WO2012040754A2 - Novel thiazolamine derivates as differentiation accelerators - Google Patents

Abstract

The present invention relates to the use of a thiazolamine derivative for acceleration of the differentiation of cells, characterized in that a 2-thiazolamine derivate of the general formula below is used: in which R₁ to R₃ are each independently selected from hydrogen, benzyl, aryl and heteroaryl or are absent, where not more than one of the R₁ to R₃ radicals is hydrogen, and R₄ is selected from benzyl, aryl and heteroaryl, where R₁ to R₄ are each independently optionally substituted by one or more substituents selected from halogen (F, Cl, Br, I), hydroxyl, C_1-3-alkyl, C_1-3-alkoxy, carboxyl, C_1-3-alkylcarbonyl, C_1-3-alkoxycarbonyl, nitro, amino, mono- and di-C_1-3-alkylamino and cyano, in which the alkyl groups are optionally halogenated; ^… (aa) in each case is a single bond or a double bond; n = 0 to 2, where, in the case that n = 1, the R₄ radical may be bonded to position 4 or 5 of the thiazole ring and, in the case that n = 2, the R₄ radicals are bonded to positions 4 and 5 of the thiazole ring.

Description

 New thiazolamine derivatives as differentiation accelerators

The present invention relates to novel thiazolamine derivatives for use as accelerators of cell differentiation.

Molecules containing a thiazolamine moiety exhibit a number of interesting biological activities, depending on their substituents and the substitution pattern on the thiazole ring. For example, compounds of the formula below have been shown to be potent inhibitors of histone deacetylase and tyrosine kinase:

wherein Ri and R _{2 are selected} from aryl, heteroaryl, hydrogen and acyl and R _{3 is selected} from aryl and heteroaryl (Mahboobi, S., et al., J. Med. Chem. 52 (8), 2265-2279 (2009)) , N-phenylthiazolamine derivatives are also known as selective aurora kinase inhibitors (CB Andersen et al., ACS Chemical Biology 3 (3), 180-192 (2008)). However, molecules of the formula below have been reported to induce autophagic cell death in renal cancer cells:

wherein Ri and R ₂ are selected from aryl and heteroaryl (MP Hay et al., J.Med.Chem. 53 (2), 787-797 (2010)). Due to the increasing importance of methods for influencing cell differentiation processes, attempts have been made to identify or synthesize molecules that have activity in promoting cell differentiation or undifferentiation. With regard to the use of thiazolamines for such purposes, N. Bouquier et al., Chem. Biol. 16, 657-666 (2009), have demonstrated that some substances, broadly referred to as thiazolamine derivatives, have the ability to Inhibition of the Trio / RhoG / Rac1 signaling pathway, inter alia, inhibits the differentiation of stimulated PC12 and C2C12 cells into neurites or myotubes. The thiazolamine derivatives used for this purpose are in all cases two- or three-membered fused ring systems in which a nitrogen of the thiazolamine represents a bridgehead atom, the following structure being predominant:

and in the other thiazolamine derivatives of the condensed benzene ring is missing. In the above formula, R stands for variously substituted aryls or heteroaryls.

WO 2005/047524 A2 discloses molecules of similar size based on purines which induce the dedifferentiation of lineage-determined mammalian cells ("lineage-committed mammalian cells"). Molecules in such size ranges, ie, molecular weights below about 1,000 g / mol, are often referred to as "small molecules", and the majority of all synthetic drugs are in this molecular weight range. In contrast to the above literature, in which differentiation inhibitors and de-differentiating agents are described, however, chemical compounds which promote the differentiation of cells, for example in the form of such "small molecules", are desirable for numerous applications, for example for stem cell therapy. Examples here- for are hardly known. For example, a compound called the adhesin has been reported to be able to accelerate the differentiation of hippocampal neurons (Hoshino, M. et al., Biochemical Journal 427, 297 (2010)).

 "Adhesamin"

And for derivatives of a compound known as wrenchnolol, which is reminiscent of a wrench, some groups of researchers have shown transcription and thus the expression of genes activating effects, among other things, inducing myogenesis of C2C12 cells (Jung, D. et al. J. Am. Chem. Soc. 131, 4774-4782 (2009)).

 "Wrenchnolol"

Both Wrenchnolol and Adhesamine are certainly complex compounds whose synthesis is associated with relatively high costs. Another interesting compound is neuropathiazole, a 4-thiazolamine that has been reported to induce the differentiation of multipotential neural progenitor cells of the hippocampus into neurons in adults.

 neuropathiazol

In WO 2007/064891 A1, new compounds are generally disclosed which are intended to induce the differentiation of mammalian cells into neuronal cells. The compounds correspond to the general formula

die folglich sowohl 2-, 4- als 5-Thiazolamine darstellen kann. Konkrete Synthesewege, basierend auf der altbekannten Thiazolsynthese nach Hantzsch, werden nur für 2- und 4-Thiazolamine angegeben. Tatsächlich hergestellt wurden ausschließlich 4-Thiazolamine, die auch in Differenzierungstests auf ihre Aktivität untersucht wurden. Am besten schnitt dabei das obige Neuropathiazol ab, wobei von insgesamt fünfzehn getesteten 4-Thiazolaminen vier unwirksam waren und zwei sogar antiproliferativ wirkten. Die Frage, ob 2- oder 5-Thiazolamine ähnliche Wirkung zeigen wür- den, bleibt offen.

which can thus represent both 2-, 4- as 5-thiazolamines. Concrete synthesis routes, based on the well-known thiazole synthesis according to Hantzsch, are given only for 2- and 4-thiazolamines. In fact, only 4-thiazolamines were prepared, which were also tested in differentiation tests for their activity. The best of these was the above neuropathiazole, of which four out of a total of fifteen tested 4-thiazolamines were inactive and two were even antiproliferative. The question of whether 2- or 5-thiazolamines would show similar effects remains open.

Therefore, the object of the present invention was to produce further small molecules, in particular those based on thiazolamine, which are capable of accelerating the differentiation of mammalian cells and which are relatively easy to synthesize. DISCLOSURE OF THE INVENTION

 This object is achieved by providing a group of novel 2-thiazolamine derivatives which conform to the following general formula:

R ₁ to R ₃ are independently selected or absent from hydrogen, benzyl, aryl and heteroaryl, with at most one of the radicals R ₁ to R _{3 being} hydrogen and R _{4 being selected} from benzyl, aryl and heteroaryl,

wherein R ₁ to R ₄ are each independently optionally substituted with one or more substituents selected from halogen (F, Cl, Br, I), hydroxy, Ci _-3 - alkyl, Ci _-3 alkoxy, carboxy, C _1-3 alkylcarbonyl , d-3-alkoxycarbonyl, nitro, amino, mono- and di-Ci-3-alkylamino and cyano, in which the alkyl groups are optionally halogenated substituted;

 each represents a single bond or a double bond; n = 0 to 2;

where, in the case n = 1, the radical R ₄ can be bonded to position 4 or 5 of the thiazole ring and, in the case n = 2, the radicals R ₄ are bonded to the positions 4 and 5 of the thiazole ring.

The above thiazolamine derivatives can be obtained in only three synthetic steps, whereby the cost and the cost of preparation compared to the compounds of the prior art can be significantly reduced. In addition, such derivatives have been shown to be effective in promoting cell differentiation in a C2C12 mouse cell model. Thus, the invention relates to the use of compounds of the above formula as cell differentiation accelerators.

Aryl and heteroaryl are to be understood here as mononuclear and polynuclear aromatic ring systems, preferably those having 5 to 10 ring atoms, more preferably aromatic rings having 5 or 6 ring atoms. The heteroatoms, of which one or more Others may be present, are preferably selected from nitrogen, sulfur and oxygen, with nitrogen and sulfur being particularly preferred and a single nitrogen heteroatom in the heteroaryl radical is more preferred. However, among the aryl and heteroaryl radicals, in particular phenyl is preferred.

Preferably, an optional double bond shown in broken line in the formula is present, either endo or exocyclic, ie extending from C2 to either the thiazole or amine nitrogen. Furthermore, R 1 to R _{3 are} preferably selected independently of one another from benzyl and phenyl, and on the two nitrogen atoms preferably only a total of two substituents are present. These two substituents are in particularly preferred embodiments each a phenyl and a benzyl group. The compounds according to the invention preferably have only one radical R ₄ , ie preferably: n = 1. This radical is particularly preferably bonded to C 5 of the thiazole ring and is preferably selected from aryl and heteroaryl, more preferably from phenyl, thiophenyl and pyridinyl wherein phenyl is particularly preferred.

The optional substituents from R 1 to R ₄ are preferably selected from fluoro, methyl, trifluoromethyl, nitro, carboxy and C 1-3 -alkoxycarbonyl, more preferably from fluoro, methyl and nitro. Without wishing to be bound by any particular theory, the inventors of the present application believe that, in the presence of larger or even bulky substituents, the interaction of the thiazolamine derivatives of the invention with translation factors or other components of the cells essential to induce cell differentiation is hindered the effect of the invention would be impaired. Furthermore, it is believed that more electron attractive substituents are to be preferred because they appear to exert an activating effect. Preferred thiazolamine derivatives of the invention are as follows

Compounds significant cell differentiation acceleration was observed without excessive toxicity:

 N-benzyl-N, 5-diphenylthiazolamine 23,

N-benzyl-N-phenyl-5- (2-fluoropyridin-3-yl) -thiazolamine 24,

 Ethyl 4- (2- (N-benzyl-N-phenylamino) thiazol-5-yl) benzoate 25,

 N-benzyl-N-phenyl-5- (m-nitrophenyl) -thiazolamine 28,

 N-Benzyl-N-phenyl-5- (p -fluorophenyl) -thiazolamine 31a.

 N-benzyl-N-phenyl-4,5-bis (p-fluorophenyl) -thiazolamine 31b,

N-benzyl-N-phenyl-5- (thiophen-2-yl) -thiazolamine 32,

 N- (3-Benzyl-5-phenylthiazol-2 (3H) -ylidene) aniline 34.

 N- (3-benzyl-5- (2-fluoropyridin-3-yl) thiazole-2 (3H) -ylidene) aniline 35,

 N- (3-Benzyl-5- (2-fluoropyridin-4-yl) thiazole-2 (3H) -ylidene) aniline 36,

 Ethyl 4- (3-benzyl-2- (phenylimino) -2,3-dihydrothiazol-5-yl) benzoate 37, N- (3-benzyl-5-o-tolylthiazol-2 (3H) -ylidene) aniline 38,

 N- (3-benzyl-5-p-tolylthiazol-2 (3H) -ylidene) aniline 39,

 N- (3-Benzyl-5- (4-nitrophenyl) thiazole-2 (3H) -ylidene) aniline 41,

 N- (3-Benzyl-5- (4-methoxyphenyl) thiazole-2 (3H) -ylidene) aniline 42,

 N- (3-Benzyl-5- (4-fluorophenyl) thiazole-2 (3H) -ylidene) aniline 43 and

N- (3-benzyl-5- (2-fluoropyridin-3-yl) thiazol-2 (3H) -ylidene) aniline-4-carboxylic acid ethyl ester

46th

Among them, due to total non-toxicity, the following compounds are even more preferred:

N-benzyl-N, 5-diphenylthiazolamine 23,

 N-benzyl-N-phenyl-5- (m-nitrophenyl) -thiazolamine 28,

 N- (3-Benzyl-5-phenylthiazol-2 (3H) -ylidene) aniline 34.

 N- (3-benzyl-5-p-tolylthiazol-2 (3H) -ylidene) aniline 39,

 N- (3-Benzyl-5- (4-methoxyphenyl) thiazole-2 (3H) -ylidene) aniline 42 and

N- (3-Benzyl-5- (4-fluorophenyl) thiazole-2 (3H) -ylidene) aniline 43,

of which the following three compounds give the best results in terms of

Acceleration of cell divergence have delivered: N-benzyl-N-phenyl-5- (m-nitrophenyl) -thiazolamine 28,

N- (3-Benzyl-5-p-tolylthiazol-2 (3H) -ylidene) aniline 39 and

N- (3-Benzyl-5- (4-fluorophenyl) thiazole-2 (3H) -ylidene) aniline 43. Hereinbelow, the present invention will be further illustrated by Examples and Comparative Examples, which are merely illustrative of preferred embodiments and are not intended to limit the invention in any way ,

EXAMPLES

A) Synthesis General

Melting points were determined on a Kofler hot stage microscope and are uncorrected. HR-MS analyzes were performed at the Institute for Chemical Technologies and Analytics at the Vienna University of Technology. When MS and MS / MS spectra were recorded, all samples were analyzed by LC-IT-TOF-MS, detection mode: positive ions. For the subsequent evaluation, only positive ionization spectra were used (where the quasi-molecular ion is the [M + H] ⁺ ion). Further data or observations were not considered. GC-MS measurements were taken on a Thermo Finnigan Focus GC / DSQ II using a standard capillary acid BGB 5 (30 mx 0.32 mm ID) and using one of the following standardized temperature profiles: STD 100-280 ° C Short method: 2 Minutes at 100 ° C, at 18 ° C / min to 280 ° C, 5 minutes at 280 ° C; or STD 100-280 ° C Long Procedure: 2 minutes at 100 ° C, at 6 ° C / min to 150 ° C, 3 minutes at 150 ° C, at 10 ° C / min to 280 ° C, 10 minutes at 280 ° C. Thin-layer chromatography was carried out with silica gel plates 60 F245 from Merck. Column chromatography was performed using Merck Kieselgel 60. NMR spectra were recorded on a BRUKER AC 200 FT NMR spectrometer with TMS (tetramethylsilane) as the internal standard. Solvents and reagents were obtained from commercial sources. The solvents were optionally distilled and dried before use. Synthetic Examples 1-9 - Preparation of Thiazolamines

General Procedure A

1.0 equivalents of the corresponding halothiazole, 1.5 equivalents of the corresponding amine and 0.5 equivalents of p-toluenesulfonic acid (p-TsOH) were dissolved in isopropanol (i-PrOH) and the reaction mixture was stirred at 80 ° C., until full turnover was observed. The reaction mixture was then diluted with ethyl acetate (EtOAc) and washed with saturated aqueous NaHCO ₃ solution and brine. The purification was carried out by Kugelrohr distillation. Synthetic Example 1

Preparation of N-phenyl-2-thiazolamine 3

 1 2 3

 According to general procedure A above, 41 mg (0.25 mmol) of 2-bromothiazole 1 were reacted with aniline 2 in the presence of p-TsOH. After stirring for 3 days and Kugelrohr distillation, the title compound 3 (40 mg, 90% of theory) was obtained.

Mp: 128-129 ° C (Lit .: 128-129 ° C)

R _f (PE / EtOAc 4: 1): 0.51

¹ H-NMR (200 MHz, CDCl ₃ ) δ: 6.63 (d, J = 3.6Hz, 1H, H5), 7.02-7.11 (m, J = 4.3Hz, 1H, H4 '), 7.30 (d, J = 3.6Hz, 1H, H4), 7.35-7.37 (m, 4H, PhH').

¹³ C-NMR (50 MHz, CDCl ₃ ) δ: 106.5 (d), 118.0 (d, C2 ', C6'), 122.5 (d), 129.3 (d, C3 \ C5 ' ), 138.0 (d), 141, 1 (s), 166.6 (s).

MS m / z (CI, NH ₃ ): 175 (100, M ⁺ ), 150 (16), 104 (19), 77 (27), 58 (18).

Synthetic Example 2

 Preparation of N- (4-morpholinophenyl) -2-thiazolamine 9

Gemäß dem obigen allgemeinen Verfahren A wurden 48 mg (0,40 mmol) 2-Chlor- thiazol 4 in Gegenwart von p-TsOH mit 4-Morpholinoanilin 5 umgesetzt. Nach 13 d Rühren und Kugelrohrdestillation wurde die Titelverbindung 9 (90 mg, 87 % d.Th.) als dunkelbrauner Feststoff erhalten.

According to general procedure A above, 48 mg (0.40 mmol) of 2-chlorothiazole 4 were reacted with 4-morpholinoaniline 5 in the presence of p-TsOH. After stirring for 13 days and Kugelrohr distillation, the title compound 9 (90 mg, 87% of theory) was obtained as a dark brown solid.

Mp .: 200-201 ° C

R _f (PE / EtOAc 2: 1): 0.19

¹ H-NMR (200 MHz, CDCl ₃ ) δ: 2.99-3.03 (m, 4H, -CH ₂ ), 3.70-3.75 (m, 4H, -CH ₂ ), 6.97 (d, J = 3.5Hz, 1H, H5), 6.90 (d, J = 8.8Hz, 2H, Η2 ', H6'), 7.17 (d, J = 3.6Hz, 1H , H4), 7.47 (d, J = 8.6Hz, 2H, Η3 ', H5'), 9.89 (s, 1H, NH).

¹³ C-NMR (50 MHz, CDCl ₃ ) δ: 49.2 (t, -CH ₂ ), 66.1 (t, -CH ₂ ), 107.4 (d), 116.0 (d, C2 ') , C6 '), 118.2 (d, C3 \ C5'), 134.1 (s), 138.8 (d), 145.9 (s), 164.7 (s).

MS m / z (Cl, NH _3): 261 (100, M ^+), 203 (58), 175 (10), 101 (16), 77 (8).

HRMS (Cl, NH _3): MH ^+, Found .: 262.1013; for C ₁₃ H ₁₅ N ₃ OS: 262, 1009.

Synthetic Example 3

Preparation of N- (4-methoxyphenyl) -2-thiazolamine 10

 4 6 10

According to general procedure A above, 60 mg (0.50 mmol) of 2-chlorothiazole 4 were reacted with 4-methoxyaniline 6 in the presence of p-TsOH. After stirring for 13 days and Kugelrohr distillation, the title compound 10 (100 mg, 97% of theory) was obtained as a dark brown solid. Mp: 126-127 ° C (Lit .: 126-127 ° C)

R _f (PE / EtOAc 2: 1): 0.65 H-NMR (200 MHz, CDCl ₃ ) δ: 3.81 (s, 3H, -OCH ₃ ), 6.55 (d, J = 3.6Hz , 1 H, H5), 6.91 (m, 2H, Η2 ', Η6'), 7.22 (d, J = 3.6Hz, 1H, H4), 7.23-7.31 (m, 2H, H3 *, H5 '). ¹³ C-NMR (50 MHz, DMSO-d ₆ ) δ: 55.0 (q, -OCH ₃ ), 107.4 (d), 1 14, 1 (d, C2 ', C6'), 1 18, 6 (d, C3 \ C5 '), 134.8 (d), 138.7 (s), 153.9 (s), 164, 1 (s).

MS m / z (Cl, NH _3): 206 (100, M ^+), 191 (92), 163 (10), 136 (18), 58 (8).

Synthesis Example 4

Preparation of N- (3-methoxyphenyl) -2-thiazolamine 1_1

 4 7 1 1

 According to general procedure A above, 60 mg (0.50 mmol) of 2-chlorothiazole 4 were reacted with 3-methoxyaniline 7 in the presence of p-TsOH. After 13 days of stirring and Kugelrohr distillation, recrystallization from LP and MPLC (LP: EtOAc 3: 1), the title compound JM (49 mg, 46% of theory) was obtained as a brown solid. Mp: 81-83 ° C

R _f (PE / EtOAc 2: 1): 0.85

¹ H-NMR (200 MHz, CDCl ₃ ) δ: 3.80 (s, 3H, -OCH ₃ ), 6.52 (d, J = 3.6Hz, 1H, H5), 6.90-6, 92 (m, 2H, PhH '), 7.20 (d, J = 3.6Hz, 1H, H4), 7.21 -7.30 (m, 2H, PhH').

¹³ C-NMR (50 MHz, DMSO-d ₆ ) δ: 54.4 (q, -OCH ₃ ), 102.0 (d, ArC), 105.9 (d, ArC), 108.4 (d, ArC), 109.3 (d, ArC), 129.5 (d, ArC), 138.3 (d, ArC), 142.9 (s), 159.5 (s), 163.8 (s) ,

MS m / z (CI, NH ₃ ): 206 (100), 190 (15), 163 (7), 103 (8). Synthetic Example 5

 Preparation of N- (2-methoxyphenyl) -2-thiazolamine

 4 8 12

According to general procedure A above, 60 mg (0.50 mmol) of 2-chlorothiazole 4 were reacted with 2-methoxyaniline 8 in the presence of p-TsOH. After stirring for 13 days and kugelrohr distillation, the title compound 12 (88 mg, 85% of theory) was obtained as a reddish brown solid.

Mp: 126-128 ° C

R _f (PE / EtOAc 2: 1): 0.85

¹ H-NMR (200 MHz, CDCl ₃ ) δ: 3.89 (s, 3H, -OCH ₃ ), 6.65 (d, J = 3.6Hz, 1H, H5), 6.87-7, 02 (m, 3H, PhH '), 7.31 (d, J = 3.6Hz, 1H, H4), 7.83 (bs, 1H, NH), 7.95-8.00 (m, 1H, PhH ').

MS m / z (CI, NH ₃ ): 206 (52, M ⁺ ), 175 (100), 147 (12), 108 (18), 78 (8).

Synthetic example 6

Preparation of 4- (thiazol-2-ylamino) benzoic acid ethyl ester 13

 4 17 13

According to general procedure A above, 60 mg (0.50 mmol) of 2-chlorothiazole 4 were reacted in the presence of p-TsOH with 4-aminobenzoic acid ethyl ester (enzocaine) 17. After stirring for 1.5 h and Kugelrohr distillation, the title compound 13 (120 mg, 97% of theory) was obtained as a tan solid.

Mp: 155-156 ° C

R _f (PE / EtOAc 2: 1): 0.75 ¹ H-NMR (200 MHz, CDCl ₃ ) δ: 1.40 (t, J = 7.1 Hz, 3H, -CH ₃ ), 4.37 (q, J = 7.1 Hz, 2H, -CH ₂ ), 6.76 (d, J = 3.6Hz, 1H, H5), 7.36-7.44 (m, 3H, Η2 ', Η6', H4), 8.04 (d, J = 8.8Hz, 2H, Η3 ', H5').

¹³ C-NMR (50 MHz, DMSO-d ₆ ) δ: 14.4 (q, -CH ₃ ), 60.8 (t, -OCH ₂ ), 108.7 (d, C5), 116.0 ( d, C2 ', C6'), 123.8 (s), 131, 4 (d, C3 \ C5 '), 138.3 (d), 144.5 (s), 164.3 (s), 166 , 2 (s, C = 0).

MS m / z (Cl, NH _3): 248 (100, M ^+), 202 (62), 175 (19), 131 (8), 102 (20), 58 (11).

Synthesis example 7

 Preparation of N- (2-chlorophenyl) -2-thiazolamine 14

 4 18 14

 According to general procedure A above, 60 mg (0.50 mmol) of 2-chlorothiazole 4 were reacted with 2-chloroaniline 18 in the presence of p-TsOH. After 2 d stirring and Kugelrohr distillation, the title compound 14 (100 mg, 95% of theory) was obtained as a colorless solid.

Mp .: 87-90 ° C (Lit .: 88-90 ° C)

R _f (PE / EtOAc 2: 1): 0.85

1H-NMR (200 MHz, CDCl3) δ: 6.61 (d, J = 3.6Hz, 1H, H5), 7.20-7.26 (m, 6H, PhH ', H4).

MS m / z (CI, NH ₃ ): 210 (100), 183 (12), 138 (17), 87 (18).

Synthetic Example 8

Preparation of N- (4-chlorophenyl) -2-thiazolamine 15

 4 19 15

According to general procedure A above, 60 mg (0.50 mmol) of 2-chlorothiazole 4 were reacted with 4-chloroaniline 19 in the presence of p-TsOH. After stirring for 2 h and kugelrohr distillation, the title compound 15 (80 mg, 76% of theory) was obtained as a colorless solid.

Mp .: 166-168 ° C (Lit .: 167-168 ° C)

1H-NMR (200 MHz, CDCl ₃ ) δ: 6.67 (d, J = 3.6Hz, H5), 7.26-7.32 (m, 6H, ArH, H4), 8.14 (s, 1H, -NH).

¹³ C-NMR (50 MHz, DMSO-d ₆₎ δ: 108.3 (d, ArC), 118.1 (d, ArC), 123.8 (d, ArC), 127.7 (d, ArC) , 136.4 (s), 138.0 (d, ArC), 140.5 (d, ArC), 162.9 (s), 163.5 (s).

MS m / z (CI, NH _3): 210 (32, M ^+), 175 (100), 131 (11), 87 (12).

Synthetic Example 9

Preparation of N- (4-nitrophenyl) -2-thiazolamine 16

 4 20 16

According to general procedure A above, 60 mg (0.50 mmol) of 2-chlorothiazole 4 were reacted with 4-nitroaniline 20 in the presence of p-TsOH. After stirring for 12 h and bulb tube distillation, the title compound 16 (104 mg, 98% of theory) was obtained as a pale orange solid.

Mp .: 164-165 ° C

1 H NMR (200 MHz, CDCl ₃ ) δ: 6.79 (d, J = 3.6Hz, H5), 7.40 (d, J = 3.6Hz, H4), 7.50 (t, J = 8.2Hz, 1H, H5 '), 7.74 (dd, J = 7.0Hz, J = 2.3Hz, 1H, H6'), 7.87 (dd, J = 7.1Hz, J = 2.0Hz, 1H, H4 '), 8.31 (s, 1H, -NH), 8.36-8.38 (m, 1H, H2'). ³ C-NMR (50 MHz, DMSO-d ₆ / acetone-d ₆₎ δ: 110.3 (d, ArC), 111, 8 (d, ArC), 116.3 (d, ArC), 123.5 (d, ArC), 130.7 (d, ArC), 140.0 (d, ArC), 143.4 (s), 149.8 (s), 164.2 (s). MS m / z (Cl, NH _3): 221 (100, M ^+), 174 (34), 131 (15), 90 (6), 58 (10).

HRMS (Cl, NH _3): MH ^+, Found .: 222.0332; calcd for C ₉ H 7 N ₃ O ₂ S: 222.0338.

Synthetic Examples 10 and 11 - Introduction of N-Benzyl Groups Synthesis Example 10

 Preparation of N-benzyl-N-phenyl-2-thiazolamine 21

 3 21

 N-phenyl-2-thiazolamine 3 (30 mg, 0.17 mmol, 1.0 equiv.), Benzylbromide (38 mg, 0.22 mmol, 1.3 equiv.) And NaH (4.1 mg, 0). 17 mmol, 1.0 equiv) were dissolved in dry DMF (2 ml) and stirred at room temperature for 5 h. The reaction mixture was then diluted with EtOAc, concentrated on silica gel on a rotary evaporator and purified by MPLC (LP: EtOAc 85:15 + 5% TEA). The title compound 21 was obtained as a beige solid (30 mg, 66% of theory).

Mp: 79-80 ° C

R _f (PE / EtOAc 5: 2): 0.20

1H-NMR (200 MHz, CDCl ₃ ) δ: 5.12 (s, 2H, -CH ₂ ), 6.40 (d, J = 3.7Hz, H 4), 7.15-7.33 (m, 10H, ArH).

3C-NMR (50 MHz, CDCI ₃₎ δ: 56.6 (t, CH _2), 107.7 (d), 126.2 (d, ArC), 126.8 (d, ArC),

127.2 (d, ArC), 127.7 (d, ArC); 128.5 (d, ArC), 129.8 (d, ArC), 137.7 (s), 139.3 (d),

145.3 (s), 170.9 (s).

MS m / z (CI, NH ₃ ): 266 (56, M ⁺ ), 174 (100), 167 (30), 131 (11), 91 (16).

HRMS (Cl, NH _3): MH \ Found .: 267.0955; Calc. for Ci ₆ H ₁₄ N ₂ S: 267.0950. Synthetic Example 11

 Preparation of N- (3-benzylthiazol-2 (3H) -ylidene) aniline 22

 3 22

N-phenyl-2-thiazolamine 3 (264 mg, 1.50 mmol, 1.0 equiv.), Benzyl bromide (334 mg, 1.96 mmol, 1.3 equiv.) And triethylamine (152 mg, 1.50 mmol , 1.0 equiv) were dissolved at 0 ° C in dry dioxane (4 ml), allowed to warm slowly and stirred at 120 ° C on a heating block for 3 days. The reaction mixture was then diluted with EtOAc, concentrated on silica gel on a rotary evaporator and purified by MPLC (LP: EtOAc 85:15 + 4% TEA). The title compound 22 was obtained as a pale yellow solid (287 mg, 72% of theory).

Mp .: 94-95 ° C

R _f (PE / EtOAc 5: 2): 0.20

¹ H-NMR (200 MHz, CDCl ₃ ) δ: 5.06 (s, 2H, -CH ₂ ), 5.85 (d, J = 4.9Hz, 1H, H5), 6.50 (d, J = 4.9Hz, 1H, H4), 7.00-7.10 (m, 3H, ArH), 7.33-7.37 (m, 7H, ArH).

¹³ C-NMR (50 MHz, CDCI ₃₎ δ: 49.8 (t, CH _2), 97.6 (d, 1 H, C5), 121, 4 (d, ArC), 123.2 (d ), 126.3 (s), 128.1 (s), 128.6 (d, ArC), 129.4 (d, ArC), 136.8 (s).

MS m / z (Cl, NH _3): 266 (42, M ^+), 167 (100), 131 (10), 91 (18), 15 (8).

HRMS (Cl, NH _3): MH \ Found .: 267.0954; calcd for C ₆ H ₁₄ N ₂ S: 267.0950.

Synthetic Example 12

 Preparation of N- (3-benzylthiazol-2 (3H) -ylidene) aniline 33

 13 33

Ethyl 4- (thiazol-2-ylamino) benzoate 13 (120 mg, 0.48 mmol, 1.0 equiv.), Benzyl bromide (106 mg, 0.62 mmol, 1.3 equiv.) And triethylamine (48 mg, 0 , 48 mmol, 1, 0 equiv.) Were dissolved at 0 ° C in dry dioxane (4 ml), allowed to warm slowly and stirred at 120 ° C on a heating block for 3 d. The reaction mixture was then diluted with EtOAc, concentrated on silica gel on a rotary evaporator and purified by MPLC (LP: EtOAc 85:15 + 4% TEA). The title compound 33 was obtained as a yellowish solid (138 mg, 85% of theory).

Examples 1 to 10 - Arylations of Thiazolamine 21

General procedure B

1.0 equivalents of thiazolamine derivative 21, 2.0 equivalents of the corresponding haloaryl, 2.0 equivalents of potassium acetate (KOAc) and 0.01 equivalents of Pd (OAc) ₂ were charged in this order into a reaction vessel. N, N-dimethylacetamide (DMAc) was added, after which the reaction vessel was evacuated and purged three times with argon. The reaction mixture was then stirred at 120 ° C until complete conversion was achieved. Thereafter, the reaction mixture was diluted with EtOAc and concentrated on silica gel directly on a rotary evaporator. Subsequent purification was by MPLC (unless otherwise indicated by LP: EtOAc 95: 5-0: 100 + 3% triethylamine). example 1

 Preparation of N-benzyl-N, 5-diphenylthiazolamine 23

 21 23

According to the above General Procedure B, 40 mg of 2J. reacted with iodobenzene for 48 h. Using MPLC, the title compound 23 was obtained as a tan solid (43 mg, 85% of theory).

 Mp .: 65-66 ° C

R _f (PE / EtOAc 5: 2): 0.45

1 H-NMR (200 MHz, CDCl ₃ ) δ: 5.12 (s, 2H, -CH ₂ ), 7.06-7.29 (m, 15H, ArH), 7.37 (s, 1H, H4) ,

3C-NMR (50 MHz, CDCI ₃₎ δ: 56.7 (t, CH _2), 125.4 (d, ArC), 126.4 (d, ArC), 126.7 (d, ArC), 127.1 (d, ArC), 127.3 (d, ArC), 127.9 (d, ArC), 128.5 (d, ArC), 128.8 (d, ArC), 129.9 (i.e. , ArC), 132.5 (s), 134.7 (d, Cl '"), 137.6 (s), 145.0 (s), 169.6 (s).

MS m / z (CI, NH ₃ ): 343 (16), 342 (62, M ⁺ ), 252 (20), 251 (100), 250 (79), 219 (19), 134 (20), 91 (73).

HRMS (CI, NH ₃ ): MH \ gef .: 343.1281; calcd for C22H18N2S: 343.1263.

Example 2

 Preparation of N-benzyl-N-phenyl-5- (2-fluoropyridin-3-yl) thiazolamine 24

 21 24

According to General Procedure B above, 60 mg of 2J. reacted with 2-fluoro-3-iodo-pyridine for 4 h. Using MPLC, the title compound 24 was obtained as a beige solid (78 mg, 94% of theory).

 Mp .: 117-118 ° C

R _f (PE / EtOAc 10: 1): 0.33

1 H NMR (200 MHz, CDCl ₃ ) δ: 5.13 (s, 2H, -CH ₂ ), 7.03 (dt, J = 5.4Hz, J _H F = 2.1HZ, 1H, H5 ^m ), 7.18-7.32 (m, 10H, ArH), 7.59-7.64 (m, 2H, ArH), 7.90 (d, J = 3.6Hz, ArH).

¹³ C-NMR (50 MHz, CDCl ₃ ) δ: 55.9 (t, -CH ₂ ), 116.1 (dd, J = 29.2Hz, C6 ^{, M} ), 118.8 (ds, J = 9 , 1 Hz, C1 ^{, M} ), 121, 7 (ds, J _C F = 4,3HZ, C5), 126,7 (d, ArC), 127,5 (d, ArC), 127,6 (d, ArC), 127.9 (d, ArC), 128.5 (d, ArC), 130.1 (d, ArC), 137.1 (dd, J = 4.8Hz, C5 '''), 137.2 (s, Ar), 139.8 (dd, J _C F = 10.0HZ, C4), 144.3 (dd, J _C F = 14.7HZ, C4 '"), 144.6 (s, Ar) , 158.6 (ds, J _C F = 241 HZ, C2 ^{, m} ), 170.9 (s).

MS m / z (CI, NH _3): 361 (45, M ^+), 269 (52), 167 (64), 140 (10), 110 (12), 91 (100). HRMS (Cl, NH _3): MH ^+, Found .: 362.1130; calcd for C ₂ iHi ₆ FN ₃ S: 362.1122.

Example 3

 Preparation of 4- (2- (N-benzyl-N-phenylamino) thiazol-5-yl) -benzoic acid ethyl ester 25

 21 25

According to General Procedure B above, 32 mg of 2Λ were reacted with 4-iodobenzoic acid ethyl ester for 24 hours. Using MPLC, the title compound 25 was obtained as a yellow oil (30 mg, 60% of theory).

R _f (PE / EtOAc 10: 1): 0.48

H-NMR (200 MHz, CDCl ₃ ) δ: 1.28 (t, J = 7.1 Hz, 3H, -CH ₃ ), 4.24 (q, J = 7, 1 Hz, 2H, -OCH ₂ ), 5.10 (s, 2H, CH _2), 7, 14 to 7.32 (m, 13H, ArH), 7.47 (s, 1 H, H4), 7.84 (d, J = . 8.4 Hz, H3 "\ ^{H5, M)} 3C-NMR (50 MHz, CDCI ₃₎ δ: 14.1 (q, -CH _3), 55.8 (t, CH _2), 61, 2 ( t, -OCH ₂ ), 124.6 (d, ArC), 126.4 (s), 126.6 (d, ArC), 127.4 (d, ArC), 127.5 (d, ArC), 127.8 (d, ArC), 128.1 (s), 128.5 (d, ArC), 130.0 (d, ArC), 130.2 (d, ArC), 136.8 (d, ArC ), 137.4 (s), 144.6 (s), 166.6 (s), 170.4 (s, C = O) * (* one singlet coincides with another peak).

HRMS (Cl, NH _3): MH ^+, Found .: 415.1478; calcd for C ₂₅ H ₂₂ N ₂ O ₂ S: 414, 1475.

Example 4 - Comparative Example

 Preparation of N-benzyl-N-phenyl-5- (o-tolyl) thiazolam

 21 26

According to General Procedure B above, 40 mg of 2Λ were reacted with 2-iodotoluene for 20 hours. Using MPLC, the title compound 26 was obtained as a beige solid (43 mg, 80% of theory).

Mp .: 71-73 ° C

R _f (PE / EtOAc 10: 1): 0.44

1H-NMR (200 MHz, CDCl ₃ ) δ: 2.31 (s, 3H, -CH ₃ ), 5.12 (s, 2H, -CH ₂ ), 7.1 1 -7.28 (m,

15H, ArH).

C-NMR (50 MHz, CDCl ₃ ) δ: 21, 4 (q, -CH ₃ ), 56.2 (t, -CH ₂ ), 126.0 (d, ArC), 126.1 (s), 126.3 (d, ArC), 126.9 (d, ArC), 127.3 (d, ArC), 127.4 (d, ArC), 127.8 (d, ArC), 128.5 (i.e. , ArC), 129.8 (d, ArC), 130.2 (d, ArC), 130.8 (d, ArC), 131, 4 (s), 136.0 (s), 137.4 (i.e. , ArC), 137.6 (s), 145, 1 (s), 170.1 (s).

MS m / z (Cl, NH _3): 356 (65, M ^+), 167 (88), 147 (38), 1 15 (88), 91 (100), 65 (26).

HRMS (Cl, NH _3): MH ^+, Found .: 357.1427; calc. for C ₂₃ H2oN ₂ S: 357, 1420.

Example 5 - Comparative Example

 Preparation of N-benzyl-N-phenyl-5- (p-tolyl) thiazolamine 27

 21 27

According to the above General Procedure B, 75 mg of 2 ± was reacted with 4-iodotoluene for 20 hours. The title compound 27 was obtained as a beige solid by MPLC (77 mg, 77% of theory).

 Mp: 105-108 ° C

R _f (PE / EtOAc 10: 1): 0.48

1 H-NMR (200 MHz, CDCl ₃ ) δ: 2.22 (s, 3H, -CH ₃ ), 5.1 1 (s, 2H, -CH ₂ ), 6.98-7.02 (m, 3H , ArH), 7, 15-7.26 (m, 12H, ArH), 7.32 (s, H, H4).

¹³ C-NMR (50 MHz, CDCl ₃ ) δ: 21, 2 (q, -CH ₃ ), 56.1 (t, -CH ₂ ), 125.4 (d, ArC), 126.3 (d, ArC), 127.0 (d, ArC), 127.3 (d, ArC), 127.8 (d, ArC), 127.9 (s), 128.5 (d, ArC), 129.5 ( d, ArC), 129.9 (d, ArC), 134.3 (d, ArC), 136.5 (s), 137.6 (s), 145.0 (s), 169.3 (s) ^* (* one singlet coincides with another peak).

MS m / z (Cl, NH _3): 356 (48, M ^+), 265 (100), 233 (10), 167 (44), 105 (10), 91 (58).

HRMS (Cl, NH _3): MH \ Found .: 357.1427; calcd for C ₂₃ H ₂ oN ₂ S: 357.1420.

Example 6

 Preparation of N-benzyl-N-phenyl-5- (m-nitrophenyl) thiazole amine 28

 21 28

According to the above General Procedure B, 30 mg of 2J _. reacted with 3-iodonitrobenzene for 20 h. MPLC (LP: EtOAc 75: 25 → -0: 100 + 3% triethylamine) gave the title compound 28 as a light orange solid (41 mg, 51% of theory).

 Mp .: 109-110 ° C

R _f (PE / EtOAc 5: 2): 0.69

H-NMR (200 MHz, DMSO-d ₆ ) δ: 5.23 (s, 2H, -CH ₂ ), 7.24-7.35 (m, 6H, ArH), 7.45-7.50 ( m, 3H, ArH), 7.58 (t, J = 8.0Hz, 1H, H5 ^{, M} ), 7.92 (s, 1H, H4), 8.00 (dd, J = 8.0Hz , J = 2.0 Hz, 1 H, H4 ^m), 8.16 (s, 1 H, ^{H2, m),} 8.28 (dt, J = 7.3 Hz, J = 2.1 Hz, 1 H, H4 ^{, M} ), 8.84 (d, J = 8.0Hz, 1H, ArH).

1 ³ C-NMR (50 MHz, DMSO-d ₆ ) δ: 55.2 (t, -CH ₂ ), 118.6 (d, ArC), 120.8 (d, ArC), 123.2 (s ), 123.9 (d, ArC), 127.2 (d, ArC), 127.3 (d, ArC), 127.4 (d, ArC), 128.4 (d, ArC), 130.0 (d, ArC), 130.5 (d, ArC), 130.9 (d, ArC), 133.5 (d, ArC), 133.7 (s), 137.3 (d, ArC), 138 , 1 (s), 144.1 (s), 148.3 (s), 169.7 (s).

HRMS (Cl, NH _3): MH ^+, Found .: 388.1118; calcd for C22H17N3O2S: 388.1114.

Example 7 - Comparative Example

 Preparation of N-benzyl-N-phenyl-5- (p-nitrophenyl) -thiazolamine 29

 21 29

According to General Procedure B above, 69 mg of 2_1 were reacted with 4-iodonitrobenzene for 20 hours. MPLC (LP: EtOAc 70:30 -0: 100 + 3% triethylamine) gave the title compound 29 as a dark orange solid (20 mg, 20% of theory).

 Mp: 197-200 ° C

R _f (PE / EtOAc 5: 2): 0.66

¹ H-NMR (200 MHz, CDCl ₃ ) δ: 5.25 (s, 2H, -CH ₂ ), 7.30-7.47 (m, 6H, ArH), 7.46-7.48 (m , 4H, ArH), 7.66 (d, J = 8.8Hz, 2H, H2 ^{, M} , H4 "'), 8.00 (s, 1H, H4), 8.13 (d, J = 8 , 7Hz, Η3 '", H5'").

3C-NMR (50 MHz, acetone-d ₆ ) δ: 55.9 (t, -CH ₂ ), 124.3 (d, ArC), 124.8 (s), 125.0 (d, ArC), 126.9 (d, ArC), 127.3 (d, ArC), 127.8 (d, ArC), 128.0 (d, ArC), 128.4 (d, ArC), 130.1 (i.e. , ArC), 137.6 (s), 139.3 (s), 139.4 (d, ArC), 144.5 (s), 145.6 (s), 171, 4 (s).

HRMS (Cl, NH _3): MH ^+, Found .: 388.1118; Calcd for _{C22 H17} N30 ₂ S:. 388.1114.

Example 8

 Preparation of N-benzyl-N-phenyl-5- (p-methoxyphenyl) thiazolamine 30

 21 30

According to the above General Procedure B, 30 mg of 2 ± was reacted with 4-iodoanisole for 20 hours. MPLC gave the title compound 30 as a beige oil (21 mg, 47% of theory).

R _f (PE / EtOAc 5: 2): 0.66

H-NMR (200 MHz, CDCl ₃ ) δ: 3.72 (s, 3H, -OCH ₃ ), 5.12 (s, 2H, -CH ₂ ), 6.74-6.79 (m, 2H, Ar H).

3C-NMR (50 MHz, CDCl ₃ ) δ: 54.5 (q, -OCH ₃ ), 56.4 (t, -CH ₂ ), 1 14.3 (d, ArC), 125.1 (s) , 126.2 (d, ArC), 126.8 (d, ArC), 126.9 (d, ArC), 127.2 (d, ArC), 127.6 (s), 127.7 (d, ArC), 128.5 (d, ArC), 129.8 (d, ArC), 133.6 (d, ArC), 137.5 (s), 145.1 (s), 158.6 (s) , 169.0 (s).

HRMS (Cl, NH _3): MH ^+, Found .: 373.1374; calc. for C ₂₃ H ₂ O ₂ OS: 373.1369.

Example 9

 Preparation of N-benzyl-N-phenyl-5- (p -fluorophenyl) thiazolamine 31a and

N-benzyl-N-phenyl-4,5-bis (p-fluorophenyl) thiazolamine 31b

According to the above General Procedure B, 40 mg of 2J _. reacted with 4-iodofluorobenzene for 20 h. MPLC gave the title compound 31a as a brown solid (39 mg, 71% of theory) and the title compound 31b as a yellow solid (8 mg, 12% of theory).

31a:

R _f (PE / EtOAc 10: 1): 0.40

¹ H-NMR (200 MHz, CDCl ₃ ) δ: 5.05 (s, 2H, -CH ₂ ), 6.85-6.95 (m, 2H, ArH), 7.17-7.30 (m , 13H, ArH).

¹³ C-NMR (50 MHz, CDCl ₃ ) δ: 55.5 (t, -CH ₂ ), 114.6 (dd, J _C F = 22HZ, C2 "\ C6 ^* "), 125.3 (d, ArC), 125.5 (s, Ar), 125.9 (dd, J _C F = 8HZ, C3 ^m , C6 '''), 126.1 (d, ArC), 126.3 (d, ArC), 126.7 (d, ArC), 127.5 (d, ArC), 127.6 (s, Ar), 128.9 (d, ArC), 133.7 (d, ArC), 136.4 (s , Ar), 144.0 (s, Ar), 160.1 (ds, J _CF = 247Hz, C4 ^{, M} ), 168.5 (s).

MS m / z (CI, NH ₃ ): 360 (49, M ⁺ ), 269 (100), 91 (94), 167 (72), 152 (24).

HRMS (Cl, NH _3): MH \ Found .: 361, 1171; calcd for C22H17FN2S: 362.1169. 31b:

¹ H-NMR (200 MHz, CDCl ₃ ) δ: 5.24 (s, 2H, -CH ₂ ), 6.87-7.02 (m, 4H, ArH), 7.14-7.52 (m , 14H, ArH).

¹³ C-NMR (50 MHz, CDCl ₃ ) δ: 55.9 (t, -CH ₂ ), 115.1 (d, J _CF = 29Hz), 115.5 (d, J _CF = 29Hz), 126, 7 (d), 127.2 (d), 127.3 (d), 128.3 (d), 128.4 (d), 129.9 (d, ArC), 130.8 (d, J _CF = 28Hz), 130.9 (d, J _CF = 28Hz).

Example 10

 Preparation of N-benzyl-N-phenyl-5- (thiophen-2-yl) thiazolamine 32

 21 32

According to General Procedure B above, 40 mg of 21 were reacted with 2-iodothiophene for 20 hours. Using MPLC, the title compound 32 was obtained as a brown oil (41 mg, 82% of theory).

R _f (PE / EtOAc 10: 1): 0.40

¹ H-NMR (200 MHz, CDCl ₃ ) δ: 5.19 (s, 2H, -CH ₂ ), 6.90-6.96 (m, 2H, ArH), 7.10-7.11 (dd , J = 4.0Hz, J = 1, 2Hz, 1H, H3 ^{, M} ), 7.13-7.44 (m, 11H, ArH).

¹³ C-NMR (50 MHz, CDCI ₃₎ δ: 56.2 (t, CH _2), 120.9 (s), 123.4 (d, ArC), 126.4 (d, ArC), 127 , 3 (d, ArC), 127.5 (d, ArC), 127.8 (d, ArC), 128.5 (d, ArC), 129.9 (d, ArC), 134.6 (s) , 135.4 (d, ArC), 137.4 (s), 144.8 (s), 169.5 (s).

MS m / z (CI, NH ₃ ): 348 (89, M ⁺ ), 257 (100), 207 (48), 167 (40), 225 (34), 91 (10).

HRMS (Cl, NH _3): MH ^+, Found .: 349.0832; calc. for C ₂₀ Hi ₆ N ₂ S ₂ : 349.0828. Examples 11 to 21 - Arylations of Thiazolimine 22

 For these arylations, the above General Method B was also used essentially. Example 11

 Preparation of N- (3-benzyl-5-phenylthiazol-2 (3H) -ylidene) aniline 34

22 34

 According to the above General Procedure B, 100 mg of 22 was reacted with iodobenzene for 48 hours. Using MPLC, the title compound 34 was obtained as a tan solid (99 mg, 79% of theory).

 Mp .: 65-66 ° C

R _f (PE / EtOAc 5: 2): 0.52

¹ H-NMR (200 MHz, CDCl ₃ ) δ: 5.02 (s, 1 H, -CH ₂ ), 6.71 (s, 1 H, H 4), 6.95-7.29 (m, 15H , ArH).

¹³ C-NMR (50 MHz, CDCl ₃ ) δ: 50.2 (t, -CH ₂ ), 115.3 (s), 121, 7 (d, ArC), 121, 7 (d), 123.1 (d, ArC), 124.4 (d, ArC), 127.0 (d, ArC), 128.0 (d, ArC), 128.8 (d, ArC), 129.3 (d, ArC) , 131, 2 (s), 136,2 (s), 151, 1 (s) * ( ^* one singlet coincides with another peak).

MS m / z (Cl, NH _3): 342 (36, M ^+), 281 (18), 251 (32), 207 (68), 167 (100), 91 (10). HRMS (Cl, NH _3): MH ^+, Found .: 343.1271; Calc. for C ₂ 2Hi ₈ N ₂ S: 343.1263. Example 12

 Preparation of N- (3-Benzyl-5- (2-fluoropyridin-3-yl) thiazole-2 (3H) -ylidene) aniline 35

22 35

According to General Procedure B above, 70 mg of 22 was reacted with 2-fluoro-3-iodo-pyridine for 20 hours. MPLC (LP: EtOAc 80:20 -> - 0: 100 + 3% triethylamine) gave the title compound 35 as a tan solid (48 mg, 51% of theory).

 Mp: 139-140 ° C

R _f (PE / EtOAc 10: 2): 0.33

¹ H-NMR (200 MHz, CDCl ₃ ) δ: 7.06-7.13 (m, 4H, ArH), 7.24 (ds, J _H F = 1, 0HZ, 1 H, H4), 7, 34-7.49 (m, 8H, ArH).

¹³ C-NMR (50 MHz, CDCl ₃ ): 49.8 (t, -CH ₂ ), 106.9 (dd, J _C F = 8.5HZ, C4), 1.15.6 (dd, J _CF = 28.0Hz, C6 ^{, M} ), 121, 5 (d, ArC), 121, 8 (dd, J _C F = 4.3HZ, C5 '"), 123.4 (d, ArC), 127.8 ( dd, J _CF = 15.9Hz, ArC, C4 ^{, M} ), 127.9 (s, Ar), 128.0 (d, ArC), 128.9 (d, ArC), 129.6 (d, ArC ), 136.1 (s, Ar), 136.3 (s, Ar), 137.2 (d, ArC), 143.7 (ds, J _C F = 15.2 Hz, CT "), 151, 1 (s, Ar), 156.2 (s), 158.7 (ds, J = 240Hz, C2 ^m).

MS m / z (Cl, NH _3): 361 (26, M ^+), 269 (30), 167 (100), 1 10 (8), 91 (75).

HRMS (Cl, NH _3): MH ^+, 362.1 Found .: 131; calcd for C ₂ iH ₁₆ FN ₃ S: 362.1122. Example 13

 Preparation of N- (3-Benzyl-5- (2-fluoropyridin-4-yl) thiazole-2 (3H) -ylidene) aniline 36

22 36

According to General Procedure B above, 60 mg of 22 were reacted with 2-fluoro-4-iodo-pyridine for 10 hours. MPLC (LP: EtOAc 80: 20 → 0: 100 + 3% triethylamine) gave the title compound 36 as a tan solid (80 mg, 96% of theory).

 Mp: 111-112 ° C

R _f (PE / EtOAc 10: 2): 0.33

H-NMR (200 MHz, CDCI ₃₎ δ: 5.17 (s, 2H, CH _2), 6.73 (s, 1 H, H4), 7.06 (td, J = 5.4 Hz, J HF = 1, 7Hz, 1 H, H6 ^m), 7.21 to 7.42 (m, 10H, ArH), 7.67 (s, 1 H, H4), 8.00 (d, J = 8.0Hz , 1 H, H5 '").

¹³ C-NMR (50 MHz, CDCl ₃ ) δ: 56.3 (t, -CH ₂ ), 104.0 (dd, J _CF = 39.4Hz, C5 ^{, M} ), 117.0 (dd, JC _F = 3.7HZ, C6 '"), 123.4 (ds, J _C F = 4.1 HZ, C5), 126.8 (d, ArC), 127.8 (dd, J _C F = 22HZ, C2 '"), 127.9 (d, ArC), 128.6 (d, ArC), 138.2 (d, ArC), 136.8 (s, Ar), 139.4 (d, ArC), 144 , 3 (s, Ar), 145.2 (ds, J _CF = 8.9Hz, CT '), 147.8 (dd, J _C F = 16.1 HZ, C5 ^{, m} ), 164.5 (ds , J _C F = 250HZ, C3 '"), 171, 6 (s).

MS m / z (CI, NH ₃ ): 361 (58, M ⁺ ), 281 (28), 269 (100), 253 (26), 207 (76), 167 (42), 91 (14).

HRMS (Cl, NH _3): MH ^+, Found .: 362.1129; calcd for C ₂ iHi ₆ FN ₃ S: 362.1122. Example 14

 Preparation of 4- (3-Benzyl-2- (phenylimino) -2,3-dihydrothiazol-5-yl) benzoic acid ethyl ester 37

22 37

 According to the above General Procedure B, 30 mg of 22 was reacted with iodobenzene for 24 hours. Using MPLC, the title compound 37 was obtained as a yellow oil (29 mg, 58% of theory).

R _f (PE / EtOAc 10: 1): 0.51

1 H NMR (200 MHz, CDCl ₃ ) δ: 1, 38 (t, J = 7.1 Hz, -CH ₃ ), 4.37 (q, J = 7.1Hz, 2H, -OCH ₂ ), 5 , 14 (s, 2H, -CH ₂ ), 7.00 (s, 1 H, H4), 7.12 (d, J = 8.5Hz, 2H, H2 "\ H6 ^{, M} ), 7.24- 7.28 (m, 3H, ArH), 7.37-7.40 (m, 7H, ArH), 7.94 (d, J = 8.5 Hz, 2H, H3 "\ H5 ^m).

¹³ C-NMR (50 MHz, CDCl ₃ ) δ: 14.5 (q, CH ₃ ), 49.6 (t, -OCH ₂ ), 60.3 (t, -CH ₂ ), 113.6 (i.e. ), 121, 4 (d, ArC), 123.4 (d, ArC), 123.7 (s), 123.8 (d, ArC), 128.0 (d, ArC), 128.1 (s , Ar), 128.9 (d, ArC), 129.5 (d, ArC), 130.1 (d, ArC), 136.0 (s), 136.4 (s), 151, 2 (s ), 156.9 (s), 166.1 (s, C = 0).

HRMS (Cl, NH _3): MH \ Found .: 415.1475; calcd for C ₂₅ H ₂₂ N ₂ O ₂ S: 415.1485.

Example 15

 Preparation of N- (3-Benzyl-5- (o-tolyl) thiazole-2 (3H) -ylidene) aniline 38

22 38

According to General Procedure B above, 60 mg of 22 were reacted with 2-iodotoluene for 12 hours. Using MPLC, the title compound 38 was obtained as a brown oil (60 mg, 73% of theory).

R _f (PE / EtOAc 4: 1): 0.40

¹ H-NMR (200 MHz, CDCl ₃ ) δ: 2.25 (s, 3H, -CH ₃ ), 5.04 (s, 2H, -CH ₂ ), 6.42 (s, 1 H, H 4) , 6.93-7.30 (m, 15H, ArH).

¹³ C-NMR (50 MHz, CDCl ₃ ) δ: 21, 1 (q, -CH ₃ ), 49.7 (t, -CH ₂ ), 115.2 (s), 121, 1 (d, ArC) , 121, 5 (d, ArC), 123.1 (d, ArC), 124.4 (d, ArC), 127.9 (d, ArC), 128.0 (d, ArC), 128.8 ( d, ArC), 128.9 (s), 129.4 (d, ArC), 136.7 (s), 136.9 (s), 151, 5 (s), 157.5 (s).

MS m / z (Cl, NH _3): 356 (50, M ^+), 281 (6), 265 (26), 207 (21), 115 (20), 167 (100). HRMS (Cl, NH _3): MH \ Found .: 357.1428; calc. for C ₂₃ H2oN ₂ S: 357.1420.

Example 16

 Preparation of N- (3-Benzyl-5- (p-tolyl) thiazole-2 (3H) -ylidene) aniline 39

22 39

According to General Method B above, 60 mg of 22 was reacted with 4-iodotoluene for 18 hours. Using MPLC, the title compound 38 was obtained as a brown solid (56 mg, 68% of theory).

 Mp .: 59-60 ° C

R _f (PE / EtOAc 4: 1): 0.41

H-NMR (200 MHz, CDCI ₃₎ δ: 2.49 (s, 3H, _-CH3), 5.26 (s, 2H, CH _2), 6.90 (s, 1 H, H4), 7.20-7.29 (m, 6H, ArH), 7.40-7.55 (m, 8H, ArH).

¹³ C-NMR (50 MHz, CDCl ₃ ) δ: 21, 7 (q, -CH ₃ ), 50.3 (t, -CH ₂ ), 114.5 (s), 122.0 (d, ArC) , 123.5 (d, ArC), 124.7 (d, ArC), 126.6 (d, ArC), 128.2 (d, ArC), 128.3 (d, ArC), 128.6 ( d, ArC), 129.3 (d, ArC), 129.9 (d, ArC), 131, 4 (d, ArC), 131, 5 (s), 136.4 (s), 137.0 ( s), 151, 8 (s), 158.5 (s).

MS m / z (Cl, NH _3): 356 (90, M ^+), 281 (12), 264 (100), 233 (16), 207 (26), 167 (20), 91 (6) -

HRMS (Cl, NH _3): MH ^+, Found .: 357.1426; calcd for C ₂₃ H ₂ oN ₂ S: 357.1420.

Example 17

 Preparation of N- (3-Benzyl-5- (3-nitrophenyl) thiazole-2 (3H) -ylidene) aniline 40

According to General Procedure B above, 30 mg of 22 was reacted with 3-iodonitrobenzene for 3 hours. MPLC (LP: EtOAc 80: 20 → -0: 100 + 3% triethylamine) gave the title compound 40 as an orange solid (40 mg, 87% of theory).

Mp .: 134-146 ° C

R _f (PE / EtOAc 5: 2): 0.68

1H-NMR (200 MHz, DMSO-d ₆₎ δ: 6.99-7.04 (m, 3H, Ar H, H4), 7.31-7.45 (m, 7H, ArH), 7,49- 7.75 (m, 2H, ArH), 8.00 (d, J = 8.7Hz, 1H, ArH), 8.15-8.23 (m, 2H, H3 "\ H5 '").

¹³ C-NMR (50 MHz, DMSO-d ₆ ) δ: 49.4 (t, -CH ₂ ), 110.5 (s), 117.9 (d, ArC), 120.9 (d, ArC) , 123.2 (d, ArC), 127.6 (d, ArC), 127.8 (d, ArC), 128.6 (d, ArC), 129.5 (d, ArC), 130.0 ( d, ArC), 130.5 (d, ArC), 133.1 (s), 136.9 (s), 148.1 (s), 150.4 (s), 155.4 (s). HRMS (Cl, NH _3): MH ^+, Found .: 388.1118; calc. for C ₂₂ Hi7N ₃ O ₂ S: 388,1114.

Example 18

Preparation of N- (3-Benzyl-5- (4-nitrophenyl) thiazole-2 (3H) -ylidene) aniline 4J _.

22 41

According to the above General Procedure B, 23 mg of 22 were reacted with 4-iodonitrobenzene for 2 hours. MPLC (LP: EtOAc 80: 20 ^ -0: 100 + 3% triethylamine) gave the title compound 4J. as a red oil (30 mg, 86% of theory).

R _f (PE / EtOAc 4: 1): 0.40

¹ H-NMR (200 MHz, DMSO-d ₆ ) δ: 5.14 (s, 2H, -CH ₂ ), 7.00 (d, J = 7.4Hz, 2H, ArH), 7.09-7 , 39 (m, 8H, ArH), 7.57 (d, 2H), 8.15 (d, J = 8.8Hz, H3 "\ H5 '"), 8.26 (s, 1H, H4) , 3C-NMR (50 MHz, acetone-d ₆ ) δ: 50.5 (t, -CH ₂ ), 112.6 (s), 121, 9 (d, ArC), 124.2 (d, ArC), 125.0 (s), 125.1 (d, ArC), 125.2 (d, ArC), 128.7 (d, ArC), 129.0 (d, ArC), 129.1 (d, ArC ), 129.6 (d, ArC), 130.3 (d, ArC), 138.0 (s), 139.4 (s), 146.5 (s), 151, 9 (s).

HRMS (Cl, NH _3): MH \ Found .: 388.1118; calcd for C22H17N3O2S: 388.1114.

Example 19

 Preparation of N- (3-Benzyl-5- (4-methoxyphenyl) thiazole-2 (3H) -ylidene) aniline 42

22 42

According to General Procedure B above, 30 mg of 22 was reacted with 4-iodoanisole for 18 hours. MPLC (LP: EtOAc 12: 1- ^ 0: 100 + 3% triethylamine) gave the title compound 42 as a brown oil (19 mg, 42% of theory).

R _f (PE / EtOAc 4: 1): 0.40

¹ H-NMR (200 MHz, DMSO-d ₆ ) δ: 3.74 (s, 3H, -OCH ₃ ), 5.21 (s, 2H, -CH ₂ ), 6.89 (d, J = 8 , 7Hz, 2H, ArH), 7.29-7.49 (m, 13H, ArH).

¹³ C-NMR (50 MHz, CDCl ₃ ) δ: 55.1 (q, -OCH ₃ ), 55.8 (t, -CH ₂ ), 114.3 (d, ArC), 125.1 (s) , 126.2 (d, ArC), 126.8 (d, ArC), 126.9 (d, ArC), 127.2 (d, ArC), 127.7 (d, ArC), 128.4 ( d, ArC), 129.8 (d, ArC), 133.6 (d, ArC), 137.6 (s), 145.1 (s), 158.6 (s), 168.8 (s) ,

HRMS (Cl, NH _3): MH ^+, Found .: 373.1373; calc. for C ₂₃ H ₂ O ₂ OS: 373.1369.

Example 20

 Preparation of N- (3-benzyl-5- (4-fluorophenyl) thiazole-2 (3H) -ylidene) aniline 43

22 43

According to General Procedure B above, 30 mg of 22 was reacted with 4-iodofluorobenzene for 24 hours. MPLC (LP: EtOAc 12: 1 → -0: 100 + 3% triethylamine) gave the title compound 43 as a brown oil (19 mg, 46% of theory).

R _f (PE / EtOAc 10: 1): 0.48

H-NMR (200 MHz, CDCI ₃₎ δ: 5.12 (s, 2H, CH _2), 6.73 (s, 1 H, H4), 6.93 to 7.22 (m, 7H, ArH ), 7.33-7.39 (m, 7H, ArH).

3C-NMR (50 MHz, CDCI ₃₎ δ: 50.48 (t, CH _2), 1 15.7 (dd, J = 22,0Hz, C2 ^m, C6 ^m), 121 4 (d, ArC ), 123.2 (d, ArC), 126.2 (dd, J _C F = 8.0HZ, C3 "', C5'"), 127.3 (d, ArC), 128.0 (d, ArC ), 128.1 (d, ArC), 128.5 (d, ArC), 128.7 (d, ArC), 129.5 (d, ArC), 129.9 (d, ArC), 136.8 (s, Ar), 151, 9 (s), 162.1 (ds, J _C F = 250HZ, 1 C, C4 ^{, m} ).

MS m / z (Cl, NH _3): 360 (28, M ^+), 269 (51), 237 (1 1), 167 (100), 139 (12), 91 (52), 65 (12).

HRMS (Cl, NH _3): MH ^+, Found .: 361, 1 171; calcd for C22H17FN2S: 362.1169.

-2-yl) thiazole-2 (3H) -ylidene) aniline 44

22 44

According to General Procedure B above, 30 mg of 22 was reacted with 2-iodothiophene for 24 hours. MPLC (LP: EtOAc 12: 1-0: 100 + 5% triethylamine) gave the title compound 44 as a brown oil (24 mg, 59% of theory).

R _f (PE / EtOAc 9: 1): 0.51

¹ H-NMR (200 MHz, CDCl ₃ ) δ: 5.09 (s, 2H, -CH ₂ ), 6.69 (s, 1 H, H 4), 6.82 (d, J = 3.7Hz, 1 H, H5 ^{, M} ), 6.92 (t, J = 3.6Hz, H4 '''), 7.10 (d, J = 7.3Hz, 4H, ArH), 7.32-7.39 ( m, 8H, ArH).

MS m / z (Cl, NH _3): 347 (43, M ^+), 257 (100), 225 (14), 213 (10), 167 (88), 127 (20), 91 (44).

HRMS (Cl, NH _3): MH ^+, Found .: 349.0832, Calcd for C20H16N2S2:. 349.0828.

Example 22 - Arylation of thiazolimine 33

 Preparation of N- (3-Benzyl-5- (2-fluoropyridin-3-yl) thiazol-2 (3H) -ylidene) aniline-4-carboxylic acid ethyl ester 46

33 46

33 46

 According to General Procedure B above, 60 mg of 33 was reacted with 2-fluoro-3-iodo-pyridine for 20 hours. MPLC gave the title compound 46 as a brownish oil (60 mg, 77% of theory).

activity tests

 Preparation of cell samples

 The cell material was produced at the Institute of Pharmacology of the Medical University of Vienna. C2C12 mouse cells were used as a model to determine the differentiation-promoting properties of the compounds prepared as described above. For this purpose, the C2C12 cells were incubated for two days in culture dishes with growth medium (Dulbecco's modified Eagle's medium, DMEM, with 4.5 g / l glucose, 4 mM L-glutamine, 50 U / ml penicillin, 50 pg / ml streptomycin and 20% fetal calf serum) were incubated before the test compounds were added in differentiation medium (substantially identical to the growth medium, except that it contained 2% horse serum instead of fetal calf serum).

For this purpose, the compounds were first dissolved in DMSO, and these solutions were added to aliquots of the differentiation medium. As a comparison was pure DMSO dissolved in differentiation medium. First, 5 μΜ solutions of the compounds were used, each experiment being carried out in duplicate.

After the two-day incubation of the cells in growth medium, the latter was removed, after which 3 ml of the above solutions of the respective compound and DMSO in differentiation medium were added to each cell culture. Subsequently, the cells were incubated at 37 ° C for 7 days. Meanwhile, the medium was exchanged twice, after 3 and 5 days. After 7 days, the cells were harvested, the medium was removed, and the cells were washed with PBS (phosphate buffered saline) to remove any residual differentiation medium. This is of essential importance as the medium contains proteins that could influence subsequent proteome analyzes.

After the washing step, 800 μΙ PBS was added to each cell culture and the cells were scraped from the plates and suspended in PBS. These suspensions were transferred to Eppendorf tubes and centrifuged. The PBS supernatant was removed and the cell pellets were frozen at -80 ° C. The washing and centrifuging steps were carried out at 4 ° C.

Observation of morphological changes

 Morphological changes of the cells were documented by photomicrographs of the cell cultures after 3, 5 and 7 days. Normally, C2C12 cells in the differentiation medium differentiate into mature skeletal muscle cells (long, multinucleated "myotubes"). Already after 3 days, some of the compounds sometimes appeared to increase the rate of differentiation considerably compared to controls containing only differentiation medium and DMSO.

Unfortunately, some of the test compounds also developed more or less toxicity to the cells, as evidenced by dead cells. showed material in the samples. Both effects, ie acceleration of differentiation and toxicity, were enhanced during the following days.

The results of the tests are listed in Table 1 below, both for differentiation acceleration and for toxicity "-" in each case for "no effect", "+" for "weak effect" and "++" for "strong effect" , Compounds which did not effect accelerated differentiation are designated as Comparative Examples "V", while all others are designated "Inventive" "E". Of these, those with as little or no toxicity as possible are naturally preferred.

Table 1

Verbindung DifferenzierungsToxizität beschleunigung

Connection differentiation toxicity acceleration

-V

 28 -E ++

 +

++ ++

31b-E ++ +

Verbindung DifferenzierungsToxizität -E ++ +

Compound differentiation toxicity

 acceleration

 39 - E ++ -

^Q >

41 - E ++ +

 42 - E + -

43 - E ++ -

46 - E ++ ++

As can be seen from Table 1, the ten thiazolimines prepared show consistently differentiation-accelerating effect, while of the ten Thiazolamines three compounds show no such effect. It is also clear from the table that both the nature of the aryl groups on the thiazole ring, their substituents, and the position of these substituents play an essential role - both in terms of efficacy and toxicity. More electron-attractive substituents, such as nitro or fluorine, also appear to exert a correspondingly stronger activating influence on the aromatic ring system. Corresponding tests with other new compounds as well as with other cell types are currently being conducted by the inventors.

Thus, the present invention provides a group of compounds capable of accelerating the differentiation of cells which are accessible in a relatively simple and inexpensive manner and which show little to no toxicity.

Claims

1. Use of a thiazolamine derivative for accelerating the differentiation of cells, characterized in that a 2-thiazolamine derivative of the general formula below is used:

wherein: R 1 to R _{3 are} independently selected from hydrogen, benzyl, aryl and heteroaryl or absent, with at most one of the radicals R 1 to R _{3 being} hydrogen and R _{4 is selected} from benzyl, aryl and heteroaryl, where Ri to R _{4 are} independently of one another optionally substituted by one or more substituents selected from halogen (F, Cl, Br, I), hydroxy, C 1-3 -alkyl, C 1-3 -alkoxy, carboxy, C 1-3 -alkylcarbonyl, Ci _3- alkoxycarbonyl, nitro, amino, mono- and di-Ci-3-alkylamino and cyano, in which the alkyl groups are optionally halogenated, are substituted;  Each represents a single bond or a double bond;  n = 0 to 2; where, in the case n = 1, the radical R4 can be bonded to position 4 or 5 of the thiazole ring and, in the case n = 2, the radicals R ₄ are bonded to the positions 4 and 5 of the thiazole ring. 2. Use according to claim 1, characterized in that the thiazole-amine derivative is selected from: N-benzyl-N, 5-diphenylthiazolamine (23), N-benzyl-N-phenyl-5- (2-fluoropyridin-3-yl) -thiazolamine (24),  Ethyl 4- (2- (N-benzyl-N-phenylamino) thiazol-5-yl) benzoate (25), N-benzyl-N-phenyl-5- (m-nitrophenyl) -thiazolamine (28), N-benzyl-N-phenyl-5- (p -fluorophenyl) -thiazolamine (31a), N-benzyl-N-phenyl-4,5-bis (p-fluorophenyl) thiazolamine (31b), N-benzyl-N-phenyl-5- (thiophen-2-yl) -thiazolamine (32), N- (3-benzyl-5-phenylthiazol-2 (3H) -ylidene) aniline (34), N- (3-Benzyl-5- (2-fluoropyridin-3-yl) thiazole-2 (3H) -ylidene) aniline (35), N- (3-Benzyl-5- (2-fluoropyridin-4-yl) thiazole-2 (3H) -ylidene) aniline (36), Ethyl 4- (3-benzyl-2- (phenylimino) -2,3-dihydrothiazol-5-yl) benzoate (37), N- (3-benzyl-5-o-tolylthiazol-2 (3H) -ylidene) aniline ( 38) N- (3-benzyl-5-p-tolylthiazol-2 (3H) -ylidene) aniline (39), N- (3-benzyl-5- (4-nitrophenyl) thiazole-2 (3H) -ylidene) aniline (41),  N- (3-Benzyl-5- (4-methoxyphenyl) thiazole-2 (3H) -ylidene) aniline (42), N- (3-Benzyl-5- (4-fluorophenyl) thiazole-2 (3H) -ylidene) aniline (43) and N- (3-Benzyl-5- (2-fluoropyridin-3-yl) thiazole-2 (3H) -ylidene) aniline-4-carboxylic acid ethyl ester (46). 3. Use according to claim 2, characterized in that the thiazole-amine derivative is selected from: N-benzyl-N, 5-diphenylthiazolamine (23), N-benzyl-N-phenyl-5- (m-nitrophenyl) -thiazolamine (28), N- (3-benzyl-5-phenylthiazol-2 (3H) -ylidene) aniline (34), N- (3-benzyl-5-p-tolylthiazol-2 (3H) -ylidene) aniline (39), N- (3-Benzyl-5- (4-methoxyphenyl) thiazole-2 (3H) -ylidene) aniline (42) and N- (3-Benzyl-5- (4-fluorophenyl) thiazole-2 (3H) -ylidene) aniline (43). 4. Use according to one of claims 1 to 3, characterized in that the thiazolamine derivative is selected from: N-benzyl-N-phenyl-5- (m-nitrophenyl) -thiazolamine (28), N- (3-Benzyl-5-p-tolylthiazol-2 (3H) -ylidene) aniline (39) and N- (3-Benzyl-5- (4-fluorophenyl) thiazole-2 (3H) -ylidene) aniline (43). 5. Use according to one of the preceding claims, characterized in that the differentiation of C2C12 cells is accelerated by the thiazolamine derivative.