diluted with dichloromethane, washed with saturated aqueous sodium bicarbonate, then with brine, dried (magnesium sulfate) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 12 g RediSep™ silica gel cartridge) eluting with a gradient of 0-5% methanol in dichloromethane gave the title product as a yellow foam (189 mg, 0.32 mmol, 91%).

LC/MS (C ₂₉ H ₂₉ N ₇ O ₃ S ₂ ) 588 [M+H]+; RT 1.32 (LCMS-V-BI).

1H NMR (400 MHz, DMSO-de) δ 10.98 (br s, 1H), 7.94-7.87 (m, 1H), 7.60 (br s + s, 2H), 7.39 (t, J=7.7 Hz, 1H), 7.34-7.27 (m, 2H), 7.25-7.16 (s, 1H), 7.02-6.87 (m, 3H), 5.20-5.13 (m, 1h), 4.21 (qd, J=7.1, 2.2 Hz, 2H), 3.97 (dd, J=11.8, 4.5 Hz, 1H), 3.67 (s, 3H), 3.74-3.60 (m, 1H), 3.55-3.43 (m, 1H), 3.38-3.30 (m, 1H), 2.45 ( s, 3H), 2.41-2.27 (m, 1H), 2.26-2.17 (m, 1H), 1.27 (t, J=7.1 Hz, 3H).

Step C: 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)-5-[(3R)-3phenoxypyrrolidin-1 -yl ]-1,3-thiazole-4-carboxylic acid.

To a solution of the product from step B (189 mg, 0.32 mmol, 1 eq.) in 1,4-dioxane (10 ml) was added lithium hydroxide monohydrate (67.4 mg, 1.61 mmol, 5 eq.) and the mixture was refluxed at during the night. The reaction mixture was allowed to cool to ambient temperature, then concentrated in vacuo, triturated with water, filtered and dried in vacuo to give the title product as a beige solid (118 mg, 0.21 mmol, 65%) [as lithium salt].

HRMS-ERI (m/z) [M+H]+, calculated for C ₂₇ H ₂₆ N ₇ O ₃ S ₂ : 560.1533; found: 560.1535.

Example 80: 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)-5(piperidin-1-yl)-1,3 -thiazole-4-carboxylic acid hr s

Step A: ethyl 5-(piperidin-1-yl)-1,3-thiazol-4-carboxylate.

A solution of ethyl 5-bromothiazole-4-carboxylate (500 mg, 2.12 mmol, 1 eq.), piperidine (0.25 ml, 2.54 mmol, 1.2 eq.) and 1,8-diazabicyclo[5.4.0]undec-7-ene ( 0.64 ml, 4.24 mmol, 2 eq.) in acetonitrile (15 ml) was heated at 80°C overnight. The reaction mixture was allowed to cool to ambient temperature, then concentrated in vacuo and purified by automated flash column chromatography (CombiFlash Rf, 24 g RediSep™ silica gel cartridge) eluting with a gradient of 0-60% ethyl acetate in isoheptane to give the title product as a brown oil (376 mg, 1.57 mmol, 74%).

LC/MS (C11H16N2O2S) 241 [M+H]+; RT 1.12 (LCMS-V-BI).

1H NMR (400 MHz, DMSO^) δ 8.45 (s, 1H), 4.24 (q, J=7.1 Hz, 2H), 3.15-3.07 (m, 4H), 1.67 (p, J=5.6 Hz, 4H), 1.58-1.49 (m, 2H), 1.28 (t, J=7.1 Hz, 3H).

Step B: ethyl 2-bromo-5-(piperidin-1-yl)-1,3-thiazol-4-carboxylate.

N-Bromosuccinimide (453 mg, 2.55 mmol, 1.2 eq.) was added to a stirred solution of the product from step A (510 mg, 2.12 mmol, 1 eq.) in acetonitrile (20 ml), and the mixture was stirred at ambient temperature for 7 h. The reaction mixture was partitioned between 10% aqueous sodium thiosulfate and ethyl acetate, and the organic phase was washed with water followed by brine, dried (magnesium sulfate) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 24 g RediSep™ silica gel cartridge) eluting with a gradient of 0-40% ethyl acetate in isoheptane gave the title product as a colorless oil (482 mg, 1.51 mmol, 71%).

LC/MS (C ₁₁ H ₁₅ BrN ₂ O ₂ S) 273 [M-OEf] ⁺ ; RT 1.14 (LCMS-V-BI).

¹ HNMR (400 MHz, DMSO-deO δ 4.23 (q, J=7.1 Hz, 2H), 3.18-3.10 (m, 4H), 1.70-1.60 (m, 4H), 1.59-1.49 (m, 2H), 1.28 (t, J=7.1 Hz, 3H).

Step C: ethyl 2-{[(tert-butoxy)carbonyl](methyl)amino}-5-(piperidin-1-yl)-1,3-thiazol-4carboxylate.

The product from step B (482 mg, 1.51 mmol, 1 eq.) and tert-butyl methyl carbamate (0.26 ml, 1.81 mmol, 1.2 eq.) were added under nitrogen to a solution of tris(dibenzylideneacetone)dipalladium(0) (69.1 mg, 0.08 mmol, 0.05 eq.) and xantphos (87.4 mg, 0.15 mmol, 0.1 eq.) in 1,4-dioxane (10 ml). Cesium carbonate (738 mg, 2.26 mmol, 1.5 eq.) was added and the mixture was heated at 100°C for 7.5 hours. The solution was allowed to cool to ambient temperature and concentrated in vacuo. The mixture was diluted with ethyl acetate

- 154 046336 volume and washed with water followed by brine, dried (magnesium sulfate) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 50 g RediSep™ silica gel cartridge) eluting with a gradient of 0-50% ethyl acetate in isoheptane gave the title product as a yellow gum (434 mg, 1.17 mmol, 78%).

LC/MS (C ₁₇ H ₂₇ N ₃ O ₄ S) 370 [M+H]+; RT 1.304 (ZHMS-U-B1).

1H NMR (400 MHz, DMSO-dg) δ 4.21 (q, J=7.1 Hz, 2H), 3.37 (s, 3H), 3.11-3.03 (m, 4H), 1.64 (p, J=5.6 Hz, 4H) , 1.55-1.51 (m, 2H), 1.51 (s, 9H), 1.26 (t, J=7.1 Hz, 3H).

Step D: Ethyl 2-(methylamino)-5-(piperidin-1-yl)-1,3-thiazol-4-carboxylate.

Trifluoroacetic acid (1.32 ml, 17.6 mmol, 15 eq.) at 0°C was added to a stirred solution of the product from step C (434 mg, 1.17 mmol, 1 eq.) in dichloromethane (15 ml), and the mixture was allowed to warm to ambient temperature medium and stirred overnight. The reaction mixture was partitioned between saturated aqueous sodium bicarbonate and dichloromethane, and the organic phase was washed with brine, dried (magnesium sulfate) and concentrated in vacuo. The resulting solid was triturated with diethyl ether, collected by filtration and dried in vacuo to give the title product as a white solid (197 mg, 0.73 mmol, 62%).

LC/MS (C ₁₂ H ₁₉ N ₃ O ₂ S) 270 [M+H]+; RT 0.91 (ZhKhMS-U-B1).

1H NMR (400 MHz, DMSOO δ 7.26 (q, J=4.8 Hz, 1H), 4.17 (q, J=7.1 Hz, 2H), 2.91-2.84 (m, 4H), 2.75 (d, J=4.8 Hz, 3H), 1.61 (p, J=5.6 Hz, 4H), 1.52-1.42 (m, 2H), 1.25 (t, J=7.1 Hz, 3H).

Step E: ethyl 2-[(6-chloro-5-methylpyridazin-3-yl)(methyl)amino]-5-(piperidin-1-yl)-1,3-thiazol-4carboxylate.

Sodium hydride (60% in mineral oil; 35.2 mg, 0.88 mmol, 1.2 eq.) was added to a cooled solution of the product from step D (197 mg, 0.73 mmol, 1 eq.) and 3,6-dichloro-4-methylpyridazine (119 mg, 0.73 mmol, 1 eq.) in tetrahydrofuran (8 ml), and the mixture was allowed to warm to ambient temperature and stirred for 3.5 hours. The reaction mixture was diluted with dichloromethane, washed with saturated aqueous ammonium chloride followed by brine, dried ( magnesium sulfate) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 24 g RediSep™ silica gel cartridge) eluting with a gradient of 0-60% ethyl acetate in isoheptane provided the title product as a yellow solid (131 mg, 0.33 mmol, 45%).

LC/MS (C ₁₇ H ₂₂ ClN ₅ O ₂ S) 396 [M+H]+; RT 1.20 (ZHMS-U-B1).

1H NMR (400 MHz, DMSOO δ 7.76 (d, J=1.0 Hz, 1H), 4.25 (q, J=7.1 Hz, 2H), 3.70 (s, 3H), 3.16-3.08 (m, 4H), 2.41 ( d, J=1.0 Hz, 3H), 1.66 (q, J=5.8 Hz, 4H), 1.60-1.46 (m, 2H), 1.30 (t, J=7.1 Hz, 3H).

Step F: ethyl 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)-5(piperidin-1-yl)-1, 3-thiazole-4-carboxylate.

A solution of the product from step E (131 mg, 0.33 mmol, 1 eq.), 2-aminobenzothiazole (74.7 mg, 0.5 mmol, 1.5 eq.), N,N-diisopropylethylamine (0.16 ml, 0.99 mmol, 3 eq.) and xantphos (19.2 mg, 0.03 mmol, 0.1 eq.) in 1,4-dioxane (7 ml) was bubbled with nitrogen (10 min), then tris(dibenzylideneacetone)dipalladium(0) (15.2 mg, 0.02 mmol, 0.05 eq.) was added. and the mixture was heated in a stoppered flask at 150°C overnight. The reaction mixture was allowed to cool to ambient temperature, then partitioned between ethyl acetate and water, and the organic phase was washed with brine, dried (magnesium sulfate) and concentrated in vacuo. Purification by reverse phase automated flash chromatography (CombiFlash Rf, C18 13 g RediSep column) eluting with a gradient of 5-95% acetonitrile (pH4; formic acid) in water (pH 4; formic acid) gave the desired product as yellow foam (64.8 mg, 0.13 mmol, 38%).

LC/MS (C ₂₄ H ₂₇ N ₇ O ₂ S ₂ ) 510 [M+H]+; RT 1.32 (ZHMS-U-B1).

1H NMR (400 MHz, DMSOHO δ 7.91 (br s, 1H), 7.63 (br s + s, 2H), 7.39 (t, J=7.6 Hz, 1H), 7.22 (t, J=7.5 Hz, 1H), 4.26 (q, 2H), 3.70 (s, 3H), 3.18-3.08 (m, 4H), 2.46 (s, 3H), 1.72-1.61 (m, 4H), 1.59-1.47 (m, 2H), 1.31 ( t, J=7.1, 2.7 Hz, 3H).

Step G: 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)-5 (piperidin-1-yl)-1,3 -thiazole-4-carboxylic acid.

To a solution of the product from step F (64.8 mg, 0.13 mmol, 1 eq.) in 1,4-dioxane (3 ml) was added lithium hydroxide monohydrate (26.7 mg, 0.64 mmol, 5 eq.) and the mixture was refluxed at during the night. The reaction mixture was allowed to cool to ambient temperature and concentrated in vacuo. Purification by reverse phase automated flash chromatography (CombiFlash Rf, C18 13 g RediSep column) eluting with a gradient of 5-95% acetonitrile in water gave the title product as a yellow solid (23 mg, 0.05 mmol, 38% ).

HRMS-ERI (m/z) [M+H] ⁺ , calculated for C ₂₂ H ₂₄ N ₇ O ₂ S ₂ : 482.1427; found: 482.1435.

- 155 046336

Example 81: 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)-5-(4phenoxypiperidin-1 -yl)-1, 3-thiazole-4-carboxylic acid

Step A: ethyl 5-(4-phenoxypiperidin-1-yl)-1,3-thiazol-4-carboxylate.

A solution of ethyl 5-bromothiazole-4-carboxylate (500 mg, 2.12 mmol, 1 eq.), 4-phenoxypiperidine (451 mg, 2.54 mmol, 1.2 eq.) and 1,8-diazabicyclo[5.4.0]undec-7- ene (0.64 ml, 4.24 mmol, 2 eq.) in acetonitrile (15 ml) was heated at 80°C overnight, then allowed to cool to ambient temperature and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 40 g RediSep™ silica gel cartridge) eluting with a gradient of 0-60% ethyl acetate in isoheptane gave the title product as a brown oil (424 mg, 1.27 mmol, 60%).

LC/MS (C ₁₇ H ₂₀ N ₂ O ₃ S) 333 [M+H]+; RT 1.28 (LCMS-V-BI).

¹ H NMR (400 MHz, DMSO-de) δ 8.49 (s, 1H), 7.35-7.25 (m, 2H), 7.05-6.96 (m, 2H), 6.94 (tt, J=7.3, 1.0 Hz, 1H) , 4.61 (tt, J=7.6, 3.6 Hz, 1H), 4.25 (q, J=7.1 Hz, 2H), 3.45-3.35 (m, 2H), 3.20-3.08 (m, 2H), 2.15-2.02 (m , 2H), 1.88-1.75 (m, 2H), 1.29 (t, J=7.1 Hz, 3H).

Step B: ethyl 2-bromo-5-(4-phenoxypiperidin-1-yl)-1,3-thiazol-4-carboxylate.

N-Bromosuccinimide (272 mg, 1.53 mmol, 1.2 eq.) was added to a stirred solution of the product from Step A (424 mg, 1.27 mmol, 1 eq.) in acetonitrile (20 ml), and the mixture was stirred at ambient temperature for 5.5 h. The reaction mixture was quenched by adding 10% aqueous sodium thiosulfate, then extracted with ethyl acetate, washed with water followed by brine, dried (magnesium sulfate) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 12 g RediSep™ silica gel cartridge) eluting with a gradient of 0-40% ethyl acetate in isoheptane provided the title product as a white solid (404 mg, 0.98 mmol, 77%) .

LC/MS (C ₁₇ H ₁₉ BrN ₂ O ₃ S) 413 [M+H]+; RT 1.26 (LCMS-V-BI).

1H NMR (400 MHz, DMSO^) δ 7.35-7.25 (m, 2H), 7.04-6.89 (m, 3H), 4.62 (dt, J=7.6, 3.9 Hz, 1H), 4.25 (q, 2H), 3.46 -3.36 (m, 2H), 3.24-3.13 (m, 2H), 2.12-2.01 (m, 2H), 1.87-1.74 (m, 2H), 1.28 (t, 3H).

Step C: ethyl 2-{[(tert-butoxy)carbonyl](methyl)amino}-5-(4-phenoxypiperidin-1-yl)-1,3thiazol-4-carboxylate.

The product from step B (404 mg, 0.98 mmol, 1 eq.) and tert-butyl methyl carbamate (0.17 ml, 1.18 mmol, 1.2 eq.) were added under a nitrogen atmosphere to a solution of tris(dibenzylideneacetone)dipalladium(0) (45 mg, 0.05 mmol, 0.05 eq.) and xantphos (56.8 mg, 0.1 mmol, 0.1 eq.) in 1,4-dioxane (10 ml). Cesium carbonate (480 mg, 1.47 mmol, 1.5 eq) was added and the mixture was heated at 100°C overnight. The reaction mixture was allowed to cool to ambient temperature and then concentrated in vacuo. The residue was partitioned between ethyl acetate and water, and the organic phase was washed with brine, dried (magnesium sulfate) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 24 g RediSep™ silica gel cartridge) eluting with a gradient of 0-50% ethyl acetate in isoheptane gave the title product as a yellow gum (285 mg, 0.62 mmol, 63%).

LC/MS (C ₂₃ H ₃₁ N ₃ O ₅ S) 462 [M+H]+; RT 1.37 (LCMS-V-BI).

1H NMR (400 MHz, DMSO^ ₆ ) δ 7.34-7.24 (m, 2H), 7.02-6.97 (m, 2H), 6.93 (tt, J=7.2, 1.1 Hz, 1H), 4.58 (dt, J=7.9 , 4.1 Hz, 1H), 4.23 (q, J=7.1 Hz, 2H), 3.38 (s, 3H), 3.37-3.33 (m, 2H), 3.09 (ddd, J=11.8, 8.5, 3.2 Hz, 2H) , 2.15-1.99 (m, 2H), 1.79 (dtd, J=12.3, 8.2, 3.6 Hz, 2H), 1.52 (s, 9H), 1.27 (t, J=7.1 Hz, 3H).

Step D: Ethyl 2-(methylamino)-5-(4-phenoxypiperidin-1-yl)-1,3-thiazol-4-carboxylate.

Trifluoroacetic acid (0.46 ml, 6.17 mmol, 10 eq.) at 0°C was added to a stirred solution of the product from step C (285 mg, 0.62 mmol, 1 eq.) in dichloromethane (8 ml), and the mixture was allowed to warm to ambient temperature medium and stirred overnight. The reaction mixture was quenched by adding saturated aqueous sodium bicarbonate, extracted with dichloromethane, and the organic extract was washed with brine, dried (magnesium sulfate) and concentrated in vacuo. The resulting solid was triturated with diethyl ether/heptane, filtered and dried in vacuo to give the title product as a cream solid (161 mg, 0.45 mmol, 72%).

LC/MS (C ₁₈ H ₂₃ N ₃ O ₃ S) 362 [M+H]+; RT 1.08 (LCMS-V-BI).

1H NMR (400 MHz, DMSO-d^ δ 7.35-7.24 (m, 3H), 7.02-6.97 (m, 2H), 6.93 (tt, J=7.3, 1.1 Hz, 1H), 4.53

- 156 046336 (dt, J=7.9, 4.1 Hz, 1H), 4.19 (q, J=7.1 Hz, 2H), 3.22-3.12 (m, 2H), 2.90 (ddd, J=11.6, 8.6, 3.2 Hz, 2H), 2.76 (d, J=4.7 Hz, 3H), 2.10-1.95 (m, 2H), 1.77 (dtd, J=12.3, 8.2, 3.6 Hz, 2H), 1.26 (t, J=7.1 Hz, 3H ).

Step E: ethyl 2-[(6-chloro-5-methylpyridazin-3-yl)(methyl)amino]-5-(4-phenoxypiperidin-1-yl)-1,3thiazol-4-carboxylate.

Sodium hydride (60% in mineral oil; 21.4 mg, 0.53 mmol, 1.2 eq.) was added to a cooled solution of the product from step D (161 mg, 0.45 mmol, 1 eq.) and 3,6-dichloro-4-methylpyridazine (72.6 mg, 0.45 mmol, 1 eq.) in tetrahydrofuran (5 ml), and the mixture was allowed to warm to ambient temperature and stirred overnight. The reaction mixture was diluted with dichloromethane, then washed with saturated aqueous ammonium chloride followed by brine, dried (magnesium sulfate) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 12 g RediSep™ silica gel cartridge) eluting with a gradient of 0-60% ethyl acetate in isoheptane gave the title product as a yellow foam (63.3 mg, 0.13 mmol, 29%).

LC/MS (C ₂₃ H ₂₆ ClN ₅ O ₃ S) 488 [M+H]+; RT 1.289 (LCMS-V-BI).

¹ H NMR (400 MHz, DMSO-de) δ 7.77 (d, J=1.0 Hz, 1H), 7.34-7.25 (m, 2H), 7.04-6.98 (m, 2H), 6.93 (tt, J=7.4, 1.1 Hz, 1H), 4.59 (dt, J=8.1, 4.1 Hz, 1H), 4.25 (q, J=7.1 Hz, 2H), 3.70 (s, 3H), 3.45-3.35 (m, 2H), 3.13 ( ddd, J=11.5, 8.5, 3.2 Hz, 2H), 2.41 (d, J=1.0 Hz, 3H), 2.14-2.01 (m, 2H), 1.81 (dtd, J=12.2, 8.3, 3.5 Hz, 2H) , 1.30 (t, J=7.1 Hz, 3H).

Step F: ethyl 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)-5-(4phenoxypiperidin-1-yl)-1 ,3-thiazole-4-carboxylate.

A solution of the product from step E (63.3 mg, 0.13 mmol, 1 eq.), 2-aminobenzothiazole (29.2 mg, 0.19 mmol, 1.5 eq.), X,X-diisopropylethylamine (0.06 ml, 0.39 mmol, 3 eq.) and xantphos (7.51 mg, 0.01 mmol, 0.1 eq.) in 1,4-dioxane (4 ml) was bubbled with nitrogen (10 min), then tris(dibenzylideneacetone)dipalladium(0) (5.94 mg, 0.01 mmol, 0.05 eq.) was added. and the mixture was heated in a stoppered flask at 150°C overnight. The reaction mixture was allowed to cool to ambient temperature, then partitioned between ethyl acetate and water, and the organic phase was washed with brine, dried (magnesium sulfate) and concentrated in vacuo. Purification by reverse phase automated flash chromatography (CombiFlash Rf, C18 13 g RediSep column) eluting with a gradient of 5-95% acetonitrile (pH 4; formic acid) in water (pH 4; formic acid) gave the desired product as yellow foam (23 mg, 0.04 mmol, 30%).

LC/MS (C ₃₀ H ₃₁ N ₇ O ₃ S ₂ ) 602 [M+H]+; RT 1.37 (LCMS-V-BI).

1H NMR (400 MHz, DMSO-de) δ 7.92 (br s, 1H), 7.64 (br s + s, 2H), 7.36 (dd, J=31.4, 7.1 Hz, 1H), 7.29 (dd, J=8.6 , 7.3 Hz, 2H), 7.25-7.17 (m, 1H), 7.07-6.98 (m, 2H), 6.97-6.89 (m, 1H), 4.61 (dt, 1H), 4.27 (q, 2H), 3.71 ( s, 3H), 3.49-3.36 (m, 2H), 3.22-3.09 (m, 3H), 2.46 (s, 3H), 2.18-2.02 (m, 2H), 1.91-1.73 (m, 2H), 1.32 ( t, 3H).

Step G: 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl} (methyl)amino)-5-(4phenoxypiperidin-1-yl)-1, 3-thiazole-4-carboxylic acid.

To a solution of the product from step F (23 mg, 0.04 mmol, 1 eq.) in 1,4-dioxane (3 ml) was added lithium hydroxide monohydrate (8.02 mg, 0.19 mmol, 5 eq.), and the mixture was refluxed at overnight, then allowed to cool to ambient temperature and concentrated in vacuo. Purification by reverse phase automated flash chromatography (CombiFlash Rf, C18 13 g RediSep column) eluting with a gradient of 5-95% acetonitrile in water gave the title product as a yellow solid (3.7 mg, 0.01 mmol, 17% ).

HRMS-ERI (m/z) [M+H] ⁺ , calculated for C ₂₈ H ₂₈ N ₇ O ₃ S ₂ : 574.1690; found: 574.1697.

Example 82: 2-({6-[( 1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl} (methyl)amino)-5-{3-[4-(3{[ (tert-butoxy)carbonyl] (methyl)amino} prop-1-yn-1 -yl)-2-fluorophenoxy]propyl}-1,3-thiazole-4carboxylic acid

Step A: ethyl 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)-5-(3iodopropyl)-1,3-thiazole -4-carboxylate.

Trifluoroacetic acid (5 ml) was added to a solution of synthesis product 5g (176 mg, 0.24 mmol, 1 eq.) in dichloromethane (5 ml), and the mixture was stirred at ambient temperature for 4 hours. The reaction mixture was diluted with dichloromethane, washed with 2 N . aqueous sodium hydroxide solution,

- 157 046336 dried (magnesium sulfate) and concentrated in vacuo. The residue was triturated with acetonitrile, filtered and dried in vacuo to give the title product as a white solid (93 mg, 0.16 mmol, 64%).

LC/MS (C ₂₂ H ₂₃ IN ₆ O ₂ S ₂ ) 595 [M+H]+; RT 2.63 (LCMS-VC).

¹ H NMR (400 MHz, DMSO-dii) δ 7.92 (s, 1H), 7.69 (s, 1H), 7.54 (br s, 1H), 7.40 (t, J=7.8 Hz, 1H), 7.22 (t, J=7.5 Hz, 1H), 4.31 (q, J=7.1 Hz, 2H), 3.77 (s, 3H), 3.35 (t, J=6.8 Hz, 2H), 3.24-3.16 (m, 2H), 2.47 ( s, 3H), 2.202.11 (m, 2H), 1.34 (t, J=7.1 Hz, 3H).

Step B: ethyl 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)-5-{3-[4(3-{ [(tert-butoxy)carbonyl](methyl)amino}prop-1-yn-1-yl)-2-fluorophenoxy]propyl}-1,3-thiazol-4carboxylate.

To a solution of the product from step A (93 mg, 0.16 mmol, 1 eq.) in dimethylformamide (5 ml) was added the product of synthesis 6a (56.8 mg, 0.2 mmol, 1.3 eq.) and cesium carbonate (153 mg, 0.47 mmol, 3 eq. .), and the mixture was heated at 80°C overnight, then allowed to cool to ambient temperature and concentrated in vacuo. The residue was partitioned between ethyl acetate and brine, and the organic phase was dried (magnesium sulfate) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 12 g RediSep™ silica gel cartridge) eluting with a gradient of 0-100% ethyl acetate in isoheptane gave the title product as a yellow gum (45 mg, 0.06 mmol, 39%).

LC/MS (C ₃₇ H ₄₀ FN ₇ O ₅ S ₂ ) 747 [M+H]+; RT 2.77 (LCMS-VC).

¹ H NMR (400 MHz, DMSO-dii) δ 7.91 (d, J=19.8 Hz, 1H), 7.68 (s, 1H), 7.54 (br s, 1H), 7.39 (t, 1H), 7.33 (d, 1H), 7.26-7.12 (m, 3H), 4.26 (q, 2H), 4.21 (s, 2H), 4.15 (t, J=6.1 Hz, 2H), 3.77 (s, 2H), 3.31-3.22 (m , 2H), 2.85 (s, 3H), 2.47 (s, 3H), 2.20-2.09 (m, 2H), 1.41 (s, 9H), 1.30 (t, 3H).

Step C: 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl} (methyl)amino)-5 -{3-[4-(3{[ (tert-butoxy)carbonyl](methyl)amino}prop-1-yn-1-yl)-2-fluorophenoxy]propyl}-1,3-thiazole-4carboxylic acid.

To a solution of the product from step B (45 mg, 0.06 mmol, 1 eq.) in 1,4-dioxane (5 ml) was added lithium hydroxide monohydrate (25.3 mg, 0.6 mmol, 10 eq.), and the mixture was refluxed at during the night. The reaction mixture was allowed to cool to ambient temperature and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 4 g RediSep™ silica gel cartridge) eluting with a gradient of 0-10% methanol in dichloromethane provided the title product as a yellow solid (5.1 mg, 0.01 mmol, 12%) .

HRMS-ERI (m/z) [M+H]+, calculated for C ₃₅ H ₃₇ FN ₇ O ₅ S ₂ : 718.2276; found: 718.2284.

Example 83: 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)-5(piperidin-4-yl)-1,3 -thiazole-4-carboxylic acid

Step A: ethyl 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)-5(piperidin-4-yl)-1, 3-thiazole-4-carboxylate.

Trifluoroacetic acid (1 mL, 13.4 mmol, 166 eq.) was added to a stirred solution of Synthesis 5k, Step B (60 mg, 0.08 mmol, 1 eq.) in dichloromethane (4 mL), and the mixture was stirred at ambient temperature for nights. The reaction mixture was loaded onto an SCX-2 cartridge (5 g, preconditioned with dichloromethane then methanol) and then washed with dichloromethane followed by a wash with methanol. Performing elution 3.5N. methanolic ammonia and concentration in vacuo afforded the title product as a yellow solid (40.9 mg, 0.08 mmol, 99%).

LC/MS (C ₂₄ H ₂₇ N ₇ O ₂ S ₂ ) 510 [M+H]+; RT 0.857 (LCMS-V-Bi).

¹ H NMR (400 MHz, DMSO) δ 7.91 (d, J=7.9, 1.2 Hz, 1H), 7.68 (d, J=1.2 Hz, 1H), 7.53 (d, J=8.0 Hz, 1H), 7.39 ( ddd, J=8.2, 7.3, 1.3 Hz, 1H), 7.21 (td, J=7.6, 1.2 Hz, 1H), 4.29 (q, J=7.1 Hz, 2H), 3.90-3.67 (m, 4H), 3.18 -3.09 (m, 2H), 2.75-2.63 (m, 2H), 2.46 (d, J=1.0 Hz, 3H), 2.00-1.92 (m, 2H), 1.63 (qd, J=12.4, 3.9 Hz, 2n ), 1.32 (t, J=7.1 Hz, 3H).

Step B: 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl} (methyl)amino)-5 (piperidin-4-yl)-1,3 -thiazole-4-carboxylic acid.

To a solution of the product from step A (40.9 mg, 0.08 mmol, 1 eq.) in 1,4-dioxane (3 ml) was added lithium hydroxide monohydrate (16.8 mg, 0.4 mmol, 5 eq.), and the mixture was refluxed at those

- 158 046336 night. The reaction mixture was allowed to cool to ambient temperature and then concentrated in vacuo. Purification by reverse phase automated flash chromatography (CombiFlash Rf, C18 13 g RediSep column) eluting with a gradient of 5-95% acetonitrile (pH 4; formic acid) in water (pH 4; formic acid) resulted in a solid. which was triturated with chloroform, filtered and dried in vacuo to give the title product as a yellow solid (27.4 mg, 0.06 mmol, 71%).

LC/MS (C ₂₂ H ₂₃ N ₇ O ₂ S ₂ ) 482 [M+H]+; RT 0.772 (LCMS-V-BI).

¹ H NMR (400 MHz, DMSOC) δ 7.83 (d, J=7.7 Hz, 1H), 7.55 (s, 1H), 7.46 (d, J=7.9 Hz, 1H), 7.35-7.27 (m, 1H), 7.16-7.05 (m, 1H), 4.21 (tt, J=12.0, 3.8 Hz, 1H), 3.65 (s, 3H), 3.09-2.95 (m, 2H), 2.62-2.53 (m, 2H), 2.43 ( d, J=1.1 Hz, 3H), 1.83 (dd, J=12.9, 9.4 Hz, 2H), 1.47 (qd, J=12.1, 3.9 Hz, 2H)

HRMS-ERI (m/z) [M+H] ⁺ , calculated for C ₂₂ H ₂₃ N ₇ O ₂ S ₂ : 482.1427; found: 482.1430.

Example 84: 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)-5-{1-[(tert-butoxy)carbonyl]piperidine -4-yl}-1,3-thiazol-4-carboxylic acid

Step A: tert-butyl 4-[2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3yl}(methyl)amino)-4-(ethoxycarbonyl)-1, 3-thiazol-5-yl]piperidine-1-carboxylate.

Tetrabutylammonium fluoride (1 M in tetrahydrofuran, 0.48 ml, 0.48 mmol, 6 eq.) was added to a stirred solution of synthesis 5k, step B (60 mg, 0.08 mmol, 1 eq.) in tetrahydrofuran (3 ml), and the mixture was heated at 55°C overnight. The mixture was allowed to cool to ambient temperature, partitioned between ethyl acetate and saturated aqueous sodium bicarbonate, and the organic phase was dried (magnesium sulfate) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 12 g RediSep™ silica gel cartridge) eluting with a gradient of 0-10% methanol in dichloromethane gave the title product as a yellow glass (37.7 mg, 0.06 mmol, 76%) .

LC/MS (C ₂₉ H ₃₅ N ₇ O ₄ S ₂ ) 610 [M+H] ⁺ ; RT 1.356 (LCMS^-B1).

¹ H NMR (400 MHz, DMSO-de) δ 8.00 (br s, 1H), 7.68 (br s + s, 2H), 7.52-7.351 (m, 1H), 7.31-7.14 (m, 1H), 4.31 ( q, J=7.1 Hz, 2H), 4.22 (d, J=12.8 Hz, 2H), 3.86-3.70 (m, 4H), 2.97-2.71 (m, 2H), 2.46 (s, 3H), 2.051.94 (m, 2H), 1.59-1.46 (m, 2H), 1.44 (s, 9H), 1.35 (t, 3H).

Step B: 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)-5-{1-[(tert-butoxy)carbonyl] piperidine -4-yl}-1,3-thiazol-4-carboxylic acid.

To a solution of the product from step A (47 mg, 0.08 mmol, 1 eq.) in 1,4-dioxane (3 ml) was added lithium hydroxide monohydrate (16.2 mg, 0.39 mmol, 5 eq.), and the mixture was refluxed at during the night. The reaction mixture was allowed to cool to ambient temperature and concentrated in vacuo. Purification by reverse phase automated flash chromatography (CombiFlash Rf, C18 13 g RediSep column) eluting with a gradient of 5-95% acetonitrile (pH 4; formic acid) in water (pH 4; formic acid) provided the desired product as a yellow solid (23 mg, 0.04 mmol, 51%).

HRMS-ERI (m/z) [M+H] ⁺ , calculated for C ₂₇ H ₃₁ N ₇ O ₄ S ₂ : 582.1952; found: 582.1956.

Example 85: 2-[[6-(1,3-Benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]-(3,4dihydroxybutyl)amino]thiazole-4-carboxylic acid

HE

Step A: methyl 2-[tert-butoxycarbonyl-[2-(2,2-dimethyl-1,3-dioxolan-4-yl)ethyl]amino]thiazole 4-carboxylate.

- 159 046336

Using the general Mitsunobu reaction procedure starting with methyl 2-(tert-butoxycarbonylamino)thiazole-4-carboxylate and 2-(2,2-dimethyl-1,3-dioxolan-4-yl)ethanol as starting materials, 3.07 g ( 99%) of the target product.

1H NMR (500 MHz, DMSOC) δ ppm 8.08 (s, 1H), 4.17/4.07 (m+m, 2H), 4.08 (m, 1H), 4.03/3.54 (dd+dd, 2H), 3.80 (s, 3H), 1.88/1.84 (m+m , 2H), 1.53 (s, 9H), 1.28 (s, 3H), 1.22 (s, 3H); MSHR-ERI (m/z): [M+H] ⁺ , calculated for C ₁₇ H ₂₆ N ₂ O ₆ S: 387.1589; found: 387.1585.

Step B: Methyl 2-[2-(2,2-dimethyl-1,3-dioxolan-4-yl)ethylamino]thiazole-4-carboxylate.

Using the general HFIP deprotection procedure, starting with the product from Step A as the appropriate carbamate, 1.21 g (60%) of the desired product was obtained.

1H NMR (500 MHz, DMSOC) δ ppm 7.82 (t, 1H), 7.53 (s, 1H), 4.11 (m, 1H), 4.02/3.49 (dd+dd, 2H), 3.74 (s, 3H), 3.33/3.24 (m+m, 2H), 1.78/1.75 (m+m, 2H), 1.32 (s, 3H), 1.26 (s, 3H); MSHR-ERI (m/z): [M+H] ⁺ , calculated for C ₁₂ H ₁₈ N ₂ O ₄ S: 287.1065; found: 287.1055.

Step C: methyl 2-[(6-chloro-5-methylpyridazin-3-yl)-[2-(2,2-dimethyl-1,3-dioxolan-4yl)ethyl]amino]thiazol-4-carboxylate.

Using the general procedure for the nucleophilic substitution reaction, starting with the product from step B as the starting material, 178 mg (35%) of the desired product was obtained.

1H NMR (500 MHz, DMSOC) δ ppm 8.07 (s, 1H), 7.85 (d, 1H), 4.57/4.37 (m+m, 2H), 4.17 (m, 1H), 4.03/3.62 (dd+dd, 2H), 3.82 (s, 3H), 2.43 (d, 3H), 1.96 (m, 2H), 1.35/1.23 (s+s, 6H); HRMS-ERI (m/z): [M+H] ⁺ , calculated for C ₁₇ H ₂₂ ClN ₄ O ₄ S: 413.1045; found: 413.1048.

Step D: Methyl 2-[[6-(1,3-benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]-[2-(2,2-dimethyl-1,3dioxolan-4-yl)ethyl ]amino]thiazole-4-carboxylate.

Using the general Buchwald reaction procedure II, starting with the product from step C as the starting material and reacting in toluene at 150° C. for 4 hours, 184 mg (81%) of the desired product was obtained.

MSHR-ERI (m/z): [M+H] ⁺ , calculated for C ₂₄ H ₂₇ N ₆ O ₄ S ₂ : 527.1530; found: 527.1512.

Step E: 2-[[6-(1,3-benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]-(3,4dihydroxybutyl)amino]thiazol-4-carboxylic acid.

A 2 M NaOH solution was added to a solution of the product from step D in 1,4-dioxane, and the mixture was stirred at 80°C for 1 hour. After acidifying the mixture with 1N. HCl solution, the reaction mixture was stirred at 80°C for 0.5 h, then the precipitate was filtered and purified using flash column chromatography to obtain the target product.

MSHR-ERI (m/z): [M+H] ⁺ , calculated for C ₂₀ H ₂₁ N ₆ O ₄ S ₂ : 473.1066; found: 473.1055.

Example 86: 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)-5-(1benzylpiperidin-4-yl)-1, 3-thiazole-4-carboxylic acid

Step A: ethyl 5-(1-benzylpiperidin-4-yl)-2-[methyl(5-methyl-6-{[(2Z)-3-{[2(trimethylsilyl)ethoxy]methyl}-2,3- dihydro-1,3-benzothiazol-2-ylidene]amino}pyridazin-3-yl)amino]1,3-thiazol-4-carboxylate.

To a stirred solution of synthesis product 5k (65 mg, 0.1 mmol, 1 eq.) in acetonitrile (5 ml) was added benzaldehyde (0.02 ml, 0.2 mmol, 2 eq.), sodium triacetoxyborohydride (64.6 mg, 0.3 mmol, 3 eq.) and glacial acetic acid (5 μl), and the mixture was stirred at ambient temperature overnight. The reaction mixture was partitioned between ethyl acetate and saturated aqueous sodium bicarbonate, and the organic phase was washed with brine, dried (magnesium sulfate) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 12 g RediSep™ silica gel cartridge) eluting with a gradient of 0-60% ethyl acetate in isoheptane gave the title product as a colorless gum (44.7 mg, 0.06 mmol, 60%).

LC/MS (C ₃₇ H ₄₇ N ₇ O ₃ SiS ₂ ) 730 [M+H] ⁺ ; RT 1.32 (LCMS-V-BI).

1H NMR (400 MHz, DMSO) δ 7.89 (d, 1H), 7.66 (d, J=1.1 Hz, 1H), 7.51-7.41 (m, 2H), 7.40-7.31 (m, 4H), 7.30-7.22 ( m, 2H), 5.86 (s, 2H), 4.29 (q, J=7.1 Hz, 2H), 3.76 (s, 3H), 3.76-3.69 (m, 2H), 3.63 (tt, J=12.3, 3.8 Hz , 1H), 3.51 (s, 2H), 2.94 (d, J=11.1 Hz, 2H), 2.46 (d, J=1.0 Hz, 3H), 2.15-1.89 (m, 4H), 1.67 (qd, J= 12.2, 3.6 Hz, 2H), 1.32 (t, J=7.1 Hz, 3H), 0.92 (dd, J=8.6, 7.4 Hz, 2H), -0.11 (s, 9H).

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Step B: ethyl 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)-5-(1benzylpiperidin-4-yl)-1 ,3-thiazole-4-carboxylate.

Hydrochloric acid (4 M in 1,4-dioxane; 3 mL, 12 mmol, 196 eq.) was added to a stirred solution of the product from step A (44.7 mg, 0.06 mmol, 1 eq.) in 1,4-dioxane (3 mL ), and the mixture was stirred at 60°C overnight. The reaction mixture was allowed to cool to ambient temperature, partitioned between dichloromethane and saturated aqueous sodium bicarbonate, and the organic phase was washed with brine, dried (magnesium sulfate) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 12 g RediSep™ silica gel cartridge) eluting with a gradient of 0-5% methanol in dichloromethane gave the title product as a yellow gum (22 mg, 0.04 mmol, 60%).

LC/MS (C ₃₁ H ₃ 3N ₇ O ₂ S ₂ ) 600 [M+H]+; RT 0.99 (LCMS-V-BI).

1H NMR (400 MHz, DMSO-de) δ 11.05 (br s, 1H), 7.98 (br s, 1H), 7.68 (br s + s, 2H), 7.43-7.31 (m, 5H), 7.30-7.19 ( m, 2H), 4.29 (q, J=7.1 Hz, 2H), 3.76 (s, 3H), 3.63 (tt, J=12.0, 3.8 Hz, 1H), 3.52 (s, 2H), 2.98-2.91 (m , 2H), 2.46 (s, 3H), 2.05 (t, 2H), 2.01-1.94 (m, 2H), 1.67 (qd, J=12.2, 3.6 Hz, 2H), 1.32 (t, J=7.1 Hz, 3H).

Step C: 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl} (methyl)amino)-5 -(1benzylpiperidin-4-yl)-1, 3-thiazole-4-carboxylic acid.

To a solution of the product from step B (22 mg, 0.04 mmol, 1 eq.) in 1,4-dioxane (3 ml) was added lithium hydroxide monohydrate (7.7 mg, 0.18 mmol, 5 eq.), and the mixture was refluxed at overnight, then allowed to cool to ambient temperature and concentrated in vacuo. The resulting solid was triturated with water, then with diethyl ether, filtered and dried in vacuo to give the title product as a yellow solid (9.6 mg, 0.02 mmol, 46%).

HRMS-ERI (m/z) [M+H]+, calculated for C ₂₉ H ₃₀ N ₇ O ₂ S ₂ : 572.1897; found: 572.1903.

Example 87: 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)-5-[1-(2methylpropyl)piperidin-4- yl]-1,3-thiazol-4-carboxylic acid

Step A: ethyl 2-[methyl(5-methyl-6-{[(2Z)-3-{[2-(trimethylsilyl)ethoxy]methyl}-2,3-dihydro-1,3benzothiazol-2-ylidene]amino }pyridazin-3-yl)amino]-5-[1-(2-methylpropyl)piperidin-4-yl]-1,3-thiazol4-carboxylate.

Isobutyraldehyde (0.04 ml, 0.42 mmol, 2 equiv.), sodium triacetoxyborohydride (133 mg, 0.63 mmol, 3 eq.) and glacial acetic acid (10 μl), and the mixture was stirred at ambient temperature overnight. The reaction mixture was partitioned between dichloromethane and saturated aqueous sodium bicarbonate, and the organic phase was washed with brine, dried (magnesium sulfate) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 12 g RediSep™ silica gel cartridge) eluting with a gradient of 0-60% ethyl acetate in isoheptane gave the title product as an off-white solid (114 mg, 0.16 mmol, 79 %).

LC/MS (C ₃ 4H4 ₉ N ₇ O ₃ SiS ₂ ) 696 [M+H]+; RT 1.30 (LCMS-V-BI).

1H NMR (400 MHz, DMSO-d^ δ 7.92-7.85 (m, 1H), 7.66 (d, J=1.1 Hz, 1H), 7.50-7.38 (m, 2H), 7.29-7.20 (m, 1H), 5.86 (s, 2H), 4.29 (q, J=7.1 Hz, 2H), 3.76 (s, 3H), 3.75-3.68 (m, 2H), 3.66-3.53 (m, 1H), 2.99-2.90 (m, 2H), 2.46 (d, J=1.0 Hz, 3H), 2.07 (d, J=7.4 Hz, 2H), 2.00-1.88 (m, 4H), 1.79 (hept, J=6.7 Hz, 1H), 1.73- 1.59 (m, 2H), 1.32 (t, J=7.1 Hz, 3H), 0.97-0.85 (m, 8H), -0.11 (s, 9H).

Step B: ethyl 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)-5-[1-(2methylpropyl)piperidin-4 -yl]-1,3-thiazol-4-carboxylate.

Trifluoroacetic acid (1.2 mL, 16.1 mmol, 98 eq.) was added to a stirred solution of the product from Step A (114 mg, 0.16 mmol, 1 eq.) in dichloromethane (4 mL), and the mixture was stirred at ambient temperature overnight. The reaction mixture was allowed to cool to ambient temperature, then partitioned between dichloromethane and saturated aqueous sodium bicarbonate, and the organic phase was washed with brine, dried (magnesium sulfate) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 12 g RediSep™ silica gel cartridge) eluting with a gradient of 0-5% methanol in dichloromethane provided the title product as a yellow solid (76.8 mg, 0.14 mmol, 83%) .

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LC/MS (C ₂₈ H ₃₅ N ₇ O ₂ S ₂ ) 566 [M+H]+; RT 0.96 (LCMS-ν-Βη.

1H NMR (400 MHz, DMSO-de) δ 7.96 (br s, 1H), 7.68 (s, 1H), 7.56 (br s, 1H), 7.39 (t, J=7.6 Hz, 1H), 7.21 (t, J=7.4 Hz, 1H), 4.30 (q, J=7.1 Hz, 2H), 3.76 (s, 3H), 3.67-3.55 (m, 1H), 2.96 (d, J=11.5 Hz, 2H), 2.47 ( s, 3H), 2.08 (d, J=7.4 Hz, 2H), 2.02-1.89 (m, 4H), 1.80 (hept, J=6.7 Hz, 1H), 1.66 (td, J=14.3, 13.7, 10.3 Hz , 2H), 1.33 (t, J=7.1 Hz, 3H), 0.90 (d, J=6.5 Hz, 6H).

Step C: 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)-5-[1-(2methylpropyl)piperidin-4- yl]-1,3-thiazol-4-carboxylic acid.

To a solution of the product from step B (76.8 mg, 0.14 mmol, 1 eq.) in 1,4-dioxane (5 ml) was added lithium hydroxide monohydrate (28.5 mg, 0.68 mmol, 5 eq.) and the mixture was refluxed at during the night. The reaction mixture was allowed to cool to ambient temperature and concentrated in vacuo. Purification by reverse phase automated flash chromatography (CombiFlash Rf, C18 13 g RediSep column) eluting with a gradient of 5-95% acetonitrile in water gave the title product as a yellow solid (50 mg, 0.09 mmol, 69% ).

HRMS-ERI (m/z) [M+H] ⁺ , calculated for C ₂₆ H ₃₂ N ₇ O ₂ S ₂ : 538.2053; found: 538.2060.

Example 88: 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)-5-[4(phenoxymethyl)piperidin-1 -yl ]-1,3-thiazole-4-carboxylic acid

Step A: ethyl 2-[methyl(5-methyl-6-{[(2Z)-3-{[2-(trimethylsilyl)ethoxy]methyl}-2,3-dihydro-1,3benzothiazol-2-ylidene]amino }pyridazin-3-yl)amino]-5-[4-(phenoxymethyl)piperidin-1-yl]-1,3-thiazol-4carboxylate.

To a stirred solution of synthesis product 11b (200 mg, 0.31 mmol, 1 eq.), 4(phenoxymethyl)piperidine hydrochloride (143 mg, 0.63 mmol, 2 eq.) and cesium carbonate (308 mg, 0.94 mmol, 3 eq.) in rac-BINAP-Pd-G3 (31.2 mg, 0.03 mmol, 0.1 equiv) was added to 1,4-dioxane (8 ml) under nitrogen atmosphere and the mixture was heated at 100°C overnight. The reaction mixture was allowed to cool to ambient temperature, then diluted with ethyl acetate, washed with water, then brine, dried (magnesium sulfate) and concentrated in vacuo to give the title product as a yellow foam (153 mg, 0.2 mmol, 65%).

LC/MS (C ₃₇ H ₄₇ N ₇ O ₄ SiS ₂ ) 746 [M+H] ⁺ ; RT 1.60 (ZHMS-U-Y).

1H NMR (400 MHz, DMSO^) δ 7.82 (d, 1H), 7.61 (d, J=1.1 Hz, 1H), 7.50-7.39 (m, 2H), 7.33-7.20 (m, 3H), 6.99-6.90 (m, 3H), 5.85 (s, 2H), 4.25 (q, J=7.1 Hz, 2H), 3.91 (d, J=6.1 Hz, 2H), 3.79-3.68 (m, 5H), 3.55 (d, J=11.2 Hz, 2H), 2.88 (td, J=11.6, 2.3 Hz, 2H), 2.44 (d, J=0.9 Hz, 3H), 2.00-1.82 (m, 3H), 1.54 (qd, J=12.0 , 3.9 Hz, 2H), 1.31 (t, J=7.1 Hz, 3H), 0.95-0.89 (m, 2H), -0.11 (s, 9H).

Step B: ethyl 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)-5-[4(phenoxymethyl)piperidin-1- yl]-1,3-thiazol-4-carboxylate.

Hydrochloric acid (4 M in 1,4-dioxane; 3 mL, 12 mmol, 58.7 eq.) was added to a stirred solution of the product from step A (153 mg, 0.2 mmol, 1 eq.) in dichloromethane (10 mL), and the mixture stirred at 50°C overnight. The reaction mixture was allowed to cool to ambient temperature, then partitioned between dichloromethane and saturated aqueous sodium bicarbonate, and the organic phase was washed with brine, dried (magnesium sulfate) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 12 g RediSep™ silica gel cartridge) eluting with a gradient of 0-5% methanol in dichloromethane gave the title product as a yellow glass (83.8 mg, 0.14 mmol, 67%) .

LC/MS (C ₃₁ H ₃₃ N ₇ O ₃ S ₂ ) 616 [M+H] ⁺ ; RT 1.39 (LCMS-V-BI).

1H NMR (400 MHz, DMSO^) δ 11.01 (br s, 1H), 7.94 (br s, 1H), 7.63 (br s + s, 2H), 7.42-7.35 (m, 1H), 7.34-7.26 (m , 2H), 7.23-7.15 (m, 1H), 7.00-6.88 (m, 3H), 4.25 (q, J=7.1 Hz, 2H), 3.91 (d, J=6.1 Hz, 2H), 3.70 (s, 3H), 3.55 (d, J=14.2 Hz, 2H), 2.88 (dd, J=12.2, 9.8 Hz, 2H), 2.45 (s, 3H), 2.02-1.81 (m, 3H), 1.62-1.47 (m , 2H), 1.30 (t, J=7.1 Hz, 3H).

Step C: 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)-5-[4(phenoxymethyl)piperidin-1-yl ]-1,3-thiazole-4-carboxylic acid.

To a solution of the product from step B (83.8 mg, 0.14 mmol, 1 eq.) in 1,4-dioxane (3 ml) was added lithium hydroxide monohydrate (28.6 mg, 0.68 mmol, 5 eq.) and the mixture was refluxed at during the night. The reaction mixture was allowed to cool to ambient temperature and concentration

- 162 046336 dug in a vacuum. Purification by reverse phase automated flash chromatography (CombiFlash Rf, C18 13 g RediSep column) eluting with a gradient of 5-95% acetonitrile in water gave the title product as a yellow solid (59.7 mg, 0.1 mmol, 75% ).

HRMS-ERI (m/z) [M+H]+, calculated for C ₂₉ H ₃₀ N ₇ O ₃ S2: 588.1846; found: 588.1853.

Example 89: 5-(1-benzoylpiperidin-4-yl)-2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3yl} (methyl)amino)-1, 3-thiazole-4-carboxylic acid

Step A: ethyl 5-(1-benzoylpiperidin-4-yl)-2-[methyl(5-methyl-6-{[(27)-3-{[2(trimethylsilyl)ethoxy]methyl}-2,3- dihydro-1,3-benzothiazol-2-ylidene]amino}pyridazin-3-yl)amino]1,3-thiazol-4-carboxylate.

To a stirred solution of synthesis product 5k (88 mg, 0.14 mmol, 1 eq.) and triethylamine (0.04 ml, 0.28 mmol, 2 eq.) in dichloromethane (3 ml) was added benzoyl chloride (0.02 ml, 0.17 mmol, 1.2 eq.), and the mixture was stirred at ambient temperature for 4.5 hours and then concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 12 g RediSep™ silica gel cartridge) eluting with a gradient of 0-100% ethyl acetate in isoheptane gave the title product as an off-white foam (89.6 mg, 0.12 mmol, 88% ).

LC/MS (C ₃₇ H ₄₅ N ₇ O ₄ SiS ₂ ) 744 [M+H]+; RT 1.49 (LCMS-V-BI).

1H NMR (400 MHz, DMSO-dg) δ 7.88 (d, J=7.7 Hz, 1H), 7.67 (d, J=1.2 Hz, 1H), 7.51-7.40 (m, 7H), 7.25 (ddd, J= 8.1, 7.0, 1.5 Hz, 1H), 5.86 (s, 2H), 4.66 (br s, 1H), 4.30 (q, J=7.1 Hz, 2H), 3.97-3.85 (m, 1H), 3.77 (s, 3H), 3.75-3.67 (m, 2H), 3.19 (br s, 1H), 2.88 (br s, 1H), 2.47 (d, J=0.9 Hz, 3H), 2.10-1.87 (m, 2H), 1.77 -1.55 (m, 2H), 1.35 (t, J=7.1 Hz, 3H), 0.98-0.88 (m, 2H), -0.07 (s, 9H).

Step B: ethyl 5-(1-benzoylpiperidin-4-yl)-2-({6-[(1,3-benzothiazol-2-yl)amino]-5methylpyridazin-3-yl} (methyl)amino)-1 ,3-thiazole-4-carboxylate.

Trifluoroacetic acid (1.2 mL, 16.1 mmol, 134 eq.) was added to a stirred solution of the product from Step A (89.6 mg, 0.12 mmol, 1 eq.) in dichloromethane (4 mL), and the mixture was stirred at ambient temperature overnight. The reaction mixture was partitioned between dichloromethane and saturated aqueous sodium bicarbonate, and the organic phase was washed with brine, dried (magnesium sulfate) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 12 g RediSep™ silica gel cartridge) eluting with a gradient of 0-5% methanol in dichloromethane gave the title product as a yellow glass (55 mg, 0.09 mmol, 74%) .

LC/MS (C ₃₁ H ₃₁ N ₇ O ₃ S ₂ ) 614 [M+H]+; RT 1.41 (LCMS-V-BI).

1H NMR (400 MHz, DMSO-dg) δ 11.08 (br s, 1H), 7.96 (br s, 1H), 7.69 (br s + s, 2H), 7.64-7.43 (m, 5H), 7.40 (t, J=7.5 Hz, 1H), 7.23 (t, 1H), 4.69 (br s, 1H), 4.31 (q, J=7.1 Hz, 2H), 4.00-3.80 (m, 1H), 3.82-3.66 (m, 4H), 3.30-2.74 (m, 2H), 2.47 (s, 3H), 2.21-1.84 (m, 2H), 1.76-1.57 (m, 2H), 1.33 (t, J=7.1 Hz, 3H).

Step C: 5-(1-benzoylpiperidin-4-yl)-2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3yl}(methyl)amino)-1, 3-thiazole-4-carboxylic acid.

To a solution of the product from step B (55 mg, 0.09 mmol, 1 eq.) in 1,4-dioxane (4 ml) was added lithium hydroxide monohydrate (18.8 mg, 0.45 mmol, 5 eq.), and the mixture was refluxed at during the night. The reaction mixture was allowed to cool to ambient temperature and concentrated in vacuo. Purification by reverse phase automated flash chromatography (CombiFlash Rf, C18 13 g RediSep column) eluting with a gradient of 5-95% acetonitrile in water gave the title product as a yellow solid (35.8 mg, 0.06 mmol, 68% ).

HRMS-ERI (m/z) [M+H]+, calculated for C ₂₉ H ₂₈ N ₇ O ₃ S ₂ : 586.1690; found: 586.1702.

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Example 90: 3-{1-[(adamantan-1-yl)methyl]-5-methyl-1H-pyrazol-4-yl}-6-({6-[(1,3-benzothiazol-2yl)amino] -5-methylpyridazin-3-yl}(methyl)amino)pyridine-2-carboxylic acid

Step A: ethyl 3-bromo-6-{[(tert-butoxy)carbonyl](methyl)amino}pyridine-2-carboxylate.

Cesium carbonate (4.9 g, 15.1 mmol, 1.5 eq.) was added under nitrogen atmosphere to a solution of ethyl 3,6dibromopyridine-2-carboxylate (3.1 g, 10.03 mmol, 1 eq.), tert-butyl N-methylcarbamate (1.65 g, 12.6 mmol, 1.25 eq.), tris(dibenzylideneacetone)dipalladium(0) (91.9 mg, 0.1 mmol, 0.01 eq.) and xantphos (116 mg, 0.2 mmol, 0.02 eq.) in 1,4-dioxane (40 ml), and the mixture was heated at 100°C overnight. The reaction mixture was allowed to cool to ambient temperature and ethyl acetate (250 ml) was added. The mixture was washed with water (2x100 ml), brine (100 ml), dried (magnesium sulfate) and concentrated in vacuo. Purification by flash column chromatography (100 g silica gel) eluting with dichloromethane gave the title product as a pale green oil (1.78 g, 4.96 mmol, 49%).

LC/MS (C ₁₄ H ₁₉ BrN ₂ O ₄ ) 303 [M-tBu+H]+; RT 1.43 (LCMS-V-BI).

¹ H NMR (400 MHz, CDCl ₃ ) δ 7.85-7.74 (m, 2H), 4.45 (q, J=7.1 Hz, 2H), 3.40 (s, 3H), 1.52 (s, 9H), 1.42 (t, J=7.1 Hz, 3H).

Step B: ethyl 3-bromo-6-(methylamino)pyridine-2-carboxylate.

Trifluoroacetic acid (1.9 mL, 24.8 mmol, 5 eq.) was added at 0°C to a solution of the product from step A (1.78 g, 4.96 mmol, 1 eq.) in dichloromethane (15 mL) and the mixture was allowed to warm to ambient temperature and stirred overnight. The reaction mixture was partitioned between dichloromethane (50 ml) and saturated aqueous sodium bicarbonate and the organic phase was washed with water (2x50 ml) and brine (50 ml), dried (magnesium sulfate) and concentrated in vacuo to give the title product as a yellow oil (1.2 g, 4.63 mmol, 94%).

LC/MS (C ₉ H ₁₁ BrN ₂ O ₂ ) 260 [M+H]+; RT 1.11 (LCMS-V-BI).

1H NMR (400 MHz, CDCl ₃ ) δ 7.61 (d, J=8.9 Hz, 1H), 6.36 (d, J=8.9 Hz, 1H), 4.82 (s, 1H), 4.43 (q, J=7.1 Hz, 2H), 2.90 (d, J=4.3 Hz, 3H), 1.41 (t, J=7.1 Hz, 3H).

Step C: ethyl 3-bromo-6-[(6-chloro-1,2,4,5-tetrazin-3-yl) (methyl) amino]pyridine-2-carboxylate.

A solution of 3,6-dichloro-1,2,4,5-tetrazine (664 mg, 4.4 mmol, 0.95 eq.) and the product from step B (1.2 g, 4.63 mmol, 1 eq.) in acetonitrile (20 ml) was heated in a stoppered flask for 18 hours. The reaction mixture was allowed to cool to ambient temperature and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 40 g RediSep™ silica gel cartridge) eluting with a gradient of 0-15% ethyl acetate in isoheptane gave the title product as a red solid (898 mg, 2.4 mmol, 55%) .

LC/MS (C ₁₁ H ₁₀ BrClN ₆ O ₂ ) 373 [M+H]+; RT 1.31 (LCMS-V-BI).

1H NMR (400 MHz, CDCl ₃ ) δ 7.99 (d, J=8.7 Hz, 1H), 7.69 (d, J=8.7 Hz, 1H), 4.49 (q, J=7.1 Hz, 2H), 3.87 (s, 3H), 1.44 (t, J=7.1 Hz, 3H).

Step D: Ethyl 3-bromo-6-[(6-chloro-5-methylpyridazin-3-yl) (methyl) amino]pyridine-2-carboxylate.

Ethyl 1-propenyl ether (2.64 mL, 23.8 mmol, 10 eq.) was added to a solution of the product from Step C (890 mg, 2.38 mmol, 1 eq.) in 1,4-dioxane (15 mL) and the mixture was heated to sealed tube at 100°C for 24 hours. The reaction mixture was allowed to cool to ambient temperature and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 80 g RediSep™ silica gel cartridge) eluting with a gradient of 0-50% ethyl acetate in isoheptane provided the title product as an orange solid (776 mg, 2.01 mmol, 85%) .

LC/MS (C ₁₄ H ₁₄ BrClN ₄ O ₂ ) 385 [M+H]+; RT 1.29 (LCMS-V-BI).

1H NMR (400 MHz, CDCl ₃ ) δ 7.86 (d, J=8.9 Hz, 1H), 7.65 (q, J=1.0 Hz, 1H), 7.15 (d, J=8.8 Hz, 1H), 4.44 (q, J=7.1 Hz, 2H), 3.70 (s, 3H), 2.35 (d, J=1.0 Hz, 3H), 1.42 (t, J=7.1 Hz, 3H).

Step E: ethyl 3-{1-[(adamantan-1-yl)methyl]-5-methyl-1H-pyrazol-4-yl}-6-[(6-chloro-5methylpyridazin-3-yl)(methyl) amino]pyridine-2-carboxylate.

A solution of potassium carbonate (552 mg, 3.99 mmol, 2 eq.) in water (2 ml), and the mixture was bubbled with nitrogen (10 min). Pd(dppf)Cl ₂ .CH ₂ Cl ₂ (163 mg, 0.2 mmol, 0.1 eq.) was added and the mixture was refluxed overnight. The reaction mixture was allowed to cool to ambient temperature, then partitioned between ethyl acetate and hydrochloric acid.

- 164 046336 solution, and the organic phase was dried (magnesium sulfate) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 40 g RediSep™ silica gel cartridge) eluting with a gradient of 0-60% ethyl acetate in isoheptane provided the title product as a cream solid (680 mg, 1.27 mmol, 64%) .

LC/MS (C ₂₉ H ₃₅ ClN ₆ O ₂ ) 535 [M+H]+; RT 1.55 (LCMS-V-BI).

¹ H NMR (400 MHz, DMSO^6) δ 7.82-7.73 (m, 2H), 7.50 (d, J=8.6 Hz, 1H), 7.35 (s, 1H), 4.14 (q, J=7.1 Hz, 2H ), 3.77 (s, 2H), 3.63 (s, 3H), 2.32 (d, J=1.0 Hz, 3H), 2.18 (s, 3H), 2.01-1.88 (m, 3H), 1.72-1.62 (m, 3H), 1.61-1.52 (m, 9H), 1.13 (t, J=7.1 Hz, 3H).

Step F: ethyl 3-{1-[(adamantan-1-yl)methyl]-5-methyl-1H-pyrazol-4-yl}-6-({6-[(1,3-benzothiazol-2yl)amino ]-5-methylpyridazin-3-yl}(methyl)amino)pyridine-2-carboxylate.

N,N-diisopropylethylamine (0.59 ml, 3.36 mmol, 3 eq.), xanthos (64.9 mg, 0.11 mmol, 0.1 eq.) and tris(dibenzylideneacetone)dipalladium(0) (51.3 mg, 0.06 mmol, 0.05 eq.), and the mixture was heated in a sealed tube at for 64 hours. The reaction mixture was allowed to cool to ambient temperature, then diluted with ethyl acetate, washed with brine and concentrated in vacuo. Purification by reverse phase automated flash chromatography (CombiFlash Rf, C18 130 g RediSep column) eluting with a gradient of 30-95% acetonitrile in water gave the title product as a yellow glassy solid (513 mg, 0.79 mmol, 71%).

LC/MS (C ₃₆ H ₄₀ N ₈ O ₂ S) 649 [M+H]+; RT 1.62 (Zh'HMS-\'-B1).

^1H NMR (400 MHz, DMSO- ₀₆ ) δ 11.09 (br s, 1H), 7.89 (br s, 1H), 7.74-7.51 (m, 3H), 7.40 (t, 1H), 7.33 (s, 1H ), 7.30 (d, 1H), 7.22 (t, 1H), 4.13 (q, J=7.1 Hz, 2H), 3.76 (s, 2H), 3.61 (s, 3H), 2.39 (s, 3H), 2.17 (s, 3H), 1.95 (s, 3H), 1.74-1.62 (m, 3H), 1.62-1.46 (m, 9H), 1.12 (t, J=7.1 Hz, 3H).

Step G: 3-{1-[(adamantan-1-yl)methyl]-5-methyl-1H-pyrazol-4-yl}-6-({6-[(1,3-benzothiazol-2yl)amino] -5-methylpyridazin-3-yl}(methyl)amino)pyridine-2-carboxylic acid.

To a solution of the product from step F (510 mg, 0.79 mmol, 1 eq.) in 1,4-dioxane (10 ml) was added lithium hydroxide monohydrate (82.5 mg, 1.97 mmol, 2.5 eq.) and the mixture was refluxed at for 1 hour. The reaction mixture was allowed to cool to ambient temperature and concentrated in vacuo, then water (5 ml) was added and the pH was adjusted to 3 by adding 2 M aqueous hydrochloric acid. The mixture was extracted into dichloromethane and the organic extract was washed with brine, dried (magnesium sulfate) and concentrated in vacuo. The resulting solid was triturated with methanol, filtered, washed with methanol and dried in vacuo to give the title product as an off-white solid (362 mg, 0.58 mmol, 74%).

HRMS-ERI (m/z) [M+H]+, calculated for C ₃₄ H ₃₇ N ₈ O ₂ S: 621.2755; found: 621.2765.

Example 91: 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)-5-(1phenylpiperidin-4-yl)-1, 3-thiazole-4-carboxylic acid

Step A: ethyl 2-[methyl(5-methyl-6-{[(2Z)-3-{[2-(trimethylsilyl)ethoxy]methyl}-2,3-dihydro-1,3benzothiazol-2-ylidene]amino } pyridazin-3 -yl)amino]-5-(1-phenylpiperidin-4-yl)-1,3-thiazol-4carboxylate.

Tris(dibenzylideneacetone)dipalladium(0) (10.7 mg, 0.01 mmol, 0.05 eq.) under a nitrogen atmosphere was added to a stirred solution of the synthesis product 5k (150 mg, 0.23 mmol, 1 eq.), bromobenzene (0.04 ml, 0.35 mmol, 1.5 eq.), cesium carbonate (229 mg, 0.7 mmol, 3 eq.) and xanthos (13.6 mg, 0.02 mmol, 0.1 eq.) in 1,4-dioxane (5 ml), and the mixture was heated at 100°C for nights. The reaction mixture was allowed to cool to ambient temperature, then partitioned between dichloromethane and water, and the organic phase was washed with water, then brine, dried (magnesium sulfate) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 12 g RediSep™ silica gel cartridge) eluting with a gradient of 0-50% ethyl acetate in isoheptane gave the title product as a brown foam (156 mg, 0.22 mmol, 93%).

LC/MS (C ₃₆ H ₄₅ N ₇ O ₃ SiS ₂ ) 716 [M+H]+; RT 1.68 (LCMS-V-BI).

^1H NMR (400 MHz, DMSO^) δ 7.84 (d, 1H), 7.67 (d, J=1.1 Hz, 1H), 7.50-7.39 (m, 2H), 7.30-7.18 (m, 3H), 7.04- 6.96 (m, 2H), 6.79 (t, 1H), 5.86 (s, 2H), 4.31 (q, J=7.1 Hz, 2H), 3.94-3.64 (m, 8H), 2.83-2.68 (m, 2H) , 2.46 (d, J=1.0 Hz, 3H), 2.10 (d, J=12.1 Hz, 2H), 1.88-1.71 (m, 2H), 1.34 (t, J=7.1 Hz, 3H), 1.00-0.86 ( m, 2H), -

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0.11 (s, 9H).

Step B: ethyl 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)-5-(1phenylpiperidin-4-yl)-1 ,3-thiazole-4-carboxylate.

Trifluoroacetic acid (1.2 mL, 16.1 mmol, 74 eq.) was added to a stirred solution of the product from Step A (156 mg, 0.22 mmol, 1 eq.) in dichloromethane (6 mL), and the mixture was stirred at ambient temperature overnight. The reaction mixture was partitioned between dichloromethane and saturated aqueous sodium bicarbonate, and the organic phase was washed with brine, dried (magnesium sulfate) and concentrated in vacuo. The resulting solid was triturated with 1:1 dichloromethane/heptane, filtered and dried in vacuo to give the title product as a yellow solid (92 mg, 0.16 mmol, 72%).

LC/MS (C ₃₀ H ₃₁ N ₇ O ₂ S ₂ ) 586 [M+H]+; RT 1.40 (LCMS-V-BI).

1H NMR (400 MHz, DMSO-de) δ 12.44 (br s, 1H), 11.02 (br s, 1H), 7.98 (br s, 1H), 7.81-7.54 (m, 2H), 7.38 (br s, 1H ), 7.30-7.12 (m, 3H), 7.05-6.97 (m, 2H), 6.80 (t, 1H), 4.31 (q, J=7.1 Hz, 2H), 3.91-3.69 (m, 6H), 2.82- 2.70 (m, 2H), 2.47 (s, 3H), 2.11 (d, J=12.4 Hz, 2H), 1.88-1.69 (m, 2H), 1.33 (t, J=7.1 Hz, 3H).

Step C: 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)-5-(1phenylpiperidin-4-yl)-1, 3-thiazole-4-carboxylic acid.

To a solution of the product from step B (92 mg, 0.16 mmol, 1 eq.) in 1,4-dioxane (3 ml) was added lithium hydroxide monohydrate (33 mg, 0.79 mmol, 5 eq.) and the mixture was refluxed at during the night. The reaction mixture was allowed to cool to ambient temperature and the solids were collected by filtration, washed with 1,4-dioxane, then triturated with water, filtered and dried in vacuo to give the title product as a yellow solid (50.3 mg, 0.09 mmol, 57 %) [as lithium salt].

HRMS-ERI (m/z) [M+H] ⁺ , calculated for C ₂₈ H ₂₈ N ₇ O ₂ S ₂ : 558.1740; found: 558.1749.

Example 92: 5-[1-(Benzenesulfonyl)piperidin-4-yl]-2-({6-[(1,3-benzothiazol-2-yl)amino]-5methylpyridazin-3-yl}(methyl)amino) -1,3-thiazole-4-carboxylic acid

Step A: ethyl 5-[1-(benzenesulfonyl)piperidin-4-yl]-2-[methyl(5-methyl-6-{[(2Z)-3-{[2(trimethylsilyl)ethoxy]methyl }—2 ,3-dihydro-1,3-benzothiazol-2-ylidene] amino }pyridazin-3-yl)amino] 1,3-thiazol-4-carboxylate.

To a stirred solution of synthesis product 5k (200 mg, 0.31 mmol, 1 eq.) and triethylamine (0.09 ml, 0.63 mmol, 2 eq.) in dichloromethane (5 ml) was added benzenesulfonyl chloride (0.05 ml, 0.38 mmol, 1.2 eq.) and a catalytic amount of 4-(dimethylamino)pyridine, and the mixture was stirred at ambient temperature overnight. The reaction mixture was partitioned between dichloromethane and water, and the organic phase was washed with brine, dried (magnesium sulfate) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 12 g RediSep™ silica gel cartridge) eluting with a gradient of 0-60% ethyl acetate in isoheptane provided the title product as a cream solid (204 mg, 0.26 mmol, 84%) .

LC/MS (C ₃₆ H ₄₅ N ₇ O ₅ SiS ₃ ) 780 [M+H] ⁺ ; RT 1.53 (LCMS-V-BI).

1H NMR (400 MHz, DMSO-dg) δ 7.94-7.87 (m, 1H), 7.85-7.64 (m, 6H), 7.51-7.40 (m, 2H), 7.31-7.22 (m, 1H), 5.87 (s , 2H), 4.24 (q, J=7.1 Hz, 2H), 3.84 (d, J=11.8 Hz, 2H), 3.78-3.68 (m, 5H), 3.61-3.49 (m, 1H), 2.46 (d, J=0.9 Hz, 3H), 2.39-2.29 (m, 2H), 2.05 (d, J=11.2 Hz, 2H), 1.77-1.61 (m, 2H), 1.25 (t, J=7.1 Hz, 3H), 0.97-0.88 (m, 2H), -0.11 (s, 9H).

Step B: ethyl 5-[1-(benzenesulfonyl)piperidin-4-yl]-2-({6-[(1,3-benzothiazol-2-yl)amino]-5methylpyridazin-3-yl}(methyl)amino )-1,3-thiazole-4-carboxylate.

Trifluoroacetic acid (1 mL, 13.4 mmol, 51 eq.) was added to a stirred solution of the product from Step A (204 mg, 0.26 mmol, 1 eq.) in dichloromethane (6 mL) and the mixture was stirred at ambient temperature overnight. The reaction mixture was partitioned between dichloromethane and saturated aqueous sodium bicarbonate, and the organic phase was washed with brine, dried (magnesium sulfate) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 12 g RediSep™ silica gel cartridge) eluting with a gradient of 0-5% methanol in dichloromethane gave the title product as a yellow glass (121 mg, 0.19 mmol, 71%) .

LC/MS (C ₃₀ H ₃₁ N ₇ O ₄ S ₃ ) 650 [M+H] ⁺ ; RT 1.28 (LCMS-V-BI).

1H NMR (400 MHz, DMSO-do) δ 12.44 (br s, 1H), 11.06 (br s, 1H), 8.14-7.86 (m, 2H), 7.86-7.63 (m, 6H), 7.45-7.34 (m , 1H), 7.28-7.16 (m, 1H), 4.23 (q, J=7.1 Hz, 2H), 3.88-3.80 (m, 2H), 3.75 (s, 3H), 3.62-3.47 (m,

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1H), 2.46 (s, 3H), 2.40-2.29 (m, 2H), 2.04 (d, J=12.5 Hz, 2H), 1.75-1.58 (m, 2H), 1.25 (t, J=7.1 Hz, 3H ).

Step C: 5-[1 -(benzenesulfonyl)piperidin-4-yl]-2-({6-[(1,3-benzothiazol-2-yl)amino]-5methylpyridazin-3-yl} (methyl)amino) -1,3-thiazole-4-carboxylic acid.

To a solution of the product from step B (121 mg, 0.19 mmol, 1 eq.) in 1,4-dioxane (3 ml) was added lithium hydroxide monohydrate (39.1 mg, 0.93 mmol, 5 eq.), and the mixture was refluxed at during the night. The reaction mixture was allowed to cool to ambient temperature and the solids were collected by filtration and washed with 1,4-dioxane. The material was triturated with water, filtered and dried in vacuo to give the title product as a yellow solid (102 mg, 0.16 mmol, 88%) [as lithium salt].

HRMS-ERI (m/z) [M+H]+, calculated for C ₂₈ H ₂₈ N ₇ O ₄ S ₃ : 622.1359; found: 622.1367.

Example 93: 2-[[6-(1,3-Benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]-(3-hydroxy-4methoxybutyl)amino]thiazol-4-carboxylic acid

Step A: Methyl 2-[tert-butoxycarbonyl-[3-[tert-butyl(dimethyl)silyl]oxy-4methoxybutyl]amino]thiazole-4-carboxylate.

Using the general Mitsunobu reaction procedure, starting with methyl 2-(tert-butoxycarbonylamino)thiazole-4-carboxylate and synthesis product 2h as starting materials and carrying out the reaction in THF at room temperature. within 1 hour, 4.16 g (81%) of the target product was obtained.

¹ H NMR (500 MHz, DMSO^) δ ppm 8.08 (s, 1H), 4.14/4.07 (m+m, 2H), 3.86 (m, 1H), 3.79 (s, 3H), 3.35/3.29 (dd+dd, 2H), 3.26 (s, 3H), 1.83/1.74 (m+m, 2H), 1.53 (s, 9H), 0.85 (s, 9H), 0.04/0.03 (s+s, 6H). ¹³ C NMR (125 MHz, DMSO^) δ ppm 161.9, 160.7, 153.0, 141.0, 124.8, 83.8, 76.6, 69.8, 58.9, 52.3, 44.4, 32.7, 28.1, 26.2, 18.3, -4.1/-4.5.

Step B: methyl 2-[[3-[tert-butyl(dimethyl)silyl]oxy-4-methoxybutyl]amino]thiazole-4carboxylate.

Using the general HFIP deprotection procedure, starting with the product from Step A as the appropriate carbamate, 3.1 g (55%) of the desired product was obtained.

1H NMR (400 MHz, DMSO-d^ δ ppm 7.79 (t, 1H), 7.52 (s, 1H), 3.91 (m, 1H), 3.74 (s, 3H), 3.29-3.21 (m, 7H ), 1.75(m, 1H), 1.63 (m, 1H), 0.86 (s, 9H), 0.04 (s, 6H).

Step C: methyl 2-[[3-[tert-butyl(dimethyl)silyl]oxy-4-methoxybutyl]-[5-methyl-6-[^)-[3-(2trimethylsilylethoxymethyl)-1,3-benzothiazol- 2-ylidene]amino]pyridazin-3-yl]amino]thiazol-4carboxylate.

Using the general Buchwald reaction procedure II, starting from the product from step B and the product of synthesis 4a as a suitable halide, 118 mg (61%) of the desired product were obtained.

1H NMR (500 MHz, DMSO-d^ δ ppm 7.99 (s, 1H), 7.84 (d, 1H), 7.58 (s, 1H), 7.47 (d, 1H), 7.44 (td, 1H), 7.25 (td, 1H), 5.87 (s, 2H), 4.52/4.44 (m+m, 2H), 4.02 (m, 1H), 3.81 (s, 3H), 3.72 (t, 2H), 3.37 (m, 2H), 3.29 (s, 3H), 2.46 (s, 3H), 1.93/1.81 (m+m, 2H), 0.92 (t, 2H), 0.89 (s, 9H), 0.09/0.07 (s+s, 6H), -0.12 (s, 9H); ¹³ C NMR (125 MHz, DMSO-d^ δ ppm 162.3, 160.4, 157.7, 155.3, 150.9, 140.5, 137.5, 137.2, 127.2, 125.4, 123.5, 1 23.2 , 123.2, 117.5, 112.0, 76.7, 72.9, 69.6, 66.7, 58.9, 52.2, 44.8, 31.8, 26.3, 17.9, 17.8, -1.0, -4.1/-4.5; LC-MS-ESI (m/z): [ M+H] ⁺ , calculated for C ₃₄ H ₅₃ N ₆ O ₅ S ₂ Si ₂ : 745; found: 745.

Step D: 2-[[6-(1,3-benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]-(3-hydroxy-4methoxybutyl)amino]thiazol-4-carboxylic acid.

Using the general deprotection and hydrolysis procedure followed by repurification by reverse phase preparative chromatography (C18, 0.1% TFA in water:MeCN), the TFA salt of the desired product was obtained from the product from step C.

MSHR-ERI (m/z): [M+H] ⁺ , calculated for C21H23N6O4S2: 487.1217; found: 487.1222.

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Example 94: 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)-5-(1methanesulfonylpiperidin-4-yl)-1, 3-thiazole-4-carboxylic acid

Step A: ethyl 5-(1-methanesulfonylpiperidin-4-yl)-2-[methyl(5-methyl-6-{[(2Z)-3-{[2(trimethylsilyl)ethoxy]methyl}-2,3- dihydro-1,3-benzothiazol-2-ylidene]amino}pyridazin-3-yl)amino]1,3-thiazol-4-carboxylate.

To a stirred solution of synthesis product 5k (150 mg, 0.23 mmol, 1 eq.) and triethylamine (0.07 ml, 0.47 mmol, 2 eq.) in dichloromethane (5 ml) was added methanesulfonyl chloride (0.02 ml, 0.28 mmol, 1.2 eq.) with followed by the addition of a catalytic amount of 4-(dimethylamino)pyridine, and the mixture was stirred at ambient temperature overnight. The reaction mixture was diluted with dichloromethane, then washed with water followed by brine, dried (magnesium sulfate) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 12 g RediSep™ silica gel cartridge) eluting with a gradient of 0-60% ethyl acetate in isoheptane gave the title product as a white foam (106 mg, 0.15 mmol, 63%).

LC/MS (C ₃₁ H ₄₃ N ₇ O ₅ SiS ₃ ) 718 [M+H]+; RT 1.45 (LCMS-V-BI).

¹ H NMR (400 MHz, DMSOHO δ 7.89 (dd, J=7.9, 1.1 Hz, 1H), 7.68 (d, J=1.1 Hz, 1H), 7.51-7.39 (m, 2H), 7.29-7.21 (m, 1H), 5.87 (s, 2H), 4.31 (q, J=7.1 Hz, 2H), 3.77 (s, 3H), 3.76-3.63 (m, 5H), 2.93 (s, 3H), 2.89-2.78 (m , 2H), 2.46 (d, J=1.0 Hz, 3h), 2.12 (d, J=12.3 Hz, 2H), 1.78-1.63 (m, 2H), 1.33 (t, J=7.1 Hz, 3H), 0.97 -0.88 (m, 2H), -0.11 (s, 9H).

Step B: ethyl 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)-5-(1methanesulfonylpiperidin-4-yl)-1 ,3-thiazole-4-carboxylate.

Trifluoroacetic acid (1 mL, 13.4 mmol, 91 eq.) was added to a stirred solution of the product from Step A (106 mg, 0.15 mmol, 1 eq.) in dichloromethane (5 mL) and the mixture was stirred at ambient temperature overnight. The reaction mixture was partitioned between dichloromethane and saturated aqueous sodium bicarbonate, and the organic phase was washed with brine, dried (magnesium sulfate) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 12 g RediSep™ silica gel cartridge) eluting with a gradient of 0-5% methanol in dichloromethane resulted in a solid which was triturated with diethyl ether, filtered and dried in vacuo to give the title product as a yellow solid (75 mg, 0.13 mmol, 86%).

LC/MS (C ₂₅ H ₂₉ N7O ₄ S ₃ ) 588 [M+H]+; RT 1.36 (LCMS-V-BI).

1H NMR (400 MHz, DMSO-de) δ 7.95 (br s, 1h), 7.68 (s, 1H), 7.53 (br s, 1H), 7.38 (t, J=7.6 Hz, 1H), 7.20 (t, J=7.5 Hz, 1H), 4.30 (q, J=7.1 Hz, 2H), 3.80-3.67 (m, 6H), 2.93 (s, 3H), 2.84 (dd, J=13.0, 10.6 Hz, 2H), 2.46 (s, 3H), 2.12 (d, J=11.9 Hz, 2H), 1.79-1.59 (m, 2H), 1.33 (t, J=7.1 Hz, 3H).

Step C: 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl} (methyl)amino)-5 -(1methanesulfonylpiperidin-4-yl)-1, 3-thiazole-4-carboxylic acid.

To a solution of the product from step B (75 mg, 0.13 mmol, 1 eq.) in 1,4-dioxane (3 ml) was added lithium hydroxide monohydrate (26.8 mg, 0.64 mmol, 5 eq.), and the mixture was refluxed at during the night. The reaction mixture was allowed to cool to ambient temperature and the solids were collected by filtration and washed with 1,4-dioxane. Purification by reverse phase automated flash chromatography (CombiFlash Rf, C18 13 g RediSep column) eluting with a gradient of 5-95% acetonitrile in water gave the title product as a yellow solid (37.1 mg, 0.07 mmol, 52%).

HRMS-ERI (m/z) [M+H]+, calculated for C ₂₃ H ₂₆ N ₇ O ₄ S ₃ : 560.1203; found: 560.1209.

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Example 95: 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)-5-(1phenylmethanesulfonylpiperidin-4-yl)-1, 3-thiazole-4-carboxylic acid

Step A: ethyl 2-[methyl(5-methyl-6-{[(2Z)-3-{[2-(Ίrimethylsilyl)ethoxy]methyl}-2,3-dihydro-1,3benzothiazol-2-ylidene] amino } pyridazin-3 -yl)amino]-5-(1-phenylmethanesulfonylpiperidin-4-yl)-1,3thiazol-4-carboxylate.

To a stirred solution of synthesis 5k and triethylamine (0.07 ml, 0.47 mmol, 2 eq.) in dichloromethane (5 ml) was added alpha-toluenesulfonyl chloride (53.6 mg, 0.28 mmol, 1.2 eq.) followed by the addition of a catalytic amount of 4-(dimethylamino) pyridine, and the mixture was stirred at ambient temperature overnight. The reaction mixture was diluted with dichloromethane, washed with water followed by brine, dried (magnesium sulfate) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 12 g RediSep™ silica gel cartridge) eluting with a gradient of 0-60% ethyl acetate in isoheptane gave the title product as a creamy foam (95 mg, 0.12 mmol, 51%).

LC/MS (C ₃₇ H ₄₇ N ₇ O ₅ SiS ₃ ) 794 [M+H]+; RT 1.50 (LCMS-V-BI).

¹ H NMR (400 MHz, DMSO-06) δ 7.81 (dd, J=7.8, 1.1 Hz, 1H), 7.67 (d, J=1.1 Hz, 1H), 7.53-7.36 (m, 7H), 7.27-7.20 (m, 1H), 5.86 (s, 2H), 4.49 (s, 2H), 4.29 (q, J=7.1 Hz, 2H), 3.77 (s, 3H), 3.77-3.64 (m, 5H), 2.82 ( td, J=12.4, 2.3 Hz, 2H), 2.46 (d, J=1.0 Hz, 3H), 2.03-1.95 (m, 2H), 1.62-1.46 (m, 2H), 1.32 (t, J=7.1 Hz , 3H), 0.92 (dd, J=8.6, 7.4 Hz, 2H), -0.11 (s, 9H).

Step B: ethyl 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)-5-(1phenylmethanesulfonylpiperidin-4-yl)-1 ,3-thiazole-4-carboxylate.

Trifluoroacetic acid (0.5 mL, 6.71 mmol, 56 eq.) was added to a stirred solution of the product from Step A (95 mg, 0.12 mmol, 1 eq.) in dichloromethane (5 mL), and the mixture was stirred at ambient temperature overnight. The reaction mixture was partitioned between dichloromethane and saturated aqueous sodium bicarbonate, and the organic phase was washed with brine, dried (magnesium sulfate) and concentrated in vacuo. The resulting solid was triturated in diethyl ether, filtered and dried in vacuo to give the title product as a yellow solid (74 mg, 0.11 mmol, 93%).

LC/MS (C ₃₁ H ₃₃ N ₇ O ₄ S ₃ ) 664 [M+H] ⁺ ; RT 1.27 (LCMS-V-BI).

1H NMR (400 MHz, DMSOL) δ 11.03 (br s, 1H), 7.91 (br s, 1H), 7.74-7.65 (m, 1H), 7.53-7.33 (m, 7H), 7.25-7.13 (m, 1H ), 4.49 (s, 2H), 4.29 (q, J=7.1 Hz, 2H), 3.76 (s, 3H), 3.74-3.64 (m, 3H), 2.88-2.77 (m, 2H), 2.502.44 ( m, 3H), 2.03-1.94 (m, 2H), 1.60-1.46 (m, 2H), 1.32 (t, J=7.1 Hz, 3H).

Step C: 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)-5-(1phenylmethanesulfonylpiperidin-4-yl)-1, 3-thiazole-4-carboxylic acid.

To a solution of the product from step B (74 mg, 0.11 mmol, 1 eq.) in 1,4-dioxane (3 ml) was added lithium hydroxide monohydrate (23.4 mg, 0.56 mmol, 5 eq.) and the mixture was refluxed at for 6.5 h. The reaction mixture was allowed to cool to ambient temperature and purified by reverse phase flash chromatography (CombiFlash Rf, C18 13 g RediSep column) eluting with a gradient of 5-95% acetonitrile in water to give the desired product as yellow solid (43.2 mg, 0.07 mmol, 61%).

HRMS-ERI (m/z) [M+H] ⁺ , calculated for C ₂₉ H ₃₀ N ₇ O ₄ S ₃ : 636.1516; found: 636.1521.

Example 96: 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)-5-[1-(2phenylacetyl)piperidin-4- yl]-1,3-thiazol-4-carboxylic acid

Step A: ethyl 2-[methyl(5-methyl-6-{[(2Z)-3-{[2-(Ίrimethylsilyl)ethoxy]methyl}-2,3-dihydro-1,3

- 169 046336 benzothiazol-2-ylidene]amino}pyridazin-3-yl)amino]-5-[1-(2-phenylacetyl)piperidin-4-yl]-1,3-thiazol4-carboxylate.

Phenylacetyl chloride (0.04 ml, 0.28 mmol, 1.2 eq.) was added to a stirred solution of synthesis product 5k (150 mg, 0.23 mmol, 1 eq.) and triethylamine (0.07 ml, 0.47 mmol, 2 eq.) in dichloromethane (5 ml). and the mixture was stirred at ambient temperature overnight. The reaction mixture was diluted with dichloromethane, then washed with saturated aqueous sodium bicarbonate followed by brine, dried (magnesium sulfate) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 12 g RediSep™ silica gel cartridge) eluting with a gradient of 0-60% ethyl acetate in isoheptane gave the title product as an off-white foam (122 mg, 0.16 mmol, 69% ).

LC/MS (C ₃₈ H4 ₇ N ₇ O ₄ SiS ₂ ) 758 [M+H]+; RT 1.49 (LCMS-V-Bi).

¹ HNMR (400 MHz, DMSO-de) δ 7.88-7.81 (m, 1H), 7.66 (d, J=1.1 Hz, 1H), 7.51-7.41 (m, 2H), 7.39-7.32 (m, 2H), 7.30-7.20 (m, 4H), 5.87 (s, 2H), 4.59 (d, J=12.7 Hz, 1H), 4.29 (q, J=7.1 Hz, 2H), 4.10 (d, J=13.6 Hz, 1H ), 3.94-3.66 (m, 8H), 3.09 (t, J=12.7 Hz, 1H), 2.72-2.57 (m, 1H), 2.46 (d, J=1.0 Hz, 3H), 2.11-1.83 (m, 2H), 1.50-1.36 (m, 2H), 1.32 (t, J=7.1 Hz, 3H), 0.97-0.87 (m, 2H), -0.11 (s, 9H).

Step B: ethyl 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)-5-[1-(2phenylacetyl)piperidin-4 -yl]-1,3-thiazol-4-carboxylate.

Trifluoroacetic acid (0.5 mL, 6.71 mmol, 42 eq.) was added to a stirred solution of the product from Step A (122 mg, 0.16 mmol, 1 eq.) in dichloromethane (5 mL), and the mixture was stirred at ambient temperature overnight. The reaction mixture was partitioned between dichloromethane and saturated aqueous sodium bicarbonate, and the organic phase was washed with brine, dried (magnesium sulfate) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 12 g RediSep™ silica gel cartridge) eluting with a gradient of 0-5% methanol in dichloromethane provided the title product as a yellow solid (81.8 mg, 0.13 mmol, 81%) .

LC/MS (C ₃₂ H ₃₃ N ₇ O ₃ S ₂ ) 628 [M+H] ⁺ ; RT 1.22 (LCMS-V-Bi).

1H NMR (400 MHz, DMSO) δ 11.06 (br s, 1H), 7.98 (br s, 1H), 7.71-7.65 (m, 2H), 7.43-7.32 (m, 3H), 7.31-7.14 (m, 4H ), 4.59 (d, J=13.3 Hz, 1H), 4.29 (q, J=7.1 Hz, 2H), 4.10 (d, J=13.9 Hz, 1H), 3.91-3.70 (m, 6H), 3.09 (t , J=12.4 Hz, 1H), 2.72-2.58 (m, 1H), 2.48-2.43 (m, 3H), 2.07 (d, J=12.6 Hz, 1H), 1.91 (d, J=13.0 Hz, 1H) , 1.50-1.33 (m, 2H), 1.32 (t, J=7.1 Hz, 3H).

Step C: 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)-5-[1-(2phenylacetyl)piperidin-4- yl]-1,3-thiazol-4-carboxylic acid.

To a solution of the product from step B (81.8 mg, 0.13 mmol, 1 eq.) in 1,4-dioxane (3 ml) was added lithium hydroxide monohydrate (27.3 mg, 0.65 mmol, 5 eq.), and the mixture was refluxed at during the night. The reaction mixture was allowed to cool to ambient temperature and purified by reverse phase auto flash chromatography (CombiFlash Rf, C18 13 g RediSep column) eluting with a gradient of 5-95% acetonitrile in water to give the title product as a yellow solid ( 54.3 mg, 0.09 mmol, 70%).

HRMS-ERI (m/z) [M+H] ⁺ , calculated for C ₃₀ H ₃₀ N ₇ O ₃ S ₂ : 600.1846; found: 600.1850.

Example 97: 5-(1-acetylpiperidin-4-yl)-2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3yl}(methyl)amino)-1, 3-thiazole-4-carboxylic acid

Step A: ethyl 5-(1-acetylpiperidin-4-yl)-2-[methyl(5-methyl-6-{[(2Z)-3-{[2(trimethylsilyl)ethoxy]methyl}-2,3- dihydro-1,3-benzothiazol-2-ylidene]amino}pyridazin-3-yl)amino]1,3-thiazol-4-carboxylate.

To a stirred solution of synthesis 5k and triethylamine (0.07 mL, 0.47 mmol, 2 eq.) in dichloromethane (5 mL) was added acetyl chloride (0.02 mL, 0.28 mmol, 1.2 eq.) and the mixture was stirred at ambient temperature overnight. The reaction mixture was diluted with dichloromethane, then washed with saturated aqueous sodium bicarbonate followed by brine, dried (magnesium sulfate) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 12 g RediSep™ silica gel cartridge) eluting with a gradient of 0-100% ethyl acetate in isoheptane gave the title product as a colorless foam (119 mg, 0.17 mmol, 74%).

LC/MS (C ₃₂ H4 ₃ N ₇ O ₄ SiS ₂ ) 682 [M+H] ⁺ ; RT 1.44 (LCMS-V-Bi).

- 170 046336

1H NMR (400 MHz, DMSO-dg) δ 7.90-7.83 (m, 1H), 7.67 (d, J=1.2 Hz, 1H), 7.50-7.39 (m, 2H), 7.29-7.21 (m, 1H), 5.86 (s, 2H), 4.57 (d, J=13.3 Hz, 1H), 4.31 (q, J=7.1 Hz, 2H), 3.96 (d, J=13.6 Hz, 1H), 3.91-3.82 (m, 1H ), 3.77 (s, 3H), 3.75-3.67 (m, 2H), 3.14 (t, J=12.2 Hz, 1H), 2.69-2.54 (m, 1H), 2.45 (d, 3H), 2.06 (s, 3H), 2.061.95 (m, 2H), 1.72-1.56 (m, 1H), 1.54-1.38 (m, 1H), 1.33 (t, J=7.1 Hz, 3H), 0.96-0.87 (m, 2H) , -0.11 (s, 9H).

Step B: ethyl 5-(1-acetylpiperidin-4-yl)-2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin3 -yl}(methyl)amino)-1 ,3-thiazole-4-carboxylate.

To a stirred solution of the product from step A (119 mg, 0.17 mmol, 1 eq.) in tetrahydrofuran (5 ml) was added tetrabutylammonium fluoride (1 M in tetrahydrofuran; 0.07 ml, 0.5 mmol, 3 eq.) and ethylenediamine (0.03 ml, 0.5 mmol, 3 eq.), and the mixture was heated at 60°C overnight. The reaction mixture was allowed to cool to ambient temperature, then partitioned between dichloromethane and saturated aqueous sodium bicarbonate, and the organic phase was washed with brine, dried (magnesium sulfate) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 12 g RediSep™ silica gel cartridge) eluting with a gradient of 0-4% methanol in dichloromethane provided the title product as a yellow solid (40 mg, 0.07 mmol, 42%) .

LC/MS (C ₂₆ H ₂₉ N ₇ O ₃ S ₂ ) 552 [M+H]+; RT 1.09 (LCMS-V-Bi).

1H NMR (400 MHz, DMSO^ ₆ ) δ 11.08 (br s, 1H), 7.94 (br s, 1H), 7.69 (s, 1H), 7.57 (br s, 1H), 7.39 (t, J=7.5 Hz , 1H), 7.23 (t, 1H), 4.58 (d, J=13.3 Hz, 1H), 4.31 (q, J=7.1 Hz, 2H), 4.02-3.93 (m, 1H), 3.92-3.82 (m, 1H), 3.77 (s, 3H), 3.21-3.09 (m, 1H), 2.66-2.55 (m, 1H), 2.47 (s, 3H), 2.07 (s, 3H), 2.06-1.96 (m, 2H) , 1.76-1.57 (m, 1H), 1.54-1.39 (m, 1H), 1.34 (t, J=7.1 Hz, 3H).

Step C: 5-(1-acetylpiperidin-4-yl)-2-({ 6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3 yl}(methyl)amino)-1 ,3-thiazole-4-carboxylic acid.

To a solution of the product from step B (40 mg, 0.07 mmol, 1 eq.) in 1,4-dioxane (3 ml) was added lithium hydroxide monohydrate (15.2 mg, 0.36 mmol, 5 eq.), and the mixture was refluxed at during the night. The reaction mixture was allowed to cool to ambient temperature and purified by reverse phase auto flash chromatography (CombiFlash Rf, C18 13 g RediSep column) eluting with a gradient of 5-95% acetonitrile in water to give the title product as a yellow solid ( 32.4 mg, 0.06 mmol, 85%).

HRMS-ERI (m/z) [M+H]+, calculated for C ₂₄ H ₂₆ N ₇ O ₃ S ₂ : 524.1533; found: 524.1541.

Example 98: 2-[[6-(1,3-Benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]-(4-hydroxy-3 methoxybutyl)amino]thiazol-4-carboxylic acid

Step A: Methyl 2-[tert-butoxycarbonyl-[4-[tert-butyl(dimethyl)silyl]oxy-3methoxybutyl]amino]thiazole-4-carboxylate.

Using the general Mitsunobu reaction procedure, starting with methyl 2-(tert-butoxycarbonylamino)thiazole-4-carboxylate and synthesis product 2i as starting materials and carrying out the reaction in THF at room temperature. within 3 hours, 4.54 g (86%) of the target product were obtained.

LC-MS-ESI (m/z): [M+H]+, calculated for C ₂ iH ₃₉ N ₂ O ₆ SSi: 475; found: 475.

Step B: methyl 2-[[4-[tert-butyl(dimethyl)silyl]oxy-3-methoxybutyl]amino]thiazole-4carboxylate.

Using the general HFIP deprotection procedure, starting with the product from Step A as the appropriate carbamate, 2.88 g (55%) of the desired product was obtained.

1H NMR (400 MHz, DMSO^ ₆ ) δ ppm 7.80 (t, 1H), 7.52 (s, 1H), 3.74 (s, 3H), 3.59 (m, 2H), 3.32 (s, 3H), 3.28 (m, 3H), 1.76 (m, 1H), 1.61 (m, 1H), 0.86 (s, 9H), 0.04 (s, 6H).

Step C: methyl 2-[[4-[tert-butyl(dimethyl)silyl]oxy-3-methoxybutyl]-[5-methyl-6-[^)-[3-(2trimethylsilylethoxymethyl)-1,3-benzothiazol- 2-ylidene]amino]pyridazin-3-yl]amino]thiazol-4carboxylate.

Using the general Buchwald reaction procedure II, starting from the product from step B and the product of synthesis 4a as a suitable halide, 319 mg (70%) of the desired product were obtained.

1H NMR (500 MHz, DUSSH) δ ppm 7.99 (s, 1H), 7.85 (dm, 1H), 7.66 (s, 1H), 7.48 (dm, 1H), 7.44 (m, 1H), 7.26 (m, 1H), 5.88 (s, 2H), 4.60 (m, 1H), 4.47 (m, 1H), 3.81 (s, 3H), 3.72 (m, 2H), 3.66 (dd, 1H), 3.53 (dd, 1H), 3.29 (s, 3H), 3.27 ( m, 1H), 2.46 (s, 3H), 2.03 (m, 1H), 1.76 (m, 1H), 0.92 (m, 2H), 0.77 (s, 9H), 0.00 (s, 3H), -0.01 ( s, 3H), -0.12 (s, 9H).

Step D: 2-[[6-(1,3-benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]-(4-hydroxy-3

- 171 046336 methoxybutyl)amino]thiazole-4-carboxylic acid.

Using the general deprotection and hydrolysis procedure followed by repurification by reverse phase preparative chromatography (C18, 0.1% TFA in water:MeCN), the TFA salt of the desired product was obtained from the product from step C.

MSHR-ERI (m/z): [M+H]+, calculated for C ₂₁ H ₂₃ N ₆ O ₄ S: 487.1217; found: 487.1222.

Example 99: 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)-5-(1methylpiperidin-4-yl)-1, 3-thiazole-4-carboxylic acid

Step A: ethyl 2-[methyl(5-methyl-6-{[(2Z)-3-{[2-(trimethylsilyl)ethoxy]methyl}-2,3-dihydro-1,3benzothiazol-2-ylidene]amino }pyridazin-3-yl)amino]-5-(1-methylpiperidin-4-yl)-1,3-thiazol-4carboxylate.

Formaldehyde (37% in water; 0.04 ml, 0.47 mmol, 2 eq.), sodium triacetoxyborohydride was added to a stirred solution of synthesis product 5k (150 mg, 0.23 mmol, 1 eq.) in a 1:1 mixture of acetonitrile/dichloromethane (10 ml). (149 mg, 0.7 mmol, 3 eq.) and glacial acetic acid (10 μl), and the mixture was stirred at ambient temperature overnight. The reaction mixture was partitioned between dichloromethane and saturated aqueous sodium bicarbonate, and the organic phase was washed with brine, dried (magnesium sulfate) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 12 g RediSep™ silica gel cartridge) eluting with a gradient of 0-100% ethyl acetate in isoheptane gave the title product as an off-white foam (76.7 mg, 0.12 mmol, 50% ).

LC/MS (C ₃₁ H ₄₃ N ₇ O ₃ SiS ₂ ) 654 [M+H]+; RT 1.24 (LCMS-V-BI).

¹ H NMR (400 MHz, DMSO-d^ δ 7.88 (dd, J=7.7, 1.0 Hz, 1H), 7.67 (d, J=1.2 Hz, 1H), 7.51-7.39 (m, 2H), 7.28-7.22 (m, 1H), 5.87 (s, 2H), 4.29 (q, J=7.1 Hz, 2H), 3.76 (s, 3H), 3.75-3.67 (m, 2H), 3.64-3.52 (m, 1H), 2.89 (d, J=11.1 Hz, 2H), 2.46 (d, J=1.0 Hz, 3H), 2.21 (s, 3H), 2.01-1.90 (m, 4H), 1.75-1.61 (m, 2H), 1.32 (t, J=7.1 Hz, 3H), 0.99-0.88 (m, 2H), -0.11 (s, 9H).

Step B: ethyl 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)-5-(1methylpiperidin-4-yl)-1 ,3-thiazole-4-carboxylate.

Trifluoroacetic acid (1.0 mL, 13.4 mmol, 114 eq.) was added to a stirred solution of the product from Step A (76.7 mg, 0.12 mmol, 1 eq.) in dichloromethane (4 mL), and the mixture was stirred at ambient temperature overnight. The reaction mixture was partitioned between dichloromethane and saturated aqueous sodium bicarbonate, and the organic phase was washed with brine, dried (magnesium sulfate) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 12 g RediSep™ silica gel cartridge) eluting with a gradient of 0-10% methanol in dichloromethane gave the title product as a yellow glass (24.1 mg, 0.05 mmol, 39%) .

LC/MS (C ₂₅ H ₂₉ N ₇ O ₂ S ₂ ) 524 [M+H]+; RT 0.87 (LCMS-V-BI).

1H NMR (400 MHz, DMSO^6) δ 8.01-7.90 (m, 1H), 7.69 (s, 1H), 7.54 (br s, 1H), 7.44-7.35 (m, 1H), 7.22 (t, J= 7.7 Hz, 1H), 4.30 (q, J=7.1 Hz, 2H), 3.76 (s, 3H), 3.64-3.53 (m, 1H), 2.97-2.86 (m, 2H), 2.47 (s, 3h), 2.22 (s, 3H), 2.04-1.89 (m, 4H), 1.77-1.60 (m, 2H), 1.33 (t, J=7.1 Hz, 3H).

Step C: 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)-5-(1methylpiperidin-4-yl)-1, 3-thiazole-4-carboxylic acid.

To a solution of the product from step B (24 mg, 0.05 mmol, 1 eq.) in 1,4-dioxane (3 ml) was added lithium hydroxide monohydrate (9.62 mg, 0.23 mmol, 5 eq.), and the mixture was refluxed at during the night. The reaction mixture was allowed to cool to ambient temperature and concentrated in vacuo. Purification by reverse phase automated flash chromatography (CombiFlash Rf, C18 13 g RediSep column) eluting with a gradient of 5-95% acetonitrile in water gave the title product as a yellow solid (17.2 mg, 0.03 mmol, 76% ).

HRMS-ERI (m/z) [M+H]+, calculated for C ₂₃ H ₂₆ N ₇ O ₂ S ₂ : 496.1584; found: 496.1602.

- 172 046336

Example 100: 2-({6-[( 1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl } (methyl)amino)-5-{ 3 -[2fluoro-4-(3 -hydroxyprop-1-yn-1-yl)phenoxy]propyl}-1,3-thiazol-4-carboxylic acid

Step A: ethyl 5-[3-(2-fluoro-4-{3-[(4-methoxyphenyl)methoxy]prop-1-yn-1-yl}phenoxy)propyl]-2[methyl(5-methyl- 6-{[(2Z)-3-{[2-(trimethylsilyl)ethoxy]methyl}-2,3-dihydro-1,3-benzothiazol-2ylidene]amino}pyridazin-3-yl)amino]-1,3 -thiazole-4-carboxylate.

To a solution of synthesis product 5g (249 mg, 0.34 mmol, 1 eq.) in dimethylformamide (5 ml) was added synthesis product 6k (128 mg, 0.45 mmol, 1.3 eq.) and cesium carbonate (336 mg, 1.03 mmol, 3 eq. ), and the mixture was heated at 80°C overnight. The reaction mixture was partitioned between ethyl acetate and water, and the organic phase was washed with brine, dried (magnesium sulfate) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 12 g RediSep™ silica gel cartridge) eluting with a gradient of 0-50% ethyl acetate in isoheptane gave the title product as a yellow gum (231 mg, 0.26 mmol, 76%).

LC/MS (C45H51FN6O6SiS2) 883 [M+H]+; RT 1.41 (ZHMS-U-V2).

¹ H NMR (400 MHz, DMSO^) δ 7.84 (d, 1H), 7.67 (d, J=1.1 Hz, 1H), 7.49-7.40 (m, 2H), 7.37 (dd, J=11.9, 2.0 Hz, 1H), 7.31-7.22 (m, 4H), 7.21-7.14 (m, 1H), 6.94-6.89 (m, 2H), 5.86 (s, 2H), 5.87 (s, 2H), 4.48 (s, 2H) , 4.32 (s, 2H), 4.26 (t, 2H), 4.16 (t, J=6.1 Hz, 2H), 3.78 (s, 3H), 3.75 (s, 3H), 3.75-3.68 (m, 2H), 3.34-3.22 (m, 2H), 2.46 (d, J=1.0 Hz, 3H), 2.19-2.08 (m, 2H), 1.30 (t, 3H), 0.97-0.88 (m, 2H), -0.11 (s , 9H).

Step B: ethyl 5-{3-[2-fluoro-4-(3-hydroxyprop-1-yn-1-yl)phenoxy]propyl}-2-[methyl(5-methyl-6{[(2Z)- 3 - {[2-(trimethylsilyl)ethoxy]methyl }-2,3-dihydro-1,3-benzothiazol-2-ylidene] amino } pyridazin-3 yl)amino]-1,3-thiazol-4-carboxylate.

Trifluoroacetic acid (1.8 ml) was added to a solution of the product from step A (391 mg, 0.44 mmol, 1 eq.) in dichloromethane (18 ml), and the mixture was stirred at ambient temperature for 1 hour. The reaction mixture was diluted with dichloromethane, cooled to 0°C, and neutralized by adding 0.5 M aqueous sodium hydroxide solution. The layers were separated and the organic phase was dried (magnesium sulfate) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 12 g RediSep™ silica gel cartridge) eluting with a gradient of 0-50% ethyl acetate in isoheptane gave the title product as an off-white solid (121 mg, 0.16 mmol, 36%) .

LC/MS (C ₃₇ H ₄₃ FN ₆ O ₅ SiS ₂ ) 763 [M+H] ⁺ ; RT 1.29 (ZHMS-U-V2).

1H NMR (400 MHz, DMSOM ₆ ) δ 7.87-7.81 (m, 1H), 7.67 (s, 1H), 7.50-7.40 (m, 2H), 7.33-7.13 (m, 4H), 5.86 (s, 2H) , 5.30 (t, J=6.0 Hz, 1H), 4.34-4.21 (m, 4H), 4.15 (t, J=6.2 Hz, 2H), 3.77 (s, 3H), 3.76-3.67 (m, 2H), 3.32-3.22 (m, 2H), 2.46 (d, J=1.0 Hz, 3H), 2.20-2.06 (m, 2H), 1.28 (t, 3H), 0.97-0.88 (m, 2H), -0.12 (s , 9H).

Step C: ethyl 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)-5-{3-[2fluoro-4-( 3 -hydroxyprop-1 -yn-1 -yl)phenoxy]propyl}-1,3-thiazol-4-carboxylate.

To a solution of the product from step B (121 mg, 0.16 mmol, 1 eq.) in tetrahydrofuran (15 ml) was added ethylenediamine (31.8 μl, 0.48 mmol, 3 eq.) and TBAF (476 μl, 0.48 mmol, 3 eq.), and the mixture was heated at 65°C overnight. The reaction mixture was allowed to cool to ambient temperature, then partitioned between ethyl acetate and water, dried (magnesium sulfate) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 12 g RediSep™ silica gel cartridge) eluting with a gradient of 0-100% ethyl acetate in isoheptane provided the title product as a yellow solid (43 mg, 0.07 mmol, 43%) .

LC/MS (C ₃₁ H ₂₉ FN ₆ O ₄ S ₂ ) 633 [M+H] ⁺ ; RT 2.43 (LCMS-VC).

1H NMR (400 MHz, DMSO^) δ 11.08 (br s, 1H), 7.92 (br s, 1H), 7.67 (br s + s, 2H), 7.39 (t, J=7.5 Hz, 1H), 7.29 ( dd, J=11.9, 1.9 Hz, 1H), 7.25-7.12 (m, 2H), 5.29 (t, J=5.9 Hz, 1H), 4.33-4.20 (m, 4H), 4.14 (t, 2H), 3.77 (s, 3H), 3.31-3.21 (m, 1H), 2.46 (s, 3H), 2.20-2.07 (m, 2H), 1.29 (t, J=7.1 Hz, 3H).

Step D: 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)-5-{3-[2fluoro-4-(3 -hydroxyprop-1-yn-1-yl)phenoxy]propyl}-1,3-thiazol-4-carboxylic acid.

To a solution of the product from step C (50 mg, 0.08 mmol, 1 eq.) in 1,4-dioxane (15 ml) was added lithium hydroxide monohydrate (33.2 mg, 0.79 mmol, 10 eq.) and the mixture was refluxed at during the night. The reaction mixture was allowed to cool to ambient temperature and concentrated in vacuo. Purification using reverse phase automated flash chromatography (Com

- 173 046336 biFlash Rf, C18 13 g RediSep column) eluting with a gradient of 30-95% acetonitrile in water gave the title product as a yellow solid (31.9 mg, 0.05 mmol, 67%).

MSVR-ERI (m/z) [MH] ^- , calculated for C ₂₉ H ₂₄ FN ₆ O ₄ S2: 603.1290; found: 603.1294.

Example 101: 3-[3-[4-[3-[2-[[6-(1,3-benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]methylamino]-4carboxythiazol-5-yl] propoxy]-3-fluorophenyl]prop-2-ynyldimethylammonio]propane-1-sulfonate.

Using a general alkylation, deprotection and hydrolysis procedure, starting from the compound of Example 50, Step A, and oxathiolane 2,2-dioxide, the desired product was obtained.

MSVR-ERI (m/z): [M+H]+, calculated for C ₃₄ H ₃₇ FN ₇ O ₆ S ₃ : 754.1946; found: 754.1947.

Example 102: 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl} (methyl)amino)-5cyclohexyl-1,3-thiazol-4-carboxylic acid

Stage A: cyclohexyl(iodo)zinc.

Zinc (3.03 g, 46.3 mmol, 3 eq.) and lithium chloride (0.98 g, 23.1 mmol, 1.5 eq.) were added to a 50 mL oven-dried Schlenk flask, and the mixture was heated at 160°C for 20 min on high vacuum, then allowed to cool to ambient temperature and placed under a nitrogen atmosphere on the Schlenk line. Dimethylacetamide (10 mL) was added followed by 1,2-dibromoethane (0.13 mL, 1.54 mmol, 0.1 eq.) and the mixture was stirred for 20 min. Iodocyclohexane (2 mL, 15.4 mmol, 1 eq) was added slowly and the mixture was then heated at 40°C overnight. The reaction mixture was allowed to cool to ambient temperature and cannulated through a filter (cotton wadding/celite/cotton wadding) under low vacuum in a dry 25 ml Schlenk flask to obtain the desired product as a 1 M solution (as determined by titration with 0.5 M iodine solution ), which was used without further characterization.

Step B: ethyl 5-cyclohexyl-2-[methyl(5-methyl-6-{[(2Z)-3-{[2-(trimethylsilyl)ethoxy]methyl}-2,3dihydro-1,3-benzothiazol-2 -ylidene]amino}pyridazin-3-yl)amino]-1,3-thiazol-4-carboxylate.

A solution of synthesis product 11b (200 mg, 0.31 mmol, 1 eq.) and copper iodide(!) (112 mg, 0.63 mmol, 2 eq.) were loaded into a flask dried in an oven. Dimethylacetamide (5 ml) was added and the mixture was stirred for 5 min, after which the product from Step A (1 M in dimethyl acetamide; 1.42 ml, 1.42 mmol, 4.5 eq.) was added slowly and the mixture was stirred at ambient temperature overnight . The reaction mixture was poured onto saturated aqueous ammonium chloride, then extracted with dichloromethane (x2). The combined organic extracts were washed with brine, dried (magnesium sulfate) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 12 g RediSep™ silica gel cartridge) eluting with a gradient of 0-30% ethyl acetate in isoheptane gave the title product as a yellow gum (135 mg, 0.21 mmol, 67%).

LC/MS (C ₃₁ H ₄₂ N ₆ O ₃ S ₂ Si) 639 [M+H]+; RT 1.68 (LCMS-V-BI).

¹ H NMR (400 MHz, DMSO^ ₆ ) δ 7.85 (dd, J=7.6, 1.0 Hz, 1H), 7.66 (d, J=1.2 Hz, 1H), 7.50-7.40 (m, 2H), 7.29-7.22 (m, 1H), 5.86 (s, 2H), 4.28 (q, J=7.1 Hz, 2H), 3.76 (s, 3H), 3.75-3.69 (m, 2H), 3.68-3.56 (m, 1H), 2.45 (s, 3H), 2.05-1.97 (m, 2H), 1.87-1.78 (m, 2H), 1.77-1.68 (m, 2H), 1.52-1.35 (m, 4H), 1.32 (t, 3H), 0.96-0.88 (m, 2H), -0.11 (s, 9H).

Step C: 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)-5cyclohexyl-1,3-thiazol-4-carboxylic acid .

Trifluoroacetic acid (1 ml, 13.4 mmol, 63.7 eq.) was added to the stirred solution

- 174 046336 product from step B (135 mg, 0.21 mmol, 1 eq.) in dichloromethane (6 ml), and the mixture was stirred at ambient temperature overnight. The reaction mixture was partitioned between dichloromethane and saturated aqueous sodium bicarbonate, and the organic phase was washed with brine, dried (magnesium sulfate) and concentrated in vacuo. The resulting solid was suspended in 1,4-dioxane (4 mL), then lithium hydroxide (44.2 mg, 1.05 mmol, 5 eq.) was added and the mixture was refluxed overnight. The reaction mixture was allowed to cool to ambient temperature and concentrated in vacuo. Purification by reverse phase automated flash chromatography (CombiFlash Rf, C18 13 g RediSep column) eluting with a gradient of 5-95% acetonitrile in water gave the title product as a yellow solid (42 mg, 0.09 mmol, 41% ).

HRMS-ERI (m/z) [M+H]+, calculated for C ₂₃ H ₂₅ N ₆ O ₂ S ₂ : 481.1475; found: 481.1518.

Example 103: 2-[[6-(1,3-Benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]-(2,3dihydroxypropyl)amino]thiazol-4-carboxylic acid

Step A: methyl 2-[tert-butoxycarbonyl-[(2,2-dimethyl-1,3-dioxolan-4-yl)methyl]amino]thiazole-4carboxylate.

Using the general Mitsunobu reaction procedure, starting with methyl 2-(tert-butoxycarbonylamino)thiazole-4-carboxylate and (2,2-dimethyl-1,3-dioxolan-4-yl)methanol as starting materials and carrying out the reaction in THF at .T. within 19 hours, 5.09 g (91%) of the target product was obtained.

¹ H NMR (500 MHz, DMSO-d^ δ ppm 8.10 (s, 1H), 4.40 (m, 1H), 4.26/4.07 (dd+dd, 2H), 4.01/3.79 (dd+dd, 2H ), 3.80 (s, 3H), 1.53 (s, 9H), 1.34 (s, 3H), 1.23 (s, 3H); ¹³ C NMR (125 MHz, DMSO-06) δ ppm 161.8, 161.1, 153.2, 140.9, 124.9, 73.1, 67.1, 52.4, 49.1, 28.1, 27.1, 25.8; MSHR-ERI (m/z): [M+H]+, calculated for C16H25N2O6S: 373.1433; found: 373.1431.

Step B: methyl 2-[(2,2-dimethyl-1,3-dioxolan-4-yl)methylamino]thiazol-4-carboxylate.

Using the general HFIP deprotection procedure, starting with the product from Step A as the appropriate carbamate, 1.77 g (48%) of the desired product was obtained.

1H NMR (500 MHz, DMSO-d^ δ ppm 7.94 (t, 1H), 7.53 (s,H), 4.23 (m, 1H), 4.00/3.66 (dd+dd, 2H), 3.74 (s , 3H), 3.39/3.35 (m+m, 2H), 1.34 (s, 3H), 1.26 (s, 3H); ¹³ C NMR (125 MHz, DMSO-dg) δ ppm 168.7, 162.0, 142.3 , 117.6, 74.3, 67.0, 52.1, 47.4, 27.3, 25.8; MSHR-ERI (m/z): [M+H]+, calculated for C11H17N2O4S: 273.0909; found: 273.0906.

Step C: methyl 2-[(2,2-dimethyl-1,3-dioxolan-4-yl)methyl-[5-methyl-6-[(7)-[3-(2trimethylsilylethoxymethyl)-1,3-benzothiazole -2-ylidene]amino]pyridazin-3-yl]amino]thiazol-4carboxylate.

Using the general Buchwald reaction procedure II, starting from the product from step B and the product of synthesis 4a as a suitable halide, 450 mg (33%) of the desired product were obtained.

1H NMR (500 MHz, DMSO-dg) δ ppm 8.00 (s, 1H), 7.84 (d, 1H), 7.83 (s, 1H), 7.47 (d, 1H), 7.44 (td, 1H), 7.25 (td, 1H), 5.87 (s, 2H), 4.60 (m, 2H), 4.55 (m, 1H), 4.09/3.83 (dd+dd, 2H), 3.82 (s, 3H), 3.71 (t, 2H), 2.44 (s, 3H), 1.27/1.19 (s +s, 6H), 0.92 (t, 2H), -0.12 (s, 9H); ¹³ C NMR (125 MHz, DMSO^6) δ ppm 162.2, 161.1, 157.7, 155.4, 151.7, 140.4, 137.5, 136.4, 127.2, 125.4, 123.5, 123.4, 123.2, 119.1, 112.0, 73.9, 72.8, 66.9, 66.7, 52.3, 50.2, 26.6/25.6, 17.8, 17.7, -1.0; MSVR-ERI (m/z): [M+H] ⁺ , calculated for SJ^: 643.2192; found: 643.2184.

Step D: 2-[[6-(1,3-benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]-(2,3-dihydroxypropyl)amino]thiazol-4-carboxylic acid.

Using the general deprotection and hydrolysis procedure followed by repurification by reverse phase preparative chromatography (C18, 25 mM NH4HCO3 water:MeSA), the desired product was obtained from the product from step C.

MSHR-ERI (m/z): [M+H] ⁺ , calculated for C ₁₉ H ₁₉ N6O ₄ S ₂ : 459.0909; found: 459.0891.

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Example 104: 2-[[6-(1,3-Benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]-(4,5-dihydroxypentyl)amino]thiazol-4-carboxylic acid he

Step A: Methyl 2-[tert-butoxycarbonyl(pent-4-enyl)amino]thiazole-4-carboxylate.

Using the general Mitsunobu reaction procedure, starting with methyl 2-(tert-butoxycarbonylamino)thiazol-4-carboxylate and 4-penten-1-ol as starting materials and carrying out the reaction in THF at room temperature. within 2 hours, 4.36 g (89%) of the target product was obtained.

Step B: Methyl 2-[3-(2,2-dimethyl-1,3-dioxolan-4-yl)propylamino]thiazole-4-carboxylate.

To 4.35 g of the product from step A (13.3 mmol) was added 3.6 g of 4-methyl-4-oxidomorpholin-4ium hydrate (26.6 mmol, 2 eq.), 135 mg of osmium tetroxide (2.5 wt.% in 2-methylpropan-2-ol , 0.013 mmol, 0.001 eq.), 10 ml of water and 100 ml of 2-methylpropan-2-ol, and the mixture was stirred at room temperature. for 18 hours. After adding 50 ml of 1 M Na ₂ S ₂ O ₃ and removing tBuOH under reduced pressure, the mixture was extracted with DCM and the combined organic phases were dried and concentrated to give 6.25 g (130%) methyl 2-[tert- butoxycarbonyl(4,5-dihydroxypentyl)amino]thiazole-4-carboxylate. The crude product was added to 100 ml of EtOH, treated with 20 ml of 1N. HCl at 100°C for 24 h, and concentrated to give 5.42 g (157%) methyl 2-(4,5-dihydroxypentylamino)thiazole-4-carboxylate (LC-MS-ESI (m/z): [M+ H]+, calculated for C ₁₀ H ₁₇ N ₂ O ₄ S: 261; found: 261). The product was then added to 50 ml of 2,2dimethoxypropane and 124 mg of 4-methylbenzenesulfonic acid (0.72 mmol, 0.05 eq.) was added and the mixture was stirred at reflux for 3 hours. After concentration, the residue was added to DCM and washed with conc. NaHCO ₃ to obtain 3.66 g (92% in three stages) of the target product.

1H NMR (500 MHz, DMSO-06) δ ppm 7.81 (t, 1H), 7.50 (s, 1H), 4.04 (m, 1H), 3.98/3.42 (dd+dd, 2H), 3.73 (s, 3H), 3.23 (q, 2H), 1.54 (m, 2H), 1.54 (m, 2H), 1.30 (s, 3H), 1.25 (s, 3H); ¹³ C NMR (125 MHz, DMSOC) δ ppm 117.0, 75.6, 69.0, 52.0, 44.7, 27.3, 27.3, 26.1, 25.5; MSVR-ERI (m/z): [M+H] ⁺ , calculated for C13H21N2O4S: 301.1222; found: 301.1217.

Step C: methyl 2-[3-(2,2-dimethyl-1,3-dioxolan-4-yl)propyl-[5-methyl-6-[^)-[3-(2trimethylsilylethoxymethyl)-1,3- benzothiazol-2-ylidene]amino]pyridazin-3-yl]amino]thiazol-4carboxylate.

Using the general Buchwald reaction procedure II, starting from the product from step B and the product of synthesis 4a as a suitable halide, 74 mg (50%) of the desired product were obtained.

1H NMR (500 MHz, DMSO-06) δ ppm 7.99 (s, 1H), 7.84 (d, 1H), 7.72 (brs., 1H), 7.47 (d, 1H), 7.44 (td, 1H), 7.25 (td, 1H), 5.87 (s, 2H), 4.49/4.39 (m+m, 2H), 4.13 (m, 1H), 4.02/3.44 (dd+t, 2H), 3.82 (s, 3H), 3.72 (m, 2H), 2.47 (s, 3H), 1.79/1.73 (m+m, 2H), 1.67-1.52 (m, 2H), 1.29/1.26 (s+s, 6H), 0.92 (m, 2H), -0.11 (s, 9H); ¹³ C NMR (125 MHz, DMSO-06) δ ppm 162.3, 157.7, 155.3, 150.9, 137.5, 137.2, 127.2, 125.4, 123.5, 123.2, 123.2, 117.7, 112.0, 75.5, 72.9, 69.1, 66.7, 52.3, 47 .3, 30.3, 27.4/26.1, 23.7, 17.8, 17.8, -1.0; MSHR-ERI (m/z): [M+H] ⁺ , calculated for C ₃₁ H43N ₆ O ₅ S ₂ Si: 671.2505; found: 671.2489.

Step D: 2-[[6-(1,3-benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]-(4,5dihydroxypentyl)amino]thiazol-4-carboxylic acid.

Using the general deprotection and hydrolysis procedure followed by repurification by reverse phase preparative chromatography (C18, 25 mM NH4HCO3 water:MeCN), the desired product was obtained from the product from step C.

MSHR-ERI (m/z): [M+H]+, calculated for C ₂₁ iH ₂₃ N ₆ O ₄ S ₂ : 487.1222; found: 487.1212.

Example 105: 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)-5cyclopentyl-1,3-thiazol-4-carboxylic acid

Stage A: bromine(cyclopentyl)zinc.

Zinc (1.83 g, 28 mmol, 3 equiv.) and lithium chloride (593 mg, 14 mmol, 1.5 equiv.) were loaded into an oven-dried Schlenk flask (50 ml). A vacuum was created in the flask and the mixture was heated at 160°C in

- 176 046336 for 20 minutes, then allowed to cool to ambient temperature and placed under a nitrogen atmosphere on the Schlenk line. Dimethylacetamide (10 mL) was added followed by 1,2 dibromoethane (0.08 mL, 0.93 mmol, 0.1 eq.) and the mixture was stirred for 20 min. Bromocyclopentane (1 mL, 9.33 mmol, 1 eq.) was added slowly and the reaction mixture was then heated at 40°C overnight. The reaction mixture was allowed to cool to ambient temperature and cannulated through a filter (cotton wadding/celite/cotton wadding) under low vacuum in a dry 25 ml Schlenk flask to obtain the desired product as a 0.25 M solution (as determined by titration with 0.5 M iodine solution ), which was used without further characterization.

Step B: ethyl 5-cyclopentyl-2-[methyl(5-methyl-6-{[(2Z)-3-{[2-(trimethylsilyl)ethoxy]methyl}-2,3dihydro-1,3-benzothiazol-2 -ylidene]amino}pyridazin-3-yl)amino]-1,3-thiazol-4-carboxylate.

Synthesis product 11b (200 mg, 0.31 mmol, 1 eq.) and copper(D) iodide (120 mg, 0.63 mmol, 2 eq.) were loaded into a flask dried in an oven, followed by the addition of dimethylacetamide (5 ml), and the mixture was stirred for 5 min, after which the product from step A (0.25 M in dimethylacetamide; 5.66 ml, 1.42 mmol, 4.5 eq.) was added slowly and the mixture was stirred overnight. The reaction mixture was poured onto a saturated aqueous ammonium chloride solution and extracted with dichloromethane (x2). The organic phase was washed with brine, dried (magnesium sulfate) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 12 g RediSep™ silica gel cartridge) eluting with a gradient of 0-50% ethyl acetate in isoheptane gave a solid which was triturated with isoheptane, filtered, washed with diethyl ether and dried in vacuo to obtain the target product as a white solid (112 mg, 0.18 mmol, 57%).

LC/MS (C30H ₄ 0NeO3S2Si) 625 [M+H]+; RT 1.79 (LCMS-V-BI).

1H NMR (400 MHz, DMSO-dg) δ 7.88-7.80 (m, 1H), 7.66 (d, J=1.1 Hz, 1H), 7.52-7.39 (m, 2H), 7.29-7.21 (m, 1H), 5.86 (s, 2H), 4.29 (q, J=7.1 Hz, 2H), 4.06-3.88 (m, 1H), 3.75 (s, 3H), 3.74-3.68 (m, 2H), 2.45 (d, J= 1.0 Hz, 3H), 2.24-2.12 (m, 2H), 1.90-1.76 (m, 2H), 1.73-1.49 (m, 2H), 1.31 (t, J=7.1 Hz, 3H), 0.96-0.81 (m , 2H), 0.11 (s, 9H).

Step C: 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)-5cyclopentyl-1,3-thiazol-4-carboxylic acid .

Trifluoroacetic acid (0.8 mL, 10.73 mmol, 60 eq.) was added to a stirred solution of the product from Step B (112 mg, 0.18 mmol, 1 eq.) in dichloromethane (4 mL), and the mixture was stirred at ambient temperature overnight. The reaction mixture was partitioned between dichloromethane and saturated aqueous sodium bicarbonate, and the organic phase was washed with brine, dried (magnesium sulfate) and concentrated in vacuo. The resulting solid was suspended in 1,4-dioxane (3 mL), then lithium hydroxide (37.6 mg, 0.9 mmol, 5 eq.) was added and the mixture was refluxed overnight. The reaction mixture was allowed to cool to ambient temperature and concentrated in vacuo. Purification by reverse phase automated flash chromatography (CombiFlash Rf, C18 13 g RediSep column) eluting with a gradient of 5-95% acetonitrile in water gave the title product as a brown solid (34.8 mg, 0.07 mmol, 42% ).

HRMS-ERI (m/z) [M—H]—, calculated for C ₂₂ H ₂₁ N ₆ O ₂ S ₂ : 465.1173; found: 465.1175.

Example 106: 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)-5-ethenyl1,3-thiazol-4-carboxylic acid

Step A: ethyl 5-ethenyl-2-[methyl(5-methyl-6-{[(2Z)-3-{[2-(trimethylsilyl)ethoxy]methyl}-2,3dihydro-1,3-benzothiazol-2 -ylidene]amino}pyridazin-3-yl)amino]-1,3-thiazol-4-carboxylate.

Pd(dppf) _Cl2.CH2Cl2 ( _12.9 mg, 0.02 mmol, 0.1 eq.) was added to a solution of synthesis product 11b (100 mg, 0.16 mmol, 1 equiv.) in a 5:1 _{tetrahydrofuran} /water mixture (6 ml). ), potassium vinyl trifluoroborate (31.6 mg, 0.24 mmol, 1.5 eq.) and potassium carbonate (65.2 mg, 0.47 mmol, 3 eq.), and the mixture was refluxed for 3 days. The reaction mixture was allowed to cool to ambient temperature, then partitioned between ethyl acetate and water, and the organic phase was washed with brine, dried (magnesium sulfate) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 12 g RediSep™ silica gel cartridge) eluting with a gradient of 0-50% ethyl acetate in isoheptane provided the title product as a white solid (39.5 mg, 0.07 mmol, 43%) .

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LC/MS (C ₂₇ H ₃₄ N ₆ O ₃ S2Si) 583 [M+H]+; RT 1.52 (LCMS-V-BI).

1H NMR (400 MHz, DMSO^ ₆ ) δ 7.88-7.80 (m, 1H), 7.71 (d, J=1.1 Hz, 1H), 7.59-7.40 (m, 3H), 7.28-7.23 (m, 1H), 5.87 (s, 2H), 5.71-5.62 (m, 1H), 5.45-5.37 (m, 1H), 4.32 (q, J=7.1 Hz, 2H), 3.80 (s, 3H), 3.78-3.68 (m, 2H), 2.46 (d, J=1.0 Hz, 3H), 1.33 (t, J=7.1 Hz, 3H), 0.97-0.87 (m, 2H), -0.11 (s, 9H).

Step B: 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl} (methyl)amino)-5-ethenyl-1,3thiazol-4-carboxylic acid .

Trifluoroacetic acid (0.5 mL, 6.71 mmol, 99 eq.) was added to a stirred solution of the product from Step A (39.5 mg, 0.07 mmol, 1 eq.) in dichloromethane (3 mL), and the mixture was stirred at ambient temperature overnight. The reaction mixture was partitioned between dichloromethane and saturated aqueous sodium bicarbonate, and the organic phase was washed with brine, dried (magnesium sulfate) and concentrated in vacuo. The resulting solid was suspended in 1,4-dioxane (3 mL), lithium hydroxide monohydrate (14.2 mg, 0.34 mmol, 5 eq.) was added and the mixture was refluxed overnight. The reaction mixture was allowed to cool to ambient temperature and concentrated in vacuo. Purification by reverse phase automated flash chromatography (CombiFlash Rf, C18 13 g RediSep column) eluting with a gradient of 5-95% acetonitrile in water gave the title product as a yellow solid (13.4 mg, 0.03 mmol, 47% ).

MSVR-ERI (m/z) [MH] ^- , calculated for C ₁₉ H ₁₅ N ₆ O ₂ S ₂ : 423.0703; found: 423.0711.

Example 107: 2-[[6-(1,3-Benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]-(3-hydroxy-2morpholinopropyl)amino]thiazol-4-carboxylic acid

Step A: Methyl 2-[tert-butoxycarbonyl-[3-[tert-butyl(dimethyl)silyl]oxy-2morpholinopropyl]amino]thiazole-4-carboxylate.

Using the general Mitsunobu reaction procedure, starting with methyl 2-(tert-butoxycarbonylamino)thiazole-4-carboxylate and synthesis product 2k as starting materials and carrying out the reaction at room temperature. within 1 hour, 1.29 g (89%) of the target product was obtained.

1H NMR (500 MHz, DMSO^) δ ppm 8.08 (s, 1H), 4.36/3.86 (dd+dd, 2H), 3.78 (s, 3H), 3.77/3.67 (dd+dd, 2H), 3.32/3.21 (m+m, 4H), 3.01 (m , 1H), 2.82/2.36 (m+m, 4H), 1.53 (s, 9H), 0.86 (s, 9H), 0.04/0.03 (s, 6H); ¹³ C NMR (125 MHz, DMSOC) δ ppm 161.8, 161.6, 153.3, 140.7, 124.5, 67.4, 63.8, 61.0, 52.3, 50.1, 45.8, 28.2, 26.2, 18.3, -5.1; IR: 2954, 1703; MSHR-ERI (m/z): [M+H] ⁺ , calculated for C ₂₃ H ₄₃ N ₃ O ₆ SSi: 516.2564; found: 516.2570.

Step B: methyl 2-[[3-[tert-butyl(dimethyl)silyl]oxy-2-morpholinopropyl]amino]thiazole-4carboxylate.

Using the general HFIP deprotection procedure, starting with the product from Step A as the appropriate carbamate, 0.71 g (68%) of the desired product was obtained.

1H NMR (500 MHz, DMSO-dg) δ ppm 7.58 (t, 1H), 7.51 (s, 1H), 3.74/3.66 (dd+dd, 2H), 3.73 (s, 3H), 3.593.45 (m, 4H), 3.32/3.29 (m+m, 2H ), 2.71 (m, 1H), 2.70/2.56 (m+m, 4H), 0.87 (s, 9H), 0.04 (s, 6H); ¹³ C NMR (125 MHz, DMSO-dg) δ ppm 168.8, 162.0, 142.5, 117.1, 67.3, 64.5, 61.2, 52.0, 50.1, 43.2, 26.3, 18.3, -5.0; IR: 2854, 1729; MSHR-ERI (m/z): [M-C4H9] ⁺ , calculated for C14H ₂₄ N ₃ O ₄ SSi: 358.1262; found: 358.1255.

Step C: methyl 2-[[3-[tert-butyl(dimethyl)silyl]oxy-2-morpholinopropyl]-[5-methyl-6-[^)-[3-(2trimethylsilylethoxymethyl)-1,3-benzothiazol- 2-ylidene]amino]pyridazin-3-yl]amino]thiazol-4carboxylate.

Using the general Buchwald reaction procedure II, starting from the product from step B and the product of synthesis 4a as the appropriate halide, followed by purification by reverse phase preparative chromatography (C18, 25 mM NH4HCO3 water:MeC^, gave 270 mg (70%) target product.

1H NMR (500 MHz, DMSO^ ₆ ) δ ppm 7.97 (s, 1H), 7.84 (d, 1H), 7.74 (s, 1H), 7.48 (d, 1H), 7.44 (t, 1H), 7.26 (t, 1H), 5.89 (s, 2H), 4.71 /4.26 (dd+dd, 2H), 3.84/3.79 (dd+dd, 2H), 3.80 (s, 3H), 3.71 (t, 2H), 3.27/3.16 (t+t, 4H), 2.97 (m, 1H), 2.88/2.39 (t+t, 4H), 2.45 (s, 3H), 0.92 (t, 2H), 0.88 (s, 9H), 0.06 (s, 6H), -0.12 (s, 9H); ¹³ C NMR (125 MHz, DMSO^) δ ppm 127.2, 123.5, 123.2, 122.9, 118.5, 112.0, 72.8, 67.3, 66.7, 64.2, 61.0, 52.3, 50.3, 46.4, 26.3, 17.8, 17.8, -1, -5.1; MSVR-ERI (m/z): [M+H] ⁺ , calculated for CзgH5gN7O5S2Si2:

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786.3322; found: 786.3312.

Step D: 2-[[6-(1,3-benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]-(3-hydroxy-2morpholinopropyl)amino]thiazol-4-carboxylic acid.

Using the general deprotection and hydrolysis procedure followed by repurification by reverse phase preparative chromatography (C18, 25 mM NH4HCO3 water:MeCN), the desired product was obtained from the product from step C.

HRMS-ERI (m/z): [M+H]+, calculated for C ₂₃ H ₂₆ N ₇ O ₄ S ₂ : 528.1487; found: 528.1489.

Example 108: 2-[[6-(1,3-benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]-(3,4dihydroxybutyl)amino]-5-[3-[2-fluoro-4- [3-(methylamino)prop-1-ynyl]phenoxy]propyl]thiazole-4carboxylic acid

Step A: methyl 5-[3-[4-[3-[tert-butoxycarbonyl(methyl)amino]prop-1-ynyl]-2fluorophenoxy]propyl]-2-[2-(2,2-dimethyl-1, 3-dioxolan-4-yl)ethyl-[5-methyl-6-[^)-[3-(2trimethylsilylethoxymethyl)-1,3-benzothiazol-2-ylidene] amino]pyridazin-3 -yl] amino]thiazol- 4carboxylate.

Using the general Buchwald reaction procedure II, starting from 350 mg of synthesis product 3h (0.57 mmol, 1 eq.) and 235 mg of synthesis product 4a (0.57 mmol, 1 eq.) as a suitable halide, 490 mg (87%) of the target product were obtained product.

¹ H NMR (500 MHz, DMSO-d^ δ ppm 7.84 (d, 1H), 7.68 (s, 1H), 7.47 (d, 1H), 7.44 (td, 1H), 7.32 (brd., 1H ), 7.25 (td, 1H), 7.22 (d, 1H), 7.16 (t, 1H), 5.86 (s, 2H), 4.49/4.33 (m+m, 2H), 4.20 (br., 2H), 4.17 (m, 1H), 4.15 (t, 2H), 4.04/3.63 (dd+dd, 2H), 3.77 (s, 3H), 3.72 (t, 2H), 3.27 (t, 2H), 2.84 (br., 3H), 2.45 (s, 3H), 2.13 (m, 2H), 1.75 (m, 2H), 1.40 (s, 9H), 1.37/1.24 (s+s, 6H), 0.92 (t, 2H), - 0.11 (s, 9H); ¹³ C NMR (125 MHz, DMSO-d,) δ ppm 129.1, 127.2, 123.5, 123.2, 119.3, 117.5, 115.5, 112.0, 108.6, 73.7, 72.8, 68.9, 68.4, 66.7, 51.9, 44.4, 38.5, 33.8, 30.9, 28.5, 27.3/26.0, 23.3, 23.1, 17.9, 17.8, -1.0; HRMS-ERI (m/z): [M+H] ⁺ , calculated for C ₄₈ H ₆₃ FN ₇ O ₈ S ₂ Si: 976.3927; found: 976.3916.

Step B: 2-[[6-(1,3-benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]-(3,4dihydroxybutyl)amino]-5-[3-[2-fluoro-4- [3-(methylamino)prop-1-ynyl]phenoxy]propyl]thiazole-4carboxylic acid.

Using a general deprotection and hydrolysis procedure, starting with the product from Step A as the appropriate methyl ester, the desired product was obtained.

MSHR-ERI (m/z): [M+H] ⁺ , calculated for C ₃₃ H ₃₅ FN ₇ O ₅ S ₂ : 692.2120; found: 692.2114.

Example 109: 2-[[6-(1,3-Benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]-(3-hydroxy-4methoxybutyl)amino]-5-[3-[2-fluoro- 4-[3-(methylamino)prop-1-ynyl]phenoxy]propyl]thiazole-4-carboxylic acid

Step A: methyl 5-[3-[4-[3-[tert-butoxycarbonyl(methyl)amino]prop-1-ynyl]-2fluorophenoxy]propyl]-2-[[3-[tert-butyl(dimethyl)silyl ]oxy-4-methoxybutyl]-[5-methyl-6-[^)-[3-(2trimethylsilylethoxymethyl)-1,3-benzothiazol-2-ylidene] amino]pyridazin-3 -yl] amino]thiazol-4carboxylate.

Using the general Buchwald reaction procedure II, starting from 320 mg of synthesis product 31 (0.46 mmol, 1 eq.) and 188 mg of synthesis product 4a (0.46 mmol, 1 eq.) as a suitable halide, 415 mg (84%) of the target product were obtained product.

- 179 046336

1H NMR (500 MHz, DMSO-de) δ ppm 7.84 (d, 1H), 7.55 (s, 1H), 7.47 (d, 1H), 7.44 (td, 1H), 7.32 (brd., 1H), 7.25 (td, 1H), 7.21 (d, 1H), 7.16 (t, 1H), 5.86 (s, 2H), 4.49/4.37 (m+m, 2H), 4.20 (br., 2H), 4.14 (t, 2H), 4.01 (m, 1H), 3.76 (s , 3H), 3.71 (t, 2H), 3.36 (m, 2H), 3.28 (s, 3H), 3.27 (t, 2H), 2.84 (br., 3H), 2.45 (s, 3H), 2.12 (m , 2H), 1.90/1.79 (m+m, 2H), 1.40 (s, 9H), 0.92 (t, 2H), 0.88 (s, 9H), 0.08/0.06 (s/s, 6H), -0.12 ( s, 9H); ¹³ C NMR (125 MHz, DMSO-de) δ ppm 129.1, 127.2, 123.4, 123.2, 119.3, 117.3, 115.5, 112.0, 76.6, 72.8, 69.6, 68.4, 66.7, 58.8, 51.9, 44.2, 38.5, 33.8, 31.8, 3 0.9, 28.5, 26.3, 23.1, 17.8, 17.8, -1.0, -4.1/-4.5; MSHR-ERI (m/z): [M+H] ⁺ , calculated for C ₅₂ H ₇₅ FN ₇ O ₈ S ₂ Si ₂ : 1064.4636; found: 1064.4629.

Step B: 2-[[6-(1,3-benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]-(3-hydroxy-4methoxybutyl)amino]-5-[3-[2-fluoro- 4-[3-(methylamino)prop-1-ynyl]phenoxy]propyl]thiazole-4-carboxylic acid.

Using a general deprotection and hydrolysis procedure, starting with the product from Step A as the appropriate methyl ester, the desired product was obtained.

MSHR-ERI (m/z): [M+H] ⁺ , calculated for C ₃₄ H ₃₇ FN ₇ O ₅ S ₂ : 706.2276; found: 706.2285.

Example 110: 2-[[6-(1,3-Benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]-(4hydroxybutyl)amino]thiazole-4-carboxylic acid

Step A: methyl 2-[tert-butoxycarbonyl-[4-[tert-butyl(dimethyl)silyl]oxybutyl]amino]thiazole 4-carboxylate.

Using the general Mitsunobu reaction procedure, starting with methyl 2-(tert-butoxycarbonylamino)thiazole-4-carboxylate and 4-[tert-butyl(dimethyl)silyl]oxybutan-1-ol as starting materials and carrying out the reaction in THF at RT . within 1 hour, 3.38 g (98%) of the target product was obtained.

1H NMR (500 MHz, DMSO-dg) δ ppm 8.09 (s, 1H), 4.07 (t, 2H), 3.79 (s, 3H), 3.59 (t, 2H), 1.69 (qn, 2H), 1.53 (s, 9H), 1.45 (qn, 2H), 0.83 (s, 9H), 0.00 (s, 6H); ¹³ C NMR (125 MHz, DMSO-de) δ ppm 161.9, 160.9, 153.1, 141.0, 124.8, 83.7, 62.5, 52.4, 47, 30.0, 28.1, 26.3, 24.5, 18.3, -4.9; MSHR-ERI (m/z): [M+H] ⁺ , calculated for C ₂₀ H ₃₇ N ₂ O ₅ SSi: 445.2192; found: 445.2193.

Step B: methyl 2-[tert-butoxycarbonyl-[4-[tert-butyl(dimethyl)silyl]oxybutyl]amino]thiazole 4-carboxylate.

Using the general HFIP deprotection procedure, starting with the product from Step A as the appropriate carbamate, 2.0 g (80%) of the desired product was obtained.

1H NMR (500 MHz, DMSO-de) δ ppm 7.79 (t, 1H), 7.50 (s, 1H), 3.73 (s, 3H), 3.59 (t, 2H), 3.22 (q, 2H), 1.58 (m, 2H), 1.51 (m, 2H), 0.85 (s, 9H), 0.02 (s, 6H); ¹³ C NMR (125 MHz, DMSO-de) δ ppm 168.7, 162.0, 142.6, 116.9, 62.7, 52.0, 44.7, 30.2, 26.3, 25.6, -4.8; IR: 2929, 1732; MSHR-ERI (m/z): [M+H] ⁺ , calculated for C ₁₅ H ₂₉ N ₂ O ₃ SSi: 345.1668; found: 345.1652.

Step C: methyl 2-[4-[tert-butyl(dimethyl)silyl]oxybutyl-[5-methyl-6-[(7)-[3-(2trimethylsilylethoxymethyl)-1,3-benzothiazol-2-ylidene]amino ]pyridazin-3-yl]amino]thiazol-4carboxylate.

Using the general Buchwald reaction procedure II, starting from the product from step B and the product of synthesis 4a as a suitable halide, 451 mg (73%) of the desired product were obtained.

1H NMR (500 MHz, DMSO-de) δ ppm 7.98 (s, 1H), 7.84 (dm, 1H), 7.69 (s, 1H), 7.47 (dm, 1H), 7.44 (m, 1H), 7.25 (m, 1H), 5.87 (s, 2H), 4.47 (t, 2H), 3.82 (s, 3H), 3.72 (m, 2H), 3.62 (t, 2H), 2.46 (s, 3H), 1.76 (m, 2H), 1.53 (m, 2H), 0.92 ( m, 2H), 0.80 (s, 9H), -0.01 (s, 6H), -0.10 (s, 9H); ¹³ C NMR (125 MHz, DMSO-de) δ ppm 127.2, 123.5, 123.2, 123.1, 117.7, 112.0, 72.9, 66.7, 62.5, 52.2, 47.2, 29.7, 26.2, 23.8, 17.8, 17.8, -1.0, -4.9; IR: 2951, 1732; MSHR-ERI (m/z): [M+H] ⁺ , calculated for C ₃₃ H ₅₁ N ₆ O ₄ S ₂ Si ₂ : 715.2952; found: 715.2963.

Step D: 2-[[6-(1,3-benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]-(4-hydroxybutyl)amino]thiazole-4-carboxylic acid.

Using the general deprotection and hydrolysis procedure followed by repurification by reverse phase preparative chromatography (C18, 25 mM NH ₄ HCO ₃ water:MeCN), the desired product was obtained from the product from step C.

MSHR-ERI (m/z): [M+H] ⁺ , calculated for C ₂₀ H ₂₁ N ₆ O ₃ S ₂ : 457.1117; found: 457.1108.

- 180 046336

Example 111: 2-[[6-(1,3-Benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]-(3-hydroxy-2methoxypropyl)amino]thiazol-4-carboxylic acid

Step A: Methyl 2-[tert-butoxycarbonyl-[3-[tert-butyl(dimethyl)silyl]oxy-2methoxypropyl]amino]thiazole-4-carboxylate.

Using the general Mitsunobu reaction procedure starting with methyl 2-(tert-butoxycarbonylamino)thiazole-4-carboxylate and product.

Synthesis of 2j as starting materials and carrying out the reaction in THF at room temperature. within 1 hour, 683 mg (88%) of the target product were obtained.

¹ H NMR (400 MHz, DMSO-dg) δ ppm 8.09 (s, 1H), 4.22 (dd, 1H), 4.06 (dd, 1H), 3.79 (s, 3H), 3.71-3.55 (m, 3H), 3.29 (s, 3H), 1.52 (s, 9H) , 0.84 (s, 9H), 0.01 (s, 3H), 0.00 (s, 3H).

Step B: methyl 2-[[3-[tert-butyl(dimethyl)silyl]oxy-2-methoxypropyl]amino]thiazole-4carboxylate.

Using the general HFIP deprotection procedure, starting with the product from Step A as the appropriate carbamate, 514 mg (quantity) of the desired product was obtained.

¹ H NMR (400 MHz, DMSO^6) δ ppm 7.80 (t, 1H), 7.52 (s, 1H), 3.73 (s, 3H), 3.34 (s, 3H), 3.66 (dd, 1H), 3.62 (dd, 1H), 3.41 (m, 1H), 3.39 (m, 1H), 3.26 (m, 1h), 0.86 (s, 9H), 0.04 (s, 3H), 0.03 (s, 3h); LC-MS-ESI (m/z): [M+H]+, calculated for C ₁₅ H ₂₉ N ₂ O ₄ SSi: 361; found: 361.

Step C: methyl 2-[[3-[tert-butyl(dimethyl)silyl]oxy-2-methoxypropyl]-[5-methyl-6-[^)-[3-(2trimethylsilylethoxymethyl)-1,3-benzothiazol- 2-ylidene]amino]pyridazin-3-yl]amino]thiazol-4carboxylate.

Using the general Buchwald reaction procedure II, starting from the product from step B and the product of synthesis 4a as a suitable halide, 313 mg (70%) of the desired product were obtained.

1H NMR (500 MHz, DMSO-dg) δ ppm 7.98 (s, 1H), 7.84 (d, 1H), 7.75 (s, 1H), 7.51-7.41 (m, 2H), 7.26 (tm, 1H), 5.88 (s, 1H), 5.87 (s, 1H) , 4.53 (dd, 1H), 4.41 (dd, 1H), 3.81 (s, 3H), 3.78 (m, 2H), 3.75-3.65 (m, 3H), 3.29 (s, 3H), 2.44 (s, 3H ), 0.92 (t, 2H), 0.86 (s, 9h), 0.05 (s, 3H), 0.04 (s, 3H), -0.12 (s, 9H).

Step D: 2-[[6-(1,3-benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]-(3-hydroxy-2methoxypropyl)amino]thiazol-4-carboxylic acid.

Using the general procedure of deprotection and hydrolysis, followed by re-purification using reverse-phase preparative chromatography (C18, 0.1% TFA in water-MeC), the TFA salt of the target product was obtained from the product from step C.

MSHR-ERI (m/z): [M+H] ⁺ , calculated for C ₂₀ H ₂₁ N ₆ O ₄ S ₂ : 473.1066; found: 473.1063.

Example 112: 2-[[6-(1,3-benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]-(5-hydroxy-4methoxypentyl)amino]-5-[3-[2-fluoro- 4-[3-(methylamino)prop-1-ynyl]phenoxy]propyl]thiazole-4carboxylic acid

Step A: methyl 5-[3-[4-[3-[tert-butoxycarbonyl(methyl)amino]prop-1-ynyl]-2fluorophenoxy]propyl]-2-[[5-[tert-butyl(dimethyl)silyl ]oxy-4-methoxypentyl]-[5-methyl-6-[^)-[3-(2trimethylsilylethoxymethyl)-1,3-benzothiazol-2-ylidene]amino]pyridazin-3-yl]amino]thiazol-4carboxylate.

Using the general Buchwald reaction procedure II, starting with 12 g of synthesis 3f (13 mmol) and 6.30 g of synthesis 4a (15.6 mmol) as the appropriate halide, 14 g (83%) of the desired product were obtained.

Step B: 2-[[6-(1,3-benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]-(5-hydroxy-4methoxypentyl)amino]-5-[3-[2-fluoro- 4-[3-(methylamino)prop-1-ynyl]phenoxy]propyl]thiazole-4

- 181 046336 carboxylic acid.

Using a general deprotection and hydrolysis procedure, starting with the product from Step A as the appropriate methyl ester, the desired product was obtained.

1H NMR (500 MHz, DMSO^) δ ppm 7.91 (d, 1H), 7.66 (s, 1H), 7.53 (d, 1H), 7.39 (td, 1H), 7.26 (dd, 1H), 7.21 (td, 1H), 7.18 (dd, 1H), 7.15 (t, 1H), 4.38 (m, 2H), 4.14 (t, 2H), 3.47 (s, 2H), 3.41/3.36 (dd+dd, 2H), 3.28 (s, 3H), 3.26 (t, 2H ), 3.20 (m, 1H), 2.46 (s, 3H), 2.31 (s, 3H), 2.13 (m, 2H), 1.72 (m, 2H), 1.57/1.49 (m+m, 2H); ¹³ C NMR (125 MHz, DMSO-06) δ ppm 128.9, 126.6, 122.6, 122.3, 119.1, 118.3, 116.9, 115.5, 81.4, 68.5, 62.7, 57.1, 47.0, 40.2, 35.2, 31.0, 28.2, 23.3, 23.1, 1 7.8 MSVR-ERI (m/z): [M +H] ⁺ , calculated for C ₃₅ H ₃₉ FN ₇ O ₅ S ₂ : 720.2433; found: 720.2437.

Example 113: 2-[[6-(1,3-Benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]-(4-hydroxy-5morpholinopentyl)amino]-5-[3-[2-fluoro- 4-[3-(methylamino)prop-1-ynyl]phenoxy]propyl]thiazole-4carboxylic acid

Step A: methyl 5-[3-[4-[3-[tert-butoxycarbonyl(methyl)amino]prop-1-ynyl]-2fluorophenoxy]propyl]-2-[[4-[tert-butyl(diphenyl)silyl ]oxy-5-morpholinopentyl]-[5-methyl-6-[^)-[3-(2trimethylsilylethoxymethyl)-1,3-benzothiazol-2-ylidene] amino]pyridazin-3 -yl]amino]thiazol-4carboxylate.

Using the general tosylate alkylation procedure, starting from the synthesis product 5a and morpholine as a suitable amine, the desired product was obtained.

Step B: 2-[[6-(1,3-benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]-(4-hydroxy-5 morpholinopentyl)amino]-5-[3-[2-fluoro -4-[3-(methylamino)prop-1-ynyl]phenoxy]propyl]thiazole-4carboxylic acid.

Using a general deprotection and hydrolysis procedure, starting with the product from Step A as the appropriate methyl ester, the desired product was obtained.

MSVR-ERI (m/z): [M+H] ⁺ , calculated for Q8H44FN8O5S2: 775.2855; found: 775.2851.

Example 114: 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(4hydroxybutyl)amino)-5-(3-{2-fluoro-4- [3-(methylamino)prop-1-yn-1-yl]phenoxy}propyl)-1,3-thiazol-4carboxylic acid

Using the general deprotection and hydrolysis procedure starting from the appropriate methyl ester from Synthesis 5c, Step A, the desired product was obtained.

MSHR-ERI (m/z): [M+H] ⁺ , calculated for C ₃₃ H ₃₅ FN ₇ O ₄ S ₂ : 676.2170; found: 676.2177.

- 182 046336

Example 115: 2-[[6-(1,3-Benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]-(3-hydroxy-2morpholinopropyl)amino]-5-[3-[2-fluoro- 4-[3-(methylamino)prop-1-ynyl]phenoxy]propyl]thiazole-4carboxylic acid

ΗΝ _χ

Step A: methyl 5-[3-[4-[3-[tert-butoxycarbonyl(methyl)amino]prop-1-ynyl]-2fluorophenoxy]propyl]-2-[[3-[tert-butyl(dimethyl)silyl ]oxy-2-morpholinopropyl]-[5-methyl-6-[(2)-[3-(2trimethylsilylethoxymethyl)-1,3-benzothiazol-2-ylidene] amino]pyridazin-3-yl] amino]thiazol-4carboxylate .

Using the general Buchwald reaction procedure II, starting from 390 mg of synthesis product 3r (0.59 mmol, 1 eq.) and 216 mg of synthesis product 4a (0.59 mmol, 1 eq.) as a suitable halide, 500 mg (83%) of the target product were obtained product.

1H NMR (500 MHz, DMSO-dg) δ ppm 7.84 (dm, 1H), 7.71 (s, 1H), 7.47 (dm, 1H), 7.44 (m, 1H), 7.31 (brd, 1H), 7.25 (m, 1H), 7.21 (brd, 1H), 7.14 (t, 1H), 5.88/5.85 (d+d, 2H), 4.66/4.19 (dd+dd, 2H), 4.20 (brs, 2H), 4.14 (t, 2H), 3.82/3.77 (dd+dd, 2H), 3.75 (s, 3H), 3.71 (m, 2H), 3.29/3.26 (m+m, 2H), 3.28/3.18 (m+m, 4H), 2.95 (m, 1H), 2.89/2.39 ( m+m, 4H), 2.84 (brs, 3H), 2.43 (s, 3H), 2.13 (m, 2H), 1.40 (s, 9H), 0.92 (m, 2H), 0.87 (s, 9H), 0.05 (s, 6H), -0.12 (s, 9H); ¹³ C NMR (125 MHz, DMSO-dg) δ ppm 163.1, 157.4, 156.8/151.5, 141.2, 134.6, 129.0, 127.2, 123.4, 123.1, 119.2, 118.3, 115.4, 111.9, 85.1, 82.2, 72.7, 68.3, 67 .3, 66.6, 64.1, 60.9, 51.8, 50.1, 45.9, 38.5, 33.7, 30.9, 28.4, 26.2, 23.0, 17.9, 17.7, -1.1, -5.2; MSHR-ERI (m/z): [M+H] ⁺ , calculated for C ₅₄ H ₇₈ FN ₈ O ₈ S ₂ Si ₂ : 1105.4901; found: 1105.4896.

Step B: 2-[[6-(1,3-benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]-(3-hydroxy-2morpholinopropyl)amino]-5-[3-[2-fluoro- 4-[3-(methylamino)prop-1-ynyl]phenoxy]propyl]thiazole-4carboxylic acid.

Using a general deprotection and hydrolysis procedure, starting with the product from Step A as the appropriate methyl ester, the desired product was obtained.

MSHR-ERI (m/z): [M+H] ⁺ , calculated for C ₃₆ H ₄₀ FN ₈ O ₅ S ₂ : 747.2542; found: 747.2531.

Example 116: 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)-5-(3-{4-[3(dimethylamino )prop-1 -yn-1 -yl]-2-fluorophenoxy}-2-methylpropyl)-1,3-thiazol-4-carboxylic acid

•N\

Step A: ethyl 2-[methyl(5-methyl-6-{[(22)-3-{[2-(trimethylsilyl)ethoxy]methyl}-2,3-dihydro-1,3benzothiazol-2-ylidene]amino }pyridazin-3-yl)amino]-5-(2-methyl-3-oxopropyl)-1,3-thiazol-4carboxylate.

To a solution of synthesis product 11b (1.2 g, 1.89 mmol, 1 equiv.) in 1,4-dioxane (20 ml) was added 2methyl-2-propen-1-ol (0.48 ml, 5.66 mmol, 3 equiv.), H^ dicyclohexylmethylamine (0.81 ml, 3.78 mmol, 2 eq.) and 2-(di-tert-butylphosphino)-1-phenyl-1H-pyrrole (54.3 mg, 0.19 mmol, 0.1 eq.). The mixture was bubbled with nitrogen (10 min) and tris(dibenzylideneacetone)dipalladium(0) (86.4 mg, 0.09 mmol, 0.05 eq) was added and the mixture was heated at 100°C in a stoppered flask overnight. The reaction mixture was partitioned between ethyl acetate and water, and the organic phase was washed with brine, dried (magnesium sulfate) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 40 g RediSep™ silica gel cartridge) eluting with a gradient of 0-40% ethyl acetate in isoheptane gave the desired product as a yellow oil (779

- 183 046336 mg, 1.24 mmol, 66%).

LC/MS (C29H38N6O ₄ SiS2) 627 [M+H]+; RT 2.86 (LCMS-VC).

1H NMR (400 MHz, DMSOO δ 9.67 (d, J=1.4 Hz, 1H), 7.88-7.81 (m, 1H), 7.68-7.63 (m, 1H), 7.51-7.40 (m, 2H), 7.29-7.21 (m, 1H), 5.85 (s, 2H), 4.28 (q, 2H), 3.77 (s, 3H), 3.75-3.66 (m, 2H), 3.49 (dd, J=14.4, 6.7 Hz, 1H), 3.16 (dd, J=14.4, 7.6 Hz, 1H), 2.86-2.74 (m, 1H), 2.44 (d, 3H), 1.31 (t, 3H), 1.08 (d, 3H), 0.95-0.88 (m, 2H), -0.12 (s, 9H).

Step B: ethyl 5-(3-hydroxy-2-methylpropyl)-2-[methyl(5-methyl-6-{[(22)-3-{[2(trimethylsilyl)ethoxy]methyl}-2,3- dihydro-1,3-benzothiazol-2-ylidene]amino}pyridazin-3-yl)amino]1,3-thiazol-4-carboxylate.

To a solution of the product from step A (779 mg, 1.24 mmol, 1 eq.) in methanol (20 ml) cooled to 0°C, sodium borohydride (51.7 mg, 1.37 mmol, 1.1 eq.) was added and the mixture was stirred for 20 minutes. The reaction mixture was partitioned between ethyl acetate and water, and the organic phase was washed with brine, dried (magnesium sulfate) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 24 g RediSep™ silica gel cartridge) eluting with a gradient of 0-50% ethyl acetate in isoheptane gave the title product as a yellow foam (336 mg, 0.53 mmol, 43%).

LC/MS (C29H40N6O4SiS2) 629 [M+H]+; RT 2.74 (LCMS-V-C).

1H NMR (400 MHz, DMSOO δ 7.87-7.83 (m, 1H), 7.66 (d, J=1.1 Hz, 1H), 7.49-7.40 (m, 2H), 7.28-7.21 (m, 1H), 5.85 (s , 2H), 4.62 (t, J=5.2 Hz, 1H), 4.27 (q, J=7.1 Hz, 2H), 3.76 (s, 3H), 3.75-3.66 (m, 3H), 3.42-3.26 (m, 2H), 3.13 (dd, J=14.2, 6.0 Hz, 1H), 2.96 (dd, J=14.2, 8.5 Hz, 1H), 2.45 (d, J=1.0 Hz, 3H), 1.95-1.83 (m, 1H ), 1.31 (t, 3H), 0.95-0.89 (m, 2H), 0.88 (d, 3H), -0.11 (s, 9H).

Step C: ethyl 5-(3-iodo-2-methylpropyl)-2-[methyl(5-methyl-6-{[(22)-3-{[2(trimethylsilyl)ethoxy]methyl}-2,3- dihydro-1,3-benzothiazol-2-ylidene]amino}pyridazin-3-yl)amino]1,3-thiazol-4-carboxylate.

To a solution of the product from step B (469 mg, 0.75 mmol, 1 eq.) in a 6:1 mixture of diethyl ether/acetonitrile (21 ml) was added iodine (284 mg, 1.12 mmol, 1.5 eq.), triphenylphosphine (293 mg, 1.12 mmol, 1.5 eq.) and imidazole (0.07 ml, 1.12 mmol, 1.5 eq.), and the mixture was stirred at ambient temperature for 16 h. The reaction mixture was partitioned between ethyl acetate and 10% aqueous sodium thiosulfate, and the organic phase was dried ( magnesium sulfate) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 12 g RediSep™ silica gel cartridge) eluting with a gradient of 0-30% ethyl acetate in isoheptane gave the title product as a yellow oil (523 mg, 0.71 mmol, 95%).

LC/MS (C ₂₉ H ₃₉ IN ₆ O ₃ SiS ₂ ) 739 [M+H]+; RT 3.23 (LCMS-VC).

1H NMR (400 MHz, DMSOO δ 7.87-7.81 (m, 1H), 7.67 (d, J=1.2 Hz, 1H), 7.52-7.40 (m, 2H), 7.28-7.22 (m, 1H), 5.85 (s , 2H), 4.29 (q, J=7.1 Hz, 2H), 3.78 (s, 3H), 3.75-3.67 (m, 2H), 3.48-3.37 (m, 1H), 3.35-3.27 (m, 1H), 3.17-3.03 (m, 2H), 2.45 (d, J=1.0 Hz, 3H), 1.94-1.78 (m, 1H), 1.32 (t, J=7.1 Hz, 3H), 0.97 (d, J=6.5 Hz , 3H), 0.95-0.87 (m, 2H), -0.13 (s, 9H).

Step D: Ethyl 5-(3-{4-[3-(dimethylamino)prop-1-yn-1-yl]-2-fluorophenoxy}-2-methylpropyl)-2[methyl(5-methyl-6-{ [(22)-3-{ [2-(trimethylsilyl)ethoxy]methyl}-2,3-dihydro-1,3-benzothiazol-2ylidene] amino } pyridazin-3 -yl) amino]-1,3-thiazol- 4-carboxylate.

To a solution of the product from step C (523 mg, 0.71 mmol, 1 eq.) in dimethylformamide (20 ml) was added the product of synthesis 6b (178 mg, 0.92 mmol, 1.3 eq.) and cesium carbonate (692 mg, 2.12 mmol, 3 eq. .), and the mixture was heated at 80°C for 3 hours. The reaction mixture was partitioned between ethyl acetate and brine, and the organic phase was dried (magnesium sulfate) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 24 g RediSep™ silica gel cartridge) eluting with a gradient of 0-5% methanol in dichloromethane gave the title product as a yellow oil (257 mg, 0.32 mmol, 45%).

LC/MS (C ₄₀ H ₅₀ FN ₇ O ₄ SiS ₂ ) 804 [M+H]+; RT 2.66 (LCMS-VC).

1H NMR (400 MHz, DMSOO δ 7.86-7.80 (m, 1H), 7.66 (d, J=1.1 Hz, 1H), 7.49-7.40 (m, 2H), 7.33-7.10 (m, 4H), 5.85 (s , 2H), 4.26 (q, 2H), 4.08-3.95 (m, 2H), 3.77 (s, 3H), 3.75-3.65 (m, 2H), 3.36 (s, 2H), 3.32-3.25 (m, 1H ), 3.17-3.05 (m, 1H), 2.44 (d, J=1.0 Hz, 3H), 2.37-2.24 (m, 1H), 2.18 (s, 6H), 1.29 (t, J=7.1 Hz, 3H) , 1.04 (d, J=6.7 Hz, 3H), 0.96-0.85 (m, 2H), -0.12 (s, 9H).

Step E: ethyl 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)-5-(3-{4[3 -( dimethylamino)prop-1-yn-1-yl]-2-fluorophenoxy}-2-methylpropyl)-1,3-thiazol-4-carboxylate.

To a solution of the product from step D (257 mg, 0.32 mmol, 1 eq.) in tetrahydrofuran (15 ml) was added ethylenediamine (64 μl, 0.96 mmol, 3 eq.) followed by the addition of TBAF (1 M in tetrahydrofuran; 0.96 ml, 0.96 mmol, 3 eq.), and the mixture was heated at 60°C for 18 hours. The reaction mixture was partitioned between ethyl acetate and water, and the organic phase was dried (magnesium sulfate) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 12 g RediSep™ silica gel cartridge) eluting with a gradient of 0-5% methanol in dichloromethane gave the title product as a yellow glass (110 mg, 0.16 mmol, 51%) .

- 184 046336

LC/MS (C ₃₄ H ₃₆ FN ₇ O ₃ S ₂ ) 674 [M+H]+; RT 2.11 (LCMS-VC).

1H NMR (400 MHz, DMSO-de) δ 11.55 (br s, 1H), 7.92 (d, J=8.0 Hz, 1H), 7.69 (s, 1H), 7.54 (d, J=8.0 Hz, 1H), 7.43-7.36 (m, 1H), 7.31 (dd, J=11.9, 1.9 Hz, 1H), 7.27-7.17 (m, 2H), 7.15 (t, J=8.6 Hz, 1H), 4.26 (q, 2H) , 4.08-3.95 (m, 2H), 3.78 (s, 3H), 3.38 (s, 2H), 3.33-3.27 (m, 1H), 3.18-3.07 (m, 1H), 2.47 (d, J=1.0 Hz , 3H), 2.402.27 (m, 1H), 2.20 (s, 6H), 1.31 (t, J=7.1 Hz, 3H), 1.05 (d, J=6.7 Hz, 3H).

Step F: 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)-5-(3-{4-[3(dimethylamino )prop-1-yn-1-yl]-2-fluorophenoxy}-2-methylpropyl)-1,3-thiazol-4-carboxylic acid.

To a solution of the product from step E (110 mg, 0.16 mmol, 1 eq.) in 1,4-dioxane (15 ml) was added lithium hydroxide monohydrate (68.5 mg, 1.63 mmol, 10 eq.) and the mixture was refluxed at for 16 h. The reaction mixture was concentrated in vacuo and the residue was triturated with water (5 ml), filtered, washed with water and dried in vacuo to give the title product as a yellow solid (91.2 mg, 0.14 mmol, 87%) [as salt lithium].

MSHR-ERI (m/z) [M+H] ⁺ , calculated for C ₃₂ H ₃₃ FN ₇ O ₃ S ₂ : 646.2065; found: 646.2091.

Example 117: 2-[[6-(1,3-benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]-(2,3dihydroxypropyl)amino]-5-[3-[2-fluoro-4- [3-(methylamino)prop-1-ynyl]phenoxy]propyl]thiazole-4carboxylic acid

Step A: methyl 5-[3-[4-[3-[tert-butoxycarbonyl(methyl)amino]prop-1-ynyl]-2fluorophenoxy]propyl]-2-[(2,2-dimethyl-1,3- dioxolan-4-yl)methyl-[5-methyl-6-[^)-[3-(2trimethylsilylethoxymethyl)-1,3-benzothiazol-2-ylidene]amino]pyridazin-3-yl]amino]thiazol-4carboxylate.

Using the general Buchwald reaction procedure II, starting from 280 mg of synthesis product 3o (0.47 mmol, 1 eq.) and 193 mg of synthesis product 4a (0.47 mmol, 1 eq.) as the appropriate halide, 400 mg (88%) of the target product were obtained product.

¹ H NMR (500 MHz, DMSO^) δ ppm 7.84 (d, 1H), 7.80 (s, 1H), 7.47 (d, 1H), 7.44 (td, 1H), 7.32 (brd., 1H), 7.26 (td, 1H), 7.22 (d, 1H), 7.16 (t, 1H), 5.87 (s, 2H), 4.55 (m, 2H), 4.53 (m, 1H), 4.20 (br., 2H), 4.15 (t, 2H), 4.07/3.83 (dd+dd , 2H), 3.77 (s, 3H), 3.71 (t, 2H), 3.27 (t, 2H), 2.84 (br., 3H), 2.43 (s, 3H), 2.13 (m, 2H), 1.40 (s , 9H), 1.27/1.20 (s+s, 6H), 0.92 (t, 2H), -0.12 (s, 9H); ¹³ C NMR (125 MHz, DMSO^) δ ppm 129.1, 127.2, 123.2, 123.2, 119.3, 118.9, 115.5, 112.0, 109.2, 74.0, 72.8, 68.4, 66.9, 66.7, 52.0, 49.6, 38.5, 33.8, 30.9, 28.5, 26.6/25.7, 23.1, 17.8, 17.7, -1.0; MSVR-ERI (m/z): [M+H] ⁺ , calculated for C ₄₇ H ₆₁ FN ₇ O ₈ S ₂ Si:

962.3771; found: 962.3767.

Step B: 2-[[6-(1,3-benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]-(2,3dihydroxypropyl)amino]-5-[3-[2-fluoro-4- [3-(methylamino)prop-1-ynyl]phenoxy]propyl]thiazole-4carboxylic acid.

Using a general deprotection and hydrolysis procedure, starting with the product from Step A as the appropriate methyl ester, the desired product was obtained.

MSVR-ERI (m/z): [M+H] ⁺ , calculated for C ₃₂ H ₃₃ FN ₇ O ₅ S ₂ : 678.1963; found: 678.1965.

Example 118: 2-[[6-(1,3-benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]-(3-hydroxy-2methoxypropyl)amino]-5-[3-[2-fluoro- 4-[3-(methylamino)prop-1-ynyl]phenoxy]propyl]thiazole-4carboxylic acid

Step A: methyl 5-[3-[4-[3-[tert-butoxycarbonyl(methyl)amino]prop-1-ynyl]-2fluorophenoxy]propyl]-2-[[3-[tert-butyl(dimethyl)silyl ]oxy-2-methoxypropyl]-[5-methyl-6-[^)-[3-(2trimethylsilylethoxymethyl)-1,3-benzothiazol-2-ylidene]amino]pyridazin-3-yl]amino]thiazol-4

- 185 046336 carboxylate.

Using the general Buchwald reaction procedure II, starting from 400 mg of synthesis product 3q (0.59 mmol, 1 eq.) and 240 mg of synthesis product 4a (0.59 mmol, 1 eq.) as a suitable halide, 534 mg (86%) of the target product were obtained product.

¹ H NMR (500 MHz, DMSO^b) δ ppm 7.84 (dm, 1H), 7.72 (s, 1H), 7.47 (dm, 1H), 7.44 (m, 1H), 7.31 (brd, 1H), 7.25 (m, 1H), 7.21 (dd, 1H), 7.16 (t, 1H), 5.88/5.85 (d+d, 2H), 4.48/4.35 (dd+dd, 2H), 4.20 (brs, 2H), 4.15 (t, 2H), 3.77/3.66 (dd+dd, 2H), 3.76 (m, 1H), 3.76 (s, 3H), 3.71 (m, 2H), 3.27 (m, 2H), 3.23 (s, 3H), 2.84 (brs, 3H), 2.42 (s, 3H ), 2.12 (m, 2H), 1.40 (s, 9H), 0.92 (m, 2H), 0.85 (s, 9H), 0.04 (s, 3H), 0.03 (s, 3H), -0.12 (s, 9H ); ¹³ C NMR (125 MHz, DMSOC) δ ppm 129.1, 127.2, 123.4, 123.2, 119.3, 118.6, 115.5, 112.0, 79.4, 72.8, 68.4, 66.7, 63.1, 58.2, 51.8, 48.4, 38.3, 33.7, 30.9, 2 8.4, 26.2, 23.0, 18.0, 17.7, -1.6 , -5.1, -5.1; MSHR-ERI (m/z): [M+H] ⁺ , calculated for C ₅₁ H ₇₃ FN ₇ O ₈ S ₂ Si ₂ : 1050.4479; found: 1050.4469.

Step B: 2-[[6-(1,3-benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]-(3-hydroxy-2methoxypropyl)amino]-5-[3-[2-fluoro- 4-[3-(methylamino)prop-1-ynyl]phenoxy]propyl]thiazole-4carboxylic acid.

Using a general deprotection and hydrolysis procedure, starting with the product from Step A as the appropriate methyl ester, the desired product was obtained.

MSHR-ERI (m/z): [M+H] ⁺ , calculated for C ₃₃ H ₃₅ FN ₇ O ₅ S ₂ : 692.2120; found: 692.2117.

Example 119: 2-[[6-(1,3-benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]-(4,5dihydroxypentyl)amino]-5-[3 - [2-fluoro-4- [3 -(methylamino)prop-1-ynyl]phenoxy]propyl]thiazole-4carboxylic acid

Using the general deprotection and hydrolysis procedure, starting from the synthesis product 5a, step A, as a suitable methyl ester, the desired product was obtained.

MSHR-ERI (m/z): [M+H] ⁺ , calculated for C ₃₄ H ₃₇ FN ₇ O ₅ S ₂ : 706.2276; found: 706.2274.

Example 120: 2-[[6-(1,3-benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]-(3,4dihydroxybutyl)amino]-5-[3-[2-fluoro-4- [3-[methyl(p-tolylsulfonyl)amino]prop-1ynyl]phenoxy]propyl]thiazole-4-carboxylic acid

Step A: methyl 2-[2-(2,2-dimethyl-1,3-dioxolan-4-yl)ethyl-[5-methyl-6-[(/)-[3-(2trimethylsilylethoxymethyl)-1,3 -benzothiazol-2-ylidene]amino]pyridazin-3-yl]amino]-5-[3-[2-fluoro-4-[3[methyl(p-tolylsulfonyl)amino]prop-1-ynyl]phenoxy]propyl ]thiazole-4-carboxylate.

Using the general Buchwald reaction procedure II, starting with 470 mg of synthesis product 3k (0.71 mmol, 1 eq.) and 290 mg of synthesis product 4a (0.71 mmol, 1 eq.) as the appropriate halide, 660 mg (90%) of the target product were obtained product.

Step B: 2-[[6-(1,3-benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]-(3,4dihydroxybutyl)amino]-5-[3-[2-fluoro-4- [3-[methyl(p-tolylsulfonyl)amino]prop-1ynyl]phenoxy]propyl]thiazole-4-carboxylic acid.

Using a general deprotection and hydrolysis procedure, starting with the product from Step A as the appropriate methyl ester, the desired product was obtained.

- 186 046336

MSHR-ERI (m/z): [M+H]+, calculated for C40H41FN7O7S3: 846.2208; found: 846.2201.

Example 121: 2-[[6-(1,3-benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]-(3,4dihydroxybutyl)amino]-5-[3-[2-fluoro-4- [3-[(4-methoxyphenyl)methylmethylamino]prop-1 inyl]phenoxy]propyl]thiazole-4-carboxylic acid

Step A: methyl 2-[2-(2,2-dimethyl-1,3-dioxolan-4-yl)ethyl-[5-methyl-6-[^)-[3-(2trimethylsilylethoxymethyl)-1,3- benzothiazol-2-ylidene]amino]pyridazin-3-yl]amino]-5-[3-[2-fluoro-4-[3[(4-methoxyphenyl)methylmethylamino]prop-1-ynyl]phenoxy]propyl]thiazole -4-carboxylate.

Using the general Buchwald reaction procedure II, starting with 625 mg of synthesis 3j (1 mmol, 1 eq.) and 407 mg of synthesis 4a (1 mmol, 1 eq.) as the appropriate halide, 550 mg (55%) of the desired product.

Step B: 2-[[6-(1,3-benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]-(3,4dihydroxybutyl)amino]-5-[3-[2-fluoro-4- [3-[(4-methoxyphenyl)methylmethylamino]prop-1 inyl]phenoxy]propyl]thiazole-4-carboxylic acid.

Using a general deprotection and hydrolysis procedure, starting with the product from Step A as the appropriate methyl ester, the desired product was obtained.

MSHR-ERI (m/z): [M+H] ⁺ , calculated for C ₄₁ H ₄₃ FN ₇ O ₆ S ₂ : 812.2695; found: 812.2674.

Example 122: 2-[[6-(1,3-benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]-(2,3dihydroxypropyl)amino]-5-[3-[4-[3-( dimethylamino)prop-1-ynyl]-2-fluorophenoxy]propyl]thiazole-4carboxylic acid

Step A: methyl 5-[3-[4-[3-(dimethylamino)prop-1-ynyl]-2-fluorophenoxy]propyl]-2-[(2,2-dimethyl1,3-dioxolan-4-yl) methyl-[5-methyl-6-[^)-[3-(2-trimethylsilylethoxymethyl)-1,3-benzothiazol-2ylidene]amino]pyridazin-3-yl]amino]thiazol-4-carboxylate.

Using the general Buchwald reaction procedure II, starting from 380 mg of synthesis product 3p (0.75 mmol, 1 eq.) and 306 mg of synthesis product 4a (0.75 mmol, 1 eq.) as the appropriate halide, 480 mg (72%) of the target product were obtained product.

1H NMR (500 MHz, DMSO-d,) δ ppm 7.84 (dm, 1H), 7.78 (s, 1H), 7.47 (dm, 1H), 7.44 (m, 1H), 7.30 (dd, 1H), 7.26 (m, 1H), 7.21 (m, 1H), 7.15 (t, 1H), 5.86 (s, 2H), 4.57/4.53 (m+m, 2H), 4.52 (m, 1H), 4.15 (t, 2H), 4.07/3.83 (dd+dd, 2H), 3.77 (s, 3H), 3.71 (m, 2H), 3.38 (s, 2H), 3.27 (m, 2H), 2.43 (s, 3H), 2.20 (s, 6H), 2.13 (m, 2H), 1.27 ( s, 3H), 1.20 (s, 3H), 0.92 (m, 2H), -0.12 (s, 9H); ¹³ C NMR (125 MHz, DMSO-d,) δ ppm 163.1, 157.6, 156.8/151.7, 141.5, 137.6, 134.9, 128.9, 127.1, 123.4, 123.2, 119.2, 118.8, 115.4, 111.9, 85.1, 84.1, 73.9, 7 2.7, 68.3, 66.9, 66.6, 51.9, 49.6, 48.1, 44.2, 30.9, 26.6, 25.6, 23.0, 17.8, 17.7, -1.0; MSHR-ERI (m/z): [M+H] ⁺ , calculated for C ₄₃ H ₅₅ FN ₇ O ₆ S ₂ Si: 876.3403; found: 876.3408.

Step B: 2-[[6-(1,3-benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]-(2,3dihydroxypropyl)amino]-5-[3-[4-[3-( dimethylamino)prop-1-ynyl]-2-fluorophenoxy]propyl]thiazole-4carboxylic acid.

Using a general deprotection and hydrolysis procedure, starting with the product from Step A as the appropriate methyl ester, the desired product was obtained.

MSHR-ERI (m/z): [M+H] ⁺ , calculated for C ₃₃ H ₃₅ FN ₇ O ₅ S ₂ : 692.2120; found: 692.2117.

Example 123: 2-[[6-(1,3-Benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]-(3,4dihydroxybutyl)amino]-5-[3-[4-[3-( dimethylamino)prop-1-ynyl]-2-fluorophenoxy]propyl]thiazol-4- 187 046336 carboxylic acid

Step A: methyl 5-[3-[4-[3-(dimethylamino)prop-1-ynyl]-2-fluorophenoxy]propyl]-2-[2-(2,2-dimethyl1,3-dioxolane-4- yl)ethyl-[5-methyl-6-[^)-[3-(2-trimethylsilylethoxymethyl)-1,3-benzothiazol-2ylidene] amino]pyridazin-3 -yl] amino]thiazol-4-carboxylate.

Using the general Buchwald reaction procedure II, starting from 350 mg of synthesis product 3i (0.67 mmol, 1 eq.) and 275 mg of synthesis product 4a (0.67 mmol, 1 eq.) as a suitable halide, 510 mg (85%) of the target product were obtained product.

Step B: 2-[[6-(1,3-benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]-(3,4dihydroxybutyl)amino]-5-[3-[4-[3-( dimethylamino)prop-1-ynyl]-2-fluorophenoxy]propyl]thiazole-4carboxylic acid.

Using a general deprotection and hydrolysis procedure, starting with the product from Step A as the appropriate methyl ester, the desired product was obtained.

MSHR-ERI (m/z): [M+H]+, calculated for C ₃₄ H ₃₇ FN ₇ O ₅ S ₂ : 706.2276; found: 706.2270.

Example 124: 2-[[6-(1,3-Benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]-[5-[2(dimethylamino)ethylmethylamino]-4-hydroxypentyl]amino]-5- [3-[4-[3-(dimethylamino)prop-1-ynyl]-2fluorophenoxy]propyl]thiazole-4-carboxylic acid

Step A: methyl 2-[[4-[tert-butyl(diphenyl)silyl]oxy-5-[2-(dimethylamino)ethylmethylamino]pentyl]-[5-methyl-6-[^)-[3-(2 -trimethylsilylethoxymethyl)-1,3-benzothiazol-2-ylidene]amino]pyridazin3 -yl]amino]-5-[3-[4-[3-(dimethylamino)prop-1-ynyl]-2-fluorophenoxy]propyl] thiazole-4-carboxylate.

Using the general tosylate alkylation procedure from synthesis 5b and N,N',N'trimethylethane-1,2-diamine as a suitable amine, the desired product was obtained.

Step B: 2-[[6-(1,3-benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]-[5-[2-(dimethylamino)ethylmethylamino]-4-hydroxypentyl]amino]-5 -[3-[4-[3-(dimethylamino)prop-1-ynyl]-2fluorophenoxy]propyl]thiazole-4-carboxylic acid.

Using a general deprotection and hydrolysis procedure, starting with the product from Step A as the appropriate methyl ester, the desired product was obtained.

MSVR-ERI (m/z): [M+H] ⁺ , calculated for C ₄₀ H ₅₁ FN9O ₄ S ₂ : 804.3484; found: 804.3487.

- 188 046336

Example 125: 2-[[6-(1,3-Benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]-(5-hydroxypentyl)amino]thiazole-4-carboxylic acid

Step A: methyl 2-[tert-butoxycarbonyl-[5-[tert-butyl(dimethyl)silyl]oxypentyl]amino]thiazole 4-carboxylate.

Using the general Mitsunobu reaction procedure, starting with methyl 2-(tert-butoxycarbonylamino)thiazole-4-carboxylate and 5-[tert-butyl(dimethyl)silyl]oxypentan-1-ol as starting materials and carrying out the reaction in THF at RT . within 2 hours, 565 mg (82%) of the target product was obtained.

¹ H NMR (400 MHz, DMSO^) δ ppm 8.08 (s, 1H), 4.05 (t, 2H), 3.79 (s, 3H), 3.56 (t, 2H), 1.65 (m, 2H), 1.52 (s, 9H), 1.48 (m, 2H), 1.31 (m, 2H), 0.83 (s, 9H), -0.02 (s, 6H).

Step B: Methyl 2-[5-[tert-butyl(dimethyl)silyl]oxypentylamino]thiazole-4-carboxylate.

Using the general HFIP deprotection procedure, starting with the product from Step A as the appropriate carbamate, 484 mg (113%) of the desired product was obtained.

¹ H NMR (400 MHz, DMSO^) δ ppm 7.78 (t, 1H), 7.50 (s, 1H), 3.57 (t, 2H), 3.73 (s, 3H), 3.20 (q, 2H), 1.59-1.42 (m, 4H), 1.40-1.29 (m, 2H), 0.85 (s, 9H), 0.02 (s, 6H).

Step C: methyl 2-[5-[tert-butyl(dimethyl)silyl]oxypentyl-[5-methyl-6-[^)-[3-(2trimethylsilylethoxymethyl)-1,3-benzothiazol-2-ylidene]amino] pyridazin-3-yl]amino]thiazol-4carboxylate.

Using the general Buchwald reaction procedure II, starting from the product from step B and the product of synthesis 4a as a suitable halide, 535 mg (61%) of the desired product were obtained.

¹ H NMR (400 MHz, DMSO^) δ ppm 7.97 (s, 1H), 7.84 (d, 1H), 7.70 (s, 1H), 7.46 (t, 1H), 7.43 (t, 1H), 7.25 (dt, 1H), 5.86 (s, 2H), 4.45 (t, 2H), 3.81 (s, 3H), 3.72 (t, 2H), 3.55 (t, 2H), 2.46 (s, 3H), 1.72 (m, 2H), 1.50 (m, 2H), 1.38 ( m, 2H), 0.92 (t, 2H), 0.79 (s, 9H), -0.04 (s, 6H), -0.11 (s, 9H).

Step D: 2-[[6-(1,3-benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]-(5-hydroxypentyl)amino]thiazol-4-carboxylic acid.

Using the general deprotection and hydrolysis procedure followed by repurification by reverse phase preparative chromatography (C18, 25 mM NH4HCO3 water:MeCN), the desired product was obtained from the product from step C.

MSHR-ERI (m/z): [M+H]+, calculated for C ₂₁ H ₂₃ N ₆ O ₃ S ₂ : 471.1273; found: 471.1271.

Example 126: 2-[[6-(1,3-benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]-(3hydroxypropyl)amino]-5-[3-[4-[3-(dimethylamino) prop-1-ynyl]-2-fluorophenoxy]propyl]thiazole-4carboxylic acid

\

Stage A: ethyl 3-bromo-6-chloro-2-oxohexanoate.

To 2 g of ethyl 6-chloro-2-oxohexanoate (10.4 mmol, 1 eq.) in 20 ml of CCl ₄ , 0.6 ml of bromine (11.5 mmol, 1.1 eq.) was added dropwise, and the mixture was stirred at room temperature. for 1 hour. The mixture was then poured into 50 ml of 1 M aqueous Na ₂ S ₂ O ₃ and extracted with 2x50 ml ethyl acetate, and the combined organic layers were washed with brine, dried, concentrated and purified by silica gel chromatography using petroleum ether and EtOAc as eluents to obtain 2.05 g (73%) of the target product.

- 189 046336 ¹ H NMR (400 MHz, DMSO-dg) δ ppm 5.25 (dd, 1H), 4.30 (q, 2H), 3.71 (t, 2H), 2.21-1.80 (m, 4H), 1.29 (t, 3H); GC/MS-EI: [M] ⁺ = 270.

Step B: 3-[tert-butyl(dimethyl)silyl]oxypropan-1-amine.

To 8.25 g 3-aminopropan-1-ol (110 mmol, 1 eq.), 23 ml N,N-diethylethanamine (170 mmol, 1.5 eq.) and 30 mg imidazole (0.44 mmol, 0.004 eq.) in 200 ml DCM 18.2 g of tert-butyldimethylsilyl chloride (150 mmol, 1.4 eq.) were added and the mixture was stirred at room temperature. for 15 hours. The reaction mixture was then poured into 250 ml water and extracted with 2x200 ml DCM, the combined organic layers were washed with 50 ml water and 50 ml brine, dried and concentrated to obtain 20.9 g (100%) of the expected product.

1H NMR (400 MHz, DMSO-dg) δ ppm 3.60 (t, 2H), 2.55 (t, 2H), 1.50 (quint, 2H), 0.80 (s, 9H), 0.10 (s, 6H); IR: 3000-2900, 1100, 830-770.

Step C: 3-[tert-butyl(dimethyl)silyl]oxypropylthiourea.

To 5.3 g of 9-fluorenylmethoxycarbonyl isothiocyanate (18.9 mmol, 1 eq.) and 6.4 ml of DIPEA (37.6 mmol, 2 eq.) in 50 ml of DCM, a solution of 3.5 g of the product from step B (18.5 mmol, 1 eq.) in 10 ml DCM, and the mixture was stirred at room temperature. for 2 hours. After concentration, the crude product was purified by silica gel chromatography using petroleum ether and EtOAc as eluents to obtain 3.87 g (80%) of the expected product.

1H NMR (400 MHz, DMSOD ₆ ) δ ppm 7.49 (brs, 1H), 6.83 (brs, 1H), 7.21 (s, 1H), 3.57 (t, 2H), 3.34/3.02 (m, 2H), 1.61 (m, 2H), 0.83 (s, 9 H ), 0.02 (s, 6 N); IR: 3275, 1619.

Step D: Ethyl 2-[3-[tert-butyl(dimethyl)silyl]oxypropylamino]-5-(3-chloropropyl)thiazole-4carboxylate.

A solution of 7.4 g of the product from step C (29.7 mmol, 1 eq.), 8.1 g of the product from step A (30 mmol, 1 eq.) and 8.2 ml of N,N-diethylethanamine (59.4 mmol, 2 eq.) in 206 ml of ethanol Heated at reflux for 18 hours. After concentration, the crude residue was taken up in DCM, washed with water, dried, concentrated and purified by silica gel chromatography using petroleum ether and EtOAc as eluents to obtain 10.1 g (81%) of the expected product .

1H NMR (400 MHz, DMSOD ₆ ) δ ppm 7.56 (t, 1H), 4.17 (q, 2H), 3.62 (2t, 4H), 3.18 (q, 2H), 3.06 (t, 2H), 1.95 (m, 2H), 1.69 (m, 2H), 1.23 (t, 3H), 0.83 (s, 9H), 0.01 (s, 6H); IR: 3197, 1716, 1586.

Step E: ethyl 2-[3-[tert-butyl(dimethyl)silyl]oxypropyl-(6-chloropyridazin-3-yl)amino]-5-(3chloropropyl)thiazole-4-carboxylate.

To 10 g of the product from step D (23.7 mmol, 1 eq.) and 5.8 g of 3,6-dichloro-4-methylpyridazine (35.6 mmol, 1.5 eq.) in 120 ml of THF, 2.1 g of sodium hydride (52.1 mmol, 2.2 eq.) at 50°C. The cooled mixture was poured onto ice and extracted with 2x200 ml ethyl acetate. The combined organic layers were washed with brine, dried, concentrated and purified by silica gel chromatography using petroleum ether and EtOAc as eluents to give 4.3 g (32%) of the expected product.

1H NMR (400 MHz, DMSOC6) δ ppm 7.76 (s, 1H), 4.39 (t, 2H), 4.26 (q, 2H), 3.68 (m, 4H), 3.19 (t, 2H), 2.39 (s, 3H), 2.06 (m, 2H), 1.89 (m, 2H), 1.29 (t, 3H), 0.83 (s, 9H), 0.01 (s, 6H); IR: 1704, 1592, 1195, 1095 + 1066, 649.

Step F: Ethyl 2-[3-[tert-butyl(dimethyl)silyl]oxypropyl-(6-chloro-5-methylpyridazin-3yl)amino]-5-(3-iodopropyl)thiazole-4-carboxylate.

A mixture of 4.6 g of the product from step E (8.42 mmol, 1 eq.) and 12.6 g of sodium iodide (84.2 mmol, 10 eq.) in 58 ml of acetone was kept at 60°C for 18 hours. After concentration, the residue was added to ethyl acetate and washed water and brine, dried and concentrated to obtain the desired product, which was used in the next step without further purification.

1H NMR (400 MHz, DMSOD ₆ ) δ ppm 7.78 (s, 1H), 4.41 (t, 2H), 4.29 (q, 2H), 3.71 (t, 2H), 3.31 (t, 2H), 3.17 (t, 2H), 2.41 (s, 3H), 2.12 (m, 2H), 1.91 (m, 2H), 1.32 (t, 3H), 0.84 (s, 9H), 0.02 (s, 6H).

Step G: ethyl 2-[3-[tert-butyl(dimethyl)silyl]oxypropyl-(6-chloro-5-methylpyridazin-3yl)amino]-5-[3-[4-[3-(dimethylamino)prop- 1-ynyl]-2-fluorophenoxy]propyl]thiazole-4-carboxylate.

A mixture of 800 mg of the product from step F (1.25 mmol, 1 eq.), 821 mg of cesium carbonate (2.5 mmol, 2 eq.) and 290 mg of the product of synthesis 6b (1.5 mmol, 1.2 eq.) in 6 ml of MeCN was heated to reflux for 2 hours. After diluting the mixture with 100 ml ethyl acetate, the organic layer was washed with 50 ml water and 50 ml brine, dried, concentrated and purified by silica gel chromatography using DCM and MeOH (containing 7N NH ₃ ) as eluents to obtain 500 mg (57%) of the target product.

1H NMR (400 MHz, DMSOD ₆ ) δ ppm 7.77 (m, 1H), 7.27 (dd, 1H), 7.18 (d, 1H), 7.11 (t, 1H), 4.40 (t, 2H), 4.23 (q, 2H), 4.11 (t, 2H), 3.69 (t, 2H), 3.40 (s, 2H), 3.24 (t, 2H), 2.40 (s, 3H), 2.21 (s, 6H), 2.10 (m, 2H), 1.89 (m, 2H), 1.27 ( t, 3H), 0.83 (s, 9H), 0.00 (s, 6H); IR: 3600-3100, 2237, 1716, 835, 777.

Step H: ethyl 2-[[6-(1,3-benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]-(3hydroxypropyl)amino]-5-[3-[4-[3-(dimethylamino )prop-1-ynyl]-2-fluorophenoxy]propyl]thiazole-4carboxylate.

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Mixture of 100 mg product from step G (0.142 mmol, 1 eq.), 28 mg 1,3-benzothiazol-2-amine (0.185 mmol, 1.3 eq.), 10 mg 4,5-bis(diphenylphosphino)-9,9 -dimethylxanthene (0.017 mmol, 0.12 equiv.), 8 mg tris(dibenzylideneacetone)dipalladium(0) (8.52 µmol, 0.06 equiv.) and 72 µl DIPEA (0.426 mmol, 3 equiv.) in 0.7 ml X-methyl-2- pyrrolidone was kept at 120°C for 18 hours. The mixture was then poured into 50 ml of DCM, the organic layer was washed with 10 ml of water and 10 ml of saturated aqueous lithium chloride, dried, concentrated and purified by silica gel chromatography using DCM and MeOH ( containing 7 N NH ₃ ) as eluents to obtain 40 mg (40%) of the target product.

1H NMR (400 MHz, DMSO-de) δ ppm 11.54 (brs, 1H), 7.91 (d, 1H), 7.69 (s, 1H), 7.53 (d, 1H), 7.39 (t, 1H), 7.30 (dd, 1H), 7.22 (dd, 1H), 7.21 (dd, 1H), 7.16 (t, 1H), 4.72 (t, 1H), 4.41 (t, 2H), 4.25 (q, 2H), 4.16 (t, 2H), 3.48 (q, 2H), 3.39 ( s, 2H), 3.27 (t, 2H), 2.46 (s, 3H), 2.2 (s, 6H), 2.13 (m, 2H), 1.87 (m, 2H), 1.29 (t, 3H); IR: 3414, 3400-2200, 1709, 1602, 1149/1087.

Step I: 2-[[6-(1,3-benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]-(3-hydroxypropyl)amino]-5[3-[4-[3 -(dimethylamino )prop-1-ynyl]-2-fluorophenoxy]propyl]thiazole-4-carboxylic acid.

A mixture of the product from step H and LiOHxH ₂ O (4 eq.) in 1,4-dioxane and water was stirred at room temperature. for 3 hours. After adding an additional 1 eq. LiOHxH ₂ O in water, the reaction mixture was stirred at room temperature. for 18 hours. After slowly adding 2N. HCl solution, the reaction mixture was concentrated and purified using reverse phase preparative chromatography (0.2 M NH4CO3 in water:MeCN) to obtain the target product.

MSHR-ERI (m/z): [M+H]+, calculated for C ₃₃ H ₃₅ FN ₇ O ₄ S ₂ : 676.2170; found: 676.2169.

Example 127: 2-[[6-(1,3-benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]-(3hydroxypropyl)amino]-5-[3-[2-fluoro-4-[3 -(methylamino)prop-1-ynyl]phenoxy]propyl]thiazole-4carboxylic acid

Step A: methyl 5-[3-[4-[3-[tert-butoxycarbonyl(methyl)amino]prop-1-ynyl]-2fluorophenoxy]propyl]-2-[3-[tert-butyl(dimethyl)silyl] hydroxypropyl-[5-methyl-6-[(7)-[3-(2trimethylsilylethoxymethyl)-1,3-benzothiazol-2-ylidene]amino]pyridazin-3-yl]amino]thiazol-4carboxylate.

Using the general Buchwald reaction procedure II, starting from 300 mg of synthesis product 3n (0.46 mmol, 1 eq.) and 187 mg of synthesis product 4a (0.46 mmol, 1 eq.) as a suitable halide, 395 mg (83%) of the target product were obtained product.

¹ H NMR (500 MHz, DMSO-dg) δ ppm 7.82 (dd, 1H), 7.60 (s, 1H), 7.44 (m, 1H), 7.44 (dd, 1H), 7.31 (dd, 1H), 7.24 (m, 1H), 7.20 (m, 1H), 7.15 (t, 1H), 5.84 (s, 2H), 4.39 (t, 2H), 4.20 (s, 2H), 4.14 (t, 2H), 3.76 (s, 3H), 3.70 (t, 2H), 3.70 ( t, 2H), 3.25 (t, 2H), 2.84 (s, 3H), 2.42 (s, 3H), 2.11 (m, 2H), 1.91 (m, 2H), 1.40 (s, 9H), 0.91 (t , 2H), 0.85 (s, 9H), 0.01 (s, 6H), -0.12 (s, 9H); ¹³ C NMR (125 MHz, DMSO-de) δ ppm 162.2, 147.5, 137.6, 129.1, 127.2, 123.4, 123.2, 119.3, 117.5, 115.4, 112.0, 79.7, 72.8, 68.4, 66.7, 60.5, 51.9, 44.6, 38.1 , 33.8, 30.9, 30.4, 28.6, 26.3, 23.1, 17.9, 17.8, -0.9, -5.0; MSHR-ERI (m/z): [M+H]+, calculated for C ₅₀ H ₇₁ FN ₇ O ₇ S ₂ Si ₂ : 1020.4373; found: 1020.4365.

Step B: 2-[[6-(1,3-benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]-(3-hydroxypropyl)amino]-5[3-[2-fluoro-4-[ 3-(methylamino)prop-1-ynyl]phenoxy]propyl]thiazole-4-carboxylic acid.

Using a general deprotection and hydrolysis procedure, starting with the product from Step A as the appropriate methyl ester, the desired product was obtained.

MSVR-ERI (m/z): [M+H]+, calculated for C ₃₂ H ₃₃ FN ₇ O ₄ S ₂ : 662.2014; found: 662.2016.

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Example 128: 2-[[6-(1,3-benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]-(5-hydroxypentyl)amino]5-[3-[2-fluoro-4-[ 3-(methylamino)prop-1-ynyl]phenoxy]propyl]thiazole-4-carboxylic acid

ABOUT

Using the general deprotection and hydrolysis procedure starting from the appropriate methyl ester from Synthesis 5e, Step A, the desired product was obtained.

MSHR-ERI (m/z): [M+H]+, calculated for C ₃₄ H ₃ 7FN7O ₄ S ₂ : 690.2327; found: 690.2318.

Example 130: 3-[3-[4-[3-[2-[[6-(1,3-benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]-(4hydroxybutyl)amino]-4- carboxythiazol-5-yl]propoxy]-3-fluorophenyl]prop-2ynyldimethylammonio]propane-1-sulfonate.

Step A: 3-[3-[4-[3-[2-[4-[tert-butyl(dimethyl)silyl]oxybutyl-[5-methyl-6-[^)-[3-(2trimethylsilylethoxymethyl)-1 ,3-benzothiazol-2-ylidene]amino]pyridazin-3-yl]amino]-4methoxycarbonylthiazol-5-yl]propoxy]-3-fluorophenyl]prop-2-ynyldimethylammonio]propane-1sulfonate.

To a suspension of 660 mg of synthesis product 5d, stage A (0.7 mmol, 1 eq.), in 5 ml of dry acetonitrile, 257 mg of oxathiolane 2,2-dioxide (2.1 mmol, 3 eq.) was added in one portion. The resulting mixture was stirred overnight at room temperature. The crude product was purified by flash column chromatography to obtain 550 mg (75%) of the title product.

¹ HNMR (500 MHz, DMSO-06) δ ppm 7.83-7.20 (m, 4h), 7.62 (s, 1H), 7.58 (dd, 1H), 7.41 (m, 1H), 7.22 (t, 1H), 5.83 (s, 2H), 4.55 (s, 2H) , 4.40 (t, 2H), 4.17 (t, 2H), 3.77 (s, 3H), 3.70 (t, 2H), 3.60 (t, 2H), 3.57 (t, 2H), 3.25 (t, 2H), 3.12 (s, 6H), 2.52 (t, 2H), 2.42 (s, 3H), 2.12 (m, 2H), 2.12 (m, 2H), 2.05 (m, 2H), 1.73 (m, 2H), 0.90 (t, 2H), 0.78 (s, 9H), -0.03 (s, 6H), -0.13 (s, 9H); ¹³ C NMR (125 MHz, DMSO-06) δ ppm 163.2, 148.5, 137.5, 129.8, 120.0, 117.5, 115.3, 89.6, 72.8, 68.5, 66.7, 63.1, 62.5, 54.6, 51.9, 50.3, 48.2, 46.5, 31.0, 29 .7, 26.2, 23.7, 23.2, 19.6, 17.9, 17.7, -1.0, -4.9; HRMS-ERI (m/z): [M+2H] ²⁺ , calculated for C50H7 ₄ FN7O8S ₃ Si2: 535.7136; found: 535.7136.

Step B: 3-[3-[4-[3-[2-[[6-(1,3-benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]-(4hydroxybutyl)amino]-4- carboxythiazol-5-yl]propoxy]-3-fluorophenyl]prop-2ynyldimethylammonio]propane-1-sulfonate.

Using a general deprotection and hydrolysis procedure, starting with the product from Step A as the appropriate methyl ester, the desired product was obtained.

MSHR-ERI (m/z): [M+H]+, calculated for C ₃ 7H ₄₃ FN7O ₇ S ₃ : 812.2365; found: 812.2357.

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Example 131: 2-[[6-(1,3-Benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]-(4-hydroxy-5morpholinopentyl)amino]thiazol-4-carboxylic acid

Step A: Methyl 2-[tert-butoxycarbonyl-[4-[tert-butyl(dimethyl)silyl]oxy-5morpholinopentyl]amino]thiazole-4-carboxylate.

Using the general Mitsunobu reaction procedure, starting with 639 mg of methyl 2-(tert-butoxycarbonylamino)thiazole-4-carboxylate (2.47 mmol, 1.5 eq.) and 500 mg of synthesis product 2e (1.65 mmol, 1 eq.) as the appropriate alcohol, we obtained 730 mg (81%) of the target product.

1H NMR (400 MHz, DMSO^) δ ppm 8.08 (s, 1H), 4.09-4.03 (m, 2H), 3.79 (s+m, 4H), 3.51 (t, 4H), 2.392.28 (m, 4H), 2.21 (ddd, 2H). 1.85-1.73 (m, 1H), 1.70-1.59 (m, 1H), 1.53 (s+m, 10H), 1.39-1.28 (m, 1H), 0.79 (s, 9H), 0.02 (s, 3H), -0.01 (s, 3H); ¹³ C NMR (100 MHz, DMSO^) 160.3, 140.6, 124.2, 71.2, 68.9, 66.2, 64.7, 59.8, 54.2, 51.8, 46.8, 32.3, 27.6, 25.7, 23.2, 21.8, 21.5 , 20.8, 17.7, 14.1, -4.3, -4.9; LC-MS-ESI (m/z): [M+H] ⁺ , calculated for C ₂ 5H ₄₆ N ₃ O ₆ SSi: 544.3; found: 544.3.

Step B: methyl 2-[[4-[tert-butyl(dimethyl)silyl]oxy-5-morpholinopentyl]amino]thiazole-4carboxylate.

Using the general HFIP deprotection procedure, starting from 304 mg of the product from Step A (0.56 mmol) as the appropriate carbamate, 231 mg (93%) of the desired product was obtained.

1H NMR (400 MHz, DMSOM6) δ ppm 7.81 (t, 1H), 7.49 (s, 1H), 3.79 (brs, 1H), 3.73 (s, 3H), 3.53 (t, 4H), 3.21 (q, 2H), 2.42-2.23 (m, 4H) , 2.23 (ddd, 2H), 1.69-1.34 (m, 4H), 0.84 (s, 9H), 0.05 (s, 3H), 0.03 (s, 3H); ¹³ C NMR (100 MHz, DMSOM6) 168.3, 161.6, 142.2, 116.3, 66.2, 65.2, 59.8, 54.3, 51.5, 44.5, 32.7, 25.8, 24.5, 17.8, -4.2, -4.8; LC-MS-ESI (m/z): [M+H] ⁺ , calculated for C ₂ 0H ₃ 8N ₃ O4SSi: 444.2; found: 444.3.

Step C: methyl 2-[[4-[tert-butyl(dimethyl)silyl]oxy-5-morpholinopentyl]-[5-methyl-6-[^)-[3-(2trimethylsilylethoxymethyl)-1,3-benzothiazol- 2-ylidene]amino]pyridazin-3-yl]amino]thiazol-4carboxylate.

Using the general Buchwald reaction procedure II, starting from 231 mg of the product from step B (0.52 mmol, 1 eq.) and 315 mg of the product of synthesis 4a (0.77 mmol, 1.5 eq.) as the appropriate halide, 344 mg (81%) were obtained target product.

1H NMR (500 MHz, DMSOM6) δ ppm 7.97 (s, 1H), 7.85-7.23 (4H), 7.69 (s, 1H), 5.86 (d, 2H), 4.49 (t, 2H), 3.81 (m, 1H), 3.81 (s, 3H), 3.71 (t, 2H), 3.50-2.13 (8H), 2.45 (s, 3H), 2.20 (m, 2H), 1.83-1.73 (m+m, 2H), 1.561.41 (m+m, 2H), 0.91 (t, 2H), 0.75 (s, 9H), 0.01-(-0.05) (s+s, 6H), -0.12 (s, 9H); ¹³ C NMR (125 MHz, DMSOd6) δ ppm 162.3, 160.8, 157.6, 155.3, 123.1, 117.9, 72.9, 69.0, 66.7, 65.4, 52.3, 47.1, 32.4, 26.2, 22.9, 17.9, 17.8, -1.0, -3.8, -4.5; MSHR-ERI (m/z): [M+H] ⁺ , calculated for C38H60N ₇ O ₅ S ₂ Si ₂ : 814.3630; found: 814.3629.

Step D: 2-[[6-(1,3-benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]-(4-hydroxy-5morpholinopentyl)amino]thiazol-4-carboxylic acid.

Using the general deprotection and hydrolysis procedure followed by purification by reverse phase preparative chromatography (C18, 25 mM NH4HCO3 in water:MeCN), the desired product was obtained from the product from step C.

MSHR-ERI (m/z): [M+H] ⁺ , calculated for C ₂₅ H ₃₀ N ₇ O ₄ S ₂ : 556.1795; found: 556.1807.

Example 132: 2-[[6-(1,3-Benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]-(4-hydroxy-5 methoxypentyl)amino]thiazol-4-carboxylic acid

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Step A: Methyl 2-[tert-butoxycarbonyl-(5-methoxy-4triisopropylsilyloxypentyl)amino]thiazole-4-carboxylate.

Using the general Mitsunobu reaction procedure, starting from 284 mg of methyl 2-(tert-butoxycarbonylamino)thiazole-4-carboxylate (1.10 mmol, 1.1 eq.) and 291 mg of synthesis product 2c (1.00 mmol, 1 eq.) as the appropriate alcohol, we obtained 456 mg (86%) of the target product.

1H NMR (400 MHz, DMSO-06) δ ppm 8.08 (s, 1H), 4.07 (dt, 2H), 3.89 (qv, 1H), 3.79 (s 3H), 3.27 (dd, 2H), 3.23 (s, 3H), 1.73 (qv, 2H), 1.52 ( s, 9H), 1.50-1.38 (m, 2H), 0.96 (s, 21H); ¹³ C NMR (100 MHz, DMSO-de) δ ppm 161.4, 160.3, 140.6, 124.2, 75.7, 70.5, 58.4, 51.9, 46.8, 31.2, 22.5, 17.89, 17.87, 11.9; LC-MS-ESI (m/z): [M+H]+, calculated for C ₂₅ H ₄₇ N ₂ O ₆ SSi: 531.3; found: 531.4.

Step B: Methyl 2-[(5-methoxy-4-triisopropylsilyloxypentyl)amino]thiazole-4-carboxylate.

Using the general HFIP deprotection procedure, starting from 444 mg of the product from Step A (0.84 mmol) as the appropriate carbamate, 334 mg (93%) of the desired product was obtained.

1H NMR (500 MHz, DMSO^) δ ppm 7.82 (t, 1H), 7.50 (s, 1H), 3.91 (t, 1H), 3.73 (s, 3H), 3.28 (d, 2H), 3.24 (s, 3H), 3.21 (q, 2H), 1.61 (m, 2H), 1.52 (m, 2H), 1.01 (m, 21H); ¹³ C NMR (125 MHz, DMSO^) δ ppm 162.0, 116.9, 76.7, 71.2, 58.9, 52.0, 45.0, 32.2, 24.4; MSHR-ERI (m/z): [M+H] ⁺ , calculated for C20H ₃₉ N2O4SSi: 431.2394; found: 431.2392.

Step C: methyl 2-[(5-methoxy-4-triisopropylsilyloxypentyl)-[5-methyl-6-[^)-[3-(2trimethylsilylethoxymethyl)-1,3-benzothiazol-2-ylidene]amino]pyridazin-3 -yl]amino]thiazole-4carboxylate.

Using the general Buchwald reaction procedure II, starting from 324 mg of the product from step B (0.75 mmol, 1 eq.) and 337 mg of the product of synthesis 4a (0.83 mmol, 1.1 eq.) as the appropriate halide, 540 mg (90%) were obtained target product.

1H NMR (400 MHz, DMSO-do) δ ppm 7.97 (s, 1H), 7.85-7.22 (4H), 7.68 (s, 1H), 5.86 (s, 2H), 4.48 (m, 2H), 3.88 (m, 1H), 3.81 (s, 3H), 3.71 (t, 2H), 3.26 (d, 2H), 3.21 (s, 3H), 2.44 (s, 3H), 1.79 (m, 2H), 1.53 (m, 2H), 0.91 (t, 2H), 0.91 ( m, 21H), -0.13 (s, 9H); ¹³ C NMR (125 MHz, DMSO^) δ ppm 162.3, 160.7, 123.1, 117.7, 76.4, 73.0, 71.0, 66.7, 58.8, 52.2, 47.2, 31.4, 22.5, 18.3, 12.3, 17.8, 17.8, -1.0 MSVR-ERI (m/z): [M +H] ⁺ , calculated for C ₃₈ H61N ₆ O ₅ S ₂ Si ₂ : 801.3678; found: 801.3671.

Step D: 2-[[6-(1,3-benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]-(4-hydroxy-5methoxypentyl)amino]thiazol-4-carboxylic acid.

Using the general deprotection and hydrolysis procedure followed by purification by reverse phase preparative chromatography (C18, 25 mM NH4HCO3 in water:MeCN), the desired product was obtained from the product from step C.

MSHR-ERI (m/z): [M+H] ⁺ , calculated for C ₂₂ H ₂₅ N ₆ O ₄ S ₂ : 501.1373; found: 501.136.

Example 133: 2-[[6-(1,3-Benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]-(5-hydroxy-4methoxypentyl)amino]thiazol-4-carboxylic acid

Step A: Methyl 2-[tert-butoxycarbonyl-[5-[tert-butyl(dimethyl)silyl]oxy-4methoxypentyl]amino]thiazole-4-carboxylate.

Using the general Mitsunobu reaction procedure, starting from 571 mg of methyl 2-(tert-butoxycarbonylamino)thiazole-4-carboxylate (2.21 mmol, 1.1 eq.) and 500 mg of synthesis product 2b (2.01, 1 eq.) as the appropriate alcohol, 818 was obtained mg (83%) of the target product.

¹ H NMR (400 MHz, DMSO^) δ ppm 8.08 (s, 1H), 4.06 (t, 2H), 3.79 (s, 3H), 3.53 (ddd, 2H), 3.28 (s, 3H), 3.20-3.14 (m, 1H), 1.77-1.63 (m, 2h), 1.53 (s, 3H), 1.49-1.33 (m, 2H), 0.82 (s, 9H), -0.01 (s, 6H); ¹³ C NMR (100 MHz, DMSO^) δ ppm 161.4, 160.4, 140.6, 124.3, 80.4, 63.6, 56.9, 51.9, 46.8, 27.4, 27.6, 25.7, 23.4, 17.9 -5.5; LC-MS-ESI (m/z): [M+H] ⁺ , calculated for C ₂₂ H ₄₁ N ₂ O ₆ SSi: 489.2; found: 489.3.

Step B: methyl 2-[[5-[tert-butyl(dimethyl)silyl]oxy-4-methoxypentyl]amino]thiazole-4carboxylate.

Using the general HFIP deprotection procedure, starting from 306 mg of the product from Step A (0.63 mmol) as the appropriate carbamate, 234 mg (96%) of the desired product was obtained.

1H NMR (400 MHz, DMSO^) δ ppm 7.80 (t, 1H), 7.50 (s, 1H), 3.73 (s, 3H), 3.60-3.51 (m, 2H), 3.30 (s, 3H), 3.24-3.15 (m, 3H), 1.64-1.34 ( m, 4H), 0.85 (s, 9H), 0.03 (s, 6H); ¹³ C NMR (100 MHz, DMSO^) δ ppm 168.3, 161.6, 142.1, 116.4, 80.7, 64.3, 57.0, 51.6, 44.5, 25.8, 24.6, 17.9, -5.4; LC-MS-ESI (m/z): [M+H] ⁺ , calculated for C ₁₇ H ₃₃ N ₂ O ₄ SSi: 389.2; found: 389.3.

Step C: methyl 2-[[5-[tert-butyl(dimethyl)silyl]oxy-4-methoxypentyl]-[5-methyl-6-[^)-[3-(2

- 194 046336 trimethylsilylethoxymethyl)-1,3-benzothiazol-2-ylidene]amino]pyridazin-3-yl]amino]thiazol-4carboxylate.

Using the general Buchwald reaction procedure II, starting with 224 mg of the product from step B (0.58 mmol, 1 eq.) and 349 mg of the product of synthesis 4a (0.86 mmol, 1.5 eq.) as the appropriate halide, 357 mg (82%) were obtained target product.

¹ H NMR (500 MHz, DMSOC) δ ppm 7.98 (s, 1H), 7.83 (d, 1H), 7.69 (s, 1H), 7.46 (d, 1H), 7.43 (td, 1H), 7.25 (td, 1H), 5.86 (s, 2H), 4.46 (t, 2H), 3.81 (s, 3H), 3.72 (t, 2H), 3.56-3.51 (dd+dd, 2H), 3.29 (s, 3H), 3.22 (m, 1H), 2.45 (s, 3H ), 1.76 (m, 2H), 1.54-1.47 (m+m, 2H), 0.91 (t, 2H), 0.79 (s, 9H), -0.04 (s, 6H), -0.12 (s, 9H); ¹³ C NMR (125 MHz, DMSOC) δ ppm 162.3, 127.2, 123.4, 123.2, 123.1, 117.6, 112.0, 80.9, 72.9, 66.7, 64.2, 57.3, 52.2, 47.4, 28.1, 26.2, 23.3, 17.9, 17.8, -1. 0, -5.1; MSHR-ERI (m/z): [M+H] ⁺ , calculated for C35H55N6O5S ₂ Si2: 759.3208; found: 759.3212.

Step D: 2-[[6-(1,3-benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]-(5-hydroxy-4methoxypentyl)amino]thiazol-4-carboxylic acid.

Using the general deprotection and hydrolysis procedure followed by purification by reverse phase preparative chromatography (C18, 25 mM NH4HCO3 in water:MeCN), the desired product was obtained from the product from step C.

MSHR-ERI (m/z): [M+H] ⁺ , calculated for C ₂₂ H ₂₅ N ₆ O ₄ S ₂ : 501.1373; found: 501.1366.

Example 134: 2-[[6-(1,3-benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]-(4-hydroxybutyl)amino] 5-[3-[4-[3 -(dimethylamino )prop-1-ynyl]-2-fluorophenoxy]propyl]thiazole-4-carboxylic acid

Using the general deprotection and hydrolysis procedure starting from the appropriate methyl ester from Synthesis 5d, Step A, the desired product was obtained.

MSHR-ERI (m/z): [M+H] ⁺ , calculated for C ₃₄ H ₃₇ FN ₇ O ₄ S ₂ : 690.2327; found: 690.2347.

Example 135: 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)-5-{1[(tert-butoxy)carbonyl] azetidin-3-yl}-1,3-thiazol-4-carboxylic acid

Step A: ethyl 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)-5-{1[(tert-butoxy)carbonyl ]azetidin-3-yl}-1,3-thiazol-4-carboxylate.

To a solution of synthesis product 51, step A (122 mg, 0.17 mmol, 1 eq.) in tetrahydrofuran (6 ml) was added tetrabutylammonium fluoride (1 M in tetrahydrofuran; 0.24 ml, 1.38 mmol, 8 eq.), and the mixture was heated at 50°C for 13 hours. The reaction mixture was allowed to cool to ambient temperature, then partitioned between dichloromethane and saturated aqueous sodium bicarbonate, and the organic phase was washed with brine, dried (magnesium sulfate) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 24 g RediSep™ silica gel cartridge) eluting with a gradient of 0-10% methanol in dichloromethane gave the title product as a yellow glass (38 mg, 0.07 mmol, 38%) .

LC/MS (C ₂₇ H ₃₁ N ₇ O ₄ S ₂ ) 582 [M+H] ⁺ ; RT 1.26 (ZhKhMS-U-V1).

¹ H NMR (400 MHz, DMSO^) δ 11.05 (br s, 1H), 7.98 (br s, 1H), 7.81-7.61 (m, 2H), 7.50-7.33 (m, 1H), 7.31-7.16 (m , 1H), 4.65-4.53 (m, 1H), 4.37 (t, 2H), 4.29 (q, J=7.1 Hz, 2H), 3.98-3.84 (m, 2H), 3.78 (s, 3H), 2.47 ( s, 3H), 1.43 (s, 9H), 1.31 (t, 3H).

Step B: 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)-5-{ 1-[(tert-butoxy)carbonyl]azetidine -3-yl}-1,3-thiazol-4-carboxylic acid.

To a solution of the product from step A (38 mg, 0.07 mmol, 1 eq.) in 1,4-dioxane (3 ml) was added 2 M

- 195 046336 aqueous sodium hydroxide solution (0.5 ml, 1 mmol, 15 eq.), and the mixture was heated at 100°C for 30 minutes. The reaction mixture was allowed to cool to ambient temperature and concentrated in vacuo. Purification by reverse phase automated flash chromatography (CombiFlash Rf, C18 13 g RediSep column) eluting with a gradient of 5-95% acetonitrile in water gave the title product as a yellow solid (21.4 mg, 0.04 mmol, 59% ).

MSVR-ERI (m/z) [MH] ^- , calculated for C ₂₅ H ₂₆ N ₇ O ₄ S ₂ : 552.1493; found: 552.1511.

Example 136: 5-(1-Benzoylazetidin-3-yl)-2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3yl} (methyl)amino)-1, 3-thiazole-4-carboxylic acid

Step A: ethyl 5-(1-benzoylazetidin-3-yl)-2-[methyl(5-methyl-6-{[(2Z)-3-{[2(trimethylsilyl)ethoxy]methyl}-2,3- dihydro-1,3-benzothiazol-2-ylidene]amino}pyridazin-3-yl)amino]1,3-thiazol-4-carboxylate.

Triethylamine (0.01 ml, 0.1 mmol, 2 eq.) was added to a solution of synthesis product 51 (29.6 mg, 0.05 mmol, 1 eq.) and benzoyl chloride (6.74 μl, 0.06 mmol, 1.2 eq.) in dichloromethane (3 ml) and the mixture was stirred at ambient temperature for 3.5 h. Purification by automated flash column chromatography (CombiFlash Rf, 12 g RediSep™ silica gel cartridge) eluting with a gradient of 0-85% ethyl acetate in isoheptane gave the title product as a colorless glass substances (32 mg, 0.04 mmol, 92%).

LC/MS (C ₃₅ H ₄₁ N ₇ O ₄ SiS ₂ ) 716 [M+H]+; RT 1.59 (LCMS-V-BI).

¹ H NMR (400 MHz, DMSO) δ 7.86 (dd, J=7.6, 1.1 Hz, 1H), 7.75-7.67 (m, 3H), 7.58-7.39 (m, 5H), 7.297.22 (m, 1H) , 5.87 (s, 2H), 4.88-4.77 (m, 1H), 4.75-4.66 (m, 1H), 4.65-4.54 (m, 1H) 4.46-4.34 (m, 1H), 4.29 (q, J=7.1 Hz, 2H), 4.22-4.10 (m, 1H), 3.79 (s, 3H), 3.76-3.67 (m, 2H), 2.47 (d, J=1.0 Hz, 3H), 1.31 (t, J=7.1 Hz , 3H), 0.97-0.88 (m, 2H), -0.11 (s, 9H).

Step B: ethyl 5-(1-benzoylazetidin-3-yl)-2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin3-yl}(methyl)amino)-1 ,3-thiazole-4-carboxylate.

To a solution of the product from step A (32 mg, 0.04 mmol, 1 eq) in dichloromethane (3 ml) was added trifluoroacetic acid (0.45 ml, 6.05 mmol, 135 eq) and the mixture was stirred at ambient temperature for 5 h. The reaction mixture was partitioned between dichloromethane and saturated aqueous sodium bicarbonate, and the organic phase was washed with saturated aqueous sodium bicarbonate, then brine, dried (magnesium sulfate) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 12 g RediSep™ silica gel cartridge) eluting with a gradient of 0-5% methanol in dichloromethane gave the title product as a yellow glass (17.5 mg, 0.03 mmol, 67%) .

LC/MS (C ₂₉ H ₂₇ N ₇ O ₃ S ₂ ) 586 [M+H] ⁺ ; RT 1.30 (LCMS-V-BI).

1H NMR (400 MHz, DMSO-diO δ 7.94 (br s, 1H), 7.76-7.68 (m, 3H), 7.60-7.44 (m, 4H), 7.43-7.32 (m, 1H), 7.29-7.15 (m , 1H), 4.86-4.77 (m, 1H), 4.76-4.67 (m, 1H), 4.66-4.57 (m, 1H), 4.43-4.34 (m, 1H), 4.29 (q, 2H), 4.21-4.11 (m, 1H), 3.79 (s, 3H), 2.47 (s, 3H), 1.32 (t, J=7.1 Hz, 3H).

Step C: 5-(1-benzoylazetidin-3-yl)-2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3yl} (methyl)amino)-1, 3-thiazole-4-carboxylic acid.

To a solution of the product from step B (17.5 mg, 0.03 mmol, 1 eq.) in 1,4-dioxane (3 ml) was added 2 M aqueous sodium hydroxide (0.5 ml, 1 mmol, 33.5 eq.), and the mixture was heated with reflux for 2 hours. The reaction mixture was allowed to cool to ambient temperature and purified using reverse phase flash chromatography (CombiFlash Rf, C18 13 g RediSep column) eluting with a gradient of 5-95% acetonitrile in water to give the target product as a yellow solid (16 mg, 0.03 mmol, 96%) [as sodium salt].

MSVR-ERI (m/z) [MH] ^- , calculated for C ₂₇ H ₂₂ N ₇ O ₃ S ₂ : 556.1231; found: 556.1260.

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Example 137: 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)-5-(3-{4-[3(dimethylamino )prop-1-yn-1-yl]-2-fluorophenoxy}-2,2-dimethylpropyl)-1,3-thiazol-4-carboxylic acid

Step A: methyl 5-(3-{4-[3-(dimethylamino)prop-1-yn-1-yl]-2-fluorophenoxy}-2,2-dimethylpropyl)-2[methyl(5-methyl-6 -{[(2Z)-3-{[2-(trimethylsilyl)ethoxy]methyl}-2,3-dihydro-1,3-benzothiazol-2ylidene]amino}pyridazin-3-yl)amino]-1,3- thiazole-4-carboxylate.

To a solution of synthesis product 3zc (8 mg, 0.02 mmol, 1 eq.) in 1,4-dioxane (2 ml) was added synthesis product 4a (9.76 mg, 0.02 mmol, 1.3 eq.), xanthos (2.14 mg, 3.69 μmol, 0.2 equiv.), cesium carbonate (18 mg, 0.06 mmol, 3 equiv.) and CA'-diisopropylethylamine (9.64 μl, 0.06 mmol, 3 equiv.). The mixture was bubbled with nitrogen (10 min), then tris(dibenzylideneacetone)dipalladium(0) (1.69 mg, 0.1 eq.) was added and the mixture was heated at 120°C overnight. The reaction mixture was partitioned between ethyl acetate and brine, and the organic phase was dried (magnesium sulfate) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 4 g RediSep™ silica gel cartridge) eluting with a gradient of 0-6% methanol in dichloromethane provided the title product as a yellow solid (15 mg, 0.02 mmol, 100%) .

LC/MS (C ₄₀ H ₅₀ FN ₇ O ₄ SiS ₂ ) 804 [M+H]+; RT 2.65 (LCMS-VC).

¹ H NMR (400 MHz, DMSO^ ₆ ) δ 7.83 (d, J=7.9 Hz, 1H), 7.67 (d, J=1.2 Hz, 1H), 7.49-7.42 (m, 2H), 7.357.23 (m , 2H), 7.22-7.13 (m, 2H), 5.86 (s, 2H), 3.83 (s, 2H), 3.79-3.75 (m, 5H), 3.75-3.67 (m, 2H), 2.45 (s, 3H ), 2.15 (s, 6H), 1.05 (s, 6H), 0.96-0.88 (m, 2H), -0.11 (s, 9H).

Step B: 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)-5-(3-{4-[3(dimethylamino )prop-1-yn-1-yl]-2-fluorophenoxy}-2,2-dimethylpropyl)-1,3-thiazol-4-carboxylic acid.

To a solution of the product from step A (60 mg, 0.07 mmol, 1 eq.) in tetrahydrofuran (6 ml) was added ethylenediamine (15 μl, 0.22 mmol, 3 eq.) and tetrabutylammonium fluoride (1 M in tetrahydrofuran, 224 μl, 0.22 mmol , 3 eq.), and the mixture was heated at 70°C overnight. The solvent was removed in vacuo and the crude product was purified by reverse phase automatic flash chromatography (CombiFlash Rf, C18 13 g RediSep column) eluting with a gradient of 5-95% acetonitrile in water to give a solid, which was triturated with diethyl ether, filtered and dried in vacuo to give the title product as a yellow solid (13.1 mg, 0.02 mmol, 27%).

HRMS-ERI (m/z) [M+H]+, calculated for C ₃₃ H ₃₅ FN7O3S ₂ : 660.2221; found: 660.2264.

Example 138: 2-({6-[(1,3-benzothiazol-2-yl)amino]-5 -methylpyridazin-3 -yl} (methyl)amino)-5-[1-(2phenylacetyl)azetidine-3 - yl]-1,3-thiazole-4-carboxylic acid

Step A: ethyl 2-[methyl(5-methyl-6-{[(2Z)-3-{[2-(trimethylsilyl)ethoxy]methyl}-2,3-dihydro-1,3benzothiazol-2-ylidene]amino }pyridazin-3-yl)amino]-5-[1-(2-phenylacetyl)azetidin-3-yl]-1,3-thiazol-4carboxylate.

To a solution of synthesis product 51 (105 mg, 0.17 mmol, 1 eq.) in dichloromethane (6 ml) was added triethylamine (0.05 ml, 0.34 mmol, 2 eq.) and phenylacetyl chloride (0.03 ml, 0.21 mmol, 1.2 eq.), and the mixture was stirred at ambient temperature for 3 hours. The reaction mixture was diluted with dichloromethane, then washed with water and brine, dried (magnesium sulfate) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 12 g RediSep™ silica gel cartridge) eluting with a gradient of 0-100% ethyl acetate in isoheptane gave the title product as a yellow foam (66 mg, 0.09 mmol, 53%).

LC/MS (C ₃₆ H ₄₃ N ₇ O ₄ SiS ₂ ) 730 [M+H]+; RT 1.45 (LCMS-V-BI).

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1H NMR (400 MHz, DMSO-cf,) δ 7.85 (dd, J=7.8, 1.1 Hz, 1H), 7.70 (d, J=1.1 Hz, 1H), 7.52-7.39 (m, 2H), 7.34-7.25 (m, 5H), 7.24-7.18 (m, 1H), 5.87 (s, 2H), 4.74-4.58 (m, 2H), 4.39 (t, J=9.1 Hz, 1H), 4.29 (q, J=7.1 Hz, 2H), 4.23 (dd, J=8.1, 5.8 Hz, 1H), 3.96 (dd, J=9.7, 6.1 Hz, 1H), 3.79 (s, 3H), 3.77-3.68 (m, 2H), 3.51 (d, J=2.1 Hz, 2H), 2.47 (d, J=1.0 Hz, 3H), 1.32 (t, J=7.1 Hz, 3H), 0.98-0.87 (m, 2H), -0.11 (s, 9H ).

Step B: ethyl 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)-5-[1-(2phenylacetyl)azetidine-3 -yl]-1,3-thiazol-4-carboxylate.

To a solution of the product from Step A (66 mg, 0.09 mmol, 1 eq.) in dichloromethane (3 mL) was added trifluoroacetic acid (1 mL, 13.4 mmol, 148 eq.) and the mixture was stirred at ambient temperature overnight. The reaction mixture was partitioned between dichloromethane and saturated aqueous sodium bicarbonate, washed with saturated aqueous sodium bicarbonate, then brine, dried (magnesium sulfate) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 12 g RediSep™ silica gel cartridge) eluting with a gradient of 0-5% methanol in dichloromethane gave the title product as a yellow glass (39 mg, 0.07 mmol, 72%) .

LC/MS (C ₃₀ H ₂₉ N ₇ O ₃ S ₂ ) 600 [M+H]+; RT 1.33 (LCMS-V-Bi).

1H NMR (400 MHz, DMSO) δ 11.07 (br s, 1H), 8.00 (br s, 1H), 7.72 (br s + s, 2H), 7.40 (br s, 1H), 7.37-7.25 (m, 4H ), 7.27-7.18 (m, 1H), 4.75-4.57 (m, 2H), 4.40 (t, J=9.1 Hz, 1H), 4.30 (q, J=7.1 Hz, 2H), 4.24 (dd, J= 8.1, 5.9 Hz, 1H), 3.96 (dd, J=9.7, 6.1 Hz, 1H), 3.79 (s, 3H), 3.52 (s, 2H), 1.33 (t, J=7.1 Hz, 3H).

Step C: 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)-5-[1-(2phenylacetyl)azetidine-3- yl]-1,3-thiazole-4-carboxylic acid.

To a solution of the product from step B (39 mg, 0.07 mmol, 1 eq.) in 1,4-dioxane (4 ml) was added 2 M aqueous sodium hydroxide (1 ml, 2 mmol, 30.8 eq.), and the mixture was heated with reflux for 1.5 h. The solvent was removed in vacuo and the crude product was purified by reverse phase flash chromatography (CombiFlash Rf, C18 13 g RediSep column) eluting with a gradient of 5-95% acetonitrile in water to give the title product as yellow solid (28.7 mg, 0.05 mmol, 77%) [as sodium salt].

MSVR-ERI (m/z) [MH] ^- , calculated for C ₂₈ H ₂₄ N ₇ O ₃ S ₂ : 570.1388; found: 570.1423.

Example 139: 5-(Azetidin-3-yl)-2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3yl}(methyl)amino)-1,3- thiazole-4-carboxylic acid

In the preparation of Example 138, Step C, 5-(azetidin-3-yl)-2-({6-[(1,3-benzothiazol-2yl)amino]-5-methylpyridazin-3-yl}(methyl)amino Sodium )-1,3-thiazole-4-carboxylate (7.5 mg, 0.02 mmol, 25%) was isolated as a by-product [as sodium salt].

MSVR-ERI (m/z) [MH] ^- , calculated for C20H18N7O2S2: 452.0969; found: 452.0977.

Example 140: 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)-5-{[1-({4[3- (dimethylamino)prop-1-in-1-yl]-2-fluorophenoxy}methyl)cyclopropyl]methyl}-1,3-thiazol-4-carboxylic acid

\

Step A: methyl 5-{[1-({4-[3-(dimethylamino)prop-1-yn-1-yl]-2-fluorophenoxy}methyl)cyclopropyl]methyl}-2-[methyl(5-methyl -6-{[(27)-3-{[2-(trimethylsilyl)ethoxy]methyl}-2,3-dihydro-1,3benzothiazol-2-ylidene]amino}pyridazin-3-yl)amino]-1, 3-thiazole-4-carboxylate.

To a solution of synthesis product 3zd (84 mg, 0.19 mmol, 1 eq.) in 1,4-dioxane (6 ml) was added synthesis product 4a (103 mg, 0.25 mmol, 1.3 eq.), xanthos (22.5 mg, 0.04 mmol, 0.2 equiv.), cesium carbonate (190 mg,

- 198 046336

0.58 mmol, 3 equiv.) and N,N-diisopropylethylamine (102 μl, 0.58 mmol, 3 equiv.). The mixture was bubbled with nitrogen (10 min), then tris(dibenzylideneacetone)dipalladium(0) (17.8 mg, 0.02 mmol, 0.1 eq.) was added and the mixture was heated at 120°C overnight. The reaction mixture was partitioned between ethyl acetate and brine, and the organic phase was dried (magnesium sulfate) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 4 g RediSep™ silica gel cartridge) eluting with a gradient of 0-5% methanol in dichloromethane provided the title product as a brown solid (98 mg, 0.12 mmol, 63%) .

LC/MS (C ₄ 0H48FN7O ₄ SiS2) 802 [M+H]+; RT 2.63 (LCMS-VC).

1H NMR (400 MHz, DMSO-d^δ 7.87-7.80 (m, 1H), 7.67 (d, 1H), 7.52-7.40 (m, 2H), 7.33-7.22 (m, 2H), 7.18-7.11 (m , 1H), 7.03 (t, J=8.7 Hz, 1H), 5.87 (s, 2H), 3.83 (s, 2H), 3.76 (s, 3H), 3.76-3.69 (m, 2H), 3.66 (s, 3H), 3.39 (s, 2H), 3.36 (s, 2H), 2.46 (d, J=1.0 Hz, 3H), 2.18 (s, 6H), 0.97-0.88 (m, 2H), 0.78-0.65 (m , 4H), -0.10 (s, 9H).

Step B: methyl 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)-5-{[1({4-[3 -(dimethylamino)prop-1-in-1-yl]-2-fluorophenoxy}methyl)cyclopropyl]methyl}-1,3-thiazol-4carboxylate.

To a solution of the product from step A (98 mg, 0.12 mmol, 1 eq.) in tetrahydrofuran (6 ml) was added ethylenediamine (24.5 μl, 0.37 mmol, 3 eq.) and tetrabutylammonium fluoride (1 M in tetrahydrofuran; 224 μl, 0.37 ml , 0.37 mmol, 3 eq.), and the mixture was heated at 70°C overnight. The reaction mixture was partitioned between water and 9:1 dichloromethane/methanol, and the organic phase was separated and dried (PTFE phase separator), and concentrated in vacuo. Purification by reverse phase automated flash chromatography (CombiFlash Rf, C18 13 g RediSep column) eluting with a gradient of 5-95% acetonitrile in water gave the title product as a yellow solid (5.01 mg, 0.01 mmol, 6% ).

LC/MS (C ₃₄ H ₃₄ FN ₇ O ₃ S ₂ ) 672 [M+H] ⁺ ; RT 2.04 (LCMS-VC).

¹ H NMR (400 MHz, DMSO) δ 11.52 (br s, 1H), 7.95-7.88 (m, 1H), 7.69 (d, J=1.2 Hz, 1H), 7.54 (d, J=7.9 Hz, 1H) , 7.45-7.35 (m, 1H), 7.33-7.18 (m, 2H), 7.19-7.11 (m, 1H), 7.03 (t, J=8.8 Hz, 1H), 3.83 (s, 2H), 3.75 (s , 3H), 3.67 (s, 3H), 3.40 (s, 2H), 3.38 (s, 2H), 2.47 (d, J=1.1 Hz, 3H), 2.19 (s, 6H), 0.78-0.65 (m, 4H).

Step C: 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)-5-{[1-({4-[3 (dimethylamino)prop-1-in-1-yl]-2-fluorophenoxy}methyl)cyclopropyl]methyl}-1,3-thiazol-4-carboxylic acid.

To a solution of the product from step B (35 mg, 0.05 mmol, 1 eq.) in 1,4-dioxane (5 ml) was added lithium hydroxide monohydrate (21.9 mg, 0.52 mmol, 10 eq.), and the mixture was heated to 110°C during the night. The reaction mixture was concentrated in vacuo and the residue was triturated with water, filtered and dried in vacuo to give the title product as a yellow solid (23.8 mg, 0.04 mmol, 70%) [as lithium salt].

HRMS-ERI (m/z) [M+H] ⁺ , calculated for C ₃₃ H ₃₃ FN ₇ O ₃ S ₂ : 658.2065; found: 658.2109.

Example 141: 2-[[6-(1,3-benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]-(5-hydroxypentyl)amino]5-[3-[4-[3-(dimethylamino )prop-1-ynyl]-2-fluorophenoxy]propyl]thiazole-4-carboxylic acid

Using the general deprotection and hydrolysis procedure followed by repurification by reverse phase preparative chromatography (C18, 25 mM NH4HCO3 in water:MeCN), the desired product was obtained from synthesis 5f.

MSHR-ERI (m/z): [M+H] ⁺ , calculated for C ₃₅ H ₃₉ FN ₇ O ₄ S ₂ : 704.2489; found: 704.2483.

Example 142: 2-[[6-(1,3-Benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]-[5-[2(dimethylamino)ethylmethylamino]-4-hydroxypentyl]amino]thiazol-4 -carboxylic acid

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Step A: Methyl 2-[tert-butoxycarbonyl-[4-[tert-butyl(dimethyl)silyl]oxy-5-[2(dimethylamino)ethylmethylamino]pentyl]amino]thiazole-4-carboxylate.

Using the general Mitsunobu reaction procedure, starting with 426 mg of methyl 2-(tert-butoxycarbonylamino)thiazole-4-carboxylate (1.65 mmol, 1.1 eq.) and 478 mg of synthesis product 2f (1.65 mmol, 1 eq.) as the appropriate alcohol, we obtained 359 mg (43%) of target product.

1H NMR (400 MHz, DMSOC) δ ppm 8.08 (s, 1H), 3.79 (s, 3H), 3.72 (m, 1H), 2.44-2.20 (m, 8H), 2.18 (s, 6H), 2.14 (s, 3H), 1.85-1.56 (m, 3H), 1.53 (s, 9H), 1.39-1.25 (m, 1H), 0.79 (s, 9H), 0.01 (s, 3H), -0.02 (s, 3H); LC-MS-ESI (m/z): [M+H]+, calculated for C ₂₆ H ₅₁ N ₄ O ₅ SSi: 559.3; found: 559.4.

Step B: methyl 2-[[4-[tert-butyl(dimethyl)silyl]oxy-5-[2(dimethylamino)ethylmethylamino]pentyl]amino]thiazole-4-carboxylate.

Using the general HFIP deprotection procedure, starting from 811 mg of the product from Step A (1.45 mmol) as the appropriate carbamate, 262 mg (39%) of the desired product was obtained.

1H NMR (500 MHz, DMSOC) δ ppm 7.81 (t, 1H), 7.50 (s, 1H), 3.73 (s, 3H), 3.72 (m, 1H), 3.20 (m, 2H), 2.40 (m, 2H), 2.32 (m, 2H), 2.27 (d, 2H), 2.16 (s, 3H), 2.14 (s, 6H), 1.63-1.54 (m+m, 2H), 1.61-1.37 (m+m, 2H), 0.84 (s, 9H), 0.04 (s, 3H), 0.03 (s, 3H); ¹³ C NMR (125 MHz, DMSOC) δ ppm 168.7, 162.0, 142.6, 116.8, 70.4, 64.6, 57.4, 56.5, 52.0, 45.9, 45.1, 43.8, 33.0, 26.3, 24.9, -3.8, -4.3; LC-MS-ESI (m/z): [M+H] ⁺ , calculated for C21H43N4O ₃ SSi: 459.3; found: 459.4.

Step C: methyl 2-[[4-[tert-butyl(dimethyl)silyl]oxy-5-[2-(dimethylamino)ethylmethylamino]pentyl]-[5-methyl-6-[^)-[3-(2 -trimethylsilylethoxymethyl)-1,3-benzothiazol-2-ylidene]amino]pyridazin3-yl]amino]thiazol-4-carboxylate.

Using the general Buchwald reaction procedure II, starting with 255 mg of the product from step B (0.56 mmol, 1 eq.) and 226 mg of the product of synthesis 4a (0.77 mmol, 1.0 eq.) as the appropriate halide, 311 mg (67%) were obtained target product.

1H NMR (500 MHz, DMSOC) δ ppm 7.97 (s, 1H), 7.83 (dd, 1H), 7.70 (s, 1H), 7.46 (dd, 1H), 7.43 (m, 1H), 7.25 (m, 1H), 5.86 (s, 2H), 4.48 (t, 2H), 3.81 (s, 3H), 3.77 (m, 1H), 3.71 (t, 2H), 2.65 (m, 2H), 2.51 (m, 2H), 2.45 (s, 3H), 2.38 ( brs, 6h), 2.29 (m, 2H), 2.15 (s, 3H), 1.83-1.72 (m+m, 2H), 1.65-1.43 (m+m, 2H), 0.91 (t, 2H), 0.74 ( s, 9H), -0.01 & -0.06 (s+s, 6H), -0.12 (s, 9H); ¹³ C NMR (125 MHz, DMSOC) δ ppm 127.2, 123.4, 123.2, 123.1, 117.7, 112.0, 72.9, 69.9, 66.7, 64.0, 55.7, 54.9, 52.2, 47.3, 44.5, 43.2, 32.1, 26.1, 22.8, 17. 9, 17.8, 0.9, -3.9, -4.5; MSHR-ERI (m/z): [M+H] ⁺ , calculated for C ₃₉ H65N8O4S2Si2: 829.4103; found: 829.4096.

Step D: 2-[[6-(1,3-benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]-[5-[2(dimethylamino)ethylmethylamino]-4-hydroxypentyl]amino]thiazol-4 -carboxylic acid.

Using the general deprotection and hydrolysis procedure followed by purification by reverse phase preparative chromatography (C18, 25 mM NH4HCO3 in water:MeCN), the desired product was obtained from the product from step C.

MSHR-ERI (m/z): [M+H]+, calculated for C26H34N8O3S2: 571.2268; found: 571.2270.

Example 143: 2-[[6-(1,3-Benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]-[5-(dimethylamino)-4hydroxypentyl]amino]thiazol-4-carboxylic acid

Step A: Methyl 2-[tert-butoxycarbonyl-[4-[tert-butyl(dimethyl)silyl]oxy-5(dimethylamino)pentyl]amino]thiazole-4-carboxylate.

Using the general Mitsunobu reaction procedure, starting from 568 mg of methyl 2-(tert

- 200 046336 butoxycarbonylamino)thiazole-4-carboxylate (2.2 mmol, 1.1 eq.) and 523 mg of synthesis product 2d (2.0 mmol, 1 eq.) as a suitable alcohol, gave 530 mg (53%) of the target product.

¹ H NMR (500 MHz, DMSO-de) δ ppm 8.08 (s, 1H), 4.06 (m, 2H), 3.79 (s, 3H), 3.72 (m, 1H), 2.15 (d, 2H), 2.11 (s, 6H), 1.79-1.63 (m+m, 2H), 1.55-1.34 (m+m, 2H), 1.52 (s, 9H), 0.79 (s, 9H), 0.01 & -0.02 (s+s, 6H); ¹³ C NMR (125 MHz, DMSOC) δ ppm 160.8, 153.3, 124.7, 70.1, 65.9, 52.3, 47.3, 46.6, 32.7, 28.1, 26.2, 23.7, -3.8, 4.5; MSHR-ERI (m/z): [M+H] ⁺ , calculated for C ₂₃ H ₄₄ N ₃ O ₅ SSi: 502.2765; found: 502.2762.

Step B: methyl 2-[[4-[tert-butyl(dimethyl)silyl]oxy-5-(dimethylamino)pentyl]amino]thiazole-4carboxylate.

Using the general HFIP deprotection procedure, starting from 132 mg of the product from Step A (0.26 mmol) as the appropriate carbamate, 78 mg (74%) of the desired product was obtained.

1H NMR (500 MHz, DMSO-do) δ ppm 7.81 (t, 1H), 7.50 (s, 1H), 3.73 (s, 1H), 3.73 (s, 3H), 3.21 (q, 2H), 2.17 (d, 2H), 2.13 (s, 6H), 1.63 -1.55 (m+m, 2H), 1.60-1.37 (m+m, 2H), 0.84 (s, 9h), 0.04 & 0.03 (s+s, 6H); ¹³ C

NMR (125 MHz, DMSO-dg) δ ppm 168.7, 162.0, 116.9, 70.3, 66.3, 52.0, 46.6, 45.0, 33.1, 26.3, 25.0, -3.8, -4.3;

MSHR-ERI (m/z): [M+H] ⁺ , calculated for C18H36N3O3SSi: 402.2241; found: 402.2249.

Step C: methyl 2-[[4-[tert-butyl(dimethyl)silyl]oxy-5-(dimethylamino)pentyl]-[5-methyl-6-[^)[3-(2-trimethylsilylethoxymethyl)-1, 3-benzothiazol-2-ylidene]amino]pyridazin-3-yl]amino]thiazol-4carboxylate.

Using the general Buchwald reaction procedure II, starting with 72 mg of the product from step B (0.18 mmol, 1 eq.) and 73 mg of the product of synthesis 4a (0.18 mmol, 1.5 eq.) as the appropriate halide, 126 mg (91%) were obtained target product.

1H NMR (500 MHz, DMSOC) δ ppm 7.86-7.23 (4H), 7.97 (s, 1H), 7.69 (s, 1H), 5.87 (dd, 2H), 4.48 (m, 2H), 3.81 (s, 3H), 3.74 (m, 1H), 3.72 (t, 2H), 3.72 (t, 2H), 2.45 (s, 3H), 2.10 (brs, 6H), 1.83-1.71 (m+m, 2H), 1.59-1.41 (m+m, 2H), 0.92 (t, 2H), 0.74 (s, 9H), -0.01 & -0.06 (s+s, 6H), -0.12 (s, 9H); ¹³ C NMR (125 MHz,

DMSOC) δ ppm 123.1, 117.9, 72.9, 69.9, 66.8, 66.7, 52.2, 47.3, 46.6, 32.4, 26.2, 22.9, 17.8, 17.8, -1.0, -4.5;

MSHR-ERI (m/z): [M+H] ⁺ , calculated for C36H ₅₇ N ₇ O4S2Si2: 772.3525; found: 772.3521.

Step D: 2-[[6-(1,3-benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]-[5-(dimethylamino)-4hydroxypentyl]amino]thiazol-4-carboxylic acid.

Using the general deprotection and hydrolysis procedure followed by purification by reverse phase preparative chromatography (C18, 25 mM NH4HCO3 in water:MeC^, the desired product was obtained from the product from step C.

MSHR-ERI (m/z): [M+H] ⁺ , calculated for C23H28N7O3S2: 514.1690; found: 514.1694.

Example 144: 2-[[6-(1,3-Benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]-[4-hydroxy-5 (trimethylammonio)pentyl] amino]-5 -[3-[ 2-fluoro-4-[3-(methylamino)prop-1-ynyl]phenoxy]propyl]thiazole

4-carboxylate.

Step A: methyl 5-[3-[4-[3-[tert-butoxycarbonyl(methyl)amino]prop-1-ynyl]-2fluorophenoxy]propyl]-2-[[4-[tert-butyl(diphenyl)silyl ]oxy-5-(dimethylamino)pentyl]-[5-methyl-6-[^)[3-(2-trimethylsilylethoxymethyl)-1,3-benzothiazol-2-ylidene]amino]pyridazin-3-yl]amino] thiazole-4carboxylate.

Using the general tosylate alkylation procedure, starting from the synthesis product 5a and N methylmethanamine as a suitable amine, the desired product was obtained.

MSHR-ERI (m/z): [M+H] ⁺ , calculated for C64H ₈ 4FN ₈ O ₇ S2Si2: 1215.5421; found: 1215.5389.

Step B: 2-[[6-(1,3-benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]-[4-hydroxy-5(trimethylammonio)pentyl]amino]-5-[3-[ 2-fluoro-4-[3-(methylamino)prop-1-ynyl]phenoxy]propyl]thiazole 4-carboxylate.

The product from step A was dissolved in a mixture of acetonitrile (4 ml/mmol) and \,\-dimethylformamide (1 ml/mmol), then iodomethane (5 eq.) was added and the mixture was stirred at room temperature. until complete conversion was observed (approx. 1 hour). The reaction mixture was concentrated. A crude mixture which contained [5-[[5-[3-[4-[3-[tert-butoxycarbonyl(methyl)amino]prop-1-ynyl]-2-fluorophenoxy]propyl]-4methoxycarbonylthiazol-2-yl] -[5-methyl-6-[^)- [3 -(2-trimethylsilylethoxymethyl)-1, 3-benzothiazol-2

- 201 046336 ylidene]amino]pyridazin-3-yl]amino]-2-[tert-butyl(diphenyl)silyl]oxypentyl]trimethylammonium (LC-MS-ESI (m/z): [M] ⁺ , calculated for C ₆₅ H86FN ₈ O7S ₂ Si ₂ : 1229.6; found: 1229.4) was introduced into the next reaction step using the general procedure for deprotecting the quaternary salt to obtain the target product.

MSHR-ERI (m/z): [M+H] ⁺ , calculated for C ₃₇ H ₄₄ FN ₈ O ₄ S ₂ : 747.2905; found: 747.2900.

Example 145: 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)-5-(1benzylazetidin-3-yl)-1, 3-thiazole-4-carboxylic acid

Step A: ethyl 5-(1-benzylazetidin-3-yl)-2-[methyl(5-methyl-6-{[(2Z)-3-{[2(trimethylsilyl)ethoxy]methyl} -2,3- dihydro-1,3-benzothiazol-2-ylidene]amino}pyridazin-3-yl)amino]1,3-thiazol-4-carboxylate.

To a solution of synthesis product 51 (36.3 mg, 0.06 mmol, 1 eq.) in acetonitrile (3 ml) was added benzaldehyde (12.1 μl, 0.12 mmol, 2 eq.), sodium triacetoxyborohydride (37.7 mg, 0.18 mmol, 3 eq.) and glacial acetic acid (two drops), and the mixture was stirred at ambient temperature overnight. The reaction mixture was diluted with ethyl acetate and washed with saturated aqueous sodium bicarbonate, then brine, dried (magnesium sulfate) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 12 g RediSep™ silica gel cartridge) eluting with a gradient of 0-70% ethyl acetate in isoheptane gave the title product as a colorless gum (28.6 mg, 0.04 mmol, 69%).

LC/MS (C ₃₅ H ₄₃ N ₇ O ₃ SiS ₂ ) 702 [M+H] ⁺ ; RT 1.49 (LCMS-V-Bi).

¹ H NMR (400 MHz, DMSO^) δ 7.78 (dd, J=7.7, 1.1 Hz, 1H), 7.69 (d, J=1.1 Hz, 1H), 7.52-7.41 (m, 2H), 7.40-7.30 ( m, 4H), 7.29-7.20 (m, 2H), 5.88 (s, 2H), 4.46-4.35 (m, 1H), 4.27 (q, J=7.1 Hz, 2H), 3.78 (s, 3H), 3.773 .65 (m, 6H), 3.18 (dd, J=7.0, 5.9 Hz, 2H), 2.47 (d, J=1.0 Hz, 3H), 1.30 (t, J=7.1 Hz, 3H), 0.93 (dd, J=8.6, 7.4 Hz, 2H), -0.10 (s, 9H).

Step B: ethyl 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)-5-(1benzylazetidin-3-yl)-1 ,3-thiazole-4-carboxylate.

To a solution of the product from Step A (28.6 mg, 0.04 mmol, 1 eq.) in dichloromethane (3 mL) was added trifluoroacetic acid (0.3 mL, 4.03 mmol, 99 eq.) and the mixture was stirred at ambient temperature overnight. The reaction mixture was partitioned between dichloromethane and saturated aqueous sodium bicarbonate, and the organic phase was washed with saturated aqueous sodium bicarbonate, then brine, dried (magnesium sulfate) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 4 g RediSep™ silica gel cartridge) eluting with a gradient of 0-5% methanol in dichloromethane gave the title product as a yellow glass (17 mg, 0.03 mmol, 73%) .

LC/MS (C ₂₉ H ₂₉ N ₇ O ₂ S ₂ ) 572 [M+H] ⁺ ; RT 1.14 (LCMS-V-Bi).

1H NMR (400 MHz, DMSO^) δ 7.86 (br s, 1H), 7.71 (br s + s, 2H), 7.45-7.31 (m, 5H), 7.30-7.18 (m, 2H), 4.47-4.36 ( m, 1H), 4.27 (q, J=7.1 Hz, 2H), 3.78 (s, 3H), 3.73-3.61 (m, 4H), 3.19 (t, J=6.3 Hz, 2H), 2.48 (s, 3H ), 1.31 (t, J=7.1 Hz, 3H).

Step C: 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)-5-(1benzylazetidin-3-yl)-1, 3-thiazole-4-carboxylic acid.

To a solution of the product from step B (17 mg, 0.03 mmol, 1 eq.) in 1,4-dioxane (3 ml) was added 2 M aqueous sodium hydroxide (0.5 ml, 1 mmol, 33.6 eq.), and the mixture was heated with reflux for 2 hours. The reaction mixture was allowed to cool to ambient temperature and the solvent was removed in vacuo. Purification by reverse phase automated flash chromatography (CombiFlash Rf, C18 13 g RediSep column) eluting with a gradient of 5-95% acetonitrile in water gave the title product as a yellow solid (13.9 mg, 0.03 mmol, 86%) [ in the form of sodium salt].

HRMS-ERI (m/z) [M+H] ⁺ , calculated for C ₂₇ H ₂₆ N ₇ O ₂ S ₂ : 544.1584; found: 544.1616.

- 202 046336

Example 146: 3-[[5-[[6-(1,3-benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]-[4-carboxy-5-[3-[2fluoro-4-[ 3-(methylamino)prop-1-ynyl]phenoxy]propyl]thiazol-2-yl]amino]-2hydroxypentyl]dimethylammonio]propane-1-sulfonate.

The compound of Example 144, Step A, was suspended in MeCN (5 ml/mmol), then oxathiolane 2,2-dioxide (10 eq.) was added and the mixture was stirred at 60°C BH (complete conversion was observed). The reaction mixture was concentrated. Crude mixture which contained 3-[[5-[[5-[3-[4-[3-[tertbutoxycarbonyl(methyl)amino]prop-1-ynyl]-2-fluorophenoxy]propyl]-4-methoxycarbonylthiazol-2 -yl][5-methyl-6-[^)-[3-(2-trimethylsilylethoxymethyl)-1,3-benzothiazol-2-ylidene]amino]pyridazin-3yl]amino]-2-[tert-butyl(diphenyl )silyl]oxypentyl]dimethylammonio]propane-1-sulfonate (LC-MSERI (m/z): [M+H]+, calculated for C ₆₇ H ₉₀ FN ₈ O ₁₀ S ₃ Si ₂ : 1337.5; found: 1337.6) directly introduced into the next reaction step using the general quaternary salt deprotection procedure to obtain the desired product.

MSHR-ERI (m/z): [M+H] ⁺ , calculated for C ₃₉ H ₄₈ FN ₈ O ₇ S ₃ : 855.2787; found: 855.2786.

Example 147: 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(4hydroxybutyl)amino)-5-[3-(4-{ 3 -[ bis(3-sulfopropyl)amino]prop-1-yn-1-yl}-2-fluorophenoxy)propyl]1,3-thiazole-4-carboxylic acid

Example 147 was isolated as a by-product from the reaction of Example 148, Step B.

MSVR-ERI (m/z): [M+H] ⁺ , calculated for C ₃₈ H ₄₄ FN ₇ O ₁₀ S ₄ : 906.2089; found: 906.2098.

Example 148: 2-[[6-(1,3-benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]-(4-hydroxybutyl)amino] 5-[3-[2-fluoro-4-[ 3 -(3 -sulfopropylamino)prop-1-ynyl]phenoxy]propyl]thiazole-4-carboxylic acid

Step A: 5-[3-[4-(3-aminoprop-1-ynyl)-2-fluorophenoxy]propyl]-2-[[6-(1,3-benzothiazol-2ylamino)-5-methylpyridazin-3- yl]-(4-hydroxybutyl)amino]thiazole-4-carboxylic acid.

Using the general deprotection and hydrolysis procedure, starting from 370 mg of the product of synthesis 5i, step A (0.36 mmol, 1 eq.) 150 mg (62%) of the target product were obtained.

^1H NMR (500 MHz, DMSO-d^ δ ppm 7.88 (d, 1H), 7.66 (s, 1H), 7.53 (d, 1H), 7.38 (t, 1H), 7.22 (dd, 1H) , 7.22 (t, 1H), 7.16 (dd, 1H), 7.14 (t, 1H), 4.39 (t, 2H), 4.15 (t, 2H), 3.51 (s, 2H), 3.47 (t, 2H), 3.27 (t, 2H), 2.47 (s,

- 203 046336

3H), 2.14 (m, 2H), 1.74 (m, 2H), 1.52 (m, 2H); ¹³ C NMR (125 MHz, DMSO-de) δ ppm 126.7, 126.5, 122.6, 122.2, 119.1, 118.6, 116.8, 115.9, 91.0, 81.1, 68.8, 60.9, 47.2, 31.6, 31.0, 29.9, 24.2, 23.2, 17.7; MSVR-ERI (m/z): [M+H]+, calculated for C ₃₂ H ₃₃ FN ₇ O ₄ S ₂ : 662.2014; found: 662.2011.

Step B: 2-[[6-(1,3-benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]-(4-hydroxybutyl)amino]-5[3 - [2-fluoro-4-[ 3 -(3-sulfopropylamino)prop-1-ynyl]phenoxy]propyl]thiazole-4-carboxylic acid.

Oxathiolane 2,2-dioxide was added in one portion to a suspension of the product from step A in dry acetonitrile. The resulting mixture was stirred overnight at room temperature, and the disubstituted product (Example 147) was also formed. The crude product was dissolved in DMSO and then purified by reverse phase preparative chromatography (25 mM NH4HCO3 in water:MeCN) to give the title product.

MSVR-ERI (m/z): [M+H] ⁺ , calculated for C ₃₅ H ₃₉ FN ₇ O ₇ S ₃ : 784.2052; found: 784.2052.

Example 149: 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)-5-(1methanesulfonyl azetidin-3-yl)-1, 3-thiazole-4-carboxylic acid

Step A: ethyl 5-(1-methanesulfonyl azetidin-3-yl)-2-[methyl(5-methyl-6-{[(2Z)-3-{[2(trimethylsilyl)ethoxy]methyl}-2,3- dihydro-1,3-benzothiazol-2-ylidene]amino}pyridazin-3-yl)amino]1,3-thiazol-4-carboxylate.

To a solution of synthesis product 51 (24.6 mg, 0.04 mmol, 1 eq.) in dichloromethane (3 ml) was added methanesulfonyl chloride (3.73 μl, 0.05 mmol, 1.2 eq.) followed by the addition of a catalytic amount of 4-(dimethylamino)pyridine and triethylamine (0.01 ml, 0.08 mmol, 2 eq.), and the mixture was stirred at ambient temperature overnight.

The reaction mixture was partitioned between dichloromethane and saturated aqueous sodium bicarbonate, and the organic phase was washed with brine, dried (magnesium sulfate) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 4 g RediSep™ silica gel cartridge) eluting with a gradient of 0-100% ethyl acetate in isoheptane gave the title product as an off-white solid (16.4 mg, 0.02 mmol, 59 %).

LC/MS (C ₂₉ H ₃₉ N ₇ O ₅ SiS ₃ ) 690 [M+H] ⁺ ; RT 1.41 (LCMS-V-BI).

1H NMR (400 MHz, DMSO-dii) δ 7.87 (dd, J=7.6, 1.1 Hz, 1H), 7.71 (d, J=1.1 Hz, 1H), 7.51-7.39 (m, 2H), 7.29-7.22 (m , 1H), 5.87 (s, 2H), 4.61 (tt, J=8.4, 6.5 Hz, 1H), 4.39 (t, J=8.5 Hz, 2H), 4.30 (q, J=7.1 Hz, 2H), 3.93 (dd, J=8.3, 6.4 Hz, 2H), 3.80 (s, 3H), 3.76-3.67 (m, 2H), 3.10 (s, 3H), 2.47 (d, J=1.0 Hz, 3H), 1.32 ( t, J=7.1 Hz, 3H), 1.00-0.88 (m, 2H), -0.11 (s, 9H).

Step B: ethyl 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)-5-(1methanesulfonyl azetidin-3-yl)-1 ,3-thiazole-4-carboxylate.

To a solution of the product from Step A (16.4 mg, 0.02 mmol, 1 eq.) in dichloromethane (3 mL) was added trifluoroacetic acid (0.2 mL, 2.68 mmol, 113 eq.) and the mixture was stirred at ambient temperature overnight. The reaction mixture was diluted with dichloromethane, washed with saturated aqueous sodium bicarbonate (x2), then brine, dried (magnesium sulfate) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 4 g RediSep™ silica gel cartridge) eluting with a gradient of 0-10% methanol in dichloromethane provided the title product as a beige solid (10 mg, 0.02 mmol, 75%) .

LC/MS (C ₂₃ H ₂₅ N ₇ O ₄ S ₃ ) 560 [M+H] ⁺ ; RT 1.24 (LCMS-V-BI).

1H NMR (400 MHz, DMSO^) δ 7.92 (br s, 1H), 7.72 (s, 1H), 7.51-7.42 (m, 1H), 7.40-7.34 (m, 1H), 7.277.15 (m, 1H ), 4.68-4.54 (m, 1H), 4.49 (td, J=8.5, 2.9 Hz, 2H), 4.40 (t, 2H), 4.28 (q, 2H), 4.04 (dd, J=8.2, 6.5 Hz, 2H), 3.78 (s, 3H), 3.11 (s, 3H), 2.48 (s, 3H), 1.32 (t, J=7.1 Hz, 3H).

Step C: 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)-5-(1methanesulfonyl azetidin-3-yl)-1, 3-thiazole-4-carboxylic acid.

To a solution of the product from step B (10 mg, 0.02 mmol, 1 eq.) in 1,4-dioxane (2 ml) was added 2 M aqueous sodium hydroxide solution (0.2 ml, 0.4 mmol, 22.4 eq.), and the mixture was heated at 60 °C for 1.5 h. Purification by reverse phase automated flash chromatography (CombiFlash Rf, C18 13 g RediSep column) eluting with a gradient of 5-95% acetonitrile in water gave the title product as a yellow solid (7.6 mg, 0.01 mmol, 80%) [as sodium salt].

- 204 046336

MSHR-ERI (m/z) [M+H]+, calculated for C21H22N7O4S3: 532.0890; found: 532.0894.

Example 150: 2-[[6-(1,3-benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]-[4-hydroxy-5(methylamino)pentyl]amino]-5-[3-[ 4-[3-(dimethylamino)prop-1-ynyl]-2-fluorophenoxy]propyl]thiazole-4carboxylic acid

A general tosylate alkylation procedure was used starting from synthesis 5b and methanamine as a suitable amine, with the intermediate methyl 2-[[4-[tert-butyl(diphenyl)silyl]oxy-5-(methylamino)pentyl]-[5-methyl- 6-[^)-[3-(2-trimethylsilylethoxymethyl)1,3-benzothiazol-2-ylidene]amino]pyridazin-3-yl]amino]-5-[3-[4-[3-(dimethylamino)prop -1-ynyl]-2fluorophenoxy]propyl]thiazole-4-carboxylate was not isolated, and after evaporation of volatile substances, the general deprotection and hydrolysis procedure was used to obtain the target product.

MSHR-ERI (m/z): [M+2H] ²⁺ , calculated for C ₃₆ H ₄₃ FN ₈ O ₄ S ₂ : 367.1411; found: 367.1413.

Example 151: 2-[[6-(1,3-benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]-(4-hydroxybutyl)amino]5-[3-[2-fluoro-4-[ 3-[methyl (3-sulfopropyl)amino]prop-1-ynyl]phenoxy]propyl]thiazole-4-carboxylic acid

Step A: 3-[methyl(prop-2-ynyl)amino]propane-1-sulfonic acid.

To 0.34 ml of N-methylpropargylamine (4 mmol, 1 equiv.) in 1 ml of MeCN, 244 mg of oxathiolane 2,2 dioxide (2 mmol, 0.5 equiv.) was added, and the mixture was stirred at 60°C for 3 hours. After concentration, the residue was purified by flash column chromatography using DCM and MeOH as eluents to obtain 299 mg (78%) of the desired product.

¹ H NMR (400 MHz, DMSO-d,) δ ppm 10.10 (br, 1H), 4.01 (s, 2H), 3.75 (s, 1H), 3.12 (m, 2H), 2.72 (s, 3H), 2.59 (t, 2H), 1.94 (qui, 2H).

Step B: 3-[3-[4-[3-[2-[tert-butoxycarbonyl-[4-[tert-butyl(dimethyl)silyl]oxybutyl]amino]-4methoxycarbonylthiazol-5-yl]propoxy]-3- fluorophenyl]prop-2-ynylmethylamino]propane-1-sulfonic acid.

Using the general Sonogashira reaction procedure, starting with the product from step A and the product of synthesis 3c, step A, as starting materials, 303 mg (56%) of the target product was obtained.

1H NMR (400 MHz, DMSO-06) δ ppm 12.45 (vbrs, 1H), 7.45 (dd, 1H), 7.33 (dm, 1H), 7.18 (t, 1H), 4.24 (brs, 2H), 4.16-4.06 (m, 4H), 4.02 ( t, 2H), 3.75 (s, 3H), 3.58 (t, 2H), 3.22 (t, 2H), 2.80 (brs, 3H), 2.61 (brt, 2H), 2.08 (m, 2H), 1.97 (m , 2H), 1.68 (m, 2H), 1.57 (s, 9H), 1.43 (m, 2H), 0.82 (s, 9H), -0.01 (s, 6H).

Step C: 3-[3-[4-[3-[2-[4-[tert-Butyl(dimethyl)silyl]oxybutylamino]-4-methoxycarbonylthiazol5-yl]propoxy]-3-fluorophenyl]prop-2-ynylmethylamino ]propane-1-sulfonic acid.

Using the general HFIP deprotection procedure, starting with the product from Step B as the appropriate carbamate, 200 mg (81%) of the desired product was obtained.

1H NMR (500 MHz, DMSO-d,) δ ppm 10.38 (s, 1H), 7.59 (t, 1H), 7.47 (dd, 1H), 7.33 (m, 1H), 7.18 (t, 1H),

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4.28 (brs, 2H), 4.09 (t, 2H), 3.67 (s, 3H), 3.59 (t, 2H), 3.27 (brs, 2H), 3.17 (q, 2H), 3.12 (t, 2H), 2.82 (s, 3H), 2.62 (t, 2H), 2.00 (q, 2H), 2.00 (m, 2H), 1.54 (m, 2H), 1.49 (m, 2H), 0.84 (s, 9H), 0.02 ( s, 6H); ¹³ C NMR (125 MHz, DMSO-de) δ ppm 148.2, 129.6, 119.7, 115.4, 68.1, 62.6, 55.2, 51.7, 49.3, 45.8, 44.4, 40.0, 30.6, 30.2, 26.3, 25.6, 23.3, 21.0, -4.8.

Step D: 3-[3-[4-[3-[2-[4-[tert-butyl(dimethyl)silyl]oxybutyl-[5-methyl-6-[^)-[3-(2trimethylsilylethoxymethyl)-1 ,3-benzothiazol-2-ylidene]amino]pyridazin-3-yl]amino]-4methoxycarbonylthiazol-5-yl]propoxy]-3-fluorophenyl]prop-2-ynylmethylamino]propan-1-sulfonic acid.

Using the general Buchwald reaction procedure II, starting from the product from step C and the product of synthesis 4a as a suitable halide, 270 mg (88%) of the desired product were obtained.

1H NMR (500 MHz, DMSO^) δ ppm 7.84 (d, 1H), 7.66 (s, 1H), 7.47 (dm, 1H), 7.47 (dd, 1H), 7.44 (t, 1H), 7.33 (d, 1H), 7.26 (td, 1H), 7.21 (t, 1H), 5.86 (s, 2H), 4.42 (t, 2H), 4.24 (br., 2H), 4.17 (t, 2H), 3.77 (s, 3H), 3.72 (t, 2H), 3.61 (t, 2H), 3.29 (br., 2H), 3.27 (t, 2H), 2.79 (br., 3H), 2.60 (br., 2H), 2.45 (s, 3H), 2.13 (m, 2H) , 1.97 (br., 2H), 1.74 (m, 2H), 1.52 (m, 2H), 0.92 (t, 2H), 0.79 (s, 9H), -0.02 (s, 6H), -0.12 (s, 9H); ¹³ C NMR (125 MHz, DMSO^ ₆ ) δ ppm 129.5, 127.2, 123.5, 123.2, 119.7, 117.6, 115.4, 112.0, 72.9, 68.5, 66.7, 62.5, 51.9, 49.3, 46.5, 45.7, 40.0, 31.0, 29.7, 2 6.2, 23.8, 23.1, 21.1, 17.9, 17.8, -1.0, -4.9.

Step E: 2-[[6-(1,3-benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]-(4-hydroxybutyl)amino]-5[3-[2-fluoro-4-[ 3-[methyl(3-sulfopropyl)amino]prop-1-ynyl]phenoxy]propyl]thiazole-4-carboxylic acid.

The product from step D and LiOHxH ₂ O (10 eq.) in a 1:1 mixture of THF and water was kept at 50°C for 18 hours. After treating the reaction mixture with 2 ml of conc. HCl at 0°C, the reaction mixture was kept at 50°C for 1 hour. The reaction mixture was then poured onto cold ammonia solution, and the precipitated substance was filtered and purified using reverse phase preparative chromatography (25 mM NH4HCO3 in water:MeCN) to obtain the target product.

MSVR-ERI (m/z): [M+H]+, calculated for C3 ₆ H ₄₁ FN ₇ O7S3: 798.2214; found: 798.2204.

Example 152: 5-(1-Acetylazetidin-3-yl)-2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3yl}(methyl)amino)-1, 3-thiazole-4-carboxylic acid

Step A: ethyl 5-(1-acetylazetidin-3-yl)-2-[methyl(5-methyl-6-{[(2Z)-3-{[2(trimethylsilyl)ethoxy]methyl}-2,3- dihydro-1,3-benzothiazol-2-ylidene]amino}pyridazin-3-yl)amino]1,3-thiazol-4-carboxylate.

To a solution of synthesis product 51 (100 mg, 0.16 mmol, 1 eq.) in dichloromethane (5 ml) was added acetyl chloride (14 μl, 0.2 mmol, 1.2 eq.) and triethylamine (0.05 ml, 0.33 mmol, 2 eq.), and the mixture was stirred at ambient temperature for 4 hours. The reaction mixture was partitioned between dichloromethane and saturated aqueous sodium bicarbonate, and the organic phase was washed with brine, dried (magnesium sulfate) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 4 g RediSep™ silica gel cartridge) eluting with a gradient of 0-100% ethyl acetate in isoheptane gave the title product as a creamy foam (67.7 mg, 0.1 mmol, 63%).

LC/MS (C ₃₀ H ₃₉ N ₇ O ₄ SiS ₂ ) 654 [M+H]+; RT 1.37 (LCMS-V-BI).

1H NMR (400 MHz, DMSO^) δ 7.87 (dd, J=7.7, 1.1 Hz, 1H), 7.70 (d, J=1.2 Hz, 1H), 7.51-7.39 (m, 2H), 7.29-7.22 (m , 1H), 5.87 (s, 2H), 4.69-4.56 (m, 2H), 4.43-4.33 (m, 1H), 4.29 (q, 2H), 4.22-4.12 (m, 1H), 3.92 (dd, J =9.8, 5.3 Hz, 1H), 3.79 (s, 3H), 3.76-3.67 (m, 2H), 2.47 (d, J=1.0 Hz, 3H), 1.82 (s, 3H), 1.33 (t, J= 7.1 Hz, 3H), 0.97-0.88 (m, 2H), -0.11 (s, 9h).

Step B: ethyl 5-(1-acetylazetidin-3-yl)-2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3yl}(methyl)amino)-1 ,3-thiazole-4-carboxylate.

To a solution of the product from step A (67.7 mg, 0.1 mmol, 1 eq.) in tetrahydrofuran (3 ml) was added tetrabutylammonium fluoride (1 M in tetrahydrofuran; 0.04 ml, 0.31 mmol, 3 eq.) and ethylenediamine (0.02 ml, 0.31 mmol , 3 eq.), and the mixture was stirred at 60°C overnight. The reaction mixture was allowed to cool to ambient temperature, then partitioned between dichloromethane and saturated aqueous sodium bicarbonate, and the organic phase was washed with brine, dried (magnesium sulfate) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 12 g RediSep™ silica gel cartridge) eluting with a gradient of 0-5% methanol in dichloromethane gave the desired product as a yellow glassy solid.

- 206 046336 substance (10.5 mg, 0.02 mmol, 19%).

LC/MS (C ₂₄ H ₂₅ N ₇ O ₃ S ₂ ) 524 [M+H]+; RT 0.97 (LCMS-V-BI).

¹ HNMR (400 MHz, DMSO-de) δ 7.94 (br s, 1H), 7.72 (s, 1H), 7.55 (br s, 1H), 7.39 (t, J=7.5 Hz, 1H), 7.21 (t, J=7.6 Hz, 1H), 4.69-4.56 (m, 2H), 4.41-4.33 (m, 1H), 4.31 (q, 2H), 4.23-4.14 (m, 1H), 3.92 (dd, J=9.9, 5.3 Hz, 1H), 3.79 (s, 3H), 2.48 (s, 3H), 1.83 (s, 3H), 1.33 (t, J=7.1 Hz, 3H).

Step C: 5-(1-acetylazetidin-3-yl)-2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3 yl} (methyl)amino)-1 ,3-thiazole-4-carboxylic acid.

To a solution of the product from step B (10.5 mg, 0.02 mmol, 1 eq.) in 1,4-dioxane (3 ml) was added a 1 M aqueous solution of lithium hydroxide (0.1 ml, 0.1 mmol, 5 eq.), and the mixture was heated at 100°C overnight. Purification by reverse phase automated flash chromatography (CombiFlash Rf, C18 13 g RediSep column) eluting with a gradient of 5-95% acetonitrile in water gave the title product as a white solid (2 mg, 4.04 µmol, 20% ).

HRMS-ERI (m/z) [M+H]+, calculated for C ₂₂ H ₂₂ N ₇ O ₃ S ₂ : 496.1220; found: 496.1252.

Example 153: 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl} (methyl)amino)-5-(1 phenylazetidin-3-yl)-1 ,3-thiazole-4-carboxylic acid

Step A: ethyl 2-[methyl(5-methyl-6-{[(27)-3-{[2-(trimethylsilyl)ethoxy]methyl}-2,3-dihydro-1,3benzothiazol-2-ylidene] amino } pyridazin-3 -yl)amino]-5-(1-phenylazetidin-3-yl)-1,3-thiazol-4carboxylate.

Bromobenzene (20.6 μL, 0.2 mmol, 1.2 eq.), cesium carbonate (160 mg, 0.49 mmol, 3 eq.), N,Ndiisopropylethylamine (0.08 ml, 0.49 mmol, 3 eq.), xanthos (18.9 mg, 0.03 mmol, 0.2 eq.) and tris(dibenzylideneacetone) dipalladium(0) (15 mg, 0.02 mmol, 0.1 eq. ), and the mixture was refluxed overnight. The reaction mixture was partitioned between dichloromethane and water, and the organic phase was washed with brine, dried (magnesium sulfate) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 12 g RediSep™ silica gel cartridge) eluting with a gradient of 0-50% ethyl acetate in isoheptane provided the title product as a yellow solid (44.2 mg, 0.06 mmol, 39%) . LC/MS (C ₃₄ H ₄₁ N ₇ O ₃ SiS ₂ ) 688 [M+H] ⁺ ; RT 1.58 (LCMS-V-BI).

¹ H NMR (400 MHz, DMSO-dg) δ 7.82 (dd, J=7.7, 1.1 Hz, 1H), 7.69 (d, J=1.1 Hz, 1H), 7.49-7.35 (m, 2H), 7.28-7.17 (m, 3H), 6.74 (tt, J=7.4, 1.1 Hz, 1H), 6.64-6.49 (m, 2H), 5.85 (s, 2H), 4.81-4.69 (m, 1H), 4.38-4.23 (m , 4H), 3.86 (dd, J=7.3, 5.8 Hz, 2H), 3.79 (s, 3H), 3.76-3.67 (m, 2H), 2.45 (d, J=1.0 Hz, 3H), 1.34 (t, J=7.1 Hz, 3H), 0.96-0.87 (m, 2H), -0.12 (s, 9H).

Step B: ethyl 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)-5-(1phenylazetidin-3-yl)-1 ,3-thiazole-4-carboxylate.

To a solution of the product from step A (44 mg, 0.06 mmol, 1 eq.) in dichloromethane (5 ml) was added trifluoroacetic acid (0.5 ml, 6.71 mmol, 105 eq.) and the mixture was stirred at ambient temperature overnight. The reaction mixture was diluted with dichloromethane and washed with saturated aqueous sodium bicarbonate (x2), then brine, dried (magnesium sulfate) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 4 g RediSep™ silica gel cartridge) eluting with a gradient of 0-5% methanol in dichloromethane gave the title product as a yellow glass (18 mg, 0.03 mmol, 51%) .

LC/MS (C ₂₈ H ₂₇ N ₇ O ₂ S ₂ ) 558 [M+H] ⁺ ; RT 1.49 (LCMS-\'-B1).

¹ H NMR (400 MHz, DMSO-dg) δ 7.83 (br s, 1H), 7.70 (s, 1H), 7.52 (br s, 1H), 7.39-7.31 (m, 1H), 7.277.16 (m, 3H), 6.78-6.70 (m, 1H), 6.60-6.51 (m, 2H), 4.82-4.68 (m, 1H), 4.39-4.25 (m, 4H), 3.93-3.81 (m, 2H), 3.79 ( s, 3H), 2.47 (s, 3H), 1.34 (t, J=7.1 Hz, 3H).

Step C: 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)-5-(1phenylazetidin-3-yl)-1, 3-thiazole-4-carboxylic acid.

To a solution of the product from step B (18 mg, 0.03 mmol, 1 eq.) in 1,4-dioxane (3 ml) was added a 1 M aqueous solution of lithium hydroxide (0.16 ml, 0.16 mmol, 5 eq.), and the mixture was heated with reflux for 6 hours. Purification by reverse phase automated flash chromatography (CombiFlash Rf, C18 13 g RediSep column) eluting with a gradient of 5-95% acetonitrile in water gave the title product as a yellow solid (9.5 mg, 0.02 mmol, 56%)

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[as lithium salt].

HRMS-ERI (m/z) [M+H]+, calculated for C ₂₆ H ₂₄ N ₇ O ₂ S2: 530.1427; found: 530.1455.

Example 154: 5-[1-(Benzenesulfonyl)azetidin-3-yl]-2-({6-[(1,3-benzothiazol-2-yl)amino]-5methylpyridazin-3-yl}(methyl)amino) -1,3-thiazole-4-carboxylic acid

Step A: ethyl 5-[1-(benzenesulfonyl)azetidin-3-yl]-2-[methyl(5-methyl-6-{[(2Z)-3-{[2(trimethylsilyl)ethoxy]methyl}-2 ,3-dihydro-1,3-benzothiazol-2-ylidene]amino}pyridazin-3-yl)amino]1,3-thiazol-4-carboxylate.

To a solution of synthesis product 51 (46.7 mg, 0.08 mmol, 1 eq.) in dichloromethane (3 ml) was added benzenesulfonyl chloride (11.7 μl, 0.09 mmol, 1.2 eq.) followed by the addition of 4(dimethylamino)pyridine (2 mg, 0.02 mmol, 0.21 eq.) and triethylamine (0.02 ml, 0.15 mmol, 2 eq.), and the mixture was stirred at ambient temperature overnight. The reaction mixture was diluted with dichloromethane and washed with saturated aqueous sodium bicarbonate, then brine, then dried (magnesium sulfate) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 4 g RediSep™ silica gel cartridge) eluting with a gradient of 0-67% ethyl acetate in isoheptane provided the title product as a yellow glassy solid (40.3 mg, 0.05 mmol, 70%) .

LC/MS (C ₃₄ H ₄₁ N ₇ O ₅ SiS ₃ ) 752 [M+H] ⁺ ; RT 1.47 (LCMS-V-BI).

¹ H NMR (400 MHz, DMSO-06) δ 7.97-7.86 (m, 3H), 7.81-7.72 (m, 2H), 7.70-7.60 (m, 2H), 7.54-7.41 (m, 2H), 7.31- 7.25 (m, 1H), 5.89 (s, 2H), 4.52-4.39 (m, 1H), 4.29-4.18 (m, 4H), 3.79-3.67 (m, 7H), 2.47 (d, J=1.0 Hz, 3H), 1.27 (t, J=7.1 Hz, 3H), 0.98-0.89 (m, 2H), -0.10 (s, 9H).

Step B: ethyl 5-[1-(benzenesulfonyl)azetidin-3-yl]-2-({6-[(1,3-benzothiazol-2-yl)amino]-5methylpyridazin-3-yl}(methyl)amino )-1,3-thiazole-4-carboxylate.

To a solution of the product from Step A (40.3 mg, 0.05 mmol, 1 eq.) in dichloromethane (5 ml) was added trifluoroacetic acid (0.5 ml, 6.71 mmol, 125 eq.) and the mixture was stirred at ambient temperature overnight. The reaction mixture was diluted with dichloromethane and washed with saturated aqueous sodium bicarbonate (x2), then brine, dried (magnesium sulfate) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 4 g RediSep™ silica gel cartridge) eluting with a gradient of 0-6% methanol in dichloromethane provided the title product as a yellow solid (20 mg, 0.03 mmol, 60%) .

LC/MS (C ₂₈ H ₂₇ N ₇ O ₄ S ₃ ) 622 [M+H] ⁺ ; RT 1.37 (LCMS-V-BI).

1H NMR (400 MHz, DMSO-06) δ 8.00 (br s, 1H), 7.94-7.88 (m, 2H), 7.77 (t, J=7.6 Hz, 2H), 7.72-7.64 (m, 2H), 7.56 (br s, 1H), 7.40 (t, 1H), 7.24 (t, J=7.6 Hz, 1H), 4.46 (p, J=7.6 Hz, 1H), 4.30-4.18 (m, 4H), 3.81-3.66 (m, 5H), 2.45 (s, 3H), 1.26 (t, 3H).

Step C: 5-[1 -(benzenesulfonyl)azetidin-3-yl]-2-({6-[(1,3-benzothiazol-2-yl)amino]-5 methylpyridazin-3-yl}(methyl)amino )-1,3-thiazole-4-carboxylic acid.

To a solution of the product from step B (20 mg, 0.03 mmol, 1 eq.) in 1,4-dioxane (3 ml) was added a 1 M aqueous solution of lithium hydroxide (0.16 ml, 0.16 mmol, 5 eq.), and the mixture was heated at 100°C overnight. Purification by reverse phase automated flash chromatography (CombiFlash Rf, C18 15.5 g Gold RediSep column) eluting with a gradient of 5-95% acetonitrile in water gave the title product as a cream solid (4.9 mg, 0.01 mmol, 26 %).

HRMS-ERI (m/z) [M+H] ⁺ , calculated for C ₂₆ H ₂₄ N ₇ O ₄ S ₃ : 594.1046; found: 594.1060.

Example 155: 5-(1-Benzenesulfonamido-3-hydroxypropan-2-yl)-2-({6-[( 1,3-benzothiazol-2yl)amino]-5-methylpyridazin-3-yl}(methyl)amino )-1,3-thiazole-4-carboxylic acid

5-(1-Benzenesulfonamido-3-hydroxypropan-2-yl)-2-({6-[(1,3-benzothiazol-2-yl)amino]-5-208 046336 methylpyridazin-3-yl}(methyl) amino)-1,3-thiazole-4-carboxylic acid (2.9 mg, 4.74 µmol, 15%) was isolated as a by-product during the purification of Example 154, Step C.

HRMS-ERI (m/z) [M+H]+, calculated for C ₂₆ H ₂₆ N ₇ O ₅ S ₃ : 612.1152; found: 612.1154

Example 156: 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)-5-[1-(2methylpropyl)azetidine-3 - yl]-1,3-thiazol-4-carboxylic acid

Step A: ethyl 2-[methyl(5-methyl-6-{[(2Z)-3-{[2-(trimethylsilyl)ethoxy]methyl}-2,3-dihydro-1,3benzothiazol-2-ylidene] amino } pyridazin-3 -yl)amino]-5-[1 -(2-methylpropyl)azetidin-3-yl]-1,3-thiazol-4carboxylate.

To a solution of synthesis product 51 (134 mg, 0.22 mmol, 1 eq.) in a 2:1 mixture of acetonitrile/dichloromethane (6 ml) was added isobutyraldehyde (0.04 ml, 0.44 mmol, 2 eq.) followed by the addition of sodium triacetoxyborohydride (139 mg, 0.66 mmol, 3 eq.) and glacial acetic acid (10 μl), and the mixture was stirred at ambient temperature for 4.5 h. The reaction mixture was partitioned between dichloromethane and saturated aqueous sodium bicarbonate, and the organic phase was washed with brine, dried (sulfate magnesium) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 12 g RediSep™ silica gel cartridge) eluting with a gradient of 0-60% ethyl acetate in isoheptane gave the title product as a creamy foam (81.5 mg, 0.12 mmol, 56%).

LC/MS (C ₃₂ H ₄₅ N ₇ O ₃ SiS ₂ ) 668 [M+H] ⁺ ; RT 1.25 (LCMS-V-BI).

¹ H NMR (400 MHz, DMSO-do) δ 7.85-7.76 (m, 1H), 7.68 (d, 1H), 7.51-7.39 (m, 2H), 7.29-7.23 (m, 1H), 5.87 (s, 2H), 4.44-4.32 (m, 1H), 4.27 (q, J=7.1 Hz, 2H), 3.77 (s, 3H), 3.75-3.70 (m, 2H), 3.64 (t, J=7.0 Hz, 2H ), 3.09 (t, J=6.4 Hz, 2H), 2.46 (d, J=1.0 Hz, 3H), 2.26 (d, J=7.1 Hz, 2H), 1.58 (hept, J=6.8 Hz, 1H), 1.31 (t, J=7.1 Hz, 3H), 0.98-0.86 (m, 8H), -0.11 (s, 9H).

Step B: ethyl 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)-5-[1-(2methylpropyl)azetidine-3 -yl]-1,3-thiazol-4-carboxylate.

To a cooled solution of the product from Step A (81.5 mg, 0.12 mmol, 1 eq.) in dichloromethane (5 ml) was added trifluoroacetic acid (0.5 ml, 6.71 mmol, 55 eq.) and the mixture was stirred at ambient temperature overnight. The reaction mixture was quenched by adding saturated aqueous sodium bicarbonate, then extracted with dichloromethane and the organic extract was washed with brine, dried (magnesium sulfate) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 12 g RediSep™ silica gel cartridge) eluting with a gradient of 0-5% methanol in dichloromethane provided the title product as a yellow solid (50 mg, 0.09 mmol, 76%) .

LC/MS (C ₂₆ H ₃₁ N ₇ O ₂ S ₂ ) 538 [M+H] ⁺ ; RT 0.91 (LCMS-V-BI).

1H NMR (400 MHz, DMSO^) δ 7.87 (s, 1H), 7.70 (s, 1H), 7.52 (br s, 1H), 7.40 (t, J=7.6 Hz, 1H), 7.22 (t, J= 7.5 Hz, 1H), 4.44-4.33 (m, 1H), 4.28 (q, J=7.1 Hz, 2H), 3.77 (s, 3H), 3.64 (t, J=7.0 Hz, 2H), 3.10 (t, J=6.3 Hz, 2H), 2.47 (s, 3H), 2.27 (d, J=7.0 Hz, 2H), 1.66-1.51 (m, 1H), 1.32 (t, J=7.1 Hz, 3H), 0.93 ( d, J=6.6 Hz, 6H).

Step C: 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)-5-[1-(2methylpropyl)azetidine-3 - yl]-1,3-thiazol-4-carboxylic acid.

To a solution of the product from step B (50 mg, 0.09 mmol, 1 eq.) in 1,4-dioxane (3 ml) was added a 1 M aqueous solution of lithium hydroxide (0.46 ml, 0.46 mmol, 5 eq.), and the mixture was heated at 100°C overnight. Purification by reverse phase automated flash chromatography (CombiFlash Rf, C18 13 g RediSep column) eluting with a gradient of 5-95% acetonitrile in water gave the title product as a yellow solid (34.2 mg, 0.07 mmol, 72% ).

HRMS-ERI (m/z) [M+H] ⁺ , calculated for C ₂₄ H ₂₈ N ₇ O ₂ S ₂ : 510.1740; found: 510.1776.

Example 157: 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)-5-(1methylazetidin-3-yl)-1, 3-thiazole-4-carboxylic acid

- 209 046336

Step A: ethyl 2-[methyl(5-methyl-6-{[(2Z)-3-{[2-(trimethylsilyl)ethoxy]methyl}-2,3-dihydro-1,3benzothiazol-2-ylidene] amino } pyridazin-3-yl)amino]-5-(1-methylazetidin-3-yl)-1,3-thiazol-4carboxylate.

To a solution of synthesis product 51 (134 mg, 0.22 mmol, 1 eq.) in a 2:1 mixture of acetonitrile/dichloromethane (6 ml) was added formaldehyde (37% in water; 0.03 ml, 0.44 mmol, 2 eq.) followed by the addition of triacetoxyborohydride sodium (139 mg, 0.66 mmol, 3 eq.) and glacial acetic acid (10 μl), and the mixture was stirred at ambient temperature for 4.5 h. The reaction mixture was diluted with dichloromethane, then washed with saturated aqueous sodium bicarbonate followed by a wash with brine , dried (magnesium sulfate) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 12 g RediSep™ silica gel cartridge) eluting with a gradient of 0-100% ethyl acetate in isoheptane provided the title product as a cream solid (65 mg, 0.1 mmol, 47%) .

LC/MS (C ₂₉ H ₃₉ N ₇ O ₃ SiS ₂ ) 626 [M+H]+; RT 1.18 (LCMS-V-BI).

1H NMR (400 MHz, DMSOM ₆ ) δ 7.89-7.82 (m, 1H), 7.68 (d, J=1.2 Hz, 1H), 7.53-7.39 (m, 2H), 7.28-7.22 (m, 1H), 5.86 (s, 2H), 4.42-4.30 (m, 1H), 4.27 (q, J=7.1 Hz, 2H), 3.77 (s, 3H), 3.76-3.69 (m, 2H), 3.65 (t, J=7.2 Hz, 2H), 3.18-3.10 (m, 2H), 2.46 (d, J=1.0 Hz, 3H), 2.29 (s, 3H), 1.31 (t, J=7.1 Hz, 3H), 0.97-0.86 (m , 2H), -0.11 (s, 9H).

Step B: ethyl 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)-5-(1methylazetidin-3-yl)-1 ,3-thiazole-4-carboxylate.

To a cooled solution of the product from Step A (65 mg, 0.1 mmol, 1 eq.) in dichloromethane (3 mL) was added trifluoroacetic acid (0.3 mL, 4.03 mmol, 38.8 eq.) and the mixture was stirred at ambient temperature overnight. The reaction mixture was quenched by adding saturated aqueous sodium bicarbonate, then extracted with dichloromethane and the organic extract was washed with brine, dried (magnesium sulfate) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 12 g RediSep™ silica gel cartridge) eluting with a gradient of 0-10% methanol in dichloromethane gave the title product as a yellow solid (40 mg, 0.08 mmol, 78%) .

LC/MS (C ₂₃ H ₂₅ N ₇ O ₂ S ₂ ) 496 [M+H]+; RT 0.80 (LCMS-V-BI).

1H NMR (400 MHz, DMSO) δ 7.92 (d, J=7.8 Hz, 1H), 7.69 (d, J=1.2 Hz, 1H), 7.53 (s, 1H), 7.44-7.32 (m, 1H), 7.25 -7.16 (m, 1H), 4.42-4.30 (m, 1H), 4.27 (q, J=7.1 Hz, 2H), 3.76 (s, 3H), 3.65 (t, J=7.2 Hz, 2H), 3.15 ( t, 2H), 2.47 (d, J=1.0 Hz, 3H), 2.29 (s, 3H), 1.31 (t, J=7.1 Hz, 3H).

Step C: 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl} (methyl)amino)-5 -(1methylazetidin-3-yl)-1, 3-thiazole-4-carboxylic acid.

To a solution of the product from step B (40 mg, 0.08 mmol, 1 eq.) in 1,4-dioxane (3 ml) was added a 1 M aqueous solution of lithium hydroxide (0.4 ml, 0.4 mmol, 5 eq.), and the mixture was heated at 100°C overnight. Purification by reverse phase automated flash chromatography (CombiFlash Rf, C18 13 g RediSep column) eluting with a gradient of 5-95% acetonitrile in water gave the title product as a yellow solid (15.5 mg, 0.03 mmol, 41% ).

HRMS-ERI (m/z) [M+H]+, calculated for C ₂₁ H ₂₂ N ₇ O ₂ S ₂ : 468.1271; found: 468.1301.

Example 158: 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(3carboxypropyl)amino)-1,3-thiazol-4-carboxylic acid

Step A: ethyl 2-{[4-(tert-butoxy)-4-oxobutyl](5-methyl-6-{[(2Z)-3-{[2(trimethylsilyl)ethoxy]methyl}-2,3- dihydro-1,3-benzothiazol-2-ylidene]amino}pyridazin-3-yl)amino}1,3-thiazol-4-carboxylate.

To a solution of synthesis product 3ze (1.74 g, 5.53 mmol, 1.5 equiv.) in 1,4-dioxane (60 ml) was added synthesis product 4a (1.5 g, 3.69 mmol, 1 equiv.), N,N-diisopropylethylamine (1.93 ml , 11.1 mmol, 3 eq.), cesium carbonate (3.6 g, 11.1 mmol, 3 eq.) and xantphos (427 mg, 0.74 mmol, 0.2 eq.), and the mixture was bubbled with nitrogen (10 min). Tris(dibenzylideneacetone)dipalladium(0) (338 mg, 0.37 mmol, 0.1 eq.) was added and the mixture was heated in a stoppered flask at 140°C for 2 h. The reaction mixture was allowed to cool to ambient temperature, then partitioned between water and ethyl acetate, aqueous extract phase

- 210 046336 was diluted with ethyl acetate (3x80 ml) and the combined organic extracts were washed with brine, dried (magnesium sulfate) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 40 g RediSep™ silica gel cartridge) eluting with a gradient of 0-25% ethyl acetate in isoheptane provided the title product as a pale yellow solid (0.93 g, 1.36 mmol, 37%) .

LC/MS (C ₃₂ H ₄₄ N ₆ O ₅ SiS ₂ ) 685 [M+H]+; RT 1.75 (LCMS-V-BI).

¹ H NMR (400 MHz, DMSO-d^ δ 7.96 (s, 1H), 7.87-7.92 (m, 1H), 7.76 (d, J=1.2 Hz, 1H), 7.47-7.40 (m, 2H), 7.28 -7.22 (m, 1H), 5.878 (s, 2H), 4.42 (t, J=7.4 Hz, 2H), 4.28 (q, J=7.1 Hz, 2H), 3.79-3.67 (m, 2H), 2.47 ( d, J=1.0 Hz, 3H), 2.36 (t, J=7.3 Hz, 2H), 1.95 (p, J=7.4 Hz, 2H), 1.37 (s, 9H), 1.31 (t, J=7.1 Hz, 3H), 0.96-0.88 (m, 2H), -0.11 (s, 9H).

Step B: 4-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}[4-(ethoxycarbonyl)-1,3thiazol-2-yl]amino)butane acid.

To a solution of the product from step A (929 mg, 1.36 mmol, 1 eq) in dichloromethane (45 ml) was added trifluoroacetic acid (8.31 ml, 109 mmol, 80 eq) and the mixture was stirred at ambient temperature for 4 h. The reaction mixture was quenched by adding saturated aqueous sodium bicarbonate, then extracted with dichloromethane (3x40 ml) and the combined organic extracts were washed with brine, dried (magnesium sulfate) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 24 g RediSep™ silica gel cartridge) eluting with a gradient of 0-20% methanol in dichloromethane gave the title product as a yellow solid (186.8 mg, 0.37 mmol, 28%) .

LC/MS (C ₂₂ H ₂₂ N ₆ O ₄ S ₂ ) 499 [M+H] ⁺ ; RT 1.27 (LCMS^-B1).

^1H NMR (400 MHz, DMSO^ ₆ ) δ 9.23 (br s, 2H), 7.97 (s, 1H), 7.95-7.87 (m, 1H), 7.82 (s, 1H), 7.54 (br s, 1H) , 7.42-7.36 (m, 1H), 7.25-7.18 (m, 1H), 4.39 (t, J=7.7 Hz, 2H), 4.29 (q, J=7.1 Hz, 2H), 2.47 (s, 3H), 2.39 (t, J=7.2 Hz, 2H), 2.01-1.98 (m, 2H), 1.31 (t, J=7.1 Hz, 2H).

Step C: 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(3-carboxypropyl)amino)1,3-thiazol-4-carboxylic acid.

To a solution of the product from step B (80 mg, 0.16 mmol, 1 eq.) in 1,4-dioxane (10 ml) was added lithium hydroxide monohydrate (13.5 mg, 0.32 mmol, 2 eq.) and the mixture was refluxed at for 2 hours. The reaction mixture was concentrated in vacuo and purified using reverse phase automatic flash chromatography (CombiFlash Rf, C18 13 g RediSep column) eluting with 5-95% acetonitrile in water to obtain the crude product. It was dissolved in methanol with a few drops of triethylamine and then loaded onto a methanol-conditioned PE-AX cartridge, washed successively with methanol and dichloromethane, then eluted with 10% formic acid in dichloromethane and concentrated in vacuo to give the title product as a yellow solid (28.9 mg, 0.06 mmol, 38%).

HRMS-ERI (m/z) [M+H] ⁺ , calculated for C ₂₀ H ₁₉ N ₆ O ₄ S ₂ : 471.0904; found: 471.0915.

Example 159: 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3 -yl} (methyl)amino)-5-[1 (benzylcarbamoyl)azetidin-3 -yl ]-1,3-thiazole-4-carboxylic acid

Step A: ethyl 5-[1-(benzylcarbamoyl)azetidin-3-yl]-2-[methyl(5-methyl-6-{[(2Z)-3-{[2(trimethylsilyl)ethoxy]methyl}-2 ,3-dihydro-1,3-benzothiazol-2-ylidene]amino}pyridazin-3-yl)amino]1,3-thiazol-4-carboxylate.

To a solution of synthesis product 51 (150 mg, 0.25 mmol, 1 eq.) in dichloromethane (5 ml) was added benzyl isocyanate (0.04 ml, 0.29 mmol, 1.2 eq.) followed by the addition of triethylamine (0.07 ml, 0.49 mmol, 2 eq. ), and the mixture was stirred at ambient temperature overnight. The reaction mixture was partitioned between dichloromethane and saturated aqueous ammonium chloride, and the organic phase was washed with brine, dried (magnesium sulfate) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 12 g RediSep™ silica gel cartridge) eluting with a gradient of 0-100% ethyl acetate in isoheptane provided the title product as a yellow solid (125 mg, 0.17 mmol, 68%) .

LC/MS (C ₃ 6H44N8O4SiS ₂ ) 745 [M+H] ⁺ ; RT 1.43 (LCMS^-B1).

¹ H NMR (400 MHz, DMSO-d ₆ ) δ 7.84 (dd, J=7.5, 1.1 Hz, 1H), 7.70 (d, J=1.2 Hz, 1H), 7.51-7.39 (m, 2H), 7.35- 7.20 (m, 5H), 7.19 (td, J=6.3, 2.8 Hz, 1H), 7.08 (t, J=6.0 Hz, 1H), 5.87 (s, 2H), 4.64-4.51 (m, 1H), 4.36 (t, J=8.4 Hz, 2H), 4.29 (q, J=7.1 Hz, 2H), 4.24 (d, J=6.0 Hz, 2H), 3.88 (dd, J=8.2, 6.0 Hz, 2H), 3.79 (s, 3H), 3.76-

- 211 046336

3.69 (m, 2H), 2.47 (d, J=1.0 Hz, 3H), 1.32 (t, J=7.1 Hz, 3H), 0.97-0.88 (m, 2H), -0.11 (s, 9H).

Step B: ethyl 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)-5-[1(benzylcarbamoyl)azetidine-3 - yl]-1,3-thiazol-4-carboxylate.

To a solution of the product from Step A (125 mg, 0.17 mmol, 1 eq.) in dichloromethane (5 mL) was added trifluoroacetic acid (0.7 mL, 9.39 mmol, 56 eq.) and the mixture was stirred at ambient temperature overnight. The reaction mixture was quenched by adding saturated aqueous sodium bicarbonate, then extracted with dichloromethane, and the organic extract was washed with brine, dried (magnesium sulfate) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 12 g RediSep™ silica gel cartridge) eluting with a gradient of 0-9% methanol in dichloromethane gave the title product as a yellow glass (87.6 mg, 0.14 mmol, 85%) .

LC/MS (C ₃₀ H ₃₀ N8O ₃ S2) 615 [M+H]+; RT 1.10 (LCMS-V-BI).

¹ H NMR (400 MHz, DMSO^) δ 11.04 (br s, 1H), 7.95 (br s, 1H), 7.73-7.62 (m, 1H), 7.50-7.34 (m, 1H), 7.34-7.15 (m , 7H), 7.07 (t, J=6.0 Hz, 1H), 4.62-4.52 (m, 1H), 4.36 (t, 2H), 4.29 (q, 2H), 4.24 (d, 2H), 3.94-3. 83 (m, 2H), 3.77 (s, 3H), 2.47 (d, 3H), 1.32 (t, J=7.1 Hz, 3H).

Step C: 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3 -yl} (methyl)amino)-5-[ 1 (benzylcarbamoyl)azetidin-3 -yl ]-1,3-thiazole-4-carboxylic acid.

To a solution of the product from step B (87.6 mg, 0.14 mmol, 1 eq.) in 1,4-dioxane (5 ml) was added a 1 M aqueous solution of lithium hydroxide (0.71 ml, 0.71 mmol, 5 eq.), and the mixture was heated at 100°C overnight. Purification by reverse phase automated flash chromatography (CombiFlash Rf, C18 13 g RediSep column) eluting with a gradient of 5-95% acetonitrile in water gave the title product as a yellow solid (25.6 mg, 0.04 mmol, 31% ).

HRMS-ERI (m/z) [M+H] ⁺ , calculated for C ₂₈ H ₂₇ N ₈ O ₃ S ₂ : 587.1642; found: 587.1669.

Example 160: 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)-5-[1(ethylcarbamoyl)azetidin-3-yl ]-1,3-thiazole-4-carboxylic acid

Step A: ethyl 5-[1-(ethylcarbamoyl)azetidin-3-yl]-2-[methyl(5-methyl-6-{[(2Z)-3-{[2(trimethylsilyl)ethoxy]methyl}-2 ,3-dihydro-1,3-benzothiazol-2-ylidene]amino}pyridazin-3-yl)amino]1,3-thiazol-4-carboxylate.

To a solution of synthesis product 51 (100 mg, 0.16 mmol, 1 eq.) in dichloromethane (3 ml) was added ethyl isocyanate (0.02 ml, 0.2 mmol, 1.2 eq.) followed by the addition of triethylamine (0.03 ml, 0.25 mmol, 1.5 eq.) , and the mixture was stirred at ambient temperature overnight. The reaction mixture was partitioned between dichloromethane and saturated aqueous ammonium chloride, and the organic phase was washed with brine, dried (magnesium sulfate) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 4 g RediSep™ silica gel cartridge) eluting with a gradient of 0-100% ethyl acetate in isoheptane provided the title product as a cream solid (78 mg, 0.11 mmol, 70%) .

LC/MS (C ₃₁ H ₄₂ N ₈ O ₄ SiS ₂ ) 683 [M+H] ⁺ ; RT 1.38 (LCMS-V-BI).

1H NMR (400 MHz, DMSO-de) δ 7.86 (dd, J=7.6, 1.1 Hz, 1H), 7.69 (d, J=1.1 Hz, 1H), 7.51-7.39 (m, 2H), 7.29-7.20 ( m, 1H), 6.46 (t, J=5.6 Hz, 1H), 5.87 (s, 2H), 4.61-4.49 (m, 1H), 4.35-4.23 (m, 4H), 3.82 (dd, J=8.2, 6.0 Hz, 2H), 3.79 (s, 3H), 3.77-3.68 (m, 2H), 3.09-3.01 (m, 2H), 2.47 (d, J=1.0 Hz, 3H), 1.32 (t, J=7.1 Hz, 3H), 1.03 (t, J=7.1 Hz, 3H), 1.00-0.88 (m, 2H), -0.11 (s, 9H).

Step B: ethyl 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)-5-[1(ethylcarbamoyl)azetidine-3 - yl]-1,3-thiazol-4-carboxylate.

To a solution of the product from Step A (78 mg, 0.11 mmol, 1 eq.) in dichloromethane (3 mL) was added trifluoroacetic acid (0.4 mL, 5.37 mmol, 47 eq.) and the mixture was stirred at ambient temperature overnight. The reaction mixture was quenched by adding saturated aqueous sodium bicarbonate and extracted with dichloromethane, and the organic extract was washed with brine, dried (magnesium sulfate) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 4 g RediSep™ silica gel cartridge) eluting with a gradient of 0-10% methanol in dichloromethane gave the title product as a yellow glass (44 mg, 0.08 mmol, 70%) .

LC/MS (C ₂₅ H ₂₈ N ₈ O ₃ S ₂ ) 553 [M+H] ⁺ ; RT 1.00 (LCMS-V-BI).

1H NMR (400 MHz, DMSO-de) δ 11.18 (br s, 1H), 7.93 (br s, 1H), 7.70 (s, 1H), 7.46 (br s, 1H), 7.43-7.34

- 212 046336 (m, 1H), 7.27-7.14 (m, 1H), 6.46 (t, J=5.6 Hz, 1H), 4.62-4.48 (m, 1H), 4.36-4.22 (m, 4H), 3.87- 3.79 (m, 2H), 3.77 (s, 3H), 3.11-2.98 (m, 2H), 2.47 (s, 3H), 1.32 (t, J=7.1 Hz, 3H), 1.02 (t, 3H).

Step C: 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)-5-[1(ethylcarbamoyl)azetidin-3-yl ]-1,3-thiazole-4-carboxylic acid.

To a solution of the product from step B (44 mg, 0.08 mmol, 1 eq.) in 1,4-dioxane (3 ml) was added a 1 M aqueous solution of lithium hydroxide (0.4 ml, 0.4 mmol, 5 eq.), and the mixture was heated at 100°C for 4.5 h. Purification by reverse phase automated flash chromatography (CombiFlash Rf, C18 13 g RediSep column) eluting with a gradient of 5-95% acetonitrile in water gave the title product as a yellow solid ( 27.4 mg, 0.05 mmol, 66%) [as lithium salt].

HRMS-ERI (m/z) [M+H]+, calculated for C ₂₃ H ₂₅ N ₈ O ₃ S ₂ : 525.1486; found: 525.1516.

Example 161: 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)-5-[(2S)-3{4-[ 3-(dimethylamino)prop-1-in-1-yl]-2-fluorophenoxy}-2-methylpropyl]-1,3-thiazol-4-carboxylic acid

Step A: ethyl 5-[(2S)-3-{4-[3-(dimethylamino)prop-1-in-1-yl]-2-fluorophenoxy}-2-methylnropyl]-2[methyl(5-methyl -6-{[(2Z)-3-{[2-(trimethylsilyl)ethoxy]methyl}-2,3-dihydro-1,3-benzothiazol-2ylidene]amino}pyridazin-3-yl)amino]-1, 3-thiazole-4-carboxylate.

To a solution of synthesis product 3zf (40 mg, 0.09 mmol, 1 equiv.) in 1,4-dioxane (3 ml) was added synthesis product 4a (37.6 mg, 0.09 mmol, 1 equiv.), N-diisopropylethylamine (0.05 ml, 0.28 mmol, 3 eq.), cesium carbonate (90.2 mg, 0.28 mmol, 3 eq.) and xanthos (10.7 mg, 0.02 mmol, 0.2 eq.) followed by the addition of tris(dibenzylideneacetone)dipalladium(0) (8.45 mg, 0.01 mmol , 0.1 eq.), and the mixture was heated at 100°C for 24 h, and then allowed to cool to ambient temperature. The reaction mixture was partitioned between dichloromethane and water, and the organic phase was washed with brine, dried (magnesium sulfate) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 4 g RediSep™ silica gel cartridge) eluting with a gradient of 07% methanol in dichloromethane gave the title product as a brown glass (30.7 mg, 0.04 mmol, 41%).

LC/MS (C ₄₀ H ₅₀ FN ₇ O ₄ SiS ₂ ) 804 [M+H]+; RT 1.59 (LCMS-V-BI).

¹ H NMR (400 MHz, DMSOC,) δ 7.88-7.81 (m, 1H), 7.67 (d, J=1.1 Hz, 1H), 7.51-7.39 (m, 2H), 7.34-7.23 (m, 2H), 7.22-7.11 (m, 2H), 5.86 (s, 2H), 4.26 (q, J=7.1 Hz, 2H), 4.07-3.94 (m, 2H), 3.78 (s, 2H), 3.76-3.68 (m, 2H), 3.37 (s, 3H), 3.32-3.26 (m, 1H), 3.18-3.06 (m, 1H), 2.60-2.52 (m, 1H), 2.46 (d, J=1.0 Hz, ZN), 2.19 (s, 6H), 1.30 (t, J=7.1 Hz, 3H), 1.04 (d, J=6.7 Hz, 3H), 0.97 - 0.88 (m, 2H), -0.11 (s, 9H).

Step B: ethyl 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)-5-[^)3-{4-[ 3-(dimethylamino)prop-1-yn-1-yl]-2-fluorophenoxy}-2-methylpropyl]-1,3-thiazol-4-carboxylate.

To a solution of the product from step A (30.7 mg, 0.04 mmol, 1 eq.) in tetrahydrofuran (5 ml) was added tetrabutylammonium fluoride (1 M in tetrahydrofuran, 0.22 ml, 0.22 mmol, 6 eq.) and ethylenediamine (0.02 ml, 0.22 mmol , 6 eq.) in tetrahydrofuran (5 ml), and the mixture was refluxed overnight. The reaction mixture was partitioned between ethyl acetate and water, the aqueous phase was extracted with ethyl acetate (x2) and then dichloromethane (x2), and the combined organic extracts were dried (magnesium sulfate) and concentrated in vacuo to give the desired product, which was directly used in the next step without further purification.

Step C: 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylniridazin-3-yl}(methyl)amino)-5-[(2S)-3-{4[ 3-(dimethylamino)prop-1-yn-1-yl]-2-fluorophenoxy}-2-methylpropyl]-1,3-thiazol-4-carboxylic acid.

To a solution of the product from step B (38 mg, 0.06 mmol, 1 eq.) in 1,4-dioxane (3 ml) was added a 1 M aqueous solution of lithium hydroxide (0.56 ml, 0.56 mmol, 10 eq.), and the mixture was heated at 100°C for 2.5 h. The reaction mixture was allowed to cool to ambient temperature and purified by reverse phase flash chromatography (CombiFlash Rf, C18 13 g RediSep column) eluting with a gradient of 5-95% acetonitrile in water to give brown glass. The substance was dissolved in a mixture of dichloromethane and methanol, and applied to a 2 g PE-AX cartridge (pre-conditioned with dichloromethane, then methanol). The cartridge was washed with dichloromethane after

- 213 046336 washing with methanol, then eluted with 10% formic acid in dichloromethane, and the solvent was removed and dried in vacuo to give the title product as a brown glass (6.4 mg, 0.01 mmol, 18%) [as formic acid salt].

HRMS-ERI (m/z) [M+H]+, calculated for C ₃₂ H ₃₃ FN ₇ O ₃ S2: 646.2065; found: 646.2110.

Example 162: 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)-5-[(1E)-2methyl-3-phenoxyprop -1-en-1-yl]-1,3-thiazol-4-carboxylic acid

Step A: ethyl 2-[methyl(5-methyl-6-{[(2Z)-3-{[2-(trimethylsilyl)ethoxy]methyl}-2,3-dihydro-1,3benzothiazol-2-ylidene] amino } pyridazin-3 -yl)amino]-5-[(1 E)-2-methyl-3-phenoxyprop-1-en-1-yl]-1,3thiazol-4-carboxylate.

To a solution of synthesis product 5t (160 mg, 0.26 mmol, 1 eq.) in toluene (3 ml) was added phenol (34 μl, 0.38 mmol, 1.5 eq.), triphenylphosphine (100 mg, 0.38 mmol, 1.5 eq.) and diisopropyl azodicarboxylate (75 μl, 0.38 mmol, 1.5 eq.), and the mixture was heated at 120°C for 1 h under microwave irradiation. The reaction mixture was partitioned between dichloromethane and water, and the organic phase was separated and dried (PTFE phase separator), and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 4 g RediSep™ silica gel cartridge) eluting with a gradient of 0-60% ethyl acetate in isoheptane gave the title product as a yellow gum (100 mg, 0.14 mmol, 56%).

LC/MS (C ₃₅ H ₄₂ N ₆ O ₄ SiS ₂ ) 703 [M+H] ⁺ ; RT 3.12 (LCMS-VC).

¹ H NMR (400 MHz, DMSO) δ 7.86-7.81 (m, 1H), 7.72 (d, 1H), 7.51-7.40 (m, 2H), 7.36-7.21 (m, 4H), 7.06-7.00 (m, 2H), 6.99-6.93 (m, 1H), 5.87 (s, 2H), 4.68 (s, 2H), 4.26 (q, J=7.1 Hz, 1H), 3.81 (s, 3H), 3.76-3.68 (m , 2H), 2.46 (s, 3H), 2.06 (s, 3H), 1.28 (t, 3H), 0.97-0.88 (m, 2H), -0.11 (s, 9H).

Step B: ethyl 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)-5-[(1E)2-methyl-3 -phenoxyprop-1-en-1-yl]-1,3-thiazol-4-carboxylate.

To a solution of the product from step A (496 mg, 0.71 mmol, 1 eq.) in tetrahydrofuran (15 ml) was added ethylenediamine (141 μl, 2.12 mmol, 3 eq.) and tetrabutylammonium fluoride (1 M in tetrahydrofuran; 2.12 ml, 2.12 mmol , 3 eq.), and the mixture was heated at 70°C overnight. The reaction mixture was partitioned between ethyl acetate and water, and the organic phase was dried (magnesium sulfate) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 24 g RediSep™ silica gel cartridge) eluting with a gradient of 0-100% ethyl acetate in isoheptane provided the title product as a yellow solid (37 mg, 0.06 mmol, 9%) .

LC/MS (C29H28N6O3S2) 573 [M+H] ⁺ ; RT 2.65 (LCMS-VC).

1H NMR (400 MHz, DMSO) δ 7.91 (br s, 1H), 7.73 (s, 1H), 7.43-7.35 (m, 2H), 7.34-7.28 (m, 3H), 7.257.16 (m, 1H) , 7.06-7.00 (m, 2H), 6.99-6.93 (m, 1H), 4.67 (s, 2H), 4.26 (q, J=7.1 Hz, 2H), 3.80 (s, 3H), 2.47 (s, 3H ), 2.05 (s, 3H), 1.28 (t, J=7.1 Hz, 3H).

Step C: 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)-5-[(1E)-2methyl-3-phenoxyprop -1-en-1-yl]-1,3-thiazol-4-carboxylic acid.

To a solution of the product from step B (37 mg, 0.06 mmol, 1 eq.) in 1,4-dioxane (4 ml) was added lithium hydroxide monohydrate (27.1 mg, 0.65 mmol, 10 eq.), and the mixture was heated to 110°C during the night. The reaction mixture was allowed to cool to ambient temperature and purified using automated flash column chromatography (CombiFlash Rf, 4 g RediSep™ silica gel cartridge) eluting with a 0-10% 7N gradient. a methanol solution of ammonia in dichloromethane to obtain a dark yellow solid. The solid was triturated with acetonitrile, filtered and dried in vacuo to give the title product as a yellow solid (12.1 mg, 0.02 mmol, 34%).

HRMS-ERI (m/z) [M+H] ⁺ , calculated for C ₂₇ H ₂₅ N ₆ O ₃ S ₂ : 545.1424; found: 545.1426.

Example 163: 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)-5-[(1E)-3{4-[ 3 -(dimethylamino)prop-1 -in-1 -yl] -2-fluorophenoxy} -2-methylprop-1 -en-1 -yl] -1,3-thiazol-4-carboxylic acid

- 214 046336

Step A: ethyl 5-[(1E)-3-{4-[3-(dimethylamino)prop-1-in-1-yl]-2-fluorophenoxy}-2-methylprop-1-en1-yl]-2 -[methyl(5-methyl-6-{[(^)-3-{[2-(trimethylsilyl)ethoxy]methyl}-2,3-dihydro-1,3-benzothiazol-2ylidene]amino}pyridazin-3- yl)amino]-1,3-thiazole-4-carboxylate.

To a solution of synthesis product 5m (618 mg, 0.99 mmol, 1 eq.) in toluene (18 ml) was added synthesis product 6b (286 mg, 1.48 mmol, 1.5 eq.), triphenylphosphine (388 mg, 1.48 mmol, 1.5 eq.) and diisopropyl azodicarboxylate (291 μL, 1.48 mmol, 1.5 eq.), and the mixture was heated at 120°C for 1 h under microwave irradiation. The reaction mixture was partitioned between dichloromethane and water, and the organic phase was separated and dried (PTFE phase separator), and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 24 g RediSep™ silica gel cartridge) eluting with a gradient of 0-100% ethyl acetate in isoheptane gave the title product as a brown oil (406 mg, 0.51 mmol, 51%).

LC/MS (C ₄₀ H4 ₈ FN ₇ O ₄ SiS ₂ ) 802 [M+H]+; RT 2.68 (LCMS-VC).

¹ H NMR (400 MHz, DMSO-do) δ 7.86-7.80 (m, 1H), 7.70 (s, 1H), 7.50-7.39 (m, 2H), 7.38-7.21 (m, 5H), 5.86 (s, 2H), 4.78 (s, 2H), 4.25 (q, J=7.2 Hz, 2H), 3.81 (s, 3H), 3.76-3.67 (m, 2H), 3.41 (s, 2H), 2.46 (d, J =1.0 Hz, 3H), 2.22 (s, 6H), 2.05 (s, 3H), 1.28 (t, J=7.1 Hz, 3H), 0.97-0.85 (m, 2H), -0.11 (s, 9H).

Step B: ethyl 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)-5-[(1E)3-{4- [3-(dimethylamino)prop-1-in-1-yl]-2-fluorophenoxy}-2-methylprop-1-en-1-yl]-1,3-thiazol-4carboxylate.

To a solution of the product from step A (406 mg, 0.51 mmol, 1 eq.) in acetonitrile (5 ml) was added pyridinium polyfluoride (0.88 ml, 10.1 mmol, 20 eq.) and the mixture was heated at 60°C overnight. The reaction mixture was allowed to cool to ambient temperature, then partitioned between dichloromethane and 2N. aqueous sodium hydroxide solution (25 ml), and the organic phase was separated and dried (PTFE phase separator), and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 24 g RediSep™ silica gel cartridge) eluting with a gradient of 0-5% methanol in dichloromethane provided the title product as a yellow solid (139 mg, 0.21 mmol, 41%) .

LC/MS (C ₃₄ H ₃₄ FN ₇ O ₃ S ₂ ) 672 [M+H] ⁺ ; RT 2.07 (LCMS-VC).

1H NMR (400 MHz, DMSO^ ₆ ) δ 11.56 (br s, 1H), 7.90 (d, J=7.6 Hz, 1H), 7.72 (s, 1H), 7.57-7.50 (m, 1H), 7.44-7.28 (m, 3H), 7.27-7.17 (m, 3H), 4.79 (s, 2H), 4.27 (q, J=7.2 Hz, 2H), 3.81 (s, 3H), 3.43 (s, 2H), 2.47 ( s, 3H), 2.23 (s, 6H), 2.06 (s, 3H), 1.29 (t, J=7.1 Hz, 3H).

Step C: 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)-5-[(1E)-3-{4[ 3-(dimethylamino)prop-1-in-1-yl]-2-fluorophenoxy}-2-methylprop-1-en-1-yl]-1,3-thiazol-4-carboxylic acid.

To a solution of the product from step B (139 mg, 0.21 mmol, 1 eq.) in 1,4-dioxane (10 ml) was added lithium hydroxide monohydrate (86.8 mg, 2.07 mmol, 10 eq.), and the mixture was heated to 110°C for 4 hours. The reaction mixture was allowed to cool to ambient temperature and the solvent was removed by rotary evaporation. Purification by automated flash column chromatography (CombiFlash Rf, 12 g RediSep™ silica gel cartridge) eluting with a 0-25% 7N gradient. a methanol solution of ammonia in dichloromethane gave a solid. The material was triturated with acetonitrile, filtered and dried in vacuo to give the title product as a yellow solid (77.9 mg, 0.12 mmol, 59%).

MSHR-ERI (m/z) [M+H] ⁺ , calculated for C ₃₂ H ₃₁ FN ₇ O ₃ S ₂ : 644.1908; found: 644.1908.

Example 164: 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)-5-(3-{4-[3(dimethylamino )prop-1-yn-1-yl]-2-fluorophenoxy}-2-hydroxypropyl)-1,3-thiazol-4-carboxylic acid

- 215 046336

Step A: ethyl 5-(3-{4-[3-(dimethylamino)prop-1-yn-1-yl]-2-fluorophenoxy}-2-(oxan-2yloxy)propyl)-2-[methyl(5 -methyl-6-{ [(2Z)-3 - {[2-(trimethylsilyl)ethoxy]methyl }-2,3-dihydro-1,3benzothiazol-2-ylidene]amino}pyridazin-3-yl)amino]- 1,3-thiazole-4-carboxylate.

Synthesis product 4a (150 mg, 0.37 mmol, 1.16 eq.) and synthesis product 3zg (165 mg, 0.32 mmol, 1 eq.) were added to a solution of tris(dibenzylideneacetone)dipalladium(0) (29.1 mg, 0.03 mmol, 0.1 eq. ) and xantphos (36.8 mg, 0.06 mmol, 0.2 eq.) in 1,4-dioxane (8 ml). Cesium carbonate (310 mg, 0.95 mmol, 3 eq.) and N,N-diisopropylethylamine (0.2 mL, 0.95 mmol, 3 eq.) were added and the mixture was heated in a sealed tube at 125°C for 18 h. The suspension was allowed to cool and the mixture was filtered through celite, the solids were washed with ethyl acetate (30 ml) and the filtrate was concentrated in vacuo. Purification by flash column chromatography (20 g silica gel) eluting with a gradient of 0-2.5% methanol in dichloromethane gave the title product as a dark brown gum (230 mg, 0.26 mmol, 81%).

LC/MS (C ₄₄ H ₅₆ FN ₇ O ₆ SiS ₂ ) 890 [M+H]+; RT 1.59 (LCMS-VB!).

Step B: ethyl 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)-5-(3-{4[3-( dimethylamino)prop-1-yn-1-yl]-2-fluorophenoxy}-2-hydroxypropyl)-1,3-thiazol-4-carboxylate.

To a solution of the product from step A (225 mg, 0.25 mmol, 1 eq.) in chloroform (4.5 ml), cooled in ice water, trifluoroacetic acid (0.5 ml, 6.32 mmol, 25 eq.) was added and the mixture was stirred for 1 hour at 0°C and 18 hours at ambient temperature. Dichloromethane (50 ml) was added and the mixture was washed successively with saturated aqueous sodium bicarbonate (30 ml), water (2x30 ml) and brine (30 ml), dried (magnesium sulfate) and concentrated in vacuo. Purification by flash column chromatography (20 g silica gel) eluting with a gradient of 0-5% methanol in dichloromethane gave the title product as a dark yellow solid (98 mg, 0.15 mmol, 57%).

LC/MS (C ₃₃ H ₃₄ FN ₇ O ₄ S ₂ ) 676 [M+H]+; RT 1.15 (LCMS-VB!).

1H NMR (400 MHz, DMSO-0 ₆ ) δ 7.86-7.63 (m, 2H), 7.48-7.31 (m, 2H), 7.30-7.21 (m, 1H), 7.20-7.11 (m, 2H), 6.96- 6.85 (m, 1H), 4.46-4.31 (m, 3H), 4.16-4.01 (m, 2H), 3.92-3.76 (m, 3H), 3.64-3.53 (m, 1H), 3.51-3.44 (m, 1H ), 3.42 (s, 3H), 2.44 (s, 3H), 2.34 (s, 6H), 1.43 (t, J=7.1 Hz, 3H).

Step C: 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)-5-(3-{4-[3(dimethylamino )prop-1 -yn-1 -yl]-2-fluorophenoxy}-2-hydroxypropyl)-1,3-thiazol-4-carboxylic acid.

To a solution of the product from step B (90 mg, 0.13 mmol, 1 eq.) in a 1:3 mixture of 1,4-dioxane/water (2 ml) was added lithium hydroxide monohydrate (11.2 mg, 0.27 mmol, 2 eq.) and a suspension heated at 40°C for 7 hours. The reaction mixture was allowed to cool and the resulting solution was acidified with acetic acid to pH 5 to obtain a yellow precipitate. The solids were collected by filtration, washed with water and dried in vacuo to give the title product as a yellow solid (54 mg, 0.08 mmol, 63%).

HRMS-ERI (m/z) [M+H]+, calculated for C ₃₁ H ₃₁ FN ₇ O ₄ S ₂ : 648.1857; found: 648.1888.

Example 165: 2-[[6-(1,3-Benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]-[5-[methyl(4piperidyl)amino]pentyl]amino]-5-[3- [4-[3-(dimethylamino)prop-1-ynyl]-2-fluorophenoxy]propyl]thiazole4-carboxylic acid

- 216 046336

Step A: methyl 2-[5-[(1-tert-butoxycarbonyl-4-piperidyl)-methylamino]pentyl-[5-methyl-6-[^)[3-(2-trimethylsilylethoxymethyl)-1,3-benzothiazole -2-ylidene]amino]pyridazin-3-yl]amino]-5-[3-[4-[3(dimethylamino)prop-1-ynyl]-2-fluorophenoxy]propyl]thiazole-4-carboxylate.

Using a general alkylation procedure with an in situ tosylate starting from synthesis 5f and tert-butyl 4-(methylamino)piperidine-1-carboxylate as a suitable amine, the desired product was obtained.

HRMS-ERI (m/z): [M+2H] ²⁺ , calculated for C ₅₃ H ₇₄ FN9O ₆ S ₂ Si: 522.7551; found: 522.7542.

Step B: 2-[[6-(1,3-benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]-[5-[methyl(4piperidyl)amino]pentyl]amino]-5-[3- [4-[3-(dimethylamino)prop-1-ynyl]-2-fluorophenoxy]propyl]thiazole4-carboxylic acid.

Using the general deprotection and hydrolysis procedure followed by repurification by reverse phase preparative chromatography (C18, 0.1% TFA in water:MeCN), the TFA salt of the desired product was obtained from the product from step A.

MSHR-ERI (m/z): [M+2H]2 ⁺ , calculated for C ₄₁ H ₅₂ FN9O ₃ S ₂ : 400.6804; found: 400.6803.

Example 166: 2-[[6-(1,3-benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]-[5-(2morpholinoethylamino)pentyl]amino]-5-[3-[4-[ 3-(dimethylamino)prop-1-ynyl]-2fluorophenoxy]propyl]thiazole-4-carboxylic acid

Step A: methyl 5-[3-[4-[3-(dimethylamino)prop-1-ynyl]-2-fluorophenoxy]propyl]-2-[[5-methyl-6[^)-[3-(2 -trimethylsilylethoxymethyl)-1,3-benzothiazol-2-ylidene]amino]pyridazin-3-yl]-[5-(2morpholinoethylamino)pentyl]amino]thiazol-4-carboxylate.

Using a general alkylation procedure with an in situ formed tosylate starting from the synthesis product 5f and 2-morpholinoethanamine as a suitable amine, the desired product was obtained.

¹ H NMR (500 MHz, DMSO-06) δ ppm 7.84 (dm, 1H), 7.66 (s, 1H), 7.47 (dm, 1H), 7.44 (m, 1H), 7.30 (dd, 1H), 7.25 (m, 1H), 7.21 (dd, 1H), 7.16 (t, 1H), 5.86 (s, 2H), 4.37 (t, 2H), 4.14 (t, 2H), 3.78 (s, 3H), 3.72 (m, 2H), 3.52 (m, 4H), 3.38 ( s, 2H), 3.26 (m, 2H), 2.56 (t, 2H), 2.49 (t, 2H), 2.46 (s, 3H), 2.32 (t, 2H), 2.31 (m, 4H), 2.19 (s , 6H), 2.12 (m, 2H), 1.69 (m, 2H), 1.47 (m, 2H), 1.37 (m, 2H), 0.92 (m, 2H), -0.11 (s, 9H); MSHR-ERI (m/z): [M+2H] ²⁺ , calculated for C48H68FN9O5S2Si: 480.7269; found: 480.7264.

Step B: 2-[[6-(1,3-benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]-[5-(2morpholinoethylamino)pentyl]amino]-5-[3-[4-[ 3-(dimethylamino)prop-1-ynyl]-2fluorophenoxy]propyl]thiazole-4-carboxylic acid.

Using the general deprotection and hydrolysis procedure followed by repurification by reverse phase preparative chromatography (C18, 0.1% TFA in water:MeCN), the TFA salt of the desired product was obtained from the product from step A.

MSHR-ERI (m/z): [M+2H] ²⁺ , calculated for C ₄₁ H ₅₂ FN ₉ O ₄ S ₂ : 408.6779; found: 408.6769.

Example 167: 2-[[6-(1,3-Benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]-[5-[2-(4-methylpiperazin-1yl)ethylamino]pentyl]amino]- 5-[3-[4-[3-(dimethylamino)prop-1-ynyl]-2-fluorophenoxy]propyl]thiazole-4carboxylic acid

- 217 046336

Step A: methyl 5-[3-[4-[3-(dimethylamino)prop-1-ynyl]-2-fluorophenoxy]propyl]-2-[5-[2-(4methylpiperazin-1 -yl)ethylamino]pentyl -[5-methyl-6-[^)-[3-(2-trimethylsilylethoxymethyl)-1,3benzothiazol-2-ylidene]amino]pyridazin-3-yl]amino]thiazol-4-carboxylate.

Using a general alkylation procedure with an in situ formed tosylate starting from the synthesis product 5f and 2-(4-methylpiperazin-1-yl)ethanamine as a suitable amine, the desired product was obtained.

¹ H NMR (500 MHz, DMSO-dg) δ ppm 7.84 (dm, 1H), 7.67 (s, 1H), 7.47 (dm, 1H), 7.44 (m, 1H), 7.30 (dd, 1H), 7.26 (m, 1H), 7.21 (dd, 1H), 7.15 (t, 1H), 5.86 (s, 2H), 4.36 (t, 2H), 4.14 (t, 2H), 3.78 (s, 3H), 3.72 (m, 2H), 3.38 (s, 2H), 3.26 ( m, 2H), 2.53 (t, 2H), 2.50-2.10 (brs, 4H), 2.50-2.10 (brs, 4H), 2.48 (t, 2H), 2.46 (s, 3H), 2.30 (t, 2H) , 2.20 (s, 6H), 2.13 (m, 2H), 2.10 (s, 3H), 1.69 (m, 2H), 1.45 (m, 2H), 1.37 (m, 2H), 0.92 (m, 2H), -0.11 (s, 9H); HRMS-ERI (m/z): [M+2H] ²⁺ , calculated for C ₄₉ H ₇₁ FN ₁₀ O ₄ S ₂ Si: 487.2422; found: 487.2422.

Step B: 2-[[6-(1,3-benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]-[5-[2-(4-methylpiperazin-1yl)ethylamino]pentyl]amino]- 5-[3-[4-[3-(dimethylamino)prop-1-ynyl]-2-fluorophenoxy]propyl]thiazole-4carboxylic acid.

Using the general deprotection and hydrolysis procedure followed by repurification by reverse phase preparative chromatography (C18, 0.1% TFA in water:MeCN), the TFA salt of the desired product was obtained from the product from step A.

MSHR-ERI (m/z): [M+2H] ²⁺ , calculated for C ₄₂ H ₅₅ FN ₁₀ O ₃ S ₂ : 415.19366; found: 415.1939.

Example 168: 2-[[6-(1,3-Benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]-[4-[2-(4-methylpiperazin-1yl)ethylamino]butyl]amino]- 5-[3-[4-[3-(dimethylamino)prop-1-ynyl]-2-fluorophenoxy]propyl]thiazole-4carboxylic acid

Step A: methyl 5-[3-[4-[3-(dimethylamino)prop-1-ynyl]-2-fluorophenoxy]propyl]-2-[4-[2-(4methylpiperazin-1-yl)ethylamino]butyl -[5-methyl-6-[^)-[3-(2-trimethylsilylethoxymethyl)-1,3benzothiazol-2-ylidene]amino]pyridazin-3-yl]amino]thiazol-4-carboxylate.

Using the general alkylation procedure with an in situ formed tosylate starting from the synthesis product 5d and 2-(4-methylpiperazin-1-yl)ethanamine as a suitable amine, the desired product was obtained.

¹ H NMR (500 MHz, DMSO-dg) δ ppm 2.50-2.00 (brs, 8H), 7.84 (dm, 1H), 7.72 (s, 1H), 7.47 (dm, 1H), 7.43 (m, 1H), 7.30 (dd, 1H), 7.25 (m, 1H) , 7.21 (dd, 1H), 7.16 (t, 1H), 5.86 (s, 2H), 4.38 (t, 2H), 4.15 (t, 2H), 3.78 (s, 3H), 3.72 (m, 2H), 3.39 (s, 2H), 3.27 (t, 2H), 2.56 (t, 2H), 2.54 (t, 2H), 2.46 (s, 3H), 2.31 (t, 2H), 2.20 (brs, 6H), 2.13 (m, 2H), 2.06 (s, 3H), 1.71 (m, 2H), 1.47 (m, 2H), 0.92 (m, 2H), -0.11 (s, 9H); MSHR-ERI (m/z): [M+2H] ²⁺ , calculated for C ₄₈ H ₆₉ FN ₁₀ O ₄ S ₂ Si: 480.2344; found: 480.2340.

Step B: 2-[[6-(1,3-benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]-[4-[2-(4-methylpiperazin-1yl)ethylamino]butyl]amino]- 5-[3-[4-[3-(dimethylamino)prop-1-ynyl]-2-fluorophenoxy]propyl]thiazole-4carboxylic acid.

Using a general deprotection and hydrolysis procedure followed by repurification by reverse phase preparative chromatography (C18, 0.1% TFA in water:MeCN), based on

- 218 046336 from the product from stage A, the TFA salt of the target product was obtained.

MSHR-ERI (m/z): [M+2H]2+, calculated for C ₄₁ H ₅₃ FN ₁₀ O ₃ S ₂ : 408.1858; found: 408.1857.

Example 169: 2-[[6-(1,3-Benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]-[4-hydroxy-5-[methyl-[2(methylamino)ethyl] amino] pentyl ]amino]-5-[3-[4-[3-(dimethylamino)prop-1-inyl]-2fluorophenoxy]propyl]thiazole-4-carboxylic acid

Step A: methyl 2-[[5-[2-[tert-butoxycarbonyl(methyl)amino]ethylmethylamino]-4-[tert-butyl(diphenyl)silyl]oxypentyl]-[5-methyl-6-[(2)-[ 3-(2-trimethylsilylethoxymethyl)-1,3-benzothiazol-2ylidene]amino]pyridazin-3-yl]amino]-5-[3-[4-[3-(dimethylamino)prop-1-ynyl]-2fluorophenoxy] propyl]thiazole-4-carboxylate.

Using the general tosylate alkylation procedure, starting from the synthesis product 5b and tert-butyl Nmethyl-N-[2-(methylamino)ethyl]carbamate as a suitable amine, the desired product was obtained.

Step B: 2-[[6-(1,3-Benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]-[4-hydroxy-5-[methyl-[2(methylamino)ethyl] amino] pentyl ] amino]-5-[3-[4-[3 -(dimethylamino)prop-1-inyl]-2fluorophenoxy]propyl]thiazole-4-carboxylic acid.

Using a general deprotection and hydrolysis procedure, starting with the product from Step A as the appropriate methyl ester, the desired product was obtained.

MSHR-ERI (m/z): [M+H]+, calculated for C ₃₉ H4 ₉ FN ₉ O ₄ S ₂ : 790.3327; found: 790.3317.

Example 170: 2-[[6-(1,3-Benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]-[5-[2(diethylamino)ethylamino]-4-hydroxypentyl]amino]-5- [3-[4-[3 -(dimethylamino)prop-1-ynyl]-2fluorophenoxy]propyl]thiazole-4-carboxylic acid

ABOUT

Step A: methyl 2-[[4-[tert-butyl(diphenyl)silyl]oxy-5-[2-(diethylamino)ethylamino]pentyl]-[5methyl-6-[(2)-[3-(2- trimethylsilylethoxymethyl)-1,3-benzothiazol-2-ylidene]amino]pyridazin-3-yl]amino]5-[3-[4-[3-(dimethylamino)prop-1-ynyl]-2-fluorophenoxy]propyl] thiazole-4-carboxylate.

Using the general tosylate alkylation procedure from synthesis 5b and N',N'diethylethane-1,2-diamine as a suitable amine, the desired product was obtained.

Step B: 2-[[6-(1,3-benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]-[5-[2(diethylamino)ethylamino]-4-hydroxypentyl]amino]-5- [3-[4-[3-(dimethylamino)prop-1-ynyl]-2fluorophenoxy]propyl]thiazole-4-carboxylic acid.

Using a general deprotection and hydrolysis procedure, starting with the product from Step A as the appropriate methyl ester, the desired product was obtained.

MSHR-ERI (m/z): [M+H]+, calculated for C ₄₁ H ₅₃ FN ₉ O ₄ S ₂ : 818.3640; found: 818.3613.

Example 171: 2-[[5-(4-Amino-1-piperidyl)-4-hydroxypentyl]-[6-(1,3-benzothiazol-2-ylamino)-5methylpyridazin-3-yl]amino]-5- [3-[4-[3-(dimethylamino)prop-1-ynyl]-2-fluorophenoxy]propyl]thiazole-4carboxylic acid

- 219 046336

Step A: methyl 2-[[5-[4-(tert-butoxycarbonylamino)-1-piperidyl]-4-[tert-butyl(diphenyl)silyl]oxypentyl]-[5-methyl-6-[^)-[3- (2-trimethylsilylethoxymethyl)-1,3-benzothiazol-2ylidene]amino]pyridazin-3-yl]amino]-5-[3-[4-[3-(dimethylamino)prop-1-ynyl]-2fluorophenoxy]propyl] thiazole-4-carboxylate.

Using the general tosylate alkylation procedure, starting from synthesis 5b and tert-butyl X-(4-piperidyl)carbamate as a suitable amine, the desired product was obtained.

Step B: 2-[[5-(4-amino-1-piperidyl)-4-hydroxypentyl]-[6-(1,3-benzothiazol-2-ylamino)-5methylpyridazin-3-yl]amino]-5- [3-[4-[3-(dimethylamino)prop-1-ynyl]-2-fluorophenoxy]propyl]thiazole-4carboxylic acid.

Using a general deprotection and hydrolysis procedure, starting with the product from Step A as the appropriate methyl ester, the desired product was obtained.

MSHR-ERI (m/z): [M+H]+, calculated for C ₄₀ H ₄₉ FN9O ₄ S ₂ : 802.3327; found: 802.3324.

Example 172: 2-[[6-(1,3-benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]-(4-hydroxy-5-piperazin1-ylpentyl)amino]-5-[3-[ 4-[3-(dimethylamino)prop-1-ynyl]-2-fluorophenoxy]propyl]thiazole-4-carboxylic acid

Step A: methyl 2-[[5-(4-tert-butoxycarbonylpiperazin-1-yl)-4-[tert-butyl(diphenyl)silyl]oxypentyl]-[5-methyl-6-[^)-[3-(2 -trimethylsilylethoxymethyl)-1,3-benzothiazol-2ylidene]amino]pyridazin-3-yl]amino]-5-[3-[4-[3-(dimethylamino)prop-1-ynyl]-2fluorophenoxy]propyl]thiazol- 4-carboxylate.

Using the general tosylate alkylation procedure, starting from synthesis 5b and tert-butyl piperazine-1-carboxylate as a suitable amine, the desired product was obtained.

Step B: 2-[[6-(1,3-benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]-(4-hydroxy-5-piperazin-1ylpentyl)amino]-5-[3-[ 4-[3-(dimethylamino)prop-1-ynyl]-2-fluorophenoxy]propyl]thiazole-4-carboxylic acid.

Using a general deprotection and hydrolysis procedure, starting with the product from Step A as the appropriate methyl ester, the desired product was obtained.

MSHR-ERI (m/z): [M+2H]2 ⁺ , calculated for C ₃₉ H ₄₈ FN ₉ O ₄ S ₂ : 394.6622; found: 394.6614.

Example 173: 2-[[6-(1,3-benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]-[4-hydroxy-5-[2-(1piperidyl)ethylamino]pentyl]amino]- 5-[3-[4-[3-(dimethylamino)prop-1-ynyl]-2fluorophenoxy]propyl]thiazole-4-carboxylic acid

- 220 046336

Step A: methyl 2-[[4-[tert-butyl(diphenyl)silyl]oxy-5-[2-(1-piperidyl)ethylamino]pentyl]-[5methyl-6-[^)-[3-(2 -trimethylsilylethoxymethyl)-1,3-benzothiazol-2-ylidene]amino]pyridazin-3-yl]amino]5-[3-[4-[3-(dimethylamino)prop-1-ynyl]-2-fluorophenoxy]propyl ]thiazole-4-carboxylate.

Using the general tosylate alkylation procedure, starting from the synthesis product 5b and 2-(1piperidyl)ethanamine as a suitable amine, the desired product was obtained.

Step B: 2-[[6-(1,3-benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]-[4-hydroxy-5-[2-(1piperidyl)ethylamino]pentyl]amino]- 5-[3-[4-[3-(dimethylamino)prop-1-ynyl]-2-fluorophenoxy]propyl]thiazole-4-carboxylic acid.

Using a general deprotection and hydrolysis procedure, starting with the product from Step A as the appropriate methyl ester, the desired product was obtained.

MSHR-ERI (m/z): [M+2H] ²⁺ , calculated for C ₄₂ H ₅₄ FN9O ₄ S ₂ : 415.6857; found: 415.6853.

Example 174: 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)-5-[(1E)-3{ 2-fluoro -4-[3-(methylamino)prop-1-yn-1-yl]phenoxy}-2-methylprop-1-en-1-yl]-1,3-thiazol-4-carboxylic acid

Step A: ethyl 5-[(1E)-3-[4-(3-{[(tert-butoxy)carbonyl](methyl)amino}prop-1-yn-1-yl)-2fluorophenoxy]-2-methylprop -1-en-1-yl]-2-[methyl(5-methyl-6- {[(2Z)-3-{ [2-(trimethylsilyl)ethoxy] methyl } 2,3-dihydro-1,3- benzothiazol-2-ylidene] amino } pyridazin-3-yl) amino] -1,3-thiazol-4-carboxylate.

To a solution of synthesis product 5m (641 mg, 1.02 mmol, 1 eq.) in toluene (18 ml) was added synthesis product 6a (428 mg, 1.53 mmol, 1.5 eq.), triphenylphosphine (402 mg, 1.53 mmol, 1.5 eq.) and diisopropyl azodicarboxylate (0.3 ml, 1.53 mmol, 1.5 eq.), and the mixture was heated at 120°C for 1 hour. The reaction mixture was partitioned between dichloromethane and water, and the organic phase was separated and dried (PTFE phase separator), and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 24 g RediSep™ silica gel cartridge) eluting with a gradient of 0-40% ethyl acetate in isoheptane gave the title product as a yellow solid (380 mg, 0.43 mmol, 42%).

LC/MS (C ₄₄ H ₅₄ FN ₇ O ₆ SiS ₂ ) 888 [M+H] ⁺ ; RT 1.43 (ZHMS-U-V2).

1H NMR (400 MHz, DMSO-d^δ 7.86-7.81 (m, 1H), 7.71 (d, 1H), 7.51-7.40 (m, 2H), 7.39-7.32 (m, 1H), 7.32-7.21 (m, 4H), 5.86 (s, 2H), 4.79 (s, 2H), 4.32-4.17 (m, 4H), 3.81 (s, 3H), 3.78-3.66 (m, 2H), 2.86 (s, 3H ), 2.46 (s, 3H), 2.06 (m, 3H), 1.41 (s, 9H), 1.28 (t, 3H), 0.97-0.88 (m, 2H), -0.11 (s, 9H).

Step B: ethyl 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)-5-[(1E)3-{2- fluoro-4-[3-(methylamino)prop-1-yn-1-yl]phenoxy}-2-methylprop-1-en-1-yl]-1,3-thiazol-4-carboxylate.

To a solution of the product from step A (380 mg, 0.43 mmol, 1 eq.) in acetonitrile (5 ml) was added pyridinium polyhydrogen fluoride (744 μl, 8.56 mmol, 20 eq.) and the mixture was heated at 60°C for 3 h The reaction mixture was allowed to cool to ambient temperature, then partitioned between dichloromethane and 2N. aqueous sodium hydroxide solution, the aqueous phase was extracted with a 1:3 mixture of isopropanol/dichloromethane, and the combined organics were dried (PTFE phase separator) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 12 g RediSep™ silica gel cartridge) eluting with a gradient of 0-6% methanol in dichloromethane provided the title product as a yellow solid (133 mg, 0.2 mmol, 47%) .

LC/MS (C ₃₃ H ₃₂ FN ₇ O ₃ S ₂ ) 658 [M+H] ⁺ ; RT 2.02 (LCMS-VC).

1H NMR (400 MHz, DMSO^) δ 7.90 (d, J=7.8 Hz, 1H), 7.73 (s, 1H), 7.54 (d, J=8.0 Hz, 1H), 7.44-7.36 (m, 1H), 7.35-7.28 (m, 2H), 7.27-7.18 (m, 3H), 4.79 (s, 2H), 4.27 (q, J=7.1 Hz, 2H), 3.81 (s, 3H), 3.49 (s, 2H) , 2.48 (d, J=1.0 Hz, 3H), 2.34 (s, 3H), 2.07 (s, 3H), 1.29 (t, 3H).

Step C: 2-({6-[( 1,3 -benzothiazol-2-yl)amino]-5 -methylpyridazin-3 -yl}(methyl)amino)-5-[( 1 E)-3-{2fluoro -4-[3-(methylamino)prop-1-yn-1-yl]phenoxy}-2-methylprop-1-en-1-yl]-1,3-thiazol-4-carboxylic acid.

To a solution of the product from step B (133 mg, 0.2 mmol, 1 eq.) in 1,4-dioxane (6 ml) was added lithium hydroxide monohydrate (84.9 mg, 2.02 mmol, 10 eq.), and the mixture was heated to 110°C for 4 hours. The reaction mixture was allowed to cool to ambient temperature and purified using automated flash column chromatography (CombiFlash Rf, 12 g RediSep™ silica gel cartridge) eluting with a gradient of 0-25% 7N. a methanol solution of ammonia in dichloromethane to obtain a solid. The solid was triturated with acetonitrile, filtered and dried in vacuum

- 221 046336 to obtain the target product in the form of a yellow solid (4.32 mg, 0.01 mmol, 3%).

MSHR-ERI (m/z): [M+H]+, calculated for C ₃₁ H ₂₉ FN7O3S ₂ : 630.1752; found: 630.1781.

Example 175: 2-[4-(4-Amino-1-piperidyl)butyl-[6-(1,3-benzothiazol-2-ylamino)-5-methylpyridazin3-yl]amino]-5-[3-[4 -[3-(dimethylamino)prop-1-ynyl]-2-fluorophenoxy]propyl]thiazole-4-carboxylic acid

Step A: methyl 2-[4-[4-(tert-butoxycarbonylamino)-1-piperidyl]butyl-[5-methyl-6-[^)-[3-(2trimethylsilylethoxymethyl)-1,3-benzothiazol-2- ylidene]amino]pyridazin-3-yl]amino]-5-[3-[4-[3(dimethylamino)prop-1-ynyl]-2-fluorophenoxy]propyl]thiazole-4-carboxylate.

Using the general alkylation procedure with the in situ formed tosylate, starting from the synthesis product 5d and tert-butyl Na(4-piperidyl)carbamate as a suitable amine, the desired product was obtained.

¹ H NMR (500 MHz, DMSO-de) δ ppm 7.65 (q, 1H), 7.33 (dd, 1H), 7.26 (dd, 1H), 7.18 (t, 1H), 6.80 (brs, 1H), 5.86 (s, 2H), 4.40 (t, 2H), 4.20 (t, 2H), 3.82 (s, 3H), 3.79 (s, 2H), 3.77 (m, 2H), 3.54 (brs, 1H), 3.36/3.00 (brs, 4H), 3.29 (t, 2H), 3.14 (t, 2H), 2.52 (brs, 6H), 2.48 (d, 3H), 2.15 (m, 2H), 1.94/1.66 (m+m, 4H), 1.79 (m, 2H), 1.76 (m, 2H), 1.40 (s, 9H), 0.94 (m, 2H), -0.07 (s, 9H); MSHR-ERI (m/z): [M+H]+, calculated for C ₅₁ H ₇₁ FN ₉ O ₆ S ₂ Si: 1016.4716; found: 1016.4704.

Step B: 2-[4-(4-amino-1-piperidyl)butyl-[6-(1,3-benzothiazol-2-ylamino)-5-methylpyridazin-3yl]amino]-5-[3-[4 -[3-(dimethylamino)prop-1-ynyl]-2-fluorophenoxy]propyl]thiazole-4-carboxylic acid.

Using the general deprotection and hydrolysis procedure followed by repurification by reverse phase preparative chromatography (C18, 0.1% TFA in water:MeCN), the TFA salt of the desired product was obtained from the product from step A.

MSHR-ERI (m/z): [M+H]+, calculated for C ₃₉ H ₄₆ FN ₉ O ₃ S ₂ : 772.3222; found: 772.3228.

Example 176: 2-[[6-(1,3-Benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]-[4-[2(diethylamino)ethylamino]butyl]amino]-5-[3- [4-[3-(dimethylamino)prop-1-ynyl]-2fluorophenoxy]propyl]thiazole-4-carboxylic acid

Step A: methyl 2-[4-[2-(diethylamino)ethylamino]butyl-[5-methyl-6-[^)-[3-(2trimethylsilylethoxymethyl)-1,3-benzothiazol-2-ylidene]amino]pyridazine -3-yl]amino]-5-[3-[4-[3(dimethylamino)prop-1-ynyl]-2-fluorophenoxy]propyl]thiazole-4-carboxylate.

Using a general alkylation procedure with an in situ formed tosylate, starting from the synthesis product 5d and N,N-diethylethane-1,2-diamine as a suitable amine, the desired product was obtained.

1H NMR (500 MHz, DMSO-d^ δ ppm 7.84 (dm, 1H), 7.71 (s, 1H), 7.47 (dm, 1H), 7.44 (m, 1H), 7.30 (dd, 1H), 7.25 (m, 1H), 7.21 (dd, 1H), 7.16 (t, 1H), 5.86 (s, 2H), 4.39 (t, 2H), 4.15 (t, 2H), 3.78 (s, 3H), 3.72 (m, 2H), 3.38 (s, 2H), 3.27 (t, 2H), 2.57 (t, 2H), 2.51 (t, 2H), 2.46 (s, 3H), 2.41 (t, 2H), 2.40 ( q, 4H), 2.19 (s, 6H), 2.12 (m, 2H), 1.71 (m, 2H), 1.48 (m, 2H), 0.92 (m, 2H), 0.89 (t, 6H), -0.11 ( s, 9H); HRMS-ERI (m/z): [M+H]+, calculated for C ₄₇ H ₆₇ FN ₉ O ₄ S ₂ Si: 932.4505; found: 932.4489.

Step B: 2-[[6-(1,3-benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]-[4-[2(diethylamino)ethylamino]butyl]amino]-5-[3- [4-[3-(dimethylamino)prop-1-ynyl]-2fluorophenoxy]propyl]thiazole-4-carboxylic acid.

Using the general technique of deprotection and hydrolysis followed by repurification from to

- 222 046336 using the power of reverse-phase preparative chromatography (C18, 0.1% TFA in a mixture of eoga:MeCN), starting from the product from stage A, the TFA salt of the target product was obtained.

MSHR-ERI (m/z): [M+2H]2+, calculated for C ₄₀ H ₅₂ FN9O ₃ S ₂ : 394.6804; found: 394.6802.

Example 177: 2-[5-(4-Amino-1-piperidyl)pentyl-[6-(1,3-benzothiazol-2-ylamino)-5methylpyridazin-3-yl]amino]-5-[3-[4 -[3-(dimethylamino)prop-1-ynyl]-2-fluorophenoxy]propyl]thiazole-4carboxylic acid

Step A: methyl 2-[5-[4-(tert-butoxycarbonylamino)-1-piperidyl]pentyl-[5-methyl-6-[(2)-[3-(2trimethylsilylethoxymethyl)-1,3-benzothiazol-2 -ylidene]amino]pyridazin-3-yl]amino]-5-[3-[4-[3(dimethylamino)prop-1-ynyl]-2-fluorophenoxy]propyl]thiazole-4-carboxylate.

Using the general alkylation procedure with the in situ formed tosylate, starting from the synthesis product 5f and tert-butyl Na(4-piperidyl)carbamate as a suitable amine, the desired product was obtained.

¹ H NMR (500 MHz, DMSO-de) δ ppm 7.91-7.18 (m, 4H), 7.70 (s, 1H), 7.33 (dd, 1H), 7.24 (dd, 1H), 7.17 (t, 1H), 7.04 (d, 1H), 5.87 (s, 2H) , 4.16 (t, 2H), 3.79 (s, 3H), 3.72 (t, 2H), 3.53 (s, 2H), 3.49/3.31 (m+m, 4H), 3.27 (t, 2H), 2.94 (t , 2H), 2.91 (m, 1H), 2.47 (s, 3H), 2.30 (s, 6H), 2.13 (m, 2H), 1.89/1.60 (m+m, 4H), 1.73 (m, 2H), 1.70 (m, 2H), 1.38 (m, 2H), 1.38 (m, 2H), 1.38 (s, 9H), 0.92 (t, 2H), -0.11 (s, 9H); HRMS-ERI (m/z): [M+2H] ²⁺ , calculated for C ₅₂ H ₇₄ FN ₉ O ₆ S ₂ Si: 515.7473; found: 515.74725.

Step B: 2-[5-(4-amino-1-piperidyl)pentyl-[6-(1,3-benzothiazol-2-ylamino)-5-methylpyridazin-3yl]amino]-5-[3-[4 -[3-(dimethylamino)prop-1-ynyl]-2-fluorophenoxy]propyl]thiazole-4-carboxylic acid.

Using the general deprotection and hydrolysis procedure followed by repurification by reverse phase preparative chromatography (C18, 0.1% TFA in water:MeCN), the TFA salt of the desired product was obtained from the product from step A.

MSHR-ERI (m/z): [M+2H] ²⁺ , calculated for C ₄₀ H ₅₀ FN ₉ O ₃ S ₂ : 393.6726; found: 393.6723.

Example 178: 2-[[6-(1,3-Benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]-[4-[methyl-[2(methylamino)ethyl] amino]butyl] amino] - 5-[3-[4-[3-(dimethylamino)prop-1-ynyl]-2fluorophenoxy]propyl]thiazole-4-carboxylic acid

Step A: methyl 2-[4-[2-[tert-butoxycarbonyl(methyl)amino]ethylmethylamino]butyl-[5-methyl-6[(2)-[3-(2-trimethylsilylethoxymethyl)-1,3-benzothiazole -2-ylidene]amino]pyridazin-3-yl]amino]-5-[3-[4[3-(dimethylamino)prop-1-ynyl]-2-fluorophenoxy]propyl]thiazole-4-carboxylate.

Using the general alkylation procedure with the in situ formed tosylate, starting from the synthesis product 5d and tert-butyl Nmethyl-N-[2-(methylamino)ethyl]carbamate as a suitable amine, the desired product was obtained.

¹ H NMR (500 MHz, DMSO-06) δ ppm 7.84 (dm, 1H), 7.68 (s, 1H), 7.46 (dm, 1H), 7.44 (m, 1H), 7.30 (dd, 1H), 7.25 (m, 1H), 7.20 (dd, 1H), 7.15 (t, 1H), 5.86 (s, 2H), 4.38 (t, 2H), 4.14 (t, 2H), 3.77 (s, 3h), 3.72 (m, 2H), 3.38 (s, 2H), 3.26 ( t, 2H), 3.19 (brm, 2H), 2.71 (brs, 3H), 2.46 (s, 3H), 2.37 (t, 2H), 2.36 (t, 2H), 2.19 (s, 6H), 2.15 (s , 3H), 2.12 (m, 2H), 1.69 (m, 2H), 1.48 (m, 2H), 1.31 (brs, 9H), 0.92 (m, 2H), -0.11 (s, 9H); MSVR-ERI

- 223 046336 (m/z): [M+H]+, calculated for C50H71FN9O6S2S1: 1004.4716; found: 502.73931.

Step B: 2-[[6-(1,3-benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]-[4-[methyl-[2(methylamino)ethyl] amino]butyl] amino] - 5-[3-[4-[3-(dimethylamino)prop-1-ynyl]-2fluorophenoxy]propyl]thiazole-4-carboxylic acid.

Using the general deprotection and hydrolysis procedure followed by repurification by reverse phase preparative chromatography (C18, 0.1% TFA in water:MeCN), the TFA salt of the desired product was obtained from the product from step A.

MSHR-ERI (m/z): [M+2H] ²⁺ , calculated for C ₃₈ H ₄₈ FN ₉ O ₃ S ₂ : 380.6647; found: 380.6640.

Example 179: 2-[[6-(1,3-Benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]-[4-[2-(1piperidyl)ethylamino]butyl]amino]-5-[3 -[4-[3-(dimethylamino)prop-1-ynyl]-2fluorophenoxy]propyl]thiazole-4-carboxylic acid

Step A: methyl 5-[3-[4-[3-(dimethylamino)prop-1-ynyl]-2-fluorophenoxy]propyl]-2-[[5-methyl-6[(2)-[3-( 2-trimethylsilylethoxymethyl)-1,3-benzothiazol-2-ylidene]amino]pyridazin-3-yl]-[4-[2-(1piperidyl)ethylamino]butyl]amino]thiazole-4-carboxylate.

Using a general alkylation procedure with an in situ formed tosylate starting from the synthesis product 5d and 2-(1-piperidyl)ethanamine as a suitable amine, the desired product was obtained.

1H NMR (500 MHz, DMSO-06) δ ppm 7.79 (dm, 1H), 7.68 (s, 1H), 7.48 (dm, 1H), 7.44 (m, 1H), 7.30 (dd, 1H), 7.26 (m, 1H), 7.24 (dd, 1H), 7.17 (t, 1H), 5.86 (s, 2H), 4.42 (t, 2H), 4.19 (t, 2H), 3.82 (s, 3H), 3.77 (m, 2H), 3.63 (brs, 2H), 3.28 ( t, 2H), 3.07 (t, 2H), 3.04 (t, 2H), 2.76 (t, 2H), 2.60 (brm, 4H), 2.48 (s, 3H), 2.41 (brs, 6H), 2.14 (m , 2H), 1.83 (m, 2h), 1.73 (m, 2H), 1.58 (m, 4H), 1.43 (m, 2H), 0.94 (m, 2H), -0.07 (s, 9H); MSHR-ERI (m/z): [M+H]+, calculated for C ₄₈ H ₆₇ FN ₉ O ₄ S ₂ Si: 944.4505; found: 944.44978.

Step B: 2-[[6-(1,3-benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]-[4-[2-(1piperidyl)ethylamino]butyl]amino]-5-[3 -[4-[3-(dimethylamino)prop-1-ynyl]-2fluorophenoxy]propyl]thiazole-4-carboxylic acid.

Using the general deprotection and hydrolysis procedure followed by repurification by reverse phase preparative chromatography (C18, 0.1% TFA in water:MeCN), the TFA salt of the desired product was obtained from the product from step A.

MSHR-ERI (m/z): [M+H]+, calculated for C ₄₁ H ₅₁ FN ₉ O ₃ S ₂ : 800.3535; found: 800.3540.

Example 180: 2-[[6-(1,3-benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]-[4-(2morpholinoethylamino)butyl]amino]-5-[3-[4-[ 3-(dimethylamino)prop-1-ynyl]-2fluorophenoxy]propyl]thiazole-4-carboxylic acid

Step A: methyl 5-[3-[4-[3-(dimethylamino)prop-1-ynyl]-2-fluorophenoxy]propyl]-2-[[5-methyl-6[(2)-[3-( 2-trimethylsilylethoxymethyl)-1,3-benzothiazol-2-ylidene]amino]pyridazin-3-yl]-[4-(2morpholinoethylamino)butyl]amino]thiazol-4-carboxylate.

Using a general alkylation procedure with an in situ formed tosylate starting from the synthesis product 5d and 2-morpholinoethanamine as a suitable amine, the desired product was obtained.

1H NMR (500 MHz, DMSO-06) δ ppm 7.81 (dm, 1H), 7.76 (s, 1H), 7.48 (dm, 1H), 7.44 (m, 1H), 7.40 (dd, 1H), 7.28 (dm, 1H), 7.27 (m, 1H), 7.20 (t, 1H), 5.87 (s, 2H), 4.42 (t, 2H), 4.17 (t, 2h), 3.85 (brs, 2H), 3.79 (s, 3H), 3.72 (m, 2H), 3.56 ( m, 4H), 3.27 (t, 2H), 3.01 (t, 2H), 2.99 (t, 2H), 2.58 (brm, 2H), 2.51 (brs, 6H), 2.47 (s,

- 224 046336

3H), 2.39 (brm, 4H), 2.13 (m, 2H), 1.84-1.68 (m, 4H), 0.92 (m, 2H), -0.11 (s, 9H); HRMS-ERI (m/z): [M+2H]2+, calculated for C ₄₇ H ₆₆ FN ₉ O5S ₂ Si: 473.7186; found: 473.7183.

Step B: 2-[[6-(1,3-benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]-[4-(2morpholinoethylamino)butyl]amino]-5-[3-[4-[ 3-(dimethylamino)prop-1-ynyl]-2fluorophenoxy]propyl]thiazole-4-carboxylic acid.

Using the general deprotection and hydrolysis procedure followed by repurification by reverse phase preparative chromatography (C18, 0.1% TFA in water:MeCN), the TFA salt of the desired product was obtained from the product from step A.

MSHR-ERI (m/z): [M+2H] ²⁺ , calculated for C ₄₀ H ₅₀ FN ₉ O ₄ S ₂ : 401.6700; found: 401.6697.

Example 181: 2-[[6-(1,3-Benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]-[4-[(1-methyl-4piperidyl)amino]butyl]amino]-5- [3-[4-[3-(dimethylamino)prop-1-ynyl]-2-fluorophenoxy]propyl]thiazole4-carboxylic acid

HN

Step A: methyl 5-[3-[4-[3-(dimethylamino)prop-1-ynyl]-2-fluorophenoxy]propyl]-2-[4-[(1-methyl-4piperidyl)amino]butyl-[ 5-methyl-6-[^)-[3-(2-trimethylsilylethoxymethyl)-1,3-benzothiazol-2ylidene]amino]pyridazin-3-yl]amino]thiazol-4-carboxylate.

Using a general alkylation procedure with an in situ formed tosylate starting from the synthesis product 5d and 1-methylpiperidin-4-amine as a suitable amine, the desired product was obtained.

¹ H NMR (500 MHz, DMSO-06) δ ppm 7.84 (dm, 1H), 7.70 (s, 1H), 7.47 (dm, 1H), 7.43 (m, 1H), 7.30 (dd, 1H), 7.25 (m, 1H), 7.21 (dm, 1H), 7.15 (t, 1H), 5.86 (s, 2H), 4.39 (t, 2H), 4.14 (t, 2H), 3.77 (s, 3H), 3.72 (m, 2H), 3.38 (s, 2H), 3.26 ( t, 2H), 2.65/1.84 (m+m, 4H), 2.56 (t, 2H), 2.46 (s, 3H), 2.27 (m, 1H), 2.19 (s, 6H), 2.12 (m, 2H) , 2.09 (s, 3H), 1.72 (m, 2H), 1.7/1.19 (m+m, 4H), 1.46 (m, 2H), 0.92 (m, 2H), -0.11 (s, 9H); MSHR-ERI (m/z): [M+2H] ²⁺ , calculated for C ₄₇ H ₆₄ FN ₉ O ₄ S ₂ Si: 465.7211; found: 465.7210.

Step B: 2-[[6-(1,3-benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]-[4-[(1-methyl-4piperidyl)amino]butyl]amino]-5- [3-[4-[3-(dimethylamino)prop-1-ynyl]-2-fluorophenoxy]propyl]thiazole4-carboxylic acid.

Using the general deprotection and hydrolysis procedure followed by repurification by reverse phase preparative chromatography (C18, 0.1% TFA in water:MeCN), the TFA salt of the desired product was obtained from the product from step A.

MSHR-ERI (m/z): [M+2H] ²⁺ , calculated for C ₄₀ H ₄₈ FN ₉ O ₃ S ₂ : 393.6726; found: 393.6735.

Example 182: 2-[[6-(1,3-Benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]-[5-[2(diethylamino)ethylamino]pentyl]amino]-5-[3- [4-[3-(dimethylamino)prop-1-ynyl]-2fluorophenoxy]propyl]thiazole-4-carboxylic acid

N

Step A: methyl 2-[5-[2-(diethylamino)ethylamino]pentyl-[5-methyl-6-[^)-[3-(2trimethylsilylethoxymethyl)-1,3-benzothiazol-2-ylidene]amino]pyridazine -3-yl]amino]-5-[3-[4-[3(dimethylamino)prop-1-ynyl]-2-fluorophenoxy]propyl]thiazole-4-carboxylate.

Using a general alkylation procedure with an in situ formed tosylate, starting from the synthesis product 5f and N,N-diethylethane-1,2-diamine as a suitable amine, the desired product was obtained.

HRMS-ERI (m/z): [M+2H] ²⁺ , calculated for C ₄₈ H ₇₀ FN ₉ O ₄ S ₂ Si: 473.7367; found: 473.7360.

- 225 046336

Step B: 2-[[6-(1,3-benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]-[5-[2(diethylamino)ethylamino]pentyl]amino]-5-[3- [4-[3-(dimethylamino)prop-1-ynyl]-2fluorophenoxy]propyl]thiazole-4-carboxylic acid.

Using the general deprotection and hydrolysis procedure followed by repurification by reverse phase preparative chromatography (C18, 0.1% TFA in water:MeCN), the TFA salt of the desired product was obtained from the product from step A.

MSHR-ERI (m/z): [M+H]+, calculated for C ₄₁ H ₅₃ FN9O ₃ S ₂ : 802.3691; found: 802.3702.

Example 183: 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)-5-(3-{4-[3(dimethylamino )prop-1 -yn-1 -yl]-2-fluorophenoxy}-2-methoxypropyl)-1,3-thiazol-4-carboxylic acid

\

Step A: ethyl 5-(2-methoxy-3-{[tris(propan-2-yl)silyl]oxy}propyl)-2-[methyl(5-methyl-6-{[(2Z)3-{ [ 2-(trimethylsilyl)ethoxy]methyl}-2,3-dihydro-1,3-benzothiazol-2-ylidene]amino}pyridazin-3yl)amino]-1,3-thiazol-4-carboxylate.

Synthesis product 4a (1.6 g, 3.93 mmol, 1 eq.) and synthesis product 3zh (1.75 g, 4.06 mmol, 1.03 eq.) were added to a solution of tris(dibenzylideneacetone)dipalladium(0) (180 mg, 0.2 mmol, 0.05 eq. ) and xantphos (228 mg, 0.39 mmol, 0.1 eq.) in 1,4-dioxane (40 ml). Cesium carbonate (3.84 g, 11.8 mmol, 3 eq.) and N,N-diisopropylethylamine (2.1 mL, 11.8 mmol, 3 eq.) were added and the mixture was heated in a sealed tube at 125°C for 24 h. The suspension was allowed to cool and the mixture was filtered through celite, the solids were washed with 1,4-dioxane (30 ml) and the filtrate was concentrated in vacuo. Purification by flash column chromatography (100 g silica gel) eluting with a gradient of 0-25% ethyl acetate in isoheptane provided the title product as a pale yellow solid (2.49 g, 3.11 mmol, 79%).

LC/MS (C ₃₈ H ₆₀ N ₆ O ₅ Si ₂ S ₂ ) 801 [M+H]+; RT 1.74 (ZHMS-U-V2).

1H NMR (400 MHz, DMSO-dg) δ 7.58 (dt, J=7.6, 0.9 Hz, 1H), 7.42 (d, J=1.2 Hz, 1H), 7.40-7.36 (m, 2H), 7.24-7.17 ( m, 1H), 5.85 (s, 2H), 4.39 (q, J=7.1 Hz, 2H), 3.85 (s, 3H), 3.80 (dd, J=4.8, 2.4 Hz, 2H), 3.78-3.72 (m , 2H), 3.60-3.49 (m, 2H), 3.45 (s, 3H), 3.32-3.15 (m, 1H), 2.46 (d, J=1.0 Hz, 3H), 1.42 (t, J=7.1 Hz, 3H), 1.161.02 (m, 21H), 1.01-0.92 (m, 2H), -0.07 (s, 9H).

Step B: ethyl 5-(3-hydroxy-2-methoxypropyl)-2-[methyl(5-methyl-6-{[(2Z)-3-{[2(trimethylsilyl)ethoxy]methyl}-2,3- dihydro-1,3-benzothiazol-2-ylidene]amino}pyridazin-3-yl)amino]1,3-thiazol-4-carboxylate.

To a solution of the product from step A (2.45 g, 3.06 mmol, 1 eq.) in tetrahydrofuran (15 ml), cooled in ice water, was added tetrabutylammonium fluoride (1 M in tetrahydrofuran; 10 ml, 9.17 mmol, 3 eq.), and the mixture was stirred for 60 min at 0°C. The reaction mixture was partitioned between water (75 ml) and ethyl acetate (150 ml), and the organic phase was washed successively with water (2x75 ml) and brine (75 ml), dried (magnesium sulfate) and concentrated in vacuo. Purification by flash column chromatography (50 g silica gel) eluting with a gradient of 0-50% ethyl acetate in isoheptane gave the title product as a pale yellow/green foam (1.83 g, 2.84 mmol, 93%).

LC/MS (C ₂₉ H ₄₀ N ₆ O ₅ SiS ₂ ) 645 [M+H] ⁺ ; RT 1.56 (LCMS^-B1).

1H NMR (400 MHz, DMSO-dg) δ 7.59 (dt, J=7.6, 0.9 Hz, 1H), 7.40-7.36 (m, 2H), 7.33 (d, J=1.1 Hz, 1H), 7.24-7.18 ( m, 1H), 5.84 (s, 2H), 4.41 (q, J=7.1 Hz, 2H), 3.85 (s, 3H), 3.79-3.70 (m, 2H), 3.47-3.36 (m, 2H), 2.66 (t, J=6.6 Hz, 1H), 2.46 (d, J=1.0 Hz, 3H), 1.43 (t, 3H), 1.03-0.90 (m, 2H), -0.07 (s, 9H).

Step C: ethyl 5-[3-(2-fluoro-4-iodophenoxy)-2-methoxypropyl]-2-[methyl(5-methyl-6-{[(2Z)-3-{[2(trimethylsilyl)ethoxy ] methyl}-2,3-dihydro-1,3-benzothiazol-2-ylidene] amino } pyridazin-3 -yl)amino] 1,3-thiazol-4-carboxylate.

To a solution of the product from step B (750 mg, 1.16 mmol, 1 eq.) in toluene (15 ml) was added diisopropyl azodicarboxylate (0.5 ml, 2.33 mmol, 2 eq.), 2-fluoro-4-iodophenol (500 mg, 2.1 mmol , 1.81 eq.) and triphenylphosphine (610 mg, 2.33 mmol, 2 eq.), and the mixture was heated at 100°C for 18 h, then allowed to cool to ambient temperature and concentrated in vacuo. Purification by flash column chromatography (100 g silica gel) eluting with a gradient of 0-35% ethyl acetate in

- 226 046336 isoheptane gave the target product as an orange resin (1.2 g, 1.04 mmol, 90%).

LC/MS (C ₃₅ H ₄₂ FIN ₆ O ₅ SiS ₂ ) 865 [M+H]+; RT 1.78 (LCMS-V-BI).

¹ H NMR (400 MHz, DMSO) δ 7.60 (d, 1H), 7.42-7.30 (m, 5H), 7.25-7.19 (m, 1H), 6.72 (t, J=8.6 Hz, 1H), 5.85 (s , 2H), 4.40 (q, J=7.1 Hz, 2H), 4.16-4.10 (m, 1H), 4.08-4.01 (m, 1H), 3.98-3.89 (m, 1H), 3.86 (s, 3H), 3.80-3.68 (m, 2H), 3.59-3.49 (m, 4H), 3.48-3.39 (m, 1H), 2.47 (s, 3H), 1.42 (t, J=7.1 Hz, 3H), 1.01-0.92 ( m, 2H), -0.07 (s, 9H).

Step D: ethyl 5-(3-{4-[3-(dimethylamino)prop-1-yn-1-yl]-2-fluorophenoxy}-2-methoxypropyl)-2[methyl(5-methyl-6-{ [(2Z)-3-{[2-(trimethylsilyl)ethoxy]methyl}-2,3-dihydro-1,3-benzothiazol-2ylidene]amino}pyridazin-3-yl)amino]-1,3-thiazol- 4-carboxylate.

To a solution of the product from step C (1.2 g, 1.04 mmol, 1 eq.) in dimethylformamide (15 ml) was added copper(!) iodide (19.8 mg, 0.1 mmol, 0.1 eq.), bis(triphenylphosphine)palladium(II) dichloride (73 mg, 0.1 mmol, 0.1 eq.) and dimethyl(prop-2-in-1-yl)amine (0.25 ml, 2.32 mmol, 2.23 eq.). N,Ndiisopropylethylamine (0.55 mL, 3.12 mmol, 3 eq.) was added and the mixture was heated at 80°C for 90 min. The solution was allowed to cool to ambient temperature and partitioned between ethyl acetate (100 ml) and water (75 ml), and the organic phase was washed successively with water (2x75 ml) and brine (75 ml), dried (magnesium sulfate) and concentrated in vacuo. Purification by flash column chromatography (50 g silica gel) eluting with a gradient of 0-100% ethyl acetate in isoheptane gave the title product as a brown gum (460 mg, 0.56 mmol, 54%).

LC/MS (C ₄₀ H ₅₀ FN ₇ O ₅ SiS ₂ ) 820 [M+H]+; RT 1.52 (LCMS-V-BI).

¹ H NMR (400 MHz, DMSO-de) δ 7.70-7.64 (m, 1H), 7.63-7.58 (m, 1H), 7.42-7.34 (m, 2H), 7.24-7.19 (m, 1H), 7.17- 7.09 (m, 2H), 6.88 (t, J=8.7 Hz, 1H), 5.85 (s, 2H), 4.40 (q, J=7.1 Hz, 2H), 4.19-4.06 (m, 2H), 4.013.89 (m, 1H), 3.86 (s, 3H), 3.80-3.68 (m, 2H), 3.61-3.49 (m, 4H), 3.49-3.37 (m, 3H), 2.47 (t, J=0.9 Hz, 3H ), 2.34 (s, 6H), 1.43 (t, 3H), 1.01-0.92 (m, 2H), -0.07 (d, J=0.7 Hz, 9H).

Step E: ethyl 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)-5-(3-{4[3 -( dimethylamino)prop-1-yn-1-yl]-2-fluorophenoxy}-2-methoxypropyl)-1,3-thiazol-4-carboxylate.

To a solution of the product from step D (460 mg, 0.56 mmol, 1 eq) in dichloromethane (4 ml) was added trifluoroacetic acid (1.1 ml, 14 mmol, 25 eq) and the mixture was stirred for 72 h at ambient temperature. Dichloromethane (150 ml) was added and the mixture was washed successively with 10% aqueous potassium carbonate (75 ml), water (2x75 ml) and brine (75 ml), dried (magnesium sulfate) and concentrated in vacuo. Trituration with diethyl ether (10 ml) resulted in a solid, which was collected by filtration, washed with diethyl ether (2 x 10 ml) and dried in vacuo to give the title product as a green powder (250 mg, 0.33 mmol, 58%).

LC/MS (C ₃₄ H ₃₆ FN ₇ O ₄ S ₂ ) 690 [M+H]+; RT 1.23 (LCMS-V-BI).

1H NMR (400 MHz, DMSO^) δ 7.83-7.69 (m, 2H), 7.62-7.53 (m, 1H), 7.51-7.39 (m, 1H), 7.32-7.25 (m, 1H), 7.25-7.16 ( m, 2H), 6.94 (t, J=8.7 Hz, 1H), 4.47 (q, J=7.1 Hz, 2H), 4.28-4.06 (m, 2H), 4.05-3.95 (m, 1H), 3.95-3.82 (m, 3H), 3.72-3.64 (m, 1H), 3.60 (s, 3H), 3.57-3.52 (m, 1H), 3.50 (s, 2H), 2.51 (s, 3H), 2.41 (s, 6H ), 1.49 (t, J=7.1 Hz, 3H).

Step F: 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)-5-(3-{4-[3(dimethylamino )prop-1-yn-1-yl]-2-fluorophenoxy}-2-methoxypropyl)-1,3-thiazol-4-carboxylic acid.

To a solution of the product from step E (240 mg, 0.31 mmol, 1 eq.) in a 1:2 mixture of 1,4-dioxane (6 ml) was added lithium hydroxide monohydrate (131 mg, 3.13 mmol, 10 eq.) and the suspension was heated at 60°C for 1 hour. The reaction mixture was allowed to cool and filtered through celite and the solids were washed with water (4 ml). The filtrate was acidified with acetic acid to pH 6 to obtain a slurry, and the solids were collected by filtration, washed with water (20 mL) and dried in vacuo to give the title product as a green solid (110 mg, 0.17 mmol, 53%).

MSVR-ERI (m/z) [M+h] ⁺ , calculated for C ₃₂ H ₃₃ FN ₇ O ₄ S ₂ : 662.2014; found: 662.2052.

Example 184: 2-[[6-(1,3-benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]-(4-piperazin-1ylbutyl)amino]-5-[3 -[4-[3 -(dimethylamino)prop-1-ynyl]-2-fluorophenoxy]propyl]thiazole-4-carboxylic acid

Step A: methyl 2-[4-(4-tert-butoxycarbonylpiperazin-1-yl)butyl-[5-methyl-6-[f)-[3-(2

- 227 046336 trimethylsilylethoxymethyl)-1,3-benzothiazol-2-ylidene] amino] pyridazin-3 -yl] amino]-5-[3-[4-[3 (dimethylamino)prop-1-ynyl]-2-fluorophenoxy ]propyl]thiazole-4-carboxylate.

Using a general alkylation procedure with an in situ generated tosylate starting from synthesis 5d and tert-butyl piperazine-1-carboxylate as a suitable amine, the desired product was obtained.

MSHR-ERI (m/z): [M+H]+, calculated for C ₅₀ H ₆₉ FN ₉ O ₆ S ₂ Si: 1002.4560; found: 1002.4544.

Step B: 2-[[6-(1,3-benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]-(4-piperazin-1 ylbutyl)amino]-5-[3-[4-[ 3-(dimethylamino)prop-1-ynyl]-2-fluorophenoxy]propyl]thiazole-4-carboxylic acid.

Using the general deprotection and hydrolysis procedure followed by repurification by reverse phase preparative chromatography (C18, 0.1% TFA in water:MeCN), the TFA salt of the desired product was obtained from the product from step A.

MSHR-ERI (m/z): [M+2H] ²⁺ , calculated for C ₃₈ H ₄₆ FN ₉ O ₃ S ₂ : 379.6569; found: 379.6565.

Example 185: 2-[[6-(1,3-benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]-(4-hydroxybutyl)amino]5-[3-[2-fluoro-4-[ (E)-3-(methylamino)prop-1-enyl]phenoxy]propyl]thiazole-4-carboxylic acid

HN

Step A: tert-butyl N-methyl-N-[(E)-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2yl)allyl]carbamate.

A mixture of 1.50 g tert-butyl N-methyl-N-prop-2-ynylcarbamate (8.86 mmol, 1 eq.), 1.70 g 4,4,5,5tetramethyl-1,3,2-dioxaborolane (13.30 mmol, 1.5 eq. ), 90 mg of N,N-diethylethanamine (0.89 mmol, 0.1 eq.) and 229 mg of chloridobis(n ⁵ -cyclopentadienyl)hydridozirconium (Schwarz reagent) (0.89 mmol, 0.1 eq.) were stirred at 65°C overnight. After completion of the reaction, EtOAc and conc. NH4C1 solution, the organic layer was separated and washed with conc. NaHCO ₃ , conc. NaCl, dried over MgSO ₄ , filtered and then concentrated on celite. The crude product was purified by flash column chromatography using heptane and EtOAc as eluents to obtain 1347 mg (51%) of the title product as a colorless viscous oil.

1H NMR (500 MHz, DMSO-de) δ ppm 6.38 (dt, 1H), 5.35 (dt, 1H), 3.84 (dd, 2H), 2.75 (s, 3H), 1.39 (s, 9H), 1.21 (s, 12H); ¹³ C NMR (125 MHz, DMSOC) δ ppm 155.3, 148.9, 119.1, 83.4, 79.1, 52.2, 34.5, 28.5, 25.1; MSHR-EI (m/z): [M-tBu+H] ⁺ , calculated for CnH ₂₀ BNO ₄ : 241.1480; found: 241.1597.

Step B: methyl 5-[3-[4-[(E)-3-[tert-butoxycarbonyl(methyl)amino]prop-1-enyl]-2fluorophenoxy]propyl]-2-[4-[tert-butyl( dimethyl)silyl]hydroxybutylamino]thiazole-4-carboxylate.

1.00 g of synthesis product 3c, stage B (1.61 mmol, 1 eq.), 716 mg of product from stage A (2.41 mmol, 1.5 eq.), 93 mg Pd(PPh ₃ ) ₄ (0.08 mmol, 0.05 eq.) and 666 mg K2CO3 (4.82 mmol, 3 eq.) was mixed in 15 ml 1,2-dimethoxyethane and 1.9 ml water in a MB flask and irradiated at 110°C for 1 hour. Complete conversion was observed. EtOAc and water were added, the organic layer was separated and washed with conc. NaCl, dried over MgSO ₄ , filtered and then concentrated on celite. The crude product was purified by flash column chromatography using heptane and EtOAc as eluents to obtain 708 mg (66%) of the title product as a colorless viscous oil.

1H NMR (500 MHz, DMSO-dg) δ ppm 7.46 (t, 1H), 7.33 (dd, 1H), 7.14 (dd, 1H), 7.07 (t, 1H), 6.39 (brd, 1H), 6.13 (dt, 1H), 4.06 (t, 2H), 3.90 (brd, 2H), 3.71 (s, 3H), 3.60 (t, 2H), 3.18 (m, 2H), 3.12 (t, 2H), 2.79 (s, 3H), 2.00 (m, 2H), 1.57 ( m, 2H), 1.51 (m, 2H), 1.41 (s, 9H), 0.86 (s, 9H), 0.02 (s, 6H); ¹³ C NMR (125 MHz, DMSO-ds) δ ppm 164.8, 163.0, 155.3, 152.5, 146.3, 137.0, 136.1, 130.7, 130.5, 125.3, 123.4, 115.7, 113.8, 79.1, 68.4, 62.7, 51.6, 50.5, 44 .5, 34.1, 30.9, 30.2, 28.6, 26.3, 25.7, 23.4, 18.4, -4.8; MSHR-ERI (m/z): [M+H] ⁺ , calculated for C ₃₃ H ₅₃ FN ₃ O ₆ SSi: 666.3403; found: 666.3409.

Step C: methyl 5-[3-[4-[(E)-3-[tert-butoxycarbonyl(methyl)amino]prop-1-enyl]-2fluorophenoxy]propyl]-2-[4-[tert-butyl( dimethyl)silyl]oxybutyl-[5-methyl-6-[^)-[3-(2trimethylsilylethoxymethyl)-1,3-benzothiazol-2-ylidene]amino]pyridazin-3-yl]amino]thiazol-4carboxylate.

Using the general Buchwald reaction procedure II, starting from 138 mg of the product from step B (0.21 mmol, 1 eq.) and 101 mg of the product of synthesis 4a (0.25 mmol, 1.2 eq.) as the appropriate halide, 102 mg (48%) were obtained target product.

- 228 046336

1H NMR (500 MHz, DMSO-dg) δ ppm 7.81 (dm, 1H), 7.62 (brs, 1H), 7.46 (dm, 1H), 7.43 (m, 1H), 7.33 (ddd, 1H), 7.25 (m, 1H), 7.15 (dm, 1H), 7.10 (t, 1H), 6.38 (d, 1H), 6.10 (dt, 1H), 5.86 (s, 2H), 4.42 (t, 2H), 4.14 (t, 2H), 3.88 (d, 2H), 3.79 ( s, 3H), 3.75 (m, 2H), 3.63 (t, 2H), 3.27 (t, 2H), 2.77 (s, 3H), 2.45 (d, 3H), 2.12 (m, 2H), 1.77 (m , 2H), 1.54 (m, 2H), 1.40 (s, 9H), 0.92 (m, 2H), 0.81 (s, 3H), -0.01 (s, 6H), -0.09 (s, 9H); MSVRERI (m/z): [M+H]+, calculated for C ₅₁ H ₇₅ FN ₇ O ₇ S ₂ Si ₂ : 1036.4686; found: 1036.4706.

Step D: 2-[[6-(1,3-benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]-(4-hydroxybutyl)amino]-5[3 - [2-fluoro-4-[ (E)-3-(methylamino)prop-1-enyl]phenoxy]propyl]thiazole-4-carboxylic acid.

Using the general deprotection and hydrolysis procedure followed by repurification by reverse phase preparative chromatography (C18, 0.1% TFA in water:MeCN), the product from step C gave the TFA salt of the desired product with an isomer ratio of ca. 82% E and 18% Z (based on NMR data).

MSHR-ERI (m/z): [M+H]+, calculated for C ₃₃ H ₃₇ FN ₇ O ₄ S ₂ : 678.2327; found: 678.2326.

Example 186: 2-[[6-(1,3-benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]-(4-hydroxybutyl)amino]5-[3-[2-fluoro-4-[ 3-(methylamino)propyl]phenoxy]propyl]thiazole-4-carboxylic acid

HN

Step A: methyl 5-[3-[4-[3-[tert-butoxycarbonyl(methyl)amino]propyl]-2-fluorophenoxy]propyl]-2[4-[tert-butyl(dimethyl)silyl]oxybutylamino]thiazole -4-carboxylate.

A 20 mL oven-dried pressure reaction vessel equipped with a PTFE-coated magnetic stir bar was charged with 400 mg of Example 185, Step B (0.60 mmol, 1 eq.), 64 mg of 10 wt.% Pd/C ( 0.06 mmol, 0.1 eq.) in 6 ml of methanol, and then it was created under a nitrogen atmosphere using a hydrogenation system. The vessel was then filled with H ₂ gas at a pressure of 4 bar and the reaction mixture was stirred at room temperature. for 3 hours. Complete conversion was observed, celite was added to the reaction mixture, and the volatiles were removed under reduced pressure. The crude product was then purified by flash column chromatography using heptane and EtOAc as eluents to obtain 381 mg (95%) of the title product as a colorless oil.

1H NMR (500 MHz, DMSO-dg) δ ppm 7.57 (t, 1H), 7.07 (dd, 1H), 7.02 (t, 1H), 6.93 (d, 1H), 4.01 (t, 2H), 3.69 (s, 3H), 3.59 (t, 2H), 3.17 (q, 2H), 3.13 (m, 2H), 3.11 (t, 2H), 2.75 (brs, 3H), 2.47 (t, 2H), 1.98 (quin, 2H), 1.72 (br, 2H), 1.54 ( m, 2H), 1.50 (m, 2H), 1.38/1.33 (brs+brs, 9H), 0.85 (s, 9H), 0.01 (s, 6H); ¹³ C NMR (125 MHz, DMSO-dg) δ ppm 164.6, 163.1, 155.2, 152.0, 144.7, 136.5, 135.3, 124.6, 116.2, 115.3, 79.0, 68.1, 62.7, 51.7, 47.9, 44.4, 34.1, 31.7, 30.9, 30.1, 29.3, 28.6, 26.3, 25.7, 23.4, 18.5, -4.9; MSHR-ERI (m/z): [M+H]+, calculated for C ₃₃ H55FN ₃ O(5SSi: 668.3559; found: 668.3544.

Step B: methyl 5-[3-[4-[3-[tert-butoxycarbonyl(methyl)amino]propyl]-2-fluorophenoxy]propyl]-2[4-[tert-butyl(dimethyl)silyl]oxybutyl-[ 5-methyl-6-[^)-[3-(2-trimethylsilylethoxymethyl)-1,3benzothiazol-2-ylidene]amino]pyridazin-3-yl]amino]thiazol-4-carboxylate.

Using the general Buchwald reaction procedure II, starting with 375 mg of the product from step A (0.56 mmol, 1 eq.) and 274 mg of the product of synthesis 4a (0.67 mmol, 1.2 eq.) as the appropriate halide, 479 mg (82%) were obtained target product.

1H NMR (500 MHz, DMSO^) δ ppm 7.80 (dm, 1H), 7.65/7.61 (s/s, 1H), 7.46 (dd, 1H), 7.43 (td, 1H), 7.24 (td, 1H), 7.05 (t, 1H), 7.04 (m, 1H), 6.93 (dm, 1H), 5.85 (s, 2H), 4.41 (t, 2H), 4.10 (t, 2H), 3.80/3.79 (s/s, 3H), 3.74 (m, 2H), 3.63 /3.47 (t, 2H), 3.26 (m, 2H), 3.13 (t, 2H), 2.74 (s, 3H), 2.47 (t, 2H), 2.46/2.45 (s/s, 3H), 2.11 (m , 2H), 1.77 (m, 2H), 1.72 (m, 2H), 1.54 (m, 2H), 1.35 (br., 9H), 0.92 (m, 2H), 0.85/0.81 (s/s, 9H) , -0.01/-0.03 (s, 6H), -0.09/-0.1 (s, 9H); ¹³ C NMR (125 MHz, DMSO^) δ ppm 155.4/155.3, 127.1, 124.6, 123.4, 123.1, 117.8, 116.2, 116.1, 111.9, 78.7, 73.1, 68.9, 66.8, 62.6/61, 51.8, 48.0, 46.9, 34. 2, 31.8, 31.2, 29.8, 29.2, 28.5, 26.3/26.2, 23.9, 23.2, 17.9, 17.7, -1.0, -2.7/-4.9; HRMS-ERI (m/z): [M+H]+, calculated for C ₅₁ H ₇₇ FN ₇ O ₇ S ₂ Si ₂ : 1038.4843; found: 1038.4837.

Step C: 2-[[6-(1,3-benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]-(4-hydroxybutyl)amino]-5[3-[2-fluoro-4-[ 3-(methylamino)propyl]phenoxy]propyl]thiazole-4-carboxylic acid.

Using the general deprotection and hydrolysis procedure followed by repurification by reverse phase preparative chromatography (C18, 0.1% TFA in water:MeSU), the TFA salt of the desired product was obtained from the product from step B.

MSHR-ERI (m/z): [M+H]+, calculated for C ₃₃ H ₃₉ FN ₇ O ₄ S ₂ : 680.2483; found: 680.2469.

- 229 046336

Example 187: 2-[[6-(1,3-Benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]-[5-[2-(1piperidyl)ethylamino]pentyl]amino]-5-[3 -[4-[3-(dimethylamino)prop-1-ynyl]-2fluorophenoxy]propyl]thiazole-4-carboxylic acid

Step A: methyl 5-[3-[4-[3-(dimethylamino)prop-1-ynyl]-2-fluorophenoxy]propyl]-2-[[5-methyl-6[^)-[3-(2 -trimethylsilylethoxymethyl)-1,3-benzothiazol-2-ylidene]amino]pyridazin-3-yl]-[5-[2-(1piperidyl)ethylamino]pentyl]amino]thiazole-4-carboxylate.

Using a general alkylation procedure with an in situ formed tosylate starting from the synthesis product 5f and 2-(1-piperidyl)ethanamine as a suitable amine, the desired product was obtained.

HRMS-ERI (m/z): [M+2H]2+, calculated for C ₄₉ H ₇₀ FN ₉ O ₄ S ₂ Si: 479.7367; found: 479.7353.

Step B: 2-[[6-(1,3-benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]-[5-[2-(1piperidyl)ethylamino]pentyl]amino]-5-[3 -[4-[3-(dimethylamino)prop-1-ynyl]-2fluorophenoxy]propyl]thiazole-4-carboxylic acid.

Using the general deprotection and hydrolysis procedure followed by repurification by reverse phase preparative chromatography (C18, 0.1% TFA in water:MeCN), the TFA salt of the desired product was obtained from the product from step A.

MSHR-ERI (m/z): [M+H]+, calculated for C ₄₂ H ₅₃ FN ₉ O ₃ S ₂ : 814.3691; found: 814.3670.

Example 188: 2-[[6-(1,3-benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]-[4(methylamino)butyl]amino]-5-[3-[4-[3 -(dimethylamino)prop-1-ynyl]-2-fluorophenoxy]propyl]thiazole-4carboxylic acid

Step A: methyl 5-[3-[4-[3-(dimethylamino)prop-1-ynyl]-2-fluorophenoxy]propyl]-2-[4(methylamino)butyl-[5-methyl-6-[^ )-[3-(2-trimethylsilylethoxymethyl)-1,3-benzothiazol-2ylidene]amino]pyridazin-3-yl]amino]thiazol-4-carboxylate.

Using a general alkylation procedure with an in situ tosylate starting from synthesis 5d and methanamine as a suitable amine, the desired product was obtained.

1H NMR (500 MHz, DMSO-06) δ ppm 7.84 (dm, 1H), 7.72 (s, 1H), 7.47 (dm, 1H), 7.44 (m, 1H), 7.31 (dd, 1H), 7.25 (m, 1H), 7.21 (dm, 1H), 7.16 (t, 1H), 5.86 (s, 2H), 4.37 (t, 2H), 4.15 (t, 2H), 3.78 (s, 3H), 3.72 (m, 2H), 3.38 (s, 2H), 3.27 ( t, 2H), 2.52 (t, 2H), 2.46 (s, 3H), 2.26 (s, 3H), 2.19 (s, 6H), 2.13 (m, 2H), 1.71 (m, 2H), 1.49 (m , 2H), 0.92 (m, 2H), -0.11 (s, 9H); MSHR-ERI (m/z): [M+H]+, calculated for C^^FNsO^Si: 847.3614; found: 847.3594.

Step B: 2-[[6-(1,3-benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]-[4(methylamino)butyl]amino]-5-[3-[4-[3 -(dimethylamino)prop-1-ynyl]-2-fluorophenoxy]propyl]thiazole-4carboxylic acid.

Using the general deprotection and hydrolysis procedure followed by repurification by reverse phase preparative chromatography (C18, 0.1% TFA in water:MeCN), the TFA salt of the desired product was obtained from the product from step A.

MSHR-ERI (m/z): [M+2H] ²⁺ , calculated for C ₃₅ H ₄₁ FN ₈ O ₃ S ₂ : 352.1358; found: 352.1361.

Example 189: 2-[[6-(1,3-benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]-(5-piperazin-1

- 230 046336 ylpentyl)amino]-5-[3-[4-[3-(dimethylamino)prop-1-ynyl]-2-fluorophenoxy]propyl]thiazole-4-carboxylic acid

Step A: methyl 2-[5-(4-tert-butoxycarbonylpiperazin-1-yl)pentyl-[5-methyl-6-[^)-[3-(2trimethylsilylethoxymethyl)-1,3-benzothiazol-2-ylidene] amino]pyridazin-3-yl]amino]-5-[3-[4-[3(dimethylamino)prop-1-ynyl]-2-fluorophenoxy]propyl]thiazole-4-carboxylate.

Using a general alkylation procedure with an in situ formed tosylate, starting from the synthesis product 5f and tert-butyl piperazine-1-carboxylate as a suitable amine, the desired product was obtained.

MSHR-ERI (m/z): [M+H]+, calculated for C ₅₁ H ₇₁ FN ₉ O ₆ S ₂ Si: 1016.4716; found: 1016.4710.

Step B: 2-[[6-(1,3-benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]-(5-piperazin-1ylpentyl)amino]-5-[3-[4-[3 -(dimethylamino)prop-1-ynyl]-2-fluorophenoxy]propyl]thiazole-4-carboxylic acid.

Using the general deprotection and hydrolysis procedure followed by repurification by reverse phase preparative chromatography (C18, 0.1% TFA in water:MeCN), the TFA salt of the desired product was obtained from the product from step A.

MSHR-ERI (m/z): [M+2H]2 ⁺ , calculated for C ₃₉ H ₄₈ FN ₉ O ₃ S ₂ : 386.6647; found: 386.6642.

Example 190: 2-[[6-(1,3-Benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]-[5-[methyl-[2(methylamino)ethyl]amino]pentyl]amino]- 5-[3-[4-[3-(dimethylamino)prop-1-ynyl]-2fluorophenoxy]propyl]thiazole-4-carboxylic acid

Step A: methyl 2-[5-[2-[tert-butoxycarbonyl(methyl)amino]ethylmethylamino]pentyl-[5-methyl-6[^)-[3-(2-trimethylsilylethoxymethyl)-1,3-benzothiazol- 2-ylidene]amino]pyridazin-3-yl]amino]-5-[3-[4[3-(dimethylamino)prop-1-ynyl]-2-fluorophenoxy]propyl]thiazole-4-carboxylate.

Using the general alkylation procedure with an in situ formed tosylate, starting from the synthesis product 5f and tert-butyl Nmethyl-Na[2-(methylamino)ethyl]carbamate as a suitable amine, the desired product was obtained.

¹ H NMR (500 MHz, DMSO-de) δ ppm 7.84 (d, 1H), 7.66 (s, 1H), 7.47 (d, 1H), 7.43 (td, 1H), 7.3 (dd, 1H), 7.25 (td, 1H), 7.20 (dd, 1H), 7.15 (t, 1H), 5.86 (s, 2H), 4.37 (t, 2H), 4.14 (t, 2H), 3.77 (s, 3H), 3.72 (m, 2H), 3.38 (s, 2H), 3.26 ( t, 2H), 3.16 (brt., 2H), 2.73 (s, 3H), 2.45 (s, 3H), 2.35 (brt., 2H), 2.28 (brt., 2H), 2.19 (s, 6H), 2.13 (s, 3H), 2.12 (m, 2H), 1.68 (m, 2H), 1.45 (m, 2H), 1.35 (m, 2H), 1.33 (s, 9H), 0.92 (m, 2H), - 0.11 (s, 9H); ¹³ C NMR (125 MHz, DMSO-d^ δ ppm 129.0, 127.2, 123.4, 123.2, 119.2, 117.6, 115.5, 112.0, 72.9, 68.4, 66.7, 57.4, 55.2, 51.9, 48 .1, 46.8, 46.4, 44.2, 42.6, 34.5, 31.0, 28.5, 27.0, 26.8, 24.1, 23.1, 17.8, 17.8, -1.0; HRMS-ERI (m/z): [M+H] ⁺ , calculated for C ₅₁ H ₇₃ FN ₉ O ₆ S ₂ Si: 1018.4873; found: 1018.4869.

Step B: 2-[[6-(1,3-benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]-[5-[methyl-[2(methylamino)ethyl]amino]pentyl]amino]- 5-[3-[4-[3-(dimethylamino)prop-1-ynyl]-2fluorophenoxy]propyl]thiazole-4-carboxylic acid.

- 231 046336

Using the general deprotection and hydrolysis procedure followed by repurification by reverse phase preparative chromatography (C18, 0.1% TFA in water:MeCN), the TFA salt of the desired product was obtained from the product from step A.

MSHR-ERI (m/z): [M+2H] ²⁺ , calculated for C ₃₉ H ₅₀ FN9O ₃ S ₂ : 387.6726; found: 387.6731.

Example 191: 2-[5-Aminopentyl-[6-(1,3-benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]amino]-5[3-[4-[3-(dimethylamino)prop -1-ynyl]-2-fluorophenoxy]propyl]thiazole-4-carboxylic acid

Step A: methyl 2-[tert-butoxycarbonyl-[5-(tert-butoxycarbonylamino)pentyl]amino]-5-[3-[4[3-(dimethylamino)prop-1-ynyl]-2-fluorophenoxy]propyl] thiazole-4-carboxylate.

Using the general Mitsunobu reaction procedure, starting from 250 mg of synthesis product 1c (0.51 mmol, 1 eq.) and 207 mg of tert-butyl H-(5-hydroxypentyl)carbamate (1.02 mmol, 2 eq.) as a suitable alcohol, we obtained 189 mg (55%) of the target product.

1H NMR (400 MHz, DMSO-dg) δ ppm 7.29 (dd, 1H), 7.21 (d, 1H), 7.13 (t, 1H), 6.75 (t, 1H), 4.10 (t, 2H), 3.97 (t, 2H), 3.75 (s, 3H), 3.49 (br.s, 2H), 3.22 (t, 2H), 2.90 (q, 2H), 2.08 (qv, 2H), 1.99 (s, 6H), 1.66-1.56 (m, 2H), 1.50 (s, 9H ), 1.44-1.36 (m, 2H), 1.35 (s, 9H), 1.28-1.20 (m, 2H); LC-MS-ESI (m/z): [M+H] ⁺ , calculated for C ₃ 4H ₅ 0FN4O ₇ S: 677.3; found: 677.4.

Step B: Methyl 2-[5-(tert-butoxycarbonylamino)pentylamino]-5-[3-[4-[3-(dimethylamino)prop-1ynyl]-2-fluorophenoxy]propyl]thiazole-4-carboxylate.

Using the general HFIP deprotection procedure, starting from 182 mg of the product from Step A (0.27 mmol, 1 eq.) as the appropriate Boc-protected amine, 106 mg (68%) of the title product was obtained.

1H NMR (400 MHz, DMSO^) δ ppm 7.56 (t, 1H), 7.30 (dd, 1H), 7.21 (d, 1H), 7.12 (t, 1H), 6.77 (t, 1H), 4.07 (t, 2H), 3.69 (s, 3H), 3.44 (s, 2H), 3.16-3.09 (m, 4H), 2.92-2.86 (m, 2H), 2.25 (s, 6H), 2.04-1.97 (m, 2H), 1.53-1.45 (m, 2H), 1.40 -1.33 (m, 2H), 1.36 (s, 9H), 1.31-1.24 (m, 2H); LC-MS-ESI (m/z): [M+H] ⁺ , calculated for C ₂ 9H4 ₂ FN4O ₅ S: 577.3; found: 577.4.

Step C: methyl 2-[5-(tert-butoxycarbonylamino)pentyl-[5-methyl-6-[^)-[3-(2trimethylsilylethoxymethyl)-1,3-benzothiazol-2-ylidene]amino]pyridazin-3- yl]amino]-5-[3-[4-[3(dimethylamino)prop-1-ynyl]-2-fluorophenoxy]propyl]thiazole-4-carboxylate.

Using the general Buchwald reaction procedure II, starting from 102 mg of the product from step B (0.18 mmol, 1 eq.) and 90 mg of the product of synthesis 4a (0.22 mmol, 1.25 eq.) as the appropriate halide, 102 mg (61%) were obtained target product.

1H NMR (500 MHz, DMSO^) δ ppm 7.84 (d, 1H), 7.67 (s, 1H), 7.47 (d, 1H), 7.44 (td, 1H), 7.31 (dd, 1H), 7.25 (td, 1H), 7.21 (dm, 1H), 7.16 (t, 1H), 6.77 (t, 1H), 5.86 (s, 2H), 4.34 (t, 2H), 4.15 (t, 2H), 3.78 (s, 3H), 3.72 (t, 2H), 3.39 ( s, 2H), 3.27 (t, 2H), 2.92 (q, 2H), 2.47 (s, 3H), 2.21 (s, 6H), 2.13 (m, 2H), 1.69 (m, 2H), 1.44 (m , 2H), 1.35 (s, 18H), 1.34 (m, 2H), 0.92 (t, 2H), -0.11 (s, 9H); MSHR-ERI (m/z): [M+H] ⁺ , calculated for C4 ₇ H64FN8O6S ₂ Si: 947.4138; found: 947.4133.

Step D: 2-[5-aminopentyl-[6-(1,3-benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]amino]-5-[3[4-[3-(dimethylamino)prop -1-ynyl]-2-fluorophenoxy]propyl]thiazole-4-carboxylic acid.

Using the general deprotection and hydrolysis procedure, the desired product was obtained from the product from step C.

MSHR-ERI (m/z): [M+2H] ²⁺ , calculated for C ₃₅ H ₄₁ FN ₈ O ₃ S ₂ : 352.1364; found: 352.1370.

Example 192: 2-[4-Aminobutyl-[6-(1,3-benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]amino]-5-[3[4-[3-(dimethylamino)prop -1-ynyl]-2-fluorophenoxy]propyl]thiazole-4-carboxylic acid

- 232 046336

Step A: methyl 2-[tert-butoxycarbonyl-[4-(tert-butoxycarbonylamino)butyl]amino]-5-[3-[4-[3(dimethylamino)prop-1-ynyl]-2-fluorophenoxy]propyl] thiazole-4-carboxylate.

Using the general Mitsunobu reaction procedure, starting from 250 mg of synthesis product 1c (0.51 mmol, 1 eq.) and 193 mg of tert-butyl X-(4-hydroxybutyl)carbamate (1.02 mmol, 2 eq.) as a suitable alcohol, we obtained 220 mg (65%) of the target product.

1H NMR (400 MHz, DMSOD6) δ ppm 7.30 (dd, 1H), 7.21 (d, 1H), 7.13 (t, 1H), 6.80 (t, 1H), 4.10 (t, 2H), 4.01-3.95 (m, 2H), 3.75 (s, 3H) , 3.45 (s, 2H), 3.22 (t, 2H), 2.91 (q, 2H), 2.25 (s, 6H), 2.08 (qv, 2H), 1.63-1.54 (m, 2H), 1.50 (s, 9H ), 1.40-1.35 (m, 2H), 1.35 (s, 9H); LC-MS-ESI (m/z): [M+H]+, calculated for C ₃₃ H ₄₈ FN ₄ O ₇ S: 663.3; found: 663.4.

Step B: Methyl 2-[4-(tert-butoxycarbonylamino)butylamino]-5-[3-[4-[3-(dimethylamino)prop-1ynyl]-2-fluorophenoxy]propyl]thiazole-4-carboxylate.

Using the general HFIP deprotection procedure, starting with 215 mg of the product from Step A (0.33 mmol, 1 eq.) as the appropriate Boc-protected amine, 137 mg (75%) of the desired product was obtained.

1H NMR (400 MHz, DMSOD6) δ ppm 7.57 (t, 1H), 7.30 (d, 1H), 7.21 (d, 1H), 7.12 (t, 1H), 6.81 (t, 1H), 4.07 (t, 2H), 3.69 (s, 3H), 3.42 (s, 2H), 3.17-3.09 (m, 4H), 2.94-2.88 (m, 2H), 2.23 (s, 6H), 2.04-2.00 (m, 2h), 1.53-1.37 (m, 4H), 1.36 (s, 9H); LC-MS-ESI (m/z): [M+H]+, calculated for C ₂₈ H ₄₀ FN ₄ O ₅ S: 563.3; found: 563.2.

Step C: methyl 2-[4-(tert-butoxycarbonylamino)butyl-[5-methyl-6-[^)-[3-(2trimethylsilylethoxymethyl)-1,3-benzothiazol-2-ylidene]amino]pyridazin-3- yl]amino]-5-[3-[4-[3(dimethylamino)prop-1-ynyl]-2-fluorophenoxy]propyl]thiazole-4-carboxylate.

Using the general Buchwald reaction procedure II, starting from 133 mg of the product from step B (0.24 mmol, 1 eq.) and 120 mg of the product of synthesis 4a (0.29 mmol, 1.25 eq.) as the appropriate halide, 220 mg (98%) were obtained target product.

1H NMR (500 MHz, DMSO-dg) δ ppm 7.84 (d, 1H), 7.69 (s, 1H), 7.47 (d, 1H), 7.44 (td, 1H), 7.31 (dd, 1H), 7.25 (td, 1H), 7.21 (dm, 1H), 7.16 (t, 1H), 6.82 (t, 1H), 5.86 (s, 2H), 4.36 (t, 2H), 4.15 (t, 2H), 3.78 (s, 3H), 3.72 (t, 2H), 3.38 ( s, 2H), 3.27 (t, 2H), 2.98 (q, 2H), 2.46 (s, 3H), 2.19 (s, 6H), 2.13 (m, 2H), 1.67 (m, 2H), 1.46 (m , 2H), 1.34 (s, 9H), 0.92 (t, 2H), -0.11 (s, 9H); ¹³ C NMR (125 MHz, DMSO-dg) δ ppm 156.1, 128.9, 127.2, 123.5, 123.2, 119.2, 117.6, 115.5, 112.0, 72.9, 68.4, 66.7, 52.0, 48.1, 46.7, 44.2, 39.8, 31.0, 28.7, 27.2, 24.7, 23.1, 17.9, 17.8, -1.0 ; MSHR-ERI (m/z): [M+H]+, calculated for C46H62FN8O6S2SE 933.3982; found: 933.3995.

Step D: 2-[4-aminobutyl-[6-(1,3-benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]amino]-5-[3-[4[3-(dimethylamino)prop -1-ynyl]-2-fluorophenoxy]propyl]thiazole-4-carboxylic acid.

Using the general deprotection and hydrolysis procedure followed by repurification by reverse phase preparative chromatography (C18, 0.1% TFA in water-MeSA), the TFA salt of the desired product was obtained from the product from step C.

MSHR-ERI (m/z): [M+2H] ²⁺ , calculated for C ₃₄ H ₃₉ FN ₈ O ₃ S ₂ : 345.1280; found: 345.1281.

Example 193: 2-[[6-(1,3-benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]-[5-(dimethylamino)-4hydroxypentyl]amino]-5-[3-[2- fluoro-4-[3-(methylamino)prop-1-ynyl]phenoxy]propyl]thiazole-4carboxylic acid

- 233 046336

Using the general technique for deprotecting a quaternary salt, starting from the example compound

144, stage A, the target product was obtained.

MSHR-ERI (m/z): [M+H]+, calculated for C ₃₆ H ₄₂ FN ₈ O ₄ S2: 733.2749; found: 733.2745.

Example 194: 2-[[6-(1,3-Benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]-[5(methylamino)pentyl] amino]-5-[3-[4-[3 -(dimethylamino)prop-1-ynyl]-2-fluorophenoxy]propyl]thiazole-4carboxylic acid

Step A: methyl 5-[3-[4-[3-(dimethylamino)prop-1-ynyl]-2-fluorophenoxy]propyl]-2-[5(methylamino)pentyl-[5-methyl-6-[^ )-[3-(2-trimethylsilylethoxymethyl)-1,3-benzothiazol-2ylidene]amino]pyridazin-3-yl]amino]thiazol-4-carboxylate.

Using a general alkylation procedure with an in situ tosylate starting from synthesis 5f and methamine as a suitable amine, the desired product was obtained after concentration and carried directly to the next step without any further purification.

Step B: 2-[[6-(1,3-benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]-[5(methylamino)pentyl] amino]-5-[3-[4-[3 -(dimethylamino)prop-1-ynyl]-2-fluorophenoxy]propyl]thiazole-4carboxylic acid.

Using the general deprotection and hydrolysis procedure followed by repurification by reverse phase preparative chromatography (C18, 0.1% TFA in water:MeCN), the TFA salt of the desired product was obtained from the crude product from Step A.

MSVR-ERI (m/z): [M+H] ⁺ , calculated for C36H42FN8O3S2: 717.2800; found: 717.2785.

Example 195: 2-[[6-(1,3-Benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]-[5-[3-(4-methylpiperazin-1yl)propylamino]pentyl] amino] - 5 -[3-[4-[3-(dimethylamino)prop-1-ynyl]-2-fluorophenoxy]propyl]thiazole4-carboxylic acid

Step A: methyl 5-[3-[4-[3-(dimethylamino)prop-1-ynyl]-2-fluorophenoxy]propyl]-2-[5-[3-(4methylpiperazin-1-yl)propylamino]pentyl -[5-methyl-6-[^)-[3-(2-trimethylsilylethoxymethyl)-1,3benzothiazol-2-ylidene]amino]pyridazin-3-yl]amino]thiazol-4-carboxylate.

Using a general alkylation procedure with an in situ formed tosylate, starting from the product syn

- 234 046336 thesis 5f and 3-(4-methylpiperazin-1-yl)propan-1-amine as a suitable amine, the desired product was obtained.

1H NMR (400 MHz, DMSO-0 ₆ ) δ ppm 7.83 (dm, 1H), 7.65 (s, 1H), 7.42-7.46 (m, 2H), 7.36-7.08 (m, 4H), 5.85 (s, 2H), 4.36 (t, 2H), 4.14 (t, 2H), 3.78 (s, 3H), 3.72 (t, 2H), 3.26 (t, 2H), 2.70-2.00 (m, 26H), 2.45 (s, 3H), 1.80-1.30 (m, 8H), 0.91 (t, 2H), -0.12 (s, 9H); LC-MS-ESI (m/z): [M+H]+, calculated for C50H72FN10O ₄ S2Si: 987.5; found: 987.4.

Step B: 2-[[6-(1,3-benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]-[5-[3-(4-methylpiperazin-1yl)propylamino]pentyl]amino]- 5-[3-[4-[3-(dimethylamino)prop-1-ynyl]-2-fluorophenoxy]propyl]thiazole4-carboxylic acid.

Using the general deprotection and hydrolysis procedure followed by repurification by reverse phase preparative chromatography (C18, 0.1% TFA in water:MeCN), the TFA salt of the desired product was obtained from the product from step A.

MSHR-ERI (m/z): [M+H]+, calculated for C43H56FN10O3S2: 843.3962; found: 843.3943.

Example 196: 2-[[6-(1,3-Benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]-[4-hydroxy-5-[methyl(2phosphonoethyl)amino]pentyl]amino]-5 -[3-[2-fluoro-4-[3-(methylamino)prop-1-ynyl]phenoxy]propyl]thiazole4-carboxylic acid

Stage A: 2-diethoxyphosphoryl-N-methylethanamine

3.28 g of 1-[ethoxy(vinyl)phosphoryl]oxyethane (20 mmol, 1.0 eq.) and 25 ml of methamine (33 wt.% in EtOH, 200 mmol, 100 eq.) were mixed in a sealed tube and stirred at room temperature. within 20 hours. Complete transformation was observed. The reaction mixture was concentrated to give 3.87 g (99%) of crude title product.

¹ H NMR (400 MHz, DMSOHO δ ppm 4.04-3.90 (m, 4H), 2.67-2.58 (m, 2H), 2.25 (s, 3H), 2.18-1.95 (brs, 1H), 1.93-1.82 (m, 2H), 1.22 (t, 6H); ¹³ C NMR (100 MHz, DMSO-0 ₆ ) δ ppm 60.9, 60.8, 45.03, 45.01, 35.6, 26.1, 24.7, 16.32, 16.26; GC- TOF (EI) (m/z): [M] ⁺ , calculated for C7H18NO3P: 195.1024; found: 195.1009.

Step B: methyl 5-[3-[4-[3-[tert-butoxycarbonyl(methyl)amino]prop-1-ynyl]-2fluorophenoxy]propyl]-2-[[4-[tert-butyl(diphenyl)silyl ]oxy-5-[2diethoxyphosphorylethyl(methyl)amino]pentyl]-[5-methyl-6-[^)-[3-(2-trimethylsilylethoxymethyl)-1,3benzothiazol-2-ylidene]amino]pyridazin-3-yl ]amino]thiazole-4-carboxylate.

Using the general tosylate alkylation procedure, starting from the synthesis product 5a and the product from step A as a suitable amine, the desired product was obtained.

MSHR-ERI (m/z): [M+H]+, calculated for C ₆₉ H ₉₅ FN ₈ O ₁₀ PS ₂ Si ₂ : 1365.5867; found: 1365.5916.

Step C: 2-[[6-(1,3-benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]-[4-hydroxy-5-[methyl(2phosphonoethyl)amino]pentyl]amino]-5 -[3-[2-fluoro-4-[3-(methylamino)prop-1-ynyl]phenoxy]propyl]thiazole4-carboxylic acid.

Using the general procedure for deprotection and hydrolysis of phosphonic acid derivatives, starting from the product from step B, the desired product was obtained.

MSVR-ERI (m/z): [M+H]+, calculated for C3 ₇ H4 ₅ FN ₈ O ₇ PS ₂ : 827.2569; found: 827.2561.

Example 197: 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)-5-[3-({4fluoro-6-[ 3 -(methylamino)prop-1 -yn-1 -yl]pyridin-3 -yl}oxy)propyl]-1,3-thiazol-4-carboxylic acid

ABOUT

- 235 046336

Step A: 2-chloro-4-fluoro-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridine.

To a solution of 2-chloro-4-fluoropyridine (2.5 g; 19.01 mmol) in tetrahydrofuran (30 ml), bis(pinacolato)diboron (2.41 g; 9.50 mmol; 0.5 eq.), 4,4'-di-tert- butyl-2,2'-dipyridyl (51.0 mg; 0.190 mmol; 0.01 eq.) and (1,5-cyclooctadiene)(methoxy)iridium(1) dimer (63.0 mg; 0.095 mmol; 0.005 eq.). The reaction mixture was purged with argon and stirred at 80°C for 16 hours, and then concentrated to dryness. The resulting residue was purified by silica gel column chromatography using cyclohexane/ethyl acetate as eluent to give the title compound (3.64 g; 74%).

1H NMR (DMSO δ ppm 1.33 (s, 12H), 7.58 (d, 1H, J=8.8 Hz), 8.55 (d, 1H, J=9.2 Hz).

Step B: 6-chloro-4-fluoropyrcdin-3-ol.

To a solution of the product from step A (2.00 g; 7.77 mmol) in tetrahydrofuran (16 ml) at 0°C, sodium hydroxide (311 mg; 7.77 mmol; 1.0 eq.) and a 33% aqueous solution of hydrogen peroxide (2.38 ml; 23.3 eq.) were successively added mmol; 3.0 eq.). The reaction mixture was stirred at 0°C for 3 hours and at room temperature. for 16 hours. The reaction mixture was diluted with water (50 ml) and acidified with 1 N. aqueous solution of hydrochloric acid to pH=1. The aqueous layer was extracted with dichloromethane (3x100 ml) and the combined organic layers were dried over sodium sulfate, filtered and concentrated to dryness. The resulting residue was purified by silica gel column chromatography using cyclohexane/ethyl acetate as eluent to give the title compound (640 mg; 56%).

1H NMR (DMSO δ ppm 7.53 (d, 1H, J=10.4 Hz), 8.06 (d, 1H, J=10.8 Hz), 10.76 (s, 1H).

Step C: tert-butyl N[3-(4-fluoro-5-hydroxy-2-pyridyl)prop-2-ynyl]-N-methylcarbamate.

To a solution of the product from stage B (300 mg; 2.03 mmol) in acetonitrile (12 ml), tert-butyl N-methyl-N-prop-2-ynylcarbamate (688 mg; 4.07 mmol; 2.0 eq.), cesium carbonate (1.66 g; 5.08 mmol; 2.5 eq.), Xphos (145.4 mg; 0.305 mmol; 0.15 eq.) and bis(acetonitrile) palladium(II) chloride (29.43 mg; 0.102 mmol; 0.05 eq.). The reaction mixture was purged with argon and stirred for 16 hours at 70°C. The reaction mixture was diluted with water (50 ml) and acidified with 1 N. aqueous hydrochloric acid to pH = 1. The aqueous layer was extracted with dichloromethane (3x100 ml) and the combined organic layers were dried over sodium sulfate, filtered and concentrated to dryness. The resulting residue was purified by silica gel column chromatography using cyclohexane/ethyl acetate as eluent to give the title compound (257 mg; 45%).

1H NMR (DMSO): 1.41 (s, 9H), 2.86 (s, 3H), 4.25 (s, 2H), 7.45 (d, 1H, J=11.6 Hz), 8.20 (d, 1H, J=11.2 Hz) , 10.87 (s, 1H); 13C NMR (DMSO): 28.01, 33.45, 37.80, 79.41, 82.10, 82.13, 83.56, 115.42, 115.53, 115.58, 133.79, 133.87, 140.96, 141.00, 142.56 , 142.65, 154.13, 154.47, 156.69; LC/MS [M-tBu] ⁺ : 225.02.

Step D: ethyl 2-[[6-(1,3-benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]methylamino]-5-[3-[[6-[3[tert-butoxycarbonyl(methyl )amino]prop-1-ynyl]-4-fluoro-3-pyridyl]oxy]propyl]thiazole-4carboxylate.

Using a general alkylation procedure, starting with the product of synthesis 5g and the product from step C as a suitable phenol, the title compound was obtained.

1H NMR (500 MHz, DMSO<) δ ppm 8.46 (d, 1H), 7.91 (br., 1H), 7.67 (s, 1H), 7.55 (d, 1H), 7.53 (br., 1H), 7.38 (t, 1H), 7.21 (t, 1H) , 4.28 (t, 2H), 4.25 (q, 2H), 4.24 (br., 2H), 3.76 (s, 3H), 3.27 (t, 2H), 2.85 (br., 3H), 2.46 (s, 3H ), 2.14 (m, 2H), 1.41 (s, 9H), 1.29 (t, 3H); ¹³ C NMR (125 MHz, DMSO<) δ ppm 138.8, 126.6, 122.6, 122.3, 118.4, 115.7, 69.3, 60.7, 38.4, 35.3, 33.9, 31.0, 28.4, 23.1, 17.8, 14.6; MSHR-ERI (m/z): [M+H] ⁺ , calculated for C ₃₆ H ₄₀ FN ₈ O ₅ S ₂ : 747.2541; found: 747.2526.

Step E: 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)-5-[3-({4fluoro-6-[ 3 -(methylamino)prop-1 -in-1-yl]pyridin-3 -yl}oxy)propyl] - 1,3-thiazol-4-carboxylic acid.

Using a general deprotection and hydrolysis procedure, starting with the product from Step D as a suitable ethyl ester, the desired product was obtained.

1H NMR (500 MHz, DMSO^6) δ ppm 8.44 (d, 1H), 7.91 (d, 1H), 7.67 (s, 1H), 7.53 (d, 1H), 7.48 (d, 1H), 7.39 (tm, 1H), 7.21 (tm, 1H), 4.27 (t, 2H), 3.76 (s, 3H), 3.48 (s, 2H), 3.27 (t, 2H), 2.46 (s, 3H), 2.31 (s, 3H), 2.14 (quin, 2H); ¹³ C NMR (125 MHz, DMSO<) δ ppm 164.1, 156.8, 156.7, 151.9, 151.9, 145.5, 143.8, 140.2, 138.8, 137.0, 136.0, 133.1, 130.3, 126.6, 122.6, 122.4, 118.4, 116. 8, 115.4, 88.2, 82.4, 69.4, 40.0, 35.3, 35.3, 30.9, 23.1, 17.7; MSHR-ERI (m/z): [M+H] ⁺ , calculated for C ₂ 9H ₂₈ FN ₈ O ₃ S ₂ : 619.1704; found: 619.1698.

Example 198: 2-[[6-(1,3-Benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]methylamino]-5-[3-[2-fluoro4-(3-morpholinoprop-1-ynyl )phenoxy]propyl]thiazole-4-carboxylic acid

- 236 046336

Step A: ethyl 5-[3-[2-fluoro-4-(3-morpholinoprop-1-ynyl)phenoxy]propyl]-2-[methyl-[5-methyl-6[^)-[3-(2 -trimethylsilylethoxymethyl)-1,3-benzothiazol-2-ylidene]amino]pyridazin-3-yl]amino]thiazol-4carboxylate.

Using a general alkylation procedure, starting with synthesis product 5g and synthesis product 6e as a suitable phenol, the desired product was obtained.

1H NMR (400 MHz, DMSOD6) δ ppm 7.84 (brd, 1H), 7.64 (d, 1H), 7.45 (dm, 1H), 7.42 (tm, 1H), 7.32 (dd, 1H), 7.25 (tm, 1H), 7.21 (dm, 1H), 7.15 (t, 1H), 5.85 (s, 2H), 4.25 (q, 2H), 4.15 (t, 2H), 3.77 (s, 3H), 3.71 (t, 2H), 3.58 (t, 4H), 3.44 ( s, 2H), 3.26 (t, 2H), 2.48 (br, 4H), 2.44 (s, 3H), 2.12 (quin, 2H), 1.29 (t, 3H), 0.82 (t, 2H), -0.11 ( s, 9H); ¹³ C NMR (100 MHz, DMSO-de) δ ppm 162.6, 157.4, 156.7, 155.1, 151.7, 151.6, 147.3, 141.0, 137.6, 137.0, 135.2, 129.0, 127.2, 125.6, 123.5, 123.2, 119.2, 117. 6, 115.4, 115.2, 111.9, 85.0, 84.3, 72.8, 68.4, 66.7, 66.5, 60.6, 52.2, 47.4, 35.3, 31.1, 23.3, 17.8, 17.8, 14.6, -1.0; MSHR-ERI (m/z): [M+H]+, calculated for C ₄₁ H ₅₁ FN ₇ O ₅ S ₂ Si: 832.3141; found: 832.3146.

Step B: 2-[[6-(1,3-benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]methylamino]-5-[3-[2-fluoro-4(3-morpholinoprop-1- inyl)phenoxy]propyl]thiazole-4-carboxylic acid.

Using a general deprotection and hydrolysis procedure, starting with the product from Step A as a suitable ethyl ester, the desired product was obtained.

MSVR-ERI (m/z): [M+H]+, calculated for C ₃₃ H33FN ₇ O ₄ S ₂ : 674.2014; found: 674.2006.

Example 199: 2-[[6-(1,3-Benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]-[6(dimethylamino)hexyl]amino]-5-[3-[2-fluoro- 4-[3-(methylamino)prop-1-ynyl]phenoxy]propyl]thiazole-4carboxylic acid /

Step A: methyl 2-[tert-butoxycarbonyl-[6-(dimethylamino)hexyl]amino]-5-[3-[4-[3-[tert-butoxycarbonyl(methyl)amino]prop-1-ynyl]-2-fluorophenoxy ]propyl]thiazole-4-carboxylate.

Using the general Mitsunobu reaction procedure, starting from the synthesis product 1b and 6(dimethylamino)hexan-1-ol, 1.51 g (amount containing approximately 40 mol/mol% DIAD-2H) of the desired product was obtained.

1H NMR (400 MHz, DMSO-de) δ ppm 7.30 (dd, 1H), 7.21 (dm, 1H), 7.13 (t, 1H), 4.23 (s, 2H), 4.10 (t, 2H), 3.98 (t, 2H), 3.75 (s, 3H), 3.22 (t, 2H), 2.86 (s, 3H), 2.13 (t, 2H), 2.07 (s, 6H), 1.70-1.20 (m, 8H), 1.61 (m, 2H), 1.50/1.41 (s, 18H ); MSHR-ERI (m/z): [M+H]+, calculated for C ₃₆ H ₅₄ FN ₄ O ₇ S: 705.3697; found: 705.3699.

Step B: methyl 5-[3-[4-[3-[tert-butoxycarbonyl(methyl)amino]prop-1-ynyl]-2fluorophenoxy]propyl]-2-[6-(dimethylamino)hexylamino]thiazol-4- carboxylate.

Using the general HFIP deprotection procedure, 650 mg (50%) of the desired product was obtained from the product from step A.

¹ H NMR (500 MHz, DMSOD6) δ ppm 7.56 (t, 1H), 7.31 (d, 1H), 7.21 (d, 1H), 7.13 (t, 1H), 4.23 (br., 2H), 4.07 (t, 2H), 3.69 (s, 3H), 3.14 (m, 2H), 3.11 (t, 2H), 2.86 (br., 3H), 2.17 (m, 2H), 2.10 (s, 6H), 2.00 (quint., 2H), 1.50 (m, 2H) , 1.41 (s, 9H), 1.37 (m, 2H), 1.34-1.21 (m, 2H), 1.34-1.21 (m, 2H); ¹³ C NMR (125 MHz, DMSO-de) δ ppm 129.1, 119.3, 115.4, 68.0, 60.6, 51.7, 45.6, 44.5, 38.6, 33.9, 30.6, 29.1, 28.5, 27.4, 27.0, 26.8, 23.3; MSHR-ERI (m/z): [M+H]+, calculated for C ₃₁ H ₄₆ FN ₄ O ₅ S: 605.3173; found: 605.3160.

Step C: methyl 5-[3-[4-[3-[tert-butoxycarbonyl(methyl)amino]prop-1-ynyl]-2

- 237 046336 fluorophenoxy]propyl]-2-[6-(dimethylamino)hexyl-[5-methyl-6-[(2)-[3-(2-trimethylsilylethoxymethyl)-1,3benzothiazol-2-ylidene] amino]pyridazine -3-yl]amino]thiazol-4-carboxylate.

Using the general procedure II for the Buchwald reaction, 1.06 g (98%) of the target product was obtained from the product from stage B and the product of synthesis 4a.

1H NMR (500 MHz, DMSO-de) δ ppm 7.84 (d, 1H), 7.66 (s, 1H), 7.47 (d, 1H), 7.44 (td, 1H), 7.32 (d, 1H), 7.25 (td, 1H), 7.22 (d, 1H), 7.16 (t, 1H), 5.86 (s, 2H), 4.37 (t, 2H), 4.20 (m, 2H), 4.15 (t, 2H), 3.77 (s, 3H), 3.72 (t, 2H), 3.26 ( t, 2H), 2.84 (s, 3H), 2.46 (s, 3H), 2.13 (m, 2H), 2.13 (m, 2H), 2.06 (s, 6H), 1.62-1.32 (m, 8H), 1.40 (s, 9H), 0.92 (t, 2H), -0.11 (s, 9H); ¹³ C NMR (125 MHz, DMSO^) δ ppm 163.2, 129.1, 127.2, 123.4, 123.2, 119.2, 117.6, 115.5, 112.0, 79.5, 72.8, 68.4, 66.7, 59.5, 51.9, 46.8, 45.6, 38.6, 33.8, 31.0, 28.5, 23.1, 17.8, 17.7, 0.96; MSHR-ERI (m/z): [M+H]+, calculated for C49H6 ₈ FN ₈ O ₆ S2Si: 975.4457; found: 975.4437.

Step D: 2-[[6-(1,3-benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]-[6(dimethylamino)hexyl]amino]-5-[3-[2-fluoro- 4-[3-(methylamino)prop-1-ynyl]phenoxy]propyl]thiazole-4carboxylic acid.

Using the general deprotection and hydrolysis procedure followed by repurification by reverse phase preparative chromatography (C18, 0.1% TFA in water:MeCN), the TFA salt of the desired product was obtained from the product from step C.

MSHR-ERI (m/z): [M+H]+, calculated for C ₃ 7H4 ₄ FN ₈ O ₃ S2: 731.2962; found: 731.2968.

Example 200: 2-[[6-(1,3-Benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]-[3(dimethylamino)propyl]amino]-5-[3-[2-fluoro- 4-[3-(methylamino)prop-1-ynyl]phenoxy]propyl]thiazole-4carboxylic acid

Step A: methyl 2-[tert-butoxycarbonyl-[3-(dimethylamino)propyl]amino]-5-[3-[4-[3-[tert-butoxycarbonyl(methyl)amino]prop-1-ynyl]-2-fluorophenoxy ]propyl]thiazole-4-carboxylate.

Using the general Mitsunobu reaction procedure, starting from the synthesis product 1b and 3(dimethylamino)propan-1-ol, 1.40 g (quantit., sample contained approximately 35 mol./mol.% DIAD-2H) of the desired product was obtained.

1H NMR (400 MHz, DMSO^ ₆ ) δ ppm 7.30 (dd, 1H), 7.21 (dm, 1H), 7.13 (t, 1H), 4.23 (s, 2H), 4.10 (t, 2H), 4.01 (t, 2H), 3.74 (s, 3H), 3.22 (t, 2H), 2.86 (s, 3H), 2.24 (t, 2H), 2.12 (s, 6H), 2.08 (m, 2H), 1.74 (m, 2h), 1.51/1.41 (s, 18H); MSHR-ERI (m/z): [M+H]+, calculated for C ₃₃ H ₄₈ FN ₄ O ₇ S: 663.3228; found: 663.3218.

Step B: methyl 5-[3-[4-[3-[tert-butoxycarbonyl(methyl)amino]prop-1-ynyl]-2fluorophenoxy]propyl]-2-[3-(dimethylamino)propylamino]thiazol-4- carboxylate.

Using the general HFIP deprotection procedure, 0.95 g (80%) of the desired product was obtained from the product from step A.

1H NMR (500 MHz, DMSO-de) δ ppm 7.57 (t, 1H), 7.31 (d, 1H), 7.21 (d, 1H), 7.13 (t, 1H), 4.23 (br., 2H), 4.07 (t, 2H), 3.69 (s, 3H), 3.17 (q, 2H), 3.12 (t, 2h), 2.86 (br., 3H), 2.24 (t, 2H), 2.11 (s, 6H), 2.00 (quint., 2h), 1.63 (m, 2H) , 1.41 (s, 9H); ¹³ C NMR (125 MHz, DMSOC) δ ppm 129.1, 119.3, 115.4, 68, 57.0, 51.7, 45.6, 42.8, 38.6, 33.8, 30.6, 28.5, 27.0, 23.3; MSHR-ERI (m/z): [M+H]+, calculated for C ₂₈ H4 ₀ FN ₄ O ₅ S: 563.2703; found: 563.2694.

Step C: methyl 5-[3-[4-[3-[tert-butoxycarbonyl(methyl)amino]prop-1-ynyl]-2-fluorophenoxy]propyl]-2-[3-(dimethylamino)propyl-[5 -methyl-6-[(2)-[3-(2-trimethylsilylethoxymethyl)-1,3-benzothiazol2-ylidene]amino]pyridazin-3-yl]amino]thiazol-4-carboxylate.

Using the general procedure II of the Buchwald reaction, starting from the product from stage B and the product of synthesis 4a, 0.79 g (51%) of the target product was obtained.

1H NMR (500 MHz, DMSO^) δ ppm 7.84 (d, 1H), 7.73 (s, 1H), 7.46 (dd, 1H), 7.43 (td, 1H), 7.31 (brd., 1H), 7.25 (td, 1H), 7.21 (d, 1H), 7.16 (t, 1H), 5.86 (s, 2H), 4.35 (t, 2H), 4.20 (br., 2H), 4.15 (t, 2H), 3.76 (s, 3H), 3.72 (t, 2H), 3.27 (t, 2H), 2.84 (br., 3H), 2.45 (s, 3H), 2.32 (t, 2H), 2.18 (s, 6H), 2.13 (m, 2H), 1.86 (m, 2H), 1.40 (s, 9H), 0.92 (t, 2H), -0.11 (s, 9H); ¹³ C NMR (125 MHz, DMSO^) δ ppm 129.1, 127.2, 123.4, 123.2, 119.3, 117.6, 115.4, 111.9, 72.8, 68.4, 66.7, 56.4, 51.9, 45.7, 45.5, 38.5, 33.8, 31.0, 28.5, 2 5.0, 23.1, 17.9, 17.8, -1.0; MSHR-ERI (m/z): [M+H]+, calculated for C46H6 ₂ FN8O6S ₂ Si: 933.3987; found: 933.3990.

Step D: 2-[[6-(1,3-benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]-[3-(dimethylamino)

- 238 046336 propyl]amino]-5-[3-[2-fluoro-4-[3-(methylamino)prop-1-ynyl]phenoxy]propyl]thiazole-4-carboxylic acid.

Using the general deprotection and hydrolysis procedure followed by repurification by reverse phase preparative chromatography (C18, 0.1% TFA in water:MeCN), the TFA salt of the desired product was obtained from the product from step C.

HRMS-ERI (m/z): [M+2H]2+, calculated for C ₃₄ H ₃ 9FN ₈ O ₃ S ₂ : 345.1280; found: 345.1265.

Example 201: 2-[[6-(1,3-Benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]-[2(trimethylammonio)ethyl]amino]-5-[3-[2-fluoro- 4-[3-(methylamino)prop-1-ynyl]phenoxy]propyl]thiazole-4carboxylate.

Using the general procedure of alkylation, deprotection and hydrolysis, starting from the compound of example 207, step C, and iodomethane, the TFA salt of the desired product was obtained.

MSHR-ERI (m/z): [M+2H] ²⁺ , calculated for C ₃₄ H ₃₉ FN ₈ O ₃ S ₂ : 345.1280; found: 345.1279.

Example 202: 2-[[6-(1,3-Benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]-[3(trimethylammonio)propyl]amino]-5-[3-[2-fluoro- 4-[3-(methylamino)prop-1-ynyl]phenoxy]propyl]thiazole 4-carboxylate.

Using the general procedure of alkylation, deprotection and hydrolysis, starting from the compound of Example 200, step C, and iodomethane, the TFA salt of the desired product was obtained.

MSHR-ERI (m/z): [M+2H] ²⁺ , calculated for C ₃₅ H ₄₁ FN ₈ O ₃ S ₂ : 352.1358; found: 352.1365.

Example 203: 2-[[6-(1,3-Benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]methylamino]-5-[3-[2-fluoro4-(3-pyrrolidin-1-ylprop -1-ynyl)phenoxy]propyl]thiazole-4-carboxylic acid

ABOUT

Step A: ethyl 5-[3-[2-fluoro-4-(3-pyrrolidin-1-ylprop-1-ynyl)phenoxy]propyl]-2-[methyl-[5methyl-6-[^)-[3 -(2-trimethylsilylethoxymethyl)-1,3-benzothiazol-2-ylidene]amino]pyridazin-3yl]amino]thiazol-4-carboxylate.

Using a general alkylation procedure, starting with synthesis product 5g and synthesis product 6c as a suitable phenol, the desired product was obtained.

1H NMR (400 MHz, DMSO-dg) δ ppm 7.83 (dm, 1H), 7.66 (q, 1H), 7.46 (dm, 1H), 7.43 (m, 1H), 7.31 (dd, 1H), 7.25 (m, 1H), 7.21 (dm, 1H), 7.15 (t, 1H), 5.85 (s, 2H), 4.25 (q, 2H), 4.14 (t, 2H), 3.77 (s, 3H), 3.72 (m, 2H), 3.64 (s, 2H), 3.26 ( t, 2H), 2.64 (brm, 4H), 2.45 (d, 3H), 2.12 (m, 2H), 1.72 (m, 4H), 1.29 (t, 3H), 0.92 (m,

- 239 046336

2H), -0.11 (s, 9H); MSHR-ERI (m/z): [M+H]+, calculated for C ₄₁ H ₅₁ FN ₇ O ₄ S ₂ Si: 816.3192; found: 816.3189.

Step B: 2-[[6-(1,3-benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]methylamino]-5-[3-[2-fluoro-4(3-pyrrolidin-1 - ilprop-1-ynyl)phenoxy]propyl]thiazole-4-carboxylic acid.

Using a general deprotection and hydrolysis procedure, starting with the product from Step A as a suitable ethyl ester, the desired product was obtained.

MSHR-ERI (m/z): [M+H]+, calculated for C ₃ 3H33FN ₇ O ₃ S ₂ : 658.2065; found: 658.2060.

Example 204: 2-[[6-(1,3-benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]methylamino]-5-[3-[2-fluoro4-[3-(1-piperidyl) prop-1-ynyl]phenoxy]propyl]thiazole-4-carboxylic acid

Step A: ethyl 5-[3-[2-fluoro-4-[3-(1-piperidyl)prop-1-ynyl]phenoxy]propyl]-2-[methyl-[5-methyl-6[^)- [3-(2-trimethylsilylethoxymethyl)-1,3-benzothiazol-2-ylidene]amino]pyridazin-3-yl]amino]thiazol-4carboxylate.

Using a general alkylation procedure, starting with synthesis product 5g and synthesis product 6d as a suitable phenol, the desired product was obtained.

1H NMR (400 MHz, DMSOD6) δ ppm 7.83 (dm, 1H), 7.64 (q, 1H), 7.45 (dm, 1H), 7.43 (m, 1H), 7.30 (dd, 1H), 7.25 (m, 1H), 7.19 (dm, 1H), 7.14 (t, 1H), 5.85 (s, 2H), 4.25 (q, 2H), 4.14 (t, 2H), 3.76 (s, 3H), 3.71 (m, 2H), 3.39 (s, 2H), 3.25 ( t, 2H), 2.44 (d, 3H), 2.42 (brm, 4H), 2.12 (m, 2H), 1.50 (m, 4H), 1.35 (m, 2H), 1.29 (t, 3H), 0.92 (m , 2H), -0.12 (s, 9H); MSHR-ERI (m/z): [M+H]+, calculated for C^^O^Si: 830.3348; found: 830.3347.

Step B: 2-[[6-(1,3-benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]methylamino]-5-[3-[2-fluoro-4[3-(1-piperidyl) )prop-1-ynyl]phenoxy]propyl]thiazole-4-carboxylic acid.

Using a general deprotection and hydrolysis procedure, starting with the product from Step A as a suitable ethyl ester, the desired product was obtained.

MSHR-ERI (m/z): [M+H]+, calculated for C ₃₄ H ₃₅ FN ₇ O ₃ S ₂ : 672.2221; found: 672.2217.

Example 205: 2-[[6-(1,3-benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]methylamino]-5-[3-[4-[3(dimethylamino)but-1-ynyl ]-2-fluorophenoxy]propyl]thiazole-4-carboxylic acid

Step A: ethyl 5-[3-[4-[3-(dimethylamino)but-1-ynyl]-2-fluorophenoxy]propyl]-2-[methyl-[5-methyl-6[^)-[3- (2-trimethylsilylethoxymethyl)-1,3-benzothiazol-2-ylidene]amino]pyridazin-3-yl]amino]thiazol-4carboxylate.

Using a general alkylation procedure, starting with synthesis product 5g and synthesis product 6f as a suitable phenol, the desired product was obtained.

Step B: 2-[[6-(1,3-benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]methylamino]-5 -[3-[4-[3(dimethylamino)but-1-ynyl ]-2-fluorophenoxy]propyl]thiazole-4-carboxylic acid.

Using a general deprotection and hydrolysis procedure, starting with the product from Step A as a suitable ethyl ester, the desired product was obtained.

MSHR-ERI (m/z): [M+H]+, calculated for C ₃₂ H ₃₃ FN ₇ O ₃ S ₂ : 646.2065; found: 646.2057.

Example 206: 2-[[6-(1,3-Benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]methylamino]-5-[3-[2-fluoro4-((3R or 3S)-morpholinobut -1-ynyl)phenoxy]propyl]thiazole-4-carboxylic acid (enantiomer 2).

- 240 046336

Step A: ethyl 5-[3-[2^TOp-4-((3R or 3S)-morpholinobut-1-ynyl)phenoxy]propyl]-2-[methyl-[5methyl-6-[^)-[3 -(2-trimethylsilylethoxymethyl)-1,3-benzothiazol-2-ylidene]amino]pyridazin-3yl]amino]thiazol-4-carboxylate (enantiomer 2).

Using a general alkylation procedure, starting with synthesis product 5g and synthesis product 6h as a suitable phenol, the desired product was obtained.

1HNMR (400 MHz, DMSO-de) δ ppm 7.84 (d, 1H), 7.66 (s, 1H), 7.46 (d, 1H), 7.43 (td, 1H), 7.29 (dd, 1H), 7.25 (td, 1H), 7.20 (dd, 1H), 7.15 (t, 1H), 5.86 (s, 2H), 4.26 (q, 2H), 4.14 (t, 2H), 3.77 (s, 3H), 3.72 (t, 2H), 3.66 (q, 1H), 3.59 ( br., 4H), 3.27 (t, 2H), 2.57/2.43 (m+m, 4H), 2.45 (s, 3H), 2.13 (quint., 2H), 1.30 (t, 3H), 1.28 (d, 3H), 0.92 (t, 2H), -0.11 (s, 9H); ¹³ C NMR (100 MHz, DMSO-de) δ ppm 129.0, 127.2, 123.4, 123.2, 119.2, 117.6, 115.5, 111.9, 88.1, 84.2, 72.8, 68.4, 66.7, 66.7, 60.7, 52.1, 49.5, 35.3, 31.1, 2 3.2, 19.3, 17.8, 17.8, 14.6, -1.0 ; HRMS-ERI (m/z): [M+H]+, calculated for C ₄₂ H ₅₃ FN ₇ O ₅ S ₂ Si: 846.3297; found: 846.3283.

Step B: 2-[[6-(1,3-benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]methylamino]-5-[3-[2-fluoro-4((3R or 3S)- morpholinobut-1-ynyl)phenoxy]propyl]thiazole-4-carboxylic acid (enantiomer 2).

Using a general deprotection and hydrolysis procedure, starting with the product from Step A as a suitable ethyl ester, the desired product was obtained.

MSHR-ERI (m/z): [M+H]+, calculated for C ₃₄ H ₃₅ FN ₇ O ₄ S ₂ : 688.2176; found: 688.2171.

Example 207: 2-[[6-(1,3-Benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]-[2(dimethylamino)ethyl]amino]-5-[3-[2-fluoro- 4-[3-(methylamino)prop-1-ynyl]phenoxy]propyl]thiazole-4carboxylic acid /

N

Step A: methyl 2-[tert-butoxycarbonyl-[2-(dimethylamino)ethyl]amino]-5-[3-[4-[3-[tert-butoxycarbonyl(methyl)amino]prop-1-ynyl]-2-fluorophenoxy ]propyl]thiazole-4-carboxylate.

Using the general Mitsunobu reaction procedure, 0.90 g (70%) of the target product was obtained from the synthesis product 1b and 2(dimethylamino)ethanol.

1H NMR (500 MHz, DMSOD6) δ ppm 7.30 (brd, 1H), 7.21 (dm, 1H), 7.13 (t, 1H), 4.22 (brs, 2H), 4.10 (t, 2H), 4.07 (t, 2H), 3.75 (s, 3H), 3.22 (t, 2H), 2.86 (brs, 3H), 2.46 (t, 2H), 2.17 (s, 6H), 2.08 (m, 2H), 1.50/1.41 (s, 18H); ¹³ C NMR (125 MHz, DMSO-de) δ ppm 129.1, 119.3, 115.4, 85.2, 82.4, 68.1, 57, 52.0, 46.0, 44.3, 38.6, 33.8, 30.5, 23.2; MSHR-ERI (m/z): [M+H]+, calculated for C ₃₂ H ₄₆ FN ₄ O ₇ S: 649.3071; found: 649.3059.

Step B: methyl 5-[3-[4-[3-[tert-butoxycarbonyl(methyl)amino]prop-1-ynyl]-2fluorophenoxy]propyl]-2-[2-(dimethylamino)ethylamino]thiazol-4- carboxylate.

Using the general HFIP deprotection procedure, 0.68 g (89%) of the desired product was obtained from the product from step A.

1H NMR (500 MHz, DMSO-dg) δ ppm 7.50 (t, 1H), 7.31 (d, 1H), 7.21 (d, 1H), 7.13 (t, 1H), 4.23 (br., 2H), 4.07 (t, 2H), 3.69 (s, 3H), 3.26 (q, 2H), 3.11 (t, 2H), 2.86 (br., 3H), 2.37 (t, 2H), 2.15 (s, 6H), 2.00 (quint., 2H), 1.41 (s, 9H) ; ¹³ C NMR (125 MHz, DMSO-de) δ ppm 129.1, 119.3, 115.4, 68.0, 58.2, 51.7, 45.7, 42.4, 38.6, 33.8, 30.6, 28.5, 23.3; MSHR-ERI (m/z): [M+H]+, calculated for C ₂₇ H ₃₈ FN ₄ O ₅ S: 549.2547; found: 549.2543.

Step C: methyl 5-[3-[4-[3-[tert-butoxycarbonyl(methyl)amino]prop-1-ynyl]-2fluorophenoxy]propyl]-2-[2-(dimethylamino)ethyl-[5-methyl -6-[^)-[3-(2-trimethylsilylethoxymethyl)-1,3benzothiazol-2-ylidene]amino]pyridazin-3-yl]amino]thiazol-4-carboxylate.

Using the general procedure II for carrying out the Buchwald reaction, starting from the product from stage B and

- 241 046336 of the synthesis product 4a obtained 0.57 g (50%) of the target product.

1H NMR (500 MHz, DMSO-dg) δ ppm 7.84 (d, 1H), 7.66 (s, 1H), 7.47 (dd, 1H), 7.44 (td, 1H), 7.31 (brd., 1H), 7.25 (t, 1H), 7.22 (d, 1H), 7.16 (t, 1H), 5.86 (s, 2H), 4.46 (t, 2H), 4.20 (br., 2H), 4.15 (t, 2H), 3.77 (s, 3H), 3.72 (t, 2H), 3.26 (t, 2H), 2.84 (br., 3H), 2.56 (t, 2H), 2.45 (s, 3H), 2.23 (s, 6H), 2.12 (m, 2H), 1.40 (s, 9H), 0.92 (t, 2H), -0.11 (s, 9H); ¹³ C NMR (125 MHz, DMSO-de) δ ppm 163.1, 129.1, 127.2, 123.5, 123.2, 119.3, 117.8, 115.4, 112.0, 72.9, 68.4, 66.7, 56.6, 51.9, 46.1, 45.3, 38.6, 33.8, 31.0, 28.5, 23.1, 17.9, 17.8, -1.0; MSHR-ERI (m/z): [M+H]+, calculated for C ₄₅ H ₆₀ FN ₈ O ₆ S ₂ Si: 919.3831; found: 919.3829.

Step D: 2-[[6-(1,3-benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]-[2(dimethylamino)ethyl]amino]-5-[3-[2-fluoro- 4-[3-(methylamino)prop-1-ynyl]phenoxy]propyl]thiazole-4carboxylic acid.

Using the general deprotection and hydrolysis procedure followed by repurification by reverse phase preparative chromatography (C18, 0.1% TFA in water:MeCN), the TFA salt of the desired product was obtained from the product from step C.

MSHR-ERI (m/z): [M+H]+, calculated for C ₃₃ H ₃₆ FN ₈ O ₃ S ₂ : 675.2336; found: 675.2323.

Example 208: 2-[[6-(1,3-Benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]methylamino]-5-[3-[2-fluoro4-((3S or 3R)-morpholinobut -1-ynyl)phenoxy]propyl]thiazole-4-carboxylic acid (enantiomer 1).

Step A: ethyl 5-[3-[2^mop-4-((3S or 3R)-morpholinobut-1-ynyl)phenoxy]propyl]-2-[methyl-[5methyl-6-[(7)-[ 3-(2-trimethylsilylethoxymethyl)-1,3-benzothiazol-2-ylidene]amino]pyridazin-3yl]amino]thiazol-4-carboxylate (enantiomer 1).

Using a general alkylation procedure, starting with synthesis product 5g and synthesis product 6g as a suitable phenol, the desired product was obtained.

1H NMR (400 MHz, DMSO-dg) δ ppm 7.84 (d, 1H), 7.66 (s, 1H), 7.46 (d, 1H), 7.43 (td, 1H), 7.29 (dd, 1H), 7.25 (td, 1H), 7.20 (dd, 1H), 7.15 (t, 1H), 5.86 (s, 2H), 4.26 (q, 2H), 4.14 (t, 2H), 3.77 (s, 3H), 3.72 (t, 2H), 3.66 (q, 1H), 3.59 ( br., 4H), 3.27 (t, 2H), 2.57/2.43 (m+m, 4H), 2.45 (s, 3H), 2.13 (quint., 2H), 1.30 (t, 3H), 1.28 (d, 3H), 0.92 (t, 2H), -0.11 (s, 9H); ¹³ C NMR (100 MHz, DMSO-dg) δ ppm 129.0, 127.2, 123.4, 123.2, 119.2, 117.6, 115.5, 111.9, 88.1, 84.2, 72.8, 68.4, 66.7, 66.7, 60.7, 52.1, 49.5, 35.3, 31.1, 2 3.2, 19.3, 17.8, 17.8, 14.6, -1.0 ; HRMS-ERI (m/z): [M+H]+, calculated for C ₄₂ H ₅₃ FN ₇ O ₅ S ₂ Si: 846.3297; found: 846.3288.

Step B: 2-[[6-(1,3-benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]methylamino]-5-[3-[2-fluoro-4((3S or 3R)- morpholinobut-1-ynyl)phenoxy]propyl]thiazole-4-carboxylic acid (enantiomer 1).

Using a general deprotection and hydrolysis procedure, starting with the product from Step A as a suitable ethyl ester, the desired product was obtained.

MSHR-ERI (m/z): [M+H]+, calculated for C ₃₄ H ₃₅ FN ₇ O ₄ S ₂ : 688.2170; found: 688.2166.

Example 209: 2-[[6-(1,3-benzothiazol-2-ylamino)-5-(trifluoromethyl)pyridazin-3-yl]methylamino]-5[3-[2-fluoro-4-[3-( methylamino)prop-1-ynyl]phenoxy]propyl]thiazole-4-carboxylic acid

Step A: methyl 2-[[6-chloro-5-(trifluoromethyl)pyridazin-3-yl]methylamino]-5-[3-(2-fluoro-4iodophenoxy)propyl]thiazole-4-carboxylate.

ml of an oven-dried, single-neck, round-bottomed flask was equipped with a PTFE-coated magnetic stir bar and a reflux condenser was installed. It was loaded with 991 mg of synthesis product 3s (2.20 mmol, 1.1 eq.), dissolved in 10 ml of dry THF, then the mixture was cooled to 0°C and 96 mg of NaH (2.40 mmol, 1.2 eq., 60% in mineral oil) was added in portions ) during the period

- 242 046336 min. After stirring for 15 minutes at 0°C, 434 mg of 3,6dichloro-4-(trifluoromethyl)pyridazine (2 mmol, 1 eq.) was added in one portion and stirred at this temperature for 1 hour, during which time the reaction reached a state of complete transformation. The reaction mixture was quenched by adding conc. NH ₄ Cl solution and extracted with EtOAc. The combined organic layers were dried over Na ₂ SO ₄ , filtered and concentrated under reduced pressure. The crude product was dissolved in DCM, then celite was added and the volatiles were removed under reduced pressure. The crude product was purified by flash column chromatography using heptane and EtOAc as eluents to obtain 500 mg (39%) of the title product as a yellow solid.

¹ H NMR (500 MHz, DMSOO δ ppm 8.16 (s, 1H), 7.58 (dd, 1H), 7.45 (m, 1H), 6.98 (t, 1H), 4.09 (t, 2H), 3.87 ( s, 3H), 3.80 (s, 3H), 3.28 (t, 2H), 2.11 (quin, 2H); ¹³ C NMR (125 MHz, DMSO-de) δ ppm 162.7, 156.2, 155.3, 152.2, MSVR-ERI (m/z): [M+H]+, calculated o for C ₂₀ H ₁₇ ClF ₄ IN ₄ O ₃ S: 630.9685; found: 630.9679.

Step B: methyl 5-[3-[4-[3-[tert-butoxycarbonyl(methyl)amino]prop-1-ynyl]-2fluorophenoxy]propyl]-2-[[6-chloro-5-(trifluoromethyl)pyridazine -3-yl]methylamino]thiazol-4-carboxylate.

A 12 ml oven-dried vessel equipped with a PTFE-coated magnetic stirrer arm was loaded with 330 mg of the product from step A (0.52 mmol, 1 eq.), 177 mg of tert-butyl N-methylNprop-2-ynylcarbamate (1.04 mmol, 2 eq.), 2.5 ml dry THF and 0.5 ml DIPA, and then created an inert atmosphere through the gas inlet. After adding 11.5 mg of Pd(PPh ₃ ) ₂ Cl ₂ (0.026 mmol, 0.05 eq.) and 4.98 mg of CuI (0.026 mmol, 0.05 eq.), the reaction mixture was heated to 60°C and stirred at this temperature for 1 hour, while during which time the reaction reached a state of complete transformation. Celite was added to the reaction mixture, and volatiles were removed under reduced pressure. The crude product was then purified by flash column chromatography using heptane and EtOAc to give 271 mg (69%) of the desired product.

1H NMR (500 MHz, DMSO-ph) δ ppm 8.15 (s, 1H), 7.31 (brd, 1H), 7.21 (dm, 1H), 7.14 (t, 1H), 4.23 (brs, 2H), 4.12 (t, 2H), 3.87 (s, 3H), 3.79 (s, 3H), 3.28 (t, 2H), 2.86 (brs, 3H), 2.12 (m, 2H), 1.41 (s, 9H); ¹³ C NMR (125 MHz, DMSO-de) δ ppm 144.2, 129.1, 121.6, 119.3, 117.8, 115.4, 85.2, 82.3, 68.2, 52.1, 38.6, 38.6, 35.8, 30.6, 28.5, 23.1; HRMS-ERI (m/z): [M+H]+, calculated for C ₂₉ H ₃₁ ClF ₄ N ₅ O ₅ S: 672.1665; found: 672.1651.

Step C: methyl 2-[[6-(1,3-benzothiazol-2-ylamino)-5-(trifluoromethyl)pyridazin-3-yl]methylamino]5-[3-[4-[3-[tert-butoxycarbonyl (methyl)amino]prop-1-ynyl]-2-fluorophenoxy]propyl]thiazole-4carboxylate.

A 4 ml oven-dried vessel equipped with a PTFE-coated magnetic stirrer arm was charged with 130 mg of the product from step B (0.19 mmol, 1 eq.), 87 mg of 1,3-benzothiazol-2amine (0.58 mmol, 3 eq. ) and 168 μl of DIPEA (125 mg, 0.96 mmol, 5 equiv.) suspended in 1 ml of CyOH, and then an inert atmosphere was created in it by flushing the vessel with argon. After 2 min of stirring in an inert atmosphere, 18 mg of Pd ₂ (dba) ₃ (0.019 mmol, 0.1 eq.) and 23 mg of XantPhos (0.038 mmol, 0.2 eq.) were added. The resulting mixture was then heated to 140°C and stirred at this temperature for 4 hours, during which time the reaction reached a state of complete conversion. The reaction mixture was diluted with 1 ml DCM and purified by flash column chromatography using heptane and EtOAc as eluents to obtain 85 mg (56%) of the desired product.

1H NMR (500 MHz, DMSO-ph) δ ppm 9.19 (br., 1H), 7.95 (s, 1H), 7.82 (d, 1H), 7.39 (t, 1H), 7.32 (d, 1H), 7.32 (d, 1H), 7.22 (d, 1H), 7.21 (t, 1H), 7.16 (t, 1H), 4.20 (br., 3H), 4.14 (t, 2H), 3.84 (s, 3H), 3.29 (t, 2H), 2.84 (br., 2H) , 2.13 (quint., 2H), 1.40 (s, 9H); ¹³ C NMR (125 MHz, DMSO-de) δ ppm 163.0, 160.0, 156.8, 151.7, 151.1, 141.7, 137.0, 135.1, 129.1, 127.3, 126.6, 123.2, 122.9, 122.7, 119.1, 116.4, 115.5, 112. 1, 68.3, 38.5, 35.6, 33.8, 30.9, 28.5, 23.1; MSHR-ERI (m/z): [M+H]+, calculated for C ₃ 6H ₃ 6F4N ₇ O5S2: 786.2150; found: 786.2148.

Step D: 2-[[6-(1,3-benzothiazol-2-ylamino)-5-(trifluoromethyl)pyridazin-3-yl]methylamino]-5-[3[2-fluoro-4-[3 -( methylamino)prop-1-ynyl]phenoxy]propyl]thiazole-4-carboxylic acid.

Using a general deprotection and hydrolysis procedure, starting with the product from Step C as the appropriate methyl ester, the desired product was obtained.

MSHR-ERI (m/z): [M+H]+, calculated for C ₃₀ H ₂₆ F ₄ N ₇ O ₃ S ₂ : 672.1469; found: 672.1458.

Example 210: 2-[[6-(1,3-benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]-[4 (dimethylamino)butyl]amino]-5-[3-[2-fluoro- 4-[3-(methylamino)prop-1-ynyl]phenoxy]propyl]thiazole-4carboxylic acid

- 243 046336

Step A: Methyl 2-[tert-butoxycarbonyl-[4-(dimethylamino)butyl]amino]-5-[3-(2-fluoro-4iodophenoxy)propyl]thiazole-4-carboxylate.

Using the general Mitsunobu reaction procedure, 401 mg (quantitative) of the target product was obtained from the synthesis product 1a and 4(dimethylamino)butan-1-ol.

¹ H NMR (400 MHz, DMSO-06) δ ppm 7.60 (dd, 1H), 7.47 (dm, 1H), 6.99 (t, 1H), 4.08 (t, 2H), 4.03 (t, 2H), 3.76 (s, 3H), 3.23 (t, 2H), 3.13 (t, 2H), 2.74 (s, 6H), 2.07 (m, 2H), 1.66 (m, 2H), 1.65 (m, 2H), 1.53 (s, 9H); ¹³ C NMR (100 MHz, DMSO-dt,) δ ppm 162.8, 152.7, 147.0, 143.6, 135.5, 134.1, 125.0, 117.7, 84.0, 82.4, 68.2, 56.8, 52.1, 46.0, 42.6, 30.5, 28.2, 25.2, 23.2, 2 1.7; MSHR-ERI (m/z): [M+H]+, calculated for C25H36FIN3O5S: 636.1399; found: 636.1400.

Step B: methyl 5-[3-[4-[3-[tert-butoxycarbonyl(methyl)amino]prop-1-ynyl]-2fluorophenoxy]propyl]-2-[4-(dimethylamino)butylamino]thiazol-4- carboxylate.

Using the general HFIP deprotection procedure, crude methyl 2-[4-(dimethylamino)butylamino]-5-[3-(2fluoro-4-iodophenoxy)propyl]thiazol-4- was obtained from the product from step A and after concentration. carboxylate (LC-MS-ESI (m/z): [M+H] ⁺ , calculated for C ₂₀ H ₂₈ FIN ₃ O ₃ S: 536.1; found: 536.2), which then, by using the General Sonogashira Reaction Procedure with tert -butyl Nmethyl-U-prop-2-ynylcarbamate as an alkyne reagent resulted in 240 mg (67%) of the desired product.

¹ H NMR (500 MHz, DMSO^) δ ppm 7.59 (t, 1H), 7.31 (d, 1H), 7.21 (d, 1H), 7.13 (t, 1H), 4.23 (br., 2H), 4.07 (t, 2H), 3.69 (s, 3H), 3.15 (q, 2H), 3.11 (t, 2H), 2.86 (br., 3H), 2.18 (t, 2H), 2.09 (s, 6H), 2.00 (quint., 2H), 1.50 (m, 2H) , 1.43 (m, 2H), 1.41 (s, 9H); ¹³ C NMR (125 MHz, DMSO^) δ ppm 129.1, 119.3, 115.4, 68.0, 59.2, 51.7, 45.6, 44.5, 38.6, 33.9, 30.6, 28.5, 27.0, 24.9, 23.3; MSHR-ERI (m/z): [M+H] ⁺ , calculated for C ₂₉ H ₄₂ FN ₄ O ₅ S: 577.2854; found: 577.2854.

Step C: methyl 5-[3-[4-[3-[tert-butoxycarbonyl(methyl)amino]prop-1-ynyl]-2fluorophenoxy]propyl]-2-[4-(dimethylamino)butyl-[5-methyl -6-[^)-[3-(2-trimethylsilylethoxymethyl)-1,3benzothiazol-2-ylidene]amino]pyridazin-3-yl]amino]thiazol-4-carboxylate.

Using the general Buchwald reaction procedure II, starting from 235 mg of the product from step B (0.41 mmol, 1 eq.) and 249 mg of the product of synthesis 4a (0.61 mmol, 1.5 eq.) as the appropriate halide, 353 mg (90%) were obtained target product.

¹ H NMR (500 MHz, DMSO^) δ ppm 7.84 (d, 1H), 7.73 (s, 1H), 7.47 (dm, 1H), 7.44 (t, 1H), 7.33 (brd, 1H), 7.26 (t, 1H), 7.22 (dm, 1H), 7.17 (t, 1H), 5.86 (s, 2H), 4.39 (br, 2H), 4.21 (br, 2H), 4.14 (t, 2H), 3.80 (s, 3H), 3.72 (t, 2H), 3.28 ( t, 2H), 3.13 (br, 2H), 2.85 (brs, 3H), 2.70 (s, 6H), 2.47 (s, 3H), 2.14 (quin, 2H), 1.75 (brm, 4H), 1.42 (s , 9H), 0.92 (t, 2H), -0.10 (s, 9H); ¹³ C NMR (125 MHz, DMSO^ ₆ ) δ ppm 163.1, 157.6, 155.3, 151.5, 147.6, 147.4, 141.5, 137.6, 137.5, 135.0, 129.1, 127.2, 123.5, 123.2, 119.4, 117.5, 115.5, 114. 8, 112.0, 85.3, 82.3, 72.9, 68.5, 66.9, 56.5, 52.1, 46.2, 42.7, 38.4, 34.0, 31.1, 28.4, 24.5/21.5, 23.2, 17.9, 17.8, -1.1; MSHR-ERI (m/z): [M+H] ⁺ , calculated for C^^O^Si: 947.4138; found: 947.4144.

Step D: 2-[[6-(1,3-benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]-[4(dimethylamino)butyl]amino]-5-[3-[2-fluoro- 4-[3-(methylamino)prop-1-ynyl]phenoxy]propyl]thiazole-4carboxylic acid.

Using the general deprotection and hydrolysis procedure followed by repurification by reverse phase preparative chromatography (C18, 0.1% TFA in water:MeCN), the TFA salt of the desired product was obtained from the product from step C.

MSVR-ERI (m/z): [M+H] ⁺ , calculated for C ₃₅ H ₄₀ FN ₈ O ₃ S ₂ : 703.2649; found: 703.2637.

Example 211: 2-[[6-(1,3-benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]-[4-hydroxy-5(trimethylammonio)pentyl]amino]-5-[3-[ 4-[3-(dimethylamino)prop-1-ynyl]-2fluorophenoxy]propyl]thiazole-4-carboxylate.

- 244 046336

Step A: [2-[tert-butyl(diphenyl)silyl]oxy-5-[[5-[3-[4-[3-(dimethylamino)prop-1-ynyl]-2fluorophenoxy]propyl]-4-methoxycarbonylthiazole -2-yl]-[5-methyl-6-[^)-[3-(2trimethylsilylethoxymethyl)-1,3-benzothiazol-2-ylidene]amino]pyridazin-3 yl]amino]pentyl]trimethylammonium.

Using the general procedure for the formation of a quaternary salt, starting from the synthesis product 5b and N,Ndimethylmethanamine as a suitable tertiary amine, the desired product was obtained.

Step B: 2-[[6-(1,3-benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]-[4-hydroxy-5(trimethylammonio)pentyl]amino]-5-[3-[ 4-[3-(dimethylamino)prop-1-ynyl]-2fluorophenoxy]propyl]thiazole-4-carboxylate.

Using the general quaternary salt deprotection procedure, starting from the quaternary salt product from Step A, the desired product was obtained.

MSHR-ERI (m/z): [M+H]+, calculated for C ₃₈ H ₄₆ FN ₈ O ₄ S ₂ : 761.3062; found: 761.3057.

Example 212: 3-[5-[[6-(1,3-benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]-[4-carboxy-5-[3-[2fluoro-4-[3 -(methylamino)prop-1-ynyl]phenoxy]propyl]thiazol-2yl]amino]pentyldimethylammonio]propane-1-sulfonate.

Using the general procedure of alkylation, deprotection and hydrolysis, starting from the compound of example 213, step C, and oxathiolane 2,2-dioxide, the TFA salt of the desired product was obtained.

MSHR-ERI (m/z): [M+H] ⁺ , calculated for C ₅₁ H ₇₂ FN ₈ O ₉ S ₃ Si: 839.2843; found: 839.2823.

Example 213: 2-[[6-(1,3-Benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]-[5(dimethylamino)pentyl]amino]-5-[3-[2-fluoro- 4-[3-(methylamino)prop-1-ynyl]phenoxy]propyl]thiazole-4carboxylic acid

Step A: methyl 2-[tert-butoxycarbonyl-[5-(dimethylamino)pentyl]amino]-5-[3-[4-[3-[tert-butoxycarbonyl(methyl)amino]prop-1-ynyl]-2-fluorophenoxy ]propyl]thiazole-4-carboxylate.

Using the general Mitsunobu reaction procedure, starting from the synthesis product 1b and 5(dimethylamino)pentan-1-ol, 1.47 g (quantitative) of the target product was obtained.

¹ H NMR (500 MHz, DMSOC) δ ppm 7.30 (d, 1H), 7.21 (d, 1H), 7.13 (t, 1H), 4.22 (m, 2H), 4.10 (t, 2H), 3.99 (t, 2H), 3.75 (s, 3H), 3.22 (m, 2H), 2.86 (bs, 3H), 2.35 (bs, 2H), 2.24 (bs, 6H), 2.08 (m, 2H), 1.62-1.13 (m, 6H), 1.51 (s, 9H), 1.42 (s, 9H); ¹³ C NMR (125 MHz, DMSOC) δ ppm 129.1, 119.2, 115.4, 85.2, 79.8, 68.1, 58.8, 52.0, 46.4, 45.0, 38.3, 33.8, 30.5, 28.5, 28.1, 23.2; MSVR-ERI (m/z): [M+H] ⁺ , calculated for

- 245 046336

C ₃₅ H ₅₂ FN ₄ O ₇ S: 691.3541; found: 691.3524.

Step B: methyl 5-[3-[4-[3-[tert-butoxycarbonyl(methyl)amino]prop-1-ynyl]-2fluorophenoxy]propyl]-2-[5-(dimethylamino)pentylamino]thiazol-4- carboxylate.

Using the general HFIP deprotection procedure, 0.66 g (52%) of the desired product was obtained from the product from step A.

¹ H NMR (500 MHz, DMSO-d^ δ ppm 7.58 (t, 1H), 7.32 (brd., 1H), 7.21 (d, 1H), 7.13 (t, 1H), 4.23 (br., 2H), 4.07 (t, 2H), 3.69 (s, 3H), 3.15 (m, 2H), 3.11 (t, 2H), 2.87 (br., 3H), 2.35 (br., 2H), 2.24 (brs ., 6H), 2.00 (quint, 2H), 1.51 (m, 2H), 1.44 (m, 2H), 1.41 (s, 9H), 1.30 (m, 2H); ¹³ C NMR (125 MHz, DMSO^) δ ppm 129.1, 119.3, 115.4, 68.0, 58.9, 51.7, 45.0, 44.4, 38.6, 33.8, 30.6, 28.9, 28.5, 26.4, 24.5, 23.3; MSHR-ERI (m/z): [M+H ]+, calculated for C ₃ oH ₄₄ FN ₄ 05S: 591.3016; found: 591.3005.

Step C: methyl 5-[3-[4-[3-[tert-butoxycarbonyl(methyl)amino]prop-1-ynyl]-2fluorophenoxy]propyl]-2-[5-(dimethylamino)pentyl-[5-methyl -6-[^)-[3-(2-trimethylsilylethoxymethyl)-1,3benzothiazol-2-ylidene]amino]pyridazin-3-yl]amino]thiazol-4-carboxylate.

Using the general procedure II of the Buchwald reaction, starting from the product from stage B and the product of synthesis 4a, 0.43 g (40%) of the target product was obtained.

^1H NMR (500 MHz, DMSO-d^ δ ppm 7.84 (d, 1H), 7.69 (s, 1H), 7.47 (d, 1H), 7.44 (td, 1H), 7.32 (brd., 1H ), 7.25 (td, 1H), 7.22 (d, 1H), 7.15 (t, 1H), 5.86 (s, 2H), 4.38 (t, 2H), 4.20 (br., 2H), 4.15 (t, 2H ), 3.78 (s, 3H), 3.72 (t, 2H), 3.26 (t, 2H), 2.84 (br., 3H), 2.57 (br., 2H), 2.46 (s, 3H), 2.39 (br. , 6H), 2.12 (m, 2H), 1.71 (m, 2H), 1.58 (m, 2H), 1.40 (s, 9H), 1.37 (m, 2H), 0.92 (t, 2H), -0.11 (s , 9H); ¹³ C NMR (125 MHz, DMSO-de) δ ppm 129.1, 127.2, 123.5, 123.2, 119.3, 117.6, 115.5, 112, 72.9, 68.4, 66.7, 58.2, 52.0, 46.7 , 44.2, 38.4, 33.8, 31.0, 28.5, 26.8, 25.4, 23.8, 23.2, 17.9, 17.8, -1.0; MSVR-ERI (m/z): [M+H]+, calculated for C ₄₈ H66FNsO6S2Si: 961.4300; found: 461.42 93 .

Step D: 2-[[6-(1,3-benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]-[5(dimethylamino)pentyl]amino]-5-[3-[2-fluoro- 4-[3-(methylamino)prop-1-ynyl]phenoxy]propyl]thiazole-4carboxylic acid.

Using the general deprotection and hydrolysis procedure followed by repurification by reverse phase preparative chromatography (C18, 0.1% TFA in water:MeCN), the TFA salt of the desired product was obtained from the product from step C.

MSHR-ERI (m/z): [M+H]+, calculated for C ₃₆ H ₄₂ FN ₈ O ₃ S ₂ : 717.2805; found: 717.2779.

Example 214: 2-[[6-(1,3-benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]methylamino]-5-[3-[4-[3(dimethylamino)propyl]-2- fluorophenoxy]propyl]thiazole-4-carboxylic acid

Step A: ethyl 5-[3-[4-[3-(dimethylamino)propyl]-2-fluorophenoxy]propyl]-2-[methyl-[5-methyl-6[(E)-[3-(2- trimethylsilylethoxymethyl)-1,3-benzothiazol-2-ylidene]amino]pyridazin-3-yl]amino]thiazol-4carboxylate.

A mixture of 59 mg of the compound of Example 50, Step A (0.075 mmol, 1 eq.), and 6 mg of 10% palladium on charcoal (0.076 eq.) in 5 ml of ethanol was maintained under a hydrogen pressure of 4 bar at RT. for 18 hours. After filtration through celite, the product was concentrated to give 59 mg (99%) of the expected product.

1H NMR (500 MHz, DMSO^ ₆ ) δ ppm 9.30 (brs, 1H), 7.84 (dm, 1H), 7.68 (q, 1H), 7.48 (dm, 1H), 7.44 (m, 1H), 7.25 (m, 1H), 7.13 (dd, 1H), 7.10 (m, 1H), 6.97 (dm, 1H), 4.27 (q, 2H), 4.10 (t, 2H), 3.78 (s, 3H), 3.72 (m, 2H), 3.27 (t, 2H), 2.99 ( m, 2H), 2.75 (d, 6H), 2.55 (t, 2H), 2.46 (d, 3H), 2.12 (m, 2H), 1.88 (m, 2H), 1.30 (t, 3H), 0.92 (m , 2H), -0.11 (s, 9H); MSHR-ERI (m/z): [M+H]+, calculated for C ₃ 9H5 ₃ FN ₇ O ₄ S2Si: 794.3354; found: 794.3356.

Step B: 2-[[6-(1,3-benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]methylamino]-5-[3-[4-[3(dimethylamino)propyl]-2- fluorophenoxy]propyl]thiazole-4-carboxylic acid.

To a mixture of 59 mg of the product from step A (0.076 mmol, 1 eq.) in 5 ml of 1,4-dioxane was added 1.5 ml of 4N. HCl solution (6.0 mmol) and stirred at room temperature. for 18 h. After adding 510 mg LiOHxH2O (12.14 mmol, 160 eq.), the mixture was stirred at 60°C for 2 h. After concentration, the crude product was purified by flash column chromatography using DCM and MeOH (containing 1.2% NH ₃ ) as eluents, and then using reverse phase preparative chromatography (C18, 25 mM NH4HCO3 in water:MeCN) to obtain 3 mg (6%) of the desired product.

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MSHR-ERI (m/z): [M+H]+, calculated for C ₃₁ H ₃₅ FN ₇ O ₃ S2: 636.2226; found: 636.2221.

Example 215: 2-[[6-(1,3-benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]-(4-hydroxy-5quinuclidin-1-ium-1-ylpentyl)amino]-5- [3-[4-[3-(dimethylamino)prop-1-ynyl]-2fluorophenoxy]propyl]thiazole-4-carboxylate.

Step A: methyl 2-[[4-[tert-butyl(diphenyl)silyl]oxy-5-quinuclidin-1-ium-1-ylpentyl]-[5-methyl6-[^)-[3-(2-trimethylsilylethoxymethyl )-1,3-benzothiazol-2-ylidene]amino]pyridazin-3-yl]amino]-5-[3[4-[3-(dimethylamino)prop-1-ynyl]-2-fluorophenoxy]propyl]thiazole -4-carboxylate.

Using the general procedure for the formation of a quaternary salt, starting from the synthesis product 5b and quinuclidine as a suitable tertiary amine, the desired product was obtained.

Step B: 2-[[6-(1,3-benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]-(4-hydroxy-5-quinuclidin-1ium-1-ylpentyl)amino]-5- [3-[4-[3-(dimethylamino)prop-1-ynyl]-2-fluorophenoxy]propyl]thiazole-4carboxylate.

Using the general quaternary salt deprotection procedure, starting from the quaternary salt product from Step A, the desired product was obtained.

MSHR-ERI (m/z): [M+2H] ²⁺ , calculated for C ₄₂ H ₅₁ FN ₈ O ₄ S ₂ : 407.1724; found: 407.1724.

Example 216: 2-[[5-(1-Aza-4-azoniabicyclo[2.2.2]octan-4-yl)-4-hydroxypentyl]-[6-(1,3benzothiazol-2-ylamino)-5-methylpyridazine -3-yl]amino]-5-[3-[4-[3-(dimethylamino)prop-1-ynyl]-2fluorophenoxy]propyl]thiazole-4-carboxylate.

Step A: methyl 2-[[5-(1-aza-4-azoniabicyclo[2.2.2]octan-4-yl)-4-[tert-butyl(diphenyl)silyl]oxypentyl]-[5-methyl-6-[ ^)-[3-(2-trimethylsilylethoxymethyl)-1,3-benzothiazol-2ylidene]amino]pyridazin-3-yl]amino]-5-[3-[4-[3-(dimethylamino)prop-1-ynyl ]-2fluorophenoxy]propyl]thiazole-4-carboxylate.

Using the general procedure for the formation of the quaternary salt, starting from the synthesis product 5b and 1,4diazabicyclo[2.2.2]octane as a suitable tertiary amine, the desired product was obtained.

Step B: 2-[[5-(1-aza-4-azoniabicyclo[2.2.2]octan-4-yl)-4-hydroxypentyl]-[6-(1,3-benzothiazol-2ylamino)-5-methylpyridazine -3-yl]amino]-5-[3-[4-[3-(dimethylamino)prop-1-ynyl]-2fluorophenoxy]propyl]thiazole-4-carboxylate.

Using the general quaternary salt deprotection procedure, starting from the quaternary salt product from Step A, the desired product was obtained.

MSHR-ERI (m/z): [m+2H] ²⁺ , calculated for C ₄₁ H ₅₀ FN9O ₄ S ₂ : 407.6700; found: 407.6699.

Example 217: 2-[[6-(1,3-Benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]-[4-hydroxy-5-(4methylmorpholin-4-ium-4-yl)pentyl] amino]-5-[3-[4-[3-(dimethylamino)prop-1-ynyl]-2fluorophenoxy]propyl]thiazole-4-carboxylate.

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Step A: methyl 2-[[4-[tert-butyl(diphenyl)silyl]oxy-5-(4-methylmorpholin-4-yl-4-yl)pentyl]-[5methyl-6-[^)-[3 -(2-trimethylsilylethoxymethyl)-1,3-benzothiazol-2-ylidene] amino]pyridazin-3 -yl] amino] 5-[3-[4-[3-(dimethylamino)prop-1-ynyl]-2- fluorophenoxy]propyl]thiazole-4-carboxylate.

Using the general procedure for the formation of a quaternary salt, starting from the synthesis product 5b and 4methylmorpholine as a suitable tertiary amine, the desired product was obtained.

Step B: 2-[[6-(1,3-benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]-[4-hydroxy-5-(4methylmorpholin-4-ium-4-yl)pentyl] amino]-5-[3-[4-[3-(dimethylamino)prop-1-ynyl]-2fluorophenoxy]propyl]thiazole-4-carboxylate.

Using the general quaternary salt deprotection procedure, starting from the quaternary salt product from Step A, the desired product was obtained.

MSHR-ERI (m/z): [M+H]+, calculated for C ₄₀ H ₄₈ FN ₈ O ₅ S ₂ : 803.3168; found: 803.3165.

Example 218: 3-[2-[3-[4-[3-[2-[[6-(1,3-Benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]methylamino]4-carboxythiazol- 5-yl]propoxy]-3-fluorophenyl]prop-2-ynylamino]ethyldimethylammonio]propane-1 sulfonate.

ABOUT

Step A: 3-[2-[tert-butoxycarbonyl-[3-[4-[3-[4-ethoxycarbonyl-2-[methyl-[5-methyl-6-[^)-[3-(2trimethylsilylethoxymethyl)- 1,3-benzothiazol-2-ylidene]amino]pyridazin-3-yl]amino]thiazol-5yl]propoxy]-3-fluorophenyl]prop-2-ynyl]amino]ethyldimethylammonio]propane-1-sulfonate.

159 mg of synthesis product 5h (0.17 mmol, 1 eq.) and 208 mg of oxathiolane 2,2-dioxide (1.70 mmol, 10 eq.) were mixed in acetonitrile (6 ml/mmol), and the reaction mixture was stirred at 60°C for 2.5 h, during which the conversion reached >90%. The mixture was concentrated in celite and purified by flash column chromatography using EtOAc and MeOH (1.2% NH3) as eluants to give 126 mg (70%) of the title product as a yellow solid.

¹ H NMR (500 MHz, DMSO-dg) δ ppm 7.84 (d, 1H), 7.67 (s, 1H), 7.46 (d, 1H), 7.44 (t, 1H), 7.35 (br., 1H), 7.25 (t, 1H), 7.25 (br., 1H) , 7.18 (t, 1H), 5.86 (s, 2H), 4.28 (br., 2H), 4.26 (q, 2H), 4.15 (t, 2H), 3.77 (s, 3H), 3.72 (t, 2H) , 3.70 (brt., 2H), 3.46 (br., 2H), 3.46 (br., 2H), 3.27 (t, 2H), 3.08 (brs., 6H), 2.46 (s, 3H), 2.46 (t , 2H), 2.13 (quint., 2H), 2.02 (br., 2H), 1.44 (brs., 9H), 1.29 (t, 3H), 0.92 (t, 2H), -0.11 (s, 9H); ¹³ C NMR (125 MHz, DMSOdg) δ ppm 154.4, 129.2, 127.2, 123.4, 123.2, 119.3, 117.6, 115.5, 111.9, 72.8, 68.4, 66.7, 63.3, 60.7, 59.9, 50.7, 48.0, 40.7, 37.8, 35.3, 31.1, 28.3, 23.2, 19.4, 17.8, 17.8, 14.6, -1.0; MSHR-ERI (m/z): [M+H] ⁺ , calculated for C ₄₉ H6 ₈ FN ₈ O ₉ S ₃ Si: 1055.4019; found: 1055.4026.

Step B: 3-[2-[3-[4-[3-[2-[[6-(1,3-benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]methylamino]-4carboxythiazol-5 -yl]propoxy]-3-fluorophenyl]prop-2-ynylamino]ethyldimethylammonio]propane-1sulfonate.

Using the general procedure of deprotection and hydrolysis followed by re-purification by reverse phase preparative chromatography (C18, 25 mM NH4HCO3 in water:MeC^, the TFA salt of the desired product was obtained from the product from step A.

MSHR-ERI (m/z): [M+H] ⁺ , calculated for C36H42FN8O6S3: 797.2368; found: 797.2343.

Example 219: 2-[[6-(1,3-Benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]-[5(trimethylammonio)pentyl]amino]-5-[3-[2-fluoro- 4-[3-(methylamino)prop-1-ynyl]phenoxy]propyl]thiazole 4-carboxylate.

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Using the general procedure of alkylation, deprotection and hydrolysis, starting from the compound of example 213, step C, and iodomethane, the TFA salt of the desired product was obtained.

MSHR-ERI (m/z): [M+H]+, calculated for C ₃₇ H ₄₄ FN8O ₃ S ₂ : 731.2956; found: 731.2967.

Example 220: 2-[[6-(1,3-Benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]-[4-hydroxy-5-(1methylpiperidin-1-yl-1-yl)pentyl] amino]-5-[3-[4-[3-(dimethylamino)prop-1-ynyl]-2fluorophenoxy]propyl]thiazole-4-carboxylate.

Step A: methyl 2-[[4-[tert-butyl(diphenyl)silyl]oxy-5-(1-methylpiperidin-1-yl-1-yl)pentyl]-[5methyl-6-[(7)-[ 3-(2-trimethylsilylethoxymethyl)-1,3-benzothiazol-2-ylidene]amino]pyridazin-3-yl]amino]5-[3-[4-[3-(dimethylamino)prop-1-ynyl]-2 -fluorophenoxy]propyl]thiazole-4-carboxylate.

Using the general procedure for the formation of a quaternary salt, starting from the synthesis product 5b and 1methylpiperidine as a suitable tertiary amine, the desired product was obtained.

Step B: 2-[[6-(1,3-benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]-[4-hydroxy-5-(1methylpiperidin-1-yl-1-yl)pentyl] amino]-5-[3-[4-[3-(dimethylamino)prop-1-ynyl]-2fluorophenoxy]propyl]thiazole-4-carboxylate.

Using the general quaternary salt deprotection procedure, starting from the quaternary salt product from Step A, the desired product was obtained.

MSVR-ERI (m/z): [M+H] ⁺ , calculated for C ₄₁ H ₅₀ FN8O ₄ S ₂ : 801.3375; found: 801.3378.

Example 221: 2-[[6-(1,3-benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]-[5-(1,4-dimethylpiperazin1-ium-1-yl)-4-hydroxypentyl ]amino]-5-[3-[4-[3-(dimethylamino)prop-1-ynyl]-2fluorophenoxy]propyl]thiazole-4-carboxylate.

Step A: methyl 2-[[4-[tert-butyl(diphenyl)silyl]oxy-5-(1,4-dimethylpiperazin-1-yl-1yl)pentyl]-[5-methyl-6-[(7) -[3-(2-trimethylsilylethoxymethyl)-1,3-benzothiazol-2ylidene]amino]pyridazin-3-yl]amino]-5-[3-[4-[3-(dimethylamino)prop-1-ynyl]- 2fluorophenoxy]propyl]thiazole-4-carboxylate.

Using the general procedure for the formation of a quaternary salt, starting from the synthesis product 5b and 1,4dimethylpiperazine as a suitable tertiary amine, the desired product was obtained.

Step B: 2-[[6-(1,3-benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]-[5-(1,4-dimethylpiperazin-1

- 249 046336 i-1 -yl)-4-hydroxypentyl]amino]-5 -[3-[4-[3 -(dimethylamino)prop-1-ynyl]-2fluorophenoxy]propyl]thiazole-4-carboxylate.

Using the general quaternary salt deprotection procedure, starting from the quaternary salt product from Step A, the desired product was obtained.

MSHR-ERI (m/z): [M+H]+, calculated for C ₄₁ H ₅₁ FN ₉ O ₄ S ₂ : 816.3484; found: 816.3489.

Example 222: 2-[3-[4-[3-[2-[[6-(1,3-benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]methylamino]-4carboxythiazol-5 -yl] propoxy]-3-fluorophenyl]prop-2-ynylamino]ethyltrimethylammonium

Step A: 2-[tert-butoxycarbonyl-[3-[4-[3-[4-ethoxycarbonyl-2-[methyl-[5-methyl-6-[^)-[3-(2trimethylsilylethoxymethyl)-1,3 -benzothiazol-2-ylidene]amino]pyridazin-3-yl]amino]thiazol-5yl]propoxy]-3-fluorophenyl]prop-2-ynyl]amino]ethyltrimethylammonium

200 mg of synthesis product 5h (0.21 mmol, 1 eq.) was dissolved in acetonitrile (4 ml/mmol), then 152 mg of iodomethane (1.07 mmol, 5 eq.) was added and stirred at room temperature. for 1.5 hours. The reaction did not proceed further, so at this moment the process was stopped, observing a partial transformation. The volatiles were evaporated and the crude mixture (containing synthesis product 5h and target product) was directly transferred to the next step.

LC-MS-ESI (m/z): [M] ⁺ , calculated for C ₄₇ H ₆₄ FN ₈ O ₆ S ₂ Si: 947.4; found: 947.4.

Step B: 2-[3-[4-[3-[2-[[6-(1,3-benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]methylamino]-4carboxythiazol-5 -yl] propoxy]-3-fluorophenyl]prop-2-ynylamino]ethyltrimethylammonium.

Using the general deprotection and hydrolysis procedure followed by repurification by reverse phase preparative chromatography (C18, 25 mM NH4HCO ₃ in water:MeCN), starting from the product from step A, the desired product and compound of Example 223 were separated and isolated. HRMS -ERI (m/z): [M+H]+, calculated for C ₃₄ H ₃₈ FN ₈ O ₃ S ₂ : 689.2487; found: 689.2481.

Example 223: 2-[[6-(1,3-benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]methylamino]-5-[3-[4-[3-[2(dimethylamino)ethylamino] prop-1-ynyl]-2-fluorophenoxy]propyl]thiazole-4-carboxylic acid

Example 223 was isolated as a by-product in the preparation of Example 222, Step B.

MSHR-ERI (m/z): [M+H]+, calculated for C ₃₃ H ₃₆ FN ₈ O ₃ S ₂ : 675.2330; found: 675.2324.

Example 224: 2-[[6-(1,3-benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]-[6(trimethylammonio)hexyl]amino]-5-[3-[2-fluoro- 4-[3-(methylamino)prop-1-ynyl]phenoxy]propyl]thiazole 4-carboxylate.

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Using the general procedure of alkylation, deprotection and hydrolysis, starting from the compound of example 199, step C, and iodomethane, the TFA salt of the desired product was obtained.

1H NMR (500 MHz, DMSO-06) δ ppm 7.90 (d, 1H), 7.64 (s, 1H), 7.53 (brd, 1H), 7.39 (t, 1H), 7.26 (dd, 1H), 7.21 (t, 1H), 7.18 (m, 1H), 7.15 (t, 1H), 4.37 (t, 2H), 4.13 (t, 2H), 3.44 (brs, 2H), 3.30 (t, 2H), 3.24 (t, 2H), 3.03 (s, 9H), 2.45 ( s, 3H), 2.31 (s, 3H), 2.11 (quin, 2H), 1.72-1.39 (m, 8H); ¹³ C NMR (125 MHz, DMSO-06) δ ppm 151.5, 147.1, 128.8, 126.6, 122.6, 122.3, 119.1, 118.4, 117.0, 115.5, 88.5, 82.0, 69.0, 65.8, 52.6, 46.6, 40.3, 35.4, 31.2, 26.6-22.2, 23.2, 18.0; MSHR-ERI (m/z): [M+H]+, calculated for C ₃₈ H ₄₄ FN ₈ O ₃ S ₂ : 745.3118; found: 745.3110.

Example 225: 2-[[6-(1,3-benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]-[5-[3 hydroxypropyl(dimethyl)ammonio]pentyl]amino]-5-[3 - [4-[3-(dimethylamino)prop-1-ynyl]-2fluorophenoxy]propyl]thiazole-4-carboxylate.

Step A: 5-[[5-[3-[4-[3-(dimethylamino)prop-1-ynyl]-2-fluorophenoxy]propyl]-4methoxycarbonylthiazol-2-yl] - [ 5 -methyl-6 - [ (Z)-[3-(2-trimethylsilylethoxymethyl)-1,3-benzothiazol-2ylidene]amino]pyridazin-3-yl]amino]pentyl-(3-hydroxypropyl)dimethylammonium.

Using a general alkylation procedure with an in situ formed tosylate followed by purification by reverse phase flash column chromatography (C18, 0.1% TFA in water:MeCN), starting from the synthesis product 5f and 3-(dimethylamino)propan-1-ol in as a suitable amine, the desired product was obtained.

1H NMR (500 MHz, DMSO-06) δ ppm 10.35 (brs, 1H), 7.84 (dm, 1H), 7.72 (s, 1H), 7.48 (dd, 1H), 7.48 (dm, 1H), 7.46 (m, 1H), 7.35 (dm, 1H), 7.26 (m, 1H), 7.24 (t, 1H), 5.87 (s, 2H), 4.79 (brs, 1H), 4.42 (t, 2H), 4.29 (s, 2H), 4.19 (t, 2H), 3.79 ( s, 3H), 3.73 (m, 2H), 3.46 (m, 2H), 3.30 (m, 2H), 3.27 (t, 2H), 3.25 (t, 2H), 3.00 (s, 6H), 2.86 (s , 6H), 2.47 (s, 3H), 2.14 (m, 2H), 1.8 (m, 2H), 1.76 (m, 2H), 1.75 (m, 2H), 1.38 (m, 2H), 0.92 (m, 2H), -0.98 (s, 9H); MSHR-ERI (m/z): [M] ⁺ , calculated for C ₄₇ H ₆₆ FN ₈ O ₅ S ₂ Si: 933.4346; found: 933.4342.

Step B: 2-[[6-(1,3-benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]-[5-[3 hydroxypropyl(dimethyl)ammonio]pentyl]amino]-5-[3 - [4-[3-(dimethylamino)prop-1-ynyl]-2fluorophenoxy]propyl]thiazole-4-carboxylate.

Using the general deprotection and hydrolysis procedure followed by repurification by reverse phase preparative chromatography (C18, 0.1% TFA in water:MeCN), the TFA salt of the desired product was obtained from the product from step A.

MSHR-ERI (m/z): [M+H]+, calculated for C ₄₀ H ₅₀ FN ₈ O ₄ S ₂ : 789.3375; found: 789.3376.

Example 226: 2-[[6-(1,3-benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]-[5-[2 hydroxyethyl(dimethyl)ammonio]pentyl] amino]-5-[3 - [4-[3-(dimethylamino)prop-1-ynyl]-2fluorophenoxy]propyl]thiazole-4-carboxylate.

Step A: 5-[[5-[3-[4-[3-(dimethylamino)prop-1-ynyl]-2-fluorophenoxy]propyl]-4methoxycarbonylthiazol-2-yl] - [ 5 -methyl-6 - [ (Z)-[3-(2-trimethylsilylethoxymethyl)-1,3-benzothiazol-2ylidene]amino]pyridazin-3-yl]amino]pentyl-(2-hydroxyethyl)dimethylammonium.

Using the general procedure of alkylation with the in situ generated tosylate followed by purification by reverse phase flash column chromatography (C18, 0.1% TFA in water:MeCN), starting from the synthesis product 5f and 2-(dimethylamino)ethanol as a suitable amine, received the target product.

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1H NMR (500 MHz, DMSO^) δ ppm 7.84 (d, 1H), 7.72 (s, 1H), 7.48 (d, 1H), 7.47 (dd, 1H), 7.45 (t, 1H), 7.35 (d, 1H), 7.26 (t, 1H), 7.24 (t, 1H), 5.87 (s, 2H), 5.29 (brt., 1H), 4.42 (t, 2H), 4.28 (s, 2H), 4.19 (t, 2H), 3.81 (brt., 2H), 3.79 (s, 3H), 3.73 (t, 2H), 3.36 (m, 2H), 3.31 (m, 2H), 3.27 (t, 2H), 3.05 (s, 6H), 2.86 (s, 6H), 2.47 (s, 3H), 2.14 (m, 2H), 1.76 (m, 2H), 1.76 (m, 2H), 1.37 (m, 2H), 0.92 (t, 2H), -0.10 (s, 9H); ¹³ C NMR (125 MHz, DMSOC) δ ppm 129.6, 127.3, 123.5, 123.2, 119.7, 117.6, 115.5, 112.0, 72.9, 68.6, 66.8, 65.1, 64.4, 55.4, 52.0, 51.2, 46.9, 46.4, 42.2, 3 1.1, 26.7, 23.2, 23.2, 21.9, 17.9, 17.8, -0.9; MSVR-ERI (m/z): [M] ⁺ , calculated for C46H64FN8O ₅ S2Si: 919.4189; found: 919.4192.

Step B: 2-[[6-(1,3-benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]-[5-[2hydroxyethyl(dimethyl)ammonio]pentyl]amino]-5-[3- [4-[3-(dimethylamino)prop-1-ynyl]-2fluorophenoxy]propyl]thiazole-4-carboxylate.

Using the general deprotection and hydrolysis procedure followed by repurification by reverse phase preparative chromatography (C18, 0.1% TFA in water:MeCN), the TFA salt of the desired product was obtained from the product from step A.

MSHR-ERI (m/z): [M+2H] ²⁺ , calculated for C ₃₉ H ₄₉ FN ₈ O ₄ S ₂ : 388.16455; found: 388.1639.

Example 227: 2-[[6-(1,3-Benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]-[5[carboxymethyl(dimethyl)ammonio]pentyl]amino]-5-[3 -[ 4-[3-(dimethylamino)prop-1-ynyl]-2fluorophenoxy]propyl]thiazole-4-carboxylate.

Step A: 5-[[5-[3-[4-[3-(dimethylamino)prop-1-ynyl]-2-fluorophenoxy]propyl]-4methoxycarbonylthiazol-2-yl]-[5-methyl-6-[ ^)-[3-(2-trimethylsilylethoxymethyl)-1,3-benzothiazol-2ylidene]amino]pyridazin-3-yl]amino]pentyl-(2-ethoxy-2-oxoethyl)dimethylammonium.

Using a general alkylation procedure with an in situ generated tosylate followed by purification by reverse phase flash column chromatography (C18, 0.1% TFA in water:MeCN) starting from synthesis 5f and ethyl 2-(dimethylamino)acetate as a suitable amine , the target product was obtained.

1H NMR (500 MHz, DMSO^) δ ppm 10.33 (brs, 1H), 7.84 (dm, 1H), 7.72 (s, 1H), 7.48 (dd, 1H), 7.48 (dm, 1H), 7.46 (m, 1H), 7.35 (dm, 1H), 7.27 (m, 1H), 7.24 (t, 1H), 5.87 (s, 2H), 4.42 (t, 2H), 4.38 (s, 2H), 4.29 (s, 2H), 4.21 (q, 2H), 4.19 ( t, 2H), 3.79 (s, 3H), 3.73 (m, 2H), 3.46 (m, 2H), 3.27 (t, 2H), 3.18 (s, 6H), 2.87 (s, 6H), 2.47 (s , 3H), 2.14 (m, 2H), 1.79 (m, 2H), 1.75 (m, 2H), 1.37 (m, 2H), 1.22 (t, 3H), 0.92 (m, 2H), -0.10(s , 9H); MSVR-ERI (m/z): [M] ⁺ , calculated for C48H66FN8O6S2Si: 961.4294; found: 961.4293.

Step B: 2-[[6-(1,3-benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]-[5[carboxymethyl(dimethyl)ammonio]pentyl]amino]-5-[3 -[ 4-[3-(dimethylamino)prop-1-ynyl]-2fluorophenoxy]propyl]thiazole-4-carboxylate.

Using the general deprotection and hydrolysis procedure followed by repurification by reverse phase preparative chromatography (C18, 0.1% TFA in water:MeCN), the TFA salt of the desired product was obtained from the product from step A.

MSHR-ERI (m/z): [M+H] ⁺ , calculated for C ₃₉ H ₄₆ FN ₈ O ₅ S ₂ : 789.3011; found: 789.2995.

Example 228: 3-[[5-[[6-(1,3-benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]-[4-carboxy-5-[3-[2fluoro-4-[ (E)-3-oxoprop-1-enyl]phenoxy]propyl]thiazol-2-yl]amino]-2hydroxypentyl]dimethylammonio]propane-1-sulfonate.

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Example 228 was isolated as a by-product from the preparation of Example 146.

MSHR-ERI (m/z): [M+H]+, calculated for C38H45FN7O8S3: 842.2470; found: 842.2494.

Example 229: 2-[[6-(1,3-Benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]-[5-[2hydroxyethyl(dimethyl)ammonio]pentyl]amino]-5-[3- [2-fluoro-4-[3-(methylamino)prop-1ynyl]phenoxy]propyl]thiazole-4-carboxylate.

Step A: 5-[[5-[3-[4-[3-[tert-butoxycarbonyl(methyl)amino]prop-1-ynyl]-2fluorophenoxy]propyl]-4-methoxycarbonylthiazol-2-yl]-[5 -methyl-6-[(2)-[3-(2trimethylsilylethoxymethyl)-1,3-benzothiazol-2-ylidene]amino]pyridazin-3-yl]amino]pentyl-(2hydroxyethyl)dimethylammonium.

Using the general procedure of alkylation with the in situ generated tosylate followed by purification by reverse phase flash column chromatography (C18, 0.1% TFA in water:MeCN), starting from the synthesis product 5e and 2-(dimethylamino)ethanol as a suitable amine, received the target product.

¹ H NMR (500 MHz, DMSO-06) δ ppm 7.84 (dm, 1H), 7.71 (s, 1H), 7.48 (dm, 1H), 7.45 (m, 1H), 7.33 (dd, 1H), 7.26 (m, 1H), 7.23 (dm, 1H), 7.17 (t, 1H), 5.87 (s, 2H), 5.28 (brs, 1H), 4.42 (t, 2H), 4.20 (brs, 2H), 4.16 (t, 2H), 3.81 (m, 2H), 3.78 ( s, 3H), 3.73 (m, 2H), 3.36 (t, 2H), 3.31 (t, 2H), 3.27 (t, 2H), 3.04 (s, 6H), 2.84 (brs, 3H), 2.46 (s , 3H), 2.13 (m, 2H), 1.77 (m, 2H), 1.75 (m, 2H), 1.41 (s, 9H), 1.37 (m, 2H), 0.92 (m, 2H), -0.11 (s , 9H); MSVR-ERI (m/z): [M] ⁺ , calculated for C ₅₀ H ₇₀ FN ₈ O7S ₂ Si: 1005.4557; found: 1005.4554.

Step B: 2-[[6-(1,3-benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]-[5-[2hydroxyethyl(dimethyl)ammonio]pentyl]amino]-5-[3- [2-fluoro-4-[3-(methylamino)prop-1ynyl]phenoxy]propyl]thiazole-4-carboxylate.

Using the general deprotection and hydrolysis procedure followed by repurification by reverse phase preparative chromatography (C18, 0.1% TFA in water:MeCN), the TFA salt of the desired product was obtained from the product from step A.

MSHR-ERI (m/z): [M+H]+, calculated for C ₃₈ H ₄₆ FN ₈ O ₄ S ₂ : 761.3062; found: 761.3057.

Example 230: 2-[[6-(1,3-Benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]-[5-[3hydroxypropyl(dimethyl)ammonio]pentyl]amino]-5-[3- [2-fluoro-4-[3-(methylamino)prop-1ynyl]phenoxy]propyl]thiazole-4-carboxylate.

Step A: 5-[[5-[3-[4-[3-[tert-butoxycarbonyl(methyl)amino]prop-1-ynyl]-2fluorophenoxy]propyl]-4-methoxycarbonylthiazol-2-yl]-[5 -methyl-6-[(2)-[3-(2trimethylsilylethoxymethyl)-1,3-benzothiazol-2-ylidene]amino]pyridazin-3-yl]amino]pentyl-(3hydroxypropyl)dimethylammonium.

Using a general alkylation procedure with the in situ formed tosylate followed by purification by reverse phase flash column chromatography (C18, 0.1% TFA in water:MeCN), starting from the synthesis product 5e and 3-(dimethylamino)propan-1-ol in as a suitable amine, the desired product was obtained.

¹ H NMR (500 MHz, DMSO-06) δ ppm 7.84 (dm, 1H), 7.71 (s, 1H), 7.48 (dm, 1H), 7.45 (m, 1H), 7.33 (dd, 1H), 7.26 (m, 1H), 7.23 (dm, 1H), 7.18 (t, 1H), 5.87 (s, 2H), 4.78 (brs, 1H), 4.42 (t, 2H), 4.21 (s, 2H), 4.15 (t, 2H), 3.78 (s, 3H), 3.72 ( m, 2H), 3.46 (m, 2H), 3.30 (t, 2H), 3.27 (t, 2H), 3.25 (t, 2H), 2.99 (s, 6H), 2.85 (brs, 3H), 2.47 (s , 3H), 2.12 (m, 2H), 1.80 (m, 2H), 1.76 (m, 2H), 1.76 (m, 2H), 1.41 (s, 9H), 1.38 (m, 2H), 0.92 (m, 2H),

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0.11 (s, 9H); MSVR-ERI (m/z): [M] ⁺ , calculated for C51H ₇₂ FN8O ₇ S ₂ Si: 1019.4713; found: 1019.4713.

Step B: 2-[[6-(1,3-benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]-[5-[3hydroxypropyl(dimethyl)ammonio]pentyl]amino]-5-[3- [2-fluoro-4-[3-(methylamino)prop-1 inyl]phenoxy]propyl]thiazole-4-carboxylate.

Using the general deprotection and hydrolysis procedure followed by repurification by reverse phase preparative chromatography (C18, 0.1% TFA in water:MeCN), the TFA salt of the desired product was obtained from the product from step A.

MSHR-ERI (m/z): [M+H] ⁺ , calculated for C ₃₉ H ₄₈ FN ₈ O ₄ S ₂ : 775.3218; found: 775.3213.

Example 231: 2-[[6-(1,3-Benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]-[5[carboxymethyl(dimethyl)ammonio]pentyl]amino]-5-[3-[ 2-fluoro-4-[3-(methylamino)prop-1ynyl]phenoxy]propyl]thiazole-4-carboxylate.

Step A: 5-[[5-[3-[4-[3-[tert-butoxycarbonyl(methyl)amino]prop-1-ynyl]-2fluorophenoxy]propyl]-4-methoxycarbonylthiazol-2-yl]-[5 -methyl-6-[(2)-[3-(2trimethylsilylethoxymethyl)-1,3-benzothiazol-2-ylidene]amino]pyridazin-3-yl]amino]pentyl-(2-ethoxy2-oxoethyl)dimethylammonium.

Using a general alkylation procedure with an in situ generated tosylate followed by purification by reverse phase flash column chromatography (C18, 0.1% TFA in water:MeCN) starting from synthesis 5e and ethyl 2-(dimethylamino)acetate as a suitable amine , the target product was obtained.

¹ H NMR (500 MHz, DMSOHO δ ppm 7.84 (dm, 1H), 7.71 (q, 1H), 7.48 (dm, 1H), 7.44 (m, 1H), 7.33 (dd, 1H), 7.26 ( m, 1H), 7.23 (dm, 1H), 7.17 (t, 1H), 5.87 (s, 2H), 4.42 (t, 2H), 4.37 (s, 2H), 4.21 (q, 2H), 4.21 (brs , 2H), 4.16 (t, 2H), 3.78 (s, 3H), 3.72 (m, 2H), 3.46 (m, 2H), 3.27 (t, 2H), 3.18 (s, 6H), 2.84 (brs, 3H), 2.47 (d, 3H), 2.12 (m, 2H), 1.79 (m, 2H), 1.75 (m, 2H), 1.41 (s, 9H), 1.37 (m, 2H), 1.22 (t, 3H ), 0.92 (m, 2H), -0.10 (s, 9H); HRMS-ERI (m/z): [M] ⁺ , calculated for C5 ₂ H ₇₂ FN8O8S ₂ Si: 1047.4662; found: 1047.468.

Step B: 2-[[6-(1,3-benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]-[5[carboxymethyl(dimethyl)ammonio]pentyl]amino]-5-[3-[ 2-fluoro-4-[3-(methylamino)prop-1ynyl]phenoxy]propyl]thiazole-4-carboxylate.

Using the general deprotection and hydrolysis procedure followed by repurification by reverse phase preparative chromatography (C18, 0.1% TFA in water:MeCN), the TFA salt of the desired product was obtained from the product from step A.

MSHR-ERI (m/z): [M+H] ⁺ , calculated for C ₃ 8H44FN8O5S ₂ : 775.2855; found: 775.2857.

Example 232: 2-[[6-(1,3-Benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]-[4(trimethylammonio)butyl]amino]-5-[3-[2-fluoro- 4-[3-(methylamino)prop-1-ynyl]phenoxy]propyl]thiazole-4carboxylate.

Step A: 4-[[5-[3-[4-[3-[tert-butoxycarbonyl(methyl)amino]prop-1-ynyl]-2fluorophenoxy]propyl]-4-methoxycarbonylthiazol-2-yl]-[5 -methyl-6-[(2)-[3-(2trimethylsilylethoxymethyl)-1,3-benzothiazol-2-ylidene]amino]pyridazin-3yl]amino]butyltrimethylammonium.

Using the general alkylation procedure with the tosylate formed in situ, starting from the synthesis product 5c and \,\-dimethylmethanamine as a suitable amine, the TFA salt of the target product was obtained

- 254 046336 ta.

1H NMR (500 MHz, DMSO-de) δ ppm 7.84 (d, 1H), 7.71 (s, 1H), 7.48 (d, 1H), 7.45 (t, 1H), 7.33 (brd, 1H), 7.26 (t, 1H), 7.22 (dm, 1H), 7.17 (t, 1H), 5.87 (s, 2H), 4.40 (t, 2), 4.20 (brs, 2H), 4.15 (t, 2H), 3.79 (s, 3H), 3.73 (t, 2H), 3.46 ( t, 2H), 3.27 (t, 2H), 3.06 (s, 9H), 2.84 (brs, 3H), 2.46 (s, 3H), 2.12 (m, 2H), 1.82 (m, 2H), 1.75 (m , 2H), 1.40 (s, 9H), 0.92 (t, 2H), -0.10 (s, 9H); ¹³ C NMR (125 MHz, DMSO-de) δ ppm 129.1, 127.2, 123.6, 123.2, 119.3, 117.5, 115.5, 112.1, 72.9, 68.4, 66.7, 65.4, 52.8, 52.1, 46.3, 38.8, 33.8, 31.0, 28.5, 2 4.4, 23.3, 19.8, 18.0, 17.8, -1.0 ; MSHR-ERI (m/z): [M] ⁺ , calculated for C48H66FN8OgS2Si: 961.4294; found: 961.4312.

Step B: 2-[[6-(1,3-benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]-[4(trimethylammonio)butyl]amino]-5-[3-[2-fluoro- 4-[3-(methylamino)prop-1-ynyl]phenoxy]propyl]thiazole-4carboxylate.

Using the general deprotection and hydrolysis procedure followed by repurification by reverse phase preparative chromatography (C18, 0.1% TFA in water:MeCN), the TFA salt of the desired product was obtained from the product from step A.

MSVR-ERI (m/z): [M] ⁺ , calculated for C ₃₆ H ₄₂ FN ₈ O ₃ S ₂ : 717.2800; found: 717.2800.

Example 233: 2-[[6-(1,3-benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]methylamino]-5-[3-[4-[3(dimethylamino)prop-1-ynyl ]phenoxy]propyl]thiazole-4-carboxylic acid

ABOUT

Step A: ethyl 5-[3-[4-[3-(dimethylamino)prop-1-ynyl]phenoxy]propyl]-2-[methyl-[5-methyl-6-[^)[3-(2- trimethylsilylethoxymethyl)-1,3-benzothiazol-2-ylidene]amino]pyridazin-3-yl]amino]thiazol-4carboxylate.

Using a general procedure for the alkylation of silyl-protected phenols starting from 219 mg of synthesis 5g (0.30 mmol, 1 eq) and 100 mg of synthesis 6j (0.30 mmol, 1 eq) followed by purification by flash column chromatography using EtOAc and MeOH (containing 1.2% NH3) as eluents gave 37 mg (12%) of the desired product as a dark yellow sticky compound.

1H NMR (500 MHz, DMSO-dg) δ ppm 7.83 (dm, 1H), 7.66 (q, 1H), 7.47 (dm, 1H), 7.44 (m, 1H), 7.35 (m, 2H), 7.25 (m, 1H), 6.94 (m, 2H), 5.86 (s, 2H), 4.26 (q, 2H), 4.05 (t, 2H), 3.77 (s, 3H), 3.72 (m, 2H), 3.38 (s, 2H), 3.26 (m, 2H), 2.45 ( d, 3H), 2.19 (s, 6H), 2.10 (m, 2H), 1.30 (t, 3H), 0.92 (m, 2H), -0.11 (s, 9H); MSHR-ERI (m/z): [M+H] ⁺ , calculated for C ₃₉ H ₅₀ N ₇ O ₄ S ₂ Si: 772.3129; found: 772.3119.

Step B: 2-[[6-(1,3-benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]methylamino]-5-[3-[4-[3(dimethylamino)prop-1-ynyl ]phenoxy]propyl]thiazole-4-carboxylic acid.

Using the general deprotection and hydrolysis procedure followed by repurification by reverse phase preparative chromatography (C18, 0.1% TFA in water:MeCN), the TFA salt of the desired product was obtained from the product from step A.

MSHR-ERI (m/z): [M+H] ⁺ , calculated for C ₃₁ H ₃₂ N ₇ O ₃ S ₂ : 614.2002; found: 614.1998.

Example 234: 2-[[6-(1,3-benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]methylamino]-5-[3-[4-[1[(dimethylamino)methyl]-3 -bicyclo[1.1.1]pentanyl]phenoxy]propyl]thiazole-4-carboxylic acid

Step A: ethyl 5-[3-[4-[1-[(dimethylamino)methyl]-3-bicyclo[1.1.1]pentanyl]phenoxy]propyl]-2[methyl-[5-methyl-6-[^ )-[3-(2-trimethylsilylethoxymethyl)-1,3-benzothiazol-2-ylidene]amino]pyridazin-3yl]amino]thiazol-4-carboxylate.

Using a general alkylation procedure starting from 47 mg of synthesis 6i (0.22 mmol, 1 eq.) and 157 mg of synthesis product 5g (0.22 mmol, 1 eq.) followed by column purification

- 255 046336 flash chromatography using EtOAc and MeOH (containing 1.2% NH3) as eluents gave 130 mg (74%) of the title product as a yellow solid.

¹ H NMR (500 MHz, DMSO-d,) δ ppm 10.62 (brs, 1H), 7.83 (dm, 1H), 7.65 (q, 1H), 7.46 (dm, 1H), 7.43 (m, 1H), 7.25 (m, 1H), 7.11 (m, 2H), 6.89 (m, 2H), 5.85 (s, 2H), 4.26 (q, 2H), 4.01 (t, 2H), 3.76 (s, 3H), 3.72 (m, 2H), 3.24 (s, 2H), 3.24 ( t, 2H), 2.74 (s, 6H), 2.44 (d, 3H), 2.08 (m, 2H), 2.07 (s, 6H), 1.29 (t, 3H), 0.91 (m, 2H), 0.12 (s , 9Н); HRMS-ERI (m/z): [M+H]+, calculated for C ₄₂ H ₅₆ N ₇ O ₄ S ₂ Si: 814.3599; found: 814.3592.

Step B: 2-[[6-(1,3-benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]methylamino]-5-[3-[4-[1[(dimethylamino)methyl]-3 -bicyclo[1.1.1]pentanyl]phenoxy]propyl]thiazole-4-carboxylic acid.

Using the general deprotection and hydrolysis procedure followed by repurification by reverse phase preparative chromatography (C18, 0.1% TFA in water:MeCN), the TFA salt of the desired product was obtained from the product from step A.

MSHR-ERI (m/z): [M+H] ⁺ , calculated for C ₃₄ H ₃₈ N ₇ O ₃ S ₂ : 656.2472; found: 656.2467.

Example 235: 3-[3-[[6-(1,3-benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]-[4-carboxy-5-[3-[2fluoro-4-[3 -(methylamino)prop-1-ynyl]phenoxy]propyl]thiazol-2yl]amino]propyldimethylammonio]propane-1-sulfonate.

The general alkylation, deprotection and hydrolysis procedure was used starting from Example 200, Step C, and oxathiolane 2,2-dioxide. After alkylation, the sulfonated intermediate was isolated (MSHR-ESI (m/z): [M+H] ⁺ , calculated for C49H68FN8O9S3Si: 1055.4025; found: 1055.4028), which was then reacted with 111 ^; chpyridine (HRMS-ERI (m/z): [M+H] ⁺ , calculated for C38H ₄₅ FN ₈ O ₆ S ₃ : 825.2686; found: 825.2675) and finally hydrolyzed with

LiOHxH ₂ O. Purification led to the TFA salt of the target product.

MSVR-ERI (m/z): [M+H] ⁺ , calculated for C ₃₇ H ₄₄ FN ₈ O ₆ S ₃ : 811.2530; found: 811.2525.

Example 236: 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl} (methyl)amino)-5-[3-(2fluoro-4-{3 -[methyl({4-[3-(trifluoromethyl)-3H-diazirin-3-yl]phenyl}methyl)amino]prop-1-in-1yl}phenoxy)propyl]-1,3-thiazol-4-carboxylic acid

I o

Step A: ethyl 5-[3-(2-fluoro-4-{3-[methyl({4-[3-(trifluoromethyl)-3H-diazirin-3yl]phenyl}methyl)amino]prop-1-in- 1-yl}phenoxy)propyl]-2-[methyl(5-methyl-6-{[(2Z)-3-{[2(trimethylsilyl)ethoxy]methyl }-2,3-dihydro-1,3-benzothiazole -2-ylidene] amino } pyridazin-3 -yl) amino] 1,3-thiazol-4-carboxylate.

To a solution of synthesis product 5j (20 mg, 25.8 µmol, 1 equiv.) in dry dimethylformamide (1 ml) was added potassium carbonate (3.9 mg, 28.4 µmol, 1.1 equiv.) followed by the addition of 3-[4(bromomethyl)phenyl]- 3-(trifluoromethyl)diazirine (7.9 mg, 28.4 μmol, 1.1 eq.), and the mixture was stirred for 16 h at ambient temperature. The reaction mixture was diluted with 5% aqueous ammonium hydroxide (10 ml), extracted with ethyl acetate (2x10 ml) and the combined organic extracts washed successively with 5% aqueous ammonium hydrofluoride (10 ml) and brine (10 ml), dried (magnesium sulfate) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 4 g RediSep™ silica gel cartridge) eluting with a gradient of 0-50% ethyl acetate in isoheptane gave the title product as a white solid (27.4 mg), which was used directly next stage without further characterization.

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Step B: ethyl 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)-5-[3-(2fluoro-4-{ 3-[methyl({4-[3-(trifluoromethyl)-3H-diazirin-3-yl]phenyl}methyl)amino]prop-1-in-1yl}phenoxy)propyl]-1,3-thiazol-4- carboxylate.

To a solution of the product from step A (27.4 mg, 28 µmol, 1 eq.) in acetonitrile (1 ml) was added polyhydrogen pyridinium fluoride (101 µl, 0.78 mmol, 28 eq.), and the mixture was heated at 60°C for 20 h The reaction mixture was diluted with 5% aqueous ammonium hydroxide (10 ml), extracted with ethyl acetate (2x10 ml) and the combined organic extracts were dried (magnesium sulfate) and concentrated in vacuo. Purification by reverse phase automated flash chromatography (CombiFlash Rf, C18 5.5 g RediSep aq Gold column) eluting with a gradient of 5-95% acetonitrile in water gave the title product as a yellow solid (6.7 mg, 0.01 mmol, 28%).

LC/MS (C _4i H ₃₇ F ₄ N ₉ O ₃ S ₂ ) 844 [M+H]+; RT 1.64 (LCMS-V-BI).

Step C: 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)-5-[3-(2-fluoro4-{3 -[methyl({4-[3-(trifluoromethyl)-3H-diazirin-3-yl]phenyl}methyl)amino]nrop-1-in-1yl}phenoxy)propyl]-1,3-thiazol-4-carboxylic acid.

To a solution of the product from step B (6.7 mg, 0.01 mmol, 1 eq.) in a 1:1 mixture of tetrahydrofuran/methanol (0.5 ml) was added a 2 M aqueous solution of sodium hydroxide (79.4 μl, 0.16 mmol, 20 eq.), and the mixture stirred at ambient temperature for 4 hours, and then concentrated in vacuo. The residue was diluted with water (0.2 ml) and adjusted to pH 5 with 2 M aqueous hydrochloric acid solution. Purification by reverse phase automated flash chromatography (CombiFlash Rf, C18 5.5 g RediSep aq Gold column) eluting with a gradient of 5-95% acetonitrile in water gave the title product as an orange/brown solid (3.5 mg, 4.29 µmol, 54%).

LC/MS (C ₃₉ H ₃₃ F ₄ N ₉ O ₃ S ₂ ) 816 [M+H] ⁺ ; RT 1.53 (LCMS-V-BI).

Example 237: 2-[[6-(1,3-benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]-(3-phosphonopropyl)amino]5-[3-[2-fluoro-4-[ 3-(methylamino)prop-1-ynyl]phenoxy]propyl]thiazole-4-carboxylic acid

HN

Step A: Methyl 2-[tert-butoxycarbonyl(3-diethoxyphosphorylpropyl)amino]-5-[3-[4-[3-[tert-butoxycarbonyl(methyl)amino]prop-1-ynyl]-2-fluorophenoxy]propyl]thiazole -4-carboxylate.

Using the general alkylation procedure, starting with 500 mg of synthesis 1b (0.87 mmol, 1 eq.) and 336 mg of 1-bromo-3-diethoxyphosphorylpropane (1.30 mmol, 1.5 eq.) as the appropriate halide, gave 517 mg (79%) target product.

¹ H NMR (500 MHz, DMSOC) δ ppm 7.30 (dd, 1H), 7.21 (dm, 1H), 7.13 (t, 1H), 4.23 (brs, 2H), 4.10 (t, 2H), 4.04 (t, 2H), 4.03-3.90 (m, 4H) , 3.75 (s, 3H), 3.22 (t, 2H), 2.86 (brs, 3H), 2.08 (m, 2H), 1.83 (m, 2H), 1.73 (m, 2H), 1.51/1.41 (s, 18H ), 1.2 (t, 6h); MSVR-ERI (m/z): [M+H] ⁺ , calculated for C ₃₅ H ₅₂ FN ₃ O ₁₀ PS: 756.3090; found: 756.3095.

Step B: Methyl 5-[3-[4-[3-[tert-butoxycarbonyl(methyl)amino]prop-1-ynyl]-2fluorophenoxy]propyl]-2-(3-diethoxyphosphorylpropylamino)thiazole-4-carboxylate.

Using the general HFIP deprotection procedure, 512 mg of product from step A (0.68 mmol, 1 eq.) gave 210 mg (47%) of the desired product.

1H NMR (500 MHz, DMSO-66) δ ppm 7.64 (brt, 1H), 7.31 (brd, 1H), 7.21 (dm, 1H), 7.13 (t, 1H), 4.23 (brs, 2H), 4.07 (t, 2H), 3.97 (m, 4H), 3.69 (s, 3H), 3.23 (m, 2H), 3.12 (t, 2H), 2.86 (brs, 3H), 2.00 (m, 2H), 1.84-1.63 (m, 4H), 1.41 (s, 3H), 1.21 (t, 6H); ¹³ C NMR (125 MHz, DMSO-06) δ ppm 129.1, 119.3, 115.4, 68.0, 61.4, 51.8, 44.7, 38.6, 33.8, 30.6, 28.5, 23.3, 16.7; MSVR-ERI (m/z): [M+H] ⁺ , calculated for C ₃₀ H ₄₄ FN ₃ O ₈ PS: 656.2565; found: 656.2555.

Step C: Methyl 5-[3-[4-[3-[tert-butoxycarbonyl(methyl)amino]prop-1-ynyl]-2fluorophenoxy]propyl]-2-[3-diethoxyphosphorylpropyl-[5-methyl-6- [^)-[3-(2-trimethylsilylethoxymethyl)1,3-benzothiazol-2-ylidene]amino]pyridazin-3-yl]amino]thiazol-4-carboxylate.

Using the general Buchwald reaction procedure II, starting with 450 mg of the product from step B (0.69 mmol, 1 eq.) and 363 mg of the product of synthesis 4a (0.89 mmol, 1.3 eq.) as the appropriate halide, 540 mg (77%) were obtained target product.

MSHR-ERI (m/z): [M+H] ⁺ , calculated for C ₄₈ H ₆₆ FN ₇ O ₉ PS ₂ Si: 1026.3849; found: 1026.3849.

Step D: 2-[[6-(1,3-benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]-(3-phosphonopropyl)amino]-5

- 257 046336

[3-[2-fluoro-4-[3-(methylamino)prop-1-ynyl]phenoxy]propyl]thiazole-4-carboxylic acid.

Using the general procedure of deprotection and hydrolysis of phosphonic acid derivatives followed by repurification by reverse phase preparative chromatography (C18, 0.1% TFA in water:MeCN), the TFA salt of the desired product was obtained from the product from step C.

MSHR-ERI (m/z): [M+H]+, calculated for C ₃₂ H ₃₄ FN ₇ O ₆ PS ₂ : 726.1728; found: 726.1707.

Example 238: 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(3carboxypropyl)amino)-5-(3-{4-[3-( dimethylamino)prop-1-in-1-yl]-2-fluorophenoxy}propyl)-1,3-thiazole4-carboxylic acid

HN

Step A: Methyl 2-[tert-butoxycarbonyl-(4-ethoxy-4-oxobutyl)amino]-5-[3-[4-[3(dimethylamino)prop-1-ynyl]-2-fluorophenoxy]propyl]thiazole -4-carboxylate.

Using the general alkylation procedure, 0.48 g (77%) of the target product was obtained from the synthesis product 1c and ethyl 4-bromobutanoate.

¹ H NMR (500 MHz, DMSO-de) δ ppm 7.28 (dd, 1H), 7.20 (dm, 1H), 7.12 (t, 1H), 4.10 (t, 2H), 4.04 (t, 2H), 4.00 (q, 2H), 3.75 (s, 3H), 3.41 (s, 2H), 3.22 (t, 2H), 2.31 (t, 2H), 2.22 (s, 6H), 2.08 (m, 2H), 1.91 (m, 2H), 1.50 (s, 9H), 1.13 ( t, 3H); ¹³ C NMR (125 MHz, DMSO-de) δ ppm 128.9, 119.2, 115.4, 68.1, 60.3, 52.1, 48.2, 46.0, 44.3, 31.2, 30.5, 28.0, 23.2, 23.2, 14.5; MSHR-ERI (m/z): [M+H] ⁺ , calculated for C30H41FN3O7S: 606.2649; found: 606.2640.

Step B: Methyl 5-[3-[4-[3-(dimethylamino)prop-1-ynyl]-2-fluorophenoxy]propyl]-2-[(4-ethoxy-4oxobutyl)amino]thiazole-4-carboxylate.

Using the general HFIP deprotection procedure, 0.32 g (83%) of the desired product was obtained from the product from step A.

¹ H NMR (400 MHz, DMSO^) δ ppm 7.61 (t, 1H), 7.30 (dd, 1H), 7.20 (dm, 1H), 7.12 (t, 1H), 4.07 (t, 2H), 4.04 (q, 2H), 3.69 (s, 3H), 3.42 (s, 2H), 3.12 (t, 2H), 3.118 (q, 2H), 2.35 (t, 2H), 2.23 (s, 6H), 2.00 (m, 2H), 1.76 (m, 2H), 1.17 ( t, 3H); ¹³ C NMR (100 MHz, DMSO^) δ ppm 173.1, 164.6, 163, 128.9, 119.2, 115.4, 68.0, 60.3, 51.7, 48.2, 44.3, 43.7, 31.3, 30.6, 24.5, 23.3, 14.6; MSHR-ERI (m/z): [M+H] ⁺ , calculated for C25H33FN3O5S: 506.2125; found: 506.2114.

Step C: methyl 5-[3-[4-[3-(dimethylamino)prop-1-ynyl]-2-fluorophenoxy]propyl]-2-[(4-ethoxy-4oxobutyl)-[5-methyl-6- [^)-[3-(2-trimethylsilylethoxymethyl)-1,3-benzothiazol-2ylidene]amino]pyridazin-3-yl]amino]thiazol-4-carboxylate.

Using the general procedure II of the Buchwald reaction, starting from the product from stage B and the compound Synthesis 4a, 0.26 g (48%) of the target product was obtained.

¹ H NMR (500 MHz, DMSO-de) δ ppm 7.84 (d, 1H), 7.73 (s, 1H), 7.47 (dd, 1H), 7.44 (td, 1H), 7.30 (dd, 1H), 7.25 (td, 1H), 7.21 (dd, 1H), 7.15 (t, 1H), 5.86 (s, 2H), 4.38 (br., 2H), 4.14 (t, 2H), 4.02 (q, 2H), 3.78 (s, 3H), 3.72 (t, 2H), 3.38 (s, 2H), 3.26 (t, 2H), 2.46 (s, 3H), 2.44 (m, 2H), 2.19 (s, 6H), 2.13 (m, 2H), 1.97 (m, 2H), 1.14 ( t, 3H), 0.92 (t, 2H), -0.11 (s, 9H); MSHR-ERI (m/z): [M+H] ⁺ , calculated for C42H55FN7O6S2Si: 876.3409; found: 876.3403; HRMS-ERI (m/z): [M+H] ⁺ , calculated for C42H ₅₃ FN ₇ O ₆ S ₂ Si (double Me-ester): 862.3252; found: 862.3237.

Step D: 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(3-carboxypropyl)amino)5-(3-{4-[3- (dimethylamino)prop-1-yn-1-yl]-2-fluorophenoxy}propyl)-1,3-thiazol-4-carboxylic acid.

Using the general deprotection and hydrolysis procedure followed by repurification by reverse phase preparative chromatography (C18, 0.1% TFA in water:MeCN), the TFA salt of the desired product was obtained from the product from step C.

MSVR-ERI (m/z): [M+H] ⁺ , calculated for C ₃₄ H ₃₅ FN ₇ O ₅ S ₂ : 704.2125; found: 704.2116.

Example 239: 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(3phosphonopropyl)amino)-5-(3-{4-[3-( dimethylamino)prop-1-in-1-yl]-2-fluorophenoxy}propyl)-1,3-thiazole4-carboxylic acid

- 258 046336

Step A: Methyl 2-[tert-butoxycarbonyl(3-diethoxyphosphorylpropyl)amino]-5-[3-[4-[3(dimethylamino)prop-1-ynyl]-2-fluorophenoxy]propyl]thiazole-4-carboxylate.

Using the general alkylation procedure, starting from the synthesis product 1c and 1-bromo-3diethoxyphosphorylpropane as a suitable halide, 0.52 g (77%) of the target product was obtained.

¹ H NMR (500 MHz, DMSO-dg) δ ppm 7.29 (dd, 1H), 7.20 (dm, 1H), 7.12 (t, 1H), 4.10 (t, 2H), 4.04 (t, 2H), 3.97 (m, 4H), 3.75 (s, 3H), 3.43 (s, 2H), 3.22 (t, 2H), 2.23 (s, 6H), 2.08 (m, 2H), 1.83 (m, 2H), 1.73 (m, 2H), 1.51 (s, 9H), 1.21 ( t, 6H); ¹³ C NMR (125 MHz, DMSOD ₆ ) δ ppm 156.5/153.0, 128.9, 119.2, 115.4, 68.1, 61.4, 52.1, 48.1, 46.9, 44.2, 30.5, 28.1, 23.2, 22.5, 21.3, 16.7; MSHR-ERI (m/z): [M+H]+, calculated for C ₃₁ H ₄₆ FN ₃ O ₈ PS: 670.2727; found: 670.2716.

Step B: Methyl 2-(3-diethoxyphosphorylpropylamino)-5-[3-[4-[3-(dimethylamino)prop-1-ynyl]-2fluorophenoxy]propyl]thiazole-4-carboxylate.

Using the general HFIP deprotection procedure, 0.37 g (86%) of the desired product was obtained from the product from step A.

¹ H NMR (500 MHz, DMSO^) δ ppm 7.64 (t, 1H), 7.30 (dd, 1H), 7.21 (dm, 1H), 7.12 (t, 1H), 4.07 (t, 2H), 3.99/3.95 (m+m, 4H), 3.69 (s, 3H), 3.43 (s, 2H), 3.23 (m, 2H), 3.12 (t, 2H), 2.23 (s, 6H), 2.00 (m, 2H), 1.84-1.64 (m, 4H), 1.21 (t , 6H); MSVR-ERI (m/z): [M+H]+, calculated for C ₂₆ H ₃₈ FN ₃ O ₆ PS: 570.2203; found: 570.2183.

Step C: methyl 2-[3-diethoxyphosphorylpropyl-[5-methyl-6-[(7)-[3-(2-trimethylsilylethoxymethyl)1,3-benzothiazol-2-ylidene]amino]pyridazin-3-yl]amino ]-5-[3-[4-[3-(dimethylamino)prop-1-ynyl]-2fluorophenoxy]propyl]thiazole-4-carboxylate.

Using the general Buchwald reaction procedure II, starting from the product from step B and the product of synthesis 4a as a suitable halide, 0.51 g (87%) of the desired product was obtained.

¹ H NMR (500 MHz, DMSO^) δ ppm 7.84 (d, 1H), 7.76 (s, 1H), 7.47 (dd, 1H), 7.44 (td, 1H), 7.30 (dd, 1H), 7.25 (td, 1H), 7.21 (dd, 1H), 7.15 (t, 1H), 5.86 (s, 2H), 4.40 (brt., 2H), 4.14 (t, 2H), 3.99 (m, 4H), 3.77 (s, 3H), 3.72 (t, 2H), 3.38 (br., 2H), 3.27 (t, 2H), 2.46 (s, 3H), 2.19 (s, 6H), 2.13 (m, 2H), 1.90 (m, 2H), 1.89 (m, 2H), 1.20 (t, 6H), 0.92 (t, 2H), -0.11 (s, 9H); ¹³ C NMR (125 MHz, DMSO^) δ ppm 163.1, 128.9, 127.2, 123.5, 123.2, 119.2, 117.4, 115.5, 112.0, 72.9, 68.5, 66.7, 61.5, 52.0, 48.1, 48.1, 44.2, 30.9, 23.1, 22.1, 20.5, 17.8, 17.8, 16.7, -1.0 ; MSHR-ERI (m/z): [M+H] ⁺ , calculated for C ₄₄ H ₆₀ FN ₇ O ₇ PS ₂ Si: 940.3487; found: 940.3471.

Step D: 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(3-phosphonopropyl)amino)5-(3 - {4-[3- (dimethylamino)prop-1-yn-1-yl]-2-fluorophenoxy}propyl)-1,3-thiazol-4-carboxylic acid.

Using the general procedure for deprotection and hydrolysis of phosphonic acid derivatives, followed by repurification by reverse phase preparative chromatography (C18, 0.1% TFA in water: MeSA), the TFA salt of the desired product was obtained from the product from step C.

MSVR-ERI (m/z): [M+H]+, calculated for C ₃₃ H ₃₆ FN ₇ O ₆ PS ₂ : 740.1890; found: 740.1871.

Example 240: 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(3-sulfopropyl)amino)5-(3 - {4-[3- (dimethylamino)prop-1 -yn-1 -yl]-2-fluorophenoxy}propyl)-1,3-thiazol-4-carboxylic acid

ABOUT

HN

Step A: 3-[tert-butoxycarbonyl-[5-[3-[4-[3-(dimethylamino)prop-1-ynyl]-2fluorophenoxy]propyl]-4-methoxycarbonylthiazol-2-yl]amino]propan-1 -sulfonic acid.

- 259 046336

Using the general alkylation procedure, starting from the synthesis product 1c and sodium 3-bromopropane-1sulfonate as a suitable halide, 0.50 g (77%) of the desired product was obtained.

¹ H NMR (500 MHz, DMSOC) δ ppm 7.33 (dd, 1H), 7.23 (dm, 1H), 7.15 (t, 1H), 4.10 (t, 2H), 4.04 (t, 2H), 3.76 (s, 3H), 3.62 (s, 2H), 3.23 (t, 2H), 2.37 (s, 6H), 2.36 (m, 2H), 2.09 (m, 2H), 1.92 (m, 2H), 1.51 (s, 9H); ¹³ C NMR (125 MHz, DMSOC) δ ppm 129.1, 119.3, 115.4, 68.1, 52.0, 49.6, 47.9, 46.1, 43.8, 30.4, 28.1, 24.7, 23.1; MSVR-ERI (m/z): [M+H]+, calculated for C ₂₇ H ₃₇ FN ₃ O ₈ S ₂ : 614.2006; found: 614.1993.

Step B: 3-[[5-[3-[4-[3-(dimethylamino)prop-1-ynyl]-2-fluorophenoxy]propyl]-4methoxycarbonylthiazol-2-yl]amino]propane-1-sulfonic acid.

Using the general HFIP deprotection procedure, 0.26 g (63%) of the desired product was obtained from the product from step A.

¹ H NMR (500 MHz, DMSOC) δ ppm 7.61 (t, 1H), 7.33 (dd, 1H), 7.24 (dd, 1H), 7.14 (t, 1H), 4.07 (t, 2H), 3.69 (s, 3H), 3.65 (s, 2H), 3.21 (q, 2H), 3.12 (t, 2H), 2.46 (t, 2H), 2.39 (s, 6H), 2.00 (qn, 2H), 1.80 (qn, 2H); ¹³ C NMR (125 MHz, DMSOC) δ ppm 164.7, 163.0, 151.5, 147.6, 136.9, 136.2, 129.1, 119.3, 115.5, 114.8, 85.2, 83.4, 68.1, 51.7, 49.6, 47.9, 44.1, 43.8, 30.6, 25.5, 23.3; HRMS-ERI (m/z): [M+Na] ⁺ , calculated for C ₂₂ H ₂ 8FN ₃ NaO6S ₂ : 536.1301; found: 536.1293.

Step C: 3-[[5-[3-[4-[3-(dimethylamino)prop-1-ynyl]-2-fluorophenoxy]propyl]-4methoxycarbonylthiazol-2-yl]-[5-methyl-6-[ ^)-[3-(2-trimethylsilylethoxymethyl)-1,3-benzothiazol-2ylidene]amino]pyridazin-3-yl]amino]propane-1-sulfonic acid.

Using the general Buchwald reaction procedure II, starting from the product from step B and the product of synthesis 4a as a suitable halide, 0.27 g (66%) of the desired product was obtained.

¹ H NMR (500 MHz, DMSOC) δ ppm 7.95 (s, 1H), 7.84 (d, 1H), 7.46 (d, 1H), 7.43 (t, 1H), 7.40 (dd, 1H), 7.28 (d, 1H), 7.25 (t, 1H), 7.19 (t, 1H), 5.86 (s, 2H), 4.48 (t, 2H), 4.16 (t, 2H), 3.90 (br., 2H), 3.78 (s, 3H), 3.72 (t, 2H), 3.27 (t, 2H), 2.57 (br., 6H), 2.53 (t, 2H), 2.45 (s, 3H), 2.14 (m, 2H), 2.00 (m, 2H), 0.92 (t, 2H), - 0.11 (s, 9H); ¹³ C NMR (125 MHz, DMSOC) δ ppm 163.2, 156.1, 134.9, 129.4, 127.2, 123.4, 123.2, 119.4, 117.8, 115.5, 111.9, 72.9, 68.5, 66.7, 52.0, 48.7, 47.3, 46.2, 42.9 , 30.9, 23.6, 23.1, 17.9, 17.8, -0.9 ; MSHR-ERI (m/z): [M+H] ⁺ , calculated for C ₄₀ H ₅₁ FN ₇ O ₇ S ₃ Si: 884.2765; found: 884.2757.

Step D: 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(3-sulfopropyl)amino)-5(3-{ 4-[3- (dimethylamino)prop-1-yn-1-yl]-2-fluorophenoxy}propyl)-1,3-thiazol-4-carboxylic acid.

Using the general deprotection and hydrolysis procedure followed by repurification by reverse phase preparative chromatography (C18, 0.1% TFA in water:MeCN), the TFA salt of the desired product was obtained from the product from step C.

MSHR-ERI (m/z): [M+H] ⁺ , calculated for C ₃₃ H ₃₅ FN ₇ O ₆ S ₃ : 740.1795; found: 740.1786.

Example 241: 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(4-sulfobutyl)amino)5-(3-{ 4-[3- (dimethylamino)prop-1-yn-1-yl]-2-fluorophenoxy}propyl)-1,3-thiazol-4-carboxylic acid

ABOUT

HN

Step A: 4-[tert-butoxycarbonyl-[5-[3-[4-[3-(dimethylamino)prop-1-ynyl]-2fluorophenoxy]propyl]-4-methoxycarbonylthiazol-2-yl]amino]butan-1 -sulfonic acid.

Using the general alkylation procedure, 0.29 g of the target product was obtained from oxathiane 2,2-dioxide and synthesis product 1c.

¹ H NMR (500 MHz, DMSOC) δ ppm 7.34 (dd, 1H), 7.24 (dd, 1H), 7.14 (t, 1H), 4.10 (t, 2H), 3.98 (t, 2H), 3.75 (s, 3H), 3.74 (s, 2H), 3.22 (t, 2H), 2.46 (s, 6H), 2.42 (t, 2H), 2.09 (qn, 2H), 1.65 (qn, 2H), 1.56 (qn, 2H), 1.51 (s, 9H); ¹³ C NMR (125 MHz, DMSOC) δ ppm 129.1, 119.3, 115.5, 68.2, 52.0, 51.7, 47.7, 46.4, 43.5, 30.4, 28.2, 27.4, 23.2, 22.9; HRMS-ERI (m/z): [M+Na] ⁺ , calculated for C ₂ 8H ₃ 8FN ₃ NaO8S ₂ : 650.1982; found: 650.1974.

Step B: 4-[[5-[3-[4-[3-(dimethylamino)prop-1-ynyl]-2-fluorophenoxy]propyl]-4methoxycarbonylthiazol-2-yl]amino]butane-1-sulfonic acid.

Using the general HFIP deprotection procedure, 0.12 g of the desired product was obtained from the product from step A.

MSHR-ERI (m/z): [M+H] ⁺ , calculated for C ₂₃ H ₃₁ FN ₃ O ₆ S ₂ : 528.1638; found: 528.1632.

Step C: 4-[[5-[3-[4-[3-(dimethylamino)prop-1-ynyl]-2-fluorophenoxy]propyl]-4methoxycarbonylthiazol-2-yl]-[5-methyl-6-[ ^)-[3-(2-trimethylsilylethoxymethyl)-1,3-benzothiazol-2

- 260 046336 ylidene]amino]pyridazin-3-yl]amino]butan-1-sulfonic acid.

Using the general procedure II for the Buchwald reaction, 0.16 g of the target product was obtained from the product from stage B and the product of synthesis 4a.

1H NMR (500 MHz, DMSO-dg) δ ppm 7.83 (d, 1H), 7.79 (s, 1H), 7.46 (d, 1H), 7.44 (t, 1H), 7.43 (dd, 1H), 7.31 (dm, 1H), 7.25 (td, 1H), 7.18 (t, 1H), 5.86 (s, 2H), 4.34 (t, 2H), 4.28 (s, 2H), 4.17 (t, 2H), 3.76 (s, 3H), 3.73 (t, 2H), 3.28 ( t, 2H), 2.86 (s, 6H), 2.51 (m, 2H), 2.46 (s, 3H), 2.15 (m, 2H), 1.73 (m, 2H), 1.67 (m, 2H), 0.92 (t , 2H), -0.11 (s, 9H); ¹³ C NMR (125 MHz, DMSO-06) δ ppm 129.6, 127.2, 123.4, 123.2, 119.6, 117.8, 115.4, 111.9, 72.9, 68.4, 66.8, 51.9, 51.3, 47.0, 47.0, 42.2, 30.7, 26.2, 23.1, 2 2.9, 17.8, 17.8, -0.9; HRMS-ERI (m/z): [M+H]+, calculated for C ₄₁ H ₅₃ FN ₇ O ₇ S ₃ Si: 898.2922; found: 898.2907.

Step D: 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(4-sulfobutyl)amino)-5(3-{4-[3 - (dimethylamino)prop-1-yn-1-yl]-2-fluorophenoxy}propyl)-1,3-thiazol-4-carboxylic acid.

Using the general deprotection and hydrolysis procedure, 64 mg of the desired product was obtained from the product from step C.

MSVR-ERI (m/z): [M+H] ⁺ , calculated for C ₃₄ H ₃₇ FN ₇ O ₆ S ₃ : 754.1951; found: 754.1930.

Example 242: 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(4-phosphonobutyl)amino)5-(3-{4-[3 - (dimethylamino)prop-1-yn-1-yl]-2-fluorophenoxy}propyl)-1,3-thiazol-4-carboxylic acid

ABOUT

HN

Step A: Methyl 2-[tert-butoxycarbonyl(4-diethoxyphosphorylbutyl)amino]-5-[3-[4-[3(dimethylamino)prop-1-ynyl]-2-fluorophenoxy]propyl]thiazole-4-carboxylate.

Using the general alkylation procedure, starting from 1-bromo-4-diethoxyphosphorylbutane and synthesis product 1c, 1.28 g of the target product was obtained.

¹ H NMR (500 MHz, DMSO-dg) δ ppm 7.29 (dd, 1H), 7.20 (dm, 1H), 7.13 (t, 1H), 4.10 (t, 2H), 4.01 (t, 2H), 3.93 (m, 4H), 3.75 (s, 3H), 3.42 (s, 2H), 3.22 (t, 2H), 2.23 (s, 6H), 2.08 (m, 2H), 1.78 (m, 2H), 1.71 (m, 2H), 1.51 (s, 9H), 1.45 ( m, 2H), 1.18 (t, 6H); ¹³ C NMR (125 MHz, DMSO-06) δ ppm 128.9, 119.2, 115.4, 68.1, 61.2, 52.0, 48.1, 45.9, 44.2, 30.5, 28.8, 28.0, 24.5, 23.2, 19.9, 16.7; MSVR-ERI (m/z): [M+H] ⁺ , calculated for C ₃₂ H ₄₈ FN ₃ O ₈ PS: 684.2884; found: 684.2859.

Step B: Methyl 2-(4-diethoxyphosphorylbutylamino)-5-[3-[4-[3-(dimethylamino)prop-1-ynyl]-2fluorophenoxy]propyl]thiazole-4-carboxylate.

Using the general HFIP deprotection procedure, 0.92 g of the desired product was obtained from the product from step A.

1H NMR (500 MHz, DMSO^b) δ ppm 7.61 (t, 1H), 7.32 (dd, 1H), 7.22 (dm, 1H), 7.13 (t, 1H), 4.07 (t, 2H), 3.95 (m, 4H), 3.69 (s, 3H), 3.52 (brs., 2H), 3.17 (q, 2H), 3.12 (t, 2H), 2.30 (s, 6H), 2.00 (m, 2H), 1.73 (m, 2H), 1.58 (m, 2H), 1.50 (m, 2H), 1.21 (t, 6H); ¹³ C NMR (125 MHz, DMSO-06) δ ppm 163.0, 129.0, 119.2, 115.4, 68.0, 61.2, 51.7, 48.0, 44.0, 43.8, 30.6, 29.8, 24.6, 23.3, 20.1, 16.8; MSVR-ERI (m/z): [M+H] ⁺ , calculated for C ₂₇ H ₄₀ FN ₃ O ₆ PS: 584.2359; found: 584.2340.

Step C: methyl 2-[4-diethoxyphosphorylbutyl-[5-methyl-6-[(7)-[3-(2-trimethylsilylethoxymethyl)1,3-benzothiazol-2-ylidene]amino]pyridazin-3-yl]amino ]-5-[3-[4-[3-(dimethylamino)prop-1-ynyl]-2fluorophenoxy]propyl]thiazole-4-carboxylate.

Using the general procedure II for the Buchwald reaction, starting from the product from stage B and the product of synthesis 4a, 0.69 g of the target product was obtained.

1H NMR (500 MHz, DMSO-06) δ ppm 7.84 (dm, 1H), 7.72 (s, 1H), 7.47 (dm, 1H), 7.44 (m, 1H), 7.31 (dd, 1H), 7.26 (m, 1H), 7.21 (dm, 1H), 7.16 (t, 1H), 5.86 (s, 2H), 4.41 (t, 2H), 4.15 (t, 2H), 3.94/3.91 (m+m, 4H), 3.78 (s, 3H), 3.72 (m, 2H ), 3.38 (s, 2H), 3.27 (t, 2H), 2.46 (s, 3H), 2.20 (s, 6H), 2.12 (m, 2H), 1.86 (m, 2H), 1.78 (m, 2H) , 1.54 (m, 2H), 1.18 (t, 6H), 0.91 (m, 2H), -0.11 (s, 9H); MSHR-ERI (m/z): [M+H] ⁺ , calculated for C ₄₅ H ₆₂ FN ₇ O ₇ PS ₂ Si: 954.3643; found: 954.3627.

Step D: 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(4-phosphonobutyl)amino)-5(3-{4-[3 - (dimethylamino)prop-1-yn-1-yl]-2-fluorophenoxy}propyl)-1,3-thiazol-4-carboxylic acid.

Using the general procedure for deprotection and hydrolysis of phosphonic acid derivatives, 288 mg of the desired product was obtained from the product from step C.

MSVR-ERI (m/z): [M+H] ⁺ , calculated for C ₃₄ H ₃₈ FN ₇ O6PS ₂ : 754.2047; found: 754.2036.

Example 243: 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl} (4

- 261 046336 carboxybutyl)amino)-5-(3-{4-[3-(dimethylamino)prop-1-in-1-yl]-2-fluorophenoxy}propyl)-1,3-thiazol-4carboxylic acid

Step A: Methyl 2-[tert-butoxycarbonyl-(5-methoxy-5-oxopentyl)amino]-5-[3-[4-[3(dimethylamino)prop-1-ynyl]-2-fluorophenoxy]propyl]thiazole -4-carboxylate.

Using the general alkylation procedure, starting from methyl 5-bromopentanoate and synthesis product 1c, 0.99 g of the target product was obtained.

1H NMR (500 MHz, DMSO-06) δ ppm 7.28 (dd, 1H), 7.20 (dm, 1H), 7.13 (t, 1H), 4.10 (t, 2H), 4.00 (t, 2H), 3.75 (s, 3H), 3.56 (s, 3H), 3.41 (s, 2H), 3.22 (t, 2H), 2.37 (t, 2H), 2.22 (s, 6H), 2.08 (m, 2H), 1.63 (m, 2H), 1.51 (m, 2H), 1.51 ( s, 9H); ¹³ C NMR (125 MHz, DMSO-06) δ ppm 128.9, 119.2, 115.4, 68.1, 52.1, 51.7, 48.2, 46.0, 44.3, 33.2, 30.5, 28.1, 27.3, 23.1, 22.0; MSHR-ERI (m/z): [M+H]+, calculated for C30H41FN3O7S: 606.2649; found: 606.2644.

Step B: Methyl 5-[3-[4-[3-(dimethylamino)prop-1-ynyl]-2-fluorophenoxy]propyl]-2-[(5-methoxy-5oxopentyl)amino]thiazole-4-carboxylate.

Using the general HFIP deprotection procedure, 0.72 g of the desired product was obtained from the product from step A.

¹ H NMR (500 MHz, DMSO-06) δ ppm 7.59 (t, 1H), 7.29 (dd, 1H), 7.20 (dm, 1H), 7.12 (t, 1H), 4.07 (t, 2H), 3.69 (s, 3H), 3.58 (s, 3H), 3.41 (s, 2H), 3.16 (q, 2H), 3.12 (t, 2H), 2.33 (t, 2H), 2.22 (s, 6H), 2.00 (m, 2H), 1.56 (m, 2H), 1.51 ( m, 2H); ¹³ C NMR (125 MHz, DMSO-06) δ ppm 173.7, 164.6, 163.0, 128.9, 119.2, 115.4, 85.1, 84.1, 68.0, 51.7, 51.7, 48.2, 44.3, 44.0, 33.4, 30.6, 28.5, 23.3, 22.4; MSVR-ERI (m/z): [M+H] ⁺ , calculated for C ₂₅ H ₃₃ FN ₃ O ₅ S: 506.2125; found: 506.2123.

Step C: methyl 5-[3-[4-[3-(dimethylamino)prop-1-ynyl]-2-fluorophenoxy]propyl]-2-[(5-methoxy-5oxopentyl)-[5-methyl-6- [(2)-[3-(2-trimethylsilylethoxymethyl)-1,3-benzothiazol-2ylidene]amino]pyridazin-3-yl]amino]thiazol-4-carboxylate.

Using the general procedure II for the Buchwald reaction, starting from the product from stage B and the product of synthesis 4a, 0.79 g of the target product was obtained.

1H NMR (500 MHz, DMSO-06) δ ppm 7.83 (dm, 1H), 7.65 (s, 1H), 7.45 (dm, 1H), 7.43 (m, 1H), 7.30 (dd, 1H), 7.25 (m, 1H), 7.20 (dm, 1H), 7.15 (t, 1H), 5.85 (s, 2H), 4.37 (t, 2H), 4.14 (t, 2H), 3.77 (s, 3H), 3.71 (m, 2H), 3.56 (s, 3H), 3.39 ( s, 2H), 3.26 (t, 2H), 2.45 (s, 3H), 2.43 (t, 2H), 2.20 (s, 6H), 2.12 (m, 2H), 1.71 (m, 2H), 1.60 (m , 2Н), 0.91 (m, 2Н), -0.11 (s, 9h); MSHR-ERI (m/z): [M+H] ⁺ , calculated for C43H55FN ₇ O6S ₂ Si: 876.3409; found: 876.3378.

Step D: 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(4-carboxybutyl)amino)-5(3-{4-[3- (dimethylamino)prop-1-yn-1-yl]-2-fluorophenoxy}propyl)-1,3-thiazol-4-carboxylic acid.

Using the general deprotection and hydrolysis procedure, 296 mg of the desired product was obtained from the product from step C.

MSHR-ERI (m/z): [M+H] ⁺ , calculated for C ₃₅ H ₃₇ FN ₇ O ₅ S ₂ : 718.2282; found: 718.2263.

Example 244: 2-[(3-aminopropyl){6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}amino]5-(3-{2-fluoro-4 -[3-(methylamino)prop-1-yn-1-yl]phenoxy} propyl)-1,3-thiazol-4-carboxylic acid

Example 245: 2-[(3-azidopropyl){6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}amino]- 262 046336

5-(3-{2-fluoro-4-[3-(methylamino)prop-1-yn-1-yl]phenoxy}propyl)-1,3-thiazole-4-carboxylic acid

Example 246: 2-[{6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino]-5-[3-(4-{3[( dimethylamino)methyl]bicyclo[1.1.1]pentan-1-yl}-2-fluorophenoxy)propyl]-1,3-thiazol-4-carboxylic acid

Example 247: 6-[{6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl} (methyl)amino]-3-(1-{[3(2-{ [(3S)-3,4-dihydroxybutyl]amino}ethoxy)-5,7-dimethyladamantan-1-yl]methyl}-5-methyl-1H-pyrazol4-yl)pyridine-2-carboxylic acid

Using the general procedure for amine substitution and hydrolysis starting from the synthesis product 14 and 2-[(4S)-2,2-dimethyl-1,3-dioxolan-4-yl]ethanamine as a suitable amine, the desired product was obtained.

MSHR-ERI (m/z): [M+H]+, calculated for C ₄₂ H ₅₄ N9O ₅ S: 796.3969; found: 796.3965.

Example 248: 6-[{6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino]-3-(1-{[3(2-{ [(3R)-3,4-dihydroxybutyl]amino}ethoxy)-5,7-dimethyladamantan-1-yl]methyl}-5-methyl-1H-pyrazol4-yl)pyridine-2-carboxylic acid

- 263 046336

Example 249: 6-[{6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino]-3-[1-({3,5dimethyl-7 -[2-(methylamino)ethoxy] adamantan-1 -yl } methyl)-5-methyl-1 H-pyrazol-4-yl] pyridin-2-carboxylic acid

Example 250: 6-[{6-[(1,3-Benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino]-3-[1-({3[2-( dimethylamino)ethoxy]-5,7-dimethyladamantan-1-yl}methyl)-5-methyl-1H-pyrazol-4-yl]pyridine-2carboxylic acid

Using the general procedure for amine substitution and hydrolysis starting from synthesis 14 and dimethylamine as a suitable amine, the desired product was obtained.

MSHR-ERI (m/z): [M+H]+, calculated for C ₄₀ H ₅₀ N ₉ O ₃ S: 736.3757; found: 736.3751.

Example 251: 6-[{6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino]-3-{1-[(3,5dimethyl-7 -{2-[(2-sulfoethyl)amino]ethoxy}adamantan-1-yl)methyl]-5-methyl-1H-pyrazol-4-yl}pyridine2-carboxylic acid

- 264 046336

Example 252: 6-[{6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino]-3-{1-[(3,5dimethyl-7 - {2-[methyl(2-sulfoethyl)amino]ethoxy} adamantan-1 -yl)methyl]-5-methyl-1 H-pyrazol-4yl}pyridine-2-carboxylic acid

Example 253: 2-[{6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(pent-4-yn-1-yl)amino]5-(3- {2-fluoro-4-[3-(methylamino)prop-1-yn-1-yl]phenoxy}propyl)-1,3-thiazol-4-carboxylic acid

Step A: methyl 2-[tert-butoxycarbonyl(5-trimethylsilylpent-4-ynyl)amino]-5-[3-(2-fluoro-4iodophenoxy)propyl]-1,3-thiazol-4-carboxylate.

Using the general Mitsunobu reaction procedure, starting with the synthesis product 1a (10 mmol) as a suitable carbamate and 5-trimethylsilylpent-4-yn-1-ol (2 eq.) as a suitable alcohol, 6.0 g (89%) of the target product was obtained product.

¹ H NMR (500 MHz, DMSO-de) δ ppm 7.91 (dm, 1H), 7.69 (s, 1H), 7.53 (dm, 1H), 7.39 (m, 1H), 7.34 (dd, 1H), 7.25 (dm, 1H), 7.21 (m, 1H), 7.19 (t, 1H), 4.38 (t, 2H), 4.16 (t, 2H), 3.87 (s, 2H), 3.27 (t, 2H), 2.88 (t, 1H),

- 265 046336

2.51 (s, 3H), 2.46 (s, 3H), 2.31 (m, 2H), 2.14 (m, 2H), 1.91 (m, 2H); MSHR-ERI (m/z): [M+H]+, calculated for C ₃₄ H ₃₃ FN ₇ O ₃ S ₂ : 670.2070; found: 670.2052.

Step B: Methyl 5-[3-(2-fluoro-4-iodophenoxy)propyl]-2-(5-trimethylsilylpent-4-ynylamino)-1,3thiazole-4-carboxylate.

Using the general HFIP deprotection procedure, starting with the product from Step A (6.0 g, 8.9 mmol, 1 eq.) as the appropriate Boc-protected amine, 4.46 g (87%) of the title product was obtained.

¹ H NMR (500 MHz, DMSO-de) δ ppm 7.61 (t, 1H), 7.59 (dd, 1H), 7.45 (dm, 1H), 6.97 (t, 1H), 4.03 (t, 2H), 3.69 (s, 3H), 3.22 (m, 2H), 3.11 (t, 2H), 2.28 (t, 2H), 1.99 (m, 2H), 1.68 (m, 2H), 0.11 (s, 9H); ¹³ C NMR (125 MHz, DMSOC) δ ppm 134.0, 124.9, 117.6, 107.8, 85.1, 68.1, 51.7, 43.5, 30.6, 28.0, 23.3, 17.1, 0.6; MSVRERI (m/z): [M+H]+, calculated for C ₂₂ H ₂₉ FIN ₂ O ₃ SSi: 575.0697; found: 575.0695.

Step C: methyl 5-[3-[4-[3-[tert-butoxycarbonyl(methyl)amino]prop-1-ynyl]-2fluorophenoxy]propyl]-2-(5-trimethylsilylpent-4-ynylamino)-1, 3-thiazole-4-carboxylate.

Using the general Sonogashira reaction procedure, starting with the product from step B (4.46 g, 7.76 mmol, 1 eq.) and tert-butyl N-methyl-N-prop-2-ynylcarbamate (2.62 g, 15.53 mmol, 2 eq.) as suitable acetylene, 4.44 g (93%) of the target product were obtained.

¹ H NMR (500 MHz, DMSO-de) δ ppm 7.61 (t, 1H), 7.30 (brd, 1H), 7.21 (dm, 1H), 7.12 (t, 1H), 4.23 (brs, 2H), 4.07 (t, 2H), 3.69 (s, 3H), 3.22 (m, 2H), 3.12 (t, 2H), 2.87 (brs, 3H), 2.28 (t, 2H), 2.00 (m, 2H), 1.68 (m, 2H), 1.41 (s, 9H), 0.11 ( s, 9H); MSHR-ERI (m/z): [M+H]+, calculated for C ₃₁ H ₄₃ FN ₃ O ₅ SSi: 616.2677; found: 616.2659.

Step D: methyl 5-[3-[4-[3-[tert-butoxycarbonyl(methyl)amino]prop-1-ynyl]-2fluorophenoxy]propyl]-2-[[5-methyl-6-[^)- [3-(2-trimethylsilylethoxymethyl)-1,3-benzothiazol-2ylidene]amino]pyridazin-3-yl]-(5-trimethylsilylpent-4-ynyl)amino]-1,3-thiazol-4-carboxylate.

Using the general Buchwald reaction procedure I, starting with the product from step C (4.44 g, 7.2 mmol) as starting material and reacting in 1,4-dioxane at 120°C, 3.85 g (54%) of the desired product was obtained.

1H NMR (500 MHz, DMSO-de) δ ppm 7.83 (d, 1H), 7.64 (s, 1H), 7.45 (dd, 1H), 7.42 (td, 1H), 7.31 (brd., 1H), 7.24 (td, 1H), 7.21 (d, 1H), 7.15 (t, 1H), 5.85 (s, 2H), 4.37 (t, 2H), 4.20 (br., 2H), 4.14 (t, 2H), 3.77 (s, 3H), 3.71 (t, 2H), 3.25 (t, 2H), 2.84 (br., 3H), 2.44 (s, 3H), 2.37 (t, 2H), 2.12 (m, 2H), 1.91 (m, 2H), 1.4 (s, 9H), 0.91 (t, 2H), 0.09 (s, 9H), -0.12 (s, 9H); ¹³ C NMR (125 MHz, DMSO-de) δ ppm 163.1, 157.5, 155.2, 150.9, 137.6, 129.1, 127.2, 125.4, 123.4, 123.2, 119.3, 117.4, 115.4, 111.9, 107.5, 85.2, 72.9, 68.4, 66.7, 52.0, 46.5, 38.6, 33.8, 31.0, 28.5, 26.2, 23.2, 17.9, 17.8, 17.1, 0.5, -1.0; MSHR-ERI (m/z): [M+H]+, calculated for C49H65FN7O6S2Si2: 986.3960; found: 986.3932.

Step E: 2-[[6-(1,3-benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]-pent-4-ynylamino]-5-[3-[2fluoro-4-[3- (methylamino)prop-1-ynyl]phenoxy]propyl]-1,3-thiazole-4-carboxylic acid.

Using the general deprotection and hydrolysis procedure from the product from step D followed by purification by reverse phase preparative chromatography (C18, 25 mM NH4HCO3 in water:MeCN) the desired product was obtained.

MSHR-ERI (m/z): [M+H]+, calculated for C ₃₄ H ₃₃ FN ₇ O ₃ S ₂ : 670.2070; found: 670.2052.

Example 254: 2-[(4-Aminobutyl){6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}amino]-5(3 -{4-[3 - (dimethylamino)prop-1-yn-1-yl]-2-fluorophenoxy}propyl)-1,3-thiazol-4-carboxylic acid

Example 255: 2-[(4-Azidobutyl){6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}amino]-5(3 -{4-[3 - (dimethylamino)prop-1-yn-1-yl]-2-fluorophenoxy}propyl)-1,3-thiazol-4-carboxylic acid

- 266 046336

Example 256: 2-[{6-[(1,3-Benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl} (hex-5-yn-1 -yl)amino]-5(3- {4-[3-(dimethylamino)prop-1-yn-1-yl]-2-fluorophenoxy}propyl)-1,3-thiazol-4-carboxylic acid

Example 257: 2-[{6-[(1,3-Benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(pent-4-yn-1 -yl)amino]5 -(3 - {4-[3-(dimethylamino)prop-1-yn-1-yl]-2-fluorophenoxy}propyl)-1,3-thiazol-4-carboxylic acid

Example 258: 2-[(3-Azidopropyl){6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}amino]5 -(3 -{4-[3- (dimethylamino)prop-1 -yn-1 -yl]-2-fluorophenoxy}propyl)-1,3-thiazol-4-carboxylic acid

Example 259: 2-[(3-Aminopropyl){6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}amino]5 -(3 -{4-[3- (dimethylamino)prop-1 -yn-1 -yl]-2-fluorophenoxy}propyl)-1,3-thiazol-4-carboxylic acid

- 267 046336

Example 260: 2-[(4-Aminobutyl){6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}amino]-5(3-{2-fluoro-4 -[3-(methylamino)prop-1-yn-1-yl]phenoxy}propyl)-1,3-thiazol-4-carboxylic acid o.

Example 261: 2-[(4-Azidobutyl){6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}amino]-5(3-{2-fluoro-4 -[3-(methylamino)prop-1-yn-1-yl]phenoxy}propyl)-1,3-thiazol-4-carboxylic acid o.

Example 262: 2-[{6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(hex-5-yn-1-yl)amino]-5(3- {2-fluoro-4-[3-(methylamino)prop-1-yn-1-yl]phenoxy}propyl)-1,3-thiazol-4-carboxylic acid

Example 263: 2-[(4-Aminobutyl){6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}amino]-5(3-{2-fluoro-4 -[3-(methylamino)propyl]phenoxy}propyl)-1,3-thiazole-4-carboxylic acid

- 268 046336

Example 264: 2-[(4-Azidobutyl){6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}amino]-5(3-{2-fluoro-4 -[3-(methylamino)propyl]phenoxy}propyl)-1,3-thiazole-4-carboxylic acid

Example 265: 2-[{6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(hex-5-yn-1-yl)amino]-5(3- {2-fluoro-4-[3-(methylamino)propyl]phenoxy}propyl)-1,3-thiazole-4-carboxylic acid

Example 266: 5-[3-[4-[3-tert-Butoxycarbonyl(methyl)amino]prop-1-ynyl]-2-fluorophenoxy]propyl]2-[[5 -methyl-6-[(2)- [3-(2-trimethylsilylethoxymethyl)-1,3-benzothiazol-2-ylidene]amino]pyridazin-3-yl]pent-4-ynylamino]thiazol-4-carboxylic acid

A mixture of 500 mg of the product from Example 253, Step D, and 106 mg (2.5 mmol, 5 eq.) LiOHxH ₂ O in a mixture of 5 ml THF and 2 ml water was stirred at 40°C for 16 hours, and then purified by crystallization from Et ₂ O to obtain 356 mg (78%) of the target product.

MSHR-ERI (m/z): [M+H]+, calculated for C ₄₅ H ₅₅ FN ₇ O ₆ S ₂ Si: 900.3409; found: 900.3383

Example 267: 2-[5-Azidopentyl-[6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl]amino]-5[3-[2-fluoro-4- [3-(methylamino)prop-1-ynyl]phenoxy]propyl]-1,3-thiazol-4-carboxylic acid

- 269 046336

Step A: methyl 2-[tert-butoxycarbonyl-[5-[tert-butyl(dimethyl)silyl]oxypentyl]amino]-5-[3[4-[3-[tert-butoxycarbonyl(methyl)amino]prop-1 -inyl]-2-fluorophenoxy]propyl]-1,3-thiazol-4carboxylate.

Using the general Mitsunobu reaction procedure, starting with synthesis 1b (0.7 mmol) as the appropriate carbamate and 5-[tert-butyl(dimethyl)silyl]oxypentan-1-ol (1.5 eq.) as the appropriate alcohol, 494 mg was obtained (91%) of the target product.

LC/MS (C ₃ 9H60FN ₃ O8SSiNa) 800 [M+Na]+.

Step B: methyl 5-[3-[4-[3-[tert-butoxycarbonyl(methyl)amino]prop-1-ynyl]-2fluorophenoxy]propyl]-2-[5-[tert-butyl(dimethyl)silyl] hydroxypentylamino]-1,3-thiazole-4-carboxylate.

Using the general HFIP deprotection procedure, starting with the product from Step A (490 mg, 0.63 mmol, 1 eq.) as the appropriate Boc-protected amine, 240 mg (56%) of the desired product was obtained.

¹ H NMR (500 MHz, DMSO^) δ ppm 7.57 (br., 1H), 7.31 (d, 1H), 7.21 (d, 1H), 7.12 (t, 1H), 4.23 (br., 2H), 4.07 (t, 2H), 3.69 (s, 3H) , 3.56 (t, 2H), 3.14 (m, 2H), 3.11 (t, 2H), 2.86 (br., 3H), 2.00 (quint., 2H), 1.51 (m, 2H), 1.45 (m, 2H ), 1.41 (s, 9H), 1.33 (m, 2H), 0.84 (s, 9H), 0 (s, 6H); ¹³ C NMR (125 MHz, DMSO-06) δ ppm 129.1, 119.3, 115.4, 68.0, 62.8, 51.7, 44.5, 38.6, 33.8, 32.4, 30.6, 28.9, 28.5, 26.3, 23.3, 23.2, -4.9; MSVR-ERI (m/z): [M+H]+, calculated for C ₃₄ FN ₃ O ₆ SSi: 678.3408; found: 678.3393.

Step C: methyl 5-[3-[4-[3-[tert-butoxycarbonyl(methyl)amino]prop-1-ynyl]-2fluorophenoxy]propyl]-2-[5-[tert-butyl(dimethyl)silyl] oxypentyl-[5-methyl-6-[(Z)-[3-(2trimethylsilylethoxymethyl)-1,3-benzothiazol-2-ylidene]amino]pyridazin-3-yl]amino]-1,3-thiazol-4carboxylate.

Using the general Buchwald reaction procedure I, starting with the product from step B (235 mg) as starting material and reacting in 1,4-dioxane at 120° C., 220 mg (60%) of the desired product was obtained.

1H NMR (500 MHz, DMSO-06) δ ppm 7.81 (dd, 1H), 7.62 (s, 1H), 7.45 (dd, 1H), 7.42 (m, 1H), 7.28 (dd, 1H), 7.24 (m, 1H), 7.19 (m, 1H), 7.15 (t, 1H), 5.84 (s, 2H), 4.38 (t, 2H), 4.20 (s, 2H), 4.16 (t, 2H), 3.78 (s, 3H), 3.73 (t, 2H), 3.55 ( t, 2H), 3.26 (t, 2H), 2.85 (s, 3H), 2.44 (s, 3H), 2.13 (m, 2H), 1.71 (m, 2H), 1.51 (m, 2H), 1.41 (s , 9H), 1.40 (m, 2H), 0.92 (t, 2H), 0.80 (s, 9H), -0.04 (s, 6H), -0.10 (s, 9H); ¹³ C NMR (125 MHz, DMSO^) δ ppm 163.2, 147.5, 137.6, 129, 127.1, 123.5, 123.1, 119.3, 117.7, 115.7, 111.9, 73.0, 68.6, 66.8, 62.7, 51.8, 47.0, 38.6, 33.8, 32.4, 31.0, 28.5, 27.1, 26.2, 23.1, 23.0, 17.9, 17.8, -1.0, -5.0; MSHR-ERI (m/z): [M+H]+, calculated for C ₅₂ H ₇₅ FN ₇ O ₇ S ₂ Si ₂ : 1048.4692; found: 1048.4692.

Step D: Methyl 5-[3-[4-[3-[tert-butoxycarbonyl(methyl)amino]prop-1-ynyl]-2fluorophenoxy]propyl]-2-[5-hydroxypentyl-[5-methyl-6- [^)-[3-(2-trimethylsilylethoxymethyl)-1,3benzothiazol-2-ylidene]amino]pyridazin-3-yl]amino]-1,3-thiazol-4-carboxylate.

A mixture of the product from step C (0.3 mmol) and 18 mg camphorsulfonic acid (0.07 mmol, 0.25 eq.) in 2 ml of a 1:1 mixture of DCM and MeOH was stirred at 60°C for 1.5 h. The product was purified using flash column chromatography using heptane and EtOAc as eluents to obtain 195 mg (66%) of the desired product.

1H NMR (500 MHz, DMSO^) δ ppm 7.84 (d, 1H), 7.66 (s, 1H), 7.46 (d, 1H), 7.43 (t, 1H), 7.32 (d, 1H), 7.25 (t, 1H), 7.22 (d, 1H), 7.16 (t, 1H), 5.86 (s, 2H), 4.36 (t, 2H), 4.20 (s, 2H), 4.15 (t, 2H), 3.78 (s, 3H), 3.72 (t, 2H), 3.40 ( t, 2H), 3.26 (t, 2H), 2.84 (s, 3H), 2.46 (s, 3H), 2.12 (qn, 2H), 1.69 (qn, 2H), 1.49 (m, 2H), 1.40 (s , 9H), 1.4 (qn, 2H), 0.92 (t, 2H), -0.11 (s, 9H); ¹³ C NMR (125 MHz, DMSO^) δ ppm 129.1, 127.2, 123.4, 123.2, 119.3, 117.6, 115.5, 112.0, 72.9, 68.4, 66.7, 61.0, 52.0, 47.0, 38.5, 33.9, 32.5, 31.0, 28.5, 2 7.1, 23.2, 23.1, 17.9, 17.8, -1.0 ; HRMS-ERI (m/z): [M+Na]+, calculated for C ₄₆ H ₆₀ FN ₇ NaO ₇ S ₂ SiNa: 956.3641; found: 956.3646.

Step E: methyl 2-[5-azidopentyl-[5-methyl-6-[^)-[3-(2-trimethylsilylethoxymethyl)-1,3benzothiazol-2-ylidene]amino]pyridazin-3-yl]amino]- 5-[3-[4-[3-[tert-butoxycarbonyl(methyl)amino]prop-1-ynyl]-2-fluorophenoxy]propyl]-1,3-thiazol-4-carboxylate.

A mixture of 158 mg of the product from step D, 70 μl of triethylamine (0.50 mmol, 3 eq.) and 83 mg of ptolylsulfonyl 4-methylbenzenesulfonate (0.25 mmol, 1.5 eq.) in 1 ml of DCM was stirred for 5 h. After concentration, the residue was dissolved in 1 ml of acetonitrile and 133 mg of sodium azide (2.0 mmol, 12 eq.) was added and the resulting mixture was stirred at 80°C for 0.5 h. Product

- 270 046336 was purified by flash column chromatography using heptane and EtOAc as eluents to obtain 120 mg (73%) of the desired product.

1H NMR (500 MHz, DMSO^) δ ppm 7.84 (dm, 1H), 7.69 (s, 1H), 7.47 (dm, 1H), 7.44 (m, 1H), 7.32 (brd, 1H), 7.26 (m, 1H), 7.22 (dm, 1H), 7.17 (t, 1H), 5.86 (s, 2H), 4.37 (t, 2H), 4.20 (brs, 2H), 4.15 (t, 2H), 3.78 (s, 3H), 3.72 (m, 2H), 3.36 ( t, 2H), 3.27 (t, 2H), 2.84 (brs, 3H), 2.46 (s, 3H), 2.13 (m, 2H), 1.72 (m, 2H), 1.64 (m, 2H), 1.43 (m , 2Н), 1.40 (s, 9Н), 0.92 (m, 2Н), -0.11 (s, 9 Н); MSVR-ERI (m/z): [M+H] ⁺ , calculated for C46H60FN _w O6S2Si: 959.3892; found: 959.3892.

Step F: 2-[5-azidopentyl-[6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl]amino]-5-[3[2-fluoro-4- [3-(methylamino)prop-1-ynyl]phenoxy]propyl]-1,3-thiazole-4-carboxylic acid.

Using the general deprotection and hydrolysis procedure from the product from step E followed by purification by reverse phase preparative chromatography (C18, 0.1% TFA in water:MeCN) the desired product was obtained.

MSHR-ERI (m/z): [M+H] ⁺ , calculated for C ₃₄ H ₃₆ FN ₁₀ O ₃ S ₂ : 715.2397; found: 715.2391.

Example 268: 6-[[6-(1,3-Benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]methylamino]-3-[1-[[3-[2-(3hydroxypropylamino)ethoxy]- 5,7-dimethyl-1-adamantyl]methyl]-5-methylpyrazol-4-yl]pyridine-2carboxylic acid

Using the general procedure for amine substitution reaction and hydrolysis, starting from the synthesis product 14 and 3-aminopropan-1-ol as a suitable amine, the desired product was obtained.

MSHR-ERI (m/z): [M+H]+, calculated for C ₄₁ H52N9O ₄ S: 766.3863; found: 766.3860.

Example 269: 6-[[6-(1,3-Benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]methylamino]-3-[1-[[3-[2-(3methoxypropylamino)ethoxy]- 5,7-dimethyl-1-adamantyl]methyl]-5-methylpyrazol-4-yl]pyridine-2carboxylic acid

Using the general procedure of amine substitution reaction and hydrolysis starting from the synthesis product 14 and 3-methoxypropan-1-amine as a suitable amine, the desired product was obtained.

MSHR-ERI (m/z): [M+H] ⁺ , calculated for C ₄₂ H ₅₄ N ₉ O ₄ S: 780.4019; found: 780.4019.

Example 270: 6-[[6-(1,3-benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]methylamino]-3-[1-[[3,5dimethyl-7-(2-morpholinoethoxy) -1-adamantyl]methyl]-5-methylpyrazol-4-yl]pyridine-2-carboxylic acid

- 271 046336

Using the general procedure for amine substitution and hydrolysis starting from synthesis product 14 and morpholine as a suitable amine, the desired product was obtained.

MSHR-ERI (m/z): [M+H]+, calculated for C ₄₂ H ₅₂ N ₉ O ₄ S: 778.3863; found: 778.3858.

Example 271: 6-[[6-(1,3-Benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]methylamino]-3-[1-[[3-[2-(2carboxyethylamino)ethoxy]- 5,7-dimethyl-1-adamantyl]methyl]-5-methylpyrazol-4-yl]pyridine-2carboxylic acid

Using the general procedure for carrying out the amine substitution reaction and hydrolysis, starting from the synthesis product 14, 3-aminopropanoic acid as a suitable amine and 10 eq. K ₂ CO ₃ as a base gave the target product.

MSVR-ERI (m/z): [M+H] ⁺ , calculated for C41H50N9O5S: 780.3656; found: 780.3658.

Example 272: 6-[[6-(1,3-Benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]methylamino]-3-[1-[[3-[2-[(3hydroxyphenyl)methylamino] ethoxy]-5,7-dimethyl-1-adamantyl]methyl]-5-methylpyrazol-4-yl]pyridine-2carboxylic acid

Using the general procedure for amine substitution reaction and hydrolysis, starting from the synthesis product 14 and 3-(aminomethyl)phenol as a suitable amine, the desired product was obtained.

MSHR-ERI (m/z): [M+H] ⁺ , calculated for C ₄₅ H ₅₂ N ₉ O ₄ S: 814.3863; found: 814.3861.

The following Examples 273-280 were synthesized using a general amine displacement and hydrolysis procedure starting from synthesis 14 and a suitable amine.

Example 273: 6-[[6-(1,3-Benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]methylamino]-3-[1-[[3,5dimethyl-7-(2-pyrrolidin- 1-ylethoxy)-1-adamantyl]methyl]-5-methylpyrazol-4-yl]pyridine-2-carboxylic acid

Example 274: 6-[[6-(1,3-Benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]methylamino]-3-[1-[[3,5dimethyl-7-[2-(4 -methylpiperazin-1-yl)ethoxy]-1-adamantyl]methyl]-5-methylpyrazol-4-yl]pyridin-2carboxylic acid

- 272 046336

Example 275: 6-[[6-(1,3-Benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]methylamino]-3-[1-[[3-[2-(4hydroxybutylamino)ethoxy]- 5,7-dimethyl-1-adamantyl]methyl]-5-methylpyrazol-4-yl]pyridine-2carboxylic acid

Example 276: 6-[[6-(1,3-Benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]methylamino]-3-[1-[[3,5dimethyl-7-(2-piperazin- 1 -ylethoxy)-1 -adamantyl]methyl]-5-methylpyrazol-4-yl]pyridine-2-carboxylic acid

Example 277: 6-[[6-(1,3-Benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]methylamino]-3-[1-[[3-[2-[3hydroxypropyl(methyl)amino ]ethoxy]-5,7-dimethyl-1-adamantyl]methyl]-5-methylpyrazol-4-yl]pyridine-2carboxylic acid

Example 278: 6-[[6-(1,3-benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]methylamino]-3-[1-[[3-[2-[4hydroxybutyl(methyl)amino ]ethoxy]-5,7-dimethyl-1-adamantyl]methyl]-5-methylpyrazol-4-yl]pyridine-2carboxylic acid

- 273 046336

Example 279: 6-[[6-(1,3-Benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]methylamino]-3-[1-[[3-[2-[[3hydroxy-2- (hydroxymethyl)propyl]amino]ethoxy]-5,7-dimethyl-1-adamantyl]methyl]-5-methylpyrazol-4yl]pyridine-2-carboxylic acid

Example 280: 6-[[6-(1,3-benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]methylamino]-3-[1-[[3-[2[bis(3-hydroxypropyl) amino]ethoxy]-5,7-dimethyl-1-adamantyl]methyl]-5-methylpyrazol-4-yl]pyridine2-carboxylic acid

Example 281: 2-[[6-(1,3-Benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]methylamino]-5-[3-[2-fluoro4-(3-piperazin-1-ylprop -1-ynyl)phenoxy]propyl]thiazole-4-carboxylic acid

ABOUT

Example 282: 2-[[6-(1,3-Benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]methylamino]-5-[3-[2-fluoro4-[3-(4-methylpiperazin- 1 -yl)prop-1-ynyl]phenoxy]propyl]thiazole-4-carboxylic acid

- 274 046336

Example 283: 2-[[6-(1,3-Benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]methylamino]-5-[3-[2-fluoro4-[3-[3-hydroxypropyl( methyl)amino]prop-1-ynyl]phenoxy]propyl]thiazole-4-carboxylic acid

Example 284: 2-[[6-(1,3-benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]methylamino]-5-[3-[4-[3[[(3S)-3, 4-dihydroxybutyl]methylamino]prop-1-ynyl]-2-fluorophenoxy]propyl]thiazole-4-carboxylic acid

Example 285: 2-[[6-(1,3-benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]methylamino]-5-[3-[2-fluoro4-[3-(3-hydroxypropylamino) prop-1-ynyl]phenoxy]propyl]thiazole-4-carboxylic acid

Example 286: 2-[[6-(1,3-benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]methylamino]-5-[3-[4-[3[[(3S)-3, 4-dihydroxybutyl]amino]prop-1-ynyl]-2-fluorophenoxy]propyl]thiazole-4-carboxylic acid

Example 287: 2-[[6-(1,3-Benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]methylamino]-5-[3-[2-fluoro4-[3-[4-hydroxybutyl( methyl)amino]prop-1-ynyl]phenoxy]propyl]thiazole-4-carboxylic acid

- 275 046336

Example 288: 2-[[6-(1,3-benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]methylamino]-5-[3-[4-[3[[(3R)-3, 4-dihydroxybutyl]methylamino]prop-1-ynyl]-2-fluorophenoxy]propyl]thiazole-4-carboxylic acid

Example 289: 2-[[6-(1,3-benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]methylamino]-5-[3-[2-fluoro4-[3-(4-hydroxybutylamino) prop-1-ynyl]phenoxy]propyl]thiazole-4-carboxylic acid O

Example 290: 2-[[6-(1,3-benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]methylamino]-5-[3-[4-[3[[(3R)-3, 4-dihydroxybutyl]amino]prop-1-ynyl]-2-fluorophenoxy]propyl]thiazole-4-carboxylic acid

Pharmacological study

Example A: Fluorescence polarization analysis data.

Fluorescence polarization measures the rotation of fluorescent molecules in solution, with the larger the molecule, the more polarized the fluorescence emission.

The fluorescent PUMA probe (UniProtKB®, main accession number Q9BXH1 - SEQ ID: 01) fluorescein-beta-Ala-Ahx-AREIGAQLRRMADDLNAQY-OH from Biopeptides binds to GST(1-218)-(FACTQR_XA)-hsBCLXL(2- 209) having the amino acid sequence (SEQ ID: 02):

- 276 046336

[MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKKFELG LEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYD

ALDVVLYMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATF GGGDHPPK SDLIEGRGIPEFEF S Q SNREL V VDFL S YKL S QKGY S W S QF SD VEENR TEAPEGTESEMETPSAINGNPSWHLADSPAVNGATGHSSSLDAREVIPMAAVKQ ALREAGDEFELRYRRAFSDLTSQLHITPGTAYQSFEQ VVNELFRDGVNWGRIVA FFSFGGALCVESVDKEMQVLVSRIAAWMATYLNDHLEPWIQENGGWDTFVEL YGNNAAAESRKGQER] (GST UniProtKB®, main accession number P08515, and BCLXL UniProtKB®, main accession number Q07817-1), which leads to an increase in anisotropy. If a compound is added that competitively binds to the same site as the probe, thereby releasing it, the anisotropy is reduced due to an increase in the amount of free probe.

11-point serial dilutions of each compound were prepared in DMSO, the final buffer was as follows: 10 mM 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid [HEPES], 150 mM NaCl, 0.05% Tween 20, pH 7.4, and 5% DMSO . The final protein concentration for the assay was 20 nM with the fluorescent probe present at a concentration of 10 nM. Samples were incubated for 2 h at 23°C, after which fluorescence polarization was measured on a Biotek SynergyNeo plate reader (excitation 485 nm, emission 525 nm, parallel and perpendicular readings). Dose-response curves were then generated using XLFit software using a 4-parameter logistic model (sigmoidal dose-response model) and the inhibitory concentrations that resulted in a 50% increase in fluorescence intensity (IC ₅₀ ) were determined. KI values were determined from IC ₅₀ values according to Cer et al., Nucleic Acids Res, 2009, Jul 1;37(WebServer issue): W441-W445.

The results are summarized in table. 1. They show that the compounds of the present invention inhibit the interaction between the Bcl-xL protein and the fluorescent peptide described above.

- 277 046336

45 1.4E-08 46 1.9E-07 47 2.ZE-08 48 1.9E-06 49 8.1E-09 50 1.2E-09 51 6.4E-08 52 9.0E-08 53 1.5E-08 54 1.4E-09 55 8.2E-08 56 > 1E-05 57 3.5E-07 58 4.9E-08 59 < 1E-09 60 1.5E-09 61 < 1E-09 62 < 1E-09 63 < 1E-09 64 3.1E-08 65 < 1E-09 66 1.9E-09 67 < 1E-09 68 5.7E-09 69 4.2E-09 70 3.0E-08 71 > 1E-05 72 1.6E-07 73 < 1E-09 74 1.1E-09 75 < 1E-09 76 < 1E-09 170 < 1E-09 171 < 1E-09

77 1.1E-07 78 1.1E-07 79 9.2E-08 80 8.1E-08 81 1.6E-08 82 < 1E-09 83 3.2E-07 84 4.9E-08 85 < 1E-09 86 1.6E-07 87 5.ZE-07 88 2.8E-09 89 1.6E-07 90 < 1E-09 91 6.4E-09 92 6.8E-08 93 6.8E-09 94 6.7E-08 95 2.4E-08 96 2.2E-07 97 1.ZE-07 98 2.ZE-09 99 2.4E-06 100 1.6E-09 101 < 1E-09 102 2.2E-07 103 1.4E-08 104 1.6E-08 105 3.8E-07 106 2.7E-08 107 1.ZE-07 108 < 1E-09 172 < 1E-09

109 < 1E-09 119 < 1E-09 123 < 1E-09 124 < 1E-09 127 < 1E-09 130 < 1E-09 134 < 1E-09 138 2.9E-08 139 6.4E-07 140 < 1E-09 141 < 1E-09 142 5.0E-08 143 6.2E-08 144 < 1E-09 145 E8E-07 146 < 1E-09 149 EZE-07 150 < 1E-09 151 < 1E-09 156 8.9E-07 157 E2E-06 158 4.7E-08 159 E8E-07 160 2.2E-07 161 < 1E-09 163 < 1E-09 164 < 1E-09 165 < 1E-09 166 < 1E-09 167 < 1E-09 168 < 1E-09 169 < 1E-09

Example B: AlphaLISA assay data.

The AlphaLISA assay provides a measurement of the increase in alpha signal upon binding of biotinylated GST(l-218)-(FACTOR_XA)-hsBCLXL(2-2Q9) (seq id. 02 - _G s _T UniProtKB®, main accession number P08515, and BCLXL UniProtKB®, main accession number Q07817-1) attached to a streptavidin-labeled alpha donor bead, with an N-terminally labeled BIM-derived peptide (fluorescein-beta-Ala-Ahx-SEQ ID: 05, where SEQ ID: 05 is DMRPEIWIAQELRRIGDEANAYYARR) attached to an anti-FTIC AlphaLISA acceptor bead.

GST(l-218)-(FACTOR_XA)-hsBCLXL(2-2Q9) was biotinylated in 50 mM phosphate buffer using 20-fold excess EZ Link NHS-LC-biotin (thermoFisher, cat. no. 21336) overnight at 4°C .

The addition of a compound that competitively binds to the same site as the peptide will result in a decrease in the alpha signal of the accessible donor bead-protein-peptide-acceptor bead complex due to peptide displacement.

11-point serial dilutions of each compound were prepared in DMSO, final buffer

- 278 046336 The medium was as follows: 10 mM 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid [HEPES], 150 mM NaCl, 0.05% Tween 20, pH 7.4, and 4% DMSO. The final protein concentration in the assay was 20 pM with the peptide present at 1 nM. Alpha beads were used at a final concentration of 5 μg/ml. Samples were incubated overnight at 23°C after addition of acceptor beads. Finally, donor beads were added and incubated for 2 h at 23°C, after which the alpha signal was measured on a Biotek Synergy2 plate reader (excitation 680 nm, emission 615 nm). Dose-response curves were then generated using XLFit software using a 4-parameter logistic model (sigmoidal dose-response model) and the inhibitory concentrations that resulted in a 50% increase in fluorescence intensity (IC ₅₀ ) were determined.

The results are summarized in table. 2.

Example C: Attenuation analysis data.

The fluorescence quenching assay measures changes in the fluorescence intensity of the Cy5-labeled BCL-xL protein His-His-(EK)-hsBCLXL(2-197)[N197C] (UniProtKB®, main accession number Q07817-1), which has the amino acid sequence (SEQ ID: 03):

[MHHHHHHHHGATGSTAGSGTAGSTGASGASTGGTGATHHHHHHHDD DDKSPMGSQSNRELVVDFLSYKLSQKGYSWSQFSDVEENRTEAPEGTESEMETP SAINGNPSWHLADSPAVNGATGHSSSLDAREVIPMAAVKQALREAGDEFELRY RRAFSDLTSQLHITPGTAYQSFEQVVNELFRDGVNWGRIVAFFSFGGA LCVESV DKEMQVLVSRIAAWMATYLNDHLEPWIQENGGWDTFVELYG],

- 279 046336 which is linked in the C-terminal region to amino acid X, which corresponds to a cysteine labeled on sulfur with Sulfo-Cyanine5 from Lumiprobe GmbH, catalog number 13380, when binding a C-terminally labeled peptide derived from PUMA (UniProtKB®, main accession number Q9BXH1) having the amino acid sequence (SEQ ID: 04): [QWAREIGAQLRRMADDLNAQY], which is linked in the C-terminal region to the amino acid X', where X' is a cysteine labeled on sulfur with TQ5WS from AAT Bioquest, catalog number 2079.

The addition of a compound that competitively binds to the same site as the peptide will result in an increase in the fluorescence intensity of the protein due to the replacement of the fluorescence quencher.

11-point serial dilutions of each compound were prepared in DMSO, the final buffer was as follows: 10 mM 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid [HEPES], 150 mM NaCl, 0.05% Tween 20, pH 7.4, and 5% DMSO . The final protein concentration when performing the assay was 1 nM with the peptide present at a concentration of 400 nM. The samples were incubated for 2 h at 23°C, after which the fluorescence intensity was measured on a Biotek SynergyNeo plate reader (excitation 620 nm, emission 680 nm). Dose-response curves were then generated using XL-Fit software using a 4-parameter logistic model (sigmoidal dose-response model) and the inhibitory concentrations that resulted in a 50% increase in fluorescence intensity (IC ₅₀ ) were determined. _KI values were determined from IC ₅₀ values according to Cer et al., Nucleic Acids Res, 2009, Jul 1;37 (WebServer issue): W441-W445.

The results are summarized in table. 3.

Table 3

Example Ki(M) 165 5.9E-12 194 3.2E-12 50 4.4E-12 166 2.6E-12 195 1.8E-12 62 6.0E-12 167 2.9E-12 196 9.0E-13 73 2.8E-12 168 2.6E-12 197 1.ZE-12 74 6.1E-12 169 4.4E-12 198 7.3E-12 101 5.2E-13 170 5.0E-12 199 1.0E-12 108 5.9E-13 171 3.5E-12 200 7.7E-12 109 1.1E-12 172 2.6E-12 201 1.9E-12 114 1.1E-11 173 3.ZE-12 202 8.9E-13 116 8.6E-12 174 4.ZE-11 203 2.2E-12 117 6.6E-12 175 1.1E-12 204 6.0E-12 118 3.ZE-11 176 1.1E-12 205 2.ZE-12 119 5.5E-13 177 Z.OE-12 206 9.0E-12 122 Z.OE-12 178 7.4E-12 207 3.7E-12 123 8.4E-13 179 6.8E-12 208 8.8E-12 124 1.7E-12 180 5.2E-12 209 2.ZE-11 127 6.8E-13 181 4.ZE-12 210 8.5E-13 128 2.8E-11 182 7.4E-12 211 6.5E-12 130 3.7E-13 183 7.8E-12 212 1.0E-12 134 1.0E-12 184 3.6E-12 213 1.5E-12 141 5.5E-12 185 1.2E-12 214 4.0E-12 144 3.9E-12 186 1.0E-12 215 3.9E-12 146 1.2E-12 187 2.5E-12 216 4.0E-12 147 9.7E-12 188 1.8E-12 217 4.1E-12 148 1.2E-12 189 2.4E-12 218 1.2E-12 150 1.7E-12 190 3.2E-12 219 8.2E-13 151 8.5E-13 191 2.8E-12 220 3.5E-12 163 2.0E-11 192 2.9E-12 221 1.7E-12 164 1.9E-12 193 1.5E-12 222 1.4E-12

- 280 046336

223 1.8E-12 224 2.0E-12 225 3.5E-12 226 5.ZE-12 227 3.5E-12 228 2.2E-10 229 1.4E-12 230 1.4E-12 231 1.ZE-12 232 1.7E-12

233 4.ZE-12 234 1.6E-12 235 1.1E-12 237 1.8E-12 238 4.5E-12 239 2.7E-12 240 7.3E-12 241 9.4E-12 242 7.8E-12 243 5.0E-12

247 1.2E-12 250 2.2E-12 253 7.0E-11 267 1.6E-11 268 2.5E-12 269 2.7E-12 270 Z.OE-12 271 3.ZE-12 272 2.ZE-12

Table results 1-3 show that the compounds of the present invention are potent inhibitors of the Bcl-xL protein.

Example D: Study of the effect of Bcl-xL inhibitors on the survival of MOLT-4 or H146 cells using the MTT assay.

The MTT colorimetric assay is based on the mitochondrial reduction of tetrazolium salt in living cells. The number of viable cells is proportional to the production of formazan salts, which can be read spectrophotometrically at a wavelength of 540 nm.

MOLT-4 and H146 cells were purchased from ATCC and cultured in RPMI 1640 supplemented with 10% heat-inactivated fetal bovine serum, penicillin (100 IU/ml), streptomycin (100 mg/ml), and L-glutamine (2 mM). Cells were cultured at 37°C in a humidified atmosphere containing 5% CO ₂ . Cells were seeded in 96-microwell plates (150 μl per well) and exposed to compounds for 48 h (3.16-fold serially diluted; 9 concentrations of each, in triplicate). After the incubation time, 15 μl of MTT solution (5 mg/ml) per well was added and the cells were incubated for another 4 hours. Then 100 μl per well of 10% sodium dodecyl sulfate (SDS)/HCl 10 mM solution was added and the plate was incubated in overnight, after which the optical density was measured at a wavelength of 540 nm. IC ₅₀ values were calculated using standard four-parameter curve fitting. The IC ₅₀ value is defined as the concentration of the compound at which the MTT signal is reduced to 50% relative to the signal measured for the control. The results are the average values of at least 2 independent experiments and are presented in table. 4 below.

- 281 046336

Example N146 1C ₅₀ (M) MOLT-4 1С ₅₀ (M) 1 >1.5E-05 #nd 2 1.25E-05 #nd 3 4.2E-08 9E-08 5 8.21E-06 #ND 6 >=1.49E-05 #ND 7 >1.5E-05 #ND 8 1.05E-05 #ND 9 2.07E-07 2.32E-06 10 8.31E-06 #nd 12 9.13E-07 1.62E-06 13 2.92E-07 2.23E-06 14 2.11E-06 #nd 15 >1.5E-05 #nd 16 8.04E-06 #nd 17 1.34E-05 7.88E-06 18 >1.5E-05 #ND 19 >=1.28E-05 #ND 20 3.35E-06 7.21E-06 21 9.58E-06 #nd 22 2.13E-06 #nd 23 >=1.27E-05 1.ZE-05 24 6.93E-06 3.31E-06 25 3.18E-06 7.13E-06 26 >1.5E-05 >1.5E-05 27 >1.5E-05 >1.5E-05

Table 4

28 >1.5E-05 >1.5E-05 29 >1.5E-05 >1.5E-05 thirty >1.5E-05 8.67E-06 31 4.19E-06 2.34E-06 32 >=7.42E-06 1.33E-05 33 4.37E-06 6.25E-06 34 5.32E-06 3.89E-06 35 2.33E-06 3.7E-06 36 1.99E-06 3.47E-06 37 1.1E-05 7.39E-06 38 >1.5E-05 >1.5E-05 39 >1.5E-05 >1.5E-05 40 3.8E-06 1.49E-06 41 >=9.76E-06 >1.5E-05 42 2.04E-06 1.02E-06 44 >1.5E-05 >1.5E-05 45 >=1.18E-05 1.17E-05 46 >1.5E-05 >1.5E-05 47 >=9.62E-06 >1.5E-05 49 >1.5E-05 >1.5E-05 50 2.84E-09 4.62E-10 51 >1.5E-05 >1.5E-05 52 1.ZE-05 8.36E-06 53 1.65E-06 7.01E-07 54 3.52E-08 5.07E-09 56 >1.5E-05 >1.5E-05 57 1.34E-05 >1.5E-05 58 >1.5E-05 >1.5E-05

- 282 046336

59 2.71E-07 7.61E-08 60 5.8E-06 2.74E-06 61 5.93E-07 l,92E-07 62 4.66E-09 4.44E-10 63 2D5E-07 8.53E-08 64 2.24E-08 3.59E-06 65 8.65E-07 2.36E-07 66 1.16E-06 5.27E-07 67 7.1E-07 7.54E-07 68 2.6E-06 2.79E-07 69 6.86E-07 4.46E-07 70 >1.5E-05 >=1.16E-05 71 >1.5E-05 >1.5E-05 73 4.8E-09 1.51E-09 74 6.78E-09 6.72E-09 75 >1.5E-05 2.92E-06 76 1.13E-08 2.37E-09 77 >=1.49E-05 6.59E-06 78 >1.5E-05 >1.5E-05 82 9.88E-08 6.83E-08 83 >1.5E-05 >1.5E-05 84 >1.5E-05 7.57E-06 85 >1.5E-05 >1.5E-05 86 >=1.24E-05 1.46E-05 87 >1.5E-05 >1.5E-05 88 1.1E-05 1.61E-06 89 >1.5E-05 1.31E-05 90 2.01E-07 2.33E-08 91 5.39E-06 1.13E-06 92 >1.5E-05 >1.5E-05 93 >1.5E-05 9.4E-06 94 4.07E-06 >=1E-O5

95 9.27E-06 5.01E-06 98 >1.5E-05 >1.5E-05 100 3.86E-08 1.26E-08 101 1.19E-06 4.83E-07 102 >1.5E-05 >1.5E-05 105 >1.5E-05 >1.5E-05 106 >1.5E-05 >1.5E-05 108 2.23E-08 4.67E-08 109 6.38E-09 1.98E-08 112 3.09E-08 7.75E-08 113 1.3E-08 2.57E-08 114 1.58E-09 3.81E-09 115 >1.5E-07 >1.5E-07 116 1.91E-08 3.06E-09 117 4.96E-08 4.86E-07 118 3.26E-08 0.000000096 119 2.14E-08 9.74E-08 120 2.8E-07 5.57E-07 122 5.14E-08 0.00000028 123 6.25E-09 1.65E-08 124 5.06E-08 1.92E-08 127 9.04E-09 1DZE-08 128 2.08E-08 l,81E-08 130 2.01E-07 l,08E-07 134 1.91E-09 7.83E-10 135 5.29E-06 3.24E-06 137 3.2E-07 6.14E-08 140 1.42E-07 2.34E-08 144 1.45E-07 4.3E-07 146 9.71E-07 6.68E-07 150 1.68E-08 1.36E-07 151 #ND 1.94E-07

- 283 046336

161 3.02E-08 3.23E-09 162 6.59E-06 3.17E-06 163 1.04E-06 9.62E-08 164 3.46E-08 1.29E-08 165 2.11E-07 4.ZE-07 166 1.42E-07 1.72E-07 167 4.32E-07 4E-07 168 #nd 1.78E-07 169 7.91E-08 1.06E-07 170 1.56E-07 1.54E-07 171 1.1E-07 4.11E-07 172 4.02E-08 1.6E-07 173 2.08E-07 1.27E-07 174 1.26E-05 1.66E-06 175 7.5E-08 2.04E-07 176 1.24E-07 7.62E-08 177 3.63E-08 0.000000225 182 1.99E-07 1.29E-07 183 1.87E-07 4.65E-08 184 1.07E-07 1.84E-07 185 7E-09 1.68E-08 186 1.69E-08 8.53E-08 187 2.34E-07 1.03E-07 188 5.44E-09 3.41E-08 189 9.48E-08 1.71E-07 190 7.83E-08 5.49E-08 191 8.38E-08 2.2E-06 192 1.19E-08 9.64E-08 193 6.5E-08 9.25E-08 194 2.7E-08 1.29E-07 195 5.25E-07 5.29E-07 196 L62E-06 8.79E-07

197 4.39E-08 #nd 198 1.42E-08 1.59E-09 199 6.41E-08 2.59E-08 200 1.15E-07 6.95E-07 201 7.53E-08 4.17E-07 202 7.48E-08 6.11E-07 203 3.9E-09 7.53E-10 204 7.ZE-09 7.03E-10 205 <=5.93E-09 3.74E-10 206 1.5E-08 1.48E-09 207 1.08E-06 3.28E-06 208 7.33E-09 7.53E-10 210 2.14E-08 2.03E-08 211 6.68E-08 4.84E-07 212 3.24E-06 2.88E-06 213 7.28E-08 9.36E-08 214 2.67E-08 2.25E-08 215 1.74E-07 4.06E-07 216 2.22E-07 1.46E-07 217 8.62E-08 1.8E-07 218 8.91E-07 2.7E-07 219 1.29E-07 1.27E-06 220 2.21E-07 9.84E-07 221 9.44E-08 2.95E-08 222 3.24E-08 6.1E-08 223 7.46E-09 3.13E-09 224 1.16E-06 2.18E-06 225 3.51E-07 1.08E-06 226 #nd 7.52E-07 227 #nd 8.02E-07 228 >ZE-05 #nd 229 4.37E-07 1.64E-06

- 284 046336

230 1.38E-06 2.18E-06 231 1.61E-06 1.19E-06 233 2.55E-09 8.79E-10 234 1.5E-08 3.8E-09 235 4.13E-07 4.13E-07 237 #nd 7.5E-07 238 #nd 1.49E-07 239 #nd 3.14E-07 240 #nd 1.32E-06 241 #nd 1.19E-06 242 #nd 5.25E-07

243 #ND 1.12E-07 247 #nd 2.81E-09 250 #nd 2.46E-10 253 #nd 3.17E-08 267 #nd 4.38E-08 268 #nd 2.55E-10 269 #nd 1.82E-10 270 #nd 1.99E-10 271 #nd 3.08E-08 272 #nd 2.84E-10

These data indicate that most of these compounds are active in cells and can induce a dose-dependent reduction in the survival of H146 and Molt-4 cell lines.

Example E: pharmacodynamics and tumor regression study.

The in vivo therapeutic and pharmacodynamic effects of small molecules targeting Bcl-xL were determined in the MOLT-4 T-cell acute lymphoblastic leukemia (T-ALL) model when administered intravenously (IV). Test compounds were formulated with 20% hydroxypropyl cyclodextrin/25 mM hydrogen chloride (HPBCD/HCl).

Materials and methods.

MOLT-4 cells (ATCC no. CRL-1582) were cultured in RPMI supplemented with 10% FBS. Cells were resuspended in 50% Matrigel (BD Biosciences), and 0.1 ml of a suspension containing 5 × 10 ⁶ cells was inoculated subcutaneously into the right flank of female NOD SCID mice (Charles River). For efficacy studies, once tumors had reached an appropriate volume, mice were randomized (8 animals per group) using Easy stat software. Vehicle control (HPBCD/HCl), Example 73 (3.75 or 7.5 mg/kg) or Example 100 (3.75 mg/kg) was administered IV (twice weekly for 3 weeks - Q3D6). The body weight of the mice was monitored three times a week and tumor size was measured using an electronic caliper. Tumor volume was assessed by measuring the minimum and maximum tumor diameters and calculating using the formula: (minimum diameter) (maximum diameter)/2. At the end of the treatment cycle (day 17), tumor growth inhibition was calculated using the formula:

/ Median value (TV per Dx in the treatment group) \ \ Median value (TV per Dx in the control group)/

Response was assessed as follows: CR (complete response) if the tumor size was <25 mm ³ for at least three consecutive measurements, PR (partial response) if the tumor size was between 25 mm ³ and half the initial size for at least three sequential measurements. Mice were sacrificed at the first measurement in which the tumor volume exceeded 2000 mm ³ or at the first sign of deterioration in animal health.

For pharmacodynamic studies, after tumors had reached an appropriate volume, mice were randomized (3 animals per group) using Easy stat software. Vehicle control (HPBCD/HCl) or small molecules targeting Bcl-xL were administered IV (once daily QD) at a dose of 7.5 mg/kg. Tumor samples were collected 6 hours after dosing and lysed (10 mM HEPES pH 7.4, 142.5 mM KCl, 5 mM MgCl ₂ , 1 mM EDTA, 1% NP40, protease and phosphatase inhibitor cocktails - Calbiochem). Cleared lysates were prepared for immunodetection of cleaved PARP and cleaved caspase 3 using the MSD apoptosis panel whole cell lysate kit (MSD), in 96-well plates according to the manufacturer's instructions, and analyzed on a QuickPlex SQ 120 instrument. Whole blood samples were analyzed on the instrument Hematology Analyzer Coulter Ac-T diff (Beckman Coulter).

All experiments were carried out in accordance with current French regulations after approval by the ethics committee of the Servier Research Institute (IdRS). NOD SCID mice were housed according to established protocols.

Results.

The effectiveness of the compound of example 73 and the compound of example 100 on MOLT-4 xenografts is illustrated in table. 5. Treatment was started 12 days after inoculation with tumor cells (average size: 214 mm ³ ). Vehicle (HPBCD/HCl), Example 73 (3.75 and 7.5 mg/kg) or Example 100 (3.75 mg/kg) were dosed IV every 3 days for a total of 6 administrations.

On day 17 after initiation of treatment, tumor growth inhibition (%TGI) for compound Example 73

- 285 046336 was 56.9% at a dosage of 3.75 mg/kg and 83.3% at a dosage of 7.5 mg/kg (p < 0.05), as shown in table. 5. At the same time point, the %TGI induced by Example 100 at 3.75 mg/kg was 64.6% (p<0.05).

No clinically significant weight loss was observed as a result of treatment.

Table 5

Inhibition of MOLT-4 Tumor Growth by Treatment with Example 73 (3.75 and 7.5 mg/kg IV, Q3D6) or Example 100 (3.75 mg/kg IV, Q3D6)

Connection under test Dose (mg/kg) %TGI (day 17) Example 73 3.75 56.9* Example 73 7.5 83.3* Example 100 3.75 64.6*

*p value <0.05 compared with the control group.

The effect of small molecules targeting Bcl-xL on the induction of apoptosis in MOLT-4 tumor cells and the number of circulating platelets is illustrated in Table 1. 6. Treatment was started 18 days after tumor cell inoculation (mean size: 349 mm ³ ). Vehicle (HPBCD/HCl), compounds of Example 74, Example 76 and Example 73 (7.5 mg/kg) were dosed once IV, and samples were collected after 6 hours. All compounds showed the induction of markers of apoptosis, namely cleaved PARP (at 14.226 .6-fold interval compared to control), and cleavage by cleavage caspase 3 (in 4.7-5.1-fold interval compared to control). In addition, given the well-described role of Bcl-xL in regulating platelet lifespan, all tested compounds caused a significant reduction in platelet counts (up to 2-4% of control values).

Table 6

Cleaved PARP and cleaved caspase 3 in tumor cells and platelet loss in female NOD SCID mice transplanted with MOLT-4 6 hours after treatment with 7.5 mg/kg of these compounds administered ^i.v.

Connection under test Dose (mg/kg) and route Point in time (h) Cleaved PARP (fold increase compared to control) Cleavage caspase 3 (fold increase compared to control) Platelet count (x10 ³ / μl) Remaining platelets (compared to vehicle), % Example 74 7.5, i/v 6 17.1 5.1 18 2 Example 76 7.5, i/v 6 14.2 4.9 40 3 Example 73 7.5, i/v 6 21.4 4.7 51 4

In conclusion, this study shows that the small molecules targeting Bcl-xL, which are described in Examples 73, 74, 76 and 100, are active in vivo after intravenous administration. Tumor regression, induction of apoptosis in tumor cells and a strong reduction in the number of circulating platelets were observed, which is consistent with the previously described role of Bcl-xL in the control of apoptosis and regulation of platelet lifespan (Youle and Strasser, Nat. Rev. Mol. Cell Biol. 2008 Jan; 9(1):47-59; Zhang et al., Cell Death Differ. 2007 May; 14(5):943-51; Mason et al., Cell 2007 Mar 23; 128(6):1173-86). In addition, no clinically significant weight loss was observed with treatment at effective doses, and platelet loss was reversed after cessation of treatment (data not shown). Overall, these data indicate that there is an acceptable therapeutic window for the use of these small molecules targeting Bcl-xL in the treatment of cancer.

Example F: in vivo pharmacodynamic profile of the compound of formula (I).

The pharmacokinetic profile of compounds of formula (I) was assessed in rodents (mice, rats) after administration by p/o and/or i.v. route. The composition was selected based on the physicochemical properties of the drug being tested, as well as the route of administration. A single dose of the drug (<5 mg/kg), prepared in an adapted formulation, was administered to animals via the IV (bolus or 10-minute infusion) or p/o (gastric tube) route (3 animals/route). Blood samples from each animal (up to 6 samples/animal) were collected within 24 hours of dosing, and plasma test compound concentrations were determined by extraction followed by liquid chromatography coupled to tandem mass spectrometric detection (LC/MS-MS). .

In some cases, to determine the pharmacokinetic profile of compounds of this

- 286 046336 inventions in Wistar rats the following experimental protocol was used:

The drug was prepared as a formulation comprising a mixture of polyethylene glycol 300/anhydrous ethanol/NaCl 0.9% (40/10/50 v/v/v). The drug was administered intravenously to male Wistar rats (3 animals) at a dose of 0.75 mg/kg (10 min infusion, 5 ml/kg). Blood samples were collected from each animal at the following time points: completion of infusion (10 min), 0.5 h, 1 h, 3 h, 6 h, and 24 h postdose. Plasma test compound concentrations were determined by extraction followed by liquid chromatography coupled to tandem mass spectrometric detection (LC/MS-MS).

The lower limit of quantitation was 2.5 ng/ml.

The results allow the compounds of the present invention to be ranked based on their plasma concentration, elimination rate constant, clearance and volume of distribution in order to assess the therapeutic range of these compounds in an animal model.

Claims (42)

1. Compound of formula (I) where R1 and R2 independently of each other represent a group selected from hydrogen; linear or branched C1-C ₆ -alkyl, optionally substituted with a hydroxyl or C1-C ₆ -alkoxy group; C ₃ -C ₆ -cycloalkyl; trifluoromethyl; linear or branched C ₁ -C ₆ alkylene heterocycloalkyl, where the heterocycloalkyl group is optionally substituted with a linear or branched C 1 -C ₆ alkyl group; or R1 and R2 together with the carbon atoms they carry form a C ₃ -C ₆ -cycloalkylene group, R ₃ represents a group selected from hydrogen; C ₃ -C ₆ -cycloalkyl; linear or branched C _g Sb-alkyl; -X _r NRaRb; -X1-N ⁺ RaRbR _c ; -X1-O-Rc; -Xi-COORc; -X _r PO(OH)2; -X _r SO2(OH); -X1-N3 -Xi—=CH and , Ra and Rb independently of each other represent a group selected from hydrogen; heterocycloalkyl; linear or branched C1-C ₆ -alkyl, optionally substituted with one or two hydroxyl groups; C1-C ₆ -alkylene^O ₂ OH; C1-C ₆ -alkylene^O ₂ O ^- ; C1-C6-alkylene-COOH; C1-C6alkylene-PO(OH)2; C1-C6-alkylene-NRdRe; C1-C6-alkylene-N ⁺ RdR _e Rf; or Ra and R _b together with the nitrogen atom carrying them form the B1 ring; or Ra, Rb and Rc together with the nitrogen atom carrying them form a bridged C ₃ -C8 heterocycloalkyl, Rc, Rd, Re, Rf independently of each other represent hydrogen or a linear or branched C1-C ₆ alkyl group, or Rd and Re together with the nitrogen atom bearing them form the B ₂ ring, or Rd, Re and Rf together with the supporting nitrogen atom their nitrogen atom forms a bridged C ₃ -C8 heterocycloalkyl, Het1 represents the following group: - 287 046336 A1 means S, A2 means CH, G is selected from the group consisting of -C(O)ORG3, -C(O)NR _G1 R _G2 , -C(O)R _G2 , -NR _G1 C(O)R _G2 , RG1 and R _G2 in each case are each independently selected from the group consisting of hydrogen, C1-C6 alkyl, optionally substituted with 1-3 halogen atoms, ^-^-alkenyl, ^-^-alkynyl, C3-C6 cycloalkyl, phenyl and -(CH ₂ ) _1-4 -phenyl; RG3 is selected from the group consisting of ^-^-alkyl, optionally substituted with 1-3 halogen atoms, ^-^-alkenyl, ^-^-alkynyl, ^-^-cycloalkyl, phenyl and -(CH ₂ ) _1-4 - phenyl; or RG1 and RG2, together with the atom to which each is attached, combine to form ^-C^ heterocycloalkyl; or, alternatively, G is selected from the group consisting of: where RG4 is selected from ^-^-alkyl, optionally substituted with 1-3 halogen atoms, C2-C6 alkenyl, ^-^-alkynyl and ^-^-cycloalkyl, R ₄ represents hydrogen, a fluorine, chlorine or bromine atom, a methyl, hydroxyl or methoxy group, R5 represents a group selected from ^-^-alkyl, optionally substituted with 1-3 halogen atoms; ^-^-alkenyl; ^-^-alkynyl; halogen or -CN, R6 is a group selected from: hydrogen; - ^-^-alkenyl; - X2-O-R7; - X2-NSO2-R7; - C=C(R9)-Yi-O-R7; ^-^-cycloalkyl; C ₃ -C ₆ -heterocycloalkyl, optionally substituted with a hydroxyl group; ^-^-cycloalkylene^^; C ₃ -C ₆ -heterocycloalkylene-Y ₂ -R ₇ group, heteroarylene^ ₇ group, optionally substituted by a linear or branched ^-^-alkyl group, R ₇ represents a group selected from: a linear or branched ^-^-alkyl group; (C ₃ -C ₆ )cycloalkylene-R ₈ ; or: - 288 046336 where Su represents C ₃ -C8-cycloalkyl, R8 represents a group selected from hydrogen; linear or branched C1-C6 alkyl, -NR'aR'b;-NR'a-CO-OR'c;-NR'a-CO-R'c; -N ⁺ R'aR'bR'c;-O-R'c;-NH-X'2-N ⁺ R'aR'bR'c;-OX^-NR'aR'b,-nr;-x' ₉ —ch -X' ₂ -NR' _a R' _b , -NR' _c -X' ₂ -N ₃ and , R9 represents a group selected from linear or branched C1- _C6 alkyl, trifluoromethyl, hydroxyl, halogen, C1- _C6 alkoxy, R ₁₀ represents a group selected from hydrogen, fluorine, chlorine, bromine, -CF3 and methyl, R11 represents a group selected from hydrogen, C1- _C3 -alkylene^8, -O-C1-C3 _-alkylene - _K8 , -CO-NRhRi _and -C^CH-CrC^alkylene-L^, - CH=CH-CHO, C ₃ -C ₈ -cycloalkylene-CH ₂ -R ₈ , C3-C8heterocycloalkylene-CH ₂ -R ₈ , R12 and _R13 independently represent a hydrogen atom or a methyl group, _R14 and R15 independently represent hydrogen or a methyl group, or R14 and R15 together with the carbon atom they carry form cyclohexyl, R _h and R _i independently of each other represent hydrogen or a linear or branched C1-C ₆ alkyl group, X1 and X2 independently represent a linear or branched C1-C6 alkylene group, optionally substituted with one or two groups selected from trifluoromethyl, hydroxyl, halogen, C1- _C6 alkoxy, X' ₂ is a linear or branched C1-C ₆ -alkylene, R'a and R'b independently of each other represent a group selected from hydrogen; heterocycloalkyl; -SO ₂ -phenyl, where phenyl may be replaced by linear or branched C1-C ₆ -alkyl; linear or branched C1-C ₆ -alkyl, optionally substituted with one or two hydroxyl or C^-CD-alkoxy groups; C ₁ -C ₆ -alkylene-SO ₂ OH; C ₁ -C ₆ -alkylene-SO ₂ O ^- ; C1-C ₆ -alkylene-COOH; C1C ₆ -alkylene-PO(OH) ₂ ; C1-C ₆ -alkylene-Na^ _e ; CrC^alkylene^+Ka^^; C1-C6-alkylene-O-C 1 -C ₆ -alkyleneOH; C1-C ₆ -alkylenephenyl, where phenyl may be replaced by a hydroxyl or C1-C ₆ -alkoxy group; groups: or R' _a and R'b together with the nitrogen atom they carry form a B3 ring, or R' _a , R' _b and R' _c together with the nitrogen atom they carry form a bridged C ₃ -C ₈ -heterocycloalkyl, R' _c , R' _d , R' _e , R' _f independently of each other represent hydrogen or a linear or branched C1-C ₆ alkyl group, or R'd and R'e, together with the nitrogen atom carrying them, form a ring In ₄ , or R'd, R'e and R'f, together with the nitrogen atom bearing them, form a bridging C ₃ -C ₈ -heterocycloalkyl, Y1 is a linear or branched C1-C ₄ -alkylene, Y2 represents the bond, -O-, -O-CH2-, -O-CH)-, -O-SO2-, -CH2-, -CH2-O, -CH2-CO-, -CH ₂ -SO ₂ - , -C2H5-, -CO-, -TO-O-, -CO-CH2-, -OT^-CHg, -SO2-, -SO^Hr, -NH^O-, -NH-SO2-, m= 0, 1 or 2, - 289 046336 p=1, 2, 3 or 4, B1, _B2 , _B3 and _B4 independently represent a _C3 -C8 heterocycloalkyl group, which: (i) may be a mono- or bicyclic group, where the bicyclic group includes a fused, bridged or spiro ring system, ( ii) may contain, in addition to the nitrogen atom, one or two heteroatoms independently selected from oxygen, sulfur and nitrogen, (iii) may be substituted by one or two groups selected from: fluorine, bromine, chlorine, linear or branched ^- ^-alkyl, hydroxyl, -NH2, oxo or piperidinyl, its enantiomers and diastereoisomers, and their addition salts with a pharmaceutically acceptable acid or base, where heterocycloalkylene means divalent heterocycloalkyl, where heterocycloalkyl means a mono- or bicyclic non-aromatic carbocyclic group consisting of 3- 10 ring members, and containing from 1 to 3 heteroatoms selected from oxygen, sulfur, SO, SO ₂ and nitrogen, where the bicyclic group may be of fused or spiro ring type, heteroarylene means divalent heteroaryl, where heteroaryl means a mono- or bicyclic group consisting of 5-10 ring members, having at least one aromatic moiety and containing from 1 to 4 heteroatoms selected from oxygen, sulfur and nitrogen. 2. A compound according to claim 1, where Het1 is: 3. A compound according to claim 1 or 2, where R ₁ represents a methyl group and R 2 represents a hydrogen atom. 4. A compound according to claim 1 or 2, wherein each of R ₁ and R 2 represents a methyl group or a hydrogen atom. 5. A compound according to claim 1 or 2, wherein R1 or R2 is a linear or branched C1-C6 alkyl, optionally substituted with a hydroxyl or ^-^-alkoxy group, preferably a group selected from methyl, methoxymethyl, hydroxymethyl, ethyl and prop-2 -ila. 6. A compound according to claim 1 or 2, wherein R1 or R2 represents a C ₃ -C ₆ -cycloalkyl, preferably a cyclopropyl group. 7. A compound according to claim 1 or 2, where R1 represents a trifluoromethyl group. 8. A compound according to claim 1 or 2, wherein R2 represents a linear or branched C1-C6 alkylene heterocycloalkyl, preferably (4-methylpiperazin-1-yl)propyl group. 9. A compound according to claim 1 or 2, where R1 and R2 together with the carbon atoms they carry form a cyclohexene or cyclopentene group. 10. A compound according to any one of claims 1 to 9, where R ₃ represents a hydrogen atom or a methyl group. 11. A compound according to any one of claims 1 to 9, where R ₃ represents -X1-PO(OH) ₂ , -X1-SO ₂ (OH), -X1NRaRb; -X ₁ -N ⁺ RaRbRc, where Ra or Rb, or both of them, represent a group selected from C1-C6αalkylene-SO2OH, C1-C6-alkylene-SO2O ^- and C1-C6- _αalkylene -PO(OH) ₂ . 12. A compound according to any one of claims 1 to 9, where R8 represents -NR'aR'b; -N ⁺ R'aR'bR'c;-NH-X'2N ⁺ R'aR'bR'c, where R'a and R'b, or both of them, represent a group selected from ^-^-alkyleneSO2OH and C1-C6-alkylene-PO(OH )2. 13. A compound according to any one of claims 1 to 9, where R ₃ represents a group selected from ethyl, propyl; 2-methoxyethyl, 2-(morpholin-4-yl)ethyl, 3-(morpholin-4-yl)propyl, 3-diethylaminopropyl, 3-methoxypropyl, 3-hydroxypropyl, 3,4-dihydroxybutyl, 4-methoxy-3-hydroxybutyl, 4-hydroxy-3methoxybutyl, 2,3-dihydroxypropyl, 4,5-dihydroxypentyl, 4-hydroxybutyl, 3-hydroxy-2morpholinopropyl, 5-hydroxy-4-methoxypentyl, 5-morpholino-4-hydroxypentyl, 3-hydroxy-2methoxypropyl, 5-[2-(dimethylamino)ethylmethylamino]-4-hydroxypentyl, 5-hydroxypentyl, 5-methoxy-4-hydroxypentyl, 5-(dimethylamino)-4-hydroxypentyl, 4-hydroxy-5-(trimethylammonio)pentyl, 5-[3 -sulfonatepropyldimethylammonio]-4-hydroxypentyl, 4-hydroxy-5(methylamino)pentyl, 3-carboxypropyl, 5-[methyl(4-piperidyl)amino]pentyl, 5-(2morpholinoethylamino)pentyl, 5-[2-(4- methylpiperazin-1-yl)ethylamino]pentyl, 4-[2-(4methylpiperazin-1-yl)ethylamino]butyl, 4-hydroxy-5-[methyl-[2-(methylamino)ethyl]amino]pentyl, 5-[ 2(diethylamino)ethylamino]-4-hydroxypentyl, 5-(4-amino-1-piperidyl)-4-hydroxypentyl, 4-hydroxy5-piperazin-1-ylpentyl, 5-[2-(1-piperidyl)ethylamino]pentyl , 4-(4-amino-1-piperidyl)butyl, 4-[2(diethylamino)ethylamino]butyl, 5-(4-amino-1-piperidyl)pentyl, 4-[methyl-[2(methylamino)ethyl] amino]butyl, 4-(2-morpholinoethylamino)butyl, 4-[2-(4-methylpiperazin-1- 290 046336 yl)ethylamino]butyl, 4-[(1-methyl-4-piperidyl)amino]butyl, 5 -[2-(diethylamino)ethylamino]pentyl, 4-piperazin-1 -ylbutyl, 4-(methylamino)butyl, 5-piperazin-1 -ylpentyl, 5 - [methyl-2-(methylamino)]ethyl, 5-aminopentyl, 4 -aminobutyl, 5-(methylamino)pentyl, 5-[3-(4-methylpiperazin-1yl)propylamino]pentyl, 4-hydroxy-5-[methyl(2-phosphonoethyl)amino]pentyl, 6(dimethylamino)hexyl, 3 -(dimethylamino)propyl, 2-(trimethylammonio)ethyl, 3(trimethylammonio)propyl, 2-(dimethylamino)ethyl, 4-(dimethylamino)butyl, 5-[3sulfonatepropyldimethylammonio]pentyl, 4-(trimethylammonio)butyl, 4-hydroxy -5-quinuclidin-1-1-yl-pentyl, 4-hydroxy-5-(1-aza-4-azoniabicyclo[2.2.2]octan-4-yl)pentyl, 4-hydroxy-5-(4methylmorpholine-4- ium-4-yl)pentyl, 5-(trimethylammonio)pentyl, 4-hydroxy-5-(1-methylpiperidin-1 ium-1-yl)pentyl, 5-(1,4-dimethylpiperazin-1-ium-1 - yl)-4-hydroxypentyl, 6-(trimethylammonio)hexyl, 5-[3-hydroxypropyl(dimethyl)ammonio]pentyl, 5-[2-hydroxyethyl(dimethyl)ammonio]pentyl, 5[carboxymethyl(dimethyl)ammonio]pentyl, 5-[carboxymethyl(dimethyl)ammonio]pentyl, 3-phosphonopropyl, 3-[3-sulfonatepropyldimethylammonio]propyl, 3-sulfopropyl, 4-sulfobutyl, 4-phosphonobutyl, 4-carboxybutyl, 3-aminopropyl, 3-azidopropyl, pent-4-yne- 1-yl, 4-aminobutyl, 4azidobutyl, hex-5-yn-1-yl, 5-azidopentyl. 14. The compound according to any one of claims 1 to 13, where Het ₂ represents 15. A compound according to any one of claims 1 to 13, where Het ₂ represents coon 16. The compound according to claim 14 or 15, where R6 represents a hydrogen atom. 17. The compound according to claim 14, where R6 represents a group -X ₂ -OR ₇ , where X ₂ means a propylene group. 18. The compound according to claim 17, where R7 represents the following group: 19. The compound according to claim 17, where R7 represents the following group: 20. A compound according to claim 18 or 19, where R8 represents a group selected from dimethylamino, methylamino, methylethylamino, diethylamino, methyl[2-(methylamino)ethyl]amino, (2hydroxyethyl)(methyl)amino, 4-morpholinyl, pyrrolidine -1-yl, 1-piperidyl, [(tert-butoxy)carbonyl]-(methyl)amino, hydroxyl, bis(3-sulfopropyl)amino, 3-sulfopropylamino, methyl(3-sulfopropyl)amino, methyl(p-tolylsulfonyl)amino , (4-methoxyphenyl)methylmethylamino, 2(dimethylamino)ethylamino and 21. A compound according to claim 18 or 19, where R8 represents a group selected from: 3-piperazin-1yl, 4-methylpiperazin-1-yl, 3-hydroxypropyl(methyl)amino, [^)-3,4-dihydroxybutyl ]methylamino, 3-hydroxypropylamino, [(3S)-3,4-dihydroxybutyl]amino, 4-hydroxybutyl(methyl)amino, [(3R)-3,4dihydroxybutyl]methylamino, 4-hydroxybutylamino, [(3R)-3, 4-dihydroxybutyl]amino. 22. The connection according to claim 17, where R7 is: - 291 046336 where R11 is selected from dimethylcarbamoyl, 3-(dimethylamino)propyl, 3-(methylamino)propyl, 3(methylamino)propyl. 23. A compound according to claim 14, where R6 represents a C ₃ -C ₆ -heterocycloalkylene-Y ₂ -R ₇ group, where the heterocycloalkylene group is selected from: 24. A compound according to claim 23, wherein R7 is selected from methyl, methylethyl, tert-butyl, 2-methylpropyl and phenyl, preferably phenyl. 25. The compound according to claim 14 or 15, where R6 represents -C=C(R ₉ )-Y1-OR ₇ , where Y1 denotes a methylene group. 26. The connection according to claim 14 or 15, where R6 is: where R8 represents a group selected from hydrogen, 2-(methylamino)ethoxy and 28. The compound according to claim 26, where R7 represents a group selected from: where R ₈ represents a group selected from 2-(dimethylamino)ethoxy, 2-[(2sulfoethyl)amino]ethoxy, 2-[methyl(2-sulfoethyl)amino]ethoxy, 2-(3-hydroxypropylamino)ethoxy, 2( 3-methoxypropylamino)ethoxy, 2-morpholinoethoxy, 2-(2-carboxyethylamino)ethoxy, 2-[(3hydroxyphenyl)methylamino]ethoxy, 2-(methylamino)ethoxy, 2-pyrrolidin-1-ylethoxy, 2-(4methylpiperazine-1 -yl)ethoxy, 2-(4-hydroxybutylamino)ethoxy, 2-piperazin-1-ylethoxy, 2-[3hydroxypropyl(methyl)amino]ethoxy, 2-[4-hydroxybutyl(methyl)amino]ethoxy; 2-[[3-hydroxy-2(hydroxymethyl)propyl]amino]ethoxy; 2-[bis(3-hydroxypropyl)amino]ethoxy. 29. A compound according to claim 1, selected from the following group of compounds: 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(4-hydroxybutyl)amino)-5-(3-{2 - 292 046336 fluoro-4-[3 -(methylamino)prop-1 -in-1-yl]phenoxy}propyl)-1,3-thiazol-4-carboxylic acid, 2-[[6-( 1,3-benzothiazol-2-ylamino)-5 -methylpyridazin-3 -yl]methylamino] -5-[3-[4-[3(dimethylamino)prop-1 -inyl] -2 -fluorophenoxy]propyl]thiazole-4-carboxylic acid, 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)-5-(3-{2-fluoro-4-[3( methylamino)prop-1-yn-1-yl]phenoxy}propyl)-1,3-thiazol-4-carboxylic acid, 2-[[6-(1,3-benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]-(3,4-dihydroxybutyl)amino]-5-[3-[2fluoro-4-[3 - (methylamino)prop-1-ynyl]phenoxy]propyl]thiazole-4-carboxylic acid, 2-[[6-(1,3-benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]-(3-hydroxy-4-methoxybutyl)amino]-5 [3-[2-fluoro-4- [3-(methylamino)prop-1-ynyl]phenoxy]propyl]thiazole-4-carboxylic acid, 2-[[6-(1,3-benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]-(2,3-dihydroxypropyl)amino]-5-[3-[2fluoro-4-[3 - (methylamino)prop-1-ynyl]phenoxy]propyl]thiazole-4-carboxylic acid, 2-[[6-(1,3-benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]-(4,5-dihydroxypentyl)amino]-5-[3-[2fluoro-4-[3 - (methylamino)prop-1-ynyl]phenoxy]propyl]thiazole-4-carboxylic acid, 2-[[6-(1,3-benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]-(3,4-dihydroxybutyl)amino]-5-[3-[4-[3(dimethylamino) prop-1-ynyl]-2-fluorophenoxy]propyl]thiazole-4-carboxylic acid, 2-[[6-( 1,3-Benzothiazol-2-ylamino)-5 -methylpyridazin-3 -yl] -(3 -hydroxypropyl)amino] -5-[3-[2fluoro-4-[3 -(methylamino )prop-1-ynyl]phenoxy]propyl]thiazole-4-carboxylic acid, 2-[[6-(1,3-benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]-[4-hydroxy-5(trimethylammonio)pentyl]amino]-5-[3-[2-fluoro -4-[3-(methylamino)prop-1-ynyl]phenoxy]propyl]thiazole 4-carboxylate, 3-[[5-[[6-(1,3-benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]-[4-carboxy-5-[3-[2-fluoro-4-[3 (methylamino)prop-1-ynyl]phenoxy]propyl]thiazol-2-yl]amino]-2hydroxypentyl]dimethylammonio]propane-1-sulfonate, 2-[[6-(1,3-benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]-(4-hydroxybutyl)amino]-5-[3-[2-fluoro4-[3-[methyl (3-sulfopropyl)amino]prop-1-ynyl]phenoxy]propyl]thiazole-4-carboxylic acid, 5-{1-[(adamantan-1-yl)methyl]-5-methyl-1H-pyrazol-4-yl}-2-({6-[(1,3-benzothiazol-2-yl)amino]- 5methylpyridazin-3-yl}(methyl)amino)-1,3-thiazol-4-carboxylic acid, 5-{1-[(adamantan-1-yl)methyl]-5-methyl-1H-pyrazol-4-yl}-2-({6-[(1,3-benzothiazol-2-yl)amino]- 5cyclopropylpyridazin-3-yl}(methyl)amino)-1,3-thiazol-4-carboxylic acid, 2-[[6-( 1,3-benzothiazol-2-ylamino)-5 -methylpyridazin-3 -yl] -[5-[2-(diethylamino)ethylamino] -4hydroxypentyl]amino] -5-[3-[ 4-[3-(dimethylamino)prop-1-ynyl]-2-fluorophenoxy]propyl]thiazole-4carboxylic acid, 2-[[6-( 1,3-Benzothiazol-2-ylamino)-5 -methylpyridazin-3 -yl] -[5-[2-(4-methylpiperazin-1 yl)ethylamino]pentyl]amino]-5- [3-[4-[3-(dimethylamino)prop-1-ynyl]-2-fluorophenoxy]propyl]thiazole-4carboxylic acid, 2-[[6-(1,3-benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]-(4-hydroxy-5-piperazin-1ylpentyl)amino]-5-[3-[4-[ 3-(dimethylamino)prop-1-ynyl]-2-fluorophenoxy]propyl]thiazole-4-carboxylic acid, 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)-5-[(28)-3-{4-[3( dimethylamino)prop-1 -in-1 -yl]-2-fluorophenoxy}-2-methylpropyl] - 1,3-thiazol-4-carboxylic acid, 2-[4-aminobutyl-[6-(1,3-benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]amino]-5-[3-[4-[3(dimethylamino)prop-1- inyl]-2-fluorophenoxy]propyl]thiazole-4-carboxylic acid, 2-[5-aminopentyl-[6-(1,3-benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]amino]-5-[3-[4-[3(dimethylamino)prop-1- inyl]-2-fluorophenoxy]propyl]thiazole-4-carboxylic acid, 2-[[6-( 1,3-benzothiazol-2-ylamino)-5 -methylpyridazin-3 -yl] -[3 -(dimethylamino)propyl]amino] -5-[3[2-fluoro-4-[ 3 -(methylamino)prop-1-ynyl]phenoxy]propyl]thiazole-4-carboxylic acid, 2-[[6-( 1,3-benzothiazol-2-ylamino)-5 -methylpyridazin-3 -yl]methylamino]-5-[3-[4-[3(dimethylamino)but-1-ynyl]-2 -fluorophenoxy]propyl]thiazole-4-carboxylic acid, 2-[[6-( 1,3-benzothiazol-2-ylamino)-5 -methylpyridazin-3 -yl] -[4-hydroxy-5(trimethylammonio)pentyl] amino] -5-[3-[4-[ 3 -(dimethylamino)prop-1-ynyl]-2 fluorophenoxy]propyl]thiazole-4-carboxylate, 3-[2-[3-[4-[3-[2-[[6-(1,3-benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]methylamino]-4carboxythiazol-5 -yl] propoxy]-3-fluorophenyl] prop-2-ynylamino] ethyldimethylammonio] propane-1 sulfonate, 2-[[6-(1,3-benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]-[4-hydroxy-5-(4-methylmorpholin-4-yl4-yl)pentyl]amino]- 5-[3-[4-[3-(dimethylamino)prop-1-ynyl]-2-fluorophenoxy]propyl]thiazole-4carboxylate, 2-[3-[4-[3-[2-[[6-(1,3-benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]methylamino]-4carboxythiazol-5-yl]propoxy]- 3-fluorophenyl]prop-2-ynylamino]ethyltrimethylammonium, 2-[[6-(1,3-benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]-[6-(trimethylammonio)hexyl]amino]-5[3-[2-fluoro-4-[ 3-(methylamino)prop-1-ynyl]phenoxy]propyl]thiazole-4-carboxylate, 2-[[6-( 1,3-benzothiazol-2-ylamino)-5 -methylpyridazin-3 -yl] -[5-[2- 293 046336 hydroxyethyl(dimethyl)ammonio] pentyl] amino] -5-[3 -[4-[3 -(dimethylamino)prop-1-inyl]-2fluorophenoxy]propyl]thiazole-4-carboxylate, 2-[[6-( 1,3-benzothiazol-2-ylamino)-5 -methylpyridazin-3 -yl] -[5[carboxymethyl(dimethyl)ammonio] pentyl] amino] -5-[3-[4-[ 3 -(dimethylamino)prop-1-ynyl]-2fluorophenoxy]propyl]thiazole-4-carboxylate, 3-[[5-[[6-(1,3-benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]-[4-carboxy-5-[3-[2-fluoro-4-[( E)-3oxoprop-1-enyl]phenoxy]propyl]thiazol-2-yl]amino]-2-hydroxypentyl]dimethylammonio]propane-1 sulfonate, 2-[[6-(1,3-benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]methylamino]-5-[3-[4-[3(dimethylamino)prop-1-ynyl]phenoxy] propyl]thiazole-4-carboxylic acid, 2-[[6-( 1,3-benzothiazol-2-ylamino)-5 -methylpyridazin-3 -yl]methylamino]-5-[3-[4-[1[(dimethylamino)methyl]-3-bicyclo[ 1.1.1] pentanyl] phenoxy] propyl] thiazole-4-carboxylic acid, 2-[[6-(1,3-benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]-(3-phosphonopropyl)amino]-5-[3-[2-fluoro4-[3-(methylamino )prop-1-ynyl]phenoxy]propyl]thiazole-4-carboxylic acid, 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(3-carboxypropyl)amino)-5-(3-{4-[3(dimethylamino) prop-1 -yn-1 -yl]-2-fluorophenoxy}propyl)-1,3-thiazol-4-carboxylic acid, 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(3-phosphonopropyl)amino)-5-(3-{4-[3(dimethylamino) prop-1 -yn-1 -yl]-2-fluorophenoxy}propyl)-1,3-thiazol-4-carboxylic acid, 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(3-sulfopropyl)amino)-5-(3-{4-[3(dimethylamino) prop-1 -yn-1 -yl]-2-fluorophenoxy}propyl)-1,3-thiazol-4-carboxylic acid, 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(4-sulfobutyl)amino)-5-(3-{4-[3(dimethylamino) prop-1 -yn-1 -yl]-2-fluorophenoxy}propyl)-1,3-thiazol-4-carboxylic acid, 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(4-phosphonobutyl)amino)-5-(3-{4-[3(dimethylamino) prop-1 -yn-1 -yl]-2-fluorophenoxy}propyl)-1,3-thiazol-4-carboxylic acid, 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(4-carboxybutyl)amino)-5-(3-{4-[3(dimethylamino) prop-1 -yn-1 -yl]-2-fluorophenoxy}propyl)-1,3-thiazol-4-carboxylic acid, 2-[{6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(pent-4-yn-1-yl)amino]-5-(3-{2 -fluoro4-[3 -(methylamino)prop-1 -yn-1 -yl]phenoxy}propyl)-1,3-thiazol-4-carboxylic acid, 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)-5-{3-[2-fluoro-4-(3{ methyl[2-(methylamino)ethyl]amino}prop-1-yn-1-yl)phenoxy]propyl}-1,3-thiazole-4-carboxylic acid, 2-({6-[(1,3-benzothiazole -2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)-5-[3-(2-fluoro-4-{3-[(2hydroxyethyl)(methyl)amino]prop-1 - in-1 -yl} phenoxy)propyl] - 1,3-thiazol-4-carboxylic acid, 2-({6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)-5-{3-[2-fluoro-4-(3hydroxyprop- 1 -yn-1 -yl)phenoxy]propyl}-1,3-thiazol-4-carboxylic acid, 2-[5-azidopentyl-[6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl]amino]-5-[3-[2-fluoro-4[3 - (methylamino)prop-1-ynyl]phenoxy]propyl]-1,3-thiazole-4-carboxylic acid, its enantiomers and diastereoisomers, and their addition salts with a pharmaceutically acceptable acid or base. 30. A compound according to claim 1, selected from the following group of compounds: 6-[{6-[(1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino]-3-[1-({3-[2(dimethylamino)ethoxy ]-5,7-dimethyladamantan-1-yl}methyl)-5-methyl-1H-pyrazol-4-yl]pyridin-2carboxylic acid, 6-[[6-( 1,3-benzothiazol-2-ylamino)-5 -methylpyridazin-3 -yl]methylamino] -3-[1-[[3-[2-(3hydroxypropylamino)ethoxy] -5.7 -dimethyl-1-adamantyl]methyl]-5-methylpyrazol-4-yl]pyridine-2carboxylic acid, 6-[[6-( 1,3-benzothiazol-2-ylamino)-5 -methylpyridazin-3 -yl]methylamino] -3-[1-[[3-[2-(3methoxypropylamino)ethoxy] -5.7 -dimethyl-1-adamantyl]methyl]-5-methylpyrazol-4-yl]pyridin-2 carboxylic acid, 6-[{6-[( 1,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino] -3 -(1 -{[3 -(2-{ [( 3 S)-3,4dihydroxybutyl]amino}ethoxy)-5,7-dimethyladamantan-1-yl]methyl}-5-methyl-1H-pyrazol-4-yl)pyridine2-carboxylic acid, 6-[[6-(1,3-Benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]methylamino]-3-[1-[[3,5-dimethyl-7-(2morpholinoethoxy)-1- adamantyl]methyl]-5-methylpyrazol-4-yl]pyridine-2-carboxylic acid, 6-[[6-( 1,3-benzothiazol-2-ylamino)-5 -methylpyridazin-3 -yl]methylamino] -3-[1-[[3-[2-(2carboxyethylamino)ethoxy] -5.7 -dimethyl-1-adamantyl]methyl]-5-methylpyrazol-4-yl]pyridine-2carboxylic acid, 6-[[6-(1,3-benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]methylamino]-3-[1-[[3-[2-[(3hydroxyphenyl)methylamino]ethoxy] - 5,7-dimethyl-1-adamantyl]methyl]-5-methylpyrazol-4-yl]pyridin-2 carboxylic acid, 2-[[6-( 1,3-benzothiazol-2-ylamino)-5 -methylpyridazin-3 -yl]methylamino]-5-[3-[4-[3(dimethylamino)prop-1-ynyl]-2 -fluorophenoxy]propyl]thiazole-4-carboxylic acid, 2-[[6-(1,3-benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]-(3-hydroxypropyl)amino]-5-[3-[2 - 294 046336 fluoro-4-[3 -(methylamino)prop-1-inyl]phenoxy]propyl]thiazole-4-carboxylic acid, 2-[[6-( 1,3-benzothiazol-2-ylamino)-5 -methylpyridazin-3 -yl]methylamino]-5-[3-[2-fluoro-4-(3 pyrrolidin-1 -ylprop-1 -inyl)phenoxy]propyl]thiazole-4-carboxylic acid, its enantiomers and diastereoisomers, and their addition salts with a pharmaceutically acceptable acid or base. 31. A pharmaceutical composition containing a compound according to any one of claims 1 to 30 or its addition salt with a pharmaceutically acceptable acid or base, in combination with one or more pharmaceutically acceptable excipients. 32. Use of the pharmaceutical composition according to claim 31 as pro-apoptotic agents. 33. Use of the pharmaceutical composition according to claim 31 for the treatment of malignant neoplasms, autoimmune diseases or diseases of the immune system. 34. Use of a pharmaceutical composition according to claim 33, wherein the malignancy is a hematological malignancy or a solid tumor. 35. Use of a pharmaceutical composition according to claim 34, wherein the hematological malignancy is myeloma, especially multiple myeloma, lymphoma, especially non-Hodgkin's lymphoma (NHL), or leukemia, especially chronic lymphocytic leukemia (CLL), T-cell acute lymphoblastic leukemia (T-ALL), B-cell acute lymphoblastic leukemia (BALL) and acute myelocytic leukemia (AML). 36. Use of a pharmaceutical composition according to claim 34, wherein the solid tumor is selected from bladder, brain, breast, uterine, esophageal and liver cancer, colorectal cancer, kidney cancer, melanoma, ovarian cancer, prostate cancer, pancreatic cancer and lung cancer. 37. Use of a compound according to any one of claims 1 to 30 or a salt thereof adducted with a pharmaceutically acceptable acid or base for the treatment of a malignancy selected from myeloma, in particular multiple myeloma, lymphoma, in particular non-Hodgkin's lymphoma (NHL), or leukemia, in features of chronic lymphocytic leukemia (CLL), T-cell acute lymphoblastic leukemia (T-ALL), B-cell acute lymphoblastic leukemia (B-ALL) and acute myelocytic leukemia (AML), cancer of the bladder, brain, breast, uterus , esophagus and liver, colorectal cancer, kidney cancer, melanoma, ovarian cancer, prostate cancer, pancreatic cancer and lung cancer, especially non-small cell lung cancer and small cell lung cancer. 38. Pharmaceutical combination of a compound according to any one of claims 1 to 30 with an antitumor agent selected from genotoxic agents, mitotic poisons, antimetabolites, proteasome inhibitors, kinase inhibitors and antibodies. 39. A pharmaceutical composition containing the combination according to claim 38 in combination with one or more pharmaceutically acceptable excipients. 40. Use of a pharmaceutical combination according to claim 38 for the treatment of malignant neoplasms. 41. Use of a compound according to any one of claims 1 to 30 for the treatment of malignant neoplasms requiring radiotherapy. 42. Use of the pharmaceutical composition according to claim 31 for the treatment of diseases or conditions characterized by excessive or deregulated platelet activity, in particular prothrombotic conditions. 43. A synthesis intermediate that is: *) _where R4 and m are as defined for general formula (I), preferably R4 represents a hydrogen atom.