OA20487A - 3,6-diamino-pyridazin-3-yl derivatives, pharmaceutical compositions containing them and their uses as pro-apoptotic agents. - Google Patents

3,6-diamino-pyridazin-3-yl derivatives, pharmaceutical compositions containing them and their uses as pro-apoptotic agents. Download PDF

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OA20487A
OA20487A OA1202200024 OA20487A OA 20487 A OA20487 A OA 20487A OA 1202200024 OA1202200024 OA 1202200024 OA 20487 A OA20487 A OA 20487A
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methyl
amino
propyl
mmol
benzothiazol
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OA1202200024
Inventor
Maïa CHANRION
James Edward Paul Davidson
James Brooke MURRAY
Andras Kotschy
Attila Paczal
Jérôme-Benoît STARCK
Frédéric COLLAND
Patrice Desos
Tibor Novak
Simon Bedford
Mark Philip DODSWORTH
Petra Dunkel
Andras Herner
Zoltan Madarasz
Ana Leticia MARAGNO
Mark Molnar
Miklos Nyerges
Rachel Jane PARSONS
Monika Rudasova
Agnes Strofek
Marianna SZIGETI
Paul Webb
Matyas Pal Timari
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Les Laboratoires Servier
Vernalis (R&D) Limited
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φ 3,6-DIAMINO-PYRIDAZIN-3-YL DERIVATIVES, PHARMACEUTICAL COMPOSITIONS CONTAINING THEM AND THEIR USES AS PRO-APOPTOTIC AGENTS
FIELD OF THE INVENTION
The présent invention relates to 3,6-diamino-pyridazin-3-yl dérivatives, to pharmaceutical compositions containing them and their uses as pro-apoptotic agents. The compounds of the présent invention inhibit the activity of the Bcl-xL protein and may be of interest in the 5 treatment of cancer, immune and autoimmune diseases.
BACKGROUND OF THE INVENTION
Apoptosis (programmed cell death) is an evolutionarily conserved pathway essential for tissue homeostasis, development and removal of damaged cells. Deregulation of apoptosis contributes to human diseases, including malignancies, neurodegenerative disorders, diseases of the immune system and autoimmune diseases (Hanahan and Weinberg, Cell. 2011 Mar 4;144(5):646-74; Marsden and Strasser, Annu Rev Immunol. 2003;21:71-105; Vaux and Flavell, Curr Opin Immunol. 2000 Dec; 12(6):719-24). Evasion of apoptosis is recognized as a hallmark of cancer, participating in the development as well as the sustained expansion of tumors and the résistance to anti-cancer treatments (Hanahan and Weinberg, Cell. 2000 Jan
7;100(l):57-70).
The Bcl-2 protein family comprises key regulators of cell survival which can suppress (e.g., Bcl-2, Bcl-xL, Mcl-1) or promote (e.g., 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(l):47-59).
In the face of stress stimuli, whether a cell survives or undergoes apoptosis is dépendent on the extent of pairing between the Bcl-2 family members that promote cell death with family members that promote cell survival. For the most part, these interactions involve the docking of the Bcl-2 homology 3 (BH3) domain of proapoptotic family members into a groove on the surface of pro-survival members. The presence of Bcl-2 homology (BH) domain defines the membership of the Bcl-2 family, which is divided into three main groups depending upon the particular BH domains présent within the protein. The prosurvival members such as Bcl-2, BclxL, and Mcl-1 contain BH domains 1^1, whereas Bax and Bak, the proapoptotic effectors of mitochondrial outer membrane permeabilization during apoptosis, contain BH domains 1—3 (Youle and Strasser, Nat. Rev. Mol. Cell Biol. 2008 Jan;9(l):47-59).
Overexpression of the prosurvival members of the Bcl-2 family is a hallmark of cancer and it φ has been shown that these proteins play an important rôle in tumor development, maintenance and résistance to anticancer therapy (Czabotar et al., Nat. Rev. Mol. Cell Biol. 2014 Jan; 15(1):49-63). Bcl-xL (also named BCL2L1, from BCL2-like 1) is frequently amplified in cancer (Beroukhim et aL, Nature 2010 Feb 18;463(7283):899-905) and it has been shown that 5 its expression inversely correlates with sensitivity to more than 120 anti-cancer therapeutic molécules in a représentative panel of cancer cell lines (NCI-60) (Amundson et al., Cancer Res. 2000 Nov l;60(21):6101-10).
In addition, several studies using transgenic knockout mouse models and transgenic overexpression of Bcl-2 family members highlighted the importance of these proteins in the 10 diseases of the immune system and autoimmune diseases (for a review, see Merino et al.,
Apoptosis 2009 Apr; 14(4):570-83. doi: 10.1007/sl0495-008-0308-4.PMID: 19172396).
Transgenic overexpression of Bcl-xL within the T-cell compartment resulted in résistance to apoptosis induced by glucocorticoid, g-radiation and CD3 crosslinking, suggesting that transgenic Bcl-xL overexpression can reduce apoptosis in resting and activated T-cells (Droin 15 et al., Biochim Biophys Acta 2004 Mar l;1644(2-3):179-88. doi:
10.1016/j.bbamcr.2003.10.011.PMID: 14996502 ). In patient samples, persistent or high expression of antiapoptotic Bcl-2 family proteins has been observed (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 rheumatoid arthritis patients exhibited increased Bcl-xL expression 20 and were résistant 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 also shown benefit in pre-clinical models of diseases of the immune system and autoimmune diseases. Treatment with ABT-737 (Bcl-2, Bcl-xL, and Bcl-w inhibitor) resulted in potent inhibition of lymphocyte prolifération in vitro. Importantly, mice treated with ABT- 737 in animal models of arthritis 25 and lupus showed a significant decrease in disease severity (Bardwell et al., J Clin Invest. 1997
Feb l;99(3):439-46. doi: 10.1172/JCU19178.PMID: 9022077). In addition, it has been shown that ABT-737 prevented allogeneic T-cell activation, prolifération, and cytotoxicity in vitro and inhibited 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.l43230 2277.2011.01272.x. Epub 2011 May 25.PMID: 21615547).
The findings indicated above motivated the discovery and development of a new class of drugs named BH3 mimetics. These molécules are able to disrupt the interaction between the proapoptotic and antiapoptotic members of the Bcl-2 family and are potent inducers of apoptosis. This new class of drugs includes inhibitors of Bcl-2, Bcl-xL, Bcl-w and Mcl-1. The first BH3 mimetics described were ABT-737 and ABT-263, targeting Bcl-2, Bcl-xL and Bclw (Park et al., J. Med. Chem. 2008 Nov 13;51(21):6902-15; Roberts et al., J. Clin. Oncol. 2012 5 Feb 10;30(5):488-96). After that, sélective inhibitors of Bcl-2 (ABT-199 and S55746 - Souers et al., Nat Med. 2013 Feb;19(2):202-8; Casara et al., Oncotarget 2018 Apr 13;9(28):2007520088), 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 Transi 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. 10 2015 Jan 15;6:el590.; 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) were also discovered. The sélective Bcl-2 inhibitor ABT-199 is now approved for the treatment of patients with CLL and AML in combination therapy, while the other inhibitors are still under 15 pre-clinical or clinical development. In pre-clinical models, ABT-263 has shown activity in several hematological malignancies and solid tumors (Shoemaker et al., Clin. Cancer
Res. 2008 Jun l;14(ll):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 exhibited objective antitumor activity in lymphoid malignancies 20 (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 investigated in combination with several thérapies in solid tumors. The sélective Bcl-xL inhibitors, A-1155463 or A-1331852, exhibited in vivo activity in pre-clinical models of T-ALL (T-cell Acute Lymphoblastic Leukemia) and different types of solid tumors (Tao et al., ACS Med. Chem. Lett. 2014 Aug 26;5(10):1088-93; Leverson et al., Sci. Transi. Med. 2015 Mar 18;7(279):279ra40). The Mcl-1 sélective inhibitors hâve shown promising in vivo activity in several types of hematological cell malignancies in preclinical models and three of them, S64315, AMG176 and AZD5991, are currently being investigated in clinical trials (Yang et al., Eur. J. Med. Chem. 2019 May 8;177:63-75). Therefore, BH3 mimetics represent a highly attractive approach for the development of novel thérapies in oncology and in the field of immune and autoimmune diseases. In particular, the need exists for small molécules that inhibit selectively the Bcl-xL protein. The présent invention fulfills this need.
SUMMARY OF THE INVENTION
The présent invention provides potent sélective Bcl-xL inhibitors of formula (I) as defined below. We hâve shown that these compounds are able to induce apoptosis of cancer cells in vivo, triggering tumor régression in mice. Based on their pro-apoptotic properties, the compounds of the invention could be of interest for the treatment of pathologies involving a deregulation in apoptosis, such as, for example, cancer, auto-immune diseases and diseases of the immune system. In addition, these compounds were well tolerated in mice, with no clinically relevant body weight loss upon treatment with efficacious doses, indicating a possible therapeutic margin for the use of these Bcl-xL-targeting small molécules in cancer treatment. In agreement with the previously described rôle of Bcl-xL in the régulation of platelets lifespan (Zhang et al., Cell Death Differ. 2007 May;14(5):943-51; Mason et al., Cell. 2007 Mar 23; 128(6): 1173-86), we observed a réduction in the number of circulating platelets after treatment of mice with these inhibitors, with recovery after treatment discontinuation. Considering this effect in platelet survival, the Bcl-xL inhibitors of the présent invention could also be used for treating diseases or conditions characterized by an excess or a deregulated activity of platelets, such as, for example, pro-thrombotic conditions.
DETAILED DESCRIPTION OF THE INVENTION
In a first embodiment (El), the présent invention provides compounds of formula (I):
wherein:
♦ Ri and R2 independently of one another represent a group selected from: hydrogen; linear or branched Ci-Côalkyl optionally substituted by a hydroxyl or a Ci-Côalkoxy group; Cs-Cecycloalkyl; trifluoromethyl; linear or branched Ci-Côalkylene-heterocycloalkyl wherein the heterocycloalkyl group is optionally substituted by a a linear or branched Ci-Côalkyl group;
or Ri and R2 form with the carbon atoms carrying them a Cs-Côcycloalkylene group,
R3 represents a group selected from: hydrogen; C3-C6cycloalkyl; linear or branched CiC6alkyl; -Xi-NRaRb; -Xi-N+RaRbRc; -Xi-O-Rc; -Xi-COORc; -Xi-PO(OH)2; -XiSO2(OH); -X1-N3 and :
— X1-=CH ♦ Ra and Rb independently of one another represent a group selected from: hydrogen; heterocycloalkyl; -SO2-phenyl wherein the phenyl may be substituted by a linear or branched Ci-Côalkyl; linear or branched Ci-Côalkyl optionally substituted by one or two hydroxyl groups; Ci-C6alkylene-SO2OH; Ci-Côalkylene-SO2O'; Ci-CôalkyleneCOOH; Ci-C6alkylene-PO(OH)2; Ci-C6alkylene-NRdRe; Ci-C6alkylene-N+RdReRf; CiCealkylene-phenyl wherein the phenyl may be substituted by a Ci-Côalkoxy group;
the group:
or Ra and Rb form with the nitrogen atom carrying thema cycle Bi;
or Ra, Rb and Rc form with the nitrogen atom carrying thema bridged C3-Csheterocycloalkyl, ♦ Rc, Rd, Re, Rr, independently of one another represents a hydrogen or a linear or branched Ci-Cealkyl group, or Rd and Re form with the nitrogen atom carrying thema a cycle B2, or Rd, Re and Rf form with the nitrogen atom carrying thema bridged C3-Csheterocycloalkyl, ♦ Heti represents a group selected from:
♦ Ai is -NH-, -N(Ci-C3aikyl), O, S or Se, ♦ A2 is 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)2RG3, -S(O)2NRGiRG2,
-NRGiS(O)2Rg2, -NRgiC(=NRg2)NRgiRg2, -C(=S)NRgiRg2, -C(=NRGi)NRGiRg2, halogen, -NO2, and -CN, in which:
- Rgi and Rg2 at each occurrence are each independently selected from the group consisting of hydrogen, Ci-Côalkyl optionally substituted by 1 to 3 halogen atoms, 5 C2-C&alkenyl, Cz-Cealkynyl, Cs-Côcycloalkyl, phenyl and -(CH2)i-4-phenyl;
- Rgs is selected from the group consisting of Ci-Côalkyl optionally substituted by 1 to 3 halogen atoms, C2-Côalkenyl, C2-Côalkynyl, Cs-Côcycloalkyl, phenyl and -(CH2)i-4-phenyl;
or
Rgi and Rg2, together with the atom to which each is attached are combined to form a C310 Csheterocycloalkyl ; or in the alternative, G is selected from the group consisting of:
wherein Rg4 is selected from Ci-Cealkyl optionally substituted by 1 to 3 halogen atoms, C2-C6alkenyl, C2-Côalkynyl and Cs-Cecycloalkyl, ♦ R4 represents a hydrogen, fluorine, chlorine or bromine atom, a methyl, a hydroxyl or a methoxy group, ♦ R5 represents a group selected from: Ci-Côalkyl optionally substituted by 1 to 3 halogen atoms; C2-Côalkenyl; C2-C6alkynyl; halogen or -CN, ♦ Rô represents a group selected from:
hydrogen;
-Cz-Cealkenyl;
-X2-O-R7;
-X2-NSO2-R7;
-C=C(R9)-Yi-O-R7;
Cs-Cecycloalkyl;
Cs-Côheterocycloalkyl optionally substituted by a hydroxyl group;
C3-C6cycloalkylene-Y2-R7 ;
C3-C6heterocycloalkylene-Y2-R7 group, an heteroarylene-R? group optionally substituted by a linear or branched Ci-Côalkyl group, ♦ R7 represents a group selected from: linear or branched Ci-Cealkyl group; (Cs-Côjcycloalkylene-Rs; or:
wherein Cy represents a C3-C8cycloalkyl, ♦ Rs represents a group selected from: hydrogen; linear or branched Ci-Cealkyl, 5 -NR’aR’b; -NR’a-C0-0R’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’aR’b, -X’2-NR’aR’b,-NR’c-X’2-N3 and :
-NR'C—X'2^CH ?
♦ R9 represents a group selected from linear or branched Ci-Côalkyl, trifluoromethyl, hydroxyl, halogen, Ci-Cealkoxy, ♦ Rio represents a group selected from hydrogen, fluorine, chlorine, bromine, -CF3 and methyl, ♦ Ru represents a group selected from hydrogen, Ci-C3alkylene-R8, -O-Ci-C3alkyleneRs, -CO-NRhRi and -CH=CH-Ci-C4alkylene-NRhRi, -CH=CH-CHO, C3Cscycloalkylene-CH2-R8, C3-C8heterocycloalkylene-CH2-R8, ♦ R12 and R13, independently of one another, represent a hydrogen atom or a methyl group, ♦ Ri4 and R15, independently of one another, represent a hydrogen or a methyl group, or Ri4 and R15 form with the carbon atom carrying them a a cyclohexyl, ♦ Rh and Ri, independently of one another, represent a hydrogen or a linear or branched 20 Ci-Cûalkyl group, ♦ Xi and X2 independently of one another, represent a linear or branched Ci-Côalkylene group optionally substituted by one or two groups selected from trifluoromethyl, hydroxyl, halogen, Ci-Côalkoxy, ♦ X’2 represents a linear or branched Ci-Cealkylene, ♦ R’a and R’b independently of one another, represent a group selected from: hydrogen;
heterocycloalkyl; -SO2-phenyl wherein the phenyl may be substituted by a linear or branched Ci-Côalkyl; linear or branched Ci-Cealkyl optionally substituted by one or two hydroxyl or Ci-C6alkoxy groups; Ci-Câalkylene-SO2OH; Ci-C6alkylene-SO2O';
Ci-C6alkylene-COOH; Ci-C6alkylene-PO(OH)2; Ci-C6alkylene-NR’dR’e; Ci-C6alkylene-N+R’dR’eR’f; Ci-Coalkylene-O-Ci-Coalkylene-OH; Ci-Csalkylenephenyl wherein the phenyl may be substituted by a hydroxyl or a Ci-Côalkoxy group; the group:
or R’a and R’b form with the nitrogen atom carrying them a cycle B3, or R’ a, R’b and R’c form with the nitrogen atom carrying them a bridged Cs-Csheterocycloalkyl, ♦ R’c, R’d, R’e, R’f, independently of one another, represents a hydrogen or a linear or branched Ci-Côalkyl group, or R’d and R’e form with the nitrogen atom carrying them a cycle B4, or R’d, R’e and R’f form with the nitrogen atom carrying them a bridged C3-Csheterocycloalkyl, ♦ Yi represents a linear or branched Ci-C4alkylene, ♦ Y2 represents a bond, -O-, -O-CH2-, -O-CO-, -O-SO2-, -CH2-, -CH2-O, -CH2-CO-, -CH2-SO2-,-C2H5-, -CO-, -CO-O-, -CO-CH2-, -CO-NH-CH2-, -SO2-, -SO2-CH2-, -NH-CO-, -NH-SO2-, ♦ m=0,1 or 2, ♦ p=l, 2, 3 or 4, ♦ Bi, B2, B3 and B4, independently of one another, represents a Cs-Csheterocycloalkyl group, which group can: (i) be a mono- or bi-cyclic group, wherein bicyclic group includes fused, briged or spiro ring system, (ii) can contain, in addition to the nitrogen atom, one or two hetero atoms selected independently from oxygen, sulphur and nitrogen, (iii) be substituted by one or two groups selected from: fluorine, bromine, chlorine, linear or branched Ci-Côalkyl, hydroxyl, -NH2, oxo or piperidinyl, it also being understood that:
aryl means a phenyl, naphthyl, biphenyl or indenyl group, heteroaryl means any mono- or bi-cyclic group composed of from 5 to 10 ring members, having at least one aromatic moiety and containing from 1 to 4 hetero atoms selected from oxygen, sulphur and nitrogen (including quaternary nitrogens), cycloalkyl means any mono- or bi-cyclic 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 bi-cyclic non-aromatic carbocyclic group, composed of from 3 to 10 ring members, and containing from one to 3 hetero atoms selected from oxygen, sulphur, SO, SO2 and nitrogen, it being understood that bicyclic group may be fused or spiro type, heteroarylene, cycloalkylene, heterocycloalkylene mean a divalent heteroaryl, cycloalkyl and heterocycloalkyl, its enantiomers and diastereoisomers, and addition salts thereof with a pharmaceutically acceptable acid or base.
Among the pharmaceutically acceptable acids there may be mentioned, without implying any limitation, hydrochloric acid, hydrobromic acid, sulphuric acid, phosphonic acid, acetic acid, trifluoroacetic acid, lactic acid, pyruvic acid, malonic acid, succinic acid, glutaric acid, fumaric acid, tartaric acid, maleic acid, citric acid, ascorbic acid, oxalic acid, methanesulphonic acid and camphoric acid.
Among the pharmaceutically acceptable bases there may be mentioned, without implying any limitation, sodium hydroxide, potassium hydroxide, triethylamine and tert-butylamine.
Further enumerated embodiments (E) of the invention are described herein. It will be recognized that features specified in each embodiment may be combined with other specified features to provide further embodiments of the présent invention.
E2. Compound according to El wherein Heti represents:
or
E3. Compound according to El or E2 wherein Ri represents a methyl group and R2 represents a hydrogen atom.
E4. Compound according to El or E2 wherein Ri and R2 represent each a methyl group or a hydrogen atom.
E5, Compound according to El or E2 wherein Ri or R2 represents a linear or branched CiCc, alkyl optionally substituted by a hydroxyl or a Ci-Cealkoxy group, preferably a group selected from: methyl, methoxymethyl, hydroxymethyl, ethyl and prop-2-yl.
E6, Compound according to El or E2 wherein Ri or R2 represents a Cs-Côcycloalkyl, preferably a cyclopropyl group.
E7. Compound according to El or E2 wherein Ri represents a trifluoromethyl group.
E8, Compound according to El or E2 wherein R2 represents a linear or branched Ci-Cealkylene-heterocycloalkyl, preferably a (4-methyl-piperazin-l-yl)propyl group.
E9. Compound according to El or E2 wherein Ri and R2 form with the carbon atoms carrying them a cyclohexene or a cyclopentene group.
E10, Compound according to any of El to E9 wherein R3 represents a hydrogen atom or a methyl group.
Eli, Compound according to any of El to E9 wherein R3 represents -Xi-PO(OH)2, -Xi20 SO2(OH), -Xi-NRaRt>; -Xi-N+RaRbRc, wherein Ra or Rb, or both of them, represent a group selected from Ci-C6alkylene-SO2OH, Ci-C6alkylcne-SO2O’ and Ci-C6alkylene-PO(OH)2.
E12. Compound according to any of El to E9 wherein R8 represents -NR’aR’b; -N+R’aR’bR’c; -NH-X’2-N+R’aR’bR’c, wherein R’a and R’b, or both of them, represent a group selected from Ci-C6alkylene-SO2OH and Ci-C6alkylene-PO(OH)2.
E13. Compound according to any of El to E9 wherein R3 represents a group selected from: ethyl, propyl; 2-methoxy-ethyl, 2-(morpholin-4-yl)ethyl, 3-(morpholin-4-yl)propyl, 3diethylamino-propyl, 3-methoxy-propyl, 3-hydroxy-propyl, 3,4-dihydroxy-butyl, 4-methoxy3-hydroxy-butyl, 4-hydroxy-3-methoxy-butyl, 2,3-dihydroxy-propyl, 4,5-dihydroxy-pentyl, 4hydroxy-butyl, 3-hydroxy-2-morpholino-propyl, 5-hydroxy-4-methoxy-pentyl, 5-morpholino4-hydroxy-pentyl, 3-hydroxy-2-methoxy-propyl, 5-[2-(dimethylamino)ethyl-methyl-amino]4-hydroxy-pentyl, 5-hydroxy-pentyl, 5-methoxy-4-hydroxy-pentyl, 5-(dimethylamino)-4hydroxy-pentyl, 4-hydroxy-5-(trimethylammonio)pentyl, 5-[3-sulfonate-propyl-dimethylammonio]-4-hydroxy-pentyl, 4-hydroxy-5-(methylamino)pentyl, 3-carboxy-propyl, 5[methyl(4-piperidyl)amino]pentyl, 5-(2-morpholinoethylamino)pentyl, 5-[2-(4-methylpiperazin-l-yl)ethylamino)pentyl, 4-[2-(4-methyl-piperazin-l-yl)ethylamino)butyl, 4hy droxy-5 - [methy 1- [2-(methylamino)ethyl] amino]pentyl, 5 - [2-(diethylamino)ethy lamino] -4hydroxy-pentyl, 5-(4-Amino-l-piperidyl)-4-hydroxy-pentyl, 4-hydroxy-5-piperazin-l-ylpentyl, 5-[2-(l-piperidyl)ethylamino]pentyl, 4-(4-Amino-l-piperidyI)butyl, 4-[2(diethy lamino)ethylamino]butyl, 5 -(4-Amino-1 -piperidy l)pentyl, 4-[methyl- [2(methylamino)ethyl]amino]butyl, 4-(2-morpholinoethylamino)butyl, 4-[2-(4-methylpiperazin-1 -y l)ethylamino)butyl, 4-[(l -methy l-4-piperidyl)amino]butyl, 5 - [2(diethylamino)ethylamino]pentyl, 4-piperazin-l-ylbutyl, 4-(methylamino)butyl, 5-piperazin-lylpentyl, 5-[methyl-[2-(methylamino)ethyl, 5-aminopentyl, 4-aminobutyl, 5(methylamino)pentyl, 5-[3-(4-methylpiperazin-l-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-sulfonate-propyl-dimethyl-ammonio]pentyl, 4(trimethylammonio)buty 1, 4-hy droxy-5-quinuclidin-1 -ium-1 -yl-pentyl, 4-hy droxy-5-(1 -aza-4azoniabicyclo[2.2.2]octan-4-yl)pentyl, 4-hydroxy-5-(4-methylmorpholin-4-ium-4-yl)pentyl, 5-(trimethylammonio)penty 1, 4-hy droxy-5-(l -methy Ipiperidin-1 -ium-1 -yl)penty 1, 5-(1,4dimethylpiperazin-1 -ium-1 -y l)-4-hy droxy-pentyl, 6-(trimethy lammonio)hexy 1, 5 - [3 hydroxypropyl(dimethyl)ammonio]pentyl, 5-[2-hydroxyethyl(dimethyl)ammonio]pentyl, 520487
[carboxymethyl(dimethyl)ammonio]pentyl, 5-[carboxymethyl(dimethyl)ammonio]pentyl, 3phosphonopropyl, 3-[3-sulfonate-propyl-dimethyl-ammonio]propyl, 3-sulfopropyl, 4sulfobutyl, 4-phosphonobutyl, 4-carboxybutyl, 3-aminopropyl, 3-azidopropyl, pent-4-yn-l-yl, 4-aminobutyl, 4-azidobutyl, hex-5-yn-l-yl, 5-azidopentyl.
E14, Compound according to any of El to E13 wherein Het2 represents:
E15, Compound according to any of El to E13 wherein Het2 represents:
COOH
E16. Compound according to E14 or E15 wherein Rô represents a hydrogen atom.
E17. Compound according to E14 wherein Rô represents a -X2-0-R? group wherein X2 is a propylene group.
E18. Compound according to E17 wherein R? represents the following group:
E19. Compound according to E17 wherein R? represents the following group:
E20, Compound according to E17 wherein R? represents the following group:
E2L The compound according to any of E18 to E20 wherein R8 represents NR’aR’b,
E22. Compound according to any of E18 to E20 wherein R8 represents a group selected from: dimethylamino, methylamino, methylethylamino, diethylamino, methyl[2(methylamino)ethyl]amino, (2-hydroxyethyl)(methyl)amino, 4-morpholinyl, pyrrolidin-l-yl, 5 1-piperidyl, [(tert-butoxy)carbonyl](methyl)amino, hydroxyl, bis(3-sulfopropyl)amino, 3sulfopropylamino, methyl(3-sulfopropyl)amino, methyl(p-tolylsulfonyl)amino, (4methoxyphenyl)methyl-methyl-amino, 2-(dimethylamino)ethylamino and :
E23, Compound according to to any of E18 to E20 wherein R8 represents a group selected from: 3-piperazin-l-yl, 4-methylpiperazin-l-yl, 3-hydroxypropyl(methyl)amino, [(35)-3,410 dihydroxybutyl]-methyl-amino, 3-hydroxypropylamino, [(35)-3,4-dihydroxybutyl]amino, 4hydroxybutyl(methyl)amino, [(3R)-3,4-dihydroxybutyl]-methyl-amino, 4-hydroxybutylamino, [(3R)-3,4-dihydroxybutyl]amino.
E24, Compound according to E17 wherein R7 represents:
wherein Ru is selected from dimethylcarbamoyl, 3-(dimethylamino)propyl, 3(methylamino)propyl, 3-(methylamino)propyl.
£25* Compound according to E14 wherein Rô represents a C3-C6heterocycloalkylene-Y2-R7 group wherein the heterocycloalkylene group is selected from:
E26, Compound according to E25 wherein R7 is selected from: methyl, methylethyl, tertbutyl, 2-methylpropyl and phenyl, preferably phenyl.
E27, Compound according to E14 or E15 wherein Rô represents -C=C(R9)-Yi-O-R7 wherein
Yi is a methylene group.
E28. Compound according to E14 or E15 wherein Rô represents:
E29. The compound according to E28 wherein R7 represents a group selected from:
wherein Rsrepresents -O-X’2-NR’aR’b or -X’2-NR’aR’b, preferably -O-X’2-NR’aR’b.
E30, Compound according to E28 wherein R7 represents a group selected from :
wherein Rs represents a group selected from: hydrogen, 2-(methylamino)ethoxy and:
E31, The compound according to E28 wherein R7 represents a group selected from :
wherein Rg represents a group selected from: 2-(dimethylamino)ethoxy, 2-[(25 sulfoethyl)amino]ethoxy, 2-[methyl(2-sulfoethyl)amino]ethoxy,2-(3hydroxypropylamino)ethoxy, 2-(3-methoxypropylamino)ethoxy, 2-morpholinoethoxy, 2-(2carboxyethylamino)ethoxy, 2-[(3-hydroxyphenyl)methylamino]ethoxy,2(methylamino)ethoxy, 2-pyrrolidin-l-ylethoxy, 2-(4-methylpiperazin-l-yl)ethoxy, 2-(4hydroxybutylamino)ethoxy, 2-piperazin-l-ylethoxy,2-[310 hydroxypropyl(methyl)amino]ethoxy, 2-[4-hydroxybutyl(methyl)amino]ethoxy; 2-[[3hydroxy-2-(hydroxymethyl)propyl]amino]ethoxy; 2-[bis(3-hydroxypropyl)amino]ethoxy.
E32, A compound according to El selected in the following group:
2-({6-[(l,3-Benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(4hy droxy buty l)amino)-5-(3- { 2-fluoro-4-[3-(methy lamino)prop-1 -y η-1 yl]phenoxy}propy 1)-1,3 -thiazole-4-carboxylic acid,
2-[[6-(l,3-Benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-methyI-amino]-5-[3-[4[3-(dimethylamino)prop-l-ynyl]-2-fluoro-phenoxy]propyl]thiazole-4-carboxylic acid, 2-({ 6-[(l ,3-Benzothiazol-2-yl)amino] -5 -methy lpyridazin-3 -yl} (methy l)amino)-5 -(3 { 2-fluoro-4- [3-(methylamino)prop-1 -yn-1 -y l]phenoxy }propy 1)-1,3-thiazole-4carboxylic acid, 2-[[6-(l,3-Benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-(3,4dihydroxybutyl)amino]-5-[3-[2-fluoro-4-[3-(methylamino)prop-lynyl]phenoxy]propyl]thiazole-4-carboxylic acid, 2-[[6-(l,3-Benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-(3-hydroxy-4-methoxybutyl)amino] -5 - [3- [2-fluoro-4-[3 -(methylamino)prop-1 -yny l]phenoxy ]propy 1] thiazole4-carboxylic acid, 2-[[6-(l,3-Benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-(2,3dihy droxypropy l)amino] -5 - [3- [2-fluoro-4- [3-(methy lamino)prop-1 ynyl]phenoxy]propyl]thiazole-4-carboxylic acid, 2-[[6-(l,3-Benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-(4,5dihy droxypenty l)amino] -5 - [3- [2-fluoro-4- [3 -(methy lamino)prop-1 ynyl]phenoxy]propyl]thiazole-4-carboxylic acid, 2-[[6-(l,3-Benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-(3,4dihydroxybutyl)amino]-5-[3-[4-[3-(dimethylamino)prop-l-ynyl]-2-fluorophenoxy]propyl]thiazole-4-carboxylic acid, 2-[[6-(l,3-Benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-(3hy droxypropy l)amino] -5 - [3 - [2-fluoro-4- [3-(methy lamino)prop-1 ynyl]phenoxy]propyl]thiazole-4-carboxylic acid, 2-[[6-(l,3-Benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-[4-hydroxy-5(trimethylammonio)pentyl]amino]-5-[3-[2-fluoro-4-[3-(methylamino)prop-lynyl]phenoxy]propyl]thiazole-4-carboxylate,
3-[[5-[[6-(l,3-Benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-[4-carboxy-5-[3-[2fluoro-4-[3-(methylamino)prop-l-ynyl]phenoxy]propyl]thiazol-2-yl]amino]-2hydroxy-pentyl]-dimethyl-ammonio]propane-l-sulfonate, 2-[[6-(l,3-Benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-(4hy droxybuty l)amino] -5- [3 - [2-fluoro-4- [3 - [methyl(3-sulfopropy l)amino]prop-1 ynyl]phenoxy]propyl]thiazole-4-carboxylic acid, 5-{l-[(Adamantan-l-yl)methyl]-5-methyl-l/7-pyrazol-4-yl}-2-({6-[(l,3-benzothiazol
2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)-l,3-thiazole-4-carboxylic acid,
5-{l-[(Adamantan-l-yl)methyl]-5-methyl-lH-pyrazol-4-yl}-2-({6-[(l,3-benzothiazol2-yl)amino]-5-cyclopropylpyridazin-3-yl}(methyl)amino)-l,3-thiazole-4-carboxylic acid,
2-[[6-(l,3-Benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-[5-[2(diethylamino)ethylamino]-4-hydroxy-pentyl]amino]-5-[3-[4-[3(dimethylamino)prop-l-ynyl]-2-fluoro-phenoxy]propyl]thiazole-4-carboxylic acid, 2-[[6-(l,3-Benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-[5-[2-(4methylpiperazin-l-yl)ethylamino]pentyl]amino]-5-[3-[4-[3-(dimethylamino)prop-lynyl]-2-fluoro-phenoxy]propyl]thiazole-4-carboxylic acid, 2-[[6-(l,3-Benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-(4-hydroxy-5-piperazin1 -yl-pentyl)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 -methy Ipy ridazin-3-y 1} (methy l)amino)-5 [(2S')-3-{4-[3-(dimethylamino)prop-l-yn-l-yl]-2-fluorophenoxy}-2-methylpropyl]l,3-thiazole-4-carboxylic acid,
2-[4-Aminobutyl-[6-(l,3-benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]amino]-5[3-[4-[3-(dimethylamino)prop-l-ynyl]-2-fluoro-phenoxy]propyl]thiazole-4-carboxylic acid,
2-[5-Aminopentyl-[6-(l,3-benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]amino]5 - [3 - [4- [3-(dimethy lamino)prop-1 -ynyl] -2-fluoro-phenoxy ]propy 1] thiazole-4carboxylic acid,
2-[[6-(l,3-Benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-[3(dimethy lamino)propy 1] amino] -5 - [3- [2-fluoro-4- [3 -(methy lamino)prop-1 ynyl]phenoxy]propyl]thiazole-4-carboxylic acid,
2-[[6-(l,3-Benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-methyl-amino]-5-[3-[4[3-(dimethylamino)but-1 -ynyl]-2-fluoro-phenoxy ]propyl]thiazole-4-carboxylie acid, 2-[[6-(l,3-Benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-[4-hydroxy-5(trimethylammonio)pentyl]amino]-5-[3-[4-[3-(dimethylamino)prop-l-ynyl]-2-fluorophenoxy]propyl]thiazole-4-carboxylate, 3-[2-[3-[4-[3-[2-[[6-(l,3-Benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-methylamino]-4-carboxy-thiazol-5-yl]propoxy]-3-fluoro-phenyl]prop-2-ynylamino]ethyldimethyl-ammonio]propane-l-sulfonate, 2-[[6-(l,3-Benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-[4-hydroxy-5-(4 methy lmorpholin-4-ium-4-y l)pentyl] amino] -5 - [3- [4- [3 -(dimethy lamino)prop-1 -y nyl] 2-fluoro-phenoxy]propyl]thiazole-4-carboxylate,
2-[3-[4-[3-[2-[[6-(l,3-Benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-methylamino]-4-carboxy-thiazol-5-yl]propoxy]-3-fluoro-phenyl]prop-2-ynylamino]ethyltrimethyl-ammonium,
2-[[6-(l,3-Benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-[6(trimethy lammonio)hexyl] amino] -5 - [3 -[2-fluoro-4- [3-(methylamino)prop-1 ynyl]phenoxy]propyl]thiazole-4-carboxylate,
2- [ [6-(l ,3-Benzothiazol-2-y lamino)-5 -methy 1-py ridazin-3 -y 1] - [5 - [2hy droxy ethy l(dimethy l)ammonio]pentyl] amino] -5 - [3 -[4- [3 -(dimethy lamino)prop-1 ynyl]-2-fluoro-phenoxy]propyl]thiazole-4-carboxylate, 2-[[6-(l,3-Benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-[5[carboxymethy l(dimethy l)ammonio]penty 1] amino] -5 - [3- [4- [3-(dimethylamino)prop-1 ynyl]-2-fluoro-phenoxy]propyl]thiazole-4-carboxylate,
3-[[5-[[6-(l,3-Benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-[4-carboxy-5-[3-[2fluoro-4- [(E)-3 -oxoprop-1 -eny l]phenoxy ]propyl] thiazol-2-y 1] amino] -2-hy droxypentyl]-dimethyl-ammonio]propane-l-sulfonate,
2-[[6-(l,3-Benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-methyl-amino]-5-[3-[4[3-(dimethylamino)prop-l-ynyl]phenoxy]propyl]thiazole-4-carboxylic acid, 2-[[6-(l,3-Benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-methyl-amino]-5-[3-[4[ 1 - [(dimethy lamino)methyl] -3-bicy cio [1.1.1 ]pentanyl]phenoxy ]propy 1] thiazole-4carboxylic acid,
2-[[6-(l,3-Benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-(3phosphonopropyl)amino] -5 - [3- [2-fluoro-4- [3-(methylamino)prop-1 ynyl]phenoxy]propyl]thiazole-4-carboxylic acid,
2-({6-[(l ,3-Benzothiazol-2-y l)amino] -5 -methy Ipy ridazin-3 -yl}(3carboxypropy l)amino)-5-(3- {4- [3-(dimethy lamino)prop- 1-y η-1 -yl]-2fluorophenoxy }propyl)-1,3 -thiazole-4-carboxylic acid,
2-({ 6- [(1,3 -Benzothiazol-2-yl)amino] -5 -methylpy ridazin-3-y 1} (3 phosphonopropy l)amino)-5 -(3 - {4- [3-(dimethylamino)prop-1 -y η-1 -y 1] -2fluorophenoxy}propyl)-l,3-thiazole-4-carboxylic acid,
- 2-({6-[(l,3-Benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(3-sulfopropyl)amino)5-(3-{4-[3-(dimethylamino)prop-l-yn-l-yl]-2-fluorophenoxy}propyl)-l,3-thiazoIe-4carboxylic acid,
2-({6-[(l,3-BcnzothiazoI-2-yl)amino]-5-methylpyridazin-3-yl}(4-sulf'obutyl)amino)5-(3-{4-[3-(dimethylamino)prop-l-yn-l-yl]-2-fluorophenoxy}propyl)-l,3-thiazole-4carboxylic acid, 2-({6-[(l,3-Benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(4phosphonobuty l)amino)-5 -(3 -{4- [3-(dimethylamino)prop-1 -yn-1 -y 1] -2fluorophenoxy }propyl)-1,3 -thiazole-4-carboxylie acid, 2-({6-[(l,3-Benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(4carboxybutyl)amino)-5-(3-{4-[3-(dimethylamino)prop-l-yn-l-yl]-2fluorophenoxy}propyl)-l,3-thiazole-4-carboxylic acid, 2-[{6-[(l,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(pent-4-yn-l-yl)amino]5-(3-{2-fluoro-4-[3-(methylamino)prop-l-yn-l-yl]phenoxy}propyl)-l,3-thiazole-4carboxylic acid, 2-({6-[(l,3-Benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)-5-{3[2-fluoro-4-(3-{methyl[2-(methylamino)ethyl]amino}prop-l-yn-lyl)phenoxy]propyl}-l,3-thiazole-4-carboxylic acid, 2-({6-[(l,3-Benzothiazol-2-yl)amino] -5 -methylpy ridazin-3 -y 1} (methyl)amino)-5 -[3 (2-fluoro-4-{3-[(2-hydroxyethyl)(methyl)amino]prop-l-yn-l-yl}phenoxy)propyl]-l,3thiazole-4-carboxylic acid, 2-({6-[(l,3-Benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)-5-{3[2-fluoro-4-(3-hydroxyprop-l-yn-l-yl)phenoxy]propyl}-l,3-thiazole-4-carboxylic acid, 2-[5-azidopentyl-[6-[(l,3-benzothiazol-2-yl)amino]-5-methyl-pyridazin-3-yl]amino]5 - [3- [2-fluoro-4- [3-(methy lamino)prop-1 -ynyl]phenoxy ]propyl] -1,3 -thiazole-4carboxylic acid, its enantiomers and diastereoisomers, and addition salts thereof with a pharmaceutically acceptable acid or base.
E33. A compound according to El selected in the following group:
6-[{6-[(l,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino]-3-[l({3- [2-(dimethy lamino)ethoxy ] -5,7-dimethy ladamantan-1 -yl }methy l)-5 -methy 1- 1/fpyrazol-4-yl]pyridine-2-carboxylic acid, 6-[[6-(l,3-benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-methyl-amino]-3-[l-[[3[2-(3-hydroxypropylamino)ethoxy]-5,7-dimethyl-l-adamantyl]methyl]-5-methylpyrazol-4-yl]pyridine-2-carboxylic acid,
6-[[6-(l,3-benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-methyl-amino]-3-[l-[[3[2-(3 -methoxypropylamino)ethoxy] -5,7-dimethy 1-1 -adamantyl]methy 1] -5 -methy 1pyrazol-4-yl]pyndine-2-carboxylic acid,
6-[{6-[(l,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino]-3-(l{[3-(2-{ [(35)-3,4-dihydroxybutyl]amino}ethoxy)-5,7-dimethyladamantan-lyl]methyl}-5-methyl-l/7-pyrazol-4-yl)pyridine-2-carboxylic acid,
6-[[6-(l,3-benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-methyl-amino]-3-[l[[3,5-dimethyl-7-(2-morpholinoethoxy)-l-adamantyl]methyl]-5-methyl-pyrazol-4yl]pyridine-2-carboxylic acid,
6-[[6-(l,3-benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-methyl-amino]-3-[l-[[3[2-(2-carboxy ethylamino)ethoxy] -5,7-dimethy 1-1 -adamanty l]methyl] -5 -methy 1pyrazol-4-yl]pyridine-2-carboxylic acid,
6-[[6-(l,3-benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-methyl-amino]-3-[l-[[3[2-[(3-hydroxyphenyl)methylamino]ethoxy]-5,7-dimethyl-l-adamantyl]methyl]-5methyl-pyrazol-4-yl]pyridine-2-carboxylic acid,
2-[[6-(l,3-benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-methyl-amino]-5-[3-[4[3-(dimethylamino)prop-l-ynyl]-2-fluoro-phenoxy]propyl]thiazole-4-carboxylic acid, 2-[[6-(l,3-benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-(3hy droxypropy l)amino] -5 - [3- [2-fluoro-4- [3 -(methy lamino)prop-1 ynyl]phenoxy]propyl] thiazole-4-carboxylic acid,
- 2-[[6-(l,3-benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-methyl-amino]-5-[3-[2fluoro-4-(3-pyrrolidin-l-ylprop-l-ynyl)phenoxy]propyl]thiazole-4-carboxylic acid, its enantiomers and diastereoisomers, and addition salts thereof with a pharmaceutically acceptable acid or base.
E34, Process for the préparation of a compound of formula (I) according to any of El to E33, which process is characterized in that there is used as starting material the compound of formula (II):
wherein Ri and R2 are as defined in formula (I), which 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), of a base (preferably CS2CO3 and DIPEA), of a phosphine (preferably Xantphos) and of the compound of formula (ΠΙ):
wherein R4 and m are as defined in formula (I), to yield the compound of formula (IV):
(IV) which amino function of compound of formula (IV) is further protected to yield the compound of formula (V):
(V) wherein ( P.Gy represents a protecting group (preferably a 2-trimethylsilylethoxymethyl group), which compound of formula (V) is further subjected to a Buchwald reaction, in an aqueous or organic medium, in the presence of a palladium catalyst (preferably Pd2(dba)3), of a base (preferably Cs2CO3 and DIPEA), of a phosphine (preferably Xantphos) and of the compound of formula (VI):
wherein Ai, R3 and Rô are as defined in formula (I) and Alk represents a Ci-Côalkyl group, to yield the compound of formula (Vll-a) or (VII-b):
the benzothiazole group of which compound of formula (Vll-a) or (VII-b) is deprotected (using 5 preferably HFxPyr) and the ester function is hydrolysed (using preferably LiOHxHîO) to yield the compound of formula (I), which compound of formula (I) may be purified according to a conventional séparation technique, which may be converted into its addition salts with a pharmaceutically acceptable acid or base and which is optionally separated into its isomers according to a conventional 10 séparation technique, it being understood that, at any time considered appropriate in the course of the above-described process, hydroxy, amino, carboxylic and phosphono groups of the reagents or intermediates of synthesis may be protected and then deprotected according to the requirements of synthesis.
E35. Process according to E34 wherein the compound (Vl-b) is:
and wherein R7 is selected from the group:
wherein Rs, R12 and R13 are as defined in formula (I).
E36, Process for the préparation of a compound of formula (I) according to E28 wherein Re represents:
and R7 is as defined in formula (I), which process is characterized in that there is used as starting material the compound of formula (II):
(Π) wherein Ri and R2 are as defined in formula (I), which 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), of a base (preferably Cs2CO3 and DIPEA), of a phosphine (preferably Xantphos) and of the compound of formula (III):
Cl
(HD wherein R4 and m are as defined in formula (I), to yield the compound of formula (IV):
(IV) which amino function of compound of formula (IV) is further protected to yield the compound of formula (V):
wherein group),
(V) represents a protecting group (preferably a 2-trimethylsilylethoxymethyl which compound of formula (V) is further subjected to a Buchwald reaction, in an aqueous or organic medium, in the presence of a palladium catalyst (preferably Pd2(dba)3), of a base (preferably CS2CO3 and DIPEA), of a phosphine (preferably Xantphos) and of the compound of formula (VI’):
(VI1) wherein Ai and R3 are as defined in formula (I) and Alk represents a Ci-Cealkyl group, to yield the compound of formula (VIII):
which compound of formula (VIII) is subjected to a halogénation (using preferably Nbromosuccinimide or A-iodosuccinimide) to yield the compound of formula (IX):
(IX) wherein Hal represents a halogen atom, which compound of formula (IX) is further subjected to a Suzuki coupling reaction, in an aqueous or organic medium, in the presence of a palladium catalyst (preferably Pd(dppf)C12), of a base (preferably potassium carbonate), with a compound of formula (X):
to yield the compound of formula (XI):
the benzothiazole group of which compound of formula (XI) is deprotected (using preferably HFxPyr) and the ester function is hydrolysed (using preferably LiOHxH2O) to yield the compound of formula (I), which compound of formula (I) may be purified according to a conventional séparation 15 technique, which may be converted into its addition salts with a pharmaceutically acceptable acid or base and which is optionally separated into its isomers according to a conventional séparation technique, it being understood that, at any time considered appropriate in the course of the above-described process, hydroxy, amino, carboxylic and phosphono groups of the reagents or intermediates of synthesis may be protected and then deprotected according to the requirements of synthesis.
E37. Synthesis intermediate according to E34 or E36 which is:
wherein R4 and m are as defined in general formula (I), preferably R4 represents a hydrogen atom.
E38. The compound according to El wherein R4 represents a hydrogen, fluorine, chlorine or bromine atom, a methyl or a methoxy group.
E39. The compound according to El wherein Rs represents a group selected from: hydrogen; linear or branched Ci-Côalkyl, -NR’aR’b; -NR’a-CO-OR’c; -N+R’aR’bR’c; -O-R’c; -NH-X’2N+R’aR’bR’c; -O-XÙ-NR’aR’b, -NR’C-X’2-N3 and :
-NR'c—X'2-^CH
E41L The compound according to El wherein R’a and R’b independently of one another, represent a group selected from: hydrogen; heterocycloalkyl; -S02-phenyl wherein the phenyl may be substituted by a linear or branched Ci-Côalkyl; linear or branched Ci-Cealkyl optionally substituted by one or two hydroxyl groups; Ci-C6alkylene-SO2OH; Ci-Côalkylene-SChO'; Ci15 C6alkylene-COOH; Ci-C6alkylene-PO(OH)2; Ci-C6alkylene-NR’dR’e;
Ci-C6alkylene-N+R’dR’eR’f; Ci-Cealkylene-O-Ci-Cealkylene-OH; Ci-Côalkylene-phenyl wherein the phenyl may be substituted by a Ci-Cealkoxy group;
the group:
or R’a and R’b form with the nitrogen atom carrying them a cycle B3, or R’a, R’b and R’c form with the nitrogen atom carrying them a bridged Cs-Csheterocycloalkyl.
E41, Compound according to any of El to E31 wherein m=l.
Pharmacological study of the compounds of the invention has shown that they hâve proapoptotic properties. The ability to reactivate the apoptotic process in cells is of major therapeutic interest in the treatment of cancers and of immune and auto-immune diseases. In particular, the compounds according to the invention will be useful in the treatment of chemoor radio-resistant cancers.
In another embodiment, the compounds of the invention could be used for treating diseases or conditions characterized by an excess or a deregulated activity of platelets, especially prothrombotic conditions.
As used herein, the term “treat”, “treating or treatment of any disease or disorder refers in one embodiment, to ameliorating the disease or disorder (i.e., slowing or arresting or reducing 15 the development of the disease or at least one of the clinical symptoms thereof). In another embodiment “treat”, treating or treatment refers to alleviating or ameliorating at least one physical parameter including those which may not be discernible by the patient. In yet another embodiment, “treat”, treating or treatment refers to modulating the disease or disorder, either physically, (e.g., stabilization of a discernible symptom), physiologically, (e.g., 20 stabilization of a physical parameter), or both.
Among the cancer treatments envisaged there may be mentioned, without implying any limitation, the treatment of haematological malignancies and solid tumors. Haematological malignancies include myeloma, especially multiple myeloma, lymphoma, especially NonHodgkin Lymphoma (NHL) and more especially Diffuse Large B-cell Lymphoma (DLBCL), 25 and leukemia, especially Chronic Lymphocytic Leukemia (CLL), T-cell Acute Lymphoblastic
Leukemia (T-ALL), B-cell Acute Lymphoblastic Leukemia (B-ALL) and Acute Myelogenous Leukemia (AML). Solid tumors include the bladder, brain, breast, utérus, œsophagus and liver cancers, colorectal cancer, rénal 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 results from the leukemic transformation of thymie cell precursors and their arrest at spécifie stages of différentiation. Despite recent and extensive insights into the molecular and cellular mechanisms responsible for T-ALL onset and progression, this knowledge has not been translated into efficient targeted thérapies. Current clinical treatments include chemotherapy associated or not with hematopoietic stem cell transplantation with survival rates remaining around 50 and 70% in adult and pédiatrie cases, respectively. Both in pédiatrie and adult cases, relapses show very poor prognosis, reinforcing the need of the discovery of novel therapeutic options (Passaro et al., Immunol. Rev. 2016 May;271(l):15672). It has been shown that dual Bcl-2/Bcl-xL inhibitors, like ABT-263 and ABT-737, hâve promising activity in T-ALL patient derived xenograft models (Van Delft et al. Cancer Cell 2006;10:389-99; Suryani et al., Clin. Cancer Res. 2014,20:4520-31). Other studies hâve 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 sélective Bcl-xL inhibitor A-1331852 described previously hâve also shown to hâve in vitro and in vivo activity in the mature T-ALL cell line xenograft model MOLT-4 (Leverson et al., Sci. Transi. Med. 2015 Mar 18;7(279):279ra40). In a particular embodiment, tumor growth inhibition was also observed in MOLT-4 xenograft model upon treatment with the Bcl-xL inhibitors of the invention. These data support the use of the présent compounds in the treatment of T-ALL.
Among the treatments of autoimmune diseases envisaged there may be mentioned, without implying any limitation, the treatment of rheumatoid arthritis (RA) and systemic lupus erythematosus (SLE).
The présent invention relates also to pharmaceutical compositions comprising at least one compound of formula (I), as the active ingrédient, in combination with one or more pharmaceutically acceptable excipients. In particular, these pharmaceutical compositions are interesting for use as pro-apoptotic and/or anti-proliferative agents, particularly, in the treatment of cancers and of auto-immune and immune system diseases.
Suitable excipients according to the invention include diluents, lubricants, binders, disintegration agents, stabilisers, preservatives, absorbents, colorants, sweeteners and flavourings.
By way of non-limiting example there may be mentioned:
♦ as diluents: lactose, dextrose, sucrose, mannitol, sorbitol, cellulose, glycerol, * as lubricants: silica, talc, stearic acid and its magnésium and calcium salts, polyethylene glycol, ♦ as binders: magnésium aluminium silicate, starch, gelatin, tragacanth, methylcellulose, sodium carboxymethylcellulose and polyvinylpyrrolidone, ♦ as disintegrants: agar, alginic acid and its sodium sait, effervescent mixtures.
Among the pharmaceutical compositions according to the invention there may be mentioned more especially those that are suitable for oral, parentéral, nasal, per- or trans-cutaneous, rectal, perlingual, ocular or respiratory administration, especially tablets, dragées, sublingual tablets, capsules, glossettes, capsules, lozenges, injectable or drinkable préparations, aérosols, eye or 15 nose drops, suppositories, creams, ointments, dermal gels.
Actual dosage levels of the active ingrédients in the pharmaceutical compositions of this invention may be varied so as to obtain an amount of the active ingrédient which is effective to achieve the desired therapeutic response for a particular patient, composition, and mode of administration, without being toxic to the patient. The selected dosage level will dépend upon 20 a variety of factors including the activity of the particular compound of the présent invention employed, the route of administration, the time of administration, the rate of excrétion or metabolism of the particular compound being employed, the rate and extent of absorption, the duration of the treatment, other drugs, compounds and/or materials used in combination with the particular compound employed, the âge, sex, weight, condition, general health and prior 25 medical history of the patient being treated, and like factors well known in the medical arts.
A suitable daily dose of a compound of the invention will dépend upon the factors described above and may range from 0.01 mg to 2.5 g per day in one or more administration(s).
In another aspect, the présent invention relates also to the combination of a compound of formula (I) with an anticancer agent selected from genotoxic agents, mitotic poisons, anti métabolites, protéasome inhibitors, kinase inhibitors and antibodies, and also to pharmaceutical compositions comprising that type of combination and their use in the manufacture of médicaments for use in the treatment of cancer.
In another aspect, the compounds of the invention can be used in combination with radiotherapy 5 in the treatment of cancer.
Altematively, the compounds of the invention may be linked to monoclonal antibodies. Antibody Drug Conjugales (ADCs) represent a new class of therapeutics that is formed by chemically linking a cytotoxic drug to a monoclonal antibody through a linker. The monoclonal antibody of an ADC selectively binds to a target antigen of a cell (e.g. cancer cell) and releases 10 the drug into the cell. ADCs hâve therapeutic potential because they combine the specificity of the antibody and the cytotoxic potential of the drug. Nonetheless, developing ADCs as therapeutic agents has thus far met with limited success owing to a variety of factors such as unfavorable toxicity profiles, low efficacies and poor pharmacological parameters. Accordingly, there is still a need for new ADCs that overcome these problems and can 15 selectively deliver Bcl-xL to target cancer cells.
In another aspect, the compounds of the invention may be linked to fragments of monoclonal antibodies or linked to scaffold proteins that can be related or not to monoclonal antibodies. Antibody fragments must be understood as fragments of Fv, scFv, Fab, F(ab')2, F(ab'), scFv-Fc type or diabodies, which generally hâve the same specificity of binding as the antibody from 20 which they are descended. According to the présent invention, antibody fragments of the invention can be obtained starting from antibodies by methods such as digestion by enzymes, such as pepsin or papain, and/or by cleavage of the disulfide bridges by Chemical réduction. In another manner, the antibody fragments comprised in the présent invention can be obtained by techniques of genetic recombination likewise well known to the person skilled in the art or else 25 by peptide synthesis by means of, for example, automatic peptide synthesizers such as those supplied by the company Applied Biosystems, etc.
Scaffold proteins that can be related or not to monoclonal antibodies are understood to mean a protein that contains or not an immunoglobulin fold and that yields a binding capacity similar to a monoclonal antibody. The man skilled in the art knows how to select the protein scaffold. 30 More particularly, it is known that, to be selected, such a scaffold should display several features as follow (Skerra, J. Mol. Recogn. 2000, 13, 167-187): phylogenetically good conservation, robust architecture with a well-known three-dimensional molecular organization (such as, for example, crystallography or NMR), small size, no or only a low degree of post-translational modifications, easy to produce, express and purify. Such a protein scaffold can be, but without 5 limitation, a structure selected from the group consisting in fibronectin and preferentially the tenth fibronectin type III domain (FNfnlO), lipocalin, anticalin (Skerra, J. Biotechnol. 2001, 74, 257-75), the protein Z dérivative from the domain B of staphylococcal protein A, thioredoxin A or any protein with a repeated domain such as an “ankyrin repeat” (Kohl et al. PNAS 2003, 100, 1700-1705), “armadillo repeat”, “leucine-rich repeat” or “tetratricopeptide repeat”. There 10 could also be mentioned a scaffold dérivative from toxins (such as, for example, scorpion, insect, plant or mollusc toxins) or protein inhibitors of neuronal nitric oxide synthase (PIN).
The following Examples illustrate the invention but do not limit it in any way. All intermedia tes for preparing Examples are either commercially available or can be obtained by the person skilled in the art using conventional Chemical reactions described in the literature.
GENERAL PROCEDURES
Ail reagents obtained from commercial sources were used without further purification. Anhydrous solvents were obtained from commercial sources and used without further drying.
Column Chromatography
Automated flash column chromatography was performed on ISCO CombiFlash® Rf 200 or CombiFlash® Rf+ Lumen™ using RediSep® Rf Normal-phase Silica Flash Columns (35-70pm, 60 Â), RediSep Rf Gold® Normal-phase Silica High Performance Columns (20-40pm, 60 Â), RediSep® Rf Reversed-phase C18 Columns (40-63 pm, 60 Â), or RediSep Rf Gold® Reversedphase C18 High Performance Columns (20-40 pm, 100 Â).
TLC
Thin layer chromatography was conducted with 5 x 10 cm plates coated with Merck Type 60 F254 silica-gel.
Microwave Reactions
Microwave heating was performed with a CEM Discover® SP, or with an Anton Paar Monowave Microwave Reactor.
NMR ’H-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-dô or CDCI3 as solvent. 1H NMR data is in the form of delta values, given in part per million (ppm), using the residual peak of the solvent (2.50 ppm for DMSO-dô and 7.26 ppm for CDCI3) as internai standard. Splitting patterns are designated as: s (singlet), d (doublet), t (triplet), q (quartet), quint (quintet), sept (septet), m (multiplet), br s (broad 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 présent invention were characterized by high performance liquid chromatography-mass spectroscopy (HPLC-MS) on Agilent HP1200 with Agitent 6140 quadrupole LC/MS, operating in positive or négative ion electrospray ionisation mode. Molecular weight scan range is 100 to 1350. Parallel UV détection was done at 210 nm and 254 nm. Samples were supplied as a 1 mM solution in ACN, or in THF/H2O (1:1) with 5 pL loop injection. LCMS analyses were performed on two instruments, one of which was operated with basic, and the other with acidic eluents.
Basic LCMS: Gemini-NX, 3 pm, 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 from 100% Solvent A and finishing at 100% Solvent B over various/certain duration of time.
Acidic LCMS: KINATEX XB-C18-100A, 2.6pm, 50 mm*2.1 mm column at 40 °C, at a flow rate of 1 mL min-1 using 0.02% v/v aqueous formic acid (Solvent A) and 0.02% v/v formic acid in acetonitrile (Solvent B) with a gradient starting from 100% Solvent A and finishing at 100% Solvent B over various/certain duration of time.
Certain other compounds of the présent invention were characterized HPLC-MS under spécifie named methods as follows. For ail of these methods UV détection was by diode array detector at 230, 254, and 270 nm. Sample injection volume was 1 pL. Gradient elutions were run by defining flow rates and percentage mixtures of the following mobile phases, using ElPLC-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.
Rétention times (RT) for these named methods are reported in minutes. Ionisation is recorded in positive mode, négative mode, or positive-negative switching mode. Spécifie details for individual methods follow.
LCMS-V-B methods
Using an Agilent 1200 SL sériés instrument linked to an Agilent MSD 6140 single quadrupole with an ESI-APCI multimode source (Methods LCMS-V-B1 and LCMS-V-B2) or using an Agilent 1290 Infinity II sériés instrument connected to an Agilent TOF 6230 with an ESI-jet stream source (Method LCMS-V-B1); column: Thermo Accucore 2.6 pm, C18, 50 mm x 2.1 mm at 55 °C. Gradient details for methods LCMS-V-B1 and LCMS-V-B2:
Time (min) LCMS-V-B1 LCMS-V-B2 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
LCMS-V-C method
Using an Agilent 1200 SL sériés instrument linked to an Agilent MSD 6140 single quadrupole with an ESI-APCI multimode source; column: Agilent Zorbax Eclipse plus 3.5 pm, C18(2), 30 mm x 2.1 mm at 35 °C. Gradient details for method LCMS-V-C:
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
Préparative HPLC
Certain compounds of the présent invention were purified by high performance liquid chromatography (HPLC) on an Armen Spot Liquid Chromatography or Teledyne EZ system with a Gemini-NX® 10 μΜ C18, 250 mm x 50 mm i.d. column running at a flow rate of 118 mL min-1 with UV diode array détection (210 — 400 nm) using 25 mM aqueous NH4HCO3 solution and MeCN or 0.1% TFA in water and MeCN as eluents.
Certain other compounds of the présent invention were purified by HPLC under spécifie named methods as follows:
HPLC-V-A methods
These were performed on a Waters FractionLynx MS autopurification system, with a Gemini® 5 pm C18(2), 100 mm x 20 mm i.d. column from Phenomenex, running at a flow rate of 20 cm3min-1 with UV diode array détection (210ΜΌ0 nm) and mass-directed collection. The mass spectrometer was a Waters Micromass ZQ2000 spectrometer, operating in positive or négative ion electrospray ionisation modes, with a molecular weight scan range of 150 to 1000.
Method HPLC-V-A1 (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
Method HPLC-V-A2 (pH 9):
Solvent A: 10 mM aqueous ammonium acetate + 0.08% (v/v) conc. ammonia; Solvent B: acetonitrile + 5% (v/v) Solvent A + 0.08% (v/v) conc. ammonia
HPLC-V-B methods
Performed on an AccQPrep HP125 (Teledyne ISCO) System, with a Gemini® NX 5 pm C18(2), 150 mm x 21.2 mm i.d. column from Phenomenex, running at a flow rate of 20 cm-Wn1 with UV (214 and 254 nm) and ELS détection.
Method HPLC-V-B1 (pH 4):
Solvent A: water + 0.08% (v/v) formic acid; solvent B: acetonitrile + 0.08% (v/v) formic acid.
Method HPLC-V-B2 (pH 9):
Solvent A: water + 0.08% (v/v) conc. ammonia; solvent B: acetonitrile + 0.08% (v/v) conc. ammonia.
Method HPLC-V-B3 (neutral):
Solvent A: water; Solvent B; acetonitrile.
Analytical GC-MS
Combination gas chromatography and low resolution mass spectrometry (GC-MS) was performed on Agilent 6850 gas chromatograph and Agilent 5975C mass spectrometer using 15 m x 0.25 mm column with 0.25 pm HP-5MS coating and hélium as carrier gas. Ion source: EI+, 70 eV, 230°C, quadrupole: 150°C, interface: 300°C.
High-resolution MS
High-resolution mass spectra were acquired on an Agilent 6230 time-of-flight mass 5 spectrometer equipped with a Jet Stream electrospray ion source in positive ion mode.
Injections of 0.5μΐ were directed to the mass spectrometer at a flow rate 1.5 ml/min (5mM ammonium-formate in water and acetonitrile gradient program), using an Agilent 1290 Infinity HPLC system. Jet Stream parameters: drying gas (N2) flow and température: 8.0 1/min and 325 °C, respectively; nebulizer gas (N2) pressure: 30 psi; capillary voltage: 3000 V; sheath gas flow 10 and température: 325 °C and 10.0 1/min; TOFMS parameters: fragmentor voltage: 100 V;
skimmer potential: 60 V; OCT 1 RF Vpp:750 V. Full-scan mass spectra were acquired over the m/z range 105-1700 at an acquisition rate of 995.6 ms/spectrum and processed by Agilent MassHunter B.04.00 software.
Chemical naming lUPAC-preferred names were generated using ChemAxon’s ‘Structure to Name’ (s2n) functionality within MarvinSketch or J Chem for Excel (JChem versions 16.6.13 — 18.22.3), or with the Chemical naming functionality provided by Biovia® Draw 4.2.
1 Abbreviations
Ahx 6-hexanoic acid monomer
Boc tert-butyloxycarbonyl
BOC2O di-tert-butyl dicarbonate
5 ‘BuOH iert-butanol
cc. or conc. concentrated
CyOH cyclohexanol
dba ( 1E,4E)-1,5-diphenyIpenta-1,4-dien-3-one, dibenzylideneacetone
DCM dichloromethane
10 DEA diethanolamine
DIAD diisopropylazodicarboxylate
DIBAL-H diisobutylaluminium hydride
DIPA A-isopropylpropan-2-amine, diisopropylamine
DIPEA A-ethyl-A-isopropyl-propan-2-amine, diisopropylethylamine
15 EtO2 diethyl ether
EtOAc ethyl acetate
eq. équivalent
HFxPyr Hydrogen fluoride pyridine
hs homo sapiens
20 LDA lithium diisopropylamide
MeCN acetonitrile
MeOH methanol
MTBE methyl tert-butyl ether
on overnight
25 Pd\C palladium on carbon
PPh3 triphenylphosphine
• rt room température
RT rétention time (in minutes)
TB AF tetrabutylammonium fluoride
TBAOH tetrabutylammonium hydroxide
5 TBSC1 tert-butyl-chloro-dimethyl-silane
TEA ••-diethylethanamine
TFA 2,2,2-trifluoroacetic acid
pTSA 4-methylbenzenesulfonic acid
THF tetrahydrofuran
10 TIPSC1 chloro(triisopropyl)silane
Xantphos 4,5-Bis(diphenylphosphino)-9,9-dimethylxanthene
Xantphos Pd G3 [(4,5-Bis(diphenylphosphino)-9,9-dimethylxanthene)-2-(2'-amino-l,rbiphenyl)]palladium(II) methanesulfonate
BINAP 2,2 '-Bis(dipheny Iphosphino)-1,1 '-binaphthy 1
15 rac-BINAP Pd G3 [(2,2'-Bis(diphenylphosphino)-1,1 '-binaphthy l)-2-(2'-amino-1,1'biphenyl)]palladium(II) methanesulfonate
Pd(dppf)Cl2.CH2C12 [1,1 '-Bis(diphenylphosphino)ferrocene]dichloropalladium(II)
Pd2(dba)3 Tris(dibenzylideneacetone)dipalladium(0)
Pd(PPh3)2C12 Bis(triphenylphosphine)palladium chloride
20 Pd(AtaPhos)2C12 bis(di-tert-butyl(4-
dimethylaminophenyl)phosphine)dichloropalladium(II)
Named General Procedures
The following are représentative experimental procedures that are referred to by name in subséquent Préparations.
Somogashira General Procedure
The mixture of 1 eq. of aryl halogenide, 2 eq. of acetylene, 0.05 eq. of Pd(PPh3)2Cl2, 0.05 eq. of Cul, and DIPA (1 mL/mmol) in THF (5 mL/mmol) was kept at 60 °C. After reaching an appropriate conversion the volatiles were removed under reduced pressure, the crude intermediate was purified via flash column chromatography.
Deprotection with HFIP General Procedure
Substrate in l,l,l,3,3,3-hexafluoropropan-2-ol (10 mL/mmol) was kept at 100-120 °C in a pressure bottle. After reaching an appropriate conversion the volatiles were removed under reduced pressure, the crude intermediate was purified via flash column chromatography.
Alkylation General Procedure
The mixture of 1 eq. of phenol/carbamate, 1-2 eq. of alkyl iodide or bromide, and 2-3 eq. of 15 Cs2CO3 in acetone (5 mL/mmol) was stirred at rt for phénols and at 55 °C for carbamates. After reaching an appropriate conversion the volatiles were removed under reduced pressure, the crude intermediate was purified via flash column chromatography or reverse phase flash column chromatography.
Mitsunobu General Procedure
To the mixture of 1.0-1.5 eq. of aliphatic alcohol, 1 eq. of carbamate/phenol, and 1-2 eq. triphenylphosphine in THF or toluene (5 mL/mmol) was added 1-3 eq. of ditertbutyl azodicarboxylate / diisopropyl azodicarboxylate in one portion. The mixture was stirred at rt or 50 °C, if necessary, for the carbamate and at rt for the phénol. After reaching an appropriate conversion the volatiles were removed under reduced pressure, the crude intermediate was 25 purified via flash column chromatography.
Buchwald General Procedure I
The mixture of 1 eq. of chloro-substrate, 2 eq. of l,3-benzothiazol-2-amine, 0.1 eq. of Pd2(dba)3, 0.2 eq. of XantPhos and 3 eq. of DIPEA in CyOH (5 mL/mmol) was kept at 140 °C. After reaching an appropriate conversion, the reaction mixture was diluted with DCM (10 mL/mmol), injected onto a preconditioned silica gel column and was purified via flash column chromatography.
Buchwald General Procedure II
The mixture of 1 eq. of thiazol amine, 1.2-1.5 eq. of (Z)-N-(6-chloro-4-methyl-pyridazin-3-yl)3-(2-trimethylsilylethoxymethyl)-l,3-benzothiazol-2-imine, 3 eq. of CS2CO3, 0.1 eq. of Pd2(dba)3, 0.2 eq. of XantPhos and 3 eq. of DIPEA in 1,4-dioxane (5 mL/mmol) was kept at reflux. After reaching an appropriate conversion the volatiles were removed under reduced pressure, the crude intermediate was purified via flash column chromatography.
Finkelstein General Procedure
The mixture of 1 eq. of alkyl chloride and 2 eq. of Nal in acetone (5 mL/mmol) was kept at reflux. After reaching an appropriate conversion the volatiles were removed under reduced pressure, the crude intermediate was purified via flash column chromatography.
Nucleophile Substitution General Procedure
The mixture of 1 eq of thiazol amine, 1 eq. of 3,6-dichloro-4-methylpyridazine, and 1.5 eq. of CS2CO3 in toluene (4 mL/mmol) was kept at 150 °C. After reaching an appropriate conversion the volatiles were removed under reduced pressure, the crude intermediate was purified via flash column chromatography.
Alkylation with tosylate General Procedure
An oven-dried vial was equipped with a PEEE-coated magnetic stirring bar, and was charged with 1 eq. tosylate and 5 eq. as the appropriate amine were suspended in MeCN (5 mL/mmol). The reaction mixture was then warmed up to 50 °C and stirred at that température until no further conversion was observed. The reaction mixture was diluted with DCM then it was injected onto a DCM preconditioned silica gel column. Then it was purified via flash column chromatography using DCM and MeOH (1.2% NH3) as eluents.
Alkylation with in situ generated tosylate General Procedure
An oven-dried vial was equipped with a PEEE-coated magnetic stirring bar, and was charged with 1 eq. of hydroxy dérivative, 3 eq. of JV,A-diethylethanamine, catalytic amount of N,Ndimethylpyridin-4-amine and DCM (5 mL/mmol) then 2 eq. of 4-methylbenzenesulfonyl chloride was added in one portion and stirred at rt until no further conversion was observed (typically >90%, 1-5 h). After the evaporatation of the volatiles with N2 at 40 °C, 30 eq. of the appropriate primary, secondary, or tertiary amine in acetonitrile (3 mL/mmol) were added and the mixture was stirred at 50 °C until full conversion was observed regarding the tosyl denvative (typically 1-3 h). Reaction mixture was concentrated onto celite and purified via flash column chromatography using EtOAc and MeOH (1.2% NH3) as eluents or reverse phase flash column chromatography (0.1% TFA in water : MeCN).
Alkylation with in situ generated iodine General Procedure
An oven-dried vial was equipped with aPTFE-coated magnetic stirring bar, it was charged with 2 eq. PPh3 and 2 eq. imidazole then DCM (5 mL/mmol) was added. To the resulting mixture 2 eq. iodine was added portionwise then stirred for 15 min at rat. To the resulting mixture 1 eq. of the appropriate alcohol was added dissolved in DCM and stirred at rt until no further conversion was observed. To the generated iodo compound 5 eq. of the appropriate amine was added and then stirred for 30 min at rt, while full conversion was observed. Celite was added to the reaction mixture and the volatiles were removed under reduced pressure. Then it was purified via flash chromatography using DCM and MeOH (1.2% NH3) eluents.
Alkylation of Silyl-Protected Phénols General Procedure
The mixture of 1 eq. of silyl-protected phénol, 1 eq. of alkyl iodide, and 1.15 eq. of TB AF (1 M in THF) in THF (2 mL/mmol) was stirred at rt. After reaching an appropriate conversion the volatiles were removed under reduced pressure, the crude intermediate was purified via flash column chromatography.
Deprotection and Hydrolysis General Procedure
The mixture of 1 eq. of substrate and 100 eq. of HFxPyr in MeCN (15 mL/mmol) was stirred at 60 °C. After reaching an appropriate conversion, the 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. of LiOHxHîO, and stirred at 60 °C. After reaching an appropriate conversion, the volatiles were removed under reduced pressure and the crude product was purified via flash column chromatography using DCM and MeOH (containing 1.2% NH3) as eluents.
Quaternary sait formation General Procedure
An oven-dried vial was equipped with a PTFE-coated magnetic stirring bar, and was charged with 1 eq. tosylate and 20 eq. as the appropriate amine were suspended in CyOH (5 mL/mmol). The reaction mixture was then warmed up to 140°C and stirred at that température until no further conversion was observed. The reaction mixture was diluted with DCM then it was injected onto a DCM preconditioned silica gel column. Then it was purified via flash column chromatography using DCM and MeOH (1.2% NH3) as eluents.
Quaternary sait deprotection General Procedure
To a THF (5 mL/mmol) solution of the appropriate quaternary sait 3 eq. TBAF was added, and then it was stirred at rt until no further conversion was observed. The reaction mixture was the evaporated to dry under reduced pressure. To a suspension of 1 eq. desilylated quaternary sait in dry MeCN (15 mL/mmol), 100 eq. of HFxPyr added, and then was stirred at 60°C. After reaching an appropriate conversion, the volatiles were removed under reduced pressure, the residue was suspended in a 1:1 mixture of THF - water (30 mL/mmol), 150 eq. of LiOH x H2O was added, and the mixture was stirred at rt. After reaching an appropriate conversion, the volatiles were removed under reduced pressure. The crude product was purified via flash column chromatography using DCM and MeOH (containing 1.2% NH3) as eluents.
Alkylation, Deprotection and Hydrolysis General procedure
A mixture of tertiary amine (1 eq.) and alkylating agent (10 eq.) in acetonitrile (3 mL/mmol) was stirred at rt. After reaching appropriate conversion, the volatiles were removed under reduced pressure and purified via reverse phase flash column chromatography, if it was necessary, otherwise 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 appropriate 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), LiOHxHzO (150 eq.) was added and the mixture was stirred at 60°C. After reaching appropriate conversion to the desired product, the volatiles were removed under reduced pressure and the crude product was purified via reverse phase flash column chromatography.
Deprotection of terf-butyl-dimethyl-silyl protecting group General Procedure
The appropriate TBS protected compound and [(15,4jR)-7,7-dimethyl-2-oxo-norbornan-lyl]methanesulfonic (0.25 eq. in general, 1.25 eq. if A,/V-dimethyl-amine was presented in the molécule) acid were mixed in the mixture of methanol (2.5 mL/mmol) and dichloromethane (2.5 mL/mmol) in a sealed tube and stirred at 50°C until full conversion was observed (typically 1-5 h). After the reaction time the mixture was poured into a séparation funnel, diluted with DCM and washed with cc. NaHCO3 and water. Dried over MgSO4, filtered and onto celite then purified via flash column chromatography.
Deprotection and hydrolysis of phosphonic acid dérivatives General Procedure
The solution of the diethyl phosphonate dérivative in DCM (5 mL/mmol) and MeCN (5mL/mmol) was flushed with nitrogen then 40 eq. TEA was added and then cooled to 0°C. 20 eq. bromo(trimethyl)silane was added in one portion then stirred for further 30 min at 0°C. Left to warm up to rt and stirred until the full conversion regarding the ethyl ester (typically 1-5 h). Boc protecting group(s) were generally but only partially eliminated, resulted in the mixture of products. After full conversion, the mixture was quenched with few mL MeOH at 0°C, concentrated to Celite then purified via flash column chromatography using EtOAc and MeOH (containing 1.2% NH3) as eluents. The two products (with Boc, without Boc) were combined and continued with Deprotection and Hydrolysis General Procedure.
Amine substitution and Hydrolysis General procedure
To the product from Préparation 14 in a 1:1 mixture of acetonitrile and N-methyl-2pyrrolidone (10 ml/mmol), was added the appropriate amine (3-10 eq), and the mixture was stirred at 50 °C for 2-24 h. After the addition of 70% HF in pyridine (50-100 eq) at rt, the mixture was stirred for 4-18 h. After the 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 LiOHxHzO (5 eq) was added, and stirred at 2040 °C for 1-4 h. The hydrolysed product was purified by préparative HPLC (using acetonitrile and 5 mM aqueous NH4HCO3 solution as eluents) to give the desired product.
Préparations
The following experimental details describe the préparation of synthetic intermediates.
Préparation la: Methyl 2-{[(for/-butoxy)carbonyl]amino}-5-[3-(2-fluoro-4iodophenoxy)propyl]-l,3-thiazole-4-carboxylate
Step A: methyl2-(tert-butoxycarbonylamino)-5-iodo-thiazole-4-carboxylate
50.00 g of methyl 2-(tert-butoxycarbonylamino)thiazole-4-carboxylate (193.55 mmol, 1 eq.) was suspended in 600 mL dry MeCN. 52.25 g oîN-iodo succinimide (232.30 mmol, 1.2 eq.) was added and the resulting mixture was stirred overnight at room température. The reaction mixture was diluted with saturated brine, then it was extracted with EtOAc. The combined organic layers were extracted with 1 M Na2S2Os, then with brine again. Then dried over Na2SO4, filtered and the filtrate was concentrated under reduced pressure. The crude product was purified via flash column chromatography using heptane as eluent to obtain 60 g (156 mmol, 80%) of the desired product.
Ή NMR (400 MHz, DMSO-d6) δ ppm 12.03/11.06 (br s), 3.78 (s, 3H), 1.47 (s, 9H); 13C NMR (400 MHz, DMSO-d6) δ ppm 153.8, 82.5, 77.7, 52.3, 28.3; HRMS-ESI (m/z): [M+H]+ calcd for CioHi4lN204S: 384.9713, found 384.9708.
Step B: methyl 2-(tert-butoxycarbonylamino)-5-(3-hydroxyprop-l-ynyl)thiazole-4carboxylate
A 500 mL oven-dried, one-necked, round-bottom flask was equipped with a PTFE-coated magnetic stirring bar and fitted with a reflux condenser. It was charged with 9.6 g of the product from Step A (25 mmol, 1 eq.), 2.80 g of prop-2-yn-l-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 flushed with argon. After 5 minutes stirring under inert atmosphère 549 mg of Pd(PPh3)2Cl2 (1.25 mmol, 0.05 eq.) and 238 mg of Cul (1.25 mmol, 0.05 eq.) was added. The resulting mixture was then warmed up to 60°C and stirred at that température until no further conversion was observed. Celite was added to the reaction mixture and the volatiles were removed under reduced pressure. Then it was purified via flash column chromatography using heptane and EtOAc as eluents to give 7.30 g (23 mmol, 93%) of the desired product as a yellow solid.
XH NMR (400 MHz, DMSO-d6) δ ppm 12.10 (br s, 1H), 5.45 (t, 1H), 4.36 (d, 2H), 3.79 (s, 3H), 1.48 (s, 9H); 13C NMR (125 MHz, DMSO-d6) δ ppm 161.3, 142.4, 118.1, 101.4, 73.9, 52.4,
50.2, 28.3; HRMS-ESI (m/z): [M+H]+ calcd for C13H17N2O5S: 313.0853, found 313.0866.
Step C: methyl 2-(tert-butoxycarbonylamino)-5-(3-hydroxypropyl)thiazole-4-carboxylate
An 1 L oven-dried pressure bottle equipped with a PTFE-coated magnetic stirring bar was charged with 44.75 g of the product from Step B (143.3 mmol, 1 eq.), 7.62 g of Pd/C (7.17 5 mmol, 0.05 eq.) in 340 mL of éthanol, and then placed under a nitrogen atmosphère using hydrogénation System. After that it was filled with 4 bar H2 gas and stirred at rt overnight. Full conversion was observed, but only the olefin product was formed. After filtration of the catalyst through a pad of Celite the whole procedure was repeated with 5 mol% new catalyst. The resulting mixture was stirred overnight to get full conversion. Celite was added to the reaction 10 mixtures and the volatiles were removed under reduced pressure. Then it was purified via flash column chromatography using heptane and EtOAc as eluents to give 31.9 g (101 mmol, 70%) of the desired product as light-yellow crystals
Ή NMR (500 MHz, DMSO-d6) 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); 13C NMR (125 MHz, DMSO-d6) δ ppm 162.8, 15 143.1, 135.4, 60.3, 51.9, 34.5, 28.3, 23.4; HRMS-ESI (m/z): [M+H]+ calcd for C13H21N2O5S:
317.1166, found 317.1164.
Step D: methyl 2-{[(tert-butoxy)carbonyl]amino}-5-[3-(2-fluoro-4-iodophenoxy)propyl]-l,3thiazole-4-carboxylate
A 250 mL oven-dried, one-necked, round-bottomed flask equipped with a PTFE-coat.ed 20 magnetic stirring bar, was charged with 3.40 g of 2-fluoro-4-iodo-phenol (14 mmol, 1 eq.), 5.00 g of the product from Step C (16 mmol, 1.1 equiv) and 4.10 g of PPh3 (16 mmol, 1.1 eq.) and 71 mL of dry toluene. After 5 min stirring under nitrogen atmosphère, 3.10 mL of DI AD (3.20 g, 16 mmol, 1.1 eq.) was added in one portion while the reaction mixture warmed up. Then the reaction mixture was heated up to 50°C and stirred at that température for 30 min, when the 25 reaction reached complété conversion. The reaction mixture was directly injected onto a preconditioned silica gel column, and then it was purified via 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 desired product.
Ή NMR (500 MHz, DMSO-d6) δ ppm 11.64 (br s, 1H), 7.59 (dd, 1H), 7.45 (dd, 1H), 6.98 (t, 30 1H), 4.06 (t, 2H), 3.73 (s, 3H), 3.22 (t, 2H), 2.06 (m, 2H), 1.46 (s, 9H); 13C NMR (125 MHz,
DMSO-d6) δ ppm 134.0, 124.9, 117.6, 68.2, 51.9, 30.5, 28.3, 23.2; HRMS-ESI (m/z): [M+H]+ calcd for C19H23N2O5FSI: 537.0351, found 537.0348.
Préparation 1b: Methyl 2-(tert-butoxycarbonylamino)-5-[3-[4- [3-[tertbutoxycarbonyl(methyl)amino]prop-l-ynyl]-2-fluoro-phenoxy]propyl]thiazole-4carboxylate
A 500 mL oven-dried, one-necked, round-bottom flask was equipped with a PTFE-coated magnetic stirring bar and fitted with a reflux condenser. It was charged with 13.41 g of Préparation la (25 mmol, 1 eq.), 8.46 g of tert-butyl N-methyl-N-prop-2-ynyl-carbamate (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 flushed with argon. After 5 minutes stirring under inert atmosphère 549 mg of Pd(PPh3)2Ch (1.25 mmol, 0.05 eq.) and 238 mg of Cul (1.25 mmol, 0.05 eq.) were added. The resulting mixture was then warmed up to 60°C and stirred at that température until no further conversion was observed. Celite was added to the reaction mixture and the volatiles were removed under reduced pressure. Then it was purified via flash column chromatography using heptane and EtOAc as eluents to give 10.5 g (18.2 mmol, 73%) of the desired product.
Ή NMR (500 MHz, DMSO-d6) 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); 13C NMR (125 MHz, DMSO-d6) δ ppm 129.1, 119.2, 115.4, 68.1, 51.9, 38.6, 33.8, 30.5, 23.2; HRMS-ESI (m/z): [M+H]+ calcd for C28H37FN3O7S: 578.2331, found 578.2331.
Préparation le: Methyl 2-(tert-butoxycarbonylamino)-5-[3-[4-[3(dimethylamino)prop-l-ynyl]-2-fluoro-phenoxy]propyl]thiazole-4-carboxylate
A 250 mL oven-dried, one-necked, round-bottom flask was equipped with a PTFE-coated magnetic stirring bar and fitted with a reflux condenser. It was charged with 5.36 g of Préparation la (10 mmol, 1 eq.), 1.66 g oîN,N-dimethylprop-2-yn-l-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 flushed with argon. After 5 minutes stirring under inert atmosphère 220 mg of Pd(PPh3)2Ch (0.5 mmol, 0.05 eq.) and 95 mg of Cul (0.5 mmol, 0.05 eq.) were added. The resulting mixture was then warmed up to 60°C and stirred at that température until no further conversion was observed. Celite was added to the reaction mixture and the volatiles were removed under reduced pressure. Then it was purified via flash column chromatography using DCM and MeOH (1.2% NH3) as eluents to give 4.5 g (7.8 mmol, 78%) of the desired product.
Ή NMR (500 MHz, DMSO-d6) 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);
13C NMR (125 MHz, DMSO-d6) 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-ESI (m/z): [M+H]+ calcd for C24H31FN3O5S: 492.1963, found 492.1956.
Préparation 2a: 5-[terLButyl(dimethyl)silyl]oxy-4-[terLbutyl(diphenyl)silyl]oxypentan-l-ol
Step A; pent-4-enyl benzoate
30.00 g of pent-4-en-l-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 via dropping funnel under inert atmosphère. After the addition the mixture was further stirred at 0°C for 30 min then at rt for on. The mixture was diluted with 100 mL of DCM then the organic phase was washed with water, 1 M NaOH, 1 M HCl, brine, respectively. The organic phase was dried over MgSO4, filtered, concentrated and purified via flash column chromatography using heptane and EtOAc as eluents to give 63.19 g (95%) of the desired product as colorless liquid.
’H NMR (500 MHz, DMSO-d6) δ ppm 7.97 (dd, 2H), 7.66 (t, 1H), 7.53 (t, 2H), 5.91-5.81 (m, 1H), 5.09-4.97 (m, 2H), 4.27 (t, 2H), 2.17 (q, 2H), 1.81 (qv, 2H); 13C NMR (125 MHz, DMSOd6) 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]+ calcd for C12H14O2: 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.), 50.40 g of 4-methyl-4-oxido-morpholin4-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 tetraoxoosmium (2.5 w% in 2-methylpropan-2-ol, 0.64 mmol, 0.002 eq.) was added and the mixture was stirred at 60°C for 24 h. Full conversion was observed. The mixture was cooled down to rt and 1 M Na2S2O3 was added then stirred for further 10 min at rt. DCM was added and the organic phase was separated, washed with water, brine, respectively. The solution was dried over over MgSO4, filtered, concentrated and purified via flash column chromatography using heptane and EtOAc as eluents to give 36.9 g (63%) of the desired product as white solid.
Ή NMR (500 MHz, DMSO-d6) δ ppm 7.99-7.50 (m, 5H), 4.50 (m, 2H), 4.28 (m, 2H), 3.45 (m, 1H), 3.30-3.24 (m+m, 2H), 1.85-1.72 (m+m, 2H), 1.59-1.33 (m+m, 2H); 13C NMR (125 MHz, DMSO-dô) δ ppm 166.2, 133.8-129.1, 71.2, 66.3, 65.5, 30.3, 25.2; HRMS-ESI (m/z): [M+Na]+ calcd for Ci2Hi6NaO4: 247.0941, found 247.0941.
M Step C: 5-[tert-butyl(dimethyl)silyl]oxy-4-hydroxy-pentyl] 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 N,N-dimethylformamide then cooled to -20°C under inert atmosphère.
16.71 g of tert-butyl-chloro-dimethyl-silane (0.11 mol, 1 eq.) in 40 mL of N,N5 dimethylformamide was added in slow rate over a period of 30 min, supported with 10 mL of
DCM then left to warm up to rt and further stirred for on. Full conversion was observed. Quenched with cc. NH4CI then evaporated most of the volatiles. EtOAc and water were added to the residue, the organic phase was separated then washed with water and brine, dried over MgSÜ4, filtered, concentrated and purified via flash column chromatography using heptane and 10 EtOAc as eluents to give 33.71 g (90%) of the desired product as colorless oil.
Ή NMR (500 MHz, DMSO-d6) 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); 13C NMR (125 MHz, DMSO-46) 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.
StepD: [5-[tert-butyl(dimethyl)silyl]oxy-4-[tert-butyl(diphenyl)silyl]oxy-pentyl] 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,N-dimethylpyridin-4-amine (0.01, 0.1 eq.) were mixed in 230 mL of N,Ndimethylformamide then 38 mL of tert-butyl-chloro-diphenyl-silane (0.15 mol, 1.5 eq.) was added in slow rate, supported with 20 mL of N,N-dimethylformamide then stirred at 50°C for 20 overnight. Full conversion was observed. The mixture was cooled to rt, quenched with cc.
NH4CI then evaporated most of the volatiles. EtOAc and water were added to the residue, the organic phase was separated then washed with water and brine, dried over MgSÛ4, filtered, concentrated and purified via flash column chromatography using heptane and EtOAc as eluents to give 56.43 g (99%) of the desired product as colorless thick oil.
'H NMR (500 MHz, DMSO-d6) δ ppm 7.91-7.37 (m, 15H), 4.17 (m, 2 H), 3.76 (m, 1 H), 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); 13C NMR (125 MHz, DMSO-î/6) 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-ESI (m/z): [M+Na]+ calcd for C34H48NaO4Si2: 599.2983, found 599.2981.
Step E: 5-[tert-butyl(dimethyl)silyl]oxy-4-[tert-butyl(diphenyl)silyl]oxy-pentan-l-ol
46.10 g of the product from Step D (0.08 mol, 1 eq) was dissolved in 227 mL of MeOH and
117 mL of THF then 12.79 g of NaOH (0.32 mol, 4.0 eq.) in 85 mL of water was added slowly while the mixture was cooled with ice. After the addition the mixture left to stir at rt until full conversion was observed (ca. 4 h). EtOAc and water were added then separated and the organic phase was washed with brine, dried over MgSO4, filtered, concentrated and purified via flash column chromatography using heptane and EtOAc as eluents to give 29.32 g (78%) of the desired product as colorless oil.
Ή NMR (500 MHz, DMSO-Jg) 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); 13C NMR (125 MHz, DMSO-d6) 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; HRMS-ESI (m/z): [M+Na]+ calcd for C27H44NaO3Si2: 495.2721, found 495.2706.
Préparation 2b: 5-[ierAButyl(dimethyl)silyl]oxy-4-methoxy-pentan-l-ol
Step A: [5-[tert-butyl(dimethyl)silyl]oxy-4-methoxy-pentyl] benzoate
3.66 g of Préparation 2a, Step C (10.8 mmol, 1 eq), 6.95 g of N1,N1,N8,N8-
O tetramethylnaphthalene-l,8-diamine (32.4 mmol, 3 eq.) and a small portion 4A molecular sieves were mixed in 210 mL of DCM then 4.00 g of trimethyloxomum;tetrafluoroborate (27.0 mmol, 2.5 eq.) was added in one portion and the mixture was stirred at rt for 5 h. Full conversion was observed. The mixture was filtered through a pad of Celite, washed with DCM then concentrated. The leftover was redissolved in DCM and washed with water, 1 M CuSO4, brine, respectively. The solution was dried over MgSO4, filtered, concentrated onto Celite and purified via flash column chromatography using heptane and EtOAc as eluents to give 2.58 g (68%) of the desired product as colorless oil.
'H NMR (400 MHz, DMSO-d6) 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); 13C NMR (100 MHz, DMSO-d6) 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-methoxy-pentan-l-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 added slowly to the mixture. After the addition the mixture was stirred at rt until full conversion was observed (ca. 1.5 h). EtOAc and water were added then separated and the organic phase was washed with brine, dried over MgSO4, filtered and concentrated to give 1.76 g (98%) of the desired product as light yellow ’H NMR (400 MHz, DMSO-d6) 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); 13C 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): 5 [M+H]+ calcd for C12H29O3S1: 249.2, found 249.2.
Préparation 2c: 5-Methoxy-4-triisopropylsilyloxy-pentan-l-ol
Step A: [5-[tert-butyl(dimethyl)silyl]oxy-4-triisopropylsilyloxy-pentyl] benzoate
2.0 g of Préparation 2a, Step C (5.91 mmol, 1 eq) and 1.21 g of imidazole (17.73 mmol, 3 eq.) were mixed in 6 mL οΐΝ,Ν-dimethylformamide then 1.71 g of chloro(triisopropyl)silane (8.86 mol, 1.5 eq.) was added in one portion then stirred at 60°C for 3.5 h. Additional chloro(triisopropyl)silane (0.5 eq.) and imidazole (1 eq.) were added and stirred further 3 h. Full conversion was observed. The mixture was cooled to rt then EtOAc and water were added, the organic phase was separated, washed with brine, dried over MgSCL, filtered, concentrated onto Celite and purified via flash column chromatography using heptane and EtOAc as eluents 15 to give 2.27 g (99%) of the desired product as colorless thick oil.
Ή NMR (400 MHz, DMSO-d0) 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-triisopropylsilyloxy-pentyl) benzoate
Using Deprotection of tert-butyl-dimethyl-silyl protecting group General Procedure starting from 2.25 g of the product from Step A (4.54 mmol, 1 eq.) followed by purification via flash column chromatography using heptane and EtOAc as eluents, 1.13 g (65%) of the desired product was obtained.
'H NMR (400 MHz, DMSO-rfe) 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-MS25 ESI (m/z): [M+H]+ calcd for C21H37O4S1: 381.2, found 381.3.
Step C: (5-methoxy-4-triisopropylsilyloxy-pentyl) benzoate
1.11 g of the product from Step B (2.91 mmol, 1 eq) and 1.87 g of N1,N1,N8,N8tetramethylnaphthalene-l,8-diamine (8.7 mmol, 3 eq.) were mixed in 45 mL of DCM then 1.87 g of triniethyloxonium;tetrafluoroborate (8.7 mmol, 3 eq.) was added in one portion and the 30 mixture was stirred at rt for 4 h. Full conversion was observed. The mixture was filtered through a pad of Celite, washed with DCM then the organic phase was washed with water, 1 M CuSO4, brine, respectively. The solution was dried over MgSCU, filtered, concentrated onto Celite and purified via flash column chromatography using heptane and EtOAc as eluents to give 963 mg (84%) of the desired product as colorless oil.
Tl NMR (400 MHz, DMSO-rfe) δ ppm 7.95 (d, 2H), 7.66 (t, 1H), 7.52 (t, 2H), 4.28 (t, 2H), 5 3.96 (qv, 1H), 3.31 (d, 2H), 3.25 (s, 3H), 1.82-1.54 (m, 4H), 1.01 (m, 21H); 13C NMR (100
MHz, DMSO-de) 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-triisopropylsilyloxy-pentan-l-ol
953 mg of the product from Step C (2.41 mmol, 1 eq) was dissolved in 12 mL of MeOH and 6 10 mL of THF then 487 mg of NaOH (12.1 mmol, 5.0 eq.) in 6 mL of water was added slowly.
After the addition the mixture left to stir at rt until full conversion was observed (ca. 2 h). EtOAc and water was added then separated and the organic phase was washed with brine, dried over MgSÛ4, filtered, concentrated onto Celite and purified via flash column chromatography using heptane and EtOAc as eluents to give 687 mg (78%) of the desired product as colorless oil.
’H NMR (500 MHz, DMSO-d6) 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); 13C NMR (125 MHz, DMSO-d6) δ ppm 76.8, 71.4, 61.4, 58.8, 31.4, 28.3, 18.5, 12.5; HRMS-ESI (m/z): [M+H]+ calcd for Ci5H35O3Si: 291.2350, found 291.2349.
Préparation 2d: 4-[tert-Butyl(dimethyl)siIyl]oxy-5-(dimethylamino)pentan-l-ol
Step A: 3-(oxiran-2-yl)propyl benzoate
19.01g of 3-chlorobenzenecarboperoxoic acid (77w%, 85.83 mmol, 1.3 eq.) was dissolved in DCM (3 mL/mmol) in a séparation funnel then the organic phase (3-4 mL water was excluded) was added directly to the solution of 12.41 g of Préparation 2a, Step A (65.25 mmol, 1 eq.) in DCM (3 mL/mmol) and stirred at rt for 4 h. Full conversion was observed. 100 mL of DCM 25 was added then washed with 2x100 mL cc. NaHCO3 and 1x100 mL brine, dried over MgSO4, filtered, concentrated and purified via flash column chromatography using heptane and EtOAc as eluents to give 13.92 g (quant.) of the desired product as colorless oil.
’H NMR (400 MHz, DMSO-d6) 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); 13C NMR (100 30 MHz, DMSO-ί/ό) δ ppm 166.2, 133.8, 129.6, 129.3, 64.8, 51.7, 46.6, 29.1, 25.4; HRMS-EI (m/z): [M]+ calcd for Ci2Hi4O3: 206.0943, found 206.0941.
Step B: [4-[tert-butyl(dimethyl) silyljoxy-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 mml, 3 eq.) were mixed in a sealed tube and stirred at 60°C for 1 h. Full conversion was observed. The reaction mixture was concentrated and [5-(dimethylamino)-4hydroxy-pentyl] benzoate was obtained as a thick light yellow oil. HRMS-ESI (m/z): [M+H]+ calcd for C14H22NO3: 252.1594, found 252.1597.
The crude product was redissolved in N,N-dimethylformamide (2 mL/mmol) then 6.95 g of imidazole (102 mmol, 3 eq.), 208 mg of N,N-dimethylpyridin-4-amine (1.70 mmol, 0.05 eq.) were added and after getting clear solution 12.8 g of tert-butyl-chloro-dimethyl-silane (85.0 mmol, 2.5 eq.) was added in one portion. The mixture was stirred at 60°C for 3 h (>95% conversion was observed). Cooled down to rt then 3-4 mL cc. NH4CI was added, stirred for 5 mins then the volatiles were evaporated. 300 mL of EtOAc, 50 mL of water and 50 mL cc. NaHCO3 were added then the organic layer was separated. Organic phase was washed with 1x50 mL H2O, 1x50 mL brine then dried over MgSO4, filtered then concentrated. The crude product was purified via flash column chromatography using DCM and MeOH as eluents to give 8.37 g (67%) of the desired product as colorless oil.
Ή NMR (400 MHz, DMSO-d6) 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); 13C NMR (100 MHz, DMSO-î/6) δ ppm 166.2, 130.3, 70.0, 66.0, 65.3, 46.6, 32.0, 26.3, 24.5, -3.8, -4.4; HRMS-ESI (m/z): [M+H]+ calcd for C2oH36N03Si: 366.2459, found 366.2463.
Step C: 4-[tert-butyl(dimethyl) silyljoxy-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 added slowly. After the addition the mixture left to stir at rt until full conversion was observed (ca. 1.5 h). EtOAc and water was added then separated and the organic phase was washed with brine, dried over MgSÛ4, filtered, concentrated onto Celite and purified via flash column chromatography using DCM and MeOH as eluents to give 1.60 g (72%) of the desired product as colorless oil.
Ή NMR (400 MHz, CDCI3) 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); 13C NMR (100 MHz, CDCI3) δ ppm; 70.1, 65.2, 62.9, 46.4, 31.9, 27.9, 26.0, 18.2, -4.36, -4.62; HRMS-ESI (m/z): [M+H]+ calcd for Ci3H32NO2Si: 262.2197, found 262.2197.
Préparation 2e: 4-[ierAButyl(dimethyI)silyl]oxy-5-morpholino-pentan-l-ol
Step A: [4-[tert-butyl(dimethyl)silyl]oxy-5-morpholino-pentyl] benzoate
1.50 g of Préparation 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 5 and (4-hydroxy-5-morpholino-pentyl) benzoate was obtained as a thick light yellow oil. LCMS-ESI (m/z): [M+H]+ calcd for C16H24NO4: 294.2, found 294.2.
The crude product was redissolved in 7 mL of Ν,Ν-dimethylformamide then 1.19 g of imidazole (17.52 mmol, 3 eq.) were added and after getting clear solution 1.76 g of tert-butyl-chlorodimethyl-silane (11.68 mmol, 2 eq.) was added in one portion. The mixture was stirred at 60°C 10 for 2 h. Cooled down to rt then EtOAc was added then the organic layer was washed with H2O, brine then dried over MgSÜ4, filtered then concentrated onto Celite. The crude product was purified via flash column chromatography using heptane and EtOAc as eluents to give 2.18 g (92%) of the desired product as colorless oil.
Ή NMR (400 MHz, DMSO-d0) δ ppm 7.96 (dd, 2H), 7.66 (t, 1H), 7.53 (t, 2H), 4.29 (t, 2H), 15 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); 13C NMR (100 MHz, CDCI3) 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]+ calcd for C22H38NO4S1: 408.3, found 408.3.
Step B: 4-[tert-butyl(dimethyl)silyl]oxy-5-morpholino-pentan-l-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 K2CO3 (16.06 mmol, 3.0 eq.) was added in one portion. After the addition the mixture left to stir at rt until full conversion was observed. Most of the volatiles was evaporated then EtOAc and water were added, separated and the organic phase was washed with brine, dried over MgSÜ4, filtered, concentrated. 1.49 g (72%) of the desired product as 25 colorless oil was obtained.
Ή NMR (400 MHz, DMSO-d6) 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, 4 H), 2.22 (ddd, 2H), 1.56-1.29 (m, 4H), 0.85 (s, 9H), 0.06 (s, 3H), 0.04 (s, 3H); 13C NMR (100 MHz, DMSO-d6) 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]+ calcd for 30 C15H34NO3S1: 304.2, found 304.3.
Préparation 2f: 4-[teri-Butyl(dimethyl)silyl]oxy-5-[2-(dimethylamino)ethyl-methyl20487 amiino]pentan-l-ol
Step A: [4-[tert-butyl(dimethyl)silyl]oxy-5-[2-(dimethylamino)ethyl-methyl-amino]pentyl] benzoate
908 mg of Préparation 2d, Step A (4.40 mmol, 1 eq.) and 1.35 g of Ν,Ν',Ν'-trimethylethane1,2-diamine (13.2 mmol, 3 eq.) were stirred in 12 mL of MeCN at 82°C for on. The reaction mixture was concentrated and [5-[2-(dimethylamino)ethyl-methyl-amino]-4-hydroxy-pentyl] benzoate was obtained as a thick light yellow oil. LC-MS-ESI (m/z): [M+H]+ calcd for C17H29N2O3: 309.2, found 309.3.
The crude product was redissolved in 4 mL Ν,Ν-dimethylformamide then 609 mg of imidazole (8.95 mmol, 3 eq.) were added and after getting clear solution 898 mg of tert-butyl-chlorodimethyl-silane (5.96 mmol, 2 eq.) was added in one portion. The mixture was stirred at 60°C for 2 h. Cooled down to rt then EtOAc was added then the organic layer was washed with H2O, brine then dried over MgSO4, filtered then concentrated onto Celite. The crude product was purified via flash column chromatography using DCM and MeOH as eluents to give 1.11 g (88%) of the desired product as colorless oil.
Ή NMR (400 MHz, DMSO-d6) 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); 13C NMR (100 MHz, DMSO-d6) 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, 17.8, -4.3, -4.8; LC-MS-ESI (m/z): [M+H]+ calcd for C23H43N2O3Si·. 423.3, found 423.3.
Step B: 4-[tert-butyl(dimethyl)silyl]oxy-5-[2-(dimethylamino)ethyl-methyl-amino]pentan-lol
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 the addition the mixture was stirred at rt until full conversion was observed. EtOAc and water were added, separated and the organic phase was washed with brine, dried over MgSO4, filtered, concentrated. 2.60 g (quant.) of the desired product as light yellow oil was obtained.
‘H NMR (500 MHz, DMSO-de) 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); 13C NMR (125 MHz, DMSO-d6) 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-ESI (m/z): [M+H]+ calcd for Ci6H39N2O2Si:
319.2775, found 319.2788.
Préparation 2g: 3-[tert-Butyl(dimethyI)silyl]oxy-4-morpholino-butan-l-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 CHCh then 4.97 5 g of 3-chlorobenzenecarboperoxoic acid (77w%, 28.80 mmol, 1.25 eq.) was added and stirred at rt for on. 60 mL of DCM was added then washed with cc. NaHCCh and brine, dried over MgSO4, filtered, concentrated and purified via flash column chromatography using heptane and EtOAc as eluents to give 3.81 g (86%) of the desired product as colorless oil.
Ή NMR (400 MHz, DMSO-îZ6) δ ppm 7.98 (d, 2H), 7.66 (t, 1H), 7.53 (t, 2H), 4.39 (t, 2H), 10 3.11-3.05 (m, 1H), 2.73 (t, 1H), 2.53 (dd, 1H), 2.03-1.82 (m, 2H); 13C NMR (100 MHz, DMSOdô) 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]+ calcd for C11H13O3: 193.1, found 193.1.
Step B: [3-[tert-butyl(dimethyl)silyl]oxy-4-morpholino-butyl] benzoate
723 mg of the product from Step A (3.76 mmol, 1 eq.) and 983 mg of morpholine (11.28 mmol, 15 3 eq.) were stirred in 10 mL MeCN at 82°C for 24 h. The reaction mixture was concentrated and (3-hydroxy-4-morpholino-butyl) benzoate was obtained as a thick light yellow oil. LC-MSESI (m/z): [M+H]+ calcd for Ci5H22NO4: 280.2, found 280.2.
The crude product was redissolved in 5 mL of Ν,Ν-dimethylformamide then 487 mg of imidazole (7.15 mmol, 3 eq.) were added and after getting clear solution 719 mg of tert-butyl20 chloro-dimethyl-silane (ATI mmol, 2 eq.) was added in one portion. The mixture was stirred at 60°C for 2 h. Cooled down to rt then EtOAc was added then the organic layer was washed with H2O, brine then dried over MgSO4, filtered then concentrated onto Celite. The crude product was purified via flash column chromatography using heptane and EtOAc as eluents to give 669 mg (71%) of the desired product as colorless oil.
’H NMR (400 MHz, DMSO-d6) 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); 13C NMR (100 MHz, DMSO-î/6) 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]+ calcd for C2iH36NO4Si: 394.2, found 394.3.
Step C: 3-[tert-butyl(dimethyl)silyl]oxy-4-morpholino-butan-l-ol
366 mg of the product from Step B (0.93 mmol, 1 eq) was dissolved in 2 mL of MeOH and 6.75 mL of THF then 186 mg of NaOH (4.65 mmol, 5.0 eq.) in 2.25 mL of water was added in one portion. After the addition the mixture left to stir at rt until full conversion was observed. EtOAc and water were added, separated and the organic phase was washed with brine, dried over MgSO4, filtered, concentrated. 262 mg (97%) of the desired product as colorless oil was 5 obtained.
Ή NMR (400 MHz, DMSO-tZ6) δ ppm 4.42 (br., 1H), 3.93-3.87 (m, 1H), 3.55 (t, 4H), 3.523.41 (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); 13C NMR (100 MHz, DMSO-rfy) 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]+ calcd for C14H32NO3S1: 290.2, found 10 290.3.
Préparation 2h: 3-[tert-Butyl(dimethyl)silyl]oxy-4-methoxy-butan-l-ol
Step A: methyl 3-hydroxy-4-methoxy-butanoate
To 1.00 g of methyl 4-methoxy-3-oxo-butanoate (6.84 mmol, 1 eq.) in 30 mL of MeOH was added 264 mg of NaBH4 (6.98 mmol, 1 eq.) in portions at 0 °C and the mixture was stirred at 15 0°C for 10 h. After concentration, the residue was diluted with water and extracted with EtOAc.
The combined organic phases were dried and concentrated to give 0.72 g (71%) of the desired product.
Ή NMR (400 MHz, DMSO-rfe) 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-methoxy-butanoate
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 was added 10 g of tert-butyl-chloro-dimethyl-silane (66.3 mmol, 1.1 eq.) in one portion and the mixture was stirred for 18 h. After dilution with 300 mL of brine and extraction with EtOAc, the combined organic phases were dried, concentrated, and purified 25 by flash column chromatography using heptane and EtOAc as eluents to give 11.77 g (75%) of the desired product.
Ή NMR (400 MHz, DMSO-ife) 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-methoxy-butan-l-ol
To 10.77 g of the product from Step B (41 mmol, 1 eq.) in 288 mL of DCM was slowly added 124 mL of DIBAL-H (1 M in THF, 124 mmol, 3 eq.) at 0 °C. After stirred for 2.5 h at 0 °C, 5 mL of water, 5 mL of a 15w% solution of NaOH, 0.5 mL of water and anhydrous MgSO4 were added consecutively. After 15 min of stirring, the mixture was filtered and concentrated to give 6.73 g (70%) of the desired product.
Ή NMR (500 MHz, DMSO-d6) 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); HRMSESI (m/z): [M+H]+ calcd for CnH27O3Si: 235.1729, found: 235.1725.
Préparation 2i: 4-[/erLButyl(dimethyl)silyl]oxy-3-methoxy-butan-l-ol
Step A: 3,4-dihydroxybutyl benzoate
The mixture of 20.1 g of but-3-enyl benzoate (113.5 mmol, 1 eq.), 21.8 g of 4-methyl-4-oxidomorpholin-4-ium;hydrate (161.3 mmol, 1.42 eq.), and 2.83 g of tetraoxoosmium (2.5 w% in 2methylpropan-2-ol, 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 h. After the reaction was quenched with the addition of 270 mL of a 1 M solution of Na2S2O3 at rt, the mixture was diluted with DCM and the organic phase was washed with water and brine, dried, and concentrated to give 22.22 g (93%) of 3,4dihydroxybutyl benzoate.
’H NMR (500 MHz, DMSO-î/6) 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); 13C NMR (125 MHz, DMSO-d6) δ ppm 166.2, 133.7, 129.5, 129.2, 68.5, 66.4, 62.5, 33.0; HRMS-ESI (m/z): [M+H]+ calcd for C11H15O4: 211.0970, found: 211.0971.
Step B: [4-[tert-butyl(dimethyl)silyl]oxy-3-hydroxy-butyl] 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 was added 7.17 g of tert-butyl-chloro-dimethyl-silane (47.6 mmol, 1 eq.) in one portion and the mixture was stirred for 1 h. After the mixture was diluted with EtOAc, the organic phase was washed with cc. NaHCO3, water and brine, dried over MgSO4, filtered, concentrated to give 14.1 g (91%) of the desired product.
JH NMR (400 MHz, DMSO-d6) 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-methoxy-butyl] benzoate
7.03 g of the product from Step B (21.7 mmol, 1 eq) and 13.9 g of N1,N1,N8,N8tetramethylnaphthalene-l,8-diamine (64.9 mmol, 3 eq.) were mixed in 420 mL of DCM then
8.0 g of trimethyloxonium;tetrafluoroborate (54.1 mmol, 2.5 eq.) was added in one portion and the mixture was stirred at rt for 18 h. Full conversion was observed. The mixture was filtered through a pad of Celite, washed with DCM then the organic phase was washed with water, brine, respectively. The solution was dried over MgSÛ4, filtered, concentrated onto Celite and purified via flash column chromatography using heptane and EtOAc as eluents to give 4.03 g (55%) of the desired product.
Ή NMR (400 MHz, DMSO-d6) 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-methoxy-butan-l-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 was added 5.1 g of K2CO3 (36.96 mmol, 3 eq.) at 0 °C and the mixture was stirred for 3 h at 0 °C and for 2 h at rt. After the addition of cc. NH4CI and DCM at 0 °C, the organic phase was washed with cc. NaHCO3, water and brine, dried over MgSÛ4, filtered, and concentrated to give 4.14 g (94%) of a 1:1 mixture of the desired product and methyl benzoate.
Ή NMR (400 MHz, DMSO-î/6) 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).
Préparation 2j: 3-[teri-Butyl(dimethyl)siIyl]oxy-2-methoxy-propan-l-ol
StepA: (2,2-dimethyl-l,3-dioxolan-4-yl)methylbenzoate
To 20 g of (2,2-dimethyl-l,3-dioxolan-4-yl)methanol (151 mmol, 1 eq.) and 25 mL of N,Ndiethylethanamine (182 mmol, 1.2 eq.) in 92 mL of DCM was added 21 mL of benzoyl chloride (182 mmol, 1.2 eq.) at 0 °C and the mixture was stirred at rt for 18 h. The reaction was quenched with cc. NaHCO3 and the organic phase was washed with brine, dried, and concentrated to give 37.0 g (98%) of the desired product.
3H NMR (400 MHz, DMSO-ώ) 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
The mixture of 20 g of the product from Step A (80 mmol, 1 eq.), 220 mL of a 1 N solution of HCl, and 220 mL of EtOH was stirred at rt for 18 h. After the reaction was quenched with cc. Na2CO3 and concentrated, the residue was extracted with EtOAc. The combined organic phases were washed with brine and dried to give 15.48 g (98.7%) of the desired product.
Ή NMR (400 MHz, DMSO-ifc) 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-hydroxy-propyl] 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 5 eq.) in 50 mL of DMF was added 4.04 g of tert-butyl-chloro-dimethyl-silane (26.8 mmol, 1 eq.) in one portion and the mixture was stirred for 1 h. After the mixture was diluted with EtOAc, the organic phase was washed with cc. NaHCOs, water and brine, dried over MgSO4, filtered, concentrated and purified by flash column chromatography using heptane and EtOAc as eluents to give 3.23 g (36%) of the desired product.
Ή NMR (400 MHz, DMSO-de) 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-methoxy-propyl] benzoate
3.0 g of the product from Step C (9.6 mmol, 1 eq) and 6.2 g of N1,N1,N8,N8tetramethylnaphthalene-l,8-diamine (28.9 mmol, 3 eq.) were mixed in 160 mL of DCM then 15 3.54 g of trimethyloxonium;tetrafluoroborate (23.9 mmol, 2.5 eq.) was added in one portion and the mixture was stirred at rt for 18 h. The mixture was filtered through a pad of Celite, washed with DCM then the organic phase was washed with water, brine, respectively. The solution was dried over MgSO4, filtered, concentrated onto Celite and purified via flash column chromatography using heptane and EtOAc as eluents to give 1.7 g (55%) of the desired product.
‘H NMR (400 MHz, DMSO-rfc) 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-methoxy-propan-l-ol
To 1.68 g of the product from StepD (5.17 mmol, 1 eq.) in 40 mL of a 1:1 mixture of THF and MeOH was added 2.19 g of K2CO3 (15.85 mmol, 3 eq.) at 0 °C and the mixture was stirred for 25 3 h at 0 °C and for 2 h at rt. After the addition of cc. NH4CI and DCM at 0 °C, the organic phase was washed with cc. NaHCOs, water and brine, dried over MgSÜ4, filtered, and concentrated to give 1.26 g (quant.) of a 1:0.25 mixture of the desired product and methyl benzoate.
Ή NMR (400 MHz, DMSO-cfc) 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).
Préparation 2k: 3-[terLButyl(dimethyl)silyl]oxy-2-morpholino-propan-l-ol
Step A: diethyl2-morpholinopropanedioate
To 16 g of morpholine (184 mmol, 1.6 eq.), 30 g of K2CO3 (217 mmol, 1.9 eq.) in 50 mL of MeCN was added 27 g of diethyl bromomalonate (113 mmol, 1 eq.) and the exotermic reaction was stirred for 0.5 h. After cooling the mixture to rt, it was filtered and concentrated and the 5 crude product was purified via flash column chromatography using heptane and EtOAc as eluents to give 26.2 g (94%) of the desired product.
NMR (400 MHz, DMSO-d6) 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-l,3-diol
To 3.0 g of L1AIH4 (79 mmol, 4.8 eq.) in 60 mL of THF was added 4.0 g of the product from Step A (16.3 mmol, 1 eq.) in 25 mL of THF at 0 °C and the mixture was stirred at 60 °C for 4 h. After cooling to rt, the mixture was treated with a 10 N solution of NaOH and the précipitation formed was filtered off and washed with DCM. After the phases were separated, the aqueous phase was extracted with DCM and the combined organic phases were washed 15 with brine, dried over MgSCU, filtered, and concentrated to give 1.4 g (53%) of the desired product.
Ή NMR (500 MHz, DMSO-î/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); 13C NMR (125 MHz, DMSO-d6) δ ppm 67.5, 67.3, 59.1, 50.5; IR: 3373, 2856; HRMS-EI (m/z): [M]+ calcd for C7H15NO3: 161.1052, found: 161.1053.
Step C: 3-[tert-Butyl(dimethyl) silyl]oxy-2-morpholino-propan-l -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 was added 1.24 g of tert-butyl-chloro-dimethyl-silane (8.23 mmo, 1 eq.) in 10 mL of DCM dropwise and the mixture was stirred at rt for 0.5 h. After the addition of DCM, the solution was washed with water and brine, dried, concentrated, and purified by flash 25 column chromatography using heptane and EtOAc as eluents to give 0.79 g (35%) of the desired product.
Ή NMR (500 MHz, DMSO-tfc) 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); 13C NMR (125 MHz, DMSO-d6) δ ppm 67.4, 67.4, 61.2, 59.2, 50.8, 26.3, 18.3, -5.0; IR: 2855; 30 HRMS-EI (m/z): [M-C4H9]+ calcd for C9H2oN03Si: 218.1217, found: 218.1202.
Préparation 3a: Methyl 5-[3-[4-[3-[teri-butoxycarbonyl(methyl)amino]prop-l-ynyl]2-fluoro-phenoxy]propyl]-2-[[5-[teri-butyl(dimethyl)silyl]oxy-4-[iertbutyl(diphenyl)silyl]oxy-pentyl]amino]thiazole-4-carboxylate
Step A : methyl 2-[tert-butoxycarbonyl-[5-[tert-butyl(dimethyl)silyl]oxy-4-[tertbutyl(diphenyl)silyl]oxy-pentyl]amino]-5-[3-[4-[3-[tert-butoxycarbonyl(methyl)amino]propl-ynyl]-2-fluoro-phenoxy]propyl]thiazole-4-carboxylate
Using Mitsunobu General Procedure starting from Préparation 1b as the appropriate carbamate and Préparation 2a as the appropriate alcohol, 2.5 g (61%) of the desired product was obtained.
Ή NMR (500 MHz, DMSO-d6) 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); 13C NMR (125 MHz, DMSO-d6) 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; HRMS-ESI (m/z): [M+H]+ calcd for C55H79FN3O9SSi2: 1032.5054, found 1032.5060.
Step B: methyl 5-[3-[4-[3-[tert-butoxycarbonyl(methyl)amino]prop-l-ynyl]-2-fluorophenoxy]propyl]-2-[[5-[tert-butyl(dimethyl)silyl]oxy-4-[tert-butyl(diphenyl)silyl]oxypentyl]amino]thiazole-4-carboxylate
Using Deprotection with HFIP General Procedure starting from the product from Step A as the appropriate carbamate, 1.2 g (53%) of the desired product was obtained.
Ή NMR (500 MHz, DMSO-d6) 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 (br., 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); 13C NMR (125 MHz, DMSO-dô) 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; HRMS-ESI (m/z): [M+H]+ calcd for C5oH7iFN307SSi2: 932.4530, found 932.4526.
Préparation 3b: Methyl 2-[ [5-[tert-butyI(dimethyl)siIyl]oxy-4-[tertbutyl(diphenyl)silyl]oxy-pentyl]amino]-5-[3-[4-[3-(dimethylamino)prop-l-ynyl]-2-fluorophenoxy]propyl]thiazo!e-4-carboxylate
Step A: methyl 2-[tert-butoxycarbonyl-[5-[tert-butyl(dimethyl)silyl]oxy-4-[tertbutyl(diphenyl)silyl]oxy-pentyl]amino]-5-[3-[4-[3-(dimethylamino)prop-l-ynyl]-2-fluorophenoxy]propyl]thiazole-4-carboxylate
Using Mitsunobu General Procedure starting from Préparation le as the appropriate carbamate and Préparation 2a as the appropriate alcohol, 3.2 g (65%) of the desired product was obtained.
Ή NMR (500 MHz, DMSO-d6) 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); 13C NMR (125 MHz, DMSO-d6) 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; HRMS-ESI (m/z): [M+H]+ calcd for C5iH73FN3O7SSi2: 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-l-ynyl]-2-fluoro-phenoxy]propyl]thiazole-4carboxylate
Using Deprotection with HFIP General Procedure starting from the product from Step A as the appropriate carbamate, 2.2 g (55%) of the desired product was obtained.
]H NMR (500 MHz, DMSO-d6) 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); 13C NMR (125 MHz, DMSO-d6) 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; HRMS-ESI (m/z): [M+H]+ calcd for C46H66FN3O5SSi2: 846.4162, found 846.4160.
Préparation 3c: Methyl 5-[3-[4-[3-[teri-butoxycarbonyl(methyl)amino]prop-l-ynyl]2-fluoro-phenoxy]propyl]-2-[4-[teri-butyl(dimethyl)siIyl]oxybutylamino]thiazole-4carboxylate
Step A: methyl 2-[tert-butoxycarbonyl-[4-[tert-butyl(dimethyl)silyl] oxybutyl]amino]-5-[3(2-fluoro-4-iodo-phenoxy)propyl]thiazole-4-carboxylate
Using Alkylation General Procedure starting from 5.36 g of Préparation la (10 mmol, 1 eq.) and 3.77 g of tert-butyl-(4ùodobiaoxy)-dimethyl-silane (3.10 mL, 12 mmol, 1.2 eq.) as the appropriate halide, 6.1 g (84%) of the desired product was obtained.
• ]H NMR (500 MHz, DMSO-d6) 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); 13C NMR (125 MHz, DMSO-dô) 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; HRMS-ESI (m/z): 5 [M+H]+ calcd for C29H45N2O6FS1SI: 723.1791, found 723.1775.
Step B: methyl 2-[4-[tert-butyl(dimethyl)silyl] oxybutylamino]-5-[3-(2-fluoro-4-iodophenoxy)propyl]thiazole-4-carboxylate
Using Deprotection with HFIP General Procedure starting from 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 10 obtained.
]H NMR (500 MHz, DMSO-dô) 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); 13C NMR (125 MHz, DMSO-dô) 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, 15 23.3, -4.8; HRMS-ESI (m/z): [M+H]+ calcd for C24H37FIN2O4SSi: 623.1266, found 623.1272.
Step C: methyl 5-[3-[4-[3-[tert-butoxycarbonyl(methyl)amino]prop-l-ynyl]-2-fluorophenoxy]propyl]-2-[4-[tert-butyl(dimethyl)silyl]oxybutylamino]thiazole-4-carboxylate
Using Sonogashira General Procedure starting from 3.00 g of the product from Step B (4.82 mmol, 1 eq.) and 1.63 g of tert-butylN-methyl-N-prop-2-ynyl-carbamate (9.63 mmol, 2 eq.) as 20 the appropriate acetylene, 2.50 g (65%) of the desired product was obtained.
*H NMR (500 MHz, DMSO-dô) 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); 13C NMR (125 MHz, DMSO-dô) δ ppm 129.1, 119.3, 115.4, 68.0, 62.7, 51.7, 44.3, 38.6, 33.8, 30.6, 30.1, 25 28.5, 26.3, 25.6, 23.3, -4.8; HRMS-ESI (m/z): [M+H]+ calcd for C33H5iFN3O6SSi: 664.3246, found 664.3245.
Préparation 3d: Methyl 2-[4-[terAbutyl(dimethyl)silyl]oxybutylamino]-5-[3-[4-[3(dimethylamino)prop-l-ynyl]-2-fluoro-phenoxy]propyl]thiazole-4-carboxylate
Using Sonogashira General Procedure starting from 3.00 g of Préparation 3c, Step B (4.82 30 mmol, 1 eq.) and 801 mg of N,N-dimethyl-N-prop-2-yn-l-amine (9.63 mmol, 2 eq.) as the appropriate acetylene, 2.20 g (79%) of the desired product was obtained.
φ Ή NMR (500 MHz, DMSO-d6) 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); 13C NMR (125 MHz, DMSO-d6) 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, 5 4.8; HRMS-ESI (m/z): [M+H]+ calcd for C29H45FN3O4SSi: 578.2878, found 578.2865.
Préparation 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 éthanol was cooled to 0°C, and then 7.46 g of ethyl 3-bromo-6-chloro-2-oxo-hexanoate (27.5 mmol, 1.1 eq.) was added dropwise at this température. After 15 min stirring at 0°C, 7 mL of TEA (5.06 g, 50 mmol, 2 eq.) was added. The resulting mixture was stirred overnight at rt. Full conversion was observed. The volatiles were removed in vacuo, then the résultant residue was portioned between EtOAc and water. The layers were separated then the organic layer was washed with water then followed with brine. The combined organic layers were dried over Na2SÜ4, filtered and the filtrate was concentrated under reduced pressure. Then it was purified via flash column chromatography using heptane and EtOAc as eluents to give 5 g (76%) of the desired product.
Ή NMR (400 MHz, DMSO-d6) 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); 13C NMR (125 MHz, DMSO-d6) δ ppm 165.6, 162.5, 137.4, 135.5, 60.5, 45.0, 34.1, 31.2, 24.4, 14.7; HRMS-ESI (m/z): [M+H]+ calcd for C10H16CIN2O2S: 263.0616, found 263.0615.
Préparation 3f: Methyl 5-[3-[4-[3-[tert-butoxycarbonyl(methyl)amino]prop-l-ynyl]2-fluoro-phenoxy] propyl] -2- [[5- [terf-butyl(dimethyl)sîlyl] oxy-4-methoxypentyl]amino]thiazole-4-carboxylate
Step A: methyl 2-[tert-butoxycarbonyl-[5-[tert-butyl(dimethyl)silyl]oxy-4-[tertbutyl(diphenyl)silyl]oxy-pentyl]amino]-5-[3-[4-[3-[tert-butoxycarbonyl(methyl)amino]prop25 1 -ynyl]-2-fluoro-phenoxy]propyl]thiazole-4-carboxylate
Using Mitsunobu General Procedure starting from 577 mg of Préparation 1b (1 mmol) as the appropriate carbamate and 496 mg of Préparation 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-l-ynyl]-2-fluoro30 phenoxy]propyl]-2-[[5-[tert-butyl(dimethyl)silyl]oxy-4-methoxy-pentyl]amino]thiazole-4carboxylate
Using Deprotection with HFIP General Procedure starting from 790 mg of the product from
Step A (0.95 mmol) as the appropriate carbamate, 270 mg (38%) of the desired product was obtained.
Ή NMR (500 MHz, DMSO-d6) 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); HRMS-ESI (m/z): [M+H]+ calcd for C35H55FN3O7SSi: 708.3508, found 708.3502.
Préparation 3g: Methyl 5-[3-[4-[3-(tert-butoxycarbonylamino)prop-l-ynyl]-2fluoro-phenoxy]propyl]-2-[4-[terAbutyl(dimethyl)silyl]oxybutylamino]thiazole-4carboxylate
Using Sonogashira General Procedure starting from 880 mg of Préparation 3c, Step B (1.41 mmol, 1 eq.) and 438 mg of tert-butyl N-prop-2-ynylcarbamate (2.82 mmol, 2 eq.) as the appropriate acetylene, 918 mg (85%) of the desired product was obtained.
Ή NMR (500 MHz, DMSO-d6) 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); 13C NMR (125 MHz, DMSO-dô) 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, 18.4, -4.8 HRMS-ESI (m/z): [M+H]+ calcd for C32H49FN3O6SSi: 650.3090, found 650.3093.
Préparation 3h: Methyl 5-[3-[4-[3-[terAbutoxycarbonyl(methyl)amino]prop-l-ynyl]2-fluoro-phenoxy]propyI]-2-[2-(2,2-dimethyl-l,3-dioxolan-4-yl)ethylamino]thiazole-4carboxylate
Step A: methyl 2-[tert-butoxycarbonyl-[2-(2,2-dimethyl-l,3-dioxolan-4-yl)ethyl]amino]-5[3-(2-fluoro-4-iodo-phenoxy)propyl]thiazole-4-carboxylate
Using Mitsunobu General Procedure starting from 2.68 g of Préparation la (5 mmol, 1 eq.) and 1.46 g of 2-(2,2-dimethyl-l,3-dioxolan-4-yl)ethanol (1.42 mL, 10 mmol, 2 eq.) as the appropriate alcohol, 2.8 g (84%) of the desired product was obtained.
1H NMR (500 MHz, DMSO-d6) δ ppm 7.57 (dd, 1H), 7.44 (dm, 1H), 6.96 (t, 1H), 4.12/4.02 (m+m, 2H), 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); 13C NMR (125 MHz,
DMSO-d6) 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; HRMS-ESI (m/z): [M+H]+ calcd for C26H35FIN2O7S: 665.1188, found 665.1175.
Step B: methyl 2-[2-(2,2-dimethyl-l,3-dioxolan-4-yl)ethylamino]-5-[3-(2-fluoro-4-iodophenoxy)propyl]thiazole-4-carboxylate
Using Deprotection with HFIP General Procedure starting from 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.
Ή NMR (500 MHz, DMSO-d6) 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, 10 2H), 1.99 (m, 2H), 1.76/1.72 (m+m, 2H), 1.31 (s, 3H), 1.25 (s, 3 H); HRMS-ESI (m/z): [M+H]+ calcd for C21H27FIN2O5S: 565.0663, found 565.0642.
Step C: methyl 5-[3-[4-[3-[tert-butoxycarbonyl(methyl)amino]prop-l-ynyl]-2-fluorophenoxy]propyl]-2-[2-(2,2-dimethyl-l,3-dioxolan-4-yl)ethylamino]thiazole-4-carboxylate
Using Sonogashira General Procedure starting from 400 mg of the product from Step B (0.71 15 mmol, 1 eq.) and 240 mg of tert-butyl N-methyl-N-prop-2-ynyl-carbamate (1.42 mmol, 2 eq.) as the appropriate acetylene, 300 mg (70%) of the desired product was obtained.
Ή NMR (500 MHz, DMSO-d6) 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, 20 3H); 13C NMR (125 MHz, DMSO-d6) δ ppm 164.5, 136.9, 136.4, 129.1, 119.3, 115.4, 85.2,
2.3, 73.8, 69.0, 68.0, 51.7, 41.4, 38.4, 33.8, 33.2, 30.6, 28.5, 27.3, 26.1, 23.3; HRMS-ESI (m/z): [M+H]+ calcd for C30H41FN3O7S: 606.2644, found 606.2650.
Préparation 3i: Methyl 5-[3-[4-[3-(dimethyl)amino]prop-l-ynyl]-2-fluorophenoxy]propyl]-2-[2-(2,2-dimethyl-l,3-dioxolan-4-yl)ethylamino]thiazole-4-carboxylate
Using Sonogashira General Procedure starting from 400 mg of Préparation 3h, Step B (0.71 mmol, 1 eq.) and 117 mg of N,N-dimethylprop-2-yn-l-amine (1.42 mmol, 2 eq.) as the appropriate acetylene, 250 mg (58%) of the desired product was obtained.
JH NMR (500 MHz, DMSO-d6) 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), 30 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); 13C NMR (125 MHz, DMSO-d6)ôppm 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; HRMSESI (m/z): [M+H]+ calcd for C26H35FN3O5S: 520.2281, found 520.2272.
Préparation 3i: Methyl 2-[2-(2,2-dimethyl-l,3-dioxolan-4-yl)ethylamino]-5-[3-[2fluoro-4- [3- [(4-methoxyphenyl)methyl-methyl-amino] prop-1- ynyl]phenoxy]propyl]thiazole-4-carboxylate
Using Sonogashira General Procedure starting from 400 mg of the product from Préparation 3h, Step B (0.71 mmol, 1 eq.) and 268 mg of N-[(4-methoxyphenyl)methyl]-N-methyl-prop-2yn-l-amine (1.42 mmol, 2 eq.) as the appropriate acetylene, 280 mg (63%) of the desired product was obtained.
]H NMR (500 MHz, DMSO-d6) δ ppm 7.61 (t, 1H), 7.33 (dd, 1H), 7.24 (dd, 1 H), 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); 13C 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, 15 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;
HRMS-ESI (m/z); [M+H]+ calcd for C33H41FN3O6S: 626.2694, found 626.2697.
Préparation 3k: Methyl 2-[2-(2,2-dimethyl-l,3-dioxolan-4-yl)ethylamino]-5-[3-[2fluoro-4-[3-[methyl(p-tolylsulfonyl)amino]prop-l-ynyl]phenoxy]propyl]thiazole-4carboxylate
Using Sonogashira General Procedure starting from 400 mg of Préparation 3h, Step B (0.71 mmol, 1 eq.) and 316 mg of N,4-dimethyl-N-prop-2-ynyl-benzenesulfonamide (1.42 mmol, 2 eq.) as the appropriate acetylene, 260 mg (55%) of the desired product was obtained.
]H NMR (500 MHz, DMSO-d6) 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, 25 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); 13C NMR (125 MHz, DMSO-d6) 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; HRMS-ESI (m/z): [M+H]+ calcd for C32H39FN3O7S2: 660.22079, found 660.2231.
Préparation 31: Methyl 5-[3-[4-[3-[tert-butoxycarbonyl(methyl)amino]prop-l-ynyl]20487
2-fluoro-phenoxy]propyl]-2-[[3-Rert-butyl(dimethyl)silyl]oxy-4-methoxybutyl] amino] thiazoIe-4-carboxylate
Step A: methyl2-[tert-butoxycarbonyl-[3-[tert-butyl(dimethyl)silyl]oxy-4-methoxybutyl]amino]-5-[3-[4-[3-[tert-butoxycarbonyl(methyl)amino]prop-l -ynyl]-2-fluoro5 phenoxy]propyl]thiazole-4-carboxylate
Using Mitsunobu General Procedure starting from 577 mg of Préparation 1b (1 mmol, 1 eq.) as the appropriate carbamate and 469 mg of Préparation 2h (2 mmol, 2 eq.) as the appropriate alcohol 794 mg (99%) of the desired product was obtained.
Step B: methyl 5-[3-[4-[3-[tert-butoxycarbonyl(methyl)amino]prop-l-ynyl]-2-fluoro10 phenoxy]propyl]-2-[[3-[tert-butyl(dimethyl)silyl]oxy-4-methoxy-butyl]amino]thiazole-4carboxylate
Using Deprotection with HFIP General 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.
Ή NMR (400 MHz, DMSO-d6) 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), 2.03-1.93 (m, 2H), 1.74-1.50 (m, 2H), 1.39 (s, 9H), 0.81 (s, 9H), 0.00 (s, 3H), -0.01 (s, 3H).
Préparation 3m: Methyl 5-[3-[4-[3-[terAbutoxycarbonyl(methyl)amino]prop-l-ynyI]20 2-fluoro-phenoxy] propyl] -2- [5- [tert-buty 1 (dimethy 1) sily 1] oxypentylamino] thiazole-4carboxylate
Step A: methyl2-[tert-butoxycarbonyl-[5-[tert-butyl(dimethyl)silyl]oxypentyl]amino]-5-[3(2-fluoro-4-iodo-phenoxy)propyl]thiazole-4-carboxylate
Using Alkylation General Procedure starting from 5.00 g of Préparation la (7.55 mmol, 1 25 eq.) and 2.97 g of tert-butyl-(5-iodopentoxy)-dimethyl-silane (9.06 mmol, 1.2 eq.) as the appropriate halide, 4.72 g (85%) of the desired product was obtained.
Ή NMR (500 MHz, DMSO-d6) 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); 13C NMR (125 MHz, DMSO-d6) δ ppm 30 163.1,156.3/153.2,151.8,147.0,143.2,135.6,133.9,124.9,117.6, 83.5, 82.3, 67.3, 62.5, 52.0,
6.5, 32.3, 30.5, 28.1, 27.6, 26.3, 22.9, 22.6, 18.1, -4.9; HRMS-ESI (m/z): [M+H]+ calcd for
C30H47FIN2O6SS1: 737.1947, found 737.1948.
Step_B: methyl 2-[[5-[tert-butyl(dimethyl)silyl]oxypentyl]amino]-5-[3-(2-fluoro-4-iodophenoxy)propyl]thiazole-4-carboxylate
Using Deprotection with HFIP General Procedure starting from 2.36 g of the product from 5 Step A (3.20 mmol) as the appropriate carbamate. After completion of the reaction the reaction mixture was evaporated to dryness under reduced pressure which resulted in the crude desired product.
Ή NMR (400 MHz, DMSO-d6) 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 10 (m, 2H), 0.84 (s, 9H), 0.00 (s, 6H).
Step C: methyl 5-[3-[4-[3-[tert-butoxycarbonyl(methyl)amino]prop-l-ynyl]-2-fluorophenoxy]propyl]-2-[5-[tert-butyl(dimethyl)silyl]oxypentylamino]thiazole-4-carboxylate
Using Sonogashira General Procedure starting from crude product from Step B and tert-butyl N-methyl-N-prop-2-ynyl-carbamate as the appropriate acetylene, 1.92 g (89% for 2 steps) of 15 the desired product was obtained.
‘H NMR (500 MHz, DMSO-d6) 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, 2 H), 2.86 (br., 3 H), 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); 13C NMR (125 MHz, DMSO-d6) δ ppm 129.1, 119.3, 115.4, 68.0, 62.8, 51.7, 44.5, 20 38.6, 33.8, 32.4, 30.6, 28.9, 28.5, 26.3, 23.3, 23.2, -4.9; HRMS-ESI (m/z): [M+H]+ calcd for
C34H53FN3O6SSi: 678.3403, found 678.3393.
Préparation 3n: Methyl 5-[3-[4-[3-[terf-butoxycarbonyl(methyl)amino]prop-l-ynyl]2-fluoro-phenoxy]propyl]-2-[3-[ierributyl(dimethyl)silyl]oxypropylamino]thiazole-4carboxylate
Step A: methyl 2-[tert-butoxycarbonyl-[3-[tert-butyl(dimethyl)silyl]oxypropyl]amino]-5-[3[4-[3-[tert-butoxycarbonyl(methyl)amino] prop-l-ynyl]-2-fluoro-phenoxy]propyl]thiazole-4carboxylate
Using Mitsunobu General Procedure starting from 577 mg of Préparation 1b (1 mmol, 1 eq.) as the appropriate carbamate and 380 mg oî3-[tert-butyl(dimethyl)silyl]oxypropan-l-ol (2 30 mmol, 2 eq.) as the appropriate alcohol, 600 mg (80%) of the desired product was obtained.
Step B: methyl 5-[3-[4-[3-[tert-butoxycarbonyl(methyl)amino]prop-1 -ynyl]-2-fluoro20487 phenoxy]propyl]-2-[3-[tert-butyl(dimethyl)silyl]oxypropylamino]thiazole-4-carboxylate
Using Deprotection with HFIP General Procedure starting from the product from Step A as the appropriate carbamate, 310 mg (47%) of the desired product was obtained.
Ή NMR (400 MHz, DMSO-d6) 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).
Préparation 3o: Methyl 5-[3-[4-[3-[terAbutoxycarbonyl(methyl)amino]prop-l-ynyl]2-fluoro-phenoxy]propyl]-2-[(2,2-dimethyl-l,3-dioxolan-4-yl)methylamino]thiazole-4carboxylate
Step A: methyl2-[tert-butoxycarbonyl-[(2,2-dimethyl-l,3-dioxolan-4-yl)methyl]amino]-5[3-[4-[3-[tert-butoxycarbonyl(methyl) amino]prop-1 -ynyl]-2-fluorophenoxy]propyl]thiazole-4-carboxylate
Using Miitsunobu General Procedure starting from 577 mg of Préparation 1b (1 mmol, 1 eq.) as the appropriate carbamate and 264 mg of (2,2-dimethyl-l,3-dioxolan-4-yl)methanol (2 mmol, 2 eq.) as the appropriate alcohol, 640 mg (92%) of the desired product was obtained.
Step B: methyl 5-[3-[4-[3-[tert-butoxycarbonyl(methyl)amino]prop-l-ynyl]-2-fluorophenoxy]propyl]-2-[(2,2-dimethyl-l,3-dioxolan-4-yl)methylamino]thiazole-4-carboxylate
Using Deprotection with HFIP General Procedure starting from the product from Step A as the appropriate carbamate, 282 mg (51%) of the desired product was obtained.
]H NMR (400 MHz, DMSO-d6) 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), 3.313.24 (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).
Préparation 3p: Methyl 5-[3-[4-[3-(dimethylamino)prop-l-ynyl]-2-fluorophenoxy]propyl]-2-[(2,2-dimethyl-l,3-dioxolan-4-yl)methylamino]thiazole-4-carboxylate
Step A: methyl 2-[tert-butoxycarbonyl-[(2,2-dimethyl-l,3-dioxolan-4-yl)methyl]amino]-5[3-[4-[3-(dimethylamino)prop-l-ynyl]-2-fluoro-phenoxy]propyl]thiazole-4-carboxylate
Using Mitsunobu General Procedure starting from 491 mg of Préparation le (1 mmol, 1 eq.) as the appropriate carbamate and 292 mg of (2,2-dimethyl-l,3-dioxolan-4-yl)methanol (2 mmol, 2 eq.) as the appropriate alcohol, 573 mg (92%) of the desired product was obtained.
S/ep B: Methyl 5-[3-[4-[3-(dimethylamino)prop-l-ynyl]-2-fluoro-phenoxy]propyl]-2-[(2,2dimethyl-l,3-dioxolan-4-yl)methylamino]thiazole-4-carboxylate
Using Deprotection with HFIP General Procedure starting from the product from Step A as the appropriate carbamate, 378 mg (74%) of Préparation 3p was obtained.
Ή NMR (400 MHz, DMSO-d6) 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), 3.333.26 (m, 1H), 3.12 (t, 2H), 2.23 (s, 6H), 2.05-1.96 (m, 2H), 1.34 (s, 3H), 1.26 (s, 3H).
Préparation 3q: Methyl 5-[3-[4-[3-[terLbutoxycarbonyl(methyl)amino]prop-l-ynyl]2-fluoro-phenoxy]propyl]-2-[[3-[tert-butyl(dimethyl)silyl]oxy-2-methoxypropyl]amino]thiazole-4-carboxylate
Step A: methyl2-[tert-butoxycarbonyl-[3-[tert-butyl(dimethyl)silyl]oxy-2-methoxypropyl]amino]-5-[3-[4-[3-[tert-butoxycarbonyl(methyl)amino]prop-l-ynyl]-2-fluorophenoxy]propyl]thiazole-4-carboxylate
Using Mitsunobu General Procedure starting from 577 mg of Préparation 1b (1 mmol, 1 eq.) as the appropriate carbamate and 440 mg of Préparation 2j (2 mmol, 2 eq.) as the appropriate alcohol, 780 mg (quant.) of the desired product was obtained.
Step B: methyl 5-[3-[4-[3-[tert-butoxycarbonyl(methyl)amino]prop-l-ynyl]-2-fluorophenoxy]propyl]-2-[[3-[tert-butyl(dimethyl)silyl]oxy-2-methoxy-propyl]amino]thiazole-4carboxylate
Using Deprotection with HFIP General Procedure starting from the product from Step A as the appropriate carbamate, 400 mg (58%) of the desired product was obtained.
Ή NMR (400 MHz, DMSO-d6) 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).
Préparation 3r: Methyl 5-[3-[4-[3-[terLbutoxycarbonyl(methyl)amino]prop-l-ynyl]2-fluoro-phenoxy]propyl]-2-[[3-[tert-butyl(dimethyl)silyl]oxy-2-morpholinopropy 1] amino] thiazole-4-carboxy late
Stei^A: methyl 2-[tert-butoxycarbonyl-[3-[tert-butyl(dimethyl)silyl]oxy-2-morpholinopropyl]amino]-5-[3-[4-[3-[tert-butoxycarbonyl(methyl) amino]prop-1 -ynyl]-2-fluorophenoxy]propyl]thiazole-4-carboxylate
Using Mitsunobu General Procedure starting from 577 mg of Préparation 1b (1 mmol, 1 eq.) as the appropriate carbamate and 413 mg of Préparation 2k (1.5 mmol, 1.5 eq.) as the appropriate alcohol, 473 mg (56%) of the desired product was obtained.
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-morpholino-propyl]amino]thiazole4-carboxylate
Using Deprotection with HFIP General Procedure starting from the product from Step A as the appropriate carbamate, 390 mg (93%) of the desired product was obtained.
Ή NMR (400 MHz, DMSO-d6) 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).
Préparation 3s: Methyl 5-[3-(2-fluoro-4-iodo-phenoxy)propyl]-2(methyIamino)thiazole-4-carboxylate
Step A: methyl 2-[tert-butoxycarbonyl(methyl)amino]-5-[3-(2-fluoro-4-iodophenoxy)propyl]thiazole-4-carboxylate
Using Alkylation General Procedure starting from 2.68 g of Préparation la (10 mmol, 1 eq.) and 1.70 g of iodomethane (12 mmol, 1.2 eq.) as the appropriate alkyl halide, 2.8 g (77%) of the desired product was obtained.
Step B: methyl 5-[3-(2-fluoro-4-iodo-phenoxy)propyl]-2-(methylamino)thiazole-4carboxylate
Using Deprotection with HFIP General Procedure starting from 2.0 g of the product from Step A (4.44 mmol) as the appropriate carbamate, 1.2 g (73%) of the desired product was obtained.
Ή NMR (500 MHz, DMSO-d6) 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); 13C NMR (125 MHz, DMSO20487 φ d6) δ ppm 165.4, 163.0, 147.0, 134.0, 124.9, 117.6, 68.0, 51.8, 31.1, 30.6, 23.3; HRMS-ESI (m/z): [M+H]+ calcd for C15H17FIN2O3S: 450.9983, found 450.9971.
Préparation 3t: Methyl 2-[5-[te^butyl(dimethyl)silyl]oxypentylamino]-5-[3-[4-[3(dimethylamino)prop-l-ynyl]-2-fluoro-phenoxy]propyl]thiazole-4-carboxylate
Using Sonogashira General Procedure starting from crude Préparation 3m, Step B and N,Ndimethylprop-2-yn-l-amine as the appropriate acetylene, 3.73 g (quant, for 2 steps) of the desired product was obtained.
HRMS-ESI (m/z): [M+H]+ calcd for C3oH47FN304SSi: 592.3035, found 592.3033.
Préparation 3u: Ethyl 2-amino-5-(3-{4-[3-(dimethylamino)prop-l-yn-l-yl]-210 fluorophenoxy}propyl)-l,3-thiazole-4-carboxylate
Step A: ethyl3-bromo-6-chloro-2-oxohexanoate
Bromine (1.6 mL, 31.2 mmol, 1.2 eq) was added to a stirred solution of ethyl 6-chloro-2oxohexanoate (5 g, 26.0 mmol, 1 eq) in chloroform (70 mL). The reaction was stirred at ambient température for 6.5 h then diluted with dichloromethane and washed with 10% aqueous sodium 15 thiosulphate followed by water then brine. The organic extract was dried (magnésium sulfate) and the solvent removed in vacuo to afford the desired compound (7 g, 25.8 mmol, 99%) as a colourless oil.
'H NMR (400 MHz, DMSO-d6) 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 20 (t, J = 7.1 Hz, 3H).
Step B: ethyl 5-(3-chloropropyl)-2-acetamido-l,3-thiazole-4-carboxylate
A-Acetylthiourea, (3.05 g, 25.78 mmol, 1 eq) was added to a stirred solution of the product from Step A (7 g, 25.8 mmol, 1 eq) in éthanol (150 mL). The reaction was heated at reflux for 6 h, then allowed to cool to ambient température and the solvent was removed in vacuo. The 25 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 température for 2 h the mixture was diluted with dichloromethane then washed with water (x2) followed by brine. The organic extract was dried (magnésium sulfate) then the solvent removed in vacuo. The crude material was triturated with diethyl ether and the solid 30 was collected by filtration and washed with diethyl ether to yield the desired product (1.1 g, 3.78 mmol, 13.9%). Two cycles of filtrate évaporation, re-trituration, and filtration afforded further product batches (3.42 g, 11.76 g, 43%). Finally, évaporation of the final filtrate and purification by automated flash column (CombiFlash Rf, 12 g RediSep™ silica cartridge) eluting in a gradient of 0 - 100% ethyl acetate in Ao-heptane afforded further desired product (1.46 g, 5.02 mmol, 18.5%) [overall yield: 76%].
LC/MS (C11H15CIN2O3S) 291 [M+H]+; RT 1.08 (LCMS-V-B1);
Ή NMR (400 MHz, DMSO-d6) 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)-l,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, 10 0.12 mol, 6 eq) in acetonitrile (130 mL) was heated at reflux for 6 h. The mixture was cooled to ambient température and the solvent removed in vacuo. The mixture was diluted with ethyl acetate and washed with water and brine. The organic extract was dried (magnésium sulfate) and concentrated in vacuo. The résultant solid was triturated in diethyl ether then collected by filtration affording the desired product as a cream solid (6.28 g, 16.4 mmol, 82 %).
LC/MS (C11H15IN2O3S) 383 [M+H]+; RT 0.96 (LCMS-V-B1)
Ή NMR (400 MHz, DMSO-d6) 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: ethyl5-(3-{4-[3-(dimethylamino)prop-l-yn-l-yl]-2-fluorophenoxy}propyl)-2acetatnido-l,3-thiazole-4-carboxylate
Sodium hydride (60% in minerai oil) (0.56 g, 14.1 mmol, 2 eq) was added to a stirred solution of the product from Step C (1.64 g, 8.46 mmol, 1.2 eq) in dimethylformamide (20 mL). The reaction was stirred at ambient température for 30 min then cooled in an ice bath and a solution of the product from Préparation 6b (2.7 g, 7.05 mmol, 1 eq) in dimethylformamide (20 mL) was added slowly. After 40 min the reaction was allowed to warm to ambient température and stirred 25 for 5 h. The solvent was removed in vacuo, and the crude material was partitioned between ethyl acetate and water, washed with brine, and dried (magnésium sulfate). The solvent was removed in vacuo and purification by automated flash column chromatography (CombiFlash Rf, 80 g RediSep™ silica cartridge) eluting in a gradient of 0 - 10% methanol in dichloromethane afforded the desired product as a cream solid (1.48 g, 3.32 mmol, 47%).
LC/MS (C22H26FN3O4S) 448 [M+H]+; RT 0.76 (LCMS-V-B1) φ Ή NMR (400 MHz, DMSO-d6) δ 12.41 (s, 1H), 7.30 (dd, J = 11.9, 2.3 Hz, 1H), 7.25 - 7.18 (m, 1H), 7.17 - 7.10 (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-l-yn-1 -yl]-2-fluorophenoxy}propyl)-
1,3- thiazole-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 éthanol (40 mL) was heated at 60 °C for 18 h. The reaction was allowed to cool to ambient température then the solvent removed in vacuo. The 10 résultant 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 then eluted with 3.5 N methanolic ammonia. The solvent was removed in vacuo and the résultant oil was purified by automated flash column chromatography (CombiFlash Rf, 80 g RediSep™ silica cartridge) eluting in a gradient of 0 — 10% methanol in dichloromethane to afford the desired product as 15 a cream solid (0.78 g, 1.93 mmol, 58%).
LC/MS (C20H24FN3O3S) 406 [M+H]+; RT 0.706 (LCMS-V-B1)
Ή NMR (400 MHz, DMSO-d6) 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).
Préparation 3v: Ethyl 2-amino-5-(3-phenoxypropyl)-l,3-thiazole-4-carboxylate
Step A: ethyl5-bromo-2-acetamido-l,3-thiazole-4-carboxylate
A solution of ethyl 2-amino-5-bromo-l,3-thiazole-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 température for 3 days. The reaction 25 was diluted with dichloromethane, washed successively with water and brine, then dried (magnésium sulfate) and the solvent removed in vacuo. The résultant solid was triturated with diethyl ether, filtered, and dried under vacuum to afford the desired product as an off-white solid (1.8 g, 6.14 mmol, 77%).
LC/MS (C8H9BrN2O3S) 293 [M+H]+; RT 1.01 (LCMS-V-B1)
Hi NMR (400 MHz, DMSO-d6) 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-l-yn-1 -yl)-1,3-thiazole-4-ca.rboxylate
The product from Step A (200 mg, 0.68 mmol, 1 eq), phenyl propargyl ether (0.18 mL, 1.36 mmol, 2 eq), triethylamine (0.28 mL, 2.05 mmol, 3 eq), copper (I) 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 sparged with nitrogen (10 min) 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 (magnésium sulfate). The solvent was removed in vacuo and purification by automated flash column chromatography (CombiFlash Rf, 12 g RediSep™ silica cartridge) eluting with a gradient of 0 - 100% ethyl acetate in Ao-heptane afforded the desired product as an off-white solid (122 mg, 0.35 mmol, 52%).
LC/MS (C17H16N2O4S) 345 [M+H]+; RT 1.21 (LCMS-V-B1)
Hl NMR (400 MHz, DMSO-d6) 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)-l,3-thiazole-4-carboxylate
Ethyl acetate (10 mL) was added to a flask containing the product from Step B (122 mg, 0.35 mmol, 1 eq) and platinum(IV) oxide (8.04 mg, 0.04 mmol, 0.1 eq) under a nitrogen atmosphère. The mixture was shaken at ambient température under an atmosphère of hydrogen for 6.5h. Further platinum(IV) oxide (spatula tip) was added and the reaction was shaken under a hydrogen atmosphère for a further 18 h. The mixture was filtered through a methanol-wet pad of celite, eluted with methanol, and the solvent removed in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 12 g RediSep™ silica cartridge) eluting with a gradient of 0 - 100% ethyl acetate in Ao-heptane afforded the desired product as a cream solid (77.2 mg, 0.22 mmol, 63%).
LC/MS (C17H20N2O4S) 349 [M+H]+; RT 1.22 (LCMS-V-B1) *H NMR (400 MHz, DMSO-d6) 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).
Step D: ethyl 2-amino-5-(3-phenoxypropyl)-l,3-thiazole-4-carboxylate φ Hydrochloric acid (4M 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 éthanol (5 mL). The reaction was heated at 60 C for 5 h. The solvent was removed in vacuo then the residue was dissolved in methanol and loaded onto a methanol-wet SCX-2 cartridge (5g). The cartridge was washed 5 with methanol, then eluted with 3.5N methanolic ammonia, and the solvent was removed in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 12 g RediSep™ silica cartridge) eluting with a gradient of 0 - 10% methanol in dichloromethane afforded the desired product as a yellow gum (48.2 mg, 0.16 mmol, 71%).
LC/MS (Ci5Hi8N2O3S) 307 [M+H]+; RT 1.16 (LCMS-V-B1)
Ή NMR (400 MHz, DMSO-d6) 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).
Préparation 3w: Ethyl 2-amino-5-(3-methoxypropyI)-l,3-thiazole-4-carboxylate
Step A: ethyl2-acetamido-5-(3-methoxyprop-l-yn-1 -yl)-l,3-thiazole-4-carboxylate
Dimethylformamide (10 mL) was added to the product from Préparation 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 (I) iodide (26 mg, 0.14 mmol, 0.1 eq) then tetrakis(triphenylphosphine)palladium(0) (78.8 mg, 0.07 mmol, 0.05 eq) was added. The vessel was sparged with nitrogen (10 min) then heated at 100°C for 1 h under microwave irradiation.
The solvent was removed in vacuo and purification by automated flash column chromatography (CombiFlash Rf, 12 g RediSep™ silica cartridge) eluting with a gradient of 0 - 5% methanol in dichloromethane afforded a solid that was triturated with diethyl ether to afford the desired product as a cream solid (213.7 mg, 0.76 mmol, 55%). The filtrate was evaporated to afford a second batch of desired product as an orange gum (94.6 mg, 0.34 mmol, 24%) [overall yield
79%].
LC/MS (Ci2Hi4N2O4S) 283 [M+H]+; RT 1.00 (LCMS-V-B1) ‘H NMR (400 MHz, DMSO-d6) 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 2:1 ethyl acetate / methanol (15 mL) was added to a flask containing catalytic platinum(IV) oxide under a nitrogen atmosphère. The vessel was evacuated and back-filled with nitrogen (x3), then evacuated and placed under an atmosphère of hydrogen. After shaking at ambient température for 18 h, the reaction was filtered through celite, eluted with methanol, and the solvent removed in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 12 g RediSep™ silica cartridge) eluting with a gradient of 0 — 100% ethyl acetate in Ao-heptane afforded the desired product as a cream solid (207 mg, 0.72 mmol, 66%).
LC/MS (C12H18N2O4S) 287 [M+H]+; RT 0.99 (LCMS-V-B1)
Ή NMR (400 MHz, DMSO-d6) 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 (4M 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 éthanol (10 mL). The mixture was heated to 60 °C for 7 h then allowed to cool to ambient température and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 12 g RediSep™ silica cartridge) eluting with a gradient of 0 - 10% methanol in dichloromethane afforded the desired product as a brown gum (219 mg, 0.81 mmol, 97%).
LC/MS (C10H16N2O3S) 245 [M+H]+; RT 0.88 (LCMS-V-B1) *H NMR (400 MHz, DMSO-d6) 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).
Préparation 3x: Ethyl 2-amino-5-[3-(2-fluorophenoxy)propyl]-l,3-thiazole-4carlboxylate
Step A: ethyl2-acetamido-5-(3-hydroxyprop-l-yn-l-yl)-l,3-thiazole-4-carboxylate
To a mixture of the product from Préparation 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 (I) iodide (65 mg, 0.34 mmol, 0.1 eq) in dimethylformamide (20 mL) was added tetrakis(triphenylphosphine)palladium(0) (197 mg, 0.17 mmol, 0.05 eq). The mixture was sparged with nitrogen (10 min) then heated at 100 °C for 1 h under microwave irradiation. The solvent was removed in vacuo then purification by automated flash column chromatography (CombiFlash Rf, 40 g RediSep™ silica cartridge) eluting with a gradient of 0 - 10% methanol in dichloromethane afforded the desired product as a brown solid (1.01 g, 3.75 mmol, > 100%).
φ LC/MS (C11H12N2O4S) 269 [M+H]+; RT 0.82 (LCMS-V-B1)
Ή NMR (400 MHz, DMSO-d6) δ 12.80 (s, 1H), 5.45 (t, J = 6.0 Hz, 1H), 4.37 (d, J = 6.1 Hz, 2H), 4.27 (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-hydroxypropyl)-l,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 added to a flask containing platinum(IV) oxide hydrate (spatula tip) under a nitrogen atmosphère. The reaction was evacuated and back-filled with nitrogen (x3), then evacuated and placed under an atmosphère of hydrogen. The reaction was shaken at ambient température for 30 h, then filtered through celite, eluted with methanol, and the solvent removed 10 in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 24 g RediSep™ silica cartridge) eluting with a gradient of 0 - 10% methanol in dichloromethane afforded the desired product as a brown gum (582 mg, 2.14 mmol, 57%).
LC/MS (C11H16N2O4S) 273 [M+H]+; RT 0.81 (LCMS-V-B1)
Ή NMR (400 MHz, DMSO-d6) δ 12.39 (s, 1H), 4.54 (t, J = 5.1 Hz, 1H), 4.25 (q, J = 7.1 Hz, 15 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]-l,3-thiazole-4-carboxylate
A solution of 2-fluorophenol (0.1 mL, 1.1 mmol, 1.5 eq), the product from Step B (200 mg, 0.73 mmol, 1 eq) and triphenylphosphine (289 mg, 1.1 mmol, 1.5 eq) in tetrahydrofuran (10 20 mL) was cooled in an ice bath then diisopropyl azobicarboxylate (0.22 mL, 1.1 mmol, 1.5 eq) was added dropwise. The reaction was allowed to warm gradually to ambient température and then stirred for 18 h. The mixture was diluted with ethyl acetate, washed with brine, and the organic phase was dried (magnésium sulfate) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 24 g RediSep™ silica cartridge) 25 eluting with a gradient of 0 - 100% ethyl acetate in heptane afforded the desired product as a white solid (162 mg, 0.44 mmol, 60%).
LC/MS (Ci7Hi9FN2O4S) 367 [M+H]+; RT 1.22 (LCMS-V-B1) 1H NMR (400 MHz, DMSO-d6) 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), 30 2.18 - 2.03 (m, 2H), 1.27 (t, J = 7.1 Hz, 3H).
Step D: ethyl 2-amino-5-[3-(2-fluorophenoxy)propyl]-l,3-thiazole-4-carboxylate
A solution of the product from Step C (217 mg, 0.59 mmol, 1 eq) and hydrochloric acid (4M in dioxane; 1.48 mL, 5.92 mmol, 10 eq) in éthanol (8 mL) was heated at 60 °C ovemight. The mixture was allowed to cool to ambient température then the solvent removed in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 12 g RediSep™ silica cartridge) eluting with a gradient of 0 - 10% methanol in dichloromethane afforded the desired product as a yellow foam (181 mg, 0.56 mmol, 94%).
LC/MS (C15H17FN2O3S) 325 [M+H]+; RT 1.16 (LCMS-V-B1)
Ή NMR (400 MHz, DMSO-d6) 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).
Préparation 3y: Ethyl 2-amino-5-[3-(4-bromo-2-fluorophenoxy)propyI]-l,3thiazoIe-4-carboxylate
Step A: ethyl5-[3-(4-bromo-2-fluorophenoxy)propyl]-2-acetamido-l,3-thiazole-4carboxylate
A solution of the product from Préparation 3x, Step B (248 mg, 0.91 mmol, 1 eq), 4-bromo-2fluorophenol (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 diisopropylazodicarboxylate (0.27 mL, 1.37 mmol, 1.5 eq) was added dropwise. The reaction was stirred in the ice-bath for 30 min then stirred at ambient température for 3 h. The reaction was diluted with ethyl acetate and washed with water then brine. The organics were dried (magnésium sulfate) and the solvent removed in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 24 g RediSep™ silica cartridge) eluting with a gradient of 0 - 100% ethyl acetate in heptane afforded the desired product as a white solid (229 mg, 0.51 mmol, 56%).
LC/MS (Ci7Hi8BrFN2O4S) 445 [M+H]+; RT 1.36 (LCMS-V-B1)
Ή NMR (400 MHz, DMSO-d6) 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]-l,3-thiazole-4-carboxylate
A solution of the product from Step A (229 mg, 0.51 mmol, 1 eq) and hydrochloric acid (4M m dioxane; 1.29 mL, 5.14 mmol, 10 eq) in éthanol (8 mL) was heated at 60 °C for 18 h. The reaction was allowed to cool to ambient température then the solvent was removed in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 12 g RediSep™ silica cartridge) eluting with a gradient of 0 - 10% methanol in dichloromethane afforded the desired product as a yellow gum (199 mg, 0.49 mmol, 96%).
LC/MS (Ci5Hi6BrFN2O3S) 405 [M+H]+; RT 1.30 (LCMS-V-B1)
Ή NMR (400 MHz, DMSO-d6) 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, J = 7.1 Hz, 3H).
Préparation 3z; Ethyl 2-[(2-methoxyethyl)amino]-l,3-thiazole-4-carboxylate
Ethyl bromopyruvate (2.5 mL, 17.9 mmol, 1.07 eq) was added to a suspension of 2methoxyethylthiourea (2.25 g, 16.8 mmol, 1 eq) in éthanol (40 mL) at 0 °C under a nitrogen atmosphère and the mixture stirred for 1 h at ambient température. Triethylamine (4.7 mL, 33.5 mmol, 2 eq) was added and the mixture heated at 75 °C for 18 h. The solution was allowed to cool to ambient température and was partitioned between ethyl acetate (300 mL) and water (200 mL). The organic phase was washed with water (200 mL) and brine, dried (magnésium sulfate) and concentrated in vacuo. Purification by flash column chromatography (100 g silica) eluting with 1:1 ethyl acetate / heptane gave a solid that was triturated with heptane (20 mL), filtered, washed with heptane (2 x 15 mL) and dried under vacuum to afford the desired product as a pale brown solid (3.15 g, 13.7 mmol, 82%).
LC/MS (C9H14N2O3S) 231 [M+H]+; RT 0.88 (LCMS-V-B1)
Ή NMR (400 MHz, CDCI3) 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).
Préparation 3za: Ethyl 5-[4-(benzyloxy)piperidin-l-yl]-2-(methylamino)-l,3-thiazole4-carboxylate
Step A: ethyl5-[4-(benzyloxy)piperidin-l-yl]-l,3-thiazole-4-carboxylate
A solution of ethyl 5-bromothiazole-4-carboxylate (500 mg, 2.12 mmol, 1 eq), 4-benzyloxypiperidine hydrochloride (579 mg, 2.54 mmol, 1.2 eq) and l,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 was allowed to cool to ambient température then concentrated in vacuo, partitioned between dichloromethane and water, and the organic phase washed with brine, dried (magnésium sulfate) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 24 g RediSep™ silica cartridge) eluting with a gradient of 0 — 100% ethyl acetate in Ao-heptane afforded the desired product as a yellow oil (499 mg, 1.44 mmol, 68%).
LC/MS (C18H22N2O3S) 347 [M+H]+; RT 1.08 (LCMS-V-B1)
Ή .NMR (400 MHz, DMSO-d6) 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-l-yl]-2-bromo-l,3-thiazole-4-carboxylate
A-Bromosuccinimide (308 mg, 1.73 mmol, 1.2 eq) was added to a stirred solution of the product frorn Step A (499 mg, 1.44 mmol, 1 eq) in acetonitrile (20 mL) and the mixture was stirred at ambient température overnight. The reaction was diluted with ethyl acetate, washed with water followed by brine, dried (magnésium sulfate), and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 24 g RediSep™ silica cartridge) eluting with a gradient of 0 — 60% ethyl acetate in zso-heptane afforded the desired product as a yellow solid (454 mg, 1.07 mmol, 74%).
LC/MS (Ci8H2iBrN2O3S) 427 [M+H]+; RT 1.26 (LCMS-V-B1)
Ή NMR (400 MHz, DMSO-d6) 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-l-yl]-2-{[(tert-butoxy)carbonyl](methyl)amino}-l,3thiazole-4-carboxylate
The product from Step B (454 mg, 1.07 mmol, 1 eq) and ieri-butyl methylcarbamate (0.19 mL, 1.28 mmol, 1.2 eq) were added 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) under a nitrogen atmosphère. Césium carbonate (522 mg, 1.6 mmol, 1.5 eq) was added and the mixture heated at 100 °C overnight. The reaction was allowed to cool to ambient température and concentrated in vacuo. The residue was dissolved in ethyl acetate, then washed with water followed by brine, dried (magnésium sulfate) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 50 g RediSep™ silica cartridge) eluting with a gradient of 0 - 50% ethyl acetate in Ao-heptane afforded the desired product as a cream solid (339 mg, 0.71 mmol, 67%).
LC/MS (C24H33N3O5S) 476 [M+H]+; RT 1.37 (LCMS-V-B1) XH NMR (400 MHz, DMSO-d6) 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-l-yl]-2-(methylamino)-l,3-thiazole-4-carboxylate
Trifluoroacetic acid (0.53 mL, 7.11 mmol, 10 eq) was added to a stirred solution of the product from Step C (339 mg, 0.71 mmol, 1 eq) in dichloromethane (10 mL) at 0 °C and the mixture was allowed to warm to ambient température and stir overnight. The reaction was partitioned between dichloromethane and saturated aqueous sodium bicarbonate, and the organic phase washed with brine, dried (magnésium sulfate), and concentrated in vacuo. The material was triturated with diethyl ether, filtered, and dried under vacuum to afford the desired product as a white solid (213 mg, 0.57 mmol, 80%).
LC/MS (C19H25N3O3S) 376 [M+H]+; RT 1.07 (LCMS-V-B1)
Ή NMR (400 MHz, DMSO-d6) 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).
Préparation 3zb: Ethyl 5-{l-[(ieri-butoxy)carbonyl]azetidin-3-yl}-2-(methylamino)l,3-thiazole-4-carboxylate
Step A: ({!-[(tert-butoxy)carbonyljazetidin-3-yl}methyl)(iodo)zinc
To an oven-dried Schlenk flask was added zinc (3.3 g, 50.5 mmol, 1.5 eq) and the vessel was heated at 160 °C for 20 min under vacuum then allowed to cool to ambient température and placed under a nitrogen atmosphère. Dimethylacetamide (25 mL) was added followed by slow addition of a mixture of trimethylchlorosilane (0.69 mL, 5.42 mmol, 0.16 eq) and 1,2dibromoethane (0.49 mL, 5.69 mmol, 0.17 eq). The résultant mixture was allowed to cool to ambient température then a solution of l-boc-3-(iodomethyl)azetidine (10 g, 33.65 mmol, 1 eq) in dimethylacetamide (7 mL) was added slowly and the mixture was stirred overnight. Cannulation through a filter (cotton-wool/ celite / cotton-wool) into a dry 25 mL Schlenk tube afforded the desired product as a 0.62M solution (as determined by titration with a 0.5M solution of iodine) that was used without further characterisation.
Step B: tert-butyl 3-(3-ethoxy-2,3-dioxopropyl)azetidine-l-carboxylate
A solution of copper(I) iodide (3.54 g, 18.6 mmol, 2 eq) in tetrahydrofuran (20 mL) was cooled to -20 °C then the product from Step A (0.62M in dimethylacetamide; 30 mL, 18.6 mmol, 2 eq) was added slowly and the mixture was stirred at -20 °C for 10 min then at 0 °C for 20 min. 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 température and stir for 1 h. The reaction was quenched by addition of saturated aqueous ammonium chloride, extracted with ethyl acetate (x2), and the combined organic extracts were washed with brine, dried (magnésium sulfate), and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 40 g RediSep™ silica cartridge) eluting with a gradient of 0 - 30% ethyl acetate in Ao-heptane afforded the desired product as a colourless oil (0.57 g, 2.1 mmol, 23%).
Ή 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-[(lZ)-3-ethoxy-3-oxo-2-[(trimethylsilyl)oxy]prop-l-en-l-yl]azetidine-lcarboxylate
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) was slowly added triethylamine (0.38 mL, 2.73 mmol, 1.3 eq) and the mixture was stirred at ambient température for 3.5 h. The reaction was partitioned between ethyl acetate and water, and the organic phase was washed with brine, dried (magnésium sulfate) and concentrated in vacuo to afford the desired product as a colourless oil (643 mg, 1.87 mmol, 89%).
Ή 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-(l-bromo-3-ethoxy-2,3-dioxopropyl)azetidine-l-carboxylate
A solution of the product from Step C (643 mg, 1.87 mmol, 1 eq) and A-bromosuccinimide (367 mg, 2.06 mmol, 1.1 eq) in tetrahydrofuran (15 mL) was heated at reflux for 2.5 h then allowed to cool to ambient température. The reaction was partitioned between ethyl acetate and saturated aqueous sodium bicarbonate and the organic phase was washed with brine, dried (magnésium sulfate), and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 24 g RediSep™ silica cartridge) eluting with a gradient of 0 - 40% ethyl acetate in ώο-heptane afforded the desired product as a yellow gum (507 mg, 1.45 mmol, 77%).
*Η 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)-l,3-thiazole-4-carboxylate
A solution of the product from Step D (452 mg, 1.29 mmol, 1 eq) and A-methylthiourea (140 mg, 1.55 mmol, 1.2 eq) in éthanol (8 mL) was heated at reflux for 4.5 h then allowed to cool to ambient température. The résultant precipitate was collected by filtration, washed with éthanol (2 mL), and dried under vacuum to afford the desired product as a white solid (161 mg, 0.5 mmol, 39%). The filtrate was transferred to a microwave vial and hydrochloric acid (4M in 1,4dioxane, 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 3.5N methanolic ammonia. The solvent was removed in vacua and purification by automated flash column chromatography (CombiFlash Rf, 24 g RediSep™ silica cartridge) eluting with a gradient of 0 - 15% 2.33N methanolic ammonia in dichloromethane afforded the desired product as a colourless gum (115 mg, 0.48 mmol, 37%). The two product batches were combined and used in the subséquent steps.
LC/MS (C10H15N3O2S) 242 [M+H]+; RT 0.34 (LCMS-V-B1)
Ή NMR (400 MHz, DMSO-d6) 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-{l-[(tertbutoxy)carbonyl]azetidin-3-yl}-l,3-thiazole-4-carboxylate
To a solution of the product of Step E (298 mg, 1.23 mmol, 1 eq) in dichloromethane (10 mL) was added di-teri-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 température overnight. The reaction was partitioned between dichloromethane and water, and the organic phase was washed with brine, dried (magnésium sulfate), and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 24 g RediSep™ silica cartridge) eluting with a gradient of 0 - 40% ethyl acetate in isoheptane afforded the desired product as a white foam (390 mg, 0.88 mmol, 72%).
LC/MS (C20H31N3O6S) 442 [M+H]+; RT 1.47 (LCMS-V-B1) ‘H NMR (400 MHz, DMSO-d6) 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-{l-[(tert-butoxy)carbonyl]azetidin-3-yl}-2-(methylamino)-l,3-thiazole-4carboxylate
A solution of the product from Step F (390 mg, 0.88 mmol, 1 eq) in 1,1,1,3,3,3-hexafluoro-2propanol (3 mL) was heated at 100 °C for 55 min under microwave irradiation. The mixture was concentrated in vacuo and purification by automated flash column chromatography (CombiFlash Rf, 12 g RediSep™ silica cartridge) eluting with a gradient of 0 — 100% ethyl acetate in Ao-heptane afforded the desired product as a white solid (199 mg, 0.58 mmol, 66%).
LC/MS (C15H23N3O4S) 342 [M+H]+; RT 0.98 (LCMS-V-B1)
Ή NMR (400 MHz, DMSO-d6) 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).
Préparation 3zc: Methyl 5-(3-{4-[3-(dimethylamino)prop-l-yn-l-yl]-2fluorophenoxy}-2,2-dimethylpropyl)-2-(methylamino)-l,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, was added sulfuric acid (2 mL) and the mixture was stirred at ambient température overnight. The reaction was concentrated in vacuo then quenched by portionwise addition of saturated aqueous sodium bicarbonate (200 mL) and washed with Ao-heptane (x2). The aqueous phase was cooled to 0 °C, acidified with 6N aqueous hydrochloric acid, and extracted with diethyl ether (x2). The combined organic extracts were dried (magnésium sulfate), and concentrated in vacuo to afford the desired product as a clear oil (13.7 g, 85.4 mmol, 62%).
Ή 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 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 h, then warmed to 0 °C and stirred at this température for 45 min. The reaction was quenched by the slow addition of methanol (50 mL), then concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 220 g RediSep™ silica cartridge) eluting with a gradient of 0 - 50% ethyl acetate in Ao-heptane afforded the desired product as a clear oil (8.14 g, 55.7 mmol, 65%).
XH NMR (400 MHz, DMSO-d6) 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) was added terAbutyl(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 température overnight. The reaction 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 (magnésium sulfate) and purification by automated flash column chromatography (CombiFlash Rf, 330 g RediSep™ silica cartridge) eluting with a gradient of 0 - 10% ethyl acetate in Ao-heptane afforded the desired product as a clear oil (14.4 g, 37.4 mmol, 67%).
XH NMR (400 MHz, DMSO-d6) 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-l-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 diisobutyl aluminum hydride (IM in tetrahydrofuran; 112 mL, 112 mmol, 3 eq) was added and the mixture was stirred at -78 °C for 2 h. Saturated aqueous ammonium chloride was added and the mixture stirred at -78 °C for 10 min, then allowed to warm to ambient température. The mixture was filtered through a pad of celite eluting with ethyl acetate, and the organics were dried (magnésium sulfate) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 220 g RediSep™ silica cartridge) eluting with a gradient of 0 - 15% ethyl acetate in zso-heptane afforded the desired product as a clear oil (12.5 g, 35 mmol, 94%).
LC/MS (C22H32O2Si) 357 [M+H]+; RT 2.68 (LCMS-V-C)
Ή NMR (400 MHz, DMSO-d6) δ 7.65 - 7.56 (m, 4H), 7.53 - 7.40 (m, 6H), 4.28 (t, J = 5.0 5 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 10 methanesulfonyl chloride (3.25 mL, 42 mmol, 1.2 eq) and the mixture was stirred at ambient température overnight. The reaction was diluted with dichloromethane, washed with saturated aqueous sodium bicarbonate, dried (PTFE phase separator), and concentrated in vacuo to afford the desired product as a yellow oil (15.1 g, 34.8 mmol, 99%).
Ή NMR (400 MHz, DMSO-d6) δ 7.66 - 7.58 (m, 4H), 7.53 - 7.40 (m, 6H), 4.27 (t, J = 7.4 15 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) was added lithium bromide (9.07 g, 105 mmol, 3 eq) portionwise and the mixture stirred at 70 °C for 4 h, then allowed to cool to ambient température. The reaction was concentrated in 20 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 cartridge) eluting with a gradient of 0 - 2% ethyl acetate in Ao-heptane afforded the desired product as a clear oil (12.8 g, 30.5 mmol, 88%).
Ή NMR (400 MHz, DMSO-d6) 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
To an oven-dried flask was added magnésium (496 mg, 20.4 mmol, 1.2 eq) and this was stirred vigorously under nitrogen for 20 min. A solution of the product from Step F (7.13 g, 17 mmol, 30 1 eq) in tetrahydrofuran (12 mL) was sparged with nitrogen (10 min) and a portion of this solution was added to the magnésium (without stirring). The mixture was briefly heated to reflux (heat-gun) and a crystal of iodine was added. Stirring was initiated, and the remaining solution was added to the magnésium suspension at such a rate as to maintain gentle reflux. After complété addition the mixture was heated at reflux for 2 h, then allowed to cool to ambient température. A stirred solution of diethyl oxalate (2.31 mL, 17 mmol, 1 eq) in tetrahydrofuran (6 mL) was sparged with nitrogen (10 min) then cooled to -78 °C. To this was added the above Grignard reagent solution portionwise, then the mixture was allowed to gradually warm to ambient température over 2 h. The reaction partitioned between ethyl acetate and saturated aqueous ammonium chloride, and the organic phase was dried (magnésium sulfate) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 220 g RediSep™ silica cartridge) eluting with a gradient of 0 - 8% ethyl acetate in isoheptane afforded the desired product as a clear oil (3.06 g, 6.95 mmol, 41%).
*H 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).
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) was added portionwise a solution of the product from Step G (3.06 g, 6.94 mmol, 1 eq) in chloroform (40 mL) and the reaction was heated at reflux for 16 h. The reaction mixture was allowed to cool to ambient température then filtered through a pad of celite, eluted with dichloromethane, and evaporated. Purification by automated flash column chromatography (CombiFlash Rf, 80 g RediSep™ silica cartridge) eluting with a gradient of 0 - 10% ethyl acetate in zso-heptane afforded the desired product as a clear oil (2.62 g, 5.04 mmol, 72%).
LC/MS (C26H35BrO4Si) 441 [M-Ph]+; RT 1.36 (LCMS-V-B2)
Ή 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: ethyl5-{3-[(tertbutyldiphenylsilyl)oxy]-2,2-dimethylpropyl}-2-(methylamino)-l,3thiazole-4-carboxylate
To a solution of the product from Step GH (2.62 g, 5.04 mmol, 1 eq) in éthanol (35 mL) was added A-methylthiourea (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 h, then allowed to cool to ambient température.
The reaction 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 cartridge) eluting with a gradient of 0 - 50% ethyl acetate in zTo-heptane afforded the desired product as a yellow solid (1.45 g, 2.84 mmol, 56%).
LC/MS (C28H38N2O3S1S) 511 [M+H]+; RT 1.28 (LCMS-V-B2)
Ή NMR (400 MHz, DMSO-d6) 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}-l,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 di-terz-butyl dicarbonate (0.84 mL, 3.92 mmol, 2 eq) and the mixture was stirred at ambient température for 3 h. The reaction was partitioned between ethyl acetate and water, and the organic phase was dried (magnésium sulfate), and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 24 g RediSep™ silica cartridge) eluting with a gradient of 0 - 15% ethyl acetate in No-heptane afforded the desired product as a yellow oil (1.19 g, 1.95 mmol, 100%).
LC/MS (C33H46N2O5SiS) 611 [M+H]+; RT 1.48 (LCMS-V-B2) XH NMR (400 MHz, DMSO-d6) 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)-l,3thiazole-4-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 (IM in tetrahydrofuran; 2.95 mL, 2.95 mmol, 3 eq) and the mixture was stirred at ambient température for 2 h. The reaction was partitioned between ethyl acetate and water, and the organic phase was dried (magnésium sulfate) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 12 g RediSep™ silica cartridge) eluting with a gradient of 0 - 100% ethyl acetate in Ao-heptane afforded the desired product as a white solid (178 mg, 0.52 mmol, 53%).
LC/MS (Ci5H24N2O5S) 345 [M+H]+; RT 1.87 (LCMS-V-C) ‘H NMR (400 MHz, DMSO-d6) 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)l,3-thiazole-4-carboxylate
To a solution of the product from Step J (178 mg, 0.52 mmol, 1 eq) in 1:1 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 température overnight. The reaction 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 cartridge) eluting with a gradient of 0 50% ethyl acetate in Ao-heptane afforded the desired product as a white solid (119 mg, 0.33 mmol, 64%).
LC/MS (C16H26N2O5S) 359 [M+H]+; RT 2.10 (LCMS-V-C) XH NMR (400 MHz, DMSO-d6) 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)2,2-dimethylpropyl]-l,3-thiazole-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-4-iodophenol (158 mg, 0.66 mmol, 2 eq), triphenylphosphine (174 mg, 0.66 mmol, 2 eq) and diisopropylazodicarboxylate (131 pL, 0.66 mmol, 2 eq) and the mixture was heated at 120 °C for 24 h. The reaction 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 cartridge) eluting with a gradient of 0 - 20% ethyl acetate in Ao-heptane afforded the desired product as a clear gum (136 mg, 0.24 mmol, 71%).
LC/MS (C22H28FIN2O5S) 579 [M+H]+; RT 2.76 (LCMS-V-C)
Ή NMR (400 MHz, DMSO-d6) 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)propl-yn-l-yl]-2-fluorophenoxy}-2,2-dimethylpropyl)-l,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-l-propyne (38.1 pL, 0.35 mmol, 1.5 eq), copper (I) iodide (4.48 mg, 0.02 mmol, 0.1 eq) and N,N-diisopropylethylamine (81.9 pL, 0.47 mmol, 2 eq). The mixture was sparged 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 was partitioned between ethyl acetate and brine, and the organic phase was dried (magnésium sulfate) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 4 g RediSep™ silica cartridge) eluting with a gradient of 0 - 4% methanol in dichloromethane afforded the desired product as a brown gum (27 mg, 0.05 mmol, 22%).
LC/MS (C27H36FN3O5S) 534 [M+H]+; RT 2.053 (LCMS-V-C)
Ή NMR (400 MHz, DMSO-d6) 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-l-yn-l-yl]-2-fluorophenoxy}-2,2dimethylpropyl)-2-(methylamino)-l,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 température for 2 h. The reaction was diluted with dichloromethane, then cooled to 0 °C and neutralised by the addition of 2N aqueous sodium hydroxide. 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 cartridge) eluting with a gradient of 0 7% methanol in dichloromethane afforded the desired product as a brown solid (8 mg, 0.02 mmol, 37%).
LC/MS (C22H28FN3O3S) 434 [M+H]+; RT 1.62 (LCMS-V-C)
Ή NMR (400 MHz, DMSO-d6) 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).
Préparation 3zd: Methyl 5-{[l-({4-[3-(dimethylamino)prop-l-yn-l-yl]-2fluorophenoxy}methyl)cyclopropyl]methyl}-2-(methylamino)-l,3-thiazole-4-carboxylate
Step A: 2-(l-{[(tert-butyldiphenylsilyl)oxy]methyl}cyclopropyl)acetonitrile
To a solution of 2-[l-(hydroxymethyl)cyclopropyl]acetonitrile (8 g, 72 mmol, 1 eq) in 5 dimethylformamide (200 mL) was added teri-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 température overnight. The reaction was partitioned between iso-heptane (600 mL) and 2:1 water / saturated aqueous ammonium chloride (600 mL) and the aqueous phase was extracted with iso-heptane (600 mL). The combined organic extracts were dried (magnésium sulfate) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 330 g RediSep™ silica cartridge) eluting with a gradient of 0 - 6% ethyl acetate in isoheptane afforded the desired product as a clear oil (23 g, 65.7 mmol, 91%).
‘H NMR (400 MHz, DMSO-d6) 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-(l-{[(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, was added diisobutyl aluminum hydride (IM in hexanes; 72.3 mL, 72.3 mmol, 1.1 eq) portionwise and the mixture was stirred at 0 °C for 1.5 h. The reaction was quenched by the addition of water, then 2N aqueous hydrochloric acid (100 mL) was added, followed by 20 saturated aqueous potasium sodium tartrate (500 mL). The mixture was extracted with dichloromethane (2 x 400 mL), and the combined organic extracts were dried (magnésium sulfate) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 330 g RediSep™ silica cartridge) eluting with a gradient of 0 - 10% ethyl acetate in iso-heptane afforded the desired product as a clear oil (19.6 g, 55.5 mmol, 85%).
LC/MS (C22H28O2S1) 353 [M+H]+; RT 1.19 (LCMS-V-B2) rH NMR (400 MHz, DMSO-d6) 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-(l-{[(tert-butyldiphenylsilyl)oxy]methyl}cyclopropyl)ethan-l-ol
To a solution of the product of Step B (19.6 g, 55.5 mmol, 1 eq) in methanol (200 mL), cooled to 0 °C, was added sodium borohydride (2.31 g, 61.1 mmol, 1.1 eq) portionwise and the mixture was stirred at ambient température for 3 h. The reaction was concentrated in vacuo, then partitioned between ethyl acetate and water, and the organic phase was dried (magnésium sulfate) and concentrated in vacuo to afford the desired product as a clear oil (18.8 g, 53 mmol, 96%).
'11 NMR (400 MHz, DMSO-d6) 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 D: 2-(l-{[(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 température overnight. The reaction 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 afford the desired product as a brown oil (23.3 g, 53 mmol, 100%).
*H NMR (400 MHz, DMSO-d6) 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 température and the solvent removed in vacuo. The residue was partitioned between dichloromethane and brine, and the organic phase was washed with 10% aqueous sodium thiosulfate, dried (magnésium sulfate), and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 330 g RediSep™ silica cartridge) eluting with a gradient of 0 - 2% diethyl ether in zso-heptane afforded the desired product as a clear oil (17.4 g, 41.6 mmol, 77%).
Ή NMR (400 MHz, DMSO-d6) 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-(l-{[(tert-butyldiphenylsilyl)oxy]methyl}cyclopropyl)-2-oxobutanoate
To an oven-dried flask was added magnésium (1.21 g, 49.9 mmol, 1.2 eq) and this was vigorously stirred 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 sparged with nitrogen (10 min) then an aliquot of this solution was added to the magnésium (without stirring), the mixture was briefly heated at reflux and a crystal of iodine was added. Stirring was initiated and the remaining solution was added to the magnésium at such a rate as to maintain gentle reflux. After complété addition the mixture was heated at reflux for 2 h, then allowed to cool to ambient température. A solution of diethyl oxalate (5.64 mL, 41.6 mmol, 1 eq) in tetrahydrofuran (15 mL) was sparged with nitrogen (10 min) then cooled to -78 °C. The above Grignard reagent solution was slowly added then the mixture was allowed to warm to ambient température over 2 h. The reaction was partitioned between ethyl acetate and saturated aqueous ammonium chloride, and the organic phase was dried (magnésium sulfate) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 330 g RediSep™ silica cartridge) eluting with a gradient of 0 - 8% ethyl acetate in zso-heptane afforded the desired product as a clear oil (10 g, 22.8 mmol, 55%).
Ή NMR (400 MHz, DMSO-d6) 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-(l-{[(tert-butyldiphenylsilyl)oxy]methyl}cyclopropyl)-2oxobutanoate
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 was filtered through celite, eluted with dichloromethane, and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 220 g RediSep™ silica cartridge) eluting with a gradient of 0 - 8% ethyl acetate in Ao-heptane afforded the desired product as a yellow oil (2.09 g, 4.04 mmol, 18%).
Ή NMR (400 MHz, DMSO-d6) 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-[(l-{[(tert-butyldiphenylsilyl)oxy]methyl}cyclopropyl)methyl]-2(methylamino)-l,3-thiazole-4-carboxylate
To a solution of the product from Step G (2.09 g, 4.04 mmol, 1 eq) in éthanol (30 mL) was added TV-methylthiourea (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 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 cartridge) eluting with a gradient of 0 — 50% ethyl acetate in isoheptane afforded the desired product as an orange solid (1.8 g, 3.53 mmol, 88%).
LC/MS (C28H36N2O3SiS) 509 [M+H]+; RT 2.69 (LCMS-V-C)
Ή NMR (400 MHz, DMSO-d6) 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 ·{[(tertbutyldiphenylsilyl)oxy]methyl}cyclopropyl)methyl]-l,3-thiazole-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-ieri-butyl dicarbonate (1.51 mL, 7.08 mmol, 2 eq) and the mixture was stirred at ambient température overnight. The reaction was partitioned between ethyl acetate and water, and the organic phase was dried (magnésium sulfate) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 40 g RediSep™ silica cartridge) eluting with a gradient of 0 - 15% ethyl acetate in zso-heptane afforded the desired product as a yellow oil (2.03 g, 3.33 mmol, 94%).
LC/MS (C33H44N2O5SiS) 609 [M+H]+; RT 1.98 (LCMS-V-C) XH NMR (400 MHz, DMSO-d6) 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}-l,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 (IM in tetrahydrofuran; 10 mL, 10 mmol, 3 eq) and the mixture was stirred at 70 °C for 2.5 h. The reaction was partitioned between ethyl acetate and water, and the organic phase was dried (magnésium sulfate) and concentrated in vacuo. Purification by automated flash column chiOmatography (CombiFlash Rf, 24 g RediSep™ silica cartridge) eluting with a gradient of 0 - 100% ethyl acetate in zso-heptane afforded the desired product as a yellow solid (460 mg, 1.34 mmol, 40%).
LC/MS (C15H22N2O5S) 343 [M+H]+; RT 1.77 (LCMS-V-C)
Ή NMR (400 MHz, DMSO-d6) 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-{[l(hydroxymethyl)cyclopropyl]methyl}-l,3-thiazole-4-carboxylate
To a solution of the product from Step J (460 mg, 1.34 mmol, 1 eq) in 1:1 dichloromethane / methanol (12 mL), cooled to 0 °C, was added (trimethylsilyl)diazomethane (2M in hexanes; 0.87 mL 1.75 mmol, 1.3 eq) and the mixture was stirred at ambient température overnight. The reaction 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 cartridge) eluting with a gradient of 0 50% ethyl acetate in zso-heptane afforded the desired product as a white solid (418 mg, 1.17 mmol, 87%).
LC/MS (C16H24N2O5S) 357 [M+H]+; RT 1.98 (LCMS-V-C)
Ή NMR (400 MHz, DMSO-d6) 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-({l-[(2-fluoro-4iodophenoxy)methyl]cyclopropyl}methyl)-l,3-thiazole-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-4-iodophenol (558 mg, 2.35 mmol, 2 eq), triphenylphosphine (615 mg, 2.35 mmol, 2 eq) and diisopropylazodicarboxylate (0.46 mL, 2.35 mmol, 2 eq) and the mixture was heated at 120 °C overnight, then allowed to cool to ambient température. The reaction 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, 12 g RediSep™ silica cartridge) eluting with a gradient of 0
100
- 20% ethyl acetate in /so-heptane afforded the desired product as a yellow oil (649 mg, 1.13 mmol, 96%).
LC/MS (C22H26FIN2O5S) 577 [M+H]+; RT 2.65 (LCMS-V-C) *H NMR (400 MHz, DMSO-d6) 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-{[!-({4-[3(dimethylamino)prop-l-yn-l-yl]-2-fluorophenoxy}methyl)cyclopropyl]methyl}-I,3-thiazole4-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-l-propyne (182 pL, 1.69 mmol, 1.5 eq), copper(I) iodide (21.4 mg, 0.11 mmol, 0.1 eq) and Λζ/V-diisopropylethylamine (392 pL, 2.25 mmol, 2 eq). The mixture was sparged 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 was partitioned between ethyl acetate and brine, and the organic phase was dried (magnésium sulfate) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 24 g RediSep™ silica cartridge) eluting with a gradient of 0 - 7% methanol in dichloromethane afforded the desired product as a brown solid (173 mg, 0.33 mmol, 29%).
LC/MS (C27H34FN3O5S) 532 [M+H]+; RT 2.004 (LCMS-V-C)
Ή NMR (400 MHz, DMSO-d6) 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-{[l-({4-[3-(dimethylamino)prop-l-yn-l-yl]-2fluorophenoxy}methyl)cyclopropyl]methyl}-2-(methylamino)-l,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 température for 2 h. The reaction was diluted with dichloromethane, then cooled to 0 °C and neutralised by the addition of 2N aqueous sodium hydroxide. 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 cartridge) eluting with a gradient of 0 20487
101
7% methanol in dichloromethane afforded the desired product as a yellow solid (84 mg, 0.19 mmol, 60%).
LC/MS (C22H26FN3O3S) 432 [M+H]+; RT 1.55 (LCMS-V-C)
Ή NMR (400 MHz, DMSO-d6) 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).
Préparation 3ze: Ethyl 2-{|4-(/e/7dmtoxy)-4-oxobutyl|amino}-l ,3-thiazole-4carboxylate
Step A: ethyl2-{[4-(tert-butoxy)-4-oxobutyl][(tert-butoxy)carbonyl]amino}-l,3-thiazole-4carboxylate
To a solution of ethyl 2-[(ierAbutoxycarbonyl)amino]-l,3-thiazole-4-carboxylate (5.2 g, 19.1 mmol, 1 eq) and zm-butyl 4-hydroxybutanoate (4.59 g, 28.6 mmol, 1.5 eq) in tetrahydrofuran (150 mL) was added triphenylphosphine (7.51 g, 28.6 mmol, 1.5 eq), followed by dropwise addition of diisopropylazodicarboxylate (5.64 mL, 28.6 mmol, 1.5 eq) and the mixture was stirred at ambient température for 1 h. The reaction was partitioned between ethyl acetate and water, the aqueous phase was extracted with ethyl acetate (3 x 60 mL), and the combined organic extracts were washed with brine (100 mL), dried (magnésium sulfate), and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 120 g RediSep™ silica cartridge) eluting with a gradient of 0 - 20% ethyl acetate in zso-heptane afforded the desired product as a colourless oil (7.7 g, 18.6 mmol, 97%).
LC/MS (Ci9H3oN206S) 415 [M+H]+; RT 1.49 (LCMS-V-B1)
Ή NMR (400 MHz, DMSO-d6) 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}-l,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-2propanol (40 mL) was heated in a sealed flask at 80 °C for 7 h. The reaction was concentrated in vacuo and purification by automated flash column chromatography (CombiFlash Rf, 80 g RediSep™ silica cartridge) eluting with a gradient of 0 - 35% ethyl acetate in zso-heptane afforded the desired product as a white solid (3.61 g, 11.5 mmol, 62%).
LC/MS (C14H22N2O4S) 315 [M+H]+; RT 1.20 (LCMS-V-B1)
102 φ JH NMR (400 MHz, DMSO-d6) 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).
Préparation 3zf: Ethyl 5-[(2S)-3-{4-[3-(dimethylamino)prop-l-yn-l-yl]-25 fluorophenoxy}-2-methylpropyl]-2-(methylamino)-l,3-thiazole-4-carboxylate
Step A: [(2R)-3-bromo-2-methylpropoxy](tert-butyl)diphenylsilane
To a solution of (7?)-(-)-3-bromo-2-methyl-l-propanol (5 g, 32.68 mmol, 1 eq) in dimethylformamide (120 mL) was added imidazole (4.32 mL, 65.4 mmol, 2 eq) and tertbutyl(chloro)diphenylsilane (11.1 mL, 42.5 mmol, 1.3 eq) and the mixture was stirred at 10 ambient température overnight. The reaction was partitioned between zso-heptane (300 mL) and 2:1 water / saturated aqueous ammonium chloride (200 mL), the aqueous phase was extracted with zso-heptane (200 mL), and the combined organic extracts were dried (magnésium sulfate) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 220 g RediSep™ silica cartridge) eluting with a gradient of 0 — 2% diethyl 15 ether in zxo-heptane afforded the desired product as a clear oil (12.1 g, 30.9 mmol, 95%).
Ή NMR (400 MHz, DMSO-d6) 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 dimethylsulfoxide (120 20 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 was partitioned between diethyl ether and water and the organic phase was dried (magnésium sulfate) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 220 g RediSep™ silica cartridge) eluting with a gradient of 0 - 3% ethyl acetate in zso-heptane afforded the desired product as a clear oil (7.17 25 g, 21.2 mmol, 69%).
’II NMR (400 MHz, DMSO-d6) 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: (3S)-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 30 mL), cooled to 0 °C, was added diisobutylaluminum hydride (23.4 mL, 23.4 mmol, 1.1 eq) and the mixture was stirred overnight. Water was added, followed by 2N aqueous hydrochloric acid
102 φ ΧΗ NMR (400 MHz, DMSO-d6) 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).
Préparation 3zf: Ethyl 5-[(2S)-3-{4-[3-(dimethylamino)prop-l-yn-l-yl]-25 fluorophenoxy}-2-methylpropyI]-2-(methylamino)-l,3-thiazole-4-carboxylate
Step A: [(2R)-3-bromo-2-methylpropoxy](tert-butyl)diphenylsilane
To a solution of (R)-(-)-3-bromo-2-methyl-l-propanol (5 g, 32.68 mmol, 1 eq) in dimethylformamide (120 mL) was added imidazole (4.32 mL, 65.4 mmol, 2 eq) and tertbutyl(chloro)diphenylsilane (11.1 mL, 42.5 mmol, 1.3 eq) and the mixture was stirred at 10 ambient température overnight. The reaction was partitioned between zso-heptane (300 mL) and 2:1 water / saturated aqueous ammonium chloride (200 mL), the aqueous phase was extracted with zso-heptane (200 mL), and the combined organic extracts were dried (magnésium sulfate) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 220 g RediSep™ silica cartridge) eluting with a gradient of 0 - 2% diethyl 15 ether in zso-heptane afforded the desired product as a clear oil (12.1 g, 30.9 mmol, 95%).
Ή NMR (400 MHz, DMSO-d6) 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 dimethylsulfoxide (120 20 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 was partitioned between diethyl ether and water and the organic phase was dried (magnésium sulfate) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 220 g RediSep™ silica cartridge) eluting with a gradient of 0 - 3% ethyl acetate in zso-heptane afforded the desired product as a clear oil (7.17 25 g, 21.2 mmol, 69%).
Ή NMR (400 MHz, DMSO-d6) 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: (3S)-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 30 mL), cooled to 0 °C, was added diisobutylaluminum hydride (23.4 mL, 23.4 mmol, 1.1 eq) and the mixture was stirred overnight. Water was added, followed by 2N aqueous hydrochloric acid
103
B (36 mL) and saturated aqueous potassium sodium tartrate (180 mL). The mixture was diluted with further water and dichloromethane and the organic phase 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 cartridge) eluting with 5 a gradient of 0 - 6% ethyl acetate in zw+heptanc afforded the desired product as a clear oil (6.21 g, 18.2 mmol, 86%).
XH NMR (400 MHz, DMSO-d6) 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).
StepD: (3S)-4-[(tert-butyldiphenylsilyl)oxy]-3-methylbutan-l-ol
To a solution of the product of Step C (6.21 g, 18.24 mmol, 1 eq) in methanol (80 mL), cooled in an ice-water bath, was added sodium borohydride (759 mg, 20.1 mmol, 1.1 eq) and the mixture was allowed to warm to ambient température and stir for 3 h. The reaction was concentrated in vacuo, then partitioned between ethyl acetate and water, and the organic phase 15 was dried (magnésium sulfate) and concentrated in vacuo to afford the desired product as a clear oil (6.36 g, 18.6 mmol, > 100%).
* H NMR (400 MHz, DMSO-d6) 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 of 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 température overnight. The reaction was partitioned between dichloromethane and saturated aqueous sodium 25 bicarbonate, and the organic phase was separated and dried (PTFE phase separator) and concentrated in vacuo to afford the desired product as a brown oil (8.28 g, 19.7 mmol, > 100%).
> H NMR (400 MHz, DMSO-d6) 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
104
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 5h. The reaction 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 cartridge) eluting with a gradient of 0 - 2% diethyl ether in Ao-heptane afforded the desired product as a clear oil (6.3 g, 15.5 mmol, 79%).
* H NMR (400 MHz, DMSO-d6) δ 7.66 - 7.58 (m, 4H), 7.53 - 7.40 (m, 6H), 3.64 - 3.47 (m, 4H), 2.08 - 1.93 (m, 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
To an oven-dried flask under nitrogen was added magnésium (453 mg, 18.7 mmol, 1.2 eq) and the solid was vigorously stirred for 20 min then 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 the magnésium followed by iodine (39.4 mg, 0.16 mmol, 0.01 eq) and the mixture was briefly heated at reflux. The remaining solution was added at such a rate as to maintain a gentle reflux, and upon complété addition the mixture was heated for 2 h at reflux then allowed to cool to ambient température. To a stirred solution of diethyl oxalate (2.11 mL, 15.5 mmol, 1 eq) in tetrahydrofuran (6 mL), cooled to -78 °C, was added the Grignard solution via cannula, and the mixture was allowed to warm to ambient température and stir for 5 h. The reaction was partitioned between ethyl acetate and saturated aqueous ammonium chloride, and the organic phase was dried (magnésium sulfate), and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 80 g RediSep™ silica cartridge) eluting with a gradient of 0 - 8% ethyl acetate in Ao-heptane afforded the desired product as a clear oil (4.13 g, 9.68 mmol, 62%).
XH NMR (400 MHz, DMSO-d6) 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)oxyJ-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
105 mL) and the mixture was heated at 85 °C overnight. The reaction was filtered through celite, eluting with dichloromethane, then concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 80 g RediSep™ silica cartridge) eluting with a gradient of 0 - 8% ethyl acetate in zso-heptane afforded the desired product as a yellow oil (1.92 g, 3.8 mmol, 41%) that was used directly in the subséquent step without characterisation.
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 éthanol (30 mL) was added N-methylthiourca (298 mg, 3.3 mmol, 1 eq) followed by triethylamine (0.92 mL, 6.61 mmol, 2 eq) and the mixture was heated at 80 °C for 4 h. The reaction was allowed to cool to ambient température and the solvent was removed by rotary évaporation. 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 (magnésium sulfate), and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 40 g RediSep™ silica cartridge) eluting with a gradient of 0 - 50% ethyl acetate in zso-heptane afforded the desired product as a brown gum (914 mg, 1.84 mmol, 56%).
LC/MS (C27H36N2O3SiS) 497 [M+H]+; RT 1.46 (LCMS-V-B1)
Ή NMR (400 MHz, DMSO-d6) 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-[(tertbutyldiphenylsilyl)oxy]-2-methylpropyl]-l,3-thiazole-4-carboxylate
To a solution of the product from Step 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 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 température for 2 h. The reaction was partitioned between ethyl acetate and water, and the organic phase was washed with brine, dried (magnésium sulfate), and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 24 g RediSep™ silica cartridge) eluting with a gradient of 0 - 20% ethyl acetate in zso-heptane afforded the desired product as a brown gum (961 mg, 1.61 mmol, 88%).
LC/MS (C32H44N2O5SiS) 597 [M+H]+; RT 1.67 (LCMS-V-B1)
106 φ Ή NMR (400 MHz, DMSO-d6) 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]l,3-thiazole-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 (IM in tetrahydrofuran, 4.83 mL, 4.83 mmol, 3 eq) and the mixture was heated at reflux for 1.5 h. The reaction was allowed to cool to ambient 10 température, then partitioned between ethyl acetate and water, and the organic phase was washed with brine, dried (magnésium sulfate), and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 24 g RediSep™ silica cartridge) eluting with a gradient of 0 - 80% ethyl acetate in Ao-heptane afforded the desired product as a colourless gum (204 mg, 0.57 mmol, 35%).
LC/MS (C16H26N2O5S) 359 [M+H]+; RT 1.07 (LCMS-V-B1) *H NMR (400 MHz, DMSO-d6) 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-[(2S)-3-(2-fluoro-4-iodophenoxy)20 2-methylpropyl]-l,3-thiazole-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-4-iodophenol (211 mg, 0.89 mmol, 2 eq) followed by triphenylphosphine (233 mg, 0.89 mmol, 2 eq) and diisopropylazodicarboxylate (0.17 mL, 0.89 mmol, 2 eq) and the mixture was heated at 120 °C overnight. The reaction was allowed to cool to ambient 25 température and the solvent removed by rotary évaporation. The résultant oil was partitioned between dichloromethane and water, and the organic phase was washed with brine, dried (magnésium sulfate), and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 12 g RediSep™ silica cartridge) eluting with a gradient of 0 - 20% ethyl acetate in Ao-heptane afforded the desired product as a colourless gum (203 mg, 30 0.35 mmol, 79%).
LC/MS (C22H28FIN2O5S) 579 [M+H]+; RT 1.46 (LCMS-V-B1)
107 φ ΧΗ NMR (400 MHz, DMSO-d6) 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), 2.33 - 2.20 (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-l -yn-1 -yl]-2-fluorophenoxy}-2-methylpropyl]-l ,3-thiazole-4carboxylate
To a solution of the product in Step L (226 mg, 0.39 mmol, 1 eq) in tetrahydrofuran (5 mL) was added 3-dimethylamino-l-propyne (0.08 mL, 0.78 mmol, 2 eq), N,N-diisopropylethylamine 10 (0.19 mL, 1.17 mmol, 3 eq) and copper(I) iodide (7.44 mg, 0.04 mmol, 0.1 eq), followed by tetrakis(triphenylphosphine)palladium(0) (45.2 mg, 0.04 mmol, 0.1 eq) and the mixture was heated at reflux for 24 h. The reaction was partitioned between ethyl acetate and water, and the organic phase was washed with brine, dried (magnésium sulfate), and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 24 g RediSep™ 15 silica cartridge) eluting with a gradient of 0 — 5% methanol in dichloromethane afforded the desired product as a brown gum (68 mg, 0.13 mmol, 33%).
LC/MS (C27H36FN3O5S) 534 [M+H]+; RT 1.07 (LCMS-V-B1) XH NMR (400 MHz, DMSO-d6) 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 = 20 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-l-yn-l-yl]-2-fluorophenoxy}-2methylpropyl]-2-(methylamino)-l,3-thiazole-4-carboxylate
To a solution of the product from Step M (68 mg, 0.13 mmol, 1 eq) in dichloromethane (3 25 mL) was added trifluoroacetic acid (0.3 mL, 4.03 mmol, 31.6 eq) and the mixture was stirred at ambient température overnight. The reaction was diluted with dichloromethane, cooled in an ice-bath, and quenched by the addition of saturated aqueous sodium bicarbonate. The organic phase was washed with brine, dried (magnésium sulfate) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 4 g RediSep™ silica 30 cartridge) eluting with a gradient of 0 - 7% methanol in dichloromethane afforded the desired product as a brown gum (40 mg, 0.09 mmol, 72%).
LC/MS (C22H28FN3O3S) 434 [M+H]+; RT 0.82 (LCMS-V-B1)
108 φ Ή NMR (400 MHz, DMSO-d6) 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).
Préparation 3zg: Ethyl 5-(3-{4-[3-(dimethylamino)prop-l-yn-l-yl]-2-fluorophenoxy}2-(oxan-2-yloxy)propyl)-2-(methylamino)-l,3-thiazole-4-carboxylate
Step A: ethyl2-{[(tert-butoxy)carbonyl](methyl)amino}-5-(prop-2-en-l-yl)-l,3-thiazole-4carboxylate
Isopropylmagnesium chloride (2M in tetrahydrofuran; 6 mL, 12 mmol, 1.2 eq) was added dropwise to a solution of ethyl 5-bromo-2-[(tert-butoxycarbonyl)(methyl)amino]-l,3-thiazole4-carboxylate (3.65 g, 9.99 mmol, 1 eq) in dichloromethane (30 mL) at -78 C and the mixture was stirred for 30 min. Zinc chloride solution (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 then for 3 h at ambient température. The solution was cooled in ice-water and copper(I) cyanide di(lithium chloride) complex (IM in tetrahydrofuran; 2 mL, 2 mmol, 0.2 eq) and allyl bromide (2 mL, 23.11 mmol, 2.31 eq) were added slowly. The mixture was stirred for 60 min at 0 °C and then at ambient température for 18 h. Dichloromethane (125 mL) was added and the mixture was successively washed with saturated aqueous ammonium chloride solution (75 mL), water (2 x 75 mL) and brine (75 mL), dried (magnésium sulfate), and concentrated in vacuo. Purification by flash column chromatography (50 g silica) eluting with a gradient of 0 10% ethyl acetate in Ao-heptane afforded the desired product as a colourless gum (1.64 g, 5.02 mmol, 50%).
LC/MS (C15H22N2O4S) 327 [M+H]+; RT 1.43 (LCMS-V-B1) ‘H NMR (400 MHz, DMSO-d6) 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}-l,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 A-bromosuccinimide (774 mg, 4.35 mmol, 1.1 eq), ammonium acetate 30 (30.5 mg, 0.4 mmol, 0.1 eq) and water (4 mL) and the resulting solution was stirred for 45 min at ambient température. Ethyl acetate (150 mL) was added and the mixture was successively washed with water (2 x 75 mL) and brine (75 mL), dried (magnésium sulfate), and concentrated
109 in vacuo. Purification by flash column chromatography (100 g silica) eluting with a gradient of
0-25% ethyl acetate in Ao-heptane afforded the desired product as a colourless gum (960 mg, 2.27 mmol, 57%).
LC/MS (Ci5H23BrN2O5S) 425 [M+H]+; RT 1.29 (LCMS-V-B1)
Ή NMR (400 MHz, DMSO-d6) 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-{[(tertbutoxy)carbonyl](methyl)amino}-l,3-thiazole-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 pyridinium pura-toluenesulphonate (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 température. Dichloromethane (30 mL) was added and the mixture was successively washed with water (2 x 25 mL) and brine (25 mL), dried (magnésium sulfate), and concentrated in vacuo to afford the desired product as a yellow gum (1.52 g, 2.7 mmol, 99%) that was used directly in the next step without further characterisation.
Step D: ethyl 5-[3-(4-bromo-2-fluorophenoxy)-2-(oxan-2-yloxy)propyl]-2-{[(tertbutoxy)carbonyl](methyl)amino}-l,3-thiazole-4-carboxylate
4-Bromo-2-fluorophenol (0.4 mL, 3.65 mmol, 1.35 eq) was added to a suspension of potassium Zeri-butoxide (400 mg, 3.56 mmol, 1.32 eq) in dimethylsulfoxide (5 mL) and the mixture was stirred. A solution of the product from Step C (1.52 g, 2.7 mmol, 1 eq) in dimethylsulfoxide (10 mL) was added and the mixture was stirred for 60 min at ambient température then at 60 °C for 3 h. The reaction was allowed to cool to ambient température and was partitioned between ethyl acetate (150 mL) and water (75 mL), and the organic phase was successively washed with water (2 x 75 mL) and brine (75 mL), dried (magnésium sulfate), and concentrated in vacuo. Purification by flash column chromatography (100 g silica) eluting with a gradient of 0 - 25% ethyl acetate in Ao-heptane afforded the desired product as a pale yellow gum (1.05 g, 1.7 mmol, 63%).
LC/MS (C26H34BrFN2O7S) 617 [M+H]+; RT 1.61 (LCMS-V-B1)
110 φ ‘H NMR (400 MHz, DMSO-d6) 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), 1.49 - 1.38 (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-l5 yn-l-yl]-2-fluorophenoxy}-2-(oxan-2-yloxy)propyl)-l,3-thiazole-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-l-yl)amine (0.25 mL, 2.32 mmol, 1.37 eq), copper(I) iodide (32.4 mg, 0.17 mmol, 0.1 eq) and bis(triphenylphosphine)palladium(II) dichloride (119 mg, 0.17 mmol, 0.1 eq). JV^-Diisopropylethylamine (0.9 mL, 5.1 mmol, 3 eq) was added and 10 the mixture was heated at 75 °C for 24 h. The reaction was allowed to cool to ambient température and was partitioned between ethyl acetate (150 mL) and water (75 mL), and the organic phase was successively washed with water (75 mL) and brine (75 mL), dried (magnésium sulfate), and concentrated in vacuo. Purification by flash column chromatography (50 g silica) eluting with a gradient of 0 -100% ethyl acetate in iso-heptane afforded the desired 15 product as a brown gum (395 mg, 0.64 mmol, 38%).
LC/MS (C3iH42FN3O7S) 620 [M+H]+; RT 1.31 (LCMS-V-B1)
Step F: ethyl 5-(3-{4-[3-(dimethylamino)prop-l-yn-l-yl]-2-fluorophenoxy}-2-(oxan-2yloxy)propyl)-2-(methylamino)-l,3-thiazole-4-carboxylate
To a solution of the product in Step E (390 mg, 0.63 mmol, 1 eq) in 1,1,1,3,3,320 hexafluoropropan-2-ol (7 mL) was heated in a sealed tube at 100 °C for 60 min then allowed to cool to ambient température and concentrated in vacuo. Purification by flash column chromatography (20 g silica) eluting with a gradient of 0 - 2.5% methanol in dichloromethane afforded the desired product as a brown gum (170 mg, 0.33 mmol, 52%).
LC/MS (C26H34FN3O5S) 520 [M+H]+; RT 1.05 (LCMS-V-B1)
Préparation 3zh: Ethyl 5-(2-methoxy-3-{[tris(propan-2-yl)silyl]oxy}propyl)-2(methylamino)-l,3-thiazole-4-carboxyIate
Step A: ethyl2-{[(tert-butoxy)carbonyl](methyl)amino}-5-(2,3-dihydroxypropyl)-l,3thiazole-4-carboxylate
To a solution of the product from Préparation 3zg, Step A (8.39 g, 25.7 mmol, 1 eq) in 9:1 acetone / water (250 mL) was added 4-methylmorpholine-A-oxide (4.75 g, 40.6 mmol, 1.58
111 eq), followed by osmium tetroxide (3.5 mL, 0.28 mmol, 0.01 eq) and the resulting mixture was stirred for 18 h at ambient température. Water (300 mL) was added and the mixture was extracted with ethyl acetate (4 x 200 mL), and the combined organics were washed with brine (150 mL), dried (magnésium sulfate), and concentrated in vacuo. Purification by flash column chromatography (100 g silica) eluting with a gradient of 0 — 5% methanol in dichloromethane afforded the desired product as a brown gum (8.66 g, 24 mmol, 94%).
LC/MS (Ci5H24N2O6S) 361 [M+H]+; RT 1.07 (LCMS-V-B1) ]H NMR (400 MHz, DMSO-d6) 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 ofthe product from Step A (8.65 g, 24 mmol, 1 eq) in dichloromethane (100 mL), cooled in ice-water, was added imidazole (3.5 g, 51.4 mmol, 2.14 eq), followed by triisopropylsilyl chloride (6 mL, 28 mmol, 1.17 eq) dropwise and the mixture was stirred for 60 min at 0 °C then for 18 h at ambient température. The reaction was concentrated in vacuo and purification by flash column chromatography (100 g silica) eluting with a gradient of 0 25% ethyl acetate in /so-heptanc afforded the desired product as a colourless oil (12.2 g, 23.5 mmol, 98%).
LC/MS (C24H44N2O6SiS) 517 [M+H]+; RT 1.69 (LCMS-V-B1)
Tl NMR (400 MHz, DMSO-d6) δ 5.07 (d, J = 5.2 Hz, 1H), 4.24 (q, J = 7.1 Hz, 2H), 3.75 3.61 (m, 2H), 3.55 - 3.48 (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)-l,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 AWW’W’-tetramethylnaphthalene1,8-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 then for 24 h at ambient température. The mixture was diluted with dichloromethane (75 mL) and washed successively with IM aqueous copper(II) sulphate (2 x 75 mL), water (75 mL) and brine (75 mL), dried (magnésium sulfate), and concentrated in
112 vacuo. Purification by flash column chromatography (50 g silica) eluting with a gradient of 0 5% ethyl acetate in zso-heptane afforded the desired product as a colourless gum (2.54 g, 4.79 mmol, 95%).
LC/MS (C25H46N2O6SiS) 531 [M+H]+; RT 1.80 (LCMS-V-B1)
Ή NMR (400 MHz, DMSO-d6) δ 4.37 (q, J = 7.1 Hz, 2H), 3.75 (dd, J = 5.0, 2.6 Hz, 2H), 3.55 (s, 3H), 3.52 - 3.44 (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)-l,3thiazole-4-carboxylate
A solution of the product from Step C (2.54 g, 4.79 mmol, 1 eq) in 1,1,1,3,3,3hexafluoropropan-2-ol (20 mL) was heated in a sealed tube at 100 °C for 5 h. The solution was allowed to cool to ambient température, concentrated in vacuo and dried under vacuum to afford the desired product as a white solid (1.75 g, 4.06 mmol, 85%).
Ή NMR (400 MHz, DMSO-d6) 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).
Préparation 4a: \-(6-Chl()r()-4-methvl-pyridazin-3-yl)-3-(2trimethylsilylethoxymethyl)-l,3-benzothiazol-2-imine
Step A: N-(6-chloro-4-methyl-pyridazin-3-yl)-l,3-benzothiazol-2-amine
A 2 L oven-dried, one-necked, round-bottom flask was equipped with a PTFE-coated magnetic stirring bar and fitted with a reflux condenser. It was charged with 34.0 g of 6-chloro-4-methylpyridazin-3-amine (237 mmol, 1 eq.), 34 mL of 2-chloro-l,3-benzothiazole (44.2 g, 260 mmol, 1.1 eq.), 124 mL of DIPEA (91.8 g, 710 mmol, 3 eq.) and 137 g of Cs2CO3 (710 mmol, 3 eq.), then 1 L of DMF were added and the system was flushed with argon. After 5 minutes stirring under inert atmosphère 2.01 g of Pd3(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 warmed up to 75°C and stirred at that température for 4 hours to reach complété conversion. Reaction mixture was left to cool down to rt, then poured into 3 L of water while it was intensively stirred. After 30 min the precipitated product was removed by filtration, and then it was washed with water for 2 times (2x2 L). The product was dried overnight on high vacuum. The dried crude product was stirred
113 in 1 L of heptane : Et2O (3:2) for 30 min then filtered off to give 64.5 g (98%) of the desired product as green powder.
Ή NMR (500 MHz, DMSO-d6) 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); 13C NMR (125 MHz, DMSO-d6) δ ppm 130.3, 129.5, 126.6, 122.8, 122.3, 17.2; HRMS-ESI (m/z): [M+H]+ calcd for C12H10CIN4S: 277.0309, found 277.0305.
Step B : N-(6-chloro-4-methvl-pyridazin-3-yl) -3-(2-trimethylsilylethoxymethyl) -1,3benzothiazol-2-imin e
A 2 L oven-dried, one-necked, round-bottomed flask equipped with a PTFE-coated magnetic stirring 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.), 14.43 g of N,N-dimethylpyridin-4-amine (11.81 mmol, 0.05 eq.) in 1 L of dry DCM were cooled down to 0 °C under N2. And during intensive mechanical stirring 46.00 mL of 2-(chloromethoxy)ethyl-trimethyl-silane (43.32 g, 259 mmol, 1.1 eq.) was added to the mixture dropwise over 5 min period of time. It was stirred at 0°C for 30 min when the reaction reached complété conversion. 24.5 mL of water was added to the reaction mixture then Celite was added to the reaction mixture and the volatiles were removed under reduced pressure. It was purified via flash column chromatography using heptane and EtOAc as eluents to obtain 46.62 g (48%) of the desired product.
XH NMR (500 MHz, DMSO-d6) 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); 13C NMR (125 MHz, DMSO-d6) 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; HRMS-ESI (m/z): [M+H]+ calcd for Ci8H24ClN4OSSi: 407.1123, found 407.1120.
Préparation 4b: (2Z)-/V-(6-ChIoropyridazin-3-yl)-3-{[2(trimethylsilyl)ethoxy]methyl}-2,3-dihydro-l,3-benzothiazol-2-imine
N,A-Diisopropylethylamine (3 mL, 16.6 mmol, 3 eq) was added to a suspension of the product from Préparation 9b (1.45 g, 5.52 mmol, 1 eq) in dichloromethane (75 mL) at 0 °C under a nitrogen atmosphère. [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 température for 3 h. Dichloromethane (75 mL) was added and the mixture washed with water (2 x 75 mL) and brine (75 mL). The solution was dried (magnésium sulfate)
114 and concentrated in vacuo. Trituration with methanol (20 mL) gave a solid that was filtered, washed with methanol (2 x 10 mL) and dried under vacuum to afford the desired product as a pale brown solid (1.85 g, 4.71 mmol, 85%).
LC/MS (Ci7H2iClN4OSiS) 393 [M+H]+; RT 1.56 (LCMS-V-B1).
Préparation 5a: Methyl 5-[3-[4-[3-[terCbutoxycarbonyl(methyl)amino]prop-l-ynyl]2-fluoro-phenoxy]propyI]-2-[[4-[tert-butyl(diphenyl)siIyl]oxy-5-(ptolylsulfonyloxy)pentyl]-[5-methyl-6-[(Z)-[3-(2-trimethylsilylethoxymethyl)-l,3benzothiazol-2-ylidene]amino]pyridazin-3-yl]amino]thiazole-4-carboxylate
Step A: methyl 5-[3-[4-[3-[tert-butoxycarbonyl(methyl)amino]prop-l-ynyl]-2-fluorophenoxy]propyl]-2-[[5-[tert-butyl(dimethyl)silyl]oxy-4-[tert-butyl(diphenylsilyl]oxy-pentyl][5-methyl-6-[(Z)-[3-(2-trimethylsilylethoxymethyl)-l,3-benzothiazol-2ylidene]amino]pyridazin-3-yl]amino]thiazole-4-carboxylate
Using Buchwald General Procedure II starting from 12 g of Préparation 3a (13 mmol) and 6.30 g of Préparation 4a (15.6 mmol) as the appropriate halide, 14 g (83%) of the desired product was obtained.
]H NMR (500 MHz, DMSO-d6) 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), 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); 13C NMR (125 MHz, DMSO-d6)ôppm 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; HRMS-ESI (m/z): [M+H]+ calcd for CesHçsF^OsSzSis: 1302.5813, found 1302.5819.
Step B: methyl 5-[3-[4-[3-[tert-butoxycarbonyl(methyl)amino]prop-l-ynyl]-2-fluorophenoxy]propyl]-2-[[4-[tert-butyl(diphenyl)silyl]oxy-5-hydroxy-pentyl]-[5-methyl-6-[(Z)-[3(2-trimethylsilylethoxymethyl)-l,3-benzothiazol-2-ylidene]amino]pyridazin-3yl]amino]thiazole-4-carboxylate
A 100 mL oven-dried, one-necked, round-bottom flask was equipped with a PTFE-coated magnetic stirring bar and fitted with a reflux condenser. It was charged with 1.40 g of the product from Step A (1.1 mmol, 1 eq.) and 12 mg of camphor sulfonic acid (0.054 mmol, 0.05 eq.), 5 mL of DCM and 1 mL of MeOH. The resulting mixture was stirred overnight at rt to
115 reach complété conversion. Reaction mixture was concentrated directly to Celite then purified by flash column chromatography using heptane and EtOAc as eluents to give 700 mg (55%) of the desired product as yellow solid.
Ή NMR (500 MHz, DMSO-dô) 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); 13C NMR (125 MHz, DMSO-dô) 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, 27.2, 23.1, 22.5,17.9,17.8, -1.0; HRMS-ESI (m/z): [M+H]+ calcd for Cô2H79FN7O8S2SÎ2: 1188.4949, found 1188.4938.
Step C: methyl 5-[3-[4-[3-[tert-butoxycarbonyl(methyl)amino]prop-l-ynyl]-2-fluorophenoxy]propyl]-2-[[4-[tert-butyl(diphenyl)silyl]oxy-5-(p-tolylsulfonyloxy)pentyl]-[5methyl-6-[(Z)-[3-(2-trimethylsilylethoxymethyl)-l,3-benzothiazol-2ylidene]amino]pyridazin-3-yl]amino]thiazole-4-carboxylate
A 100 mL oven-dried, one-necked, round-bottom flask was equipped with a PTFE-coated magnetic stirring bar was charged with 700 mg of the product from Step B (0.58 mmol, 1 eq.) and 907 mg of N,N-dimethyl-l-(p-tolylsulfonyl)pyridin-l-ium-4-amine chloride (2.9 mmol, 5 eq., Tetrahedron Lett. 2016, 57, 4620) were dissolved in 35 mL of DCM and stirred overnight at rt. Reaction reached complété conversion. Reaction mixture directly was concentrated onto Celite, and then purified by flash column chromatography using heptane and EtOAc as eluents to give 450 mg (56%) of the desired product.
Ή NMR (500 MHz, DMSO-dô) 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); 13C NMR (125 MHz, DMSO-dô) 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; HRMS-ESI (m/z): [M+H]+ calcd for C69H85FN7OioS3Si2: 1342.5037, found 1342.5039.
Préparation 5b: Methyl 2-[[4-[terAbutyI(diphenyl)silyl]oxy-5-(ptolylsulfonyloxy)pentyl]-[5-methyl-6-[(Z)-[3-(2-trimethylsilylethoxymethyl)-l,320487
116 benzothiazol-2-yIidene]amino]pyridazin-3-yl]amino]-5-[3-[4-[3-(dimethylamino)prop-lynyl]-2-fluoro-phenoxy]propyl]thiazole-4-carboxylate
Step A: methyl 2-[[5-[tert-butyl(dimethyl)silyl]oxy-4-[tert-butyl(diphenyl)silyl]oxy-pentyl][5-methyl-6-[(Z)-[3-(2-trimethylsilylethoxymethyl)-l,3-benzothiazol-25 ylidene]amino]pyridazin-3-yl]amino]-5-[3-[4-[3-(dimethylamino)prop-l-ynyl]-2-fluorophenoxy]propyl]thiazole-4-carboxylate
Using Buchwald General Procedure II starting from 6.0 g of Préparation 3b (7.1 mmol, 1 eq.) and 3.46 g of Préparation 4a (8.51 mmol, 1.2 eq.) as the appropriate halide, 7.0 g (81%) of the desired product was obtained.
Ή NMR (500 MHz, DMSO-d6) 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); 13C NMR (125 MHz, DMSO-d6) δ ppm 135.9-111.3,127.2-112.0,128.9, 15 119.2, 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; HRMS-ESI (m/z): [M+H]+ calcd for CôÆstFNtOôSzSîj: 1216.5446, found 1216.5425.
Step B: methyl 2-[[4-[tert-butyl(diphenyl)silyl]oxy-5-hydroxy-pentyl]-[5-methyl-6-[(Z)-[3-(2trimethylsilylethoxymethyl)-l,3-benzothiazol-2-ylidene]amino]pyridazin-3-yl]amino]-5-[320 [4-[3-(dimethylamino)prop-l-ynyl]-2-fluoro-phenoxy]propyl]thiazole-4-carboxylate
A 100 mL oven-dried, one-necked, round-bottom flask was equipped with a PTFE-coated magnetic stirring bar and fitted with a reflux condenser. It was charged with 7.0 g of the product from Step A (5.75 mmol, 1 eq.) and 1.60 g of camphor sulfonic acid (6.90 mmol, 1.2 eq.) dissolved in 25 mL of DCM and 5 mL of MeOH. The resulting mixture was stirred overnight 25 at rt to reach complété conversion. Reaction mixture was concentrated directly to Celite, and purified by flash column chromatography using DCM and MeOH (1.2% NH3) as eluents to give 3.0 g (47%) of the desired product as yellow solid.
Step C: methyl 2-[[4-[tert-butyl(diphenyl)silyl]oxy-5-(p-tolylsulfonyloxy)pentyl]-[5-methyl6-[(Z)-[3-(2-trimethylsilylethoxymethyl)-l,3-benzothiazol-2-ylidene]amino]pyridazin-330 yl]amino]-5-[3-[4-[3-(dimethylamino)prop-l-ynyl]-2-fluoro-phenoxy]propyl]thiazole-4carboxylate
A 100 mL oven-dried, one-necked, round-bottom flask was equipped with a PTFE-coated
117 magnetic stirring bar was charged with 3.80 g of the product from Step B (3.40 mmol, 1 eq.),
5.40 g oî N,N-dimethyl-l-(p-tolylsulfonyl)pyridin-l-iiim-4-amine chloride (17.0 mmol, 5 eq., Tetrahedron Lett. 2016, 57, 4620) and 20 mL of DCM then stirred overnight at rt. Reaction reached complété conversion. Reaction mixture was concentrated directly to Celite then purified by flash column chromatography using DCM and MeOH (1.2% NH3) as eluents to give 3.58 g (83%) of the desired product.
Ή NMR (500 MHz, DMSO-dô) 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); 13C NMR (125 MHz, DMSO-d6) 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; HRMS-ESI (m/z): [M+H]+ calcd for C65H79FN7O8S3 Si2: 1256.4669, found 1256.4677.
Préparation 5c: Methyl 5-[3-[4-[3-[ierAbutoxycarbonyl(methyl)amino]prop-l-ynyl]2-fluoro-phenoxy]propyl]-2-[4-hydroxybutyI-[5-methyl-6-[(Z)-[3-(2trimethylsilylethoxymethyl)-l,3-benzothiazol-2-yIidene]amino]pyridazin-3yl]amino]thiazole-4-carboxylate
Step A: methyl 5-[3-[4-[3-[tert-butoxycarbonyl(methyl)amino]prop-l-ynyl]-2-fluorophenoxy]propyl]-2-[4-[tert-butyl(dimethyl)silyl]oxybutyl-[5-methyl-6-[(Z)-[3-(2trimethylsilylethoxymethyl)-l,3-benzothiazol-2-ylidene]amino]pyridazin-3yl]amino]thiazole-4-carboxylate
Using Buchwald General Procedure II starting from 250 mg of Préparation 3c (0.38 mmol, 1 eq.) and 250 mg of Préparation 4a (0.38 mmol, 1 eq.) as the appropriate halide, 295 mg (75%) of the desired product was obtained.
'H NMR (500 MHz, DMSO-dô) 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); 13C NMR (125 MHz, DMSOdô) 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; HRMS-ESI (m/z): [M+H]+
118 calcd for C51H73FN7O7S2S12: 1034.4530, found 1034.4517.
Step B: methyl 5-[3-[4-[3-[tert-butoxycarbonyl(methyl)amino]prop-l-ynyl]-2-fluorophenoxy]propyl]-2-[4-hydroxybutyl-[5-methyl-6-[(Z)-[3-(2-trimethylsilylethoxymethyl)-l,3benzothiazol-2-ylidene]amino]pyridazin-3-yl]amino]thiazole-4-carboxylate
Using Deprotection of tert-butyl-dimethyl-silyl protecting group General Procedure starting from 650 mg of the product from Step A (0.63 mmol, 1 eq.) and 36 mg of [(lS,4R)-7,7dimethyl-2-oxo-norbornan-l-yl]methanesidfonic (0.16 mmol, 0.25 eq.), 178 mg (31%) of the desired product was obtained.
Ή NMR (500 MHz, DMSO-d6) δ ppm 7.84 (d, 1H), 7.68 (s, 1H), 7.46 (d, 1H), 7.43 (td, 1H), 10 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); 13C NMR (125 MHz, DMSO-dô) Sppm 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, 24.1, 23.2,17.9,17.8, -1.0; HRMS-ESI 15 (m/z): [M+H]+ calcd for C45H59FN7O7S2S1: 920.3665, found 920.3650.
Préparation 5d: Methyl 5-[3-[4-[3-(dimethylamino)prop-l-ynyl]-2-fluorophenoxy]propyl]-2-[4-hydroxybutyl-[5-methyl-6-[(Z)-[3-(2-trimethylsiIylethoxymethyl)l,3-benzothiazoI-2-ylidene]amino]pyridazin-3-yl]amino]thiazole-4-carboxylate
Step A: methyl 2-[4-[tert-butyl(dimethyl)silyl]oxybutyl-[5-methyl-6-[(Z)-[3-(220 trimethylsilylethoxymethyl)-l,3-benzothiazol-2-ylidene]amino]pyridazin-3-yl]amino]-5-[3[4-[3-(dimethylamino)prop-l-ynyl]-2-fluoro-phenoxy]propyl]thiazole-4-carboxylate
Using Buchwald General Procedure II starting from 1800 mg of Préparation 3d (3.11 mmol, 1 eq.) and 1267 mg of Préparation 4a (3.11 mmol, 1 eq.) as the appropriate halide, 2035 mg (69%) of the desired product was obtained.
Ή NMR (500 MHz, DMSO-d6) δ ppm 7.82 (dm, 1H), 7.61 (s, 1H), 7.44 (dm, 1H), 7.42 (m, 1H), 7.29 (dd, 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); 13C NMR (125 MHz, DMSO-d6) δ ppm 128.9, 127.1, 123.4, 123.2, 119.2, 117.5,
119 φ 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, 23.7, 23.1, 17.9, 17.8, -1.0, -4.9; HRMS-ESI (m/z): [M+H]+ calcd for C47H67FN7O5S2Si2: 948.4162, found 948.4161.
Step B: methyl 5-[3-[4-[3-(dimethylamino)prop-l-ynyl]-2-fluoro-phenoxy]propyl]-2-[45 hydroxybutyl-[5-methyl-6-[(Z)-[3-(2-trimethylsilylethoxymethyl)-l,3-benzothiazol-2ylidene]amino]pyridazin-3-yl]amino]thiazole-4-carboxylate
Using Deprotection of tert-butyl-dimethyl-silyl protecting group General Procedure starting from 7.23 g of the product from Step A (7.63 mmol, 1 eq.) and 2.22 g of [(lS,4R)-7,7dimethyl-2-oxo-norbornan-l-yl]methanesulfonic (9.54 mmol, 1.25 eq.), 5.23 g (82%) of the 10 desired product was obtained.
’H NMR (500 MHz, DMSO-ifo) 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); 13C NMR 15 (125 MHz, DMSO-Jô) 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; HRMS-ESI (m/z): [M+H]+ calcd for C4iH53FN7O5S2Si: 834.3297, found 834.3296.
Préparation 5e: Methyl 5-[3-[4-[3-[terributoxycarbonyl(methyI)amino]prop-l-ynyl]2-fluoro-phenoxy]propyl]-2-[5-hydroxypentyl-[5-methyl-6-[(Z)-[3-(220 trimethylsilylethoxymethyl)-l,3-benzothiazol-2-ylidene]amino]pyridazin-3yl]amino]thiazole-4-carboxylate
Step A: methyl 5-[3-[4-[3-[tert-butoxycarbonyl(methyl)amino]prop-l-ynyl]-2-fluorophenoxy]propyl]-2-[5-[tert-butyl(dimethyl)silyl]oxypentyl-[5-methyl-6-[(Z) -[3-(2trimethylsilylethoxymethyl)-l,3-benzothiazol-2-ylidene]amino]pyridazin-325 yl]amino]thiazole-4-carboxylate
Using Buchwald General Procedure II starting from 350 mg of Préparation 3m (0.52 mmol, 1 eq.) and 210 mg of Préparation 4a (0.52 mmol, 1 eq.) as the appropriate halide, 481 mg (88%) of the desired product was obtained.
*H NMR (500 MHz, DMSO-d6) δ ppm 7.81 (dd, 1H), 7.62 (s, 1H), 7.45 (dd, 1H), 7.42 (m, 1H), 30 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),
120 φ 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); 13C NMR (125 MHz, DMSO-d6) 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, 62.7, 51.8, 47.0, 38.6, 33.8, 5 32.4, 31.0, 28.5, 27.1, 26.2, 23.1, 23.0, 17.9, 17.8, -1.0, -5.0; HRMS-ESI (m/z): [M+H]+ calcd for C52H75FN7O7S2Si2: 1048.4686, found 1048.4692.
Step B: methyl 5-[3-[4-[3-[tert-butoxycarbonyl(methyl)amino]prop-l-ynyl]-2-fluorophenoxy]propyl]-2-[5-hydroxypentyl-[5-methyl-6-[ (Z) -[3-(2-trimethylsilylethoxymethyl) -1,3benzothiazol-2-ylidene]amino]pyridazin-3-yl]amino]thiazole-4-carboxylate
Using Deprotection of tert-butyl-dimethyl-silyl protecting group General Procedure starting from 2.52 g of the product from Step A (2.40 mmol, 1 eq.) and 0.70 g of [(lS,4R)-7,7dimethyl-2-oxo-norbornan-l-yl]methanesulfonic (3.00 mmol, 1.25 eq.), 1.19 g (53%) of the desired product was obtained.
]H NMR (500 MHz, DMSO-d6) δ ppm 7.84 (d, 1H), 7.66 (s, 1H), 7.46 (d, 1H), 7.43 (t, 1H), 15 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); 13C NMR (125 MHz, DMSO-îA) 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-ESI (m/z): [M+Na]+ calcd for C46H6oFN7Na07S2 Si: 956.3641, found 956.3646.
Préparation 5f: Methyl 5-[3-[4-[3-(dimethylamino)prop-l-ynyl]-2-fluorophenoxy]propyl]-2-[5-hydroxypentyl-[5-methyl-6-[(Z)-[3-(2-trimethylsilylethoxymethyl)l,3-benzothiazol-2-ylidene]amino]pyridazin-3-yl]amino]thiazole-4-carboxylate
Step A: methyl 2-[5-[tert-butyl(dimethyl)silyl]oxypentyl-[5-methyl-6-[(Z)-[3-(225 trimethylsilylethoxymethyl)-l,3-benzothiazol-2-ylidene]amino]pyridazin-3-yl]amino]-5-[3[4-[3-(dimethylamino)prop-l-ynyl]-2-fluoro-phenoxy]propyl]thiazole-4-carboxylate
Using Buchwald General Procedure II starting from 3.68 g of Préparation 3t (6.22 mmol, 1 eq.) and 3.29 g of Préparation 4a (8.08 mmol, 1.3 eq.) as the appropriate halide, 3.92 g (65%) of the desired product was obtained.
*H NMR (500 MHz, DMSO-d6) δ ppm 7.83 (dm, 1H), 7.66 (s, 1H), 7.46 (dm, 1H), 7.43 (m,
121 φ 1Η), 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); HRMS-ESI (m/z): [M+H]+ calcd for C48H69FN7O5S2Si2:962.4319, found 5 962.4301.
Step B: methyl 5-[3-[4-[3-(dimethylamino)prop-l-ynyl]-2-fluoro-phenoxy]propyl]-2-[5hydroxypentyl-[5-methyl-6-[(Z) -[3-(2-trimethylsilylethoxymethyl) -1,3-benzothiazol-2 ylidene]amino]pyridazin-3-yl]amino]thiazole-4-carboxylate
Using Deprotection of tert-butyl-dimethyl-silyl protecting group General Procedure 10 starting from 3.91 g of the product from Step A (4.06 mmol, 1 eq.) and 1.18 g of [(lS,4R)-7,7dimethyl-2-oxo-norbornan-l-yl]methanesulfonic (5.08 mmol, 1.25 eq.), 2.61 g (76%) of the desired product was obtained.
Ή NMR (500 MHz, DMSO-î/6) 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 15 (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); 13C NMR (125 MHz, DMSO-t/6) 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; HRMS-ESI (m/z): [M+2H]2+ calcd for C42H56FN7O5S2Si: 424.6764, found 20 424.6755.
Préparation 5g: Ethyl 5-(3-iodopropyl)-2-[methyl-[5-methyl-6-[(Z)-[3-(2trimethylsilylethoxymethyl)-l,3-benzothiazol-2-ylidene]amino]pyridazin-3y 1] amino] thiazole-4-carboxylate
Step A: ethyl5-(3-chloropropyl)-2-[methyl-[5-methyl-6-[(Z)-[3-(225 trimethylsilylethoxymethyl)-l,3-benzothiazol-2-ylidene]amino]pyridazin-3yl]amino]thiazole-4-carboxylate
Using Buchwald General Procedure II starting from 3.15 g of Préparation 3e (12 mmol, 1.2 eq.) and 4.07 g of Préparation 4a (10 mmol, 1 eq.) as the appropriate halide, 2.6 g (41%) of the desired product was obtained.
Ή NMR (500 MHz, DMSO-d6) δ ppm 7.84 (d, 1H), 7.65 (s, 1H), 7.45 (d, 1H), 7.43 (tm, 1H),
122 φ 7.25 (tm, 1Η), 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); 13C NMR (125 MHz, DMSO-dô) 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;
HRMS-ESI (m/z): [M+H]+ calcd for C28H38ClN6O3S2Si: 633.1899, found 633.1891.
Step B: ethyl 5-(3-iodopropyl)-2-[methyl-[5-methyl-6-[(Z)-[3-(2-trimethylsilylethoxymethyl)fS-benzothiazol^-ylideneJanûnolpyridazin-S-ylJarninoJthiazole-d-carboxylate
A 100 mL one-necked, round-bottomed flask was equipped with a PZFE-coated magnetic stirring bar and fitted with a reflux condenser. It was charged with 2.6 g of the product from 10 Step A (4.10 mmol, 1 eq.), 1.23 g of Nal (8.2 mmol, 2 eq.) and 20 mL of dry acetone. The reaction mixture was warmed up to 60°C and stirred at that température for 3 days, when the reaction reached complété conversion. The reaction mixture was diluted with the addition of water then the precipitated product was collected by filtration, washed with water, and then dried on high vacuum to obtain 2.5 g (84%) of the desired product.
Ή NMR (500 MHz, DMSO-d6) 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); 13C NMR (125 MHz, DMSO-dô) 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; HRMS20 ESI (m/z): [M+H]+ calcd for C28H38I NôO3S2Sî: 725.1255, found 725.1248.
Préparation 5h: Ethyl 5-[3-[4-[3-[tert-butoxycarbonyl-[2(dimethylamino)ethyl]amino]prop-l-ynyl]-2-fluoro-phenoxy]propyl]-2-[methyl-[5methyl-6-[(Z)-[3-(2-trimethylsilylethoxymethyl)-l,3-benzothiazol-2ylidene]amino]pyridazin-3-yl]amino]thiazole-4-carboxylate
Step A: tert-butylN-[2-(dimethylamino)ethyl]-N-prop-2-ynyl-carbamate
365 mg oîN',N'-dimethyl-N-prop-2-ynyl-ethane-l,2-diamine (1.83 mmol, 1 eq.) and 22 mg of N,N-dimethylpyridin-4-amine (0.18 mmol, 0.1 eq.) were mixed in dichloromethane (2.2 mL/mmol) then 600 mg of tert-butoxycarbonyl tert-butyl carbonate (2.75 mmol, 1.5 eq.) was added in one portion at rt and the mixture was and stirred at ambient température for 3 h. After 30 the reaction time ca. 10 mL of DCM and ca. 10 mL of cc. NaHCOs were added, separated and the organic layer was dried over MgSO4, filtered and concentrated. Yellowish oil was obtained
123 as crude product: 332 mg (77%) of the desired product.
Ή NMR (400 MHz, DMSO-de) 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); 13C NMR (100 MHz, DMSO-d6) δ ppm 149.3, 106.7, 79.2, 57.0, 45.3, 43.7, 38.6, 28.0.
Step B: (2-fluoro-4-iodo-phenoxy)-triisopropyl-silane
A 100 mL oven-dried, one-necked, round-bottomed flask was equipped with a PTFE-coated magnetic stirring bar. It was charged with 2.38 g of 2-fluoro-4-iodo-phenol (10.00 mmol), 1.98 g of K2CO3 (20.00 mmol, 2 eq.) and acetonitrile (4 mL/mmol). To the resulting mixture 2.31 g of chloro(triisopropyl)silane (12.00 mmol, 1.2 eq.) was added dropwise near intensive stirring at rt. The resulting mixture was stirred at rt for 30 min, while the reaction reached complété conversion. The reaction mixture was filtered through a pad of Celite then concentrated onto Celite and purified via flash column chromatography using heptane and EtOAc as eluents to give 3.70 g (94%) of the desired product (94%) as a colorless oil.
Ή NMR (500 MHz, DMSO-d6) δ ppm 7.60 (dd, 1H), 7.40 (dm, 1H), 6.82 (dd, 1H), 1.24 (sp, 3H), 1.04 (d, 18H); 13C NMR (125 MHz, DMSO-de) δ ppm 153.9, 143.8, 134.2, 125.5, 124.1, 83.6, 18.0, 12.5; HRMS-EI (m/z): [M]+ calcd for C15H24FIOS1: 394.0625, found 394.0616.
Step C: tert-butyl N-[2-(dimethylamino)ethyl]-N-[3-(3-fluoro-4-triisopropylsilyloxyph enyl) prop-2-ynyl] carbamate
Using Sonogashira General 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 to give 90 mg (75%) of the desired product as yellow oil.
Ή NMR (400 MHz, DMSO-d6) 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); 13C NMR (100 MHz, DMSO-ώ) 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-ESI (m/z): [M+H]+ calcd for C27H46FN2O3Si: 493.3256, found 493.3264.
Step D: ethyl 5-[3-[4-[3-[tert-butoxycarbonyl-[2-(dimethylamino)ethyl]amino]prop-l-ynyl]2-fluoro-phenoxy]propyl]-2-[methyl-[5-methyl-6-[(Z)-[3-(2-trimethylsilylethoxymethyl)-l,3benzothiazol-2-ylidene]amino]pyridazin-3-yl]amino]thiazole-4-carboxylate
Using Alkylation of SilyLProtected Phénols General Procedure starting from 147 mg of Préparation 5g, Step B (0.20 mmol, 1 eq.) and 100 mg of the product from Step C (0.20 mmol, 1 eq.) to give 164 mg (87%) of the desired product as brown solid.
124 φ Ή NMR (500 MHz, DMSO-î/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); 13C NMR 5 (125 MHz, DMSO-ά) 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; HRMS-ESI (m/z): [M+H]+ calcd for C46H62FN8O6S2Si: 933.3982, found 933.3977.
Préparation 5i: Methyl 5-[3-[4-[3-(terAbutoxycarbonylamino)prop-l-ynyl]-210 fluoro-phenoxy]propyl]-2-[4-[terAbutyl(dimethyI)silyl]oxybutyl-[5-methyl-6-[(Z)-[3-(2trimethyIsilylethoxymethyl)-l,3-benzothiazoI-2-ylidene]amino]pyridazin-3yl]amino]thiazole-4-carboxylate
Using Buchwald General Procedure II starting from 780 mg of Préparation 3g (1.20 mmol, 1 eq.) and 489 mg of Préparation 4a (1.20 mmol, 1 eq.) as the appropriate halide, 570 mg 15 (47%) of the desired product was obtained.
>H NMR (500 MHz, DMSO-d6) 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), 20 0.12 (s, 9H); 13C NMR (125 MHz, DMSO-d6) 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; HRMS-ESI (m/z): [M+H]+ calcd for CsoHviFNïChSzSis: 1020.4373, found 1020.4373.
Préparation 5i: Ethyl 5-(3-{2-fluoro-4-[3-(methyiamino)prop-l-yn-l25 yl]phenoxy}propyl)-2-[methyl(5-inethyl-6-{[(2Z)-3-{[2-(trimethyIsilyl)ethoxy]methyl}2,3-dihydro-l,3-benzothiazol-2-ylidene]amino}pyridazin-3-yl)amino]-l,3-thiazole-4carboxylate
Step A: ethyl5-{3-[4-(3-{[(tert-butoxy)carbonyl](methyl)amino}prop-l-yn-l-yl)-2fluorophenoxy]propyl}-2-[methyl(5-methyl-6-{[(2Z)-3-{[2-(trimethylsilyl)ethoxy]methyl}20487
125
2,3-dihydro-l,3-benzothiazol-2-ylidene]amino}pyridazin-3-yl)amino]-l,3-thiazole-4carboxylate
To the product from Préparation 5g (1.75 g, 2.41 mmol, 1 eq) in dimethylformamide (50 mL) was added the product from Préparation 6a (877 mg, 3.14 mmol, 1.3 eq) in dimethylformamide (10 mL) and césium carbonate (2.36 g, 7.24 mmol, 3 eq) and the mixture was heated at 80 °C for 16 h. The reaction was concentrated in vacuo then partitioned between ethyl acetate and brine, and the organic phase was dried (magnésium sulfate) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 40 g RediSep™ silica cartridge) eluting with a gradient of 0 — 50% ethyl acetate in iso-heptane afforded the desired product as a yellow oil (1.75 g, 2 mmol, 83%).
LC/MS (C43H54FN7O6SiS2) 876 [M+H]+; RT 1.46 (LCMS-V-B2) ‘H NMR (400 MHz, DMSO-d6) 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-l-yn-1 -yl]phenoxy}propyl)-2[methyl(5-methyl-6-{[(2Z)-3-{[2-(trimethylsilyl)ethoxy]methyl}-2,3-dihydro-l,3benzothiazol-2-ylidene]amino}pyridazin-3-yl)amino]-l,3-thiazole-4-carboxylate
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 température for 5 h. The reaction was diluted with dichloromethane, cooled to 0 °C and basified by the addition of 2N aqueous sodium hydroxide. The organic phase was dried (magnésium sulfate) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 40 g RediSep™ silica cartridge) eluting with a gradient of 0 - 10% methanol in dichloromethane afforded the desired product as a yellow gum (329 mg, 0.42 mmol, 25%).
LC/MS (C38H46FN7O4SiS2) 776 [M+H]+; RT 2.58 (LCMS-V-C)
Ή NMR (400 MHz, DMSO-d6) 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).
126
Préparation 5k: Ethyl 2-[methyl(5-methyl-6-{[(2Z)-3-{[2(trimethylsilyl)ethoxy]methyl}-2,3-dihydro-l,3-benzothiazol-2-ylidene]amino}pyridazin3-yl)amino]-5-(piperidin-4-yl)-l,3-thiazole-4-carboxylate
Step A: bromo({1 -[(tert-butoxy)carbonyl]piperidin-4-yl])zinc
To a 50 mL Schlenk flask equipped with a magnetic stirrer bar was added 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 min with gentle stirring. While still under high vacuum the mixture was allowed to cool to ambient température, and was then swapped to the Schlenk line and back-filled with nitrogen. Tetrahydrofuran (15 mL) was added followed by 10 1,2-dibromoethane (0.06 mL, 0.75 mmol, 0.04 eq) and the mixture was heated to 60 °C over 10 min. The reaction was then removed from the heat and trimethylchlorosilane (0.02 mL, 0.15 mmol, 0.01 eq) was added, followed by iodine (0.5M in tetrahydrofuran; 0.15 mL, 0.175 mmol, 0.05 eq). The mixture was then heated at 60 °C for 10 min before cooling to < 35 °C. A solution of terLbutyl 4-bromopiperidine-l-carboxylate (5.28 g, 20 mmol, 1 eq) in tetrahydrofuran (10 mL) was added over 3 min and the mixture was then stirred at 50 °C for 6 h, then at ambient température overnight. The reaction was allowed to cool to ambient température then cannulation through a filter (cotton-wool/ celite / cotton-wool) under slight vacuum into a dry 25 mL Schlenk tube afforded the desired product as a 0.5M solution (as determined by titration with a 0.5M solution of iodine) that was used without further characterisation.
Step B: tert-butyl 4-[4-(ethoxycarbonyl)-2-[methyl(5-methyl-6-{[(2Z)-3-{[2(trimethylsilyl)ethoxy]methyl}-2,3-dihydro-l,3-benzothiazol-2-ylidene]amino}pyridazin-3yl) amino]-1,3-thiazol-5-yl]piperidine-l-carboxylate
To an oven dried flask was added the product from Préparation 11b (500 mg, 0.79 mmol, 1 eq) and Copper(I) lodide (300 mg, 1.57 mmol, 2 eq) and the mixture was evacuated and purged 25 with nitrogen (x3). Dimethylacetamide (15 mL) was added, resulting in a brown suspension that was allowed to stir for 5 mins. The product from Step A (0.5M in tetrahydrofuran; 9.44 mL, 4.72 mmol, 6 eq) was added and the mixture was stirred at ambient température overnight. The reaction was quenched by the addition of saturated aqueous ammonium chloride (25 mL), then further diluted with 1:1 water / saturated aqueous ammonium chloride (150 mL). The 30 organics were extracted with dichloromethane (x3) and the combined organic extracts were washed with brine, dried (magnésium sulfate), and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 40 g RediSep™ silica cartridge)
127 eluting with a gradient of 0 — 40% ethyl acetate in Ao-heptane afforded the desired product as a pale yellow foam (429 mg, 0.58 mmol, 74%).
LC/MS (C35H49N7O5SiS2) 740 [M+H]+; RT 1.40 (LCMS-V-B2)
Ή NMR (400 MHz, CDC13) 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-{[(2Z)-3-{[2-(trimethylsilyl)ethoxy]methyl}-2,3-dihydro1,3-benzothiazol-2-ylidene]amino}pyridazin-3-yl)amino]-5-(piperidin-4-yl)-l ,3-thiazole-4carboxylate
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 was quenched by the addition of saturated aqueous sodium bicarbonate, extracted with dichloromethane, and the organic extract washed with brine, dried (magnésium sulfate), and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 12 g RediSep™ silica cartridge) eluting with a gradient of 0 - 10% methanol in dichloromethane afforded the desired product as an off-white glass (59 mg, 0.09 mmol, 68%).
LC/MS (C3oH4iN703SiS2) 640 [M+H]+; RT 1.23 (LCMS-V-B1)
Ή NMR (400 MHz, DMSO-d6) 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).
Préparation 51: Ethyl 5-(azetidin-3-yl)-2-[methyl(5-methyl-6-{[(2Z)-3-{[2(trimethylsilyl)ethoxy]methyl}-2,3-dihydro-l,3-benzothiazol-2-ylidene]amino}pyridazin3-yl)amino]-l,3-thiazole-4-carboxylate
Step A: ethyl5-{l-[(tert-butoxy)carbonyl]azetidin-3-yl}-2-[methyl(5-methyl-6-{[(2Z)-3-{[2(trimethylsilyl)ethoxy]methyl}-2,3-dihydro-l,3-benzothiazol-2-ylidene]amino}pyridazin-3yl)amino]-l,3-thiazole-4-carboxylate
128
P To a solution of the product from Préparation 4a (401 mg, 0.98 mmol, 2 eq) in 1,4-dioxane (5 mL) was added the product from Préparation 3zb (168 mg, 0.49 mmol, 1 eq), N,Ndiisopropylethylamine (0.24 mL, 1.48 mmol, 3 eq), césium carbonate (481 mg, 1.48 mmol, 3 eq) and Xantphos (56.9 mg, 0.1 mmol, 0.2 eq) and the mixture was sparged with nitrogen (10 5 min). Tris(dibenzylideneacetone)dipalladium(0) (45.1 mg, 0.05 mmol, 0.1 eq) was added and the mixture was heated at 120 °C for 2 h under microwave irradiation. The reaction was diluted with dichloromethane then washed with water followed by brine, dried (magnésium sulfate), and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 24 g RediSep™ silica cartridge) eluting with a gradient of 0 - 100% ethyl 10 acetate in Ao-heptane afforded the desired product as a brown solid (99 mg, 0.14 mmol, 28%).
LC/MS (C33H45N7O5SiS2) 712 [M+H]+; RT 1.52 (LCMS-V-B1) XH NMR (400 MHz, DMSO-d6) 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 15 (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-{[(2Z)-3-{[2(trimethylsilyl)ethoxy]methyl}-2,3-dihydro-l,3-benzothiazol-2-ylidene]amino}pyridazin-3yl)amino]-l,3-thiazole-4-carboxylate
To a solution of the product from Step A (78 mg, 0.11 mmol, 1 eq) in dichloromethane (6 mL), 20 cooled in an ice-bath, was slowly added trifluoroacetic acid (0.5 mL, 6.71 mmol, 61.2 eq) slowly and the mixture was stirred at 0 °C for 5 h. The reaction was quenched by the addition of saturated aqueous sodium bicarbonate, then extracted with dichloromethane, washed with brine, dried (magnésium sulfate), and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 12 g RediSep™ silica cartridge) eluting with a 25 gradient of 0 - 10% methanol in dichloromethane afforded the desired product as a cream solid (29.6 mg, 0.05 mmol, 44%).
LC/MS (C28H37N7O3SiS2) 612 [M+H]+; RT 1.18 (LCMS-V-B1)
Ή NMR (400 MHz, DMSO-d6) 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 30 = 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).
129
Préparation 5m: Ethyl 5-|(lE)-3diydroxv-2-methylprop-1-en-l-yl|-2-|methyl(5methyl-6-{[(2Z)-3-{[2-(trimethylsilyl)ethoxy]methyl}-2,3-dihydro-l,3-benzothiazol-2ylidene]amino}pyridazin-3-yl)amino]-l,3-thiazole-4-carboxylate
Step A : tert-butyldimethyl(prop-2-yn-l -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 terAbutyldimethylsilyl chloride (20.2 g, 134 mmol, 1.5 eq) and the mixture was stirred at 0 °C for 90 min. The reaction was partitioned between dichloromethane and saturated aqueous ammonium chloride, separated (PTFE phase separator), and the organic phase concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 220 g RediSep™ silica cartridge) eluting with a gradient of 0 - 10% ethyl acetate in zso-heptane afforded the desired product as a clear oil (4.07 g, 23.9 mmol, 27%).
Ή NMR (400 MHz, DMSO-d6) 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-l,3,2-dioxaborolan-2yl)prop-2-en-l -yl]oxy}silane
To an oven-dried flask was added copper(I) chloride (237 mg, 2.39 mmol, 0.1 eq), Xantphos (1.38 g, 2.39 mmol, 0.1 eq), bis(pinacolato)diboron (6.67 g, 26.3 mmol, 1.1 eq) and tetrahydrofuran (17 mL) and the mixture was sparged 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 température overnight. The reaction was partitioned between ethyl acetate and water, and the organic phase was dried (magnésium sulfate), and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 80 g RediSep™ silica cartridge) eluting with a gradient of 0-5% ethyl acetate in Ao-heptane afforded the desired product as a clear oil (616 mg, 1.97 mmol, 8%).
Ή NMR (400 MHz, DMSO-d6) 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-[(IE)-3-[(tert-butyldimethylsilyl)oxy]-2-methylprop-l-en-1 -yl]-2-[methyl(5methyl-6-{[(2Z)-3-{[2-(trimethylsilyl)ethoxy]methyl}-2,3-dihydro-l,3-benzothiazol-2ylidene]amino}pyridazin-3-yl)amino]-l,3-thiazole-4-carboxylate
130
To a solution of the product from Préparation lie (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 water (2.5 mL) and potassium carbonate (136 mg, 0.98 mmol, 3 eq). The mixture was sparged with nitrogen (10 min) before adding [l,l'-bis(diphenylphosphino)ferrocene] dichloropalladium(II) (24 mg, 0.03 mmol, 0.1 eq) and the mixture was heated at 100 °C for 1 h under microwave irradiation. The reaction was partitioned between ethyl acetate and brine, and the organic phase was dried (magnésium sulfate) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 12 g RediSep™ silica cartridge) eluting with a gradient of 0 - 40% ethyl acetate in Ao-heptane afforded the desired product as a white solid (192 mg, 0.26 mmol, 79%).
LC/MS (C35H52N6O4Si2S2) 741 [M+H]+; RT 1.58 (LCMS-V-B2)
Ή NMR (400 MHz, DMSO-d6) 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-[(lE)-3-hydroxy-2-methylprop-l-en-l-yl]-2-[methyl(5-methyl-6-{[(2Z)-3{[2-(trimethylsilyl)ethoxy]methyl}-2,3-dihydro-l,3-benzothiazol-2-ylidene]amino}pyridazin3-yl)amino]-l,3-thiazole-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 (IM in tetrahydrofuran; 0.39 mL, 0.39 mmol, 1.5 eq) and the mixture was stirred at ambient température for 2 h. The reaction was partitioned between ethyl acetate and water, and the organic phase was dried (magnésium sulfate) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 4 g RediSep™ silica cartridge) eluting with a gradient of 0 - 60% ethyl acetate in isoheptane afforded the desired product as a yellow solid (160 mg, 0.26 mmol, 99%).
LC/MS (C29H38N6O4SiS2) 627 [M+H]+; RT 2.71 (LCMS-V-C)
Hl NMR (400 MHz, DMSO-d6) 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).
131
Préparation 6a: tert-Butyl 2V-[3-(3-fluoro-4-hydroxy-phenyl)prop-2-ynyl]-/V-methylcarbamate
Using Sonogashira General Procedure starting from 10.00 g of 2-fluoro-4-iodo-phenol (42.0 mmol, 1 eq.) as the appropriate phénol and 10.67 g of tert-butyl TV-methyl-A-prop-2-ynylcarbamate (63.1 mmol, 1.5 eq.) as alkyne reactant, 10.8 g (92%) of the desired product was obtained.
Ή NMR (500 MHz, DMSO-d6) δ ppm 10.32 (s, 1 H), 7.22 (brd, 1H), 7.08 (dm, 1H), 6.92 (dd, 1H), 4.21 (s, 2H), 2.85 (s, 3H), 1.41 (s, 9H); 13C NMR (125 MHz, DMSO-d6) 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-ESI (m/z): [M-C4H8+H]+ calcd for CnHuFNO3: 224.0717, found 224.0720.
Préparation 6b: 4-[3-(Dimethylamino)prop-l-ynyl]-2-fluoro-phenol
Using Sonogashira General Procedure starting from 10.00 g of 2-fluoro-4-iodo-phenol (42.0 mmol, 1 eq.) as the appropriate phénol and 5.24 g of N,N-dimethylprop-2-yn-l-amine (63 mmol, 1.5 eq.) as alkyne reactant, 7.30 g (90%) of the desired product was obtained.
Ή NMR (500 MHz, DMSO-d6) δ ppm 7.20 (dd, 1H), 7.07 (dm, 1H), 6.91 (m, 1H), 3.39 (m, 2H), 2.21 (m, 3H); 13C NMR (125 MHz, DMSO-d6) δ ppm 150.9, 146.2, 128.9, 119.5, 118.4, 113.6,84.5,84.2,48.2, 44.3; HRMS-ESI (m/z): [M+H]+ calcd for C11H13FNO: 194.0976, found 194.0981.
Préparation 6c: 2-Fluoro-4-(3-pyrrolidin-l-ylprop-l-ynyl)phenol
Step A: 3-(3-fluoro-4-triisopropylsilyloxy-phenyl)prop-2-yn-l-ol
A 500 mL oven-dried, one-necked, round-bottomed flask equipped with a PZFE-coated magnetic stirring bar was charged with 4.76 g of 2-fluoro-4-iodo-phenol (20 mmol, 1 eq.) and 3.96 g of K2CO3 (40 mmol, 2 eq.) then 100 mL of dry MeCN was added. To the resulting mixture 5.13 mL of TIPSCl (4.62 g, 24 mmol, 1.2 eq.) was added dropwise near intensive stirring at rt. The resulting mixture was stirred at room température for 30 min, while the reaction reached complété conversion. The reaction mixture was filtered through a pad of Celite to remove the solid particles then to the filtrate 2.33 mL of prop-2-yn-l-ol (2.24 g, 40 mmol, 2 eq.) and 20 mL of DIPA were added and placed under a nitrogen atmosphère through a gas inlet. After addition of 702 mg of Pd(PPh3)2C12 (1 mmol, 0.05 eq.) and 190 mg of Cul (1 mmol, 0.05 eq.) the resulting mixture was stirred at room température for 30 min, while the reaction
132 reached complété conversion. Celite was added to the reaction mixture and the volatiles were removed under reduced pressure. Then it was purified via flash column chromatography using heptane and EtOAc as eluents to give 5.2 g (81%) of the desired product as brown oil.
Ή NMR (500 MHz, DMSO-d6) 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); 13C NMR (125 MHz, DMSO-d6) 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-ESI (m/z): [M+H]+ calcd for CisHzsFOzSi: 323.1837, found 323.1832.
Step B: [2-fluoro-4-(3-pyrrolidin-l-ylprop-1 -ynyl)phenoxy]-triisopropyl-silane
Using Alkylation with in situ generated iodine General Procedure starting from 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.), 130 mg (34%) of the desired product was obtained.
1H NMR (500 MHz, DMSO-d6) 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); 13C NMR (125 MHz, DMSO-dô) 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; HRMSESI (m/z): [M+H]+ calcd for C22H35FNOSi: 376.2466, found 376.2456.
Step C: 2-fluoro-4-(3-pyrrolidin-l-ylprop-1 -ynyl)phénol
A 4 mL oven-dried vial equipped with a PTFE-coated magnetic stirring bar was charged with 83 mg of the product from Step B (0.221 mmol, 1 eq.) and 1.1 mL of dry THF then 265 uL of TB AF (1 M in THF, 225.9 mg, 0.2652 mmol, 1.2 eq.) was added dropwise at rt. The resulting mixture was stirred at rt for 15 min, when the reaction reached complété conversion. The reaction mixture was quenched with the addition of 200 uL of cc. NH4CI then Celite was added to the reaction mixture and the volatiles were removed under reduced pressure. Then it was purified via flash column chromatography using DCM and MeOH (1.2% NH3) as eluents to give 43 mg (88%) of the desired product.
Ή NMR (500 MHz, DMSO-dô) 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, 4H), 1.70 (m, 4H); 13C NMR (125 MHz, DMSO-dô) δ ppm 151.1, 146.8,128.9,119.4,118.5,113.1, 84.8, 83.8, 52.3,43.4, 23.8; HRMS-ESI (m/z): [M+H]+ calcd for C13H15FNO: 220.1132, found 220.1129.
Préparation 6d: 2-FIuoro-4-[3-(l-piperidyl)prop-l-ynyl]phenol
Step A: [2-fluoro-4-[3-(l-piperidyl)prop-l-ynyl]phenoxy]-triisopropyl-silane
133
Using Alkylation with in situ generated iodine General Procedure starting from 322 mg of
Préparation 6c, Step A (1 mmol, 1 eq.) as the appropriate alcohol and 425 mg of piperidine (5 mmol, 5 eq.), 250 mg (64%) of the desired product was obtained.
Ή NMR (500 MHz, DMSO-d6) 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); 13C NMR (125 MHz, DMSO-dô) 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; HRMS-ESI (m/z): [M+H]+ calcd for C23H37FNOSi: 390.2622, found 390.2608.
Step B: 2-Fluoro-4-[3-(l-piperidyl)prop-l-ynyl]phenol
A 4 mL oven-dried vial equipped with a PTFE-coated magnetic stirring 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 698 uL of TBAF (1 M in THF, 0.69 mmol, 1 eq.) was added dropwise at rt. The resulting mixture was stirred at rt for 15 min, when the reaction reached complété conversion. The reaction mixture was quenched with the addition of 200 uL of cc. NH4CI then Celite was added to the reaction mixture and the volatiles were removed under reduced pressure. Then it was purified via flash column chromatography using DCM and MeOH (1.2% NH3) as eluents to give 143 mg (87%) of the desired product.
'H NMR (500 MHz, DMSO-d6) 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); 13C NMR (125 MHz, DMSO-d6) 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; HRMS-ESI (m/z): [M+H]+ calcd for C14H17FNO: 234.1289, found 234.1292.
Préparation 6e: 2-Fluoro-4-(3-morpholinoprop-l-ynyl)phenol
Step A : [2-fluoro-4-(3-morpholinoprop-l-ynyl)phenoxy]-triisopropyl-silane
Using Alkylation with in situ generated iodine General Procedure starting from 322 mg of Préparation 6c, Step A (1 mmol, 1 eq.) as the appropriate alcohol and 435 mg of morpholine (5 mmol, 5 eq.), 160 mg (41%) of the desired product was obtained.
JH NMR (500 MHz, DMSO-d6) 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); 13C NMR (125 MHz, DMSO-d6) 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; HRMS-ESI (m/z): [M+H]+ calcd for C22H35FNO2Si: 392.2415, found 392.2408.
134
Step B: 2-fluoro-4-(3-morpholinoprop-l-ynyl)phenol
A 4 mL oven-dried vial equipped with a PTFE-coateà magnetic stirring 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 then 525 uL of TBAF (1 M in THF, 0.52 mmol, 1 eq.) was added dropwise at rt. The resulting mixture was stirred at rt for 15 min, when the reaction reached complété conversion. The reaction mixture was quenched with the addition of 200 uL of cc. NH4CI, then Celite was added to the reaction mixture and the volatiles were removed under reduced pressure. It was purified via flash column chromatography using DCM and MeOH (1.2% NH3) as eluents to give 120 mg (91%) of the desired product.
’H NMR (400 MHz, DMSO-d6) 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); 13C NMR (125 MHz, DMSO-d6) δ ppm 150.8, 146.2,128.9,119.5,118.4,113.6, 84.6, 84.1, 66.5, 52.2, 47.5; HRMS-ESI (m/z): [M+H]+ calcd for C13H15FNO2: 236.1081, found 236.1082.
Préparation 6f: 4-[3-(DimethyIamino)but-l-ynyl]-2-fluoro-phenol
Step A: 4-(3-fluoro-4-triisopropylsilyloxy-phenyl)but-3-yn-2-ol
A 500 mL oven-dried, one-necked, round-bottomed flask equipped with a PTFE-coated magnetic stirring bar. It was charged with 4.76 g of 2-fluoro-4-iodo-phenol (20 mmol, 1 eq.) and 3.96 g of K2COs (40 mmol, 2 eq.) then 100 mL of dry MeCN was added. To the resulting mixture 5.13 mL of TIPSCl (4.62 g, 24 mmol, 1.2 eq.) was added dropwise near intensive stirring at rt. The resulting mixture was stirred at room température for 30 min, while the reaction reached complété conversion. The reaction mixture was filtered through a pad of Celite to remove the solid particles then to the filtrate 3.10 mL of but-3-yn-2-ol (2.81 g, 40 mmol, 2 eq.) and 20 mL of DIPA were added and placed under a nitrogen atmosphère through a gas inlet. After addition of 702 mg of Pd(PPh3)2C12 (1 mmol, 0.05 eq.) and 190 mg of Cul (1 mmol, 0.05 eq.) the resulting mixture was stirred at room température for 30 min, while the reaction reached complété conversion. Celite was added to the reaction mixture and the volatiles were removed under reduced pressure. Then it was purified via flash column chromatography using heptane and EtOAc as eluents to give 6.2 g (92%) of the desired product as yellow oil.
Ή NMR (400 MHz, DMSO-d6) 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); 13C NMR (100 MHz, DMSO-d6) 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; HRMS-ESI (m/z): [M+H]+ calcd for Ci9H30FO2Si: 337.1994, found 337.1994.
135
Step B: 4-(3-fluoro-4-triisopropylsilyloxy-phenyl)-NJN-dimethyl-but-3-yn-2-amine
Using Alkylation with in situ generated iodine General Procedure 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), 360 mg (50%) of the desired product was obtained.
’H NMR (500 MHz, DMSO-d6) 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); 13C NMR (500 MHz, dmso-d6) 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; HRMS-ESI (m/z): [M+H]+ calcd for C2iH35FNOSi: 364.2466, found 364.2470.
Step C: 4-[3-(dimethylamino)but-l-ynyl]-2-fluoro-phenol
A 4 mL oven-dried vial equipped with a ΡΤΈΕ-coated magnetic stirring 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 660 uL of TBAF (1 M in THF, 0.66 mmol, 1.1 eq.) was added dropwise at rt. The resulting mixture was stirred at rt for 15 min, when the reaction reached complété conversion. The reaction mixture was quenched with the addition of 200 uL of cc. NH4CI, then Celite was added to the reaction mixture and the volatiles were removed under reduced pressure. Then it was purified via flash column chromatography using DCM and MeOH (1.2% NH3) as eluents to give 80 mg (70%) of the desired product.
Préparation 6g and 6h: 2-Fluoro-4-(3-morphoIinobut-l-ynyI)phenoI (enantiomers 1 and 2)
Step A: [2-fluoro-4-(3-morpholinobut-l-ynyl)phenoxy]-triisopropyl-silane
Using Alkylation with in situ generated iodine General Procedure starting from 644 mg of Préparation 6f, Step A (2 mmol, 1 eq.) as the appropriate alcohol and 5 mL of morpholine (10 mmol, 5 eq., 2 M solution in MeOH), 370 mg (45%) of the desired product was obtained.
Ή NMR (500 MHz, DMSO-d6) 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); 13C NMR (500 MHz, dmso-d6) 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; HRMS-ESI (m/z): [M+H]+ calcd for C23H37FNO2Si: 406.2572, found 406.2579.
Step B: 2-fluoro-4-(3-morpholinobut-l-ynyl)phenol
136
A 4 mL oven-dried vial equipped with a PTFE-coated magnetic stirring 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 990 uL of TBAF (1 M in THF, 0.99 mmol, 1.1 eq.) was added dropwise at rt. The resulting mixture was stirred at rt for 15 min, when the reaction reached complété conversion. The reaction mixture was quenched with the addition of 200 uL of cc. NH4CI, then Celite was added to the reaction mixture and the volatiles were removed under reduced pressure. Then it was purified via flash column chromatography using DCM and MeOH (1.2% NH3) as eluents to give 150 mg (60%) of the desired product.
Enantiomers were separated via chiral chromatography. Column: AD-H, Eluents: heptane /EtOH + 0.1% DEA; the enantiomer eluting earlier was collected as Préparation 6g with 99.8% ee and the enantiomer eluting later was collected as Préparation 6h with 99.4% ee.
Préparation 6i: 4- [1- [(Dimethylamino)methy 1] -3-bicyclo [1.1.1} pentany 1] phénol
Step A: l-(4-methoxyphenyl)-N,N-dimethyl-bicyclo[l.l.l]pentane-3-carboxamide
300 mg ofl-(4-methoxyphenyl)bicyclo[l.l.l]pentane-3-carboxylic acid (1.38 mmol, 1 eq.) and 278 mg of N,N-diethylethanamine (2.75 mmol, 2 eq.) were mixed in EtOAc (3 mL/mmol) then 1312 mg of 2,4,6-tripropyl-l,3,5,2/^(5},4/^(5),6/^(5}-trioxatriphosphinane 2,4,6-trioxide (50w% in EtOAc, 2.06 mmol, 1.5 eq.) was added in one portion then stirred at rt for 40 min. After the reaction time 1.03 mL oîN-methylmethanamine (2 M in MeOH, 2.06 mmol, 1.5 eq.) was added and stirred at rt until full conversion was observed (30 min). Reaction mixture was diluted with DCM then washed with cc. NaHCOs then the organic phase was washed with cc. NaCl, dried over MgSÛ4, filtered, concentrated, dried in vacuo to give 346 mg (quant.) of the desired product as a solid with peach color.
Ή NMR (500 MHz, DMSO-î/6) 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); 13C NMR (125 MHz, DMSO-d6) 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-ESI (m/z): [M+H]+ calcd for C15H20NO2: 246.1488, found 246.1495.
Step B: l-[3-(4-methoxyphenyl)-l-bicyclo[l.1 .l]pentanyl]-Nfil-dimethyl-methanamine
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 LÎA1H4 (1 M in THF, 2.36 mmol, 2 eq.) was added under nitrogen atmosphère at ambient température then stirred until full conversion was achieved (ca. 1 h). The mixture was cooled to 0°C then quenched with cc. NH4CI. After quenching ~5 mL water and ~10 mL EtOAc were added and shaked well. 2 M HCl was added and the (acidic) water phase was separated
137 then the organic phase was extracted with further 2 M HCl. The combined water phases were made basic with 2 M NaOH and extracted with DCM. The combined organic phases were washed with brine, dried over MgSÜ4 and concentrated, dried in vacuo. 219 mg (80%) of the desired product was obtained as viscous oil.
XH NMR (500 MHz, DMSO-d6) 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); 13C NMR (125 MHz, DMSO-d6) δ ppm 158.4, 133.4, 127.4, 114, 60.8, 55.5, 52.9, 46.6, 41.9, 38.0; HRMS-ESI (m/z): [M+H]+ calcd for C15H22NO: 232.1696, found 232.1700.
Step C: 4-[1 -[(dimethylamino)methyl]-3-bicyclo[1.1.1 ]pentanyl]phénol mg of the product from Step B (0.22 mmol, 1 eq.) was dissolved in DCM (5 mL/mmol) then 0.65 mL of BBr3 (1 M in DCM, 0.65 mmol, 3 eq.) was added under nitrogen atmosphère at 0°C then stirred for 30 min at 0°C and at rt until full conversion was achieved (ca. 45 min). DCM was added then poured into NaHCO3 solution, stirred for a few minutes then made it neutral with cc. NH4CI. The organic phase was separated and washed with brine, dried over MgSCh and concentrated, dried in vacuo. 47 mg (quant.) of the crude desired product was obtained as viscous oil.
Ή 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); 13C NMR (100 MHz, CDCh) 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]+ calcd for C14H20NO: 218.2, found 218.2.
Préparation 6i: N,N-Dimethyl-3-(4-triisopropylsilyloxyphenyl)prop-2-yn-l-amine
500 mg of 4-iodophenol (2.27 mmol, 1 eq.) and 628 mg of K2CO3 (4.55 mmol, 2 eq.) were mixed in acetonitrile (5 mL/mmol) then 526 mg of chloro(triisopropyl)silane (2.73 mmol, 1.2 eq.) was added at rt and stirred for 1 h. The reaction mixture was filtered through a pad of celite then 236 mg oîN,N-dimethylprop-2-yn-l-amine (2.84 mmol, 1.25 eq.), 50 mg of Pd(PPh3)2Cl2 (0.11 mmol, 0.05 eq.), 22 mg of Cul (0.11 mmol, 0.05 eq.) and 2.27 mL oiN-isopropylpropan2-amine (1 mL/mmol) were added then stirred at 50°C for 3 h. The reaction mixture was concentrated to Celite and purified via flash column chromatography using heptane and EtOAc as eluents to give 449 mg (60%) of the desired product as a yellow oil.
Ή NMR (500 MHz, DMSO-d6) 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); 13C NMR (125 MHz, DMSO-î/6) 156,133.5,120.3,116, 85.1/84.5, 44.3, 18.3, 12.4; HRMS-ESI (m/z): [M+H]+ calcd for C2oH34NOSi: 332.2404, found 332.2405.
138
Préparation 6k: 2-Fluoro-4-{3-[(4-methoxyphenyl)methoxy]prop-l-yn-l-yl}phenol
Step A : 4-bromo-2-fluorophenyl acetate
To a solution of 4-bromo-2-fluorophenol (2.29 mL, 20.9 mmol, 1 eq) in dichloromethane (60 mL) was added 4-(dimethylamino)pyridine (5.12 g, 41.9 mmol, 2 eq) and acetic anhydride (4.94 mL, 52.4 mmol, 2.5 eq) and the mixture was stirred at ambient température for 18 h. The reaction 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 cartridge) eluting with a gradient of 0 - 10% ethyl acetate in zTo-heptane afforded the desired product as a clear oil (5.1 g, 21.9 mmol, > 100%).
Ή NMR (400 MHz, DMSO-d6) 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-l -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 l-(bromomethyl)-4-methoxybenzene (3.09 mL, 21.4 mmol, 1.2 eq) and the mixture was stirred at ambient température for 18 h. The reaction was partitioned between ethyl acetate and saturated aqueous ammonium chloride, and the organic phase was dried (magnésium sulfate), filtered and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 80 g RediSep™ silica cartridge) eluting with a gradient of 0 — 10% ethyl acetate in z’so-heptane afforded the desired product as a clear oil (2.44 g, 13.9 mmol, 78%).
Ή NMR (400 MHz, DMSO-d6) 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-l-yn-l-yl}phenyl acetate
To an oven-dried pressure tube was added the product from Step A (1.68 g, 7.23 mmol, 1 eq), triethylamine (18 mL), the product from Step B (1.91 g, 10.8 mmol, 1.5 eq), copper (I) 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 sparged with nitrogen (10 mins) before sealing and heating at 90 °C for 4 h. The reaction was allowed to cool to ambient température, then partitioned between dichloromethane and brine, and the organic phase was separated and dried (PTFE
139 phase separator) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 40 g RediSep™ silica cartridge) eluting with a gradient of 0 - 30% ethyl acetate in Ao-heptane afforded the desired product as an orange solid (2.11 g, 6.43 mmol, 89%).
Ή NMR (400 MHz, DMSO-d6) 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-l-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 température for 3 h. The reaction 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 cartridge) eluting with a gradient of 0 - 40% ethyl acetate in Ao-heptane afforded the desired product as a yellow gum (128 mg, 0.45 mmol, 73%).
LC/MS (C17H15FO3) 285 [M-H]-; RT 2.09 (LCMS-V-C)
Ή NMR (400 MHz, DMSO-d6) 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).
Préparation 7c: 3,6-DichIoro-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 1.6M aqueous ammonium persulfate (62.9 mL, 101 mmol, 3 eq) was added dropwise and the reaction was heated at 70 °C for 30 mins. The mixture was allowed to cool to ambient température, then cooled to 0 °C and neutralised with concentrated ammonium hydroxide solution slowly to pH 7 then adjusted to pH 8-9. A yellow precipitate formed which was filtered and washed with water and ethyl acetate to afford a white solid. The filtrâtes were combined, the layers separated, and the aqueous phase extracted with ethyl acetate (100 mL). The combined organics were washed with brine (100 mL), dried (magnésium sulfate) and concentrated in vacuo to obtain a yellow oil. Purification by automated flash column chromatography (CombiFlash Rf, 80 g RediSep™ silica cartridge) eluting with a gradient of 0
140
-5% ethyl acetate in heptane afforded the desired product as a colourless oil (4.61 g, 24.1 mmol, 72%).
LC/MS (C7H8CI2N2) 191 [M+H]+; RT 1.11 (LCMS-V-B1) XH NMR (400 MHz, DMSO-d6) δ 8.00 (d, J = 0.6 Hz, 1H), 3.16 (h, 1H), 1.26 (d, J = 6.8 Hz, 6H).
Préparation 7e: 3,6-Dichloro-4-methyl-5-(propan-2-yl)pyridazine
A suspension of 3,6-dichloro-4-methyl-pyridazine (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. 1.6M aqueous ammonium persulfate (5.75 mL, 9.2 mmol, 3 eq) was added dropwise and the mixture was stirred for 30 min, then allowed to cool to ambient température. Neutralisation with ammonium hydroxide, filtration through celite, and elution with ethyl acetate afforded a biphasic mixture. The layers were separated, the aqueous phase was extracted with ethyl acetate, and the combined organics were dried (magnésium sulfate) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 40 g RediSep™ silica cartridge) eluting with a gradient of 0 - 15% ethyl acetate in zso-heptane afforded the desired product as a white solid (476 mg, 2.32 mmol, 76%).
LC/MS (C8H10CI2N2) 205 [M+H]+; RT 1.15 (LCMS-V-B1) XH NMR (400 MHz, DMSO-d6) δ 3.57 (hept, J = 7.2 Hz, 1H), 2.49 (s, 3H), 1.35 (d, J = 7.2 Hz, 6H).
Préparation 7f: 3,6-Dichloro-4-ethyl-5-methylpyridazine
To a stirred suspension of 3,6-dichloro-4-methylpyridazine (5 g, 30.7 mmol, 1 eq) and propionic acid (2.75 mL, 36.8 mmol, 1.2 eq) in water (100 mL) was added 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 was heated to 60 °C. 1.6M aqueous ammonium persulfate (57.5 mL, 92 mmol, 3 eq) was then added dropwise over 20 min. The reaction was heated to 70 °C for 30 min and then left cool to ambient température. The mixture was basified to pH 8 using conc. ammonium hydroxide and the product extracted with ethyl acetate (100 mL x 2). The combined organic extracts were dried (magnésium sulfate) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 120 g
141 (φ RediSep™ silica cartridge) eluting with a gradient of 0 - 25% ethyl acetate in Ao-heptane afforded the desired product as a white solid (4.11 g, 21.5 mmol, 70%).
LC/MS (C7H8CI2N2) 191 [M+H]+; RT 2.17 (LCMS-V-C)
Ή NMR (400 MHz, DMSO-d6) δ 2.82 (q, J = 7.6 Hz, 2H), 2.44 (s, 3H), 1.13 (t, J = 7.6 Hz, 5 3H).
Préparation 7g: 4-[(Benzyloxy)methyl]-3,6-dichloro-5-methylpyridazine
To a suspension of 3,6-dichloro-4-methylpyridazine (5.1 g, 30.7 mmol, 1 eq) and benzyloxyacetic acid (5.61 g, 33.7 mmol, 1.1 eq) in water (100 mL) was added silver nitrate (5.21 g, 30.7 mmol, 1 eq) and the mixture was heated to 50 °C. Sulfuric acid (4.91 mL, 92 10 mmol, 3 eq) was added and the mixture was heated to 60 °C. 1.6M aqueous ammonium persulfate (57.5 mL, 92 mmol, 3 eq) was added dropwise and the reaction was heated at 70 °C for 30 min. The mixture was allowed to cool to ambient température and basified to pH 8 with conc. aqueous ammonium hydroxide. The mixture was extracted with ethyl acetate (2 x 150 mL) and the combined organics were washed with brine (200 mL), dried (magnésium sulfate), 15 and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 120 g RediSep™ silica cartridge) eluting with a gradient of 0 - 25% ethyl acetate in zso-heptane afforded the desired product as a yellow oil (4.67 g, 16.5 mmol, 54%).
LC/MS (C13H12CI2N2O) 283 [M+H]+; RT 1.26 (LCMS-V-B1)
Ή NMR (400 MHz, DMSO-d6) δ 7.42 - 7.27 (m, 5H), 4.71 (s, 2H), 4.60 (s, 2H), 2.46 (s, 20 3H).
Préparation 7h: 3,6-Dichloro-4-cyclopropyl-5-methylpyridazine
To a suspension of 3,6-dichloro-4-methylpyridazine (4.88 g, 29.9 mmol, 1 eq) and cyclopropanecarboxylic acid (2.63 mL, 32.9 mmol, 1.1 eq) in water (100 mL) was added silver nitrate (5.09 g, 29.9 mmol, 1 eq) and the mixture was heated to 50 °C. To this was added sulfuric 25 acid (4.79 mL, 89.8 mmol, 3 eq) and the mixture was heated to 60 °C. 1.6M aqueous ammonium persulfate (56.1 mL, 89.8 mmol, 3 eq) was added dropwise over 20 min then the mixture was heated to 70 °C and stirred for 30 min. The mixture was allowed to cool to ambient température and basified to pH 8 with conc. ammonium hydroxide. The product was extracted into the organic phase using ethyl acetate (2 x 200 mL) and the combined organic extracts were washed 30 with brine, dried (magnésium sulfate) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 120 g RediSep™ silica cartridge) eluting with
142 a gradient of 0 - 30% ethyl acetate in Ao-heptane afforded the desired product as a white solid (3.99 g, 19.7 mmol, 66%).
LC/MS (C8H8C12N2) 203 [M+H]+; RT 1.09 (LCMS-V-B1) XH NMR (400 MHz, DMSO-d6) 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).
Préparation 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. 1.6M aqueous ammonium persulfate (25.2 mL, 40.3 mmol, 3 eq) was added dropwise over 15 mins and the reaction was then heated at 70 °C for 30 min. The mixture was allowed to cool to ambient température and basified to pH 10 with conc. ammonium hydroxide. The product was extracted into ethyl acetate (2 x 150 mL) and the extracts were washed with brine, dried (magnésium sulfate) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 80 g RediSep™ silica cartridge) eluting with a gradient of 0 — 20% ethyl acetate in Ao-heptane afforded the desired product as a white solid (985 mg, 5.21 mmol, 39%).
LC/MS (C7H6CI2N2) 189 [M+H]+; RT 1.00 (LCMS-V-B1) XH NMR (400 MHz, DMSO-d6) 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).
Préparation 7m: 4-[3-(Benzyloxy)propyl]-3,6-dichloro-5-methylpyridazine
Step A: [(pent-4-yn-l-yloxy)methyl]benzene
To an oven-dried flask was added 4-Pentyn-l-ol (5.53 mL, 59.4 mmol, 1 eq) and tetrahydrofuran (50 mL) and the solution was cooled to 0 °C. Sodium hydride (60%; 2.85 g, 119 mmol, 2 eq) was added portionwise and the reaction 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 was allowed to warm to ambient température and stirred for 16 h. The reaction was cooled to 0 °C and quenched by the addition of saturated aqueous ammonium chloride (20 mL). Water (20 mL) was added and the layers separated. The aqueous layer was extracted with ethyl acetate (2 x 50 mL) and the combined organics were washed with brine (50 mL), dried (magnésium sulfate), and concentrated in vacuo. Purification by automated flash column chromatography
143 (φ (CombiFlash Rf, 120 g RediSep™ silica cartridge) eluting with a gradient of 0 - 10% ethyl acetate in zso-heptane afforded the desired product as a yellow oil (8.93 g, 51.3 mmol, 86%).
LC/MS (C12H14O) 175 [M+H]+; RT 1.25 (LCMS-V-B1)
Ή NMR (400 MHz, CDCI3) δ 7.42 - 7.21 (m, 5H), 4.52 (s, 2H), 3.57 (t, J = 6.2 Hz, 2H), 5 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-l-yloxy)methyl]benzene
To an oven-dried flask was added a solution of the product from Step A (3.9 g, 22.4 mmol, 1 eq) in tetrahydrofuran (30 mL) under nitrogen. The reaction was cooled to -78 °C and nbutyllithium (2.2M in hexane; 12.2 mL, 26.9 mmol, 1.2 eq) was added over 30 min and the 10 mixture was stirred at -78 °C for 1 h. lodomethane (2.09 mL, 33.6 mmol, 1.5 eq) was added dropwise and the résultant mixture was allowed to warm to 0 °C over 2 h. The reaction was quenched by the addition of saturated aqueous saturated 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 15 mL), dried (magnésium sulfate) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 80 g RediSep™ silica cartridge) eluting with a gradient of 0- 10% ethyl acetate in zso-heptane afforded the desired product as a straw-coloured liquid (3.61 g, 19.2 mmol, 86%).
LC/MS (C13H16O) no ionisation; RT 1.15 (LCMS-V-B1)
Ή NMR (400 MHz, DMSO-d6) 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-l,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 h. 25 The mixture was allowed to cool to ambient température and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 80 g RediSep™ silica cartridge) eluting with a gradient of 0 - 35% ethyl acetate in iso-heptane afforded the desired product as an orange oil (3.06 g, 9.83 mmol, 62%).
LC/MS (C15H16CI2N2O) 311 [M+H]+; RT 1.35 (LCMS-V-B1)
Ή NMR (400 MHz, DMSO-d6) 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).
144
Préparation 8ca and 8cb: 6-Chloro-5-(propan-2-yl)pyridazin-3-amine and 6-Chloro-4(propan-2-yl)pyridazin-3-amine
The product from Préparation 7c (3.61 g, 18.9 mmol, 1 eq) was suspended in ammonium hydroxide (20 mL) and heated at 150 °C for 4 h under microwave irradiation. The solids were 5 collected by filtration, washed with water and dried to afford 6-chloro-5-(propan-2yl)pyridazin-3-amine (2.06 g, 12 mmol, 64%) as an off-white solid [~ 9:1 mix with other regioisomer] (Préparation 8ca).
LC/MS (C7H10CIN3) 172 [M+H]+; RT 0.81 (LCMS-V-B1) *H NMR (400 MHz, DMSO-d6) δ 6.76 (d, J = 0.7 Hz, 1H), 6.51 (s, 2H), 3.06 - 2.82 (m, 1H), 10 1.18 (d, 6H).
The filtrate was concentrated in vacuo and the crude solid residue was extracted with dichloromethane, filtered, and evaporated to afford 6-chloro-4-isopropylpyridazine-3-amine (447 mg, 2.6 mmol, 14%) as a yellow oil [~ 7:3 mix with other regioisomer] (Préparation 8cb).
LC/MS (C7H10CIN3) 172 [M+H]+; RT 0.79 (LCMS-V-B1)
Hl NMR (400 MHz, DMSO-d6) δ 7.23 (s, 1H), 6.53 (s, 2H), 2.95 - 2.80 (m, 1H), 1.15 (d, 6H).
Préparation 8d: 6-Chloro-5-methylpyridazin-3-amine and 6-Chloro-4methylpyridazin-3-amine
A suspension of 3,6-dichloro-4-methyl-pyridazine (2 g, 12.3 mmol, 1 eq) in ammonium 20 hydroxide (40 mL) was heated in a pressure reactor at 150 °C for 6 h, then allowed to cool to ambient température. The résultant precipitate was filtered, washed with water (10 mL) and dried to afford the desired product as fine brown needles (1.31 g, 0.01 mol, 74%) [~ 6:4 ratio of regioisomers], The mixture of isomers was used in the next step without further purification.
LC/MS (C5H6CIN3) no ionisation; RT 0.28 (LCMS-V-B1)
Préparation 8e: 6-Chloro-4-methyl-5-(propan-2-yl)pyridazin-3-amine and 6Chloro-5-methyl-4-(propan-2-yl)pyridazin-3-amine
The product from Préparation 7e (480 mg, 2.34 mmol, 1 eq) was introduced into the ‘bomb’ apparatus. Ammonium hydroxide (15 mL) was added, the vessel sealed and the mixture was stirred at 160 °C for 6 h. After allowing to cool to ambient température, the vessel was opened
145 and the reaction mixture was concentrated in vacuo and the mixture of isomers was used in the next step without purification.
LC/MS (C8H12CIN3) 186 [M+H]+; RT 0.88 (LCMS-V-B1)
Préparation 8f: 6-ChIoro-5-ethyl-4-methyIpyridazin-3-amine and 6-Chloro-4-ethyl5-methylpyridazin-3-amine
A suspension of the product from Préparation 7f (4 g, 21 mmol, 1 eq) in ammonium hydroxide (15 mL) was stirred at 160 °C in a sealed flask for 24 h. The mixture was allowed to cool to ambient température and a brown solid precipitated. The solid was collected by filtration, washed with water, and dried under vacuum to afford 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 (C7H10CIN3) 172 [M+H]+; RT 1.53 (LCMS-V-C)
Préparation 8g: 5-[(Benzyloxy)methyl]-6-chloro-4-methylpyridazin-3-amine and 4[(Benzyloxy)methyl]-6-chloro-5-methylpyridazin-3-amine
The product from Préparation 7g (600 mg, 2.12 mmol, 1 eq) was suspended as an oil in ammonium hydroxide (7 mL) and heated at 160 °C for 5 h in a sealed tube. The mixture was allowed to cool to ambient température and a brown solid precipitated. The solid was collected by filtration, washed with water and dried under vacuum to afford the desired product (455 mg, 1.73 mmol, 81%) as a mixture of two regioisomers that was used directly in the next step.
LC/MS (C13H14CIN3O) 264 [M+H]+; RT 1.02 (LCMS-V-B1)
Préparation 8ha and 8hb: 6-Chloro-5-cycIopropyl-4-methylpyridazin-3-amine and 6Chloro-4-cyclopropyI-5-methylpyridazin-3-amine
The product from Préparation 7h (3.99 g, 19.7 mmol, 1 eq) was suspended in ammonium hydroxide (10 mL) and heated in a sealed tube for 20 h. The mixture was allowed to cool to ambient température and the precipitated solid was collected by filtration and dried to afford 6chloro-5-cyclopropyl-4-methylpyridazin-3-amine (2.44 g, 13.3 mmol, 68%) [Préparation 8ha].which was used directly in the next step without further purification.
LC/MS (C8HioC1N3) 184 [M+H]+; RT 0.80 (LCMS-V-B1)
The filtrate was concentrated under reduced pressure, then evaporated from toluene (x 3) to remove residual water, to afford crude material (2.00 g, 10.89 mmol, 55%) containing 6-chloro20487
146
4-cyclopropyl-5-methylpyridazin-3-amine that was used directly in subséquent steps without further purification [Préparation 8hb].
LC/MS (C8HioC1N3) 184 [M+H]+; RT 0.79 (LCMS-V-B1)
Préparation 8ia and 8ib: (6-Amino-3-chloro-5-methylpyridazin-4-yl)methanol and (3Amino-6-chloro-5-methylpyridazin-4-yl)methanol
To a cooled solution of the product from Préparation 8g (3.1 g, 11.8 mmol, 1 eq) in dichloromethane (100 mL) was added boron trichloride solution (58.8 mL, 1 M, 0.06 mol, 5 eq) dropwise and the mixture was stirred at ambient température for 2 h. The reaction was cooled to 0 - 5 °C and cautiously quenched by the addition of methanol (30 mL). Further methanol (30 mL) was added and the mixture was heated at reflux for 30 min. The solvent was removed in vacuo and purification by automated flash column chromatography (CombiFlash Rf, 120 g RediSep™ silica cartridge) eluting with a gradient of 0 - 17.5% methanol in dichloromethane afforded the desired regioisomeric products:
(3-Amino-6-chloro-5-methylpyridazin-4-yl)methanol as a beige solid (0.87 g, 5.0 mmol, 42%) [Préparation 8ib].
LC/MS (C6H8C1N3O) 174 [M+H]+; RT 0.26 (LCMS-V-B1)
Hl NMR (400 MHz, DMSO-d6) 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%) [Préparation 8ia].
LC/MS (C6H8C1N3O) 174 [M+H]+; RT 0.18 (LCMS-V-B1) ’H NMR (400 MHz, DMSO-d6) δ 8.18 (s, 2H), 4.56 (s, 2H), 2.31 (s, 3H).
Préparation 8ia: 6-Chloro-5-(methoxymethyI)-4-methylpyridazin-3-amine
Step A: 5-(bromomethyl)-6-chloro-4-methylpyridazin-3-amine
To a suspension of the product from Préparation 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 température. The reaction was quenched by addition of methanol (20 mL) and concentrated in vacuo to afford a solid (3.2 g). The material was used directly in the next step without further purification.
LC/MS (C6H7BrClN3) 238 [M+H]+; RT 0.81 (LCMS-V-B1)
Step B: 6-chloro-5-(methoxymethyl)-4-methylpyridazin-3-amine
147
A solution of the crude product from Step A (780 mg) in methanol (10 mL) was heated at 120
C for 1 h under microwave irradiation. The mixture was concentrated in vacuo and purification by automated flash column chromatography (CombiFlash Rf, 12 g RediSep™ silica cartridge) eluting with a gradient of 0 - 10% methanol in dichloromethane afforded the desired product as a brown oil (323 mg, 0.90 mmol, 66% over both steps).
LC/MS (C7H10CIN3O) 188 [M+H]+; RT 0.61 (LCMS-V-B1)
Préparation 8jb: 6-Chloro-4-(methoxymethyl)-5-methylpyridazin-3-amine
Step A: 4-(bromomethyl)-6-chloro-5-methylpyridazin-3-amine
To a suspension of the product from Préparation 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 température. The reaction was quenched by the addition of methanol (20 mL) and then the volatile organics were removed in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 24 g RediSep™ silica cartridge) eluting with a gradient of 0 — 5% methanol in dichloromethane afforded the desired product as an orange solid (695 mg, 2.94 mmol, 100%).
LC/MS (C6H7BrClN3) 238 [M+H]+; RT 0.82 (LCMS-V-B1) ’H NMR (400 MHz, DMSO-d6) 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 purification by automated flash column chromatography (CombiFlash Rf, 24 g RediSep™ silica cartridge) eluting with a gradient of 0 - 5% methanol in dichloromethane afforded the desired product as a purple crystalline solid (91 mg, 0.49 mmol, 27%).
LC/MS (C7HioC1N30) 188 [M+H]+; RT 0.70 (LCMS-V-B1)
Ή NMR (400 MHz, DMSO-d6) δ 7.66 (s, 2H), 4.50 (s, 2H), 3.32 (s, 3H), 2.38 (s, 3H).
Préparation 8ka: 6-Chloro-5-(ethoxymethyl)-4-methylpyridazin-3-amine
A solution of the product from Préparation 8ja, Step A (500 mg, 2.11 mmol, 1 eq) in éthanol (15 mL)was heated at 120 °C for 1 h under microwave irradiation. The mixture was concentrated in vacuo and purification by automated flash column chromatography
148 (CombiFlash Rf, 24 g RediSep™ silica cartridge) eluting with a gradient of 0 - 10% methanol in dichloromethane afforded the desired product as a thick oil that crystallised on scratching (136 mg, 0.67 mmol, 32%).
LC/MS (C8H12CIN3O) 202 [M+H]+; RT 0.63 (LCMS-V-B1) ‘H NMR (400 MHz, DMSO-d6) 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).
Préparation 8ma and 8mb: 5-[3-(Benzyloxy)propyl]-6-chIoro-4-methylpyridazin-3amine and 4-[3-(Benzyloxy)propyl]-6-chloro-5-methylpyridazin-3-amine
The product from Préparation 7m (3.06 g, 9.83 mmol, 1 eq) and ammonium hydroxide (15 mL) 10 were heated in a sealed flask for 48 h at 160 °C. The mixture was allowed to cool to ambient température and extracted with dichloromethane (2 x 100 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 cartridge) eluting with a gradient of 0 - 100% ethyl acetate in Ao-heptane afforded the desired regioisomeric products:
5-[3-(Benzyloxy)propyl]-6-chloro-4-methylpyridazin-3-amine as a beige solid (1.3 g, 4.46 mmol, 45%) [Préparation 8ma].
LC/MS (C15H18CIN3O) 292 [M+H]+; RT 1.12 (LCMS-V-B1) XH NMR (400 MHz, DMSO-d6) 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-(BenzyIoxy)propyl]-6-chloro-5-methylpyridazin-3-amine as a beige solid (500 mg, 1.71 mmol, 17%) [Préparation 8mb].
LC/MS (C15H18CIN3O) 292 [M+H]+; RT 1.15 (LCMS-V-B1) XH NMR (400 MHz, DMSO-d6) 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).
Préparation 9a: A-(6-Chloro-4,5-dimethylpyridazin-3-yl)-l,3-benzoihiazol-2-amine
To a solution of 3-amino-6-chloro-4,5-dimethylpyridazine (450 mg, 2.86 mmol, 1 eq) and 2bromo-l,3-benzothiazole (672 mg, 3.14 mmol, 1.1 eq) in 1,4-dioxane (20 mL) was added sodium hydride (60% dispersion; 457 mg, 11.42 mmol, 4 eq) portionwise and the mixture was refluxed for 16 h. The reaction mixture was allowed to cool to ambient température and then 30 quenched with acetic acid. The mixture was diluted with ethyl acetate (200 mL) and washed
149 with aqueous saturated sodium bicarbonate (100 mL), and brine (100 mL). The organic extract was dried (magnésium sulfate) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 40 g RediSep™ silica cartridge) eluting with a gradient of 0 - 3% methanol in dichloromethane afforded the desired product as a yellow solid (673 mg, 2.32 mmol, 81%).
LC/MS (C13H11CIN4S) 291 [M+H]+; RT 1.22 (LCMS-V-B1) XH NMR (400 MHz, DMSO-d6) 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).
Préparation 9b: N-(6-Chloropyridazin-3-yl)-l,3-benzothiazol-2-amine
To a solution of 3-amino-6-chloropyridazine (2 g, 15.4 mmol, 1 eq) and 2-bromo-l,3benzothiazole (3.64 g, 17.0 mmol, 1.1 eq) in 1,4-dioxane (120 mL) was added sodium hydride (60% dispersion; 2.47 g, 61.8 mmol, 4 eq) portionwise and the mixture was refluxed for 30 mins. The reaction mixture was allowed to cool to ambient température, and was then cooled in an ice-water bath and quenched with methanol. Removal of the methanol by rotary évaporation caused a solid to precipitate. The mixture was cooled in ice-water and the solid was collected by filtration, washed with ice-cold 1,4-dioxane (100 mL), 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 afford the desired product (1.94 g, 7.38 mmol, 48%).
LC/MS (C11H7CIN4S) 263 [M+H]+; RT 1.1 (LCMS-V-B1) >H NMR (400 MHz, DMSO-d6) 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).
Préparation 9ca: A-[6-Chloro-5-(propan-2-yl)pyridazin-3-yl]-l,3-benzothiazol-2amine
To a suspension of the product from Préparation 8ca (2.06 g, 12 mmol, 1 eq) in 1,4-dioxane (70 mL) was added 2-bromo-l,3-benzothiazole (2.83 g, 13.2 mmol, 1.1 eq) followed by sodium hydride (60% dispersion; 0.96 g, 24.01 mmol, 2 eq) portionwise and the mixture was heated at 90 °C for 30 min. After allowing to cool to ambient température the reaction was cooled in icewater, quenched by the addition of acetic acid, and concentrated in vacuo. The résultant solid
150 was triturated with water, filtered, and dried under vacuum. Purification by automated flash column chromatography (CombiFlash Rf, 120 g RediSep™ silica cartridge) eluting with a gradient 0 - 50% ethyl acetate in Ao-heptane afforded a solid that was triturated with ethyl acetate, filtered, and dried under vacuum to afford the desired product as an off-white solid 5 (1.54 g, 5.05 mmol, 42%).
LC/MS (C14H13CIN4S) 305 [M+H]+; RT 1.31 (LCMS-V-B1)
Ή NMR (400 MHz, DMSO-d6) 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).
Préparation 9cb: A^-[6-Chloro-4-(propan-2-yl)pyridazin-3-yI]-l,3-benzothiazol-2amine
To a solution of the product from Préparation 8cb (47 mg, 2.6 mmol, 1 eq) and 2-bromo-l,3benzothiazole (669 mg, 3.13 mmol, 1.2 eq) in 1,4-dioxane (10 mL) was added sodium hydride (60% dispersion; 208 mg, 5.21 mmol, 2 eq) portionwise and the mixture was refluxed for 1 h. 15 The mixture was allowed to cool to ambient température then cooled to 0 °C and quenched by addition of acetic acid. After évaporation the residue was partitioned between ethyl acetate and water and the organic phase was washed with brine, dried (magnésium sulfate) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 24 g RediSep™ silica cartridge) eluting with a gradient of 0 — 20% ethyl acetate in Ao-heptane 20 afforded the desired product as a bright yellow solid (367 mg, 1.2 mmol, 46%).
LC/MS (C14H13CIN4S) 305 [M+H]+; RT 1.35 (LCMS-V-B1) ‘H NMR (400 MHz, DMSO-d6) 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).
Préparation 9da and 9db: /V-(6-Chloro-5-methylpyridazin-3-yl)-l,3-benzothiazol-225 amine and 7V-(6-Chloro-4-methylpyridazin-3-yl)-l,3-benzothiazol-2-amine
To a suspension of the product from Préparation 8d (1.31 g, 9.1 mmol, 1 eq) and 2-bromo-l,3benzothiazole (2.14 g, 10.01 mmol, 1.1 eq) in 1,4-dioxane (50 mL) was added sodium hydride (60% dispersion; 0.73 g, 18.2 mmol, 2 eq) portionwise and the mixture was refluxed for 1 h. The reaction mixture was allowed to cool to ambient température, quenched with acetic acid, 30 then neutralised with aqueous sodium bicarbonate. The mixture was extracted with ethyl acetate (100 mL x3) and the combined organic extracts were washed with brine, dried (magnésium
151 sulfate), and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 120 g RediSep™ silica cartridge) eluting wiht a gradient of 0 — 80% ethyl acetate in Ao-heptane afforded the desired products:
7V-(6-Chloro-4-methylpyrîdazin-3-yl)-l,3-benzothiazol-2-amine as a yellow solid (140 mg, 0.51 mmol, 5.6%) [Préparation 9db].
LC/MS (C12H9CIN4S) 277 [M+H]+; RT 1.18 (LCMS-V-B1)
Ή NMR (400 MHz, DMSO-d6) 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).
N-(6-Chloro-5-methylpyridazin-3-yl)-l,3-benzothiazol-2-amine as a yellow solid (69 mg, 0.25 mmol, 2.7%) [Préparation 9da],
LC/MS (C12H9CIN4S) 277 [M+H]+; RT 1.15 (LCMS-V-B1) 1H NMR (400 MHz, DMSO-d6) 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).
Préparation 9ea and 9eb: \-[6-Chloro-4-methyl-5-(propan-2-yl)pyridazin-3-yl|-1.3benzothiazol-2-amine amZ7V-[6-Chloro-5-methyl-4-(propan-2-yl)pyridazin-3-yl]-l,3benzothiazol-2-amine
To a suspension of the product from Préparation 8e (434 mg, 2.34 mmol, 1 eq) and 2-bromo1,3-benzothiazole (551 mg, 2.57 mmol, 1.1 eq) in 1,4-dioxane (10 mL) was added sodium hydride (60% dispersion; 187 mg, 4.68 mmol, 2 eq) portionwise and the mixture was refluxed for 1 h. The reaction mixture was allowed to cool to ambient température and then quenched with acetic acid. The mixture was then diluted with aqueous sodium bicarbonate and the product extracted with dichloromethane. The organics were washed with brine, dried (magnésium sulfate) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 40 g RediSep™ silica cartridge) eluting with a gradient of 0 - 30% ethyl acetate in Ao-heptane afforded the desired product regioisomers:
7V-[6-Chloro-5-methyI-4-(propan-2-yl)pyridazin-3-yl]-l,3-benzothiazol-2- amine as a yellow solid (114 mg, 0.36 mmol, 15%) [Préparation 9eb].
LC/MS (C15H15CIN4S) 319 [M+H]+; RT 1.39 (LCMS-V-B1)
152 φ XH NMR (400 MHz, DMSO-d6) 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).
N-[6-Chloro-4-methyl-5-(propan-2-yl)pyridazin-3-yl]-l,3-benzothiazol-2-amine as an 5 orange solid (300 mg, 0.94 mmol, 40%) [Préparation 9ea].
LC/MS (C15H15CIN4S) 319 [M+H]+; RT 1.36 (LCMS-V-B1)
Ή NMR (400 MHz, DMSO-d6) 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).
Préparation 9fa and 9fb: A-(6-Chloro-5-ethyl-4-methylpyridazin-3-yl)-l,310 benzothiazol-2-amine and A-(6-Chloro-4-ethyl-5-methylpyridazin-3-yl)-l,3-benzothiazol2-amine
To a suspension of the product from Préparation 8f (1.4 g, 8.16 mmol, 1 eq) and 2-bromo-l,3benzothiazole (2.1 g, 9.79 mmol, 1.2 eq) in 1,4-dioxane (50 mL) was added sodium hydride (60% dispersion; 1.31 g, 32.6 mmol, 4 eq) portionwise. The mixture was heated at reflux for 15 1.5 h then allowed to cool to ambient température. The reaction was quenched with acetic acid and neutralised with sodium bicarbonate. The mixture was extracted with ethyl acetate (x2), and the combined organic extracts were washed with brine, dried (magnésium sulfate) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 120 g RediSep™ silica cartridge) eluting with a gradient of 0 - 35% ethyl acetate in iso20 heptane afforded the desired regioisomeric products:
\-(6-Cldoro-4-ethyl-5-methylpyridazin-3-yl)-L3-benzothiazol-2-amine as a yellow solid (330 mg, 1.08 mmol, 13%) [Préparation 9fb].
LC/MS (C14H13CIN4S) 305 [M+H]+; RT 1.31 (LCMS-V-B1)
Ή NMR (400 MHz, DMSO-d6) δ 7.82 (br s, 1H), 7.38 (br s + t, 2H), 7.21 (t, 1H), 2.91 (q, J 25 = 7.4 Hz, 2H), 2.38 (s, 3H), 1.12 (t, J = 7.5 Hz, 3H).
N-(6-Chloro-5-ethyl-4-methylpyridazin-3-yl)-l,3-benzothiazol-2-amine as a yellow solid (750 mg, 2.46 mmol, 30%) [Préparation 9fa].
LC/MS (C14H13CIN4S) 305 [M+H]+; RT 1.30 (LCMS-V-B1)
Ή NMR (400 MHz, DMSO-d6) δ 7.86 (s, 1H), 7.49 (br s, 1H), 7.40 (t, 1H), 7.22 (t, 1H), 30 2.79 (q, J = 7.5 Hz, 2H), 2.41 (s, 3H), 1.14 (t, J = 7.5 Hz, 3H).
153
Préparation 9ga and 9gb: /V-{5-[(Benzyloxy)methyl]-6-chloro-4-methylpyridazin-3-yl}l,3-benzothiazol-2-amine and A-{4-[(Benzyloxy)methyl]-6-chloro-5-methylpyridazin-3yl}-l,3-benzothiazol-2-amine
To a solution of the product from Préparation 8g (1.16 g, 4.4 mmol, 1 eq) and 2-bromo-l,3benzothiazole (1.22 g, 5.72 mmol, 1.3 eq) in 1,4-dioxane (30 mL) was added sodium hydride (60% dispersion; 352 mg, 8.8 mmol, 2 eq) portionwise and the reaction was heated at reflux for 2 h. The mixture was allowed to cool to ambient température, quenched with acetic acid and neutralised 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 (magnésium sulfate) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 120 g RediSep™ silica cartridge) eluting with a gradient of 0 - 30% ethyl acetate in isoheptane afforded the desired regioisomeric products:
A-{4-[(Benzyloxy)methyl]-6-chloro-5-methylpyridazin-3-yl}-l,3-benzothiazol-2-amine as a yellow solid (250 mg, 0.63 mmol, 14%) [Préparation 9gb].
LC/MS (C20H17CIN4OS) 398 [M+H]+; RT 1.43 (LCMS-V-B1)
Ή NMR (400 MHz, DMSO-d6) 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).
A-{5-[(Benzyloxy)methyl]-6-chloro-4-methylpyridazin-3-yl}-l,3-benzothiazol-2-amine as a yellow solid (253 mg, 0.64 mmol, 14%) [Préparation 9ga].
LC/MS (C20H17CIN4OS) 398 [M+H]+; RT 1.39 (LCMS-V-B1)
Ή NMR (400 MHz, DMSO-d6) 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).
Préparation 9ha: A-(6-Chloro-5-cyclopropyl-4-methylpyridazin-3-yl)-l,3benzothiazol-2-amine
To a solution of the product from Préparation 8ha (2.44 g, 13.3 mmol, 1 eq) and 2-bromo-l,3benzothiazole (3.7 g, 17.3 mmol, 1.3 eq) in 1,4-dioxane (50 mL) was added sodium hydride (60% dispersion; 1.06 g, 26.6 mmol, 2 eq) portionwise and the mixture was heated at reflux for 1 h. The reaction was allowed to cool to ambient température, then quenched with acetic acid and neutralised by the addition of sodium bicarbonate. The résultant precipitate was collected by filtration and dried under vacuum to afford the desired product (0.98 g, 3.09 mmol, 23%).
LC/MS (C15H13CIN4S) 317 [M+H]+; RT 1.30 (LCMS-V-B1)
154 © Ή NMR (400 MHz, DMSO-d6) 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).
Préparation 9hb: A-(6-Chloro-4-cyclopropyl-5-methylpyridazin-3-yl)-l,3- benzothiazoI-2-amine
To a solution of 2-bromo-l,3-benzothiazole (2.8 g, 13.1 mmol, 1.2 eq) and the product from Préparation 8hb (2 g, 10.9 mmol, 1 eq) in 1,4-dioxane (50 mL) was added sodium hydride (60% dispersion; 0.52 g, 21.8 mmol, 2 eq) portionwise and the reaction was heated at reflux for 1 h. The mixture was allowed to cool to ambient température and concentrated in vacuo. The residue 10 was taken-up in ethyl acetate, filtered, and the solid residue extracted with ethyl acetate. The combined organic extracts were washed with brine, dried (magnésium sulfate) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 40 g RediSep™ silica cartridge) eluting with a gradient of 0 — 40% ethyl acetate in Ao-heptane afforded the desired product as a yellow solid (219 mg, 0.69 mmol, 6%).
LC/MS (C15H13CIN4S) 317 [M+H]+; RT 1.28 (LCMS-V-B1)
Ή NMR (400 MHz, DMSO-d6) 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).
Préparation 9ia: A-[6-Chloro-5-(methoxymethyl)-4-methylpyridazin-3-yI]-1,320 benzothiazol-2-amine
To a solution of the product from Préparation 8ja (323 mg, 1 eq) and 2-bromo-l,3benzothiazole (442 mg, 2.07 mmol, 1.2 eq) in 1,4-dioxane (10 mL) was added sodium hydride (60% dispersion; 138 mg, 3.44 mmol, 2 eq) portionwise and the mixture was heated at reflux for 1.5 h. The mixture was cooled to ambient température, neutralised with acetic acid and 25 concentrated in vacuo. The crude material was partitioned between dichloromethane and water, and the organic phase was dried (magnésium sulfate) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 40 g RediSep™ silica cartridge) eluting with a gradient of 0 - 50% ethyl acetate in Ao-heptane afforded the desired product as a bright yellow solid (85 mg, 0.26 mmol, 15%).
LC/MS (C14H13CIN4OS) 321 [M+H]+; RT 1.19 (LCMS-V-B1)
155 φ Ή NMR (400 MHz, DMSO-d6) δ 8.18 - 6.98 (m, 5H), 4.58 (s, 2H), 3.35 (s, 3H), 2.46 (s,
3H).
Préparation 9jb: /V-[6-Chloro-4-(methoxymethyl)-5-methylpyridazin-3-yl]-l,3benzothiazol-2-amine
To a solution of the product from Préparation 8jb (140 mg, 0.81 mmol, 1 eq) and 2-bromo-l,3benzothiazole (224 mg, 1.05 mmol, 1.3 eq) in 1,4-dioxane (7 mL) was added sodium hydride (60% dispersion; 58.1 mg, 2.42 mmol, 3 eq) portionwise and the mixture was heated at reflux for 30 min. The reaction 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 10 (magnésium sulfate) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 24 g RediSep™ silica cartridge) eluting with a gradient of 0 - 100% ethyl acetate in Ao-heptane afforded the desired product as a yellow solid (169 mg, 0.53 mmol, 58%).
LC/MS (C14H13CIN4OS) 321 [M+H]+; RT 1.23 (LCMS-V-B1)
Ή NMR (400 MHz, DMSO-d6) 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).
Préparation 9ka: /V-[6-Chloro-5-(ethoxymethyl)-4-methylpyridazin-3-yl]-l,3benzothiazol-2-amine
To a solution of the product from Préparation 8ka (136 mg, 0.67 mmol, 1 eq) and 2-bromo-l,320 benzothiazole (173 mg, 0.81 mmol, 1.2 eq) in 1,4-dioxane (10 mL) was added sodium hydride (60% dispersion; 53.9 mg, 1.35 mmol, 2 eq) portionwise and the mixture was heated at reflux for 1 h. The reaction was allowed to cool to ambient température 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 (magnésium sulfate) and concentrated in 25 vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 24 g
RediSep™ silica cartridge) eluting with a gradient of 0 - 40% ethyl acetate in Ao-heptane afforded the desired product as a yellow solid (113 mg, 0.34 mmol, 50%).
LC/MS (C15H15CIN4OS) 335 [M+H]+; RT 1.28 (LCMS-V-B1)
Ή NMR (400 MHz, DMSO-d6) δ 7.86 (s, 1H), 7.46 (br s, 1H), 7.41 (t, 1H), 7.23 (t, J = 7.7 30 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).
156
Préparation 9ma: 7V-{5-[3-(Benzyloxy)propyl]-6-chloro-4-methylpyritllazin-3-yl}-l,3benzothiazol-2-amme
To a solution of the product from Préparation 8ma (1.3 g, 4.46 mmol, 1 eq) and 2-bromo-l,3benzothiazole (1.14 g, 5.35 mmol, 1.2 eq) in 1,4-dioxane (50 mL) was added sodium hydride (60% dispersion; 356 mg, 8.91 mmol, 2 eq) portionwise and the mixture was heated at reflux for 1 h. The reaction was allowed to cool to ambient température then quenched by the addition of water and partitioned between ethyl acetate (100 mL) and sodium bicarbonate (50 mL). The organic phase was washed with brine (50 mL), dried (magnésium sulfate), and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 80 g RediSep™ silica cartridge) eluting with a gradient of 0 - 55% ethyl acetate in Ao-heptane afforded the desired product as a cream solid (1.59 g, 3.74 mmol, 84%).
LC/MS (C22H2iC1N4OS) 425 [M+H]+; RT 1.25 (LCMS-V-B1)
Hl NMR (400 MHz, DMSO-d6) 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).
Préparation 10a: l-(l-Adamantylmethyl)-5-methyl-4-(4,4,5,5-tetramethyI-l,3,2dioxaborolan-2-yl)pyrazole
Step A: 1 -(1-adamantylmethyl) -4-iodo-pyrazole
The mixture of 35.9 g of 1-adamantylmethanol (216 mmol), 73.48 g of triphenylphosphine (280 mmol, 1.3 eq.), 54.25 g of 4-iodo-LH-pyrazole (280 mmol, 1.3 eq.) and 64.4 g of teri-butyl N(teri-butoxycarbonyliminomethylene)carbamate (266 mmol. 1.3 eq.) in 1078 mL of THF was stirred at rt for 48 h. After the addition of extra 10.94 g of 4-iodo-LH-pyrazole (56 mmol, 0.26 eq.), 12.81 g of ieri-butyl V-(/m-butoxycarbonyliminomethylenc)carbamate (53 mmol, 0.26 eq.) and 14.69 g of triphenylphosphine (56 mmol, 0.26 eq.), the reaction was stirred at rt for 24 h then concentrated, purified via flash column chromatography using DCM as eluent, triturated in cold MeOH, and filtered off to give 53.6 g (73%) of the desired product.
Step B: 1 -(1 -adamantylmethyl)-4-iodo-5-methyl-pyrazole
To 9.8 mL of diisopropylamine (69.5 mmol, 1.-1 eq.) in 180 mL of THF was added dropwise 33.4 mL of a 2.5 M solution of butyl lithium (84 mmol, 1.3 eq.) at -78 °C 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 methyliodide (73.3 mmol, 1.14 eq.), and stirred at -78 °C for 18 h. After quenching with cc. NH4C1, the reaction was
157 extracted with EtOAc and the combined organic phases were washed with brine, dried, concentrated, tnturated in MeOH, and filtered off to give 21 g (92%) of the desired product.
Ή NMR (400 MHz, CDC13) 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 -(l-adamantylmethyl)-5-methyl-4-(4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2yl)pyrazole
To 21 g of the product from Step B (58.95 mmol, 1 eq.) in 300 mL of THF was added 28.3 mL of a 2.5 M solution of butyllithium (70.8 mmol, 1.2 eq) at -78 °C and the mixture was stirred at -78 °C for 0.5 h, treated with 16.4 g of 2-isopropoxy-4,4,5,5-tetramethyl-l,3,2-dioxaborolane (88.1 mmol, 1.5 eq.) (addition in portions over 40 min), and kept at -78 °C for 24 h. After quenching with cc. NH4CI at rt, the reaction was extracted with EtOAc and the combined organic phases were washed with brine, dried, concentrated, triturated in MeOH, and filtered off to give 19.7 g (94%) of the desired product.
Ή NMR (500 MHz, DMSO-î/6) 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); 13C NMR (500 MHz,, DMS0-d6) δ ppm 146.9, 144.1, 104.6, 59.7, 40.6, 36.8, 35.4, 28.1, 25.1, 12.1; HRMS-ESI (m/z): [M+H]+ calcd for C21H34BN2O2: 357.2713, found 357.2704.
Préparation 10b: l-{[l-(3-Methoxypropyl)cyclooctyl]methyl}-5-methyl-4-(4,4,5,5tetramethyl-l,3,2-dioxaborolan-2-yl)-LH-pyrazole
Step A: methyl l-(3-methoxypropyl)cyclooctanecarboxylate
To 4.74 g (1.14 eq.) of diisopropylamine in 90 mL of tetrahydrofuran was added 18.8 mL (1.14 eq.) of a 2.5 Μ solution of butyl lithium at -78 °C and after 0.5 h at -78 °C, 7.0 g (41.1 mmol) of methyl cyclooctanecarboxylate in 40 mL of tetrahydrofuran was added over 1 h. After 1 h at -78 °C, 7.2 g (1.14 eq.) of l-bromo-3-methoxy-propane was added and the mixture was stirred for 18 h. After quenching the reaction with the addition of saturated NH4CI solution, the mixture was extracted with EtOAc and the organic phases were dried over MgSÛ4 and concentrated to give 8.0 g (80%) of the desired product.
Ή NMR (400 MHz, CDCI3) 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: [l-(3-methoxypropyl)cyclooctyl]methanol
158
P Το 9.0 g (37.13 mmol) of the product from Step A in 93 mL of diethyl ether was added 1.76 g (1.25 eq.) of lithium aluminum hydride portion wise at 0 °C. After stirring at rt for 2 h, the reaction was quenched by the addition of icy water and EtOAc and a 10% solution of NaOH were added. The mixture was extracted with EtOAc, dried, and concentrated to give 7.4 g (93%) 5 of the desired product.
NMR (400 MHz, CDC13) δ ppm 3.37 (t, 2H), 3.34 (s, 3H), 3.30 (s, 2H), 1.61-1.23 (m, 18H).
Step C: 4-iodo-l-[[l-(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.) of 4-iodo-lH-pyrazole in 33 mL of tetrahydrofuran was added 2.22 g (1.3 eq.) of triphenylphosphine and 1.95 g (1.3 eq.) 10 of diimbutyl azodicarboxylate and the mixture was stirred at rt for 67 h. To the mixture was added 278 mg of 4-iodo-lH-pyrazole, 444 mg of triphenylphosphine, and 390 mg of ditertbutyl azodicarboxylate and was stirred at rt for 24 h. After the addition of reagents and stirring at rt for 24 h was repeated (115 h stirring in total), the mixture was concentrated and purified via flash column chromatography (silica gel) using heptane and EtOAc as eluents to give 1.24 g 15 (49%) of the desired product.
Ή NMR (400 MHz, CDCI3) δ ppm 7.47 (s, 1H), 7.42 (s, 1H), 3.93 (s, 2H), 3.37 (t, 2H), 3.36 (s, 3H), 1.68-1.18 (m, 18H).
Step D: 4-iodo-l-[[l-(3-methoxypropyl)cyclooctyl]methyl]-5-methyl-pyrazole
To 1.2 g (3.07 mmol) of the product from Step C in 5 mL of tetrahydrofuran was added 3.7 mL 20 (1.2 eq.) of a 1 M solution of LDA at -78 °C. After 0.6 h at -78 °C, 0.5 mL (1.14 eq.) of methyl iodide was added dropwise to the mixture and it was let to warm up to rt over 20 h. Reaction was quenched with a saturated solution of NH4CI and extracted with EtOAc. The combined organic phases were dried, concentrated, and purified via flash column chromatography (silica gel) using heptane and EtOAc as eluents to give 0.79 g (64%) of the desired product.
Ή NMR (400 MHz, CDCI3) δ ppm 7.43 (s, 1H), 3.85 (s, 2H), 3.38 (t, 2H), 3.35 (s, 3H), 2.29 (s, 3H), 1.69-1.24 (m, 18H).
159
Step E: l-[[l-(3-methoxypropyl)cyclooctyl]methyl]-5-methyl~4-(4,4,5,5-tetramethyl-l,3,2dioxaborolan-2-yl)pyrazole
To the solution of 0.81 g (2 mmol) of the product from Step D in 15 mL of tetrahydrofuran was added 0.96 mL (1.2 eq.) of a 2.5 M solution of butyl lithium dropwise at -78 °C. After 0.5 h, 0.5 mL (1.2 eq.) of 2-isopropoxy-4,4,5,5-tetramethyl-l,3,2-dioxaborolane was added over 20 min and the mixture was kept at -78 °C for 6 h and at rt for 6 h. After quenching the reaction with saturated solution of NH4CI and extracting with EtOAc, the combined organic phases were washed with brine, dried, and purified via flash column chromatography (silica gel) using heptane and EtOAc as eluents to give 0.33 g (34%) of the desired product.
'H NMR (500 MHz, dmso-d6) 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). 13C NMR (500 MHz, dmso-d6) 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-ESI (m/z): [M+H]+ calcd for C23H42N2O3B: 405.3289, found 404.3334.
Préparation 10c: l-{[l-(3-Methoxypropyl)cyclohexyI]methyl}-5-metbyl-4-(4,4,5,5tetramethyl-l,3,2-dioxaborolan-2-yl)-lH-pyrazole
Step A: methyl l-(3-methoxypropyl)cyclohexanecarboxylate
To 6.84g (1.09 eq.) of diisopropylamine in 130 mL of tetrahydrofuran was added 27 mL (1.09 eq.) of a 2.5 M solution of butyl lithium at -78 °C and after 0.5 h at -78 °C, 8.8 g of methyl cyclohexanecarboxylate in 50 mL of tetrahydrofuran was added over 1 h. After 1 h at -78 °C, 10.7 g (1.13 eq.) of l-bromo-3-methoxy-propane was added and the mixture was stirred for 18 h. After quenching the reaction with the addition of saturated NH4CI solution, the mixture was extracted with EtOAc and the organic phases were dried over MgSÜ4 and concentrated to give 12 g (92%) of the desired product.
Ή NMR (400 MHz, CDCI3) δ ppm 3.67 (s, 3H), 3.35 (d, 1H), 3.32 (d, 1H), 3.31 (s, 3H), 2.112.03 (m, 2H), 1.60-1.16 (m, 12H).
Step B: [l-(3-methoxypropyl)cyclohexyl]methanol
To 12 g (56.41 mmol) of the product from Step A in 140 mL of diethyl ether was added 2.68 g (1.25 eq.) of lithium aluminum hydride portion wise at 0 °C. After stirring at rt for 2 h, the reaction was quenched by the addition of icy water and EtOAc and a 10% solution of NaOH
160 were added. The mixture was extracted with EtOAc, dried, and concentrated to give 9.37 g (89%) of the desired product.
Ή NMR (400 MHz, CDC13) δ ppm 3.41 (s, 2H), 3.38 (t, 2H), 3.35 (s, 3H), 1.56-1.27 (m, 14H).
Step C: 4-iodo-l-[[1 -(3-methoxypropyl)cyclohexyl]methyl]pyrazole
To 1.21g (6.5 mmol) of the product from Step B and 2.58 g (2.05 eq.) of 4-iodo-l/f-pyrazole in 33 mL of tetrahydrofuran was added 3.5 g (2.05 eq.) of triphenylphosphine and 3.07 g (2.05 eq.) of diiertbutyl azodicarboxylate and the mixture was stirred at rt for 2 h. To the mixture was added 140 mg of 4-iodo-lH-pyrazole, 230 mg of triphenylphosphine, and 200 mg of diiertbutyl azodicarboxylate and was stirred at rt for 24 h. After the addition of reagents and stirring at rt for 24 h was repeated twice (96 h stirring in total), the mixture was concentrated and purified via flash column chromatography (silica gel) using heptane and EtOAc as eluents to give 1.4 g (59.5%) of the desired product.
Ή NMR (400 MHz, CDCI3) δ ppm 7.47 (s, 1H), 7.41 (s, 1H), 4.00 (s, 2H), 3.36 (t, 2H), 3.35 (s, 3H), 1.62-1.21 (m, 14H).
Step D: 4-iodo-l-[[l-(3-methoxypropyl)cyclohexyl]methyl]-5-methyl-pyrazole
To 3.7 g (10.21 mmol) ofthe product from Step C in 15 mL of tetrahydrofuran was added 12.3 mL (1.2 eq.) of a 1 M solution of LDA in tetrahydrofuran at -78 °C. After 0.6 h at -78 °C, 0.73 mL (1.14 eq.) of methyl iodide was added dropwise to the mixture and it was let to warm up to rt over 20 h. Reaction was quenched with a saturated solution of NH4CI and extracted with EtOAc. The combined organic phases were dried, concentrated, and purified via flash column chromatography (silica gel) using heptane and EtOAc as eluents to give 2.85 g (74%) of the desired product.
Ή NMR (400 MHz, CDCI3) δ ppm 7.44 (s, 1H), 3.92 (s, 2H), 3.38 (t, 2H), 3.35 (s, 3H), 2.29 (s, 3H), 1.58-1.13 (m, 14H).
Step E: l-[[l-(3-methoxypropyl)cyclohexyl]methyl]-5-rnethyl-4-(4,4,5,5-tetramethyl-l,3,2dioxaborolan -2-yl) pyrazole
To the solution of 5.0 g (13.3 mmol) of the product from Step D in 71 mL of tetrahydrofuran was added 6.38 mL (1.2 eq.) of a 2.5 M solution of butyl lithium dropwise at -78 °C. After 0.5 h, 4.1 mL (1.5 eq.) of 2-isopropoxy-4,4,5,5-tetramethyl-l,3,2-dioxaborolane was added over 40 min and the mixture was kept at -78 °C for 6 h and at rt for 6 h. After quenching the reaction with saturated solution of NH4CI and extracting with EtOAc, the combined organic phases were
161
B washed with brine, dried, and purified via flash column chromatography (silica gel) using heptane and EtOAc as eluents to give 2.3 g (46%) of the desired product.
Ή NMR (500 MHz, dmso-d6) 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). 13C NMR (500 MHz, 5 dmso-d6) 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-ESI (m/z): [M+H]+ calcd for C21H38N2O3B: 376.2897, found 376.3019.
Préparation lia: Ethyl 5-bromo-2-[(4,5-dimethyI-6-{[(2Z)-3-{[2(trimethylsilyl)ethoxy]methyl}-2,3-dihydro-l,3-benzothiazol-2-ylidene]amino}pyridazin10 3-yl)amino]-l,3-thiazole-4-carboxylate
Step A: (2Z)-N-(6-chloro-4,5-dimethylpyridazin-3-yl)-3-{[2-(trimethylsilyl)ethoxy]methyl}-
2,3- dihydro-1,3-benzothiazol-2-imine
A,A-Diisopropylethylamine (3.6 mL, 20.6 mmol, 2 eq) was added to a suspension of the product from Préparation 9a (3 g, 10.3 mmol, 1 eq) in dichloromethane (75 mL) at 0 °C under 15 a nitrogen atmosphère. [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 température for 3 h. Dichloromethane (75 mL) was added and the mixture washed with water (2 x 75 mL) and brine (75 mL). The solution was dried (magnésium sulfate) and concentrated in vacuo. Purification by column chromatography, isolute flash silica (20 g) 20 eluting with dichloromethane afforded the desired product as a pale brown solid (3.95 g, 8.44 mmol, 82%) that was used directly in the next step without further characterisation.
Step B: ethyl 2-[(4,5-dimethyl-6-{[(2Z)-3-{[2-(trimethylsilyl)ethoxy]methyl}-2,3-dihydro-l,3benzothiazol-2-ylidene]amino}pyridazin-3-yl)amino]-l,3-thiazole-4-carboxylate
The product from Step A (500 mg, 1.19 mmol, 1 eq) and ethyl 2-amino-l,3-thiazole-425 carboxylate (245.4 mg, 1.43 mmol, 1.2 eq) were added to a solution of césium carbonate (1.16 g, 3.56 mmol, 3 eq), tris(dîbenzylideneacetone)dipalladium(0) (54.4 mg, 0.06 mmol, 0.05 eq) and Xantphos (68.7 mg, 0.12 mmol, 0.1 eq) in alpha,alpha,alphaArrAuototoluene (10 mL) under a nitrogen atmosphère. The mixture was sparged with nitrogen (10 min) then heated at 170 °C for 1 h under microwave irradiation. The solution was allowed to cool to ambient 30 température and concentrated in vacuo. The mixture was partitioned between ethyl acetate and water and the organic phase was washed with brine, dried (magnésium sulfate) and solvent removed in vacuo. Purification by automated flash column chromatography (CombiFlash Rf,
162 g RediSep™ silica cartridge) eluting with a gradient of 0 - 50% ethyl acetate in Ao-heptane afforded the desired product as an orange foam (301 mg, 0.54 mmol, 46%).
LC/MS (C25H32N6O3SiS2) 557 [M+H]+; RT 1.49 (LCMS-V-B1)
Ή NMR (400 MHz, DMSO-d6) 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-{[(2Z)-3-{[2-(trimethylsilyl)ethoxy]methyl}-2,3dihydro-l,3-benzothiazol-2-ylidene]amino}pyridazin-3-yl)amino]-l,3-thiazole-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 was stirred at ambient température for 1.5 h then the solvent was removed in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 24 g RediSep™ silica cartridge) eluting with a gradient of 0 - 60% ethyl acetate in Ao-heptane afforded the desired product as a brown gum (198 mg, 0.31 mmol, 58%).
LC/MS (C25H3iBrN6O3SiS2) 637 [M+H]+; RT 1.57 (LCMS-V-B1) ‘H NMR (400 MHz, DMSO-d6) 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).
Préparation 11b: Ethyl 5-bromo-2-[methyl(5-methyl-6-{[(2Z)-3-{[2(trimethyIsilyl)ethoxy]methyl}-2,3-dihydro-l,3-benzothiazoI-2-ylidene]amino}pyridazin3-yl)amino]-l,3-thiazole-4-carboxylate
Step A: ethyl2-[(6-chloro-5-methylpyridazin-3-yl)(methyl)amino]-l,3-thiazole-4carboxylate
A solution of 3,6-dichloro-4-methyl-pyridazine (662 mg, 4.06 mmol, 1.02 eq) and ethyl 2(methylamino)-l,3-thiazole-4-carboxylate (742 mg, 3.98 mmol, 1 eq) in tetrahydrofuran (20 mL) was cooled in an ice bath then sodium hydride (60% in minerai oil; 188 mg, 4.7 mmol, 1.18 eq) was added slowly. The mixture was maintained in the ice bath for 1 h then stirred at ambient température for 3 h. Saturated aqueous ammonium chloride solution was added and the solution was extracted with dichloromethane (x2). The combined organic extracts were washed with water (x2) followed by brine then dried (magnésium sulfate) and the solvent
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removed in vacuo. The residue was triturated in a mix of dichloromethane and methanol and the solids collected by filtration and dried under vacuum to afford the desired product as a cream solid (324 mg, 1.04 mmol, 26%).
LC/MS (C12H13CIN4O2S) 313 [M+H]+; RT 1.15 (LCMS-V-B1)
Ή NMR (400 MHz, DMSO-d6) 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-[(l,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)l,3-thiazole-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 césium carbonate (1.46 g, 4.47 mmol, 3 eq), 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-tüüuorotoluene (10 mL) under a nitrogen atmosphère. The mixture was sparged with nitrogen (10 min) then heated at 170 °C for 1 h under microwave irradiation. The solution was allowed to cool to ambient température, filtered, eluting with dichloromethane, and the filtrate concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 24 g RediSep™ silica cartridge) eluting with a gradient of 0 - 5% methanol in dichloromethane afforded the desired product as a yellow solid (133 mg, 0.31 mmol, 21%).
LC/MS (C19H18N6O2S2) 427 [M+H]+; RT 1.30 (LCMS-V-B1)
Hl NMR (400 MHz, DMSO-d6) δ 11.10 (s, 1 H), 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-{[(2Z)-3-{[2-(trimethylsilyl)ethoxy]methyl}-2,3-dihydrol,3-benzothiazol-2-ylidene]amino}pyridazin-3-yl)amino]-l,3-thiazole-4-carboxylate
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 Α,Α-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 slowly warm to ambient température and was stirred overnight. The reaction was diluted with dichloromethane, washed successively with water (x2) and brine, dried (magnésium sulfate), and the solvent removed in vacuo. Purification by automated flash
164 column chromatography (CombiFlash Rf, 12 g RediSep™ silica cartridge) eluting with a gradient of 0 - 50% ethyl acetate in Ao-heptane afforded the desired product as a yellow glass (56.3 mg, 0.1 mmol, 32%).
LC/MS (C25H32N6O3SiS2) 557 [M+H]+; RT 1.64 (LCMS-V-B1)
Ή NMR (400 MHz, DMSO-d6) δ 7.97 (s, 1H), 7.85 (dd, J = 7.6, 1.0 Hz, 1H), 7.71 (d, J = 1.1 Hz, 1H), 7.50 - 7.40 (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-{[(2Z)-3-{[2-(trimethylsilyl)ethoxy]methyl}-2,3dihydro-l,3-benzothiazol-2-ylidene]amino}pyridazin-3-yl)amino]-l,3-thiazole-4-carboxylate
7V-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 was stirred at ambient température for 1.5 h then the solvent removed in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 24 g RediSep™ silica cartridge) eluting with a gradient of 0 - 60% ethyl acetate in Ao-heptane afforded the desired product as a white solid (54.7 mg, 0.09 mmol, 85%)
LC/MS (C25H3iBrN6O3SiS2) 635 [M+H]+; RT 1.73 (LCMS-V-B1)
Ή NMR (400 MHz, DMSO-d6) 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).
Préparation 11c: Ethyl 5-bromo-2-[(3-methoxypropyl)(5-methyl-6-{[(2Z)-3-{[2(trimethylsilyI)ethoxy]methyl}-2,3-dihydro-l,3-benzothiazol-2-ylidene]ainino}pyridazin3-yl)amino]-l,3-thiazoIe-4-carboxylate
Step A: ethyl2-[(3-methoxypropyl)amino]-l,3-thiazole-4-carboxylate
Ammonia (2M in isopropanol; 60 mL, 0.12 mol, 3.45 eq) was added to a solution of 3methoxypropylisothiocyanate (4.5 g, 34.3 mmol, 1 eq) in isopropanol (15 mL) at 0 °C and the mixture was stirred for 1 h at 0 °C then for 18 h at ambient température. The suspension was concentrated in vacuo to afford a pale yellow gum. 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 a nitrogen atmosphère. Triethylamine (10 mL, 68.6 mmol, 2 eq) was added and the mixture was stirred at
165 ambient température for 18 h. The reaction mixture was partitioned between ethyl acetate (300 mL) and water, and the organic phase was successively washed with water (150 mL) and brine (100 mL), dried (magnésium sulfate) and concentrated in vacuo. Purification by flash column chromatography (100 g silica) eluting with 99:1 dichloromethane / methanol gave a dark yellow oil that was triturated heptane (30 mL), filtered, washed with heptane (20 mL) and dried under vacuum afford the desired product as a pale brown powder (5.75 g, 23.5 mmol, 69%).
LC/MS (C10H16N2O3S) 245 [M+H]+; RT 0.99 (LCMS-V-B1) *H NMR (400 MHz, CDCI3) 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), 3.54 - 3.42 (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]-l,3-thiazole-4carboxylate
Sodium hydride (60% dispersion; 350 mg, 8.75 mmol, 1.22 eq) 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 under a nitrogen atmosphère and the mixture was stirred for 1 h at 0 °C then heated at 60 °C for 6 h. The reaction mixture was allowed to cool to ambient température then partitioned between ethyl acetate (300 mL) and saturated aqueous ammonium chloride (150 mL). The organic phase was successively washed with water (2 x 150 mL) and brine (100 mL), dried (magnésium sulfate), and concentrated in vacuo. Purification by flash column chromatography (50 g silica) eluting with 39:1 dichloromethane / methanol gave a brown oil that was triturated with diethyl ether (10 mL), filtered, washed with diethyl ether (10 mL) and dried under vacuum afford the desired product as a pale brown powder (1.3 g, 3.51 mmol, 49%).
LC/MS (C15H19CIN4O3S) 371 [M+H]+; RT 1.28 (LCMS-V-B1) XH NMR (400 MHz, CDCI3) 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).
Step C: ethyl 2-({6-[(l,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(3methoxypropyl)amino)-l,3-thiazole-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,
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0.18 mmol, 0.05 eq) and Xantphos (203 mg, 0.35 mmol, 0.1 eq) in 1,4-dioxane (20 mL) under a nitrogen atmosphère and the mixture was stirred. AW-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 h. The solution was allowed to cool to ambient température and concentrated in vacuo. Purification by flash column chromatography (50 g silica) eluting with 99:1 dichloromethane / methanol gave a red gum that was triturated with methanol (15 mL), filtered, washed with methanol (10 mL) and dried under vacuum to afford the desired product as a yellow solid (1.35 g, 2.79 mmol, 80%).
LC/MS (C22H24N6O3S2) 485 [M+H]+; RT 1.41 (LCMS-V-B1) XH NMR (400 MHz, DMSO-d6) 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-{[(2Z)-3-{[2-(trimethylsilyl)ethoxy]methyl}2,3-dihydro-l,3-benzothiazol-2-ylidene]amino}pyridazin-3-yl)amino]-l,3-thiazole-4carboxylate
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 Α,Α-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 was allowed to warm to ambient température and stirred for 5 h. The mixture was diluted with dichloromethane then washed with water followed by brine, dried (magnésium sulfate), and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 24 g RediSep™ silica cartridge) eluting with a gradient of 0 - 50% ethyl acetate in Ao-heptane afforded the desired product as a cream foam (0.83 g, 1.35 mmol, 76%).
LC/MS (C28H3sN6O4SiS2) 615 [M+H]+; RT 1.51 (LCMS-V-B1)
Ή NMR (400 MHz, DMSO-d6) 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).
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Step E: ethyl 5-bromo-2-[(3-methoxypropyl) (5-methyl-6-{[(2Z)-3-{[2(trimethylsilyl)ethoxy]methyl}-2,3-dihydro-l,3-benzothiazol-2-ylidene]amino}pyridazin-3yl)amino]-l,3-thiazole-4-carboxylate
Æ-Bromosuccinimide (63.7 mg, 0.36 mmol, 1.1 eq) was added to a stirred solution of the product of Step A (200 mg, 0.33 mmol, 1 eq) in dichloromethane (10 mL) and the mixture was stirred for 4 h. The solvent was removed in vacuo and purification by automated flash column chromatography (CombiFlash Rf, 12 g RediSep™ silica cartridge) eluting with a gradient of 0 - 60% ethyl acetate in Ao-heptane afforded the desired product as a cream foam (193 mg, 0.28 mmol, 85%).
LC/MS (C28H37BrN6O4SiS2) 695 [M+H]+; RT 1.60 (LCMS-V-B1) *H NMR (400 MHz, DMSO-d6) 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).
Préparation lld: Ethyl 5-bromo-2-[(5-cyclopropyl-6-{[(2Z)-3-{[2(trimethylsilyl)ethoxy]methyl}-2,3-dihydro-l,3-benzothiazol-2-yIidene]amino}pyridazin3-yl)(methyl)amino]-l,3-thiazole-4-carboxylate
Step A: ethyl2-[(6-chloro-5-cyclopropylpyridazin-3-yl)(methyl)amino]-l,3-thiazole-4carboxylate
To a solution of the product from Préparation 71 (495 mg, 2.62 mmol, 1 eq) and ethyl 2(methylamino)-l,3-thiazole-4-carboxylate (488 mg, 2.62 mmol, 1 eq) in tetrahydrofuran (25 mL) at 0 °C was added slowly sodium hydride (60% in minerai oil; 124 mg, 3.09 mmol, 1.18 eq) under a nitrogen atmosphère. The mixture was stirred for 1 h at 0 °C, then allowed to warm to ambient température and stirred for 18 h. Saturated aqueous ammonium chloride solution (75 mL) was added and the mixture extracted with dichloromethane (250 mL). The organics were successively washed with water (2 x 75 mL) and brine (75 mL), dried (magnésium sulfate) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 40 g RediSep™ silica cartridge) eluting with a gradient of 0 - 60% ethyl acetate in Ao-heptane afforded the desired product as a white crystalline solid (398 mg, 1.17 mmol, 45%).
LC/MS (C14H15CIN4O2S) 339 [M+H]+; RT 1.25 (LCMS-V-B1)
168
Ή NMR (400 MHz, DMSO-d6) 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).
Step B: ethyl 2-({6-[(l,3-benzothiazol-2-yl)amino]-5-cyclopropylpyridazin-3yl}(methyl)amino)-l,3-thiazole-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 to a solution of Xantphos (68 mg, 0.12 mmol, 0.1 eq) and tris(dibenzylideneacetone)dîpalladium(0) (53.8 mg, 0.06 mmol, 0.05 eq) in 1,4-dioxane (20 mL) under a nitrogen atmosphère. A,A-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 température and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 40 g RediSep™ silica cartridge) eluting with a gradient of 0 - 90% ethyl acetate in Ao-heptane afforded the desired product as a yellow solid (249 mg, 0.55 mmol, 47%).
LC/MS (C21H20N6O2S2) 453 [M+H]+; RT 1.38 (LCMS-V-B1)
Ή NMR (400 MHz, DMSO-d6) δ 7.96 (s, 1H), 7.90 (s, 1H), 7.50 (br s, 1H), 7.39 (t, J = 7.6 Hz, 1H), 7.29 - 7.08 (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-{[(2Z)-3-{[2-(trimethylsilyl)ethoxy]methyl}-2,3-dihydrol,3-benzothiazol-2-ylidene]amino}pyridazin-3-yl)(methyl)amino]-l,3-thiazole-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 VA-diisopropylethylamine (0.16 mL, 0.98 mmol, 2 eq) was added and the mixture was stirred for 10 min. 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 température for 7 h. The reaction was partitioned between dichloromethane and water and the organic phase was washed with water followed by brine, dried (magnésium sulfate), and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 12 g RediSep™ silica cartridge) eluting with a gradient of 0 - 50% ethyl acetate in Ao-heptane afforded the desired product as a cream foam (185 mg, 0.32 mmol, 65%).
LC/MS (C27H34N6O3SiS2) 583 [M+H]+; RT 1.45 (LCMS-V-B1)
169
Ή NMR (400 MHz, DMSO-d6) 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-{[(2Z)-3-{[2-(trimethylsilyl)ethoxy]methyl}-2,3dihydro-l,3-benzothiazol-2-ylidene]amino}pyridazin-3-yl)(methyl)amino]-l,3-thiazole-4carboxylate
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 purification by automated flash column chromatography (CombiFlash Rf, 12 g RediSep™ silica cartridge) eluting with a gradient of 0 - 60% ethyl acetate in Ao-heptane afforded the desired product as a cream solid (149 mg, 0.23 mmol, 71%).
LC/MS (C27H33BrN6O3SiS2) 663 [M+H]+; RT 1.52 (LCMS-V-B1)
Ή NMR (400 MHz, DMSO-d6) 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).
Préparation lie: Ethyl 5-iodo-2-[methyl(5-methyl-6-{[(2Z)-3-{[2(trimethylsilyl)ethoxy]methyl}-2,3-dihydro-l,3-benzothiazol-2-ylidene]amino}pyridazin3-yl)amino]-l,3-thiazole-4-carboxylate
Step A: ethyl2-(methylamino)-l,3-thiazole-4-carboxylate
To a suspension of A-methylthiourea (5 g, 55.5 mmol, 1 eq) in éthanol (80 mL), cooled in icewater, was added ethyl bromopyruvate (7.45 mL, 59.4 mmol, 1.07 eq) slowly. After 10 min the mixture was allowed to warm to ambient température and stirred for 1.5 h. Triethylamine (15.4 mL, 111 mmol, 2 eq) was added and the mixture stirred at ambient température for 18 h. The reaction was concentrated in vacuo, the residue partioned between ethyl acetate and water, the aqueous phase was extracted with ethyl acetate (x2), and the combined organic extracts were dried (magnésium sulphate) and concentraed in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 80 g RediSep™ silica cartridge) eluting wi th a gradient of 0 - 100% ethyl acetate in Ao-heptane afforded the desired product as a beige solid (5.26 g, 28.2 mmol, 51%).
170 φ LC/MS (C7H10N2O2S) 187 [Μ+Η]+; RT 0.85 (LCMS-V-B1)
Ή NMR (400 MHz, DMSO-d6) 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]-l,3-thiazole-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 sodium hydride (60% in minerai oil; 1.24 g, 31.1 mmol, 1.1 eq) was added portionwise. The mixture was allowed to gradually warm to ambient température and was stirred for 18 h. 10 Dichloromethane and saturated aqueous ammonium chloride were added forming a precipitate.
The solid was collected by filtration, washed with water then dried under vacuum at 40 °C for 12 h to afford the desired product as a cream solid (4.24 g, 13.6 mmol, 48%).
LC/MS (C12H13CIN4O2S) 313 [M+H]+; RT 1.19 (LCMS-V-B1)
Hl NMR (400 MHz, DMSO-d6) δ 8.04 (s, 1H), 7.87 (d, J = 1.1 Hz, 1H), 4.30 (q, J = 7.1 Hz, 15 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-[(l,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)1,3-thiazole-4-carboxylate
Tris(dîbenzylideneacetone)dipalladium(0) (0.62 g, 0.68 mmol, 0.05 eq) followed by N,Ndiisopropylethylamine (6.74 mL, 40.7 mmol, 3 eq) were added to a sealed flask containing the 20 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) in 1,4-dioxane (250 mL) under a nitrogen atmosphère and the vessel was sealed and heated at 150 °C for 24 h. The reaction was allowed to cool to ambient température and purification by automated flash column chromatography (CombiFlash Rf, 120 g RediSep™ silica cartridge) eluting with a gradient of 0 - 100% ethyl 25 acetate in zso-heptane afforded the desired product as a beige soild (3.05 g, 7.14 mmol, 53%).
LC/MS (C19H18N6O2S2) 427 [M+H]+; RT 1.16 (LCMS-V-B1) XH NMR (400 MHz, DMSO-d6) 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-{[(2Z)-3-{[2-(trimethylsilyl)ethoxy]methyl}-2,3-dihydrol,3-benzothiazol-2-ylidene]amino}pyridazin-3-yl)amino]-l,3-thiazole-4-carboxylate
171
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 A/A-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 température and stir for 6 h. The reaction mixture was washed with water, followed by brine then dried (magnésium sulphate) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 80 g RediSep™ silica cartridge) eluting with a gradient of 0 - 50% ethyl acetate in Ao-heptane afforded the desired product as a white solid (1.38 g, 2.47 mmol, 35%).
LC/MS (C25H32N6O3S1S2) 557 [M+H]+; RT 1.65 (LCMS-V-B1)
Hl NMR (400 MHz, DMSO-d6) 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-{[(2Z)-3-{[2-(trimethylsilyl)ethoxy]methyl}-2,3dihydro-l,3-benzothiazol-2-ylidene]amino}pyridazin-3-yl)amino]-l,3-thiazole-4-carboxylate
AModosuccinimide (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 ambient température for 18 h. The reaction was successively washed with water and brine, dried (magnésium sulphate) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 12 g RediSep™ silica cartridge) eluting with a gradient of 0 - 60% ethyl acetate in Ao-heptane afforded the desired product as a cream solid (180 mg, 0.26 mmol, 73%).
LC/MS (C25H3iIN6O3SiS2) 683 [M+H]+; RT 1.73 (LCMS-V-B1)
Hl NMR (400 MHz, DMSO-d6) 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).
Préparation llf: Ethyl 5-bromo-2-[(6-{[(2Z)-3-{[2-(trimethylsilyl)ethoxy]methyl}2,3-dihydro-l,3-benzothiazol-2-ylidene]amino}pyridazin-3-yl)amino]-l,3-thiazole-4carboxylate
172
Step A: ethyl2-[(6-{[(2Z)-3-{[2-(trimethylsilyl)ethoxy]methyl}-2,3-dihydro-l,3-benzothiazol2-ylidene]amino}pyridazin-3-yl)amino]-l,3-thiazole-4-carboxylate
The product from Préparation 4b (1 g, 2.54 mmol, 1 eq) and ethyl 2-amino-l,3-thiazole-4carboxylate (500 mg, 2.9 mmol, 1.14 eq) were added to a solution of 5 tris(dibenzylideneacetone)dipalladîum(0) (116.5 mg, 0.13 mmol, 0.05 eq) and Xantphos (147.2 mg, 0.25 mmol, 0.1 eq) in 1,4-dioxane (20 mL) under a nitrogen atmosphère. N,Ndiisopropylethylamine (1.5 mL, 7.63 mmol, 3 eq) was added and the mixture heated at 100 °C for 18 h. The solution was allowed to cool to ambient température and concentrated in vacuo. Purification by column chromatography (50 g silica) eluting with 50% ethyl acetate in 10 heptane gave a dark yellow solid. Trituration with heptane (20 mL) gave a solid that was collected by filtration, washed with heptane (2 x 10 mL) and dried under vacuum to afford the desired product as a yellow solid (995 mg, 1.88 mmol, 74%) [mixture of isorners].
LC/MS (C23H28N6O3SiS2) 529 [M+H]+; RT 1.54 (LCMS-V-B1)
Ή NMR (400 MHz, Chloroform-d) δ 10.83 (s, 1H), 7.90 - 7.29 (m, 6H), 7.26 - 7.13 (m, 15 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-{[(2Z)-3-{[2-(trimethylsilyl)ethoxy]methyl}-2,3~dihydro-l,3benzothiazol-2-ylidene]amino}pyridazin-3-yl)amino]-l,3-thiazole-4-carboxylate
A-Bromosuccinimide (45.2 mg, 0.25 mmol, 1.3 eq) was added to a stirred solution of the 20 product from Step A (100 mg, 0.19 mmol, 1 eq) in dichloromethane (10 mL) and the mixture was stirred at ambient température for 1 h. The solvent was removed in vacuo and purification by automated flash column chromatography (CombiFlash Rf, 12 g RediSep™ silica cartridge) eluting with a gradient of 0 - 80% ethyl acetate in Ao-heptane afforded the desired product as a cream solid (64.1 mg, 0.11 mmol, 56%) [mixture of isomers].
LC/MS (C23H27BrN6O3SiS2) 609 [M+H]+; RT 1.47(LCMS-V-B1)
Ή NMR (400 MHz, DMSO-d6) δ 12.45 (s, 1H), 12.31 (s, 1H), 8.13 - 8.04 (m, 1H), 8.04 7.90 (m, 2H), 7.84 - 7.71 (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).
173
Préparation 12: terAbutyl-diphenyl-[2-[[3,5-dimethyl-7-[[5-methyl-4-(4,4,5,5tetramethyl-l,3,2-dioxaboroIan-2-yl)pyrazol-l-yl]methyl]-ladamanty 1] oxy] ethoxy] silane
Step A: 3-bromo-5,7-dimethyladamantane-l-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, 3,5dimethyladamantane-l-carboxylic acid (25 g, 1 eq) was added and the reaction mixture was stirred at rt for 2 days. After the addition of EtOAc, the reaction mixture was treated carefully with a saturated solution of sodium-thiosulfate at 0 °C and stirred for 15 min. After filtration through a pad of Celite and rinsing with EtOAc, the organic phase was separated, washed with a saturated solution of sodium-thiosulfate and brine, dried, concentrated to give the desired product (34.28 g, 74.6%), which was used without further purification.
Ή NMR (400 MHz, DMSO-d6): 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); 13C NMR (100 MHz, DMSO-d6) δ ppm 176.8, 66.8, 54.0, 48.7, 48.5, 45.7, 43.3, 35.5, 29.4; HRMS-ESI (m/z): [M-H]- calcd for C13H18BrO2: 285.0496; found 285.0498.
Step B: 3-bromo-5,7-dimethyl-l-adamantyl-methanol
To the product from Step A (34.3 g, 119 mmol) in THF (77.6 mL) was added slowly a 1 M solution of BH3-THF in THF (358 mL, 3 eq) and the reaction mixture was stirred for 18 h. After the addition of methanol and stirring for 30 min, purification by column chromatography (silica gel, heptane and MTBE as eluents) afforded the desired product (16.19 g, 49.6%).
JH NMR (400 MHz, DMSO-d6): 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) 13C NMR (100 MHz, DMSO-d6) δ ppm 70.4, 68.9, 54.9, 49.8, 49.3, 43.8, 41.4, 35.7, 29.7; HRMS-ESI (m/z): [M-Br]- calcd for C13H21O: 193.1598 found: 193.1589.
Step C: 1 -[3-bromo-5,7-dimethyl-l -adamantyl]methyl]pyrazole
To the product from Step B (16.19 g, 59.26 mmol) and lH-pyrazole (4.841 g, 1.2 eq) in toluene (178 mL) was added (cyanomethylene)tributylphosphorane (18.64 mL, 1.2 eq) in one portion
174 and the reaction mixture was stirred at 90 °C for 2 h. Purification by column chromatography (silica gel, heptane and MTBE as eluents) afforded the desired product (17.88 g, 93%).
'H NMR (400 MHz, DMSO-dô): 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); 13C NMR (100 MHz, DMSO-d6) 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-ESI (m/z): [M]+ calcd for C16H23BrN2· 322.1045 found: 322.1014.
Step D: 5-methyl-l-[[-3-bromo-5,7-dimethyl-l-adamantyl]methyl]pyrazole
To the solution of the product from Step C (17.88 g, 55.3 mmol) in THF (277 mL) was added butyllithium (2.5 M in THF, 66 mL, 3 eq) at -78 °C, then after 1 h, iodomethane (17.2 mL, 5 eq) was added. After 10 min, the reaction mixture was quenched with a saturated solution of NH4CI, 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.
*H NMR (400 MHz, DMSO-d6): 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); 13C NMR (100 MHz, DMSO-d6) 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-ESI (m/z): [M+H]+ calcd for C17H26BrN2 : 337.1279 found: 337.1289.
Step E: 2-[[3,5-dimethyl-7-[(5-methylpyrazol-l-yl)methyl]-l-adamantyljoxy]éthanol
The mixture of the 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 h. After the reaction mixture was diluted with water and extracted with EtOAc, 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 purification.
XH NMR (400 MHz, DMSO-de): 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); 13C NMR (100 MHz, DMSO-d6) 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; HRMS-ESI (m/z): [M+H]+ calcd for C19H31N2O2 : 319.2386 found: 319.2387.
Step F: tert-butyl-diphenyl-[2-[[3,5-dimethyl-7-[(5-methylpyrazol-l-yl)methyl]-ladamantyl]oxy]ethoxy]silane
175
To the 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 teri-butyl-chloro-diphenyl-silane (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) afforded the desired product (27.0 g, 87.8%).
Ή NMR (400 MHz, DMSO-d6): 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); 13C NMR (100 MHz, DMSO-d6) 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-ESI (m/z): [M+H]+ calcd for C35H49N2O2Si : 557.3563 found: 557.3564.
Step G: tert-butyl-diphenyl-[2-[[3-[(4-iodo-5-methyl-pyrazol-l-yl)methyl]-5J-dimethyl-ladamantyl]oxy]ethoxy]silane
To the solution of the product from Step F (27.0 g, 48.56 mmol) in DMF (243 mL) was added AModosuccinimide (13.6 g, 1.25 eq) and the reaction mixture was stirred for 2 h. After dilution with water, the mixture was extracted with DCM. The combined organic layers were washed with saturated solution of sodium-thiosulphate and brine, dried, and concentrated to afford the desired product (30.1g, 90%).
‘H NMR (400 MHz, DMSO-d6): 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); 13C NMR (100 MHz, DMSO-d6) 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; HRMSESI (m/z): [M+H]+ calcd for C35H48IN2O2Si: 683.2530 found: 683.2533.
Step H: tert-butyl-diphenyl-[2-[[3,5-dimethyl-7-[[5-methyl-4-(4,4,5,5-tetraniethyl-l,3,2dioxaborolan-2-yl)pyrazol-l-yl]methyl]-l-adamantyl]oxy]ethoxy]silane
To the product from Step G (17.5 g, 25.6 mmol) in THF (128 mL) was added chloro(isopropyl)magnesium-LiCl (1.3 M in THF, 24 mL, 1.2 eq) at 0 °C, stirred for 40 min, treated with 2-isopropoxy-4,4,5,5-tetramethyl-l,3,2-dioxaborolane (15.7 mL, 3 eq), and the reaction mixture was stirred for 10 min. After dilution with a saturated solution NH4CI 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 desired product (15.2 g, 86.9%).
176 φ Ή NMR (400 MHz, DMSO-d6): 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); 13C NMR (100 MHz, DMSO-d6) 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; HRMS-ESI (m/z):
[M+H]+ calcd for C41H60BN2O4Si: 683.4415 found: 683.4423.
Préparation 13: methyl 3-bromo-6-(methylamino)pyridine-2-carboxylate
Step A: methyl 6-[bis(tert-butoxycarbonyl)amino]-3-bromo-pyridine-2-carboxylate
To methyl 6-amino-3-bromo-pyridine-2-carboxylate (25.0 g, 108.2 mmol) and DM AP (1.3 g, 0.1 eq) in DCM (541 mL) was added Boc2O (59.0 g, 2.5 eq) at 0 °C and the reaction mixture 10 was stirred for 2.5 h. After the addition of a saturated solution of NaHCO3 and the extraction with DCM, the combined organic phases were dried and concentrated to get the desired product (45.0 g, 72.3%).
LC/MS (Ci7H23BrN2O6Na) 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) was added TFA (17.1 mL, 3 eq) at 0 °C and the reaction mixture was stirred for 18 h. After washing with a saturated solution of NaHCO3 and brine, the combined organic phases were dried, concentrated, and purified by column chromatography (silica gel, heptane and EtOAc as eluents) to give the desired product (28.3 g, 115.2%).
Hl NMR (400 MHz, DMSO-d6): δ ppm 10.29 (s, 1H), 8.11 (d, 1H), 7.88 (d, 1H), 3.87 (s, 3H), 1.46 (s, 9H) 13C NMR (100 MHz, DMSO-d6) δ ppm 165.6, 153.1, 151.8/148.3, 143.5, 116.3, 109.2, 53.2, 28.4. LC/MS (Ci2Hi5BrN2O4Na) 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 Cs2CO3 (8.7 g, 25 3 eq) and iodomethane (0.67 mL, 1.2 eq) and the reaction mixture was stirred for 3 h. After dilution with water and extraction with EtOAc, the combined organic phases were washed with brine, dried and concentrated to give the desired product (3.5 g, 112%).
Ή NMR (400 MHz, DMSO-d6): δ ppm 8.13 (d, 1H), 7.78 (d, 1H), 3.90 (s, 3H), 3.27 (s, 3H), 1.47 (s, 9H); 13C NMR (100 MHz, DMSO-d6) δ ppm 165.5,153.6, 153.6,147.5, 142.8, 122.5,
177
111.3, 82.0, 53.3, 34.3, 28.2; HRMS-ESI (m/z): [M+H]+ calcd for C13H18BrN2O4: 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 h. Purification by column chromatography (silica gel, heptane and EtOAc as eluents) afforded the desired product (2.1 g, 96%).
Ή NMR (400 MHz, DMSO-d6): δ ppm 7.63 (d, 1H), 7.04 (q, 1H), 6.53 (d, 1H), 3.83 (s, 3H), 2.73 (d, 3H); 13C NMR (100 MHz, DMSO-d6) δ ppm 166.6,158.2, 148.2,141.3, 112.1, 101.3, 52.9, 28.3; HRMS-ESI (m/z): [M]+ calcd for C8H9BrN2O2: 243.9847 found: 243.9843.
Préparation 14: methyl 3-[l-[[3,5-dimethyl-7-[2-(p-tolylsulfonyIoxy)ethoxy]-ladamantyl] methyl] -5-methyl-pyrazol-4-yI] -6- [methyl- [5-methyl-6- [(Z) - [3 - (2trimethylsilylethoxymethyl)-l,3-benzothiazol-2-ylidene]amino]pyridazin-3yl]amino]pyridine-2-carboxylate
Step A: methyl 3-[l-[[3-[2-[tert-butyl(diphenyl)silyl]oxyethoxy]-5,7-dimethyl-Iadamantyl]methyl]-5-methyl-pyrazol-4-yl]-6-(methylamino)pyridine-2-carboxylate
The mixture of the product from Préparation 13 (2.07 g, 8.45 mmol), the product from Préparation 12 (6.9 g, 1.2 eq), CS2CO3 (8.26 g, 3 eq), and Pd(AtaPhos)2C12 (374 mg, 0.1 eq) in 1,4-dioxane (51 mL) and water (8.5 mL) was stirred at 80 °C for 1 h. Purification by column chromatography (silica gel, heptane and EtOAc as eluents) afforded the desired product (4.5 g, 74%).
Ή NMR (400 MHz, DMSO-d6): 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); 13C NMR (100 MHz, DMSO-d6) 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-ESI (m/z): [M+H]+ calcd for C43H57N4O4Si: 721.4149 found: 721.4148.
Step B: methyl 3-[l-[[3-[2-[tert-butyl(diphenyl)silyl]oxyethoxy]-5,7-dimethyl-ladamantyl]methyl]-5-methyl-pyrazol-4-yl]-6-[methyl-[5-methyl-6-[(Z)-[3-(2trimethylsilylethoxymethyl)-l,3-benzothiazol-2-ylidene]amino]pyridazin-3yl]amino]pyridine-2-carboxylate
Using Buchwald General Procedure II starting from the product from Step A at reflux for 18
178 h, 4.7 g (86%) of the desired product was obtained.
Ή NMR (400 MHz, DMSO-dô): 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); HRMSESI (m/z): [M+H]+ calcd for C61H79N8O5SSi2: 1091.5433 found: 1091.5426,
Step C: methyl 3-[l -[[3-(2-hydroxyethoxy) -5,7-dimethyl-l -adamantyl]methyl]-5methyl-pyrazol-4-yl]-6-[methyl-[5-methyl-6-[(Z)-[3-(2-trimethylsilylethoxymethyl)-l,3benzothiazol-2-ylidene]amino]pyridazin-3-yl]amino]pyridine-2-carboxylate
To the product from Step B (1.0 g, 0.916 mmol) in THF (9 mL) was added a 1 M solution of TB AF in THF (1.0 mL, 1.1 eq) at 0 °C and the reaction mixture was stirred for 1 h. After quenching with a saturated solution of NH4C1 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 desired product (752 mg, 96%).
Ή NMR (500 MHz, dmso-d6) 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); HRMS-ESI (m/z): [M+H]+ calcd for C45H61N8O5SSi: 853.4255 found: 853.4256.
Step D:methyl 3-[l-[[3,5-dimethyl-7-[2-(p-tolylsulfonyloxy)ethoxy]-l-adamantyl]methyl]-5methyl-pyrazol-4-yl]-6-[methyl-[5-methyl-6-[(Z)-[3-(2-trimethylsilylethoxymethyl)-l,3benzothiazol-2-ylidene]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 mixture was stirred for 1 h. Purification by column chromatography (silica gel, heptane and EtOAc as eluents) afforded the desired product (722 mg, 81%).
’H NMR (400 MHz, DMSO-dô): 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); 13C NMR (100 MHz, DMSO-dô) δ ppm 141.2,137.5,130.6,128.1,127.2, 123.4,123.4,123.1,
179
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; HRMSESI (m/z): [M+H1+ calcd for C„H,7NsO7S7Si: 1007.4343 found: 1007.4344. x z L J 32 O / o l Δ
180
Example 1: 2-({6-[(l,3-Benzothiazol-2-yl)amino]-4,5-dimethylpyridazin-3-yl}amino)l,3-thiazole-4-carboxylic acid
Step A: ethyl 2-({6-[(l ,3-benzothiazol-2-yl)amino]-4,5-dimethylpyridazin-3yl}amino)-l ,35 thiazole-4-carboxylate
To an oven-dried microwave vial was added ethyl 2-aminothiazole-4-carboxylate (71 mg, 0.41 mmol, 1.2 eq), the product from Préparation 9a (100 mg, 0.34 mmol, 1 eq), Xantphos (19.9 mg, 0.03 mmol, 0.1 eq), tris(dibenzylideneacetone)dipalladium(0) (15.8 mg, 0.02 mmol, 0.05 eq) and césium carbonate (134.5 mg, 0.41 mmol, 1.2 eq). The vessel was evacuated and flushed 10 with nitrogen (x3), and then toluene (6 mL) was added. The mixture was sparged with nitrogen (10 mins) 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 (magnésium sulfate) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 12 g RediSep™ silica cartridge) eluting with a gradient of 0 15 - 100% ethyl acetate in Ao-heptane afforded the desired product (63 mg, 0.15 mmol, 43%).
LC/MS (Ci9H18N6O2S2) 427 [M+H]+; RT 1.32 (LCMS-V-B1) 'FI NMR (400 MHz, DMSO-d6) 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-[(l,3-benzothiazol-2-yl)amino]-4,5-dimethylpyridazin-3-yl}amino)-l,3thiazole-4-carboxylic 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 2M aqueous lithium hydroxide (90 pL, 0.18 mmol, 2 eq) and the mixture was stirred at ambient température for 30 min, then heated at 40 °C for 1 h, and finally at 80 °C for 1 h. After 25 allowing to cool to ambient température the reaction was partitioned between ethyl acetate (10
181 mL) and water (10 mL) adjusting to pH 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 drying under vacuum afforded the desired product as an orange solid (14 mg, 0.04 mmol, 39%).
HRMS-ESI (m/z) [M+H]+ calcd for Ci7Hi5N6O2S2: 399.0692, found 399.0729.
Example 2: 6-({6-[(l,3-Benzothiazol-2-yl)amino]-4,5-dimethylpyridazm-3yl}amino)pyridine-2-carboxylic acid
Step A: ethyl 6-({6-[(l,3-benzothiazol-2-yl)amino]-4,5-dimethylpyridazin-3yl}amino)pyridine-2-carboxylate
To an oven-dried microwave vial was added ethyl 6-aminopyridine-2-carboxylate (171 mg, 1.03 mmol, 1.5 eq), the product from Préparation 9a (200 mg, 0.69 mmol, 1 eq), Xantphos (39.8 mg, 0.07 mmol, 0.1 eq), tris(dibenzylideneacetone)dipalladium(0) (31.5 mg, 0.03 mmol, 0.05 eq) and césium carbonate (336 mg, 1.03 mmol, 1.5 eq). The vessel was evacuated and flushed with nitrogen (x3), and then toluene (5 mL) was added. The mixture was sparged with nitrogen (10 mins) then heated at 180 °C for 3 h under microwave irradiation. The mixture was diluted with ethyl acetate and water and filtered through a small silica pad. The organic layer was washed with brine, dried (magnésium sulfate), filtered and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 24 g RediS ep™ silica cartridge) eluting with a gradient of 0 - 100% ethyl acetate in hexane afforded the desired compound as a yellow, glassy solid (55 mg, 0.13 mmol, 19%).
LC/MS (C2iH2oN602S) 421 [M+H]+; RT 1.17 (LCMS-V-B1) ‘H NMR (400 MHz, DMSO-d6) 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,
182
I 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).
Ste^B: 6-({6-[l,3-benzothiazol-2-yl)amino]-4,5-dimethylpyridazin-3-yl}amino)pyridine-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 2M aqueous lithium hydroxide (0.12 mL, 0.24 mmol, 2 eq) and the mixture was heated at 90 °C for 1 h. The mixture was cooled to ambient température, the solvent evaporated in vacuo and the residue was triturated with water and collected by filtration to afford the desired product as an orange solid (9 mg, 0.02 mmol, 19%).
HRMS-ESI (m/z) [M+H]+ calcd for Ci9Hi7N6O2S: 393.1128, found 393.1163.
Example 3: 2-({6-[(l,3-Benzothiazol-2-yl)amino]-4,5-dimethylpyridazin-3-yl}amino)-5(3-{4-[3-(dimethylamino)prop-l-yn-l-yI]-2-fluorophenoxy}propyl)-l,3-thiazole-4carboxylic acid
Step A: ethyl2-({6-[(l,3-benzothiazol-2-yl)amino]-4,5-dimethylpyridazin-3-yl}amino)-5-(3· {4-[3-(dimethylamino)prop-l-yn-l-yl]-2-fluorophenoxy}propyl)-l,3-thiazole-4-carboxylate
To an oven-dried microwave vial was added the product from Préparation 9a (376 mg, 1.29 mmol, 1.5 eq), the product from Préparation 3u (350 mg, 0.86 mmol, 1 eq), césium carbonate (394 mg, 1.21 mmol, 1.4 eq), Xantphos (49.9 mg, 0.09 mmol, 0.1 eq) and 20 tris(dîbenzylideneacetone)dipalladium(0) (39.5 mg, 0.04 mmol, 0.05 eq). The vessel was evacuated and flushed with nitrogen (x3), and then trifluorotoluene (10 mL) was added. The mixture was sparged with nitrogen (10 mins) then heated at 180 °C for 1 h under microwave irradiation. Purification by reverse phase automated flash chromatography (CombiFlash Rf, C18 43g RediSep column) eluting with 5 - 95% acetonitrile in water afforded the desired 25 product as a bright yellow glass (266 mg, 0.4 mmol, 47%).
183
LC/MS (C33H34FN7O3S2) 660 [M+H]+; RT 1.18 (LCMS-V-B1)
H NMR (400 MHz, DMSO-d6) δ 11.31 (br s, 1H), 10.81 (br s, 1H), 7.88 (d, J = 7.8 Hz, 1H), 7.50 (d, J = 7.8 Hz, 1H), 7.37 (ddd, J = 8.2, 7.3, 1.3 Hz, 1H), 7.31 (dd, J = 11.9, 2.0 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).
StepJB: 2-({6-[(l,3-benzothiazol-2-yl)amino]-4,5-dimethylpyridazin-3-yl}amino)-5-(3-{4-[3(dimethylamino)prop-l-yn-l-yl]-2-fluorophenoxy}propyl)-l,3-thiazole-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 2M aqueous lithium hydroxide (0.4 mL, 0.81 mmol, 2 eq) and the mixture was stirred at reflux for 3 h. After allowing to cool to ambient température, the reaction was concentrated in vacuo and triturated with water (10 mL). The solids were collected by filtration, washed with water, and dried under vacuum to afford the desired product as a bright yellow solid (209 mg, 0.33 mmol, 82%) [as a lithium sait].
HRMS-ESI (m/z): [M+H]+ calcd for C31H31FN7O3S2: 632.1908, found 632.1914.
Example 4: 2-({6-[(l,3-Benzothiazol-2-yl)amino]pyridazin-3-yl}amino)-l,3-thiazole-4· carboxylic acid
Step A: ethyl 2-({6-[(l,3-benzothiazol-2-yl)amino]pyridazin-3-yl}amino)-l,3-thiazole-4carboxylate
To an oven-dried flask was added the product from Préparation 9b (500 mg, 1.9 mmol, 1 eq), ethyl 2-amino-l,3-thiazole-4-carboxylate (393 mg, 2.28 mmol, 1.2 eq), césium carbonate (930 mg, 2.85 mmol, 1.5 eq), Xantphos (110 mg, 0.19 mmol, 0.1 eq) and tris(dibenzylîdeneacetone)dipalladium(0) (87.1 mg, 0.1 mmol, 0.05 eq). The flask was evacuated and flushed with nitrogen (x3) and then toluene (20 mL) was added. The mixture was then sparged with nitrogen (15 mins) then heated at 180 °C for 1.5 h under microwave irradiation. The mixture was partitioned between dichloromethane (200 mL) and water (200
184 mL) and the organic phase was concentrated in vacuo. The crude material was triturated with dichloromethane and filtered, washed with dichloromethane and dried to afford the desired product as a yellow solid (417 mg, 1.05 mmol, 55%).
LC/MS (C17H14N6O2S2) 397 [M+H]+; RT 1.20 (LCMS-V-B1)
Ή NMR (400 MHz, DMSO-d6) 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-[(l,3-benzothiazol-2-yl)amino]pyridazin-3-yl}amino)-l,3-thiazole-4carboxylic 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 2M aqueous lithium hydroxide (1.05 mL, 2.09 mmol, 2 eq) and the reaction was heated at reflux for 1 h. The mixture was allowed to cool to ambient température, then the solvent was removed in vacuo and the residue was triturated with acetone, followed by a 1:1 mix of isopropanol / water. The solid was collected by filtration and dried in vacuo to afford the desired product as a brown solid (161 mg, 0.43 mmol, 42%).
HRMS-ESI (m/z) [M+H]+ calcd for C15H11N6O2S2: 371.0379, found 371.0404.
Example 5: 2-({6-[(l,3-Benzothiazol-2-yl)amino]-5-(propan-2-yl)pyridazin-3yl}amino)-l,3-thiazole-4-carboxylic acid
Step A: ethyl2-({6-[(l,3-benzothiazol-2-yl)amino]-5-(propan-2-yl)pyridazin-3-yl}amino)l,3-thiazole-4-carboxylate
To an oven-dried microwave vial was added the product from Préparation 9cb (367 mg, 1.2 mmol, 1 eq), ethyl 2-amino-l,3-thiazole-4-carboxylate (249 mg, 1.44 mmol, 1.2 eq), césium carbonate (589 mg, 1.81 mmol, 1.5 eq), Xantphos (69.7 mg, 0.12 mmol, 0.1 eq) and tris(dibenzylideneacetone)dipalladium(0) (55.1 mg, 0.06 mmol, 0.05 eq). The vial was evacuated and flushed with nitrogen (x3) and then toluene (10 mL) was added. The mixture
185 was sparged with nitrogen (15 min) 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 (magnésium sulfate), and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 40 g RediSep™ silica cartridge) eluting with a gradient of 0 - 50% ethyl acetate in Ao-heptane afforded the desired product as a bright yellow solid (303 mg, 0.69 mmol, 57%).
LC/MS (C20H20N6O2S2) 441 [M+H]+; RT 1.39 (LCMS-V-B1) XH NMR (400 MHz, DMSO-d6) 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-[(l,3-benzothiazol-2-yl)amino]-5-(propan-2-yl)pyridazin-3-yl}amino)-l,3thiazole-4-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 2M aqueous lithium hydroxide (0.68 mL, 1.36 mmol, 2 eq) and the reaction heated at 60 °C for 18 h. The mixture was allowed to cool to ambient température and concentrated in vacuo. The residue was diluted with water and acidified with 2M aqueous hydrochloric acid. The résultant solid was collected by filtration, washed with water and dried under vacuum to afford a brown solid (130 mg, 0.32 mmol, 46%) [as a hydrochloric acid sait],
HRMS-ESI (m/z) [M+H]+ calcd for C18H17N6O2S2: 413.0849, found 413.0885.
Example 6: 2-({6-[(l,3-Benzothiazol-2-yl)amino]-4-(propan-2-yI)pyridazin-3yl}amino)-l,3-thiazole-4-carboxylic acid
Step A: ethyl2-({6-[(l,3-benzothiazol-2-yl)amino]-4-(propan-2-yl)pyridazin-3-yl}amino)1,3-thiazole-4-carboxylate
186
To an oven-dried flask was added the product from Préparation 9ca (500 mg, 1.64 mmol, 1 eq), ethyl 2-amino-l,3-thiazole-4-carboxylate (339 mg, 1.97 mmol, 1.2 eq), Xantphos (94.9 mg, 0.16 mmol, 0.1 eq), césium carbonate (802 mg, 2.46 mmol, 1.5 eq) and tns(dibenzylideneacetone)dipalladium(0) (75.1 mg, 0.08 mmol, 0.05 eq). The vial was 5 evacuated and flushed with nitrogen (x3) and then toluene (20 mL) was added. The mixture was sparged with nitrogen (10 min) then heated at 200 °C for 1.5 h under microwave irradiation. Further Xantphos (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 sparged with nitrogen (10 min) then heated at 180 °C for 3 h under microwave irradiation. The reaction was partitioned between 10 ethyl acetate and water, the aqueous phase extracted with ethyl acetate, and the combined organics washed with brine, dried (magnésium sulfate) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 40 g RediSep™ silica cartridge) eluting with a gradient of 0 - 55% ethyl acetate in Ao-heptane afforded the desired product as an orange solid (147 mg, 0.33 mmol, 20%).
LC/MS (C20H20N6O2S2) 441 [M+H]+; RT 1.35 (LCMS-V-B1)
Step B: 2-({6-[(l,3-benzothiazol-2-yl)amino]-4-(propan-2-yl)pyridazin-3-yl}amino)-l,3thiazole-4-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 2M aqueous lithium hydroxide (0.33 mL, 0.67 mmol, 2 eq) and mixture was heated at 20 reflux for 1 h. The mixture was cooled to ambient température and concentrated in vacuo. The residue was taken up in water and acidified to pH 6. The aqueous phase was extracted with ethyl acetate, and the organics were dried (magnésium sulfate) and concentrated in vacuo. Purification by reverse phase automated flash chromatography (CombiFlash Rf, C18 4.3g RediSep column) eluting with a gradient of 5 - 95% acetonitrile in water afforded the desired 25 product as a yellow solid (7 mg, 0.02 mmol, 5%).
HRMS-ESI (m/z) [M+H]+ calcd for C18H17N6O2S2: 413.0849, found 413.0886.
Example 7: 2-({6-[(l,3-Benzothiazol-2-yl)ammo]-5-methylpyridazin-3-yl}amino)-l,3thiazoIe-4-carboxylic acid
187
H
Step A: ethyl 2-({6-[(l ,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}amino)-l ,3thiazole-4-carboxylate
To an oven-dried microwave vial was added the product from Préparation 9db (400 mg, 1.45 5 mmol, 1 eq), ethyl 2-amino-l,3-thiazole-4-carboxylate (373 mg, 2.17 mmol, 1.5 eq), césium carbonate (706 mg, 2.17 mmol, 1.5 eq), tris(dibenzylîdeneacetone)dipalladium(0) (66.2 mg, 0.07 mmol, 0.05 eq) and Xantphos (83.6 mg, 0.14 mmol, 0.1 eq). The vessel was evacuated and flushed with nitrogen (x3), and then toluene (15 mL) was added. The mixture was sparged with nitrogen (5 min) then heated at 150 °C for 2 h under microwave irradiation. Further ethyl 210 amino-l,3-thiazole-4-carboxylate (375 mg, 2.17 mmol, 1.5 eq), Xantphos (83.6 mg, 0.14 mmol, 0.1 eq), and tris(dîbenzylideneacetone)dipalladium(0) (66.2 mg, 0.07 mmol, 0.05 eq) were added, the mixture was sparged with nitrogen (5 min) then heated at 180 °C for 1 h under microwave irradiation. The mixture was concentrated in vacuo and purification by reverse phase automated flash chromatography (CombiFlash Rf, C18 4.3g RediSep column) eluting 15 with a gradient of 5 - 95% acetonitrile in water afforded the desired product (169 mg, 0.41 mmol, 28%).
LC/MS (C18H16N6O2S2 ) 413 [M+H]+; RT 1.27 (LCMS-V-B1)
Ή NMR (400 MHz, DMSO-d6) 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 = 20 7.1 Hz, 2H), 2.40 (s, 3H), 1.32 (t, J = 7.1 Hz, 3H).
Step B: 2-({6-[(l,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}amino)-l,3-thiazole-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 2M aqueous lithium hydroxide (0.49 mL, 0.49 mmol, 1.2 eq) and the mixture 25 was stirred at 100 °C for 4.5 h. The reaction mixture was allowed to cool to ambient température and concentrated in vacuo. The crude product was suspended in a 2:1 mixture of isopropanol /
188 water and stirred for 30 min. The solids were collected by filtration and dried under vacuum to afford the desired product as a bright yellow solid (17 mg, 0.04 mmol, 11%) [as a lithium sait].
HRMS-ESI (m/z) [M+H]+ calcd for C16H13N6O2S2: 385.0536, found 385.0567.
Example 8: 2-({6-[(l,3-Benzothiazol-2-yl)amino]-5-methyl-4-(propan-2-yl)pyridazin-3yl}amino)-l,3-thiazole-4-carboxylic acid
N S
Step A: ethyl2-({6-[(l,3-benzothiazol-2-yl)amino]-5-methyl-4-(propan-2-yl)pyridazin-3yl}amino)-l,3-thiazole-4-carboxylate
To an oven-dried microwave vial was added the product from Préparation 9ea (300 mg, 0.94 mmol, 1 eq), ethyl 2-amino-l,3-thiazole-4-carboxylate (243 mg, 1.41 mmol, 1.5 eq), césium carbonate (460 mg, 1.41 mmol, 1.5 eq), Xantphos (218 mg, 0.38 mmol, 0.4 eq), and trîs(dibenzylideneacetone)dipalladium(0) (172.4 mg, 0.18 mmol, 0.2 eq). The vessel was evacuated and flushed with nitrogen (x3), and then toluene (4 mL) was added. The mixture was sparged with nitrogen (15 mins) 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 4.3g RediSep column) eluting with a gradient of 5 - 95% acetonitrile in water afforded the desired product (22 mg, 0.05 mmol, 5%).
LC/MS (C21H22N6O2S2) 455 [M+H]+; RT 1.42 (LCMS-V-B1) *H NMR (400 MHz, DMSO-d6) 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-[(l,3-benzothiazol-2-yl)amino]-5-methyl-4-(propan-2-yl)pyridazin-3yl}amino)-1,3-thiazole-4-carboxylic acid
189
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 2M aqueous lithium hydroxide (50 pL, 0.1 mmol, 2 eq) and the mixture was heated at 75 °C for 1 h. The reaction mixture was allowed to cool to ambient température and concentrated in vacuo. The résultant solid was taken back up in water (2 mL) and the pH was adjusted to 6. The organics were extracted with ethyl acetate (20 mL), dried (magnésium sulfate), concentrated in vacuo and dried under vacuum to afford the desired product as a light orange solid (19 mg, 0.04 mmol, 92%).
HRMS-ESI (m/z) [M+H]+ calcd for C19H19N6O2S2: 427.1005, found 427.1043.
Example 9: 2-({6-[(l,3-Benzothiazol-2-yl)ammo]-4,5-dimethylpyridazin-3-yl}amino)-5(3-phenoxypropyl)-l,3-thiazole-4-carboxylic acid
Step A: ethyl2-({6-[(l,3~benzothiazol-2-yl)amino]-4,5-dimethylpyridazin-3-yl}amino)-5-(3phenoxypropyl)-l,3-thiazole-4-carboxylate
To an oven-dried microwave vial was added the product from Préparation 9a (45.7 mg, 0.16 mmol, 1 eq), the product from Préparation 3v (48.2 mg, 0.16 mmol, 1 eq), Xantphos (9.1 mg, 0.02 mmol, 0.1 eq), tris(dibenzylideneacetone)dipalladium(0) (7.2 mg, 0.01 mmol, 0.05 eq) and césium carbonate (61.5 mg, 0.19 mmol, 1.2 eq). The vessel was evacuated and flushed with nitrogen (x3) and then toluene (2 mL) was added. The mixture was sparged with nitrogen (10 mins) then heated at 180 °C for 2 h under microwave irradiation. The reaction was partitioned between ethyl acetate and water, then successively washed with water and brine. The organic phase was dried (magnésium sulfate) and the solvent removed in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 12 g RediSep™ silica cartridge) eluting with a gradient of 0 - 10% methanol in afforded the desired product as a brown glass (19.2 mg, 0.03 mmol, 22%).
LC/MS (C28H28N6O3S2) 561 [M+H]+; RT 1.51 (LCMS-V-B1)
190 XH NMR (400 MHz, DMSO-d6) 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-[(l,3-benzothiazol-2-yl)aminoJ-4,5-dimethylpyridazin-3-yl}amino)-5-(3phenoxypropyl)-l,3-thiazole-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 2M aqueous lithium hydroxide (14.2mg, 0.34 mmol, 10 eq) and the mixture was heated at reflux for 10 h. After allowing to cool to ambient température the solvent was removed in vacuo and the residue was triturated with water and the crude product collected by filtration. Purification by reverse-phase préparative HPLC (method HPLC-V-A2) afforded the desired product as a beige solid (3.5 mg, 6.6 pmol, 19%).
HRMS-ESI (m/z) [M+H]+ calcd for C26H25N6O3S2: 533.1424, found 533.1460.
Example 10: 2-({6-[(l,3-Benzothiazol-2-yl)amino]-4-methyl-5-(propan-2-yl)pyridazin-3yl}amino)-l,3-thiazole-4-carboxylic acid
Step A: ethyl 2-({6-[(l,3-benzothiazol-2-yl)amino]-4-methyl-5-(propan-2-yl)pyridazin-3yl}amino) -1,3-thiazole-4-carboxylate
To an oven-dried microwave vial was added the product from Préparation 9eb (114 mg, 0.36 mmol, 1 eq), ethyl 2-amino-l,3-thiazole-4-carboxylate (92.4 mg, 0.54 mmol, 1.5 eq), césium carbonate (175 mg, 0.54 mmol, 1.5 eq), Xantphos (41.4 mg, 0.07 mmol, 0.2 eq), and tris(dibenzylîdeneacetone)dipalladium(0) (32.7 mg, 0.04 mmol, 0.1 eq). The vessel was evacuated and flushed with nitrogen (x3), and then toluene (4 mL) was added. The mixture was sparged with nitrogen (15 mins) then heated at 180 °C for 1 h under microwave irradiation. The mixture was concentrated in vacuo and purification by reverse phase automated flash chromatography (CombiFlash Rf, C18 4.3g RediSep column) eluting with a gradient of 5 70% acetonitrile in water afforded the desired product as a yellow solid (20 mg, 0.04 mmol, 12%).
191
LC/MS (C21H22N6O2S2) 455 [M+H]+; RT 1.45 (LCMS-V-B1).
Step B: 2-({6-[(l,3-benzothiazol-2-yl)amino]-4-methyl-5-(propan-2-yl)pyridazin-3yl}amino) -l,3-thiazole-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 (0.5 mL) was added lithium hydroxide monohydrate (3.69 mg, 0.09 mmol, 2 eq) and the reaction was heated at reflux for 1 h. The mixture was allowed to cool to ambient température and concentrated in vacuo. The product was taken up in water (3 mL), acidified to pH 6 with IM aqueous hydrochloric acid and extracted with 19:1 dichloromethane / isopropanol. The organic extract was dried (magnésium sulfate) and concentrated in vacuo to afford the desired product as a crisp yellow solid (13.5 mg, 0.03 mmol, 72%).
HRMS-ESI (m/z) [M+H]+ calcd for C19H19N6O2S2: 427.1005, found 427.1043.
Example 11: 2-({6-[(l,3-Benzothiazol-2-yl)amino]-4-methylpyridazin-3-yl}amino)-l,3thiazole-4-carboxylic acid
Step A: ethyl 2-({6-[(l,3-benzothiazol-2-yl)amino]-4-methylpyridazin-3-yl}amino)-l,3thiazole-4-carboxylate
To an oven-dried microwave vial was added the product from Préparation 9da (240 mg, 0.87 mmol, 1 eq), ethyl 2-amino-l,3-thiazole-4-carboxylate (224 mg, 1.3 mmol, 1.5 eq), césium carbonate (424 mg, 1.3 mmol, 1.5 eq), tris(dibenzylideneaœtone)dipalladium(0) (39.7 mg, 0.04 mmol, 0.05 eq) and Xantphos (50.2 mg, 0.09 mmol, 0.1 eq). The vessel was evacuated and flushed with nitrogen (x3), and then toluene (15 mL) was added. The mixture was sparged with nitrogen (5 min) then heated at 180 °C for 3 h under microwave irradiation. The mixture was concentrated in vacuo and purification by reverse phase automated flash chromatography (CombiFlash Rf, C18 26g RediSep column) eluting with a gradient of 5 - 95% acetonitrile in water to afford the desired product as a brown solid (44 mg, 0.11 mmol, 12%).
LC/MS (C18H16N6O2S2) 413 [M+H]+; RT 1.24 (LCMS-V-B1)
192 > Ή NMR (400 MHz, DMSO-d6) 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).
StepBy 2-({6-[(l,3-benzothiazol-2-yl)amino]-4-methylpyridazin-3-yl}amino)-l,3-thiazole-45 carboxylic 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 2M aqueous lithium hydroxide (0.21 mL, 0.21 mmol, 2 eq) and the reaction was heated at reflux for 1 h. The mixture was allowed to cool to ambient température then concentrated in vacuo. The solid was triturated with 4:1 isopropanol / water (4 mL) and stirred for 30 min. The 10 solid was collected by filtration, washed with further cold isopropanol (3 mL) and dried under vacuum to afford the desired product as a brown solid (33 mg, 0.09 mmol, 80%).
HRMS-ESI (m/z) [M+H]+ calcd for C16H13N6O2S2: 385.0536, found 385.0572.
Example 12; 2-({6-[(l,3-Benzothiazol-2-yl)amino]-4,5-dimethylpyridazm-3-yl}amino)-5(3-methoxypropyl)-l,3-thiazole-4-carboxylic acid
Step A: ethyl 2-({6-[(l,3-benzothiazol-2-yl)amino]-4,5-dimethylpyridazin-3-yl}amino)-5-(3methoxypropyl)-l,3-thiazole-4-carboxylate
To an oven-dried microwave vial was added the product from Préparation 9a (260 mg, 0.89 mmol, 1 eq), the product from Préparation 3w (219 mg, 0.89 mmol, 1 eq), Xantphos (51.8 mg, 20 0.09 mmol, 0.1 eq), tris(dibenzylideneacetone)dipalladium(0) (41 mg, 0.04 mmol, 0.05 eq) and césium carbonate (350 mg, 1.07 mmol, 1.2 eq). The vessel was evacuated and flushed with nitrogen (x3), and then toluene (5 mL) was added. The mixture was sparged with nitrogen (10 min) then heated at 180 °C for 1 h under microwave irradiation. The réaction was diluted with ethyl acetate, successively washed with water and brine, then dried (magnésium sulfate) and 25 the solvent removed in vacuo. Purification by reverse phase automated flash chromatography (CombiFlash Rf, C18 13g RediSep column) eluting with 5 — 95% acetonitrile in water afforded the desired product as a yellow glass (72.5 mg, 0.15 mmol, 16%).
193
LC/MS (C23H26N6O3S2) 499 [M+H]+; RT 1.38 (LCMS-V-B1)
Ή NMR (400 MHz, DMSO-d6) 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-[(l,3-benzothiazol-2-yl)amino]-4,5-dimethylpyridazin-3-yl}amino)-5-(3methoxypropyl)-l,3-thiazole-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 heated at reflux for 10 h. The reaction was allowed to cool to ambient température and the solvent removed in vacuo. The résultant residue was triturated with water then collected by filtration, washed with water, and dried under vacuum to afford the desired product as a brown solid (40.9 mg, 0.09 mmol, 60%) [as a lithium sait].
HRMS-ESI (m/z) [M+H]+ calcd for C21H23N6O3S2: 471.1268, found 471.1306.
Example 13: 2-({6-[(l,3-Benzothiazol-2-yl)amino]-4,5-dimethylpyridazin-3-yl}amino)-5[3-(2-fluorophenoxy)propyl]-l,3-thiazole-4-carboxylic acid
Step A: ethyl2-({6-[(l,3-benzothiazol-2-yl)amino]-4,5-dimethylpyridazin-3-yl}amino)-5-[3(2-fluorophenoxy)propyl]-l,3-thiazole-4-carboxylate
To an oven-dried microwave vial was added the product from Préparation 9a (162 mg, 0.56 mmol, 1 eq), the product from Préparation 3x (181 mg, 0.56 mmol, 1 eq), Xantphos (36.0 mg, 0.06 mmol, 0.11 eq), tris(dibenzylideneacetone)dipalladium(0) (25.6 mg, 0.03 mmol, 0.05 eq) and césium carbonate (243 mg, 0.75 mmol, 1.34 eq). The vessel was evacuated and flushed with nitrogen (x3), and then toluene (4 mL) and dimethylformamide (1 mL) were added. The mixture was sparged with nitrogen (10 min) then heated at 180 °C for 1 h under microwave irradiation. The reaction was diluted with ethyl acetate, successively washed with water and brine, dried (magnésium sulfate), and concentrated in vacuo. Purification by reverse phase
194 automated flash chromatography (CombiFlash Rf, C18 13g RediSep column) eluting with a gradient of 5 - 95% acetonitrile in water afforded the desired product as a brown glass (76.1 mg, 0.13 mmol, 24%).
LC/MS (C28H27FN6O3S2) 579 [M+H]+; RT 1.49 (LCMS-V-B1) XH NMR (400 MHz, DMSO-d6) 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-[(l,3-benzothiazol-2-yl)amino]-4,5-dimethylpyridazin-3-yl}amino)-5-[3-(2fluorophenoxy)propyl]-l,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 heated at reflux for 8h. The mixture was allowed to cool to ambient température and the solvent was removed in vacuo. Purification by reverse phase automated flash chromatography (CombiFlash Rf, C18 13g RediSep column) eluting with a gradient of 5 - 95% acetonitrile in water afforded material that was further purified by reverse phase préparative HPLC (method HPLC-V-A2) to afford the desired product as a beige solid (7.2 mg, 0.01 mmol, 10%).
HRMS-ESI (m/z) [M+H]+ calcd for C26H24FN6O3S2: 551.1330, found 551.1368.
Example 14: 2-({6-[(l,3-Benzothiazol-2-yl)amino]-4,5-dimethylpyridazin-3-yl}amino)-5{3-[4-(dimethylcarbamoyl)-2-fluorophenoxy]propyl}-l,3-thiazole-4-carboxylic acid
Step A: ethyl2-({6-[(l,3-benzothiazol-2-yl)amino]-4,5-dimethylpyridazin-3-yl}amino)-5-{3[4-(dimethylcarbamoyl) -2-fluorophenoxy]propyl}-l,3-thiazole-4-carboxylate
To an oven-dried microwave vial was added the product from Préparation 9a (143 mg, 0.49 mmol, 1 eq), the product from Préparation 3y (199 mg, 0.49 mmol, 1 eq), Xantphos (28.5 mg, 0.05 mmol, 0.1 eq), trîs(dibenzylideneacetone)dipalladium(0) (22.5 mg, 0.02 mmol, 0.05
195 eq) and césium carbonate (192 mg, 0.59 mmol, 1.2 eq). The vessel was evacuated and flushed with nitrogen (x3), and then toluene (4 mL) and dimethylformamide (1 mL) were added. The mixture was sparged with nitrogen (10 min) then heated at 180 °C for 1 h under microwave irradiation. The reaction was diluted with ethyl acetate then washed with water followed by brine. The organic extract was dried (magnésium sulfate) and the solvent removed in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 12 g RediSep™ silica cartridge) eluting with a gradient of 0 - 10% methanol in dichloromethane afforded the desired product as a brown glass (60 mg, 0.09 mmol, 19%).
LC/MS (C31H32FN7O4S2) 650 [M+H]+; RT 1.36 (LCMS-V-B1)
Hl NMR (400 MHz, DMSO-d6) 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-[(l,3-benzothiazol-2-yl)amino]-4,5-dimethylpyridazin-3-yl}amino)-5-{3-[4(dimethylcarbamoyl)-2-fluorophenoxy]propyl}-l,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 heated at reflux for 14 h. The reaction was allowed to cool to ambient température and the solvent removed in vacuo. The residue was triturated with water, followed by éthanol, then purified by reverse phase préparative HPLC (method HPLC-V-A2) to afford the desired product as a brown solid (11.6 mg, 0.02 mmol, 20%).
HRMS-ESI (m/z) [M+H]+ calcd for C29H29FN7O4S2: 622.1701, found 622.1740.
Example 15: 2-({6-[(l,3-Benzothiazol-2-yl)amino]-4-ethyl-5-methylpyridazin-3yl}amino)-l,3-thiazoIe-4-carboxylic acid
Step A: ethyl2-({6-[(l,3-benzothiazol-2-yl)amino]-4-ethyl-5-methylpyridazin-3-yl}amino)1,3-thiazole-4-carboxylate
196
To an oven-dried microwave vial was added the product from Préparation 9fa (400 mg, 1.31 mmol, 1 eq), ethyl 2-amino-l,3-thiazole-4-carboxylate (316 mg, 1.84 mmol, 1.4 eq), césium carbonate (599 mg, 1.84 mmol, 1.4 eq), Xantphos (75.9 mg, 0.13 mmol, 0.1 eq) and tris(dibenzylideneacetone)dipalladium(0) (60.1 mg, 0.07 mmol, 0.05 eq). The vial was evacuated and flushed with nitrogen (x3), and then toluene (15 mL) was added. The mixture was sparged with nitrogen (10 mins) then heated at 200 °C for 2 h under microwave irradiation. The reaction was concentrated in vacuo and purification by reverse phase automated flash chromatography (CombiFlash Rf, C18 13g RediSep column) eluting with a gradient of 5 - 95% acetonitrile in water afforded the desired product (70 mg, 0.16 mmol, 12%).
LC/MS (C20H20N6O2S2) 441 [M+H]+; RT 1.37 (LCMS-V-B1) XH NMR (400 MHz, DMSO-d6) δ 11.08 (s, 1H), 10.88 (br s, 1H), 7.94 (s, 1H), 7.89 (br s, 1H), 7.53 (br s, 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-[(l,3-benzothiazol-2-yl)amino]-4-ethyl-5-methylpyridazin-3-yl}amino)-l,3thiazole-4-carboxylic 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 heated at reflux for 1 h. The reaction was allowed to cool to ambient température and concentrated in vacuo. The residue was then suspended in water and the pH adjusted to 6 then re-concentrated. The solid was triturated with isopropanol plus a few drops of water, then collected by filtration, washew with isopropanol, and dried under vacuum to afford the desired product as a brown solid (38.8 mg, 0.09 mmol, 59%).
HRMS-ESI (m/z) [M-H]- calcd for C18H15N6O2S2: 411.0703, found 411.0707.
Example 16: 2-({6-[(l,3-Benzothiazol-2-yl)amino]-5-ethyl-4-methylpyridazin-3yl}amino)-l,3-thiazole-4-carboxylic acid
197
Step A: ethyl2-({6-[(l,3-benzothiazol-2-yl)amino]-5-ethyl-4-methylpyridazin-3-yl}amino)l,3-thiazole-4-carboxylate
To an oven-dried microwave vial was added the product from Préparation 9fb (300 mg, 0.98 mmol, 1 eq), ethyl 2-amino-l,3-thiazole-4-carboxylate (220 mg, 1.28 mmol, 1.3 eq), césium carbonate (449 mg, 1.38 mmol, 1.4 eq), Xantphos (57 mg, 0.1 mmol, 0.1 eq) and tris(dibenzylïdeneacetone)dipalladium(0) (45.1 mg, 0.05 mmol, 0.05 eq). The vial was evacuated and flushed with nitrogen (x3), and then toluene (15 mL) was added. The mixture was sparged with nitrogen (10 mins) then heated at 200 °C for 2 h under microwave irradiation. The reaction was concentrated in vacuo and purification by reverse phase; automated flash chromatography (CombiFlash Rf, C18 13g RediSep column) eluting with 5 -- 95% acetonitrile in water to afford the desired product as a yellow solid (54 mg, 0.12 mmol, 12%).
LC/MS (C20H20N6O2S2) 441 [M+H]+; RT 1.38 (LCMS-V-B1)
Ή NMR (400 MHz, DMSO-d6) 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-[(l,3-benzothiazol-2-yl)amino]-5-ethyl-4-methylpyridazin-3-yl}amino)-l,3thiazole-4-carboxylic 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 2M aqueous lithium hydroxide (0.12 mL, 0.25 mmol, 2 eq) and the reaction was heated at reflux for 2 h. The mixture was concentrated in vacuo and then suspended in water and the pH adjusted to 5-6 before re-concentrating in vacuo. The crude solid was triturated with water, stirred for 30 min, then collected by filtration, washed with cold water and dried under vacuum to afford the desired product as a yellow solid (28.9 mg, 0.07 mmol, 57%).
HRMS-ESI (m/z) [M+H]+ calcd for Ci8Hi7N6O2S2: 413.0849, found 413.0848.
Example 17: 2-[[4-(l,3-Benzothiazol-2-ylamino)-5,6,7,8-tetrahydrophthalazm-lyl]amino]thiazole-4-carboxylic acid
198
Step A: 4-bromo-5,6,7,8-tetrahydrophthalazin-l-amine
The mixture of 4.0 g of l,4-dibromo-5,6,7,8-tetrahydrophthalazine (13.7 mmol) in 50 mL of a 25% solution of NH3 in water was kept at 100 °C for 18 h. After concentration, the residue was 5 taken up in methanol, treated with charcoal, stirred, filtered, concentrated, taken up in EtOAc, washed with a 10% solution of NaOH, dried on MgSO4, filtered, and concentrated to give 1.28 g (41%) of the desired product.
LC-MS-ESI (m/z): [M+H]+ calcd for C8HiiBrN3: 228, 230, found: 228, 230.
Step B: N-(4-bromo-5,6,7,8-tetrahydrophthalazin-l-yl)-l,3-benzothiazol-2-amine
The mixture of 1.05 g of the product from Step A (4.6 mmol, 1 eq.), 1.8 g: of 2-bromo-l,3benzothiazole (8.4 mmol, 1.8 eq.), 736 mg of sodium hydride (60% suspension, 18.4 mmol, 3.6 eq.) in 80 mL of 1,4-dioxane was kept at reflux for 0.5 h. After cooling and quenching with methanol, the mixture was concentrated, treated with water, filtered off and dried. The solid was triturated in EtOAc, filtered off and dried to give 1.56 g (94%) of the desired product.
Ή NMR (500 MHz, DMSO+fc) 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); 13C NMR (125 MHz, dmso-d6) δ ppm 126.6, 122.9, 122.2, 28.8/24, 21.4/21.1; HRMS-ESI (m/z): [M+H]+ calcd for Ci5Hi4BrN4S: 361.0117, found: 361.0107.
Step C: ethyl 2-[[4-(l,3-benzothiazol-2-ylamino)-5,6,7,8-tetrahydrophthalazin-l20 yl]amino]thiazole-4-carboxylate
The mixture of 120 mg of the product from Step B (0.33 mmol, 1 eq.), 57 mg of ethyl 2aminothiazole-4-carboxylate (0.33 mmol, 1 eq.), 12 mg of Pd2(dba)2 (0.013 mmol, 0.04 eq.), 19 mg of Xantphos (0.033 mmol, 0.1 eq.), and 217 mg of Cs2CO3 (0.66 mmol, 2 eq.) in 10 mL of trifluoromethylbenzene was kept at 200 °C for 0.5 h. The reaction mixture was concentrated 25 and purified by préparative HPLC to give 31 mg (21%) of the desired product.
199 φ Ή NMR (500 MHz, DMSO-d6) 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); 13C NMR (125 MHz DMSO-î/6) δ ppm 126.3, 122.5, 121.5; HRMS-ESI (m/z): [M+H]+ calcd for C21H21N6O2S2: 453.1162, found: 453.1151.
Step D: 2-[[4-(l ,3-benzothiazol-2-ylamino)-5,6,7,8-tetrahydrophthalazin-lyl]amino]thiazole-4-carboxylic acid
The mixture of 31 mg of the product from Step C (0.069 mmol, 1 eq.), 5.8 mg of LiOHxH2O (2 eq.), and 8 drops of water in 10 mL of 1,4-dioxane was kept at reflux for 2 h. The reaction mixture was concentrated and purified by préparative HPLC to give 24 mg (83%) of the 10 desired product.
HRMS-ESI (m/z): [M+H]+ calcd for C19H17N6O2S2: 425.0849, found: 425.0854.
Example 18: 2-({6-[(l,3-Benzothiazol-2-yl)amino]-4-(hydroxymethyl)-5·· methylpyridazin-3-yl}amino)-l,3-thiazole-4-carboxylic acid
Step A: ethyl 2-({6-[(l,3-benzothiazol-2-yl)amino]-4-[(benzyloxy)methyl]-5-methylpyridin3-yl}amino)-l,3-thiazole-4-carboxylate
To an oven-dried microwave vial was added the product from Préparation 9ga (176 mg, 0.44 mmol, 1 eq), ethyl 2-amino-l,3-thiazole-4-carboxylate (115 mg, 0.67 mmol, 1.5 eq), césium 20 carbonate (202 mg, 0.62 mmol, 1.4 eq), tris(dibenzylideneacetone)dipalladium(0) (40.6 mg, 0.04 mmol, 0.1 eq) and Xantphos (12.8 mg, 0.02 mmol, 0.05 eq). The vial was evacuated and flushed with nitrogen (x3), and then toluene (15 mL) was added. The mixture was sparged with nitrogen (5 mins) then heated at 180 °C for 1 h under microwave irradiation. The mixture was concentrated in vacuo and purification by reverse phase automated flash chromatography 25 (CombiFlash Rf, C18 24g RediSep column) eluting with a gradient of 5 - 95% acetonitrile in water to afford the desired product (82 mg, 0.15 mmol, 35%).
200
LC/MS (C26H24N6O3S2) 533 [M+H]+; RT 1.44 (LCMS-V-B1)
Ή NMR (400 MHz, DMSO-d6) 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: ethyl2-({6-[(l,3-benzothiazol-2-yl)amino]-4-(hydroxymethyl)-5-methylpyridin-3yl}amino)-l,3-thiazole-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 IM boron trichloride solution (0.68 mL, 0.68 mmol, 5 eq) was added dropwise. The mixture was then allowed to warm to ambient température over 2 h. The reaction mixture was quenched by addition of methanol and concentrated in vacuo. The résultant solid was triturated with dichloromethane, filtered, washed with dichloromethane and dried under vacuum to afford the desired product as a cream solid (54 mg, 0.12 mmol, 90%).
LC/MS (C19H18N6O3S2) 443 [M+H]+; RT 1.23 (LCMS-V-B1) ‘H NMR (400 MHz, DMSO-d6) 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-[(l,3-benzothiazol-2-yl)amino]-4-(hydroxymethyl)-5-methylpyridin-3yl}amino)-l,3-thiazole-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 2M aqueous lithium hydroxide (0.56 mL, 0.56 mmol, 5 eq) and the mixture was heated at reflux for 30 min. The mixture was cooled to ambient température and concentrated in vacuo. The solid was taken up in water (5 mL) and neutralised to pH 6 with IM HCl and then concentrated in vacuo. The solid was then triturated with 4:1 isopropanol / water and stirred for 30 min. The solids were collected by filtration and dried under vacuum to afford the desired product as an orange solid (30 mg, 0.07 mmol, 64%).
HRMS-ESI (m/z) [M+H]+ calcd for C17H15N6O3S2: 415.0642, found 415.0638.
Example 19: 2-({6-[(l,3-Benzothiazol-2-yl)amino]-4-cycIopropyl-5-methylpyridazin-3yl}amino)-l,3-thiazole-4-carboxylic acid
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Step A: ethyl 2-({6-[(l,3-benzothiazol-2-yl)amino]-4-cyclopropyl-5-methylpyridazin-3yl}amino)-1,3-thiazole-4-carboxylate
To an oven-dried microwave vial was added the product from Préparation 9ha (300 mg, 0.95 5 mmol, 1 eq), ethyl 2-amino-l,3-thiazole-4-carboxylate (245 mg, 1.42 mmol, 1.5 eq), césium carbonate (463 mg, 1.42 mmol, 1.5 eq), Xantphos (54.8 mg, 0.09 mmol, 0.1 eq) and tris(dîbenzylideneacetone)dipalladium(0) (43.4 mg, 0.05 mmol, 0.05 eq). The vial was evacuated and flushed with nitrogen (x3), and then alpha,alpha,alpha-triüuorotohiene (10 mL) was added. The mixture was sparged with nitrogen (5 mins) then heated at 180 °C for 1 h under 10 microwave irradiation. The reaction mixture was concentrated in vacuo and purification by reverse phase automated flash chromatography (CombiFlash Rf, C18 26g RediSep column) eluting with a gradient of 5 - 95% acetonitrile (with 0.08% formic acid modifier) in water (with 0.08% formic acid modifier) afforded the desired product as a bright yellow solid (67 mg, 0.15 mmol, 16%).
LC/MS (C21H20N6O2S2) 453 [M+H]+; RT 1.34 (LCMS-V-B1)
Ή NMR (400 MHz, DMSO-d6) 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-[(l,3-benzothiazol-2-yl)aminoJ-4-cyclopropyl-5-methylpyridazin-3-yl}amino)20 l,3-thiazole-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 2M aqueous lithium hydroxide (0.15 mL, 0.3 mmol, 2 eq) and the mixture was heated at reflux for 1 h. The reaction was concentrated in vacuo, then taken-up in water and the pH was adjusted to 6 with IN aqueous hydrochloric acid, before re-concentrating in vacuo. 25 Trituration with 5:1 isopropanol / water (5 mL) gave a solid that was collected by filtration, washed with isopropanol and dried under vacuum to afford the desired product as a yellow solid (28 mg, 0.07 mmol, 45%).
202
HRMS-ESI (m/z) [M+H]+ calcd for C19H17N6O2S2: 425.0849, found 425.0857.
Example 20: 2-({6-[(l,3-Benzothiazol-2-yl)amino]-5-cyclopropyl-4-methylpyridazin-3yl}amino)-l,3-thiazole-4-carboxylic acid
Step A: ethyl2-({6-[(l,3-benzothiazol-2-yl)amino]-5-cyclopropyl-4-methylpyridazin-3yl}amino)-l,3-thiazole-4-carboxylate
To an oven-dried microwave vial was added the product from Préparation 9hb (219 mg, 0.69 mmol, 1 eq), ethyl 2-amino-l,3-thiazole-4-carboxylate (179 mg, 1.04 mmol, 1.5 eq), césium carbonate (338 mg, 1.04 mmol, 1.5 eq), Xantphos (40 mg, 0.07 mmol, 0.1 eq) and trîs(dibenzylideneacetone)dipalladium(0) (31.7 mg, 0.03 mmol, 0.05 eq). The vial was evacuated and flushed with nitrogen (x3), and then alpha,alpha,«Zp/za-trifluorotolucne (15 mL) was added. The mixture was sparged with nitrogen (5 mins) then heated at 170 °C for 1 h under microwave irradiation. The reaction mixture was concentrated in vacuo and purification by reverse phase automated flash chromatography (CombiFlash Rf, C18 24g RediSep column) eluting with a gradient of 5 - 95% acetonitrile (containing 0.08% formic acid) in water (containing 0.08% formic acid) afforded the desired product (54 mg, 0.12 mmol, 17%).
LC/MS (C2iH20N6O2S2) 453 [M+H]+; RT 1.16 (LCMS-V-B1) ’H NMR (400 MHz, DMSO-d6) 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-[(I,3-benzothiazol-2-yl)amino]-5-cyclopropyl-4-methylpyridazin-3-yl}amino)l,3-thiazole-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 2M aqueous lithium hydroxide (0.24 mL, 0.24 mmol, 2 eq) and the mixture was heated
203 at reflux for 1 h. The reaction was allowed to cool to ambient température and concentrated in vacuo. The residue was taken up in water and neutralised to pH 6 with IM aqueous hydrochloric acid and then concentrated in vacuo. The solid was triturated in 4:1 isopropanol / water and stirred for 30 min. The solids were collected by filtration, washed with cold isopropanol and dried under vacuum to afford the desired product as an orange solid (30 mg, 0.07 mmol, 59%).
HRMS-ESI (m/z) [M+H]+ calcd for C19H17N6O2S2: 425.0849, found 425.0853.
Example 21: 2-({6-[(l,3-Benzothiazol-2-yl)amino]-5-methylpyridazin-3yl}(methyl)amino)-l,3-thiazole-4-carboxylic acid
Step A: ethyl2-[(6-chloro-5-methylpyridazin-3-yl)(methyl)amino]-l ,3-thiazole-4carboxylate
Sodium hydride (60% dispersion, 240 mg, 6 mmol, 1.18 eq) was added slowly to a solution of 3,6-dichloro-4-methylpyridazine (850 mg, 5.21 mmol, 1.02 eq) and ethyl 2-(methylamino)-l,3thiazole-4-carboxylate (950 mg, 5.1 mmol, 1 eq) in tetrahydrofuran (25 mL) at 0 °C under a nitrogen atmosphère. The mixture was stirred for 1 h at 0 °C and for 18 h at ambient température to give a dark brown suspension. Saturated aqueous ammonium chloride solution (75 mL) was added and the mixture extracted with dichloromethane (250 mL). The extracts were washed with water (2 x 75 mL) and brine (75 mL). The solution was dried (magnésium sulfate) and concentrated in vacuo. Purification by flash column chromatography (100 g silica) eluting with 1% methanol in dichloromethane gave a solid that was triturated with diethyl ether (15 mL), filtered, washed with diethyl ether (2 x 10 mL) and dried in vacuo to afford the desired product as a pale brown solid (960 mg, 3.07 mmol, 60%).
LC/MS (C12H13CIN4O2S) 313 [M+H]+; RT 1.17 (LCMS-V-B1) JH NMR (400 MHz, DMSO-d6) 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).
204
Step B: ethyl 2-({6-[(l,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)l,3-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, 0.05 eq) and Xantphos (81.4 mg, 0.14 mmol, 0.1 eq) in 1,4-dioxane (20 mL) under a nitrogen atmosphère and the mixture was stirred. NW-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 h. The solution was allowed to cool to ambient température and concentrated in vacuo. Purification by flash column chromatography (50 g silica), eluting -with ethyl acetate gave a yellow solid that was triturated with methanol (10 mL), filtered, washed with methanol (15 mL) and dried under vacuum to afford the desired product as a yellow solid (315 mg, 0.74 mmol, 53%).
LC/MS (C19H18N6O2S2) 427 [M+H]+; RT 1.32 (LCMS-V-B1) XH NMR (400 MHz, DMSO-d6) 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-[(l,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)-l,3thiazole-4-carboxylic 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 température for 3 hr. Water (10 mL) was added and the solution filtered through celite, the solids were washed with water (5 mL) and the combined filtrate acidified with acetic acid to give a pale yellow suspension. The solids were collected by filtration, washed with water (20 mL) and dried under vacuum to afford the desired product as a pale yellow solid (150 mg, 0.34 mmol, 72%).
LC/MS (C17H14N6O2S2) 399 [M+H]+; RT 1.1 (LCMS-V-B1) XH NMR (400 MHz, DMSO-d6) 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).
205
Example 22: 2-({6-[(l,3-Benzothiazol-2-yl)amino]-4,5-dimethylpyridazm-3-yl}amino)-5(3-{4-[3-(dimethylammo)propyl]-2-fluorophenoxy}propyl)-l,3-thiazole-4-carboxylic acid
Step A: ethyl5-(3-{4-[(lE)-3-(dimethylamino)prop-l-en-l-yl]-2-fluorophenoxy}propyl)-25 acetamido-l,3-thiazole-4-carboxylate
Sodium hydride (60% in minerai oil) (1.61 g, 40.3 mmol, 2.34 eq) was added to a stirred solution of the product from Préparation 6b (4.65 g, 24.1 mmol, 1.4 eq) in dimethylformamide (70 mL). The reaction was stirred at ambient température for 30 min then cooled in an ice bath and a solution of the product from Préparation 3u, Step C (6.57 g, 17.2 mmol, 1 eq) in 10 dimethylformamide (30 mL) was added slowly. After 40 min the reaction was allowed to warm to ambient température and stirred for 5 h. The mixture was cooled, acidified with 2N aqueous HCl and the organics extracted with ethyl acetate, then 3:1 dichloromethane / isopropanol. The combined organics were dried (magnésium sulfate) and the solvent removed in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 80 g RediSep™ 15 silica cartridge) eluting with a gradient of 0 - 10% methanol in dichloromethane afforded the desired product as a brown foam (2.26 g, 5.69 mmol, 33%) that was used in subséquent steps without further purification. A sample of the material was further purified by préparative HPLC (method HPLC-V-A1) to generate analytical data.
LC/MS (C22H28FN3O4S) 450 [M+H]+; RT 0.75 (LCMS-V-B1)
Ή NMR (400 MHz, DMSO-d6) 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-[(lE)-3-(dimethylamino)prop-l-en-l-yl]-225 fluorophenoxy}propyl) -l,3-thiazole-4-carboxylate
206
To a solution of the product from Step A (2.25 g, 4.99 mmol, 1 eq) in éthanol (40 ml ,) was added hydrochloric acid (4M in 1,4-dioxane; 12.5 mL, 49.9 mmol, 10 eq) and the mixture was heated at 60 °C for 18 h. The reaction was allowed to cool to ambient température and the solvent removed in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 40 g RediSep™ silica cartridge) eluting with a gradient of 0 - 20% methanol in dichloromethane afforded a crude product. This was dissolved in methanol and loaded onto a methanol-wet SCX-2 cartridge (10 g). Elution with methanol followed by 3.5N methanolic ammonia, then évaporation afforded the desired product as a brown gum (2.07 g, 5.07 mmol, 100%).
LC/MS (C20H26FN3O3S) 408 [M+H]+; RT 0.71 (LCMS-V-B1) XH NMR (400 MHz, DMSO-d6) 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, 2E1), 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)-l,3thiazole-4-carboxylate
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 added to a flask containing platinum(IV) oxide hydrate, (spatula tip) under a nitrogen atmostphere. The vessel was evacuated and back-filled with nitrogen (x3), then evacuated, placed under an atmosphère of hydrogen, and shaken at ambient température for 72 h. The mixture was filtered through celite (10g pre-packed cartridge), eluted with methanol, and the solvent was removed in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 40 g RediSep™ silica cartridge) eluting with a gradient of 0 - 20% methanol (containing 20% ammonia) in dichloromethane afforded the desired product as a yellow gum (519 mg, 1.27 mmol, 53%).
LC/MS (C20H28FN3O3S) 410 [M+H]+; RT 0.70 (LCMS-V-B1) XH NMR (400 MHz, DMSO-d6) 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-[(l,3-benzothiazol-2-yl)amino]-4,5-dimethylpyridazin-3-yl}amino)-5-(3{4-[3-(dimethylamino)propyl]-2-fluorophenoxy}propyl)-l,3-thiazole-4-carboxylate
207
To an oven-dried microwave vial was added the product from Préparation 9a (118 mg, 0.41 mmol, 1 eq), the product from Step C (200 mg, 0.49 mmol, 1.2 eq), césium carbonate (265 mg, 0.81 mmol, 2 eq), Xantphos (23.6 mg, 0.04 mmol, 0.1 eq) and tris(dibenzylideneacetone)dipalladium(0) (18.6 mg, 0.02 mmol, 0.05 eq). The vial was evacuated and flushed with nitrogen (x3), and then alpha,alpha,(4 mL) was added. The mixture was sparged with nitrogen (5 mins) then heated at 170 °C for 1 h under microwave irradiation. The mixture was partitioned between ethyl acetate and water, and the organic phase dried (magnésium sulfate) and concentrated in vacuo. Purification by reverse phase automated flash chromatography (CombiFlash Rf, C18 13g RediSep column) eluting with 5-95% acetonitrile in water afforded the desired product as a yellow foam (36 mg, 0.05 mmol, 13%).
LC/MS (C33H38FN7O3S2) 664 [M+H]+; RT 1.01 (LCMS-V-B1) XH NMR (400 MHz, DMSO-d6) 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-[(l,3-benzothiazol-2-yl)amino]-4,5-dimethylpyridazin-3-yl}amino)-5-(3-{4-[3(dimethylamino)propyl]-2-fluorophenoxy}propyl)-l,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 heated at reflux for 5.5 h. The mixture was allowed to cool to ambient température and the solvent removed in vacuo. The residue was triturated in water and solids were collected by filtration. The solids were further triturated with diethyl ether, then collected by filtration and dried under vacuum to afford the desired product as a brown solid (19.1 mg, 0.03 mmol, 35%). HRMS-ESI (m/z) [M+H]+ calcd for C31H35FN7O3S2: 636.2221, found 636.2224.
Example 23: 2-[[l-(l,3-Benzothiazol-2-ylamino)-6,7-dihydro-5/fcydopenta[d]pyridazin-4-yI]amino]thiazole-4-carboxylic acid
208
Step A: l,4-dibromo-6,7-dihydro-5H-cyclopenta[d]pyridazine
The mixture of 1.86 g of 2,3,6,7-tetrahydro-l/f-cyclopenta[<f|pyridazine-l,4(5/f)-dione (12.3 mmol, 1 eq.) and 10.55 g (36.8 mmol, 3 eq.) of POBrs in 50 mL of 1,2-dichloroethane was kept 5 at reflux for 3 h. After cooling to rt and concentration, the residue was treated with 100 g of ice, the pH was set to 7 by the addition of cc. NaHCOs, and the solid précipitation was filtered off to give 1.88 g (55%) of the desired product.
Step B: 4-bromo-6,7-dihydro-5H-cyclopenta[d]pyridazin-l-amine
The mixture of 1.88 g of the product from Step A (6.8 mmol) in 25 mL of a 30% solution of 10 NH3 in water was kept at 130 °C for 18 h. After concentration, the residue was taken up in EtOAc, washed with a 10% solution of NaOH, dried on MgSO4, filtered, and concentrated to give 1.28 g (88%) of the desired product.
LC-MS-ESI (m/z): [M+H]+ calcd for C7H9BrN3: 214, 216, found: 214, 216.
Step C: N-(4-bromo-6,7-dihydro-5H-cyclopenta[d]pyridazin-l-yl)-l,3-benzothiazol-2-amine
The mixture of 214 mg of the product from Step B (1 mmol, 1 eq.), 321 mg of 2-bromo-l,3benzothiazole (1.50 mmol, 1.5 eq.), 160 mg of sodium hydride (60% suspension) (4 mmol, 4 eq.) in 25 mL of 1,4-dioxane was kept at reflux for 1 h. After cooling and quenching with methanol, the mixture was concentrated, treated with water, filtered off and dried. The solid was triturated with EtOAc and dried to give 275 mg (79%) of the desired product.
LC-MS-ESI (m/z): [M+H]+ calcd for Ci4Hi2BrN4S: 347, 349, found: 347, 349.
Step D: ethyl 2-[[l-(l,3-benzothiazol-2-ylamino)-6,7-dihydro-5H-cyclopenta[d]pyridazin-4yl]amino]thiazole-4-carboxylate
The mixture of 270 mg of the product from Step C (0.778 mmol, 1 eq.), 134 mg of ethyl 2aminothiazole-4-carboxylate (0.78 mmol, 1 eq.), 29 mg of Pd2(dba)3 (0.032 mmol, 0.04 eq.), 25 45 mg of Xantphos (0.078 mmol, 0.1 eq.), and 507 mg of Cs2CO3 (1.56 mmol, 2 eq.) in 10 mL of trifluoromethylbenzene was kept at 200 °C for 0.5 h. The reaction mixture was concentrated
209 and purified by préparative HPLC to give 97 mg (28%) of the desired product.
LC-MS-ESI (m/z): [M+H]+ calcd for C20H19N6O2S2: 439.5, found: 439.
Step E: 2-[[l-(l,3-benzothiazol-2-ylamino)-6,7-dihydro-5H-cyclopenta[d]pyridazin-4yl]amino]thiazole-4-carboxylic acid
The mixture of 90 mg of the product from Step D (0.22 mmol, 1 eq.), 18 mg LiOHxH2O (0.43 mmol, 2 eq.) of, and 13 drops of water in 5 mL of 1,4-dioxane was kept at reflux for 2 h. The reaction mixture was concentrated and purified by préparative HPLC to give 54 mg (64%) of the desired product.
HRMS-ESI (m/z): [M+H]+ calcd for C18H15N6O2S2: 411.0697, found: 411.0685.
Example 24: 6-[[4-(l,3-Benzothiazol-2-ylamino)-5,6,7,8-tetrahydrophthalazin-lyl]amino]pyridine-2-carboxylic acid
Step A: ethyl 6-[[4-(l,3-benzothiazol-2-ylamino)-5,6,7,8-tetrahydrophthalazin-lyl]amino]pyridine-2-carboxylate
The mixture of 245 mg of Example 17, Step B (0.68 mmol, 1 eq.), 113 mg of ethyl 6aminopyridine-2-carboxylate (0.68 mmol, 1 eq.), 25 mg of Pd2(dba)3 (0.027 mmol, 0.04 eq.), 40 mg of Xantphos (0.069 mmol, 0.1 eq.), and 443 mg of CS2CO3 (1.36 mmol, 2 eq.) in 10 mL of trifluoromethylbenzene was kept at 200 °C for 1.5 h. The reaction mixture was concentrated and purified by préparative HPLC to give 45 mg (15%) of the desired product.
LC-MS-ESI (m/z): [M+H]+ calcd for C23H23N6O2S: 447, found: 447.
Step B: 6-[[4-(l,3-benzothiazol-2-ylamino)-5,6,7,8-tetrahydrophthalazin-lyl]amino]pyridine-2-carboxylic acid
210
The mixture of 45 mg of the product from Step A (0.11 mmol, 1 eq.), 8.5 mg of LiOHxHzO (0.20 mmol, 2 eq.), and 10 drops of water in 10 mL of 1,4-dioxane was kept at reflux for 2 h. The reaction mixture was concentrated and purified by préparative HPLC to give 21 mg (50%) of the desired product.
HRMS-ESI (m/z): [M+H]+ calcd for C21H19N6O2S: 419.1290, found: 419.1294.
Example 25: 6-[[4-(l,3-Benzothiazol-2-ylamino)-6,7-dihydro-5Bcyclopenta [d] pyridazin-1 -yl] amino] py ridine-2-carboxylic acid
Step A: 6-[[4-(l,3-benzothiazol-2-ylamino)-6,7-dihydro-5H-cyclopenta[d]pyridazin-lyl]amino]pyridine-2-carboxylate
The mixture of 290 mg of Example 23, Step C (0.84 mmol, 1 eq.), 139 mg of ethyl 6aminopyridine-2-carboxylate (0.84 mmol, 1 eq.), 31 mg of Pd2(dba)3 (0.034 mmol, 0.04 eq.), 48 mg (0.083 mmol, 0.1 eq.) of Xantphos, and 545 mg of CS2CO3 (1.67 mmol, 2 eq.) in 10 mL of trifluoromethylbenzene was kept at 200 °C for 1.5 h. The reaction mixture was concentrated and purified by préparative HPLC to give 70 mg (19.4%) of the desired product.
LC-MS-ESI (m/z): [M+H]+ calcd for C22H21N6O2S: 433.5, found: 433 and 431 [M-H]'.
Step B: 6-[[4-(l,3-benzothiazol-2-ylamino)-6,7-dihydro-5H-cyclopenta[d]pyridazin-lyl]amino]pyridine-2-carboxylic acid
The mixture of 70 mg of the product from Step A (0.16 mmol, 1 eq.), 14 mg of LiOHxH2O (0.33 mmol, 2 eq.), and 10 drops of water in 10 mL of 1,4-dioxane was kept at reflux for 2 h. The reaction mixture was concentrated and purified by préparative HPLC to give 24 mg (31%) of the desired product.
HRMS-ESI (m/z): [M+H]+ calcd for C20H17N6O2S: 405.1134, found: 405.1123.
211
Example 26: 2-({6-[(l,3-Benzothiazol-2-yl)amino]-4,5-dimethylpyridazin-3yl}(methyl)ammo)-l,3-thiazole-4-carboxylic acid
Step A: ethyl 2-[(6-chloro-4,5-dimethylpyridazin-3-yl)(methyl)amino]-1,3-thiazole-4carboxylate
Sodium hydride (60% dispersion, 250 mg, 6.25 mmol, 1.16 eq) was added slowly to a solution of 3,6-dichloro-4,5-dimethylpyridazine (950 mg, 5.37 mmol, 1 eq) and ethyl 2-(methylamino)l,3-thiazole-4-carboxylate (1.2 g, 6.44 mmol, 1.2 eq) in 1,4-dioxane (20 mL) at 0 °C under a nitrogen atmosphère. The mixture was stirred for 1 h at ambient température, then at 60 °C for 2 h. Saturated aqueous ammonium chloride solution (75 mL) was added and the mixture extracted with ethyl acetate (150 mL). The organic extract was washed with water (2 x 75 mL), brine (75 mL), dried (magnésium sulfate) and concentrated in vacuo. Purification by column chromatography (100 g silica) eluting with ethyl acetate afforded the desired product as a brown gum (715 mg, 2.19 mmol, 41%).
LC/MS (C13H15CIN4O2S) 327 [M+H]+; RT 1.09 (LCMS-V-B1) 'H NMR (400 MHz, CDCI3) 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-[(l,3-benzothiazol-2-yl)amino]-4,5-dimethylpyridazin-3yl] (methyl) amino) -1,3-thiazole-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(dibenzylîdeneacetone)dipalladium(0) (63.1 mg, 0.07 mmol, 0.05 eq) and Xantphos (79.7 mg, 0.14 mmol, 0.1 eq) in 1,4-dioxane (20 mL) under a nitrogen atmosphère and the mixture stirred. A,A-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 h. The solution was allowed to cool to ambient température and concentrated in vacuo. Purification by flash column chromatography (50 g silica) eluting with 1% methanol in dichloromethane gave a dark yellow gum that was triturated with diethyl ether (15 mL), filtered, washed with diethyl
212 ) ether (15 mL) and dried under vacuum to afford the desired product as a pale yellow solid (365 mg, 0.83 mmol, 60%).
LC/MS (C20H20N6O2S2) 441 [M+H]+; RT 1.26 (LCMS-V-B1) XH NMR (400 MHz, DMSO-d6) δ 11.22 (br s, 1H), 7.87 (br s, 1H), 7.76 (s, 1H), 7.56 (br s, 5 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-[(l,3-benzothiazol-2-yl)amino]-4,5-dimethylpyridazin-3-yl}(methyl)amino)1,3-thiazole-4-carboxylic acid
To a solution of the product from Step B (200 mg, 0.45 mmol, 1 eq) in 1:3 water / 1,4-dioxane 10 (4 mL) was added lithium hydroxide monohydrate (95.3 mg, 2.27 mmol, 5 eq) and the mixture was stirred at ambient température for 18 h. Water (10 mL) was added and the solution filtered through celite, the solids were washed with water (5 mL) and the combined filtrate acidified with acetic acid to give a pale yellow suspension. Solids were collected by filtration, washed with water (20 mL) and dried under vacuum to afford the desired product as a pale yellow solid 15 (150 mg, 0.36 mmol, 80%).
HRMS-ESI (m/z) [M-H]- calcd for C18H15N6O2S2: 411.0703, found 411.0689.
Example 27: 2-({6-[(l,3-Benzothiazol-2-yl)amino]-5-(hydroxymethyl)-4methylpyridazin-3-yl}amino)-l,3-thiazole-4-carboxylic acid
Step A: ethyl2-({6-[(l,3-benzothiazol-2-yl)amino]-5-(methoxymethyl)-4-methylpyridazin-3yl}amino)-l,3-thiazole-4-carboxylate
To an oven-dried microwave vial was added the product from Préparation 9jb (169 mg, 0.53 mmol, 1 eq), ethyl 2-amino-l,3-thiazole-4-carboxylate (109 mg, 0.63 mmol, 1.2 eq), césium carbonate (240 mg, 0.74 mmol, 1.4 eq), Xantphos (30.5 mg, 0.05 mmol, 0.1 eq) and 25 trîs(dibenzylideneacetone)dipalladium(0) (24.1 mg, 0.03 mmol, 0.05 eq). The vessel was
213 evacuated and flushed with nitrogen (x3), and then alpha,alpha,alpha-tûRwmuAücne (10 mL) was added. The mixture was sparged with nitrogen (10 mins) then heated at 180 °C for 1 h under microwave irradiation. The mixture was concentrated in vacuo and purification by automated flash column chromatography (CombiFlash Rf, 12 g RediSep™ silica cartridge) eluting with a gradient of 0 - 5% methanol in dichloromethane afforded the desired product (138 mg, 0.30 mmol, 57%).
LC/MS (C20H20N6O3S2) 457 [M+H]+; RT 1.17 (LCMS-V-B1) *H NMR (400 MHz, DMSO-d6) 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-[(l,3-benzothiazol-2-yl)amino]-5-(hydroxymethyl)-4-methylpyridazin-3yl}amino)-l,3-thiazole-4-carboxylate
To a solution of the product from Step A (55 mg, 0.12 mmol, 1 eq) in dichloromethane (2 mL) was added boron tribromide (0.6 mL, 1 M, 0.6 mmol, 5 eq) at 0 °C and the mixture was stirred at ambient température for 1 h. The reaction was quenched with methanol and concentrated in vacuo. The solid was taken-up in methanol and refluxed for 30 min then allowed to cool to ambient température. Purification by reverse phase automated flash column chromatography (CombiFlash Rf, C18 4.3g RediSep column) eluting with a gradient of 5 95% acetonitrile afforded a crude product (53 mg) that was taken through to the next step without further purification.
Step C: 2-({6-[(l,3-benzothiazol-2-yl)amino]-5-(hydroxymethyl)-4-methylpyridazin-3yl}amino) -1,3-thiazole-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 2M aqueous lithium hydroxide (0.24 mL, 0.24 mmol, 2 eq) and the mixture was heated at reflux for 1 h. The reaction was allowed to cool to ambient température and concentrated in vacuo. The residue was taken up in minimal water, acidified to pH 6 with IM aqueous hydrochloric acid and allowed to stir for 10 mins. 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 under vacuum to afford the desired product as a brown solid (16 mg, 0.04 mmol, 32%).
HRMS-ESI (m/z) [M-H]- calcd for C17H13N6O3S2: 413.0496, found 413.0499.
214
Example 28: 2-({6-[(l,3-Benzothiazol-2-yl)amino]-5-(methoxymethyl)-4methylpyridazin-3-yl}amino)-l,3-thiazole-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 (5 mL) was added 2M aqueous lithium hydroxide (0.26 mL, 0.26 mmol, 2 eq) and the mixture was heated at reflux for 1 h. The mixture was allowed to cool to ambient température and concentrated in vacuo. The residue was taken up in minimal water, acidified to pH 6 with IM aqueous hydrochloric acid and allowed to stir for 10 mins. Solvent was removed in vacuo and the residue was triturated with 5:1 isopropanol / water. The suspension was stirred for 30 min 10 then solids were collected by filtration, washed with isopropanol and dried under vacuum to afford the desired product as an orange solid (18.5 mg, 0.04 mmol, 33%).
HRMS-ESI (m/z) [M-H]- calcd for C18H15N6O3S2: 427.0653, found 427.0650.
Example 29: 2-({6-[(l,3-Benzothiazol-2-yl)amino]-4-(methoxymethyl)-5methylpyridazin-3-yl}amino)-l,3-thiazole-4-carboxylic acid
Step A: methyl 2-({6-[(l,3-benzothiazol-2-yl)amino]-4-(methoxymethyl)-5-methylpyridazin3-yl}amino)-l,3-thiazole-4-carboxylate
To an oven-dried microwave vial was added the product from Préparation 9ja (85 mg, 0.26 mmol, 1 eq), ethyl 2-amino-l,3-thiazole-4-carboxylate (59.3 mg, 0.34 mmol, 1.3 eq), césium
215 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 flushed with nitrogen (x3), and then alpha,alpha,«//?/z«-trifluorotoluene (10 mL) was added. The mixture was sparged with nitrogen (10 mins) then heated at 180 °C for 1 h under microwave irradiation. The mixture was concentrated in vacuo and purification by automated flash column chromatography (CombiFlash Rf, 12 g RediSep™ silica cartridge) eluting with a gradient of 0 - 5% methanol in dichloromethane afforded the desired product as an orange glassy solid (82.5 mg, 0.18 mmol, 68%) .
LC/MS (C19H18N6O3S2) 443 [M+H]+; RT 1.26 (LCMS-V-B1) XH NMR (400 MHz, DMSO-d6) 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-[(l,3-benzothiazol-2-yl)amino]-4-(methoxymethyl)-5-methylpyridazin-3yl}amino) -l,3-thiazole-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 2M aqueous lithium hydroxide (0.18 mL, 0.36 mmol, 2 eq) and the mixture was heated at reflux for 1 h. The reaction was allowed to cool to ambient température and concentrated in vacuo. The residue was taken up in minimal water, acidified to pH 6 with IM aqueous hydrochloric acid and allowed to stir for 10 mins. The solvent was removed in vacuo and the residue was triturated with 5:1 isopropanol / water. The solids were collected by filtration, washed with isopropanol (2x4 mL) and dried in vacuo. Purification by reverse phase HPLC (method HPLC-V-A2) afforded the desired product as a yellow solid (22 mg, 0.05 mmol, 28%).
HRMS-ESI (m/z) [M+H]+ calcd for C18H17N6O3S2: 429.0798, found 429.0800.
Example 30: 2-({6-[(l,3-Benzothiazol-2-yI)amino]-4-(ethoxymethyl)-5-methylpyridazin3-yl}amino)-l,3-thiazole-4-carboxylic acid
216
Step A: ethyl 2-({6-[(l,3-benzothiazol-2-yl)amino]-4-(ethoxymethyl)-5-methylpyridazin-3yl}amino)-l,3-thiazole-4-carboxylate
To an oven-dried microwave vial was added the product from Préparation 9ka (113 mg, 0.34 5 mmol, 1 eq), ethyl 2-amino-l,3-thiazole-4-carboxylate (69.7 mg, 0.4 mmol, 1.2 eq), césium carbonate (154 mg, 0.47 mmol, 1.4 eq), Xantphos (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 fhished with nitrogen (x3), and then alpha,alpha,u/p/za-trifluorotoluene (7 mL) was added. The mixture was sparged with nitrogen (10 mins) then heated at 180 °C for 1 10 h under microwave irradiation. The mixture was concentrated in vacuo and purification by automated flash column chromatography (CombiFlash Rf, 24 g RediSep™ silica cartridge) eluting with a gradient of 0 — 100% ethyl acetate in zso-heptane to afford the desired product as (47 mg, 0.1 mmol, 30%).
LC/MS (C21H22N6O3S2) 471 [M+H]+; RT 1.36 (LCMS-V-B1)
Ή NMR (400 MHz, CDCI3) 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).
Ste^:2-({6-[(l,3-benzothiazol-2-yl)amino]-4-(ethoxymethyl)-5-methylpyridazin-3yl}amino)-l,3-thiazole-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 2M aqueous lithium hydroxide (0.1 mL, 0.21 mmol, 2 eq) and the mixture was heated at reflux for 2 h. The mixture was allowed to cool to ambient température and concentrated in vacuo. The residue was taken up in minimal water, acidified to pH 6 with IM aqueous hydrochloric acid and allowed to stir for 10 mins. The sample was triturated with 5:1 25 isopropanol / water and the solids collected by filtration, washed with isopropanol and dried under vacuum to afford the desired product as a brown solid (23.9 mg, 0.05 mmol, 52%).
217 φ HRMS-ESI (m/z) [M-H]- calcd for C19H17N6O3S2: 441.0809, found 441.0803.
Example 31: 2-({6-[(l,3-Benzothiazol-2-yl)amino]-5-cyclopropylpyridazin-3yl}(methyl)amino)-l,3-thiazole-4-carboxylic acid
To a solution of the product from Préparation lld, Step B (249 mg, 0.55 mmol, 1 eq) in 1,4dioxane (10 mL) was added 2M aqueous lithium hydroxide (0.55 mL, 1.1 mmol, 2 eq) and the mixture was heated at reflux for 3 h. The reaction was allowed to cool to ambient température and concentrated in vacuo. Purification by reverse phase automated flash chromatography (CombiFlash Rf, C18 43g RediSep column) eluting with a gradient of 5 - 95% acetonitrile in 10 water afforded the desired product as a yellow powder (33 mg, 0.08 mmol, 14%) [as a lithium sait].
HRMS-ESI (m/z) [M+H]+ calcd for C19H17N6O2S2: 425.0849, found 425.0851.
Example 32: 5-{l-[(Adamantan-l-yl)methyl]-5-methyl-LH-pyrazol-4-yl}-2-({6-[( 1,3beïizothiazol-2-yl)amino]pyridazin-3-yl}amino)-l,3-thiazole-4-carboxylic acid
Step A: ethyl5-{l-[(adamantan-l-yl)methyl]-5-methyl-lH-pyrazol-4-yl}-2-[(6-{[(2Z)-3-{[2(trimethylsilyl)ethoxy]methyl}-2,3-dihydro-l,3-benzothiazol-2-ylidene]amino}pyridazin-3yl)amino]-l,3-thiazole-4-carboxylate
218
To a solution of the product from Préparation llf (64 mg, 0.11 mmol, 1 eq), Préparation 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 of tetrahydrofuran / water (4 mL) was added [Ι,Γbis(diphenylphosphino)ferrocene]dichloropalladium(II) (7.71 mg, 0.01 mmol, 0.1 eq). The mixture was sparged with nitrogen (10 min) then heated at 120 °C for lh under microwave irradiation. The mixture was diluted with ethyl acetate then successively washed with water and brine. The organic extract was dried (magnésium sulfate) and the solvent removed in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 12 g RediSep™ silica cartridge) eluting with a gradient of 0 - 50% ethyl acetate in Ao-heptane afforded the desired product as a cream solid (39.2 mg, 0.05 mmol, 49%).
LC/MS (C38H48N8O3SiS2) 757 [M+H]+; RT 1.64 (LCMS-V-B1)
Step B: ethyl 5-{l-[(adamantan-1 -yl)methyl]-5-methyl-lH-pyrazol-4-yl}-2-({6-[(l,3benzothiazol-2-yl)amino]pyridazin-3-yl}amino)-l,3-thiazole-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 then trifluoroacetic acid (0.3 mL, 4.03 mmol, 78.1 eq) was added. The mixture was allowed to warm to ambient température and stirred for 18 h. The mixture was diluted with dichloromethane and washed successively with saturated aqueous sodium bicarbonate, water, and brine. The organic phase was dried (magnésium sulfate) then the solvent removed in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 12 g RediSep™ silica cartridge) eluting with a gradient of 0 - 7% methanol in dichloromethane afforded the desired product as a cream solid (10.5 mg, 0.02 mmol, 33%).
LC/MS (C32H34N8O2S2) 627 [M+H]+; RT 1.38 (LCMS-V-B1) JH NMR (400 MHz, DMSO-d6) 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-{l-[(adamantan-l-yl)methyl]-5-methyl-lH-pyrazol-4-yl}-2-({6-[(l,3-benzothiazol2-yl)amino]pyridazin-3-yl}amino)-l,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 (2 mL) was added lithium hydroxide monohydrate (3.51 mg, 0.08 mmol, 5 eq) and the mixture was heated at reflux overnighi:. The reaction was allowed to cool to ambient température and the solvent removed in vacuo. The residue was triturated with water and the solids were collected by
219
I filtration and dried under vacuum to afford the desired product as a beige solid (7.2 mg, 0.01 mmol, 71%).
HRMS-ESI (m/z) [M+H]+ calcd for C30H31N8O2S2: 599.2006, found 599.2037.
Example 33: 5-{l-[(Adamantan-l-yl)methyl]-5-methyl-LH-pyrazol-4-yl}-2-({6-[(l,35 benzothiazol-2-yl)amino]-4,5-dimethylpyridazin-3-yl}amino)-l,3-thiazole-4-carboxylic acid
Step A: ethyl5-{l-[(adamantan-1-yl)methyl]-5-methyl-lH-pyrazol-4-yl}-2-[(4,5-dimethyl-6{[(2Z)-3-{[2-(trimethylsilyl)ethoxy]methyl}-2,3-dihydro-l,3-benzothiazol-210 ylidene]amino}pyridazin-3-yl)amino]-l,3-thiazole-4-carboxylate
To a mixture of the product from Préparation lia (99 mg, 0.16 mmol, 1 eq), the product from Préparation 10a (66.6 mg, 0.19 mmol, 1.2 eq) and potassium carbonate (64.6 mg, 0 mol, 3 eq) in 3:1 tetrahydrofuran / water (4 mL) was added [l,l'-bis(diphenylphosphino)ferrocene] dichloropalladium(II) (11.4 mg, 0.02 mmol, 0.1 eq). The mixture was sparged with nitrogen 15 (10 min) then heated at 120 °C for 1 h under microwave irradiation. The reaction was diluted with ethyl acetate then successively washed with water and brine. The organic extract was dried (magnésium sulfate) then the solvent removed in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 12 g RediSep™ silica cartridge) eluting with a gradient of 0 -50% ethyl acetate in Ao-heptane afforded the desired product as a yellow glass (65.5 mg, 0.08 20 mmol, 54%).
LC/MS (C4oH52N803SiS2) 785 [M+H]+; RT 1.82 (LCMS-V-B1)
Ή NMR (400 MHz, DMSO-d6) 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), 25 0.91 (t, 2H), 0.00 (s, 9H).
220
Step B: ethyl 5-{l-[(adamantan-1 -yl)methyl]-5-methyl-lH-pyrazol-4-yl}-2-({6-[(l,3benzothiazol-2-yl)amino]-4,5-dimethylpyridazin-3-yl}amino)-l,3-thiazole-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 then trifluoroacetic acid (0.4 mL, 5.37 mmol, 64 eq) was added. The mixture was allowed to warm to ambient température and stirred for 18 h. The reaction was diluted with dichloromethane and washed successively with saturated aqueous sodium bicarbonate, water, and brine. The organics were dried (magnésium sulfate) and the solvent removed in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 12 g RediSep™ silica cartridge) eluting with a gradient of 0 - 5% methanol in dichloromethane afforded the desired product as a yellow glass (34.2 mg, 0.05 mmol, 63%).
LC/MS (C34H38N8O2S2) 655 [M+H]+; RT 1.47 (LCMS-V-B1) ‘H NMR (400 MHz, DMSO-d6) 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-{l-[(adamantan-l-yl)methyl]-5-methyl-lH-pyrazol-4-yl}-2-({6-[(l,3-benzothiazol2-yl)amino]-4,5-dïmethylpyridazin-3-yl}amino)-l,3-thiazole-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 heated at reflux for 18 h. The reaction was allowed to cool to ambient température then the solvent removed in vacuo. The residue was triturated with water, and the solids collected by filtration and dried under vacuum to afford the desired product as a yellow solid (28 mg, 0.04 mmol, 86%) [as a lithium sait].
HRMS-ESI (m/z) [M+H]+ calcd for C32H35N8O2S2: 627.2319, found 627.2357.
Example 34: 2-({6-[(l,3-BenzothiazoI-2-yl)amino]-5-(propan-2-yl)pyridazin-3yl}(methyl)amino)-l,3-thiazole-4-carboxylic acid
221
Step A: ethyl 2-{[6-chloro-5-(propan-2-yl)pyridazin-3-yl](methyl)amino}-l,3-thiazole-4carboxylate
To a solution of the product from Préparation 7c (300 mg, 1.57 mmol, 1 eq) and ethyl 25 (methylamino)-l,3-thiazole-4-carboxylate (292 mg, 1.57 mmol, 1 eq) in tetrahydrofuran (25 mL) at 0 °C was added slowly sodium hydride (60% in minerai oil; 75.4 mg, 1.88 mmol, 1.2 eq) under a nitrogen atmosphère and the mixture was stirred at 0 °C for 1 h, then at ambient température for 18 h. Saturated aqueous ammonium chloride solution (20 mL) was added and the mixture extracted with dichloromethane (75 mL). The organic extracts were washed 10 successively with water (2 x 50 mL) and brine (50 mL), then dried (magnésium sulfate) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 24 g RediSep™ silica cartridge) eluting with a gradient of 0 - 50% ethyl acetate in Aoheptane afforded the desired product as a white crystalline solid (340 mg, 1 mmol, 64%).
LC/MS (C14H17CIN4O2S) 341 [M+H]+; RT 1.31 (LCMS-V-B1) ]H NMR (400 MHz, DMSO-d6) 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).
Step B: ethyl 2-({6-[(l,3-benzothiazol-2-yl)amino]-5-(propan-2-yl)pyridazin-3yl}(methyl)amino)-l,3-thiazole-4-carboxylate
To a solution of the product from Step A (340 mg, 1 mmol, 1 eq) and 2-aminobenzothiazole 20 (180 mg, 1.2 mmol, 1.2 eq) in 1,4 dioxane (15 mL) was added Xantphos (115 mg, 0.2 mmol,
0.2 eq) and tris(dibenzylîdeneacetone)dipalladium(0) (91.4 mg, 0.1 mmol, 0.1 eq) under a nitrogen atmosphère. MN-Diisopropylethylamine (0.52 mL, 2.99 mmol, 3 eq) was added and the mixture was heated in a sealed tube at 150 °C for 18 h. The mixture was allowed to cool to ambient température and the solvent removed in vacuo. Purification by automated flash column 25 chromatography (CombiFlash Rf, 24 g RediSep™ silica cartridge) eluting with a gradient of 0 - 3% methanol in dichloromethane afforded the desired product as a yellow solid (110 mg, 0.24 mmol, 24%).
222 ! LC/MS (C21H22N6O2S2) 455 [M+H]+; RT 1.44 (LCMS-V-B1) ‘H NMR (400 MHz, DMSO-d6) 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-[(l,3-benzothiazol-2-yl)amino]-5-(propan-2-yl)pyridazin-3-yl}(methyl)amino)l,3-thiazole-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 2M aqueous lithium hydroxide (0.24 mL, 0.49 mmol, 2 eq) and the mixture was heated at reflux for 2 h. The reaction was allowed to cool to ambient température and 10 concentrated in vacuo. The crude material was triturated with diethyl ether (10 mL) and the solids collected by filtration, washed with diethyl ether and dried under vacuum. The résultant material was dissolved in water and acidified to pH 5-6 with IM aqueous hydrochloric acid then concentrated in vacuo. The solids were triturated with 4:1 isopropanol / water (5 mL), collected by filtration and dried under vacuum to afford the desired product as a yellow solid 15 (66.7 mg, 0.16 mmol, 64%).
HRMS-ESI (m/z) [M+H]+ calcd for C19H19N6O2S2: 427.1005, found 427.1042.
Example 35: 2-({6-[(l,3-Benzothiazol-2-yl)amino]-4,5-dimethylpyridazm-3-yl}amino)-5(l-{[l-(3-methoxypropyl)cyclooctyI]methyl}-5-methyl-LH-pyrazol-4-yl)-l,3-thiazole-4carboxylic acid
Step A: ethyl2-[(4,5-dimethyl-6-{[(2Z)-3-{[2-(trimethylsilyl)ethoxy]methyl}-2,3-dihydro-l,3benzothiazol-2-ylidene]amino}pyridazin-3-yl)amino]-5-(l-{[l-(3methoxypropyl)cyclooctyl]methyl}-5-methyl-lH-pyrazol-4-yl)-l,3-thiazole-4-carboxylate
223
To a mixture of the product from Préparation lia (99 mg, 0.16 mmol, 1 eq), the product from Préparation 10b (75.6 mg, 0.19 mmol, 1.2 eq) and potassium carbonate (64.6 mg, 0 mol, 3 eq) in 3:1 tetrahydrofuran / water (4 mL) was added [l,l'-bis(diphenylphosphino)ferrocene] dichloropalladium(II) (11.4 mg, 0.02 mmol, 0.1 eq). The mixture was sparged with nitrogen 5 (10 min) then heated at 120 °C for 2 h under microwave irradiation. The reaction was diluted with ethyl acetate then washed with water followed by brine. The organic extract was dried (magnésium sulfate) and the solvent removed in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 12 g RediSep™ silica cartridge) eluting with a gradient of 0 - 50% ethyl acetate in Ao-heptane afforded the desired product as an orange glass (43.4 mg, 10 0.05 mmol, 33%).
LC/MS (C42H6oN804SiS2) 834 [M+H]+; RT 1.88 (LCMS-V-B1) XH NMR (400 MHz, DMSO-d6) 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, 15 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), .07 (s, 2H), 0.91 (t, 3H), -0.11 (s, 9H).
Step B: ethyl 2-({6-[(l,3-benzothiazol-2-yl)amino]-4,5-dimethylpyridazin-3-yl}amino)-5-(l{[l-(3-methoxypropyl)cyclooctyl]methyl}-5-methyl-lH-pyrazol-4-yl)-l,3-thiazole-4carboxylate
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 température where it was stirred for 18 h. The reaction was diluted with dichloromethane then successively washed with saturated aqueous sodium bicarbonate, water, and brine. The organics were dried (magnésium sulfate) and the 25 solvent was removed in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 12 g RediSep™ silica cartridge) eluting with a gradient of 0 - 5% methanol in dichloromethane afforded the desired product as a yellow glass (21.3 mg, 0.03 mmol, 58%).
LC/MS (C36H46N8O3S2) 703 [M+H]+; RT 1.33 (LCMS-V-B1)
Ή NMR (400 MHz, DMSO-d6) δ 10.94 (br s, 1H), 7.88 (br s, 1H), 7.63 (br s, 1H), 7.57 (s, 30 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).
224 h Step C: 2-({6-[(l,3-benzothiazol-2-yl)amino]-4,5-dimethylpyridazin-3-yl}amino)-5-(l-{[l-(3methoxypropyl)cyclooctyl]methyl}-5-methyl-lH-pyrazol-4-yl)-l,3-thiazole-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 heated at reflux for 18 h. The reaction was allowed to cool to ambient température and the solvent removed in vacuo. The residue was triturated with water and solids were collected by filtration and dried under vacuum afforded the desired product as a beige solid (13.9 mg, 0.02 mmol, 68%) [as a lithium sait],
HRMS-ESI (m/z) [M+H]+ calcd for C34H43N8O3S2: 675.2894, found 675.2933.
Example 36: 2-({6-[(l,3-Benzothiazol-2-yI)amino]-4,5-dimethylpyridazin-3-yl}amino)-5( l-{[l-(3-methoxypropyl)cyclooctyl]methyl}-5-methyl-LH-pyrazol-4-yl)-l,3-thiazole-4carboxylic acid
Step A: ethyl 2-[(4,5-dimethyl-6-{[(2Z)-3-{[3-(trimethylsilyl)propoxy]methyl}-2,3-dihydro15 l,3-benzothiazol-2-ylidene]amino}pyridazin-3-yl)amino]-5-(l-{[l-(3methoxypropyl)cyclohexyl]methyl}-5-methyl-lH-pyrazol-4-yl)-l,3-thiazole-4-carboxylate
To a mixture of the product from Préparation lia (110 mg, 0.17 mmol, 1 eq), the product from Préparation 10c (81 mg, 0.22 mmol, 1.25 eq) and potassium carbonate (71.4 mg, 0.52 mmol, 3 eq) in 3:1 tetrahydrofuran / water (4 mL) was added [l,T-bis(diphenylphosphino)ferrocene] dichloropalladium(II) (12.6 mg, 0.02 mmol, 0.1 eq). The mixture was sparged with nitrogen (10 min) then heated at 120 °C for 1 h under microwave irradiation. The reaction was diluted with ethyl acetate and successively washed with water and brine. The organics were dried (magnésium sulfate) and the solvent removed in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 12 g RediSep™ silica cartridge) eluting with a gradient of 0
- 50% ethyl acetate in zso-heptane afforded the desired product as an orange gum (81.2 mg, 0.1 mmol, 59%).
225 φ LC/MS (C4oH56N804SiS2) 805 [M+H]+; RT 1.85 (LCMS-V-B1) XH NMR (400 MHz, DMSO-d6) 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, 5 8H), 1.24 - 1.14 (m, 6H), 0.91 (t, 2H), -0.11 (s, 9H).
Step B: ethyl 2-({6-[(l,3-benzothiazol-2-yl)amino]-4,5-dimethylpyridazin-3-yl}amino)-5-(l{[l-(3-methoxypropyl)cyclohexyl]methyl}-5-methyl-lH-pyrazol-4-yl)-l,3-thiazole-4carboxylate
A solution of the product from Step A (81.2 mg, 0.1 mmol, 1 eq) in dichloromethane (5 10 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 température and stirred for 18 h. The reaction was diluted with dichloromethane then successively washed with saturated aqueous sodium bicarbonate, water, and brine. The organics were dried (magnésium sulfate) and the solvent removed in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 15 12g RediSep™ silica cartridge) eluting with a gradient of 0 - 5% methanol in dichloromethane afforded the desired product as a yellow glass (49.6 mg, 0.07 mmol, 73%).
LC/MS (C34H42N8O3S2) 675 [M+H]+; RT 1.54 (LCMS-V-B1) ‘H NMR (400 MHz, DMSO-d6) δ 10.94 (br s, 1H), 7.88 (br s, 1H), 7.57 (s, 1H), 7.40 - 7.32 (m, 1H), 7.22 - 7.13 (m, 1H), 4.18 (q, 2H), 3.95 (s, 2H), 3.25 (s, 3H), 2.37 (s + s, 6H), 2.22 (s, 20 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-[(l,3-benzothiazol-2-yl)amino]-4,5-dimethylpyridazin-3-yl}amino)-5-(l-{[l-(3methoxypropyl)cyclohexyl]methyl}-5-methyl-lH-pyrazol-4-yl)-l,3-thiazole-4-carboxylic acid
To a solution of the product from Step B (49.6 mg, 0.07 mmol, 1 eq) in 1,4-dioxane was added 25 lithium hydroxide monohydrate (14.7 mg, 0.37 mmol, 5 eq) and the mixture was heated at reflux for 2 h. The reaction was allowed to cool to ambient température and the solvent removed in vacuo. The residue was triturated with water and the solids were collected by filtration, successively washed with 2M aqueous hydrochloric acid, then dichloromethane, and dried under vacuum to afford the desired product as a pale yellow solid (10.8 mg, 0.02 mmol, 23%).
HRMS-ESI (m/z) [M+H]+ calcd for C32H39N8O3S2: 647.2581, found 647.2620.
226
Example 37: 2-({6-[(l,3-Benzothiazol-2-yl)amino]-5-methylpyridazin-3yl}(ethyl)amino)-l,3-thiazole-4-carboxylic acid
Step A: ethyl2-[(6-chloro-5-methylpyridazin-3-yl)(ethyl)amino]-l,3-thiazole-4-carboxylate
Sodium hydride (60% dispersion; 150 mg, 3.75 mmol, 1.25 eq) was added to a solution of 3,6dichloro-4-methylpyridazine (530 mg, 3.25 mmol, 1.09 eq) and ethyl 2-(ethylamino)-l,3thiazole-4-carboxylate (600 mg, 3 mmol, 1 eq) in 1,4-dioxane (20 mL) at 0 °C under a nitrogen atmosphère. The mixture was stirred for 1 h at 0 °C and for 18 h at ambient température. Ethyl acetate (150 mL) was added and the mixture was successively washed with saturated aqueous 10 ammonium chloride solution (75 mL), water (2 x 75 mL), and brine (75 mL), dried (magnésium sulfate) and concentrated in vacuo. Purification by flash column chromatography (50 g silica) eluting with dichloromethane gave a pale yellow solid. Trituration with diethyl ether (10 mL) gave a solid that was collected by filtration, washed with diethyl ether (2x5 mL) and dried under vacuum to afford the desired product as an off-white solid (475 mg, 1.45 mmol, 49%).
LC/MS (C13H15CIN4O2S) 327 [M+H]+; RT 1.25 (LCMS-V-B1)
Tl NMR (400 MHz, DMSO-d6) δ 8.03 (s, 1H), 7.89 (s, 1H), 4.46 (q, 2H), 4.30 (q, 2H), 2.44 (s, 3H), 1.35 - 1.26 (m, 6H).
Step B: ethyl 2-({6-[(l,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(ethyl)amino)1,3-thiazole-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, 0.05 eq) and Xantphos (79.7 mg, 0.14 mmol, 0.1 eq) in 1,4-dioxane (10 mL) under a nitrogen atmosphère. 7V,A-Diisopropylethylamine (0.75 mL, 4.13 mmol, 3 eq) was added and the mixture was heated in a sealed tube at 150 °C for 7 h. The solution was allowed to cool to 25 ambient température and concentrated in vacuo. Purification by flash column chromatography (50 g silica) eluting with ethyl acetate gave a yellow solid. Trituration with methanol (10 mL)
227 , gave a solid that was collected by filtration, washed with methanol (2x5 mL) and dried under vacuum to afford the desired product as a yellow solid (265 mg, 0.6 mmol, 44%).
LC/MS (C20H20N6O2S2) 441 [M+H]+; RT 1.37 (LCMS-V-B1)
NMR (400 MHz, DMSO-d6) δ 11.12 (br s, 1H), 7.97 (s, 1H), 7.92 (s, 1H), 7.74 (s, 1H), 5 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-[(l,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(ethyl)amino)-l,3thiazole-4-carboxylic acid
To a solution of the product from Step B (210 mg, 0.48 mmol, 1 eq) in 1:3 water / 1,4-dioxane 10 (4 mL) was added lithium hydroxide monohydrate (80.0 mg, 1.91 mmol, 4 eq) and the mixture was stirred at ambient température overnight. Water (5 mL) was added, the mixture was filtered through celite, and the solides were washed with water (10 mL). The combined filtrate was acidified with acetic acid to give a yellow suspension and the solids were collected by filtration, washed with water (30 mL) and dried under vacuum to afford the desired product as a yellow 15 solid (170 mg, 0.41 mmol, 86%).
HRMS-ESI (m/z) [M+H]+ calcd for C18H17N6O2S2: 413.0849, found 413.0882.
Example 38: 6-({6-[(l,3-Benzothiazol-2-yl)amino]pyridazin-3yl}(methyl)amino)pyridine-2-carboxylic acid
Step A: tert-butylN-[(tert-butoxy)carbonyl]-N-(6-chloropyridazin-3-yl)carbamate
4-dimethylaminopyridine (122 mg, 1 mmol, 0.02 eq) was added to a suspension of 3-amino-6chloropyridazine (6.5 g, 50.2 mmol, 1 eq) and di-ieri-butyl dicarbonate (25.2 g, 115 mmol, 2.3 eq) in dimethylformamide (45 mL) under a nitrogen atmosphère. The suspension was heated at 50 °C for 3 h then allowed to cool to ambient température and water (350 mL) was added. The
228 h solids were collected by filtration, washed with water (250 mL) then dissolved in dichloromethane (250 mL) and the solution filtered through silica (25 g), eluting with dichloromethane. The filtrate was concentrated in vacuo and trituration with heptane (50 mL) gave a solid that was collected by filtration, washed with heptane (50 mL) and dried under 5 vacuum to afford the desired product as a white solid (9.85 g, 29.9 mmol, 60%).
LC/MS (C14H20CIN3O4) 174 [M-Boc-iBu+3H]+; RT 1.28 (LCMS-V-B1) XH NMR (400 MHz, DMSO-d6) 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) under a nitrogen atmosphère. N,NDiisopropylethylamine (5.2 mL, 29.6 mmol, 3 eq) was added and the mixture was heated at 100 °C for 3 h. The solution was allowed to cool to ambient température 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 (magnésium sulfate) and concentrated in vacuo. Purification by flash column chromatography (100 g silica) eluting with 2:3 ethyl acetate / heptane gave a yellow solid. Trituration with diethyl ether (20 mL) gave a solid that was collected by filtration, washed with diethyl ether (20 mL) and dried under vacuum to afford the desired product as a pale yellow solid (3.35 g, 7.52 mmol, 76%).
LC/MS (C21H27N5O6) 446 [M+H]+; RT 1.27 (LCMS-V-B1) XH NMR (400 MHz, DMSO-d6) δ 10.41(s, 1H), 9.02 (d, 1H), 7.97 (dd, 1H), 7.83 - 7.73 (m, 25 2H), 7.45 (d, 1H), 4.01 (s, 3H), 1.46 (s, 9H).
Step C: methyl 6-K6-{bis[(tert-butoxy) carbonyl]amino}pyridazin-3yl) (methyl) amino]pyridine-2-carboxylate
Potassium teri-butoxide (750 mg, 6.68 mmol, 1.32 eq) was added to a suspension of the product from Step B (2.25 g, 5.05 mmol, 1 eq) in dimethylsulfoxide (25 mL) under a nitrogen 30 atmosphère 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 h. The mixture was partitioned between
229 h ethyl acetate (250 mL) and saturated aqueous ammonium chloride solution (150 mL), washed with water (2 x 150 mL) and brine (100 mL), dried (magnésium sulfate) and concentrated in vacuo. Purification by flash column chromatography (100 g silica) eluting with 2:3 ethyl acetate / heptane afforded the desired product as a yellow solid (1.4 g, 2.59 mmol, 51%) that was used 5 directly in the next step without further purification.
LC/MS (C22H29N5O6) 460 [M+H]+; RT 1.29 (LCMS-V-B1).
Step D: methyl 6-[(6-aminopyridazin-3-yl)(methyl)amino]pyridine-2-carboxylate
Trifluoroacetic acid (5 mL, 64.7 mmol, 25 eq) was added to the product from Step C (1.4 g, 2.59 mmol, 1 eq) in dichloromethane (35 mL) at 0 °C and the solution was stirred for 1 h at 0 10 °C then 18 h at ambient température. The solution was concentrated in vacuo and water (25 mL) was added to give a pale yellow suspension that was neutralised with sodium bicarbonate carbonate and the solids were collected by filtration, washed with water (75 mL) and dried under vacuum to afford the desired product as a yellow solid (650 mg, 2.51 mmol, 97%) that was used directly in the next step without further purification or characterisation.
Step E: methyl 6-({6-[(l,3-benzothiazol-2-yl)amino]pyridazin-3-yl}(methyl)amino)pyridine2-carboxylate
The product from Step D (650 mg, 2.51 mmol, 1 eq) and 2-bromo-l,3-benzothiazole (600 mg, 2.8 mmol, 1.12 eq) were added to a solution of tris(dibenzylideneacetone)dipalladîum(0) (115 mg, 0.13 mmol, 0.05 eq) and Xantphos (145 mg, 0.25 mmol, 0.1 eq) in 1,4-dioxane (10 20 mL) under a nitrogen atmosphère. NjN-Diisopropylethylamine (1.4 mL, 7.52 mmol, 3 eq) was added and the mixture was heated at 100 °C for 24 h. The reaction was allowed to cool to ambient température and concentrated in vacuo. Water (20 mL) was added to give a brown suspension and the solids were collected by filtration, washed with water (40 mL) and dried under vacuum. Purificaiton by flash column chromatography (50 g silica) eluting with 39:1 25 dichloromethane / methanol gave a solid that was triturated with methanol (10 mL), filtered, washed with methanol (10 mL) and dried under vacuum to afford the desired product as a pale brown solid (225 mg, 0.57 mmol, 23%).
LC/MS (C19H16N6O2S) 393 [M+H]+; RT 1.14 (LCMS-V-B1) XH NMR (400 MHz, DMSO-d6) δ 11.73 (br s, 1H), 7.94 (d, 1H), 7.91 - 7.85 (m, 1H), 7.80 30 (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).
230
Step F: 6-({6-[(l,3-benzothiazol-2-yl)amino]pyridazin-3-yl}(methyl)amino)pyridine-2carboxylic 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 mono hydrate (85.5 mg, 2.04 mmol, 4 eq) and the mixture was stirred at ambient température for 2 h. Water (5 mL) was added, the solution was filtered through celite, and the solids washed with water (10 mL). The combined filtrate was acidified with acetic acid to give a solid, that was collected by filtration, washed with water (20 mL) and dried under vacuum to afford the desired product as a yellow solid (160 mg, 0.42 mmol, 83%).
HRMS-ESI (m/z) [M+H]+ calcd for CisHisNôChS: 379.0972, found 379.0993.
Example 39: 2-({6-[(l,3-Benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(2methoxyethyl)amiino)-l,3-thiazole-4-carboxylic acid
Step A: ethyl 2-[(6-chloro-5-methylpyridazin-3-yl)(2-methoxyethyl)amino]-l,3-thiazole-4carboxylate
Sodium hydride (60% dispersion; 650 mg, 16.3 mmol, 1.2 eq) was added to a solution of the product from Préparation 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 under a nitrogen atmosphère. The mixture was stirred for 1 h at 0 °C then heated at 60 °C for 24 h. The mixture was allowed to cool to ambient température then partitioned between ethyl acetate (300 mL) and saturated aqueous ammonium chloride solution (150 mL), and the organic phase was successively washed with water (2 x 150 mL) and brine (100 mL), dried (magnésium sulfate) and concentrated in vacuo. Trituration with diethyl ether (25 mL) gave a solid that was collected by filtration, washed with diethyl ether (15 mL) and dried under vacuum to afford the desired product as a pale pink solid (2.55 g, 7.15 mmol, 52%).
231
LC/MS (C14H17CIN4O3S) 357 [Μ+ΗΓ; RT 1.22 (LCMS-V-B1)
Ή 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).
Step B: ethyl 2-({6-[(l,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(2methoxyethyl)amino)-l,3-thiazole-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, 0.05 eq) and Xantphos (405 mg, 0.7 mmol, 0.1 eq) in 1,4-dioxane (45 mL) under a nitrogen atmosphère./V,JV-Diisopropylethylamine (3.8 mL, 21.0 mmol, 3 eq) was added and the mixture was heated in a sealed flask at 150 °C for 18 h. The solution was allowed to cool to ambient température, 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 that was collected by filtration, washed with methanol (3 x 15 mL) and dried under vacuum to afford the desired product as an orange solid (2.55 g, 5.42 mmol, 77%).
LC/MS (C21H22N6O3S2) 471 [M+H]+; RT 1.35 (LCMS-V-B1)
Ή NMR (400 MHz, CDCI3) 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-[(l,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(2methoxyethyl)amino)-1,3-thiazole-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 température for 18 h. Water (10 mL) was added and the suspension acidified with acetic acid. The solids were collected by filtration, washed with water (3 x 25 mL) and dried in vacuum to afford the desired product as a yellow solid (175 mg, 0.4 mmol, 74%).
HRMS-ESI (m/z) [M+H]+ calcd for C19H19N6O3S2: 443.0955, found 443.0983.
Example 40: 5-{l-[(Adamantan-l-yl)methyl]-5-methyl-17/-pyrazol-4-yl}-2-({6-[(l,3benzothiazol-2-yl)ammo]-5-methylpyridazin-3-yl}(methyl)amino)-l,3-thiazole-4carboxylic acid
232
Step A: ethyl5-{l-[(adamantan-1 -yl)methyl]-5-methyl-lH-pyrazol-4-yl}-2-[methyl(5methyl-6-{[(2Z)-3-{[2-(trimethylsilyl)ethoxy]methyl}-2,3-dihydro-l,3-benzothiazol-2ylidene]amino}pyridazin-3-yl)amino]-l,3-thiazole-4-carboxylate
To a mixture of the product from Préparation 11b (54.7 mg, 0.09 mmol, 1 eq), Préparation 10a (36.8 mg, 0.1 mmol, 1.2 eq) and potassium carbonate (35.7 mg, 0.26 mmol, 3 eq) in 3:1 tetrahydrofuran / water (4 mL) was added [l,l'-bis(diphenylphosphino)ferrocene] dichloropalladium(II) (6.3 mg, 0.01 mmol, 0.1 eq). The mixture was sparged with nitrogen (10 min) then heated at 120 °C for 1 h under microwave irradiation. The reaction was partitioned 10 between ethyl acetate and water, and the organic phase was washed with brine, dried (magnésium sulfate), and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 12 g RediSep™ silica cartridge) eluting with a gradient of 0 - 50% ethyl acetate in zso-heptane afforded the desired product as a brown glass (46.7 mg, 0.06 mmol, 69%).
LC/MS (C4oH52N803SiS2) 785 [M+H]+; RT 1.91 (LCMS-V-B1) XH NMR (400 MHz, DMSO-d6) δ 7.82 (dd, J = 7.7, 1.1 Hz, 1H), 7.71 (d, J = 1.1 Hz, 1H), 7.53 (s, 1H), 7.48 - 7.39 (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-{l-[(adamantan-l-yl)methyl]-5-methyl-lH-pyrazol-4-yl}-2-({6-[(l,3benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)-l,3-thiazole-4carboxylate
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 25 then the mixture was allowed to warm to ambient température and stirred for 18 h. The reaction was partitioned between dichloromethane and saturated aqueous sodium bicarbonate and the
233 organic phase was successively washed with water and brine, then dried (magnésium sulfate) and the solvent removed in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 12 g RediSep™ silica cartridge) eluting with a gradient of 0 - 5% methanol in dichloromethane afforded the desired product as a yellow glass (24.2 mg, 0.04 mmol, 62%).
LC/MS (C34H38N8O2S2) 655 [M+H]+; RT 1.53 (LCMS-V-B2) ’H NMR (400 MHz, DMSO-d6) δ 7.90 (br s, 1H), 7.73 (s, 1H), 7.54 (s, 1H), 7.37 (t, J = 7.7 Hz, 1H), 7.24 - 7.15 (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-{l-[(adamantan-1 -yl)methyl]-5-methyl-lH-pyrazol-4-yl}-2-([6-[(l,3-benzothiazol2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)-l,3-thiazole-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 heated at reflux for 18 h. The reaction was allowed to cool to ambient température then the solvent removed in vacuo. The residue was triturated with water, filtered and dried under vacuum to afford the desired product as a yellow solid (16.3 mg, 0.03 mmol, 70%) [as a lithium sait].
HRMS-ESI (m/z) [M+H]+ calcd for C32H35N8O2S2: 627.2319, found 627.2358.
Example 41: 2-({6-[(l,3-Benzothiazol-2-yl)amîno]-5-methylpyridazin-3-yl}[2(morphoIm-4-yl)ethyl]amino)-l,3-thiazole-4-carboxylic acid
Step A: ethyl2-[(2,2-dimethoxyethyl)amino]-l,3-thiazole-4-carboxylate
Ammonia (2M in isopropanol; 25 mL, 50 mmol, 2.1 eq) was added to a solution of 1,1dimethoxy-2-isothiocyanatoethane (3.5 g, 20 mmol, 1 eq) in isopropanol (15 mL) at 0 °C under a nitrogen atmosphère and the mixture stirred for 1 h at 0 °C then 18 h at ambient température. The solvent was removed in vacuo and the residue was dissolved in éthanol (60
234 mL) and cooled to 0 °C. To this was added ethyl bromopyruvate (3.6 mL, 25.8 mmol, 1.09 eq) under a nitrogen atmosphère and the mixture was allowed to warm to ambient température and stirred for 1.5 h. Triethylamine (5.6 mL, 39.9 mmol, 2 eq) was added and the mixture was stirred for 2 h then concentrated in vacuo. The residue was partitioned between ethyl acetate (250 mL) and water (150 mL) and the organic phase was successively washed with water (150 mL) and brine (150 mL), dried (magnésium sulfate), and concentrated in vacuo. Purification by flash column chromatography (100 g silica) eluting with 2:3 ethyl acetate / heptane gave a pale yellow solid that was triturated with heptane (40 mL), filtered, washed with heptane (40 mL) and dried under vacuum to afford the desired product as an off-white solid (2.95 g, 11.33 mmol, 48%).
LC/MS (C10H16N2O4S) 261 [M+H]+; RT 0.92 (LCMS-V-B1) XH NMR (400 MHz, CDCI3) 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-dimethoxy ethyl) amino]-!,3-thiazole4-carboxylate
Sodium hydride (60% dispersion; 500 mg, 12.5 mmol, 1.25 eq) 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 under a nitrogen atmosphère and the mixture was stirred for 1 h at 0 °C then for 5 h at 60 °C. The solution was allowed to cool to ambient température then partitioned between ethyl acetate (300 mL) and saturated aqueous ammonium chloride (150 mL). The organic phase was successively washed with water (2 x 150 mL) and brine (150 mL), dried (magnésium sulfate), and concentrated in vacuo. Purification by flash column chromatography (100 g silica) eluting with 99:1 dichloromethane / methanol gave an orange gum that was triturated with diethyl ether (15 mL) to give a solid that was collected by filtration, washed with diethyl ether (20 mL) and dried under vacuum to afford the desired product as a pale pink powder (1.95 g, 5.04 mmol, 50%).
LC/MS (C15H19CIN4O4S) 387 [M+H]+; RT 1.26 (LCMS-V-B1)
Ή NMR (400 MHz, DMSO-d6) 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-[(l,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(2,2dimethoxyethyl)amino)-l,3-thiazole-4-carboxylate
235
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, 0.05 eq) and Xantphos (239 mg, 0.41 mmol, 0.1 eq) in 1,4-dioxane (20 mL) under a nitrogen atmosphère and the mixture stirred. ΛζΝ-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 h. The solution was allowed to cool to ambient température and concentrated in vacuo. Trituration with diethyl ether (20 mL) gave a solid that was collected by filtration, washed with diethyl ether (2 x 10 mL) and dried under vacuum to afford the desired product as a yellow/ green solid (1.9 g, 3.42 mmol, 83%) that was used in the next step without further purification.
LC/MS (C22H24N6O4S2) 501 [M+H]+; RT 1.37 (LCMS-V-B1) 1H NMR (400 MHz, DMSO-d6) 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).
Step D: ethyl 2-({6-[(l,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 température 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 was partitioned between dichloromethane (125 mL) and 10% aqueous potassium carbona te (75 mL) and the organic phase was successively washed with water (2 x 75 mL) and brine (75 mL), dried (magnésium sulfate), and concentrated in vacuo. Purification by flash column chromatography (20 g silica) eluting with 19:1 dichloromethane / methanol gave a yellow solid that was triturated with diethyl ether (15 mL), filtered, washed with diethyl ether (15 mL), and dried under vacuum to afford the desired product as a pale orange solid (375 mg, 0.71 mmol, 48%).
LC/MS (C24H27N7O3S2) 526 [M+H]+; RT 1.08 (LCMS-V-B1) *H NMR (400 MHz, DMSO-d6) 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).
236
V Step E: 2-({6-[(l,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 température for 18 h. Water (10 mL) was added and the mixture was acidified with acetic acid. The solids were collected by filtration, washed with water and dried under vacuum to afford the desired product as a pale orange solid (215 mg, 0.43 mmol, 83%).
HRMS-ESI (m/z) [M-H]- calcd for C22H22N7O3S2: 496.1231, found 496.1232.
Example 42: 2-({6-[(l,3-Benzothiazol-2-yl)amino]-5-methyIpyridazin-310 yl}(methyl)amino)-5-(l-{[l-(3-methoxypropyl)cyclohexyl]methyl}-5-methyl-lH-pyrazol4-yl)-l,3-thiazole-4-carboxylic acid
Step A: ethyl 5-(l-{[l-(3-methoxypropyl)cyclohexyl]methyl}-5-methyl-lH-pyrazol-4-yl)-2[methyl(5-methyl-6-{[(2Z)-3-{[2-(trimethylsilyl)ethoxy]methyl}-2,3-dihydro-l,315 benzothiazol-2-ylidene]amino}pyridazin-3-yl)amino]-l,3-thiazole-4~carboxylate
To a mixture of the product from Préparation 11b (110 mg, 0.17 mmol, 1 eq), the product from Préparation 10c (78.2 mg, 0.21 mmol, 1.2 eq) and potassium carbonate (71.8 mg, 0.52 mmol, 3 eq) in 10:1 tetrahydrofuran (4 mL) / water (0.4 mL) was added [1,1'bis(diphenylphosphino)ferrocene] dichloropalladium(II) (12.7 mg, 0.02 mmol, 0.1 eq). The 20 mixture was spargcd with nitrogen (10 min) then heated at 120 °C for 1 h under microwave irradiation. The reaction was partitioned between ethyl acetate and water and the organic phase was washed with brine, dried (magnésium sulfate), and the solvent removed in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 12 g RediSep™ silica cartridge) eluting with a gradient of 0 - 50% ethyl acetate in zso-heptane afforded the 25 desired product as a yellow foam (56.1 mg, 0.07 mmol, 40%).
237
LC/MS (C40H56N8O4S1S2) 805 [M+H]+; RT 1.74 (LCMS-V-B2)
Ή NMR (400 MHz, DMSO-d6) δ 7.92 (dd, J = 7.6,1.0 Hz, 1H), 7.82 (d, J = 1.1 Hz, 1H), 7.66 (s, 1H), 7.61 - 7.47 (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-[(l,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)5-(l-{[l-(3-methoxypropyl)cyclohexyl]methyl}-5-methyl-lH-pyrazol-4-yl)-l,3-thiazole-4carboxylate
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 température where it was stirred overnight. The solvent was removed in vacuo and purification by reverse phase automated flash chromatography (CombiFlash Rf, C18 13g RediSep column) eluting with a gradient of 5 - 95% acetonitrile (pH 4; formic acid) in water water (pH 4; formic acid) afforded the desired product as a yellow glass (34.2 mg, 0.05 mmol, 73%).
LC/MS (C34H42N8O3S2) 675 [M+H]+; RT 1.47 (LCMS-V-B2) 'H NMR (400 MHz, DMSO-d6) 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-[(l,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)-5-(l{[l-(3-methoxypropyl)cyclohexyl]methyl}-5-methyl-lH-pyrazol-4-yl)-l,3-thiazole-4carboxylic 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 heated at reflux overnight. The reaction was allowed to cool to ambient température then the solvent removed in vacuo. The residue was diluted with water, acidified with 2N aqueous hydrochloric acid, and extracted with 3:1 dichloromethane / isopropanol. The organic extracts were dried (magnésium sulfate), concentrated in vacuo, and dried under vacuum afford the desired product as a cream solid (16.7 mg, 0.03 mmol, 51%).
238
HRMS-ESI (m/z) [M+H]+ calcd for C32H39N8O3S2: 647.2581, found 647.2591.
Example 43: 2-({6-[(l,3-Benzothiazol-2-yl)amino]-5-methylpyndazm-3yl}(methyl)amino)-5-(l-{[l-(3-methoxypropyl)cyclooctyl]methyl}-5-methyl-lW-pyrazol4-yl)-l,3-thiazole-4-carboxylic acid
Step A: ethyl 5-(l-{[l-(3-methoxypropyl)cyclooctyl]methyl}-5-methyl-lH-pyrazol-4-yl)-2[methyl(5-methyl-6-{[(2Z)-3-{[2-(trimethylsilyl)ethoxy]methyl}-2,3-dihydro-l,3benzothiazol-2-ylidene]amino}pyridazin-3-yl)amino]-l,3-thiazole-4-carboxylate
To a mixture of the product from Préparation 11b (106 mg, 0.17 mmol, 1 eq), the product from Préparation 10b (80.9 mg, 0.2 mmol, 1.2 eq) and potassium carbonate (69.1 mg, 0.5 mmol, 3 eq) in 3:1 tetrahydrofuran / water (4 mL) was added [l,l'-bis(diphenylphosphino)ferrocene] dichloropalladium(II) (12.2 mg, 0.02 mmol, 0.1 eq). The mixture was sparged with nitrogen (10 min) then heated at 120 °C for 1 h under microwave irradiation. The reaction was partitioned between ethyl acetate and water and the organic phase was washed with brine, dried (magnésium sulfate), and the solvent removed in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 12 g RediSep™ silica cartridge) eluting with a gradient of 0 - 50% ethyl acetate in Ao-heptane afforded the desired product as a colourless gum (81.3 mg, 0.1 mmol, 59%).
LC/MS (C42H6oNg04SiS2) 833 [M+H]+; RT 1.81 (LCMS-V-B2)
Tl NMR (400 MHz, DMSO-d6) 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).
239
Step B: ethyl 2-({6-[(l,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)5-(l~{[l-(3-methoxypropyl)cyclooctyl]methyl}-5-methyl-lH-pyrazol-4-yl)-l,3-thiazole-4carboxylate
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 température for 18 h. The solvent was removed in vacuo and purification by automated flash column chromatography (CombiFlash Rf, 12 g RediSep™ silica cartridge) eluting with a gradient of 0 - 5% methanol in dichloromethane afforded the desired product as a white solid (81.5 mg, 0.12 mmol, > 100%).
LC/MS (C36H46N8O3S2) 703 [M+H]+; RT 1.56 (LCMS-V-B2) XH NMR (400 MHz, DMSO-d6) 7.88 (br s, 1H), 7.73 (s, 1H), 7.56 (s, 1H), 7.50 (br s,l H), 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-[(l,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)-5-(l{[l-(3-methoxypropyl)cyclooctyl]methyl}-5-methyl-lH-pyrazol-4-yl)-l,3-thiazole-4carboxylic 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 heated at reflux overnight. The reaction was allowed to cool to ambient température and the solvent removed in vacuo. The residue was diluted with water, acidified with 2N aqueous hydrochloric acid, and extracted with 3:1 dichloromethane / isopropanol. The organic extracts were dried (magnésium sulfate) and concentrated under reduced pressure to afford the desired product as a cream solid (53.6 mg, 0.08 mmol, 69%).
HRMS-ESI (m/z) [M+H]+ calcd for C34H43N8O3S2: 675.2894, found 675.2890.
Example 44: 2-({6-[(l,3-Benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}[3(morpholin-4-yl)propyl]amino)-l,3-thiazole-4-carboxylic acid
240
Step A: ethyl2-{[3-(morpholin-4-yl)propyl]amino}-l,3-thiazole-4-carboxylate
Ethyl bromopyruvate (2.5 mL, 17.9 mmol, 1.18 eq) was added to a suspension of 3-(morpholin4-yl)propylthiourea (3.1 g, 15.3 mmol, 1 eq) in éthanol (50 mL) at 0 °C under a nitrogen atmosphère and the mixture was stirred for 1 h at ambient température. Triethylamine (4.5 mL, 30.5 mmol, 2 eq) was added the mixture was heated at 85 °C for 4 h. The solution was allowed to cool to ambient température and concentrated in vacuo. 10% aqueous potassium carbonate (150 mL) was added and the mixture extracted with ethyl acetate (5 x 100 mL). The combined organic extracts were washed with water (2 x 150 mL) and brine (100 mL), then dried (magnésium sulfate) and concentrated in vacuo. Purification by flash column chromatography (100 g silica) eluting with 19:1 dichloromethane / methanol (19:1) gave a solid that was triturated with heptane (50 mL), filtered, washed with heptane (50 mL) and dried under vacuum to afford the desired product as a pale brown solid (4 g, 13.4 mmol, 88%).
LC/MS (C13H21N3O3S) 300 [M+H]+; RT 0.58 (LCMS-V-B1)
Ή NMR (400 MHz, CDCI3) 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]-l,3thiazole-4-carboxylate
Sodium hydride (60% dispersion; 350 mg, 8.75 mmol, 1.31 eq) 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 under a nitrogen atmosphère and the mixture stirred for 1 h at 0 °C then for 3 h at 60 °C. The solution was allowed to cool to ambient température then partitioned between ethyl acetate (300 mL) and saturated aqueous ammonium chloride solution (100 mL). The organic phase was successively washed with water (150 mL)
241 and brine (100 mL), dried (magnésium sulfate), and concentrated in vacuo. Purification by flash column chromatography (50 g silica) eluting with ethyl acetate gave a brown oil. Trituration with heptane (15 mL) gave a solid that was collected by filtration, washed with heptane (20 mL) and dried under vacuum to afford the desired product as a pale brown solid (1.5 g, 3.52 mmol, 53%).
LC/MS (Ci8H24C]N5O3S) 426 [M+H]+; RT 0.96 (LCMS-V-B1) XH NMR (400 MHz, CDC13) 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-[(l,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}[3-(morpholin-4yl)propyl]amino)-l,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(dibenzylîdeneacetone)dipalladium(0) (96.8 mg, 0.11 mmol, 0.05 eq) and Xantphos (122 mg, 0.21 mmol, 0.1 eq) in 1,4-dioxane (20 mL) under a nitrogen atmosphère and the mixture stirred. Λζ/V-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 h. The solution was allowed to cool to ambient température and concentrated in vacuo. Purification by flash column chromatography (50 g silica) eluting with 10:1 dichloromethane / methanol gave a red/brown gum. Trituration with diethyl ether (20 mL) gave a solid that was collected by filtration, washed with diethyl ether (2 x 10 mL) and dried under vacuum to afford the desired product as a pale brown solid (875 mg, 1.62 mmol, 77%).
LC/MS (C25H29N7O3S2) 540 [M+H]+; RT 1.12 (LCMS-V-B1) XH NMR (400 MHz, DMSO-d6) 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-[(l,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}[3-(morpholin-4yl)propyl]amino)-l,3-thiazole-4-carboxylic acid
To a solution of the product from Step C (300 mg, 0.56 mmol, 1 eq) in 3:1 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 température overnight. Water (5 mL) was added and the mixture was filtered through celite and washed through with further water (10 mL). The filtrate was acidified
242 with acetic acid and stirred for 15 min, then the solids were collected by filtration, washed with water (10 mL) and drying under vacuum to afford the desired product as a yellow solid (180 mg, 0.35 mmol, 63%).
HRMS-ESI (m/z) [M-H]- calcd for C23H24N7O3S2: 510.1388, found 510.1406.
Example 45: 2-({()-[(l,3-Benzothiazol-2-yl)amino]-5-methylpyridazin-3yl}(propyl)amino)-l,3-thiazole-4-carboxyIic acid
Step A: ethyl 2-(propylamino)-l,3-thiazole-4-carboxylate
Ethyl bromopyruvate (4 mL, 28.7 mmol, 1.13 eq) was added to a solution of propylthiourea (3 10 g, 25.4 mmol, 1 eq) in éthanol (50 mL) at 0 °C under a nitrogen atmosphère and the mixture was stirred for 1 h at ambient température. Triethylamine (7 mL, 50.8 mmol, 2 eq) was added the mixture was stirred for 18 h. The reaction mixture was partitioned between ethyl acetate (300 ml.) and water (150 mL) and the organic phase was successively washed with water (150 mL) and brine (100 mL), dried (magnésium sulfate), and concentrated in vacuo. Purification by 15 flash column chromatography (50 g silica) eluting with dichloromethane afforded the desired product as a pale brown oil that solidified upon standing (3.4 g, 15.9 mmol, 63%).
LC/MS (C9H14N2O2S) 215 [M+H]+; RT 1.04 (LCMS-V-B1)
Ή NMR (400 MHz, CDCh) 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]-l,3-thiazole-4carboxylate
Sodium hydride (60% dispersion; 500 mg, 12.5 mmol, 1.25 eq) 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,
243
10.7 mmol, 1.07 eq) in 1,4-dioxane (30 mL) at 0 °C under a nitrogen atmosphère and the mixture was stirred for 1 h at 0 C then for 18 h at ambient température. 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 (2 x 150 mL) and brine (100 mL), dried (magnésium sulfate), and concentrated in vacuo. Purification by flash column chromatography (100 g silica) eluting with dichloromethane gave a solid that was triturated with heptane (30 mL), filtered, washed with heptane (15 mL) and dried under vacuum to afford the desired product as a white solid (1.3 g, 3.81 mmol, 38%).
LC/MS (C14H17CIN4O2S) 341 [M+H]+; RT 1.32 (LCMS-V-B1)
Ή NMR (400 MHz, CDCI3) δ 7.82 (s, 1H), 7.39 (d, 1H), 4.47 - 4.35 (m, 4H), 2.48 (d, J = 1.0 Hz, 3H), 1.93 - 1.78 (m, 2H), 1.42 (t, J = 7.1 Hz, 3H), 1.05 (t, J = 7.4 Hz, 3H).
Step C: ethyl 2-({6-[(l,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(propyl)amino)1,3-thiazole-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(dîbenzylideneacetone)dipalladium(0) (114 mg, 0.12 mmol, 0.05 eq) and Xantphos (144 mg, 0.25 mmol, 0.1 eq) in 1,4-dioxane (10 mL) under a nitrogen atmosphère and the mixture stirred. AJV-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 température and concentrated in vacuo. Purification by flash column chromatography (50 g silica) eluting with 39:1 dichloromethane / methanol gave a dark yellow solid that was triturated with methanol (15 mL), filtered, washed with methanol (15 mL) and dried under vacuum to afford the desired product as a yellow solid (915 mg, 2.01 mmol, 81%).
LC/MS (C21H22N6O2S2) 455 [M+H]+; RT 1.44 (LCMS-V-B1) XH NMR (400 MHz, DMSO-d6) 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).
244
I Step D: 2-({6-[(l,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(propyl)amino)-l,3thiazole-4-carboxylic acid
To a solution of the product from Step C (250 mg, 0.55 mmol, 1 eq) in 3:1 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 température for 18 h. Water (5 mL) was added and the solution was filtered through celite and washed through with water (10 mL). The filtrate was acidified with acetic acid to give a suspension, that was collected by filtration, washed with water (10 mL) and dried under vacuum to afford the desired product as a pale yellow solid (185 mg, 0.43 mmol, 79%).
HRMS-ESI (m/z) [M-H]- calcd for C19H17N6O2S2: 425.0860, found 425.0867.
Example 46: 2-({6-[(l,3-BenzothiazoI-2-yl)amino]-5-methylpyridazin-3-yl}[3(diethylamino)propyl]amino)-l,3-thiazole-4-carboxylic acid
Step A: ethyl2-{[3-(diethylamino)propyl]amino}-l,3-thiazole-4-carboxylate
Ammonia (2M in isopropanol; 60 mL, 0.12 mol, 3.45 eq) was added to a solution of 3(diethylamino)propyl isothiocyanate (6 g, 34.8 mmol, 1 eq) in isopropanol (15 mL) at 0 °C and the mixture stirred for 1 h at 0 °C and then 18 h at ambient température. The suspension was concentrated in vacuo then triturated with heptane (50 mL), filtered, and washed with heptane (2 x 25 mL). Ethanol (150 mL) and ethyl bromopyruvate (5 mL, 39.8 mmol, 1.14 eq) were added and the mixture was stirred under a nitrogen atmosphère for 1 h. Triethylamine (10 mL, 69.7 mmol, 2 eq) was added and the mixture was heated at 85 °C for 4 h. The solution was
245 allowed to cool to ambient température and concentrated in vacuo. Purification by flash column chromatography (100 g silica) eluting with 19:1 dichloromethane / methanol afforded the desired product as a dark brown gum (3.95 g, 13.8 mmol, 40%).
LC/MS (C13H23N3O2S) 286 [M+H]+; RT 0.78 (LCMS-V-B1)
Ή NMR (400 MHz, CDCI3) 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]-l,3thiazole-4-carboxylate
Sodium hydride (60% dispersion; 700 mg, 17.5 mmol, 1.26 eq) 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 under a nitrogen atmosphère and the mixture stirred for 1 h at 0 °C then for 18 h at ambient température. The reaction was partitioned between ethyl acetate (300 mL) and water (150 mL) and the organic phase was successively washed with water (150 mL) and brine (100 mL), dried (magnésium sulfate), and concentrated in vacuo. Purification by flash column chromatography (100 g silica) eluting with 19:1 dichloromethane / methanol afforded the desired product as a brown gum (2.3 g, 5.58 mmol, 40%).
LC/MS (C18H26CIN5O2S) 412 [M+H]+; RT 1.00 (LCMS-V-B1)
Ή NMR (400 MHz, CDCI3) 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,7 = 7.1 Hz, 6H).
Step C: ethyl 2-({6-[(l,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}[3(diethylamino)propyl] amino) -1,3-thiazole-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, 0.05 eq) and Xantphos (295 mg, 0.51 mmol, 0.1 eq) in 1,4-dioxane (20 mL) under a nitrogen atmosphère and the mixture stirred. Α,Α-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 h. The solution was allowed to cool to ambient température and concentrated in vacuo. Purification by flash column chromatography (100 g silica) eluting with a gradient of 5 - 10% methanol in
246 dichloromethane gave a dark yellow gum that was triturated with diethyl ether (10 mL), filtered, washed with diethyl ether (10 mL) and dried under vacuum to afford the desired product as a yellow solid (240 mg, 0.46 mmol, 9%).
LC/MS (C25H31N7O2S2) 526 [M+H]+; RT 1.18 (LCMS-V-B1)
Ή NMR (400 MHz, DMSO-d6) 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-[(l,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}[3(diethylatnino)propyl]amino)-l,3-thiazole-4-carboxylic acid
To a solution of the product from Step C (200 mg, 0.38 mmol, 1 eq) in 3:1 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 température for 18 h. Water (5 mL) was added and the mixture was filtered through celite, washing through with water (10 mL). The filtrate was acidified with acetic acid to give a solid that was collected by filtration, washed with water (10 mL) and dried under vacuum to afford the desired product as a pale yellow solid (105 mg, 0.21 mmol, 55%).
HRMS-ESI (m/z) [M-H]- calcd for C23H26N7O2S2: 496.1595, found 496.1614.
Example 47: 2-({6-[(l,3-Benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(3methoxypropyl)ammo)-l,3-thiazole-4-carboxylic acid
To a solution of the product from Préparation 11c, Step C (300 mg, 0.62 mmol, 1 eq) in 3:1 1,4-dioxane / water (4 mL) was added lithium hydroxide monohydrate (104 mg, 2.48 mmol, 4 eq) and the mixture was stirred at ambient température overnight. Water (15 mL) was added and the mixture was acidified with acetic acid to give a solid that was collected by filtration,
247 washed with water (25 mL) and dried under vacuum to afford the desired product as an orange solid (245 mg, 0.54 mmol, 87%).
HRMS-ESI (m/z) [M+H]+ calcd for C20H21N6O3S2: 457.1111, found 457.1119.
Example 48: 2-[[4,5-Dimethyl-6-[(4-methyl-l,3-benzothiazol-2-yl)amino]pyridazin-3 yl]amino]thiazole-4-carboxylic acid
Step A: N-(6-chloro-4,5-dimethyl-pyridazin-3-yl)-4-methyl-l,3-benzothiazol-2-amine
To 611 mg of 6-chloro-4,5-dimethyl-pyridazin-3-amine (3.9 mmol, 1 eq.) and 884 mg of 2bromo-4-methyl-l,3-benzothiazole (3.9 mmol, 1 eq.) in 13 mL of 1,4-dioxane was added 1.5 g 10 of sodium hydride (60w%, 37.5 mmol, 9.6 eq.) and the mixture was stirred at reflux for 1 h.
After cooling, the reaction was poured onto ice-water and acidified with 2 N HCl. The précipitation was filtered off and washed with NaHCOs solution and dried to give 810 mg (67%) of the desired product.
LC-MS-ESI (m/z): [M+H]+ calcd for C14H14CIN4S: 305, found 305.
Step B: ethyl 2-[[4,5-diniethyl-6-[(4-methyl-l,3-benzothiazol-2-yl)amino]pyridazin-3yl]amino]thiazole-4-carboxylate
The mixture of 600 mg of the product from Step A (1.96 mmol, 1 eq.), 730 mg of ethyl 2aminothiazole-4-carboxylate (4.24 mmol, 2.1 eq.), 78 mg of Pd2(dba)3 (0.085 mmol, 0.04 eq.), 124 mg oî4,5-bis(diphenylphosphino)-9,9-dimethylxanthene (0.21 mmol, 0.1 eq.) and 2.8 mL 20 of DIPEA (8 eq.) in 16 mL of 1,4-dioxane was stirred at 185°C in a microwave reactor for 1.5 h. After cooling, the volatiles were removed and the crude intermediate was purified via flash column chromatography using heptane and EtOAc as eluents to give 328 mg (38%) of the desired product.
JH NMR (500 MHz, DMSO-dô) δ ppm 11.00/10.81 (s+brs, 2H), 7.94 (s, 1H), 7.72 (brs, 1H),
248
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); 13C NMR (125 MHz, DMSO-d6) δ ppm 126.9, 122.4, 121.7, 119.4, 60.8, 18.4, 14.7; HRMS-ESI (m/z): [M+H]+ calcd for C20H20N6O2S2: 441.1167, found: 441.1165.
Step C: 2-[[4,5-ditnethyl-6-[(4-methyl-l,3-benzothiazol-2-yl)amino]pyridazin-3- yl]amino]thiazole-4-carboxylic acid
The mixture of the product from Step B, LiOHxH2O, water and 1,4-dioxane was stirred at reflux for 1 h. After cooling and concantration, 1 N HCl was added and the précipitation was filtered off, purified further by reverse phase préparative chromatography to give the desired product.
HRMS-ESI (m/z): [M+H]+ calcd for C18H17N6O2S2: 413.0854, found: 413.0855.
Example 49: 2-({6-[(l,3-BenzothiazoI-2-yl)amino]-5-methylpyridazin-3-yl}(3hydroxypropyl)amino)-l,3-thiazole-4-carboxylic acid
Step A: ethyl 2-[(6-chloro-5-methylpyridazin-3-yl)(3-hydroxypropyl)amino]-l,3-thiazole-4carboxylate
Boron tribromide (IM in dichloromethane; 16 mL, 16.2 mmol, 2 eq) was added to a solution of the product from Préparation 11c, Step B (3 g, 8.09 mmol, 1 eq) in dichloromethane (50 mL) at -78 °C under a nitrogen atmosphère. The mixture was stirred for 1 h at -78 °C then heated at 50 °C for 1 h. The suspension was allowed to cool to ambient température then partitioned between dichloromethane (250 mL) and saturated aqueous sodium hydrogen carbonate (150 mL). The organic phase was washed sequentially with water (2 x 150 mL) and brine (100 mL), dried (magnésium sulfate), and concentrated in vacuo. Purification by flash column chromatography (50 g silica) eluting with 39:1 dichloromethane / methanol gave a solid that
249 was triturated with diethyl ether (15 mL), filtered, washed with diethyl ether (20 mL) and dried under vacuum afford the desired product as an orange solid (900 mg, 2.52 mmol, 31%).
LC/MS (C14H17CIN4O3S) 357 [M+H]+; RT 1.15 (LCMS-V-B1)
Ή NMR (400 MHz, DMSO-d6) 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-[(l,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(3hydroxypropyl) am ino) -1,3-thiazole-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, 0.05 eq) and Xantphos (81.1 mg, 0.14 mmol, 0.1 eq) in 1,4-dioxane (10 mL) under a nitrogen atmosphère and the mixture stirred. A,A-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 h. The solution was allowed to cool to ambient température and concentrated in vacuo. Purification by flash column chromatography (50 g silica) eluting with 19:1 dichloromethane / methanol gave a red/orange gum that was triturated with diethyl ether (15 mL), filtered, washed with diethyl ether (2 x 10 mL) and dried under vacuum to afford the desired product as an orange solid (490 mg, 0.94 mmol, 67%).
LC/MS (C2iH22N6O3S2) 471 [M+H]+; RT 1.31 (LCMS-V-B1)
Ή NMR (400 MHz, DMSO-d6) 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-[(l,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(3hydroxypropyl)amino)-l,3-thiazole-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 1,4dioxane / water (4 mL) was added lithium hydroxide monohydrate (88.3 mg, 2.1 mmol, 4 eq) and the mixture was stirred at ambient température for 18 h. Water (15 mL) was added and the mixture was filtered through celite. The filtrate was acidified with acetic acid to give a solid that was collected by filtration, washed with water (30 mL) and dried under vacuum to afford the desired product as a yellow solid (190 mg, 0.43 mmol, 82%).
250
HRMS-ESI (m/z) [M+H]+ calcd for C19H19N6O3S2: 443.0955, found 443.0996.
Example 50: 2-[[6-(l,3-Benzothiazol-2-ylamino)-5-methyI-pyridazin-3-yl]-methylamino]-5-[3-[4-[3-(dimethylamino)prop-l-ynyl]-2-fluoro-phenoxy]propyl]thiazole-4carboxylic acid
\
Step A: ethyl5-[3-[4-[3-(dimethylamino)prop-l-ynyl]-2-fluoro-phenoxy]propyl]-2-[methyl[5-methyl-6-[(Z)-[3-(2-trimethylsilylethoxymethyl)-l,3-benzothiazol-2ylidene]amino]pyridazin-3-yl]amino]thiazole-4-carboxylate
Using Alkylation General Procedure starting from Préparation 5g and Préparation 6b as the appropriate phénol, the desired product was obtained.
Ή NMR (500 MHz, DMSO-d6) 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); 13C NMR (125 MHz, DMSO-dô) 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; HRMS-ESI (m/z): [M+H]+ calcd for C39H49FN7O4S2SK 790.3035, found 790.3023.
Step B: 2-[[6-(l,3-benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-methyl-amino]-5-[3-[4[3-(dimethylamino)prop-l-ynyl]-2-fluoro-phenoxy]propyl]thiazole-4-carboxylic acid
Using Deprotection and Hydrolysis General Procedure starting from the product from Step A as the appropriate ethyl ester, the desired product was obtained.
HRMS-ESI (m/z): [M+H]+ calcd for C31H31FN7O3S2: 632.1908, found 632.1913.
251
Example 51: 2-({6-[(l,3-Benzothiazol-2-yl)amino]-5-methyl-4-[3-(4-methylpiperazin-lyl)propyl]pyridazin-3-yl}ammo)-l,3-thÎazole-4-carboxylic acid
Step A: ethyl 2-({6-[(l,3-benzothiazol-2-yl)amino]-4-[3-(benzyloxy)propyl]-55 methylpyridazin-3-yl}amino)-l,3-thiazole-4-carboxylate
To an oven-dried microwave vial was added the product from Préparation 9ma (500 mg, 1.18 mmol, 1 eq), ethyl 2-amino-l,3-thiazole-4-carboxylate (284 mg, 1.65 mmol, 1.4 eq), césium carbonate (575 mg, 1.76 mmol, 1.5 eq), Xantphos (68.1 mg, 0.12 mmol, 0.1 eq) and tris(dîbenzylideneacetone)dipalladium(0) (53.9 mg, 0.06 mmol, 0.05 eq). The vessel was evacuated and flushed with nitrogen (x3), and then alpha,alpha,alpha-tnüuoïotoiuene (20 mL) was added. The mixture was sparged with nitrogen (10 min) then heated at 180 °C for 1 h under microwave irradiation. The reaction was concentrated in vacuo and purification by automated flash column chromatography (CombiFlash Rf, 40 g RediSep™ silica cartridge) eluting with a gradient of 0 - 70% ethyl acetate in Ao-heptane afforded the desired product as a yellow solid (336 mg, 0.6 mmol, 51%).
LC/MS (C28H28N6O3S2) 561 [M+H]+; RT 1.53 (LCMS-V-B1)
Ή NMR (400 MHz, DMSO-d6) δ 11.08 (br s, 1H), 7.95 (d, 1H), 7.88 (br s, 1H), 7.40 - 7.33 (m, 6H), 7.32 - 7.26 (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).
Step B: ethyl 2-({6-[(l,3-benzothiazol-2-yl)amino]-4-(3-hydroxypropyl)-5-methylpyridazin3-yl}amino)-l,3-thiazole-4-carboxylate
To a cooled solution of the product from Step A (330 mg, 0.59 mmol, 1 eq) in anhydrous dichloromethane (15 mL) was added boron trichloride (IM in dichloromethane; 1.18 mL, 1.18 mmol, 2 eq) dropwise and the mixture was stirred for 3 h. The reaction was cooled to 0 °C and quenched by the addition of saturated aqueous sodium bicarbonate. The mixture was diluted
252 with dichloromethane and the layers were separated. The organic phase was washed successively with water and brine, dried (magnésium sulfate), and concentrated in vacuo to afford the desired product (301 mg, 0.64 mmol, > 100%) as a mixture with the corresponding methyl ester that was used directly in the next step without further purification.
LC/MS (C21H22N6O3S2) 471 [M+H]+; RT 1.295 (LCMS-V-B1).
Step__________C: ethyl 2-{[4-(3-hydroxypropyl)-5-methyl-6-{[(2Z)-3-{[2(trimethylsilyl)ethoxy]methyl}-2,3-dihydro-l,3-benzothiazol-2-ylidene]amino}pyridazin-3yl]amino}-l,3-thiazole-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 eq) and DMAP (3.89 mg, 0.03 mmol, 0.05 eq). The mixture was stirred for 2 h at ambient température then cooled to 0 °C and quenched by the addition of aqueous sodium bicarbonate. The layers were separated and the organic phase was washed with water followed by brine, dried (magnésium sulfate), and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 24 g RediSep™ silica cartridge) eluting with a gradient of 0 - 70% ethyl acetate in Ao-heptane afforded the desired product as an orange solid (105 mg, 0.17 mmol, 27%) that was found to be a mixture with the corresponding methyl ester. The mixture was used directly in the next step without further purification.
LC/MS (C27H36N6O4SiS2) 601 [M+H]+; RT 1.59 (LCMS-V-B1)
StepD: ethyl2-{[5-methyl-4-(3-oxopropyl)-6-{[(2Z)-3-{[2-(trimethylsilyl)ethoxy]methyl}-2,3dihydro-l,3-benzothiazol-2-ylidene]amino}pyridazin-3-yl]amino}-l,3-thiazole-4-carboxylate
To an oven-dried flask was added dimethyl sulfoxide (29.6 pL, 0.42 mmol, 2.5 eq) in dichloromethane (5 mL) and the stirred solution was cooled to -78 °C. Oxalyl chloride solution (2M; 91.5 pL, 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 (5 mL) was added dropwise and the résultant mixture was stirred at -78 °C for 1 h. Triethylamine (0.14 mL, 1 mmol, 6 eq) was added and the mixture was allowed to warm to 0 °C over 1 h. The reaction was quenched with water (10 mL), diluted with sodium bicarbonate (10 mL) and extracted with dichloromethane (2 x 50 mL). The combined organic extracts were washed with brine (50 mL), dried (magnésium sulfate) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 12 g RediSep™ silica cartridge) eluting with a gradient of 0
253
- 50% ethyl acetate in zso-heptane afforded the desired product as an orange glass (68 mg, 0.11 mmol, 68%) that was found to be a mixture with the corresponding methyl ester and was used directly in the next step without further purification.
LC/MS (C27H34N6O4SiS2) 599 [M+H]+; RT 1.62 (LCMS-V-B1).
Step E: ethyl 2-({6-[(l,3-benzothiazol-2-yl)amino]-5-methyl-4-[3-(4-methylpiperazin-lyl)propyl]pyridazin-3-yl}amino)-l,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 pL, 0.23 mmol, 2 eq) followed by sodium cyanoborohydride (14.3 mg, 0.23 mmol, 2 eq) and the mixture was stirred at ambient température for 40 min. The reaction was quenched with by the addition of IM aqueous sodium hydroxide and the mixture was extracted with dichloromethane (3 x 50 mL). The combined organic extracts were washed with water followed by brine (50 mL), dried (magnésium 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 h. The mixture was diluted with dichloromethane (20 mL) and washed with IM aqueous sodium hydroxide (20 mL). The organic phase was washed with brine, dried (magnésium sulfate) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 12 g RediSep™ silica cartridge) eluting with a gradient of 0 - 20% 7N methanolic ammonia in dichloromethane afforded the desired product as an orange glass (40 mg, 0.07 mmol, 64%) that was found to be a mixture with the corresponding methyl ester and was used directly in the next step without further purification.
LC/MS (C26H32N8O2S2) 553 [M+H]+; RT 1.17 (LCMS-V-B1).
Step F: 2-({6-[(l,3-benzothiazol-2-yl)amino]-5-methyl-4-[3-(4-methylpiperazin-lyl)propyl]pyridazin-3-yl}amino)-l,3-thiazole-4-carboxylic acid
To a solution of the product of Step E (40 mg, 0.07 mmol, 1 eq) in 1,4-dioxane (5 mL) was added 2M aqueous lithium hydroxide (0.14 mL, 0.14 mmol, 2 eq) and the mixture was heated at reflux for 1 h. The reaction was concentrated in vacuo then hydrochloric acid (4M in 1,4dioxane) was added and the mixture was stirred for 1 h. The mixture was concentrated in vacuo and triturated with diethyl ether, filtered, washed with diethyl ether and dried under vacuum to afford the desired product as a dark yellow solid (14.4 mg, 0.03 mmol, 38%) [as a bishydrochloric acid sait].
HRMS-ESI (m/z) [M+H]+ calcd for C24H29N8O2S2: 525.1849, found 525.1888.
254
Example 52: 5-{l-[(Adamantan-l-yl)methyl]-5-methyl-lW-pyrazol-4-yl}-2-({6-[(l,3benzothiazol-2-yl)ammo]-5-methylpyridazin-3-yl}(3-methoxypropyl)amino)-l,3thiazole-4-carboxylic acid
N N
Step A: ethyl5-[l-[(adamantan-1 -yl)methyl]-5-methyl-lH-pyrazol-4-yl}-2-[(3methoxypropyl)(5-methyl-6-[[(2Z)-3-[[2-(trimethylsilyl)ethoxy]methyl}-2,3-dihydro-l,3benzothiazol-2-ylidene]amino}pyridazin-3-yl)amino]-l,3-thiazole-4-carboxylate
To a mixture of the product from Préparation 11c (100 mg, 0.14 mmol, 1 eq), the product from Préparation 10a (61.6 mg, 0.17 mmol, 1.2 eq) and potassium carbonate (59.8 mg, 0.43 mmol, 3 eq) in 4:1 tetrahydrofuran / water (5 mL) was added [l,T-bis(diphenylphosphino)ferrocene] dichloropalladium(II) (10.6 mg, 0.01 mmol, 0.1 eq). The mixture was sparged with nitrogen (10 min) then heated at 120 °C for 1 h under microwave irradiation. The reaction was diluted with ethyl acetate, washed with water followed by brine, dried (magnésium sulfate), and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 12 g RediSep™ silica cartridge) eluting with a gradient of 0 - 50% ethyl acetate in Aoheptane afforded the desired product as a cream foam (89.2 mg, 0.11 mmol, 73%).
LC/MS (C43H58N8O4SiS2) 843 [M+H]+; RT 1.88 (LCMS-V-B2)
Ή NMR (400 MHz, DMSO-d6) δ 7.82 (dd, J = 7.7, 1.0 Hz, 1H), 7.70 (d, J = 1.1 Hz, 1H), 7.52 (s, 1H), 7.48 - 7.40 (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).
255
StejlJL’· ethyl5-{l-[(adamantan-1 -yl)methyl]-5-methyl-lH-pyrazol-4-yl}-2-({6-[(l,3benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(3-methoxypropyl)amino)-l,3-thiazole-4carboxylate
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 température and stirred overnight. The reaction was diluted with diluted with dichloromethane, washed with saturated aqueous sodium bicarbonate, dried (magnésium sulfate), and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 12 g RediSep™ silica cartridge) eluting with a gradient of 0 -5% methanol in dichloromethane afforded the desired product as a yellow glass (63.2 mg, 0.09 mmol, 84%).
LC/MS (C37H44N8O3S2) 713 [M+H]+; RT 1.48 (LCMS-V-B1) ‘H NMR (400 MHz, DMSO-d6) 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-{l-[(adamantan-1 -yl)methyl]-5-methyl-lH-pyrazol-4-yl}-2-({6-[(l,3-benzothiazol2-yl)amino]-5-methylpyridazin-3-yl}(3-methoxypropyl)amino)-l,3-thiazole-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 heated at reflux overnight. The reaction was allowed to cool to ambient température then concentrated in vacuo. The residue was triturated with water and the solids collected by filtration, washed with water and dried under vacuum afford the desired product as a yellow solid (11.5 mg, 0.02 mmol, 19%) [as a lithium sait].
HRMS-ESI (m/z) [M+H]+ calcd for C35H41N8O3S2: 685.2738, found 685.2753.
Example 53: 5-{l-[(Adamantan-l-yl)methyl]-5-methyl-LH-pyrazol-4-yl}-2-({6-[(l,3benzothiazol-2-yl)amino]-5-cyclopropylpyridazin-3-yl}(methyl)amino)-l,3-thiazole-4carboxylic acid
256
Step A: ethyl5-{l-[(adamantan-l-yl)methyl]-5-methyl-lH-pyrazol-4-yl}-2-[(5-cyclopropyl6-{[(2Z)-3-{[2-(trimethylsilyl)ethoxy]methyl}-2,3-dihydro-l,3-benzothiazol-2ylidene]amino}pyridazin-3-yl)(methyl)amino]-l,3-thiazole-4-carboxylate
To a mixture of the product from Préparation lld (149 mg, 0.23 mmol, 1 eq), the product from Préparation 10a (96.3 mg, 0.27 mmol, 1.2 eq) and potassium carbonate (93.4 mg, 0.68 mmol, 3 eq) in 5:2 tetrahydrofuran / water (7 mL) was added [l,T-bis(diphenylphosphino)ferrocene] dichloropalladium(II) (16.5 mg, 0.02 mmol, 0.1 eq). The mixture was sparged with nitrogen (10 min) 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 (magnésium sulfate), and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 12 g RediSep™ silica cartridge) eluting with a gradient of 0 - 50% ethyl acetate in zso-heptane afforded the desired product as a white foam (130 mg, 0.16 mmol, 71%).
LC/MS (C42H54N8O3SiS2) 811 [M+H]+; RT 1.64 (LCMS-V-B1)
Ή NMR (400 MHz, DMSO-d6) 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-{l-[(adamantan-l-yl)methyl]-5-methyl-lH-pyrazol-4-yl}-2-({6-[(l,3benzothiazol-2-yl)amino]-5-cyclopropylpyridazin-3-yl}(methyl)amino)-l,3-thiazole-4carboxylate
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 mixtrure was stirred 25 for 18 h. The reaction was partitioned between dichloromethane and saturated aqueous sodium bicarbonate, washed successively with water and brine, dried (magnésium sulfate), and
257 ) concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 12 g RediSep™ silica cartridge) eluting with a gradient of 0 - 5% methanol in dichloromethane afforded the desired product as a yellow glass (104 mg, 0.15 mmol, 90%).
LC/MS (C36H40N8O2S2) 681 [M+H]+; RT 1.45 (LCMS-V-B1) XH NMR (400 MHz, DMSO-d6) δ 11.16 (br s, 1H), 7.76 (br s, 2H), 7.51 (s, 1H), 7.41 - 7.33 (m, 1H), 7.24 - 7.14 (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-{l-[(adamantan-l-yl)methyl]-5-methyl-lH-pyrazol-4-yl}-2-({6-[(l,3-benzothiazol2-yl)amino]-5-cyclopropylpyridazin-3-yl}(methyl)amino)-l,3-thiazole-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 heated at reflux overnight. The reaction was allowed to cool to ambient température and concentrated in vacuo. The résultant solid was triturated with water, filtered, and dried under vacuum to afford the desired product as a yellow solid (83.5 mg, 0.13 mmol, 84%) [as a lithium sait].
HRMS-ESI (m/z) [M+H]+ calcd for C34H37N8O2S2: 653.2475, found 653.2475.
Example 54: 2-({6-[(l,3-Benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(3methoxypropyl)amino)-5-(3-{4-[3-(dimethylamino)prop-l-yn-l-yl]-2fluorophenoxy}propyl)-l,3-thiazole-4-carboxylic acid
Step A: ethyl 5-(3-hydroxyprop-l-yn-l-yl)-2-[(3-methoxypropyl)(5-methyl-6-{[(2Z)-3-{[2(trimethylsilyl)ethoxy]methyl}-2,3-dihydro-l,3-benzothiazol-2-ylidene]amino}pyridazin-3yl)amino]-l,3-thiazole-4-carboxylate
258
Tetrakis(triphenylphosphine)palladium(0) (140.6 mg, 0.12 mmol, 0.1 eq) was added to a stirred solution of the product from Préparation 11c (844 mg, 1.22 mmol, 1 eq), propargyl alcohol (0.28 mL, 4.86 mmol, 4 eq), triethylamine (1.02 mL, 7.3 mmol, 6 eq) and copper(I) iodide (46.4 mg, 0.24 mmol, 0.2 eq) in dimethylformamide (20 mL) under a nitrogen atmosphère and the mixture was heated at 100 °C overnight. The reaction was allowed to cool to ambient température then concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 40 g RediSep™ silica cartridge) eluting with a gradient of 0 - 100% ethyl acetate in Ao-heptane afforded the desired product as a brown gum (671 mg, 1 mmol, 83%).
LC/MS (C3iH4oN605SiS2) 669 [M+H]+; RT 1.43 (LCMS-V-B1) JH NMR (400 MHz, DMSO-d6) 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, 9H).
Step B: ethyl 5-(3-hydroxypropyl)-2-[(3-methoxypropyl)(5-methyl-6-{[(2Z)-3-{[2(trimethylsilyl)ethoxy]methyl}-2,3-dihydro-l,3-benzothiazol-2-ylidene]amino}pyridazin-3yl)amino]-l,3-thiazole-4-carboxylate
A suspension of the product from Step A (671 mg, 1 mmol, 1 eq) in 1:1 ethyl acetate / methanol (30 mL) was added to a flask containing platinum (IV) oxide (0.02 g, 0.1 mmol, 0.1 eq) under a nitrogen atmosphère. The mixture was evacuated and backfilled with nitrogen (x3) then evacuated and placed under an atmosphère of hydrogen. The mixture was shaken for 3.5 h then filtered through celite (10g) and washed through with methanol. The solvent was removed in vacuo and purification by automated flash column chromatography (CombiFlash Rf, 12 g RediSep™ silica cartridge) eluting with a gradient of 0 - 60% ethyl acetate in Ao-heptane afforded the desired product as a yellow foam (431 mg, 0.64 mmol, 64%).
LC/MS (C3iH44N6O5SiS2) 673 [M+H]+; RT 1.43 (LCMS-V-B1) 1H NMR (400 MHz, DMSO-d6) 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).
259
Step C: ethyl 5-(3-iodopropyl)-2-[(3-methoxypropyl)(5-methyl-6-{[(2Z)-3-{[2(trimethylsilyl)ethoxy]methyl}-2,3-dihydro-l,3-benzothiazol-2-ylidene]amino}pyridazin-3yl)amino]-l,3-thiazole-4-carboxylate
To a stirred solution of the product from Step B (431 mg, 0.64 mmol, 1 eq) in 4:1 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 h. The reaction mixture was diluted with with ethyl acetate and successively washed with 10% aqueous sodium thiosulfate, water, and brine, dried (magnésium sulfate), and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 12 g RediSep™ silica cartridge) eluting with a gradient of 0 - 60% ethyl acetate in isoheptane afforded the desired product as a yellow gum (378 mg, 0.48 mmol, 75%).
LC/MS (C3iH43lN6O4SiS2) 783 [M+H]+; RT 1.78 (LCMS-V-B1)
NMR (400 MHz, DMSO-d6) 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 Hz, 3H), 0.92 (t, 2H), -0.11 (s, 9H).
Step D: ethyl 5-(3-{4-[3-(dimethylamino)prop-l-yn-l-yl]-2-fluorophenoxy}propyl)-2-[(3methoxypropyl)(5-methyl-6f[(2Z)-3-{[2-(trimethylsilyl)ethoxy]methyl}-2,3-dihydro-l,3benzothiazol-2-ylidene]amino}pyridazin-3-yl)amino]-l,3-thiazole-4-carboxylate
Sodium Hydride (60% in minerai oil; 38.6 mg, 0.96 mmol, 2 eq) was added to a solution of the product from Préparation 6b (103 mg, 0.53 mmol, 1.1 eq) in dimethylformamide (8 mL). After 10 min the reaction 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 température and stirred for 5 h. The reaction was quenched with water, acidified with 2N aqueous hydrochloric acid and extracted with ethyl acetate (x2). The combined organic extracts were washed with brine, dried (magnésium sulfate) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 24 g RediSep™ silica cartridge) eluting with a gradient of 0 - 10% methanol in dichloromethane afforded the desired product as a brown foam (115 mg, 0.14 mmol, 28%).
LC/MS (C42H54FN7O5S1S2) 848 [M+H]+; RT 1.43 (LCMS-V-B1) *H NMR (400 MHz, DMSO-d6) 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
260 = 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-[(l,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(3methoxypropyl)amino)-5-(3-{4-[3-(dimethylamino)prop-l-yn-l-yl]-2fluorophenoxy}propyl)-l,3-thiazole-4-carboxylate
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 h. The reaction was diluted with dichloromethane, washed with saturated aqueous sodium bicarbonate, dried (magnésium sulfate), and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 12 g RediSep™ silica cartridge) eluting with a gradient of 0 - 10% methanol in dichloromethane afforded the desired product as a yellow glass (32.2 mg, 0.04 mmol, 33%).
LC/MS (C36H40FN7O4S2) 718 [M+H]+; RT 1.32 (LCMS-V-B1) XH NMR (400 MHz, DMSO-d6) 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-[(l,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(3methoxypropyl)amino) -5-(3-{4-[3-(dimethylamino)prop-l -yn-1 -yl]-2 fluorophenoxy}propyl)-l,3-thiazole-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 heated at reflux overnight. The reaction was allowed to cool to ambient température and concentrated in vacuo. The residue was triturated with water then centrifuged and the solvent decanted. Toluene was added to the residual solid and the mixture was sonicated, centrifuged, and the solvent decanted. Drying under vacuum afforded the desired product as a yellow solid (29.6 mg, 0.04 mmol, 96%) [as a lithium sait].
HRMS-ESI (m/z) [M+H]+ calcd for C34H37FN7O4S2: 690.2327, found 690.2318.
261
Example 55: 2-[[6-[(4-Fluoro-l,3-benzothiazol-2-yl)amino]-4,5-dimethyl-pyridazin-3y 1] amino] thiazole-4-carboxylic acid
Step A: N-(6-chloro-4,5-dimethyl-pyridazin-3-yl)-4-fluoro-l,3-benzothiazol-2-amine
To 158 mg of 6-chloro-4,5-dimethyl-pyridazin-3-amine (1 mmol, 1 eq.) and 232 mg of 2bromo-4-fluoro-l,3-benzothiazole (1 mmol, 1 eq.) in 8 mL of 1,4-dioxane was added 160 mg of sodium hydride (60w%, 4 mmol, 4 eq.) and the mixture was stirred at reflux for 1 h. After cooling, the reaction was quenched with water and the mixture was extracted with EtOAc. The combined and dried organic phases were concentrated to give 311 mg (101%) of the desired 10 product.
LC-MS-ESI (m/z): [M+H]+ calcd for C13H11CIFN4S: 309, found 309.
Step B: ethyl 2-[[6-[(4-fluoro-l,3-benzothiazol-2-yl)amino]-4,5-dimethyl-pyridazin-3yl]amino]thiazole-4-carboxylate
The mixture of 300 mg of the product from Step A (0.97 mmol, 1 eq.), 167 mg of ethyl 215 aminothiazole-4-carboxylate (0.97 mmol, 1 eq.), 33 mg of Pd2(dba)3 (0.036 mmol, 0.04 eq.), 53 mg of 4,5-bis(diphenylphosphino)-9,9-dimethylxanthene (0.092 mmol, 0.1 eq.) and 0.23 mL of DIPEA (2 eq.) in 8 mL of 1,4-dioxane was stirred at 160°C in a microwave reactor for 0.75 h. After cooling, the crude product was purified by reverse phase préparative chromatography to give 110 mg (26%) of the desired product.
XH NMR (400 MHz, DMSO-rfe) 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-l,3-benzothiazol-2-yl)amino]-4,5-dimethyl-pyridazin-3yl]amino]thiazole-4-carboxylic acid
The mixture of the product from Step B, LiOHxH2O, water and 1,4-dioxane was stirred at reflux 25 for 9 h. After cooling and concentration, 1 N HCl was added and the précipitation was filtered off to give the desired product.
262
HRMS-ESI (m/z): [M+H]+ calcd for C17H14FN6O2S2: 417.0604, found: 417.0595.
Example 56: 5-({6-[(l,3-Benzothiazol-2-yl)amino]-5-methyIpyridazin-3yl}(methyl)amino)thiophene-2-carboxylic acid
Step A: tert-butylN-[(tert-butoxy)carbonyl]-N-(6-chloro-4-methylpyridazin-3-yl)carbamate
To a solution of di-iert-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 4dimethylaminopyridine (53.2 mg, 0.44 mmol, 0.05 eq) and the mixture was stirred overnight. The reaction mixture was concentrated in vacuo and purification by flash column 10 chromatography (50 g silica) eluting with 2:3 ethyl acetate / heptane gave a yellow solid.
Trituration with heptane (15 mL) gave a solid that was collected by filtration, washed with heptane (2 x 15 mL) and dried under vacuum to afford the desired product as a pale yellow solid (1.85 g, 5.38 mmol, 62%).
LC/MS (C15H22CIN3O4) 188 [M-Boc-tBu+3H]+; RT 1.28 (LCMS-V-B1) JH NMR (400 MHz, CDC13) 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-3yl)amino]thiophene-2-carboxylate
Tris(dibenzylideneacetone)dipalladium(0) (107 mg, 0.12 mmol, 0.05 eq) was added to a stirred solution of the product from Step A (800 mg, 2.33 mmol, 1 eq), methyl 5-aminothiophene-220 carboxylate (439 mg, 2.79 mmol, 1.2 eq), Aj/V-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) under a nitrogen atmosphère and the mixture was heated at 100 °C overnight. The reaction was allowed to cool to ambient température and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 24 g RediSep™ silica cartridge) eluting with a gradient of 0 25 - 10% methanol in dichloromethane gave material that was triturated in diethyl ether and the
263 solids collected by filtration to afford the desired product as a beige solid (442 mg, 0.95 mmol,
41%).
LC/MS (C2iH28N4O6S) 465 [M+H]+; RT 1.10 (LCMS-V-B1) JH NMR (400 MHz, DMSO-d6) 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 température for 5 h. The reaction was diluted with ethyl acetate then washed with water (x2) followed by brine, dried (magnésium sulfate), and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 24 g RediSep™ silica cartridge) eluting with a gradient of 0 - 100% ethyl acetate in zso-heptane afforded the desired product as a cream solid (295 mg, 0.62 mmol, 65%).
LC/MS (C22H3oN406S) 479 [M+H]+; RT 1.23 (LCMS-V-B1) JH NMR (400 MHz, DMSO-d6) 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 température and stirred for 18 h. 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 methanolic ammonia. The solvent was removed in vacuo and the résultant solid triturated with diethyl ether and filtered to afford the desired product as a cream solid (142 mg, 0.51 mmol, 71%).
LC/MS (Ci2Hi4N4O2S) 279 [M+H]+; RT 0.67 (LCMS-V-B1) *H NMR (400 MHz, DMSO-d6) 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).
264
Step E: methyl 5-({6-[(l,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3yl}(methyl)amino)thiophene-2-carboxylate
Tris(dibenzylideneaœtone)dipalladium(0) (23.4 mg, 0.03 mmol, 0.05 eq) was added to a stirred solution of the product from Step D (142 mg, 0.51 mmol, 1 eq), 2-bromo-l,3-benzothiazole (131 mg, 0.61 mmol, 1.2 eq), A,A-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) under a nitrogen atmosphère and the mixture was heated at 100 °C for 18 h. The reaction was allowed to cool to ambient température and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 12 g RediSep™ silica cartridge) eluting with a gradient of 0 - 6% methanol in dichloromethane afforded the desired product as a brown solid (158 mg, 0.38 mmol, 75%).
LC/MS (C19H17N5O2S2) 412 [M+H]+; RT 1.18 (LCMS-V-B1)
Ή NMR (400 MHz, DMSO-d6) 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-[(l,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3yl}(methyl)amino)thiophene-2-carboxylic 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 heated at reflux overnight. The reaction was allowed to cool to ambient température and the resulting suspension was centrifuged, decanted, and dried under vacuum to afford the desired product as a yellow solid (90.5 mg, 0.23 mmol, 59%).
HRMS-ESI (m/z) [M-H]- calcd for C18H14N5O2S2: 396.0594, found 396.0576.
Example 57: 2-[[6-[(5,6-Difluoro-l,3-benzothiazol-2-yl)amino]-4,5-dimethyl-pyridazin3-yl]amino]thiazole-4-carboxylic acid
265
Step A: N-(6-chloro-4,5-dimethyl-pyridazin-3-yl)-5,6-difluoro-l,3-benzothiazol-2-amine
To 158 mg of 6-chloro-4,5-dimethyl-pyridazin-3-amine (1 mmol, 1 eq.) and 250 mg of 2bromo-5,6-difluoro-l,3-benzothiazole (1 mmol, 1 eq.) in 8 mL of 1,4-dioxane was added 160 5 mg of sodium hydride (60w%, 4 mmol, 4 eq.) and the mixture was stirred at reflux for 1 h. After cooling, the reaction was quenched with water and the mixture was extracted with EtOAc. The combined and dried organic phases were concentrated to give 300 mg (92%) of the desired product.
LC-MS-ESI (m/z): [M+H]+ calcd for C13H10CIF2N4S: 327, found 327.
Step B: ethyl 2-[[6-[(5,6-difluoro-l,3-benzothiazol-2-yl)amino]-4,5-dimethyl-pyridazin-3yl]amino]thiazole-4-carboxylate
The mixture of 300 mg of the product from Step A (0.9 mmol, 1 eq.), 155 mg of ethyl 2aminothiazole-4-carboxylate (0.9 mmol, 1 eq.), 33 mg of Pd2(dba)3 (0.036 mmol, 0.04 eq.), 53 mg of 4,5-bis(diphenylphosphino)-9,9-dimethylxanthene (0.092 mmol, 0.1 eq.) and 0.23 mL of 15 DIPEA (2 eq.) in 8 mL of 1,4-dioxane was stirred at 200°C in a microwave reactor for 1 h.
After cooling, the crude product was purified by reverse phase préparative chromatography to give 110 mg (26%) of the desired product.
LC-MS-ESI (m/z): [M+H]+ calcd for C19H17F2N6O2S2: 463, found 463.
Step C: 2-[[6-[(5,6-difluoro-l,3-benzothiazol-2-yl)amino]-4,5-dimethyl-pyridazin-320 yl]amino]thiazole-4-carboxylic acid
The mixture of the product from Step B, LiOHxH2O, water and 1,4-dioxane was stirred at reflux for 9 h. After cooling, the crude product was purified by reverse phase préparative chromatography to give the desired product.
HRMS-ESI (m/z): [M+H]+ calcd for C17H13F2N6O2S2: 435.0509, found: 435.0506.
266
Example 58: 2-[[6-[(5-Fluoro-l,3-benzothiazol-2-yl)amino]-4,5-dimethyl-pyridazin-3yl]ammo]thiazole-4-carboxylic acid
Step A: N-(6-chloro-4,5-dimethyl-pyridazin-3-yl)-5~fluoro-l,3-benzothiazol-2-amine
To 158 mg of 6-chloro-4,5-dimethyl-pyridazin-3-amine (1 mmol, 1 eq.) and 348 mg of 2bromo-5-fluoro-l,3-benzothiazole (1.5 mmol, 1.5 eq.) in 20 mL of 1,4-dioxane was added 160 mg of sodium hydride (60w%, 4 mmol, 4 eq.) and the mixture was stirred at reflux for 1 h. After cooling, the reaction was quenched with 2 mL of EtOH, concentrated, and treated with 20 mL of water. The precipitated solid was filtered off to give 300 mg (97%) of the desired product.
XH NMR (500 MHz, DMSO-îA) δ ppm 7.81 (dd, 1 H), 7.29 (dd, 1 H), 7.00 (td, 1 H), 2.34 (s, 3 H), 2.29 (s, 3 H); 13C NMR (125 MHz DMSO-îA) 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]+ calcd for C13H11CIFN4S: 309.8, found: 309 and 307 [M-H]\
Step B: ethyl 2-[[6-[(5-fluoro-l,3-benzothiazol-2-yl)amino]-4,5-dimethyl-pyridazin-3yl]amino]thiazole-4-carboxylate
The mixture of 300 mg of the product from Step A (0.97 mmol, 1 eq.), 167 mg of ethyl 2aminothiazole-4-carboxylate (0.97 mmol, 1 eq.), 36 mg of Pd2(dba)3 (0.039 mmol, 0.04 eq.), 56 mg of 4,5-bis(diphenylphosphino)-9,9-dimethylxanthene (0.097 mmol, 0.1 eq.) and 0.34 mL of DIPEA (2 eq.) in 10 mL of 1,4-dioxane was stirred at 200 °C in a microwave reactor for 1 h. After cooling, the crude product was purified by reverse phase préparative chromatography to give 150 mg (35%) of the desired product.
LC-MS-ESI (m/z): [M+H]+ calcd for C19H18FN6O2S2: 445.5, found: 445 and 443 [M-H]'.
Step Ç:2-[[6-[(5-fluoro-l,3-benzothiazol-2-yl)amino]-4,5-dimethyl-pyridazin-3yl]amino]thiazole-4-carboxylic acid
The mixture of 150 mg of the product from Step B (0.338 mmol, 1 eq.), 28 mg of LiOHxHzO
267 (0.67 mmol, 2 eq.), and 10 drops of water in 10 mL of 1,4-dioxane was stirred at reflux for 3 h.
After coolmg and concentration, the crude product was purified by reverse phase préparative chromatography to give 112 mg (79%) of the desired product.
HRMS-ESI (m/z): [M+H]+ calcd for C17H14FN6O2S2: 417.0604, found: 417.0598.
Example 59: 2-({6-[(l,3-Benzothiazol-2-yI)amino]-5-methylpyridazin-3yI}(methyl)amino)-5-[3-(2-fluorophenoxy)propyl]-l,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-l,3-benzothiazol-2-ylidene]amino}pyridazin-3yl)amino]-l,3-thiazole-4-carboxylate
Sodium Hydride (60% in minerai 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 was cooled to 0 °C and a solution of the product from Préparation 5g (177 mg, 0.24 mmol, 1 eq) in dimethylformamide (3 mL) was added. The mixture was allowed to warm to ambient température and stirred for 18 h. The reaction was quenched by the addition of water, acidified with 2N aqueous hydrochloric acid and extracted with ethyl acetate (x2). The combined organic extracts were washed with brine, dried (magnésium sulfate) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 12 g RediSep™ silica cartridge) eluting with a gradient of 0 - 10% methanol in dichloromethane afforded the desired product as a beige foam (144 mg, 0.2 mmol, 83%).
LC/MS (C34H4iFN6O4SiS2) 709 [M+H]+; RT 1.62 (LCMS-V-B1)
Ή NMR (400 MHz, DMSO-d6) 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).
268
Step B: ethyl 2-({6-[(l,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)5-[3-(2-fluorophenoxy)propyl]-l,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 température overnight. The reaction was partitioned between dichloromethane and saturated aqueous sodium bicarbonate, and the organic phase was washed with brine, dried (magnésium sulfate), and concentrated in vacuo. Purification by reverse phase automated flash chromatography (CombiFlash Rf, C18 13g RediSep column) eluting with a gradient of 5 - 95% acetonitrile (pH 4; formic acid) in water (pH 4; formic acid) afforded the desired product as a yellow solid (71 mg, 0.12 mmol, 61%).
LC/MS (C28H27FN6O3S2) 579 [M+H]+; RT 1.37 (LCMS-V-B1) XH NMR (400 MHz, DMSO-d6) 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), 3.34 - 3.24 (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-[(l,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)-5-[3(2-fluorophenoxy)propyl]-l,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 heated at reflux overnight. The reaction was allowed to cool to ambient température and concentrated in vacuo. The residue was triturated in water and the solids collected by filtration. The solid was further triturated in diethyl ether, filtered, and dried under vacuum to afford the desired product as a yellow solid (49 mg, 0.09 mmol, 73%) [as a lithium sait].
HRMS-ESI (m/z) [M+H]+ calcd for C26H24FN6O3S2: 551.1330, found 551.1335.
Example 60: 2-({6-[(l,3-Benzothiazol-2-yl)amino]-5-methylpyridazin-3yl}(methyl)amino)-5-(3-methoxypropyI)-l,3-thiazole-4-carboxylic acid
269
Step A: ethyl 5-(3-methoxyprop-l-yn-1 -yl)-2-[methyl(5-methyl-6-{[(2Z)-3-{[2(trimethylsilyl)ethoxy]methyl}-2,3-dihydro-l,3-benzothiazol-2-ylidene]amino}pyridazin-3yl)amino]-l,3-thiazole-4-carboxylate
Tetrakis(triphenylphosphine)palladium(0) (36.4 mg, 0.04 mmol, 0.1 eq) was added to a stirred solution of the product from Préparation 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 copper(I) iodide (12 mg, 0.06 mmol, 0.2 eq) in dimethylformamide (5 mL) under a nitrogen atmosphère and the mixture was heated at 100 °C overnight. The reaction was allowed to cool to ambient température then concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 12 g RediSep™ silica cartridge) eluting with a gradient of 0 - 100% ethyl acetate in zso-heptane afforded the desired product as a cream solid (78 mg, 0.12 mmol, 40%).
LC/MS (C29H36N6O4SiS2) 625 [M+H]+; RT 1.56 (LCMS-V-B1) ^I NMR (400 MHz, DMSO-d6) 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-{[(2Z)-3-{[220 (trimethylsilyl)ethoxy]methyl}-2,3-dihydro-l,3-benzothiazol-2-ylidene]amino}pyridazin-3yl)amino]-l,3-thiazole-4-carboxylate
A suspension of the product from Step A (78 mg, 0.12 mmol, 1 eq) in 1:1 ethyl acetate / methanol (8 mL) was added to a flask containing platinum (IV) oxide (2.83 mg, 0.01 mmol, 0.1 eq) under a nitrogen atmosphère. The vessel was evacuated and back-filled with nitrogen (x3) 25 then evacuated and placed under an atmosphère of hydrogen and shaken overnight. The mixture was filtered through celite (2.5g), eluted with methanol, and the solvent removed in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 12 g RediSep™
270 silica cartridge) eluting with a gradient of 0 - 10% methanol in dichloromethane afforded the desired product as a white foam (56.1 mg, 0.09 mmol, 71%).
LC/MS (C29H4oN604SiS2) 629 [M+H]+; RT 1.54 (LCMS-V-B1)
Hl NMR (400 MHz, DMSO-d6) 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-[(l,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)5-(3-methoxypropyl)-l,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 température overnight. The reaction was partitioned between dichloromethane and saturated aqueous sodium bicarbonate, and the organic phase was washed with brine, dried (magnésium sulfate), and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 12 g RediSep™ silica cartridge) eluting with a gradient of 0 - 6% methanol in dichloromethane afforded the desired product as a yellow glass (40 mg, 0.08 mmol, 90%).
LC/MS (C23H26N6O3S2) 499 [M+H]+; RT 1.25 (LCMS-V-B1) >H NMR (400 MHz, DMSO-d6) 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-[(l,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)-5-(3methoxypropyl)-l,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 heated at reflux for 5 h. The reaction was allowed to cool to ambient température, concentrated in vacuo, triturated with water and the solids collected by filtration. The solid was suspended in 1,4-dioxane (4 mL) then hydrochloric acid (4M in 1,4-dioxane; 0.5 mL) was added. The suspension was stirred at ambient température for 2 h then centrifuged and the solvent removed
271 by decanting. The solid was washed with 1,4-dioxane (2x2 mL) then again centriguged and the solvent removed by decanting. The solid was finally triturated with water, filtered, and dried under vacuum to afford the desired product as an off-white solid (28.4 mg, 0.06 mmol, 75%) [as a hydrochloric acid sait].
HRMS-ESI (m/z) [M+H]+ calcd for C21H23N6O3S2: 471.1268, found 471.1271.
Example 61: 2-({6-[(l,3-BenzothiazoI-2-yl)amino]-5-methylpyridazin-3yl}(methyl)amino)-5-[3-(cyclohexyloxy)propyl]-l,3-thiazole-4-carboxylic acid
Step A: ethyl5-[3-(cyclohexyloxy)prop-l-yn-l-yl]-2-[methyl(5-methyl-6-{[(2Z)-3-{[210 (trimethylsilyl)ethoxy]methyl}-2,3-dihydro-l,3-benzothiazol-2-ylidene]amino}pyridazin-3yl)amino]-l,3-thiazole-4-carboxylate
Tetrakis(triphenylphosphïne)palladium(0) (36.4 mg, 0.04 mmol, 0.1 eq) was added to a stirred solution of the product from Préparation 11b (200 mg, 0.31 mmol, 1 eq), (prop-2-yn-lyloxy)cyclohexane (120 mg, 0.86 mmol, 2.76 eq), triethylamine (0.26 mL, 1.88 mmol, 6 15 eq) and copper(I) iodide (12 mg, 0.06 mmol, 0.2 eq) in dimethylformamide (6 mL) under a nitrogen atmosphère and the mixture was heated at 100 °C overnight. The reaction was allowed to cool to ambient température, then diluted with ethyl acetate, washed with water (x2) followed by brine, dried (magnésium sulfate) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 12 g RediSep™ silica cartridge) eluting with a 20 gradient of 0 - 70% ethyl acetate in zso-heptane afforded the desired product as a cream solid (50 mg, 0.07 mmol, 23%).
LC/MS (C34H44N6O4SiS2) 693 [M+H]+; RT 1.66 (LCMS-V-B1) *H NMR (400 MHz, DMSO-d6) 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), 25 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),
272
1.73 - 1.65 (m, 2H), 1.55 - 1.46 (m, 2H), 1.32 (t, 3H), 1.32 - 1.22 (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-l,3-benzothiazol-2-ylidene]amino}pyridazin-3yl)amino]-l,3-thiazole-4-carboxylate
A suspension of the product from Step A (50 mg, 0.07 mmol, 1 eq) in 1:1 ethyl acetate / methanol (6 mL) was added to a flask containing platinum (IV) oxide (21.6 mg, 0.01 mmol, 0.1 eq) under a nitrogen atmosphère. The vessel was evacuated and back-filled with nitrogen (x3), then evacuated and placed under an atmosphère of hydrogen and shaken for 18 h. The reaction was filtered through celite (2.5 g), eluted with ethyl acetate and methanol, and the solvent removed in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 12 g RediSep™ silica cartridge) eluting with a gradient of 0 - 60% ethyl acetate in Ao-heptane afforded the desired product as a colourless glass (42.2 mg, 0.06 mmol, 84%).
LC/MS (C34H48N6O4SiS2) 697 [M+H]+; RT 1.71 (LCMS-V-B1)
Ή NMR (400 MHz, DMSO-d6) δ 7.82 (d, J = 7.5 Hz, 1H), 7.67 (d, J = 1.1 Hz, 1H), 7.50 7.41 (m, 2H), 7.28 - 7.23 (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-[(l,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl) amino) 5-[3-(cyclohexyloxy)propyl]-l,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 température overnight. The reaction was partitioned between dichloromethane and saturated aqueous sodium bicarbonate, and the organic phase washed with brine, dried (magnésium sulfate), and concentrated in vacuo. Purification by reverse phase automated flash chromatography (CombiFlash Rf, C18 13g RediSep column) eluting with a gradient of 5 - 95% acetonitrile (pH 4; formic acid) in water (pH 4; formic acid) afforded the desired product as a yellow glass (19.6 mg, 0.03 mmol, 57%).
LC/MS (C28H34N6O3S2) 567 [M+H]+; RT 1.43 (LCMS-V-B1)
273
Ή NMR (400 MHz, DMSO-d6) 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-[(l,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)-5-[3(cyclohexyloxy)propyl]-l,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 température and concentrated in vacuo. The residue was triturated with water, filtered, and dried under vacuum to afford the desired product as a yellow solid (11 mg, 0.02 mmol, 59%) [as a lithium sait].
HRMS-ESI (m/z) [M+H]+ calcd for C26H31N6O3S2: 539.1894, found 539.1895.
Example 62: 2-({6-[(l,3-Benzothiazol-2-yl)amino]-5-cyclopropyIpyridazin-3yl}(methyl)amino)-5-(3-{4-[3-(dimethylamino)prop-l-yn-l-yl]-2-fluorophenoxy}propyl)l,3-thiazole-4-carboxylic acid
\
Step A: ethyl2-[(5-cyclopropyl-6-{[(2Z)-3-{[2-(trimethylsilyl)ethoxy]methyl}-2,3-dihydrol,3-benzothiazol-2-ylidene]amino}pyridazin-3-yl) (methyl) amino]-5-(3-hydroxyprop-l -yn-1y 1)-1,3-thiazole-4-carboxylate
Tetrakis(triphenylphosphine)palladium(0) (422 mg, 0.36 mmol, 0.1 eq) was added to a stirred solution of the product from Préparation lid (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(I) iodide (139 mg, 0.72 mmol, 0.2 eq) in dimethylformamide (50 mL) under a nitrogen atmosphère and the
274 mixture was heated at 100 °C overnight. The reaction was allowed to cool to ambient température then concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 80 g RediSep™ silica cartridge) eluting with a gradient of 0 - 100% ethyl acetate in zso-heptane afforded the desired product as a cream solid (0.91 g, 1.43 mmol, 39%).
LC/MS (C3oH36N604SiS2) 637 [M+H]+; RT 1.44 (LCMS-V-B1) *H NMR (400 MHz, DMSO-d6) 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-{[(2Z)-3-{[2-(trimethylsilyl)ethoxy]methyl}-2,3-dihydrol,3-benzothiazol-2-ylidene]amino}pyridazin-3-yl)(methyl)amino]-5-(3-hydroxypropyl)-l,3thiazole-4-carboxylate
A suspension of the product from Step A (913 mg, 1.43 mmol, 1 eq) in 1:1 ethyl acetate / methanol (40 mL) was added to a flask containing platinum (IV) oxide (32.6 mg, 0.14 mmol, 0.1 eq) under a nitrogen atmosphère. The vessel was evacuated and back-filled with nitrogen (x3) then evacuated and placed under an atmosphère of hydrogen and shaken for 3.5 h. The reaction was filtered through celite (10 g), eluted with methanol, and the solvent removed in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 12 g RediSep™ silica cartridge) eluting with a gradient of 0 - 70% ethyl acetate in zso-heptane afforded the desired product as a cream foam (762 mg, 1.19 mmol, 83%).
LC/MS (C3oH4oN604SiS2) 641 [M+H]+; RT 1.44 (LCMS-V-B1) XH NMR (400 MHz, DMSO-d6) 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-{[(2Z)-3-{[2-(trimethylsilyl)ethoxy]methyl}-2,3-dihydrol,3-benzothiazol-2-ylidene]amino}pyridazin-3-yl)(methyl)amino]-5-(3-iodopropyl)-l,3thiazole-4-carboxylate
275
To a stirred solution of the product from Step B (762 mg, 1.19 mmol, 1 eq) in 10:3 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 température overnight. The reaction was diluted with ethyl acetate, washed with 10% aqueous sodium thiosulfate solution followed by water then brine, dried (magnésium sulfate), and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 40 g RediSep™ silica cartridge) eluting with a gradient of 0 - 60% ethyl acetate in Ao-heptane afforded the desired product as a cream foam (613 mg, 0.82 mmol, 69%).
LC/MS (C3oH39lN603SiS2) 751 [M+H]+; RT 1.62 (LCMS-V-B1)
Ή NMR (400 MHz, DMSO-d6) 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, 3FI), 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, 2FI), 0.00 (s, 9H).
Step D: ethyl2-[(5-cyclopropyl-6-{[(2Z)-3-{[2-(trimethylsilyl)ethoxy]methyl}-2,3-dihydrol,3-benzothiazol-2-ylidene]amino}pyridazin-3-yl)(methyl)amino]-5-(3-{4-[3(diniethylamino)prop-l-yn-l-yl]-2-fluorophenoxy}propyl)-l,3-thiazole-4-carboxylate
Sodium Hydride (60% in minerai oil; 65.3 mg, 1.63 mmol, 2 eq) was added to a solution of the product from Préparation 6b (189 mg, 0.98 mmol, 1.2 eq) in dimethylformamide (10 mL). After 10 min the reaction 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 température and stirred for 3.5 h. The reaction was quenched by the addition of water, acidified with 2N aqueous hydrochloric acid and extracted with dichloromethane (x2). The combined organic extracts were washed with brine, dried (magnésium sulfate) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 24 g RediSep™ silica cartridge) eluting with a gradient of 0 - 10% methanol in dichloromethane gave a solid that was further purified by préparative HPLC (method HPLC-V-A1) to afford the desired product as an off-white solid (283 mg, 0.35 mmol, 43%).
LC/MS (C4iH5oFN704SiS2) 816 [M+H]+; RT 1.41 (LCMS-V-B1) XH NMR (400 MHz, DMSO-d6) 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,
276
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-[(l,3-benzothiazol-2-yl)amino]-5-cyclopropylpyridazin-3yl}(methyl)amino)-5-(3-{4-[3-(dimethylamino)prop-l-yn-l-yl]-2-fluorophenoxy}propyl)-l,3thiazole-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 température overnight. The reaction was partitioned between dichloromethane and saturated aqueous sodium bicarbonate, washed with brine, dried (magnésium sulfate), and concentrated in vacuo. Purification by reverse phase automated flash chromatography (CombiFlash Rf, C18 13g RediSep column) eluting with a gradient of 5 - 95% acetonitrile (pH 4; formic acid) in water (pH 4; formic acid) afforded the desired product as a yellow glass (56.5 mg, 0.08 mmol, 24%).
LC/MS (C35H36FN7O3S2) 686 [M+H]+; RT 1.12 (LCMS-V-B1) *H NMR (400 MHz, DMSO-d6) 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-[(l,3-benzothiazol-2-yl)amino]-5-cyclopropylpyridazin-3-yl}(methyl)amino)-5(3-{4-[3-(dimethylamino)prop-l-yn-l-yl]-2-fluorophenoxy}propyl)-l,3-thiazole-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 h. The reaction was allowed to cool to ambient température and concentrated in vacuo. The residue was triturated with water, filtered, and dried under vacuum to afford the desired product as a yellow solid (46.5 mg, 0.07 mmol, 86%) [as a lithium sait].
HRMS-ESI (m/z) [M+H]+ calcd for C33H33FN7O3S2: 658.2065, found 658.2070.
Example 63: 2-({6-[(l,3-Benzothiazol-2-yl)amino]-5-methylpyridazin-3yl}(methyl)amino)-5-(3-phenoxypropyl)-l,3-thiazole-4-carboxylic acid
277
Step A: ethyl 2-[methyl(5-methyl-6-{[(2Z)-3-{[2-(trimethylsilyl)ethoxy]methyl}-2,3-dihydrol,3-benzothiazol-2-ylidene]amino}pyridazin-3-yl)amino]-5-(3-phenoxyprop-l-yn-l-yl)-l,3thiazole-4-carboxylate
Tetrakis(triphenylphosphine)palladium(0) (85.2 mg, 0.08 mmol, 0.1 eq) was added to a stirred solution of the product from Préparation 11b (469 mg, 0.74 mmol, 1 eq), phenyl propargyl ether (0.38 mL, 2.96 mmol, 4 eq), triethylamine (0.62 mL, 4.42 mmol, 6 eq) and copper(I) iodide (28.1 mg, 0.14 mmol, 0.2 eq) in dimethylformamide (12 mL) under a nitrogen atmosphère and the mixture was heated at 100 °C overnight. The reaction was allowed to cool to ambient température and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 40 g RediSep™ silica cartridge) eluting with a gradient of 0 - 100% ethyl acetate in zso-heptane afforded the desired product as a brown foam (296 mg, 0.43 mmol, 58%).
LC/MS (C34H38N6O4SiS2) 687 [M+H]+; RT 1.57 (LCMS-V-B1)
Ή NMR (400 MHz, DMSO-d6) 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-dihydrol,3-benzothiazol-2-ylidene]amino}pyridazin-3-yl)amino]-5-(3-phenoxypropyl)-l,3-thiazole4-carboxylate
A suspension of the product from Step A (296 mg, 0.43 mmol, 1 eq) in 1:1 ethyl acetate / methanol (10 mL) was added to a flask containing platinum (IV) oxide (9.78 mg, 0.04 mmol, 25 0.1 eq) under a nitrogen atmosphère. The vessel was evacuated and back-filled with nitrogen (x3) then evacuated and placed under an atmosphère of hydrogen and shaken overnight. The reaction was filtered through celite (2.5 g), eluted with methanol, and the solvent was removed
278 in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 12 g
RediSep™ silica cartridge) eluting with a gradient of 0 - 40% ethyl acetate in Ao-heptane afforded the desired product as a brown gum (148 mg, 0.21 mmol, 50%).
LC/MS (C34H42N6O4SiS2) 691 [M+H]+; RT 1.59 (LCMS-V-B1)
Ή NMR (400 MHz, DMSO-d6) δ 7.84 (d, J = 7.7 Hz, 1H), 7.67 (d, J = 1.2 Hz, 1H), 7.51 7.40 (m, 2H), 7.33 - 7.22 (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), 3.33 - 3.22 (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-[(l,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)· 5-(3 -phenoxypropyl)-1,3-thiazole-4-carboxylate
Hydrochloric acid (4M 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 température overnight. The reaction was partitioned between ethyl acetate and saturated aqueous sodium bicarbonate, washed with brine, dried (magnésium sulfate), and concentrated in vacuo. Purification by reverse phase automated flash chromatography (CombiFlash Rf, C18 13g RediSep column) eluting with a gradient of 5 - 95% acetonitrile (pH 4; formic acid) in water (pH 4; formic acid) afforded the desired product as a yellow glass (46.7 mg, 0.08 mmol, 39%).
LC/MS (C28H28N6O3S2) 561 [M+H]+; RT 1.37 (LCMS-V-B1) XH NMR (400 MHz, DMSO-d6) 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), 3.34 - 3.24 (m, 2H), 2.47 (s, 3H), 2.21 - 1.99 (m, 2H), 1.31 (t, J = 7.1 Hz, 3H).
Step D: 2-({6-[(l,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)-5-(3phenoxypropyl)-l,3-thiazole-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 h. The reaction was allowed to cool to ambient température and concentrated in vacuo. The residue was triturated in water, filtered, and dried under vacuum to afford the desired product as a yellow solid (28.7 mg, 0.05 mmol, 65%) [as a lithium sait].
HRMS-ESI (m/z) [M-H]- calcd for C26H23N6O3S2: 531.1279, found 531.1285.
279
Example 64: 2-({6-[(l,3-Benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(3methoxypropyl)amino)-5-(3-methoxypropyl)-l,3-thiazole-4-carboxylic acid
Step A: ethyl5-(3-methoxyprop-l-yn-1 -yl)-2-[(3-methoxypropyl)(5-methy l-6-{[(2Z)-3-{[2(trimethylsilyl)ethoxy]methyl}-2,3-dihydro-l,3-benzothiazol-2-ylidene]amino}pyridazin-3yl)amino]-l,3-thiazole-4-carboxylate
Tetrakîs(triphenylphosphine)palladium(0) (83.3 mg, 0.08 mmol, 0.1 eq) was added to a stirred solution of the product from Préparation 11c (500 mg, 0.72 mmol, 1 eq), methyl propargyl ether (0.24 mL, 2.88 mmol, 4 eq), triethylamine (0.6 mL, 4.32 mmol, 6 eq) and copper(I) iodide (27.5 mg, 0.14 mmol, 0.2 eq) in dimethylformamide (10 mL) under a nitrogen atmosphère and the mixture was heated at 100 °C overnight. The reaction was allowed to cool to ambient température then concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 40 g RediSep™ silica cartridge) eluting with a gradient of 0 - 80% ethyl acetate in zso-heptane afforded the desired product as a brown gum (337 mg, 0.49 mmol, 68%).
LC/MS (C32H42N6O5SiS2) 683 [M+H]+; RT 1.57 (LCMS-V-B1) *H NMR (400 MHz, DMSO-d6) 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-{[(2Z)-3-{[2(trimethylsilyl)ethoxy]methyl}-2,3-dihydro-l,3-benzothiazol-2-ylidene]amino}pyridazin-3yl)amino]-l,3-thiazole-4-carboxylate
280
A suspension of the product from Step A (337 mg, 0.49 mmol, 1 eq) in 1:1 ethyl acetate / methanol (10 mL) was added to a flask containing platinum (IV) oxide (11.2 mg, 0.05 mmol, 0.1 eq) under a nitrogen atmosphère. The vessel was evacuated and back-filled with nitrogen (x3) then evacuated and placed under an atmosphère of hydrogen and shaken overnight. The reaction was filtered through celite (2.5 g), eluted with ethyl acetate then methanol, and the solvent removed in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 12 g RediSep™ silica cartridge) eluting with a gradient of 0 - 3% methanol in dichloromethane afforded the desired product as a yellow gum (286 mg, 0.42 mmol, 84%).
LC/MS (C32H46N6O5SiS2) 687 [M+H]+; RT 1.58 (LCMS-V-B1)
Ή NMR (400 MHz, DMSO-d6) 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-[(l,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(3methoxypropyl) amino) -5-(3-methoxypropyl) -l,3-thiazole-4-carboxylate
Hydrochloric acid (4M 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 température overnight. The reaction was partitioned between ethyl acetate and saturated aqueous sodium bicarbonate, washed with brine, dried (magnésium sulfate) and concentrated in vacuo. Purification by reverse phase automated flash chromatography (CombiFlash Rf, C18 13g RediSep column) eluting with a gradient of 5 - 95% acetonitrile (pH 4; formic acid) in water (pH 4; formic acid) afforded the desired product as a yellow foam (162 mg, 0.29 mmol, 70%).
LC/MS (C26H32N6O4S2) 557 [M+H]+; RT 1.28 (LCMS-V-B1)
Ή NMR (400 MHz, DMSO-d6) 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-[(l,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(3methoxypropyl)amino)-5-(3-methoxypropyl)-l,3-thiazole-4-carboxylic acid
281
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 heated at reflux for 7 h. The reaction was allowed to cool to ambient température apd concentrated in vacuo then dissolved in water, acidified with 2N aqueous hydrochloric acid and re-evaporated. The residue was triturated with water, filtered, and dried under vacuum to afford the desired product as a beige solid (87.8 mg, 0.17 mmol, 57%) [as a hydrochloric acid sait].
HRMS-ESI (m/z) [M+H]+ calcd for C24H29N6O4S2: 529.1686, found 529.1685.
Example 65: 2-({6-[(l,3-Benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(3methoxypropyl)ammo)-5-(3-phenoxypropyl)-l,3-thiazole-4-carboxylic acid
Step A: ethyl2-[(3-methoxypropyl)(5-methyl-6-{[(2Z)-3-{[2-(trimethylsilyl)ethoxy]methyl}2,3-dihydro-l,3-benzothiazol-2-ylidene]amino}pyridazin-3-yl)amino]-5-(3-phenoxyprop-lyn-l-yl)-l,3-thiazole-4-carboxylate
Tetrakis(trîphenylphosphine)palladium(0) (83.3 mg, 0.08 mmol, 0.1 eq) was added to a stirred solution of the product from Préparation 11c (500 mg, 0.72 mmol, 1 eq), phenyl propargyl ether (0.36 mL, 2.88 mmol, 4 eq), triethylamine (0.6 mL, 4.32 mmol, 6 eq) and copper(I) iodide (27.5 mg, 0.14 mmol, 0.2 eq) in dimethylformamide (10 mL) under a nitrogen atmosphère and the mixture was heated at 100 °C overnight. The reaction was allowed to cool to ambient température and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 24 g RediSep™ silica cartridge) eluting with a gradient of 0 - 100% ethyl acetate in zso-heptane afforded the desired product as a brown foam (356 mg, 0.48 mmol, 66%).
LC/MS (C37H44N6O5SiS2) 745 [M+H]+; RT 1.67 (LCMS-V-B1)
282
Ή NMR (400 MHz, DMSO-d6) 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}2,3-dihydro-l ,3-benzothiazol-2-ylidene]amino}pyridazin-3-yl)amino]-5-(3-phenoxypropyl)1,3-thiazole-4-carboxylate
A suspension of the product from Step A (356 mg, 0.48 mmol, 1 eq) in ethyl acetate (5 mL) was added to a flask containing platinum (IV) oxide (10.8 mg, 0.05 mmol, 0.1 eq) under a nitrogen atmosphère. The vessel was evacuated and back-filled with nitrogen (x3) then evacuated and placed under an atmosphère of hydrogen and shaken overnight. The reaction was filtered through celite (2.5 g), eluted with ethyl acetate and methanol, and the solvent was removed in vacuo and purification by automated flash column chromatography (CombiFlash Rf, 12 g RediSep™ silica cartridge) eluting with a gradient of 0 - 40% ethyl acetate in Ao-heptane afforded the desired product as a yellow gum (286 mg, 0.38 mmol, 80%).
LC/MS (C37H48N6O5SiS2) 749 [M+H]+; RT 1.66 (LCMS-V-B1) XH NMR (400 MHz, DMSO-d6) 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-[(l,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(3methoxypropyl)amino)-5-(3-phenoxypropyl)-l,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 température overnight. The reaction was partitioned between ethyl acetate and saturated aqueous sodium bicarbonate, and the organic phase washed with brine, dried (magnésium sulfate) and concentrated in vacuo. Purification by reverse phase automated flash chromatography (CombiFlash Rf, C18 13g RediSep column) eluting with a gradient of 5 -95% acetonitrile (pH 4; formic acid) in water (pH 4; formic acid) afforded the desired product as a yellow foam (145 mg, 0.23 mmol, 62%).
283
LC/MS (C31H34N6O4S2) 619 [M+H]+; RT 1.40 (LCMS-V-B1)
Ή NMR (400 MHz, DMSO-d6) 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-[(l,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(3methoxypropyl)amino)-5-(3-phenoxypropyl)-l,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 h. The reaction was allowed to cool to ambient température and concentrated in vacuo, triturated with water, filtered, and dried under vacuum to afford the desired product as a brown solid (90.6 mg, 0.15 mmol, 65%) [as a lithium sait].
HRMS-ESI (m/z) [M+H]+ calcd for C29H31N6O4S2: 591.1843, found 591.1843.
Example 66: 2-({6-[(l,3-BenzothiazoI-2-yl)amino]-5-methylpyridazin-3-yl}(3methoxypropyl)amino)-5-[3-(cyclohexyloxy)propyl]-l,3-thiazole-4-carboxylic acid
I
N.
Nx S
Step A: ethyl5-[3-(cyclohexyloxy)prop-l-yn-1 -yl]-2-[(3-methoxypropyl)(5-methyl-6-{[(2Z)3-{[2-(trimethylsilyl)ethoxy]methyl}-2,3-dihydro-l,3-benzothiazol-2ylidene]amino}pyridazin-3-yl)amino]-l,3-thiazole-4-carboxylate
Tetrakîs(triphenylphosphine)palladium(0) (83.3 mg, 0.08 mmol, 0.1 eq) was added to a stirred solution of the product from Préparation 11c (500 mg, 0.72 mmol, 1 eq), (prop-2-yn-lyloxy)cyclohexane (400 mg, 2.88 mmol, 4 eq), triethylamine (0.60 mL, 4.32 mmol, 6 eq) and copper(I) iodide (27.5 mg, 0.14 mmol, 0.2 eq) in dimethylformamide (10 mL) under a nitrogen atmosphère and the mixture was heated at 100 °C overnight. The reaction was allowed to cool
284 h to ambient température and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 40 g RediSep™ silica cartridge) eluting with a gradient of 0 - 70% ethyl acetate in Ao-heptane afforded the desired product as a brown gum (330 mg, 0.44 mmol, 61%).
LC/MS (C37H5oN605SiS2) 751 [M+H]+; RT 1.81 (LCMS-V-B1) 1H NMR (400 MHz, DMSO-d6) 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), 1.54 - 1.45 (m, 2H), 1.36 - 1.17 (m, 7H), 10 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-{[(2Z)-3-{[2(trimethylsilyl)ethoxy]methyl}-2,3-dihydro-l,3-benzothiazol-2-ylidene]amino}pyridazin-3yl)atnino]-l,3-thiazole-4-carboxylate
A suspension of the product from Step A (330 mg, 0.44 mmol, 1 eq) in 1:1 ethyl acetate / 15 methanol (10 mL) was added to a flask containing platinum (IV) oxide (9.98 mg, 0.04 mmol,
0.1 eq) under a nitrogen atmosphère. The vessel was evacuated and back-filled with nitrogen (x3) then evacuated and placed under an atmosphère of hydrogen and shaken overnight. The reaction was filtered through celite (2.5 g), eluted with ethyl acetate and methanol, and the solvent removed in vacuo. Purification by automated flash column chromatography 20 (CombiFlash Rf, 12 g RediSep™ silica cartridge) eluting with a gradient of 0 - 40% ethyl acetate in Ao-heptane afforded the desired product as a yellow gum (272 mg, 0.36 mmol, 82%).
LC/MS (C37H54N6O5SiS2) 755 [M+H]+; RT 1.79 (LCMS-V-B1)
Hl NMR (400 MHz, DMSO-d6) 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 25 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-[(l,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(3methoxypropyl)amino)-5-[3-(cyclohexyloxy)propyl]-l,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
285 was stirred at ambient température overnight. The reaction was partitioned between ethyl acetate and saturated aqueous sodium bicarbonate, and the organic phase washed with brine, dried (magnésium sulfate), and concentrated in vacuo. Purification by reverse phase automated flash chromatography (CombiFlash Rf, C18 13g RediSep column) eluting with a gradient of 5 -95% acetonitrile (pH 4; formic acid) in water (pH 4; formic acid) afforded the desired product as a yellow foam (120 mg, 0.19 mmol, 53%).
LC/MS (C31H40N6O4S2) 625 [M+H]+; RT 1.45 (LCMS-V-B1)
Ή NMR (400 MHz, DMSO-d6) 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-[(l,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(3methoxypropyl)amino)-5-[3-(cyclohexyloxy)propyl]-l,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 overnight. The reaction was allowed to cool to ambient température and concentrated in vacuo, triturated with water, filtered, and dried under vacuum to afford the desired product as a yellow solid (98 mg, 0.16 mmol, 86%) [as a lithium sait],
HRMS-ESI (m/z) [M+H]+ calcd for C29H37N6O4S2: 597.2312, found 597.2314.
Example 67: 2-({6-[(l,3-Benzothiazol-2-yl)amino]-5-methylpyridazin-3yl}(3methoxypropyl)amino)-5-[3-(2-fluorophenoxy)propyl]-l,3-thiazole-4-carboxylic acid
286
Step A: ethyl 5-(3-hydroxyprop-l-yn-l-yl)-2-[(3-methoxypropyl)(5-methyl-6-{[(2Z)-3-{[2(trimethylsilyl)ethoxy]methyl}-2,3-dihydro-l,3-benzothiazol-2-ylidene]amino}pyridazin-3yl)amino]-l,3-thiazole-4-carboxylate
Tetrakis(trîphenylphosphine)palladium(0) (83.3 mg, 0.08 mmol, 0.1 eq) was added to a stirred solution of the product from Préparation 11c (500 mg, 0.72 mmol, 1 eq), propargyl alcohol (0.16 mL, 2.88 mmol, 4 eq), triethylamine (0.60 mL, 4.32 mmol, 6 eq) and copper(I) iodide (27.5 mg, 0.14 mmol, 0.2 eq) in dimethylformamide (10 mL) under a nitrogen atmosphère and the mixture was heated at 100 °C overnight. The reaction was allowed to cool to ambient température then concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 40 g RediSep™ silica cartridge) eluting with a gradient of 0 - 100% ethyl acetate in Ao-heptane afforded the desired product as a brown solid (362 mg, 0.54 mmol, 75%).
LC/MS (C3iH4oN605SiS2) 669 [M+H]+; RT 1.45 (LCMS-V-B1)
Ή NMR (400 MHz, DMSO-d6) 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-{[(2Z)-3-{[2(trimethylsilyl)ethoxy]methyl}-2,3-dihydro-l,3-benzothiazol-2-ylidene]amino}pyridazin-3yl)amino]-l,3-thiazole-4-carboxylate
A suspension of the product from Step A (362 mg, 0.54 mmol, 1 eq) in 1:1 ethyl acetate / methanol (10 mL) was added to a flask containing platinum (IV) oxide (12.3 mg, 0.05 mmol, 0.1 eq) under a nitrogen atmosphère. The vessel was evacuated and back-filled with nitrogen (x3) then evacuated and placed under an atmosphère of hydrogen and shaken overnight. The mixture was filtered through celite (2.5 g), eluted with methanol, and the solvent removed in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 24 g RediSep™ silica cartridge) eluting with a gradient of 0 — 80% ethyl acetate in Ao-heptane afforded the desired product as a cream foam (230 mg, 0.34 mmol, 63%).
LC/MS (C3iH44N6O5SiS2) 673 [M+H]+; RT 1.47 (LCMS-V-B1)
Ή NMR (400 MHz, DMSO-d6) 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
287
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: ethyl5-(3-iodopropyl)-2-[(3-methoxypropyl)(5-methyl-6-{[(2Z)-3-{[2(trimethylsilyl)ethoxy]methyl}-2,3-dihydro-l,3-benzothiazol-2-ylidene]amino}pyridazin-3yl)amino]-l,3-thiazole-4-carboxylate
To a stirred solution of the product from Step B (230 mg, 0.34 mmol, 1 eq) in 3:1 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 température overnight. The reaction was diluted with ethyl acetate, washed with 10% aqueous sodium thiosulfate solution followed by water then brine, dried (magnésium sulfate), and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 12 g RediSep™ silica cartridge) eluting with a gradient of 0 - 50% ethyl acetate in zso-heptane afforded the desired product as a yellow gum (129 mg, 0.16 mmol, 48%).
LC/MS (C3iH43IN6O4SiS2) 783 [M+H]+; RT 1.63 (LCMS-V-B1) 'FI NMR (400 MHz, DMSO-d6) 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-{[(2Z)-3-{[2(trimethylsilyl)ethoxy]methyl}-2,3-dihydro-l,3-benzothiazol-2-ylidene]amino}pyridazin-3yl)amino]-l,3-thiazole-4-carboxylate
Sodium Hydride (60% in minerai 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 température and stirred overnight. The reaction was quenched by the addition of water, acidified with 2N aqueous hydrochloric acid, and extracted with ethyl acetate (x2). The combined organic extracts were washed with brine, dried (magnésium sulfate) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 12 g RediSep™
288 silica cartridge) eluting with a gradient of 0 - 6% methanol in dichloromethane afforded the desired product as a cream gum (75.6 mg, 0.1 mmol, 60%).
LC/MS (C37H47FN6O5SiS2) 767 [M+H]+; RT 1.61 (LCMS-V-B1)
Ή NMR (400 MHz, DMSO-d6) δ 7.84 (d, 1H), 7.67 (s, 1H), 7.51 - 7.40 (m, 2H), 7.30 - 7.07 (m, 4H), 6.97 - 6.89 (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-[(l,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(3methoxypropyl)amino)-5-[3-(2-fluorophenoxy)propyl]-l,3-thiazole-4-carboxylate
Hydrochloric acid (4M 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 température overnight. The reaction was partitioned between ethyl acetate and saturated aqueous sodium bicarbonate and the organic phase was washed with brine, dried (magnésium sulfate), and concentrated in vacuo. Purification by reverse phase automated flash chromatography (CombiFlash Rf, C18 13g RediSep column) eluting with a gradient of 5 -95% acetonitrile (pH 4; formic acid) in water (pH 4; formic acid) afforded the desired product as a yellow glass (36.2 mg, 0.06 mmol, 58%).
LC/MS (C3iH33FN6O4S2) 637 [M+H]+; RT 1.39 (LCMS-V-B1) XH NMR (400 MHz, DMSO-d6) 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-[(l,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(3methoxypropyl)amino)-5-[3-(2-fluorophenoxy)propyl]-l,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 heated at reflux overnight. The reaction was allowed to cool to ambient température and concentrated in vacuo. Purification by reverse phase automated flash chromatography (CombiFlash Rf, C18 13g RediSep column) eluting with a gradient of 5 - 95% acetonitrile (pH 4; formic acid) in
289 water (pH 4; formic acid) afforded the desired product as a beige solid (19.5 mg, 0.03 mmol,
56%).
HRMS-ESI (m/z) [M+H]+ calcd for C29H30FN6O4S2: 609.1749, found 609.1753.
Example 68: 2-({6-[(l,3-Benzothiazol-2-yl)amino]-5-methylpyridazin-3yl}(methyl)amino)-5-[3-(benzyIoxy)azetidin-l-yI]-l,3-thiazole-4-carboxylic acid
Step A: ethyl 5-[3-(benzyloxy)azetidin-l-yl]-2-[methyl(5-methyl-6-{[(2Z)-3-{[2(trimethylsilyl)ethoxy]methyl}-2,3-dihydro-l,3-benzothiazol-2-ylidene]amino}pyridazin-3yl)amino]-l,3-thiazole-4-carboxylate
To an oven dried microwave vial was added the product from Préparation 11b (500 mg, 0.79 mmol, 1 eq), 3-(benzyloxy)azetidine hydrochloride (314 mg, 1.57 mmol, 2 eq), césium carbonate (769 mg, 2.36 mmol, 3 eq), and 1,4-dioxane (10 mL). The mixture was sparged with nitrogen (10 min) before adding Xantphos Pd G3 (74.7 mg, 0.08 mmol, 0.1 eq) and the mixture heated at 140 °C for 2 h under microwave irradiation. The reaction was diluted with ethyl acetate then washed with water followed by brine, and the organic phase was dried (magnésium sulfate) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 24 g RediSep™ silica cartridge) eluting with a gradient of 0 - 50% ethyl acetate in Ao-heptane afforded the desired product as a yellow foam (380 mg, 0.53 mmol, 67%).
LC/MS (C35H43N7O4SiS2) 718 [M+H]+; RT 1.53 (LCMS-V-B1)
Ή NMR (400 MHz, DMSO-d6) 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).
290
Step B: ethyl 2-({6-[(l,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)5-[3-(benzyloxy)azetidin-l-yl]-l,3-thiazole-4-carboxylate
Trifluoroacetic acid (1 mL, 13.4 mmol, 96.3 eq) was added to a stirred solution of the product from Step A (100 mg, 0.14 mmol, 1 eq) in dichloromethane (3 mL) and the mixture was stirred at ambient température overnight. The reaction was diluted with dichloromethane and washed with saturated aqueous sodium bicarbonate then brine, dried (magnésium sulfate), and concentrated in vacuo. Purification by reverse phase automated flash chromatography (CombiFlash Rf, C18 13g RediSep column) eluting with a gradient of 5 - 95% acetonitrile (pH 4; formic acid) in water (pH4; formic acid) afforded the desired product as a yellow glass (48 mg, 0.08 mmol, 59%).
LC/MS (C29H29N7O3S2) 588 [M+H]+; RT 1.31 (LCMS-V-B1)
Ή NMR (400 MHz, DMSO-d6) 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-[(l,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)-5-[3(benzyloxy)azetidin-l-yl]-l,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 overnight. The reaction was allowed to cool to ambient température and concentrated in vacuo. The residue was triturated in water, filtered, and dried under vacuum to afford the desired product as a beige solid (33.2 mg, 0.06 mmol, 73%) [as a lithium sait].
HRMS-ESI (m/z) [M+H]+ calcd for C27H26N7O3S2: 560.1533, found 560.1531.
Example 69: 2-({6-[(l,3-Benzothiazol-2-yl)amino]-5-methylpyridazin-3yl}(methyl)amino)-5-(3-phenoxyazetidin-l-yl)-l,3-thiazole-4-carboxylic acid
291
Step A: ethyl 2-[methyl(5-methyl-6-{[(2Z)-3-{[2-(trimethylsilyl)ethoxy]methyl}-2,3-dihydrol,3-benzothiazol-2-ylidene]amino}pyridazin-3-yl)amino]-5-(3-phenoxyazetidin-l-yl)-l,3thiazole-4-carboxylate
To an oven-dried vial was added the product from Préparation 11b (100 mg, 0.16 mmol, 1 eq), 3-phenoxy-azetidine hydrochloride (58.4 mg, 0.31 mmol, 2 eq), césium carbonate (154 mg, 0.47 mmol, 3 eq), and 1,4-dioxane (4 mL). The mixture was sparged 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 was diluted with ethyl acetate, washed with water followed by brine, dried (magnésium sulfate), and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 12 g RediSep™ silica cartridge) eluting with a gradient of 0 - 50% ethyl acetate in Ao-heptane afforded the desired product as a yellow glass (55.7 mg, 0.08 mmol, 50%).
LC/MS (C34H4iN7O4SiS2) 704 [M+H]+; RT 1.60 (LCMS-V-B1)
Ή NMR (400 MHz, DMSO-d6) 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-[(l,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)5-(3-phenoxyazetidin-l-yl)-l,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 température overnight. The reaction was diluted with dichloromethane, washed with 25 saturated aqueous sodium bicarbonate then brine, dried (magnésium sulfate), and concentrated in vacuo. Purification by reverse phase automated flash chromatography (CombiFlash Rf, C18
292
13g RediSep column) eluting with a gradient of 5 - 95% acetonitrile (pH4; formic acid) in water (pH 4; formic acid) afforded the desired product as a yellow glass (20.4 mg, 0.04 mmol, 45%).
LC/MS (C28H27N7O3S2) 574 [M+H]+; RT 1.31 (LCMS-V-B1)
Al NMR (400 MHz, DMSO-d6) 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-[(l,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)-5-(3phenoxyazetidin-1 -yl)-l,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 lithium hydroxide monohydrate (7.31 mg, 0.17 mmol, 5 eq) and the mixture was heated at 100 °C overnight. The reaction was allowed to cool to ambient température, concentrated in vacuo, triturated with water, filtered, and dried under vacuum to afford the desired product as a beige solid (11.2 mg, 0.02 mmol, 59%) [as a lithium sait].
HRMS-ESI (m/z) [M+H]+ calcd for C26H24N7O3S2: 546.1377, found 546.1381.
Example 70: 2-({6-[(l,3-Benzothiazol-2-yl)amino]-5-methylpyridazin-3yl}(methyl)amino)-5-[(3S)-3-(benzyloxy)pyrrolidin-l-yl]-l,3-thiazole-4-carboxylic acid
Step A: ethyl 5-[(3S)-3-(benzyloxy)pyrrolidin-l-yl]-2-[methyl(5-methyl-6-{[(2Z)-3-{[2(trimethylsilyl)ethoxy]methyl}-2,3-dihydro-l,3-benzothiazol-2-ylidene]amino}pyridazin-3yl)amino]-l,3-thiazole-4-carboxylate
To an oven dried vial was added the product from Préparation 11b (200 mg, 0.31 mmol, 1 eq), (Sj-S-benzyloxy-pyrrolidine hydrochloride (135 mg, 0.63 mmol, 2 eq), césium carbonate (308
293 mg, 0.94 mmol, 3 eq), and 1,4-dioxane (8 mL). The mixture was sparged with nitrogen (10 mm) before the addition of rac-BINAP Pd G3 (15.6 mg, 0.02 mmol, 0.05 eq) and the mixture heated at 100 °C for 24 h. The reaction was diluted with ethyl acetate, then washed with water followed by brine, dried (magnésium sulfate), and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 12 g RediSep™ silica cartridge) eluting with a gradient of 0 - 50% ethyl acetate in Ao-heptane afforded the desired product as a yellow foam (113 mg, 0.15 mmol, 49%).
LC/MS (C36H45N7O4SiS2) 732 [M+H]+; RT 1.54 (LCMS-V-B1) ΧΗ NMR (400 MHz, DMSO-d6) δ 7.82 (d, 1H), 7.57 (d, 1H), 7.49 - 7.40 (m, 2H), 7.36 - 7.31 (m, 4H), 7.31 - 7.22 (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-[(l,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(inethyl)amino)5-[(3 S)-3-(benzyloxy)pyrrolidin-l -yl]-l ,3-thiazole-4-carboxylate
Hydrochloric acid (4M 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 was diluted with ethyl acetate, washed with saturated aqueous sodium bicarbonate then brine, dried (magnésium sulfate), and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 12 g RediSep™ silica cartridge) eluting with a gradient of 0 — 5% methanol in dichloromethane afforded the desired product as a yellow glass (83 mg, 0.14 mmol, 89%).
LC/MS (C30H3iN7O3S2) 602 [M+H]+; RT 1.32 (LCMS-V-B1)
Ή NMR (400 MHz, DMSO-d6) 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-[(l,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)-5[(3S)-3-(benzyloxy)pyrrolidin-l-yl]-l ,3-thiazole-4-carboxylic acid
294
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 overnight. The reaction was allowed to cool to ambient température and concentrated in vacuo, triturated with water, filtered, and dried under vacuum to afford the desired product as a yellow solid (65.6 mg, 0.11 mmol, 83%).
HRMS-ESI (m/z) [M+H]+ calcd for C28H28N7O3S2: 574.1690, found 574.1687.
Example 71: 2-({6-[(l,3-Benzothiazol-2-yl)amino]-5-methylpyridazin-3yl}(methyl)amino)-5-[(3.R)-3-(benzyloxy)pyrrolidin-l-yl]-l,3-thiazole-4-carboxylic acid
Step A: ethyl 5-[(3R)-3-(benzyloxy)pyrrolidin-l-yl]-2-[methyl(5-methyl-6-{[(2Z)-3-{[2(trimethylsilyl)ethoxy]methyl}-2,3-dihydro-l,3-benzothiazol-2-ylidene]amino}pyridazin-3yl)amino]-l,3-thiazole-4-carboxylate
To an oven-dried vial was added the product from Préparation 11b (200 mg, 0.31 mmol, 1 eq), (R)-3-benzyloxy-pyrrolidine hydrochloride (135 mg, 0.63 mmol, 2 eq), césium carbonate (308 mg, 0.94 mmol, 3 eq), and 1,4-dioxane (8 mL). The mixture was sparged with nitrogen (10 min) before the addition of rac-BINAP Pd G3 (31.2 mg, 0.03 mmol, 0.1 eq) and the mixture heated at 100 °C for 24 h. The reaction was diluted with ethyl acetate, washed with water followed by brine, dried (magnésium sulfate), and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 12 g RediSep™ silica cartridge) eluting with a gradient of 0 - 50% ethyl acetate in Ao-heptane afforded the desired product as a yellow glass (101 mg, 0.14 mmol, 44%).
LC/MS (C36H45N7O4SiS2) 732 [M+H]+; RT 1.55 (LCMS-V-B1) * H NMR (400 MHz, DMSO-d6) 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,
295 φ 1Η), 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-[(l,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)5 5-[(3R)-3-(benzyloxy)pyrrolidin-l-yl]-l,3-thiazole-4-carboxylate
Hydrochloric acid (4M 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 was diluted with dichloromethane, washed with saturated aqueous sodium bicarbonate then brine, dried (magnésium sulfate), and concentrated 10 in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 12 g
RediSep™ silica cartridge) eluting with a gradient of 0 - 5% methanol in dichloromethane afforded the desired product as a yellow gum (80 mg, 0.13 mmol, 97%).
LC/MS (C3oH3iN703S2) 602 [M+H]+; RT 1.32 (LCMS-V-B1)
Ή NMR (400 MHz, DMSO-d6) δ 10.98 (br s, 1H), 7.91 (s, 1H), 7.60 (br s + s, 2H), 7.42 15 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-[(l,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl) amino)-5[(3R)-3-(benzyloxy)pyrrolidin-l-yl]-l,3-thiazole-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 at 100 °C overnight. The reaction was allowed to cool to ambient température and concentrated in vacuo, triturated with water, filtered, and dried under vacuum to afford the desired product as a yellow solid (50.1 mg, 0.09 mmol, 65%) [as a lithium sait].
HRMS-ESI (m/z) [M+H]+ calcd for C28H28N7O3S2: 574.1690, found 574.1723.
Example 72: 2-({6-[(l,3-Benzothiazol-2-yl)amino]-5-methylpyridazin-3yl}(methyl)amino)-5-(3-hydroxyazetidin-l-yl)-l,3-thiazole-4-carboxylic acid
296
Step A: ethyl2-({6-[(l,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(inethyl)amino)5-(3-hydroxyazetidin-l-yl)-l,3-thiazole-4-carboxylate
A solution of the product from Example 68, Step B (94.2 mg, 0.16 mmol, 1 eq) in 5 dichloromethane (5 mL) was cooled to -78 °C and boron trichloride (IM in dichloromethane;
0.48 mL, 0.48 mmol, 3 eq) was added slowly. The mixture was maintained at -78 °C for 1 h, then allowed to warm to ambient température over 2 h. The reaction was quenched by the addition of saturated aqueous sodium bicarbonate then extracted with dichloromethane (x2) followed by 3:1 dichloromethane / isopropanol, and the combined organic extracts were dried 10 (magnésium sulfate) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 12 g RediSep™ silica cartridge) eluting with a gradient of 0 - 6% methanol in dichloromethane afforded the desired product as a yellow solid (13.4 mg, 0.03 mmol, 17%).
LC/MS (C22H23N7O3S2) 498 [M+H]+; RT 1.00 (LCMS-V-B1)
Ή NMR (400 MHz, DMSO-d6) 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-[(l,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)-5-(320 hydroxyazetidin-l-yl)-l,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 overnight. The reaction was allowed to cool to ambient température and concentrated in vacuo. Purification by reverse phase automated flash chromatography (CombiFlash Rf, C18 25 13g RediSep column) eluting with a gradient of 5 - 95% acetonitrile in water afforded the desired product as a beige solid (8.1 mg, 0.02 mmol, 64%).
HRMS-ESI (m/z) [M+H]+ calcd for C20H20N7O3S2: 470.1064, found 470.1072.
297
Example 73: 2-({6-[(l,3-Benzothiazol-2-yl)ammo]-5-methylpyridazm-3yl}(methyl)ammo)-5-(3-{2-fluoro-4-[3-(methylamino)prop-l-yn-l-yl]phenoxy}propyl)l,3-thiazole-4-carboxylic acid
\
Step A: ethyl 2-({6-[(l,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)5-(3-{2-fluoro-4-[3-(methylamino)prop-l-yn-l-yl]phenoxy}propyl)-l,3-thiazole-4carboxylate
Trifluoroacetic acid (20 mL) was added to a stirred solution of the product from Préparation 5j, Step A (1.5 g, 1.71 mmol, 1 eq) in dichloromethane (60 mL) and the mixture was stirred at ambient température overnight. The reaction was diluted with dichloromethane, cooled to 0 °C then basified by the addition of 2N aqueous sodium hydroxide, and the organic phase was dried (magnésium sulfate) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 40 g RediSep™ silica cartridge) eluting with a gradient of 0 - 10% methanol in dichloromethane afforded the desired product as a yellow solid (361 mg, 0.56 mmol, 33%).
LC/MS (C32H32FN7O3S2) 646 [M+H]+; RT 1.98 (LCMS-V-C) XH NMR (400 MHz, DMSO-d6) 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).
StepB: 2-({6-[(l,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)-5-(3{2-fluoro-4-[3-(methylamino)prop-l-yn-l-yl]phenoxy}propyl)-l,3-thiazole-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
298 at 100 °C overnight. The reaction was allowed to cool to ambient température and concentrated in vacuo. The residue was triturated with water, filtered, washed with water then diethyl ether, and dried under vacuum to afford the desired product as a yellow solid (286 mg, 0.46 mmol, 83%) [as a lithium sait],
HRMS-ESI (m/z) [M+H]+ calcd for C30H29FN7O3S2: 618.1752, found 618.1767.
Example 74: 2-({6-[(l,3-Benzothiazol-2-yl)ammo]-5-methylpyridazin-3yl}(methyl)amino)-5-{3-[2-fluoro-4-(3-{methyl[2-(methylamino)ethyl]amino}prop-l-ynl-yl)phenoxy]propyl}-l,3-thiazole-4-carboxylic acid
Step A: ethyl5-[3-(4-{3-[(2-{[(tertbutoxy) carbonyl](methyl) amino}ethyl) (methyl) amino]prop-l -yn-1 -yl}-2fluorophenoxy)propyl]-2-[methyl(5-methyl-6-{[(2Z)-3-{[2-(trimethylsilyl)ethoxy]methyl}2,3-dihydro-l,3-benzothiazol-2-ylidene]amino}pyridazin-3-yl)amino]-l,3-thiazole-4carboxylate
To a solution of the product from Préparation 5j (523 mg, 0.67 mmol, 1 eq) in acetonitrile (30 mL) was added A-Boc-(methylamino)acetaldehyde (234 mg, 1.35 mmol, 2 eq), sodium triacetoxyborohydride (343 mg, 1.62 mmol, 2.4 eq) and glacial acetic acid (50 pL) and the mixture was stirred at ambient température for 6 h. The reaction was partitioned between ethyl acetate and saturated aqueous sodium bicarbonate and the organic phase was dried (magnésium sulfate) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 40 g RediSep™ silica cartridge) eluting with a gradient of 0 - 50% ethyl acetate in zso-heptane afforded the desired product as a yellow gum (705 mg, 0.76 mmol, > 100%).
299
LC/MS (C46H6iN8O6FSiS2) 933 [M+H]+; RT 1.39 (LCMS-V-B2)
Ή NMR (400 MHz, DMSO-d6) 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-[(l,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)5-{3-[2-fluoro-4-(3-{methyl[2-(methylamino)ethyl]amino}prop-l-yn-l-yl)phenoxy]propyl}l,3-thiazole-4-carboxylate
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 température for 12 h. The reaction was diluted with dichloromethane, cooled to 0 °C and basified by the addition of 2N aqueous sodium hydroxide, and the organic phase was dried (magnésium sulfate) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 12 g RediSep™ silica cartridge) eluting with a gradient of dichloromethane to 20% methanol in dichloromethane afforded the desired product as a yellow gum (123 mg, 0.17 mmol, 22%).
LC/MS (C35H39FN8O3S2) 703 [M+H]+; RT 2.07 (LCMS-V-C)
Hl NMR (400 MHz, DMSO-d6) 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), 2.19 - 2.08 (m, 2H), 1.30 (t, J = 7.1 Hz, 3H).
Step C: 2-({6-[(l,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)-5-{3[2-fluoro-4-(3-{methyl[2-(methylamino)ethyl]amino}prop-l-yn-l-yl)phenoxy]propyl}-l,3thiazole-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 at 100 °C overnight. The reaction was allowed to cool to ambient température and concentrated in vacuo. Purification by reverse phase automated flash chromatography (CombiFlash Rf, C18 13g RediSep column) eluting with a gradient of 5 - 95% acetonitrile (pH 9; ammonium formate) in water (pH 9; ammonium formate) afforded the desired product as a yellow solid (75.8 mg, 0.11 mmol, 64%) [as a lithium sait].
300
λ
HRMS-ESI (m/z) [M+H]+ calcd for C33H36FN8O3S2: 675.2330, found 675.2331.
Example 75: 2-({6-[(l,3-Benzothiazol-2-yl)amino]-5-methylpyridazin-3yl}(methyl)amino)-5-[4-(benzyloxy)piperidin-l-yl]-l,3-thiazole-4-carboxylic acid
Step A: ethyl 5-[4-(benzyloxy)piperidin-l-yl]-2-[(6-chloro-5-methylpyridazùi-3yl)(methyl)amino]-l,3-thiazole-4-carboxylate
Sodium Hydride (60% in minerai oil; 27.2 mg, 0.68 mmol, 1.2 eq) was added to a cooled solution of the product from Préparation 3za (213 mg, 0.57 mmol, 1 eq) and 3,6-dichloro-4methylpyridazine (92.5 mg, 0.57 mmol, 1 eq) in tetrahydrofuran (6 mL) and the mixture was 10 allowed to warm to ambient température and stir overnight. The reaction was diluted with dichloromethane, washed with saturated aqueous ammonium chloride then brine, dried (magnésium sulfate), and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 24 g RediSep™ silica cartridge) eluting with a gradient of 0 - 60% ethyl acetate / Ao-heptane afforded the desired product as a yellow solid (101 mg, 0.2 15 mmol, 35%).
LC/MS (C24H28CIN5O3S) 502 [M+H]+; RT 1.29 (LCMS-V-B1) XH NMR (400 MHz, DMSO-d6) 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 20 —1.63 (m, 2H), 1.30 (t, J = 7.1 Hz, 3H).
Step B: ethyl 2-({6-[(l,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)5-[4-(benzyloxy)piperidin-l-yl]-l,3-thiazole-4-carboxylate
301
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), A,7V-dnsopropylethylamine (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 sparged with nitrogen (10 min) then tris(dibenzylideneacetone)dipalladium(0) (9.19 mg, 0.01 mmol, 0.05 eq) was added and the mixture was heated in a sealed flask at 150 °C overnight. The reaction was allowed to cool to ambient température and concentrated in vacuo. Purification by reverse phase automated flash chromatography (CombiFlash Rf, C18 13g RediSep column) eluting with a gradient of 5 - 95% acetonitrile (pH4; formic acid) in water (pH 4; formic acid) afforded the desired product as a yellow glass (31.2 mg, 0.05 mmol, 25%).
LC/MS (C31H33N7O3S2) 616 [M+H]+; RT 1.36 (LCMS-V-B1)
Hl NMR (400 MHz, DMSO-d6) 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), 3.08 - 2.97 (m, 2H), 2.46 (s, 3H), 2.08 - 1.97 (m, 2H), 1.80 - 1.69 (m,2H), 1.32 (t, 3H).
Step Q: 2-({6-[(l,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)-5-[4(benzyloxy)piperidin-l -yl]-l ,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 overnight. The reaction was allowed to cool to ambient température and concentrated in vacuo. Purification by reverse phase automated flash chromatography (CombiFlash Rf, C18 13g RediSep column) eluting with a gradient of 5 - 95% acetonitrile in water afforded the desired product as a cream solid (12.2 mg, 0.02 mmol, 41%) [as a lithium sait].
HRMS-ESI (m/z) [M+H]+ calcd for C29H30N7O3S2: 588.1846, found 588.1854.
Example 76: 2-({6-[(l,3-Benzothiazol-2-yl)amino]-5-methylpyridazin-3yl}(methyl)amino)-5-[3-(2-fluoro-4-{3-[(2-hydroxyethyl)(methyl)amino]prop-l-yn-lyl}phenoxy)propyl]-l,3-thiazole-4-carboxy!ic acid
302
Step A: ethyl 5-(3-{4-[3-({2-[(tert-butyldimethylsilyl)oxy]ethyl}(methyl)amino)prop-l-yn-1 yl]-2-fluorophenoxy}propyl)-2-[methyl(5-methyl-6-{[(2Z)-3-{[2(trimethylsilyl)ethoxy]methyl}-2,3-dihydro-l,3-benzothiazol-2-ylidene]amino}pyridazin-35 yl)amino]-l,3-thiazole-4-carboxylate
To a solution of the product from Préparation 5j (747 mg, 0.96 mmol, 1 eq) in acetonitrile (50 mL) was added (iert-butyldimethylsiloxy)acetaldehyde (367 pL, 1.93 mmol, 2 eq), followed by sodium triacetoxyborohydride (612 mg, 2.89 mmol, 3 eq) and glacial acetic acid (50 pL) and the mixture was stirred at ambient température overnight. The reaction was partitioned 10 between ethyl acetate and saturated aqueous sodium bicarbonate, and the organic phase was dried (magnésium sulfate), and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 24 g RediSep™ silica cartridge) eluting with a gradient of 0 - 60% ethyl acetate in zso-heptane afforded the desired product as a clear oil (907 mg, 0.96 mmol, 100%).
LC/MS (C46H64FN7O5Si2S2) 934 [M+H]+; RT 1.47 (LCMS-V-B2) XH NMR (400 MHz, DMSO-d6) δ 7.84 (d, 1H), 7.67 (d, 1H), 7.50 - 7.40 (m 2H), 7.31 - 7.22 (m, 2H), 7.21 - 7.12 (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-[(l,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)5-[3-(2-fluoro-4-{3-[(2-hydroxyethyl)(methyl)amino]prop-l-yn-l-yl}phenoxy)propyl]-l,3thiazole-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 25 température for 9 h. The reaction was diluted with dichloromethane, cooled to 0 °C and basified by the addition of 2N aqueous sodium hydroxide, and the organic phase was dried (magnésium
303
V sulfate) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 12 g RediSep™ silica cartridge) eluting with a gradient of 0 - 20% methanol in dichloromethane afforded the desired product as a yellow solid (95 mg, 0.14 mmol, 14%).
LC/MS (C34H36FN7O4S2) 690 [M+H]+; RT 1.98 (LCMS-V-C)
Ή NMR (400 MHz, DMSO-d6) 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-[(l,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)-5-[310 (2-fluoro-4-{3-[(2-hydroxyethyl)(methyl)amino]prop-l-yn-l-yl}phenoxy)propyl]-l,3thiazole-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 h. The reaction was allowed to cool to ambient température and concentrated in 15 vacuo. Purification by reverse phase automated flash chromatography (CombiFlash Rf, C18 13g RediSep column) eluting with a gradient of 5 - 95% acetonitrile (pH 9; ammonium acetate) in water (pH 9; ammonium acetate) afforded the desired product as a yellow solid (61.8 mg, 0.09 mmol, 67%).
HRMS-ESI (m/z) [M+H]+ calcd for C32H33FN7O4S2: 662.2014, found 662.2035.
Example 77: 2-({6-[(l,3-Benzothiazol-2-yl)amino]-5-methylpyridazin-3yl}(methyl)amino)-5-[(3S')-3-phenoxypyrrolidin-l-yl]-l,3-thiazole-4-carboxylic acid
Step A: ethyl2-[methyl(5-methyl-6-{[(2Z)-3-{[2-(trimethylsilyl)ethoxy]methyl}-2,3-dihydrol,3-benzothiazol-2-ylidene]amino}pyridazin-3-yl)amino]-5-[(3S)-3-phenoxypyrrolidin-l-yl]25 l,3-thiazole-4-carboxylate
304
To an oven-dried vial was added the product from Préparation 11b (200 mg, 0.31 mmol, 1 eq), (SSj-S-phenoxypyrrolidine hydrochloride (0.09 mL, 0.63 mmol, 2 eq), césium carbonate (308 mg, 0.94 mmol, 3 eq) and 1,4-dioxane (8 mL). The mixture was sparged with nitrogen (10 min) then rac-BINAP Pd G3 (31.2 mg, 0.03 mmol, 0.1 eq) was added and the mixture heated at 100 °C for 6 h. The reaction was diluted with ethyl acetate then washed with water followed by brine, dried (magnésium sulfate), and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 12 g RediSep™ silica cartridge) eluting with a gradient of 0 - 50% ethyl acetate in zso-heptane afforded the desired product as a yellow foam (184 mg, 0.26 mmol, 81%).
LC/MS (C35H43N7O4SiS2) 718 [M+H]+; RT 1.54 (LCMS-V-B1) *H NMR (400 MHz, DMSO-d6) δ 7.82 (d, 1H), 7.58 (d, 1H), 7.48 - 7.37 (m, 2H), 7.35 - 7.20 (m, 4H), 7.01 - 6.91 (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), 3.77 - 3.60 (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-[(l,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)5-[(3S)-3-phenoxypyrrolidin-l-yl]-l,3-thiazole-4-carboxylate
Hydrochloric acid (4M 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 was diluted with dichloromethane, washed with saturated aqueous sodium bicarbonate then brine, dried (magnésium sulfate) and concentrated in vacuo. Purification by reverse phase automated flash chromatography (CombiFlash Rf, C18 13g RediSep column) eluting with 5 - 95% acetonitrile (pH4; formic acid) in water (pH 4; formic acid) afforded the desired product as a yellow gum (55.8 mg, 0.09 mmol, 37%).
LC/MS (C29H29N7O3S2) 588 [M+H]+; RT 1.32 (LCMS-V-B1) JH NMR (400 MHz, DMSO-d6) 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-[(l,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)-5[(3S)-3-phenoxypyrrolidin-l-yl]-l,3-thiazole-4-carboxylic acid
305
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 overnight. The reaction was allowed to cool to ambient température, concentrated in vacuo, and the residue was triturated with water, filtered, and dried under vacuum to afford the desired product as a yellow solid (38.5 mg, 0.07 mmol, 72%) [as a lithium sait],
HRMS-ESI (m/z) [M+H]+ calcd for C27H26N7O3S2: 560.1533, found 560.1541.
Example 78: 2-({6-[(l,3-Benzothiazol-2-yl)amino]-5-methylpyridazin-3yl}(methyl)amino)-5-cyclobutyl-l,3-thiazole-4-carboxylic acid
Step A: ethyl 5-cyclobutyl-2-[methyl(5-rnethyl-6-{[(2Z)-3-{[2-(trimethylsilyl)ethoxy]methyl}2,3-dihydro-l,3-benzothiazol-2-ylidene]amino}pyridazin-3-yl)amino]-l,3-thiazole-4carboxylate
To an oven-dried flask was added the product from Préparation 11b (500 mg, 0.79 mmol, 1 eq) and copper(I) iodide (300 mg, 1.57 mmol, 2 eq) and the flask was evacuated and flushed with nitrogen (x3) and then dimethylacetamide (15 mL) was added. Cyclobutylzinc bromide (0.5M in tetrahydrofuran; 9.44 mL, 4.72 mmol, 6 eq) was added in one portion and reaction stirred overnight at ambient température. The mixture was diluted with dichloromethane, then quenched with saturated aqueous sodium bicarbonate causing précipitation. The mixture was stirred vigorously stirred for 5 min, then filtered through a dichloromethane-wet celite pad, and eluted through 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 (magnésium sulfate) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 24 g RediSep™ silica cartridge) eluting with a gradient of 0 — 30% ethyl acetate in Ao-heptane afforded the desired product as a pale yellow glass/ foam (313 mg, 0.51 mmol, 65%).
LC/MS (C29H38N6O3SiS2) 611 [M+H]+; RT 1.38 (LCMS-V-B2)
306 “ XH NMR (400 MHz, CDC13) 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), 2.38 - 2.16 (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-[(l,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)-5cyclobutyl-l,3-thiazole-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 température overnight. The reaction was concentrated in vacuo. 1,4-dioxane (10 10 mL) was added with stirring to afford a thick off-white suspension, to which was added IN aqueous sodium hydroxide (5 mL) and the mixture was heated at reflux for 20 min. The mixture was allowed to cool to ambient température, then filtered through a pad of celite, eluting through with water then dioxane and concentrated in vacuo. The residue was partitioned between chloroform and saturated aqueous sodium bicarbonate, and the organic phase was 15 dried (magnésium sulfate) and concentrated in vacuo. The solid material was triturated in diethyl ether, filtered, and dried under vacuum to afford the desired product as a yellow powder (98.5 mg, 0.22 mmol, 43%).
HRMS-ESI (m/z) [M+H]+ calcd for C21H21N6O2S2: 453.1162, found 453.1172.
Example 79: 2-({6-[(l,3-Benzothiazol-2-yl)amino]-5-methylpyridazin-320 yI}(methyl)amino)-5-[(3B)-3-phenoxypyrrolidin-l-yl]-l,3-thiazoIe-4-carboxylic acid
Step A: ethyl2-[methyl(5-methyl-6-{[(2Z)-3-{[2-(trimethylsilyl)ethoxy]methyl}-2,3-dihydrol,3-benzothiazol-2-ylidene]amino}pyridazin-3-yl)amino]-5-[(3R)-3-phenoxypyrrolidin-lyl]-l,3-thiazole-4-carboxylate
307
To an oven-dried flask was added the product from Préparation 11b (300 mg, 0.44 mmol, 1 eq), (R)-3-phenoxy-pyrrolidine (0.13 mL, 0.88 mmol, 2 eq), césium carbonate (430 mg, 1.32 mmol, 3 eq) and 1,4-dioxane (12 mL). The mixture was sparged with nitrogen (10 min) and racBINAP Pd G3 (43.6 mg, 0.04 mmol, 0.1 eq) was added and the mixture heated at 100 °C 5 overnight. The reaction was diluted with ethyl acetate then washed with water followed by brine, dried (magnésium sulfate), and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 12 g RediSep™ silica cartridge) eluting with a gradient of 0 - 50% ethyl acetate in Ao-heptane afforded the desired product as a yellow foam (253 mg, 0.35 mmol, 80%).
LC/MS (C35H43N7O4SiS2) 718 [M+H]+; RT 1.54 (LCMS-V-B1) JH NMR (400 MHz, DMSO-d6) 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 15 - 2.16 (m, 1H), 1.27 (t, J = 7.1 Hz, 3H), 0.96 - 0.83 (m, 2H), -0.11 (s, 9H).
Step B: ethyl 2-({6-[(l,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)5-[(3R)-3-phenoxypyrrolidin-l-yl]-l,3-thiazole-4-carboxylate
Hydrochloric acid (4M 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 20 was stirred at 50 °C overnight. The reaction was diluted with dichloromethane, washed with saturated aqueous sodium bicarbonate then brine, dried (magnésium sulfate), and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 12 g RediSep™ silica cartridge) eluting with a gradient of 0 - 5% methanol in dichloromethane afforded the desired product as a yellow foam (189 mg, 0.32 mmol, 91%).
LC/MS (C29H29N7O3S2) 588 [M+H]+; RT 1.32 (LCMS-V-B1) XH NMR (400 MHz, DMSO-d6) 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), 30 2.26 - 2.17 (m, 1H), 1.27 (t, J = 7.1 Hz, 3H).
Step C: 2-({6-[(l,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)-5[(3R)-3-phenoxypyrrolidin-l-yl]-l,3-thiazole-4-carboxylic acid
308
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 heated at reflux overnight. The reaction was allowed to cool to ambient température then concentrated in vacuo, triturated with water, filtered, and dried under vacuum to afford the desired product 5 as a beige solid (118 mg, 0.21 mmol, 65%) [as a lithium sait].
HRMS-ESI (m/z) [M+H]+ calcd for C27H26N7O3S2: 560.1533, found 560.1535.
Example 80: 2-({6-[(l,3-Benzothiazol-2-yl)amino]-5-methylpyridazin-3yl}(methyl)amino)-5-(piperidin-l-yl)-l,3-thiazoIe-4-carboxyIic acid
Step A: ethyl 5-(piperidin-l-yl)-l,3-thiazole-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 l,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 was allowed to cool to ambient température then concentrated in vacuo and purification by automated flash column 15 chromatography (CombiFlash Rf, 24 g RediSep™ silica cartridge) eluting with a gradient of 0 - 60% ethyl acetate in zso-heptane afforded the desired product as a brown oil (376 mg, 1.57 mmol, 74%).
LC/MS (C11H16N2O2S) 241 [M+H]+; RT 1.12 (LCMS-V-B1)
Ή NMR (400 MHz, DMSO-d6) δ 8.45 (s, 1H), 4.24 (q, J = 7.1 Hz, 2H), 3.15 - 3.07 (m, 4H), 20 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-l-yl)-l ,3-thiazole-4-carboxylate
A-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 température for 7 h. The reaction was partitioned between 10% aqueous sodium 25 thiosulfate and ethyl acetate, and the organic phase was washed with water followed by brine,
309 dried (magnésium sulfate), and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 24 g RediSep™ silica cartridge) eluting with a gradient of 0 - 40% ethyl acetate in Ao-heptane afforded the desired product as a colourless oil (482 mg, 1.51 mmol, 71%).
LC/MS (CiiHi5BrN2O2S) 273 [M - OEt ]+; RT 1.14 (LCMS-V-B1) XH NMR (400 MHz, DMSO-d6) 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-l-yl)-l,3-thiazole-4carboxylate
The product from Step B (482 mg, 1.51 mmol, 1 eq) and ZerAbutyl methylcarbamate (0.26 mL, 1.81 mmol, 1.2 eq) were added 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) under a nitrogen atmosphère. Césium carbonate (738 mg, 2.26 mmol, 1.5 eq) was added and the mixture heated at 100 °C for 7.5 h. The solution was allowed to cool to ambient température and concentrated in vacuo. The mixture was diluted with ethyl acetate and washed with water followed by brine, dried (magnésium sulfate), and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 50 g RediSep™ silica cartridge) eluting with a gradient of 0 - 50% ethyl acetate in Ao-heptane afforded the desired product as a yellow gum (434 mg, 1.17 mmol, 78%).
LC/MS (C17H27N3O4S) 370 [M+H]+; RT 1.304 (LCMS-V-B1)
Ή NMR (400 MHz, DMSO-d6) 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-l-yl)-l,3-thiazole-4-carboxylate
Trifluoroacetic acid (1.32 mL, 17.6 mmol, 15 eq) was added to a stirred solution of the product from Step C (434 mg, 1.17 mmol, 1 eq) in dichloromethane (15 mL) at 0 °C and the mixture was allowed to warm to ambient température and stir overnight. The reaction was partitioned between saturated aqueous sodium bicarbonate and dichloromethane, and the organic phase was washed with brine, dried (magnésium sulfate), and concentrated in vacuo. The résultant solid was triturated with diethyl ether, collected by filtration and dried under vacuum to afford the desired product as a white solid (197 mg, 0.73 mmol, 62%).
LC/MS (Ci2Hi9N3O2S) 270 [M+H]+; RT 0.91 (LCMS-V-B1)
310 XH NMR (400 MHz, DMSO-d6) 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: ethyl2-[(6-chloro-5-methylpyridazin-3-yl)(methyl)amino]-5-(piperidin-l-yl)-l,3thiazole-4-carboxylate
Sodium Hydride (60% in minerai 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-4methylpyridazine (119 mg, 0.73 mmol, 1 eq) in tetrahydrofuran (8 mL) and the mixture was allowed to warm to ambient température and stirred for 3.5 h. The reaction was diluted with dichloromethane, washed with saturated aqueous ammonium chloride followed by brine, dried (magnésium sulfate), and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 24 g RediSep™ silica cartridge) eluting with a gradient of 0 - 60% ethyl acetate in zso-heptane afforded the desired product as a yellow solid (131 mg, 0.33 mmol, 45%).
LC/MS (Ci7H22C1N5O2S) 396 [M+H]+; RT 1.20 (LCMS-V-B1)
Al NMR (400 MHz, DMSO-d6) 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-[(l,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)5-(piperidin-l -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), lV,7V-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 sparged 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 sealed flask at 150 °C overnight. The reaction was allowed to cool to ambient température then partitioned between ethyl acetate and water, and the organic phase was washed with brine, dried (magnésium sulfate), and concentrated in vacuo. Purification by reverse phase automated flash chromatography (CombiFlash Rf, C18 13g RediSep column) eluting with a gradient of 5 - 95% acetonitrile (pH4; formic acid) in water (pH 4; formic acid) afforded the desired product as a yellow foam (64.8 mg, 0.13 mmol, 38%).
LC/MS (C24H27N7O2S2) 510 [M+H]+; RT 1.32 (LCMS-V-B1)
311
Ή NMR (400 MHz, DMSO-d6) 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-[(l,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)-5(piperidin-1 -yl)-l ,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 heated at reflux overnight. The reaction was allowed to cool to ambient température and concentrated in vacuo. Purification by reverse phase automated flash chromatography (CombiFlash Rf, C18 13g RediSep column) eluting with a gradient of 5 - 95% acetonitrile in water afforded the desired product as a yellow solid (23 mg, 0.05 mmol, 38%).
HRMS-ESI (m/z) [M+H]+ calcd for C22H24N7O2S2: 482.1427, found 482.1435.
Example 81: 2-({6-[(l,3-Benzothiazol-2-yl)amino]-5-methylpyridazin-3yl}(methyl)amino)-5-(4-phenoxypiperidin-l-yl)-l,3-thiazole-4-carboxyIic acid
Step A: ethyl 5-(4-phenoxypiperidin-l-yl)-l,3-thiazole-4-carboxylate
A solution of ethyl 5-bromothiazole-4-carboxylate (500 mg, 2.12 mmol, 1 eq), 4phenoxypiperidine (451 mg, 2.54 mmol, 1.2 eq) and l,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 température and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 40 g RediSep™ silica cartridge) eluting with a gradient of 0 - 60% ethyl acetate in Ao-heptane afforded the desired product as a brown oil (424 mg, 1.27 mmol, 60%).
LC/MS (C17H20N2O3S) 333 [M+H]+; RT 1.28 (LCMS-V-B1)
312
Ή NMR (400 MHz, DMSO-d6) 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-l-yl)-l,3-thiazole-4-carboxylate •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 température for 5.5 h. The reaction was quenched by the addition of 10% aqueous sodium thiosulfate, then extracted with ethyl acetate, washed with water followed by brine, dried (magnésium sulfate), and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 12 g RediSep™ silica cartridge) eluting with a gradient of 0 - 40% ethyl acetate in Ao-heptane afforded the desired product as a white solid (404 mg, 0.98 mmol, 77%).
LC/MS (Ci7Hi9BrN2O3S) 413 [M+H]+; RT 1.26 (LCMS-V-B1) XH NMR (400 MHz, DMSO-d6) 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-l-yl)-l,3thiazole-4-carboxylate
The product from Step B (404 mg, 0.98 mmol, 1 eq) and Ze/7-butyl methylcarbamate (0.17 mL, 1.18 mmol, 1.2 eq) were added 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) under a nitrogen atmosphère. Césium carbonate (480 mg, 1.47 mmol, 1.5 eq) was added and the mixture was heated at 100 °C overnight. The reaction was allowed to cool to ambient température then concentrated in vacuo. The residue was partitioned between ethyl acetate and water, and the organic phase was washed with brine, dried (magnésium sulfate), and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 24 g RediSep™ silica cartridge) eluting with a gradient of 0 - 50% ethyl acetate in Aoheptane afforded the desired product as a yellow gum (285 mg, 0.62 mmol, 63%).
LC/MS (C23H3iN3O5S) 462 [M+H]+; RT 1.37 (LCMS-V-B1) XH NMR (400 MHz, DMSO-d6) 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
313 (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-l-yl)-l,3-thiazole-4-carboxylate
Trifluoroaœtic acid (0.46 mL, 6.17 mmol, 10 eq) was added to a stirred solution of the product from Step C (285 mg, 0.62 mmol, 1 eq) in dichloromethane (8 mL) at 0 °C and the mixture was allowed to warm to ambient température and stir overnight. The reaction was quenched by the addition of saturated aqueous sodium bicarbonate, extracted with dichloromethane, and the organic extract washed with brine, dried (magnésium sulfate), and concentrated in vacuo. The résultant solid was triturated with diethyl ether / heptane, filtered, and dried under vacuum to afford the desired product as a cream solid (161 mg, 0.45 mmol, 72%).
LC/MS (C18H23N3O3S) 362 [M+H]+; RT 1.08 (LCMS-V-B1)
Ή NMR (400 MHz, DMSO-d6) 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 (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-lyl)-l ,3-thiazole-4-carboxylate
Sodium Hydride (60% in minerai 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-4methylpyridazine (72.6 mg, 0.45 mmol, 1 eq) in tetrahydrofuran (5 mL) and the mixture was allowed to warm to ambient température and stir overnight. The reaction was diluted with dichloromethane, then washed with saturated aqueous ammonium chloride followed by brine, dried (magnésium sulfate), and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 12 g RediSep™ silica cartridge) eluting with a gradient of 0 - 60% ethyl acetate in Ao-heptane afforded the desired product as a yellow foam (63.3 mg, 0.13 mmol, 29%).
LC/MS (C23H26CIN5O3S) 488 [M+H]+; RT 1.289 (LCMS-V-B1) XH NMR (400 MHz, DMSO-d6) 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).
314
Step F: ethyl 2-({6-[(l,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(niethyl)amino)5-(4-phenoxypiperidin-l-yl)-l,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), A,A-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 sparged with nitrogen (10 min) then trîs(dibenzylideneacetone)dipalladium(0) (5.94 mg, 0.01 mmol, 0.05 eq) was added and the mixture was heated in a sealed flask at 150 °C overnight. The reaction was allowed to cool to ambient température then partitioned between ethyl acetate and water, and the organic phase washed with brine, dried (magnésium sulfate), and concentrated in vacuo. Purification by reverse phase automated flash chromatography (CombiFlash Rf, C18 13g RediSep column) eluting with a gradient of 5 - 95% acetonitrile (pH 4; formic acid) in water (pH 4; formic acid) afforded the desired product as a yellow foam (23 mg, 0.04 mmol, 30%).
LC/MS (C30H31N7O3S2) 602 [M+H]+; RT 1.37 (LCMS-V-B1) !H NMR (400 MHz, DMSO-d6) 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-[(l,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)-5-(4phenoxypiperidin-1 -yl)-l,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 heated at reflux overnight, then allowed to cool to ambient température and concentrated in vacuo. Purification by reverse phase automated flash chromatography (CombiFlash Rf, C18 13g RediSep column) eluting with a gradient of 5 - 95% acetonitrile in water afforded the desired product as a yellow solid (3.7 mg, 0.01 mmol, 17%).
HRMS-ESI (m/z) [M+H]+ calcd for C28H28N7O3S2: 574.1690, found 574.1697.
Example 82: 2-({6-[(l,3-Benzothiazol-2-yl)amino]-5-methylpyridazin-3yl}(methyl)amino)-5-{3-[4-(3-{[(terributoxy)carbonyl](methyl)amino}prop-l-yn-l-yl)-2fluorophenoxy]propyl}-l,3-thiazole-4-carboxyiic acid
315
Step A: ethyl 2-({6-[(l,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)5-(3-iodopropyl)-1,3-thiazole-4-carboxylate
To a solution of the product from Préparation 5g (176 mg, 0.24 mmol, 1 eq) in dichloromethane 5 (5 mL) was added trifluoroacetic acid (5 mL) and the mixture was stirred at ambient température for 4 h. The reaction was diluted with dichloromethane, washed with 2N aqueous sodium hydroxide, dried (magnésium sulfate) and concentrated in vacuo. The residue was triturated with acetonitrile, filtered, and dried under vacuum to afford the desired product as a white solid (93 mg, 0.16 mmol, 64%).
LC/MS (C22H23IN6O2S2) 595 [M+H]+; RT 2.63 (LCMS-V-C) ]H NMR (400 MHz, DMSO-d6) 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), 2.20 - 2.11 (m, 2H), 1.34 (t, J = 7.1 Hz, 3H).
Step B: ethyl 2-({6-[(l,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)15 5-{3-[4-(3-{[(tert-butoxy)carbonyl](methyl)amino}prop-l-yn-l-yl)-2-fluorophenoxy]propyl}1,3-thiazole-4-carboxylate
To a solution of the product from Step A (93 mg, 0.16 mmol, 1 eq) in dimethylformamide (5 mL) was added the product from Préparation 6a (56.8 mg, 0.2 mmol, 1.3 eq) and césium carbonate (153 mg, 0.47 mmol, 3 eq) and the mixture was heated at 80 °C overnight, then 20 allowed to cool to ambient température and concentrated in vacuo. The residue was partitioned between ethyl acetate and brine, and the organic phase was dried (magnésium sulfate) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 12 g RediSep™ silica cartridge) eluting with a gradient of 0 - 100% ethyl acetate in isoheptane afforded the desired product as a yellow gum (45 mg, 0.06 mmol, 39%).
LC/MS (C37H40FN7O5S2) 747 [M+H]+; RT 2.77 (LCMS-V-C)
316
Ή NMR (400 MHz, DMSO-d6) 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-[(l,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)-5-{3[4-(3-{[(tert-butoxy) carbonyl] (methyl) amino}prop-l -yn -1 -yl) -2-fluoroph enoxy]propyl}-l ,3thiazole-4-carboxylic 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 heated at reflux overnight. The reaction was allowed to cool to ambient température and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 4 g RediSep™ silica cartridge) eluting with a gradient of 0 - 10% methanol in dichloromethane afforded the desired product as a yellow solid (5.1 mg, 0.01 mmol, 12%).
HRMS-ESI (m/z) [M+H]+ calcd for C35H37FN7O5S2: 718.2276, found 718.2284.
Example 83: 2-({6-[(l,3-Benzothiazol-2-yl)amino]-5-methylpyridazin-3yI}(methyl)amino)-5-(piperidin-4-yl)-l,3-thiazole-4-carboxylic acid
Step A: ethyl 2-({6-[(l,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)5-(piperidin-4-yl)-l,3-thiazole-4-carboxylate
Trifluoroacetic acid (1 mL, 13.4 mmol, 166 eq) was added to a stirred solution of the product from Préparation 5k, Step B (60 mg, 0.08 mmol, 1 eq) in dichloromethane (4 mL) and the mixture was stirred at ambient température overnight. The reaction mixture was loaded onto an SCX-2 cartridge (5 g, preconditioned with dichloromethane, then methanol), then washed with dichloromethane followed by methanol. Elution with 3.5N methanolic ammonia and concentration in vacuo afforded the desired product as a yellow solid (40.9 mg, 0.08 mmol, 99%).
317
LC/MS (C24H27N7O2S2) 510 [M+H]+; RT 0.857 (LCMS-V-B1)
Ή NMR (400 MHz, DMSO-d6) 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 5 Hz, 3H), 2.00 - 1.92 (m, 2H), 1.63 (qd, J = 12.4, 3.9 Hz, 2H), 1.32 (t, J = 7.1 Hz, 3H).
Step B: 2-({6-[(l,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 heated at reflux overnight. The reaction was allowed to cool to ambient température then concentrated in vacuo. Purification by reverse phase automated flash chromatography (CombiFlash Rf, C18 13g RediSep column) eluting with a gradient of 5 - 95% acetonitrile (pH 4; formic acid) in water (pH 4; formic acid) afforded a solid that was triturated with chloroform, filtered, and dried under vacuum to afford the desired product as a yellow solid (27.4 mg, 0.06 mmol, 71%).
LC/MS (C22H23N7O2S2) 482 [M+H]+; RT 0.772 (LCMS-V-B1) rH NMR (400 MHz, DMSO-d6) 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-ESI (m/z) [M+H]+ calcd for C22H23N7O2S2: 482.1427, found 482.1430.
Example 84: 2-({6-[(l,3-Benzothiazol-2-yl)amino]-5-methylpyridazin-3yl}(methyI)amino)-5-{l-[(terLbutoxy)carbonyl]piperidin-4-yl}-l,3-thiazole-4-carboxylic acid
318
Step A: tert-butyl 4-[2-({6-[(l,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3yl}(methyl)amino)-4-(ethoxycarbonyl)-l,3-thiazol-5-yl]piperidine-l-carboxylate
Tetrabutylammonium fluoride (IM in tetrahydrofuran, 0.48 mL, 0.48 mmol, 6 eq) was added to a stirred solution of the product from Préparation 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 température, partitioned between ethyl acetate and saturated aqueous sodium bicarbonate, and the organic phase was dried (magnésium sulfate) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 12 g RediSep™ silica cartridge) eluting with a gradient of 0 - 10% methanol in dichloromethane afforded the desired product as a yellow glass (37.7 mg, 0.06 mmol, 76%).
LC/MS (C29H35N7O4S2) 610 [M+H]+; RT 1.356 (LCMS-V-B1) XH NMR (400 MHz, DMSO-d6) 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), 2.05 - 1.94 (m, 2H), 1.59 - 1.46 (m, 2H), 1.44 (s, 9H), 1.35 (t, 3H).
Step B: 2-({6-[(l,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)-5-{l[(tert-butoxy) carbonyl]piperidin-4-yl}-l,3-thiazole-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 heated at reflux overnight. The reaction was allowed to cool to ambient température and concentrated in vacuo. Purification by reverse phase automated flash chromatography (CombiFlash Rf, C18 13g RediSep column) eluting with a gradient of 5 - 95% acetonitrile (pH 4; formic acid) in water (pH 4; formic acid) afforded the desired product as a yellow solid (23 mg, 0.04 mmol, 51%).
HRMS-ESI (m/z) [M+H]+ calcd for C27H31N7O4S2: 582.1952, found 582.1956.
Example 85: 2-[[6-(l,3-BenzothiazoI-2-ylamino)-5-methyl-pyridazin-3-yl]-(3,4dihydroxybutyl)amino]thiazole-4-carboxylic acid
319
Step A: methyl 2-[tert-butoxycarbonyl-[2-(2,2-dimethyl-l,3-dioxolan-4yl) ethyl]amino]thiazole-4-carboxylate
Using Mitsunobu General Procedure starting from methyl 2-(tertbutoxycarbonylamino)thiazole-4-carboxylate and 2-(2,2-dimethyl-l,3-dioxolan-4-yl)ethanol as starting materials, 3.07 g (99%) of the desired product was obtained.
Ή NMR (500 MHz, DMSO-d6) 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); HRMS-ESI (m/z): [M+H]+ calcd for Ci7H26N2O6S: 387.1589, found: 387.1585.
Step B: methyl 2-[2-(2,2-dimethyl-l,3-dioxolan-4-yl)ethylamino]thiazole-4-carboxylate
Using Deprotection with HFIP General Procedure starting from the product from Step A as the appropriate carbamate, 1.21 g (60%) of the desired product was obtained.
Ή NMR (500 MHz, DMSO-de) 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); HRMS-ESI (m/z): [M+H]+ calcd for Ci2Hi8N2O4S: 287.1065, found: 287.1055.
Step C: methyl 2-[(6-chloro-5-methyl-pyridazin-3-yl)-[2-(2,2-dimethyl-l,3-dioxolan-4yl)ethyl]amino]thiazole-4-carboxylate
Using Nucleophile Substitution General Procedure starting from the product from Step B as starting material, 178 mg (35%) of the desired product was obtained.
*H NMR (500 MHz, DMSO-cA) 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-ESI (m/z): [M+H]+ calcd for Ci7H22ClN4O4S: 413.1045, found: 413.1048.
Step D: methyl 2-[[6-(l,3-benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-]2-(2,2dimethyl-l,3-dioxolan-4-yl)ethyl]amino]thiazole-4-carboxylate
320
Using Buchwald General Procedure II starting from the product from Step C as starting material in toluene at 150 °C for 4 h, 184 mg (81%) of the desired product was obtained.
HRMS-ESI (m/z): [M+H]+ calcd for C24H27N6O4S2: 527.1530, found 527.1512.
Step E: 2-[[6-(l,3-benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-(3,4dihydroxybutyl)amino]thiazole-4-carboxylic acid
To the solution of the product from Step D in 1,4-dioxane was added 2 M solution of NaOH and the mixture was stirred at 80 °C for 1 h. After acidifying the mixture with a 1 N solution of HCl, the reaction was stirred at 80 °C for 0.5 h, then the precipitate was filtered off and purified by flash column chromatography to give the desired product.
HRMS-ESI (m/z): [M+H]+ calcd for C20H21N6O4S2: 473.1066, found: 473.1055.
Example 86: 2-({6-[(l,3-Benzothiazol-2-yI)amino]-5-methylpyridazin-3yl}(methyl)ammo)-5-(l-benzylpiperidin-4-yI)-l,3-thiazoIe-4-carboxyIic acid
Step A: ethyl 5-(l-benzylpiperidin-4-yl)-2-[methyl(5-methyl-6-{[(2Z)-3-{[2(trimethylsilyl)ethoxy]methyl}-2,3-dihydro-l,3-benzothiazol-2-ylidene]amino}pyridazin-3yl)amino]-l,3-thiazole-4-carboxylate
To a stirred solution of the product from Préparation 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 pL) and the mixture was stirred at ambient température overnight. The reaction was partitioned between ethyl acetate and saturated aqueous sodium bicarbonate and the organic phase was washed with brine, dried (magnésium sulfate), and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 12 g RediSep™ silica cartridge) eluting with a gradient of 0 - 60% ethyl
321
A acetate in Ao-heptane afforded the desired product as a colourless gum (44.7 mg, 0.06 mmol, 60%).
LC/MS (C37H47N7O3SiS2) 730 [M+H]+; RT 1.32 (LCMS-V-B1)
Hl NMR (400 MHz, DMSO-d6) δ 7.89 (d, 1H), 7.66 (d, J = 1.1 Hz, 1H), 7.51 - 7.41 (m, 2H), 5 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).
Step B: ethyl 2-({6-[(l,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)10 5-(l-benzylpiperidin-4-yl)-l,3-thiazole-4-carboxylate
Hydrochloric acid (4M 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 was allowed to cool to ambient température, partitioned between dichloromethane and saturated aqueous sodium bicarbonate, and the 15 organic phase was washed with brine, dried (magnésium sulfate) and concentrated in vacuo.
Purification by automated flash column chromatography (CombiFlash Rf, 12 g RediSep™ silica cartridge) eluting with a gradient of 0 - 5% methanol in dichloromethane afforded the desired product as a yellow gum (22 mg, 0.04 mmol, 60%).
LC/MS (C3iH33N7O2S2) 600 [M+H]+; RT 0.99 (LCMS-V-B1) ’H NMR (400 MHz, DMSO-d6) 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-[(l,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)-5-(l25 benzylpiperidin-4-yl)-l,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 heated at reflux overnight, then allowed to cool to ambient température and concentrated in vacuo. The résultant solid was triturated with water then diethyl ether, filtered, and dried under vacuum to 30 afford the desired product as a yellow solid (9.6 mg, 0.02 mmol, 46%).
HRMS-ESI (m/z) [M+H]+ calcd for C29H30N7O2S2: 572.1897, found 572.1903.
322
Example 87: 2-({6-[(l,3-Benzothiazol-2-yl)amino]-5-methylpyridazin-3yl}(methyl)amino)-5-[l-(2-methylpropyl)piperidin-4-yl]-l,3-thiazole-4-carboxylic acid
Step A: ethyl 2-[methyl(5-methyl-6-{[(2Z)-3-{[2-(trimethylsilyl)ethoxy]methyl}-2,3-dihydrol,3-benzothiazol-2-ylidene]amino}pyridazin-3-yl)amino]-5-[l-(2-methylpropyl)piperidin-4yl]-l,3-thiazole-4-carboxylate
To a stirred solution of the product from Préparation 5k (134 mg, 0.21 mmol, 1 eq) in 2:1 acetonitrile / dichloromethane (6 mL) was added isobutyraldéhyde (0.04 mL, 0.42 mmol, 2 eq), sodium triacetoxyborohydride (133 mg, 0.63 mmol, 3 eq), and glacial acetic acid (10 pL) and the mixture wap stirred at ambient température overnight. The reaction mixture was partitioned between dichloromethane and saturated aqueous sodium bicarbonate, and the organic phase was washed with brine, dried (magnésium sulfate), and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 12 g RediSep™ silica cartridge) eluting with a gradient of 0 - 60% ethyl acetate in zso-heptane afforded the desired product as an off-whote solid (114 mg, 0.16 mmol, 79%).
LC/MS (C34H49N7O3SiS2) 696 [M+H]+; RT 1.30 (LCMS-V-B1)
Ή NMR (400 MHz, DMSO-d6) 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-[(l,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)5-[l-(2-methylpropyl)piperidin-4-yl]-l,3-thiazole-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 température overnight. The reaction was allowed to cool to ambient température
323
then partitioned between dichloromethane and saturated aqueous sodium bicarbonate, and the organic phase was washed with brine, dried (magnésium sulfate), and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 12 g RediSep™ silica cartridge) eluting with a gradient of 0 — 5% methanol in dichloromethane afforded the desired product as a yellow solid (76.8 mg, 0.14 mmol, 83%).
LC/MS (C28H35N7O2S2) 566 [M+H]+; RT 0.96 (LCMS-V-B1)
Hl NMR (400 MHz, DMSO-d6) 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, 10 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 Ç: 2-({6-[(l,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)-5-[l(2-methylpropyl)piperidin-4-yl]-l,3-thiazole-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 15 added lithium hydroxide monohydrate (28.5 mg, 0.68 mmol, 5 eq) and the mixture was heated at reflux overnight. The reaction was allowed to cool to ambient température and concentrated in vacuo. Purification by reverse phase automated flash chromatography (CombiFlash Rf, C18 13g RediSep column) eluting with a gradient of 5 - 95% acetonitrile in water afforded the desired product as a yellow solid (50 mg, 0.09 mmol, 69%).
HRMS-ESI (m/z) [M+H]+ calcd for C26H32N7O2S2: 538.2053, found 538.2060.
Example 88: 2-({6-[(l,3-Benzothiazol-2-yl)amino]-5-methylpyridazin-3yl}(methyl)amino)-5-[4-(phenoxymethyl)piperidin-l-yl]-Î,3-thiazole-4-carboxylic acid
324
Step A: ethyl 2-[methyl(5-methyl-6-{[(2Z)-3-{[2-(trimethylsilyl)ethoxy]methyl}-2,3-dihydrol,3-benzothiazol-2-ylidene]amino}pyridazin-3-yl)amino]-5-[4-(phenoxymethyl)piperidin-lyl]-l,3-thiazole-4-carboxylate
To a stirred solution of the product from Préparation 11b (200 mg, 0.31 mmol, 1 eq), 4(phenoxymethyl)piperidine hydrochloride (143 mg, 0.63 mmol, 2 eq) and césium carbonate (308 mg, 0.94 mmol, 3 eq) in 1,4-dioxane (8 mL) was added rac-BINAP-Pd-G3, (31.2 mg, 0.03 mmol, 0.1 eq) under a nitrogen atmosphère and the mixture was heated at 100 °C overnight. The reaction was allowed to cool to ambient température, then diluted with ethyl acetate, 10 washed with water then brine, dried (magnésium sulfate), and concentrated in vacuo to afford the desired product as a yellow foam (153 mg, 0.2 mmol, 65%).
LC/MS (C37H47N7O4SiS2) 746 [M+H]+; RT 1.60 (LCMS-V-B1) XH NMR (400 MHz, DMSO-d6) 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 = 15 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-[(l,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)5-[4-(phenoxymethyl)piperidin-l-yl]-l,3-thiazole-4-carboxylate
Hydrochloric acid (4M 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 was stirred at 50 °C overnight. The reaction was allowed to cool to ambient température, then partitioned between dichloromethane and saturated aqueous sodium bicarbonate, and the organic phase was washed with brine, dried (magnésium sulfate) and 25 concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash
I
325 ^b Rf, 12 g RediSep™ silica cartridge) eluting with a gradient of 0 - 5% methanol in dichloromethane afforded the desired product as a yellow glass (83.8 mg, 0.14 mmol, 67%).
LC/MS (C31H33N7O3S2) 616 [M+H]+; RT 1.39 (LCMS-V-B1)
Ή NMR (400 MHz, DMSO-d6) δ 11.01 (br s, 1H), 7.94 (br s, 1H), 7.63 (br s + s, 2H), 7.42 5 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-[(l,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)-5-[4(phenoxymethyl)piperidin-l-yl]-l ,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 heated at reflux overnight. The reaction was allowed to cool to ambient température and concentrated in vacuo. Purification by reverse phase automated flash chromatography (CombiFlash Rf, C18 13g RediSep column) eluting with a gradient of 5 - 95% acetonitrile in water afforded the 15 desired product as a yellow solid (59.7 mg, 0.1 mmol, 75%).
HRMS-ESI (m/z) [M+H]+ calcd for C29H30N7O3S2: 588.1846, found 588.1853.
Example 89: 5-(l-Benzoylpiperidin-4-yl)-2-({6-[(l,3-benzothiazol-2-yl)amino]-5methylpyridazm-3-yl}(methyl)amino)-l,3-thiazole-4-carboxylic acid
Step A: ethyl5-(l-benzoylpiperidin-4-yl)-2-[methyl(5-methyl-6-{[(2Z)-3-{[2(trimethylsilyl)ethoxy]methyl}-2,3-dihydro-l,3-benzothiazol-2-ylidene]amino}pyridazin-3yl)amino]-l,3-thiazole-4-carboxylate
To a stirred solution of the product from Préparation 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
I
326 chloride (0.02 mL, 0.17 mmol, 1.2 eq) and the mixture was stirred at ambient température for
4.5 h, then concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 12 g RediSep™ silica cartridge) eluting with a gradient of 0 - 100% ethyl acetate in Ao-heptane afforded the desired product as an off-white foam (89.6 mg, 0.12 mmol, 88%).
LC/MS (C37H45N7O4SiS2) 744 [M+H]+; RT 1.49 (LCMS-V-B1) *H NMR (400 MHz, DMSO-d6) 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-(l-benzoylpiperidin-4-yl)-2-({6-[(l,3-benzothiazol-2-yl)amino]-5methylpyridazin-3-yl}(methyl)amino)-l,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 (8,9.6 mg, 0.12 mmol, 1 eq) in dichloromethane (4 mL) and the mixture was stirred at ambient température overnight. The reaction was partitioned between dichloromethane and saturated aqueous sodium bicarbonate, and the organic phase was washed with brine, dried (magnésium sulfate) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 12 g RediSep™ silica cartridge) eluting with a gradient of 0 - 5% methanol in dichloromethane afforded the desired product as a yellow glass (55 mg, 0.09 mmol, 74%).
LC/MS (C3iH3iN7O3S2) 614 [M+H]+; RT 1.41 (LCMS-V-B1) 'H NMR (400 MHz, DMSO-d6) 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 (πί, 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 Ç: 5-(l-benzoylpiperidin-4-yl)-2-({6-[(l,3-benzothiazol-2-yl)amino]-5-methylpyridazin3-yl}(methyl)amino)-l,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 heated at reflux overnight. The reaction was allowed to cool to ambient température and concentrated
327 in vacuo. Purification by reverse phase automated flash chromatography (CombiFlash Rf, C18 13g RediSep column) eluting with a gradient of 5 - 95% acetonitrile in water afforded the desired product as a yellow solid (35.8 mg, 0.06 mmol, 68%).
HRMS-ESI (m/z) [M+H]+ calcd for C29H28N7O3S2: 586.1690, found 586.1702.
Example 90: 3-{l-[(Adamantan-l-yl)methyl]-5-methyl-lH-pyrazol-4-yl}-6-({6-[(l,3benzothiazol-2-yl)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
Césium carbonate (4.9 g, 15.1 mmol, 1.5 eq) was added to a solution of ethyl 3,6dibromopyridine-2-carboxylate (3.1 g, 10.03 mmol, 1 eq), ierFbutyl A-methylcarbamate (1.65 g, 12.6 mmol, 1.25 eq), tris(dibenzylideneacetone)dîpalladium(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) under a nitrogen atmosphère and the mixture was heated at 100 °C overnight. The reaction was allowed to cool to ambient température and ethyl acetate (250 mL) was added. The mixture was washed with water (2 x 100 mL), brine (100 mL), dried (magnésium sulfate) and concentrated in vacuo. Purification by flash column chromatography (100 g silica) eluting with dichloromethane afforded the desired product as a pale green oil (1.78 g, 4.96 mmol, 49%).
LC/MS (Ci4HÎ9BrN2O4) 303 [M-/Bu+H]+; RT 1.43 (LCMS-V.-B1) ‘H NMR (400 MHz, CDCI3) 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 to a solution of the product from Step A (1.78 g, 4.96 mmol, 1 eq) in dichloromethane (15 mL) at 0 °C and the mixture was allowed to warm to ambient température and stir overnight. The reaction was partitioned between
328 dichloromethane (50 mL) and saturated aqueous sodium bicarbonate, and the organic phase was washed with water (2 x 50 mL) and brine (50 mL), dried (magnésium sulfate), and concentrated in vacuo to afford the desired product as a yellow oil (1.2 g, 4.63 mmol, 94%).
LC/MS (C9HiiBrN2O2) 260 [M+H]+; RT 1.11 (LCMS-V-B1)
Hl NMR (400 MHz, CDC13) 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-l,2,4,5-tetrazin-3-yl)(methyl) amino]pyridine-2carboxylate
A solution of 3,6-dichloro-l,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 sealed flask for 18 h. The reaction was allowed to cool to ambient température and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 40 g RediSep™ silica cartridge) eluting with a gradient of 0 - 15% ethyl acetate in Ao-heptane afforded the desired product as a red solid (898 mg, 2.4 mmol, 55%).
I
LC/MS (CnHioBrCl N6O2) 373 [M+H]+; RT 1.31 (LCMS-V-B1) *H NMR (400 MHz, CDCI3) 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-2carboxylate
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 in a sealed tube at 100 °C for 24 h. The reaction was allowed to cool to ambient température and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 80 g RediSep™ silica cartridge) eluting with a gradient of 0 - 50% ethyl acetate in Aoheptane afforded the desired product as an orange solid (776 mg, 2.01 mmol, 85%).
LC/MS (Ci4Hi4BrClN4O2) 385 [M+H]+; RT 1.29 (LCMS-V-B1)
Ή NMR (400 MHz, CDCI3) 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).
329
Step E: ethyl 3-{!-[(adamantan-1 -yl)methyl]-5-methyl-lH-pyrazol-4-yl}-6-[(6-chloro-5methylpyridazin-3-yl)(methyl)amino]pyridine-2-carboxylate
To a suspension of the product from Step D (770 mg, 2 mmol, 1 eq) and the product from Préparation 10a (783 mg, 2.2 mmol, 1.1 eq) in tetrahydrofuran (12 mL) was added a solution 5 of potassium carbonate (552 mg, 3.99 mmol, 2 eq) in water (2 mL) and the mixture was sparged with nitrogen (10 mins). Pd(dppf)C12.CH2C12 (163 mg, 0.2 mmol, 0.1 eq) was added and the mixture was heated at reflux overnight. The reaction was allowed to cool to ambient température, then partitioned between ethyl acetate and brine and the organic phase was dried (magnésium Sulfate) and concentrated in vacuo. Purification by automated flash column 10 chromatography (CombiFlash Rf, 40 g RediSep™ silica cartridge) eluting with a gradient of 0 - 60% ethyl acetate in zso-heptane afforded the desired product as a cream solid (680 mg, 1.27 mmol, 64%).
LC/MS (C29H35CIN6O2) 535 [M+H]+; RT 1.55 (LCMS-V-B1) ΧΗ NMR (40Q MHz, DMSO-d6) δ 7.82 - 7.73 (m, 2H), 7.50 (d, J = 8.6 Hz, 1H), 7.35 (s, 1H), 15 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-{l-[(adamantan-1 -yl)methyl]-5-methyl-lH-pyrazol-4-yl}-6-({6-[(l,3benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)pyridine-2-carboxylate
To a solution of the product from Step E (600 mg, 1.12 mmol, 1 eq) and 2-aminobenzothiazole 20 (253 mg, 1.68 mmol, 1.5 eq) in 1,4-dioxane (20 mL) was added AjA-diisopropylethylamine (0.59 mL, 3.36 mmol, 3 eq), Xantphos (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 for 64 h. The reaction was allowed to cool to ambient température then diluted with ethyl acetate, washed with brine, and concentrated in vacuo. Purification by reverse 25 phase automated flash chromatography (CombiFlash Rf, C18 130g RediSep column) eluting with a gradient of 30-95% acetonitrile in water afforded the desired product as a yellow glass (513 mg, 0.79 mmol, 71%).
LC/MS (C36H40N8O2S) 649 [M+H]+; RT 1.62 (LCMS-V-B1)
330 φ ΧΗ NMR (400 MHz, DMSO-d6) 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-{l-[(adamantan-l-yl)methyl]-5-methyl-lH-pyrazol-4-yl}-6-({6-[(l,3-benzothiazol2-yl)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 heated at reflux for 1 h. The reaction was allowed to cool to ambient température and concentrated in vacuo then water (5 mL) was added and the pH was adjusted to 3 by the addition of 2M aqueous hydrochloric acid. The mixture was extracted into dichloromethane and the organic extract was washed with brine, dried (magnésium sulfate), and concentrated in vacuo. The résultant solid was triturated with methanol, filtered, washed with methanol, and dried under vacuum to afford the desired product as an off-white solid (362 mg, 0.58 mmol, 74%).
HRMS-ESI (m/z) [M+H]+ calcd for C34H37N8O2S: 621.2755, found 621.2765.
Example 91: 2-({6-[(l,3-Benzothiazol-2-yl)amino]-5-methylpyridazin-3yl}(methyl)amino)-5-(l-phenylpiperidin-4-yI)-l,3-thiazole-4-carboxylic acid
Step A: ethyl 2-[methyl(5-methyl-6-{[(2Z)-3-{[2-(trimethylsilyl)ethoxy]methyl}-2,3-dihydro20 l,3-benzothiazol-2-ylidene]amino}pyridazin-3-yl)amino]-5-(l-phenylpiperidin-4-yl)-l,3thiazole-4-carboxylate
Tris(dibenzylideneacetone)dipalladium(0) (10.7 mg, 0.01 mmol, 0.05 eq) was added to a stirred solution of th^ product from Préparation 5k (150 mg, 0.23 mmol, 1 eq), bromobenzene (0.04 mL, 0.35 mmol, 1.5 eq), césium carbonate (229 mg, 0.7 mmol, 3 eq) and Xantphos (13.6 mg, 25 0.02 mmol, 0.1 eq) in 1,4-dioxane (5 mL) under a nitrogen atmosphère and the mixture was
331 heated at 100 °C overnight. The reaction was allowed to cool to ambient température then partitioned between dichloromethane and water, and the organic phase was washed with water then brine, dried (magnésium sulfate), and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 12 g RediSep™ silica cartridge) eluting with a gradient of 0 - 50% ethyl acetate in Ao-heptane afforded the desired product as a brown foam (156 mg, 0.22 mmol, 93%).
LC/MS (C36H45N7O3SiS2) 716 [M+H]+; RT 1.68 (LCMS-V-B1) >H NMR (400 MHz, DMSO-d6) 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), -0.11 (s, 9H).
Step B: ethyl 2-({6-[(l,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)5-(l-phenylpiperidin-4-yl)-l,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 température overnight. The reaction was partitioned between dichloromethane and saturated aqueous sodium bicarbonate, and the organic phase was washed with brine, dried (magnésium sulfate), and concentrated in vacuo. The résultant solid was triturated with 1:1 dichloromethane / heptane, filtered, and dried under vacuum to afford the desired product as a yellow solid (92 mg, 0.16 mmol, 72%).
LC/MS (C3oH3iN702S2) 586 [M+H]+; RT 1.40 (LCMS-V-B1)
Hl NMR (400 MHz, DMSO-d6) 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-[(l,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)-5-(lphenylpiperidin-4-yl)-l,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 heated at reflux overnight. The reaction was allowed to cool to ambient température, and the solids collected by filtration, washed with 1,4-dioxane, then triturated in water, filtered and dried
332 under vacuum to afford the desired product as a yellow solid (50.3 mg, 0.09 mmol, 57%) [as a lithium sait].
HRMS-ESI (m/z) [M+H]+ calcd for C28H28N7O2S2: 558.1740, found 558.1749.
Example 92: 5-[l-(Benzenesulfonyl)piperidin-4-yl]-2-({6-[(l,3-benzothiazol-2-yl)amino]5 5-methyIpyridazin-3-yl}(methyl)amino)-l,3-thiazole-4-carboxylic acid
Step A: ethyl 5-[l-(benzenesulfonyl)piperidin-4-yl]-2-[methyl(5-methyl-6-{[(2Z)-3-{[2(trimethylsilyl)ethoxy]methyl}-2,3-dihydro-l,3-benzothiazol-2-ylidene]amino}pyridazin-3yl)amino]-l,3‘thiazole-4-carboxylate
To a stirred solution of the product from Préparation 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(dimethylamin!o)pyridine and the mixture was stirred at ambient température overnight. The reaction was partitioned between dichloromethane and water, and the organic phase was washed with brine, dried (magnésium sulfate), and concentrated in vacuo. Purification by automated fla^h column chromatography (CombiFlash Rf, 12 g RediSep™ silica cartridge) eluting with a gradient of 0 - 60% ethyl acetate in zso-heptane afforded the desired product as a cream solid (204 mg, 0.26 mmol, 84%).
LC/MS (C36H45N7O5SiS3) 780 [M+H]+; RT 1.53 (LCMS-V-B1) JH NMR (4ΟθΙ MHz, DMSO-d6) 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 5ç[l-(benzenesulfonyl)piperidin-4-yl]-2-({6-[(l,3-benzothiazol-2-yl)amino]-525 methylpyridazin-3-yl}(methyl)amino)-l,3-thiazole-4-carboxylate
333
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 température overnight. The reaction was partitioned between dichloromethane and saturated aqueous sodium bicarbonate, and the organic phase was washed with brine, dried (magnésium sulfate), and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 12 g RediSep™ silica cartridge) eluting with a gradient of 0 -5% methanol in dichloromethane afforded the desired product as a yellow glass (121 mg, 0.19 mmol, 71%).
LC/MS (C30H31N7O4S3) 650 [M+H]+; RT 1.28 (LCMS-V-B1)
Ή NMR (400 MHz, DMSO-d6) 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, 1H), 2.46 (s, 3H), 2.40 - 2.29 (m, 2H), 2.04 (d, J = 12.5 Hz, 2H), 1.75 ί 1.58 (m, 2H), 1.25 (t, J = 7.1 Hz, 3H).
Step C: 5-[l-(benzenesulfonyl)piperidin-4-yl]-2-({6-[(l,3-benzothiazol-2-yl)amino]-5methylpyridazin-3-yl}(methyl)amino)-l,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 heated at reflux overnight. The reaction was allowed to cool to ambient température and the solids collected by filtration and washed with 1,4 dioxane. The material was triturated with water, filtered, and dried under vacuum to afford the desired product as a yellow solid (102 mg, 0.16 mmol, 88%) [as a lithium sait].
HRMS-ESI (m/z) [M+H]+ calcd for C28H28N7O4S3: 622.1359, found 622.1367.
Example 93: 2-[[6-(l,3-Benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-(3-hydroxy-4methoxy-butyl)amino]thiazole-4-carboxylic acid
334
Step A: methyl2-[tert-butoxycarbonyl-[3-[tert-butyl(dimethyl)silyl]oxy-4-methoxybutyl]amino]thiazole-4-carboxylate
Using Mitsunobu General Procedure starting from methyl 2-(tertbutoxycarbonylamino)thiazole-4-carboxylate and Préparation 2h as starting materials in THF at rt for 1 h, 4.16 g (81%) of the desired product was obtained.
XH NMR (500 MHz, DMSO-cA) 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). 13C NMR (125 MHz, DMSO-dg) 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-methoxy-butyl]amino]thiazole-4carboxylate |
Using Deprotection with HFIP General Procedure starting from the product from Step A as the appropriate carbamate, 3.1 g (55%) of the desired product was obtained.
XH NMR (40C| MHz, DMSO-d6) 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-methoxy-butyl]-[5-methyl-6-[(Z)-[3-(2trimethylsilylethoxymethyl)-l,3-benzothiazol-2-ylidene]amino]pyridazin-3yl]amino]thiazole-4-carboxylate
Using Buchwald General Procedure II starting from the product from Step B and Préparation 4a as the appropriate halide, 118 mg (61%) of the desired product was obtained. Ή NMR (500 MHz, DMSO-î/6) 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); 13C NMR (125 MHz, DMSO-rfô) δ ppm 162.3, 160.4, 157.7, 155.3,150.9, 140.5, 137.5, 137.2, 127.2, 125.4, 123.5, 123.2, 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]+ calcd for ¢3^53^0582812: 745, found: 745.
StepD:2-[[6-(l,3-benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-(3-hydroxy-4-methoxybutyl)amino]thiazole-4-carboxylic acid
335
Using Deprotection and Hydrolysis General Procedure followed by repurification via reverse phase préparative chromatography (C18, 0.1% TFA in water : MeCN) starting from the product from Step C, TFA-salt of the desired product was obtained.
HRMS-ESI (m/z): [M+H]+ calcd for C21H23N6O4S2: 487.1217, found: 487.1222.
Example 94: 2-({6-[(l,3-Benzothiazol-2-yl)amino]-5-methylpyridazin-3yl}(methyl)amino)-5-(l-methanesulfonylpiperidin-4-yl)-l,3-thiazole-4-carboxylic acid
Step A: ethyl 5-(l-methanesulfonylpiperidin-4-yl)-2-[methyl(5-methyl-6-{[(2Z)-3-{[2(trimethylsilylethoxy]methyl}-2,3-dihydro-l,3-benzothiazol-2-ylidene]amino}pyridazin-3yl)amino]-l,3-thiazole-4-carboxylate
To a stirred solution of the product from Préparation 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) followed by a catalytic amount of 41 (dimethylamino)pyridine and the mixture was stirred at ambient température overnight. The reaction was diluted with dichloromethane then washed with water followed by brine, dried (magnésium sjulfate) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 12 g RediSep™ silica cartridge) eluting with a gradient of 0 - 60% ethyl acetate in Ao-heptane afforded the desired product as a white foam (106 mg, 0.15 mmol, 63%).
LC/MS (C3iH43N7O5SiS3) 718 [M+H]+; RT 1.45 (LCMS-V-B1) XH NMR (400 MHz, DMSO-d6) 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).
336
Step B: ethyl 2-({6-[(l,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)5-(l-methanesulfonylpiperidin-4-yl)-l,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 5 at ambient température overnight. The reaction was partitioned between dichloromethane and saturated aqueous sodium bicarbonate and the organic phase was washed with brine, dried (magnésium sulfate), and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 12 g RediSep™ silica cartridge) eluting with a gradient of 0 — 5% methanol in dichloromethane gave a solid that was triturated with diethyl ether, filtered, 10 and dried under vacuum to afford the desired product as a yellow solid (75 mg, 0.13 mmol,
86%). |
LC/MS (C25H29N7O4S3) 588 [M+H]+; RT 1.36 (LCMS-V-B1)
Ή NMR (400 MHz, DMSO-d6) 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), 15 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-[(l,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)-5-(lmethanesulfonylpiperidin-4-yl)-l,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 20 added lithium hydroxide monohydrate (26.8 mg, 0.64 mmol, 5 eq) and the mixture was heated at reflux overnight. The reaction was allowed to cool to ambient température and the solids collected by filtration and washed with 1,4 dioxane. Purification by reverse phase automated flash chromatography (CombiFlash Rf, C18 13g RediSep column) eluting with a gradient of 5 - 95% acetonitrile in water afforded the desired product as a yellow solid (37.1 mg, 0.07 mmol, 25 52%). |
HRMS-ESI (m/z) [M+H]+ calcd for C23H26N7O4S3: 560.1203, found 560.1209.
Example 95: 2-({6-[(l,3-Benzothiazol-2-yI)amino]-5-methylpyridazin-3yl}(methyl)amino)-5-(l-phenylmethanesulfonylpiperidin-4-yl)-l,3-thiazole-4-carboxylic acid
337
Step A: ethyl 2-[methyl(5-methyl-6-{[(2Z)-3-{[2-(trimethylsilyl)ethoxy]methyl}-2,3-dihydrol,3-benzothiazol-2-ylidene]amino}pyridazin-3-yl)amino]-5-(lphenylmethanesulfonylpiperidin-4-yl)-l,3-thiazole-4-carboxylate
To a stirred solution of the product from Préparation 5k and triethylamine (0.07 mL, 0.47 mmol, 2 eq) in dichloromethane (5 mL) was added α/ρ/ια-toluenesulfonyl chloride (53.6 mg, 0.28 mmol, 1.2 eq) followed by a catalytic amount of 4-(dimethylamino)pyridine and the mixture was stirred at àmbient température overnight. The reation was diluted with dichloromethane, washed with water followed by brine, dried (magnésium sulfate), and concentrated in vacuo.
Purification by automated flash column chromatography (CombiFlash Rf, 12 g RediSep™ silica cartridge) eluting with a gradient of 0 — 60% ethyl acetate in Ao-heptane afforded the desired product as a cream foam (95 mg, 0.12 mmol, 51%).
LC/MS (C37H47N7O5SiS3) 794 [M+H]+; RT 1.50 (LCMS-V-B1) 1H NMR (400 MHz, DMSO-d6) δ 7.81 (dd, J = 7.8,1.1 Hz, 1H), 7.67 (d, J = 1.1 Hz, 1H), 7.53 15 - 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-[(l,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)20 5-(l-phenylmethanesulfonylpiperidin-4-yl)-l,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 température overnight. The reaction was partitioned between dichloromethane and saturated aqueous sodium bicarbonate, and the organic phase was washed with brine, dried 25 (magnésium sulfate) and concentrated in vacuo. The résultant solid was triturated in diethyl i
338 ether, filtered, )and dried under vacuum to afford the desired product as a yellow solid (74 mg, 0.11 mmol, 93|%).
LC/MS (C31H33N7O4S3) 664 [M+H]+; RT 1.27 (LCMS-V-B1) XH NMR (400 MHz, DMSO-d6) δ 11.03 (br s, 1H), 7.91 (br s, 1H), 7.74 - 7.65 (m, 1H), 7.53 5 - 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), 2.50 - 2.44 (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-[(l,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)-5-(lphenylmethanesulfonylpiperidin-4-yl)-l,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 heated at reflux for 6.5 h. The reaction was allowed to cool to ambient température and purification by reverse phase automated flash chromatography (CombiFlash Rf, C18 13g RediSep column) eluting with a gradient of 5 — 95% acetonitrile in water afforded the desired product as a yellow 15 solid (43.2 mg; 0.07 mmol, 61%).
HRMS-ESI (m/z) [M+H]+ calcd for C29H3oN704S3: 636.1516, found 636.1521.
Example 96: 2-({6-[(l,3-Benzothiazol-2-yl)amino]-5-methylpyridazin-3yl}(methyI)amino)-5-[l-(2-phenylacetyl)piperidin-4-yl]-l,3-thiazole-4-carboxyIic acid
Step A: ethyl2-[methyl(5-methyl-6-{[(2Z)-3-{[2-(trimethylsilyl)ethoxy]methyl}-2,3-dihydrol,3-benzothiazol-2-ylidene]amino}pyridazin-3-yl)amino]-5-[l-(2-phenylacetyl)piperidin-4yl]-l,3-thiazole-4-carboxylate
To a stirred solution of the product from Préparation 5k (150 mg, 0.23 mmol, 1 eq) and triethylamine (0.07 mL, 0.47 mmol, 2 eq) in dichloromethane (5 mL) was added phenylacetyl
339 chloride (0.04 mL, 0.28 mmol, 1.2 eq) and the mixture was stirred at ambient température overnight. The reaction was diluted with dichloromethane then washed with saturated aqueous sodium bicarbonate followed by brine, dried (magnésium sulfate), and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 12 g RediSep™ silica cartridge) eluting with a gradient of 0 - 60% ethyl acetate in Ao-heptane afforded the desired product as an off-white foam (122 mg, 0.16 mmol, 69%).
LC/MS (C38hLn7O4SîS2) 758 [M+H]+; RT 1.49 (LCMS-V-B1) XH NMR (400 MHz, DMSO-d6) 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-[(l,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)5-[l-(2-phenylacetyl)piperidin-4-yl]-l,3-thiazole-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 température overnight. The reaction was partitioned between dichloromethane and saturated aqueous sodium bicarbonate, and the organic phase was washed with brine, dried (magnésium sulfate), and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 12 g RediSep™ silica cartridge) eluting with a gradient of 0 - 5% methano^ in dichloromethane afforded the desired product as a yellow solid (81.8 mg, 0.13 mmol, 81%).
LC/MS (C32H33N7O3S2) 628 [M+H]+; RT 1.22 (LCMS-V-B1) *H NMR (400 MHz, DMSO-d6) 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 Hzl, 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, ^H).
Step C: 2-({6-[(l,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)-5-[l(2-phenylacet^l)piperidin-4-yl]-l,3-thiazole-4-carboxylic acid
340
A To a solution of the product from Step B (81.8 mg, 0.13 mmol, 1 eq) in 1,4-fioxane (3 mL) was added lithium hydroxide monohydrate (27.3 mg, 0.65 mmol, 5 eq) and the mixture was heated at reflux overnight. The reaction was allowed to cool to ambient température and purification by reverse phase automated flash chromatography (CombiFlash Rf, C18 13g RediSep column) 5 eluting with a gradient of 5 - 95% acetonitrile in water afforded the desired product as a yellow solid (54.3 mg,, 0.09 mmol, 70%).
HRMS-ESI (m/z) [M+H]+ calcd for C30H30N7O3S2: 600.1846, found 600.1850.
Example 97: 5-(l-Acetylpiperidin-4-yl)-2-({6-[(l,3-benzothiazol-2-yl)amino]-5methylpyridazin-3-yl}(methyl)amino)-l,3-thiazole-4-carboxylic acid
Step A: ethyl5-(l-acetylpiperidin-4-yl)-2-[methyl(5-methyl-6-{[(2Z)-3-{[2(trimethylsilyl)ethoxy]methyl}-2,3-dihydro-l,3-benzothiazol-2-ylidene]amino}pyridazin-3yl)amino]-l,3-thiazole-4-carboxylate
To a stirred solution of the product from Préparation 5k and triethylamine (0.07 mL, 0.47 mmol, 15 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 température overnight. The reaction was diluted with dichloromethane then washed with saturated aqueous sodium bicarbonate followed by brine, dried (magnésium sulfate), and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 12 g RediSep™ silica cartridge) eluting with a gradient of 0 20 - 100% ethyl acetate in Lo-heptane afforded the desired product as a colourless foam (119 mg,
0.17 mmol, 74%).
LC/MS (C32H43N7O4S1S2) 682 [M+H]+; RT 1.44 (LCMS-V-B1) >H NMR (400 MHz, DMSO-d6) 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), 25 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 =
341
12.2 Hz, 1H), 2.69 - 2.54 (m, 1H), 2.45 (d, 3H), 2.06 (s, 3H), 2.06 - 1.95 (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 ï-(l-acetylpiperidin-4-yl)-2-({6-[(l,3-benzothiazol-2-yl)amino]-5methylpyridazin-3-yl}(methyl)amino)-l,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 (IM 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 was allowed to cool to ambient température, then partitioned between dichloromethane and saturated aqueous sodium bicarbonate, and the organic phase was washed with brine, dried (magnésium sulfate) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 12 g RediSep™ silica cartridge) eluting with a gradient of 0 - 4% methanol in dichloromethane afforded the desired product as a yellow solid (40 mg, 0.07 mmol, 42%).
LC/MS (C26H29N7O3S2) 552 [M+H]+; RT 1.09 (LCMS-V-B1)
Ή NMR (400 MHz, DMSO-d6) 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).
StepjÇ: 5-(l-acetylpiperidin-4-yl)-2-({6-[(l,3-benzothiazol-2-yl)amino]-5-methylpyridazin3-yl}(methyl)amino)-l,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 heated at reflux overnight. The reaction was allowed to cool to ambient température and purification by reverse phase automated flash chromatography (CombiFlash Rf, C18 13g RediSep column) eluting with a gradient of 5 - 95% acetonitrile in water afforded the desired product as a yellow solid (32.4 mg, 0.06 mmol, 85%).
HRMS-ESI (m/z) [M+H]+ calcd for C24H26N7O3S2·. 524.1533, found 524.1541.
Example 98:
2-[[6-(l,3-Benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-(4-hydroxy-330 methoxy-butyI)amino]thiazole-4-carboxylic acid
342
Step A : methyl 2-[tert-butoxycarbonyl-[4-[tert-butyl(dimethyl)silyl]oxy-3-methoxybutyl]amino]thiazole-4-carboxylate
Using Mitsunobu General Procedure starting from methyl 2-(tert5 butoxycarbonylamino)thiazole-4-carboxylate and Préparation 2i as starting materials in THF at rt for 3 h, 4.54 g (86%) of the desired product was obtained.
LC-MS-ESI (m/z): [M+H]+ calcd for C2iH39N2O6SSi: 475, found; 475.
Step B: methyl 2-[[4-[tert-butyl(dimethyl)silyl]oxy-3-methoxy-butyl]amino]thiazole-4carboxylate
Using Deprotection with HFIP General Procedure starting from the product from Step A as the appropriate carbamate, 2.88 g (55%) of the desired product was obtained.
Ή NMR (400 MHz, DMSO-·) 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-methoxy-butyl]-[5-methyl-6-[(Z)-[3-(215 trimethylsilylethoxymethyl)-l,3-benzothiazol-2-ylidene]amino]pyridazin-3yl]amino]thiaz.ple-4-carboxylate
Using Buchwald General Procedure II starting from the product from Step B and Préparation 4a as the appropriate halide, 319 mg (70%) of the desired product was obtained.
XH NMR (500 MHz, DMSO-·) δ ppm 7.99 (s, 1H), 7.85 (dm, 1H), 7.66 (s, 1H), 7.48 (dm, 20 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-(l,3-benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-(4-hydroxy-3-methoxybutyl)amino]thiazole-4-carboxylic acid
Using Deprotection and Hydrolysis General Procedure followed by repurification via
343 reverse phase préparative chromatography (C18, 0.1% TFA in water : MeCN) starting from the product from Step C, TFA-salt of the desired product was obtained.
HRMS-ESI (ψ/ζ): [M+H]+ calcd for C21H23N6O4S: 487.1217, found: 487.1222.
Example 99: 2-({6-[(l,3-Benzothiazol-2-yl)amino]-5-methylpyridazin-3yl}(methyl)amino)-5-(l-methylpiperidin-4-yl)-l,3-thiazole-4-carboxylic acid
Step A: ethyl2-[methyl(5-methyl-6-{[(2Z)-3-{[2-(trimethylsilyl)ethoxy]methyl}-2,3-dihydrol,3-benzothiazol-2-ylidene]amino}pyridazin-3-yl)amino]-5-(l-methylpiperidin-4-yl)-l,3thiazole-4-carboxylate
To a stirred solution of the product from Préparation 5k (150 mg, 0.23 mmol, 1 eq) in 1:1 acetonitrile / dichloromethane (10 mL) was added formaldéhyde (37% in water; 0.04 mL, 0.47 mmol, 2 eq), sodium triacetoxyborohydride (149 mg, 0.7 mmol, 3 eq), and glacial acetic acid (10 pL) and the mixture was stirred at ambient température overnight. The reaction was partitioned between dichloromethane and saturated aqueous sodium bicarbonate and the organic phase was washed with brine, dried (magnésium sulfate) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 12 g RediSep™ silica cartridge) eluting with a gradient of 0 — 100% ethyl acetate in zso-heptane afforded the desired producjt as an off-white foam (76.7 mg, 0.12 mmol, 50%).
LC/MS (C3iH43N7O3SiS2) 654 [M+H]+; RT 1.24 (LCMS-V-B1)
Hl NMR (400 MHz, DMSO-d6) 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).
344 k Step B: ethyl2-({6-[(l,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)5-(l-methylpiperidin-4-yl)-l,3-thiazole-4-carboxylate
Trifluoroaœtic 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 5 at ambient température overnight. The reaction was partitioned between dichloromethane and saturated aqueous sodium bicarbonate, and the organic phase was washed with brine, dried (magnésium sulfate), and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 12 g RediSep™ silica cartridge) eluting with a gradient of 0 - 10% methanol in dichloromethane afforded the desired product as a yellow glass (24.1 mg, 10 0.05 mmol, 39%).
LC/MS (C25H29N7O2S2) 524 [M+H]+; RT 0.87 (LCMS-V-B1) XH NMR (400 MHz, DMSO-d6) 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), 15 1.33 (t, J = 7.1 Hz, 3H).
Step C: 2-({6-[(l,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)-5-(lmethylpiperidin-4-yl)-l,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 heated 20 at reflux overnight. The reaction was allowed to cool to ambient température and concentrated in vacuo. Purification by reverse phase automated flash chromatography (CombiFlash Rf, C18 13g RediSep column) eluting with a gradient of 5 - 95% acetonitrile in water afforded the desired product as a yellow solid (17.2 mg, 0.03 mmol, 76%).
HRMS-ESI (ik/z) [M+H]+ calcd for C23H26N7O2S2: 496.1584, found 496.1602.
Example 100: 2-({6-[(l,3-Benzothiazol-2-yl)amino]-5-methylpyridazin-3yl}(methyl)amino)-5-{3-[2-fluoro-4-(3-hydroxyprop-l-yn-l-yl)phenoxy]propyl}-l,3thiazole-4-carboxylic acid
345
Step A: ethyl 5-[3-(2-fluoro-4-{3-[(4-methoxyphenyl)methoxy]prop-l-yn-lyl}phenoxy)propyl]-2-[methyl(5-methyl-6^[(2Z)-3-{[2-(trimethylsilyl)ethoxy]methyl}-2,3dihydro-l,3-benzothiazol-2-ylidene]amino}pyridazin-3-yl)amino]-l,3-thiazole-4-carboxylate
To a solution of the product from Préparation 5g (249 mg, 0.34 mmol, 1 eq) in dimethylformamide (5 mL) was added the product from Préparation 6k (128 mg, 0.45 mmol, 1.3 eq) and césium carbonate (336 mg, 1.03 mmol, 3 eq) and the mixture was heated at 80 °C overnight. The reaction was partitioned between ethyl acetate and water, and the organic phase was washed with brine, dried (magnésium sulfate), and concentrated in vacuo. Purification by 10 automated flash column chromatography (CombiFlash Rf, 12 g RediSep™ silica cartridge) eluting with a gradient of 0 - 50% ethyl acetate in Ao-heptane afforded the desired product as a yellow gum (231 mg, 0.26 mmol, 76%).
LC/MS (C45H5iFN6O6SiS2) 883 [M+H]+; RT 1.41 (LCMS-V-B2) XH NMR (400 MHz, DMSO-d6) δ 7.84 (d, 1H), 7.67 (d, J = 1.1 Hz, 1H), 7.49 - 7.40 (m, 2H), 15 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), .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-l-yn-1 -yl)phenoxy]propyl}-2-[methyl(520 methyl-6-{[(2Z)-3-{[2-(trimethylsilyl)ethoxy]methyl}-2,3-dihydro-l,3-benzothiazol-2ylidene]amind}pyridazin-3-yl)amino]-l,3-thiazole-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 température for 1 h. The reaction was diluted with dichloromethane, cooled to 0 °C, and neutralised by the 25 addition of 0.5M aqueous sodium hydroxide. The layers were separated and the organic phase
I
346 was dried (magnésium sulfate) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 12 g RediSep™ silica cartridge) eluting with a gradient of 0 - 50% ethyl acetate in Ao-heptane afforded the desired product as an off white solid (121 mg, 0.16 mmol, 36%).
LC/MS (C37H43FN6O5SiS2) 763 [M+H]+; RT 1.29 (LCMS-V-B2) XH NMR (400 MHz, DMSO-d6) 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-[(l,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)5-{3-[2-fluoro-4-(3-hydroxyprop-l-yn-l-yl)phenoxy]propyl}-l,3-thiazole-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 pL, 0.48 mmol, 3 eq) and TBAF (476 pL, 0.48 mmol, 3 eq) and the mixture was heated at 65 °C overnight. The reaction was allowed to cool to ambient température then partitioned between ethyl acetate and water, dried (magnésium sulfate), and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 12 g RediSep™ silica cartridge) eluting with a gradient of 0 - 100% ethyl acetate in Aoheptane afforded the desired product as a yellow solid (43 mg, 0.07 mmol, 43%).
LC/MS (C3iH29FN6O4S2) 633 [M+H]+; RT 2.43 (LCMS-V-C) XH NMR (400 MHz, DMSO-d6) 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-[(l,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)-5-{3[2-fluoro-4-(3-hydroxyprop-l-yn-1 -yl)phenoxy]propyl}-l,3-thiazole-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 heated at reflux overnight. The reaction was allowed to cool to ambient température and concentrated in vacuo. Purification by reverse phase automated flash chromatography (CombiFlash Rf, C18 13g RediSep column) eluting with a gradient of 30 - 95% acetonitrile in water afforded the desired product as a yellow solid (31.9 mg, 0.05 mmol, 67%).
34Ί
HRMS-ESI (m/z) [M-H]- calcd for C29H24FN6O4S2: 603.1290, found 603.1294.
Example 101:3-[3-[4-[3-[2-[[6-(l,3-Benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]methyI-amino]-4-carboxy-thiazol-5-yl]propoxy]-3-fluoro-phenyl]prop-2-ynyl-dimethylammonio] propane-l-sulfonate
Using Alkylation, Deprotection and Hydrolysis General procedure starting from Example , Step A and oxathiolane 2,2-dioxide, the desired product was obtained.
HRMS-ESI (m/z): [M+H]+ calcd for C34H37FN7O6S3: 754.1946, found: 754.1947.
Example 102: 2-({6-[(l,3-Benzothiazol-2-yl)amino]-5-methylpyridazin-3 yl}(methyl)amino)-5-cyclohexyl-l,3-thiazole-4-carboxylic acid
Step 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 under high vacuum then allowed to cool to ambient température and placed under nitrogen on a 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. lodocyclohexane (2 mL, 15.4 mmol, 1
348 eq) was added slowly then the mixture heated at 40 °C overnight. The reaction was allowed to cool to ambient température and cannulation through a filter (cotton-wool/ celite / cotton-wool) under slight vacuum into a dry 25 mL Schlenk tube afforded the desired product as a IM solution (as determined by titration with a 0.5M solution of iodine) that was used without further 5 characterisation.
Step B: ethyl 5-cyclohexyl-2-[methyl(5-methyl-6-{[(2Z)-3-{[2-(trimethylsilyl)ethoxy]methyl}2,3-dihydro-l,^-benzothiazol-2-ylidene]amino}pyridazin-3-yl)amino]-l,3-thiazole-4carboxylate |
To an oven-dried flask was added a solution of the product from Préparation 11b (200 mg, 0.31 10 mmol, 1 eq) and copper (I) iodide (112 mg, 0.63 mmol, 2 eq). Dimethylacetamide (5 mL) was added and the mixture was stirred for 5 min before the product from Step A (IM in dimethylacetamide; 1.42 mL, 1.42 mmol, 4.5 eq) was added slowly and the mixture was stirred at ambient température overnight. The reaction mixture was poured onto saturated aqueous ammonium chloride then extracted with dichloromethane (x2). The combined organic extracts 15 were washed with brine, dried (magnésium sulfate), and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 12 g RediSep™ silica cartridge) eluting with a gradient of 0 - 30% ethyl acetate in zso-heptane afforded the desired product as a yellow gum (135 mg, 0.21 mmol, 67%).
LC/MS (C3iH42N6O3S2Si) 639 [M+H]+; RT 1.68 (LCMS-V-B1) XH NMR (400|MHz, DMSO-d6) δ 7.85 (dd, J = 7.6, 1.0 Hz, 1 H), 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)i 1.52 - 1.35 (m, 4H), 1.32 (t, 3H), 0.96 - 0.88 (m, 2H), -0.11 (s, 9H).
Step C: 2-({6-[(l,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)-525 cyclohexyl-l,3-thiazole-4-carboxylic acid
Trifluoroacetic acid (1 mL, 13.4 mmol, 63.7 eq) was added to a stirred solution of the product from Step B (135 mg, 0.21 mmol, 1 eq) in dichloromethane (6 mL) and the mixture was stirred at ambient température overnight. The reaction was partitioned between dichloromethane and saturated aqueous sodium bicarbonate, and the organic phase was washed with brine, dried 30 (magnésium sulfate), and concentrated in vacuo. The résultant solid was suspended in 1,4dioxane (4 mL) then lithium hydroxide (44.2 mg, 1.05 mmol, 5 eq) added and the mixture was heated at reflux overnight. The reaction was allowed to cool to ambient température and
349 concentrated in vacuo. Purification by reverse phase automated flash chromatography (CombiFlash Rf, C18 13g RediSep column) eluting with a gradient of 5 - 95% acetonitrile in water afforded the desired product as a yellow solid (42 mg, 0.09 mmol, 41%).
HRMS-ESI (m/z) [M+H]+ calcd for C23H25N6O2S2: 481.1475, found 481.1518.
Example 103:2-[ï6-(1.3-Benzothiazol-2-vlamino)-5-methvl-pvridazin-3-vll-(2,3dihydroxypropyl)amino]thiazole-4-carboxylic acid
HO
Step A: methyl 2-[tert-butoxycarbonyl~[(2,2-dimethyl-l,3-dioxolan-4yl)methyl]amino]thiazole-4-carboxylate
Using Mitsunobu General Procedure starting from methyl 2-(tertbutoxycarbonylamino)thiazole-4-carboxylate and (2,2-dzmet/iy/-7,5-i/zoxoZ«n-4-yZ)mei/z«/io/ as starting materials in THF at rt for 19 h, 5.09 g (91%) of the desired product was obtained.
Ή NMR (500 MHz, DMSO-ίΑ) 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); 13C NMR (125 15 MHz, DMSO-i/e) 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; HRMS-ESI (m/z): [M+H]+ calcd for C16H25N2O6S: 373.1433 found: 373.1431.
Step B: methyl 2-[(2,2-dimethyl-l,3-dioxolan-4-yl)methylamino]thiazole-4-carboxylate
Using Deprotection with HFIP General Procedure starting from the product from Step A as the appropriate carbamate, 1.77 g (48%) of the desired product was obtained.
Ή NMR (500 MHz, DMSO-d6) 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); 13C NMR (125 MHz, DMSO-dô) δ ppm 168.7, 162.0, 142.3, 117.6, 74.3, 67.0, 52.1, 47.4, 27.3, 25.8; HRMS-ESI (m/z): [M+H]+ calcd for C11H17N2O4S: 273.0909 found: 273.0906.
Step C: methyl 2-[(2,2-dimethyl-l,3-dioxolan-4-yl)methyl-[5-methyl-6-[(Z)-[3-(2I
350 trimethylsilylethoxymethyl)-l,3-benzothiazol-2-ylidene]amino]pyridazin-3yl]amino]thiazole-4-carboxylate
Using Buchwald General Procedure II starting from the product from Step B and Préparation 4a as the appropriate halide, 450 mg (33%) of the desired product was obtained.
Ή NMR (500 MHz, DMSO-rfc) 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); 13C NMR (125 MHz, DMSO-rfc) 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; HRMS-ESI (m/z): [M+H]+ calcd for C29H39N6O5S2S1: 643.2192 found: 643.2184.
Step D: 2-[[6-(l,3-benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-(2,3dihydroxypropyl)amino]thiazole-4-carboxylic acid
Using Deprotection and Hydrolysis General Procedure followed by repurification via reverse phase préparative chromatography (C18, 25 mM NH4HCO3 water : MeCN) starting from the product from Step C, the desired product was obtained.
HRMS-ESI (m/z): [M+H]+ calcd for C19H19N6O4S2: 459.0909 found: 459.0891.
Example 104:2-[[6-(l,3-Benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-(4,5dihydroxypentyl)amino] thiazole-4-carboxylic acid
Step A: methyl2-[tert-butoxycarbonyl(pent-4-enyl)amino]thiazole-4-carboxylate
Using Mitsunobu General Procedure starting from methyl 2-(tertbutoxycarbonylamino)thiazole-4-carboxylate and 4-penten-l-ol as starting materials in THF at rt for 2 h, 4.36 g (89%) of the desired product was obtained.
Step B: methyl 2-[3-(2,2-dimethyl-l,3-dioxolan-4-yl)propylamino]thiazole-4-carboxylate
351
Το 4.35 g of the product from Step A (13.3 mmol) was added 3.6 g of 4-methyl-4-oxidomorpholin-4-ium;hydrate (26.6 mmol, 2 eq.), 135 mg of tetraoxoosmium (2.5 w% in 2methylpropan-2-ol, 0.013 mmol, 0.001 eq.), 10 mL of water, and 100 mL of 2-methylpropan2-ol and the mixture was stirred at rt for 18 h. After the addition of 50 mL of 1 M Na2S3O3 and removal of the lBuOH under reduced pressure, the mixture was extracted with DCM and the combined organic phases were dried and concentrated to give 6.25 g (130%) of methyl 2-[tertbutoxycarbonyl(4,5-dihydroxypentyl)amino]thiazole-4-carboxylate. The crude product was taken up in 100 mL of EtOH, treated with 20 mL of a 1 N HCl at 100 °C for 24 h, and concentrated to give 5.42 g (157%) of methyl 2-(4,5-dihydroxypentylamino)thiazole-4carboxylate (LC-MS-ESI (m/z): [M+H]+ calcd for C10H17N2O4S: 261, found 261). After the product was taken up in 50 mL of 2,2-dimethoxypropane and the addition of 124 mg of 4methylbenzenesulfonic acid (0.72 mmol, 0.05 eq.), the mixture was stirred at reflux for 3 h. After concentration, the residue was taken up in DCM and washed with cc. NaHCO3 to give 3.66 g (92% fdr three steps) of the the desired product.
Ή NMR (500 MHz, DMSO-î/6) 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); 13C NMR (125 MHz, DMSO-d6) δ ppm 117.0, 75.6, 69.0, 52.0, 44.7, 27.3, 27.3, 26.1, 25.5; HRMS-ESI (m/z): [M+H]+ calcd for Ci3H2iN2O4S: 301.1222 found: 301.1217.
Step C: methyl 2-[3-(2,2-dimethyl-l,3-dioxolan-4-yl)propyl-[5-methyl-6-[(Z)-[3-(2trimethylsilylethoxymethyl)-l,3-benzothiazol-2-ylidene]amino]pyridazin-3yl]amino]thiazole-4-carboxylate
Using Buchwald General Procedure II starting from the product from Step B and Préparation 4a as the appropriate halide, 74 mg (50%) of the desired product was obtained.
>H NMR (500 MHz, DMSO-d6) 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); 13C NMR (125 MHz, DMSO-ifo) 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; HRMS-ESI (m/z): [M+H]+ calcd for C3iH43N6O5S2Si: 671.2505 found: 671.2489.
Step D: 2-[[6-(l,3-benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-(4,5dihydroxypentyl)amino]thiazole-4-carboxylic acid
I 352
Using Deprotection and Hydrolysis General Procedure followed by repurification via reverse phase préparative chromatography (C18, 25 mM NH4HCO3 water : MeCN) starting from the product from Step C, the desired product was obtained.
HRMS-ESI (m/z): [M+H]+ calcd for C2iH23N6O4S2: 487.1222, found: 487.f 212.
Example 105: 2-({6-[(l,3-Benzothiazol-2-yl)amino]-5-methylpyridazin-3yl}(methyl)amino)-5-cyclopentyl-l,3-thiazoIe-4-carboxylic acid
Step A: bromo^cyclopentyl)zinc
Zinc (1.83 g, 2β mmol, 3 eq) and lithium chloride (593 mg, 14 mmol, 1.5 eq) were added to an 10 oven-dried Schlenk flask (50 mL). The flask was placed under vacuum and heated at 160 °C for 20 min, then allowed to cool to ambient température and placed under nitrogen on a 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 then the reaction heated at 40 °C overnight. The reaction mixture was 15 allowed to cool to ambient température and cannulation through a filter (cotton-wool/ celite / cotton-wool) uLder slight vacuum into a dry 25 mL Schlenk tube afforded the desired product as a 0.25M solution (as determined by titration with a 0.5M solution of iodine) that was used without further characterisation.
Step B: ethyl 5-cyclopentyl-2-[methyl(5-methyl-6-{[(2Z)-3-{[220 (trimethylsilyl)ethoxy]methyl}-2,3-dihydro-l,3-benzothiazol-2-ylidene]amino}pyridazin-3yl)amino]-l,3-thiazole-4-carboxylate
To an oven-dried flask was added the product from Préparation 11b (200 mg, 0.31 mmol, 1 eq) and copper (I) iodide (120 mg, 0.63 mmol, 2 eq) followed by dimethylacetamide (5 mL) and the mixture was stirred for 5 min before the product from Step A (0.25M in dimethylacetamide; 25 5.66 mL, 1.42 mmol, 4.5 eq) was added slowly and the mixture was stirred overnight. The reaction was poured onto saturated aqueous ammonium chloride and extracted with
353 dichloromethane (x2). The organic phase was washed with brine, dried (magnésium sulfate), and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 12 g RediSep™ silica cartridge) eluting with a gradient of 0 - 50% ethyl acetate in z'so-heptane afforded a solid that wastriturated with zso-heptane, filtered, washed with diethyl ether, and dried under vacuum to afford the desired product as a white solid (112 mg, 0.18 mmol, 57%).
LC/MS (C3oH4oN603S2Si) 625 [M+H]+; RT 1.79 (LCMS-V-B1)
Tl NMR (400 MHz, DMSO-d6) 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-[(l,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)-5cyclopentyl-l,3-thiazole-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 température overnight. The reaction was partitioned between dichloromethane and saturated aqueous sodium bicarbonate, and the organic phase was washed with brine, dried (magnésium sulfate), and concentrated in vacuo. The résultant solid was suspended in 1,4dioxane (3 mL) then lithium hydroxide (37.6 mg, 0.9 mmol, 5 eq) was added and the mixture was heated at reflux overnight. The reaction was allowed to cool to ambient température and concentrated in vacuo. Purification by reverse phase automated flash chromatography (CombiFlash Rf, C18 13g RediSep column) eluting with a gradient of 5 - 95% acetonitrile in water afforded the desired product as a brown solid (34.8 mg, 0.07 mmol, 42%).
HRMS-ESI (m/z) [M-H]- calcd for C22H21N6O2S2: 465.1173, found 465.1175.
Example 106: 2-({6-[(l,3-Benzothiazol-2-yl)amino]-5-methylpyridazin-3yl}(methyl)amino)-5-ethenyl-l,3-thiazole-4-carboxylic acid
354
Step A: ethyl5-ethenyl-2-[methyl(5-methyl-6-{[(2Z)-3-{[2-(trimethylsilyl)ethoxy]methyl}2,3-dihydro-l,3-benzothiazol-2-ylidene]amino}pyridazin-3-yl)amino]-l,3-thiazole-4carboxylate
To a solution of the product from Préparation 11b (100 mg, 0.16 mmol, 1 eq) in 5:1 tetrahydrofuran / water (6 mL) was added Pd(dppf)C12.CH2C12 (12.9 mg, 0.02 mmol, 0.1 eq), potassium vinyltrifluoroborate (31.6 mg, 0.24 mmol, 1.5 eq) and potassium carbonate (65.2 mg, 0.47 mmol, 3 eq) and the mixture was heated at reflux for 3 days. The reaction was allowed to cool to ambient température, then partitioned between ethyl acetate and water, and the organic 10 phase was washed with brine, dried (magnésium sulfate), and concentrated in vacuo.
Purification by automated flash column chromatography (CombiFlash Rf, 12 g RediSep™ silica cartridge) eluting with a gradient of 0 - 50% ethyl acetate in Zso-heptane afforded the desired produit as a white solid (39.5 mg, 0.07 mmol, 43%).
LC/MS (C27H34N6O3S2Si) 583 [M+H]+; RT 1.52 (LCMS-V-B1) XH NMR (400 MHz, DMSO-d6) 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-['(l,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)-520 ethenyl-l,3-thiazole-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 température overnight. The reaction was partitioned between dichloromethane and saturated aqueous sodium bicarbonate, and the organic phase was washed with brine, dried 25 (magnésium sulfate), and concentrated in vacuo. The résultant solid was suspended in 1,4dioxane (3 mL), lithium hydroxide monohydrate (14.2 mg, 0.34 mmol, 5 eq) was added, and the mixture was heated at reflux overnight. The reaction was allowed to cool to ambient
355 température and concentrated in vacuo. Purification by reverse phase automated flash chromatography (CombiFlash Rf, C18 13g RediSep column) eluting with a gradient of 5 - 95% acetonitrile m water afforded the desired product as a yellow solid (13.4 mg, 0.03 mmol, 47%).
HRMS-ESI (m/z) [M-H]- calcd for C19H15N6O2S2: 423.0703, found 423.0711.
Example 107:2-rr6-(1.3-Benzothiazol-2-vlamino)-5-methyl-pyridazin-3-yl]-(3-hydroxy-2morpholino-propyl)amino]thiazole-4-carboxylic acid
Step A: methyl 2-[tert-butoxycarbonyl-[3-[tert-butyl(dimethyl)silyl]oxy-2-morpholinopropyl]amino]thiazole-4-carboxylate
Using Mitsunobu General Procedure starting from methyl 2-(tertbutoxycarbonylamino)thiazole-4-carboxylate and Préparation 2k as starting materials at rt for 1 h, 1.29 g (89%) of the desired product was obtained.
Ή NMR (500 MHz, DMSO-d6) 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); 13C NMR (125 MHz, DMSO-d6) 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; HRMSESI (m/z): [M-Lh]+ calcd for C23H43N3O6SS1: 516.2564, found: 516.2570.
Step B: methyl 2-[[3-[tert-butyl(dimethyl)silyl]oxy-2-morpholino-propyl]amino]thiazole-4carboxylate
Using Deprotection with HFIP General Procedure starting from the product from Step A as the appropriate carbamate, 0.71 g (68%) of the desired product was obtained.
*H NMR (500 MHz, DMSO-d6) δ ppm 7.58 (t, 1H), 7.51 (s, 1H), 3.74/3.66 (dd+dd, 2H), 3.73 (s, 3H), 3.59-3.45 (m, 4H), 3.32/3.29 (m+m, 2H), 2.71 (m, 1H), 2.70/2.56 (m+m, 4H), 0.87 (s,
I 356 φ 9Η), 0.04 (s, 1h); 13C NMR (125 MHz, DMSO-ifc) 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; HRMS-ESI (m/z): [M-C4H9]+ calcd for Ci4H24N3O4SSi: 358.1262, found: 358.1255.
Step C: methyl 2-[[3-[tert-butyl(dimethyl)silyl]oxy-2-morpholino-propyl]-[5-methyl-6-[(Z)5 [3-(2-trimethylsilylethoxymethyl)-l,3-benzothiazol-2-ylidene]amino]pyridazin-3yl]amino]thia^ole-4-carboxylate
Using Buchwald General Procedure II starting from the product from Step B and Préparation 4a as the appropriate halide followed by purification via reverse phase préparative chromatography (C18, 25 mM NH4HCO3 water : MeCN), 270 mg (70%) of the desired product was obtained.
1H NMR (500 MHz, DMSO-rfc) 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, 9|H), 0.06 (s, 6H), -0.12 (s, 9H); 13C NMR (125 MHz, DMSO-î/6) δ ppm 127.2, 15 123.5,123.2,122.9,118.5,112.0, 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; HRMS-ESI (m/z): [M+H]+ calcd for C36H56N7O5S2Si2: 786.3322, found: 786.3312.
Step D: 2-[[6-(l,3-benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-(3-hydroxy-2morpholino-propyl)amino]thiazole-4-carboxylic acid
Using Deprotection and Hydrolysis General Procedure followed by repurification via 20 reverse phase préparative chromatography (C18, 25 mM NH4HCO3 water : MeCN) starting from the product from Step C, the desired product was obtained.
HRMS-ESI (m/z): [M+H]+ calcd for C23H26N7O4S2: 528.1487, found: 528.1489.
Example 108:2-ir6-(l,3-Benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-(3,4dihydroxybutyl)amino]-5-[3-[2-fluoro-4-[3-(methylamino)prop-l25 ynyl]phenoxy]propyl]thiazole-4-carboxylic acid
I
Step A: methyl5-[3-[4-[3-[tert-butoxycarbonyl(methyl) amino]prop-l-ynyl]-2-fluorophenoxy]propyl]-2-[2-(2,2-dimethyl-l,3-dioxolan-4-yl)ethyl-[5-methyl-6-[(Z)-[3-(2trimethylsilylethoxymethyl)-l,3-benzothiazol-2-ylidene]amino]pyridazin-3- yl]amino]thiazole-4-carboxylate
Using Buchwald General Procedure II starting from 350 mg of Préparation 3h (0.57 mmol, 1 eq.) and 235 mg of Préparation 4a (0.57 mmol, 1 eq.) as the appropriate halide, 490 mg (87%) of the desired product was obtained.
>H NMR (500 MHz, DMSO-d6) δ ppm 7.84 (d, 1H), 7.68 (s, 1H), 7.47 (d, 1H), 7.44 (td, 1H), 10 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), (1.17 (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); 13C NMR (125 MHz, DMSO-d6) 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, 15 30.9, 28.5, 27.3/26.0, 23.3, 23.1, 17.9, 17.8, -1.0; HRMS-ESI (m/z): [M+H]+ calcd for
C48H63FN7O8S2S1: 976.3927, found 976.3916.
Step B: 2-[[6-(l,3-benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-(3,4dihydroxybutyl)amino]-5-[3-[2-fluoro-4-[3-(methylamino)prop-lynyl]phenoxy]propyl]thiazole-4-carboxylic acid I
Using Deprotection and Hydrolysis General Procedure starting from the product from Step A as the appropriate methyl ester, the desired product was obtained.
HRMS-ESI (m/z): [M+H]+ calcd for C33H35FN7O5S2: 692.2120, found 692.2114.
358
Example 109^2-ri6-(1.3-Benzothiazol-2-vlamino)-5-methvl-pyridazin-3-yll-(3-hydroxy-4methoxy-butyl)amino]-5-[3-[2-fluoro-4-[3-(methylamino)prop-lynyl] phenoxy] propyl] thiazole-4-carboxylic acid
Step A: methyl 5-[3-[4-[3-[tert-butoxycarbonyl(methyl)amino]prop-l-ynyl]-2-fluorophenoxy]propyl]-2-[[3-[tert-butyl(dimethyl)silyl]oxy-4-methoxy-butyl]-[5-methyl-6-[(Z)-[3(2-trimethylsilylethoxymethyl)-l,3-benzothiazol-2-ylidene]amino]pyridazin-3yl]amino]thiazole-4-carboxylate
Using Buchwald General Procedure II starting from 320 mg of Préparation 31 (0.46 mmol, 10 1 eq.) and 188 mg of Préparation 4a (0.46 mmol, 1 eq.) as the appropriate halide, 415 mg (84%) of the desired product was obtained.
Ή NMR (50Q MHz, DMSO-d6) 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 (si, 9H), 0.08/0.06 (s/s, 6H), -0.12 (s, 9H); 13C NMR (125 MHz, DMSO-d6) δ ppm
I
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, 30.9, 28.5, 26.3, 23.1, 17.8, 17.8, -1.0, -4.1/-4.5; HRMS-ESI (m/z): [M+H]+ calcd for C52H75FN7O8S2Si2: 1064.4636, found 1064.4629.
Step B: 2-[[6-(l,3-benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-(3-hydroxy-4-methoxybutyl)amino]-5-[3-[2-fluoro-4-[3-(methylamino)prop-l-ynyl]phenoxy]propyl]thiazole-4carboxylic acid
Using Deprotection and Hydrolysis General Procedure starting from the product from Step A as the appropriate methyl ester, the desired product was obtained.
HRMS-ESI (m/z): [M+H]+ calcd for C34H37FN7O5S2: 706.2276, found 706.2285.
359
Example 110:2-[r6-(1.3-Benzothiazol-2-vlamino)-5-methvl-pyridazin-3-vll-(4hydroxybutyl)amino]thiazole-4-carboxylic acid
Step A: methyl 2-[tert-butoxycarbonyl-[4-[tert-butyl(dimethyl)silyl]oxybutyl]amino]thiazole5 4-carboxylate
Using Mitsunobu General Procedure starting from methyl 2-(tertbutoxycarbonylamino)thiazole-4-carboxylate and 4-[tert-butyl(dimethyl)silyl]oxybutan-l-ol as starting materials in THF at rt for 1 h, 3.38 g (98%) of the desired product was obtained.
*H NMR (500 MHz, DMSO-î/6) δ ppm 8.09 (s, 1H), 4.07 (t, 2H), 3.79 (s, 3H), 3.59 (t, 2H), 10 1.69 (qn, 2H)„ 1.53 (s, 9H), 1.45 (qn, 2H), 0.83 (s, 9H), 0.00 (s, 6H); 13C NMR (125 MHz,
DMSO-î/6) 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; HRMS-ESI (m/z): [M+H]+ calcd for CzoHstNzOsSSî: 445.2192 found: 445.2193.
Step B: methyl 2-[tert-butoxycarbonyl-[4-[tert-butyl(dimethyl)silyl]oxybutyl]amino]thiazole4-carboxylate
Using Deprotection with HFIP General Procedure starting from the product from Step A as the appropriate carbamate, 2.0 g (80%) of the desired product was obtained.
*H NMR (500 MHz, DMSO4) 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); 13C NMR (125 MHz, DMSO-dô) 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, 20 1732; HRMS-ESI (m/z): [M+H]+ calcd for Ci5H29N2O3SSi: 345.1668 found: 345.1652.
Step C: methyl 2-[4-[tert-butyl(dimethyl)silyl]oxybutyl-[5-methyl-6-[(Z)-[3-(2trimethylsilylethoxymethyl)-l,3-benzothiazol-2-ylidene]amino]pyridazin-3yl]amino]thiazole-4-carboxylate
Using Buchwald General Procedure II starting from the product from Step B and 25 Préparation 4a as the appropriate halide, 451 mg (73%) of the desired product was obtained.
360 XH 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); 13C NMR (125 MHz, DMSO-î/6) 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; HRMS-ESI (m/z): [M+H]+ calcd for C33H5iN6O4S2Si2: 715.2952 found: 715.2963.
Step D: 2-[[6-(l,3-benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-(4hydroxybutyl)amino]thiazole-4-carboxylic acid
Using Deprotection and Hydrolysis General Procedure followed by repurification via reverse phase préparative chromatography (C18, 25 mM NH4HCO3 water : MeCN) starting from the product from Step C, the desired product was obtained.
HRMS-ESI (m/z): [M+H]+ calcd for C20H21N6O3S2: 457.1117, found: 457.1108.
Example lll:2-ri6-(1.3-Benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-(3-hydroxy-2methoxy-propyl)amino]thiazole-4-carboxylic acid
Step A: methyl 2-[tert-butoxycarbonyl-[3-[tert-butyl(dimethyl)silyl]oxy-2-methoxypropyl]amino]thiazole-4-carboxylate
Using Mitsunobu General Procedure starting from methyl 2-(tertbutoxycarbonylamino)thiazole-4-carboxylate and Préparation 2j as starting materials in THF at rt for 1 h, 683 mg (88%) of the desired product was obtained.
Ή NMR (400 MHz, DMSO-ώ) 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)silylJoxy-2-methoxy-propyl]amino]thiazole-4carboxylate
361
Using Deprotection with HFIP General Procedure starting from the product from Step A as the appropriate carbamate, 514 mg (quant.) of the desired product was obtained.
'H NMR (400 MHz, DMSO-·) 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]+ calcd for Ci5H29N2O4SSi: 361, found: 361.
Step C: methyl 2-[[3-[tert-butyl(dimethyl)silyl]oxy-2-methoxy-propyl]-[5-methyl-6-[(Z)-[3(2-trimethylsilylethoxymethyl)-l,3-benzothiazol-2-ylidene]amino]pyridazin-3yl]amino]thiazole-4-carboxylate
I
Using Buchwald General Procedure II starting from the product from Step B and Préparation 4a as the appropriate halide, 313 mg (70%) of the desired product was obtained.
1H NMR (500 MHz, DMSO-·) 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-(l,3-benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-(3-hydroxy-2-methoxypropyl)amino]thiazole-4-carboxylic acid
Using Deprotection and Hydrolysis General Procedure followed by repurification via reverse phase préparative chromatography (C18, 0.1% TFA in water : MeCN) starting from the product from Step C, TFA-salt of the desired product was obtained.
HRMS-ESI (m/z): [M+H]+ calcd for C20H2iN6O4S2: 473.1066 found: 473.1063.
Example 112:2-iï6-(l.3-Benzothiazol-2-vlamino)-5-methvl-pyridazin-3-yl1-(5-hvdroxv-4methoxy-pentyl)amino]-5-[3-[2-fluoro-4-[3-(methylamino)prop-lyny 1] phenoxy] propyl] thiazole-4-carboxylie acid
362
Step A: methyl5-[3-[4-[3-[tert-butoxycarbonyl(methyl)amino]prop-l-ynyl]-2-fluorophenoxy]propyl]-2-[[5-[tert-butyl (dimethyl)silyl]oxy-4-methoxy-pentyl]-[5-methyl-6-[(Z)[3-(2-trimethylsilylethoxymethyl)-l,3-benzothiazol-2-ylidene]amino]pyridazin-35 yl]amino]thiazole-4-carboxylate
Using Buchwald General Procedure II starting from 12 g of Préparation 3f (13 mmol) and 6.30 g of Préparation 4a (15.6 mmol) as the appropriate halide, 14 g (83%) of the desired product was obtained.
Step B: 2-[[6-(l,3-benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-(5-hydroxy-4-methoxy10 pentyl)amino]^5-[3-[2-fluoro-4-[3-(methylamino)prop-l-ynyl]phenoxy]propyl]thiazole-4carboxylic acid
Using Deprotection and Hydrolysis General Procedure starting from the product from Step A as the appropriate methyl ester, the desired product was obtained.
'H NMR (500 MHz, DMSO-d6) δ ppm 7.91 (d, 1H), 7.66 (s, 1H), 7.53 (d, 1H), 7.39 (td, 1H), 15 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); 13C NMR (125 MHz, DMSO-d6) 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, 17.8 HRMS-ESI (m/z): [M+H]+ calcd for C35H39FN7O5S2: 720.2433, found 20 720.2437.
Example 113:2-[[6-(l,3-Benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-(4-hydroxy-5morpholino-pentyl)amino]-5-[3-[2-fluoro-4-[3-(methylamino)prop-lynyl]phenoxy]propyl] thiazole-4-carboxylic acid
363
Step A: methyl5-[3-[4-[3-[tert-butoxycarbonyl(methyl)amino]prop-l-ynyl]-2-fluorophenoxy]propyl]-2-[[4-[tert-butyl(diphenyl)silyl]oxy-5-morpholino-pentyl]-[5-methyl-6-[(Z)[3-(2-trimethylsilylethoxymethyl)-l,3-benzothiazol-2-ylidene]amino]pyridazin-35 yl]amino]thiazole-4-carboxylate
Using Alkylation with tosylate General Procedure starting from Préparation 5a and morpholine as the appropriate amine, the desired product was obtained.
Step B: 2-[[6-(1,3-benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-(4-hydroxy-5morpholino-pentyl)amino]-5-[3-[2-fluoro-4-[3-(methylamino)prop-l-ynyl]phenoxy]propyl] 10 thiazole-4-carboxylic acid
Using Deprotection and Hydrolysis General Procedure starting from the product from Step A as the appropriate methyl ester, the desired product was obtained.
HRMS-ESI (m/z): [M+H]+ calcd for C38H44FN8O5S2: 775.2855, found 775.2851.
Example 114: 2-({6-[(l,3-Benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(415 hydroxybutyI)amino)-5-(3-{2-fluoro-4-[3-(methylamino)prop-l-yn-lyl]phenoxy}propyl)-l,3-thiazole-4-carboxylic acid
364
Using Deprotection and Hydrolysis General Procedure starting from the appropriate methyl ester Préparation 5c, Step A, the desired product was obtained.
HRMS-ESI (m/z): [M+H]+ calcd for C33H35FN7O4S2: 676.2170, found 676.2177.
Example ll5^2-rr6-(l.3-Benzothiazol-2-vlamino)-5-methvl-pvridazin-3-vl1-(3-hydroxv-25 morpholino-propyl)amino]-5-[3-[2-fluoro-4-[3-(methylamino)prop-lyny 1] phenoxy] propyl] thiazoIe-4-carboxylic acid
Step A: methyl5-[3-[4-[3-[tert-butoxycarbonyl(methyl)amino]prop-l-ynyl]-2-fluorophenoxyjpropyl]-2-[/3-1tert-butyl(dimethyl)silyl]oxy-2-morpholino-propyl]-[5-methyl-610 [(Z)-[3-(2-trimethylsilylethoxymethyl)-l,3-benzothiazol-2-ylidene]amino]pyridazin-3yl]amino]thiazole-4-carboxylate
Using Buchwald General Procedure II starting from 390 mg of Préparation 3r (0.59 mmol, 1 eq.) and 216 mg of Préparation 4a (0.59 mmol, 1 eq.) as the appropriate halide, 500 mg (83%) of the desired product was obtained.
XH NMR (500 MHz, DMSO-dô) 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); 13C NMR (125 MHz, DMSO-dô) 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; HRMS-ESI (m/z): [M+H]+ calcd for C54H78FN8O8S2Si2: 1105.4901, found 1105.4896.
Step B: 2-[[6-(l,3-benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-(3-hydroxy-2I
365 morpholino-propyl)amino]-5-[3-[2-fluoro-4-[3-(methylamino)prop-lynyl]phenoxy]propyl]thiazole-4-carboxylic acid
Using Deprotection and Hydrolysis General Procedure starting from the product from Step A as the appropriate methyl ester, the desired product was obtained.
HRMS-ESI (m/z): [M+H]+ calcd for C36H40FN8O5S2: 747.2542, found 747.2531.
Example 116: 2-({6-[(l,3-Benzothiazol-2-yl)amino]-5-methylpyridazin-3yl}(methyl)amino)-5-(3-{4-[3-(dimethylamino)prop-l-yn-l-yl]-2-fluorophenoxy}-2methylpropyl)-l,3-thiazole-4-carboxy!ic acid
Step A: ethyl 2-[methyl(5-methyl-6-{[(2Z)-3-{[2-(trimethylsilyl)ethoxy]methyl}-2,3-dihydrol,3-benzothiazol-2-ylidene]amino}pyridazin-3-yl)amino]-5-(2-methyl-3-oxopropyl)-l,3thiazole-4-carboxylate
To a solution of the product from Préparation 11b (1.2 g, 1.89 mmol, 1 eq) in 1,4-dioxane (20 mL) was added 2-methyl-2-propen-l-ol (0.48 mL, 5.66 mmol, 3 eq), N,Ndicyclohexylmethylamine (0.81 mL, 3.78 mmol, 2 eq) and 2-(di-ierLbutyl-phosphino)-lphenyl-l/7-pyrrole (54.3 mg, 0.19 mmol, 0.1 eq). The mixture was sparged 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 sealed flask overnight. The reaction was partitioned between ethyl acetate and water, and the organic phase was washed with brine, dried (magnésium sulfate), and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 40 g RediSep™ silica cartridge) eluting with a gradient of 0 - 40% ethyl acetate in Ao-heptane afforded the desired product as a yellow oil (779 mg, 1.24 mmol, 66%).
366
LC/MS (C29H38N6O4S1S2) 627 [M+H]+; RT 2.86 (LCMS-V-C)
Ή NMR (400 MHz, DMSO-d6) 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-{[(2Z)-3-{[2(trimethylsilyi)ethoxy]methyl}-2,3-dihydro-l,3-benzothiazol-2-ylidene]amino}pyridazin-3yl)amino]-l,3-^hiazole-4-carboxylate
To a solution of the product of Step A (779 mg, 1.24 mmol, 1 eq) in methanol (20 mL), cooled to 0 °C was added sodium borohydride (51.7 mg, 1.37 mmol, 1.1 eq) and the mixture was stirred for 20 min. The reaction was partitioned between ethyl acetate and water, and the organic phase was washed with brine, dried (magnésium sulfate), and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash RT, 24 g RediSep™ silica cartridge) eluting with a gradient of 0 - 50% ethyl acetate in Ao-heptane afforded the desired product as a yellow foam (336 mg, 0.53 mmol, 43%).
LC/MS (C29H4oN604SiS2) 629 [M+H]+; RT 2.74 (LCMS-V-C) I >H NMR (400 MHz, DMSO-d6) 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), 31.75 - 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-{[(2Z)-3-{[2(trimethylsilyl}ethoxy]methyl}-2,3-dihydro-l,3-benzothiazol-2-ylidene]amino}pyridazin-3yl)amino]-l,3-thiazole-4-carboxylate
To a solution of the product of Step B (469 mg, 0.75 mmol, 1 eq) in 6:1 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 température for 16 h. The reaction was partitioned between ethyl acetate and 10% aqueous sodium thiosulphate, and the organic phase was dried (magnésium sulfate) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 12 g RediSep™ silica cartridge) eluting with a gradient of 0 - 30% ethyl acetate in isoheptane afforded the desired product as a yellow oil (523 mg, 0.71 mmol, 95%).
367
LC/MS (C29H39IN6O3S1S2) 739 [M+H]+; RT 3.23 (LCMS-V-C)
Ή NMR (400 MHz, DMSO-d6) 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-l-yn-1 -yl]-2-fluorophenoxy}-2methylpropyl)-2-[methyl(5-methyl-6-{[(2Z)-3-{[2-(trimethylsilyl)ethoxy]methyl}-2,3dihydro-l,3-benzothiazol-2-ylidene]amino}pyridazin-3-yl)amino]-l,3-thiazole-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 from Préparation 6b (178 mg, 0.92 mmol, 1.3 eq) and césium carbonate (692 mg, 2.12 mmol, 3 eq) and the mixture was heated at 80 °C for 3 h. The reaction was partitioned between ethyl acetate and brine, and the organic phase was dried (magnésium sulfate) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 24 g RediSep™ silica cartridge) eluting with a gradient of 0 - 5% methanol in dichloromethane afforded the desired product as a yellow oil (257 mg, 0.32 mmol, 45%).
LC/MS (C4oH5oFN704SiS2) 804 [M+H]+; RT 2.66 (LCMS-V-C)
Ή NMR (40Q MHz, DMSO-d6) 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-[(l,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)5-(3-{4-[3-(dimethylamino)prop-l-yn-l-yl]-2-fluorophenoxy}-2-methylpropyl)-l,3-thiazole4-carboxylate
To a solution of the product of Step D (257 mg, 0.32 mmol, 1 eq) in tetrahydrofuran (15 mL) was added ethylenediamine (64 pL, 0.96 mmol, 3 eq) followed by TBAF (IM in tetrahydrofurah; 0.96 mL, 0.96 mmol, 3 eq) and the mixture was heated at 60 °C for 18 h. The reaction was partitioned between ethyl acetate and water, and the organic phase was dried (magnésium sulfate), and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 12 g RediSep™ silica cartridge) eluting with a gradient of 0
368
- 5% methanol in dichloromethane afforded the desired product as a yellow glass (110 mg, 0.16 mmol, 51%).
LC/MS (C34H36FN7O3S2) 674 [M+H]+; RT 2.11 (LCMS-V-C) 'H NMR (400 MHz, DMSO-d6) δ 11.55 (br s, 1H), 7.92 (d, J = 8.0 Hz, 1H), 7.69 (s, 1H), 7.54 5 (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), 2.40 - 2.27 (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-[(l,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)-5-(3- {4-[3-(dimethylamino)prop-l -yn-1 -yl]-2-fluorophenoxy}-2-methylpropyl)-l,3-thiazole-4- carboxylic acid
To a solution of the product of 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 heated at reflux for 16 h. The reaction was concentrated in vacuo and the residue was triturated with 15 water (5 mL), filtered, washed with water and dried under vacuum to afford the desired product as a yellow solid (91.2 mg, 0.14 mmol, 87%) [as a lithium sait].
HRMS-ESI (m/z) [M+H]+ calcd for C32H33FN7O3S2: 646.2065, found 646.2091.
Example 117:2-[[6-(l,3-Benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-(2,3dihydroxypropyl)amino]-5-[3-[2-fluoro-4-[3-(methylamino)prop-l20 ynyl]phenoxy] propyl] thiazole-4-carboxylic acid
Step A: methyl5-[3-[4-[3-[tert-butoxycarbonyl(methyl)amino]prop-l-ynyl]-2-fluorophenoxy]propyl]-2-[(2,2-dimethyl-l,3-dioxolan-4-yl)methyl-[5-methyl-6-[(Z)-[3-(2trimethylsilylethoxymethyl)-l,3-benzothiazol-2-ylidene]amino]pyridazin-325 yl]amino]thiazole-4-carboxylate
369
Using Buchwald General Procedure II starting from 280 mg of Préparation 3o (0.47 mmol, 1 eq.) and 193 mg of Préparation 4a (0.47 mmol, 1 eq.) as the appropriate halide, 400 mg (88%) of the desired product was obtained.
XH NMR (500 MHz, DMSO-d6) 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); 13C NMR(125MHz, DMSO-d6)ôppm 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; HRMS-ESI (m/z): [M+H]+ calcd for C47H6iFN7O8S2Si: 962.3771, found 962.3767.
Step B: 2-[[6-(l,3-benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-(2,3dihydroxypro^yl)amino]-5-[3-[2-fluoro-4-[3-(methylamino)prop-l ynyl]phenoxy]propyl]thiazole-4-carboxylic acid
Using Deprotection and Hydrolysis General Procedure starting from the product from Step A as the appropriate methyl ester, the desired product was obtained.
HRMS-ESI (m/z): [M+H]+ calcd for C32H33FN7O5S2: 678.1963, found 678.1965.
Example 118:2-[[6-(l,3-Benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yI]-(3-hydroxy-2methoxy-propyl)amino]-5- [3- [2-fluoro-4- [3-(methylamino)prop-l-ynyl] phenoxy]propyl] thiazole-4-carboxylic acid
Step A: methyl 5-[3-[4-[3-[tert-butoxycarbonyl(methyl)amino]prop-l-ynyl]-2-fluorophenoxy]propyl]-2-[[3-[tert-butyl(dimethyl)silyl]oxy-2-methoxy-propyl]-[5-methyl-6-[(Z)-[3(2-trimethylsilylethoxymethyl)-l,3-benzothiazol-2-ylidene]amino]pyridazin-320487
370 yl]amino]thiazole-4-carboxylate
Using Buchwald General Procedure II starting from 400 mg of Préparation 3q (0.59 mmol, 1 eq.) and 240 mg of Préparation 4a (0.59 mmol, 1 eq.) as the appropriate halide, 534 mg (86%) of the desired product was obtained.
Ή NMR (500 MHz, DMSO-d6) 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); 13C NMR (125 MHz,
DMSO-d6)ôppm 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, 28.4, 26.2, 23.0,18.0,17.7, -1.6, -5.1, -5.1; HRMS-ESI (m/z): [M+H]+ calcd for C51H73FN7O8S2S12: 1050.4479, found 1050.4469.
Step B: 2-[[6-(l,3-benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-(3-hydroxy-2-methoxypropyl)amino]-5-[3-[2-fluoro-4-[3-(methylamino)prop-l-ynyl]phenoxy]propyl] thiazole-415 carboxylic acid
Using Deprotection and Hydrolysis General Procedure starting from the product from Step A as the appropriate methyl ester, the desired product was obtained.
HRMS-ESI (m/z): [M+H]+ calcd for C33H35FN7O5S2: 692.2120, found 692.2117.
Example 119:2-[[6-(l,3-Benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-(4,520 dihydroxypentyl) amino]-5-[3-[2-fluoro-4-[3-(methylamino)prop-lyny!]phenoxy]propyl]thiazole-4-carboxylic acid
371
Using Deprotection and Hydrolysis General Procedure starting from the product from Préparation 5a, Step A as the appropriate methyl ester, the desired product was obtained.
HRMS-ESI (m/z): [M+H]+ calcd for C34H37FN7O5S2: 706.2276, found 706.2274.
Example 120:2-[[6-(l,3-Benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-(3,45 dihydroxybutyl)amino]-5-[3-[2-fluoro-4-[3-[methyl(p-tolylsulfonyl)amino]prop-lyny 1] phenoxy] propyl] thiazole-4-carboxylic acid
Step A: methyl2-[2-(2,2-dimethyl-l,3-dioxolan-4-yl)ethyl-[5-methyl-6-[(Z)-[3-(2trimethylsilylethoxymethyl)-l,3-benzothiazol-2-ylidene]amino]pyridazin-3-yl]amino]-5-[310 [2-fluoro-4-[3-[methyl(p-tolylsulfonyl)amino]prop-l-ynyl]phenoxy]propyl]thiazole-4carboxylate
Using Buchwald General Procedure II starting from 470 mg of Préparation 3k (0.71 mmol, 1 eq.) and 290 mg of Préparation 4a (0.71 mmol, 1 eq.) as the appropriate halide, 660 mg (90%) of the desired product was obtained.
Step B: 2-[[6-(l,3-benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-(3,4dihydroxybutyl)amino]-5-[3-[2-fluoro-4-[3-[methyl(p-tolylsulfonyl)amino]prop-lynyl]phenoxy]propyl]thiazole-4-carboxylic acid
Using Deprotection and Hydrolysis General Procedure starting from the product from Step A as the appropriate methyl ester, the desired product was obtained.
HRMS-ESI (m/z): [M+H]+ calcd for C40H41FN7O7S3: 846.2208, found 846.2201.
372
Example 121:2-[[6-(l,3-Benzothiazol-2-ylamino)-5-methyI-pyridazin-3-yl]-(3,4dihydroxybutyl)ammo]-5-[3-[2-fluoro-4-[3-[(4-methoxyphenyl)methyl-methylamino]prop-l-ynyl]phenoxy]propyl]thiazole-4-carboxylic acid
Step A: methyl 2-[2-(2,2-dimethyl-l,3-dioxolan-4-yl)ethyl-[5-methyl-6-[(Z)-[3-(2trimethylsilylethoxymethyl)-l,3-benzothiazol-2-ylidene]amino]pyridazin-3-yl]amino]-5-[3[2-fluoro-4-[3-[(4-methoxyphenyl)methyl-methyl-amino]prop-lynyl]phenoxy]propyl]thiazole-4-carboxylate
Using Buchwald General Procedure II starting from 625 mg of Préparation 3j (1 mmol, 1 eq.) and 407 mg of Préparation 4a (1 mmol, 1 eq.) as the appropriate halide, 550 mg (55%) of the desired product was obtained.
Step B: 2-[[6-(l,3-benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-(3,4dihydroxybutyl)amino]-5-[3-[2-fluoro-4-[3-[(4-methoxyphenyl)methyl-methyl-amino]prop-
-ynyl]phenoxy]propyl]thiazole-4-carboxylic acid
Using Deprotection and Hydrolysis General Procedure starting from the product from Step A as the appropriate methyl ester, the desired product was obtained.
HRMS-ESI (m/z): [M+H]+ calcd for C41H43FN7O6S2: 812.2695, found 812.2674.
Example 122:2-[[6-(l,3-Benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-(2,3dihydroxypropyl)amino]-5-[3-[4-[3-(dimethylamino)prop-l-ynyl]-2-fluorophenoxy] propyl] thiazole-4-carboxy lie acid
Step A: methyl5-[3-[4-[3-(dimethylamino)prop-l-ynyl]-2-fluoro-phenoxy]propyl]-2-[(2,2dimethyl-l,3-dioxolan-4-yl)methyl-[5-methyl-6-[(Z)-[3-(2-trimethylsilylethoxymethyl)-l,3benzothiazol-2-ylidene]amino]pyridazin-3-yl]amino]thiazole-4-carboxylate
Using Buchwald General Procedure II starting from 380 mg of Préparation 3p (0.75 mmol, 1 eq.) and 306 mg of Préparation 4a (0.75 mmol, 1 eq.) as the appropriate halide, 480 mg (72%) of the desired product was obtained.
Ή NMR (500 MHz, DMSO-d6) 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), 10 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); 13C NMR (125 MHz, DMSO-d6) 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, 72.7, 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; HRMS-ESI (m/z): [M+H]+ calcd 15 for C43H55FN7O6S2Si: 876.3403, found 876.3408.
Step B: 2-[[6-(l,3-benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-(2,3dihydroxypropyl)amino]-5-[3-[4-[3-(dimethylamino)prop-l-ynyl]-2-fluorophenoxy]propyl]thiazole-4-carboxylic acid
Using Deprotection and Hydrolysis General Procedure starting from the product from Step 20 A as the appropriate methyl ester, the desired product was obtained.
HRMS-ESI (m/z): [M+H]+ calcd for C33H35FN7O5S2: 692.2120, found 692.2117.
Example 123:2-ir6-(1.3-Benzothiazol-2-ylamino)-5-methyI-pyridazin-3-yl]-(3,4dihydroxybutyl)amino]-5-[3-[4-[3-(dimethylamino)prop-l-ynyl]-2-fluorophenoxy]propyl]thiazole-4-carboxylic acid
374
Step A: methyl5-[3-[4-[3-(dimethylamino)prop-l-ynyl]-2-fluoro-phenoxy]propyl]-2-[2-(2,2dimethyl-l,3-dioxolan-4-yl)ethyl-[5-methyl-6-[(Z)-[3-(2-trimethylsilylethoxymethyl)-l,3benzothiazol-2-ylidene]amino]pyridazin-3-yl]amino]thiazole-4-carboxylate
Using Buchwald General Procedure II starting from 350 mg of Préparation 3i (0.67 mmol, 1 eq.) and 275 mg of Préparation 4a (0.67 mmol, 1 eq.) as the appropriate halide, 510 mg (85%) of the desired product was obtained.
Step B: 2-[[6-(l,3-benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-(3,4dihydroxybutyl)amino]-5-[3-[4-[3-(dimethylamino)prop-l-ynyl]-2-fluoro10 phenoxy]propyl]thiazole-4-carboxylic acid
Using Deprotection and Hydrolysis General Procedure starting from the product from Step A as the appropriate methyl ester, the desired product was obtained.
HRMS-ESI (m/z): [M+H]+ calcd for C34H37FN7O5S2: 706.2276, found 706.2270.
Example 124:2-[[6-(l,3-Benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yI]-[5-[215 (dimethylamino)ethyl-methyl-amino]-4-hydroxy-pentyl]amino]-5-[3-[4-[3(dimethylamino)prop-l-ynyl]-2-fluoro-phenoxy]propyl]thiazole-4-carboxylic acid
375
Step A: methyl 2-[[4-[tert-butyl(diphenyl)silyl]oxy-5-[2-(dimethylamino)ethyl-methylamino]pentyl]-[5-methyl-6-[(Z)-[3-(2-trimethylsilylethoxymethyl)-l,3-benzothiazol-2ylidene]amino]pyridazin-3-yl]amino]-5-[3-[4-[3-(dimethylamino)prop-l-ynyl]-2-fluoro5 phenoxy]propyl]thiazole-4-carboxylate
Using Alkylation with tosylate General Procedure starting from Préparation 5b and N,N’,N'-trimethylethane-l,2-diamine as the appropriate amine, the desired product was obtained.
Step B: 2-[[6-(l,3-benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-[5-[210 (dimethylamino) ethyl-methyl-amino]-4-hydroxy-pentyl]amino]-5-[3-[4-[3(dimethylamino)prop-l-ynyl]-2-fluoro-phenoxy]propyl]thiazole-4-carboxylic acid
Using Deprotection and Hydrolysis General Procedure starting from the product from Step A as the appropriate methyl ester, the desired product was obtained.
HRMS-ESI (m/z): [M+H]+ calcd for C40H51FN9O4S2: 804.3484, found 804.3487.
Example 125:2-Fr6-(l,3-Benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-(5hydroxypentyl)amino]thiazole-4-carboxylic acid
376
Step A: methyl 2-[tert-butoxycarbonyl-[5-[tertbutyl(dimethyl)silyl]oxypentyl]amino]thiazole-4-carboxylate
Using Mitsunobu General Procedure starting from methyl 2-(tert5 butoxycarbonylamino)thiazole-4-carboxylate and 5-[tert-butyl(dimethyl)silyl]oxypentan-l -ol as starting materials in THF at rt for 2 h, 565 mg (82%) of the desired product was obtained.
*H NMR (400 MHz, DMSO-d6) 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 Deprotection with HFIP General Procedure starting from the product from Step A as the appropriate carbamate, 484 mg (113%) of the desired product was obtained.
*H NMR (400 MHz, DMSO-d6) 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-[(Z) -[3-(215 trimethylsilylethoxymethyl)-l,3-benzothiazol-2-ylidene]amino]pyridazin-3yl]amino]thiazole-4-carboxylate
Using Buchwald General Procedure II starting from the product from Step B and Préparation 4a as the appropriate halide, 535 mg (61%) of the desired product was obtained.
Ή NMR (400 MHz, DMSO-ife) δ ppm 7.97 (s, 1H), 7.84 (d, 1H), 7.70 (s, 1H), 7.46 (t, 1H), 20 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).
377 1 Step D: 2-[[6-(l,3-benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-(5hydroxypentyl)amino]thiazole-4-carboxylic acid
Using Deprotection and Hydrolysis General Procedure followed by repurification via reverse phase préparative chromatography (C18, 25 mM NH4HCO3 water : MeCN) starting 5 from the product from Step C, the desired product was obtained.
HRMS-ESI (m/z): [M+H]+ calcd for C21H23N6O3S2: 471.1273, found: 471.1271.
Example 126:2-[[6-(l,3-Benzothiazol-2-ylamino)-5-methyI-pyridazin-3-yl]-(3hydroxypropyl)amino]-5-[3-[4-[3-(dimethylammo)prop-l-ynyl]-2-fluorophenoxy]propyl]thiazole-4-carboxylic acid
Step A: ethyl3-bromo-6-chloro-2-oxo-hexanoate
To 2 g of ethyl 6-chloro-2-oxo-hexanoate (10.4 mmol, 1 eq.) in 20 mL of CCI4 was added 0.6 mL of bromine (11.5 mmol, 1.1 eq.) dropwise and the mixture was stirred at rt for 1 h. After pouring the mixture into 50 mL of a 1 M aqueous solution Na2S2O3, it was extracted with 2x50 15 mL of 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 give 2.05 g (73%) of the desired product.
Tl NMR (400 MHz, DMSO-d6) 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/LR-EI: [M]+ = 270.
Step B: 3-[tert-butyl(dimethyl)silyl]oxypropan-l-amine
To 8.25 g of 3-aminopropan-l-ol (110 mmol, 1 eq.), 23 mL of N,N-diethylethanamine (170 mmol, 1.5 eq.), and 30 mg of imidazole (0.44 mmol, 0.004 eq.) in 200 mL of DCM was added
378
18.2 g of tert-butyldimethylsilyl chloride (150 mmol, 1.4 eq.) and the mixture was stirred at rt for 15 h. After pouring the reaction mixture onto 250 mL of water and extracting with 2x200 mL of DCM, the combined organic layers were washed with 50 mL of water and 50 mL of brine, dried, and concentrated to give 20.9 g (100%) of the desired product.
JH NMR (400 MHz, DMSO-î/6) 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 was added dropwise a solution of 3.5 g of the product from Step B (18.5 mmol, 1 eq.) in 10 mL of DCM and the mixture was stirred at rt for 2 h. After concentration, the crude product was purified by silica gel chromatography using Petroleum ether and EtOAc as eluents to give 3.87 g (80%) of the desired product.
Ή NMR (400 MHz, DMSO-î/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 H); 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 éthanol was heated to reflux for 18 h. 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 give 10.1 g (81%) of the desired product.
Ή NMR (400 MHz, DMSO-î/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-4methylpyridazine (35.6 mmol, 1.5 eq.) in 120 mL of THF was added 2.1 g of sodium hydride (52.1 mmol, 2.2 eq.) portion wise at 50 °C. The chilled mixture was poured into ice and extracted with 2x200 mL of ethyl acetate. The combined organic layers were washed with brine, dried, concentrated, and purified by silica gel chromatography using petroleum ether and
379
EtOAc as eluents to give 4.3 g (32%) of the desired product.
Ή NMR (400 MHz, DMSO-d6) 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-methyl-pyridazin-3yl)amino]-5-(3-iodopropyl)thiazole-4-carboxylate
The 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 h. After concentration, the residue was taken up in ethyl acetate, washed with water and brine, dried, and concentrated to give the desired product, which was used without further purification in the next step.
XH NMR (400 MHz, DMSO-î/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-methyl-pyridazin-3yl)amino]-5-[3-[4-[3-(dimethylamino)prop-l-ynyl]-2-fluoro-phenoxy]propyl]thiazole-4carboxylate
The mixture of 800 mg of the product from Step F (1.25 mmol, 1 eq.), 821 mg of césium carbonate (2.5 mmol, 2 eq.) and 290 mg of Préparation 6b (1.5 mmol, 1.2 eq.) in 6 mL of MeCN was heated to reflux for 2 h. After dilution with 100 mL of ethyl acetate, the organic layer was washed with 50 mL of water and 50 mL of brine, dried, concentrated, and was purified by silica gel chromatography using DCM and MeOH (containing 7 N NH3) as eluents to give 500 mg (57%) of the desired product.
Ή NMR (400 MHz, DMSO-tZe) 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-(l,3-benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-(3hydroxypropyl)amino]-5-[3-[4-[3-(dimethylamino)prop-l-ynyl]-2-fluorophenoxy]propyl]thiazole-4-carboxylate
The mixture of 100 mg of the product from Step G (0.142 mmol, 1 eq.), 28 mg of 1,3benzothiazol-2-amine (0.185 mmol, 1.3 eq.), 10 mg of 4,5-bis(diphenylphosphino)-9,920487
380 dimethylxanthene (0.017 mmol, 0.12 eq.), 8 mg of tris(dibenzylideneacetone)dîpalladium(0) (8.52 pmol, 0.06 eq.), and 72 pL of DIPEA (0.426 mmol, 3 eq.) in 0.7 mL of N-methyl-2pyrrolidone was kept at 120 °C for 18 h. After pouring the mixture into 50 mL of DCM, the organic layer was washed with 10 mL of water and 10 mL of a saturated aqueous solution of 5 lithium chloride, dried, concentrated, and purified by silica gel chromatography using DCM and MeOH (containing 7 N NH3) as eluents to afford 40 mg (40%) of the desired product.
Hl NMR (400 MHz, DMSO-d6) 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 10 (m, 2H), 1.87 (m, 2H), 1.29 (t, 3H); IR: 3414, 3400-2200, 1709, 1602,1149/1087.
Step I: 2-[[6-(l,3-benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-(3hydroxypropyl)amino]-5-[3-[4-[3-(dimethylamino)prop-l-ynyl]-2-fluorophenoxy]propyl]thiazole-4-carboxylic acid
The mixture of the product from Step H and LiOHxH2O (4 eq.) in 1,4-dioxane and water was 15 stirred at rt for 3 h. After the addition of an extra 1 eq. of LiOHxH2O in water, the reaction was stirred at rt for 18 h. After the slow addition of 2 N solution of HCl, the reaction mixture was concentrated and purified via reverse phase préparative chromatography (0.2 M NH4CO3 in water : MeCN) to give the desired product.
HRMS-ESI (m/z): [M+H]+ calcd for C33H35FN7O4S2: 676.2170 found: 676.2169.
Example 127:2-IT6-(L3-Benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-(3hydroxypropyl)amino]-5-[3-[2-fluoro-4-[3-(methylamino)prop-lynyl]phenoxy]propyl]thiazole-4-carboxylic acid
Step A: methyl5-[3-[4-[3-[tert-butoxycarbonyl(methyl)amino]prop-l-ynyl]-2-fluoro
381 phenoxy]propyl]-2-[3-[tert-butyl(dimethyl)silyl]oxypropyl-[5-methyl-6-[(Z)-[3-(2trimethylsilylethoxymethyl)-l,3-benzothiazol-2-ylidene]amino]pyridazin-3yl]amino]thiazole-4-carboxylate
Using Buchwald General Procedure II starting from 300 mg of Préparation 3n (0.46 mmol, 5 1 eq.) and 187 mg of Préparation 4a (0.46 mmol, 1 eq.) as the appropriate halide, 395 mg (83%) of the desired product was obtained.
‘H NMR (500 MHz, DMSO-d6) 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); 13C
NMR (125 MHz, DMSO-d6) 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; HRMS-ESI (m/z): [M+H]+ calcd for C50H71FN7O7S2S12: 1020.4373, found 1020.4365.
Step B: 2-[[6-(l,3-benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-(3hydroxypropyl)amino]-5-[3-[2-fluoro-4-[3-(methylamino)prop-lynyl]phenoxy]propyl]thiazole-4-carboxylic acid
Using Deprotection and Hydrolysis General Procedure starting from the product from Step A as the appropriate methyl ester, the desired product was obtained.
HRMS-ESI (m/z): [M+H]+ calcd for C32H33FN7O4S2: 662.2014, found 662.2016.
Example 128:2-[[6-(l,3-Benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-(5hydroxypentyl)amino]-5-[3-[2-fluoro-4-[3-(methylamino)prop-lynyl]phenoxy]propyl]thiazole-4-carboxylic acid
382
Using Deprotection and Hydrolysis General Procedure starting from the appropriate methyl ester Préparation 5e, Step A, the desired product was obtained.
HRMS-ESI (m/z): [M+H]+ calcd for C34H37FN7O4S2: 690.2327, found 690.2318.
Example 130:3-[3-[4-[3-[2-[[6-(l,3-BenzothiazoI-2-ylamino)-5-methyl-pyridazin-3-yl]-(45 hydroxybutyl)amino]-4-carboxy-thiazol-5-yl]propoxy]-3-fluoro-phenyi]prop-2-ynyldimethyl-ammonio]propane-l-sulfonate
Step A: 3-[3-[4-[3-[2-[4-[tert-butyl(dimethyl)silyl]oxybutyl-[5-methyl-6-[(Z)-[3-(2trimethylsilyl- ethoxymethyl)-l,3-benzothiazol-2-ylidene]amino]pyridazin-3-yl]amino]-410 methoxycarbonyl-thiazol-5-yl]propoxy]-3-fluoro-phenyl]prop-2-ynyl-dimethylammonio]propane-l -sulfonate
To a suspension of 660 mg of Préparation 5d, Step A (0.7 mmol, 1 eq.) in 5 mL 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 rt. The crude product was purified by flash column 15 chromatography to obtain 550 mg (75%) of the desired product.
Ή NMR (500 MHz, DMSO-d6) 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);
13C NMR (125 MHz, DMSO-d6) 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-ESI (m/z): [M+2H]2+ calcd for C5oH74FN708S3Si2: 535.7136, found 535.7136.
Step B: 3-[3-[4-[3-[2-[[6-(l,3-benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-(420487
383 hydroxybutyl)amino]-4-carboxy-thiazol-5-yl]propoxy]-3-fluoro-phenyl]prop-2-ynyldimethyl-ammonio]propane-l-sulfonate
Using Deprotection and Hydrolysis General Procedure starting from the product from Step A as the appropriate methyl ester, the desired product was obtained.
HRMS-ESI (m/z): [M+H]+ calcd for C37H43FN7O7S3: 812.2365, found 812.2357.
Example 131:2-[[6-(l,3-Benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yI]-(4-hydroxy-5morpholino-pentyl)amino]thiazole-4-carboxylic acid
Step A: methyl 2-[tert-butoxycarbonyl-[4-[tert-butyl(dimethyl)silyl]oxy-5-morpholinopentyl]amino]thiazole-4-carboxylate
Using Mitsunobu General Procedure starting from 639 mg of methyl 2-(tertbutoxycarbonylamino)thiazole-4-carboxylate (2.47 mmol, 1.5 eq.) and 500 mg of Préparation 2e (1.65 mmol, 1 eq.) as the appropriate alcohol, 730 mg (81%) of the desired product was obtained.
!H NMR (400 MHz, DMSO-d6) δ ppm 8.08 (s, 1H), 4.09-4.03 (m, 2H), 3.79 (s+m, 4H), 3.51 (t, 4H), 2.39-2.28 (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); 13C NMR (100 MHz, DMSOd6) 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]+ calcd for C25H46N3O6SSi: 544.3, found 544.3.
Step B: methyl 2-[[4-[tert-butyl(dimethyl)silyl]oxy-5-morpholino-pentyl]amino]thiazole-4carboxylate
384
Using Deprotection with HFIP General 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.
Ή NMR (400 MHz, DMSO-rfc) 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); 13C NMR (100 MHz, DMSO-rfc) 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]+ calcd for C20H38N3O4SSi: 444.2, found 444.3.
Step C: methyl 2-[[4-[tert-butyl(dimethyl)silyl]oxy-5-morpholino-pentyl]-[5-methyl-6-[(Z)[3-(2-trimethylsilylethoxymethyl)-l,3-benzothiazol-2-ylidene]amino]pyridazin-3yl]amino]thiazole-4-carboxylate
Using Buchwald General Procedure II starting from 231 mg of the product from Step B (0.52 mmol, 1 eq.) and 315 mg of Préparation 4a (0.77 mmol, 1.5 eq.) as the appropriate halide, 344 mg (81%) of the desired product was obtained.
Ή NMR (500 MHz, DMSO-ifc) 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), 1.56-1.41 (m+m, 2H), 0.91 (t, 2H), 0.75 (s, 9H), 0.01-(-0.05) (s+s, 6H), -0.12 (s, 9H); 13C NMR (125 MHz, DMSO-c/e) 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; HRMS-ESI (m/z): [M+H]+ calcd for C38H6oN705S2Si2: 814.3630, found 814.3629.
Step D: 2-[[6-(l,3-benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-(4-hydroxy-5morpholino-pentyl)amino]thiazole-4-carboxylic acid
Using Deprotection and Hydrolysis General Procedure followed by purification via reverse phase préparative chromatography (C18, 25 mM NH4HCO3 in water : MeCN) starting from the product from Step C, the desired product was obtained.
HRMS-ESI (m/z): [M+H]+ calcd for C25H3oN704S2: 556.1795, found 556.1807.
Example 132:2-[[6-(l,3-Benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-(4-hydroxy-5methoxy-pentyl)amino]thiazole-4-carboxylic acid
385
Step A: methyl 2-[tert-butoxycarbonyl-(5-methoxy-4-triisopropylsilyloxypentyl)amino]thiazole-4-carboxylate
Using Mitsunobu General Procedure starting from 284 mg of methyl 2-(tert5 butoxycarbonylamino)thiazole-4-carboxylate (1.10 mmol, 1.1 eq.) and 291 mg of Préparation 2c (1.00 mmol, 1 eq.) as the appropriate alcohol, 456 mg (86%) of the desired product was obtained.
Ή NMR (400 MHz, DMSO-de) 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); 13C 10 NMR (100 MHz, DMSO-d6) 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]+ calcd for C25H47N2O6SSi: 531.3, found 531.4.
Step B: methyl 2-[(5-methoxy-4-triisopropylsilyloxy-pentyl)amino]thiazole-4-carboxylate
Using Deprotection with HFIP General Procedure starting from 444 mg of the product from 15 Step A (0.84 mmol) as the appropriate carbamate, 334 mg (93%) of the desired product was obtained.
XH NMR (500 MHz, DMSO-î/6) 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); 13C NMR (125 MHz, DMSO-*) δ ppm 162.0, 116.9, 76.7, 71.2, 58.9, 52.0, 45.0, 32.2, 24.4; HRMS-ESI 20 (m/z): [M+H]+ calcd for C2oH39N204SSi: 431.2394, found 431.2392.
Step C: methyl 2-[(5-methoxy-4-triisopropylsilyloxy-pentyl)-[5-methyl-6-[(Z)-[3-(2trimethylsilylethoxymethyl)-l,3-benzothiazol-2-ylidene]amino]pyridazin-3yl]amino]thiazole-4-carboxylate
Using Buchwald General Procedure II starting from 324 mg of the product from Step B (0.75 25 mmol, 1 eq.) and 337 mg of Préparation 4a (0.83 mmol, 1.1 eq.) as the appropriate halide, 540
386 mg (90%) of the desired product was obtained.
Ή NMR (400 MHz, DMSO-dô) 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); 13C NMR (125 MHz, 5 DMSO-d6) 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 HRMS-ESI (m/z): [M+H]+ calcd for C38H6iN6O5S2Si2: 801.3678, found 801.3671.
Step D: 2-[[6-(l,3-benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-(4-hydroxy-5-methoxypentyl)amino]thiazole-4-carboxylic acid
Using Deprotection and Hydrolysis General Procedure followed by purification via reverse phase préparative chromatography (C18, 25 mM NH4HCO3 in water : MeCN) starting from the product from Step C, the desired product was obtained.
HRMS-ESI (m/z): [M+H]+ calcd for C22H25N6O4S2: 501.1373, found 501.136.
Example 133:2-[[6-(l,3-Benzothiazol-2-yIamino)-5-methyl-pyridazin-3-yI]-(5-hydroxy-415 methoxy-pentyl)amino]thiazole-4-carboxylic acid
Step A: methyl2-[tert-butoxycarbonyl-[5-[tert-butyl(dimethyl)silyl]oxy-4-methoxy· pentyl]amino]thiazole-4-carboxylate
Using Mitsunobu General Procedure starting from 571 mg of methyl 2-(tert20 butoxycarbonylamino)thiazole-4-carboxylate (2.21 mmol, 1.1 eq.) and 500 mg of Préparation 2b (2.01, 1 eq.) as the appropriate alcohol, 818 mg (83%) of the desired product was obtained. Ή NMR (400 MHz, DMSO-dô) 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,
387
9H), -0.01 (s, 6H); 13C NMR (100 MHz, DMSO-d6) 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]+ calcd for C22H4iN2O6SSi: 489.2, found 489.3.
Step B: methyl 2-[[5-[tert-butyl(dimethyl)silyl]oxy-4-methoxy-pentyl]amino]thiazole-4carboxylate
Using Deprotection with HFIP General 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.
XH NMR (400 MHz, DMSO-î/6) 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); 13C NMR (100 MHz, DMSO4) 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]+ calcd for Ci7H33N2O4SSi: 389.2, found 389.3.
Step C: methyl 2-[[5-[tert-butyl(dimethyl)silyl]oxy-4-methoxy-pentyl]-[5-methyl-6-[(Z)-[3(2-trimethylsilylethoxymethyl)-l,3-benzothiazol-2-ylidene]amino]pyridazin-3yl]amino]thiazole-4-carboxylate
Using Buchwald General Procedure II starting from 224 mg of the product from Step B (0.58 mmol, 1 eq.) and 349 mg of Préparation 4a (0.86 mmol, 1.5 eq.) as the appropriate halide, 357 mg (82%) of the desired product was obtained.
Ή NMR (500 MHz, DMSO-ώ) 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); 13C NMR (125 MHz, DMSO-î/6) 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; HRMS-ESI (m/z): [M+H]+ calcd for C35H55N6O5S2Si2: 759.3208, found 759.3212.
Step D: 2-[[6-(l,3-benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-(5-hydroxy-4-methoxypentyl)amino]thiazole-4-carboxylic acid
Using Deprotection and Hydrolysis General Procedure followed by purification via reverse phase préparative chromatography (C18, 25 mM NH4HCO3 in water : MeCN) starting from the product from Step C, the desired product was obtained.
HRMS-ESI (m/z): [M+H]+ calcd for C22H25N6O4S2: 501.1373, found 501.1366.
388
Example 134:2-[[6-(l,3-Benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-(4hydroxybutyl)amino]-5-[3-[4-[3-(dimethylamino)prop-l-ynyl]-2-fluorophenoxy] propyl] thiazole-4-carboxylie acid
Using Deprotection and Hydrolysis General Procedure starting from the appropriate methyl ester Préparation 5d, Step A, the desired product was obtained.
HRMS-ESI (m/z): [M+H]+ calcd for C34H37FN7O4S2: 690.2327, found 690.2347.
Example 135: 2-({6-[(l,3-Benzothiazol-2-yl)amino]-5-methylpyridazin-3yl}(methyl)amino)-5-{l-[(terAbutoxy)carbonyl]azetidin-3-yl}-l,3-thiazole-4-carboxylic acid
Step A: ethyl 2-({6-[(l,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)5-{l-[(tert-butoxy)carbonyl]azetidin-3-yl}-l,3-thiazole-4-carboxylate
To a solution of the product from Préparation 51, Step A (122 mg, 0.17 mmol, 1 eq) in 15 tetrahydrofuran (6 mL) was added tetrabutylammonium fluoride (IM in tetrahydrofuran; 0.24 mL, 1.38 mmol, 8 eq) and the mixture was heated at 50 °C for 13 h. The reaction was allowed to cool to ambient température then partitioned between dichloromethane and saturated aqueous
389
P sodium bicarbonate, and the organic phase was washed with brine, dried (magnésium sulfate), and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 24 g RediSep™ silica cartridge) eluting with a gradient of 0 - 10% methanol in dichloromethane afforded the desired product as a yellow glass (38 mg, 0.07 mmol, 38%).
LC/MS (C27H31N7O4S2) 582 [M+H]+; RT 1.26 (LCMS-V-B1)
Ή NMR (400 MHz, DMSO-d6) 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-[(l,3-benzothiazol-2-yl)aniino]-5-methylpyridazin-3-yl}(methyl)amino)-5-{l10 [(tert-butoxy)carbonyl]azetidin-3-yl}-l,3-thiazole-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 2M aqueous sodium hydroxide (0.5 mL, 1 mmol, 15 eq) and the mixture was heated at 100 °C for 30 min. The reaction was allowed to cool to ambient température and concentrated in vacuo. Purification by reverse phase automated flash chromatography (CombiFlash Rf, C18 15 13g RediSep column) eluting with a gradient of 5 - 95% acetonitrile in water afforded the desired product as a yellow solid (21.4 mg, 0.04 mmol, 59%).
HRMS-ESI (m/z) [M-H]- calcd for C25H26N7O4S2: 552.1493, found 552.1511.
Example 136: 5-(l-Benzoylazetidm-3-yI)-2-({6-[(l,3-benzothiazol-2-yl)amino]-520 methylpyridazin-3-yl}(methyl)amino)-l,3-thiazole-4-carboxylic acid
Step A: ethyl5-(l-benzoylazetidin-3-yl)-2-[methyl(5-methyl-6-{[(2Z)-3-{[2(trimethylsilyl)ethoxy]methyl}-2,3-dihydro-l,3-benzothiazol-2-ylidene]amino}pyridazin-3yl)amino]-l,3-thiazole-4-carboxylate
To a solution of the product from Préparation 51 (29.6 mg, 0.05 mmol, 1 eq) and benzoyl chloride (6.74 pL, 0.06 mmol, 1.2 eq) in dichloromethane (3 mL) was added triethylamine (0.01
390 mL, 0.1 mmol, 2 eq) and the mixture was stirred at ambient température for 3.5 h. Purification by automated flash column chromatography (CombiFlash Rf, 12 g RediSep™ silica cartridge) eluting with a gradient of 0 - 85% ethyl acetate in Ao-heptane afforded the desired product as a colourless glass (32 mg, 0.04 mmol, 92%).
LC/MS (C35H4iN7O4SiS2) 716 [M+H]+; RT 1.59 (LCMS-V-B1)
Ή NMR (400 MHz, DMSO-d6) δ 7.86 (dd, J = 7.6, 1.1 Hz, 1H), 7.75 - 7.67 (m, 3H), 7.58 7.39 (m, 5H), 7.29 - 7.22 (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-(l-benzoylazetidin-3-yl)-2-({6-[(l,3-benzothiazol-2-yl)amino]-5methylpyridazin-3-yl}(methyl)amino)-l,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 température for 5 h. The reaction was partitioned between dichloromethane and saturated aqueous sodium bicarbonate, and the organic phase was washed with saturated aqueous sodium bicarbonate then brine, dried (magnésium sulfate), and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 12 g RediSep™ silica cartridge) eluting with a gradient of 0 - 5% methanol in dichloromethane afforded the desired product as a yellow glass (17.5 mg, 0.03 mmol, 67%).
LC/MS (C29H27N7O3S2) 586 [M+H]+; RT 1.30 (LCMS-V-B1)
Ή NMR (400 MHz, DMSO-d6) 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-(l-benzoylazetidin-3-yl)-2-({6-[(l,3-benzothiazol-2-yl)amino]-5-methylpyridazin3-yl}(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 2M aqueous sodium hydroxide (0.5 mL, 1 mmol, 33.5 eq) and the mixture was heated at reflux for 2 h. The reaction was allowed to cool to ambient température and purification by reverse phase automated flash chromatography (CombiFlash Rf, C18 13g RediSep column)
391 eluting with a gradient of 5 - 95% acetonitrile in water afforded the desired product as a yellow solid (16 mg, 0.03 mmol, 96%) [as a sodium sait].
HRMS-ESI (m/z) [M-H]- calcd for C27H22N7O3S2: 556.1231, found 556.1260.
Example 137: 2-({6-[(l,3-Benzothiazol-2-yl)amino]-5-methylpyridazin-35 yI}(methyI)amino)-5-(3-{4-[3-(dimethylamino)prop-l-yn-l-yl]-2-fluorophenoxy}-2,2dimethylpropyl)-l,3-thiazole-4-carboxyIic acid
Step A: methyl5-(3f4-[3-(dimethylamino)prop-l-yn-1 -yl]-2-fluorophenoxy}-2,2dimethylpropyl) -2-[methyl(5-methyl-6-{[ (2Z) -3-{[2-(trimethylsilyl) ethoxy]methyl}-2,310 dihydro-l,3-benzothiazol-2-ylidene]amino}pyridazin-3-yl)amino]-l,3-thiazole-4-carboxylate
To a solution of the product from Préparation 3zc (8 mg, 0.02 mmol, 1 eq) in 1,4-dioxane (2 mL) was added the product from Préparation 4a (9.76 mg, 0.02 mmol, 1.3 eq), Xantphos (2.14 mg, 3.69 pmol, 0.2 eq), césium carbonate (18 mg, 0.06 mmol, 3 eq) and N,Ndiisopropylethylamine (9.64 pL, 0.06 mmol, 3 eq). The mixture was sparged with nitrogen (10 15 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 was partitioned between ethyl acetate and brine, and the organic phase was dried (magnésium sulfate) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 4 g RediSep™ silica cartridge) eluting with a gradient of 0 - 6% methanol in dichloromethane afforded the desired 20 product as a yellow solid (15 mg, 0.02 mmol, 100%).
LC/MS (C4oH50FN704SiS2) 804 [M+H]+; RT 2.65 (LCMS-V-C)
392 φ ΧΗ NMR (400 MHz, DMSO-d6) δ 7.83 (d, J = 7.9 Hz, 1H), 7.67 (d, J = 1.2 Hz, 1H), 7.49 7.42 (m, 2H), 7.35 - 7.23 (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-[(l,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)-5-(3{4-[3-(dimethylamino)prop-l -yn-1 -yl]-2-fluorophenoxy}-2,2-dimethylpropyl) -1,3-thiazole-4carboxylic 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 pL, 0.22 mmol, 3 eq) and tetrabutylammonium fluoride 10 (IM in tetrahydrofuran, 224 pL, 0.22 mmol, 3 eq) and the mixture was heated at 70 °C overnight. The solvent was removed in vacuo and purification by reverse phase automated flash chromatography (CombiFlash Rf, C18 13g RediSep column) eluting with a gradient of 5 - 95% acetonitrile in water afforded a solid that was triturated with diethyl ether, filtered, and dried under vacuum to afford the desired product as a yellow solid (13.1 mg, 0.02 mmol, 27%).
HRMS-ESI (m/z) [M+H]+ calcd for C33H35FN7O3S2: 660.2221, found 660.2264.
Example 138: 2-({6-[(l,3-Benzothiazol-2-yl)ammo]-5-methylpyridazin-3yl}(methyl)amino)-5-[l-(2-phenylacetyl)azetidin-3-yl]-l,3-thiazole-4-carboxylic acid
Step A: ethyl2-[methyl(5-methyl-6-{[(2Z)-3-{[2-(trimethylsilyl)ethoxy]methyl}-2,3-dihydro20 l,3-benzothiazol-2-ylidene]amino}pyridazin-3-yl)amino]-5-[l-(2-phenylacetyl)azetidin-3yl]-l,3-thiazole-4-carboxylate
To a solution of the product from Préparation 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 température for 3 h. The reaction 25 was diluted with dichloromethane then washed with water and brine, dried (magnésium sulfate),
393 and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 12 g RediSep™ silica cartridge) eluting with a gradient of 0 - 100% ethyl acetate in zso-heptane afforded the desired product as a yellow foam (66 mg, 0.09 mmol, 53%).
LC/MS (C36H43N7O4SiS2) 730 [M+H]+; RT 1.45 (LCMS-V-B1) ’H NMR (400 MHz, DMSO-d6) 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-[(l,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)5-[l-(2-phenylacetyl)azetidin-3-yl]-l,3-thiazole-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 température overnight. The reaction was partitioned between dichloromethane and saturated aqueous sodium bicarbonate, washed with saturated aqueous sodium bicarbonate then brine, dried (magnésium sulfate), and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 12 g RediSep™ silica cartridge) eluting with a gradient of 0 - 5% methanol in dichloromethane afforded the desired product as a yellow glass (39 mg, 0.07 mmol, 72%).
LC/MS (C30H29N7O3S2) 600 [M+H]+; RT 1.33 (LCMS-V-B1) ‘H NMR (400 MHz, DMSO-d6) 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-[(l,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)-5-[l(2-phenylacetyl)azetidin-3-yl]-l,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 a 2M aqueous sodium hydroxide (1 mL, 2 mmol, 30.8 eq) and the mixture was heated at reflux for 1.5 h. The solvent was removed in vacuo and purification by reverse phase automated flash chromatography (CombiFlash Rf, C18 13g RediSep column) eluting with a gradient of 5
394
- 95% acetonitrile in water afforded the desired product as a yellow solid (28.7 mg, 0.05 mmol, 77%) [as a sodium sait].
HRMS-ESI (m/z) [M-H]- calcd for C28H24N7O3S2: 570.1388, found 570.1423.
Example 139: 5-(Azetidin-3-yl)-2-({6-[(l,3-benzothiazol-2-yl)amino]-5-methylpyridazin5 3-yl}(methyl)amino)-l,3-thiazole-4-carboxylic acid
During the préparation of Example 138, Step C, sodium 5-(azetidin-3-yl)-2-({6-[(l,3benzothiazol-2-y l)amino] -5 -methy lpyridazin-3 -yl} (methy l)amino)-1,3-thiazole-4-carboxylate (7.5 mg, 0.02 mmol, 25%) was isolated as a byproduct [as a sodium sait].
HRMS-ESI (m/z) [M-H]- calcd for C20H18N7O2S2: 452.0969, found 452.0977.
Example 140: 2-({6-[(l,3-Benzothiazol-2-yl)amino]-5-methylpyridazin-3yl}(methyl)amino)-5-{[l-({4-[3-(dimethylamino)prop-l-yn-l-yl]-2fluorophenoxy}methyl)cyclopropyl]methyl}-l,3-thiazole-4-carboxylic acid
Step A: methyl 5-{[l-({4-[3-(dimethylamino)prop-l-yn-l-yl]-2fluorophenoxy}methyl)cyclopropyl]methyl}-2-[methyl(5-methyl-6-{[(2Z)-3-{[220487
395 (trimethylsilyl)ethoxy]methyl}-2,3-dihydro-l,3-benzothiazol-2-ylidene]amino}pyridazin-3yl)amino]-l,3-thiazole-4-carboxylate
To a solution of the product from Préparation 3zd (84 mg, 0.19 mmol, 1 eq) in 1,4-dioxane (6 mL) was added the product from Préparation 4a (103 mg, 0.25 mmol, 1.3 eq), Xantphos (22.5 mg, 0.04 mmol, 0.2 eq), césium carbonate (190 mg, 0.58 mmol, 3 eq) and N,Ndiisopropylethylamine (102 pL, 0.58 mmol, 3 eq). The mixture was sparged with nitrogen (10 min), then trîs(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 was partitioned between ethyl acetate and brine, and the organic phase was dried (magnésium sulfate), and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 4 g RediSep™ silica cartridge) eluting with a gradient of 0 - 5% methanol in dichloromethane afforded the desired product as a brown solid (98 mg, 0.12 mmol, 63%).
LC/MS (C4oH48FN704SiS2) 802 [M+H]+; RT 2.63 (LCMS-V-C) *H NMR (400 MHz, DMSO-d6) 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-[(l,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3yl}(methyl)amino)-5-{[l-({4-[3-(dimethylamino)prop-l -yn-1 -yl]-2fluorophenoxy}methyl)cyclopropyl]methyl}-l,3-thiazole-4-carboxylate
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 pL, 0.37 mmol, 3 eq) and tetrabutylammonium fluoride (IM in tetrahydrofuran; 224 pL, 0.37 mL, 0.37 mmol, 3 eq) and the mixture was heated at 70 °C overnight. The reaction was 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 13g RediSep column) eluting with a gradient of 5 - 95% acetonitrile in water afforded the desired product as a yellow solid (5.01 mg, 0.01 mmol, 6%).
LC/MS (C34H34FN7O3S2) 672 [M+H]+; RT 2.04 (LCMS-V-C)
Hl NMR (400 MHz, DMSO-d6) 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),
396 φ 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-[(l,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)atnino)-5-{[l({4-[3-(dimethylamino)prop-l-yn-l-yl]-2-fluorophenoxy}methyl)cyclopropyl]methyl}-l,35 thiazole-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 at 110 °C overnight. The reaction was concentrated in vacuo and the residue was triturated with water, filtered, and dried under vacuum to afford the desired product as a yellow solid (23.8 10 mg, 0.04 mmol, 70%) [as a lithium sait].
HRMS-ESI (m/z) [M+H]+ calcd for C33H33FN7O3S2: 658.2065, found 658.2109.
Example 141:2-[[6-(l,3-Benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yI]-(5hydroxypentyl)amino]-5-[3-[4-[3-(dimethylamino)prop-l-ynyl]-2-fluorophenoxy] propyl] thiazole-4-carboxylic acid
Using Deprotection and Hydrolysis General Procedure followed by repurification via reverse phase préparative chromatography (C18, 25 mM NH4HCO3 in water : MeCN), starting from Préparation 5f the desired product was obtained.
HRMS-ESI (m/z): [M+H]+ calcd for C35H39FN7O4S2: 704.2489 found: 704.2483.
397
Example 142:2-[[6-(l,3-Benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-[5-[2(dimethyIamino)ethyl-methyl-amino]-4-hydroxy-pentyl]amino]thiazole-4-carboxylic acid
Step A: methyl2-[tert-butoxycarbonyl-[4-[tert-butyl(dimethyl)silyl]oxy-5-[2(dimethylamino)ethyl-methyl-amino]pentyl]amino]thiazole-4-carboxylate
Using Mitsunobu General Procedure starting from 426 mg of methyl 2-(tertbutoxycarbonylamino)thiazole-4-carboxylate (1.65 mmol, 1.1 eq.) and 478 mg of Préparation 2f (1.65 mmol, 1 eq.) as the appropriate alcohol, 359 mg (43%) of the desired product was 10 obtained.
Ή NMR (400 MHz, DMSO-dô) 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]+ calcd for C26H5iN4O5SSi: 559.3, found 559.4.
Step B: methyl 2-[[4-[tert-butyl(dimethyl)silyl]oxy-5-[2-(dimethylamino)ethyl-methylamino]pentyl]amino]thiazole-4-carboxylate
Using Deprotection with HFIP General 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.
Ή NMR (500 MHz, DMSO-dô) 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); 13C NMR (125 MHz, DMSO-dô) 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,
398 φ 26.3, 24.9, -3.8, -4.3; LC-MS-ESI (m/z): [M+H]+ calcd for C2iH43N4O3SSi: 459.3, found 459.4.
Step C: methyl 2-[[4-[tert-butyl(dimethyl)silyl]oxy-5-[2-(dimethylamino)ethyl-methylamino]pentyl]-[5-methyl-6-[(Z)-[3-(2-trimethylsilylethoxymethyl)-l,3-benzothiazol-25 ylidene]amino]pyridazin-3-yl]amino]thiazole-4-carboxylate
Using Buchwald General Procedure II starting from 255 mg of the product from Step B (0.56 mmol, 1 eq.) and 226 mg of Préparation 4a (0.77 mmol, 1.0 eq.) as the appropriate halide, 311 mg (67%) of the desired product was obtained.
1H NMR (500 MHz, DMSO-d6) δ ppm 7.97 (s, 1H), 7.83 (dd, 1H), 7.70 (s, 1H), 7.46 (dd, 1H), 10 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); 13C NMR (125 MHz, DMSO-d6) 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;
HRMS-ESI (m/z): [M+H]+ calcd for C39H65N8O4S2Si2: 829.4103, found 829.4096.
Step D: 2-[[6-(l,3-benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-[5-[2(dimethylamino)ethyl-methyl-amino]-4-hydroxy-pentyl]amino]thiazole-4-carboxylic acid
Using Deprotection and Hydrolysis General Procedure followed by purification via reverse phase préparative chromatography (Cl8, 25 mM NH4HCO3 in water : MeCN) starting from the 20 product from Step C, the desired product was obtained.
HRMS-ESI (m/z): [M+H]+ calcd for C26H34N8O3S2: 571.2268, found 571.2270.
Example 143:2-[[6-(l,3-Benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-[5(dimethylamino) -4-hy droxy-penty 1] amino] thiazole-4-carboxylic acid
399
Step A: methyl2-[tert-butoxycarbonyl-[4-[tert-butyl(dimethyl)silyl]oxy-5(dimethylamino)pentyl]amino]thiazole-4-carboxylate
Using Mitsunobu General Procedure starting from 568 mg of methyl 2-(tertbutoxycarbonylamino)thiazole-4-carboxylate (2.2 mmol, 1.1 eq.) and 523 mg of Préparation 2d (2.0 mmol, 1 eq.) as the appropriate alcohol, 530 mg (53%) of the desired product was obtained.
XH NMR (500 MHz, DMSO-ife) 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); 13C NMR (125 MHz, DMSO-d6) 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; HRMS-ESI (m/z): [M+H]+ calcd for C23H44N3O5SSÊ 502.2765, found 502.2762.
Step B: methyl 2-[[4-[tert-butyl(dimethyl)silyl]oxy-5-(dimethylamino)pentyl]amino]thiazole4-carboxylate
Using Deprotection with HFIP General 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.
*H NMR (500 MHz, DMSO-tfe) 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); 13C NMR (125 MHz, DMSO-î/6) 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; HRMS-ESI (m/z): [M+H]+ calcd for Ci8H36N3O3SSi: 402.2241, found 402.2249.
Step C: methyl 2-[[4-[tert-butyl(dimethyl)silyl]oxy-5~(dimethylamino)pentyl]-[5-methyl-6[(Z)-[3-(2-trimethylsilylethoxymethyl)-l,3-benzothiazol-2-ylidene]amino]pyridazin-3yl]amino]thiazole-4-carboxylate
Using Buchwald General Procedure II starting from 72 mg of the product from Step B (0.18 mmol, 1 eq.) and 73 mg of Préparation 4a (0.18 mmol, 1.5 eq.) as the appropriate halide, 126
400 mg (91%) of the desired product was obtained.
1H NMR (500 MHz, DMSO-d6) δ ppm 7.86-7.23 (4H), 7.97 (s, 1 H), 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, 5 6H), -0.12 (s, 9H); 13C NMR (125 MHz, DMSO-ώ) 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; HRMS-ESI (m/z): [M+H]+ calcd for C36H57N7O4S2Si2: 772.3525, found 772.3521.
Step D: 2-[[6-(l,3-benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-[5-(dimethylamino)-4hydroxy-pentyl]amino]thiazole-4-carboxylic acid
Using Deprotection and Hydrolysis General Procedure followed by purification via reverse phase préparative chromatography (C18, 25 mM NH4HCO3 in water : MeCN) starting from the product from Step C, the desired product was obtained.
HRMS-ESI (m/z): [M+H]+ calcd for C23H28/N7O3S2: 514.1690, found 514.1694.
Example 144:2-iï6-(1.3-Benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-[4-hydroxy-515 (trimethylammonio)pentyl]amino]-5-[3-[2-fluoro-4-[3-(methylamino)prop-lynyl] phenoxy] propy 1] thiazole-4-carboxylate
Step A: methyl5-[3-[4-[3-[tert-butoxycarbonyl(methyl) amino]prop-l-ynyl]-2-fluorophenoxy]propyl]-2-[[4-[tert-butyl(diphenyl)silyl]oxy-5-(dimethylamino)pentyl]-[5-methyl-620 [(Z)-[3-(2-trimethylsilylethoxymethyl)-l,3-benzothiazol-2-ylidene]amino]pyridazin-3yl]amino]thiazole-4-carboxylate
401 ^b Using Alkylation with tosylate General Procedure starting from Préparation 5a and Nmethylmethanamine as the appropriate amine, the desired product was obtained.
HRMS-ESI (m/z): [M+H]+ calcd for C64H84FN8O7S2Si2: 1215.5421, found 1215.5389.
Step B: 2-[[6-(l,3-benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-[4-hydroxy-55 (trimethylammonio)pentyl]amino]-5-[3-[2-fluoro-4-[3-(methylamino)prop-lynyl]phenoxy]propyl]thiazole-4-carboxylate
The product from Step A was dissolved in the mixture of acetonitrile (4 mL/mmol) and N,Ndimethylformamide (1 mL/mmol) then iodomethane (5 eq.) was added and stirred at rt until full conversion was observed (ca. 1 h). The reaction mixture was concentrated. The crude mixture 10 which contained [5-[[5-[3-[4-[3-[tert-butoxycarbonyl(methyl)amino]prop-l-ynyl]-2-fluorophenoxy]propyl]-4-methoxycarbonyl-thiazol-2-yl]-[5-methyl-6-[(Z)-[3-(2trimethylsilylethoxymethyl)-l,3-benzothiazol-2-ylidene]amino]pyridazin-3-yl] amino]-2-[tertbutyl(diphenyl)silyl]oxy-pentyl]-trimethyl-ammonium (LC-MS-ESI (m/z): [M]+ calcd for C65H86FN8O7S2Si2: 1229.6, found 1229.4) was transferred to the next reaction using 15 Quaternary sait deprotection General Procedure, to afford the desired product.
HRMS-ESI (m/z): [M+H]+ calcd for C37H44FN8O4S2: 747.2905, found 747.2900.
Example 145: 2-({6-[(l,3-Benzothiazol-2-yl)amino]-5-methylpyridazin-3yl}(methyl)amino)-5-(l-benzylazetidin-3-yl)-l,3-thiazole-4-carboxylic acid
Step A: ethyl5-(l-benzylazetidin-3-yl)-2-[methyl(5-methyl-6-{[(2Z)-3-{[2(trimethylsilyl)ethoxy]methyl}-2,3-dihydro-l,3-benzothiazol-2-ylidene]amino}pyridazin-3yl)amino]-l,3-thiazole-4-carboxylate
To a solution of the product from Préparation 51 (36.3 mg, 0.06 mmol, 1 eq) in acetonitrile (3 mL) was added benzaldehyde (12.1 pL, 0.12 mmol, 2 eq), sodium triacetoxy borohydride (37.7 25 mg, 0.18 mmol, 3 eq) and glacial acetic acid (two drops) and the mixture was stirred at ambient
402 température overnight. The reaction was diluted with ethyl acetate and washed with saturated aqueous sodium bicarbonate then brine, dried (magnésium sulfate), and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 12 g RediSep™ silica cartridge) eluting with a gradient of 0 - 70% ethyl acetate in zso-heptane afforded the desired product as a colourless gum (28.6 mg, 0.04 mmol, 69%).
LC/MS (C35H43N7O3SiS2) 702 [M+H]+; RT 1.49 (LCMS-V-B1)
Ή NMR (400 MHz, DMSO-d6) 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), 3.77 - 3.65 (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-[(l,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)5-(l -benzylazetidin -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 température overnight. The reaction was partitioned between dichloromethane and saturated aqueous sodium bicarbonate, and the organic phase was washed with saturated aqueous sodium bicarbonate then brine, dried (magnésium sulfate), and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 4 g RediSep™ silica cartridge) eluting with a gradient of 0 - 5% methanol in dichloromethane afforded the desired product as a yellow glass (17 mg, 0.03 mmol, 73%).
LC/MS (C29H29N7O2S2) 572 [M+H]+; RT 1.14 (LCMS-V-B1) *H NMR (400 MHz, DMSO-d6) 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-[(l,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)-5-(lbenzylazetidin-3-yl)-l,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 2M aqueous sodium hydroxide (0.5 mL, 1 mmol, 33.6 eq) and the mixture was heated at reflux for 2 h. The reaction was allowed to cool to ambient température and the solvent was removed in vacuo. Purification by reverse phase automated flash chromatography (CombiFlash
403
Rf, C18 13g RediSep column) eluting with a gradient of 5 - 95% acetonitrile in water afforded the desired product as a yellow solid (13.9 mg, 0.03 mmol, 86%) [as a sodium sait].
HRMS-ESI (m/z) [M+H]+ calcd for C27H26N7O2S2: 544.1584, found 544.1616.
Example 146:3-[[5-[[6-(l,3-Benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-[45 carboxy-5- [3- [2-fluoro-4- [3-(methylamino)prop-l-ynyl]phenoxy] propyl] thiazol-2yl]amino]-2-hydroxy-pentyl]-dimethyl-ammonio]propane-l-suIfonate
Example 144, Step A was suspended in MeCN (5 mL/mmol) then oxathiolane 2,2-dioxide (10 eq.) was added and stirred at 60°C for on (full conversion was observed). The reaction mixture 10 was concentrated. The crude mixture which contained 3-[[5-[[5-[3-[4-[3-[tertbutoxycarbony l(methyl)amino]prop-1-ynyl]-2-fluoro-phenoxy]propyl]-4-methoxycarbonylthiazol-2-yl]-[5-methyl-6-[(Z)-[3-(2-trimethylsilylethoxymethyl)-l,3-benzothiazol-2ylidene]amino]pyridazin-3-yl]amino] -2-[tert-butyl(diphenyl)silyl] oxy-pentyl]-dimethylammonio]propane-l-sulfonate (LC-MS-ESI (m/z): [M+H]+ calcd for C67H9oFN80ioS3SÎ2: 15 1337.5, found 1337.6) was transferred directly to the next reaction using Quaternary sait deprotection General Procedure, to afford the desired product.
HRMS-ESI (m/z): [M+H]+ calcd for C39H48FN8O7S3: 855.2787, found 855.2786.
Example 147:2-({6-[(l,3-Benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(4hydroxybutyl)amino)-5-[3-(4-{3-[bis(3-sulfopropyl)amino]prop-l-yn-l-yl}-220 fluorophenoxy)propyl]-l,3-thiazole-4-carboxylic acid
404
Example 147 was isolated as the side product from the reaction of Example 148, Step B.
HRMS-ESI (m/z): [M+H]+ calcd for C38H44FN7O10S4: 906.2089, found 906.2098.
Example 148:2-rr6-(l.3-Benzothiazol-2-vlamino)-5-methyl-pyridazin-3-vl1-(4hydroxybutyI)amino]-5-[3-[2-fluoro-4-[3-(3-sulfopropylamino)prop-lynyl] phenoxy] propyl] thiazole-4-carboxylic acid
Step A: 5-[3-[4-(3-aminoprop-l-ynyl)-2-fluoro-phenoxy]propyl]-2-[[6-(l,3-benzothiazol-210 ylamino)-5-methyl-pyridazin-3-yl]-(4-hydroxybutyl)amino]thiazole-4-carboxylic acid
Using Deprotection and Hydrolysis General Procedure starting from 370 mg of Préparation 5i, Step A (0.36 mmol, 1 eq.), 150 mg (62%) of the desired product was obtained.
Ht NMR (500 MHz, DMSO-d6) 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), 15 3.47 (t, 2H), 3.27 (t, 2H), 2.47 (s, 3H), 2.14 (m, 2H), 1.74 (m, 2H), 1.52 (m, 2H); 13C NMR (125 MHz, DMSO-dô) δ ppm 126.7, 126.5,122.6,122.2, 119.1,118.6, 116.8, 115.9, 91.0, 81.1,
405 φ 68.8, 60.9, 47.2, 31.6, 31.0, 29.9, 24.2, 23.2, 17.7; HRMS-ESI (m/z): [M+HJ+ calcd for C32H33FN7O4S2: 662.2014, found 662.2011.
Step B: 2-[[6-(l,3-benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-(4hydroxybutyl)amino]-5-[3-[2-fluoro-4-[3-(3-sulfopropylamino)prop-l5 ynyl]phenoxy]propyl]thiazole-4-carboxylic acid
To a suspension of the product from Step A in dry acetonitrile oxathiolane 2,2-dioxide was added in one portion. The resulting mixture was stirred overnight at rt, where the disubstituted product also had formed (Example 147). The crude product was dissolved in DMSO, and then it was purified via reverse phase préparative chromatography (25 mM NH4HCO3 in water : 10 MeCN) to obtain the desired product.
HRMS-ESI (m/z): [M+H]+ calcd for C35H39FN7O7S3: 784.2052, found 784.2052.
Example 149: 2-({6-[(l,3-Benzothiazol-2-yI)ammo]-5-methylpyridazm-3yl}(methyI)amino)-5-(l-methanesulfonylazetidin-3-yl)-l,3-thiazole-4-carboxylic acid
Step A: ethyl5-(l-methanesulfonylazetidin-3-yl)-2-[methyl(5-methyl-6-{[(2Z)-3-{[2(trimethylsilyl)ethoxy]methyl}-2,3-dihydro-l,3-benzothiazol-2-ylidene]amino}pyridazin-3yl)amino]-l,3-thiazole-4-carboxylate
To a solution of the product from Préparation 51 (24.6 mg, 0.04 mmol, 1 eq) in dichloromethane (3 mL) was added methanesulfonyl chloride (3.73 pL, 0.05 mmol, 1.2 eq), followed by a 20 catalytic amount of 4-(dimethylamino)pyridine and triethylamine (0.01 mL, 0.08 mmol, 2 eq) and the mixture was stirred at ambient température overnight. The reaction was partitioned between dichloromethane and saturated aqueous sodium bicarbonate, and the organic phase was washed with brine, dried (magnésium sulfate), and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 4 g RediSep™ silica cartridge)
406 eluting with a gradient of 0 - 100% ethyl acetate in Ao-heptane afforded the desired product as an off-white solid (16.4 mg, 0.02 mmol, 59%).
LC/MS (C29H39N7O5SiS3) 690 [M+H]+; RT 1.41 (LCMS-V-B1) XH NMR (400 MHz, DMSO-d6) δ 7.87 (dd, J = 7.6,1.1 Hz, 1H), 7.71 (d, J = 1.1 Hz, 1H), 7.51 5 - 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-[(l,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)10 5-(l-methanesulfonylazetidin-3-yl)-l,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 température overnight. The reaction was diluted with dichloromethane, washed with saturated aqueous sodium bicarbonate (x2) then brine, dried (magnésium sulfate), and 15 concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash
Rf, 4 g RediSep™ silica cartridge) eluting with a gradient of 0 - 10% methanol in dichloromethane afforded the desired product as a beige solid (10 mg, 0.02 mmol, 75%).
LC/MS (C23H25N7O4S3) 560 [M+H]+; RT 1.24 (LCMS-V-B1)
Ή NMR (400 MHz, DMSO-d6) δ 7.92 (br s, 1H), 7.72 (s, 1H), 7.51 - 7.42 (m, 1H), 7.40 20 7.34 (m, 1H), 7.27 - 7.15 (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-[(l,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)-5-(lmethanesulfonylazetidin-3-yl)-l,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 2M aqueous sodium hydroxide (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 13g RediSep column) eluting with a gradient of 5 - 95% acetonitrile in water afforded the desired product as a yellow solid (7.6 mg, 0.01 mmol, 80%) [as a sodium sait].
HRMS-ESI (m/z) [M+H]+ calcd for C21H22N7O4S3: 532.0890, found 532.0894.
407
Example 150:2-[[6-(l,3-Benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-[4-hydroxy-5(methylamino)pentyl]amino] -5-[3-[4- [3-(dimethylamino)prop-l-ynyl] -2-fluorophenoxy] propyl] thiazole-4-carboxylic acid
Using Alkylation with tosylate General Procedure starting from Préparation 5b and methanamine as the appropriate amine, intermediate methyl 2-[[4-[tertbutyl(diphenyl)silyl]oxy-5-(methylamino)pentyl]-[5-methyl-6-[(Z)-[3-(2- trimethylsilylethoxymethyl)-l,3-benzothiazol-2-ylidene] amino]pyridazin-3-yl] amino]-5-[3-[4[3-(dimethylamino)prop-l -ynyl]-2-fluoro-phenoxy]propyl]thiazole-4-carboxylate was not 10 isolated but after the volatiles was evaporated Deprotection and Hydrolysis General Procedure was applied to obtain the desired product.
HRMS-ESI (m/z): [M+2H]2+ calcd for C36H43FN8O4S2: 367.1411, found 367.1413.
Example 151:2-[[6-(l,3-Benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-(4hydroxybutyl)amino] -5- [3- [2-fluoro-4- [3- [methyl(3-sulfopropyl)amino]prop-l15 ynyl]phenoxy] propyl] thiazole-4-carboxylic acid
408
Step A: 3-[methyl(prop-2-ynyl)amino]propane-l-sulfonic acid
To 0.34 mL of N-methylpropargylamine (4 mmol, 1 eq.) in 1 mL of MeCN was added 244 mg of oxathiolane 2,2-dioxide (2 mmol, 0.5 eq.) and the mixture was stirred at 60 °C for 3 h. After 5 concentration, the residue was purified via flash column chromatography using DCM and
MeOH as eluents to give 299 mg (78%) of the desired product.
Ή NMR (400 MHz, DMSO-îA) 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]-410 methoxycarbonyl-thiazol-5-yl]propoxy]-3-fluoro-phenyl]prop-2-ynyl-methylamino]propane-l -sulfonic acid
Using Sonogashira General Procedure starting from the product from Step A and Préparation 3c, Step A as starting materials, 303 mg (56%) of the desired product.
‘H NMR (400 MHz, DMSO-c/6) δ ppm 12.45 (vbrs, 1H), 7. 45 (dd, 1H), 7.33 (dm, 1H), 7.18 15 (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-methoxycarbonylthiazol-5-yl]propoxy]-3-fluoro-phenyl]prop-2-ynyl-methyl-amino]propane-l-sulfonic acid
Using Deprotection with HFIP General Procedure starting from the product from Step B as the appropriate carbamate, 200 mg (81%) of the desired product was obtained.
XH NMR (500 MHz, DMSO-de) δ ppm 10.38 (s, 1H), 7.59 (t, 1H), 7.47 (dd, 1H), 7.33 (m, 1H),
409 φ 7.18 (t, 1Η), 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); 13C NMR (125 MHz, DMSO-d6) 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-[(Z)-[3-(2trimethylsilylethoxy methyl)-! ,3-benzothiazol-2-ylidene]amino]pyridazin-3-yl]amino]-4methoxycarbonyl-thiazol-5-yl]propoxy]-3-fluoro-phenyl]prop-2-ynyl-methylamino]propane-l-sulfonic acid
Using Buchwald General Procedure II starting from the product from Step C and 10 Préparation 4a as the appropriate halide, 270 mg (88%) of the desired product was obtained.
'H NMR (500 MHz, DMSO-rf6) 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, 15 2H), 0.79 (s, 9H), -0.02 (s, 6H), -0.12 (s, 9H); 13C NMR (125 MHz, DMSO-d6) 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, 26.2, 23.8, 23.1, 21.1, 17.9, 17.8, -1.0, -4.9.
Step E: 2-[[6-(l,3-benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-(4hydroxybutyl)amino]-5-[3-[2-fluoro-4-[3-[methyl(3-sulfopropyl)amino]prop-l20 ynyl]phenoxy]propyl]thiazole-4-carboxylic acid
The product from Step D and LiOHxHzO (10 eq.) in a 1:1 mixture of THF and water was kept at 50 °C for 18 h. After treating the reaction mixture with 2 mL of cc HCl at 0 °C, the reaction was kept 50 °C for 1 h. After pouring the mixture onto cold ammonia solution, the precipitated material was filtered off and purified via reverse phase préparative chromatography (25 mM 25 NH4HCO3 in water : MeCN) to give the desired product.
HRMS-ESI (m/z): [M+H]+ calcd for C36H41FN7O7S3: 798.2214 found: 798.2204.
Example 152: 5-(l-Acetylazetidin-3-yl)-2-({6-[(l,3-benzothiazol-2-yl)amino]-5methylpyridazin-3-yI}(methyl)amino)-l,3-thiazole-4-carboxylic acid
410
Step A: ethyl5-(l-acetylazetidin-3-yl)-2-[methyl(5-methyl-6-{[(2Z)-3-{[2(trimethylsilyl)ethoxy]methyl}-2,3-dihydro-l,3-benzothiazol-2-ylidene]amino}pyridazin-3yl)amino]-l,3-thiazole-4-carboxylate
To a solution of the product from Préparation 51 (100 mg, 0.16 mmol, 1 eq) in dichloromethane (5 mL) was added acetyl chloride (14 pL, 0.2 mmol, 1.2 eq) and triethylamine (0.05 mL, 0.33 mmol, 2 eq) and the mixture was stirred at ambient température for 4 h. The reaction was partitioned between dichloromethane and saturated aqueous sodium bicarbonate, and the organic phase was washed with brine, dried (magnésium sulfate), and concentrated in vacuo.
Purification by automated flash column chromatography (CombiFlash Rf, 4 g RediSep™ silica cartridge) eluting with a gradient of 0 - 100% ethyl acetate in Ao-heptane afforded the desired product as a cream foam (67.7 mg, 0.1 mmol, 63%).
LC/MS (C30H39N7O4S1S2) 654 [M+H]+; RT 1.37 (LCMS-V-B1) ‘H NMR (400 MHz, DMSO-d6) δ 7.87 (dd, J = 7.7,1.1 Hz, 1H), 7.70 (d, J = 1.2 Hz, 1H), 7.51 15 - 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-(l-acetylazetidin-3-yl)-2-({6-[(l,3-benzothiazol-2-yl)amino]-520 methylpyridazin -3 -yl} (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 (IM 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 was allowed to cool to ambient température, then partitioned between 25 dichloromethane and saturated aqueous sodium bicarbonate and the organic phase was washed with brine, dried (magnésium sulfate), and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 12 g RediSep™ silica cartridge) eluting with a
411 gradient of 0 - 5% methanol in dichloromethane afforded the desired product as a yellow glass (10.5 mg, 0.02 mmol, 19%).
LC/MS (C24H25N7O3S2) 524 [M+H]+; RT 0.97 (LCMS-V-B1)
Ή NMR (400 MHz, DMSO-d6) δ 7.94 (br s, 1H), 7.72 (s, 1H), 7.55 (br s, 1H), 7.39 (t, J = 7.5 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-(l-acetylazetidin-3-yl)-2-({6-[(l,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3yl}(methyl)amino)-l,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 IM aqueous 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 13g RediSep column) eluting with a gradient of 5 - 95% acetonitrile in water afforded the desired product as a white solid (2 mg, 4.04 pmol, 20%).
HRMS-ESI (m/z) [M+H]+ calcd for C22H22N7O3S2: 496.1220, found 496.1252.
Example 153: 2-({6-[(l,3-Benzothiazol-2-yl)amino]-5-methyIpyridazin-3yl}(methyI)amino)-5-(l-phenyIazetidin-3-yl)-l,3-thiazole-4-carboxylic acid
Step A: ethyl 2-[methyl(5-methyl-6-{[(2Z)-3-{[2-(trimethylsilyl)ethoxy]methyl}-2,3-dihydro20 l,3-benzothiazol-2-ylidene]amino}pyridazin-3-yl)amino]-5-(l-phenylazetidin-3-yl)-l,3thiazole-4-carboxylate
To a solution of the product from Préparation 51 (100 mg, 0.16 mmol, 1 eq) in 1,4-dioxane (3 mL) was added bromobenzene (20.6 pL, 0.2 mmol, 1.2 eq), césium carbonate (160 mg, 0.49 mmol, 3 eq), Λζ/V-diisopropylethylamine (0.08 mL, 0.49 mmol, 3 eq), Xantphos (18.9 mg, 0.03 25 mmol, 0.2 eq) and trîs(dibenzylideneacetone)dipalladium(0) (15 mg, 0.02 mmol, 0.1 eq) and
412 φ the mixture was heated at reflux overnight. The reaction was partitioned between dichloromethane and water, and the organic phase was washed with brine, dried (magnésium sulfate), and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 12 g RediSep™ silica cartridge) eluting with a gradient of 0 - 50% ethyl 5 acetate in Ao-heptane afforded the desired product as a yellow solid (44.2 mg, 0.06 mmol, 39%).
LC/MS (C34H41N7O3S1S2) 688 [M+H]+; RT 1.58 (LCMS-V-B1)
Ή NMR (400 MHz, DMSO-d6) 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, 10 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-[(l,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)5-(l -phenylazetidin-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 température overnight. The reaction was diluted with dichloromethane and washed with saturated aqueous sodium bicarbonate (x2) then brine, dried (magnésium sulfate), and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash 20 Rf, 4 g RediSep™ silica cartridge) eluting with a gradient of 0 - 5% methanol in dichloromethane afforded the desired product as a yellow glass (18 mg, 0.03 mmol, 51%).
LC/MS (C28H27N7O2S2) 558 [M+H]+; RT 1.49 (LCMS-V-B1)
Ή NMR (400 MHz, DMSO-d6) δ 7.83 (br s, 1H), 7.70 (s, 1H), 7.52 (br s, 1H), 7.39 - 7.31 (m, 1H), 7.27 - 7.16 (m, 3H), 6.78 - 6.70 (m, 1H), 6.60 - 6.51 (m, 2H), 4.82 - 4.68 (m, 1H), 4.39 25 - 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-[(l,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)-5-(lphenylazetidin-3-yl)-l,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 IM aqueous lithium hydroxide (0.16 mL, 0.16 mmol, 5 eq) and the mixture was heated 30 at reflux for 6 h. Purification by reverse phase automated flash chromatography (CombiFlash
413
Rf, C18 13g RediSep column) eluting with a gradient of 5 - 95% acetonitrile in water afforded the desired product as a yellow solid (9.5 mg, 0.02 mmol, 56%) [as a lithium sait].
HRMS-ESI (m/z) [M+H]+ calcd for C26H24N7O2S2: 530.1427, found 530.1455.
Example 154: 5-[l-(Benzenesulfonyl)azetidin-3-yl]-2-({6-[(l,3-benzothiazol-2-yl)amino]5-methylpyridazin-3-yl}(methyl)amino)-l,3-thiazole-4-carboxylic acid
Step A: ethyl5-[l-(benzenesulfonyl)azetidin-3-yl]-2-[methyl(5-methyl-6-{[(2Z)-3-{[2(trimethylsilyl)ethoxy]methyl}-2,3-dihydro-l,3-benzothiazol-2-ylidene]amino}pyridazin-3yl)amino]-l,3-thiazole-4-carboxylate
To a solution of the product from Préparation 51 (46.7 mg, 0.08 mmol, 1 eq) in dichloromethane (3 mL) was added benzenesulfonyl chloride (11.7 pL, 0.09 mmol, 1.2 eq), followed by 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 température overnight. The reaction was diluted with dichloromethane and washed with saturated aqueous sodium bicarbonate then brine, then dried (magnésium sulfate) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 4 g RediSep™ silica cartridge) eluting with a gradient of 0 67% ethyl acetate in /so-heptane afforded the desired product as a yellow glass (40.3 mg, 0.05 mmol, 70%).
LC/MS (C34H4iN7O5SiS3) 752 [M+H]+; RT 1.47 (LCMS-V-B1)
Ή NMR (400 MHz, DMSO-d6) 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-[l-(benzenesulfonyl)azetidin-3-yl]-2-({6-[(l,3-benzothiazol-2-yl)amino]-5methylpyridazin-3-yl}(methyl)amino)-l,3-thiazole-4-carboxylate
414 b 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 température overnight. The reaction was diluted with dichloromethane and washed with saturated aqueous sodium bicarbonate (x2) then brine, dried (magnésium sulfate), and 5 concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash
Rf, 4 g RediSep™ silica cartridge) eluting with a gradient of 0 - 6% methanol in dichloromethane afforded the desired product as a yellow solid (20 mg, 0.03 mmol, 60%).
LC/MS (C28H27N7O4S3) 622 [M+H]+; RT 1.37 (LCMS-V-B1)
Ή NMR (400 MHz, DMSO-d6) δ 8.00 (br s, 1H), 7.94 - 7.88 (m, 2H), 7.77 (t, J = 7.6 Hz, 2H), 10 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-[l-(benzenesulfonyl)azetidin-3-yl]-2-({6-[(l,3-benzothiazol-2-yl)amino]-5niethylpyridazin-3-yl}(methyl)amino)-l,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 15 added IM aqueous 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 15.5g Gold RediSep column) eluting with a gradient of 5 - 95% acetonitrile in water afforded the desired product as a cream solid (4.9 mg, 0.01 mmol, 26%).
HRMS-ESI (m/z) [M+H]+ calcd for C26H24N7O4S3: 594.1046, found 594.1060.
Example 155: 5-(l-Benzenesulfonamido-3-hydroxypropan-2-yl)-2-({6-[(l,3-benzothiazol2-yl)amino]-5-methylpyridazin-3-yl}(methyI)amino)-l,3-thiazole-4-carboxylic acid
5-(l-Benzenesulfonamido-3-hydroxypropan-2-yl)-2-({6-[(l,3-benzothiazol-2-yl)amino]-5methylpyridazin-3-yl}(methyl)amino)-l,3-thiazole-4-carboxylic acid (2.9 mg, 4.74 pmol, 25 15%) was isolated as a byproduct during the purification of Example 154, Step C.
415 φ HRMS-ESI (m/z) [M+H]+ calcd for C26H26N7O5S3: 612.1152, found 612.1154
Example 156: 2-({6-[(l,3-Benzothiazol-2-yl)amino]-5-methylpyridazin-3yl}(methyl)amino)-5-[l-(2-methylpropyl)azetidin-3-yl]-l,3-thiazole-4-carboxylic acid ,N S
Nx S
Step A: ethyl2-[methyl(5-methyl-6-{[(2Z)-3-{[2-(trimethylsilyl)ethoxy]methyl}-2,3-dihydrol,3-benzothiazol-2-ylidene]amino}pyridazin-3-yl)amino]-5-[l-(2-methylpropyl)azetidin-3yl]-l,3-thiazole-4-carboxylate
To a solution of the product from Préparation 51 (134 mg, 0.22 mmol, 1 eq) in 2:1 acetonitrile / dichloromethane (6 mL) was added isobutyraldéhyde (0.04 mL, 0.44 mmol, 2 eq) followed 10 by sodium triacetoxyborohydride (139 mg, 0.66 mmol, 3 eq) and glacial acetic acid (10 pL) and the mixture was stirred at ambient température for 4.5 h. The reaction was partitioned between dichloromethane and saturated aquesous sodium bicarbonate, and the organic phase was washed with brine, dried (magnésium sulfate), and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 12 g RediSep™ silica cartridge) 15 eluting with a gradient of 0 - 60% ethyl acetate in zso-heptane afforded the desired product as a cream foam (81.5 mg, 0.12 mmol, 56%).
LC/MS (C32H45N7O3SiS2) 668 [M+H]+; RT 1.25 (LCMS-V-B1) *H NMR (400 MHz, DMSO-d6) 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 20 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-[(l,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)5-[l-(2-methylpropyl)azetidin-3-yl]-l,3-thiazole-4-carboxylate
416
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 température overnight. The reaction was quenched by the addition of saturated aqueous sodium bicarbonate, then extracted with dichloromethane and the organic extract was 5 washed with brine, dried (magnésium sulfate), and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 12 g RediSep™ silica cartridge) eluting with a gradient of 0 - 5% methanol in dichloromethane afforded the desired product as a yellow solid (50 mg, 0.09 mmol, 76%).
LC/MS (C26H31N7O2S2) 538 [M+H]+; RT 0.91 (LCMS-V-B1)
Ή NMR (400 MHz, DMSO-d6) 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-[(l,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)-5-[l15 (2-methylpropyl)azetidin-3-yl]-l,3-thiazole-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 IM aqueous 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 13g RediSep column) eluting with a gradient of 5 - 95% acetonitrile in 20 water afforded the desired product as a yellow solid (34.2 mg, 0.07 mmol, 72%).
HRMS-ESI (m/z) [M+H]+ calcd for C24H28N7O2S2: 510.1740, found 510.1776.
Example 157: 2-({6-[(l,3-Benzothiazol-2-yl)amino]-5-methylpyridazin-3yl}(methyl)amino)-5-(l-methylazetidin-3-yl)-l,3-thiazole-4-carboxylic acid
417
Step A: ethyl2-[methyl(5-methyl-6-{[(2Z)-3-{[2-(trimethylsilyl)ethoxy]methyl}-2,3-dihydrol,3-benzothiazol-2-ylidene]amino}pyridazin-3-yl)amino]-5-(l~methylazetidin-3-yl)-l,3thiazole-4-carboxylate
To a solution of the product from Préparation 51 (134 mg, 0.22 mmol, 1 eq) in 2:1 acetonitrile / dichloromethane (6 mL) was added formaldéhyde (37% in water; 0.03 mL, 0.44 mmol, 2 eq) followed by sodium triacetoxyborohydride (139 mg, 0.66 mmol, 3 eq) and glacial acetic acid (10 pL) and the mixture was stirred at ambient température for 4.5 h. The reaction was diluted with dichloromethane then washed with saturated aqueous sodium bicarbonate followed by brine, dried (magnésium sulfate), and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 12 g RediSep™ silica cartridge) eluting with a gradient of 0 - 100% ethyl acetate in Ao-heptane afforded the desired product as a cream solid (65 mg, 0.1 mmol, 47%).
LC/MS (C29H39N7O3SiS2) 626 [M+H]+; RT 1.18 (LCMS-V-B1) XH NMR (400 MHz, DMSO-d6) 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-[(l,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)~ 5-(l-methylazetidin-3-yl)-l,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 température overnight. The reaction was quenched by the addition of saturated aqueous sodium bicarbonate, then extracted with dichloromethane and the organic extract was washed with brine, dried (magnésium sulfate), and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 12 g RediSep™ silica cartridge) eluting with a gradient of 0 - 10% methanol in dichloromethane afforded the desired product as a yellow solid (40 mg, 0.08 mmol, 78%).
LC/MS (C23H25N7O2S2) 496 [M+H]+; RT 0.80 (LCMS-V-B1) XH NMR (400 MHz, DMSO-d6) 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),
418 φ 3.76 (s, 3Η), 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-[(l,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)-5-(lmethylazetidin-3-yl) -l,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 IM aqueous 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 13g RediSep column) eluting with a gradient of 5 - 95% acetonitrile in water afforded the desired product as a yellow solid (15.5 mg, 0.03 mmol, 41%).
HRMS-ESI (m/z) [M+H]+ calcd for C21H22N7O2S2: 468.1271, found 468.1301.
Example 158: 2-({6-[(l,3-Benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(3carboxypropyl)amino)-l,3-thiazole-4-carboxylic acid
Step A: ethyl2-{[4-(tert-butoxy)-4-oxobutyl](5-methyl-6-{[(2Z)-3-{[215 (trimethylsilyl)ethoxy]methyl}-2,3-dihydro-l,3-benzothiazol-2-ylidene]amino}pyridazin-3yl)amino}-l,3-thiazole-4-carboxylate
To a solution of the product from Préparation 3ze (1.74 g, 5.53 mmol, 1.5 eq) in 1,4-dioxane (60 mL) was added the product from Préparation 4a (1.5 g, 3.69 mmol, 1 eq), N,Ndiisopropylethylamine (1.93 mL, 11.1 mmol, 3 eq), césium carbonate (3.6 g, 11.1 mmol, 3 20 eq) and Xantphos (427 mg, 0.74 mmol, 0.2 eq) and the mixture was sparged 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 sealed flask at 140 °C for 2 h. The reaction was allowed to cool to ambient température then partitioned between water and ethyl acetate, the aqueous phase was extracted with ethyl acetate (3 x 80 mL), and the combined organic extracts were washed with
419 brine, dried (magnésium sulfate), and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 40 g RediSep™ silica cartridge) eluting with a gradient of 0 - 25% ethyl acetate in zso-heptane afforded the desired product as a pale yellow solid (0.93 g, 1.36 mmol, 37%).
LC/MS (C32H44N6O5SiS2) 685 [M+H]+; RT 1.75 (LCMS-V-B1)
Ή NMR (400 MHz, DMSO-d6) 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-[(l,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}[4-(ethoxycarbonyl)1,3-thiazol-2-yl]amino)butanoic 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 température for 4 h. The reaction was quenched by the addition of saturated aqueous sodium bicarbonate then extracted with dichloromethane (3 x 40 mL) and the combined organic extracts were washed with brine, dried (magnésium sulfate), and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 24 g RediSep™ silica cartridge) eluting with a gradient of 0 — 20% methanol in dichloromethane afforded the desired product as a yellow solid (186.8 mg, 0.37 mmol, 28%).
LC/MS (C22H22N6O4S2) 499 [M+H]+; RT 1.27 (LCMS-V-B1) ‘H NMR (400 MHz, DMSO-d6) 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-[(l,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(3carboxypropyl) amino) -1,3-thiazole-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 heated at reflux for 2 h. The reaction was concentrated in vacuo and purification by reverse phase automated flash chromatography (CombiFlash Rf, C18 13g RediSep column) eluting with 5 95% acetonitrile in water afforded a crude product. This was dissolved in methanol and a few
420 drops of triethylamine then loaded onto a methanol-conditioned PE-ΑΧ cartridge, washed successively with methanol and dichloromethane, then eluted with 10% formic acid in dichloromethane, and concentrated in vacuo to afford the desired product as a yellow solid (28.9 mg, 0.06 mmol, 38%).
HRMS-ESI (m/z) [M+H]+ calcd for C20H19N6O4S2: 471.0904, found 471.0915.
Example 159: 2-({6-[(l,3-BenzothiazoI-2-yl)amino]-5-methylpyridazin-3yI}(methyl)amino)-5-[l-(benzylcarbamoyl)azetidin-3-yl]-l,3-thiazole-4-carboxylic acid
Step A: ethyl5-[l-(benzylcarbamoyl)azetidin-3-yl]-2-[methyl(5-methyl-6-{[(2Z)-3-{[2(trimethylsilyl)ethoxy]methyl}-2,3-dihydro-l,3-benzothiazol-2-ylidene]amino}pyridazin-3yl)amino]-l,3-thiazole-4-carboxylate
To a solution of the product from Préparation 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 triethylamine (0.07 mL, 0.49 mmol, 2 eq) and the mixture was stirred at ambient température overnight. The reaction was partitioned between dichloromethane and saturated aqueous ammonium chloride, and the organic phase was washed with brine, dried (magnésium sulfate), and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 12 g RediSep™ silica cartridge) eluting with a gradient of 0 - 100% ethyl acetate in Ao-heptane afforded the desired product as a yellow solid (125 mg, 0.17 mmol, 68%).
LC/MS (C36H44N8O4SiS2) 745 [M+H]+; RT 1.43 (LCMS-V-B1) XH NMR (400 MHz, DMSO-d6) 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 - 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).
421
Step B: ethyl2-({6-[(l,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)5-[l- (benzylcarbamoyl)azetidin -3 -y l]-1,3-thiazole-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 température overnight. The reaction was quenched by the addition of saturated aqueous sodium bicarbonate then extracted with dichloromethane, and the organic extract was washed with brine, dried (magnésium sulfate), and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 12 g RediSep™ silica cartridge) eluting with a gradient of 0 - 9% methanol in dichloromethane afforded the desired product as a yellow glass (87.6 mg, 0.14 mmol, 85%).
LC/MS (C3oH3oN803S2) 615 [M+H]+; RT 1.10 (LCMS-V-B1)
Hl NMR (400 MHz, DMSO-d6) 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-[(l,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)-5-[l(benzylcarbamoyl)azetidin-3-yl]-l,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 IM aqueous 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 13g RediSep column) eluting with a gradient of 5 - 95% acetonitrile in water afforded the desired product as a yellow solid (25.6 mg, 0.04 mmol, 31%).
HRMS-ESI (m/z) [M+H]+ calcd for C2sH27N8O3S2: 587.1642, found 587.1669.
Example 160: 2-({6-[(l,3-Benzothiazol-2-yl)amino]-5-methyIpyridazin-3yl}(methyl)amino)-5-[l-(ethylcarbamoyl)azetidin-3-yl]-l,3-thiazole-4-carboxylic acid
422
Step A: ethyl5-[l-(ethylcarbamoyl)azetidin-3-yl]-2-[methyl(5-methyl-6-{[(2Z)-3-{[2(trimethylsilyl)ethoxy]methyl}-2,3-dihydro-l,3-benzothiazol-2-ylidene]amino}pyridazin-3yl)amino]-l,3-thiazole-4-carboxylate
To a solution of the product from Préparation 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 triethylamine (0.03 mL, 0.25 mmol, 1.5 eq) and the mixture was stirred at ambient température overnight. The reaction was partitioned between dichloromethane and saturated aqueous ammonium chloride, and the organic phase was washed with brine, dried (magnésium sulfate), and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 4 g RediSep™ silica cartridge) eluting with a gradient of 0 - 100% ethyl acetate in Ao-heptane afforded the desired product as a cream solid (78 mg, 0.11 mmol, 70%).
LC/MS (C3iH42N8O4SiS2) 683 [M+H]+; RT 1.38 (LCMS-V-B1) XH NMR (400 MHz, DMSO-d6) δ 7.86 (dd, J = 7.6, 1.1 Hz, 1H), 7.69 (d, J= 1.1 Hz, 1H), 7.51 15 - 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-[(l,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)20 5-[l-(ethylcarbamoyl)azetidin-3-yl]-l,3-thiazole-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 température overnight. The reaction was quenched by the addition of saturated aqueous sodium bicarbonate and extracted with dichloromethane, and the organic extract was washed 25 with brine, dried (magnésium sulfate), and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 4 g RediSep™ silica cartridge) eluting with a
423 φ gradient of 0 - 10% methanol in dichloromethane afforded the desired product as a yellow glass (44 mg, 0.08 mmol, 70%).
LC/MS (C25H28N8O3S2) 553 [M+H]+; RT 1.00 (LCMS-V-B1) 'H NMR (400 MHz, DMSO-d6) δ 11.18 (br s, 1H), 7.93 (br s, 1H), 7.70 (s, 1H), 7.46 (br s, 5 1H), 7.43 - 7.34 (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-[(l,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)-5-[l(ethylcarbamoyl)azetidin-3-yl]-l,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 IM aqueous 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 13g RediSep column) eluting with a gradient of 5 - 95% acetonitrile in water afforded the desired product as a yellow solid (27.4 mg, 0.05 mmol, 66%) [as a lithium sait].
HRMS-ESI (m/z) [M+H]+ calcd for C23H25N8O3S2: 525.1486, found 525.1516.
Example 161: 2-({6-[(l,3-Benzothiazol-2-yl)amino]-5-methylpyridazin-3yl}(methyl)amino)-5-[(2S)-3-{4-[3-(dimethylamino)prop-l-yn-l-yI]-2-fluorophenoxy}-2methylpropyl] -l,3-thiazole-4-carboxylic acid
424
Step A: ethyl5-[(2S)-3-{4-[3-(dimethylaniino)prop-l-yn-l-yl]-2-fluorophenoxy}-2methylpropyl]-2-[methyl(5-methyl-6-{[(2Z)-3-{[2-(tritnethylsilyl)ethoxy]methyl}-2,3dihydro-l,3-benzothiazol-2-ylidene]amino}pyridazin-3-yl)amino]-l,3-thiazole-4-carboxylate
To a solution of the product from Préparation 3zf (40 mg, 0.09 mmol, 1 eq) in 1,4-dioxane (3 mL) was added Préparation 4a (37.6 mg, 0.09 mmol, 1 eq), A,A-diisopropylethylamine (0.05 mL, 0.28 mmol, 3 eq), césium carbonate (90.2 mg, 0.28 mmol, 3 eq) and Xantphos (10.7 mg, 0.02 mmol, 0.2 eq), followed by tris(dibenzylideneacetone)dîpalladium(0) (8.45 mg, 0.01 mmol, 0.1 eq) and the mixture was heated at 100 °C for 24 h then allowed to cool to ambient température. The reaction was partitioned between dichloromethane and water, and the organic phase was washed with brine, dried (magnésium sulfate), and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 4 g RediSep™ silica cartridge) eluting with a gradient of 0 - 7% methanol in dichloromethane afforded the desired product as a brown glass (30.7 mg, 0.04 mmol, 41%).
LC/MS (C4oH5oFN704SiS2) 804 [M+H]+; RT 1.59 (LCMS-V-B1) XH NMR (400 MHz, DMSO-d6) 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, 3H), 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-[(l,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)5-[(2S)-3-{4-[3-(dimethylamino)prop-l -yn-1 -yl]-2-fluorophenoxy}-2-methylpropyl]-l ,3thiazole-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 (IM 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 heated at reflux overnight. The reaction was partitioned between ethyl acetate and water, the aqueous phase was extracted with ethyl acetate (x2) then dichloromethane (x2), and the combined organic extracts were dried (magnésium sulfate) and concentrated in vacuo to afford the desired product that was used directly in the next step without further purification.
Step Ç: 2-({6-[(l,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)-5[(2S)-3-{4-[3-(dimethylamino)prop-l-yn-l-yl]-2-fluorophenoxy}-2-methylpropyl]-l,3thiazole-4-carboxylic acid
425
To a solution of the product from Step B (38 mg, 0.06 mmol, 1 eq) in 1,4-dioxane (3 mL) was added IM aqueous lithium hydroxide (0.56 mL, 0.56 mmol, 10 eq) and the mixture was heated at 100 °C for 2.5 h. The reaction was allowed to cool to ambient température and purification by reverse phase automated flash chromatography (CombiFlash Rf, C18 13g RediSep column) eluting with a gradient of 5 — 95% acetonitrile in water afforded a brown glass. The material 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 followed by methanol, then elution with 10% formic acid in dichoromethane, solvent removal, and drying under vacuum afforded the desired product as a brown glass (6.4 mg, 0.01 mmol, 18%) [as a formic acid sait].
HRMS-ESI (m/z) [M+H]+ calcd for C32H33FN7O3S2: 646.2065, found 646.2110.
Example 162: 2-({6-[(l,3-Benzothiazol-2-yl)amino]-5-methylpyridazin-3yl}(methyl)amino)-5-[(LE)-2-methyl-3-phenoxyprop-l-en-l-yI]-l,3-thiazole-4-carboxylic acid
Step A: ethyl 2-[methyl(5-methyl-6-{[(2Z)-3-{[2-(trimethylsilyl)ethoxy]methyl}-2,3-dihydrol,3-benzothiazol-2-ylidene]amino}pyridazin-3-yl)amino]-5-[(lE)-2-methyl-3-phenoxyprop1 -en-1 -yl]-l,3-thiazole-4-carboxylate
To a solution of the product from Préparation 5m (160 mg, 0.26 mmol, 1 eq) in toluene (3 mL) was added phénol (34 pL, 0.38 mmol, 1.5 eq), triphenylphosphine (100 mg, 0.38 mmol, 1.5 eq) and diisopropylazodicarboxylate (75 pL, 0.38 mmol, 1.5 eq) and the mixture was heated at 120 °C for 1 h under microwave irradiation. The reaction 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 cartridge) eluting with a gradient of 0 - 60% ethyl acetate in zso-heptane afforded the desired product as a yellow gum (100 mg, 0.14 mmol, 56%).
426 φ LC/MS (C35H42N6O4S1S2) 703 [Μ+Η]+; RT 3.12 (LCMS-V-C)
Ή NMR (400 MHz, DMSO-d6) 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 5 0.88 (m, 2H), -0.11 (s, 9H).
Step B: ethyl 2-({6-[(l,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)5-[(lE)-2-methyl-3-phenoxyprop-l-en-l-yl]-l,3-thiazole-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 uL, 2.12 mmol, 3 eq) and tetrabutylammonium fluoride 10 (IM in tetrahydrofuran; 2.12 mL, 2.12 mmol, 3 eq) and the mixture was heated at 70 °C overnight. The reaction was partitioned between ethyl acetate and water, and the organic phase was dried (magnésium sulfate) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 24 g RediSep™ silica cartridge) eluting with a gradient of 0 - 100% ethyl acetate in zso-heptane afforded the desired product as a yellow solid 15 (37 mg, 0.06 mmol, 9%).
LC/MS (C29H28N6O3S2) 573 [M+H]+; RT 2.65 (LCMS-V-C) ‘H NMR (400 MHz, DMSO-d6) δ 7.91 (br s, 1H), 7.73 (s, 1H), 7.43 - 7.35 (m, 2H), 7.34 - 7.28 (m, 3H), 7.25 - 7.16 (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-[(l,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)-5[(lE)-2-methyl-3-phenoxyprop-l-en-1 -yl]-l,3-thiazole-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 at 110 °C overnight. The reaction was allowed to cool to ambient température and purification 25 by automated flash column chromatography (CombiFlash Rf, 4 g RediSep™ silica cartridge) eluting with a gradient of 0 - 10% 7N methanolic ammonia in dichloromethane afforded a dark yellow solid. The solid was triturated with acetonitrile, filtered, and dried under vacuum to afford the desired product as a yellow solid (12.1 mg, 0.02 mmol, 34%).
HRMS-ESI (m/z) [M+H]+ calcd for C27H25N6O3S2: 545.1424, found 545.1426.
427
Example 163: 2-({6-[(l,3-Benzothiazol-2-yl)amino]-5-methylpyridazin-3yl}(methyl)amino)-5-[(LE')-3-{4-[3-(dimethylamino)prop-l-yn-l-yl]-2-fluorophenoxy}-2methylprop-l-en-l-yl]-l,3-thiazole-4-carboxylic acid
Step A: ethyl5-[(lE)-3-{4-[3-(dimethylamino)prop-l-yn-l-yl]-2-fluorophenoxy}-2methylprop-l-en-l-yl]-2-[methyl(5-methyl-6-{[(2Z)-3-{[2-(trimethylsilyl)ethoxy]methyl}-2,3dihydro-l,3-benzothiazol-2-ylidene]amino}pyridazin-3-yl)amino]-l,3-thiazole-4-carboxylate
To a solution of the product from Préparation 5m (618 mg, 0.99 mmol, 1 eq) in toluene (18 mL) was added the product from Préparation 6b (286 mg, 1.48 mmol, 1.5 eq), 10 triphenylphosphine (388 mg, 1.48 mmol, 1.5 eq) and diisopropylazodicarboxylate (291 pL, 1.48 mmol, 1.5 eq) and the mixture was heated at 120 °C for 1 h under microwave irradiation. The reaction 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 cartridge) 15 eluting with a gradient of 0 - 100% ethyl acetate in zso-heptane afforded the desired product as a brown oil (406 mg, 0.51 mmol, 51%).
LC/MS (C4oH48FN704SiS2) 802 [M+H]+; RT 2.68 (LCMS-V-C) *H NMR (400 MHz, DMSO-d6) 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, 20 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-[(l,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)5-[(lE)-3-{4-[3-(dimethylamino)prop-l-yn-l-yl]-2-fluorophenoxy}-2-methylprop-l-en-l-yl]l,3-thiazole-4-carboxylate
To a solution of the product from Step A (406 mg, 0.51 mmol, 1 eq) in acetonitrile (5 mL) was added pyridinium poly(hydrogen fluoride) (0.88 mL, 10.1 mmol, 20 eq) and the mixture was
428 heated at 60 °C overnight. The reaction was allowed to cool to ambient température then partxtioned between dichloromethane and 2N aqueous sodium hydroxide (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 cartridge) eluting with a gradient of 0 - 5% methanol in dichloromethane afforded the desired product as a yellow solid (139 mg, 0.21 mmol, 41%).
LC/MS (C34H34FN7O3S2) 672 [M+H]+; RT 2.07 (LCMS-V-C)
Ή NMR (400 MHz, DMSO-d6) 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-[(l,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)-5[(lE)-3-{4-[3-(dimethylamino)prop-l-yn-l-yl]-2-fluorophenoxy}-2-methylprop-l-en-l-yl]1,3-thiazole-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 at 110 °C for 4 h. The reaction was allowed to cool to ambient température and the solvent removed by rotary évaporation. Purification by automated flash column chromatography (CombiFlash Rf, 12 g RediSep™ silica cartridge) eluting with a gradient of 0 - 25% 7N methanolic ammonia in dichloromethane afforded a solid. The material was triturated with acetonitrile, filtered, and dried under vacuum to afford the desired product as a yellow solid (77.9 mg, 0.12 mmol, 59%).
HRMS-ESI (m/z) [M+H]+ calcd for C32H31FN7O3S2: 644.1908, found 644.1908.
Example 164: 2-({6-[(l,3-Benzothiazol-2-yl)amino]-5-methyIpyridazin-3yl}(methyl)amino)-5-(3-{4-[3-(dimethylamino)prop-l-yn-l-yl]-2-fluoropheiioxy}-2hydroxypropyl)-l,3-thiazole-4-carboxylic acid
429
\
Step A: ethyl 5-(3-{4-[3-(dimethylamino)prop-l-yn-1 -yl]-2-fluorophenoxy}-2-(oxan-2yloxy)propyl)-2-[methyl(5-methyl-6-{[(2Z)-3-{[2-(trimethylsilyl)ethoxy]methyl}-2,3-dihydrol,3-benzothiazol-2-ylidene]amino}pyridazin-3-yl)amino]-l,3-thiazole-4-carboxylate
The product from Préparation 4a (150 mg, 0.37 mmol, 1.16 eq) and the product from Préparation 3zg (165 mg, 0.32 mmol, 1 eq) were added to a solution of tris(dibenzylïdeneacetone)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). Césium carbonate (310 mg, 0.95 mmol, 3 eq) and Aj/V-diisopropylethylamine (0.2 mL, 0.95 mmol, 3 eq) were added and the mixture was 10 heated in a sealed tube at 125 °C for 18 h. The suspension was allowed to cool and the mixture 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) eluting with a gradient of 0 - 2.5% methanol in dichloromethane afforded the desired product as a dark brown gum (230 mg, 0.26 mmol, 81%).
LC/MS (C44H56FN7O6SiS2) 890 [M+H]+; RT 1.59 (LCMS-V-B1)
Step B: ethyl 2-({6-[(l,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)5-(3-{4-[3-(dimethylamino)prop-l -yn-1 -yl]-2-fluorophenoxy}-2-hydroxypropyl) -1,3thiazole-4-carboxylate
To a solution of the product from Step A (225 mg, 0.25 mmol, 1 eq) in chloroform (4.5 mL), 20 cooled in ice-water, was added trifluoroacetic acid (0.5 mL, 6.32 mmol, 25 eq) and the mixture was stirred for 1 h at 0 °C and for 18 h at ambient température. Dichloromethane (50 mL) was added and the mixture was washed successively with saturated aqueous sodium bicarbonate (30 mL), water (2 x 30 mL) and brine (30 mL), dried (magnésium sulfate), and concentrated in vacuo. Purification by flash column chromatography (20 g silica) eluting with a gradient of 0
430
5% methanol in dichloromethane afforded the desired product as a dark yellow solid (98 mg, 0.15 mmol, 57%).
LC/MS (C33H34FN7O4S2) 676 [M+H]+; RT 1.15 (LCMS-V-B1) JH NMR (400 MHz, DMSO-d6) δ 7.86 - 7.63 (m, 2H), 7.48 - 7.31 (m, 2H), 7.30 - 7.21 (m, 5 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 = 7.1Hz, 3H).
Step C: 2-({6-[(l,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)-5-(3{4-[3-(dimethylamino)prop-l -yn-1 -yl]-2-fluorophenoxy}-2-hydroxypropyl) -l,3-thiazole-410 carboxylic acid
To a solution of the product from Step B (90 mg, 0.13 mmol, 1 eq) in a 1:3 mix of 1,4-dioxane / water (2 mL) was added lithium hydroxide monohydrate (11.2 mg, 0.27 mmol, 2 eq) and the suspension was heated at 40 °C for 7 h. The reaction was allowed to cool and the resulting solution acidified with acetic acid to pH 5 to give a yellow precipitate. The solids were collected 15 by filtration, washed with water and dried under vacuum to afford the desired product as a yellow solid (54 mg, 0.08 mmol, 63%).
HRMS-ESI (m/z) [M+H]+ calcd for C31H31FN7O4S2: 648.1857, found 648.1888.
Example 165:2-[[6-(l,3-BenzothiazoI-2-ylamino)-5-methyI-pyridazm-3-yl]-[5-[methyl(4piperidyl)amino]pentyl]amino]-5-[3-[4-[3-(dimethylammo)prop-l-ynyl]-2-fluoro20 phenoxy]propyl]thiazole-4-carboxylic acid
Step A: methyl 2-[5-[(1 -tert-butoxycarbonyl-4-piperidyl)-methyl-amino]pentyl-[5-methyl-6[(Z)-[3-(2-trimethylsilylethoxymethyl)-l,3-benzothiazol-2-ylidene]amino]pyridazin-3
431 yl]amino]-5-[3-[4-[3-(dimethylamino)prop-l-ynyl]-2-fluoro-phenoxy]propyl]thiazole-4carboxylate
Using Alkylation with in situ generated tosylate General Procedure starting from
Préparation 5f and tert-butyl 4-(methylamino)piperidine-l-carboxylate as the appropriate 5 amine, the desired product was obtained.
HRMS-ESI (m/z): [M+2H]2+ calcd for C53H74FN9O6S2Si: 522.7551, found 522.7542.
Step B: 2-[[6-(l,3-benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-[5-[methyl(4piperidyl)amino]pentyl]amino]-5-[3-[4-[3-(dimethylamino)prop-l-ynyl]-2-fluorophenoxy]propyl]thiazole-4-carboxylic acid
Using Deprotection and Hydrolysis General Procedure followed by repurification via reverse phase préparative chromatography (C18, 0.1% TFA in water : MeCN) starting from the product from Step A, the TFA-salt of the desired product was obtained.
HRMS-ESI (m/z): [M+2H]2+ calcd for C4iH52FN9O3S2: 400.6804, found 400.6803.
Example 166:2-[[6-(l,3-Benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-[5-(215 morpholinoethylamino)pentyl]amino]-5-[3-[4-[3-(dimethylamino)prop-l-ynyI]-2-fluorophenoxy] propyl] thiazole-4-carboxy lie acid
Step A: methyl5-[3-[4-[3-(dimethylamino)prop-l-ynyl]-2-fluoro-phenoxy]propyl]-2-[[5methyl-6-[ (Z) -[3-(2-trimethylsilylethoxymethyl) -l,3-benzothiazol-220 ylidene]amino]pyridazin-3-yl]-[5-(2-morpholinoethylamino)pentyl]amino]thiazole-4carboxylate
Using Alkylation with in situ generated tosylate General Procedure starting from
432
B Préparation 5f and 2-morpholinoethanamine as the appropriate amine, the desired product was obtained.
JH NMR (500 MHz, DMSO-rfo) 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, 5 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); HRMS-ESI (m/z): [M+2H]2+ calcd for C48H68FN9O5S2Si: 480.7269, found 480.7264.
Step B: 2-[[6-(1,3-benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-[5-(210 morpholinoethylamino)pentyl]amino]-5-[3-[4-[3-(dimethylamino)prop-l-ynyl]-2-fluorophenoxy]propyl]thiazole-4-carboxylic acid
Using Deprotection and Hydrolysis General Procedure followed by repurification via reverse phase préparative chromatography (C18, 0.1% TFA in water : MeCN) starting from the product from Step A, the TFA-salt of the desired product was obtained.
HRMS-ESI (m/z): [M+2H]2+ calcd for C41H52FN9O4S2: 408.6779, found 408.6769.
Example 167:2-[[6-(l,3-Benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-[5-[2-(4methylpiperazin-l-yl)ethylamino]pentyl]amino]-5-[3-[4-[3-(dimethylamino)prop-lyny 1] -2-fluoro-phenoxy] propy 1] thiazole-4-carboxy lie acid
Step A: methyl 5-[3-[4-[3-(dimethylamino)prop-l-ynyl]-2-fluoro-phenoxy]propyl]-2-[5-[2(4-methylpiperazin-l-yl)ethylamino]pentyl-[5-methyl-6-[(Z)-[3-(2trimethylsilylethoxymethyl)-l,3-benzothiazol-2-ylidene]amino]pyridazin-320487
433
M yl]amino]thiazole-4-carboxylate
Using Alkylation with in situ generated tosylate General Procedure starting from Préparation 5f and 2-(4-methylpiperazin-l-yl)ethanamine as the appropriate amine, the desired product was obtained.
Ή NMR (500 MHz, DMSO-ife) 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-ESI (m/z):
[M+2H]2+ calcd for C49H7iFNio04S2Si: 487.2422, found 487.2422.
Step B: 2-[[6-(l,3-benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-[5-[2-(4methylpiperazin-1 -yl) ethylamino]pentyl]amino]-5-[3-[4-[3-(dimethylamino)prop -ynyl]-2fluoro-phenoxy]propyl]thiazole-4-carboxylic acid
Using Deprotection and Hydrolysis General Procedure followed by repurification via 15 reverse phase préparative chromatography (C18, 0.1% TFA in water : MeCN) starting from the product from Step A, the TFA-salt of the desired product was obtained.
HRMS-ESI (m/z): [M+2H]2+ calcd for C42H55FN10O3S2: 415.19366, found 415.1939.
Example 168:2-[[6-(l,3-Benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-[4-[2-(4methylpiperazin-l-yl)ethylamino]butyl]amino]-5-[3-[4-[3-(dimethylamino)prop-l-ynyl]20 2-fluoro-phenoxy]propyl]thiazole-4-carboxylic acid
434
Step A: methyl5-[3-[4-[3-(dimethylamino)prop-l-ynyl]-2-fluoro-phenoxy]propyl]-2-[4-[2(4-methylpiperazin-l-yl)ethylamino]butyl-[5-methyl-6-[(Z)-[3-(2trimethylsilylethoxymethyl)-l,3-benzothiazol-2-ylidene]amino]pyridazin-3yl]amino]thiazole-4-carboxylate
Using Alkylation with in situ generated tosylate General Procedure starting from Préparation 5d and 2-(4-methylpiperazin-l-yl)ethanamine as the appropriate amine, the desired product was obtained.
Ή NMR (500 MHz, DMSO-îZ6) 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); HRMS-ESI (m/z): [M+2H]2+ calcd for C48H69FN10O4S2S1: 480.2344, found 480.2340.
Step B: 2-[[6-(l,3-benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-[4-[2-(4methylpiperazin-l-yl)ethylamino]butyl]amino]-5-[3-[4-[3-(dimethylamino)prop-l-ynyl]-2fluoro-phenoxy]propyl]thiazole-4-carboxylic acid
Using Deprotection and Hydrolysis General Procedure followed by repurification via reverse phase préparative chromatography (C18, 0.1% TFA in water : MeCN) starting from the product from Step A, the TFA-salt of the desired product was obtained.
HRMS-ESI (m/z): [M+2H]2+ calculated for C41H53FN10O3S2: 408.1858, found 408.1857.
Example 169:2-[[6-(l,3-Benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-[4-hydroxy-5[methyl-[2-(methylamino)ethyl]amino]pentyl]amino]-5-[3-[4-[3-(dimethylamino)prop-lynyl]-2-fluoro-phenoxy]propyl]thiazole-4-carboxylic acid
435
Step A: methyl2-[[5-[2-[tert-butoxycarbonyl(methyl)amino]ethyl-methyl-amino]-4-[tertbutyl(diphenyl)silyl]oxy-pentyl]-[5-methyl-6-[(Z)-[3-(2-trimethylsilylethoxymethyl)-l,3benzothiazol-2-ylidene]amino]pyridazin-3-yl]amino]-5-[3-[4-[3-(dimethylamino)prop-l5 ynyl]-2-fluoro-phenoxy]propyl]thiazole-4-carboxylate
Using Alkylation with tosylate General Procedure starting from Préparation 5b and tertbutyl N-methyl-N-[2-(methylamino)ethylJcarbamate as the appropriate amine, the desired product was obtained.
Step^:2-[[6-(l,3-benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-[4-hydroxy-5-[methyl10 [2-(methylamino)ethyl]amino]pentyl]amino]-5-[3-[4-[3-(dimethylamino)prop-l-ynyl]-2fluoro-phenoxy]propyl]thiazole-4-carboxylic acid
Using Deprotection and Hydrolysis General Procedure starting from the product from Step A as the appropriate methyl ester, the desired product was obtained.
HRMS-ESI (m/z): [M+H]+ calcd for C39H49FN9O4S2: 790.3327, found 790.3317.
Example 170:2-iï6-(1.3-Benzothiazol-2-.ylamino)-5-methyl-pyridazin-3-yl]-[5-[2(diethylamino)ethylamino]-4-hydroxy-pentyl]amino]-5-[3-[4-[3-(dimethylamino)prop-lynyl]-2-fluoro-phenoxy]propyl]thiazole-4-carboxylic acid
436
Step A: methyl 2-[[4-[tert-butyl(diphenyl)silyl]oxy-5-[2-(diethylamino)ethylamino]pentyl][5-methyl-6-[(Z)-[3-(2-trimethylsilylethoxymethyl)-l,3-benzothiazol-2ylidene]amino]pyridazin-3-yl]amino]-5-[3-[4-[3-(dimethylamino)prop-l-ynyl]-2-fluoro5 phenoxy]propyl]thiazole-4-carboxylate
Using Alkylation with tosylate General Procedure starting from Préparation 5b and N',N'diethylethane-l,2-diamine as the appropriate amine, the desired product was obtained.
Step B: 2-[[6-(l,3-benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-[5-[2(diethylamino)ethylamino]-4-hydroxy-pentyl]amino]-5-[3-[4-[3-(dimethylamino)prop-l10 ynyl]-2-fluoro-phenoxy]propyl]thiazole-4-carboxylic acid
Using Deprotection and Hydrolysis General Procedure starting from the product from Step A as the appropriate methyl ester, the desired product was obtained.
HRMS-ESI (m/z): [M+H]+ calcd for C41H53FN9O4S2: 818.3640, found 818.3613.
Example 171:2-lï5-(4-Amino-l-piperidyl)-4-hydroxy-pentyl]-[6-(l,3-benzothiazol-215 ylamino)-5-methyl-pyridazin-3-yl]amino]-5-[3-[4-[3-(dimethylamino)prop-l-ynyl]-2fluoro-phenoxy] propy 1] thiazole-4-carboxylic acid
437
B Step A: methyl2-[[5-[4-(tert-butoxycarbonylamino)-1 -piperidyl]-4-[tertbutyl(diphenyl)silyl]oxy-pentyl]-[5-methyl-6-[(Z)-[3-(2-trimethylsilylethoxymethyl)-l,3benzothiazol-2-ylidene]amino]pyridazin-3-yl]amino]-5-[3-[4-[3-(dimethylamino)prop-lynyl]-2-fluoro-phenoxy]propyl]thiazole-4-carboxylate
Using Alkylation with tosylate General Procedure starting from Préparation 5b and tertbutyl N-(4-piperidyl)carbamate as the appropriate amine, the desired product was obtained.
Step B: 2-[[5-(4-amino-l-piperidyl)-4-hydroxy-pentyl]-[6-(l,3-benzothiazol-2-ylamino)-5methyl-pyridazin-3-yl]amino]-5-[3-[4-[3-(dimethylamino)prop-l-ynyl]-2-fluorophenoxy]propyl]thiazole-4-carboxylic acid
Using Deprotection and Hydrolysis General Procedure starting from the product from Step A as the appropriate methyl ester, the desired product was obtained.
HRMS-ESI (m/z): [M+H]+ calcd for C40H49FN9O4S2: 802.3327, found 802.3324.
Example 172:2-[[6-(l,3-Benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-(4-hydroxy-5piperazin-l-yl-pentyl)amino]-5-[3-[4-[3-(dimethylamino)prop-l-ynyl]-2-fluoro15 phenoxy]propyl]thiazole-4-carboxylic acid
Step A: methyl 2-[[5-(4-tert-butoxycarbonylpiperazin-l-yl)-4-[tert-butyl(diphenyl)silyl]oxypentyl]-[5-methyl-6-[(Z)-[3-(2-trimethylsilylethoxymethyl)-l,3-benzothiazol-2ylidene]amino]pyridazin-3-yl]amino]-5-[3-[4-[3-(dimethylamino)prop-l-ynyl]-2-fluoro20 phenoxy]propyl]thiazole-4-carboxylate
Using Alkylation with tosylate General Procedure starting from Préparation 5b and tertbutylpiperazine-l-carboxylate as the appropriate amine, the desired product was obtained.
Step B: 2-[[6-(l,3-benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-(4-hydroxy-520487
438
I piperazin-1 -yl-pentyl)amino]-5-[3-[4-[3-(dimethylamino)prop-l -ynyl]-2-fluorophenoxy]propyl]thiazole-4-carboxylic acid
Using Deprotection and Hydrolysis General Procedure starting from the product from Step A as the appropriate methyl ester, the desired product was obtained.
HRMS-ESI (m/z): [M+2H]2+ calcd for C39H48FN9O4S2: 394.6622, found 394.6614.
Example 173:2-[[6-(l,3-Benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-[4-hydroxy-5[2-(l-piperidyl)ethylamino]pentyl]amino]-5-[3-[4-[3-(dimethylamino)prop-l-ynyl]-2fluoro-phenoxy] propyl] thiazole-4-carboxyiic acid
Step A: methyl2-[[4-[tert-butyl(diphenyl)silyl]oxy-5-[2-(l-piperidyl)ethylamino]pentyl]-[5methyl-6-[(Z)-[3-(2-trimethylsilylethoxymethyl)-l,3-benzothiazol-2ylidene]amino]pyridazin-3-yl]amino]-5-[3-[4-[3-(dimethylamino)prop-l-ynyl]-2-fluoro· phenoxy]propyl]thiazole-4-carboxylate
Using Alkylation with tosylate General Procedure starting from Préparation 5b and 2-(l- piperidyl)ethanamine as the appropriate amine, the desired product was obtained.
Step B: 2-[[6-(l,3-benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-[4-hydroxy-5-[2-(lpiperidyl)ethylamino]pentyl]amino]-5-[3-[4-[3-(dimethylamino)prop-l-ynyl]-2-fluorophenoxy] propyl]thiazole-4-carboxylic acid
Using Deprotection and Hydrolysis General Procedure starting from the product from Step 20 A as the appropriate methyl ester, the desired product was obtained.
HRMS-ESI (m/z): [M+2H]2+ calcd for C42H54FN9O4S2: 415.6857, found 415.6853.
439
Example 174: 2-({6-[(l,3-Benzothiazol-2-yl)amino]-5-methylpyridazin-3yl}(methyl)ammo)-5-[(lE')-3-{2-fluoro-4-[3-(methylamino)prop-l-yn-l-yl]phenoxy}-2methylprop-l-en-l-yl]-l,3-thiazole-4-carboxylic acid
Step A: ethyl5-[(lE)-3-[4-(3-{[(tert-butoxy)carbonyl](methyl)amino}prop-l-yn-1 -yl)-2fluorophenoxy]-2-methylprop-l-en-l-yl]-2-[methyl(5-methyl-6-{[(2Z)-3-{[2(trimethylsilyl)ethoxy]methyl}-2,3-dihydro-l,3-benzothiazol-2-ylidene]amino}pyridazin-3yl)amino]-l,3-thiazole-4-carboxylate
To a solution of the product from Préparation 5m (641 mg, 1.02 mmol, 1 eq) in toluene (18 mL)was added the product from Préparation 6a (428 mg, 1.53 mmol, 1.5 eq), triphenylphosphine (402 mg, 1.53 mmol, 1.5 eq) and diisopropylazodicarboxylate (0.3 mL, 1.53 mmol, 1.5 eq) and the mixture was heated at 120 °C for 1 h. The reaction 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 cartridge) eluting with a gradient of 0 - 40% ethyl acetate in zso-heptane afforded the desired product as a yellow solid (380 mg, 0.43 mmol, 42%).
LC/MS (C44H54FN7O6SiS2) 888 [M+H]+; RT 1.43 (LCMS-V-B2)
Ή NMR (400 MHz, DMSO-d6) 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-[(l,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)5-[(l E) -3-{2-fluoro-4-[3-(methylamino)prop-l -yn-1 -yl]phenoxy}-2-methylprop-l -en-1 -yl]l,3-thiazole-4-carboxylate
440
P To a solution of the product from Step A (380 mg, 0.43 mmol, 1 eq) in acetonitrile (5 mL) was added pyridinium poly(hydrogen fluoride) (744 pL, 8.56 mmol, 20 eq) and the mixture was heated at 60 °C for 3 h. The reaction was allowed to cool to ambient température then partitioned between dichloromethane and 2N aqueous sodium hydroxide, the aqueous phase extracted with 5 1:3 isopropanol / dichloromethane, and the combined organics dried (PTFE phase separator) and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 12 g RediSep™ silica cartridge) eluting with a gradient of 0 - 6% methanol in dichloromethane afforded the desired product as a yellow solid (133 mg, 0.2 mmol, 47%).
LC/MS (C33H32FN7O3S2) 658 [M+H]+; RT 2.02 (LCMS-V-C) JH NMR (400 MHz, DMSO-d6) 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-[(l,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)-515 [(lE)-3-{2-fluoro-4-[3-(methylamino)prop-l-yn-l-yl]phenoxy}-2-methylprop-l-en-l-yl]-l,3thiazole-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 at 110 °C for 4 h. The reaction was allowed to cool to ambient température and purification by 20 automated flash column chromatography (CombiFlash Rf, 12 g RediSep™ silica cartridge) eluting with a gradient of 0 — 25% 7N methanolic ammonia in dichloromethane afforded a solid. The solid was triturated with acetonitrile, filtered, and dried under vacuum to afford the desired product as a yellow solid (4.32 mg, 0.01 mmol, 3%).
HRMS-ESI (m/z): [M+H]+ calcd for C31H29FN7O3S2: 630.1752, found 630.1781.
Example 175:2-[4-(4-Amino-l-piperidyl)butyl-[6-(l,3-benzothiazoI-2-ylamino)-5methyI-pyridazin-3-yl]amino]-5-[3-[4-[3-(dimethylamino)prop-l-ynyI]-2-fluorophenoxy]propyl]thiazole-4-carboxylic acid
441
Step A: methyl 2-[4-[4-(tert-butoxycarbonylamino)-l-piperidyl]butyl-[5-methyl-6-[(Z)-[3-(2trimethylsilylethoxymethyl)-l,3-benzothiazol-2-ylidene]amino]pyridazin-3-yl]amino]-5-[3[4-[3-(dimethylamino)prop-l-ynyl]-2-fluoro-phenoxy]propyl]thiazole-4-carboxylate
Using Alkylation with in situ generated tosylate General Procedure starting from Préparation 5d and tert-butyl N-(4-piperidyl)carbamate as the appropriate amine, the desired product was obtained.
Tl NMR (500 MHz, DMSO-î/6) 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), 10 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); HRMS-ESI (m/z): [M+H]+ calcd for C5iH7iFN9O6S2Si: 1016.4716, found 1016.4704.
Step B: 2-[4-(4-amino-l-piperidyl)butyl-[6-(l,3-benzothiazol-2-ylamino)-5-methylpyridazin-3-yl]amino]-5-[3-[4-[3-(dimethylamino)prop-l-ynyl]-2-fluoro15 phenoxy]propyl]thiazole-4-carboxylic acid
Using Deprotection and Hydrolysis General Procedure followed by repurification via reverse phase préparative chromatography (C18, 0.1% TFA in water : MeCN) starting from the product from Step A, the TFA-salt of the desired product was obtained.
HRMS-ESI (m/z): [M+H]+ calcd for C39H46FN9O3S2: 772.3222 found 772.3228.
Example 176:2- [[6- (l,3-Benzothiazol-2-y lamino) -5-methyl-pyridazin-3-y 1] - [4- [2(diethylamino)ethylamino]butyl]amino]-5-[3-[4-[3-(dimethylamino)prop-l-ynyl]-2fluoro-phenoxy]propyl]thiazole-4-carboxyIic acid
442
Step A: methyl 2-[4-[2-(diethylamino)ethylamino]butyl-[5-methyl-6-[(Z)-[3-(2trimethylsilylethoxymethyl)-l,3-benzothiazol-2-ylidene]amino]pyridazin-3-yl]amino]-5-[3[4-[3-(dimethylamino)prop-l-ynyl]-2-fluoro-phenoxy]propyl]thiazole-4-carboxylate
Using Alkylation with in situ generated tosylate General Procedure starting from Préparation 5d and N',N'-diethylethane-l,2-diamine as the appropriate amine, the desired product was obtained.
XH NMR (500 MHz, DMSO-î/6) 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, 10 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-ESI (m/z): [M+H]+ calcd for C47H67FN9O4S2S1: 932.4505 found 932.4489.
Step B: 2-[[6-(l,3-benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-[4-[215 (diethylamino)ethylamino]butyl]amino]-5-[3-[4-[3-(dimethylamino)prop-l-ynyl]-2-fluorophenoxy]propyl]thiazole-4-carboxylic acid
Using Deprotection and Hydrolysis General Procedure followed by repurification via reverse phase préparative chromatography (C18, 0.1% TFA in water : MeCN) starting from the product from Step A, the TFA-salt of the desired product was obtained.
HRMS-ESI (m/z): [M+2H]2+ calcd for C40H52FN9O3S2: 394.6804, found 394.6802.
Example 177:2-r5-(4-Amino-l-piperidyl)pentyl-[6-(l,3-benzothiazol-2-ylamino)-5methyl-pyridazin-3-yI]amino]-5-[3-[4-[3-(dimethylamino)prop-l-ynyl]-2-fluorophenoxy]propyl]thiazole-4-carboxylic acid
443
\
Step A: methyl2-[5-[4-(tert-butoxycarbonylamino)-l-piperidyl]pentyl-[5-methyl-6-[(Z)-[3(2-trimethylsilylethoxymethyl)-l,3-benzothiazol-2-ylidene]amino]pyridazin-3-yl]amino]-5[3-[4-[3-(dimethylamino)prop-l-ynyl]-2-fluoro-phenoxy]propyl]thiazole-4-carboxylate
Using Alkylation with in situ generated tosylate General Procedure starting from Préparation 5f and tert-butyl N-(4-piperidyl)carbamate as the appropriate amine, the desired product was obtained.
Ή NMR (500 MHz, DMSO-d6) 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 10 (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-ESI (m/z): [M+2H]2+ calcd for C52H74FN9O6S2Si: 515.7473, found 515.74725.
Step B: 2-[5-(4-amino-l-piperidyl)pentyl-[6-(l,3-benzothiazol-2-ylamino)-5-methyl15 pyridazin-3-yl]amino]-5-[3-[4-[3-(dimethylamino)prop-l-ynyl]-2-fluorophenoxy]propyl]thiazole-4-carboxylic acid
Using Deprotection and Hydrolysis General Procedure followed by repurification via reverse phase préparative chromatography (C18, 0.1% TFA in water : MeCN) starting from the product from Step A, the TFA-salt of the desired product was obtained.
HRMS-ESI (m/z): [M+2H]2+ calcd for C4oH50FN903S2: 393.6726, found 393.6723.
444
Example 178:2-[[6-(l,3-Benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-[4-[methyl-[2(methylamino)ethyl]amino]butyl]amino]-5-[3-[4-[3-(dimethylamino)prop-l-ynyl]-2fluoro-phenoxy]propyl]thiazole-4-carboxylic acid
Step A: methyl 2-[4-[2-[tert-butoxycarbonyl(methyl)amino]ethyl-methyl-amino]butyl-[5methyl-6-[(Z)-[3-(2-trimethylsilylethoxy methyl)-1,3-benzothiazol-2ylidene]amino]pyridazin-3-yl]amino]-5-[3-[4-[3-(dimethylamino)prop-l-ynyl]-2-fluorophenoxy]propyl]thiazole-4-carboxylate
Using Alkylation with in situ generated tosylate General Procedure starting from 10 Préparation 5d and tert-butyl N-methyl-N-[2-(methylamino)ethyl]carbamate as the appropriate amine, the desired product was obtained.
XH NMR (500 MHz, DMSO-d6) 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 15 (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); HRMS-ESI (m/z): [M+H]+ calcd for C50H71FN9O6S2SÊ 1004.4716, found 502.73931.
Step B: 2-[[6-(l,3-benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-[4-[methyl-[2(methylamino)ethyl]amino]butyl]amino]-5-[3-[4-[3-(dimethylamino)prop-l-ynyl]-2-fluoro20 phenoxy]propyl]thiazole-4-carboxylic acid
Using Deprotection and Hydrolysis General Procedure followed by repurification via reverse phase préparative chromatography (C18, 0.1% TFA in water : MeCN) starting from the product from Step A, the TFA-salt of the desired product was obtained.
HRMS-ESI (m/z): [M+2H]2+ calcd for C38H48FN9O3S2: 380.6647, found 380.6640.
445
Example 179:2-[[6-(l,3-Benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-[4-[2-(lpiperîdyl)ethylamino] butyl]amino] -5- [3- [4- [3-(dimethyIamino)prop-l-ynyl] -2-fluorophenoxy]propyl]thiazo!e-4-carboxylic acid
Step A: methyl 5-[3-[4-[3-(dimethylamino)prop-l-ynyl]-2-fluoro-phenoxy]propyl]-2-[[5methyl-6-[(Z)-[3-(2-trimethylsilylethoxymethyl)-l,3-benzothiazol-2ylidene]amino]pyridazin-3-yl]-[4-[2-(l-piperidyl)ethylamino]butyl]amino]thiazole-4carboxylate
Using Alkylation with in situ generated tosylate General Procedure starting from 10 Préparation 5d and 2-(l -piperidyl)ethanamine as the appropriate amine, the desired product was obtained.
*H NMR (500 MHz, DMSO-rfe) 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, 15 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); HRMS-ESI (m/z): [M+H]+ calcd for C48H67FN9O4S2Si: 944.4505, found 944.44978.
Step B: 2-[[6-(l,3-benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-[4-[2-(lpiperidyl)ethylamino]butyl]amino]-5-[3-[4-[3-(dimethylamino)prop-l-ynyl]-2-fluoro20 phenoxy]propyl]thiazole-4-carboxylic acid
Using Deprotection and Hydrolysis General Procedure followed by repurification via reverse phase préparative chromatography (C18, 0.1% TFA in water : MeCN) starting from the product from Step A, the TFA-salt of the desired product was obtained.
HRMS-ESI (m/z): [M+H]+ calcd for C4iH5iFN9O3S2: 800.3535, found 800.3540.
446
Example 180:2-[[6-(l,3-Benzothiazol-2-ylamino)-5-methyI-pyridazin-3-yl]-[4-(2morpholinoethylamino)butyI]amino]-5-[3-[4-[3-(dimethylamino)prop-l-ynyl]-2-fluorophenoxy]propyl]thiazole-4-carboxylic acid
Step A: methyl 5-[3-[4-[3-(dimethylamino)prop-l-ynyl]-2-fluoro-phenoxy]propyl]-2-[[5methyl-6-[(Z)-[3-(2-trimethylsilylethoxymethyl)-l,3-benzothiazol-2ylidene]amino]pyridazin-3-yl]-[4-(2-morpholinoethylamino)butyl]amino]thiazole-4carboxylate
Using Alkylation with in situ generated tosylate General Procedure starting from 10 Préparation 5d and 2-morpholinoethanamine as the appropriate amine, the desired product was obtained.
fil NMR (500 MHz, DMSO-ife) 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, 3H), 2.39 (brm, 4H), 2.13 (m, 2H), 1.84-1.68 (m, 4H), 0.92 (m, 2H), -0.11 (s, 9H); HRMS-ESI (m/z): [M+2H]2+ calcd for C47H66FN9O5S2Si: 473.7186, found 473.7183.
Step B: -[[6-(l,3-benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-[4-(2morpholinoethylamino)butyl]amino]-5-[3-[4-[3-(dimethylamino)prop-l-ynyl]-2-fluoro20 phenoxy]propyl]thiazole-4-carboxylic acid
Using Deprotection and Hydrolysis General Procedure followed by repurification via reverse phase préparative chromatography (C18, 0.1% TFA in water : MeCN) starting from the product from Step A, the TFA-salt of the desired product was obtained.
HRMS-ESI (m/z): [M+2H]2+ calcd for C40H50FN9O4S2: 401.6700, found 401.6697.
447
Example 181:2-[[6-(l,3-Benzothiazol-2-yIamino)-5-methyl-pyridazin-3-yl]-[4-[(l-methyl4-piperidyl)amino] butyl]amino] -5 - [3 - [4- [3-(dimethylamino)prop-l-ynyl] -2-fluorophenoxy] propyl] thiazole-4-carboxylic acid
Step A: methyl 5-[3-[4-[3-(dimethylamino)prop-l-ynyl]-2-fluoro-phenoxy]propyl]-2-[4-[(lmethyl-4-piperidyl)amino]butyl-[5-methyl-6-[(Z)-[3-(2-trimethylsilylethoxymethyl)-l,3benzothiazol-2-ylidene]amino]pyridazin-3-yl]amino]thiazole-4-carboxylate
Using Alkylation with in situ generated tosylate General Procedure starting from Préparation 5d and l-methylpiperidin-4-amine as the appropriate amine, the desired product 10 was obtained.
XH NMR (500 MHz, DMSO-ifc) 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 15 (m+m, 4H), 1.46 (m, 2H), 0.92 (m, 2H), -0.11 (s, 9H); HRMS-ESI (m/z): [M+2H]2+ calcd for
C47H64FN9O4S2Si: 465.7211, found 465.7210.
Step B: 2-[[6-(l,3-benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-[4-[(l-methyl-4piperidyl)amino]butyl]amino]-5-[3-[4-[3-(dimethylamino)prop-l-ynyl]-2-fluorophenoxy]propyl]thiazole-4-carboxylic acid
Using Deprotection and Hydrolysis General Procedure followed by repurification via reverse phase préparative chromatography (C18, 0.1% TFA in water : MeCN) starting from the product from Step A, the TFA-salt of the desired product was obtained.
HRMS-ESI (m/z): [M+2H]2+ calcd for C4oH48FN903S2: 393.6726, found 393.6735.
448
Example 182:2-[[6-(l,3-Benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-[5-[2(diethylamino)ethylamino]pentyl]amino]-5-[3-[4-[3-(dimethylamino)prop-l-ynyI]-2fluoro-phenoxy]propyl]thiazole-4-carboxylic acid
H
\
Step A: methyl 2-[5-[2-(diethylamino)ethylamino]pentyl-[5-methyl-6-[(Z)-[3-(2trimethylsilylethoxymethyl)-l,3-benzothiazol-2-ylidene]amino]pyridazin-3-yl]amino]-5-[3[4-[3-(dimethylamino)prop-l-ynyl]-2-fluoro-phenoxy]propyl]thiazole-4-carboxylate
Using Alkylation with in situ generated tosylate General Procedure starting from Préparation 5f and N(N'-diethylethane-l,2-diamine as the appropriate amine, the desired product was obtained.
HRMS-ESI (m/z): [M+2H]2+ calcd for C48H7oFN904S2Si: 473.7367, found 473.7360.
Step B: 2-[[6-(l,3-benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-[5-[2(diethylamino)ethylamino]pentyl]amino]-5-[3-[4-[3-(dimethylamino)prop-l-ynyl]-2-fluorophenoxy]propyl]thiazole-4-carboxylic acid
Using Deprotection and Hydrolysis General Procedure followed by repurification via reverse phase préparative chromatography (C18, 0.1% TFA in water : MeCN) starting from the product from Step A, the TFA-salt of the desired product was obtained.
HRMS-ESI (m/z): [M+H]+ calcd for C41H53FN9O3S2: 802.3691, found 802.3702.
Example 183: 2-({6-[(l,3-Benzothiazol-2-yl)amino]-5-methylpyridazin-3yl}(methyl)amino)-5-(3-{4-[3-(dimethylamino)prop-l-yn-l-yl]-2-fluorophenoxy}-2methoxypropyl)-l,3-thiazole-4-carboxylic acid
449
\
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-l,3-benzothiazol-2ylidene]amino}pyridazin-3-yl)amino]-l,3-thiazole-4-carboxylate
The product from Préparation 4a (1.6 g, 3.93 mmol, 1 eq) and the product from Préparation 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). Césium carbonate (3.84 g, 11.8 mmol, 3 eq) and A/TV-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 filtered through celite, the solids washed with 1,4-dioxane (30 mL) and the filtrate was concentrated in vacuo. Purification by flash column chromatography (100 g silica) eluting with a gradient of 0 - 25% ethyl acetate in Ao-heptane afforded the desired product as a pale yellow solid (2.49 g, 3.11 mmol, 79%).
LC/MS (C38H6oN605Si2S2) 801 [M+H]+; RT 1.74 (LCMS-V-B2)
Hl NMR (400 MHz, DMSO-d6) 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), 1.16 - 1.02 (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-l,3-benzothiazol-2-ylidene]amino}pyridazin-3yl)amino]-l,3-thiazole-4-carboxylate
450
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 (IM in tetrahydrofuran; 10 mL, 9.17 mmol, 3 eq) and the mixture was stirred for 60 min at 0 °C. The reaction was partitioned between water (75 mL) and ethyl acetate (150 mL), and the organic phase was successively 5 washed with water (2 x 75 mL) and brine (75 mL), dried (magnésium sulfate) and concentrated in vacuo. Purification by flash column chromatography (50 g silica) eluting with a gradient of 0 - 50% ethyl acetate in Ao-heptane afforded the desired product as a pale yellow/ green foam (1.83 g, 2.84 mmol, 93%).
LC/MS (C29H4oN605SiS2) 645 [M+H]+; RT 1.56 (LCMS-V-B1)
Ή NMR (400 MHz, DMSO-d6) 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-615 {[(2Z)-3-{[2-(trimethylsilyl)ethoxy]methyl}-2,3-dihydro-l,3-benzothiazol-2ylidene]amino}pyridazin-3-yl)amino]-l,3-thiazole-4-carboxylate
To a solution of the product from Step B (750 mg, 1.16 mmol, 1 eq) in toluene (15 mL) was added diisopropylazodicarboxylate (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 20 heated at 100 °C for 18 h then allowed to cool to ambient température and concentrated in vacuo. Purification by flash column chromatography (100 g silica) eluting with a gradient of 0 - 35% ethyl acetate in Ao-heptane afforded the desired product as an orange gum (1.2 g, 1.04 mmol, 90%).
LC/MS (C35H42FIN6O5SiS2) 865 [M+H]+; RT 1.78 (LCMS-V-B1)
Ή NMR (400 MHz, DMSO-d6) 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-l-yn-l-yl]-2-fluorophenoxy}-230 methoxypropyl) -2-[methyl(5-methyl-6-{[ (2Z) -3 ~{[2-(trimethylsilyl) ethoxy]methyl}-2,3dihydro-l,3-benzothiazol-2-ylidene]amino}pyridazin-3-yl)amino]-l,3-thiazole-4-carboxylate
451
To a solution of the product from Step C (1.2 g, 1.04 mmol, 1 eq) in dimethylformamide (15 mL) was added copper(I) 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(prop2-yn-l-yl)amine (0.25 mL, 2.32 mmol, 2.23 eq). MN-Diisopropylethylarnine (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 température and was partitioned between ethyl acetate (100 mL) and water (75 mL), and the organic phase was successively washed with water (2 x 75 mL) and brine (75 mL), dried (magnésium sulfate) and concentrated in vacuo. Purification by flash column chromatography (50 g silica) eluting with a gradient of 0 - 100% ethyl acetate in iso10 heptane afforded the desired product as a brown gum (460 mg, 0.56 mmol, 54%).
LC/MS (C4oH5oFN705SiS2) 820 [M+H]+; RT 1.52 (LCMS-V-B1) ‘H NMR (400 MHz, DMSO-d6) 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), 4.01 - 3.89 (m, 1H), 3.86 (s, 3H), 3.80 - 3.68 (m, 2H), 3.61 15 - 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-[(l,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)5-(3-{4-[3-(dimethylamino)prop-l-yn-l-yl]-2-fluorophenoxy}-2-methoxypropyl)-l,3thiazole-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 température. Dichloromethane (150 mL) was added and the mixture was washed successively with 10% aqueous potassium carbonate (75 mL), water (2 x 75 mL) and brine (75 mL), dried (magnésium sulfate) and concentrated in vacuo. Trituration with diethyl 25 ether (10 mL) gave a solid that was collected by filtration, washed with diethyl ether (2 x 10 mL) and dried under vacuum to afford the desired product as a green powder (250 mg, 0.33 mmol, 58%).
LC/MS (C34H36FN7O4S2) 690 [M+H]+; RT 1.23 (LCMS-V-B1)
Hl NMR (400 MHz, DMSO-d6) δ 7.83 - 7.69 (m, 2H), 7.62 - 7.53 (m, 1H), 7.51 - 7.39 (m, 30 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).
452
Step F: 2-({6-[(l,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)-5-(3{4-[3-(dimethylamino)prop-l-yn-l-yl]-2-fluorophenoxy}-2-methoxypropyl)-l,3-thiazole-4carboxylic acid
To a solution of the product from Step E (240 mg, 0.31 mmol, 1 eq) in a 1:2 mixture of 1,45 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 h. The reaction was allowed to cool and was filtered through celite and the solids washed with water (4 mL). The filtrate was acidified with acetic acid to pH 6 to give a suspension and the solids were collected by filtration, washed with water (20 mL) and dried under vacuum to afford the desired product as a green solid (110 mg, 0.17 10 mmol, 53%).
HRMS-ESI (m/z) [M+H]+ calcd for C32H33FN7O4S2: 662.2014, found 662.2052.
Example 184:2-[[6-(l,3-Benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yI]-(4-piperazinl-ylbutyl)amino] -5- [3- [4- [3-(dimethylamino)prop-l-ynyl] -2-fluorophenoxy] propyl] thiazole-4-carboxylie acid
Step A: methyl 2-[4-(4-tert-butoxycarbonylpiperazin-l-yl)butyl-[5-methyl-6-[(Z)-[3-(2trimethylsilylethoxymethyl)-l,3-benzothiazol-2-ylidene]amino]pyridazin-3-yl]amino]-5-[3[4-[3-(dimethylamino)prop-l-ynyl]-2-fluoro-phenoxy]propyl]thiazole-4-carboxylate
Using Alkylation with in situ generated tosylate General Procedure starting from 20 Préparation 5d and tert-butyl piperazine-l-carboxylate as the appropriate amine, the desired product was obtained.
HRMS-ESI (m/z): [M+H]+ calcd for C5oH69FN906S2Si: 1002.4560, found 1002.4544.
Step B: 2-[[6-(l,3-benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-(4-piperazin-l20487
453 ylbutyl)aniino]-5-[3-[4-[3-(dimethylamino)prop-l-ynyl]-2-fluoro-phenoxy]propyl]thiaz.ole4-carboxyhc acid
Using Deprotection and Hydrolysis General Procedure followed by repurification via reverse phase préparative chromatography (C18, 0.1% TFA in water : MeCN) starting from the product from Step A, the TFA-salt of the desired product was obtained.
HRMS-ESI (m/z): [M+2H]2+ calcd for C38H46FN9O3S2: 379.6569, found 379.6565.
Example 185:2-[[6-(l,3-Benzothiazol-2-yIamino)-5-methyl-pyridazin-3-yl]-(4hydroxybutyl)amino]-5-[3-[2-fluoro-4-[(E)-3-(methylamino)prop-lenyI] phenoxy] propy 1] thiazole-4-carboxylic acid
Step A: tert-butylN-methyl-N-[(E)-3-(4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2yl)allyl] carbamate
The mixture of 1.50 g of tert-butylN-methyl-N-prop-2-ynyl-carbamate (8.86 mmol, 1 eq.), 1.70 g of 4,4,5,5-tetramethyl-l,3,2-dioxaborolane (13.30 mmol, 1.5 eq.), 90 mg of N,Ndiethylethanamine (0.89 mmol, 0.1 eq.) and 229 mg of chloridobis(q5cyclopentadienyl)hydridozirconium (Schwartz’s reagent) (0.89 mmol, 0.1 eq.) were stirred at 65°C for overnight. After the reaction EtOAc and cc. NH4C1 were added the organic layer was separated and washed with cc. NaHCO3, cc. NaCl, dried over MgSÛ4, filtered then concentrated onto Celite. It was purified via flash column chromatography using heptane and EtOAc as eluents to give 1347 mg (51%) of the desired product as a colorless thick oil.
XH NMR (500 MHz, DMSO-î/6) 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); 13C NMR (125 MHz, DMSO-d6) δ ppm 155.3, 148.9, 119.1, 83.4, 79.1, 52.2, 34.5, 28.5, 25.1; HRMS-EI (m/z): [M-lBu+H]+ calcd for C11H20BNO4: 241.1480, found 241.1597.
454
Step B: methyl 5-[3-[4-[(E)-3-[tert-butoxycarbonyl(methyl)amino]prop-l-enyl]-2-fluorophenoxy]propyl]-2-[4-[tert-butyl(dimethyl)silyl]oxybutylamino]thiazole-4-carboxylate .00 g of Préparation 3c, Step B (1.61 mmol, 1 eq.), 716 mg of the product from Step A (2.41 mmol, 1.5 eq.), 93 mg of Pd(PPh3)4 (0.08 mmol, 0.05 eq.) and 666 mg of K2CO3 (4.82 mmol, 3 eq.) were mixed in 15 mL of 1,2-dimethoxyethane and 1.9 mL of water in a MW vessel and irradiated at 110°C for 1 h. Full conversion was observed. EtOAc and water were added the organic layer was separated and washed with cc. NaCl, dried over MgSO4, filtered then concentrated onto Celite. It was purified via flash column chromatography using heptane and EtOAc as eluents to give 708 mg (66%) of the desired product as a colorless thick oil.
Ή NMR (500 MHz, DMSO-d6) 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); 13C NMR (125 MHz, DMSO-dô) 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; HRMS-ESI (m/z): [M+H]+ calcd for C33H53FN3O6SS1: 666.3403, found 666.3409.
Step C: methyl 5-[3-[4-[(E)-3-[tert-butoxycarbonyl(methyl)amino]prop-l-enyl]-2-fluorophenoxy]propyl]-2-[4-[tert-butyl(dimethyl)silyl]oxybutyl-[5-methyl-6-[(Z) -[3-(2trimethylsilylethoxymethyl)-l,3-benzothiazol-2-ylidene]amino]pyridazin-3yl]amino]thiazole-4-carboxylate
Using Buchwald General Procedure II starting from 138 mg of the product from Step B (0.21 mmol, 1 eq.) and 101 mg of Préparation 4a (0.25 mmol, 1.2 eq.) as the appropriate halide, 102 mg (48%) of the desired product was obtained.
*H NMR (500 MHz, DMSO-ifc) 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); HRMS-ESI (m/z): [M+H]+ calcd for C51H75FN7O7S2S12: 1036.4686, found 1036.4706.
Step D: 2-[[6-(l,3-benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-(4hydroxybutyl)amino]-5-[3-[2-fluoro-4-[(E)-3-(methylamino)prop-lenyl]phenoxy]propyl]thiazole-4-carboxylic acid
455
Using Deprotection and Hydrolysis General Procedure followed by repurification via reverse phase préparative chromatography (C18, 0.1% TFA in water : MeCN) starting from the product from Step C, the TFA-salt of the desired product was obtained in the isomeric ratio of ca. 82% E and 18% Z (based on NMR).
HRMS-ESI (m/z): [M+H]+ calcd for C33H37FN7O4S2: 678.2327, found 678.2326.
Example 186:2-[[6-(l,3-Benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-(4hydroxybutyl)amino]-5-[3-[2-fluoro-4-[3-(methylamino)propyl]phenoxy]propyl]thiazole4-carboxylic acid
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 bottle equipped with a PTFE-coated magnetic stirring bar was charged with 400 mg of Example 185, Step B (0.60 mmol, 1 eq.), 64 mg of 10w% Pd/C (0.06 mmol, 0.1 eq.) in 6 mL of methanol, and then placed under a nitrogen atmosphère using hydrogénation System. After that it was filled with 4 bar H2 gas and stirred at rt for 3 h. Full conversion was observed, Celite was added to the reaction mixtures and the volatiles were removed under reduced pressure. Then it was purified via flash column chromatography using heptane and EtOAc as eluents to give 381 mg (95%) of the desired product as colorless oil.
Hl NMR (500 MHz, DMSO-·) 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); 13C NMR (125 MHz, DMSO-·) 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; HRMS-ESI (m/z): [M+H]+ calcd for
456
C33H55FN3O6SSK 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-[(Z)-[3-(2trimethylsilylethoxymethyl)-l,3-benzothiazol-2-ylidene]amino]pyridazin-3- yl]amino]thiazole-4-carboxylate
Using Buchwald General Procedure II starting from 375 mg of the product from Step A (0.56 mmol, 1 eq.) and 274 mg of Préparation 4a (0.67 mmol, 1.2 eq.) as the appropriate halide, 479 mg (82%) of the desired product was obtained.
ΧΗ NMR (500 MHz, DMSO-d6) δ ppm 7.80 (dm, 1H), 7.65/7.61 (s/s, 1H), 7.46 (dd, 1H), 7.43 10 (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); 13C NMR (125 MHz, DMSO4) δ ppm 155.4/155.3, 127.1, 124.6, 123.4, 123.1, 117.8, 116.2, 15 116.1,111.9, 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-ESI (m/z): [M+H]+ calcd for C5iH77FN7O7S2Si2: 1038.4843, found 1038.4837.
Step C: 2-[[6-(l,3-benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-(4hydroxybutyl)amino]-5-[3~[2-fluoro-4-[3-(methylamino)propyl]phenoxy]propyl]thiazole-420 carboxylic acid
Using Deprotection and Hydrolysis General Procedure followed by repurification via reverse phase préparative chromatography (C18, 0.1% TFA in water : MeCN) starting from the product from Step B, the TFA-salt of the desired product was obtained.
HRMS-ESI (m/z): [M+H]+ calcd for C33H39FN7O4S2: 680.2483, found 680.2469.
Example 187:2- ir6-(l,3-Benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl] - [5- [2-(lpiperidyl)ethylamino]pentyl]amino]-5-[3-[4-[3-(dimethylamino)prop-l-ynyl]-2-fluorophenoxy]propyl]thiazole-4-carboxylic acid
457
H
Step A: methyl 5-[3-[4-[3-(dimethylamino)prop-l-ynyl]-2-fluoro-phenoxy]propyl]-2-[[5methyl-6-[(Z)-[3-(2-trimethylsilylethoxy methyl)-l,3-benzothiazol-2ylidene]amino]pyridazin-3-yl]-[5-[2-(l-piperidyl)ethylamino]pentyl]amino]thiazole-4- carboxylate
Using Alkylation with in situ generated tosylate General Procedure starting from Préparation 5f and 2-(l-piperidyl)ethanamine as the appropriate amine, the desired product was obtained.
HRMS-ESI (m/z): [M+2H]2+ calcd for C49H7oFN904S2Si: 479.7367, found 479.7353.
Step B: 2-[[6-(l,3-benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-[5-[2-(lpiperidyl)ethylamino]pentyl]amino]-5-[3-[4-[3-(dimethylamino)prop-l-ynyl]-2-fluorophenoxy]propyl]thiazole-4-carboxylic acid
Using Deprotection and Hydrolysis General Procedure followed by repurification via reverse phase préparative chromatography (C18, 0.1% TFA in water : MeCN) starting from the 15 product from Step A, the TFA-salt of the desired product was obtained.
HRMS-ESI (m/z): [M+H]+ calcd for C42H53FN9O3S2: 814.3691, found 814.3670.
Example 188:2-[[6-(l,3-Benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-[4(methylamino)butyl]amino] -5- [3- [4- [3-(dimethylamino)prop-l-ynyl] -2-fluorophenoxy] propyl] thiazole-4-carboxylic acid
458
Step A: methyl5-[3-[4-[3-(dimethylamino)prop-l-ynyl]-2-fluoro-phenoxy]propyl]-2-[4(methylamino)butyl-[5-methyl-6-[(Z)-[3-(2-trimethylsilylethoxymethyl)-l,3-benzothiazol-2ylidene]amino]pyridazin-3-yl]amino]thiazole-4-carboxylate
Using Alkylation with in situ generated tosylate General Procedure starting from Préparation 5d and methanamine as the appropriate amine, the desired product was obtained.
Ή NMR (500 MHz, DMSO-î/6) δ ppm 7.84 (dm, 1H), 7.72 (s, 1H), 7.47 (dm, 1H), 7.44 (m, 1H), 7.31 (dd, 1 H), 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, 10 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); HRMSESI (m/z): [M+H]+ calcd for C42H56FN8O4S2Si: 847.3614, found 847.3594.
Step B: 2-[[6-(l,3-benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-[4(methylamino)butyl]amino]-5-[3-[4-[3-(dimethylamino)prop-l-ynyl]-2-fluorophenoxy]propyl]thiazole-4-carboxylic acid
Using Deprotection and Hydrolysis General Procedure followed by repurification via reverse phase préparative chromatography (C18, 0.1% TFA in water : MeCN) starting from the product from Step A, the TFA-salt of the desired product was obtained.
HRMS-ESI (m/z): [M+2H]2+ calcd for C35H4iFN8O3S2: 352.1358, found 352.1361.
Example 189:2-iï6-(L3-Benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-(5-piperazin20 l-ylpentyl)amino]-5-[3-[4-[3-(dimethylamino)prop-l-ynyl]-2-fluorophenoxy]propyl]thiazole-4-carboxylic acid
459
HN
Step A: methyl2-[5-(4-tert-butoxycarbonylpiperazin-l-yl)pentyl-[5-methyl-6-[(Z)-[3-(2trimethylsilylethoxymethyl)-l,3-benzothiazol-2-ylidene]amino]pyridazin-3-yl]amino]-5-[3[4-[3-(dimethylamino)prop-l-ynyl]-2-fluoro-phenoxy]propyl]thiazole-4-carboxylate
Using Alkylation with in situ generated tosylate General Procedure starting from Préparation 5f and tert-butyl piperazine-l-carboxylate as the appropriate amine, the desired product was obtained.
HRMS-ESI (m/z): [M+H]+ calcd for CsiHyiFNgOôSzSi: 1016.4716, found 1016.4710.
Step B: 2-[[6-(l,3-benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-(5-piperazin-l10 ylpentyl)amino]-5-[3-[4-[3-(dimethylamino)prop-l-ynyl]-2-fluoro-phenoxy]propyl]thiazole4-carboxylic acid
Using Deprotection and Hydrolysis General Procedure followed by repurification via reverse phase préparative chromatography (C18, 0.1% TFA in water : MeCN) starting from the product from Step A, the TFA-salt of the desired product was obtained.
HRMS-ESI (m/z): [M+2H]2+ calcd for C39H48FN9O3S2: 386.6647, found 386.6642.
Example 190:2-[[6-(l,3-Benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-[5-[methyl-[2(methy lamino) ethyl] amino] pentyl] amino] -5- [3- [4- [3- (dimethy lamino) prop-1 -yny 1] -2fluoro-phenoxy]propyl]thiazole-4-carboxylic acid
460
Step A: methyl 2-[5-[2-[tert-butoxycarbonyl(methyl)amino]ethyl-methyl-amino]pentyl-[5methyl-6-[(Z)-[3-(2-trimethylsilylethoxymethyl)-l,3-benzothiazol-2ylidene]amino]pyridazin-3-yl]amino]-5-[3-[4-[3-(dimethylamino)prop-l-ynyl]-2-fluoro5 phenoxy]propyl]thiazole-4-carboxylate
Using Alkylation with in situ generated tosylate General Procedure starting from Préparation 5f and tert-butyl N-methyl-N-[2-(methylamino)ethyl]carbamate as the appropriate amine, the desired product was obtained.
Ή NMR (500 MHz, DMSO-ifc) δ ppm 7.84 (d, 1H), 7.66 (s, 1H), 7.47 (d, 1H), 7.43 (td, 1H), 10 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 (br., 2H), 2.73 (s, 3H), 2.45 (s, 3H), 2.35 (br., 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); 13C NMR (125 MHz, DMSO-d6) 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, 15 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-ESI (m/z): [M+H]+ calcd for C51H73FN9O6S2S1: 1018.4873, found 1018.4869.
StepB: 2-[[6-(l,3-benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-[5-[methyl-[2(methylamino)ethyl]amino]pentyl]amino]-5-[3-[4-[3-(dimethylamino)prop-l-ynyl]-2-fluorophenoxy]propyl]thiazole-4-carboxylic acid
Using Deprotection and Hydrolysis General Procedure followed by repurification via reverse phase préparative chromatography (C18, 0.1% TFA in water : MeCN) starting from the product from Step A, the TFA-salt of the desired product was obtained.
HRMS-ESI (m/z): [M+2H]2+ calcd for C39H50FN9O3S2: 387.6726, found 387.6731.
461
Examplel91:2-[5-Aminopentyl-[6-(l,3-benzothiazol-2-ylamino)-5-methyl-pyridazin-3yl]amino]-5-[3-[4-[3-(dimethylamino)prop-l-ynyl]-2-fluoro-phenoxy]propyl]thiazole-4carboxylic acid
Step A: methyl2-[tert-butoxycarbonyl-[5-(tert-butoxycarbonylamino)pentyl]amino]-5-[3-[4[3-(dimethylamino)prop-l-ynyl]-2-fluoro-phenoxy]propyl]thiazole-4-carboxylate
Using Mitsunobu General Procedure starting from 250 mg of Préparation le (0.51 mmol, 1 eq.) and 207 mg of tert-butyl N-(5-hydroxypentyl)carbamate (1.02 mmol, 2 eq.) as the appropriate alcohol, 189 mg (55%) of the desired product was obtained.
Ή NMR (400 MHz, DMSO-d6) 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]+ calcd for C34H50FN4O7S: 677.3, found 677.4.
Step B: methyl 2-[5-(tert-butoxycarbonylamino)pentylamino]-5-[3-[4-[315 (dimethylamino)prop-l-ynyl]-2-fluoro-phenoxy]propyl]thiazole-4-carboxylate
Using Deprotection with HFIP General 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 desired product was obtained.
XH NMR (400 MHz, DMSO-d6) δ ppm 7.56 (t, 1H), 7.30 (dd, 1H), 7.21 (d, 1H), 7.12 (t, 1H), 20 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]+ calcd for C29H42FN4O5S: 577.3, found 577.4.
462
B Step C: methyl 2-[5-(tert-butoxycarbonylamino)pentyl-[5-methyl-6-[(Z)-[3-(2trimethylsilylethoxymethyl)-l,3-benzothiazol-2-ylidene]amino]pyridazin-3-yl]amino]-5-[3[4-[3-(dimethylamino)prop-l-ynyl]-2-fluoro-phenoxy]propyl]thiazole-4-carboxylate
Using Buchwald General Procedure II starting from 102 mg of the product from Step B (0.18 5 mmol, 1 eq.) and 90 mg of Préparation 4a (0.22 mmol, 1.25 eq.) as the appropriate halide, 102 mg (61%) of the desired product was obtained.
>H NMR (500 MHz, DMSO-rA) 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 10 (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); HRMS-ESI (m/z): [M+H]+ calcd for C47H64FN8O6S2Si: 947.4138, found 947.4133.
Step D: 2-[5-aminopentyl-[6-(l,3-benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]amino]5-[3-[4-[3-(dimethylamino)prop-l-ynyl]-2-fluoro-phenoxy]propyl]thiazole-4-carboxylic acid
Using Deprotection and Hydrolysis General Procedure starting from the product from Step C, the desired product was obtained.
HRMS-ESI (m/z): [M+2H]2+ calcd for C35H4iFN8O3S2: 352.1364, found 352.1370.
Example 192:2-r4-Aminobutyl-[6-(l,3-benzothiazol-2-yIamino)-5-methyl-pyridazin-3yl]amino]-5-[3-[4-[3-(dimethylamino)prop-l-ynyl]-2-fluoro-phenoxy]propyl]thiazoIe-420 carboxylic acid
463
Step A: methyl 2-[tert-butoxycarbonyl-[4-(tert-butoxycarbonylamino)butyl]amino]-5-[3~[4[3-(dimethylamino)prop-l-ynyl]-2-fluoro-phenoxy]propyl]thiazole-4-carboxylate
Using Mitsunobu General Procedure starting from 250 mg of Préparation le (0.51 mmol, 1 eq.) and 193 mg of tert-butyl N-(4-hydroxybutyl)carbamate (1.02 mmol, 2 eq.) as the 5 appropriate alcohol, 220 mg (65%) of the desired product was obtained.
Ή NMR (400 MHz, DMSO-r/6) 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-MSESI (m/z): [M+H]+ calcd for C33H48FN4O7S: 663.3, found 663.4.
Step B: methyl 2-[4-(tert-butoxycarbonylamino)butylamino]-5-[3-[4-[3(dimethylamino)prop-l-ynyl]-2-fluoro-phenoxy]propyl]thiazole-4-carboxylate
Using Deprotection with HFIP General Procedure starting from 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.
*H NMR (400 MHz, DMSO-d6) 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]+ calcd for C28H40FN4O5S: 563.3, found 563.2.
Step C: methyl 2-[4-(tert-butoxycarbonylamino)butyl-[5-methyl-6-[(Z)-[3-(220 trimethylsilylethoxymethyl)-l,3-benzothiazol-2-ylidene]amino]pyridazin-3-yl]amino]-5-[3[4~[3-(dimethylamino)prop-l-ynyl]-2-fluoro-phenoxy]propyl]thiazole-4-carboxylate
Using Buchwald General Procedure II starting from 133 mg of the product from Step B (0.24 mmol, 1 eq.) and 120 mg of Préparation 4a (0.29 mmol, 1.25 eq.) as the appropriate halide, 220 mg (98%) of the desired product was obtained.
*H NMR (500 MHz, DMSO-d6) 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); 13C NMR (125 MHz, DMSO-d6) δ ppm 156.1, 128.9, 127.2, 123.5, 123.2, 119.2, 117.6, 115.5, 30 112.0, 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, - .0; HRMS-ESI (m/z): [M+H]+ calcd for C46H62FN8O6S2Si: 933.3982, found 933.3995.
464
Step D: 2-[4-aminobutyl-[6-(l,3-benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]amino]5-[3-[4-[3-(dimethylamino)prop-l-ynyl]-2-fluoro-phenoxy]propyl]thiazole-4-carboxylic acid
Using Deprotection and Hydrolysis General Procedure followed by repurification via reverse phase préparative chromatography (C18, 0.1% TFA in water : MeCN) starting from the 5 product from Step C, the TFA-salt of the desired product was obtained.
HRMS-ESI (m/z): [M+2H]2+ calcd for C34H39FN8O3S2: 345.1280, found 345.1281.
Example 193:2-[[6-(l,3-Benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-[5(dimethylamino)-4-hydroxy-pentyl]amino]-5-[3-[2-fluoro-4-[3-(methylamino)prop-lynyl] phenoxy] propyl] thiazole-4-carboxylic acid
Using Quaternary sait deprotection General Procedure starting from Example 144, Step A, the desired product was obtained.
HRMS-ESI (m/z): [M+H]+ calcd for C36H42FN8O4S2: 733.2749, found 733.2745.
Example 194:2- [ï6-(l,3-Benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl] - [515 (methylamino)pentyl]amino]-5-[3-[4-[3-(dimethylamino)prop-l-ynyl]-2-fluorophenoxy]propyl]thiazole-4-carboxylic acid
465
Step A: methyl5-[3-[4-[3-(dimethylamino)prop-l-ynyl]-2-fluoro-phenoxy]propyl]-2-[5(methylamino)pentyl-[5-methyl-6-[(Z)-[3-(2-trimethylsilylethoxymethyl)-l,3-benzothiazol-2ylidene]amino]pyridazin-3-yl]amino]thiazole-4-carboxylate
Using Alkylation with in situ generated tosylate General Procedure starting from Préparation 5f and methanamine as the appropriate amine, the desired product was obtained after concentration and transferred directly to the next step without any further purification.
Step B: 2-[[6-(l,3-benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-[5(methylamino)pentyl]amino]-5-[3-[4-[3-(dimethylamino)prop-l-ynyl]-2-fluoro10 phenoxy]propyl]thiazole-4-carboxylic acid
Using Deprotection and Hydrolysis General Procedure followed by repurification via reverse phase préparative chromatography (C18, 0.1% TFA in water : MeCN) starting from the crude product from Step A, the TFA-salt of the desired product was obtained.
HRMS-ESI (m/z): [M+H]+ calcd for C36H42FN8O3S2: 717.2800, found 717.2785.
Example 195:2-rr6-(L3-Benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-[5-[3-(4methylpiperazin-l-yl)propylamino]pentyl]amino]-5-[3-[4-[3-(dimethylamino)prop-lynyl]-2-fluoro-phenoxy]propyl]thiazole-4-carboxylic acid
466
Step A: methyl5-[3-[4-[3-(dimethylamino)prop-l-ynyl]-2-fluoro-phenoxy]propyl]-2-[5-[3(4-methylpiperazin-l-yl)propylamino]pentyl-[5-methyl-6-[(Z)-[3-(2trimethylsilylethoxymethyl)-l,3-benzothiazol-2-ylidene]amino]pyridazin-3- yl]amino]thiazole-4-carboxylate
Using Alkylation with in situ generated tosylate General Procedure starting from Préparation 5f and 3-(4-methylpiperazin-l-yl)propan-l-amine as the appropriate amine, the desired product was obtained.
Ή NMR (400 MHz, DMSO-d6) δ ppm 7.83 (dm, 1H), 7.65 (s, 1H), 7.42-7.46 (m, 2H), 7.3610 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]+ calcd for C5oH72FNio04S2Si: 987.5, found 987.4.
Step B: 2-[[6-(l,3-benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-[5-[3-(4methylpiperazin-l-yl)propylamino]pentyl]amino]-5-[3-[4-[3-(dimethylamino)prop-l-ynyl]15 2-fluoro-phenoxy]propyl]thiazole-4-carboxylic acid
Using Deprotection and Hydrolysis General Procedure followed by repurification via reverse phase préparative chromatography (C18, 0.1% TFA in water : MeCN) starting from the product from Step A, the TFA-salt of the desired product was obtained.
HRMS-ESI (m/z): [M+H]+ calcd for C43H56FNio03S2·. 843.3962 found: 843.3943.
Example 196:2-116-(1,3-Benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-[4-hydroxy-5[methyl(2-phosphonoethyl)amino]pentyl]amino]-5-[3-[2-fluoro-4-[3-(methylamino)propl-ynyl]phenoxy]propyl]thiazole-4-carboxylic acid
467
Step A: 2-diethoxyphosphoryl-N-methyl-ethanamine
3.28 g of l-[ethoxy(vinyl)phosphoryl]oxyethane (20 mmol, 1.0 eq.) and 25 mL of methanamine (33w% in EtOH, 200 mmol, 100 eq.) were mixed in a sealed tube and stirred at rt for 20 h. Full conversion was observed. The reaction mixture was concentrated and 3.87 g (99%) of the crude desired product was obtained.
XH NMR (400 MHz, DMSO-dô) 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); 13C NMR (100 MHz, DMSO-î/6) δ ppm 60.9, 60.8, 45.03, 45.01, 35.6, 26.1, 24.7, 16.32, 16.26; GC-TOF (El) (m/z): [M]+ calcd for 10 C7H18NO3P: 195.1024, found 195.1009.
Step B: methyl 5-[3-[4-[3-[tert-butoxycarbonyl(methyl)amino]prop-l-ynyl]-2-fluorophenoxy]propyl]-2-[[4-[tert-butyl(diphenyl)silyl]oxy-5-[2diethoxyphosphorylethyl(methyl)amino]pentyl]-[5-methyl-6-[(Z)-[3-(2trimethylsilylethoxymethyl)-l,3-benzothiazol-2-ylidene]amino]pyridazin-315 yl]amino]thiazole-4-carboxylate
Using Alkylation with tosylate General Procedure starting from Préparation 5a and the product from Step A as the appropriate amine, the desired product was obtained.
HRMS-ESI (m/z): [M+H]+ calcd for C69H95FN8OioPS2Si2: 1365.5867, found 1365.5916.
Step C: 2-][6-(l,3-benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-[4-hydroxy-520 [methyl(2-phosphonoethyl)amino]pentyl]amino]-5-[3-]2-fluoro-4-[3-(methylamino)prop-lynyl]phenoxy]propyl]thiazole-4-carboxylic acid
Using Deprotection and hydrolysis of phosphonic acid dérivatives General Procedure starting from the product from Step B, the desired product was obtained.
HRMS-ESI (m/z): [M+H]+ calcd for C37H45FN8O7PS2: 827.2569, found 827.2561.
468
Example 197:2-({6-[(l,3-Benzothiazol-2-yl)amino]-5-methylpyridazin-3yl}(methyl)amino)-5-[3-({4-fluoro-6-[3-(methylamino)prop-l-yn-l-yl]pyridin-3yl}oxy)propyl] -l,3-thiazole-4-carboxylic acid
Step A: 2-chloro-4-fluoro-5-(4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2-yl)pyridine
To a solution of 2-chloro-4-fluoropyridine (2.5 g; 19.01 mmol) in tetrahydrofuran (30 mL) was successively added 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 (l,5-cyclooctadiene)(methoxy)iridium(I) dimer (63.0 mg; 0.095 mmol; 0.005 eq.). The reaction mixture was flushed with Argon and was stirred at 80 °C for 16 h and then concentrated to dryness. The resulting residue was purified by column chromatography on silica gel using cyclohexane/ethyl acetate as eluent to afford the desired compound (3.64 g; 74%).
XH NMR (DMSO-de) 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-fluoro-pyridin-3-ol
To a solution of the product from Step A (2.00 g; 7.77 mmol) in tetrahydrofuran (16 mL) was successively added at 0°C, sodium hydroxide (311 mg; 7.77 mmol; 1.0 eq.) and an aqueous solution of hydrogen peroxide 33% (2.38 mL; 23.3 mmol; 3.0 eq.). The reaction mixture was stirred at 0°C for 3 h and at rt for 16 h. The reaction mixture was diluted with water (50 mL) and was acidified with an aqueous solution of hydrogen chloride IN until pH = 1. The aqueous layer was extracted with dichloromethane (3 x 100 mL) and the combined organic layers were dried over sodium sulfate, filtered, and concentrated to dryness. The resulting residue was purified by column chromatography on silica gel using cyclohexane/ethyl acetate as eluent to afford the desired compound (640 mg; 56%).
XH NMR (DMSO-r/ό) δ ppm 7.53 (d, 1H, J = 10.4 Hz), 8.06 (d, 1H, J = 10.8 Hz), 10.76 (s, 1H).
469
Step C: tert-butyl N-[3-(4-fluoro-5-hydroxy-2-pyridyl)prop-2-ynyl]-N-methyl-carbamate
To a solution of the product from Step B (300 mg; 2.03 mmol) in acetonitrile (12 mL) was successively added tert-butyl Wmethyl-JV-prop-2-ynyl-carbamate (688 mg; 4.07 mmol; 2.0 eq.), césium 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 flushed with argon and was stirred for 16 h at 70 °C. The reaction mixture was diluted with water (50 ml.) and was acidified with an aqueous solution of hydrogen chloride IN until pH - 1. The aqueous layer was extracted with dichloromethane (3 x 100 mL) and the combined organic layers were dried over sodium sulfate, filtered and concentrated to dryness. The resulting residue was purified by column chromatography on silica gel using cyclohexane/ethyl acetate as eluent to afford the desired compound (257 mg; 45%).
XH 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-(l,3-benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-methyl-amino]-5[3-[[6-[3-[tert-butoxycarbonyl(methyl) amino]prop-l -ynyl]-4-fluoro-3pyridyl]oxy]propyl]thiazole-4-carboxylate
Using Alkylation General Procedure starting from Préparation 5g and the product from Step C as the appropriate phénol, the desired compound was obtained.
Ή NMR (500 MHz, DMSO-d6) 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); 13C NMR (125 MHz, DMSO-d6) 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; HRMS-ESI (m/z): [M+H]+ calcd for C36H40FN8O5S2: 747.2541, found 747.2526.
Step E: 2-({6-[(l,3-Benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)-5-[3({4-fluoro-6-[3-(methylamino)prop-l-yn-l-yl]pyridin-3-yl}oxy)propyl]-l,3-thiazole-4carboxylic acid
470
Using Deprotection and Hydrolysis General Procedure starting from Step D as the appropriate ethyl ester, the desired product was obtained.
Hl NMR (500 MHz, DMSO-d6) 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); 13C NMR (125 MHz, DMSO-dô) 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; HRMS-ESI (m/z): [M+H]+ calcd for C29H28FN8O3S2: 619.1704, found 619.1698.
Example 198:2-[[6-(l,3-BenzothiazoI-2-ylamino)-5-methyl-pyridazin-3-yI]-methylamino]-5-[3-[2-fluoro-4-(3-morpholinoprop-l-ynyl)phenoxy]propyl]thiazole-4carboxylic acid
Step A: ethyl 5-[3-[2-fluoro-4-(3-morpholinoprop-l-ynyl)phenoxy]propyl]-2-[methyl-[5methyl-6-[ (Z) -[3-(2-trimethylsilylethoxy methyl) -l,3-benzothiazol-2ylidene]amino]pyridazin-3-yl]amino]thiazole-4-carboxylate
Using Alkylation General Procedure starting from Préparation 5g and Préparation 6e as the appropriate phénol, the desired product was obtained.
XH NMR (400 MHz, DMSO-d6) 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); 13C NMR (100 MHz, DMSO-dô) 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; HRMS-ESI (m/z): [M+H]+ calcd for
471 φ C41H51FN7O5S2S1: 832.3141, found 832.3146.
Step B: 2-[[6-(l,3-benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-methyl-amino]-5-[3-[2fluoro-4-(3-niorpholinoprop-l-ynyl)phenoxy]propyl]thiazole-4-carboxylic acid
Using Deprotection and Hydrolysis General Procedure starting from the product from Step
A as the appropriate ethyl ester, the desired product was obtained.
HRMS-ESI (m/z): [M+H]+ calcd for C33H33FN7O4S2: 674.2014, found 674.2006.
Example 199:2-[[6-(l,3-Benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-[6(dimethylamino)hexyl]amino]-5-[3-[2-fluoro-4-[3-(methylamino)prop-lynyl] phenoxy] propyl] thiazole-4-carboxy lie acid
Step A: methyl2-[tert-butoxycarbonyl-[6-(dimethylamino)hexyl]amino]-5-[3-[4-[3-[tertbutoxycarbonyl(methyl)amino]prop-l-ynyl]-2-fluoro-phenoxy]propyl]thiazole-4carboxylate
Using Mitsunobu General Procedure starting from Préparation 1b and 615 (dimethylamino)hexan-l-ol, 1.51 g (quant., contained ca. 40n/n% DIAD-2H) of the desired product was obtained.
XH NMR (400 MHz, DMSO-î/6) 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), 1.701.20 (m, 8H), 1.61 (m, 2H), 1.50/1.41 (s, 18H); HRMS-ESI (m/z): [M+H]+ calcd for 20 C36H54FN4O7S: 705.3697, found 705.3699.
Step B: methyl 5-[3-[4-[3-[tert-butoxycarbonyl(methyl)amino]prop-l-ynyl]-2-fluorophenoxy]propyl]-2-[6-(dimethylamino)hexylamino]thiazole-4-carboxylate
472
Using Deprotection with HFIP General Procedure starting from the product from Step A,
650 mg (50%) of the desired product was produced.
’H NMR (500 MHz, DMSO-d6) 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); 13C NMR (125 MHz, DMSO-d6) 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; HRMS-ESI (m/z): [M+H]+ calcd for C31H46FN4O5S: 605.3173, found 605.3160.
Step C: methyl 5-[3-[4-[3-[tert-butoxycarbonyl(methyl)amino]prop-l-ynyl]-2-fluorophenoxy]propyl]-2-[6-(dimethylamÎno)hexyl-[5-methyl-6-[(Z)-[3-(2trimethylsilylethoxymethyl)-l,3-benzothiazol-2-ylidene]amino]pyridazin-3yl]amino]thiazole-4-carboxylate
Using Buchwald General Procedure II starting from the product from Step B and Préparation 4a, 1.06 g (98%) of the desired product was produced.
XH NMR (500 MHz, DMSO4) 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); 13C NMR (125 MHz, DMSO-t/6) 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; HRMS-ESI (m/z): [M+H]+ calcd for C49H68FN8O6S2Si: 975.4457, found 975.4437.
Step D: 2-[[6-(l,3-benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-[6(dimethylamino)hexyl]amino]-5-[3-[2-fluoro-4-[3-(methylamino)prop-lynyl]phenoxy]propyl]thiazole-4-carboxylic acid
Using Deprotection and Hydrolysis General Procedure followed by repurification via reverse phase préparative chromatography (C18, 0.1% TFA in water : MeCN) starting from the product from Step C, the TFA-salt of the desired product was obtained.
HRMS-ESI (m/z): [M+H]+ calcd for C37H44FN8O3S2: 731.2962, found 731.2968.
473
Example 200:2-[[6-(l,3-Benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-[3(dimethylamino)propyl]amino]-5-[3-[2-fluoro-4-[3-(methylamino)prop-lyny 1] phenoxy] propyl] thiazole-4-carboxylie acid
Step A: methyl 2-[tert-butoxycarbonyl-[3-(dimethylamino)propyl]amino]-5-]3-[4-[3-[tertbutoxycarbonyl(methyl)amino]prop-l-ynyl]-2-fluoro-phenoxy]propyl]thiazole-4carboxylate
Using Mitsunobu General Procedure starting from Préparation 1b and 3(dimethylamino)propan-l-ol, 1.40 g (quant., the sample contained aprox. 35 n/n% DIAD-2H) 10 of the desired product was produced.
XH NMR (400 MHz, DMSO-c/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); HRMS-ESI (m/z): [M+H]+ calcd for C33H48FN4O7S: 663.3228, found 663.3218.
Step B: methyl 5-]3-[4-[3-[tert-butoxycarbonyl(methyl)amino]prop-l-ynyl]-2-fluorophenoxy]propyl]-2-[3-(dimethylamino)propylamino]thiazole-4-carboxylate
Using Deprotection with HFIP General Procedure starting from the product from Step A, 0.95 g (80%) of the desired product was produced.
‘H NMR (500 MHz, DMSO-t/6) δ ppm 7.57 (t, 1H), 7.31 (d, 1H), 7.21 (d, 1H), 7.13 (t, 1H), 20 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), .11 (s, 6H), 2.00 (quint., 2H), 1.63 (m, 2H), 1.41 (s, 9H); 13C NMR (125 MHz, DMSO-d6) 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; HRMSESI (m/z): [M+H]+ calcd for C28H40FN4O5S: 563.2703, found 563.2694.
Step C: methyl 5-[3-[4-[3-[tert-butoxycarbonyl(methyl)amino]prop-1 -ynyl]-2-fluoro20487
474 phenoxy]propyl]-2-[3-(dimethylamino)propyl-[5-methyl-6-[(Z)-[3-(2trimethylsilylethoxymethyl)-l,3-benzothiazol-2-ylidene]amino]pyridazin-3yl]amino]thiazole-4-carboxylate
Using Buchwald General Procedure II starting from the product from Step B and 5 Préparation 4a, 0.79 g (51%) of the desired product was produced.
Tl NMR (500 MHz, DMSO-zL) 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); 13C 10 NMR (125 MHz, DMSO-d6) 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, 25.0, 23.1, 17.9, 17.8, -1.0; HRMS-ESI (m/z): [M+H]+ calcd for C46H62FNsO6S2Si: 933.3987, found 933.3990.
Step D: 2-[[6-(l,3-benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-[3(dimethylamino)propyl]amino]-5-[3-[2-fluoro-4-[3-(methylamino)prop-l 15 ynyl]phenoxy]propyl]thiazole-4-carboxylic acid
Using Deprotection and Hydrolysis General Procedure followed by repurification via reverse phase préparative chromatography (C18, 0.1% TFA in water : MeCN) starting from the product from Step C, the TFA-salt of the desired product was obtained.
HRMS-ESI (m/z): [M+2H]2+ calcd for C34H39FN8O3S2: 345.1280, found 345.1265.
Example 201:2-116-(1,3-Benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-[2(trimethylammonio)ethyl]amino]-5-[3-[2-fluoro-4-[3-(methylamino)prop-lynyl] phenoxy] propyl] thiazole-4-carboxylate
475
Using Alkylation, Deprotection and Hydrolysis General procedure starting from Example
207 , Step C and iodomethane, the TFA-salt of the desired product was obtained.
HRMS-ESI (m/z): [M+2H]2+ calcd for C34H39FN8O3S2: 345.1280, found 345.1279.
Example 202; 2-[[6-(l,3-Benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-[35 (trimethylammonio)propyl]amino]-5-[3-[2-fluoro-4-[3-(methylamino)prop-ly ny 1] phenoxy] propyl] thiazole-4-carboxy late
Using Alkylation, Deprotection and Hydrolysis General procedure starting from Example 200, Step C and iodomethane, the TFA-salt of the desired product was obtained.
HRMS-ESI (m/z): [M+2H]2+ calcd for C35H41FN8O3S2: 352.1358, found 352.1365.
Example 203:2-rr6-(l,3-Benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-methylamino]-5-[3-[2-fluoro-4-(3-pyrrolidin-l-ylprop-l-ynyl)phenoxy]propyl]thiazole-4carboxylic acid
476
Step A: ethyl 5-[3-[2-fluoro-4-(3-pyrrolidin-l-ylprop-l-ynyl)phenoxy]propyl]-2-[methyl-[5methyl-6-[(Z)-[3-(2-trimethylsilylethoxy methyl)-l,3-benzothiazol-2ylidene]amino]pyridazin-3-yl]amino]thiazole-4-carboxylate
Using Alkylation General Procedure starting from Préparation 5g and Préparation 6c as 5 the appropriate phénol, the desired product was obtained.
XH NMR (400 MHz, DMSO-d6) 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, 2H), -0.11 (s, 9H); HRMS-ESI (m/z): [M+H]+ 10 calcd for C4iH5iFN7O4S2Si: 816.3192, found 816.3189.
Step B: 2-[[6-(l,3-benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-methyl-amino]-5-[3-[2fluoro-4-(3-pyrrolidin-l-ylprop-1 -ynyl)phenoxy]propyl]thiazole-4-carboxylic acid
Using Deprotection and Hydrolysis General Procedure starting from the product from Step A as the appropriate ethyl ester, the desired product was obtained.
HRMS-ESI (m/z): [M+H]+ calcd for C33H33FN7O3S2: 658.2065, found 658.2060.
Example 204:2-[[6-(l,3-Benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-methylamino]-5-[3-[2-fluoro-4-[3-(l-piperidyl)prop-l-ynyl]phenoxy]propyl]thiazole-4carboxylic acid
Step A: ethyl5-[3-[2-fluoro-4-[3-(l-piperidyl)prop-l-ynyl]phenoxy]propyl]-2-[methyl-[5methyl-6-[(Z)-[3-(2-trimethylsilylethoxymethyl)-l,3-benzothiazol-2ylidene]amino]pyridazin-3-yl]amino]thiazole-4-carboxylate
Using Alkylation General Procedure starting from Préparation 5g and Préparation 6d as
477
P the appropriate phénol, the desired product was obtained.
Ή NMR (400 MHz, DMSO-d6) 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 5 (m, 2H), 1.50 (m, 4H), 1.35 (m, 2H), 1.29 (t, 3H), 0.92 (m, 2H), -0.12 (s, 9H); HRMS-ESI (m/z): [M+H]+ calcd for C42H53FN7O4S2Si: 830.3348, found 830.3347.
Step B: 2-[[6-(l,3-benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-methyl-aniino]-5-[3-[2fluoro-4-[3-(l-piperidyl)prop-l-ynyl]phenoxy]propyl]thiazole-4-carboxylic acid
Using Deprotection and Hydrolysis General Procedure starting from the product from Step 10 A as the appropriate ethyl ester, the desired product was obtained.
HRMS-ESI (m/z): [M+H]+ calcd for C34H35FN7O3S2: 672.2221, found 672.2217.
Example 205:2-[[6-(l,3-Benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-methylamino]-5-[3-[4-[3-(dimethylamino)but-l-ynyl]-2-fluoro-phenoxy]propyl]thiazole-4carboxylic acid
Step A : ethyl 5-[3-[4-[3-(dimethylamino)but-l-ynyl]-2-fluoro-phenoxy]propyl]-2-[methyl[5-methyl-6-[(Z)-[3-(2-trimethylsilylethoxymethyl)-l,3-benzothiazol-2ylidene]amino]pyridazin-3-yl]amino]thiazole-4-carboxylate
Using Alkylation General Procedure starting from Préparation 5g and Préparation 6f as 20 the appropriate phénol, the desired product was obtained.
Step B: 2-[[6-(l,3-benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-methyl-amino]-5-[3-[4[3-(dimethylamino)but-l-ynyl]-2-fluoro-phenoxy]propyl]thiazole-4-carboxylic acid
478
Using Deprotection and Hydrolysis General Procedure starting from the product from Step
A as the appropriate ethyl ester, the desired product was obtained.
HRMS-ESI (m/z): [M+H]+ calcd for C32H33FN7O3S2: 646.2065, found 646.2057.
Example 206:2-[[6-(l,3-Benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-methylamino]-5-[3-[2-fluoro-4-((3R or 3S)-morpholinobut-l-ynyl)phenoxy]propyl]thiazole-4carboxylic acid (enantiomer 2)
Step A: ethyl 5-[3-[2-fluoro-4-((3R or 3S)-morpholinobut-l-ynyl)phenoxy]propyl]-2[methyl-[5-methyl-6-[ (Z) -[3-(2-trimethylsilylethoxymethyl) -l,3-benzothiazol-2ylidene]amino]pyridazin-3-yl]amino]thiazole-4-carboxylate (enantiomer 2)
Using Alkylation General Procedure starting from Préparation 5g and Préparation 6h as the appropriate phénol, the desired product was obtained.
Ή NMR (400 MHz, DMSO-d6) 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); 13C NMR (100 MHz, DMSO-d6) 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, 23.2, 19.3, 17.8, 17.8, 14.6, -1.0; HRMS-ESI (m/z): [M+H]+ calcd for C42H53FN7O5S2Si: 846.3297, found 846.3283.
Step B: 2-[[6-(l,3-benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-methyl-amino]-5-[3-[2fluoro-4-((3R or 3S)-morpholinobut-l-ynyl)phenoxy]propyl]thiazole-4-carboxylic acid (enantiomer 2)
Using Deprotection and Hydrolysis General Procedure starting from the product from Step A as the appropriate ethyl ester, the desired product was obtained.
479
HRMS-ESI (m/z): [M+H]+ calcd for C34H35FN7O4S2: 688.2176, found 688.2171.
Example 207:2-[[6-(l,3-Benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-[2(dimethyIamino)ethyl]amino]-5-[3-[2-fluoro-4-[3-(methylamino)prop-lyny 1] phenoxy] propyl] thiazole-4-carboxylic acid /
—N
OH
Step A : methyl 2-[tert-butoxycarbonyl-[2-(dimethylamino)ethyl]amino]-5-[3-[4-[3-[tertbutoxycarbonyl(methyl)amino]prop-l-ynyl]-2-fluoro-phenoxy]propyl]thiazole-4carboxylate
Using Mitsunobu General Procedure starting from Préparation 1b, and 2(dimethylamino)éthanol, 0.90 g (70%) of the desired product was produced.
XH NMR (500 MHz, DMSO-rA) 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); 13C NMR (125 MHz, DMSO-d0) 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; HRMS-ESI (m/z): [M+H]+ calcd for C32H46FN4O7S: 649.3071, found 649.3059.
Step B: methyl 5-[3-[4-[3-[tert-butoxycarbonyl(methyl)amino]prop-l-ynyl]-2-fluorophenoxy]propyl]-2-[2-(dimethylamino)ethylamino]thiazole-4-carboxylate
Using Deprotection with HFIP General Procedure starting from the product from Step A, 0.68 g (89%) of the desired product was produced.
XH NMR (500 MHz, DMSO-ί/ό) 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); X3C 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; HRMS-ESI (m/z):
480
Φ [Μ+Η]+ calcd for C27H38FN4O5S: 549.2547, found 549.2543.
Step C: methyl 5-[3-[4-[3-[tert-butoxycarbonyl(methyl)amino]prop-l-ynyl]-2-fluorophenoxy]propyl]-2-[2-(dimethylamino)ethyl-[5-methyl-6-[(Z)-[3-(2trimethylsilylethoxymethyl)-l,3-benzothiazol-2-ylidene]amino]pyridazin-35 yl]amino]thiazole-4-carboxylate
Using Buchwald General Procedure II starting from the product from Step B and Préparation 4a, 0.57 g (50%) of the desired product was produced.
Ή NMR (500 MHz, DMSO-d6) δ ppm 7.84 (d, 1Η), 7.66 (s, 1Η), 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), 10 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), .23 (s, 6H), 2.12 (m, 2H), 1.40 (s, 9H), 0.92 (t, 2H), -0.11 (s, 9H); 13C NMR (125 MHz, DMSO-dô) 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; HRMS-ESI (m/z): [M+H]+ calcd for C45H6oFN806S2Si: 919.3831, found 919.3829.
Step D: 2-[[6-(l,3-benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-[2(dimethylamino)ethyl]amino]-5-[3-[2-fluoro-4-[3-(methylamino)prop-lynyl]phenoxy]propyl]thiazole-4-carboxylic acid
Using Deprotection and Hydrolysis General Procedure followed by repurification via reverse phase préparative chromatography (C18, 0.1% TFA in water : MeCN) starting from the 20 product from Step C, the TFA-salt of the desired product was obtained.
HRMS-ESI (m/z): [M+H]+ calcd for C33H36FN8O3S2: 675.2336, found 675.2323.
Example 208:2-[[6-(l,3-Benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-methylamino]-5-[3-[2-fluoro-4-((3S or 3R)-morpholinobut-l-ynyl)phenoxy]propyl]thiazole-4carboxylic acid (enantiomer 1)
481
Step A: ethyl 5-[3-[2-fluoro-4-((3S or 3R)-morpholinobut-l-ynyl)phenoxy]propyl]-2[methyl-[5-methyl-6-[(Z)-[3-(2-trimethylsilylethoxymethyl)-l,3-benzothiazol-2ylidene]amino]pyridazin-3-yl]amino]thiazole-4-carboxylate (enantiomer 1)
Using Alkylation General Procedure starting from Préparation 5g and Préparation 6g as the appropriate phénol, the desired product was obtained.
Ή NMR (400 MHz, DMSO-d6) 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 10 (s, 3H), 2.13 (quint., 2H), 1.30 (t, 3H), 1.28 (d, 3H), 0.92 (t, 2H), -0.11 (s, 9H); 13C NMR (100
MHz, DMSO-d6) 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, 23.2, 19.3, 17.8, 17.8, 14.6, -1.0; HRMS-ESI (m/z): [M+H]+ calcd for C42H53FN7O5S2Sr. 846.3297, found 846.3288.
Step B: 2-[[6-(l,3-benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-methyl-amino]-5-[3-[215 fluoro-4-((3S or 3R)-morpholinobut-l -ynyl)phenoxy]propyl]thiazole-4-carboxylic acid (enantiomer 1)
Using Deprotection and Hydrolysis General Procedure starting from the product from Step A as the appropriate ethyl ester, the desired product was obtained.
HRMS-ESI (m/z): [M+H]+ calcd for C34H35FN7O4S2·. 688.2170, found 688.2166.
Example 209:2-iï6-(l,3-Benzothiazol-2-ylamino)-5-(trifluoromethyl)pyridazin-3-yl]methyl-amino]-5-[3-[2-fluoro-4-[3-(methylamino)prop-l-ynyl]phenoxy]propyl]thiazole4-carboxylic acid
482
Step A: methyl 2-[[6-chloro-5-(trifluoromethyl)pyridazin-3-yl]-methyl-amino]-5-[3-(2fluoro-4-iodo-phenoxy)propyl] thiazole-4-carboxylate
A 50 mL oven-dried, one-necked, round-bottomed flask equipped with a PTFE-coated 5 magnetic stirring bar and fitted with a reflux condenser. It was charged with 991 mg of
Préparation 3s (2.20 mmol, 1.1 eq.) dissolved in 10 mL of dry THF then it was cooled to 0°C and 96 mg of NaH (2.40 mmol, 1.2 eq., 60% in minerai oil) was added portionwise over a period of 5 min. After stirring for 15 min at 0°C 434 mg of 3,6-dichloro-4(trifluoromethyl)pyridazine (2 mmol, 1 eq.) was added in one portion, and stirred at that 10 température for 1 h, when the reaction reached full conversion. The reaction mixture was quenched with the addition of cc. NH4CI solution and was extracted with EtOAc. The combined organic layers were dried over Na2SÜ4, 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. It was purified via flash column chromatography using heptane and 15 EtOAc as eluents to give 500 mg (39%) of the desired product as yellow solid.
*H NMR (500 MHz, DMSO-d6) 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); 13C NMR (125 MHz, DMSO-d6) δ ppm 162.7, 156.2, 155.3, 152.2, 147.0, 144.2, 143.2, 135.1, 134, 128.5, 124.9, 121.4, 117.9, 117.6, 82.4, 68.2, 52.1, 35.9, 30.6, 23.5; HRMS-ESI (m/z): [M+H]+ calcd for 20 C20H17CIF4IN4O3S: 630.9685, found 630.9679.
Step B: methyl 5-[3-[4-[3-[tert-butoxycarbonyl(methyl)amino]prop-l-ynyl]-2-fluorophenoxy]propyl]-2-[[6-chloro-5-(trifluoromethyl)pyridazin-3-yl]-methyl-amino]thiazole-4carboxylate
A 12 mL oven-dried vial equipped with a PTFE-coated magnetic stirring bar was charged with 25 330 mg of the product from Step A (0.52 mmol, 1 eq.), 177 mg of tert-butyl N-methyl-N-prop2-ynyl-carbamate (1.04 mmol, 2 eq.), 2.5 mL of dry THF and 0.5 mL of DIPA then placed
483 under an inert atmosphère through a gas inlet. After addition of 11.5 mg of Pd(PPh3)2C12 (0.026 mmol, 0.05 eq.) and 4.98 mg of Cul (0.026 mmol, 0.05 eq.) the reaction mixture was warmed up to 60°C and stirred at that température for 1 h, when the reaction reached full conversion. Celite was added to the reaction mixture and the volatiles were removed under reduced pressure. Then it was purified via flash column chromatography using heptane and EtOAc to give 271 mg (69%) of the desired product.
Ή NMR (500 MHz, DMSO-d6) 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); 13C NMR (125 MHz, DMSO-d6) 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-ESI (m/z): [M+H]+ calcd for C29H3iClF4N5O5S: 672.1665, found 672.1651.
Step C: methyl 2-[[6-(l,3-benzothiazol-2-ylamino)-5-(trifluoromethyl)pyridazin-3-yl]methyl-amino]-5-[3-[4-[3-[tert-butoxycarbonyl(methyl)amino] prop-1 -ynyl]-2-fluorophenoxy]propyl]thiazole-4-carboxylate
A 4 mL oven-dried vial equipped with a PTFE-coated magnetic stirring bar, was charged with 130 mg of the product from Step B (0.19 mmol, 1 eq.), 87 mg of l,3-benzothiazol-2-amine (0.58 mmol, 3 eq.) and 168 uL of DIPEA (125 mg, 0.96 mmol, 5 eq.), suspended in 1 mL CyOH and then placed under an inert atmosphère flushed with argon. After 2 min stirring under inert atmosphère 18 mg of Pdz(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 warmed up to 140°C and stirred at that température for 4 h, when the reaction reached full conversion. The reaction mixture was diluted with 1 mL DCM and purified via flash column chromatography using heptane and EtOAc as eluents to give 85 mg (56%) of the desired product.
Hl NMR (500 MHz, DMSO-d6) 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); 13C NMR (125 MHz, DMSO-dô) 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; HRMSESI (m/z): [M+H]+ calcd for C36H36F4N7O5S2: 786.2150, found 786.2148.
Step D: 2-[[6-(l,3-benzothiazol-2-ylamino)-5-(trifluoromethyl)pyridazin-3-yl]-methylamino]-5-[3-[2-fluoro-4-[3-(methylamino)prop-l-ynyl]phenoxy]propyl]thiazole-4carboxylic acid
484
Using Deprotection and Hydrolysis General Procedure starting from the product from Step C as the appropriate methyl ester, the desired product was obtained.
HRMS-ESI (m/z): [M+H]+ calcd for C30H26F4N7O3S2: 672.1469, found 672.1458.
Example 210:2-[[6-(l,3-Benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-[45 (dimethylamino)butyl]amino]-5-[3-[2-fluoro-4-[3-(methylamino)prop-lyny 1] phenoxy] propy 1] thiazole-4-carboxylie acid
Step A: methyl 2-[tert-butoxycarbonyl-[4-(dimethylamino)butyl]amino]-5-[3-(2-fluoro-4iodo-phenoxy)propyl]thiazole-4-carboxylate
Using Mitsunobu General Procedure starting from Préparation la and 4(dimethylamino)butan-l-ol, 401 mg (quant.) of the desired product was obtained.
’H NMR (400 MHz, DMSO-dô) 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); 13C NMR (100 MHz, DMSO-dô) δ ppm 162.8, 152.7, 147.0, 143.6, 15 135.5, 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, 21.7;
HRMS-ESI (m/z): [M+H]+ calcd for C25H36FIN3O5S: 636.1399, found 636.1400.
Step B: methyl 5-[3-[4-[3-[tert-butoxycarbonyl(methyl)amino]prop-1-ynyl]-2-fluorophenoxy]propyl]-2-[4-(dimethylamino)butylamino]thiazole-4-carboxylate
Using Deprotection with HFIP General Procedure starting from the product from Step A, 20 crude methyl 2-[4-(dimethylamino)butylamino]-5-[3-(2-fluoro-4-iodophenoxy)propyl]thiazole-4-carboxylate (LC-MS-ESI (m/z): [M+H]+ calcd for C20H28FIN3O3S: 536.1, found 536.2) was obtained after concentration then Sonogashira General Procedure was applied with tert-butyl N-methyl-N-prop-2-ynyl-carbamate as alkyne reactant which was
485 resulted in 240 mg (67%) of the desired product.
ΧΗ NMR (500 MHz, DMSO-d6) 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); 13C NMR (125 MHz, 5 DMSO-dô) 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; HRMS-ESI (m/z): [M+H]+ calcd for C29H42FN4O5S: 577.2854, found 577.2854.
Step C: methyl 5-[3-[4-[3-[tert-butoxycarbonyl(methyl)amino]prop-l-ynyl]-2-fluorophenoxy]propyl]-2-[4-(dimethylamino)butyl-[5-methyl-6-[(Z) -[3-(2trimethylsilylethoxymethyl)-l,3-benzothiazol-2-ylidene]amino]pyridazin-310 yl]amino]thiazole-4-carboxylate
Using Buchwald General Procedure II starting from 235 mg of the product from Step B (0.41 mmol, 1 eq.) and 249 mg of Préparation 4a (0.61 mmol, 1.5 eq.) as the appropriate halide, 353 mg (90%) of the desired product was obtained.
'H NMR (500 MHz, DMSO-d6) δ ppm 7.84 (d, 1H), 7.73 (s, 1H), 7.47 (dm, 1H), 7.44 (t, 1H), 15 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); 13C NMR (125 MHz, DMSO-d6) 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, 20 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;
HRMS-ESI (m/z): [M+H]+ calcd for C47H64FN8O6S2Si: 947.4138, found 947.4144.
Step D: 2-[[6-(l,3-benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-[4(dimethylamino)butyl]amino]-5-[3-[2-fluoro-4-[3-(methylamino)prop-lynyl]phenoxy]propyl]thiazole-4-carboxylic acid
Using Deprotection and Hydrolysis General Procedure followed by repurification via reverse phase préparative chromatography (C18, 0.1% TFA in water : MeCN), starting from the product from Step C, the TFA-salt of the desired product was obtained.
HRMS-ESI (m/z): [M+H]+ calcd for C35H4oFN803S2: 703.2649, found 703.2637.
486
Example 2ll:2-rr6-(l.3-Benzothiazol-2-vlamino)-5-methvl-pvridazin-3-vI1-r4-hvdroxv-5(trimethylammonio)pentyl]amino]-5-[3-[4-[3-(dimethylamino)prop-l-ynyl]-2-fluorophenoxy] propyl] thiazole-4-carboxylate
StepA: [2-[tert-butyl(diphenyl)silyl]oxy-5-[[5-[3-[4-[3-(dimethylamino)prop-l-ynyl]-2fluoro-phenoxy]propyl]-4-methoxycarbonyl-thiazol-2-yl]-[5-methyl-6-[(Z)-[3-(2trimethylsilylethoxymethyl)-l,3-benzothiazol-2-ylidene] amino]pyridazin-3yl]amino]pentyl]-trimethyl-ammonium
Using Quaternary sait formation General Procedure starting from Préparation 5b and N,N10 dimethylmethanamine as the appropriate tertiary amine, the desired product was obtained.
Step B: 2-[[6-(l,3-benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-[4-hydroxy-5(trimethylammonio)pentyl]amino]-5-[3-[4-[3-(dimethylamino)prop-l-ynyl]-2-fluorophenoxy]propyl]thiazole-4-carboxylate
Using Quaternary sait deprotection General Procedure starting from the product from Step 15 A as the quaternary sait, the desired product was obtained.
HRMS-ESI (m/z): [M+H]+ calcd for C38H46FN8O4S2: 761.3062, found 761.3057.
Example 212:3-15-ri6-(1.3-Benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-[4carboxy-5-[3-[2-fluoro-4-[3-(methylamino)prop-l-ynyl]phenoxy]propyl]thiazol-2y I] amino] penty 1-dimethyl-ammonio] propane- 1-sulfonate
487
Using Alkylation, Deprotection and Hydrolysis General procedure starting from Example 213, Step C and oxathiolane 2,2-dioxide, the TFA-salt of the desired product was obtained.
HRMS-ESI (m/z): [M+H]+ calcd for CsiHTzFNgOgSsSi: 839.2843, found 839.2823.
Example 213:2-[T6-(1.3-Benzothiazol-2-vlamino)-5-methyl-pyridazin-3-yl]-[5(dimethylamino)pentyl]amino]-5-[3-[2-fluoro-4-[3-(methylamino)prop-lynyl]phenoxy]propyl]thiazole-4-carboxylic acid
Step A: methyl 2-[tert-butoxycarbonyl-[5-(dimethylamino)pentyl]amino]-5-[3-[4-[3-[tert10 butoxycarbonyl(methyl)amino]prop-l-ynyl]-2-fluoro-phenoxy]propyl]thiazole-4carboxylate
Using Mitsunobu General Procedure starting from Préparation 1b and 5(dimethylamino)pentan-l-ol, 1.47 g (quant.) of the desired product was obtained.
Ή NMR (500 MHz, DMSO-î/6) 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); 13C NMR (125 MHz, DMSO-î/6) 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,
488
I 23.2; HRMS-ESI (m/z): [M+H]+ calcd for C35H52FN4O7S: 691.3541, found 691.3524.
Step B: methyl 5-[3-[4-[3-[tert-butoxycarbonyl(methyl)amino]prop-l-ynyl]-2-fluorophenoxy]propyl]-2-[5-(dimethylamino)pentylamino]thiazole-4-carboxylate
Using Deprotection with HFIP General Procedure starting from the product from Step A, 5 0.66 g (52%) of the desired product was produced.
*Η NMR (500 MHz, DMSO-d6) 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); 13C NMR (125 MHz, DMSO-t/6) δ ppm 129.1, 119.3, 115.4, 68.0, 58.9, 51.7, 45.0, 44.4, 38.6, 10 33.8, 30.6, 28.9, 28.5, 26.4, 24.5, 23.3; HRMS-ESI (m/z): [M+H]+ calcd for C30H44FN4O5S:
591.3016, found 591.3005.
Step C: methyl 5-[3-[4-[3-[tert-butoxycarbonyl(methyl)amino]prop-l-ynyl]-2-fluorophenoxy]propyl]-2-[5-(dimethylamino)pentyl-[5-methyl-6-[(Z) -[3-(2trimethylsilylethoxymethyl)-l,3-benzothiazol-2-ylidene]amino]pyridazin-3- yl]amino]thiazole-4-carboxylate
Using Buchwald General Procedure II starting from the product from Step B, and Préparation 4a, 0.43 g (40%) of the desired product was produced.
*H NMR (500 MHz, DMSO-tfo) 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., 20 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); 13C NMR(125 MHz, DMSO-ώ,) 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; HRMS-ESI (m/z): [M+H]+ calcd for C48H66FN8O6S2Si: 961.4300, 25 found 461.4293.
Step D: 2-[[6-(l,3-benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-[5(dimethylamino)pentyl]amino]-5-[3-[2-fluoro-4-[3-(methylamino)prop-lynyl]phenoxy]propyl]thiazole-4-carboxylic acid
Using Deprotection and Hydrolysis General Procedure followed by repurification via 30 reverse phase préparative chromatography (C18, 0.1% TFA in water : MeCN) starting from the product from Step C, the TFA-salt of the desired product was obtained.
489
HRMS-ESI (m/z): [M+H]+ calcd for CjeHeFNsCbSz: 717.2805, found 717.2779.
Example 214:2-[[6-(l,3-Benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-methylamino]-5-[3-[4-[3-(dimethylamino)propyl]-2-fluoro-phenoxy]propyl]thiazole-4carboxylic acid
Step A: ethyl5-[3-[4-[3-(dimethylamino)propyl]-2-fluoro-phenoxy]propyl]-2-[methyl-[5methyl-6-[(E)-[3-(2-trimethylsilylethoxymethyl)-l,3-benzothiazol-2yliden eJamino]pyridazin -3 -yl]amino]thiazole-4-carboxylate
The mixture of 59 mg 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 éthanol was kept under 4 bar of hydrogen at rt for 18 h. After filtration through Celite, the product was concentrated to give 59 mg (99%) of the desired product.
*H NMR (500 MHz, DMSO-J6) 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); HRMS-ESI (m/z): [M+H]+ calcd for C39H53FN7O4S2S1: 794.3354 found: 794.3356.
Step B: 2-[[6-(l,3-benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-methyl-amino]-5-[3-[4[3-(diniethylamino)propyl]-2-fluoro-phenoxy]propyl]thiazole-4-carboxylic acid
To the 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 a 4 N solution of HCl (6.0 mmol) and was stirred at rt for 18 h. After the addition of 510 mg of 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% NH3) as eluents then by reverse phase préparative
490 chromatography (C18, 25 mM NH4HCO3 in water : MeCN) to give 3 mg (6%) of the desired product.
HRMS-ESI (m/z): [M+H]+ calcd for C31H35FN7O3S2: 636.2226 found: 636.2221.
Example 215:2-ir6-(1.3-Benzothiazol-2-vlamino)-5-methvl-pyridazin-3-vH-(4-hvdroxy-55 quinuclidin-l-ium-l-yl-pentyl)amino]-5-[3-[4-[3-(dimethylamino)prop-l-ynyl]-2-fluorophenoxy] propyl] thiazoIe-4-carboxylate
Step A: methyl 2-[[4-[tert-butyl(diphenyl)silyl]oxy-5-quinuclidin-l-ium-l-yl-pentyl]-[5methyl-6-[(Z)-[3-(2-trimethylsilylethoxymethyl)-l,3-benzothiazol-210 ylidene]amino]pyridazin-3-yl]amino]-5-[3-[4-[3-(dimethylamino)prop-l-ynyl]-2-fluorophenoxy]propyl]thiazole-4-carboxylate
Using Quaternary sait formation General Procedure starting from Préparation 5b and quinuclidine as the appropriate tertiary amine, the desired product was obtained.
Step B: 2-[[6-(l,3-benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-(4-hydroxy-515 quinuclidin-l-ium-l-yl-pentyl)amino]-5-[3-[4-[3-(dimethylamino)prop-l-ynyl]-2-fluorophenoxy]propyl]thiazole-4-carboxylate
Using Quaternary sait deprotection General Procedure starting from the product from Step A as the quaternary sait, the desired product was obtained.
HRMS-ESI (m/z): [M+2H]2+ calcd for C42H5iFN8O4S2: 407.1724, found 407.1724.
Example 216:2-i[5-(l-aza-4-azoniabicyclor2.2.2]octan-4-yl)-4-hydroxy-pentyl]-[6-(l,3benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]amino]-5-[3-[4-[3(dimethylamino)prop-l-ynyl]-2-fluoro-phenoxy]propyl]thiazole-4-carboxylate
491
Step A: methyl 2-[[5-(l-aza-4-azoniabicyclo[2.2.2]octan-4-yl)-4-[tertbutyl(diphenyl)silyl]oxy-pentyl]-[5-methyl-6-[(Z)-[3-(2-trimethylsilylethoxymethyl)-l,3benzothiazol-2-ylidene]amino]pyridazin-3-yl]amino]-5-[3-[4-[3-(dimethylamino)prop-l5 ynyl]-2-fluoro-phenoxy]propyl]thiazole-4-carboxylate
Using Quaternary sait formation General Procedure starting from Préparation 5b and 1,4diazabicyclo[2.2.2]octane as the appropriate tertiary amine, the desired product was obtained.
Step B: 2-[[5-(l -aza-4-azoniabicyclo[2.2.2]octan-4-yl) -4-hydroxy-pentyl]-[6-(l ,3benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]amino]-5-[3-[4-[3-(dimethylamino)prop10 1 -ynyl]-2-fluoro-phenoxy]propyl]thiazole-4-carboxylate
Using Quaternary sait deprotection General Procedure starting from the product from Step
A as the quaternary sait, the desired product was obtained.
HRMS-ESI (m/z): [M+2H]2+ calcd for C41H50FN9O4S2: 407.6700, found 407.6699.
Example 217:2-[[6-(l,3-BenzothiazoI-2-ylamino)-5-methyl-pyridazin-3-yl]-[4-hydroxy-515 (4-methylmorpholin-4-ium-4-yl)pentyl]amino]-5-[3-[4-[3-(dimethylamino)prop-l-ynyl]2-fIuoro-phenoxy]propyl]thiazole-4-carboxylate
492 b Step A: methyl2-[[4-[tert-butyl(diphenyl)silyl]oxy-5-(4-methylmorpholin-4-ium-4yl)pentyl]-[5-methyl-6-[(Z)-[3-(2-trimethylsilylethoxymethyl)-l,3-benzothiazol-2ylidene]amino]pyridazin-3-yl]amino]-5-[3-[4-[3-(dimethylamino)prop-l-ynyl]-2-fluorophenoxy]propyl]thiazole-4-carboxylate
Using Quaternary sait formation General Procedure starting from Préparation 5b and 4methylmorpholine as the appropriate tertiary amine, the desired product was obtained.
Step B: 2-[[6-(l,3-benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-[4-hydroxy-5-(4methylmorpholin-4-ium-4-yl)pentyl]amino]-5-[3-[4-[3-(dimethylamino)prop-l-ynyl]-2fluoro-phenoxy]propyl]thiazole-4-carboxylate
Using Quaternary sait deprotection General Procedure starting from the product from Step A as the quaternary sait, the desired product was obtained.
HRMS-ESI (m/z): [M+H]+ calcd for C40H48FN8O5S2: 803.3168, found 803.3165.
Example 218:3-[2-[3-[4-[3-[2-[[6-(l,3-Benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]methyl-amino]-4-carboxy-thiazol-5-yl]propoxy]-3-fluoro-phenyI]prop-215 ynylamino]ethyl-dimethyl-ammonio]propane-l-sulfonate
StepAy 3-[2-[tert-butoxycarbonyl-[3-[4-[3-[4-ethoxycarbonyl-2-[methyl-[5-methyl-6-[(Z)[3-(2-trimethylsilylethoxymethyl)-l,3-benzothiazol-2-ylidene]amino]pyridazin-3yl]amino]thiazol-5-yl]propoxy]-3-fluoro-phenyl]prop-2-ynyl]amino]ethyl-dimethyl20 ammonio]propane-l-sulfonate
159 mg of Préparation 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 when the conversion was reached >90%. The mixture was concentrated onto Celite and purified via flash column chromatography using EtOAc and MeOH (1.2% NH3) as
493
B eluents to give 126 mg (70%) of the desired product as yellow solid.
Hl NMR (500 MHz, DMSO-d6) 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 5 (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); 13C NMR (125 MHz, DMSO-de) 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; HRMS-ESI (m/z): [M+H]+ calcd for C49H68FN8O9S3S1: 1055.4019, found 1055.4026.
Step B: 3-[2-[3-[4-[3-[2-[[6-(l,3-benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-methylaniino]-4-carboxy-thiazol-5-yl]propoxy]-3-fluoro-phenyl]prop-2-ynylamino]ethyl-dimethylammonio]propane-l -sulfonate
Using Deprotection and Hydrolysis General Procedure followed by repurification via reverse phase préparative chromatography (C18, 25 mM NH4HCO3 in water : MeCN) starting 15 from the product from Step A, the TFA-salt of the desired product was obtained.
HRMS-ESI (m/z): [M+H]+ calcd for C36H42FN8O6S3: 797.2368, found 797.2343.
Example 219:2-i[6-(l,3-Benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-[5(trimethylammonio)pentyl] amino]-5-[3-[2-fluoro-4-[3-(methylamino)prop-lyny 1] phenoxy] propy 1] thiazole-4-carboxy late
Using Alkylation, Deprotection and Hydrolysis General procedure starting from Example 213, Step C and iodomethane, the TFA-salt of the desired product was obtained.
HRMS-ESI (m/z): [M+H]+ calcd for C37H44FN8O3S2: 731.2956, found 731.2967.
494
Example 220:2-[[6-(l,3-Benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-[4-hydroxy-5(l-methylpiperidin-l-ium-l-yl)pentyl]amino]-5-[3-[4-[3-(dimethylamino)prop-l-ynyl]-2fluoro-phenoxy]propyl]thiazole-4-carboxylate
Step A: methyl2-[[4-[tert-butyl(diphenyl)silyl]oxy-5-(l-methylpiperidin-l-ium-l-yl)pentyl][5-methyl-6-[(Z)-[3-(2-trimethylsilylethoxymethyl)-l,3-benzothiazol-2ylidene]amino]pyridazin-3-yl]amino]-5-[3-[4-[3-(dimethylamino)prop-l-ynyl]-2-fluorophenoxy]propyl]thiazole-4-carboxylate
Using Quaternary sait formation General Procedure starting from Préparation 5b and 110 methylpiperidine as the appropriate tertiary amine, the desired product was obtained.
StepB:2-[[6-(l,3-benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-[4-hydroxy-5-(lmethylpiperidin-l-ium-l-yl)pentyl]amino]-5-[3-[4-[3-(dimethylamino)prop-l-ynyl]-2fluoro-phenoxy]propyl]thiazole-4-carboxylate
Using Quaternary sait deprotection General Procedure starting from the product from Step 15 A as the quaternary sait, the desired product was obtained.
HRMS-ESI (m/z): [M+H]+ calcd for C41H50FN8O4S2: 801.3375, found 801.3378.
Example 221:2-iï6-(1.3-Benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-[5-(l,4dimethylpiperazin-l-ium-l-yl)-4-hydroxy-pentyl]amino]-5-[3-[4-[3(dimethylamino)prop-l-ynyl]-2-fluoro-phenoxy]propyl]thiazole-4-carboxylate
495
Step A: methyl 2-[[4-[tert-butyl(diphenyl)silyl]oxy-5-(l,4-dimethylpiperazin-l-ium-lyl)pentyl]-[5-methyl-6-[(Z)-[3-(2-trimethylsilylethoxymethyl)-l,3-benzothiazol-2ylidene]amino]pyridazin-3 -yl]amino]-5-[3-[4-[3-(dimethylamino)prop-l -ynyl]-2-fluoro5 phenoxy]propyl]thiazole-4-carboxylate
Using Quaternary sait formation General Procedure starting from Préparation 5b and 1,4dimethylpiperazine as the appropriate tertiary amine, the desired product was obtained.
Step B: 2-[[6-(l,3-benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-[5-(l,4dimethylpiperazin-l-ium-l-yl)-4-hydroxy-pentyl]amino]-5-[3-[4-[3-(dimethylamino)prop-l10 ynyl]-2-fluoro-phenoxy]propyl]thiazole-4-carboxylate
Using Quaternary sait deprotection General Procedure starting from the product of Step A as the quaternary sait, the desired product was obtained.
HRMS-ESI (m/z): [M+H]+ calcd for C41H51FN9O4S2: 816.3484, found 816.3489.
Example 222:2-[3-[4-[3-[2-[[6-(l,3-Benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]15 methyl-amino]-4-carboxy-thiazol-5-yl]propoxy]-3-fluoro-phenyl]prop-2ynylamino]ethyl-trimethyl-ammonium
496
Step A: 2-[tert-butoxycarbonyl-[3-[4-[3-[4-ethoxycarbonyl-2-[methyl-[5-methyl-6-[(Z)-[3(2-trimethylsilylethoxymethyl)-l,3-benzothiazol-2-ylidene]amino]pyridazin-3yl]amino]thiazol-5-yl]propoxy]-3-fluoro-phenyl]prop-2-ynyl]amino]ethyl-trimethylammonium
200 mg of Préparation 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 rt for 1.5 h. The reaction did not go further therefore stopped at this point, partially conversion was observed. The volatiles were evaporated and the crude mixture (contained Préparation 5h and the desired product) was transferred directly to the next step.
LC-MS-ESI (m/z): [M]+ calcd for C47H64FN8O6S2Si: 947.4, found 947.4.
Step B: 2-[3-[4-[3-[2-[[6-(l,3-benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-methylamino]-4-carboxy-thiazol-5-yl]propoxy]-3-fluoro-phenyl]prop-2-ynylamino]ethyl-trimethylammonium
Using Deprotection and Hydrolysis General Procedure followed by repurification via 15 reverse phase préparative chromatography (C18, 25 mM NH4HCO3 in water : MeCN) starting from the product from Step A, the desired product and Example 223 were separated and isolated.
HRMS-ESI (m/z): [M+H]+ calcd for C34H38FN8O3S2: 689.2487, found 689.2481.
Example 223:2-[[6-(l,3-Benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-methyl20 amino]-5-[3-[4-[3-[2-(dimethylamino)ethylamino]prop-l-ynyl]-2-fluorophenoxy]propyl]thiazole-4-carboxylic acid
Example 223 was isolated as side product from Example 222, Step B.
497
HRMS-ESI (m/z): [M+H]+ calcd for C33H36FN8O3S2: 675.2330, found 675.2324.
Example 224:2-[T6-(1.3-Benzothiazol-2-vlamino)-5-methvl-pyridazin-3-yl]-[6(trimethylammonio)hexyl]amino]-5-[3-[2-fluoro-4-[3-(methylamino)prop-lyny 1] phenoxy] propy 1] thiazole-4-carboxy late
Using Alkylation, Deprotection and Hydrolysis General procedure starting from Example 199, Step C and iodomethane, TFA-salt of the desired product was obtained.
‘El NMR (500 MHz, DMSO-t/6) 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), 10 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); 13C NMR (125 MHz, DMSO-î/6) 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; HRMS-ESI (m/z): [M+H]+ calcd for C38H44FN8O3S2: 745.3118, found 745.3110.
Example 225:2-[[6-(l,3-Benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-[5-[315 hydroxypropyl(dimethyl)ammonio]pentyl]amino]-5-[3-[4-[3-(dimethylamino)prop-lynyl]-2-fluoro-phenoxy]propyI]thiazole-4-carboxyIate
498
Step A : 5-fi5-i3-i4-f3-(dimethylamino)prop-l -ynyl]-2-fluoro-phenoxy]propyl]-4methoxycarbonyl-thiazol-2-yl]-[5-methyl-6-[(Z)-[3-(2-trimethylsilylethoxymethyl)-l,3benzothiazol-2-ylidene]amino]pyridazin-3-yl]amino]pentyl-(3-hydroxypropyl)-dimethyl- ammonium
Using Alkylation with in situ generated tosylate General Procedure followed by purification via reverse phase flash column chromatography (C18, 0.1% TFA in water : MeCN) starting from Préparation 5f and 3-(dimethylamino)propan-l-ol as the appropriate amine, the desired product was obtained.
1H NMR (500 MHz, DMSO-dô) 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); HRMS-ESI (m/z): [M]+ calcd for C47H66FNsO5S2Si: 933.4346, found 933.4342.
Step B: 2-[[6-(l,3-benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-[5-[3hydroxypropyl(dimethyl)ammonio]pentyl]amino]-5-[3-[4-[3-(dimethylamino)prop-l-ynyl]2-fluoro-phenoxy]propyl]thiazole-4-carboxylate
Using Deprotection and Hydrolysis General Procedure followed by repurification via 20 reverse phase préparative chromatography (C18, 0.1% TFA in water : MeCN) starting from the product from Step A, the TFA-salt of the desired product was obtained.
HRMS-ESI (m/z): [M+H]+ calcd for C40H50FN8O4S2: 789.3375, found 789.3376.
499
Example 226:2-[[6-(l,3-Benzothiazol-2-yIamino)-5-methyl-pyridazin-3-yl]-[5-[2hydroxyethyl(dimethyl)ammonio]pentyl]amîno]-5-[3-[4-[3-(dimethylamino)prop-lynyl]-2-fluoro-phenoxy]propyl]thiazole-4-carboxylate
Step A: 5-[[5-[3-[4-[3-(dimethylamino)prop-l-ynyl]-2-fluoro-phenoxy]propyl]-4methoxycarbonyl-thiazol-2-yl]-[5-methyl-6-[(Z) -[3-(2-trimethylsilylethoxymethyl) -1,3benzothiazol-2-ylidene]amino]pyridazin-3-yl]amino]pentyl-(2-hydroxyethyl)-dimethylammonium
Using Alkylation with in situ generated tosylate General Procedure followed by purification via reverse phase flash column chromatography (C18, 0.1% TFA in water : MeCN) starting from Préparation 5f and 2-(dimethylamino)éthanol as the appropriate amine, the desired product was obtained.
1H NMR (500 MHz, DMSO-d6) 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 (br„ 1H), 4.42 (t, 2H), 15 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); 13C NMR (125 MHz, DMSO-de) 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, 31.1, 26.7, 23.2, 23.2, 21.9, 17.9, 17.8, -0.9; HRMS-ESI (m/z): [M]+ calcd for
C46H64FN8O5S2Si: 919.4189, found 919.4192.
Step B: 2-[[6-(l,3-benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-[5-[2hydroxyethyl(dimethyl)ammonio]pentyl]amino]-5-[3-[4-[3-(dimethylamino)prop-l-ynyl]-2fluoro-phenoxy]propyl]thiazole-4-carboxylate
Using Deprotection and Hydrolysis General Procedure followed by repurification via
500 reverse phase préparative chromatography (C18, 0.1% TFA in water : MeCN) starting from the product from Step A, the TFA-salt of the desired product was obtained.
HRMS-ESI (m/z): [M+2H]2+ calcd for C39H49FN8O4S2: 388.16455, found 388.1639.
Example 227:2-IT6-(1.3-Benzothiazol-2-vlamino)-5-methyl-pyridazin-3-yl]-[55 [carboxymethyl(dimethyl)ammonio]pentyl]amino]-5-[3-[4-[3-(dimethylamino)prop-lynyl] -2-fluoro-phenoxy] propy 1] thiazole-4-carboxylate
Step A: 5-[[5-[3-[4-[3-(dimethylamino)prop-l-ynyl]-2-fluoro-phenoxy]propyl]-4methoxycarbonyl-thiazol-2-yl]-[5-methyl-6-[(Z)-[3-(2-trimethylsilylethoxymethyl)-l,310 benzothiazol-2-ylidene]amino]pyridazin-3-yl]amino]pentyl-(2-ethoxy-2-oxo-ethyl)dimethyl-ammonium
Using Alkylation with in situ generated tosylate General Procedure followed by purification via reverse phase flash column chromatography (C18, 0.1% TFA in water : MeCN) starting from Préparation 5f and ethyl 2-(dimethylamino) acetate as the appropriate amine, the 15 desired product was obtained.
Hl NMR (500 MHz, DMSO-d6) 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 20 (m, 2H), 1.37 (m, 2H), 1.22 (t, 3H), 0.92 (m, 2H), -0.10(s, 9H); HRMS-ESI (m/z): [M]+ calcd for C48H66FN8O6S2Si: 961.4294, found 961.4293.
Step B: 2-[[6-(l,3-benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-[5[carboxymethyl(dimethyl)ammonio]pentyl]amino]-5-[3-[4-[3-(dimethylamino)prop-l -ynyl]20487
501
2-fluoro-phenoxy]propyl]thiazole-4-carboxylate
Using Deprotection and Hydrolysis General Procedure followed by repurification via reverse phase préparative chromatography (C18, 0.1% TFA in water : MeCN) starting from the product from Step A, the TFA-salt of the desired product was obtained.
HRMS-ESI (m/z): [M+H]+ calcd for C39H46FN8O5S2: 789.3011, found 789.2995.
Example 228:3-il5-ri6-(l,3-Benzothiazol-2-ylamino)-5-methyl-p.yridazin-3-yl]-[4carboxy-5-[3-[2-fluoro-4-[(Ê')-3-oxoprop-l-enyI]phenoxy]propyl]thiazol-2-yl]amino]-2hydroxy-pentyl]-dimethyl-ammonio]propane-l-sulfonate
Example 228 was isolated as side-product from the synthesis of Example-146.
HRMS-ESI (m/z): [M+H]+ calcd for C38H45FN7O8S3: 842.2470, found 842.2494.
Example 229:2-[[6-(l,3-Benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-[5-[2hydroxyethyl(dimethyl)ammonio]pentyl]amino]-5-[3-[2-fluoro-4-[3-(methylamino)propl-ynyl]phenoxy]propyl]thiazole-4-carboxylate
502
Step A: 5-[[5-[3-[4-[3-[tert-butoxycarbonyl(methyl)amino]prop-l-ynyl]-2-fluorophenoxy]propyl]-4-methoxycarbonyl-thiazol-2-yl]-[5-methyl-6-[(Z)-[3-(2trimethylsilylethoxymethyl)-l,3-benzothiazol-2-ylidene]amino]pyridazin-3-yl]amino]pentyl(2-hydroxyethyl) -dimethyl-ammonium
Using Alkylation with in situ generated tosylate General Procedure followed by purification via reverse phase flash column chromatography (C18, 0.1% TFA in water : MeCN) starting from Préparation 5e and 2-(dimethylamino)ethanol as the appropriate amine, the desired product was obtained.
‘H NMR (500 MHz, DMSO-dô) 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); HRMS-ESI (m/z): [M]+ calcd for CsoHyoFNsOvSzSi: 1005.4557, found 1005.4554.
StepB:2-[[6-(l,3-benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-[5-[2hydroxyethyl(dimethyl)ammonio]pentyl]amino]-5-[3-[2-fluoro-4-[3-(methylamino)prop-lynyl]phenoxy]propyl]thiazole-4-carboxylate
Using Deprotection and Hydrolysis General Procedure followed by repurification via reverse phase préparative chromatography (C18, 0.1% TFA in water : MeCN) starting from the product from Step A, the TFA-salt of the desired product was obtained.
HRMS-ESI (m/z): [M+H]+ calcd for C38H46FN8O4S2: 761.3062, found 761.3057.
Example 230:2-[[6-(l,3-Benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-[5-[3hydroxypropyI(dimethyl)ammonio]pentyl]amino]-5-[3-[2-fluoro-4-[3(methy lamino) prop- 1-yny 1] phenoxy] propyl] thiazole-4-carboxylate
503
Step A: 5-[[5-[3-[4-[3-[tert-butoxycarbonyl(methyl)amino]prop-l-ynyl]-2-fluorophenoxy]propyl]-4-methoxycarbonyl-thiazol-2-yl]-[5-methyl-6-[(Z)-[3-(2trimethylsilylethoxymethyl)-l,3-benzothiazol-2-ylidene]amino]pyridazin-3-yl]amino]pentyl5 (3-hydroxypropyl) -dimethyl-ammonium
Using Alkylation with in situ generated tosylate General Procedure followed by purification via reverse phase flash column chromatography (C18, 0.1% TFA in water : MeCN) starting from Préparation 5e and 3-(dimethylamino)propan-l-ol as the appropriate amine, the desired product was obtained.
XH NMR (500 MHz, DMSO-cfo) 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), -0.11 (s, 9H); HRMS-ESI (m/z): [M]+ calcd for C5iH72FN8O7S2Si: 1019.4713, found 1019.4713.
Step B: 2-[[6-(l,3-Benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-[5-[3hydroxypropyl(dimethyl)ammonio]pentyl]amino]-5-[3-[2-fluoro-4-[3-(methylamino)prop-lynyl]phenoxy]propyl]thiazole-4-carboxylate
Using Deprotection and Hydrolysis General Procedure followed by repurification via 20 reverse phase préparative chromatography (C18, 0.1% TFA in water : MeCN) starting from the product from Step A, the TFA-salt of the desired product was obtained.
HRMS-ESI (m/z): [M+H]+ calcd for C39H48FN8O4S2: 775.3218, found 775.3213.
504
Example 23l:2-[r6-(l.3-Benzothiazol-2-vlamino)-5-methvl-pyridazin-3-yl1-l5[carboxy methyl (dimethyl) ammonio] pentyl] amino] -5- [3- [2-fluoro-4- [3(methylamino)prop-l-ynyl]phenoxy]propyI]thiazole-4-carboxylate
Step A: 5-[[5-[3-[4-[3-[tert-butoxycarbonyl(methyl)amino]prop-1-ynyl]-2-fluorophenoxy]propyl]-4-methoxycarbonyl-thiazol-2-yl]-[5-methyl-6-[(Z)-[3-(2trimethylsilylethoxymethyl)-l,3-benzothiazol-2-ylidene]amino]pyridazin-3-yl]amino]pentyl(2-ethoxy-2-oxo-ethyl)-dimethyl-ammonium
Using Alkylation with in situ generated tosylate General Procedure followed by purification via reverse phase flash column chromatography (C18,0.1% TFA in water : MeCN) starting from Préparation 5e and ethyl 2-(dimethylamino)acetate as the appropriate amine, the desired product was obtained.
Hl NMR (500 MHz, DMSO-·) 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-ESI (m/z): [M]+ calcd for C52H72FN8O8S2Si: 1047.4662, found 1047.468.
Step B: 2-[[6-(l,3-benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-[520 [carboxymethyl(dimethyl)ammonio]pentyl]amino]-5-[3-[2-fluoro-4-[3-(methylamino)prop-
-ynyl]phenoxy]propyl]thiazole-4-carboxylate
Using Deprotection and Hydrolysis General Procedure followed by repurification via reverse phase préparative chromatography (C18, 0.1% TFA in water : MeCN) starting from the product from Step A, the TFA-salt of the desired product was obtained.
HRMS-ESI (m/z): [M+H]+ calcd for C38H44FN8O5S2: 775.2855, found 775.2857.
505
Example 232:2-[[6-(l,3-Benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-[4(trimethyIammonio)butyl]amino]-5-[3-[2-fluoro-4-[3-(methylamino)prop-lyny 1] phenoxy] propyl] thiazole-4-carboxylate
Step A: 4-[[5-[3-[4-[3-[tert-butoxycarbonyl(methyl)amino]prop-l-ynyl]-2-fluorophenoxy]propyl]-4-methoxycarbonyl-thiazol-2-yl]-[5-methyl-6-[(Z)-[3-(2trimethylsilylethoxymethyl)-l,3-benzothiazol-2-ylidene]amino]pyridazin-3-yl]amino]butyltrimethyl-ammonium
Using Alkylation with in situ generated tosylate General Procedure starting from Préparation 5c and Ν,Ν-dimethylmethanamine as the appropriate amine, the TFA-salt of the desired product was obtained.
*H NMR (500 MHz, DMSO-d6) 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);13C NMR (125 MHz, DMSO-d6) 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, 24.4, 23.3,19.8,18.0,17.8, -1.0; HRMS-ESI (m/z): [M]+ calcd for C48H66FN8O6S2Si: 961.4294, found 961.4312.
Step B:2-ii6-(l,3-benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-[4(trimethylammonio)butyl]amino]-5-[3-[2-fluoro-4-[3-(methylamino)prop-lynyl]phenoxy]propyl]thiazole-4-carboxylate
Using Deprotection and Hydrolysis General Procedure followed by repurification via reverse phase préparative chromatography (C18, 0.1% TFA in water : MeCN), starting from the product from Step A, the TFA-salt of the desired product was obtained.
506 • HRMS-ESI (m/z): [M]+ calcd for C36H42FN8O3S2: 717.2800, found 717.2800.
Example 233:2-[[6-(l,3-Benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-methylamino]-5-[3-[4-[3-(dimethylamino)prop-l-ynyl]phenoxy]propyl]thiazole-4-carboxylic acid
Step A: ethyl5-[3-[4-[3-(dimethylamino)prop-l-ynyl]phenoxy]propyl]-2-[methyl-[5-methyl6-[(Z)-[3-(2-trimethylsilylethoxymethyl)-l,3-benzothiazol-2-ylidene]amino]pyridazin-3yl]amino]thiazole-4-carboxylate
Using Alkylation of Silyl-Protected Phénols General Procedure starting from 219 mg of 10 Préparation 5g (0.30 mmol, 1 eq.) and 100 mg of Préparation 6j (0.30 mmol, 1 eq.) followed by purification via flash column chromatography using EtOAc and MeOH (containing 1.2% NH3) as eluents to give 37 mg (12%) of the desired product as dark yellow sticky compound.
Ή NMR (500 MHz, DMSO-dô) 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, 15 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); HRMS-ESI (m/z): [M+H]+ calcd for C39H5oN704S2Si: 772.3129, found 772.3119.
Step B: 2-[[6-(l,3-benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-methyl-amino]-5-[3-[4[3-(dimethylamino)prop-l-ynyl]phenoxy]propyl]thiazole-4-carboxylic acid
Using Deprotection and Hydrolysis General Procedure followed by repurification via reverse phase préparative chromatography (C18, 0.1% TFA in water : MeCN) starting from the product from Step A, the TFA-salt of the desired product was obtained.
HRMS-ESI (m/z): [M+H]+ calcd for C31H32N7O3S2: 614.2002, found 614.1998.
507
Example234:2-[[6-(l,3-Benzothiazol-2-ylamino)-5-methyI-pyridazin-3-yl]-methylamino]-5-[3-[4-[l-[(dimethylamino)methyl]-3bicydoU.7J]pentanyl]phenoxy]propyl]thiazole-4-carboxylic acid
Step A: ethyl5-[3-[4-[l-[(dimethylamino)methyl]-3bicyclo[l .1.1 ]pentanyl]phenoxy]propyl]-2-[methyl-[5-methyl-6-[ (Z) -[3-(2trimethylsilylethoxymethyl)-l,3-benzothiazol-2-ylidene]amino]pyridazin-3yl]amino]thiazole-4-carboxylate
Using Alkylation General Procedure starting from 47 mg of Préparation 6i (0.22 mmol, 1 eq.) and 157 mg of Préparation 5g (0.22 mmol, 1 eq.) followed by purification via flash column chromatography using EtOAc and MeOH (containing 1.2% NH3) as eluents to give 130 mg (74%) of the desired product as yellow solid.
Hl NMR (500 MHz, DMSO-d6) 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, 9H); HRMS-ESI (m/z): [M+H]+ calcd for C42H56N7O4S2S1: 814.3599, found 814.3592.
Step B: 2-[[6-(l,3-benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-methyl-amino]-5-[3-[4[l-[(dimethylamino)methyl]-3-bicyclo[l.l.l]pentanyl]phenoxy]propyl]thiazole-4-carboxylic acid
Using Deprotection and Hydrolysis General Procedure followed by repurification via reverse phase préparative chromatography (C18, 0.1% TFA in water : MeCN) starting from the product from Step A, the TFA-salt of the desired product was obtained.
HRMS-ESI (m/z): [M+H]+ calcd for C34H38N7O3S2: 656.2472, found 656.2467.
508
Example 235:3-[3-[[6-(l,3-Benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-[4carboxy-5-[3-[2-fluoro-4-[3-(methylamino)prop-l-ynyl]phenoxy]propyl]thiazol-2yl]amino]propyl-dimethyl-ammonio]propane-l-sulfonate
Using Alkylation, Deprotection and Hydrolysis General procedure starting from Example 200, Step C and oxathiolane 2,2-dioxide. After alkylation, the sulfonated intermediate was isolated (HRMS-ESI (m/z): [M+H]+ calcd for C49H68FN8O9S3Si: 1055.4025, found 1055.4028) then reacted with HFxPyridine (HRMS-ESI (m/z): [M+H]+ calcd for Css^sFNsOôSs: 825.2686, found 825.2675) and finally hydrolyzed with LiOHxH2O. After purification it resulted in the TFA-salt of the desired product.
HRMS-ESI (m/z): [M+H]+ calcd for C37H44FN8O6S3: 811.2530, found 811.2525.
Example 236: 2-({6-[(l,3-Benzothiazol-2-yl)amino]-5-methylpyridazin-3yl}(methyl)amino)-5-[3-(2-fluoro-4-{3-[methyl({4-[3-(trifluoromethyl)-3H-diazirin-3yl]phenyl}methyl)amino]prop-l-yn-l-yl}phenoxy)propyl]-l,3-thiazole-4-carboxylic acid
509
Step A : ethyl 5-[3-(2-fluoro-4-{3-[methyl({4-[3-(trifluoromethyl)-3H-diazirin-3yl]phenyl}methyl)amino]prop-l-yn-l-yl}phenoxy)propyl]-2-[methyl(5-methyl-6-{[(2Z)-3{[2-(trimethylsilyl)ethoxy]niethyl}-2,3-dihydro-l,3-benzothiazol-2-ylidene]amino}pyridazin3-yl)amino]-l,3-thiazole-4-carboxylate
To a solution of the product from Préparation 5j (20 mg, 25.8 pmol, 1 eq) in dry dimethylformamide (1 mL) was added potassium carbonate (3.9 mg, 28.4 pmol, 1.1 eq) followed by 3-[4-(bromomethyl)phenyl]-3-(trifluoromethyl)diazirine (7.9 mg, 28.4 pmol, 1.1 eq) and the mixture was stirred for 16 h at ambient température. The reaction was diluted with 5% aqueous ammonium hydroxide (10 mL), extracted with ethyl acetate (2 x 10 mL), and 10 the combined organic extracts were successively washed with 5% aqueous ammonium hydroxifluorade (10 mL) and brine (10 mL), dried (magnésium sulfate), and concentrated in vacuo. Purification by automated flash column chromatography (CombiFlash Rf, 4 g RediSep™ silica cartridge) eluting with a gradient of 0 - 50% ethyl acetate in Ao-heptane afforded the desired product as a white solid (27.4 mg) that was used directly in the next step 15 without further characterisation.
Step B: ethyl 2-({6-[(l,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)5-[3-(2-fluoro-4-{3-[methyl({4-[3-(trifluoromethyl)-3H-diazirin-3yl]phenyl}methyl)amino]prop-l-yn-l-yl}phenoxy)propyl]-l,3-thiazole-4-carboxylate
To a solution of the product from Step A (27.4 mg, 28 pmol, 1 eq) in acetonitrile (1 mL) was 20 added pyridinium poly(hydrogen fluoride) (101 pL, 0.78 mmol, 28 eq) and the mixture was heated at 60 °C for 20 h. The reaction was diluted with 5% aqueous ammonium hydroxide (10 mL), extracted with ethyl acetate (2 x 10 mL), and the combined organic extracts were dried (magnésium sulfate) and concentrated in vacuo. Purification by reverse phase automated flash chromatography (CombiFlash Rf, C18 5.5g RediSep aq Gold column) eluting with a gradient 25 of 5 - 95% acetonitrile in water afforded the desired product as a yellow solid (6.7 mg, 0.01 mmol, 28%).
LC/MS (C41H37F4N9O3S2) 844 [M+H]+; RT 1.64 (LCMS-V-B1)
Step C: 2-({6-[(l,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)-5-[3(2-fluoro-4-{3-[methyl({4-[3-(trifluoromethyl)-3H-diazirin-3-yl]phenyl}methyl)amino]prop30 l-yn-l-yl}phenoxy)propyl]-l,3-thiazole-4-carboxylic acid
To a solution of the product from Step B (6.7 mg, 0.01 mmol, 1 eq) in 1:1 tetrahydrofuran / methanol (0.5 mL) was added 2M aqueous sodium hydroxide (79.4 pL, 0.16 mmol, 20 eq) and
510 φ the mixture was stirred at ambient température for 4 h then concentrated in vacuo. The residue was diluted with water (0.2 mL) and adjusted to pH 5 with 2M aqueous hydrochloric aced. Purification by reverse phase automated flash chromatography (CombiFlash Rf, C18 5.5g RediSep aq Gold column) eluting with a gradient of 5 - 95% acetonitrile in water afforded the 5 desired product as an orange / brown solid (3.5 mg, 4.29 pmol, 54%).
LC/MS (C39H33F4N9O3S2) 816 [M+H]+; RT 1.53 (LCMS-V-B1)
Example 237:2-[[6-(l,3-Benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-(3phosphonopropyl)amino]-5-[3-[2-fluoro-4-[3-(methyIamino)prop-lyny 1] phenoxy] propy 1] thiazole-4-carboxylic acid
Step A: methyl 2-[tert-butoxycarbonyl(3-diethoxyphosphorylpropyl)amino]-5-[3-[4-[3-[tertbutoxycarbonyl(methyl)amino]prop-l-ynyl]-2-fluoro-phenoxy]propyl]thiazole-4carboxylate
Using Alkylation General Procedure starting from 500 mg Préparation 1b (0.87 mmol, 1 15 eq.) and 336 mg of l-bromo-3-diethoxyphosphoryl-propane (1.30 mmol, 1.5 eq.) as the appropriate halide, 517 mg (79%) of the desired product was obtained.
JH NMR (500 MHz, DMSO-ifc) δ ppm 7.30 (dd, 1H), 7.21 (dm, 1 H), 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); HRMS-ESI (m/z): [M+H]+ 20 calcd for C35H52FN3O10PS: 756.3090, found 756.3095.
Step B: methyl 5-[3-[4-[3-[tert-butoxycarbonyl(methyl)amino]prop-l-ynyl]-2-fluorophenoxy]propyl]-2-(3-diethoxyphosphorylpropylamino)thiazole-4-carboxylate
Using Deprotection with HFIP General Procedure starting from 512 mg of the product from
511
Step A (0.68 mmol, 1 eq.), 210 mg (47%) of the desired product was obtained.
XH NMR (500 MHz, DMSO-rfc) 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); X3C NMR (125 MHz, 5 DMSO-îZô) 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;
HRMS-ESI (m/z): [M+H]+ calcd for C30H44FN3O8PS: 656.2565, found 656.2555.
Step C: methyl 5-[3-[4-[3-[tert-butoxycarbonyl(methyl)amino]prop-l-ynyl]-2-fluorophenoxy]propyl]-2-[3-diethoxyphosphorylpropyl-[5-methyl-6-[(Z)-[3-(2trimethylsilylethoxymethyl)-l,3-benzothiazol-2-ylidene]amino]pyridazin-3- yl]amino]thiazole-4-carboxylate
Using Buchwald General Procedure II starting from 450 mg of the product from Step B (0.69 mmol, 1 eq.) and 363 mg of Préparation 4a (0.89 mmol, 1.3 eq.) as the appropriate halide, 540 mg (77%) of the desired product was obtained.
HRMS-ESI (m/z): [M+H]+ calcd for C4sH66FN7O9PS2Si: 1026.3849, found 1026.3849.
StepD:2-[f6-(1.3-benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-(3phosphonopropyl)amino]-5-[3-[2-fluoro-4-[3-(methylamino)prop-lynyl]phenoxy]propyl]thiazole-4-carboxylic acid
Using Deprotection and hydrolysis of phosphonic acid dérivatives General Procedure followed by repurification via reverse phase préparative chromatography (C18, 0.1% TFA in 20 water : MeCN) starting from the product from Step C, the TFA-salt of the desired product was obtained.
HRMS-ESI (m/z): [M+H]+ calcd for C32H34FN7O6PS2: 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-(dimethyIamino)prop-l-yn-l-yl]-225 fluorophenoxy}propyl)-l,3-thiazole-4-carboxylic acid
512
Step A: methyl 2-[tert-butoxycarbonyl-(4-ethoxy-4-oxo-butyl)amino]-5-[3-[4-[3(dimethylamino)prop-l-ynyl]-2-fluoro-phenoxy]propyl]thiazole-4-carboxylate
Using Alkylation General Procedure starting from Préparation le and ethyl 45 bromobutanoate, 0.48 g (77%) of the desired product was obtained.
XH 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); X3C 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; HRMS-ESI 10 (m/z): [M+H]+ calcd for C30H41FN3O7S: 606.2649, found 606.2640.
Step B: methyl 5-[3-[4-[3-(dimethylamino)prop-l-ynyl]-2-fluoro-phenoxy]propyl]-2-[(4ethoxy-4-oxo-butyl)amino]thiazole-4-carboxylate
Using Deprotection with HFIP General Procedure starting from the product from Step A, 0.32 g (83%) of the desired product was produced.
XH NMR (400 MHz, DMSO-d6) 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); X3C NMR (100 MHz, DMSO-î/6) 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; HRMS-ESI (m/z): [M+H]+ calcd for C25H33FN3O5S: 506.2125, found 506.2114.
Step C: methyl 5-[3-[4-[3-(dimethylamino)prop-l-ynyl]-2-fluoro-phenoxy]propyl]-2-[(4ethoxy-4-oxo-butyl)-[5-methyl-6-[(Z)-[3-(2-trimethylsilylethoxymethyl)-l,3-benzothiazol-2ylidene]amino]pyridazin-3-yl]amino]thiazole-4-carboxylate
Using Buchwald General Procedure II starting from the product from Step B, and Préparation 4a, 0.26 g (48%) of the desired product was produced.
XH NMR (500 MHz, DMSO-t/6) 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,
513 φ 2Η), 4.02 (q, 2Η), 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); HRMSESI (m/z): [M+H]+ calcd for C43H55FN7O6S2Si: 876.3409, found 876.3403; HRMS-ESI (m/z): [M+H]+ calcd for C42H53FN7O6S2Si (double Me-ester): 862.3252, found 862.3237.
step D: 2-({6-[ (l,3-Benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(3carboxypropyl)amino)-5-(3-{4-[3-(dimethylamino)prop-l-yn-l-yl]-2fluorophenoxy}propyl)-l,3-thiazole-4-carboxylic acid
Using Deprotection and Hydrolysis General Procedure followed by repurification via reverse phase préparative chromatography (C18, 0.1% TFA in water : MeCN) starting from the 10 product from Step C, TFA-salt of the desired product was obtained.
HRMS-ESI (m/z): [M+H]+ calcd for C34H35FN7O5S2: 704.2125, found 704.2116.
Example 239:2-({6-[(l,3-Benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(3phosphonopropyl)amino)-5-(3-{4-[3-(dimethylamino)prop-l-yn-l-yl]-2fluorophenoxy}propyl)-l,3-thiazole-4-carboxylic acid
Step A: methyl 2-[tert-butoxycarbonyl(3-diethoxyphosphorylpropyl)amino]-5-[3-[4-[3(dimethylamino)prop-l-ynyl]-2-fluoro-phenoxy]propyl]thiazole-4-carboxylate
Using Alkylation General Procedure starting from Préparation le and l-bromo-3diethoxyphosphoryl-propane as the appropriate halide, 0.52 g (77%) ofthe desired product was obtained.
iH NMR (500 MHz, DMSO-d6) δ ppm 7.29 (dd, 1H), 7.20 (dm, 1 H), 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); 13C NMR (125 MHz, DMSO-î/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,
514 φ 16.7; HRMS-ESI (m/z): [M+H]+ calcd for C31H46FN3O8PS: 670.2727, found 670.2716.
Step B: methyl 2-(3-diethoxyphosphorylpropylamino)-5-[3-[4-[3-(dimethylamino)prop-lynyl]-2-fluoro-phenoxy]propyl]thiazole-4-carboxylate
Using Deprotection with HFIP General Procedure starting from the product from Step A,
0.37 g (86%) of the desired product was obtained.
Ή NMR (500 MHz, DMSO-ri6) 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); HRMS-ESI (m/z): [M+H]+ calcd for C26H38FN3O6PS: 570.2203, found 570.2183.
Step C: methyl 2-[3-diethoxyphosphorylpropyl-[5-methyl-6-[(Z)-[3-(2trimethylsilylethoxymethyl)-l,3-benzothiazol-2-ylidene]amino]pyridazin-3-yl]amino]-5-[3[4-[3-(dimethylamino)prop-l-ynyl]-2-fluoro-phenoxy]propyl]thiazole-4-carboxylate
Using Buchwald General Procedure II starting from the product from Step B and Préparation 4a as the appropriate halide, 0.51 g (87%) of the desired product was obtained.
15 XH NMR (500 MHz, DMSO-î/6) 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); 13C NMR (125 MHz, DMSO-d6) δ ppm 163.1, 128.9, 127.2, 123.5, 123.2, 119.2, 117.4, 115.5, 20 112.0, 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, - .0; HRMS-ESI (m/z): [M+H]+ calcd for C44H6oFN707PS2Si: 940.3487, found 940.3471.
Step D: 2-({6-[(l,3-Benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(3phosphonopropyl)amino)-5-(3-{4-[3-(dimethylamino)prop-l-yn-l-yl]-2fluorophenoxy}propyl)-l,3-thiazole-4-carboxylic acid
Using Deprotection and hydrolysis of phosphonic acid dérivatives General Procedure followed by repurification via reverse phase préparative chromatography (C18, 0.1% TFA in water : MeCN) starting from the product from Step C, TFA-salt of the desired product was obtained.
HRMS-ESI (m/z): [M+H]+ calcd for C33H36FN7O6PS2: 740.1890, found 740.1871.
515
Example 240:2-({6-[(l,3-Benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(3sulfopropyl)amino)-5-(3-{4-[3-(dimethylamino)prop-l-yn-l-yl]-2fluorophenoxy}propyl)-l,3-thiazole-4-carboxylic acid
Step A; 3-[tert-butoxycarbonyl-[5-[3-[4-[3-(dimethylamino)prop-l-ynyl]-2-fluorophenoxy]propyl]-4-methoxycarbonyl-thiazol-2-yl]amino]propane-l-sulfonic acid
Using Alkylation General Procedure starting from Préparation le and sodium 3bromopropane-l-sulfonate as the appropriate halide, 0.50 g (77%) of the desired product was obtained.
XH NMR (500 MHz, DMSO-z/e) δ ppm 7.33 (dd, 1 H), 7.23 (dm, 1 H), 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); 13C NMR (125 MHz, DMSO-î/6) 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; HRMS-ESI (m/z): [M+H]+ calcd for C27H37FN3O8S2: 614.2006, found 614.1993.
Step B: 3-[[5-[3-[4-[3-(dimethylamino)prop-l -ynyl]-2-fluoro-phenoxy]propyl]-4methoxycarbonyl-thiazol-2-yl]amino]propane-l -sulfonic acid
Using Deprotection with HFIP General Procedure starting from the product from Step A, 0.26 g (63%) of the desired product was obtained.
‘H NMR (500 MHz, DMSO-de) δ ppm 7.61 (t, 1H), 7.33 (dd, 1H), 7.24 (dd, 1H), 7.14 (t, 1H), 20 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); 13C NMR (125 MHz, DMSO-r/6) 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-ESI (m/z): [M+Na]+ calcd for C22H28FN3NaO6S2: 536.1301, found 536.1293.
Step C: 3-[[5-[3-[4-[3-(dimethylamino)prop-l -ynyl]-2-fluoro-phenoxy]propyl]-4
516 methoxycarbonyl-thiazol-2-yl]-[5-methyl-6-[(Z)-[3-(2-trimethylsilylethoxymethyl)-l,3benzothiazol-2-ylidene]amino]pyridazin-3-yl]amino]propane-l-sulfonic acid
Using Buchwald General Procedure II starting from the product from Step B and Préparation 4a as the appropriate halide, 0.27 g (66%) of the desired product was obtained.
XH NMR (500 MHz, DMSO-d6) 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); 13C NMR (125 MHz, DMSO-ifc) 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, 10 52.0, 48.7, 47.3, 46.2, 42.9, 30.9, 23.6, 23.1,17.9,17.8, -0.9; HRMS-ESI (m/z): [M+H]+ calcd for C40H51FN7O7S3S1: 884.2765, found 884.2757.
StepD^2-({6-[(l,3-Benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(3-sulfopropyl)amino)5-(3-{4-[3-(dimethylamino)prop-l-yn-l-yl]-2-fluorophenoxy}propyl)-l,3-thiazole-4carboxylic acid
Using Deprotection and Hydrolysis General Procedure followed by repurification via reverse phase préparative chromatography (C18, 0.1% TFA in water : MeCN) starting from the product from Step C, TFA-salt of the desired product was obtained.
HRMS-ESI (m/z): [M+H]+ calcd for C33H35FN7O6S3: 740.1795, found 740.1786.
Example 241:2-({6-[(l,3-Benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(420 sulfobutyl)amino)-5-(3-{4-[3-(dimethylamino)prop-l-yn-l-yl]-2-fluorophenoxy}propyl)l,3-thiazole-4-carboxylic acid
Step A: 4-[tert-butoxycarbonyl-[5-[3-[4-[3-(dimethylamino)prop-l-ynyl]-2-fluorophenoxy]propyl]-4-methoxycarbonyl-thiazol-2-yl]amino]butane-l-sulfonic acid
517
Using Alkylation General Procedure starting from oxathiane 2,2-dioxide and Préparation le, 0.29 g of the desired product was produced.
XH NMR (500 MHz, DMSO-d6) 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); 13C NMR (125 MHz, DMSO-d6) 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-ESI (m/z): [M+Na]+ calcd for C28H38FN3NaO8S2: 650.1982, found 650.1974.
Step B: 4-[[5-[3-[4-[3-(dimethylamino)prop-l-ynyl]-2-fluoro-phenoxy]propyl]-4methoxycarbonyl-thiazol-2-yl]amino]butane-l-sulfonic acid
Using Deprotection with HFIP General Procedure starting from the product from Step A, 0.12 g of the desired product was produced.
HRMS-ESI (m/z): [M+H]+ calcd for C23H31FN3O6S2: 528.1638, found: 528.1632.
Step C: 4-[[5-[3-[4-[3-(dimethylamino)prop-l-ynyl]-2-fluoro-phenoxy]propyl]-4methoxycarbonyl-thiazol-2-yl]-[5-methyl-6-[(Z)-[3-(2-trimethylsilylethoxymethyl)-l,3benzothiazol-2-ylidene]amino]pyridazin-3-yl]amino]butane-l-sulfonic acid
Using Buchwald General Procedure II starting from the product from Step B and Préparation 4a, 0.16 g of the desired product was produced.
*H NMR (500 MHz, DMSO-d6) 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); 13C NMR (125 MHz, DMSO-dô) ôppm 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, 22.9,17.8,17.8, -0.9; HRMS-ESI (m/z): [M+H]+ calcd for C4iH53FN7O7S3Si: 898.2922, found: 898.2907.
Step D: 2-({6-[(l,3-Benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(4-sulfobutyl)amino)5.(3f4-[3-(dimethylamino)prop-l-yn-l-yl]-2-fluorophenoxy}propyl)-l,3-thiazole-4carboxylic acid
Using Deprotection and Hydrolysis General Procedure starting from the product from Step C, 64 mg of the desired product was produced.
HRMS-ESI (m/z): [M+H]+ calcd for C34H37FN7O6S3: 754.1951, found: 754.1930.
518
Example 242:2-({6-[(l,3-Benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(4phosphonobutyl)amino)-5-(3-{4-[3-(dimethylamino)prop-l-yn-l-yl]-2fluorophenoxy}propyl) -l,3-thiazole-4-carboxylic acid
Step A: methyl 2-[tert-butoxycarbonyl(4-diethoxyphosphorylbutyl)amino]-5-[3-[4-[3(dimethylamino)prop-l-ynyl]-2-fluoro-phenoxy]propyl]thiazole-4-carboxylate
Using Alkylation General Procedure starting from l-bromo-4-diethoxyphosphoryl-butane and Préparation le, 1.28 g of the desired product was produced.
’H NMR (500 MHz, DMSO-dô) δ ppm 7.29 (dd, 1 H), 7.20 (dm, 1H), 7.13 (t, 1H), 4.10 (t, 2H), 10 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); 13C NMR (125 MHz, DMSO-dô) 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; HRMS-ESI (m/z): [M+H]+ calcd for C32H48FN3O8PS: 684.2884, found 684.2859.
Step B: methyl 2-(4-diethoxyphosphorylbutylamino)-5-[3-[4-[3-(dimethylamino)prop-lynyl]-2-fluoro-phenoxy]propyl]thiazole-4-carboxylate
Using Deprotection with HFIP General Procedure starting from the product from Step A, 0.92 g of the desired product was produced.
Hl NMR (500 MHz, DMSO-dô) δ ppm 7.61 (t, 1H), 7.32 (dd, 1H), 7.22 (dm, 1H), 7.13 (t, 1H), 20 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); 13C NMR (125 MHz, DMSO-dô) 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; HRMS-ESI (m/z): [M+H]+ calcd for C27H40FN3O6PS: 584.2359, found: 584.2340.
519
Step C: methyl 2-[4-diethoxyphosphorylbutyl-[5-methyl-6-[(Z)-[3-(2trimethylsilylethoxymethyl)-l,3-benzothiazol-2-ylidene]amino]pyridazin-3-yl]amino]-5-[3[4-[3-(dimethylamino)prop-l-ynyl]-2-fluoro-phenoxy]propyl]thiazole-4-carboxylate
Using Buchwald General Procedure II starting from the product from Step B and 5 Préparation 4a, 0.69 g of the desired product was produced.
1H NMR (500 MHz, DMSO-d6) 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 10 (m, 2H), -0.11 (s, 9H); HRMS-ESI (m/z): [M+H]+ calcd for C45H62FN7O7PS2Si: 954.3643, found: 954.3627.
StepDf2-({6-[(l,3-Benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(4phosphonobutyl)amino)-5-(3-{4-[3-(dimethylamino)prop-l-yn-l-yl]-2fluorophenoxy}propyl) -1,3-thiazole-4-carboxylic acid
Using Deprotection and hydrolysis of phosphonic acid dérivatives General Procedure starting from the product from Step C, 288 mg of the desired product was produced.
HRMS-ESI (m/z): [M+H]+ calcd for C34H38FN7O6PS2: 754.2047, found: 754.2036.
Example 243:2-({6-[(l,3-Benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(4carboxybutyl)amino)-5-(3-{4-[3-(dimethylamino)prop-l-yn-l-yl]-220 fluorophenoxy}propyl)-l,3-thiazole-4-carboxylic acid
Step A: methyl 2-[tert-butoxycarbonyl-(5-methoxy-5-oxo-pentyl)amino]-5-[3-[4-[3(dimethylamino)prop-l-ynyl]-2-fluoro-phenoxy]propyl]thiazole-4-carboxylate
Using Alkylation General Procedure starting from methyl 5-bromopentanoate and
520
Préparation le, 0.99 g of the desired product was produced.
Ή NMR (500 MHz, DMSO-d6) 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); 13C NMR (125 MHz, DMSO-d6) 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; HRMSESI (m/z): [M+H]+ calcd for C30H41FN3O7S: 606.2649, found 606.2644.
Step B: methyl 5-[3-[4-[3-(dimethylamino)prop-l-ynyl]-2-fluoro-phenoxy]propyl]-2-[(5methoxy-5-oxo-pentyl)amino]thiazole-4-carboxylate
Using Deprotection with HFIP General Procedure starting from the product from Step A, 0.72 g of the desired product was produced.
Ή NMR (500 MHz, DMSO-d6) 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); 13C NMR (125 MHz, DMSO-d6) 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; HRMS-ESI (m/z): [M+H]+ calcd for C25H33FN3O5S: 506.2125, found 506.2123.
Step C: methyl 5-[3-[4-[3-(dimethylamino)prop-l-ynyl]-2-fluoro-phenoxy]propyl]-2-[(5methoxy-5-oxo-pentyl)-[5-methyl-6-[(Z)-[3-(2-trimethylsilylethoxymethyl)-l,3-benzothiazol2-ylidene]amino]pyridazin-3-yl]amino]thiazole-4-carboxylate
Using Buchwald General Procedure II starting from the product from Step B and Préparation 4a, 0.79 g of the desired product was produced.
Ή NMR (500 MHz, DMSO-d6) 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, 2H), 0.91 (m, 2H), -0.11 (s, 9H); HRMSESI (m/z): [M+H]+ calcd for C43H55FN7O6S2S1: 876.3409, found 876.3378.
Step D: 2-({6-[(l,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(4carboxybutyl)amino) -5-(3-{4-[3-(dimethylamino)prop-l -yn-1 -yl]-2-fluorophenoxy}propyl) l,3-thiazole-4-carboxylic acid
Using Deprotection and Hydrolysis General Procedure starting from the product from Step C, 296 mg of the desired product was produced.
HRMS-ESI (m/z): [M+H]+ calcd for C35H37FN7O5S2: 718.2282, found: 718.2263.
521
Example 244:2-[(3-aminopropyl){6-[(l,3-benzothiazol-2-yl)amino]-5-methyIpyridazin-3yl}amino]-5-(3-{2-fluoro-4-[3-(methylammo)prop-l-yn-l-yl]phenoxy}propyl)-l,3thiazole-4-carboxylic acid
Example 245:2-r(3-azidopropvD{6-r(l,3-benzothiazoI-2-yl)amino]-5-methylpyridazin-3yl}amino]-5-(3-{2-fluoro-4-[3-(methylamino)prop-l-yn-l-yl]phenoxy}propyl)-l,3thiazole-4-carboxylic acid
Example 246:2-[{6-r(l,3-benzothiazol-2-yl)amino]-5-methylpyridazin-310 yl}(methyl)ammo]-5-[3-(4-{3-[(dimethylamino)methyl]bicyclo[l.l.l]pentan-l-yl}-2fluorophenoxy)propyl]-l,3-thiazole-4-carboxylic acid
522
Example 247:6-i{6-i(l,3-benzothiazoI-2-yl)amino]-5-methylpyridazin-3yI}(methyl)amino]-3-(l-{[3-(2-{[(3S)-3,4-dihydroxybutyl]amino}ethoxy)-5,7dimethyladamantan-l-yl]methyl}-5-methyl-lH-pyrazol-4-yl)pyridine-2-carboxylic acid
Using the Amine Substitution and Hydrolysis General procedure starting from Préparation 14 and 2-[(4S)-2,2-dimethyl-l,3-dioxolan-4-yl]ethanamine as the appropriate amine, the desired product was obtained.
HRMS-ESI (m/z): [M+H]+ calcd for C42H54N9O5S: 796.3969, found: 796.3965.
Example 248:6-R6-r(1.3-benzothiazol-2-yl)amino]-5-methylpyridazin-3yl}(methyl)amino]-3-(I-{[3-(2-{[(3Æ)-3,4-dihydroxybutyl]amino}ethoxy)-5,7dimethyladamantan-l-yl]methyl}-5-methyl-LELpyrazol-4-yl)pyridine-2-carboxylic acid
523
Example 249:6-R6-r(1.3-benzothiazol-2-yl)amino]-5-methylpyridazin-3yl}(methyl)amino]-3-[l-({3,5-dimethyl-7-[2-(methylamino)ethoxy]adamantan-lyl}methyl)-5-methyl-17/-pyrazoI-4-yl]pyridine-2-carboxyIic acid
Example 250:6-r{6-[(l,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3yl}(methyl)amino]-3-[l-({3-[2-(dimethylamino)ethoxy]-5,7-dimethyladamantan-lyl}methyl)-5-methyl-lH-pyrazol-4-yI]pyridine-2-carboxylic acid
524
Using the Amine Substitution and Hydrolysis General procedure starting from Préparation 14 and dimethylamine as the appropriate amine, the desired product was obtained.
HRMS-ESI (m/z): [M+H]+ calcd for C40H50N9O3S: 736.3757, found: 736.3751.
Example 25l:6-r{6-r(l.3-benzothiazol-2-yl)amino1-5-methylpyridazin-3yl}(methyl)amino]-3-{l-[(3,5-dimethyl-7-{2-[(2-sulfoethyl)amino]ethoxy}adamantan-lyl)melhyl|-5-methyl-l W-pyrazol-4-yl}pvridinc-2-carboxylic acid
Example 252:6- Γ16- r(l.3-benzothiazol-2-yl)amino1 -5-methylpyridazin-310 yl}(methyl)amino]-3-{l-[(3,5-dimethyl-7-{2-[methyl(2sulfoethyl)amino]ethoxy}adamantan-l-yl)methyl]-5-methyl-lH-pyrazol-4-yl}pyridine-2carboxylic acid
525
Example 253;2-R6-r(1.3-benzothiazol-2-yI)amino]-5-methylpyridazin-3-yl}(pent-4-yn-lyl)amino]-5-(3-{2-fluoro-4-[3-(methylamino)prop-l-yn-l-yl]phenoxy}propyl)-l,3thiazole-4-carboxyIic acid
Step A: methyl 2-[tert-butoxycarbonyl(5-trimethylsilylpent-4-ynyl)amino]-5-[3-(2-fluoro-4iodo-phenoxy)propyl]-l,3-thiazole-4-carboxylate
Using Mitsunobu General Procedure starting from Préparation la (10 mmol) as the appropriate carbamate and 5-trimethylsilylpent-4-yn-l-ol (2 eq.) as the appropriate alcohol, 6.0 10 g (89%) of the desired product was obtained.
XH NMR (500 MHz, DMSO-rf6) 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
526 φ (s, 2Η), 3.27 (t, 2H), 2.88 (t, 1H), 2.51 (s, 3H), 2.46 (s, 3H), 2.31 (m, 2H), 2.14 (m, 2H), 1.91 (m, 2H); HRMS-ESI (m/z): [M+H]+ calcd for C34H33FN7O3S2: 670.2070, found 670.2052.
Step B: methyl 5-[3-(2-fluoro-4-iodo-phenoxy)propyl]-2-(5-trimethylsilylpent-4-ynylamino)1,3-thiazole-4-carboxylate
Using Deprotection with HFIP General Procedure starting from 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 desired product was obtained.
XH NMR (500 MHz, DMSO-cfc) 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), 10 0.11 (s, 9H); 13C NMR (125 MHz, DMSO-d6) 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; HRMS-ESI (m/z): [M+H]+ calcd for C22H29FIN2O3SSÎ: 575.0697, found: 575.0695.
Step C: methyl 5-[3-[4-[3-[tert-butoxycarbonyl(methyl)amino]prop-l-ynyl]-2-fluorophenoxy]propyl]-2-(5-trimethylsilylpent-4-ynylamino)-l,3-thiazole-4-carboxylate
Using Sonogashira General Procedure starting from the product from Step B (4.46 g, 7.76 mmol, 1 eq.) and tert-butylN-methyl-N-prop-2-ynyl-carbamate (2.62 g, 15.53 mmol, 2 eq.) as the appropriate acetylene, 4.44 g (93%) of the desired product was obtained.
Ή NMR (500 MHz, DMSO-d6) 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), 20 2.00 (m, 2H), 1.68 (m, 2H), 1.41 (s, 9H), 0.11 (s, 9H); HRMS-ESI (m/z): [M+H]+ calcd for
C31H43FN3O5SS1: 616.2677, found: 616.2659.
StepD: methyl 5-[3-[4-[3-[tert-butoxycarbonyl(methyl)amino]prop-l-ynyl]-2-fluorophenoxy]propyl]-2-[[5-methyl-6-[(Z)-[3-(2-trimethylsilylethoxymethyl)-l,3-benzothiazol-2ylidene]amino]pyridazin-3-yl]-(5-trimethylsilylpent-4-ynyl)amino]-l,3-thiazole-425 carboxylate
Using Buchwald General Procedure I starting from the product from Step C (4.44 g, 7.2 mmol) as starting material in 1,4-dioxane at 120 °C, 3.85 g (54%) of the desired product was obtained.
‘H NMR (500 MHz, DMSO-d6) δ ppm 7.83 (d, 1H), 7.64 (s, 1H), 7.45 (dd, 1H), 7.42 (td, 1H), 30 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,
527 φ 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); 13C NMR (125 MHz, DMSO-ifc) 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; HRMS-ESI (m/z): [M+H]+ calcd for 5 C49H65FN7O6S2Si2: 986.3960, found: 986.3932.
Step E: 2-[[6-(l,3-benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-pent-4-ynyl-aminoJ-5[3-[2-fluoro-4-[3-(methylamino)prop-l-ynyl]phenoxy]propyl]-l,3-thiazole-4-carboxylic acid
Using Deprotection and Hydrolysis General Procedure starting from the product from Step D, followed by purification via reverse phase préparative chromatography (C18, 25 mM 10 NH4HCO3 in water : MeCN), the desired product was obtained.
HRMS-ESI (m/z): [M+H]+ calcd for C34H33FN7O3S2: 670.2070, found: 670.2052.
Example 254:2-r(4-aminobutvl){6-i(l,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3yl}amino]-5-(3-{4-[3-(dimethylamino)prop-l-yn-l-yl]-2-fluorophenoxy}propyl)-l,3thiazole-4-carboxylic acid
Example 255:2-l(4-azidobutvl){6-[(l,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3yl}amino]-5-(3-{4-[3-(dimethyIamino)prop-l-yn-l-yl]-2-fluorophenoxy}propyI)-l,3thiazoIe-4-carboxylic acid
528
Example 256:2-r{6-r(l.3-benzothiazol-2-yl)amino1-5-methylpyridazin-3-yl}(hex-5-yn-lyl)amino]-5-(3-{4-[3-(dimethylamino)prop-l-yn-l-yl]-2-fluorophenoxy}propyl)-l,3thiazole-4-carboxylic acid
Example 257:2-R6-r(l,3-benzothiazol-2-yl)amino]-5-methylpyridazm-3-yl}(pent-4-yn-lyl)ammo]-5-(3-{4-[3-(dimethylamino)prop-l-yn-l-yl]-2-fluorophenoxy}propyl)-l,3thiazole-4-carboxylic acid
529
Example 258:2-i(3-azidopropvl)-{6-r(1.3-benzothiazol-2-vl)aminol-5-methvlpyridazin-3yl}amino]-5-(3-{4-[3-(dimethylamino)prop-l-yn-l-yl]-2-fluorophenoxy}propyl)-l,3thiazoIe-4-carboxylic acid
Example 259:2-i(3-aminopropyl){6-[(l,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3yl}amino]-5-(3-{4-[3-(dimethylamino)prop-l-yn-l-yl]-2-fluorophenoxy}propyI)-l,3thiazole-4-carboxylic acid
Example 260:2-[(4-aminobutyl){6-[(l,3-benzothiazol-2-yI)ammo]-5-methylpyridazin-310 yI}amino]-5-(3-{2-fluoro-4-[3-(methylamino)prop-l-yn-l-yl]phenoxy}propyl)-l,3thiazole-4-carboxylic acid
530
Example 261:2-r(4-azidobutvl){6-[(l,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3yl}amino]-5-(3-{2-fluoro-4-[3-(methylamino)prop-l-yn-l-yI]phenoxy}propyl)-l,3thiazole-4-carboxylic acid
Example 262:2-ί{6-Γ(l,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(hex-5-yn-lyl)amino]-5-(3-{2-fluoro-4-[3-(methylammo)prop-l-yn-l-yl]phenoxy}propyl)-l,3thiazole-4-carboxylic acid
531
Example 263:2-r(4-aminobutvl){6-r(1.3-benzothiazol-2-vl)amino]-5-methvlpyridazin-3yl}ammo]-5-(3-{2-fluoro-4-[3-(methylammo)propyI]phenoxy}propyl)-l,3-thiazole-4carboxylic acid
Example 264:2-r(4-azidobutyl){6-F(l,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3yl}amino]-5-(3-{2-fluoro-4-[3-(methylamino)propyl]phenoxy}propyl)-l,3-thiazole-4carboxylic acid
Example 265:2-r{6-[(l,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(hex-5-yn-l10 yI)amino]-5-(3-{2-fluoro-4-[3-(methylamino)propyI]phenoxy}propyI)-l,3-thiazole-4carboxylic acid
532
Example 266: 5-[3-[4-[3-[ierAbutoxycarbonyl(methyl)amino]prop-l-ynyl]-2-fluorophenoxy]propyl]-2-[[5-methyl-6-[(Z)-[3-(2-trimethylsilylethoxymethyI)-l,3-benzothiazol2-ylidene]amîno]pyridazin-3-yl]-pent-4-ynyl-amino]thiazole-4-carboxylic acid
The mixture of 500 mg of the product from Example 253, Step D and 106 mg (2.5 mmol, 5 eq.) of LiOH*H2O in the mixture of 5 mL of THF and 2 mL of water was stirred at 40 °C for 16 h, then purified by crystallization from Et2O to give 356 mg (78%) of the desired product.
HRMS-ESI (m/z): [M+H]+ calcd for C45H55FN7O6S2Si: 900.3409, found: 900.3383
Example 267: 2-[5-azidopentyl-[6-[(l,3-benzothiazol-2-yl)amino]-5-methyl-pyridazin-3yl]amino]-5-[3-[2-fluoro-4-[3-(methylamino)prop-l-ynyl]phenoxy]propyl]-l,3-thiazole-4carboxylic acid
533
Step A: methyl 2-[tert-butoxycarbonyl-[5-[tert-butyl(dimethyl)silyl]oxypentyl]amino]-5-[3[4-[3-[tert-butoxycarbonyl(methyl)amino]prop-l-ynyl]-2-fluoro-phenoxy]propyl]-l,3thiazole-4-carboxylate
Using Mitsunobu General Procedure starting from Préparation 1b (0.7 mmol) as the appropriate carbamate and 5-[tert-butyl(dimethyl)silyl]oxypentan-l-ol (1.5 eq.) as the appropriate alcohol, 494 mg (91%) of the desired product was obtained.
LC/MS (C39H6oFN308SSiNa) 800 [M+Na]+.
Step B: methyl 5-[3-[4-[3-[tert-butoxycarbonyl(methyl)amino]prop-l-ynyl]-2-fluoro10 phenoxy]propyl]-2-[5-[tert-butyl(dimethyl)silyl]oxypentylamino]-l,3-thiazole-4-carboxylate
Using Deprotection with HFIP General Procedure starting from 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.
Ή NMR (500 MHz, DMSO4) δ ppm 7.57 (br., 1H), 7.31 (d, 1H), 7.21 (d, 1H), 7.12 (t, 1H), 15 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); 13C NMR (125 MHz, DMSO-ifo) 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; HRMS-ESI (m/z): [M+H]+ calcd for C34H53FN3O6SSi: 678.3408, found: 678.3393.
Step C: methyl 5-[3-[4-[3-[tert-butoxycarbonyl(methyl)amino]prop-l-ynyl]-2-fluorophenoxy]propyl]-2-[5-[tert-butyl(dimethyl)silyl]oxypentyl-[5-methyl-6-[(Z)-[3-(2trimethylsilylethoxymethyl)-l,3-benzothiazol-2-ylidene]amino]pyridazin-3-yl]amino]-l,3thiazole-4-carboxylate
534
Using Buchwald General Procedure I starting from the product from Step B (235 mg) as starting material in 1,4-dioxane at 120 °C, 220 mg (60%) of the desired product was obtained.
Ή NMR (500 MHz, DMSO-ifc) 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); 13C NMR (125 MHz, DMSO-d6) 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; HRMS-ESI (m/z): [M+H]+ calcd for C52H75FN7O7S2SÎ2: 1048.4692, found: 1048.4692.
Step D: methyl5-[3-[4-[3-[tert-butoxycarbonyl(methyl)amino]prop-l-ynyl]-2-fluorophenoxy]propyl]-2-[5-hydroxypentyl-[5-methyl-6-[(Z)-[3-(2-trimethylsilylethoxymethyl)-l,3benzothiazol-2-ylidene]amino]pyridazin-3-yl]amino]-l,3-thiazole-4-carboxylate
The mixture of the product from Step C (0.3 mmol) and 18 mg of camphor sulfonic acid (0.07 mmol, 0.25 eq.) in 2 mL of the 1:1 mixture of DCM and MeOH was stirred at 60 °C for 1.5 h. The product was purified by flash column chromatography using heptane and EtOAc as eluents to give 195 mg (66%) of the desired product.
>H NMR (500 MHz, DMSO-d6) 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); 13C NMR (125 MHz, DMSO-J6) 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-ESI (m/z): [M+Na]+ calcd for C46H6oFN7Na07S2SiNa: 956.3641, found: 956.3646.
Step E: methyl 2-[5-azidopentyl-[5-methyl-6-[(Z)-[3-(2-trimethylsilylethoxymethyl)-1,3benzothiazol-2-ylidene]amino]pyridazin-3-yl]amino]-5-[3-[4-[3-[tertbutoxycarbonyl(methyl)amino]prop-l-ynyl]-2-fluoro-phenoxy]propyl]-l,3-thiazole-4carboxylate
The mixture of 158 mg of the product from Step D, 70 pL of triethylamine (0.50 mmol, 3 eq.), and 83 mg of p-tolylsulfonyl 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
535 °C for 0.5 h. Product was purified by flash column chromatography using heptane and
EtOAc as eluents to give 120 mg (73%) of the desired product.
Ή NMR (500 MHz, DMSO-d6) 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, 2H), 1.40 (s, 9H), 0.92 (m, 2H), 0.11 (s, 9 H); HRMS-ESI (m/z): [M+H]+ calcd for C46H6oFNio06S2Si: 959.3892, found: 959.3892.
Step F: 2-[5-azidopentyl-[6-[(l,3-benzothiazol-2-yl)aminoJ-5-methyl-pyridazin-3-ylJaminoJ5-[3-[2-fluoro-4-[3-(methylamino)prop-l-ynyl]phenoxy]propyl]-l,3-thiazole-4-carboxylic acid
Using Deprotection and Hydrolysis General Procedure starting from the product from Step E, followed by purification via reverse phase préparative chromatography (C18, 0.1% TFA in water : MeCN), the desired product was obtained.
HRMS-ESI (m/z): [M+H]+ calcd for C34H36FN10O3S2: 715.2397, found: 715.2391.
Example 268: 6-[[6-(l,3-benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yI]-methylamino]-3-[l-[[3-[2-(3-hydroxypropylamino)ethoxy]-5,7-dimethyI-l-adamantyI]methyl]5-methyl-pyrazol-4-yl]pyridine-2-carboxylic acid
Ο. ΌΗ
Using the Amine Substitution and Hydrolysis General procedure starting from Préparation 14 and 3-aminopropane-l-ol as the appropriate amine, the desired product was obtained.
HRMS-ESI (m/z): [M+H]+ calcd for C41H52N9O4S: 766.3863, found: 766.3860.
536
Example 269: 6-[[6-(l,3-benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-methylamino]-3-[l-[[3-[2-(3-methoxypropylamino)ethoxy]-5,7-dimethyl-l-adamantyl]methyl]5-methyl-pyrazol-4-yl]pyridine-2-carboxylic acid
Using the Amine Substitution and Hydrolysis General procedure starting from Préparation 14 and 3-methoxypropan-l-amine as the appropriate amine, the desired product was obtained.
HRMS-ESI (m/z): [M+H]+ calcd for C42H54N9O4S: 780.4019, found: 780.4019.
Example 270: 6-[[6-(l,3-benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-methylamino]-3-[l-[[3,5-dimethyl-7-(2-morpholinoethoxy)-l-adamantyl]methyl]-5-methyl10 pyrazol-4-yl]pyridine-2-carboxylic acid
Using the Amine Substitution and Hydrolysis General procedure starting from Préparation 14 and morpholine as the appropriate amine, the desired product was obtained.
HRMS-ESI (m/z): [M+H]+ calcd for C42H52N9O4S: 778.3863, found: 778.3858.
537
Example 271: 6-[[6-(l,3-benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-methylamino]-3-[l-[[3-[2-(2-carboxyethylamino)ethoxy]-5,7-dimethyl-l-adamantyl]methyl]-5methyl-pyrazol-4-yl]pyridine-2-carboxylic acid
Using the Amine Substitution and Hydrolysis General procedure starting from Préparation
14, 3-aminopropanoic-acid as the appropriate amine, and 10 eq of K2CO3 as a base, the desired product was obtained.
HRMS-ESI (m/z): [M+H]+ calcd for C41H5QN9O5S: 780.3656, found: 780.3658.
Example 272: 6-[[6-(l,3-benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-methyl10 amino]-3-[l-[[3-[2-[(3-hydroxyphenyl)methylamino]ethoxy]-5,7-dimethyl-ladamantyl]methyl]-5-methyl-pyrazol-4-yl]pyridine-2-carboxylic acid
Using the Amine Substitution and Hydrolysis General procedure starting from Préparation 14 and 3-(aminomethyl)phenol as the appropriate amine, the desired product was obtained.
HRMS-ESI (m/z): [M+H]+ calcd for C45H52N9O4S: 814.3863, found: 814.3861.
538
The compounds of the following Examples 273-280 are synthesised using the Amine Substitution and Hydrolysis General procedure starting from the Préparation 14 and the appropriate amine.
Example 273: 6-[[6-(l,3-benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yI]-methyl5 amino]-3-[l-[[3,5-dimethyl-7-(2-pyrrolidin-l-ylethoxy)-l-adamantyl]methyl]-5-methylpyrazol-4-yl] pyridine-2-carboxy lie acid
Example 274: 6-[[6-(l,3-benzothiazoI-2-ylamino)-5-methyl-pyridazin-3-yl]-methylamino]-3-[l-[[3,5-dimethyl-7-[2-(4-methylpiperazin-l-yl)ethoxy]-l-adamantyl]methyl]-510 methyl-pyrazol-4-yl]pyridine-2-carboxylic acid
Example 275: 6-[[6-(l,3-benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-methylamino]-3-[l-[[3-[2-(4-hydroxybutylamino)ethoxy]-5,7-dimethyI-l-adamantyl]methyl]-5methyl-pyrazol-4-y 1] pyridine-2-carboxylic acid
539
Example 276: 6-[[6-(l,3-benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-methylamino]-3-[l-[[3,5-dimethyl-7-(2-piperazin-l-ylethoxy)-l-adamantyl]methyI]-5-methylpyrazol-4-yl]pyridine-2-carboxylic acid
Example 277: 6-[[6-(l,3-benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-methylamino]-3-[l-[[3-[2-[3-hydroxypropyl(methyl)amino]ethoxy]-5,7-dimethyl-ladamantyl]methyl]-5-methyl-pyrazol-4-yl]pyridine-2-carboxylic acid
540
Example 278: 6-[[6-(l,3-benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-methylamino]-3-[l-[[3-[2-[4-hydroxybutyl(methyl)amino]ethoxy]-5,7-dimethyl-ladamantyl]methyl]-5-methyl-pyrazol-4-yl]pyridine-2-carboxylic acid
Example 279: 6-[[6-(l,3-benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-methylamino]-3-[l-[[3-[2-[[3-hydroxy-2-(hydroxymethyl)propyl]amino]ethoxy]-5,7-dimethyl-ladamantyl]methyl]-5-methyl-pyrazol-4-yl]pyridine-2-carboxylic acid
Example 280: 6-[[6-(l,3-benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-methyl10 amino] -3- [1- [[3- [2- [bis(3-hydroxypropyl)amino]ethoxy] -5,7-dimethyl-ladamantyl]methyl]-5-methyl-pyrazol-4-yl]pyridine-2-carboxylic acid
541
Example 281: 2-[[6-(l,3-benzothiazol-2-ylamino)-5-methyI-pyridazin-3-yl]-methylamino]-5-[3-[2-fluoro-4-(3-piperazin-l-ylprop-l-ynyl)phenoxy]propyl]thiazole-4carboxylic acid
Example 282: 2-[[6-(l,3-benzothiazoI-2-yiamino)-5-methyl-pyridazin-3-yl]-methylamino]-5-[3-[2-fluoro-4-[3-(4-methyIpiperazin-l-yI)prop-lyny 1] phenoxy] propy 1] thiazole-4-carboxylic acid
542
Example 283: 2-[[6-(l,3-benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-methylamino]-5-[3-[2-fhioro-4-[3-[3-hydroxypropyl(methyl)amino]prop-lyny 1] phenoxy] propy 1] thiazole-4-carboxylic acid
Example 284; 2-[[6-(l,3-benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-methyIamino] -5- [3- [4- [3- [[(SSj-S^-dihydroxybutyl] -methyl-amino]prop-l-ynyl]-2-fluorophenoxy] propy 1] thiazole-4-carboxylie acid
Example 285: 2-[[6-(l,3-benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-methyl10 amino]-5-[3-[2-fluoro-4-[3-(3-hydroxypropylamino)prop-lynyl] phenoxy] propyl] thiazole-4-carboxylic acid
543
Example 286: 2-[[6-(l,3-benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-methylamino] -5-[3- [4-[3- [[(3S) -3,4-dihydroxybutyl]amino] prop-1-ynyl] -2-fluorophenoxy] propyl] thiazole-4-carboxylie acid
Example 287: 2-[[6-(l,3-benzothiazoI-2-ylamino)-5-methyl-pyridazin-3-yl]-methylamino]-5-[3-[2-fluoro-4-[3-[4-hydroxybutyl(methyl)amino]prop-lynyl]phenoxy]propyl]thiazole-4-carboxylic acid
544
Example 288: 2-[[6-(l,3-benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-methylamino]-5-[3-[4-[3-[[(3B)-3,4-dihydroxybutyl]-methyl-amino]prop-l-ynyl]-2-fluorophenoxy] propyl] thiazole-4-carboxylie acid
Example 289: 2-[[6-(l,3-benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-methylamino]-5-[3-[2-fluoro-4-[3-(4-hydroxybutylamino)prop-l-ynyl]phenoxy]propyI]thiazole4-carboxylic acid
Example 290: 2-[[6-(l,3-benzothiazol-2-yIamino)-5-methyl-pyridazin-3-yl]-methyl10 amino]-5-[3-[4-[3-[[(3B)-3,4-dihydroxybutyl]amino]prop-l-ynyl]-2-fluorophenoxy]propyl]thiazo!e-4-carboxylic acid
545
546
PHARMACOLOGICAL STUDY
EXAMPLE A: Fluorescence Polarisation Assay Data
Fluorescence polarization measures the rotation of a fluorescing species in solution, the larger the molécule the more polarized the fluorescence émission.
The fluorescent PUMA (UniProtKB® primary accession number Q9BXH1 - SEQ ID:01) based probe Fluorescein-betaAla-Ahx-AREIGAQLRRMADDLNAQY-OH from Biopeptides binds to GST(1-218)-(FACTOR XA)-/7sBCLXL(2-209) having an aminoacid sequence (SEQ ID:02):
[MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKKFELGLEFPNLPY YIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRYGVSRIAYSKDF ETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVLYMDPMCL DAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPKSDLIEGRGIPE FEFSQSNRELVVDFLSYKLSQKGYSWSQFSDVEENRTEAPEGTESEMETPSAINGNPS WHLADSPAVNGATGHSSSLDAREVIPMAAVKQALREAGDEFELRYRRAFSDLTSQL HITPGTAYQSFEQVVNELFRDGVNWGRIVAFFSFGGALCVESVDKEMQVLVSRIAAW MATYLNDHLEPWIQENGGWDTFVELYGNNAAAESRKGQER] (GST UniProtKB® primary accession number P08515 and BCLXL UniProtKB® primary accession number Q07817-1), resulting in an increase in anisotropy. If a compound is added which competitively binds to the same site as the probe, thereby releasing it, anisotropy decreases due to the increased amount of free probe.
An 11-point serial dilution of each compound was prepared in DMSO, the final buffer conditions were 10 mM 4-(2-hydroxyethyl)-l-piperazineethanesulfonic acid [HEPES], 150 mM NaCl, 0.05% Tween 20, pH 7.4 and 5% DMSO. The final protein concentration in the assay was 20 nM with the fluorescent probe présent at 10 nM. The experiments were incubated for 2 hours at 23°C before fluorescence polarization was measured on a Biotek SynergyNeo plate reader (Excitation 485nm, émission 525nm, parallel and perpendicular reads). The dose response curves were plotted with XL-Fit software using a 4-Parameter Logistic Model (Sigmoidal DoseResponse Model) and the inhibitory concentrations that gave a 50% increase in fluorescence intensity was determined (IC50). The Ki values were determined from the IC50
547 values according to Cer et al, Nucleic Acids Res, 2009, Jul l;37(WebServer issue): W441W445.
The results are summarised in Table 1. They show that the compounds of the invention inhibit the interaction between the Bcl-xL protein and the fluorescent peptide described hereinbefore.
Table 1
Example Ki (M) 22 2.9E-09 44 6.9E-08
1 3.1E-08 23 4.0E-08 45 1.4E-08
2 3.3E-07 24 1.4E-07 46 1.9E-07
3 1.9E-09 25 9.9E-07 47 2.3E-08
4 9.1E-07 26 6.5E-06 48 1.9E-06
5 1.8E-07 27 3.1E-07 49 8.1E-09
6 4.9E-07 28 1.3E-07 50 1.2E-09
7 7.2E-08 29 3.3E-08 51 6.4E-08
8 8.7E-08 30 8.1E-08 52 9.0E-08
9 4.0E-09 31 2.6E-09 53 1.5E-08
10 4.1E-07 32 2.6E-07 54 1.4E-09
11 6.0E-07 33 6.4E-08 55 8.2E-08
12 2.7E-08 34 8.9E-09 56 > 1E-05
13 3.8E-09 35 5.9E-08 57 3.5E-07
14 3.2E-09 36 2.8E-08 58 4.9E-08
15 3.7E-08 37 1.5E-08 59 < 1E-09
16 5.2E-08 38 2.5E-04 60 1.5E-09
17 2.2E-08 39 5.4E-08 61 < 1E-09
18 7.8E-08 40 2.0E-08 62 < 1E-09
19 3.0E-08 41 3.7E-08 63 < 1E-09
20 3.9E-08 42 4.7E-09 64 3.1E-08
21 7.5E-09 43 1.9E-08 65 < 1E-09
548
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
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 5.3E-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 1.3E-07
98 2.3E-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 1.3E-07
108 < 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 1.8E-07
146 < 1E-09
149 1.3E-07
150 < 1E-09
151 < 1E-09
156 8.9E-07
157 1.2E-06
158 4.7E-08
159 1.8E-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
170 < 1E-09
171 < 1E-09
172 < 1E-09
549
EXAMPLE B: AlphaLISA Assay Data
AlphaLISA assay measures an increase in alpha signal upon binding biotinylated GST(1-218)(FACTOR XA)-/?sBCLXL(2-209’) (SEQ ID:02 - GST UniProtKB® primary accession number P08515 and BCLXL UniProtKB® primary accession number Q07817-1) coupled to a Streptavidin labelled Alpha donor bead to N-terminally labelled peptide derived from BIM (Fluorescein-betaAla-Ahx- SEQ ID:05, wherein SEQ ID:05 is DMRPEIWIAQELRRIGDEANAYYARR) coupled to an Anti-FTIC AlphaLISA acceptor bead.
GST(1-218)-(FACTOR_XA)-/2sBÇLXL(2-209) was biotinylated in 50mM phosphate buffer using a 20 fold excess of EZ Link NHS-LC-Biotin (thermoFisher catalogue number 21336) overnight at 4°C.
The addition of a compound which binds competitively to the same site as the peptide will resuit in an decrease in the Alpha signal of the available donor bead-protein-peptide-acceptor bead complex due to displacement of the peptide.
An 11-point serial dilution of each compound was prepared in DMSO, the final buffer conditions were 10 mM 4-(2-hydroxyethyl)-l-piperazineethanesulfonic acid [HEPES], 150 mM NaCl, 0.05% Tween 20, pH 7.4 and 4% DMSO. The final protein concentration in the assay was 20pM with the peptide présent at 1 nM). Alpha beads were used at a final concentration of 5ug/mL. The experiments were incubated for overnight at 23 °C after acceptor bead addition. Finally the donor bead is added and incubated for 2 hours at 23 °C before Alpha signal measured on a Biotek Synergy2 plate reader (Excitation 680nm, émission 615nm). The dose response curves were plotted with XL-Fit software using a 4-Parameter Logistic Model (Sigmoidal DoseResponse Model) and the inhibitory concentrations that gave a 50% increase in fluorescence intensity was determined (IC50)·
The results are summarised in Table 2.
Table 2
Example IC50 (M)
3 9.6E-11
50 7.4E-12
54 2.5E-11
59 1.4E-10
60 9.6E-10
62 l.lE-10
63 l.OE-10
65 1.0E-09
73 7.7E-12
74 7.4E-11
76 7.7E-11
85 1.2E-08
89 1.5E-07
90 2.3E-11
550
91 5.3E-09
92 4.6E-08
93 6.4E-09
94 5.1E-08
95 2.2E-08
96 2.7E-07
97 2.1E-07
98 1.8E-09
100 3.2E-11
101 2.2E-12
108 6.6E-12
109 1.1E-11
110 4.8E-10
111 2.7E-08
112 2.8E-11
113 2.8E-11
114 5.5E-12
115 8.9E-11
116 1.7E-11
117 9.9E-12
118 1.8E-11
119 6.0E-12
120 9.9E-11
121 5.8E-11
122 9.8E-12
123 4.0E-12
124 7.6E-12
125 8.6E-09
126 2.3E-11
127 6.6E-12
128 1.3E-11
130 5.4E-12
131 2.7E-09
132 7.7E-09
133 8.9E-09
134 5.3E-12
135 3.9E-09
136 7.7E-09
137 2.7E-10
138 1.1E-08
139 2.7E-07
140 2.4E-11
141 1.4E-11
142 1.0E-08
143 2.2E-08
144 1.4E-11
145 1.9E-07
146 2.2E-11
147 7.1E-12
148 5.7E-12
150 1.3E-11
151 5.0E-12
152 2.3E-09
153 8.0E-08
161 3.2E-11
162 1.9E-09
163 1.8E-10
164 7.1E-12
165 1.9E-11
166 9.2E-12
167 1.3E-11
168 1.2E-11
169 9.9E-12
170 1.4E-11
171 1.3E-11
172 7.8E-12
173 7.9E-12
174 6.0E-10
551 φ^ΧΑΜΡΕΕ C: Quench Assay Data
Fluorescence quenching assay measures the change in fluorescence intensity of C-terminally Cy5labelled BCL-xL protein, His-His-(EK)-feBÇLXL(2-197)[N197C] (UniProtKB® primary accession number Q07817-1) having an amino acid sequence (SEQ ID:03):
[MHHHHHHHHGATGSTAGSGTAGSTGASGASTGGTGATHHHHHHHHDDDDKSPMGSQSN RELVVDFLSYKLSQKGYSWSQFSDVEENRTEAPEGTESEMETPSAINGNPSWHLADSPAVN GATGHSSSLDAREVIPMAAVKQALREAGDEFELRYRRAFSDLTSQLHITPGTAYQSFEQVVN ELFRDGVNWGRIVAFFSFGGALCVESVDKEMQVLVSRIAAWMATYLNDHLEPWIQENGG WDTFVELYG] which is linked in C-terminal région to the amino acid X which corresponds to a cysteine labelled on the sulfur with Sulfo-Cyanine5 from Lumiprobe GmbH catalogue number 13380, upon binding of a C-terminally labelled peptide derived from PUMA (UniProtKB® primary accession number Q9BXH1) having an aminoacid sequence (SEQ ID:04): [QWAREIGAQLRRMADDLNAQY] which is linked in C-terminal région to the amino acid X’ where X’ is cysteine labelled on the sulfur with TQ5WS from AAT Bioquest catalogue number 2079.
The addition of a compound which binds competitively to the same site as the peptide will resuit in an increase in the fluorescence intensity of the protein due to displacement of the fluorescence quencher.
An 11-point serial dilution of each compound was prepared in DMSO, the final buffer conditions were 10 mM 4-(2-hydroxyethyl)-l-piperazineethanesulfonic acid [HEPES], 150 mM NaCl, 0.05% Tween 20, pH 7.4 and 5% DMSO. The final protein concentration in the assay was 1 nM with the peptide présent at 400 nM. The experiments were incubated for 2 hours at 23°C before fluorescence intensity was measured on a Biotek SynergyNeo plate reader (Excitation 620nm, émission 680nm). The dose response curves were plotted with XL-Fit software using a 4-Parameter Logistic Model (Sigmoidal DoseResponse Model) and the inhibitory concentrations that gave a 50% increase in fluorescence intensity was determined (IC50)· The Ki values were determined from the IC50 values according to Cer et al, Nucleic Acids Res, 2009, Jul l;37(WebServer issue): W441-W445.
The results are summarised in Table 3.
Table 3
Example Ki (M)
50 4.4E-12
62 6.0E-12
552
73 2.8E-12
74 6.1E-12
101 5.2E-13
108 5.9E-13
109 1.1E-12
114 1.1E-11
116 8.6E-12
117 6.6E-12
118 3.3E-11
119 5.5E-13
122 3.0E-12
123 8.4E-13
124 1.7E-12
127 6.8E-13
128 2.8E-11
130 3.7E-13
134 1.0E-12
141 5.5E-12
144 3.9E-12
146 1.2E-12
147 9.7E-12
148 1.2E-12
150 1.7E-12
151 8.5E-13
163 2.0E-11
164 1.9E-12
165 5.9E-12
166 2.6E-12
167 2.9E-12
168 2.6E-12
169 4.4E-12
170 5.0E-12
171 3.5E-12
172 2.6E-12
173 3.3E-12
174 4.3E-11
175 1.1E-12
176 1.1E-12
177 3.0E-12
178 7.4E-12
179 6.8E-12
180 5.2E-12
181 4.3E-12
182 7.4E-12
183 7.8E-12
184 3.6E-12
185 1.2E-12
186 1.0E-12
187 2.5E-12
188 1.8E-12
189 2.4E-12
190 3.2E-12
191 2.8E-12
192 2.9E-12
193 1.5E-12
194 3.2E-12
195 1.8E-12
196 9.0E-13
197 1.3E-12
198 7.3E-12
199 1.0E-12
200 7.7E-12
201 1.9E-12
202 8.9E-13
203 2.2E-12
204 6.0E-12
205 2.3E-12
206 9.0E-12
207 3.7E-12
208 8.8E-12
209 2.3E-11
210 8.5E-13
211 6.5E-12
212 1.0E-12
213 1.5E-12
214 4.0E-12
215 3.9E-12
216 4.0E-12
217 4.1E-12
218 1.2E-12
219 8.2E-13
553
220 3.5E-12
221 1.7E-12
222 1.4E-12
223 1.8E-12
224 2.0E-12
225 3.5E-12
226 5.3E-12
227 3.5E-12
228 2.2E-10
229 1.4E-12
230 1.4E-12
231 1.3E-12
232 1.7E-12
233 4.3E-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 3.0E-12
271 3.3E-12
272 2.3E-12
The results of Tables 1-3 show that the compounds of the invention are potent inhibitors of the Bcl-xL protein.
EXAMPLE D: Effect of Bcl-xL inhibitors in MOLT-4 or H146 cell viability using MTT assay
MTT colorimétrie assay is based on the mitochondrial réduction of tétrazolium sait by living cells. The viable cell number is proportional to the production of formazan salts, which can be read spectrophotometrically at 540 nm.
MOLT-4 and H146 cells were purchased from ATCC and cultivated in RPMI1640 supplemented with 10 10% heat inactivated fêtai bovine sérum, penicillin (100 lU/ml), streptomycin (100 pg/ml) and Lglutamine (2 mM). Cells were cultured at 37°C in a humidified atmosphère containing 5% CO2.. Cells were seeded in 96 microwell plates (150pL per well) and exposed to the compounds for 48h (3,16 fold serially diluted; 9 concentrations each, triplicates). At the end of incubation time, 15 pL of MTT solution (5mg/ml) were added per well and the cells were incubated for another 4h. Then, lOOpL of 15 10% Sodium Dodecyl Sulfate (SDS)/HC1 lOmM were added per well and the plate was incubated overnight, before measurement of optical density at 540 nm. IC50S were calculated using standard fourparametric curve fitting. IC50 is defined as the compound concentration at which the MTT signal is reduced to 50% of that measured for the control. Results represent the mean of at least 2 independent experiments and are presented in Table 4 below.
554 ^Èable 4
Example H146 IC50 (M) MOLT-4 IC50 (M)
1 >1.5E-05 #N/A
2 1.25E-05 #N/A
3 4.2E-08 9E-08
5 8.21E-06 #N/A
6 >=1.49E-05 #N/A
7 >1.5E-05 #N/A
8 1.05E-05 #N/A
9 2.07E-07 2.32E-06
10 8.31E-06 #N/A
12 9.13E-07 1.62E-06
13 2.92E-07 2.23E-06
14 2.11E-06 #N/A
15 >1.5E-05 #N/A
16 8.04E-06 #N/A
17 1.34E-05 7.88E-06
18 >1.5E-05 #N/A
19 >=1.28E-05 #N/A
20 3.35E-06 7.21E-06
21 9.58E-06 #N/A
22 2.13E-06 #N/A
23 >=1.27E-05 1.3E-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
28 >1.5E-05 >1.5E-05
29 >1.5E-05 >1.5E-05
30 >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
555
52 1.3E-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
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 2,05 E-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 7.1 E-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
556
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 l,13E-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 #N/A 1.94E-07
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 4.3E-07
166 1.42E-07 1.72E-07
167 4.32E-07 4E-07
168 #N/A 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 1.62E-06 8.79E-07
197 4.39E-08 #N/A
198 1.42E-08 1.59E-09
557
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 7.3E-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 #n/a 7.52E-07
227 #n/a 8.02E-07
228 >3E-05 #N/A
229 4.37E-07 1.64E-06
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 #N/A 7.5E-07
238 #N/A 1.49E-07
239 #N/A 3.14E-07
240 #N/A 1.32E-06
241 #N/A 1.19E-06
242 #N/A 5.25E-07
243 #N/A 1.12E-07
247 #N/A 2.81E-09
250 #N/A 2.46E-10
253 #N/A 3.17E-08
267 #N/A 4.38E-08
268 #N/A 2.55E-10
269 #N/A 1.82E-10
270 #N/A 1.99E-10
271 #N/A 3.08E-08
272 #N/A 2.84E-10
558
These data show that the majority of these compounds are active in cells and can induce a dose dépendent decrease in the viability of H146 and Molt-4 cell lines.
EXAMPLE E: Pharmacodynamies and tumor régression study
The in vivo therapeutic and pharmacodynamie effects of Bcl-xL-targeting small molécules were determined in MOLT-4 T-cell Acute Lymphoblastic Leukemia (T-ALL) model upon intravenous (IV) administration. The tested compounds were formulated in Hydroxypropyl-P-Cyclodextrin 20%/Hydrogen Chloride 25 mM (HPBCD/HC1).
Materials and methods
MOLT-4 cells (ATCC No. CRL-1582) were cultured in RPMI supplemented with 10% FBS. Cells were re-suspended in 50% matrigel (BD Biosciences) and 0.1 mL containing 5xl06 cells was subcutaneously inoculated into the right flank of female NOD SCID mice (Charles River). For efficacy studies, when tumors reached the appropriate volume, mice were randomized (8 animais per group) using Easy stat software. Control vehicle (HPBCD/HC1), Example 73 (3.75 or 7.5 mg/kg) or Example 100 (3.75 mg/kg) were injected IV (twice weekly for 3 weeks - Q3D6). Mice body weight was monitored three times a week and tumor size was measured using electronic calipers. Tumor volume was estimated by measuring the minimum and maximum tumor diameters using the formula: (minimum diameter)2(maximum diameter)/2. At the end of the treatment cycle (day 17), tumor growth inhibition was calculated using the formula:
( Médian (TV at Dx in treated group) Ί
I--“— X lUv ( Médian (TV at Dx in Control group) )
Response was evaluated as follows: CR (Complété Response) if tumor size was <25mm3 for at least three consecutive measurements, PR (Partial Response) if tumor size was comprised between 25mm3 and half of the starting size for at least three consecutive measurements. Mice were sacrificed at the first measurement for which tumor volume exceeded 2000 mm3 or at the first signs of animal health détérioration.
For pharmacodynamies studies, when tumors reached the appropriate volume, mice were randomized (3 animais per group) using Easy stat software. Control vehicle (HPBCD/HC1) or Bcl-xL-targeting
559
Rnall molécules were injected IV (once per day - QD) at 7.5 mg/kg. Tumor samples were collected 6 h after dosing and lysed (10 mM HEPES pH 7.4, 142.5 mM KC1, 5 mM MgCh, 1 mM EDTA, 1% NP40, protease and phosphatase inhibitors cocktails - Calbiochem). Cleared lysâtes were prepared for immunodetection of cleaved PARP and Caspase 3 by using the MSD apoptosis panel whole cell lysate kit (MSD) in 96-well plates according to manufacturer’s instructions, and were analyzed on the QuickPlex SQ 120. Whole blood samples were analyzed on the Hematology Analyzer Coulter Ac»T diff (Beckman Coulter).
Ail experiments were conducted in accordance with the French régulations in force after approval by Servier Research Institute (IdRS) Ethical Committee. NOD SCID mice were maintained according to institutional guidelines.
Results
Efficacy of Example 73 and Example 100 on MOLT-4 xenografts is illustrated in Table 5. Treatment was started 12 days post tumor cells inoculation (average size: 214 mm3). Vehicle (HPBCD/HC1), Example 73 (3.75 and 7.5 mg/kg) or Example 100 (3.75 mg/kg) were dosed IV each 3 days for a total of 6 administrations.
On day 17 after treatment start, the Tumor Growth Inhibition (%TGI) of Example 73 was 56.9% at 3.75 mg/kg and 83.3% at 7.5 mg/kg (p<0.05), as depicted 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 relevant body weight loss due to the treatment was observed.
Table 5: MOLT-4 tumor growth inhibition upon treatment with Example 73 (3.75 and 7.5 mg/kg, administered IV, Q3D6) or Example 100 (3.75 mg/kg, administered IV, Q3D6).
Compound tested Dose (mg/kg) %TGI (day 17)
560
Example 73 3.75 56.9*
Example 73 7.5 83.3*
Example 100 3.75 64.6*
*p value <0.05 compared to control group.
The effect of Bcl-xL-targeting small molécules on apoptosis induction in MOLT-4 tumor cells and number of circulating platelets is illustrated in Table 6. Treatment was started 18 days post tumor cells inoculation (average size: 349 mm3). Vehicle (HPBCD/HC1), Example 74, Example 76 and Example (7.5 mg/kg) were dosed once IV and samples were collected 6 h later. Ail the compounds showed an induction of apoptosis markers, namely cleaved PARP (between 14.2 — 26.6-fold over control) and cleaved Caspase 3 (between 4.7 - 5.1-fold over control) cleavage. In addition, given the well-described rôle of Bcl-xL in regulating platelets life-span, ail tested compounds caused a significant réduction in platelet numbers (to 2 - 4% of control values).
Table 6: Cleaved PARP and Cleaved Caspase 3 in tumor cells and platelet loss in MOLT-4grafted female NOD SCID mice 6 h after treatment with indicated compounds at 7.5 mg/kg, administered IV.
Compound tested Dose (mg/kg) and route Timepoint (h) Cleaved PARP (fold increase over control) Cleaved Caspase 3 (fold increase over control) Platelet count (χ103/μ1) % remaining platelets (vs vehicle)
Example 74 7.5, IV 6 17.1 5.1 18 2
Example 76 7.5, IV 6 14.2 4.9 40 3
Example 73 7.5, IV 6 21.4 4.7 51 4
In conclusion, we show here that the Bcl-xL-targeting small molécules described in Examples 73, 74, 15 76 and 100 are active in vivo after intravenous administration. We observed tumor régression, apoptosis induction in tumor cells and a strong réduction in circulating platelets, in agreement with the previously described rôle of Bcl-xL in apoptosis control and platelets life-span régulation (Youle and
561
Btrasser, Nat. Rev. Mol. Cell Biol. 2008 Jan;9(l):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 relevant body weight loss was observed upon treatment with efficacious doses and platelets loss was recovered after treatment discontinuation (data not shown). Altogether, these data indicate that there is a possible therapeutic margin for the use of these Bcl-xL-targeting small molécules in cancer treatment.
EXAMPLE F: In vivo pharmacodynamie profile of the compounds of formula (I)
The pharmacokinetic profile of the compounds of formula (I) is evaluated in rodent (mouse, rat) after PO and/or IV route. The formulation is selected based on the physico-chemical properties of the tested drug as well as the route of administration. A single dose of the drug (<5mg/kg) prepared in the adapted formulation is administered by IV (bolus or 10min infusion) or PO (gavage) route to animais (3 animals/route). Blood samples from each animal (up to 6 samples/animal) are collected over 24 h after dosing and plasma concentrations of the tested compound are determined after extraction followed by a liquid chromatography coupled with tandem mass spectrometry détection (LC/MS-MS).
In some cases, the following experimental protocol is used to détermine the pharmacokinetic profile of the compounds of the invention in the Wistar rat:
The drug is prepared in a formulation composed by a mixture of polyethylene glycol 300/anhydrous Ethanol/NaCl 0.9% (40/10/50 v/v/v). The formulation is administered by IV route to male Wistar rat (3 animais) at a dose of 0.75mg/kg (10min inf, 5mL/kg). Blood samples are taken at the following time points from each animal: end of infusion (10min), 0.5h, lh, 3h, 6h and 24h after dosing. The plasma concentrations of the tested compound are determined after extraction followed by a liquid chromatography coupled with tandem mass spectrometry détection (LC/MS-MS).
The lower limit of quantification is 2.5ng/mL.
The results allow to rank the compounds of the invention based on their plasma exposure, élimination rate constant, clearance and volume of distribution in order to evaluate the therapeutic range of the compounds in animal model.

Claims (34)

  1. wherein:
    ♦ Ri and R2 independently of one another represent a group selected from: hydrogen; linear or branched Ci-Côalkyl optionally substituted by a hydroxyl or a Ci-Côalkoxy group; C3-C6cycloalkyl; trifluoromethyl; linear or branched Ci-Côalkylene-heterocycloalkyl wherein the heterocycloalkyl group is optionally substituted by a a linear or branched Ci-Cealkyl group;
    or Ri and R2 form with the carbon atoms carrying them a C3-C6cycloalkylene group, ♦ R3 represents a group selected from: hydrogen; C3-C6cycloalkyl; linear or branched Ci-Côalkyl; -Xi-NRaRb; -Xi-N+RaRbRc; -Xi-O-Rc; -Xi-COORc; -Xi-PO(OH)2; -Xi-SO2(OH); -X1-N3 and :
    -Χή-=CH ?
    ♦ Ra and Rb independently of one another represent a group selected from: hydrogen; heterocycloalkyl; -SO2-phenyl wherein the phenyl may be substituted by a linear or branched Ci-Côalkyl; linear or branched Ci-Cealkyl optionally substituted by one or two hydroxyl groups; Ci-C6alkylene-SO2OH; Ci-C6alkylene-SO2O·; Ci-Cealkylene-COOH; Ci-CealkylenePO(OH)2; Ci-Côalkylene-NRdRe; Ci-C6alkylene-N+RdReRf; Ci-Cealkylene-phenyl wherein the phenyl may be substituted by a Ci-Côalkoxy group;
    the group:
    or Ra and Rb form with the nitrogen atom carrying them a cycle Bi;
    563
    or Ra, Rb and Rc form with the nitrogen atom carrying them a bridged Cs-Csheterocycloalkyl, ♦ Rc, Rd, Re, Rf, independently of one another represents a hydrogen or a linear or branched Ci-
    Côalkyl group, or Ra and Re form with the nitrogen atom carrying them a a cycle B2, or Rd, Re and Rf form with the nitrogen atom carrying them a bridged Ca-Csheterocycloalkyl, ♦ Heti represents a group selected from:
    ♦ Het2 represents a group selected from:
    564
    ♦ Ai is -NH-, -N(Ci-C3alkyl), O, S or Se, ♦ A2 is N, CH or C(R5), ♦ G is selected from the group consisting of:
    -C(O)ORG3, -C(0)NRgiRg2, -C(O)Rg2, -NRgiC(O)RG2, -NRgiC(O)NRgiRg2,
    5 -OC(O)NRgiRg2, -NRgiC(O)ORG3, -C(=NORgi)NRgiRg2,
    -NRgiC(=NCN)NRgiRg2, -NRgiS(O)2NRgiRg2, -S(O)2Rg3, -S(O)2NRGiRG2,
    -NRgiS(O)2RG2, -NRgiC(=NRG2)NRgiRg2, -C(=S)NRGiRg2, -C(=NRGi)NRgiRG2, halogen, -NO2, and -CN, in which:
    - Rgi and Rg2 at each occurrence are each independently selected from the group consisting of 10 hydrogen, Ci-Côalkyl optionally substituted by 1 to 3 halogen atoms, C2-Côalkenyl, C2-Côalkynyl, Cs-Côcycloalkyl, phenyl and -(CH2)i-4-phenyl;
    - Rg3 is selected from the group consisting of Ci-Côalkyl optionally substituted by 1 to 3 halogen atoms, C2-C6alkenyl, C2-Côalkynyl, Cs-Côcycloalkyl, phenyl and -(CH2)i-4-phenyl; or
    Rgi and Rg2, together with the atom to which each is attached are combined to form a C315 Csheterocycloalkyl ; or in the alternative, G is selected from the group consisting of:
    565
    wherein Rg4 is selected from Ci-Cealkyl optionally substituted by 1 to 3 halogen atoms, C2Côalkenyl, C2-Côalkynyl and Cs-Côcycloalkyl, ♦ R4 represents a hydrogen, fluorine, chlorine or bromine atom, a methyl, a hydroxyl or a 5 methoxy group, ♦ R5 represents a group selected from: Ci-Côalkyl optionally substituted by 1 to 3 halogen atoms; C2-C6alkenyl; C2-Côalkynyl; halogen or -CN, ♦ Rô represents a group selected from:
    hydrogen;
    10 -C2-C6alkenyl;
    -X2-O-R7;
    o—r7
    -X2-NSO2-R7;
    -C=C(R9)-Yi-O-R7;
    15 Cs-Côcycloalkyl;
    Cs-Côheterocycloalkyl optionally substituted by a hydroxyl group;
    C3-C6cycloalkylene-Y2-R7 ;
    566
    Cs-Côheterocycloalkylene-Yz-R? group, an heteroarylene-R? group optionally substituted by a linear or branched Ci-Cealkyl group, ♦ R? represents a group selected from: linear or branched Ci-Cealkyl group;
    (Cs-Cejcycloalkylene-Rs; or:
    wherein Cy represents a C3-C8cycloalkyl, ♦ R8 represents a group selected from: hydrogen; linear or branched Ci-Cealkyl, -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;
    - 0-X’2-NR’aR’b, -X’2-NR’aR’b, -NR’C-X’2-N3 and :
    -NR'C—X’?^=C H ?
    ♦ R9 represents a group selected from linear or branched Ci-Côalkyl, trifluoromethyl, hydroxyl, halogen, Ci-Cealkoxy, ♦ Rio represents a group selected from hydrogen, fluorine, chlorine, bromine, -CF3 and methyl,
    567 ♦ Ru represents a group selected from hydrogen, Ci-C3alkylene-R8, -O-Ci-C3alkylene-R8, -CONRhRi and -CH=CH-Ci-C4alkylene-NRhRi, -CH=CH-CHO, C3-C8cycloalkylene-CH2-R8, C3C8heterocycloalkylene-CH2-R8, ♦ R12 and Ri3, independently of one another, represent a hydrogen atom or a methyl group, ♦ Ri4 and Ris, independently of one another, represent a hydrogen or a methyl group, or Rj4 and Ris form with the carbon atom carrying them a a cyclohexyl, ♦ Rh and Ri, independently of one another, represent a hydrogen or a linear or branched CiCealkyl group, ♦ Xi and X2 independently of one another, represent a linear or branched Ci-Cealkylene group optionally substituted by one or two groups selected from trifluoromethyl, hydroxyl, halogen, Ci-Côalkoxy, ♦ X’2 represents a linear or branched Ci-Côalkylene, ♦ R’a and R’b independently of one another, represent a group selected from: hydrogen; heterocycloalkyl; -SO2-phenyl wherein the phenyl may be substituted by a linear or branched Ci-Cealkyl; linear or branched Ci-Côalkyl optionally substituted by one or two hydroxyl or CiCôalkoxy groups; Ci-C6alkylene-SO2OH; Ci-Csalkylene-SChO; Ci-Côalkylene-COOH; Ci-C6alkylene-PO(OH)2; Ci-C6alkylene-NR’dR’e; Ci-Cealkylene-N+R’dR’eR’f; Ci-Côalkylene-O-Ci-Côalkylene-OH; Ci-Câalkylene-phenyl wherein the phenyl may be substituted by a hydroxyl or a Ci-Côalkoxy group;
    the group:
    or R\ and R’b form with the nitrogen atom carrying them a cycle B3, or R’a, R’b and R’c form with the nitrogen atom carrying them a bridged
    C3-C8heterocycloalkyl, ♦ R’c, R’d, R’e, R’f, independently of one another, represents a hydrogen or a linear or branched Ci-Côalkyl group, or R’d and R’e form with the nitrogen atom carrying them a cycle B4,
    568 or R’d, R’e and R’f form with the nitrogen atom carrying them a bridged Cs-Csheterocycloalkyl, ♦ Yi represents a linear or branched Ci-C4alkylene, ♦ Y2 represents a bond, -O-, -O-CH2-, -O-CO-, -O-SO2-, -CH2-, -CH2-O, -CH2-CO-, -CH2-SO2-,-C2H5-, -CO-, -CO-O-, -CO-CH2-, -CO-NH-CH2-, -SO2-, -SO2-CH2-, -NH-CO-, -NH-SO2-, ♦ m=0,1 or 2, ♦ p=l, 2, 3 or 4, ♦ Bi, B2, B3 and B4, independently of one another, represents a Cs-Csheterocycloalkyl group, which group can: (i) be a mono- or bi-cyclic group, wherein bicyclic group includes fused, briged or spiro ring System, (ii) can contain, in addition to the nitrogen atom, one or two hetero atoms selected independently from oxygen, sulphur and nitrogen, (iii) be substituted by one or two groups selected from: fluorine, bromine, chlorine, linear or branched Ci-Côalkyl, hydroxyl, -NH2, oxo or piperidinyl, its enantiomers and diastereoisomers, and addition salts thereof with a pharmaceutically acceptable acid or base.
  2. 2. Compound according to claim 1 wherein Heti represents:
  3. 3. Compound according to claim 1 or 2 wherein Ri represents a methyl group and R2 represents a hydrogen atom.
    4, Compound according to claim 1 or 2 wherein Ri and R2 represent each a methyl group or a hydrogen atom.
  4. 5. Compound according to claim 1 or 2 wherein Ri or R2 represents a linear or branched Ci-
    569
    Eôalkyl optionally substituted by a hydroxyl or a Ci-Côalkoxy group, preferably a group selected from: methyl, methoxymethyl, hydroxymethyl, ethyl and prop-2-yl.
    & Compound according to claim 1 or 2 wherein Ri or R2 represents a Cs-Cecycloalkyl, preferably a cyclopropyl group.
    L Compound according to claim 1 or 2 wherein Ri represents a trifluoromethyl group.
  5. 8. Compound according to claim 1 or 2 wherein R2 represents a linear or branched Ci-Côalkylene- heterocycloalkyl, preferably a (4-methyl-piperazin-l-yl)propyl group.
  6. 9. Compound according to claim 1 or 2 wherein Ri and R2 form with the carbon atoms carrying them a cyclohexene or a cyclopentene group.
  7. 10. Compound according to any of claims 1 to 9 wherein R3 represents a hydrogen atom or a methyl group.
  8. 11. Compound according to any of claims 1 to 9 wherein R3 represents -Xi-PO(OH)2, -XiSO2(OH), -Xi-NRaRb; -Xi-N+RaRbRc, wherein Ra or Rb, or both of them, represent a group selected from Ci-C6alkylene-SO2OH, Ci-C6alkylene-SO2O' and Ci-C6alkylene-PO(OH)2.
    12, Compound according to any of claims 1 to 9 wherein Rg represents -NR’aR’b; -N+R’aR’bR’c; NH-X’2-N+R’aR’bR’c, wherein R’a and R’b, or both of them, represent a group selected from CiC6alkylene-SO2OH and Ci-Côalkylene-PO(OH)2.
  9. 13. Compound according to any of claims 1 to 9 wherein R3 represents a group selected from: ethyl, propyl; 2-methoxy-ethyl, 2-(morpholin-4-yl)ethyl, 3-(morpholin-4-yl)propyl, 3-diethylaminopropyl, 3-methoxy-propyl, 3-hydroxy-propyl, 3,4-dihydroxy-butyl, 4-methoxy-3-hydroxy-butyl, 4hydroxy-3-methoxy-butyl, 2,3-dihydroxy-propyl, 4,5-dihydroxy-pentyl, 4-hydroxy-butyl, 3-hydroxy2-morpholino-propyl, 5-hydroxy-4-methoxy-pentyl, 5-morpholino-4-hydroxy-pentyl, 3-hydroxy-2methoxy-propyl, 5-[2-(dimethylamino)ethyl-methyl-amino]-4-hydroxy-pentyl, 5-hydroxy-pentyl, 5methoxy-4-hydroxy-pentyl, 5-(dimethylamino)-4-hydroxy-pentyl, 4-hydroxy-5- (trimethylammonio)pentyl, 5-[3-sulfonate-propyl-dimethyl-ammonio]-4-hydroxy-pentyl, 4-hydroxy5-(methylamino)pentyl, 3-carboxy-propyl, 5-[methyl(4-piperidyl)amino]pentyl, 5-(220487
    570
    Riorpholinoethylamino)pentyl, 5-[2-(4-methyl-piperazin-l-yl)ethylamino)pentyl, 4-[2-(4-methylpiperazin-1 -yl)ethylamino)butyl, 4-hydroxy-5- [methyl- [2-(methylamino)ethyl] amino]pentyl, 5 -[2(diethylamino)ethylamino]-4-hydroxy-pentyl, 5-(4-Amino-l-piperidyl)-4-hydroxy-pentyl, 4hydroxy-5-piperazin-1 -yl-penty 1,5-[2-(l -piperidy l)ethylamino]pentyl, 4-(4-Amino-1 -piperidy l)butyl, 4- [2-(diethy lamino)ethylamino]buty 1, 5 -(4-Amino- 1-piperidy l)penty 1, 4-[methyl-[2(methy lamino)ethy l]amino]buty 1, 4-(2-morpholinoethylamino)butyl, 4- [2-(4-methyl-piperazin-1 yl)ethylamino)butyl, 4-[(l-methyl-4-piperidyl)amino]butyl, 5-[2-(diethylamino)ethylamino]pentyl, 4piperazin-1 -ylbuty 1,4-(methylamino)butyl, 5 -piperazin- 1-ylpentyl, 5 - [methyl- [2-(methy lamino)ethy 1, 5-aminopentyl, 4-aminobutyl, 5-(methylamino)pentyl, 5-[3-(4-methylpiperazin-lyl)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-sulfonate-propyldimethy l-ammonio]pentyl, 4-(trimethy lammonio)buty 1, 4-hydroxy-5-quinuclidin-1 -ium-1 -yl-penty 1, 4-hydroxy-5-(l-aza-4-azoniabicyclo[2.2.2]octan-4-yl)pentyl, 4-hydroxy-5-(4-methylmorpholin-4ium-4-y l)pentyl, 5 -(trimethylammonio)pentyl, 4-hydroxy-5-(l -methylpiperidin-1 -ium-1 -y l)penty 1, 5(l,4-dimethylpiperazin-l-ium-l-yl)-4-hydroxy-pentyl, 6-(trimethylammonio)hexyl, 5-[3hydroxypropyl(dimethyl)ammonio]pentyl, 5-[2-hydroxyethyl(dimethyl)ammonio]pentyl, 5[carboxymethy l(dimethyl)ammonio]penty 1, 5 - [carboxymethyl(dimethy l)ammonio]penty 1, 3 phosphonopropyl, 3-[3-sulfonate-propyl-dimethyl-ammonio]propyl, 3-sulfopropyl, 4-sulfobutyl, 4phosphonobutyl, 4-carboxybutyl, 3-aminopropyl, 3-azidopropyl, pent-4-yn-l-yl, 4-aminobutyl, 4azidobutyl, hex-5-yn-l-yl, 5-azidopentyl.
  10. 14. Compound according to any of daims 1 to 13 wherein Het2 represents:
    15, Compound according to any of daims 1 to 13 wherein Het? represents:
    COOH
    16, Compound according to claim 14 or 15 wherein Rô represents a hydrogen atom.
    571
    Compound according to claim 14 wherein Rô represents a -X2-O-R7 group wherein X2 is a propylene group.
  11. 18. Compound according to claim 17 wherein R7 represents the following group:
  12. 19. Compound according to claim 17 wherein R7 represents the following group:
  13. 20. Compound according to claim 18 or 19 wherein Rs represents a group selected from: dimethylamino, methylamino, methylethylamino, diethylamino, methyl[2(methylamino)ethyl] amino, (2-hydroxyethyl)(methyl)amino, 4-morpholinyl, pyrrolidin-l-yl, 1piperidyl, [(tert-butoxy)carbonyl](methyl)amino, hydroxyl, bis(3-sulfopropyl)amino, 310 sulfopropylamino, methyl(3-sulfopropyl)amino, methyl(p-tolylsulfonyl)amino, (4methoxyphenyl)methyl-methyl-amino, 2-(dimethylamino)ethylamino and :
    21, Compound according to claim 18 or 19 wherein Rs represents a group selected from: 315 piperazin-l-yl, 4-methylpiperazin-l-yl, 3-hydroxypropyl(methyl)amino, [(35)-3,4-dihydroxybutyl]methyl-amino, 3-hydroxypropylamino, [(35)-3,4-dihydroxybutyl]amino, 4- hydroxybutyl(methyl)amino, [(3R)-3,4-dihydroxybutyl]-methyl-amino, 4-hydroxybutylamino, [(3R)3,4-dihy droxybuty l]amino.
  14. 22. Compound according to claim 17 wherein R7 represents:
    572
    wherein Ru is selected from dimethylcarbamoyl, 3-(dimethylamino)propyl, 3-(methylamino)propyl, 3-(methylamino)propyl.
  15. 23. Compound according to claim 14 wherein Re represents a C3-C6heterocycloalkylene-Y2-R7 5 group wherein the heterocycloalkylene group is selected from:
    Y2—R7
  16. 24. Compound according to claim 23 wherein R? is selected from: methyl, methylethyl, ZerZ-butyl, 2-methylpropyl and phenyl, preferably phenyl.
  17. 25. Compound according to claim 14 or 15 wherein Rô represents -C=C(Rg)-Yi-O-R7 wherein Yi is a methylene group.
    10
  18. 26. Compound according to claim 14 or 15 wherein Rô represents:
  19. 27. Compound according to claim 26 wherein R? represents a group selected from :
    573
    wherein Rs represents a group selected from: hydrogen, 2-(methylamino)ethoxy and:
    2& The compound according to claim 26 wherein R? represents a group selected from :
    wherein Rs 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-l-ylethoxy, 2-(410 methylpiperazin-l-yl)ethoxy, 2-(4-hydroxybutylamino)ethoxy, 2-piperazin-l-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.
  20. 29. A compound according to claim 1 selected in the following group:
    - 2-({6-[(l,3-Benzothiazol-2-yl)ammo]-5-methylpyridazin-3-yl}(4-hydroxybutyl)amino)-5-(315 {2-fluoro-4-[3-(methylammo)prop-l-yn-l-yl]phenoxy}propyl)-l,3-thiazole-4-carboxylic
    574 acid, 2-[[6-(l,3-Benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-methyl-amino]-5-[3-[4-[3(dimethylamino)prop-1 -y nyl] -2-fluoro-phenoxy]propyl] thiazole-4-carboxylic acid, 2-({6-[(l,3-Benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)-5-(3-{2fluoro-4-[3 -(methy lamino)prop-1 -yn-1 -yl]phenoxy }propy 1)-1,3-thiazole-4-carboxylic acid, 2-[[6-(l,3-Benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-(3,4-dihydroxybutyl)amino]-5[3-[2-fluoro-4-[3-(methylamino)prop-l-ynyl]phenoxy]piOpyl]thiazole-4-carboxylic acid, 2-[[6-(l,3-Benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-(3-hydroxy-4-methoxybutyl)amino]-5-[3-[2-fluoro-4-[3-(methylamino)prop-l-ynyl]phenoxy]propyl]thiazole-4carboxylic acid,
    - 2-[[6-(l,3-Benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-(2,3-dihydroxypiOpyl)amino]5 -[3 - [2-fluoro-4- [3-(methylamino)prop-l -ynyl]phenoxy]propy 1] thiazole-4-carboxylic acid, 2-[[6-(l,3-Benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-(4,5-dihydiOxypentyl)amino]5-[3-[2-fluoiO-4-[3-(methylamino)prop-l-ynyl]phenoxy]propyl]thiazole-4-carboxylic acid, 2-[[6-(l,3-Benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-(3,4-dihydroxybutyl)amino]-5[3-[4-[3-(dimethylamino)prop-l-ynyl]-2-fluoro-phenoxy]propyl]thiazole-4-carboxylic acid, 2-[[6-(l,3-Benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-(3-hydroxypropyl)amino]-5-[3[2-fluoro-4-[3-(methylamino)prop-l-ynyl]phenoxy]propyl]thiazole-4-carboxylic acid, 2-[[6-(l,3-Benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-[4-hydroxy-5(trimethylammonio)pentyl] amino] -5-[3 - [2-fluoro-4- [3-(methy lamino)prop-1 ynyl]phenoxy]propyl]thiazole-4-carboxylate, ’ 3-[[5-[[6-(l,3-Benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-[4-carboxy-5-[3-[2-fluoro4- [3-(methylamino)prop-1 -yny l]phenoxy]propyl] thiazol-2-y 1] amino] -2-hy droxy-penty 1] dimethyl-ammonio]propane-l-sulfonate,
    - 2-[[6-(l,3-Benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-(4-hydroxybutyl)amino]-5-[3[2-fluoiO-4-[3-[methyl(3-sulfopropyl)amino]prop-l-ynyl]phenoxy]propyl]thiazole-4carboxylic acid, 5-{l-[(Adamantan-l-yl)methyl]-5-methyl-lH-pyrazol-4-yl}-2-({6-[(l,3-benzothiazol-2yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)-l,3-thiazole-4-carboxylic acid, 5-{l-[(Adamantan-l-yl)methyl]-5-methyl-l/Z-pyrazol-4-yl}-2-({6-[(l,3-benzothiazol-2yl)amino]-5-cyclopropylpyridazin-3-yl}(methyl)amino)-l,3-thiazole-4-carboxylic acid,
    - 2-[[6-(l,3-Benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-[5-[2- (diethylamino)ethylamino]-4-hydroxy-pentyl]amino]-5-[3-[4-[3-(dimethylamino)prop-lynyl]-2-fluoro-phenoxy]propyl]thiazole-4-carboxylic acid,
    575
    2- [[6-(1,3-Benzothiazol-2-ylamino)-5 -methyl-py ridazin-3 -y 1] - [5 - [2-(4-methy Ipiperazin-1 y l)ethy lamino]pentyl] amino] -5 - [3- [4-[3 -(dimethylamino)prop-1 -ynyl] -2-fluorophenoxy]propyl]thiazole-4-carboxylic acid,
    2- [[6-(l ,3-Benzothiazol-2-ylamino)-5 -methyl-py ridazin-3 -y 1] -(4-hy droxy -5 -piperazin-1 -y 1pentyl)amino]-5-[3-[4-[3-(dimethylamino)prop-l-ynyl]-2-fluoro-phenoxy]propyl]thiazole-4carboxylic acid,
    2-({ 6 - [( 1,3 -Benzothiazol-2-yl)amino] -5 -methy lpyridazin-3 -yl} (methy l)amino)-5 - [(25)-3- {4[3-(dimethy lamino)prop-1 -y n- 1-yl] -2-fluorophenoxy } -2-methy Ipropy 1] -1,3 -thiazole-4carboxylic acid,
    2- [4-Aminobutyl- [6-(l ,3 -benzothiazol-2-ylamino)-5 -methy 1-py ridazin-3 -y 1] amino] -5 - [3 - [4[3-(dimethylamino)prop-l-ynyl]-2-fluoro-phenoxy]propyl]thiazole-4-carboxylic acid,
    - 2-[5-Aminopentyl-[6-(l,3-benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]amino]-5-[3-[4[3-(dimethylamino)prop-l-ynyl]-2-fluoro-phenoxy]propyl]thiazole-4-carboxylic acid, 2-[[6-(l,3-Benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-[3(dimethylamino)propyl]amino]-5-[3-[2-fluoro-4-[3-(methylamino)prop-lynyl]phenoxy]propyl]thiazole-4-carboxylic acid,
    2-[[6-(l,3-Benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-methyl-amino]-5-[3-[4-[3(dimethylamino)but-l-ynyl]-2-fluoro-phenoxy]propyl]thiazole-4-carboxylic acid, 2-[[6-(l,3-Benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-[4-hydroxy-5(trimethylammonio)pentyl]amino]-5-[3-[4-[3-(dimethylamino)prop-l-ynyl]-2-fluorophenoxy]propyl]thiazole-4-carboxylate,
    - 3-[2-[3-[4-[3-[2-[[6-(l,3-Benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-methyl-amino]4-carboxy-thiazol-5-yl]propoxy]-3-fluoro-phenyl]prop-2-ynylamino]ethyl-dimethylammonio]propane-1 -sulfonate,
    - 2-[[6-(l,3-Benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-[4-hydroxy-5-(4methy lmorpholin-4-ium-4-yl)penty 1] amino] -5 - [3- [4-[3-(dimethy lamino)prop-1 -ynyl] -2fluoro-phenoxy]propyl]thiazole-4-carboxylate,
    - 2-[3-[4-[3-[2-[[6-(l,3-Benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-methyl-amino]-4carboxy-thiazol-5-yl]propoxy]-3-fluoro-phenyl]prop-2-ynylamino]ethyl-trimethylammonium,
    2-[[6-(l,3-Benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-[6(trimethy lammonio)hexy 1] amino] -5 - [3 -[2-fluoro-4- [3 -(methy lamino)prop-1 ynyl]phenoxy]propyl]thiazole-4-carboxylate,
    - 2-[[6-(l,3-Benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-[5-[2-
    576 hy droxyethyl(dimethyl)ammonio]pentyl] amino] -5 - [3 - [4- [3-(dimethylamino)prop-1 -ynyl] -2fluoro-phenoxy]propyl]thiazole-4-carboxylate,
    2 - [[6-(l ,3-Benzothiazol-2-ylamino)-5 -methyl-pyridazin-3 -yl] - [5 -
    [carboxymethyl(dimethyl)ammonio]pentyl]amino]-5-[3-[4-[3-(dimethylamino)prop-l-ynyl]2-fluoro-phenoxy]propyl]thiazole-4-carboxylate,
    3-[[5-[[6-(l,3-Benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-[4-carboxy-5-[3-[2-fluoro-
    4 - [(E)-3-oxoprop-1 -enyl]phenoxy ]propyl] thiazol-2-y 1] amino] -2-hy droxy-pentyl] -dimethylammonio]propane-1 -sulfonate, 2-[[6-(l,3-Benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-methyl-amino]-5-[3-[4-[3(dimethy lamino)prop-1 -ynyl]phenoxy]propy 1] thiazole-4-carboxylie acid,
    - 2-[[6-(l,3-Benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-methyl-amino]-5-[3-[4-[l[(dimethylamino)methyl]-3-bicyclo[7.7.7]pentanyl]phenoxy]propyl]thiazole-4-carboxylic acid,
    - 2-[[6-(l,3-Benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-(3-phosphonopropyl)amino]-5[3-[2-fluoro-4-[3-(methylamino)prop-l-ynyl]phenoxy]propyl]thiazole-4-carboxylic acid,
    - 2-({ 6-[(l ,3 -Benzothiazol-2-y l)amino] -5 -me thylpy ridazin-3-yl} (3 -carboxypropy l)amino)-5 - (3-{4-[3-(dimethylamino)prop-l-yn-l-yl]-2-fluorophenoxy}propyl)-l,3-thiazole-4-carboxylic acid,
    - 2-({ 6-[(l ,3 -Benzothiazol-2-yl)amino] -5 -methy lpyridazin-3-yl} (3 -phosphonopropy l)amino)5 -(3- {4- [3 -(dimethy lamino)prop- 1-yn-1 -y 1] -2-fluorophenoxy }propyl)-1,3 -thiazole-4carboxylic acid,
    - 2-({ 6- [(1,3-Benzothiazol-2-yl)amino] -5 -methy lpyridazin-3-y 1 }(3-sulfopropy l)amino)-5 -(3 {4- [3 -(dimethylamino)prop-1 -yn-1 -yl] -2-fluorophenoxy }propyl)-1,3 -thiazole-4-carboxylic acid,
    - 2-({6-[(l,3-Benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(4-sulfobutyl)amino)-5-(3-{4[3-(dimethylamino)prop-1 -yn-1 -y 1] -2-fluorophenoxy }propyl)-1,3-thiazole-4-carboxylic acid,
    - 2-({6-[(l,3-Benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(4-phosphonobutyl)amino)-5(3-{4-[3-(dimethylamino)prop-l-yn-l-yl]-2-fluorophenoxy}propyl)-l,3-thiazole-4-carboxylic acid,
    - 2-({ 6- [(1,3-Benzothiazol-2-y l)amino] -5 -methy lpyridazin-3 -y 1} (4-carboxybuty l)amino)-5-(3 - {4- [3-(dimethylamino)prop-1 -yn-1 -yl] -2-fluorophenoxy }propyl)-1,3-thiazole-4-carboxy lie acid,
    - 2-[{6-[(l,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(pent-4-yn-l-yl)amino]-5-(3{2-fluoro-4- [3-(methy lamino)prop-l -yn-1 -yl]phenoxy }propy 1)-1,3 -thiazole-4-carboxylic
    577
    P acid,
    2-({ 6- [(1,3-Benzothiazol-2-yl)amino] -5 -methylpyridazin-3-yl} (methy l)amino)-5-{3-[2fluoro-4-(3 -{methy 1 [2-(methy lamino)ethy l]amino }prop- 1-yn-1 -y l)phenoxy ]propyl} -1,3thiazole-4-carboxylic acid, 2-({6-[(l,3-Benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)-5-[3-(2fluoro-4- { 3 - [(2-hy droxy ethy l)(methyl)amino]prop-1 -y η-1 -yl }phenoxy )propy 1] -1,3-thiazole4-carboxylic acid, 2-({6-[(l,3-Benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino)-5-{3-[2fluoro-4-(3 -hydroxyprop-1 -yn-1 -yl)phenoxy]propy 1} -1,3-thiazole-4-carboxylie acid, 2- [5-azidopenty 1- [6- [(1,3 -benzothiazol-2-y l)amino] -5 -methyl-py ridazin-3 -y 1] amino] -5-[3-[2fluoro-4-[3-(methylamino)prop-l-ynyl]phenoxy]propyl]-l,3-thiazole-4-carboxylic acid, its enantiomers and diastereoisomers, and addition salts thereof with a pharmaceutically acceptable acid or base.
  21. 30. A compound according to claim 1 selected in the following group:
    6-[{6-[(l,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino]-3-[l-({3-[2(dimethy lamino)ethoxy] -5,7-dimethyladamantan-1 -y 1 }methyl)-5 -methyl-1 H-py razol -4yl]pyridine-2-carboxylic acid,
    - 6-[[6-(l,3-benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-methyl-amino]-3-[l-[[3-[2-(3hydroxypropylamino)ethoxy]-5,7-dimethyl-l-adamantyl]methyl]-5-methyl-pyrazol-4yl]pyridine-2-carboxylic acid,
    - 6-[[6-(l,3-benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-methyl-amino]-3-[l-[[3-[2-(3methoxypropy lamino)ethoxy ] -5,7-dimethy 1 -1 -adamantyl]methyl] -5 -methy 1-py razol-4yl]pyridine-2-carboxylic acid, ' 6-[{6-[(l,3-benzothiazol-2-yl)amino]-5-methylpyridazin-3-yl}(methyl)amino]-3-(l-{[3-(2{[(35)-3,4-dihydroxybutyl]amino}ethoxy)-5,7-dimethyladamantan-l-yl]methyl}-5-methyll//-pyrazol-4-yl)pyridine-2-carboxylic acid,
    - 6-[[6-(l,3-benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-methyl-amino]-3-[l-[[3,5dimethyl-7-(2-morpholinoethoxy)-l-adamantyl]methyl]-5-methyl-pyrazol-4-yl]pyridine-2carboxylic acid,
    - 6-[[6-(l,3-benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-methyl-amino]-3-[l-[[3-[2-(2carboxyethylamino)ethoxy]-5,7-dimethyl-l-adamantyl]methyl]-5-methyl-pyrazol-4yl]pyridine-2-carboxylic acid,
    - 6-[[6-(l,3-benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-methyl-amino]-3-[l-[[3-[2-[(320487
    578
    P hydroxyphenyl)methylamino]ethoxy]-5,7-dimethyl-l-adamantyl]methyl]-5-methyl-pyrazol4-yl]pyridine-2-carboxylic acid, 2-[[6-(l,3-benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-methyl-amino]-5-[3-[4-[3(dimethy lamino)prop-1 -y nyl] -2-fluoro-phenoxy]propyl]thiazole-4-carboxylie acid, 2-[[6-(l,3-benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-(3-hydroxypropyl)amino]-5-[3[2-fluoro-4-[3-(methylamino)prop-l-ynyl]phenoxy]propyl]thiazole-4-carboxylic acid, 2-[[6-(l,3-benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-methyl-amino]-5-[3-[2-fluoro4-(3-pyrrolidin-l-ylprop-l-ynyl)phenoxy]propyl]thiazole-4-carboxylic acid, its enantiomers and diastereoisomers, and addition salts thereof with a pharmaceutically acceptable acid or base.
  22. 31. Pharmaceutical composition comprising a compound according to any of daims 1 to 30, or an addition sait thereof with a pharmaceutically acceptable acid or base in combination with one or more pharmaceutically acceptable excipients.
  23. 32. Pharmaceutical composition according to claim 31 for use as pro-apoptotic agents.
  24. 33. Pharmaceutical composition according to claim 31 for use in the treatment of cancers, autoimmune diseases or immune System diseases.
  25. 34. Pharmaceutical composition according to claim 33 wherein the cancer is an haematological malignancy or a solid tumor.
  26. 35. Pharmaceutical composition according to claim 34 wherein the haematological malignancy is myeloma, especially multiple myeloma, lymphoma, especially Non-Hodgkin Lymphoma (NHL), or leukemia, especially Chronic Lymphocytic Leukemia (CLL), T-cell Acute Lymphoblastic Leukemia (T-ALL), B-cell Acute Lymphoblastic Leukemia (B-ALL) and Acute Myelogenous Leukemia (AML).
  27. 36. Pharmaceutical composition according to claim 34 wherein the solid tumor is selected from bladder, brain, breast, utérus, œsophagus and liver cancers, colorectal cancer, rénal cancer, melanoma, ovarian cancer, prostate cancer, pancreatic cancer and lung cancer.
  28. 37. Compound according to any of daims 1 to 30, or an addition sait thereof with a
    579
    Pharmaceutically acceptable acid or base, for use in the treatment of a cancer selected from: myeloma, especially multiple myeloma, lymphoma, especially Non-Hodgkin Lymphoma (NHL), or leukemia, especially Chronic Lymphocytic Leukemia (CLL), T-cell Acute Lymphoblastic Leukemia (T-ALL), B-cell Acute Lymphoblastic Leukemia (B-ALL) and Acute Myelogenous Leukemia (AML), bladder, brain, breast, utérus, œsophagus and liver cancers, colorectal cancer, rénal cancer, melanoma, ovarian cancer, prostate cancer, pancreatic cancer and lung cancer, especially non-small-cell lung cancer and small-cell lung cancer.
  29. 38. Combination of a compound according to any of claims 1 to 30, with an anti-cancer agent selected from genotoxic agents, mitotic poisons, anti-metabolites, protéasome inhibitors, kinase inhibitors and antibodies.
  30. 39. Pharmaceutical composition comprising a combination according to claim 38 in combination with one or more pharmaceutically acceptable excipients.
  31. 40. Combination according to claim 38 for use in the treatment of cancers.
  32. 41. Compound according to any of claims 1 to 30, for use in the treatment of cancers requiring radio therapy.
  33. 42. Pharmaceutical composition according to claim 31 for use in the treatment of diseases or conditions characterized by an excess or a deregulated activity of platelets, especially pro-thrombotic conditions.
  34. 43. Synthesis intermediate which is:
    wherein R4 and m are as defined in general formula (I), preferably R4 represents a hydrogen atom.
OA1202200024 2019-07-29 2020-07-28 3,6-diamino-pyridazin-3-yl derivatives, pharmaceutical compositions containing them and their uses as pro-apoptotic agents. OA20487A (en)

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