CZ20001285A3 - Substituted 4-aminothiazol-2-yl compounds functioning as inhibitors of cyclin-dependent kinases - Google Patents

Abstract

The present invention relates to aminothiazole compounds of the formula wherein R 1 is substituted or unsubstituted C 1-6 -alkyl, C 1-6 -alkenyl; C 1-6 -alkynyl; C 1-6 -alkoxy; Cpe-alcohol; cycloalkyl; aryl; carbonyl; ether; (C 1-6 -alkyl) -carbonyl; (C 1-6 -alkyl) -aryl; (Ci.6- alkyl) -cycloalkyl; (C 1-6 -alkyl) - (C 1-6 -alkoxy); aryl- (C1-6- alkoxyl); thioether, thiol; and sulfonyl; and R 2 is carbocyclic or heterocyclic, monocyclic or fused or uncondensed polycyclic, ring structure having a substituent at a position adjacent to the point of attachment, wherein the ring structure optionally further substituted; or pharmaceutically acceptable salts of the compound of formula (I), or a prodrug or pharmaceutically active metabolite of the compound of formula (I) or a pharmaceutically acceptable salt thereof; inhibiting cyclin-dependent kinases (CDKs) such as CDK1, CDK2, CDK4 and CDK6. The invention also relates to a therapeutic or prophylactic use of the pharmaceutical compositions containing such compounds and methods of treating cancer tumors and other diseases by administering an effective amount of these compounds.

Description

Substituted 4-aminothiazol-2-yl compounds as inhibitors of cyclin-dependent kinases

Technical field:

The present invention relates to pharmaceutical compositions comprising aminothiazole compounds for inhibiting cyclin-dependent kinases (CDKs) such as CDK1, CDK2, CDK4, and CDK6. The invention is also directed to the therapeutic or prophylactic use of pharmaceutical compositions comprising these compounds, and to methods of treating cancer and other diseases by administering an effective amount of the compounds.

BACKGROUND OF THE INVENTION:

Uncontrolled cell proliferation is a sign of cancer. Cell proliferation in response to various stimulations is demonstrated by deregulation of the cell division cycle, the process by which cells multiply and divide. Tumor cells typically have damage to genes that directly or indirectly regulate cell cycle progression of division.

CDKs are a class of enzymes that play a crucial role in regulating transitions between different phases of the cell cycle, such as progression from G1 quiescent (gap between mitosis and onset of DNA replication for a new round of cell division) to S (active DNA synthesis period), or progression from G2 to M phase in which active mitosis and cell division occur. See, e.g., the article in Science, Vol. 274 (1996), pp. 1663-1677; and Ann.Rev.

Cell Dev. Biol., Vol. 13 (1997), pp. 261-291. CDK complexes are formed by linking the cyclin regulatory subunit (such as cyclin A,

B1, B2, D1, D2, D3 and E) and catalytic kinase subunits (such as cdc2 (CDK1), CDK2, CDK4, CDK5 and CDK6). As indicated by the title, e * shows an absolute dependence on the cyclin subunit to phosphorylate their target substrates, and different kinase / cyclin pair functions to regulate progression through specific parts of the cell cycle.

D cyclins are sensitive to extracellular growth signals and activate in response to mitogens during the G1 phase of the cell cycle. CDK4 / cyclin D plays an important role in the progression of the cell cycle by phosphorylation and thus inactivation of the retinoblastoma protein (Rb). Hypophosphorylated Rb binds to the family of transcriptional regulators, but during hyperphosphorylation of Rb by CDK4 / cyclin D, these transcription factors are released to activated genes whose products are responsive to S phase progression. Rb phosphorylation and inactivation of CDK4 / cyclin D allows the cell to pass through the D1 phase restriction point, whereupon the sensitivity to extracellular growth or inhibitory signals is lost and the cell is surrendered to cell division. During the next G1, Rb is also phosphorylated and inactivated by CDK2 / cyclin E, and recent evidence suggests that CDK2 / cyclin E can also regulate S phase progression in a parallel way that is independent of Rb phosphorylation (see Lukas et al., Cyclin E-induced With Phase Without Activation of the pRb / E2F Pathway, Genes and Dev., Vol. 11 (1997), pp. 1479-1492).

Progression from G1 to S phase, completed by CDK4 / cyclin D and CDK4 / cyclin E, is susceptible to a change in the growth regulatory mechanism, both negative and positive. Growth stimuli, such as mitogens, cause increased cyclin D1 synthesis and thus increased functional CDK4. In contrast, cell growth can be controlled in response to DNA damage or negative growth stimuli by inducing endogenous inhibitory proteins. These naturally occurring protein inhibitors include the p21 ^{WAF / CIP1} , the p27 ^KIPI , and the p16 ^INK4 family, the latter of which exclusively inhibit CDK4 (see Harper, Cyclin Dependent Kinase Inhibitors, Cancer Surv., Vol.

(1997), pp. 91-107). Deviations in this control system, especially those affecting CDK4 and CDK2 function, are involved in cell growth to a highly proliferative state indicative of cancer, such as hereditary melanomas, esophageal carcinomas, and pancreatic cancers (see, e.g. Peters, Genetic Alterations of Cyclins, Cyclin-Dependent Kinases, and CDK Inhibitors in Human Cancer, Adv. Cancer Res., Vol. 68 (1996), pp. 67-108, and Kamb et al., A Cell Cycle Regulator Potentially Involved in Genesis of Many Tumor Types, Science, Vol 264 (1994), pp 436-440). Cyclin D overexpression is associated with esophageal, breast, and squamous cell carcinomas (see, eg, DelSal et al., Cell Cycle and Cancer: Critical Events at the G1 Restriction Point, Critical Rev. Oncogenesis, Vol. 71 (1996), p. 127-142). Genes encoding CDK4-specific inhibitors of the p16 family often have inherited deletions and mutations of melanoma, gliomas, leukemias, sarcomas and pancreatic cancers, non-small lung cell cancers, and cancers of the clavicle and neck (see Nobori et al., Deletions of the Cyclin Dependent Kinase-4 Inhibitor Gene in Multiple Human Cancers, Nature, Vol 368 (1994), p.

753-756). Amplification and / or overexpression of cyclin E was also observed in a wide variety of solid tumors, and elevated cyclin E levels were related to poor prognosis. Furthermore, cellular levels of CDK inhibitor p27, which acts as both a substrate and a CDK2 / cyclin E inhibitor, are abnormally low in breast, colon and prostate tumors, and p27 expression levels are inversely related to the disease state (see Loda et al., Increased Proteasome- Dependent Degradation of the Cyclin-Dependent Kinase Inhibitor p27 in Aggressive Colorectal Carcinomas, Nature Medicine, Vol 3 (1997), pp. 231-234). The p21 proteins also appear to transmit the p53 tumor suppressive signal to CDK; thus, a p53 mutation in approximately 50% of human cancers may indirectly result in deregulation of CDK activity.

The emerging data provide strong support for the use of compounds inhibiting CDK and CDK4 and CDK2, in particular, as anti-proliferative agents. Certain biomolecules were for these ρ ρ ρ · * Ρ Γ ρ ρρρρ ρ Ρ Ρ · ρ r ρ rrpp ρ rpppp Ρ '· rrr ^r r Γ

- C P prP <- r ~ r

P - Ρ ρ Ρ Ρ Γ - r · r ρ Γ '

purposes designed. For example, U.S. Patent No. 5,621,082 to Xiong et al. Discloses a nucleic acid encoding a CDK6 inhibitor, and European Patent Application EP 0 666 270-A2 discloses peptides and mimetic peptides that act as inhibitors of CDK1 and CDK2. More small molecules have been identified as CDK inhibitors (see, recently, Webster, The Therapeutic Potential of Targeting the Cell Cycle, Exp. Opin. Invest. Drugs, Vol. 7 (1998), pp. 865-887). Flavon flavopiridol shows slight selectivity for inhibition of CDK over other kinases, but inhibits

CDK4, CDK2 and CDK1 equivalently, with IC _{50 s} in the range 0.1-0.3 μΜ. Flavopiridol is currently in Phase II clinical trials as an oncological chemotherapeutic agent (Sedlaček et al.,

Flavopiridol (L86-8275; NSC 649890), A New Kinase Inhibitor for Tumor Therapy, Int. J. Oncol., Vol. 9 (1996), pp. 1143-1168). Flavopiridol analogs are the subject of other publications, e.g., U.S. Patent No. 5,733,920 to Mansuri et al., (International Application No. WO 97/16447) and International Applications WO 97/42949 and WO 98/17662. Results with purine-based derivatives are described by Schow et al., Bioorg. Copper. Chem. Lett., Vol. 7 (1997), pp. 2697-2702; Grant et al. Why. Amer. Assoc. Cancer Res. 39 (1998) Abst. 1207; Legravend et al., Bioorg. Copper. Chem.

Lett., Vol. 8 (1998), pp. 793-798; Gray et al., Science, Vol. 281 (1998), pp 533-538; and Furet et al., 216th ACS Natl. Mtg. (Aug. 23-27, 1998, Boston), Abst. MEDI-218 Further, the following publications disclose some pyrimidines that inhibit cyclin-dependent kinases and growth factor-mediated kinases: International Application WO 98/33798; Ruetz et al., Proc. Amer. Assoc. Cancer Res. 39 (1998) Abst. 3796; and Meyer et al., Proc. Amer. Assoc. Cancer Res. 39 (1998) Abst. 3794.

Thus, there is a need for small molecule compounds that can be readily synthesized and are potential inhibitors for one or more CDK or CDK / cyclin complexes. Since CDK4 can serve as a general activator of cell division in most cells, and because the CDK4 / cyclin D and CDK2 / cyclin E complexes determine the early G1 phase of the cell cycle, there is a need to obtain effective and specific inhibitors of CDK4 and / or CDK2 for treating one or more tumor types. .

SUMMARY OF THE INVENTION

One object of the invention is to provide compounds and drug compositions that inhibit the activity of one or more CDKs, such as CDK2, CDK4 and / or CDK6, or cyclin complexes thereof. Another object is to provide an effective method of treating cancer symptoms by CDK inhibition, preferably by inhibiting CDK4 or CDK4 / D-type cyclin complexes and / or CDK2 or CDK2 / E-type cyclin complexes. Another object is to provide pharmaceutical compositions comprising compounds effective to block the transition of cancer cells to their proliferative phase. These and other objects and advantages of the invention, which will be explained in the following detailed description, are obtained by the cell cycle control / regulating agents of the invention described below.

In general terms, the invention relates to pharmaceutical compositions comprising:

(a) a cell cycle regulator selected from (i) a compound of formula I nh ₂ , O (I)

R ¹ is a substituted or unsubstituted group selected from: (C 1-8 -alkyl, such as methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl or tert-butyl);

C1-6-alkenyl; C 1-8 -alkynyl; C 1-6 -alkoxy; C 1-4 -alcohol; carbocyclic or heterocyclic cycloalkyl, which may be monocyclic or fused or non-fused polycyclic, such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl; or heterocycloalkyl, which may be monocyclic or fused or non-fused polycyclic, such as pyrrolidinyl, piperidinyl, morpholinyl; carbocyclic or heterocyclic, monocyclic or fused or non-fused polycyclic aryl such as phenyl, naphthyl, pyrrolyl, indonyl, furanyl, thiophenyl, imidazolyl, oxazolyl, isoxazolyl, thiazolyl, triazolyl, tetrazolyl, pyrazolyl, pyridinyl, quinolinyl, isoquinolinyl, isoquinolinyl, isoquinolinyl, isoquinolinyl, isoquinolinyl, pyridazinyl, pyrimidinyl, benzimidazolyl, benzothiophenyl or benzofuranyl; carbonyl such as carboxyl, ester, aldehyde or ketone; ether;

(C 1-8 -alkyl) -carbonyl; (C 1-6 -alkyl) -aryl; (Cx.g-alkyli cycloalkyl; (C ^ g-alkyl) - (C _L .g-alkoxy); aryl- (C ^ .g-alkoxy); thioethers such aryl-aryl)., Cycloalkyl- S-aryl, cycloalkyl-S-cycloalkyl or dialkyl sulfide; thiol; and sulfonyl; and

R ² is substituted or unsubstituted: carbocyclic or heterocyclic, monocyclic or fused or non-fused polycyclic, ring structure;

wherein each possible R ¹ and R ² is independently halogen such as chlorine, bromine, iodine or fluorine; oxygen (= O); haloalkyl such as trifluoromethyl; C 1-6 -alkyl; C 1-8 -alkenyl; C 1-8 -alkynyl; hydroxyl; C 1-6 -alkoxy; carbocyclic cycloalkyl, which may be monocyclic or fused or non-fused polycyclic, such as cyclopropyl, cyclobutyl, reerccerrr (rcf <- r - <cyclic pentyl, or cyclohexyl; or heterocycloalkyl, which may be monocyclic or fused or unfused polycyclic, such as pyrrolidinyl , piperidinyl, piperazinyl, morpholinyl or thiazinyl;

heterocyclic, monocyclic or fused or non-fused polycyclic aryl, such as phenyl, naphthyl, pyrrolyl, indolyl, furanyl, thiophenyl, imidazolyl, oxazolyl, isoxazolyl, thiazolyl, triazolyl, tetrazolyl, pyrazolyl, pyridinyl, quinolinyl, pyridinazinyl, acridinyl, acridinyl, acridinyl, acridinyl , benzimidazolyl, benzothiophenyl or benzofuranyl; amino - primary, secondary or tertiary; nitro; thiol; thioether; imine; cyano; amido; phosphonato; phosphine; carboxyl; thiocarbonyl; sulfonyl; sulfonamide; ketone; aldehyde; or an ester;

(ii) pharmaceutically acceptable salts of the compounds of Formula I; and (iii) prodrugs and pharmaceutically active metabolites of the compounds of Formula I, or pharmaceutically acceptable salts thereof; and (b) a pharmaceutically acceptable carrier.

In another general aspect, the invention relates to pharmaceutical compositions comprising:

(a) a cell cycle regulator selected from (i) a compound of Formula I:

r Γ Γ r r c c p p p p A p p p p p p p p p r r r r where:

R ¹ is selected from

R ² is substituted or unsubstituted: carbocyclic or heterocyclic, monocyclic or fused or non-fused polycyclic, ring structure;

wherein each possible R ² is independently halogen such as chlorine, bromine, iodine or fluorine; oxygen (= O); haloalkyl such as trifluoromethyl; C _1-6 -alkyl; _{C> 6} -alkenyl; C 1-8 -alkynyl; hydroxyl; C 1-8 -alkoxy; carbocyclic cycloalkyl, which may be monocyclic or fused or non-fused polycyclic, such as cyclopropyl, cyclobutyl, cyclopentyl, or cyclohexyl; or heterocycloalkyl, which may be monocyclic or fused or non-fused polycyclic, such as pyrrolidinyl, piperidinyl, piperazinyl, morpholinyl or thiazinyl; carbocyclic or heterocyclic, monocyclic or fused or non-fused polycyclic aryl, such as phenyl, naphthyl, pyrrolyl, indolyl, furanyl, thiophenyl, imidazolyl, oxazolyl, isoxazolyl, thiazolyl, triazolyl, tetrazolyl, pyrazolyl, pyridinyl, quinolinyl, pyridinyl, pyridininyl, pyridinyl, pyridininyl, pyridinyl, pyridininyl , pyrimidinyl, benzimidazolyl, benzothiophenyl or benzofuranyl; amino - primary, secondary or r t - erf tertiary; nitro; thiol; thioether; imine; cyano; amido; phosphonato; phosphine; carboxyl; thiocarbonyl; sulfonyl; sulfonamide; ketone; aldehyde; or an ester;

(ii) pharmaceutically acceptable salts of the compounds of Formula I; and (iii) prodrugs and pharmaceutically active metabolites of the compounds of Formula I, or pharmaceutically acceptable salts thereof; and (b) a pharmaceutically acceptable carrier.

These compositions are useful as inhibitors of mammalian CDK / cyclin complexes, insect CDKs or fungal CDK complexes. These compositions are also suitable for regulating proliferation, differentiation and / or apoptosis. Thus, one general aspect of the invention is directed to pharmaceutical compositions comprising pharmaceutically effective amounts of cell cycle regulating agents.

In a preferred embodiment, the invention is directed to a potent, cell-cycle regulating agent wherein R ² in Formula I is an ortho-substituted aryl ring structure (such as o-substituted phenyl). Particularly preferred among these compounds are those in which R ² is ^an o-disubstituted phenyl.

The invention further relates to methods of using cell cycle regulating agents for treating diseases or disorders mediated by CDK inhibition, in particular those mediated by CDK4 and / or CDK2 inhibition. More preferably, the invention is directed to methods of treating malignant or cancer-like diseases by administering a pharmaceutical composition comprising a cell cycle regulating agent. Further, the invention relates to the use of cell cycle regulating agents to prevent or treat fungal infections.

Other aspects, advantages and suitable features of the invention will be apparent from the detailed description below.

Detailed description and preferred embodiments of the invention

In general terms, the invention relates to pharmaceutical compositions comprising:

(a) an amount of a cell cycle regulator effective to inhibit CDK, wherein the cell cycle regulator is:

(i) a compound of formula I:

NH ₂ (I) wherein R ¹ is a substituted or unsubstituted group selected from: C 1-4 -alkyl; C ₁ . _6- alkenyl; C ₁ . _6- alkynyl; C 1-8 -alkoxy; carbocyclic or heterocyclic, monocyclic or fused or non-fused polycyclic, cycloalkyl; carbocyclic or heterocyclic, monocyclic or fused or non-fused polycyclic, aryl; carbonyl; ether; (C _1-6 -alkyl) -carbonyl; (C 1-8 -alkyl) -aryl; (C 1-8 -alkyl) cycloalkyl; (C 1-6 -alkyl) - (C 1-8 -alkoxy); aryl- (C 1-4 -alkoxy) for thioether and sulfonyl;

and

R ² is a substituted or unsubstituted, carbocyclic or heterocyclic, monocyclic or fused or non-structure;

wherein each possible R ¹ and R ² is independently halogen; haloalkyl; C ₁ . _6- alkyl; C ₁ . ₆ -alkenyl; C,. _6- alkynyl;

r r

Γ Γ '

hydroxyl; C 1-8 -alkoxy; amino; nitro; thiol; thioether; imine; cyano; amido; phosphonato; phosphine; carboxyl; thiocarbonyl; sulfonyl; sulfonamide; ketone; aldehyde; or an ester;

(ii) a pharmaceutically acceptable salt of a compound of Formula I; or (iii) a prodrug and a pharmaceutically active metabolite of a compound of Formula I, or a pharmaceutically acceptable salt thereof; and (b) a pharmaceutically acceptable carrier.

In another general embodiment, each optional substituent for R ¹ and R ² may be independently selected from, together with the groups identified above, the following groups: oxygen, carbocyclic or heterocyclic, monocyclic or fused or non-fused polycyclic, cycloalkyl; and carbocyclic or heterocyclic, monocyclic or fused or non-fused polycyclic, aryl. These substituents may optionally be further substituted by substituents selected from these groups.

Examples of the R ¹ moiety include substituted or unsubstituted aryl and alkyl such as phenyl, pyridyl, benzimidazole, benzyl and C 1-6 -alkyl. Preferably, these groups have one or more substituents selected from: halogen; oxygen; haloalkyl; C 1-6 -alkyl; cycloalkyl; heterocycloalkyl; aryl; hydroxyl; C 1-8 alkoxy; amino; nitro; thioether; cyano; amido; carboxyl; sulfonamido; ketone; aldehyde; and ester.

Other preferred moieties for R ¹ are phenyl groups substituted with an alkylamine or pyridine group having optional substituents selected from the group described above for R ¹ . The alkylamino substituent may be a 5-7-membered heterocycloalkyl optionally containing, together with the nitrogen atom in the ring, one or more heteroatoms selected from N, O and S.

Examples of such preferred R ¹ groups include phenyl substituted in the para-position with heterocycloalkyl, for example piperidinyl, piperazinyl, thiazinyl or morpholinyl, or. pyridyl.

The following are examples of preferred R ¹ groups:

e · c c r r r13

Other particularly preferred R ¹ groups include phenyl groups substituted with a carbonyl or sulfonamide moiety, the carbonyl carbon and the sulfonamide nitrogen are preferably further substituted. The following are examples of preferred R ¹ groups:

wherein R ³ is selected from C 1 -C ₆ alkyl, C 1 -C ₆ alkoxy, aryl, aryloxy and an amine.

Other preferred examples for the R ¹ moiety include substituted or unsubstituted phenyl, alkylbenzyl, alkyl, benzylcarboxylate, benzyloxyphenyl, dimethylaminophenyl, pyridinyl, phenethyl, alkylcarboxyl, alkylpiperidinyl, phenylamino, cyclohexyl, benzylcarboxyalkyl, benzylnitro, phenyl-alkoxy, ethylbenzoate, benzylcarboxylate, benzylcarboxy, benzylcarboxy, benzylcarboxy, , benzyldimethylamino, pyridinylsulfanyl, cyanobenzyl, and phenylsulfamyl.

In preferred embodiments, R ² in Formula I is a bulky group such as a substituted or unsubstituted carbocyclic or heterocyclic monocycle, or a substituted or unsubstituted fused or non-fused carbocyclic or heterocyclic polycycle. Even more preferably, R ^{2 is} substituted (carbo or poles) - (monocycle or polycycle); even more preferably, R ^{2 is} such a cyclic ring structure carrying a substituent at a site adjacent or adjacent to the attachment site (at the core structure).

r ι14

For example, preferred species for R ² include an ortho-substituted aromatic ring structure such as o-substituted phenyl or thienyl, or 1,2-substituted cycloalkyl or cycloalkenyl ring structure such as 2-substituted cyclopent-enyl. Particularly preferred examples for the R ² moiety include substituted or unsubstituted: o-halophenyl (e.g.

O-fluorophenyl, o-chlorophenyl, o-iodophenyl, or o-bromophenyl), o-nitrophenyl, o-aminophenyl, oC-alkylphenyl, o-C 1-8 -alkoxyphenyl (e.g. o-methoxyphenyl or o-ethoxyphenyl), o-C 1-8 -alkoxybenzothiophenyl, o-methylthiophenyl, benzonitrile and carboxybenzyl. Particularly preferred examples for the R ² moiety also include ortho-disubstituted aryl, for example 2,6-dihalophenyl (e.g.

2,6-difluorophenyl) and 2-halo-6-trifluoromethylphenyl (e.g., 2-fluoro-6-trifluoromethylphenyl). Compounds of formula I wherein R ² is a 1,2-substituted cyclic ring structure, preferably having one or more additional substituents, such as an ortho-substituted aryl having an additional substituent at the para-position, have surprisingly been found as potent CDK inhibitors.

Particularly preferred examples of compounds of formula I include:

nh ₂ sec @ -1

O, a

LA ch ₃ e 'ri * erf' eeer

Another preferred compound of formula I is:

Other particularly preferred examples of compounds of Formula I include:

h ₂ n - | O

h ₃ c τ & λ

•WITH.

The H ₃ CF pharmaceutical compositions of the invention may, alternatively or together with a compound of Formula I, contain as an active ingredient a pharmaceutically acceptable salt of a compound of Formula I, or a prodrug or pharmaceutically active metabolite of such a compound or salt. These compounds, salts, prodrugs and metabolites are sometimes collectively referred to herein as cell cycle regulators.

The compositions according to the present invention inhibit the kinase activity of CDK / cyclin complexes, such as that which is active in _the G or G i stage of the cell cycle, such as CDK2, CDK4 and / or CDK6 complexes. Preferred compositions of the invention comprise a cell activity regulator having an inhibitory constant against CDK4 or a CDK4 / D-type cyclin complex of about 1 μΜ or less, more preferably about 500 nM or less, and even more preferably about 200 nM or less, and most preferably about 100 nM or less. Particularly preferred compounds of the invention include those having a CDK4 / cyclin D3 ( _? CDK4 / D3) inhibition constant of about 100 nM or less. Other preferred compositions of the invention comprise cell cycle regulators having a CDK2 inhibitory constant or CDK2 / E-type cyclin complex of about 1 μΜ or less, more preferably about 500 nM or less, even more preferably about 200 nM or less, and most preferably about 100 nM or less. nM or less.

Certain compounds of formula I may exist in various stereoisomeric or tautomeric forms. The present invention encompasses all such CDK-inhibiting compounds, including the active compounds in the form of substantially pure enantiomers, racemic mixtures and tautomers.

The term pharmaceutically acceptable refers to a pharmacologically acceptable and substantially non-toxic cell cycle regulator for the subject to be administered.

Pharmaceutically acceptable salts include conventional acid addition salts or base addition salts formed from suitable non-toxic organic or inorganic acids or inorganic bases. Exemplary acid addition salts include those derived from inorganic acids such as hydrochloric acid, hydrobromic acid, hydroiodic acid, sulfuric acid, sulfamic acid, phosphoric acid, and nitric acid, and derived from organic acids such as p-toluenesulfonic acid, methanesulfonic acid, ethanedisulfonic acid, isethionic acid, oxalic acid, p-bromophenylsulfonic acid, carbonic acid, succinic acid, citric acid, benzoic acid, 2-acetoxybenzoic acid, acetic acid, phenylacetic acid, propionic acid, glycolic acid, stearic acid, lactic acid, malic acid, tartaric acid, rrerrr ascorbic acid, maleic acid, hydroxymaleic acid, glutamic acid, salicylic acid, sulfanilic acid and fumaric acid. Examples of basic addition salts include salts derived from ammonium hydroxides (i.e., quaternary ammonium hydroxide as well as tetramethylammonium hydroxide), salts derived from inorganic bases such as alkali or alkaline earth metal hydroxides (such as sodium, potassium, lithium, calcium or magnesium salts), and salts derived from organic bases such as carbonates, bicarbonates, amines, benzylamines, piperidines and pyrrolidines.

The term prodrug refers to a metabolic precursor of a compound of Formula I (or a salt thereof) that is pharmaceutically acceptable. A prodrug may be inactive when administered to a subject, but converted in vivo to the active compound of Formula I. The term active (active) metabolite refers to a metabolic product of a compound of Formula I that is pharmaceutically acceptable and active. Prodrugs and active metabolites of compounds of Formula I may be determined by techniques known in the art.

The cell cycle regulating agents of the present invention are useful as pharmaceutical agents for the treatment of proliferative disorders in mammals, particularly humans, noticeable inadvertent proliferation of endogenous tissue. The compounds of formula I can be used to treat subjects having diseases associated with excess cell proliferation, i.e., cancers, psoriasis, immunological disorders, including unwanted leukocyte proliferation, and restenosis and other smooth muscle diseases. Furthermore, these compounds can be used to prevent de-differentiation of post-mitotic tissue and / or cells.

The pharmaceutical compositions or compositions of the invention include a pharmaceutically acceptable carrier and an effective amount of at least one cell cycle regulating agent. The term "effective amount" refers to an amount that significantly inhibits proliferation and / or prevents de-differentiation of a eukaryotic cell, i.e., a mammalian, insect, plant or fungal cell, and is effective at the indicated use, i.e., for a particular therapeutic treatment.

Of course, the particular dosage amount of the cell cycle regulator to be administered to obtain therapeutic or inhibitory effects may be determined by a method known in the art according to the particular circumstances of the case, including, for example, the particular agent to be administered, the route of administration, the condition to be administered. and the subject or host to be treated. For example, the total daily dose of the cell cycle regulator to be administered in single or multiple doses comprises a dose amount of from about 0.01 mg / kg body weight to about 50 mg / kg body weight.

The cell cycle regulator of the invention may be administered by any suitable route, i.e. orally, rectally, transdermally, subcutaneously, intravenously, intramuscularly, or intranasally. Cell cycle regulating agents are preferably formulated prior to administration into compositions suitable for the desired route of administration.

The pharmaceutical composition or composition of the invention comprises an effective amount of a cell cycle regulating agent and a pharmaceutically acceptable carrier such as a diluent or excipient for the agent. When the carrier serves as a diluent, it may be a solid, semi-solid, or liquid material acting as a vehicle, excipient or environment for the active ingredient (s). The compositions of the invention may be made by mixing the active ingredient (s) with the carrier, or diluting it with the carrier, or enclosing or encapsulating the carrier so that they may be in the form of a capsule, sachet, paper container or the like. Exemplary ingredients, together with one or more cell activity regulating agents and other active ingredients, include Avicel (microcrystalline cellulose), starch, lactose, calcium sulfate dihydrate, gypsum, sucrose, talc, gelatin, agar, pectin, acacia, magnesium stearate, peanut oil, olive oil, glyceryl monostearate, Tween 80 (polysorbate 80), 1,3-butanediol, cocoa butter, beeswax, polyethylene glycol, sorbitan monostearate, polysorbate 60, 2-octyldodecanol, benzyl alcohol, glycine, sorbic acid, potassium sorbate, hydrogen phosphate sodium, sodium chloride, and water.

The compositions of the invention may be prepared in any possible form suitable for the desired type of administration. For example, the pharmaceutical compositions may be prepared in the form of tablets, pills, powders, lozenges, sachets, capsules, elixirs, suspensions, emulsions, solutions, syrups, aerosols (as solid or liquid), ointments (eg containing up to 10% by weight) cell cycle regulators), soft gel and hard gel capsules, suppositories, sterile injectable solutions, sterile packaged powders and the like.

The pharmaceutical composition of the invention comprises a cell cycle regulator and, preferably, one or more other active ingredients, such as a known antiproliferative agent, which is compatible with the cell cycle regulator and is suitable for the indication being treated. In a preferred embodiment, the pharmaceutical composition of the invention comprises an effective amount of a cell cycle regulator of Formula I as an active ingredient.

The compounds of the invention may be prepared by procedures analogous to those described in the following and wherein the examples are indicated by letters (i.e. A, B, C, D, E, F, G, H, J, K, L, M and N). ), defining schemes for general syntheses.

f. _rr r r · Γ RRR (. <- (*** r (r 0 r _f r r. fr rrr r <r <rr

* Example A (1):

(4-Amino-2-phenylaminothiazol-5-yl) - (3-nitrophenyl) methanone

Following the procedure of Gewald et al., J. Pract. Chem., Vol. 35 (1967), pages 97-104, sodium (188 mg, 8.20 mmol) was carefully dissolved in methanol (9 mL) at 0 ° C and allowed to warm to room temperature. The resulting solution was added portion-wise to a mixture of cyanamide (345 mg, 8.20 mmol) and phenyl isothiocyanate (0.98 mL, 8.2 mmol) while releasing heat. 2-Bromo-3'-nitroacetophenone (2.00 g, 8.2 mmol) was added, and the resulting mixture was stirred overnight at ambient temperature. The mixture was diluted with water (150 mL). The yellow-brown solid was filtered, washed with water and a small amount of ether, dried under vacuum and recrystallized from ethanol to give 2.17 g (52% yield) of the title compound as dark brown crystals, mp (mp, mp) Mp 186-187 ° C.

1 H NMR (DMSO-d ₆ ): _δ 10.95 (1H, t, J = 1.9 Hz), 8.54-8.22 (2H, bs), 8.34 (1H, ddd, J = 8) 2.2, 1.9, 0.9 Hz), 8.12 (1H, ddd, J =

8.2, 1.9, 0.9 Hz). 7.78 (H, t, J = 8.2 Hz), 7.62 (2H, d, J = 7.8 Hz), 7.36 (H, t, J = 7.8 Hz), 09 (1H, t, J = 7.8Hz).

ESIMS (MH ^&lt; ^{+ &gt;} ): 341.

Anal. calculated for C ₁₆ H ₁₂ N ₄ O ₃ S. EtOH: C, 55.94; H, 4.70; N, 14.50; S, 8.30. Found: C, 55.96; H, 4.73; N, 14.40; S, 8.29.

* Example A (2):

(4-Amino-2-phenylamino-thiazol-5-yl) - (4-nitrophenyl) methanone

Rc ρ ρ r rc rc rc rc rc rc rc rc rc rc rc rc í rc r r r r r

The title compound was prepared in a manner similar to that described in Example A (1). Phenylisothiocyanate and 2-bromo-4'-nitroacetophenone gave, after recrystallization from ethanol, 3.06 g (55% yield) of red-brown crystals, m.p. Mp 162-164 ° C.

¹ H NMR (DMSO-d ₆ ): δ 10.91 (1H, s), 8.23 (2H, bs), 8.30 (2H, d, J = 8.7 Hz), 7.90 (2H δ d, J = 8.7 Hz), 7.59 (2H, d, J = 7.5 Hz), 7.36 (2H, t, J = 7.5 Hz), 7.10 (1H, d , J = 7.5 Hz).

FABMS (MH +): 341.

Anal. calcd for C ₁₆ H ₁₂ N 'O ₃ S: C, 56.46; H, 3.55; N, 16.46; WITH,

9.42. Found: C, 56.54; H, 3.54; N, 16.52; S, 9.42.

* Example A (3):

(4-Amino-2- (pyridin-3-ylamino) thiazol-5-yl) -phenyl-methanone

The title compound was prepared in a manner similar to that described in Example A (1). Pyridin-3-yl-isothiocyanate and phenacyl chloride gave, after recrystallization from ethanol, 4.1 g (75% yield) of yellow crystals, m.p. 227-229 [deg.] C.

^{Σ Η} NMR (DMSO-d ₆₎ d 10.95 (1H, s), 8.82 (1H, d, J = 2.5), 8.28 (1H, dd, J = 4.7, 1 2 Hz), 8.23 (2H, bs), 8.12 (1H, ddd, J = 8.4, 2.8, 1.6 Hz), 7.68 (1H, d, J = 6, 9 Hz), 7.66 (1H, d, J = 7.8 Hz), 7.54-7.44 (3H, m), 7.39 (1H, dd, J = 8.4, 4.7 Hz).

HRFABMS: Calcd for C ₁₅ H ₁₃ N ₄ OS (MH ^+): 297.0810. Found:

ρ ρ ρ rrr Pi C <7 ^r * “ _r ι r Γ / - r pf rrc r Γ rrr ^r 'r ^{Γ r} r'' ^r

297.0515.

Anal. calculated for C ₁₅ H ₁₂ N ₄ OS. EtOH: C, 59.63; H, 5.30; N, 16.36; S, 9.36. Found: C, 59.62; H, 5.32; N, 16.43; S, 9.41.

* Example A (4):

(4-Amino-2-phenylamino-thiazol-5-yl) -pyridin-2-yl-methanone

The title compound was prepared in a manner similar to that described in Example A (1). Phenylisothiocyanate and 2- (2-bromoacetyl) pyridine (see Menassé et al., Helv. Chim. Acta, vol. 38 (1955), pp. 1289-1291; Imuta et al., J. Org. Chem., Vol. 45 (1980), pp. 3352-3355) gave, after recrystallization from 95% ethanol, 510 mg (71% yield) of brown needles, m.p. 181.5 183.0 ° C.

¹ H NMR (DMSO-d 6): δ 10.75 (1H, s), 8.92 (1H, bs), 8.66 (1H, ddd,

J = 5.1, 1.8, 1.2 Hz), 8.22 (1H, bs), 8.13 (1H, dt, J = 7.5, 1.2)

Hz), 8.01 (1H, dt, J = 7.5, 1.8 Hz), 7.69 (2H, dd, J = 7.5) 1,2 Hz), 7.54 (1H, ddd, J = 7.5, 5.1, 1.2 Hz), 7.36 (2H, t, J = 1 ') 7.5 Hz), 7.07 (1H, dt, J = 7.5, 1.2 Hz).

HRFABMS: Calcd. For C ₁₅ H ₁₃ N ₄ OS (MH ⁺ ): 297.0810. Found: 297.0818.

Anal. calculated for C ₁₅ H ₁₂ N ₄ O ₃ S. H ₂ O: C, 57.31; H, 4.49; N, 17.82; S, 10.02. Found: C, 57.31; H, 4.46; N, 17.80; S, 10.14.

* Example A (5):

Ethyl 4- [4-Amino-5- (2-nitro-benzoyl) -thiazol-2-ylamino] -benzoate

H ₃ CCH ^ 0 '

Γ <"

Γ C

The title compound was prepared in a manner similar to that described in Example A (1). 4-Carboethoxyphenylisothiocyanate and 2-bromo-2'-nitroacetophenone gave, after recrystallization from ethanol, 1.2 g (59% yield) of a yellow crystalline powder, m.p. Mp 262-265 ° C.

¹ H NMR (DMSO-d ₆ ): _δ 11.08 (1H, s), 8.12 (2H, bs), 8.08 (1H, d, J = 8.7 Hz), 7.93 (2H , d, J = 8.7 Hz), 7.82 (1H, dt, J = 7.2, 1.2 Hz), 7.77-7.66 (3H, m), 7.73 (1H, d, J = 8.7 Hz), 4.28 (2H, q, J = 7.2 Hz), 1.30 (3H, t, J = 7.2 Hz).

ESIMS (MH ^&lt; ^{+ &gt;} ): 413.

Anal. calculated for Ο ₁₉ Η _1Θ Ν ₄ Ο ₃ 3: C, 55.33; H, 3.91; N, 13.58; S, 7.77. Found: C, 55.22; H, 3.86; N, 13.48; S, 7.67.

* Example A (6):

(4-Amino-2- (2-methyl-1H-benzimidazol-6-ylamino) -thiazol-5-yl) - (2-nitrophenyl) -methanone

The title compound was prepared in a manner similar to that described in Example A (1).

6-Isothiocyanato-2-methyl-1H-benzoimidazole (see Galley et al., German Patent Application DE 2259220 (1973); CA No. 478781 (1983)) and 2-bromo-2'-nitroacetophenone gave, after recrystallization from ethanol 1.2 g (62% yield) of brown crystals, m.p. 190.0 192.5 ° C.

NMR (DMSO-d ₅ ): _δ 12.20 (1H, bs), 10.76 (1H, s), 8.10-8.76 (3H, m), 7.60 (1H, t, J) = 7.0 Hz), 7.70-7.58 (3H, m), 7.40 (1H, d, J = 8.4 Hz), 7.13 (1H, dd, J = 8.4, 1.6 Hz), 2.44 (3H, s).

FABMS (MH ^&lt; ^{+ &gt;} ): 395.

e e r r

Anal. calculated for C ₁₈ H ₁₄ N ₆ O ₃ S. H ₂ O: C, 52.42; H, 3.91; N, 20.38; S, 7.77. Found: C, 52.29; H, 3.89; N, 20.31; S, 7.86.

* Example A (7):

(4-Amino-2- (4-iodo-phenylamino) -thiazol-5-yl) - (2-nitrophenyl) methanone

The title compound was prepared in a manner similar to that described in Example A (1). 4-Iodophenyl isothiocyanate and 2-bromo-2'-nitroacetophenone gave 7.9 g (88% yield) of an orange-red powder, m.p. Mp 182-184 ° C.

¹ H NMR (DMSO-d 6): δ 10.89 (1H, s), 8.20 (1H, s), 8.50 (1H, d, J = 8.7 Hz), 7.80 (1H, dd) J = 8.7, 6.2 Hz), 7.72-7.62 (4H, m), 7.41 (2H, d, J = 8.7 Hz).

FABMS (MH ^&lt; ^{+ &gt;} ): 467.

Anal. calcd for C ₁₆ H _N N ₄ O ₃ SI: C, 41.21; H, 2.38; N, 12.02; S, 6.88; I, 27.22. Found: C, 41.34; H, 2.46; N, 12.07; S, 7.02;

I, 27.35.

* Example A (8):

(4-Amino-2- (4-nitro-phenylamino) -thiazol-5-yl) -phenyl-methanone

The title compound was prepared in a manner similar to that described in Example A (1). 4-Nitrophenylisothiocyanate and phenacyl chloride gave 2.5 g (60% yield) of solid,

R Γ r r f c c r r r - - -

* Γ Γ <* ^r fr 'yy-y-yy <. rrr <- rrrr i- cr · · r '·

m.p. 280.0-281.5 ° C.

1 H NMR (DMSO-d ₆ ): _δ 11.38 (1H, s), 8.30-8.18 (2H, bs), 8.23 (2H, d, J = 9.3 Hz), 7 87 (2H, d, J = 9.3 Hz), 7.69 (2H, dd, J = 7.8, 1.6 Hz), 7.56-7.44 (3H, m).

FABMS (MH ^&lt; ^{+ &gt;} ): 341.

Anal. Calcd. for C ₁₅ H ₁₂ N ₄ O ₃ S: C, 56.46; H, 3.55; N, 16.46; WITH,

9.42. Found: C, 56.40; H, 3.49; N, 16.40; S, 9.41.

* Example A (9):

(4-Amino-2- (1H-benzoimidazol-6-yl-amino) -thiazol-5-yl) - (2-nitrophenyl) -methanone

The title compound was prepared in a manner similar to that described in Example A (1). 6-Isothiocyanate-1H-benzoimidazole (see Boev et al., Pharm. Chem. J. (Eng. Trans.), Vol. 24 (1990), pp. 812-822) and 2-bromo-2'-nitroacetophenone gave After recrystallization from ethanol / methanol, 1.5 g (83% yield) of a red-brown amorphous powder, mp 249-255 ° C.

¹ H NMR (DMSO-d 6): δ 12.50 (1H, bs), 10.84 (1H, s), 8.20 (1H, s),

8.60 (2H, bs), 8.03 (1H, d, J = 8.1 Hz), 7.88-7.78 (1H, m),

7.76 (1H, d, J = 7.5Hz), 7.66 (1H, t, J = 7.5Hz), 7.65 (1H, s),

7.55 (1H, d, J = 8.7Hz), 7.21 (1H, d, J = 8.7Hz).

FABMS (MH ^&lt; ^{+ &gt;} ): 381.

Anal. calcd for C ₁₇ H ₁₂ N ₆ O ₃ S: C, 53.68; H, 3.18; N, 22.09; WITH,

8.43. Found: C, 53.69; H, 3.14; N, 21.99; S, 8.39.

r e * Example A (10):

(4-Amino-2- (4-methoxy-phenylamino) -thiazol-5-yl) - (2-nitrophenyl) methanone

The title compound was prepared in a manner similar to that described in Example A (1). 4-Methoxyphenylisothiocyanate and 2-bromo-2'-nitroacetophenone gave, after recrystallization from aqueous ethanol, 562 mg (43% yield) of red-brown crystals, m.p. 185-188 ° C.

1 H NMR (DMSO-d ₆ ): δ 10.65 (1H, s), 8.25 (2H, bs), 8.20 (1H, d, J = 7.5Hz), 7.77 (IH, t, J = 7.5 Hz), 7.66 (1H, t, J = 7.5 Hz),

7.62 (1H, d, J = 7.5Hz), 7.41 (2H, d, J = 8.7Hz), 6.92 (2H, d,

J = 8.7 Hz), 3.72 (3H, s).

FABMS (M + Na &lt; + & gt ^; ): 393.

Anal. calcd for C ₁₇ H ₁₄ N ₄ O ₄ S: C, 55.13; H, 3.81; N, 15.13; S, 8.66. Found: C, 55.08; H, 3.83; N, 15.11; S, 8.56.

* Example A (11):

(4-Amino-2- (pyridin-3-ylamino) -thiazol-5-yl) - (2-nitrophenyl) methanone

The title compound was prepared essentially as in Example A (1). Pyridin-3-yl-isothiocyanate and 2-bromo-2'-nitroacetophenone gave, after column chromatography with 5% MeOH / CH ₂ Cl ₂ , rff-rrr erc rrrrr

Γ <yy yy yy: yy yy yy

750 mg (42% yield) of a yellow solid, m.p. 143.5-146.0 ° C.

1 H NMR (DMSO-d ₆ ): _δ 10.95 (1H, bs), 8.62 (1H, s), 8.19 (1H, dd, J = 4.7, 1.3 Hz), 8 , 08-7.86 (4H, m), 7.76 (1H, td, J = 8.3, 0.9 Hz), 7.66 (IH, dd, J = 8.4, 1.3 Hz) ), 7.62 (1H, d, J = 7.5Hz),

7.31 (1H, dd, J = 8.4, 4.7 Hz).

FABMS (MH ^&lt; ^{+ &gt;} ): 342.

Anal. calcd for C ₁₅ H _U N ₅ O ₃ S. 0.5 H ₂ O. 0.4 EtOH: C, 51.46; H, 3.94; N, 18.99; S, 8.69. Found: C, 51.86; H, 3.88; N, 19.24;

S, 8.88.

* Example A (12): Methyl acid ester

4-Amino-2- (1H-benzoimidazol-6-ylamino) -thiazole-5-carboxylic acid

The title compound was prepared in a manner similar to that described in Example A (1). 6-Isothiocyanate-1H-benzoimidazole (see Boev et al., Pharm. Chem. J. (English translation), vol. 24 (1990), pp. 812-822) and methyl bromoacetate gave a yellow solid in 63% yield, tt 266-267 [deg.] C.

1 H NMR (DMSO-d 6): δ 12.37 (1H, bs), 10.52 (IH, s), 7.92 (IH, s), 7.52 (IH, d, J = 8.7) Hz), 7.23 (1H, dd, J = 8.7, 1.9 Hz), 6.89 (2H, bs), 3.62 (3H, s).

FABMS (MH ^&lt; ^{+ &gt;} ): 250.

Anal. calculated for Ο ^ Η ^ Ο ^. 0.15 EtOH: C, 49.87; H, 4.05; N,

23.64; S, 10.82. Found: C, 49.94; H, 3.94; N, 23.41; S, 10.79.

Example A (13) r

prprrr furnace ρ i ^r ρ / Γ rrrrrrr RRR - r / 4-Amino-2- (p-tolylamino) -thiazol-5-yl / - (2, 4-dimethoxyphenyl) methanone

The title compound was prepared in a manner similar to that described in Example A (1). p-Tolylisothiocyanate and 2-bromo-2 ', 4'-dimethoxyacetopheno provided, after recrystallization from MeOH / CHCl ₃ , 78 mg (24% yield) of a yellow solid, mp 215-216 ° C.

1 H NMR (DMSO-d 6): δ 10.51 (1H, s), 7.88 (2H, bs), 7.41 (2H, d, J = 8.4 Hz), 7.16 (2H, d) J = 8.4 Hz), 7.12 (2H, d, J = 8.4 Hz),

6.58 (1H, d, J = 2.2 Hz), 6.52 (1H, dd, J = 8.4, 2.2 Hz), 3.78 (3H, s), 3.74 (3H) s, 2.24 (3H, s).

IR (KBr): 3279, 2959, 16.6, 1515, 1432, 1306, 1284, 1209, 1157, 1124, 1032 cm ^-1 .

Anal. calcd for C ₉ H ₁₉ N ₃ O ₃ S: C, 61.77; H, 5.18; N, 11.37; S, 8.68. Found: C, 61.69; H, 5.16; N, 11.33; S, 8.79.

* Example A (14):

(4-Amino-2- (p-tolylamino) -thiazol-5-yl) - (2,4-dimethylphenyl) methanone

The title compound was prepared in a manner similar to that described in Example A. (1). p-Tolylisothiocyanate a

2-bromo-2 ', 4'-dimethylacetophenone gave, after recrystallization from r r r r r r r r r r r r r r

P P p p • • P P p P P r. · P p c p

MeOH / CHCl ₃ , 65 mg (33% yield) of yellow crystals, mp 220-221 ° C.

¹ H NMR (DMSO-d ₆ ): _δ 10.58 (1H, s), 7.99 (2H, bs), 7.39 (2H, d, J = 8.1 Hz), 7.17 (2H δ, J = 7.8 Hz), 7.13 (2H, d, J = 8.1 Hz), 7.04 (IH, s), 7.00 (IH, d, J = 8.7 Hz) 1.73 (3-H, s), 2.24 (3H, s), 2.22 (3H, s).

IR (KBr): 3266, 2921, 1612, 1598, 1546, 1518, 1431 ^cm-first

Anal. calcd for C ₁₉ H ₁₉ N ₃ OS: C, 67.63; H, 5.68; N, 12.45; S, 9.50. Found: C, 67.70; H, 5.73; N, 12.47; S, 9.62.

* Example B:

(4-Amino-2- (p-tolylamino) -thiazole-5-carbonyl) -phenyl-benzoate

The intermediate, S- (4-benzoyloxyphenylacetyl) -N'-cyano-N-β-tolyl isourea, was first prepared according to the method of Gevalda et al., J. Prakt, Chem., Vol. 35 (1967), pp. 97-104. Sodium (6.7 mg, 0.29 mmol) was carefully dissolved in methanol (0.5 mL) and allowed to cool to ambient temperature. To the obtained solution was added p-tolylisothiocyanate (43 mg, 0.29 mmol) and cyanamide (12 g, 0.29 mmol). After 1 hour, 4-bromoacetylphenylbenzoate (92 mg, 0.29 mmol) was added and the resulting mixture was stirred overnight at ambient temperature. The mixture was then diluted with water (10 mL). Tanning substance obtained solid was filtered off, washed with water and a small amount of ether, dried under vacuum and recrystallized from ethanol / CHC1 ₃ to give a starting dose of 63 mg (51% yield) of S- (4-Benzoyloxyphenylacetyl) -Ν'- cyano-N ' -p-tolyl-isomers (as white needles):

r cr rrr rrrr rrcrr · R · _r f * rr - arrrr <* · r. · * Rr 'rr Γ Γ _r rrrc' ^X 'HNMR (DMSO-d _6): δ 8.10 to 8.04 (2H, m), 7.72 (1H, ddd, J = 7.5, 1 Δ, 1.8 Hz), 7.67 - 7.54 (4H, m), 7.20 (2H, d, J = 8.7 Hz), 7.03 (4H, dd, J = 11), 2.8 (Hz, d, J = 12.1 Hz), 3.77 (1H, d, J = 12.1 Hz), 2.19 (3H, s).

From the intermediate, the title compound was prepared as follows: Crude S- (4-benzoyloxyphenylacetyl) -N'-cyano-N '- p-tolyl-isourea (0.29 mmol) and triethylamine (101 μΐ, 0.73 mmol) ) in ethyl acetate (1 mL) was refluxed for 2 hours, then allowed to cool to ambient temperature, and concentrated in vacuo to a crude solid that crystallized from MeOH / CHCl ₃ in portions to give 67 mL (54% yield) of yellow needles, mp 245-247 ° C.

^Δ NMR (DMSO-d ₆ ): δ 10.71 (1H, s), 8.34-8.11 (4H, m), 7.80-7.72 (3H, m), 7.66- 7.57 (2H, m), 7.46 (2H, d, J = 8.1 Hz), 7.38 (2Ή, d, J = 8.4 Hz), 7.16 (2H, d, J = 8.4 Hz), 2.26 (3H, s).

IR (KBr): 3451, 3332, 3026, 1732, 1597, 1533, 1444, 1264, 1206, 1164, 1062, 708 cm &^lt; -1 & gt ^; .

HRFABMS: calcd for C ₂₄ H ₁₉ N ₃ O ₃ SC 5 (M + Cs 2: 562.0201. Found: 562.0184).

Anal. Calcd for C ₂₄ H ₁₉ N ₃ O ₃ S: C, 67.12; H, 4.46; N, 9.78; WITH,

7.47. Found: C, 66.90; H, 4.43; N, 9.70; S, 7.50.

* Example C (1): Methyl acid ester

4- / 4-amino-5- (4-methoxy-benzoyl) -thiazol-2-ylamino / -benzoic what · h ₃

ih ₂ Of their ₃ yy

To a mixture of 4-methoxycarbonylphenylisothiocyanate (82 mg, 0.5 mmol) and cyanamide (23 mg, 0.55 mmol) in acetonitrile (5 mL) was added a solution of potassium tert-butoxide (61 mg, 0.55 mmol) in tert. butanol (5 mL). After 30 minutes at room temperature, 2-bromo-4'-methoxyacetophenone (115 mg, 0.5 mmol) was added. After 2 hours at ambient temperature, the reaction mixture was diluted with water (50 mL). The product was collected by filtration, washed with water and ethyl ether, and dried under vacuum to give a yellow solid, 172 g (90% yield).

¹ H NMR (DMSO-d ₆ ): δ 8.00 (2H, d, J = 8.2 Hz), 7.84 (2H, d, J = 8.2 Hz), 7.72 (2H, d, J = 8.2 Hz), 7.10 (2H, d, J = 8.2 Hz), 3.90 (6H, s).

FABMS (MH &lt; + &gt;): 384

Anal. calculated for C ₁₉ H ₁₇ N ₃ O ₄ S. 0.5 H ₂ O: C, 57.36; H, 4.71; N, 10.56; S, 8.06. Found: C, 56.97; H, 4.74; N, 10.51; S, 8.07.

* Example (2):

(4-Amino-2- (4-benzyloxy-phenylamino) -thiazol-5-yl) - (4-methoxy-phenyl) -methanone

The title compound was prepared in a similar manner to that described in Example C (1). 4-Benzyloxyphenylisothiocyanate and 2-bromo-4'-methoxyacetophenone gave a yellow-brown solid in 85% yield, m.p. Mp 222-224 ° C.

¹ H NMR (DMSO-d 6): δ 7.70 (2H, d, J = 8.2 Hz), 7.58-7.34 (7H, m), 7.06 (4H, dd, J = 7, Δ, 1.2 Hz), 5.18 (2H, s), 3.94 (3H, s).

FABMS (MH &lt; + &gt;): 432

Anal. calculated for C ₂₄ H ₂₁ N ₃ O ₃ S: C, 66.80; H, 4.91; N, 9.47; S, 7.43. Found: C, 66.86; H, 4.91; N, 9.76; S, 7.53.

c P r * r

P P r P * Example C (3):

(4-Amino-2- (4-dimethylamino-phenylamino) -thiazol-5-yl) - (4-methoxyphenyl) methanone

The title compound was prepared in a manner similar to that described in Example C (1) from dimethylamino-phenylisothiocyanate and 2-bromo-4'-methoxyacetophenone to give the product as a yellow solid in 85% yield, m.p. 178-180 ° C.

¹ H NMR (DMSO-d 6): δ 7.70 (2H, d, J = 8.2 Hz), 7.34 (2H, d, J = 8.2 Hz), 7.00 (2H, d, J = 8.6 Hz), 6.80 (2H, d, J = 8.6), 3.94 (3H, s), 2.94 (6H, s).

Anal. Calcd. for C ₁₉ H ₂₀₁ N ₄ O ₂ S: C, 61.94; H, 5.47; N, 15.21; S, 8.70. Found: C, 66.22; H, 5.48; N, 15.03; S, 8.58.

* Example C (4):

(4-Amino-2- (4-dimethylamino-phenylamino) -thiazol-5-yl) - (2-nitrophenyl) -methanone

The title compound was prepared in a similar manner to that described in Example C (1). 4-Dimethylaminophenylisothiocyanate and 2-bromo-2'-nitro-acetophenone gave a yellow solid in 90% yield, m.p. greater than 195 ° C (dec.).

¹ H NMR (DMSO-d ₆ ): δ 8.06 (2H, bs), 7.76 (2H, m), 7.27 (2H, bs), 6.74 (2H, d, J = 9), 0 Hz), 3.38 (6H, s).

FABMS (MH ^&lt; ^{+ &gt;} ): 384.

Anal. calculated for Ο _1Θ Η ₁₇ Ν ₅ Ο ₃ 8. 0.5 CH ₃ CN: C, 56.94; H, 4.62; N,

19.07; S, 7.94. Found: C, 56.71; H, 4.64; N, 19.09; S, 7.88.

* Example C (5):

(4-Amino-2-phenylethylamino-thiazol-5-yl) - (2-nitrophenyl) methanone

The title compound was prepared essentially similar to that described in Example C (1). Phenethylisothiocyanate and 2-bromo-2'-nitroacetophenone gave an amorphous yellow solid in 90% yield, m.p. 75.0-81.5 ° C (dec.).

NMR (DMSO-d _6): δ 8.67 (1H, bs), 8.00 (1H, d, J = 8.1 Hz), 7.80

(2H, bs) 7.75 (1H, t, J = 7.5Hz), 7.65 (1H, t, J = 7.5Hz), 7.58 (1H, d, J = 6.5 Hz), 7.04 7.32 (5H, m), 3.50 (2H, bs), 2.82 (2H, t, J = 7.2 Hz).

FABMS (MH &lt; + &gt;): 369.

Anal. calcd for C ₁₆ H _ig N ₄ O ₃ S. 0.1 H ₂ O. 0.1 C ₆ H ₁₄ : C, 58.97; H, 4.68; N, 14.79; S, 8.46. Found: C, 58.97; H, 4.78; N, 14.54; S, 8.37.

* Example C (6):. Methyl 2 (S) - [4-Amino-5- (4-nitro-benzoyl) -thiazol-2-ylamino] -butyrate

The title compound was prepared in a similar manner to that described in Example C (1). Methyl 2 (S) -isothiocyanato-butyrate and 2-bromo-4'-nitroacetophenone gave an amorphous red-brown solid in 89% yield.

¹ H NMR (CDCl 3): δ 8.28 (2H, d, J = 8.2 Hz), 7.86 (2H, J = 8.2 Hz), 3.94 (3H, s), 4.32 (1H, bs), 2.12 (1H, m), 1.88 (1H, m), 0.96 (3H, t, J = 6.4 Hz).

I * c * C

Γ r Γ <· ^r er er 'r 9

FABMS (MH &lt; + &gt;): 365 * Example C (7):

[4-Amino-2 - ((4-dimethylaminophenyl) amino) thiazol-5-yl] - (3-methylbenzo [b] thiophen-2-yl) methanone

The title compound was prepared essentially as described in Example C (1). 4-Dimethylminophenylisothiocyanate and 2- (2-bromoacetyl) -3-methylbenzo [b] thiophene gave, after recrystallization from ethanol, 210 mg (92% yield) of a yellow powder, m.p. Mp 123-126 ° C.

¹ H NMR (DMSO-d 6): δ 10.50 (1H, s), 8.20 (2H, bs), 7.96 (1H, ddd, J = 5.0, 5.0, 1.9 Hz) 7.82 (1H, ddd, J = 4.1, 4.1, 1.7), 7.44 (2H, ddd, J = 9.0, 4.5, 4.5 Hz), 7 26 (2H, d, J = 8.5 Hz), 6.69 (2H, d, J = 9.0 Hz), 2.84 (6H, s).

FABMS (MH &lt; + &gt;): 409.

Anal. calculated for C ₂₁ H ₂₀ N ₄ OS ₂ . 0.5 H ₂ 0. 0.5 MeOH: C, 59.56; H, 5.35; N, 12.92; S, 14.79. Found: C, 59.87; H, 5.39; N, 12.86;

S, 14.69.

* Example (8):

/ ^R 4-Amino-2- (lH-benzoimidazol-6-ylamino) -thiazol-o-yl / - (3-methylbenzo / b7thiofen-2-yl) -methanone

The title compound was prepared similarly to Example C (1). 6-Isothiocyanate-1H-benzoimidazole (see Boev et al., Pharm. Chem. J. (Eng. Trans.)., Vol. 24 (1990), pp. 818-822) and 2- (2-bromoacetyl) - 3-methyl-benzo [b] thiophene gave 160 mg (53% yield) of a yellow powder, m.p. Mp 235-240 ° C.

¹ H NMR (DMSO-d ₆ ): δ 12.50 (1H, s), 10.9 (IH, s), 8.28 (2H, bs), 8.19 (IH, s), 8.20 -7.93 (1H, m), 7.93-8.00 (2H, m), 7.56 (IH, d, J = 8.7 Hz), 7.50-7.40 (2H, m) 7.25 (1H, d, J = 8.7 Hz), 2.49 (3H, s).

FABMS (MH &lt; + &gt;): 406.

Anal. calculated for C ₂₀ H ₁₅ N ₅ OS ₂ . 0.5 H ₂ O: C, 57, 95; H, 3.89; N, 16.90; S, 15.47. Found: C, 57.98; H, 3.88; N, 17.06; S, 15.55. * Example (9):

4-Amino-2- (5-chloro-3-methyl-benzo [b] thiophen-2-ylamino) -thiazol-5-yl] - (4-dimethylamino-phenyl) -methanone

The title compound was prepared essentially as in Example C (1). 4-Dimethylaminophenyl isothiocyanate and 2- (2-bromoacetyl) -5-chloro-3-methylbenzo [b] thiophene gave 54% yield of a yellow powder, m.p. 265-268 [deg.] C.

¹ H NMR (DMSO-d ₆ ): δ 10.60 (1H, s), 8.04 (2H, bs), 8.00 (IH, d, J =

8.7 Hz), 7.88 (1H, d, J = 1.8Hz), 7.46 (IH, dd, J = 8.7, 1.8Hz), 7.34-7.20 ( 2H, m), 6.68 (2H, d, J = 9.0 Hz), 2.85 (6H, s), 2.43 (3H, s).

FABMS (MH &lt; + &gt;): 443/445.

Anal. Calcd for C ₂₁ H ₁₉ N ₄ OS ₂ Cl: C, 56.94; H, 4.32; N, 12.65; S, 14.48; Cl, 8.00. Found: C, 56.82; H, 4.39; N, 12.42; S, 14.42; Cl, 8.17.

* Example C (10):

4-Amino-2- (1H-benzimidazol-6-yl-amino) -thiazol-5-yl-N- (5-chloro-3-

-methyl-benzo [b] thiophen-2-yl) -methanone r

The title compound was prepared in analogy to Example C (1). 6-Isothiocyanato-1H-benzoimidazole (see Boev et al., Pharm. Chem. J. (Eng. Trans.)., Vol. 24 (1990), pp. 818-822) and 2- (2-bromoacetyl) -5- chloro-3-methyl-benzo [b] thiophene gave a 59% yield of a yellow powder, m.p. 275-280 ° C.

Ή NMR (DMSO-d ₆ ): δ 12.44 (1H, s), 10.86 (1H, s), 8.30 (2H, bs), 8.18 (1H, s), 8.02 ( 1H, d, J = 8.4Hz), 7.90 (1H, d, J = 2.0Hz), 7.86 (1H, bs), 7.55 (1H, d, J = 8.4 Hz), 7.45 (1H, dd, J = 8.7, 2.0 Hz), 7.25 (1H, d, J = 8.7 Hz), 2.46 (3H, s).

ASIMS (MH &lt; + &gt;): 440/442.

Anal. calculated for C ₂₀ H ₁₄ N ₅ OS ₂ C1. H, O: C, 52.45; H, 3.52; N, 15.29; S, 14.00; Cl, 7.74. Found: C, 52.61; H, 3.60; N, 15.15; S, 14.12; Cl, 7.81.

* Example C (11):

(4-7) mino-2- (benzo [1,3] dioxol-5-yl-amino) -thiazol-5-yl? ^- (2-nitrophenyl) methanone

NH,

NOj

The title compound was prepared in analogy to Example C (1). 3,4-methylenedioxyphenyl-isothiocyanate and 2-bromo-2'-nitro-acetophenone gave a yellow solid in 73% yield, m.p. 200.0 - 202.5 ° C.

Ή NMR (DMSO-d 6): δ 10.69 (1H, s), 8.04 (2H, bs), 8.03 (1H, d, J =

7.8 Hz), 7.78 (1H, dd, J = 7.8, 6.5 Hz), 7.67 (1H, dd, J = 7.2,

6.5 Hz), 7.63 (1 H, d, J = 7.2 Hz), 7.28 (1 H, s), 6.89 (1 H, d, J =

8.9 Hz), 6.85 (1H, d, J = 8.9 Hz), 6.00 (2H, s).

FABMS (MH &lt; + &gt;): 385.

C c c

C r ·

Anal. calcd for C ₁₇ H ₁₂ N ₄ O ₅ S: C, 53.12; H, 3.15; N, 14.58; S, 8.34. Found: C, 53.02; H, 3.20; N, 14.39; S, 8.27.

* Example C (12):

(4-Amino-2- (4-methoxy-phenylamino) -thiazol-5-yl) - (2-iodo-phenyl) -methanone

2-Bromo-2'-iodoacetophenone having the structural formula was first prepared as follows. Following the procedure of King et al., J. Org. Chem. 29 (1964), pages 3459-3461, was added copper bromide (6.34 g, 28.8 mmol) to a solution of 2'-iodoacetophenone (3.54 g, 14.4 mmol) in EtOAc, and the resulting mixture was heated under reflux for 90 minutes. The mixture was allowed to cool and the solids were filtered and washed with EtOAc. The filtrate was concentrated to give 4.60 g (98% yield)

2-bromo-2'-iodoacetophenone as a yellow oil, which was consistent with the previously described material (Lutz et al., J. Org. Chem., Vol. 12 (1947), p. 617).

The title compound was then prepared essentially as described in Example C (1). 4-Methoxyphenylisothiocyanate and 2-bromo-2'-iodoacetophenone gave a yellow solid in 71% yield, m.p. 187-190 ° C.

¹ H NMR (DMSO-d ₆ ): δ 10.56 (1H, s), 8.03 (2H, bs), 7.85 (1H, d, J =

7.5 Hz, 7.32-7.48 (3H, m), 7.29 (1H, dd, J = 7.5, 1.6 Hz), 7.12 (1H, td, J = 7) 1.5, 1.6 Hz), 7.12 (1H, td, J = 7.5, 1.6 Hz), 6.90 (2H, d, J = 9.0 Hz), 3.51 (3H) , s).

FABMS (MH &lt; + &gt;): 452.

Anal. calculated for C ₁₇ H ₁₄ N ₃ O ₂ S ₁ · 0.05 C ₆ H ₆ . 0.2 EtOH: C, 45.78; H,

3.36; N, 9.05; S, 6.90; I, 27.53. Found: C, 46.08; H, 3.54; N, r f βe

9.09; S, 7.04; I, 27.62.

* Example C (13):

(4-Amino-2- (4-nitro-phenylamino) -thiazol-4-yl) - (2-nitro-phenyl) -methanone

The title compound was prepared as described in Example C (1). 4-Nitrophenylisothiocyanate and 2-bromo-2'-nitroacetophenone gave a yellow solid in 45% yield, m.p. 266.0-268.2 ° C.

NMR (DMSO-d _6): δ 10.80 (IH, s), 8.23 (2H, d, J = 9.4 Hz), 8.15 (2H, bs), 8.08 (IH, d J = 8.1 Hz) 7.84 (2H, d, J = 9.4 Hz),

7.83 (1H, t, J = 7.5Hz), 7.75-7.66 (2H, m).

FABMS (MH &lt; + &gt;): 386.

Anal. Calcd for C 18 H 18 N 8 O 5 S: C, 49.87; H, 2.88; N, 18.17; S, 8.32. Found: C, 49.83; H, 2.90; N, 18.10; S, 8.27.

* Example C (14):

(4-Amino-2-cyclohexylamino-thiazol-5-yl) - (2-nitro-phenyl) -methanone

The title compound was prepared in analogy to Example C (1). Cyclohexylisothiocyanate and 2-bromo-2'-nitroacetophenone gave a yellow solid in 45% yield, m.p. Mp 116-118 ° C.

NMR (DMSO-d6): δ 8.62 (1H, bs), 8.00 (IH, d, J = 8.1 Hz), 7.97 (2H, bs), 7.75 (1H, dd, J = 8.1, 7.5 Hz), 7.64 (1H, dd, J = 8.1,

7.5 Hz), 7.59 (1H, d, J = 7.5Hz), 3.62 (IH, bs), 1.94-1.78 (2H, m), 1.73-1, 60 (2H, m), 1.58-1.46 (1H, m), 1.32-1.02 (5H, m).

Ρ Ρ

FABMS (+): 347.

Anal. Calcd for C ₁₆ H ₁₈ N ₄ O ₃ S · 0.7 H ₂ O: C, 53.53; H, 5.45; N, 15.61; S, 8.93. Found: C, 53.79; H, 5.24; N, 15.44; S, 8.93.

* Example C (15):

(4-Amino-2- (4-isopropyl-phenylamino) -thiazol-5-yl) - (2-nitro-phenyl) -methanone

no ₂

The title compound was prepared essentially as described in Example C (1). Isopropyl isothiocyanate and 2-bromo-2'-nitroacetophenone gave a yellow solid in 58% yield, m.p. 202.5-205.0 ° C.

^L H NMR (DMSO-d _s): δ 10.74 (H, s), 8.05 (2H, bs), 8.03 (H, d, J =

7.5 Hz), 7.78 (1H, dt, J = 7.5, 1.3 Hz), 7.71-7.60 (2H, m),

7.41 (2H, d, J = 8.3 Hz), 7.20 (2H, d, J = 8.3 Hz), 2.83 (1H, heptet, J = 6.9 Hz), 1, 16 (6H, d, J = 6.9Hz).

FABMS (MH &lt; + &gt;): 383.

Anal. calcd for C ₁₉ H ₁₈ N ₄ O ₃ S: C, 59.67; H, 4.74; N, 14.65; S, 8.38. Found: C, 59.62; H, 4.77; N, 14.56; S, 8.43.

* Example C (16):

// 1-Amino-2- [2- (4-chlorophenyl) -ethylamino-thiazol-5-yl] - (2-nitrophenyl) -methanone

The title compound was prepared in a manner similar to that described in Example C (1). 4-Chlorophenethylisothiocyanate and 2-bromo-2'-nitroacetophenone gave a yellow solid in 61% yield, m.p. Mp 117-120 ° C.

rercrec ^T H NMR (DMSO-d _6): δ 8.74 (1H, bs), 8.00 (1H, d, J = 8.1 Hz), 7.95 (2H, bs), 7.75 ( 1H, td, J = 7.5, 1.2 Hz), 7.64 (2H, td, J = 8.1,

1.6 Hz), 7.57 (1H, dd, J = 7.5, 1.2 Hz), 7.33 (2H, d, J = 8.4 Hz), 7.23 (2H, d, J = 8.4 Hz), 3.60-3.35 (2H, m), 2.81 (2H, t, J = 6.8 Hz).

FABMS (MH &lt; + &gt;): 403.

Anal. Calcd. for C ₁₃ H ₁₅ N ₄ O ₃ SC 1.0, 5 EtOH: C, 53.58; H, 4.26; N, 13.16; S, 7.53; Cl, 8.32. Found: C, 53.63; H, 4.33; N, 13.22; S, 7.47; Cl, 8.45.

* Example C (17):

4-Amino- (2- (4-diethylamino-phenylamino) -thiazol-5-yl) - (2-nitrophenyl) -methanone

The title compound was prepared in a manner similar to that described in Example C (1). 4-Diethylaminophenyl isothiocyanate and 2-bromo-2'-nitroacetophenone gave a yellow solid in 63% yield, m.p. 202.5-205.0 ° C.

¹ H NMR (DMSO-d ₆ ): δ 10.45 (1H, s), 8.01 (1H, d, J = -8.1 Hz), 7.97 (2H, bs), 7.75 ( 1H, dd, J = 8.1, 7.8 Hz), 7.64 (1H, dd, J = 8.1, 7.8 Hz), 7.59 (1H, d, J = 7.8 Hz) 7.18 (2H, d, J = 9.0 Hz),

6.61 (2H, d, J = 9.0 Hz), 3.28 (4H, q, J = 7.2 Hz), 1.05 (6H, t, J = 7.2 Hz).

FABMS (MH &lt; + &gt;): 412.

Anal. Calcd. for C ₂₀ H ₂₁ N ₅ O ₃ S: C, 58.38; H, 5.14; N, 17.02; S, 7.79. Found: C, 58.28; H, 5.20; N, 16.77; S, 7.94.

* Example C (18):

(4-Amino-2- (4-diethylamino-phenylamino) -thiazol-5-yl) - (4-nitrophenyl) -methanone

The title compound was prepared in a manner similar to that described in Example C (1). 4-Diethylaminophenyl isothiocyanate and 2-bromo-4'-nitroacetophenone gave a yellow solid in 63% yield, m.p. Mp 220-221 ° C.

¹ H NMR (DMSO-d ₆ ): δ 10.51 (1H, s), 8.42 (2H, bs), 8.26 (2H, d, J = 12.0 Hz), 7.84 (2H δ d, J = 12.0 Hz), 7.22 (2H, d, J = 9.0 Hz), 6.63 (2H, d, J = 9.0 Hz), 3.26 (4H, q J = 6.8 Hz) 1.05 (6H, t,

J = 6.8 Hz).

FABMS (MH &lt; + &gt;): 412.

Anal. Calcd. for C ₂₀ H ₂₁ N ₅ O ₃ S: C, 58.38; H, 5.14; N, 17.02; S, 7.79. Found: C, 58.23; H, 5.16; N, 16.94; S, 7.86.

* Example C (19):

/ 4-Amino-2- (lH-benzoimidazol-6-ylamino) -thiazol-5-yl / - (3-methyl-thiophen-2-yl) -methanone NH ₂

The title compound was prepared in a manner similar to that described in Example C (1). 6-Isothiocyanate-1H-benzoimidazole (see Boev et al., Pharm. Chem. J. (English trans.)., Vol. 24 (1990), pp. 812-822) and 2- (2-bromoacetyl) - 3-methylthiophene (U.S. Pat. No. 5,189,049; acetyl brominated by copper bromide according to the method of Kong et al., J. Org. Chem., Vol. 29 (1964), pp. 3459-3461; exemplary procedure in Example C (19) )) gave a yellow powder in 67% yield, mp 285-287 ° C.

eree ^X H NMR (DMSO-d _6): δ 12.60 (1H, bs), 10.78 (1H, s), 8.23 (1H, s), 8.17 (2H, bs), 7, 93 (1 H, s), 7.56 (1 H, d, J = 8.7 Hz), 7.55 (1 H, d, J = 5.0 Hz), 7.27 (1 H, dd, J = 8) 7. 1.9 (Hz), 6.60 (1H, d, J = 5.0 Hz), 2.36 (3H, s).

FABMS (MH &lt; + &gt;): 356.

Anal. Calcd. for C ₁₅ H ₁₃ N ₅ OS ₂ .0.6 H ₂ O: C, 52.47; H, 3.91; N,

19.12; S, 17.51. Found: C, 52.50; H, 3.90; N, 19.10; S, 17.71. * Example C (20) [4-Amino-2- (1H-benzoimidazol-6-ylamino) -thiazol-5-yl] - (2,4-dimethylphenyl) methanone

The title compound was prepared in a manner similar to that described in Example C (1j, 6-Isothiocyanate-1H-benzoimidazole (see Boev et al., Pharm. Chem. J., vol. 24) (1990). 812-822) and 2-bromo-2 ', 4'-dimethylacetophenone gave a 77% yield of a yellow powder, mp 290-292 ° C.

^X H NMR (DMSO-d _s): δ 12.43 (1H, bs), 10.65 (1H, s), 8.18 (2H, bs), 7.80 (1H, bs), 7.54 (1H, d, J = 8.7Hz), 7.20 (1H, d, J = 8.7Hz), 7.16 (1H, d, J = 7.5Hz), 7.03 (1H) s), 6.99 (1H, d, J =

7.5 Hz), 2.26 (3H, s), 2.22 (3H, s).

FABMS (MH &lt; + &gt;): 364.

Anal. calcd for C ₁₉ H ₁₇ N ₅ OS: C, 62.79; H, 4.71; N, 19.27; WITH,

8.82. Found: C, 62.50; H, 4.78; N, 19.22; S, 8.72.

* Example C (21) 4-Amino-2- (pyridin-3-ylamino) -thiazol-5-yl- (2,4-dimethylphenyl) methanone

o ρ ρ e r

The title compound was prepared in a manner similar to that described in Example C (1). 3-Pyridyl isothiocyanate a

2-bromo-2 ', 4'-dimethylacetophenone gave a 63% yield of a yellow powder, m.p. 200-202 ° C.

Ή NMR (DMSO-d ₆ ): δ 10.82 (1H, s), 8.76 (1H, d, J = 2.5Hz), 8.06 (1H, d, J = 8.4 Η) 8.04 (2H, bs), 7.36 (1H, dd, J = 8.4,

4.1 Hz), 7.21 (1H, d, J = 7.5Hz), 7.06 (1H, s), 7.02 (1H, d, J = 7.5Hz), 2.28 (2H, s) 2.23 (3H, s).

FABMS (MH &lt; + &gt;): 325.

Anal. Calcd. for C ₁₇ H ₁₆ N ₄ OS: C, 62.94; H, 4.97; N, 17.27; WITH,

9.88. Found: C, 65.86; H, 5.03; N, 17.17; S, 9.95.

* Example C (22)

3- / 4-Amino-5- (2-cyano-benzoyl) -thiazol-2-ylamino-7-benzonitrile

The title compound was prepared essentially as described in Example C (1). 3-Cyanophenyl isothiocyanate and 2-bromo-2'-nitroacetophenone gave an orange solid in 94% yield, m.p. Mp 235-236 ° C.

¹ H NMR (DMSO-d ₆ ): δ 8.26 (1H, bs), 8.06 (1H, d, J = 8.0 Hz), 7.8 (1H, t, J = 7.0 Hz) 7.74-7.64 (3H, m), 7.58-7.48 (2H, m).

IR (KBr): 3460, 33.7, 3271, 3083, 2214, 1625, 1601, 1525 cm ^-1 .

Anal. calcd for C ₁₇ H _U N ₅ O ₃ S: C, 55.80; H, 3.03; N, 19.17; S, 8.78. Found: C, 55.70; H, 3.05; N, 19.01; S, 8.73.

* Example C (23) [4-Amino-2- (3-methoxypropylaxnino) -thiazol-5-yl] - (2-nitrophenyl) methanone

-A mh ₂

_NO2 H ₃ coSloučenina The title compound was prepared in a manner similar to that described for Example C (l). 3-Methoxypropyl isothiocyanate and 2-bromo-2'-nitroacetophenone gave a yellow solid in 99% yield, mp 170-172 ° C.

¹ H NMR (DMSO-d 6): δ 8.02-7.92 (2H, m), 7.4 (1H, t, J = 7.0 Hz), 7.68-7.56 (2H, m) 3.38-3.22 (7H, m), 1.78-1.66 (2H, m).

Anal. Calcd. for C ₁₄ H ₁₆ N ₄ O ₄ S: C, 49.99; H, 4.79; N, 16.66; S, 9.53. Found: C, 50.04; H, 4.81; N, 16.69; S, 9.61.

* Example C (24) I ^- <4-Amino-5- (2-nitro-benzoyl) -thiazol-2-ylaminq / fenylJethanon

The title compound was prepared in a manner similar to that described in Example C (1). 4-Acetylphenyl isothiocyanate and 2-bromo-2'-nitroacetophenone gave a yellow solid in 87% yield, m.p. 264-265 [deg.] C.

¹ H NMR (DMSO-d ₆ ): δ 8.06 (1H, d, J = 8.0 Hz), 7.92 (2H, d, J = 9.0 Hz), 7.84. 7.78 (1H, m), 7.73-7.64 (4H, m), 2.42 (3H, s).

IR (KBr): 3389, 3248, 1690, 1655, 1573, 1472, 1420, 1273 cm &^lt; -1 & gt ^; .

Anal. calcd for C ₁₈ H ₁₄ N ₄ O ₄ S: C, 56.54; H, 3.69; N, 14.65; S, 8.39. Found: C, 56.39; H, 3.73; N, 14.44; S, 8.31.

* Example C (25) (4-Amino-2- [1- (2-chloro-5-trifluoromethyl-pyridin-2-ylsulfanyl) -phenylamino] -thiazol-5-yl] - (2-nitro-phenyl) -methanone

Γ Ρ

Ρ Ρ

Ρ Ρ * β ρ ρ ρ ρ ρ ρ γ ρ ρ ρ ρ ρ © ι- Ρ

T Ρ ρ ρ ρ ·, · C t Ρ f Γ

Ρ Γ. Γ

The title compound was prepared in a manner similar to that described in Example C (1).

2- [4- (2-Chloro-4-trifluoromethyl-pyridin-2-yl-sulfanyl) -phenyl] -isothiocyanate and 2-bromo-2'-nitroacetophenone gave an orange solid in 52% yield, m.p. 150-152 ° C.

NMR (DMSO-d6): δ 8.65 (1H, bs), 8.38 (1H, bs), 8.06 (2H, d, J = 8.0 Hz), 7.80 (1H, t, J = 7.0 Hz), 7.74-7.64 (4H, m), 7.54 (2H, d, J = 8.0 Hz).

IR (KBr): 3272, 3048, 1596, 1531, 1431, 1320 ^cm-first

Anal. Calcd for C ₂₂ H ₁₃ ClF ₃ N ₅ O ₃ S: C, 47.87; H, 2.37; N, 12.60; S, 11.62; Cl, 6.42. Found: C, 47.79; H, 2.44; N, 12.54; S, 11.70; Cl, 6.52.

* Example C (26)

Methyl 3- / 4-Amino-5- (2-methoxybenzoyl) -thiazol-2-ylamino / benzoate

The title compound was prepared in a manner similar to that described in Example C (1). 3-Methoxycarbonylphenylisothiocyanate and 2-bromo-2'-methoxy-acetophenone gave an ivory solid in 59% yield, m.p. Mp 214-215 ° C.

¹ H NMR (DMSO-d ₆ ): δ 10.81 (1H, s), 8.12-7.90 (4H, m), 7.62 (1H, ddd, J = 7.8, 1.2) 1.2 Hz), 7.49 (1H, t, J = 7.9 Hz), 7.39 (1H, ddd, J = 8.7, 8.7, 1.7 Hz), 7.25 (1H, dd, J = 7.5, 1.9 Hz), 7.09 (1H, d, J = 8.4 Hz), 6.98 (1H, ddd, J = 7.5, 7.5) 0.68 Hz, 3.85 (3H, s), 3.87 (3H, s).

r r e- r

1417, 1294

10.96; S, rr ^{ rrrf r-t- Λ c <

<- fr ·? rc ^r rcrrrr · 46

FABMS (MH &lt; + &gt;): 327.

IR (KBr): 3473, 3333, 3261, 3092, 1718, 1602, 1527, cm &lt; ^{-1 &gt};.

Anal. calcd for C ₁₉ H ₁₇ N ₃ O "S: C, 59.52; H, 4.47; N,

8.36. Found: C, 59.41; H, 4.46; N, 10.93; S, 8.3:

* Example C (27) [4-Amino-2- / 2- (4-chlorophenyl) -ethylamino-5-thiazol-5-yl] - (2-methoxy-phenyl) -methanone

The title compound was prepared in a manner similar to that described in Example C (1). The product 4-chlorophenethyl isothiocyanate and 2-bromo-2'-methoxyacetophenone was extracted with 10% i-PrOH / CHCl ₃ . The obtained solid was washed with Et ₂ O to give an ivory solid in 49% yield, mp 150-151 ° C.

1 H NMR (DMSO-d 6): δ 8.53 (2H, bs), 7.87 (1H, bs), 7.39-7.28 (3H, m), 7.23 (2H, d, J) = 8.4 Hz), 7.17 (1H, dd, J = 7.5, 1.6 Hz), 7.03 (IH, d, J = 8.4 Hz), 6.93 (IH, t , J = 7.5Hz),

3.40 (2H, bs), 2.81 (2H, t, J = 7.0 Hz).

FABMS (MH &lt; + &gt;): 388

IR (KBr) 3354, 3214, 3166, 3103, 1600, 1578, 1544,

1363 cm ~ ^s.

Anal. Calcd. for C ₁₉ H ₁₈ ClN ₃ O ₂ S: C, 58.83; H, 4.68;

8.27. Found: C, 58.70; H, 4.62; N, 10.75; S, 8.25.

* Example C (28) [4-Amino-2- (pyridin-3-ylamino) -thiazol-5-yl] - (2,4-chloro-phenyl) 3.88 (3H, s),

1525, 1462

N, 10.83; WITH,

-methanone ee e

O

The title compound was prepared in a manner similar to that described in Example C (1). 3-Pyridyl isothiocyanate and 2,2 ', 4'-trichloroacetophenone gave a yellow solid in 39% yield, m.p. Mp 209-210 ° C.

¹ H NMR (DMSO-d ₆ ): δ 10.95 (1H, s), 8.77 (1H, d, J = 2.5 Hz), 8.28 (1H, dd, J = 4.7, 1.6 Hz), 8.16 (2H, bs), 8.06 (1H, bd, J = 9.6 Hz), 7.70 (1H, d, J = 1.6 Hz), 7.48 (2H, dd, J = 11.5, 8.1 Hz), 7.37 (1H, dd, J = 8.4, 4.7 Hz).

FABMS (MH &lt; + &gt;): 365

IR (KBr): 3378, 3272, 3175, 3072, 1608, 1586, 1561, 1525, 1424 cm &^lt; -1 ^&gt;.

Anal. calculated for C ₁₅ H ₁₀ Cl ₂ N ₄ OS. 0.9 H ₂ O: C, 47.23; H, 3.12; N, 14.69; S, 8.41. Found: C, 47.03; H, 3.09; N, 14.52; S, 8.42.

* Example C (29) / * 4-Amino-2- (pyridin-3-ylamino) -thiazol-5-yl-2- (2-methoxy-phenyl) -methanone

The title compound was prepared in a manner similar to that described in Example C (1). 3-Pyridyl isothiocyanate and 2-bromo-2'-methoxyacetophenone gave an off-white / ivory solid in 67% yield, m.p. Mp 245-246 ° C.

¹ H NMR (DMSO-d ₅ ): δ 10.80 (1H, s), 8.77 (1H, d, J = 2.8 Hz), 8.25 (1H, dd, J = 4.7, 1.2 Hz), 8.07 (1H, ddd, J = 8.4, 2.8, 1.6 Hz),

8.00 (2H, bs), 7.44-7.33 (2H, m), 7.24 (1H, dd, J = 7.5, 1.6)

C ρ r r r r ρ f r Γ γ Γ ι Ρ

Γ Ρ

Δ), 7.09 (1H, d, J = 8.1 Hz), 6.98 (1H, t, J = 7.5 Hz), 3.76 (3H, s).

FABMS (MH &lt; + &gt;): 327

IR (KBr): 3424, 3310, 2971, 1632, 1603, 1526, 1459, 1405 cm.

Anal. Calcd. for C ₁₆ H ₁₄ N ₄ O ₂ S: C, 58.88; H, 4.32; N, 17.17; S, 9.28. Found: C, 58.84; H, 4.33; N, 17.07; S, 9.90.

Example C (30) / 4-Amino-2- (pyridin-3-ylamino) -thiazol-5-yl-naphthalen-2-ylmethanone

The title compound was prepared essentially as described in Example C (1). 3-Pyridyl isothiocyanate and 2-bromo-2'-acetophenone gave, after recrystallization from EtOH, a yellow solid in 12% yield, m.p. Mp 242-243 ° C.

¹ H NMR (DMSO-d 6): δ 10.97 (1H, s), 8.82 (1H, d, J = 2.5 Hz), 8.36 8.18 (3H, m), 8.13 ( 1H, ddd, J = 8.4, 4.0, 1.6 Hz), 8.08-7.93 (2H, m), 7.77 (IH, dd, J = 8.4, 1.6) Hz), 7.60 (2H, ddd, J = 14.3, 10.6, 7.9, 2.2 Hz), 7.39 (1H, dd, J = 8.4, 5.0 Hz) .

FABMS (MH &lt; + &gt;): 347.

IR (KBr): 3462, 3316, 3261, 3071, 1623, 1584, 1531, 1421 cm @ -1

Anal. calcd for C ₁₉ H ₁₄ N ₄ OS: C, 65.88; H, 4.07; N, 16.17; WITH,

9.26. Found: C, 65.80; H, 4.09; N, 16.09; S, 9.34.

* Example C (31) [4-Amino-2- (2-methoxy-benzylamino) -thiazol-5-yl] - (5-chloro-benzofuran-2-yl) -methanone

• ci yy yy *

The title compound was prepared in a manner similar to that described in Example C (1). 2-Methoxybenzyl isothiocyanate a

2-bromoacetyl-5-chlorobenzofuran gave a 62% yield of a yellow powder, m.p. Mp 241-242 ° C. .

1 H NMR (DMSO-d ₆ ): δ 9.17 (1H, bs), 8.78 (1H, bs), 8.21 (1H, bs), 7.83 (1H, d, J = 2, 2 Hz), 7.66 (1H, d, J = 9.0 Hz), 7.44 (1H, dd, J = 9.0, 2.2 Hz), 7.39 (1H, s), 7 28 (1H, d, J = 8.1Hz), 7.25 (1H, dd, 7.5, 7.2Hz), 7.01 (1H, d, J = 8.1Hz), 6 92 (1H, dd, J = 7.5, 7.2 Hz), 4.51 (2H, bs), 3.82 (3H, s).

FABMS (MH &lt; + &gt;): 414/416.

Anal. Calcd for C ₂₀ H ₁₆ N ₃ O ₃ ClS: C, 58.04; H, 3.90; N, 10.15; S, 7.75; Cl, 8.57. Found: C, 57.97; H, 3.85; N, 10.11; S, 7.85;

Cl, 8.63.

* Example C (32) [4-Amino-2- (1H-benzoimidazol-6-ylamino) -thiazol-5-yl] - (2-methoxyphenyl) -methanone

The title compound was prepared in a manner similar to that described in Example C (1). 6-Isothiocyanato-1H-benzoimidazole (see Boev et al., Pharm. Chem. J., vol.

(1990), pages 812-822) and 2-bromo-2'-methoxyacetophenone gave 72% yield of amorphous yellow powder, m.p. 180-185 ° C (dec.).

¹ H NMR (DMSO-d ₆ ): δ 12.40, (1H, bs), 10.61 (1H, bs), 8.16 (1H, s), 7.94 (2H, bs), 7, 83 (1 H, bs), 7.53 (1 H, d, J = 8.4 Hz), 7.36 (1 H, ddd, J = 8.4, 7.6, 1.6 Hz), 7.24 7.16 (2H, m), 7.05 (1H, d, J = 8.1Hz), 6.95 (1H, dd, J = 7.6, 7.2Hz), 3.74 ( 3H, s).

FABMS (MH &lt; + &gt;): 366.

Anal. calculated for C ₁₈ H ₁₅ N ₅ O ₂ S. 0.5 H ₂ O: C, 57.74; H, 4.31; N, r * yyyy r, - yyyy yy f 50

18.71; S, 8.56. Found: C, 57.78; H, 4.29; N, 18.64; S, 8.53. * Example C (33)

4- (4-Amino-5- (2,4-dimethoxy-benzoyl) -thiazol-2-ylamino) -benzenesulfonamide

The title compound was prepared essentially as described in Example C (1). 4-Isothiocyanate-benzenesulfonamide and 2-bromo-2 ', 4'-dimethoxyacetophenone gave a 75% yellow powder yield, m.p. Mp 249-250 ° C.

NMR (DMSO-d ₅ ): δ 10.93 (1H, bs), 7.93 (2H, bs), 7.75 (4H, bs), 7.25 (2H, bs), -7.21 ( 1H, d, J = 8.1 Hz), 6.61 (1H, d, J = 1.9 Hz), 6.55 (1H, dd, J% 8.1 Hz). 3.79 (3H, s); 3.76 (3H, s).

FABMS (MH &lt; + &gt;): 435.

Anal. calcd for C ₁₉ H ₁₃ N ₄ O ₅ S _2: C, 49.76; H, 4.18; N, 12.89; S, 14.86.

* Example C (34)

Ethyl 4- / 4-Amino-2- (4-sulfamoyl-phenylamino) -thiazole-5-carbonyl / benzoate

The title compound was prepared essentially as described in Example C (1). 4-Isothiocyanate-benzenesulfonamide and ethyl 4-bromoacetyl benzoate gave a 95% yield of a yellow powder, m.p. 225-227 [deg.] C.

¹ H NMR (DMSO-d ₆ ): δ 11.16 (1H, s), 8.32 (2H, bs), 8.04 (2H, d, J 8.4 Hz), 7.80 (2H, d, J = 8.4 Hz), 7.78 (4H, bs), 7.26 (2H, bs),

4.33 (2Η, q, J = 7.2 Hz), 1.33 (3H, t, J = 7.2 Hz).

FABMS (MH &lt; + &gt;): 447.

Anal. calculated for C ₁₉ H ₁₈ N ₄ O ₅ S ₂ . 0.4 H ₂ O: C, 50.30; H, 4.18; N, 12.38; S, 14.13. Found: C, 50.11; H, 3.97; N, 12.26; S, 14.14. *. Example C (35)

4- (4-Amino-5- (2,4-dimethyl-benzoyl) -thiazol-2-ylamino) -benzenesulfonamide

The title compound was prepared essentially as described in Example C (1). 4-Isothiocyanatobenzenesulfonamide and 2-bromo-2 ', 4'-dimethylacetophenone yielded 75% yield

yellow powder, m.p. Mp 242-244 ° C. ¹ H NMR (DMSO-d ₆ ): δ 10.97 (OH, bs), 8.00 (2H, bs) 7.76 (2 H, d, J) 9.7 Hz), 7.72 (2H, d, J = = 9.7 Hz), 7.24 (2H, bs) 7.22 (1 H, d, J) = 7.5 Hz), 7.07 (1H, s), 7.03 (OH, d, J = 7.5 Hz) 2.29 (3H, s).

2.23 (3 H, s).

FABMS (MH &lt; + &gt;): 403.

Anal. calcd for C ₁₈ H ₁₈ N ₄ O ₃ S _2: C, 53.71; H, 4.51; N, 13.92; S, 15.92. Found: C, 53.47; H, 4.54; N, 13.69; S, 15.83.

* Example C (36) [4-Amino-2- [4- (2-chloro-5-trifluoromethyl-pyridin-2-yl-sulfanyl) -phenylamino] -thiazol-5-yl] - (2,6-dichloro- 4-trifluoromethylphenyl) methanone nh ₂ n &lt; 52

The title compound was prepared essentially as described in Example C (1). 2- [4- (2-chloro-5-trifluoromethylpyridin-2-ylsulfanyl) phenyl] isothiocyanate; and

2-bromo-2 ', 6'-dichloro-4'-trifluoromethyl-acetophenone gave an orange solid in 52% yield, m.p. 130-132 [deg.] C.

Ή NMR (DMSO-d _s): δ 8.65 (IH, bs), 8.38 (IH, bs), 8.06 (2H, d, J = 8.0 Hz), 7.80 (IH, t, J = 7.0 Hz), 7.74-7.64 (4H, m), 7.54 (2H, d, J = 8.0 Hz).

IR (KBr): 3272, 3048, 1596, 1531, 1431, 1320 cm &^lt; -1 & gt ^; .

Anal. Calcd. for C ₂₂ H ₁₃ ClF ₃ N ₅ O ₃ S ₂ : C, 47.87; H, 2.37; N, 12.69; S, 11.62; Cl, 6.42. Found: C, 47.79; H, 2.44; N, 12.54; S, 11.70; Cl, 6.52.

* Example C (37) [4-Amino-2- (1H-benzoimidazol-6-ylamino) -thiazol-5-yl] - (2,6-dichloro-4-trifluoromethyl-phenyl) -methanone

The title compound was prepared in an analogous manner to that described in Example C (1). 6-Isothiocyanato-1H-benzoimidazole (see Boev et al., Pharm. Chem. J. (English translation)., Vol. 24 (1990), pp. 812-822) and 2-bromo-2 ', 6 1'-dichloro-4'-trifluoromethyl-acetophenone gave a yellow solid in 56% yield, which decomposed above 180 ° C.

¹ H NMR (DMSO-d 6): δ 12.45 (1H, bd, J = 16.0 Hz), 11.10-10.80 (1H, s), 8.00 (2H, s), 7, 70-7.45 (2H, m), 7.20 (1H, d, J = 8.0 Hz).

IR (KBr): 3191, 2974, 1619, 1559, 1467, 1309 cm &^lt; -1 & gt ^; .

FABMS (MH &lt; + &gt;): 472.

Anal. calcd for C ₁₀ H _ig Cl ₂ F ₃ N ₅ OS. 0.6 HOAc. 0.1 CH ₂ Cl ₂ . H ₂ O: C, e *

Γ. 53

45.58; Η, 2.95; Ν, 12.18; S, 5.69; Cl, 13.92. Found: C, 45.70; Η, 3.05; Ν, 12.45; Cl, 13.87.

* Example C (38)

4- (4-Amino-5- (2,6-dichloro-4-trifluoromethyl-benzoyl) -thiazol-2-ylamino) -benzenesulfonamide

The title compound was prepared essentially as described in Example C (1). 4-Isothiocyanatobenzenesulfonamide and 2-bromo-2 ', 6'-dichloro-4'- (trifluoromethyl) acetophenone gave, after recrystallization from EtOH / H ₂ O and drying over a benzene azeotrope, a yellow solid in 46% yield, mp 294- 296 ° C.

¹ H NMR (DMSO-d ₆ ): δ 8.10 (1H, s), 8.05 (2H, sj, 7.77 (4H, dd, J = 9.0, 14.0 Hz)).

HRFABMS (MH +): Calculated for C ₁₇ H ₁₂ Cl ₂ F ₃ N ₄ O ₃ S ₂ (MH +): 510.9680. Found: 510.9697.

Anal. calculated for C ₁₇ H ₁₂ Cl ₂ F ₃ N ₄ O ₃ S ₂ . 0.1 H ₂ 0. C _and H _5: C, 40.28; H, 2.51; N, 10.30; S, 11.97; Cl, 13.51. Found: C, 40.58; H, 2.28; N, 10.75; S, 12.31; Cl, 13.61.

* Example C (39)

Phenyl 4- / 4-Amino-2- (4-sulfamoylphenylamino) -thiazole-5-carbonyl / benzoate

The title compound was prepared in an analogous manner to that described in Example C (1). 4-Isothiocyanatobenzenesulfonamide and 4- (bromoacetyl) phenyl benzoate gave a yellow solid in 77% yield, m.p. above 300 ° C.

¹ H NMR (DMSO-d ₆ ): δ 11.13 (1H, s), 8.26 (2H, bs), 8.15 (2H, dd, J 7.2, 1.6 Hz), 7, 83-7.73 (7H, m), 7.66-7.59 (2H, m), 7.41 (2H, d, J = 6.9 Hz), 7.27 (2H, bs).

HRFABMS (MH +): Calculated: 95.0797. Found: 495.0812.

Anal. calculated for C ₂₃ H ₁₈ N ₄ O ₅ S ₂ . 0.2 H ₂ O: C, 55.45; H, 3.72; N, 11.25; S, 12.87. Found: C, 55.34; H, 3.592; N, 11.01; WITH,

12.83.

* Example 40:

(4-Amino-2- (4-methoxy-phenylamino) -thiazol-5-yl) - (4-methyl-1H-imidazol-5-yl) -methanone

5-Bromoacetyl-4-methyl-1H-imidazole having the structural formula: _u was prepared first, following

procedure. Bromine (0.40 mL, 7.77 mmol) was added dropwise to a solution of 5-acetyl-4-methyl-1H-imidazole (964 mg; 7.77 mmol; LaMattina et al., J. Org. Chem. , Vol 48 (1983), pp 897-898) in HOAc (20 mL). After two days, HOAc was removed in vacuo and the residue was partitioned with CH ₂ Cl ₂ and saturated NaHCO _{3. The} organic layer was washed with brine, dried over Na ₂ SO ₄ and evaporated to give a light brown solid, 625 mg (40% yield) which was used without further purification.

¹ H NMR (DMSO-d 6): δ 12.65 (1H, bs), 7.67 (1H, s), 4.62 (2H, s), 2.44 (3H, s).

The title compound was prepared in a manner analogous to that used in Example C (1). 4-Methoxyphenyl isothiocyanate and 5-bromoacetyl-4-methyl-1H-imidazole gave a yellow powder in 57% yield, m.p. Mp 245-50 ° C.

NMR (DMSO-d _6): δ 12.28 (1H, bs), 10.21 (1H, s), 8.00 (2H, bs), 7.56 (1H, s), 7.49 (2H d, J = 9.0 Hz), 6.94 (2H, d, J = 9.0 Hz), 3.75 (3H, s), 2.50 (3H, s).

HRFABMS (M + Na +): Calcd: 325.0844. Found: 352.0840.

Anal. Calcd for C ₁₅ H ₁₅ N ₅ O ₂ S. 0.5 H ₂ O: C, 53.24; H, 4.77; N, 20.70; S, 9.48. Found: C, 53.43; H, 4.78; N, 20.54; S, 9.38.

* Example C (41) [4-Amino-2- (4-imidazol-1-yl-phenylamino) -thiazol-5-yl] - (2,4-dimethyl-phenyl) -methanone

1- (4-Isothiocyanate-phenyl) -1H-imidazole, which has the structural formula N-N-N- _NCS, was prepared first. Into solution

1- (4-aminophenyl) -1H-imidazole (1.00g, 6.30 mmol; Venuti et al., J. Med. Chem., Vol. 31 (1988), pp. 2136-2145) in acetone (10 ml) a solution of thiophosgene (580 microliters, 7.6 mmo) in acetone (15 ml) and a solution of 25% Na ₂ CO ₃ (15 ml) were simultaneously added at 0 ° C. The mixture was stirred at 0 ° C for 0.5 hours and allowed to warm to room temperature for 1.5 hours. The acetate was removed under reduced pressure and the residue was diluted with water. The cream colored precipitate was filtered off, washed with water and dried under high vacuum to give 1.20 g (95% crude yield) of a light solid which was used without further purification.

¹ H NMR (DMSO-d ₆ ): δ 8.33 (1H, s), 7.81 (1H, s), 7.76 (2H, d, J = 8.8 Hz), 7.61 (2H , d, J = 8.8 Hz), 7.12 (1H, s).

The title compound was prepared in a manner analogous to that used in Example C (1). 1- (4-Isothiocyanat-phenyl) -1H-imidazole and 2-bromo-2 ', 4'-dimethylacetophenone e ρ.

P-P-P gave a yellow powder in 14% yield, m.p. 180-180.5 ° C.

NMR (DMSO-d6): δ 10.80 (1H, bs), 8.10 (1H, s), 8.02 (1H, bs), 7.68 (2H, d, J = 7.5 Hz) , 7.58. (2H, d, J = 9.0Hz), 7.20 (1H, d, J = 7.8Hz), 7.10-7.00 (2H, m), 2.28 (3H, s) 2.24 (3H, s).

IR (KBr): 3393, 3119, 2925, 1612, 1566, 1524, 1425 cm &^lt; -1 & gt ^; .

FABMS (MH &lt; + &gt;): 390.

Anal. calculated for C ₂₁ H ₁₉ N ₅ OS. 0.2 H ₂ O: C, 64.17; H, 4.97; N, 17.82; S, 8.16. Found: C, 64.14; H, 4.98; N, 17.68; S, 8.21.

* Example C (42):

(4-Amino-2- (4-imidazol-1-yl-phenylamino) -thiazol-5-yl) - (3-methyl-

The title compound was prepared in a manner similar to that described in Example C (1).

1- (4-Isothiocyanat-phenyl) -ΙΗ-imidazole (from Example C (41)} a

2-bromoacetyl-3-methyl-thiophene (from Example C (19)) gave a yellow solid in 83% yield, m.p. higher than 300 ° C.

¹ H NMR (DMSO-d ₆ ): δ 10.98 (1H, s), 8.25 (1H, s), 8.18 (1H, bs),

7.77 (1H, s), 7.72 (2H, J = 6.5Hz), 7.65 (1H, s), 7.62 (2H, J = 4.7), 7.10 (1H) , s), 6.98 (1H, d, J = 5.0Hz), 2.28 (3H, s).

IR (KBr): 3402, 3278, 3103, 2982, 1609, 1523, 1422, 1306 cm &^lt; -1 & gt ^; .

Anal. calcd for C ₁₇ H ₁₆ N ₅ OS _2: C, 56 67; H, 3.96; N, 18.36; S, 16.81. Found: C, 56.38; H, 4.06; N, 18.13; S, 16.67.

* Example C (43) [4-Amino-2- (1H-benzimidazol-5-ylamino) -thiazol-5-yl] - (1-methyl-1H-pyrrol-2-yl) methanone

/

Η

Ο

The title compound was prepared in an analogous manner to that described in Example C (1). 6-Isothiocyanato-1H-benzoimidazole (see Boev et al., Pharm. Chem. J. (Eng. Trans.)., Vol. 24 (1990), pp. 812-822) and 2-chloroacetyl-N-methylpyrrole ( Croce et al., Synthesis (1990), pp. 212-213) gave a yellow solid in 42% yield, mp 284-285 ° C.

¹ H NMR (DMSO-d ₆ ): δ 12.43 (1H, bs), 10.65 (IH, bs), 8.18 (IH, s), 7.94 (3H, bs), 7.55 (IH, d, J = 8.7 Hz), 7.27 (IH, dd, J = 8.7,

1.9 Hz), 6.92 (1H, m), 6.62 (IH, dd, J = 3.7, 2.1 Hz), 6.04 (IH, dd, J = 4.1, 2) 1 Hz, 3.80 (3H, s).

HRFABMS (MH +): Calcd: 339.1028. Found: 339.1024.

Anal. calcd for C _g H ₁₄ N ₆ OS. 0.3 H ₂ O: C, 55.90; H, 4.28; N, 24.45; S, 9.33. Found: C, 56.08; H, 4.28; N, 24.46; S, 9.33.

* Example C (44):

1 'Z4- / 4 ^- Amino-5- (3-methyl-thiophene-2-carbonyl) -thiazol-2-ylamino / phenyl / ethanone

The title compound was prepared essentially as described in Example C (1). 4-Acetylphenyl-isothiocyanate and 2-bromoacetyl-3-methyl-thiophene (from Example C (19)) gave a yellow solid in 89% yield, m.p. Mp 171-172 ° C.

¹ H NMR (DMSO-d ₆ ): δ 11.14 (1H, s), 8.22 (2H, bs), 7.95 (2H, d, J = prr r)

9.0 Hz), 7.76 (2H, d, J = 9.0 Hz), 7.62 (1H, d, J = 5.0 Hz, 7.00 (1H, d, J = 5.0) Hz), 2.53 (3H, s), 2.39 (3H, s).

IR (KBr): 3618, 3354, 3254, 3178, 3072, 1651, 1599, 1524, 1403, 1355, 1318, 1275, 1170 cm &^lt; -1 & gt ^; .

FABMS (MH &lt; + &gt;): 357.

Anal. calculated for C ₁₇ H ₁₅ N ₃ O ₂ S ₂ . 0.5 H ₂ O: C, 55.72; H, 4.40; N, 11.47; S, 17.50. Found: C, 55.92; H, 4.44; N, 11.51; S, 17.44. * Example C (45) trans-3RS-Amino-4RS- [4- (4-amino-5- (3-methylthiophene-2-carbonyl) -thiazol-2-ylamino) benzoyl] -dihydrofuran-2-one

The title compound was prepared essentially as described in Example C (1). The product 4-isothiocyanate-benzoyl-DL-homoserine-lactone and 2-bromoacetyl-3-methyl-thiophene (from Example C (19)) was extracted in 10% i-PrOH / CHCl ₃ . Flash column chromatography with increasing gradient of 2-3-4-5-6% MeOH / CH ₂ Cl ₂ gave a yellow solid in 43% yield, mp 162-163 ° C.

¹ H NMR (DMSO-d ₆ ): δ 11.05 (1H, s), 8.88 (2H, d, J = 8.1 Hz), 8.32 (2H, bs), 7.85 (2H J = 9.0 Hz, 7.74 (2H, d, J = 9.0 Hz), 7.61 (1H, d, J = 5.0 Hz), 6.99 (1H, d, J = 5.0 Hz), 4.73 (1H, q, J = 9.3 Hz), 4.40 (1H, ddd, J = 10.8, 8.7, 2.0 Hz), 4, 26 (1H, ddd, J = 10.2, 8.7, 6.7).

IR (KBr): 3413, 3284, 3084, 1773, 1637, 1608, 1524, 1413, 1313, 1254, 1181 cm ^-1 .

FABMS (MH &lt; + &gt;): 443.

Anal. calculated for C ₂ oH ₁₈ N ₄ 0 ₄ S _2nd 0.4 H ₂ O: C, 53.41; H, 4.21; N,

C r

12.46; S, 14.26. Found: C, 53.56; H, 4.28; N, 12.30; S, 14.43. * Example C (46):

Ethyl 3RS- [4-Amino-5- (3-methyl-thiophene-2-carbonyl) -thiazol-2-ylamino] -butyrate

The title compound was prepared essentially as described in Example C (1). The product ethyl dl-3-isothiocyanate-butyrate and 2-bromoacetyl-3-methyl-thiophene (from Example C (19)) was extracted with 10% i-PrOH / CHCl ₃ . Flash column chromatography with 3% MeOH / CH ₂ Cl ₂ gave a yellow solid in 45% yield, mp 129-130 ° C.

¹ H NMR (DMSO-d ₅ ): δ 8.61 (1H, d, J = 7.8Hz), 8.08 (2H, bs), 7.53 (IH, d, J = 5.0Hz) 6.94 (1H, d, J = 5.0Hz, 4.05 (2H, q, J = 7.2Hz), 2.33 (3H, s), 1.22-1.12 ( 6H, m).

IR (KBr): 3307, 3213, 1976, 1737, 1618, 1586, 1526, 1423, 1349, 1215, 1183, 1091 cm &^lt; -1 & gt ^; .

FABMS (MH &lt; + &gt;): 353.

Anal. Calcd. for C ₁₅ H ₁₉ N ₃ O ₃ S ₂ : C, 50.97; H, 5.42; N, 11.89; S, 18.41. Found: C, 50.81; H, 5.39; N, 11.72; S, 17.97.

* Example C (47):

4- / 4-Amino-5- (4-methyl-thiazole-5-carbonyl) -thiazol-2-ylamino / benzenesulfonamide

5-Bromoacetyl-4-methyl-thiazole, which has a structural formula, was prepared as described by Sych et al., J. Gen. Chem. USSR, r

Sv p sv. 32 (1996), pp. 970-975. Bromine (0.75 mL, 7.77 mmol) was added dropwise to a solution of 1- (4-methyl-thiazol-5-yl) ethanone (2.05 mg, 14.5 mmol; Ganapathi et al., Proc. Indiana Acad Sci Sci Part A, Vol 22 (1945), pages 362-378) in HOAc (3 mL). The mixture was stirred at 85 ° C for 1.5 hours and became a yellow cake. HOAc (3 mL) was added, and allowed to cool for 1.5 hours. The HOAc was removed in vacuo and the residue was partitioned between CH ₂ Cl ₂ and saturated aqueous NaHCO ₃ . The organic layer was washed with brine, dried over Na ₂ SO ₄ and evaporated to give a black solid, yield 1.3 g (41%), which was used without further purification.

¹ H NMR (CDCl 3): δ 8.85 (1H, s), 4.28 (2H, s), 2.81 (3H, s).

The title compound was prepared essentially as described in Example C (1). 4-isothiocyanate-benzenesulfonamide and 5-bromoacetyl-4-methyl-thiazole gave a brown solid in 31% yield, m.p. 262-266 [deg.] C.

1 H NMR (DMSO-d 6): δ 11.18 (1H, s), 9.08 (1H, s), 8.30 (2H, bs),

7.78 (4H, bs), 7.72 (2H, bs), 2.55 (3H, s).

Anal. calcd for C ₁₆ H ₁₃ N ₃ O ₅ S _3: C, 45.52; H, 3.31; N, 17.11; S, 24.32. Found: C, 42.28; H, 3.33; N, 17.15; S, 24.52.

* Example C (48):

4- / 4-Amino-5- (3-methyl-thiophene-2-carbonyl) -thiazol-2-ylamino / benzenesulfonamide

The title compound was prepared essentially as described in Example C (1). 4-Isothiocyanate-benzenesulfonamide and 2-bromoacetyl-3-methyl-thiophene (from Example C (19)) gave a yellow solid in 69% yield, m.p. 284.5-286.0 ° C.

Γ Γ <· << · «·; γ r <rr · 'r λ ^"r rr - r - ^{Χ Η} NMR (DMSO-d _6): δ 11.11 (ΙΗ, s), 8.20 (2Η, bs), 7.80 (2Η d J = 10.7 Hz, 7.76 (2H, d, J = 10.7 Hz), 7.61 (1H, d, J = 5.0 Hz), 7.26 (2H, s), 6.90 (1H, d, J = 5.0Hz); 2.38 (3H, s).

Anal. Calcd. for C ₁₅ H ₁₄ N ₄ O ₃ S ₃ : C, 45.67; H, 3.58; N, 14.20; S, 24.39. Found: C, 45.52; H, 3.58; N, 14.04; S, 24.36.

* Example C (49):

4- (4-Amino-5- (3-methyl-benzo) b / thiophene-2-carbonyl) -thiazol-2-ylamino / benzenesulfonamide

The title compound was prepared essentially as described in Example C (1). 4-Isothiocyanate-benzenesulfonamide and 2- (2-bromoacetyl) -3-methyl-benzo [b] thiophene gave a yellow solid powder in 73% yield, m.p. 274.0-275.5 ° C.

Δ NMR (DMSO-d 6): δ 11.17 (1H, bs), 8.33 (2H, bs), 8.04-7, 97 (1H, m, 7, 90-7.84 (1H, m), 7.78 (4H, bs), 7.51-7.44 (2H, m), 7.27 (2H, s), 2.52 (3H, s).

Anal. calcd for C ₁₉ H ₁₆ N ₄ O ₃ S _3: C, 51.33; H, 3.63; N, 12.60; S, 21.64. Found: C, 51.19; H, 3.67; N, 12.31; S, 21.37.

* Example C (50):

4- / 4-Amino-5- (2,5-dimethyl-thiophene-3-carbonyl) -thiazol-2-ylamino-benzenesulfonamide

3-Bromoacetyl-2,5-dimethyl-thiophene having the structural formula

Br ^- , was prepared in a manner analogous to step ₃

The process for the preparation of 2-bromo-2'-iodoacetophenone from Example C (12). 3-Acetyl-2,5-dimethylthiophene (6.83 g, 44.3 mmol) yielded 10.1 g (98% yield) of a yellow oil which was used without further purification.

¹ H NMR (CDCl 3): δ 7.22 (1H, s), 4.64 (2H, s), 2.58 (3H, s), 2.36 (3H, s).

The title compound was prepared essentially as described in Example C (1). 4-Isothiocyanate-benzenesulfonamide and 3-bromoacetyl-2,5-dimethyl-thiophene gave a yellow powder in

69% yield, m.p. Mp 163-265 ° C. ^X H NMR (DMSO-d: δ 11.02 (1 H, s), 8.05 (2H, bs), 7.76 (4H, s); 7.25 (2H, s), 6.87 (1H, s), 2.43 (3H, S), 2.38 (3H, s). Anal. calculated for Cíí ^ is is ^ Ojj: C, 47, 04; H, 3.95; N, 13.71; WITH, 23.55. Found: C, 47.01; H, 3.92 ; N, 13.62; S, 23.47.

* Example C (51):

4- / 4-Amino-5- (2-oxo-1,2,3,4-tetrahydroquinoline-6-carbonyl) -thiazol-2-ylamino / benzenesulfonamide

The title compound was prepared essentially as described in Example C (1). 4-Isothiocyanat-benzenesulfonamide and 6- (bromoacetyl) -2-oxo-1,2,3,4-tetrahydroquinoline gave a gray-yellow solid in 48% yield, m.p. 300-305 ° C (d).

¹ H NMR (DMSO-d ₆ ): δ 11.08 (1H, s), 10.32 (1H, s), 8.17 (2H, bs), 7.82-7.70 (4H, m) 7.58-7.45 (3H, m), 7.27 (1H, s), 6.90 (1H, d, J = 8.1 Hz), 2.93 (4H, t, J = 7) 7 Hz).

IR (KBr): 3266, 3193, 3069, 1679, 1597, 1525, 1434, 1365, 1317, 1153 cm &^lt; -1 & gt ^; .

HRFABMS: Calcd. For C ₁₉ H ₁₃ N ₅ O ₄ S ₂ (MH +): 444.0800. Found:

»! e e e r ρ

444.0816.

Anal. calculated for C ₁₉ H ₁₇ N ₅ O ₄ S ₂ . 0.6 MeOH: C, 50.88; H, 4.23; N, 15.13; S, 13.86. Found: C, 51.02; H, 4.00; N, 15.00; S, 13.60.

* Example C (52):

4-Amino-2- (1H-benzoimidazol-6-ylamino) thiazol-5-yl- (2,6-dichloro-phenyl) -methanone

ΛΛ2-Bromo-2,6-dichloroacetophenone having the structural formula:

Cl was prepared as follows: To 2 ', 6'-dichloroacetophenone (1.0 g, 5.30 mmol) in HOAc (5 mL) was added dropwise bromine (272 microliter,

5.30 mmol). The mixture was heated to 90 ° C for 1 hour, then diluted with ice water and partitioned between ether and saturated aqueous NaHCO ₃ . The organic layer was washed with brine, dried over MgSO _4, concentrated and azeotroped twice with heptane to yield 1.41 g (100% yield) of a pale yellow oil which corresponds to ^X H NMR and IR previously described (see Mlotkowska et al., Pol .

J. Chem., Vol. 55 (1981), pp. 631-642) and was used without further purification.

¹ H NMR (CDCl ₃ ): δ 7.39-7.33 (3H, m), 4.23 (2H, s).

The title compound was prepared in an analogous manner to that described in Example C (1). 6-Isothiocyanato-1H-benzoimidazole (see Boev et al., Pharm. Chem. J. (English trans.)., Vol. 24 (1990), pp. 812-822) and 2-bromo-2 ', 6 Dichloroacetophenone (from Example C (52)) gave a yellow solid in 47% yield, mp 203-208 ° C.

¹ H NMR (DMSO-d ₆ ): δ 12.47 (1H, d, J = 17.7 Hz), 10.83 (1H, d, J =

16.5 Hz), 8.22-7.80 (3H, m), 8.18 (1H, s), 7.76-7.36 (5H, m);

Γ <- r '* rrrp P PP ^r

Γ - * cr. ' rf · - rr - Γ fr. Γ · P rp ^A _ ^r

7.19 (1H, d, J = 8.4 Hz).

Anal. calcd for C ₁₇ H ₁₂ N ₅ OSC1 _2: C, 50.51; H, 2.74; N, 17.32; S, 7.93; Cl, 17.54. Found: C, 50.32; H, 2.78; N, 17.11; S, 7.91; Cl, 17.75.

* Example C (53):

[4-Amino-2- [4- (1H-imidazol-2-yl) -phenylamino] -thiazol-5-yl] - (2,6-dichloro-4-trifluoromethyl-phenyl) -methanone

2- (4-Nitrophenyl) -1H-imidazole having the structural formula:

No ₂ was first prepared as follows: To a solution of 2-phenylimidazole (5.00 g, 34.7 mmol) in conc. H ₂ SO ₄ (20 mL) was added conc. HNO ₃ (2.2 mL, 35 mmol) in conc. H ₂ SO ₄ (5 mL). The resulting brown mixture was stirred at 0 ° C for 2 hours and cooled with crushed ice. A pale white precipitate formed and was filtered. The filtrate was adjusted to pH 9 with 2N NaOH. A yellow precipitate formed, which was filtered off, washed with H ₂ O and recrystallized from boiling MeOH to give 3.0 g (46% yield) of a yellow solid. This crude product was used without any further purification.

^{Σ Η} NMR (MeOH-d _4): δ 8.34 (2H, d, J = 9.0 Hz), 8.08 (2H, d, J = 9.0 Hz), 7.26 (2H, s ).

t / v ^{NH 2}

4- (1H-Imidazol-2-yl) aniline having the structural formula: h was further prepared as follows. Into suspension

2- (4-nitro-phenyl) -1H-imidazole (1.5 g, 7.93 mmol) in absolute ethanol (30 mL) was added 10% Pd-C (250 mg). The resulting mixture was stirred under H ₂ for 5 hours. The mixture was filtered through a pad of Celite. The filtrate was concentrated under reduced pressure to give 1.20 g (95% crude oil yield) of a red resin which was used without further purification.

2- (4-Isothiocyanato-efnyl) -1H-imidazole having the structural formula:

was prepared in a manner analogous to the preparation of 1- (4-isothiocyanato-phenyl) -1H-imidazole from Example C (41). 4- (lH-imidazol-2-yl) -aniline gave a pale brown solid which was recrystallized from CHC1 ₃ in 85% yield and was used without further purification.

¹ H NMR (MeOH-dJ: δ 7.88 (4H, bd, J = 7.8 Hz), 7.58 (2H, s).

The title compound was prepared essentially as described in Example C (1). 4-Isothiocyanat-phenyl) 1H-imidazole and 2-bromo-2 ', 6'-dichloro-4'-trifluoromethyl-acetophenone gave, after purification by preparative thin layer chromatography with MeOH: CHCl ₃ (8:92) as eluent, a yellow solid in 21% yield, mp 195-197 ° C (d).

¹ H NMR (DMSO-d ₆ ): δ 11.10 (1H, s), 8.18 (1H, s), 8.02 (2H, s), 7.88 (2H, d, J = 8, 7 Hz), 7.62 (2H, d, J = 8.1 Hz), 7.12 (2H, bs).

IR (KBr): 3400, 2929, 1610, 1527, 1426, 1310 ^cm-first

HRFABMS: Calcd for C 24 H 31 Cl 2 AOS (MH +): 498.0170. Found: 498.0183.

Anal. calculated for C ₂₀ H ₁₂ Cl ₂ F ₃ N ₅ OS. H ₂ O: C, 46.52; H, 2.73; N,

13.56; Cl, 13.73; S, 6.21. Found: C, 46.45; H, 2.78; N

13.40; Cl, 13.73; S, 6.11.

* Example C (54):

(4-Amino-2- (4-morpholin-4-yl-phenylamino) -thiazol-5-yl) - (2,4-dimethyl-phenyl) -methanone

O

4- (4-Isothiocyanato-phenyl) morpholine having the structure:

, was prepared subsequently. To 4-morpholinaniline (2.0 g, 11.2 mol) and triethylamine (5.01 mL, 35.9 mmol) in THF (200 mL) at 0 ° C was added dropwise thiophosgene (1.03 mL, 13 mL). , 5 mmol). The mixture was stirred at ambient temperature overnight, and then partitioned between ether and water. The ether layer was washed with water and brine, dried over MgSO ₄ and concentrated to give 2.46 g (99%) of a dark brown solid.

¹ H NMR (CDCl ₃ ): δ 7.15 (2H, d, J = 9.3 Hz), 6.87 (2H, d, J = 9.3 Hz), 3.80 (4H, t, J = 5.0 Hz), 3.19 (4H, t, J = 5.0 Hz).

The title compound was prepared in a manner analogous to that used in Example C (1). 4- (4-Isothiocyanato-phenyl) morpholine and 2-bromo-2 ', 4'-dimethylacetophenone gave a yellow solid in 28% yield, m.p. 253-254.5 ° C.

1 H NMR (DMSO-d ₆ ): δ 10.44 (1H, s), 7.98 (2H, bs), 7.31 (2H, d, J = 9.0 Hz), 7.14 (1H, s, d, J = 7.8 Hz), 7.02 (1H, s), 6.99 (IH, d, J = 7.8 Hz), 6.90 (2H, d, J = 9.0 Hz) 3.70 (4H, t, J = 4.7Hz), 3.04 (4H, t, J = 4.7Hz), 2.26 (3H, s), 2.20 (3H, s) .

Analyt. Calcd for C ₂₂ H ₂₄ N ₄ O ₂ S: C, 64.68; H, 5.92; N, 13.71; S, 7.85. Found: C, 64.49; H, 5.97; N, 13.64; S, 7.93.

* Example C (55):

[4-Amino-2- (4-morpholin-4-yl-phenylamino) -thiazol-5-yl] - (2,6-dichloro-phenyl) -methanone

The title compound was essentially prepared as described in the Example. C (1). 4- (4-Isothiocyanat-phenyl) morpholine (from Example C (54)) and 2-bromo-2 ', 6'-dichloroacetophenone (from r *

of Example C (52)) gave a yellow solid in 9% yield, m.p. Mp 245-275 ° C (d).

NMR (DMSO-d _6): δ 10.58 (lH, .s), 8.02 (2H, bs), 7.52 (2H, d, J = 7.3Hz), 7.41 (1H, m), 7.30 (2H, d, J = 9.0 Hz), 6.92 (2H, d, J = 9.0 Hz), 3.72 (4H, dd, J = 5.0, 4) 2 Hz), 3.06 (4H, dd, J = 5.0,

4.2 Hz).

Anal. Calcd for C ₂₀ H ₁₈ N ₄ O ₂ SC 1: C, 53.46; H, 4.04; N, 12.47; S, 7.14; Cl, 15.78. Found: C, 53.39; H, 4.04; N, 12.47; S, 7.21; Cl, 15.71.

* Example C (56):

Ethyl 4- / 4-amino-5- (2,6-dichlorobenzoyl) -thiazol-2-ylamino / benzoate

The title compound was prepared in an analogous manner to that described in Example C (1). 4-Ethoxycarbonylphenyl isothiocyanate and 2-bromo-2 ', 6'-dichloroacetophenone (from Example C (52)) gave an amorphous yellow solid in 48% yield.

1 H NMR (DMSO-d 6): δ 11.13 (1H, s), 8.15 (2H, bs), 7.92 (2H, d, J = 8.7 Hz), 7.70 (2H, d, J = 8.7 Hz), 7.58-7.40 (3H, m), 4.27 (2H, q, J% 7.0 Hz), 1.29 (3H, t, J = 7 , 0 Hz).

Anal. calculated for C _1? H ₅ N ₃ O ₃ SC1 _2: C, 52.30; H, 3.47; N, 9.63; S, 7.35; Cl, 16.25. Found: C, 52.50; H, 3.42; N, 9.63; S, 7.44; Cl, 16.26.

* Example C (57):

(4-Amino-2- (1H-benzoimidazol-6-ylamino) thiazol-5-yl) - (2,4,6-trimethylphenyl) methanone

r ρ 'e e

2-Bromo-2 ', 4', 6'-trimethyl-acetophenone having the structural formula h ₃

HaCl ₃ was prepared in a manner analogous to the method for preparing 2-bromo-2'-iodoacetophenone, see Example C (12). 2,4,6-Trimethylacetophenone (1.50 g, 9.25 mmol) gave 2.26 g (100%) of a clear oil, which was used without further purification.

1 H NMR (CDCl ₃ ): δ 6.87 (2H, s), 4.27 (2H, s), 2.22 (9H, s).

The title compound was prepared in an analogous manner to that described in Example C (1). 6-Isothiocyanato-1H-benzoimidazole (see Boev et al., Pharm. Chem. J. (English trans.)., Vol. 24 (1990), pp. 812-822) and 2-bromo-2 ', 4 6'-trimethyl-acetophenone gave a yellow powder in 26% yield, which decomposed above 185 ° C.

¹ H NMR (DMSO-d ₆ ): δ 12.42 (1H, bs), 10.66 (1H, bs), 8.17 (1H, S), 7.96 (2H, bs), 7.75 (1H, bs), 7.44 (1H, bs), 7.16 (1H, d, J = 8.7), 6.82 (2H, s), 2.21 (3H, s), 2, 11 (6 H, s).

HRFABMS (MH +): Calcd .: 378.1389. Found: 378.1381.

Anal. calculated for C ₂₀ H ₁₉ N ₅ OS. 0.3 H ₂ O: C, 62.74; H, 5.16; N, 18.29; S, 8.37. Found: C, 62.96; H, 5.14; N, 18.24; S, 8.35.

* Example C (58):

(4-Amino-2- (1H-benzoimidazol-6-ylamino) thiazol-5-yl) - (2,3,6-trimethylphenyl) methanone

formula:

2-Bromo-2 ', 3', 6'-trimethyl-acetophenone, which has the structural ^Br_ ^^ ^ H3 _CH3 Prepared analogously to

2-bromo-2'-iodoacetophenone in Example C (12). 2 ', 3', 6'-trimethylacetophenone (1.50 g, 9.25 mmol) gave 2.10 g (93%) of a clear oil, which was used without further purification.

The title compound was prepared in an analogous manner to that described in Example C (1). 6-Isothiocyanato-1H-benzoimidazole (see Boev et al., Pharm. Chem. J. (English trans.)., Vol. 24 (1990), pp. 812-822) and 2-bromo-2 ', 3 6'-trimethyl-acetophenone gave a yellow powder in 70% yield, which decomposed above 196 ° C.

Ή NMR (DMSO-d ₆ ): δ 12.41 (1H, bs), 10.65 (IH, bs), 8.17 (1H, s), 7.96 (2H, bs), 7.70 (1H, bs) 1H, bs), 7.52 (IH, bs), 7.17 (1H, dd, J = 8.4, 1.9 Hz), 6.82 (2H, s), 2.11 (9H, s) ).

Anal. calculated for C ₂₀ H ₁₉ N ₅ OS: C, 63.64; H, 5.07; N, 18.55; S, 8.50. Found: C, 63.40; H, 5.17; N, 18.37; S, 8.36.

* Example C (59):

(4-Amino-2- (4-sulfamoyl-phenylamino) -thiazol-5-yl) - (3,5-dimethyl-pyridin-4-yl) -methanone

First, 4- (bromoacetyl) -3,5-dimethylpyridine hydrobromide, which has the structural formula: Her Bii ^^ * ³ as follows. h ₃ ct ^ - <n ' ^HBr

4-Acetyl-3,5-dimethylpyridine. (500 mg, 3.36 mmol; Kutney et al., Can.J.Chem., Vol. 41 (1963), pp. 695-702) was dissolved in 30% HBr in acetic acid (1 mL), warmed to 70 ° C and treated with a mixture of bromine (0.17 mL, 3.36 mmol) in 30% HBr in acetic acid (0.5 mL). After 2 hours, the mixture was allowed to cool to ambient temperature and ether (8 mL) was added. The resulting precipitate was filtered, washed with ether (2x), and dried to give 1.03 g (100%) of a violet solid, m.p. 222-225 ° C, which was used without further purification.

⁷⁰

The title compound was prepared essentially as described in Example C (1). 4-Isothiocyanato-benzenesulfonamide and 4- (bromoacetyl) -3,5-dimethylpyridine hydrobromide gave a solid which was purified by column chromatography with 10% MeOH / CHCl ₃ and crystallized from MeOH to give 35 mg (51%) of an amorphous yellow solids.

¹ H NMR (DMSO-d ₅ ): δ 11.09 (1H, s), 8.32 (1H, s), 8.18 (2H, bs),

7.74 (4H, dd, J = 11.5, 9.3 Hz), 7.27 (2H, s), 2.15 (6H, s).

IR (KBr): 3378, 3342, 3260, 3160, 1625, 1594, 1560, 1518, 1443, 1342, 1160 cm · ^1st

HRFABMS: Calcd. for C ₁₇ H ₁₈ N ₅ O ₃ S ₂ (MH +): 404.0851. Finding. : 404.0840

Anal. calculated for C ₁₇ H ₁₇ N ₅ O ₃ S ₂ . 0.4 H ₂ O. 0.3 MeOH: C, 49.44; H, 4.56; N, 16.66; S, 15.26. Found: C, 49.13; H, 4.31; N, 16.61;

S, 15.10.

* Example C (60):

[4-Amino-2- (1H-benzoimidazol-6-ylamino) -thiazol-5-yl] - (2,6-dimethyl-phenyl) -methanone

2 ', 6'-Dimethylacetophenone having the structural formula:

was prepared by the procedure for o-nitro-acetophenone (Reynolds et al., Org.Syn.Coli., Vol. IV (1963), pp. 708-710). 2,6-Dimethylbenzoic acid (3.00 g, 20.0 mmol) yielded 2.56 g (86% yield) of a yellow oil which was used without further purification.

¹ H NMR (CDCl ₃ ): δ 7.16 (1H, t, J = 7.2 Hz), 7.02 (2H, d, J = 7.2 Hz), 2.48 (3H, s), 2.25 (6 H, s).

^Βγ '^> Ύ

2-Bromo-2 ', 6'-dimethyl-acetophenone, structural formula:

was prepared in a manner analogous to 2-bromo-2'-acetophenone, see Example C (12). 2 ', 6'-dimethylacetophenone (1.50 g, 10.1 mmol) yielded 2.04 g (89% yield) of a clear oil, which was used without further purification.

¹ H NMR (CDCl ₃ ): δ 7.21 (1H, t, J = 7.2 Hz), 7.05 (2H, t, J = 7.2

Hz), 4.29 (2H, s), 2.26 (6H, s).

The title compound was prepared in an analogous manner to that described in Example C (1). 6-Isothiocyanato-1H-benzoimidazole (see Boev et al., Pharm. Chem. J. (English translation)., Vol. 24 (1990), pp. 812-822) and 2-bromo-2 ', 6 1-Dimethylacetophenone gave a yellow solid in 71% yield, which decomposed above 185 ° C.

* H NMR _(DMSO-D): δ 12.41 (1H, bs), 10.67 (1H, bs), 8.17 (1H, s), 7.99 (2H, s), 7.60 (1H, s), 7.52 (1H, s), 7.17 (1H, dd, J = 8.7, 1.9 Hz), 7.12 (1H, d, 7.1 Hz), 7 02 (1H, d, J = 7.5 Hz), 2.15 (6H, s).

HRFABMS (MH +): Calcd .: 364.1232. Found: 364.1227.

Anal. calculated for C ₁₉ H ₁₇ N ₅ OS. 0.3 CH ₃ OH: C, 62.14; H, 4.92; N, 18.77; S, 8.60. Found: C, 62.43; H, 5.15; N, 18.91; S, 8.60.

* Example C (61):

(4-Amino-2- (1H-benzoimidazol-6-ylamino) thiazol-5-yl) - (2-methyl-6-nitrophenyl) methanone

2'-methyl-6'-nitroacetophenone, the structural formula: ₂ n '^^ _J was prepared according to the procedure for o-nitroacetophenone (see Reynolds et al., Org.Syn.Coli., Vol. IV (1963 2-Methyl-6-nitrobenzoic acid (15.0 g, 82.8 mmol) gave 14.7 g (99% yield) of a yellow oil which was used without further purification.

1 H NMR (CDCl 3): δ 8.04 (1H, d, J = 8.4 Hz), 7.55 (1H, d, J = 7.5 Hz), 7.44 (1H, dd, J = 8.4, 7.5 Hz), 2.56 (3H, s), 2.35 (3H, s).

2-Bromo-2'-methyl-6'-nitroacetophenone having the structural formula: and ^AA, was prepared in a manner analogous to 5-bromoacetyl-4-methyl-lH-imidazole for Example C (40). Crude 2'methyl-6'-nitroacetophenone (1.56 g, 8.72 mL) gave a white solid, 2.17 g (97% yield), which was used without further purification.

¹ H NMR (CDCl 3): δ 8.11 (1H, d, J = 7.8Hz), 7.62 (1H, d, J = 7.8Hz), 7.52 (1H, d, t = 7.8 Hz), 4.33 (2H, s), 2.40 (3H, s).

The title compound was prepared in an analogous manner to that described in Example C (1). 6-Isothiocyanato-1H-benzoimidazole (see Boev et al., Pharm. Chem. J. (English trans.)., Vol. 24 (1990), pp. 812-822) and 2-bromo-2'-methyl -6'-nitroacetophenone gave a brown solid in 32% yield, mp Mp 198-201 ° C.

1 H NMR (DMSO-d 6): δ 12.40 (1H, bs), 10.78 (1H, bs), 8.17 (1H, d, J = 10.6 Hz), 8.00 (2H, bs) 7.92 (2H, d, J = 8.4 Hz), 7.68 (1H, d,

J = 7.5 Hz), 7.62-7.44 (2H, m), 7.19 (1H, d, J = 7.5 Hz), 2.30 (3H, s).

HRFABMS (MH +): Calcd .: 395.0926. Found: 395.0920.

Anal. calcd for C ₁₈ H ₁₄ N ₆ O ₃ S 0.5 H ₂ O: C, 53.59; H, 3.75; N,

20.83; S, 7.95. Found: C, 53.43; H, 3.67; N, 20.68; S, 7.81.

* Example C (62):

[4-Amino-2- (4-morpholin-4-yl-phenylamino) -thiazol-5-yl] - (2,6-dimethyl-phenyl) -methanone

«Γ Γ r« · r r ρ <r Γ

The title compound was prepared in a manner analogous to Example C (1). 4- (4-Isothiocyanato-phenyl) -morpholine (from Example C (54)) and 2-bromo-2 ', 6'-dimethylacetophenone (from Example G (60)) gave a brown solid in 23% yield, m.p. Mp 221-223 ° C.

¹ H NMR (DMSO-d ₆ ): δ 10.42 (1H, s), 7.95 (2H, bs), 7.30 (2H, d, J 9.0 Hz), 7.18-7, 10 (1H, m), 7.02 (2H, d, J = 7.5Hz), 6.91 (2H, d, J = 9.0Hz), 3.72 (4H, t, J = 4) 8 Hz), 3.05 (4H, t, J = 4.8 Hz), 2.16 (6H, s).

HRFABMS (M +): Calcd .: 408.1620. Found: 408.1607.

Anal. Calcd for C ₂₂ H ₂₄ N ₄ O ₂ S. 0.75 H ₂ O: C, 62.61; H, 6.09; N, 13.28; S, 7.60. Found: C, 62.64; H, 6.10; N, 13.05; S, 7.55.

* Example C (63):

[4-Amino-2- (1H-benzoimidazol-5-ylamino) -thiazol-5-yl] - (3,5-dichloro-pyridin-4-yl) -methanone

4-Bromoacetyl-3,5-dichloropyridine, having the structure:

was first prepared as follows: A mixture of 3,5-dichloropyridine-4-carboxylic acid (4.00 g, 20.9 mmol; Cale et al.,

J. Med. 32 (1989), pages 2178-2199), benzene (20 mL), DMF (0.4 mL) and thionyl chloride (3.80 mL, 52.0 mmol) was heated under reflux for 60 min, allowed to cool to room temperature. ambient, concentrated in vacuo, suspended in ether (20 mL), and carefully treated with a solution of trimethylsilyldiazomethane (25 mL of 2.0 M in hexane). After 72 hours, 48% HBr (18 mL) was carefully added dropwise over 20 min, with vigorous gas evolution. After 30 minutes, the mixture was carefully basified with NaHCO ₃ and extracted with ether. The ether layer was dried over Na ₂ SO ₄ and evaporated to give an orange oil which was purified by column chromatography eluting with 50% CH ₂ Cl ₂ / hex. to separate 2.50 g (51%)

Of 3,5-dichloropyridine-4-carbonyl chloride as a yellow oil, and yielding the desired product, 2.00 g (36%) of pale yellow crystals that darkened at ambient temperature and were used without further purification.

NMR (CDCl ₃ ): δ 8.58 (2H, s), 4.37 (2H, s).

Anal. calculated for C ₇ H ₄ BrCl ₂ NO. 0.02 C ₆ H ₁₄ : C, 31.60; H, 1.59; N, 5.18. Found: C, 31.92; H, 1.59; N, 5.24.

The title compound was prepared in an analogous manner to that described in Example C (1). 6-Isothiocyanato-1H-benzoimidazole (see Boev et al., Pharm. Chem. J. (English trans.)., Vol. 24 (1990), pp. 812-822) and

4- (bromoacetyl) -3,5-dichloropyridine gave the product, which was extracted in 10% MeOH / CHCl ₃ and column chromatography the same to give a yellow amorphous solid, 198 mg (55%). An analytical sample was precipitated from EtOH, mp 235-240 ° C (d).

1 H NMR (CD ₃ OD): δ 8.60 (2H, s), 8.18 (1H, s), 7.98 (IH, bs), 7.58 (IH, d, J = 9.0 Hz), 7.30 (1H, dd, J = 1.2, 8.7 Hz).

IR (KBr): 3183, 1608, 1544, 1461, 1427, 1355 cm &^lt; -1 & gt ^; .

HRFABMS: Calcd. for C ₁₆ H _n Cl ₂ N ₆ OS (MH +): 405.0092. Found: 405.0079.

Anal. calculated for C ₁₆ H ₁₀ Cl ₂ N ₆ OS. 1.1 H ₂ O: C, 45.51; H, 2.89; N, 19.77, Cl, 16.68, S, 7.54. Found: C, 45.49; H, 2.59; N, 19.64; Cl, 16.62; S, 7.43.

* Example C (64):

2S- [4-Amino-2- (1H-benzoimidazol-5-ylamino) -thiazole-5-carbonyl] -N-carbobenzyloxy-pyrrolidone

2S-Bromoacetyl-N-carbobenzyloxy-pyrrolidine having the structural formula

The MRO was prepared by the first following procedure. N-carbobenzyloxy-L-proline acid chloride (1.20 g, 4.80 mmol) was prepared according to Aoyama et al., Chem. 29 (1981), pp. 3249-3255, with oxalyl chloride and a catalytic amount of DMF. To a solution of the crude acid chloride in THF (5 mL) and MeCN (5 mL) at 0 ° C was carefully added dropwise a solution of trimethylsilyldiazomethane (5.0 mL in 2.0 M in hex.), And initially vigorous gas evolution was observed. The resulting red suspension was allowed to warm to room temperature and was stirred overnight. Then the brown mixture was allowed to cool to 0 ° C, carefully treated with a mixture of 47% HBr (4.1 mL) and ether (10 mL), followed by vigorous gas evolution. The mixture was allowed to warm to ambient temperature over 1 hour, then basified with saturated aqueous NaHCO ₃ (20 mL) and extracted with EtOAc (2 x 20 mL). The combined organic layers were dried over Na ₂ SO ₄ and evaporated to give a brown oil, 1.57 g (100%), which was used without further purification.

NMR (CDCl ₃ ): δ 7.44-7.24 (5H, m), 4.34 (1H, d, J = 15.6 Hz), 4.27 (1H, d, J = 15.6 Hz) ).

The title compound was prepared in an analogous manner to that described in Example C (1). 6-Isothiocyanato-1H-benzoimidazole (see Boev et al., Pharm. Chem. J., Vol. 24 (1990), pp. 812-822) and 2S-bromoacetyl-N-carbobenzyloxy- pyrrolidone gave a solid that was precipitated twice from iPrOH / hex to give a yellow amorphous solid, 154 mg (54%), mp 150-165 ° C.

^Ή NMR (DMSO-d 6): δ 12.40 (1H, d, J = 7.8 Hz), 10.68 (1H, d, J =

19.3 Hz), 8.20 (1H, d, J = 0.6 Hz), 8.10-7.70 (2H, m), 7.52 (1H, dd, J = 8.7, 34.8) Hz), 7.54-7.05 (5H, m), 5.17-4.80 (2H, m), 4.32 (1H, d, J = 4.9 Hz), 4.30-4 18 (1H, bm), 2.33-1.70 (2H, bm).

r r ''

IR (KBr): 3278, 1686, 1599, 1560, 1421, 1356, 1121 cm &^lt; -1 & gt ^; .

HRFABMS: Calcd. for C ₂₃ H ₂₃ N ₅ O ₃ S (MH +): 463.1552. Found: 463.1538.

Anal. calcd for C ₂₃ H ₂₂ N ₆ O ₃ S 0.1 H ₂ 0 .0,7 iPrOH: C, 59.53; H, 5.53; N, 16.60; S, 6.63. Found: C, 59.53; H, 5.53; N, 16.60; S, 6.22.

* Example C (65):

2S- [4-Amino-2- (4-sulfamoyl-phenylamino) -thiazole-5-carbonyl] -N-carbobenzyloxy-pyrrolidine nh ₂

The title compound was prepared essentially as described in Example C (1). 4-Isothiocyanate-benzenesulfonamide and 2S-bromoacetyl-N-carbobenzyloxy-pyrrolidine (see Example C (64)) gave a solid which was purified by column chromatography with 5% MeOH / CHCl ₃ to give a yellow amorphous solid, 140 mg ( 46%), mp 150-160 ° C (d).

¹ H NMR (DMSO-d ₆ ): δ 11.05 (1H, d, J = 10.0 Hz), 7.98 (2H, bd, J = 17.1 Hz), 7.79 (4H, dd) J = 12.1, 9.7 Hz), 7.41-7.11 (5H, m), 5.15-4.89 (2H, m), 4.37-4.21 (1H, bm) 3.51-3.40 (2H, bm), 2.35 2.13 (1H, bm), 1.93-1.75 (3H, bm).

IR (KBr): 3288, 1686, 1598, 1550, 1427, 1420, 1157 ^cm-first

HRFABMS: Calcd. for C ₂₂ H ₂₃ N ₅ O ₅ S ₂ Cs (M + Cs +): 634.0195. Found: 634.0215.

Anal. calculated for C ₂₂ H ₂₃ N ₅ O ₅ S ₂ . 0.3 H ₂ O • 0.1 CHCl ₃ : C, 51.15; H,

4.60; N, 13.50; S, 12.36. Found: C, 51.36; H, 4.63; N, 13.31;

S, 12.47.

Example C (66):

(4-Amino-2- (1H-benzoimidazol-6-ylamino) -thiazol-5-yl) - (2-bromo-6-methyl-phenyl) -methanone.

2'-Bromo-6'-methyl-acetophenone having the structural formula:

was prepared in a manner similar to o-nitro-acetophenone (see Reynolds et al., Org. Syn. Coli., Vol. IV (1963) pp. 708-710). From 2-methyl-6-bromobenzoic acid (3.10 g, 14.4 mmol), 2.45 g (80% yield) of a yellow oil was obtained which corresponded to the previously described material Ή NMR (Swenton et al., J. Org Chem., Vol. 58 (1993), pp. 3308-3316) and was used without further purification.

2,2'-Dibromo-6'-methyl-acetophenone having the structural formula:

was prepared in a manner analogous to 2-bromo-2'-iodoacetophenone, see Example C (12). Crude 2'-bromo-6'-methylacetophenone (1.00 g, 4.69 mmol) gave 1.48 g of a yellow oil, which was used without further purification.

¹ H NMR / CDCl 3: δ 7.44-7.37 (1H, m), 7.21-7.17 (2H, m), 4.24 (2H, s), 2.31 (3H, s) ).

The title compound was prepared in a manner analogous to that used in Example C (1). 6-Isothiocyanato-1H-benzoimidazole (see Boev et al., Pharm. Chem. J. (Eng. Trans.)., Vol. 24 (1990), pp. 812-822) and 2,2'-dibromo-6 ' methyl acetophenone gave a brown solid, in 32% yield, mp 208-210 ° C.

¹ H NMR (DMSO-d 6): δ 12.43 (1H, bs), 10.74 (1H, bs), 8.18 (1H, s), 8.02 (2H, s), 7.75 ( 1H, bs), 7.4 (lH, bs), 7.44 (1H, d, J = 7.5 ρ Γ ·· r rr r R _e rrfr yyyy · · Γ fro RFR

Hz), 7.28-7.14 (3H, m), 2.22 (2H, s).

ESIMS: (MH &lt; + &gt;): 428/430.

Anal. calcd for C ₁₈ H ₁₄ N ₅ OSBr. 1.0 H ₂ O: C, 48.44; H, 3.61; N, 15.69; S, 7.18, Br, 17.90. Found: C, 48.54; H, 3.69; N, 15.57; S, 7.11; Br, 17.88.

* Example C (67):

(4-Amino-2- (1H-benzoimidazol-6-ylamino) -thiazol-5-yl) - (3-methylbiphenyl-2-yl) methanone

1- (3-Methyl-biphenyl-2-yl) ethanone, structural formula:

was. prepared as follows. To 2'-bromo-6'-methyl-acetophenone (from Example C (66)); 760 mg, 3.58 mmol) and Pd (OAc) ₂ (114 mg) in DME (38 mL) at 0 ° C and under Ar were added excess phenylboronic acid (495 mg) and 2M aqueous Na ₂ CO ₃ (1 , 6 ml). The mixture was heated to 90 ° C for 3 hours, then diluted with water (50 mL) and extracted with ether (2 x 100 mL). The ether extracts were concentrated to the crude product, which was purified by column chromatography with a 2-5% ether / hexane step gradient to give 670 mg (89% yield) of a yellow oil, used without further purification.

¹ H NMR (CDCl ₃ ): δ 7.44-7.31 (5H, m), 7.25-7.19 (2H, m), 7.16-7.09 (1H, m), 2, 33 (3 H, s), 1.93 (3 H, s).

2- Bromoacetyl-3-methyl-biphenyl having the structural formula:

was prepared in a manner analogous to 2-bromo-2'-iodoacetophenone, see Example C (12)), Crude 1- (3-methyl-biphenyl-2-yl) ethanone (295 mg, 1.40 mmol) gave yellow oil that was used without further

r r purification.

¹ H NMR (CDCl ₃ ): δ 7.48-7.18 (8H, m), 4.42 (2H, s), 2.38 (3H, s).

The title compound was prepared in a manner analogous to that in Example C (1). 6-Isothiocyanato-1H-benzoimidazole (see Boev et al., Phar, Chem. J., vol. 24 (1990), pp. 812-822) and 2-bromoacetyl-3-methylbiphenyl gave yellow solid in 49% yield, mp Mp 184-190 ° C.

¹ H NMR (DMSO-d 6): δ 8.13 (1H, s), 7.87 (IH, s), 7.53 (1H, d, J = 8.7 Hz), 7.46-7, 39 (2H, m), 7.38-7.15 (7H, m), 2.35 (3H, s). HRFABMS (M +): Calcd .: 42.3, 1389. Found: 426.1374.

Anal. calculated for C ₂₄ H ₁₉ N ₅ OS. 1.0 H ₂ .0.3 CH ₃ CN: C, 64.82; H, 4.84; N, 16.29; S, 7.03. Found: C, 64.88; H, 4.69; N, 16.40;

S, 7.28.

* Example C (68):

(4-Amino-2- (4-methoxy-benzylamino) -thiazol-5-yl) - (2,5-dimethylthiophen-3-yl) -methanone

The title compound was prepared in a manner similar to that in Example C (1).

3-Bromoacetyl-2,5-dimethyl-thiophene (from Example C (52)) a

1- (2-isothiocyanato-ethyl) -4-methoxy-benzene gave a white solid in 72% yield, m.p. 175 ° C.

Ή NMR (DMSO-d 6): δ 6.68 (2H, d, J = 8.7 Hz), 6.74 (2H, d, J = 8.7 Hz), 6.41 (1H, s), 6.24 (1H, s), 4.88 (2H, s), 3.78 (3H, s), 2.40 (3H, s), 1.98 (3H, s).

IR (KBr): 3311, 2920, 1663, 1552, 1514, 1244 cm &^lt; -1 & gt ^; .

FABMS (MH &lt; + &gt;): 380 cm &lt; -1 &gt;

Anal. Calcd. for C ₁₈ H ₁₉ N ₃ O ₂ S ₂ : C, 57.88; H, 5.13; N, 11.25; S, 17.17. Found: C, 57.97; H, 5.11; N. 11.53; S, 17.28.

* Example C (69):

(4-Amino-2- (4-morpholin-4-yl-phenylamino) -thiazol-5-yl) - (3,5-dichloropyridin-4-yl) -methanone

The title compound was prepared in a manner analogous to that used in Example C (1).

4- (4-Isothiocyanato-phenyl) -morpholine (from Example C (54)) and 4-bromoacetyl-3,5-dichloropyridine (from Example C (63)) gave a yellow solid in 58% yield, m.p. 291.5-292.5 ° C.

1 H NMR (DMSO-d ₆ ): δ 10.75 (2H, s), 8.71 (2H, s), 8.32 (1H, s), 8.01 (1H, bs), 7.30 (2H, bs), 6.92 (2H, d, J = 9.0 Hz), 3.70 (4H, t, J = 4.5 Hz), 3.05 (4H, t, J = 4.5) Hz).

FABMS (MH &lt; + &gt;): 450/452

Anal. calcd for C ₁₇ H _{X 9} N ₅ O ₂ SC1 _2: C, 50.67; H, 3.80; N, 15.55;

S, 7.12, Cl, 15.74. Found: C, 50.55; H, 3.83; N, 12.59; WITH,

6.95; Cl, 15.47.

* Example C (70):

[4-Amino-2- [4- (4-methyl-piperazin-1-yl) -phenylamino] -thiazol-5-yl] - (3,5-dichloro-pyridin-4-yl) -methanone h ₂ h ₃ c _.n ^ </ ^N .Cl • s

C1-Methyl-4- (4-nitrophenyl) -piperazine, which has the structural formula:

/ ^N ~ • no ₂ er was prepared first. A mixture of 1-methyl-piperazine (4.00 g, 39.9 mmol) and 1-chloro-4-nitrobenzene (3.14 g, 20.0 mmol) was heated at 80 ° C for 24 hours then allowed to cool. and was diluted with water. The aqueous layer was extracted with MeOH: CH ₂ Cl ₂ (20:80; 4x50 mL). The combined organic layers were combined over MgSO ₄ , filtered, concentrated under reduced pressure, and recrystallized from ethanol to give 3.2 g (75% yield) of a yellow solid, which corresponded to the material previously described by ³ H NMR (de Silva et al. (J. Chem. Soc., Perkin Trans. 2, Vol. 9 (1993), pp. 1611-1616) and used without further purification.

4- (4-Methyl-piperazin-1-yl) aniline, with structural formula:

it was then prepared as follows. To a suspension of 1-methyl-4- (4-nitrophenyl) piperazine (2 g, 9.02 mmol) in absolute ethanol (30 mL) was added 10% Pd-C (250 mg). The resulting mixture was stirred under a H ₂ atmosphere for 5 hours, then filtered through a pad of celite. The filtrate was concentrated under reduced pressure to give 1.7 g (99% yield) of a brown solid which was used without further purification.

Έ NMR (CDCl ₃ ): δ 6.81 (2H, d, J = 8.8 Hz), 6.62 (2H, d, J = 8.8 Hz), 3.42 (2H, bs), 3 15 (4H, t, J = 5.0 Hz), 2.68 (4H, t, J = 5.0 Hz), 2.40 (3H, s).

1- (4-Isothiocyanato-phenyl) -4-methylpiperazine having the structural formula:

was prepared by a procedure analogous to that of

1- (4-isothiocyanato-phenyl) -1H-imidazole from Example C (41).

4- (4-Methyl-piperazin-1-yl) -aniline gave 1.7 g (83% yield) of a cream colored solid. m.p. Mp 118-120 ° C. (lit. 120-122 ° C, Galstuckova et al., J. Org. Chem. USSR (English translation), vol.

(1969), pp. 1121-1124), which was used without further purification. The IR spectrum corresponded to that of Martvon et al., Chem. Rumors, Vol. 27 (1973), pp. 808-810.

¹ H NMR (CDCl ₃ ): δ 7.20 (2H, d, J = 9.0 Hz), 6.82 (2H, d, J = 9.0 Hz), 3.20 (4H, dd, J = 5.0, 4.7 Hz), 2.52 (4H, dd, J = 5.0, 4.7 Hz), 2.24 (3H, s).

Anal. Calcd. for C ₁₂ H ₁₅ N ₃ S: C, 61.77; H, 6.48; N, 18.01; S, 12.69. Found: C, 61.51; H, 6.56; N, 17.86; S, 13.69.

The title compound was prepared by a procedure similar to that described in Example C (1). 1- (4-Isothiocyanatophenyl) -4-methyl-piperazine and 4-bromoacetyl-3,5-dichloro-pyridine (from Example C (63)) gave the crude product which, after recrystallization with EtOH / H ₂ O, afforded 40 mg ( 23% yield) pale brown solid, mp 150-151 ° C.

1 H NMR (DMSO-d ₆ ): δ 10.78 (1H, s), 8.70 (IH, s), 8.00-8.41 (2H, m), 7.24 (2H, bs) 6.88 (2H, d, J = 9.0 Hz), 3.08 (4H, dd, J% 5.0,

4.7 Hz), 2.40 (4H, dd, J = 5.0, 4.7 Hz), 2.20 (3H, s).

IR (KBr): 3395, 2925, 1618, 1546, 1514, 1426, 1240 ^cm-first

HRFABMS: Calcd for C ₂₀ H ₂₁ Cl ₂ N ₅ OS (MH +): 463.0875. Found: 563.0861.

Anal. calculated for C ₂₀ H ₂₀ N ₆ OSCl ₂ . 0.6 H ₂ O. 0.1 EtOH. 0.05 CHC1 _3: C, 50.20; H, 5.06; N, 16.22; S, 6.19, Cl, 14.71. Found: C, 50.34; H, 5.11; N, 16.53; S, 6.43; Cl, 14.74.

* Example C (71):

[4-Amino-2- [4- (4-methyl-piperazin-1-yl) -phenylamino] -thiazol-5 - [(2,6-dichloro-phenyl)] -methanone

e r

The title compound was prepared as described in Example C (1). 1- (4-isothiocyanato-phenyl) -4-methyl-piperazine (from Example C (70)) and 2-bromo-2 ', 6'-dichloro-acetophenone (from Example G (52)) gave, after recrystallization with H ₂ O / EtOH / CH ₂ Cl ₂ , 2.2 g (64% yield) of a yellow solid. mp 160-162 ° C.

¹ H NMR (DMSO-d 6): δ 10.60 (1H, s), 8.00 (2H, bs), 7.20-7.41 (4H, m), 6.88 (2H, d, J = 9.0 Hz), 3.08 (4H, dd, J = 5.0, 4.7 Hz), 2.40 (4H, dd, J = 5.0, 4.7 Hz), 2.18 ( 3H, s).

IR (KBr): 3394, 3164, 2942, 2810, 1610, 1546, 1427, 1242 cm &^lt; -1 & gt ^; .

HRFABMS: Calcd for C ₂₁ H ₂₂ Cl ₂ N ₅ OS (MH +): 462.0922. Found: 462.0906.

Anal. calculated for C ₂₁ H ₂₁ N ₅ OSCl ₂ . .0.5 H ₂ 0. 1 EtOH. 0.1 CH ₂ C1 _2: C, 52.75; H, 5.40; N, 13.32; S, 6.10, Cl, 14.83. Found: C, 53.06; H, 5.37; N, 13.51; S, 6.26; Cl, 14.63.

* Example C (72):

4-Amino-2- (1H-benzoimidazol-6-ylamino) thiazol-5-yl- (3,5-dibromothiophen-2-yl) methanone

Br,

Br

2-Acetyl-3,5-dibromothiophene, with structural formula:

was prepared first as follows. To a solution of 2,4-dibromothiophene (2.0 g, 8.27 mmol) and acetyl chloride (0.82 mL, 11.6 mmol) in ether (3 mL) was added portionwise AlCl ₃ (1.5 g,

11.2 mmol). After 4 hours, an additional portion of acetyl chloride a was added

AlCl ₃ , the mixture was refluxed for 1 hour and allowed to cool.

The reaction mixture was carefully cooled with ice and extracted with ether.

The ether layers were decolourised with activated carbon, dried over MgSO ₄ , passed through a pad of silica gel and concentrated to give

err eerr re .- · - rrrrr, Γ ·· - rrrrrrrr 'rrr ~ r - ~ _r c

I, 8 g (77% yield) of a dark brown oil which was ^X H-NMR spectrum corresponding to the above described, see del Agua et al., J. Heterocycl. Chem., Vol. 18 (1981) p. 1345-1347, and was used without further characterization.

2-Bromoacetyl-3,5-dibromothiophene having the structural formula:

was then prepared as described in analogy to 2-bromo-2'-iodoacetophenone, see Example C (12).

2-Acetyl-3,5-dibromo-thiophene (220 mg, 0.77 mmol) yielded 295 mg of a dark brown solid which was used without further purification.

¹ H NMR (CDCl ₃ ): δ7.13 (1H, s), 4.54 (2H, s).

Finally, the title compound was prepared in a manner analogous to that used in Example C (1). 6-Isothiocyanato-1H-benzoimidazole (see Boev et al., Pharm. Chem.

J. (English translation), vol. 24 (1990), pp 812-822), and

2-bromoacetyl-3,5-dibromothiophene gave a dark brown solid in 50% yield, m.p. Mp 261-264 ° C.

1 H NMR (DMSO-d ₅ ): δ 12.50 (1H, bs), 10.94 (2H, s), 8.27 (2H, bs), 8.21 (1H, s), 7, 87 (1H, bs), 7.57 (1H, d, J = 8.7Hz), 7.36 (1H, s), 7.24 (1H, d, J = 8.7).

HRFABMS (MH +): Calcd .: 499.8673. Found: 499.8686.

Anal. calculated for C ₁₅ H ₉ N ₅ OS ₂ Br ₂ . 0.5 H ₂ O: C, 35.45; H, 1.98; N, 13.78; S, 12.62; Br, 31.45. Found: C, 35.37; H, 1.73; N,

13.52; S, 12.75; Br ,. 31.25.

* Example C (73):

4- [4-Amino-5- (3,5-dibromo-thiophene-2-carbonyl) -thiazol-2-ylamino] -benzenesulfonamide

The title compound was prepared by a procedure analogous to that used in Example C (1).

4-Isothiocyanato-benzenesulfonamide and 2-bromoacetyl-3,5-dibromothiophene (from Example C (72)) gave a yellow powder in 41% yield, m.p. Mp 254-255 ° C.

NMR (DMSO-d6): δ 11.24 (1H, s), 8.31 (2H, bs), 7.77 (4H, s), 7.40 (1H, s), 7.28 (2H, s).

FABMS (MH &lt; + &gt;): 539/538/540.

Anal. Calcd for C ₁₄ H ₁₀ N ₄ O ₃ S ₃ Br ₂ : C, 31.24; H, 1.87; N, 10.41; S, 17.87; Br, 29.69. Found: C, 31.08; H, 1.90; N, 10.16; S, 17.69; Br, 29, 96.

* Example C (74):

/ 4-Amino-2- (lH-benzoimidazol-6-ylamino) -thiazol-5-yl / - (1,5-dimethyl-lH-imidazol-4-yl) -methanone NH _2; h ₃

N '

H ^N ^ ^{N '} _CH3

1,5-Dimethyl-lH-imidazole-4-carboxylic acid having the structural formula: _Ln ³ him> CH ₃ was prepared by first following cspostupem. A fresh solution of NaOH (3.86 g, 96.5 mmol) in water (20 mL) was added to. a solution of ethyl 1,5-dimethyl-1H-imidazole-4-carboxylate (5.39 g,

32.0 mmol); Ohno et al., Chem. Pharm. Bull., Vol. 42 (1994), pp. 1463-1473) in EtOH (20 mL). After 5 hours, the mixture was cooled to 0 ° C and acidified with 38% HCl to pH 3-4. The solid obtained was filtered, washed with a small amount of cold EtOH: H ₂ O (1: 1), and dried under high vacuum to give 3.51 g (78%) of a white solid which was used without further purification.

¹ H NMR (D ₂ O): δ 8.49 (1H, s), 3.73 (3H, s), 2.46 (3H, s).

Γ Γ

Anal. Calcd. for C ₆ H ₈ N ₂ O ₂ : C, 51.42; H, 5.75; N, 19.99. Found: C, 51.52; H, 5.78; N, 19.98.

1,5-Dimethyl-1H-imidazole-4-carboxylic acid N-methoxy-N-methylamide having the structural formula: ³

was then prepared as follows: Into the mixture

1,5-dimethyl-1H-imidazole-4-carboxylic acid (2.01 g, 14.4 mmol) in DMF (20 mL) was added O- (7-azabenzotriazol-1-yl) -Ν, Ν, Ν ,, Ν'-tetramethyluronium hexafluorophosphate (HATU; 6.00 g, 15.8 mmol) and diisopropylethylamine (7.5 mL, 43 mmol). After 5 minutes, N, O-dimethylhydroxylamine hydrochloride (1.54 g, 15.79 mmol) was added to the resulting clear solution. After 1 hour, the resulting yellow solution was partitioned between CHCl ₃ and water. The separated organic layer was washed with water, and brine, dried over K ₂ CO ₃ , concentrated and dried under high vacuum to give 1.88 g (72% yield) of a light brown solid which was used without further purification.

¹ H NMR (CDCl ₃ ): δ 7.36 (1H, s), 3.81 (3H, s), 3.56 (3H, s), 3.47 (3H, s), 2.45 (3H) , s).

1- (1,5-Dimethyl-1H-imidazol-4-yl) ethanone having the structural formula:

was then prepared as follows: To a solution of crude 1,5-dimethyl-1H-imidazole-4-carboxylic acid N-methoxy-N-methylamide (1.69 g, 9.21 mmol) in THF (55 mL) at -78 ° C was added dropwise 1.4M CH ₃ MgBr in ether (8.55 mL, 12.0 mmol). The mixture was allowed to warm to ambient temperature over one hour, then quenched with 1N HCl, adjusted to basic pH 9 with 1N NaOH, concentrated under reduced pressure to remove THF, and extracted with EtOAc (200 mL). The organic layer was separated, dried over K ₂ CO ₃ , and evaporated to give 1.2 g (94% yield) of a yellow solid which was used without further purification.

Ή NMR (CDCl ₃ ): δ 7.35 (1H, s), 3.57 (3H, s), 2.55 (3H, s), 2.53 (3H, s).

2-Bromo-l- (1,5-dimethyl-lH-imidazol-4-yl) -ethanone, which has the structural formula: ^H 3 <?

was further deprived of the following procedure.

To 1- (1,5-dimethyl-1H-imidazol-4-yl) ethanone (464 mg, 3.36 mmol) in HOAc (8.5 mL) was added dropwise bromine (173 μ 3, 3) at 0 ° C. , 36 mmol). After 36 hours at ambient temperature, the crude 2-bromo-1- (1,5-dimethyl-1H-imidazol-4-yl) ethanone hydrobromide salt was filtered off as a brown solid, which was subsequently washed with a minimum amount of water and ether, dissolved in CHCl ₃ , cooled to 0 ° C, treated with NaHCO ₃ and concentrated under reduced pressure below 40 ° C to give 719 mg (99% yield) of a yellow oil which was used without further purification.

1 H NMR (DMSO-d ₅ ): δ 8.40 (1H, s), 4.68 (2H, s), 3.66 (3H, s), 2.67 (3H, s).

The title compound was prepared in a manner analogous to that used in Example C (1). 6-Isothiocyanate-1H-benzoimidazole (see Boev et al., Pharm. Chem. J. (Eng. Trans.), Vol. 24 (1990), pp. 812-822) and 2-bromo-1- (1, 2-bromo-1-carboxylic acid) 5-dimethyl-1H-imidazol-4-yl) ethanone gave a dark brown solid in 15% yield, m.p. 275.5-277 ° C.

Ή NMR (DMSO-d ₆ ): δ 12.42 (1H, s), 10.42 (1H, s), 8.16 (1H, s), 7.94 (1H, s), 7.61- 7.30 (2H, m), 7.26 (1H, dd, J = 8.4, 1.9 Hz), 3.54 (3H, s), 2.51 (3H, s).

HRFABMS (MH +): Calcd. 354.1137. Found: 354.1132.

Anal. calculated for C ₁₆ H ₁₅ N ₇ OS. 0.5 H ₂ 0. 0.8 CH ₃ OH: C, 52.00; H, 4.99; N, 25.27; S, 8.26. Found: C, 52.27; H, 4.81; N, 25.06; S, 8.12.

* Example C (75):

(4-Amino-2- (4-morpholin-4-yl-phenylamino) -thiazol-5-yl) - (2,6-dichloro-3-nitro-phenyl) -methanone

2-Bromo-2 formula:

6'-dichloro-3'-nitro-acetophenone having a structural α

Br— ⁷ \ == /

Cl _{2 of a} solution of 2 '', 6'-dichloro-3 Breslin et al., J. Med. glacial acetic acid was prepared as follows. To -nitro-acetophenone (1.3 g, 5.6 mmol;

Chem., Vol. 38 (1995), pp. 791-793) in (5 ml) bromine (325 μΐ, 6.83 mmol) was added at ambient temperature. The resulting mixture was warmed to 80 ° C for 1 hour, allowed to cool, and diluted with ether. The organic layer was washed with ice-cold H ₂ O (25 mL), saturated aqueous NaHCO ₃ (3x25 mL), and brine (25 mL), dried over MgSO ₄ and concentrated under reduced pressure to give 1.7 g (97% in crude) yield) of a yellow oil which was used without further purification.

Ή NMR (CDCl ₃ ): δ 7.98 (1H, d, J = 8.7 Hz), 7.38 (1H, d, J = 8.7 Hz), 4.40 (2H, s).

The title compound was prepared in a manner similar to that described in Example C (1). 2-Bromo-2 ', 6'-dichloro-3'-nitroacetophenone and 4- (4-isothiocyanato-phenyl) morpholine (from Example C (54)) gave a solid raw material which, after purification by flash column chromatography with hexane / EtOAc ( 70:30) as eluent gave a dark brown foam in 52% yield, mp 170-172 [deg.] C. .

¹ H NMR (DMSO-d ₆ ): δ 10.70 (1H, s), 8.30 (1H, s), 8.10 (1H, d, J = 9.0 Hz), 7.90 (1H δ d, J = 8.7 Hz), 7.20-7.30 (2H, m), 6.90 (2H, d, J = 9.0 Hz), 3.70 (4H, dd, J = 5.0, 4.7 Hz), 3.06 (4H, dd, J = 5.0, 4.7 Hz).

IR (KBr): 3289, 2966, 2848, 1634, 1542, 1425, 1343, 1225, 1108 cm &lt; ^{-1 &gt};.

HRFABMS: Calcd. for C ₂₀ H ₁₇ Cl ₂ N ₅ O ₄ SNa (M + Na ⁺ ): 516.0276.

Found: 516.0258.

Anal. calc. for C ₂₀ H ₁₇ Cl ₂ N ₅ O ₄ S. 0.35 CHC1 _3: C, 45.59; H, 3.26; N, 13.06; S, 5.98; Cl, 20.27. Found: C, 45.33; H, 3.37; N, 12.96; S, 5.93; Cl, 20.27.

* Example C (76):

4- / 4-Amino-5- (1,5-dimethyl-1H-imidazol-4-carbonyl) -thiazol-2-ylamino / benzenesulfonamide

N ^ ^N-CH ₃

The title compound was prepared in a manner analogous to that used in Example C (1). 4-Isothiocyanato-benzenesulfonamide and 5-bromoacetyl-1,5-dimethyl-1H-imidazole (from Example C (74)) gave a yellow solid in 8% yield, m.p. Mp 293-294 ° C.

1 H NMR (DMSO-d ₅ ): δ 10.80 (1H, s), 7.81 (2H, d, J = 9.0 Hz), 7.75 (2H, d, J = 9.0 Hz) ), 7.62 (1H, s), 7.24 (2H, s), 3.56 (3H, s), 2.52 (3H, s).

HRFABMS (M + Na ⁺ ): Calcd. : 415,0623. Found: 415.0609.

Anal. calc. for C ₁₅ H ₁₆ N ₆ O ₃ S ₂ · 1/0 CH ₃ OH. 1.0 CHC1 _3: C, 42.53; H, 4.45; N, 18.26; S, 13.93.

Found: C, 42.57; H, 4.41; N, 18.18; S, 14.07.

* Example C (77):

[4-Amino-2- (4-morpholin-4-yl-phenylamino) -thiazol-5-yl] - (1,5-dimethyl-1H-imidazol-4-yl) -methanone

The title compound was prepared in an analogous manner to that used in Example C (1). 4- (4-Isothiocyanato-phenyl) morpholine (from Example C (54)) and 5-bromoacetyl-1,5-dimethyl-1H-imidazole (from Example C (57)) gave a yellow solid in 12% yield, m.p. higher than 300 ° C.

¹ H NMR (DMSO-d ₆ ): δ 10.21 (1H, s), 7.57 (IH, s), 7.42 (2H, d, J = 8.8 Hz), 6.94 (2H δ d, J = 8.8 Hz), 3.72 (4H, t, J = 4.7 Hz), 3.54 (3H, s), 3.06 (4H, t, J = 4.7 Hz) 2.50 (3H, s).

HRFABMS (M +): Calcd .: 398.1525. Found: 398.1516.

Anal. calc. for C ₁₉ H ₂₂ N _with O ₂ S. 0.2 CH ₃ OH. 0.2 CHC1 _3: C, 54.53; H, 5.41; N, 19.60; S, 7.48.

Found: C, 54.63; H, 5.27; N, 19.56; S, 7.47.

* Example C (78):

4-Amino-2- (1H-benzoimidazol-6-ylamino) thiazol-5-yl-7- (3-methyl-5-nitrothiophen-2-yl) methanone

2-Acetyl-3-methyl-5-nitrothiophene with structural formula: was prepared first as follows.

2-Bromo-3-methyl-5-nitrophen (5.17 g, 23.3 mmol; Spinelli et al.,

Γ I “! * Γ r Γ rrcr 9. ·· rr ^{Γ Γ f} . 2? · - - ^l

J. Chem. Soc.Perkin Trans. 2, (1975), pp. 620-622), tributyl (1-ethoxyvinyl) tin (8.65 mL, 25.6 mmol), and palladium dichlorobis (triphenylphosphine) (163 mg, 0.23 mmol) in toluene (10.5 mL) were heated under Ar at 100 ° C for 2.5 hours. 5% aqueous HCl (78 mL) was added, and the mixture was stirred for 15 min at 60 ° C, then partitioned with ether and water. The organic layer was separated, dried over MgSO ₄ and concentrated to a residue that was dissolved in ether (130 mL).

1,8-Diazabicyclo [5.4.0] undec-7-ene (DBU; 2.2 eq.) And a 0.1 M iodine solution in ether were added dropwise until the color persisted for several seconds. The resulting solution was passed through a short silica gel column and concentrated in vacuo to give 3.74 g (87% yield) of a yellow solid, which was used without further purification.

¹ H NMR (CDCl ₃ ): δ 7.71 (1H, s), 2.58 (3H, s), 2.57 (3H, s).

2-bromoacetyl-3-methyl-5-nitro-thiophene, which has the structural formula H ₃ C N ^Br <X _N0 of ^N ° ² was prepared in a manner analogous to 2-bromo-2'-iodo-acetophenone, see Example C (12). 2-Acetyl-3-methyl-5-nitro-thiophene (230 mg, 1.24 mmol) yielded 330 mg of a dull yellow oil containing traces of a bromine by-product as determined by NMR, which was used without further purification.

¹ H NMR (CDCl 3): δ 7.75 (1H, s), 4.28 (2H, s), 2.60 (2H, s).

The title compound was prepared in a manner analogous to that used in Example C (1). 6-Isothiocyanato-1H-benzoimidazole (see Boev et al., Phar. Chem.J. (English)), Vol. 24 (1990), pp. 818-822) and 2-bromoacetyl-3-methyl-5 -nitro-thiophene gave a yellow solid in 23% yield, mp above 300 ° C.

¹ H NMR (DMSO-d ₆ ): δ 12.51 (1H, d, J = 14.3 Hz), 11.01 (1H, bs), rr

8.40 (2H, bs), 8.21 (1H, s), 8.02 (1H, s), 7.63 (1H, bs), 7.52 (1H, bs), 7.36 (1H) , d, J = 11.0 Hz), 2.33 (3H, s).

HRFABMS (M +): Calcd .: 401.0491. Found: 401.0474.

Anal. calc. for C _ls H ₁₂ N ₆ O ₃ S _2nd 0.7 H ₂ O. 0.8 CH ₃ OH: C, 46.00; H, 3.81; N, 19.16; S, 14.62.

Found: C, 45.92; H, 3.50; N, 19.096; S, 15.49.

* Example C (79):

(4-Amino-2- (1H-benzoimidazol-6-ylamino) thiazol-5-yl) - (2,6-difluorophenyl) methanone

2-Bromo-2 ', 6'-difluoroacetophenone having the structural formula was prepared as a first procedure analogous to the procedure for the preparation of 2-bromo-2'-iodoacetophenone, see Example C (12).

2 ', 6'-difluoroacetophenone (703 mg, 4.5 mmol) gave 1.01 g (96% yield) of a pale yellow oil which was used without further purification.

¹ H NMR (CDCl ₃ ): δ 7.56 - 7.42 (1H, m), 7.07 - 6.98 (7.42 (2H, d, J = 2H, m), 4.38 (2H) , s).

The title compound was prepared in a manner analogous to that used in Example C (1). 6-Isothiocyanato-1H-benzoimidazole (see Boev et al., Phar. Chem.J., vol. 24 (1990), pp. 818-822) and 2-bromo-2 ', 6' -difluoroacetophenone gave yellow solid crystals in 78% yield, mp Mp 194-200 ° C.

¹ H NMR (DMSO-d ₆ ): δ 12.45 (1H, s), 10.86 (1H, s), 8.19 (1H, s),

8.16 (2H, bs), 7.80 (1H, bs), 7.59-7.44 (2H, m), 7.22-7.11 (3H, m).

r ··

C * * r t- yy yy yy yy <“f yy yy y

HRFABMS (MH +): Calcd .: 372.0731. Found :. 372.0725.

Anal. calc. for CpHnNsOSFj. 0.5 H ₂ O: C, 53.86; H, 3.18; N, 18.41; S, 8.43.

Found: C, 53.73; H, 3.14; N, 18.32; S, 8.53.

* Example C (80):

(4-Amino-2- [4- (4-methyl-piperazin-1-yl) -phenylamino] -thiazol-5-yl) - (2,6-difluoro-phenyl) -methanone imh ₂

The title compound was prepared in a manner analogous to that used in Example C (1).

1- (4-Isothiocyanato-phenyl) 4-methyl-piperazine (from Example C (70)) and 2-bromo-2 ', 6'-difluoroacetophenone (from Example C (79)) gave a yellow solid in 71 % yield, mp Mp 168-170 ° C.

¹ H NMR (DMSO-d 6): δ 10.62 (1H, s), 8.11 (2H, bs), 7.54-7.43 (1H, m), 7.28 (2H, d, J) = 7.5 Hz), 7.20-7.10 (2H, m), 6.90 (2H, d, J = 9.0 Hz), 3.08 (4H, t, J = 4.8 Hz) 1.41 (4H, t, J = 4.8 Hz), 2.19 (3H, s).

IR (KBr): 2942, 2809, 1620, 1590, 1546, 1516, 1464, 1429, 1238, 1002 cm ^-1 .

HRFABMS (MH +): Calcd .: 430.1513. Found: 430.1502.

Anal. calc. for C ₂₁ H ₂₁ N ₅ OSF ₂ . 0.3 H ₂ O: C, 58.00; H, 5.01; N,

16.10; S, 7.37.

Found: C, 57.98; H, 4.92; N, 16.08; S, 7.42.

* Example C (81):

((4-Amino-2- [4- (4-methyl-piperazin-1-yl) -phenylamino) -4-thiazol-5-yl) - (2,6-dichloro-4-trifluoromethyl-phenyl) -methanone o

HjC — N

The title compound was prepared in a manner similar to that in Example C (1). 1- (4-Isothiocyanato-phenyl) -4-methyl-piperazine (from Example C (70)) and 2-bromo-2 ', 6'-dichloro-4'-trifluoromethyl-acetophenone gave, after recrystallization from EtOAc / hexane, yellow needles in 68% yield, mp Mp 239-240 ° C.

¹ H NMR (DMSO-d 6): δ 8.00 (2H, s), 7.28 (2H, bs), 6.91 (2H, d, J = 8.7 Hz), 3.10 (4H, dd) J = 5.1, 4.7 Hz), 2.42 (4H, dd, J = 5.1, 4.8 Hz), 2.20 (3H, s).

IR (KBr): 3377, 3283, 2942, 2831, 1598, 1542, 1513, 1425 cm &^lt; -1 & gt ^; . FABMS (M + Na &lt; + &gt;): 552.

Anal. calculated for C ₂₂ H ₂₀ Cl ₂ F ₃ N ₅ OS. H, O. 0.7 C ₆ H _14: C, 52.00; H, 5.23; N, 11.57; S, 5.30, Cl, 11.72.

Found: C, 51.94; H, 4.98; N, 11.18; S, 5.20; Cl, 11.48.

* Example C (82):

N- [3- [1-Amino-2- (1H-benzoimidazol-5-ylamino) -thiazole-5-carbonyl] -2,4-dichlorophenyl] acetamide

3'-Amino-2 ', 6'-dichloroacetophenone having the structural formula ^{Cl NH 2} was prepared by the first procedure. To a solution of SnCl ₂ .2H ₂ O (7.70 g, 34.2 mmol) in 6N aqueous HCl (20 mL) was added

2 ', 6'-dichloro-3'-nitroacetophenone (4.00 g, 17.1 mmol; Breslin et al., J. Med. Chem., Vol. 38 (1995)), pp. 791-793). The resulting mixture of f Γ f ϊ was heated under reflux for 5 hours, allowed to cool and was carefully treated with anhydrous Na ₂ CO ₃ . The resulting white precipitate was filtered and washed with CHC1 _third The organic layer was sealed and the aqueous layer was extracted with CHCl ₃ (3x50 mL). The combined organic layers were dried over MgSO ₄ , filtered, and concentrated in vacuo to give a black oil which was purified by flash column chromatography with EtOAc: hexane (20:80) as eluent. 2.6 g (75% yield) of a pale brown oil were obtained, which was used without further purification.

¹ H NMR (CDCl ₃ ): δ 7.08 (1H, d, J = 8.7 Hz), 6.70 (IH, d, J = 8.7 Hz), 4.12 (2H, bs), 2.56 (3 H, s).

N- (3-Acetyl-2,4-dichlorophenyl) acetamide having the structural formula

, was prepared subsequently. Into solution

3-Amino-2 ', 6'-dichloroacetophenone (2.40 g, 11.8 mmol) in glacial acetic acid (25 mL) was added acetic anhydride (5.53 mL, 58.8 mmol). The resulting mixture was heated under reflux for 2 hours, allowed to cool and diluted with ether (100 mL). The organic layer was washed with H ₂ O (2 x 50 mL), dried over MgSO ₄ , concentrated in vacuo and azeotroped with n-heptane to give 2.3 g of a pale white solid which was used without further purification.

¹ H NMR (CDCl ₃ ): δ 8.38 (1H, d, J = 9.1 Hz), 7.62 (IH, bs), 7.34 (IH, d, J = 9.0 Hz), 2.60 (3H, s); 2.22 (3H, s).

N- (3-Bromoacetyl-2,4-dichlorophenyl) acetamide having the structural formula

Ck [[N ₃ O ₃ or ^{H 2} ], was prepared in a manner analogous to that of

2-bromo-2 ', 6'-dichloro-3'-nitroacetophenone for Example C (75).

r- f r f »· r r r r

Γ f f r c c r C r c

'

N- (3-Acetyl-2,4-dichlorophenyl) -acetamide gave a pale brown oil in 100% crude yield, which was used without further purification.

¹ H NMR (CDCl ₃ ): δ 8 _Z 48 (1H, d, J = 8.7 Hz), 7.60 (1H, bs), 7.38 (1H, d, J = 9.0 Hz), 4.40 (2H, s); 2.2 (3H, s).

The title compound was prepared as described for Example C (1). 6-Isothiocyanato-1H-benzoimidazole (see Boev et al., Pharm. Chem. J. (English translation)., Vol. 24 (1990), pp. 818-822) and N- (3-bromoacetyl-2, 3-bromoacetyl). 4-dichlorophenyl) acetamide gave the product, which was purified by flash chroamtography with a step gradient of MeOH: CH ₂ Cl ₂ (10:90) to HOAc: MeOH: CH ₂ Cl ₂ (1:10:90) to give a yellow foam in 56% yield which decomposed above 200 ° C.

¹ H NMR (DMSO-d 6): δ 9.90 (1H, s) -8.20 (1H, s), 7.84-7.96 (1H, m) 7.68 (1H, d, J = 7.4 Hz), 7.58 (1H, d, J = 8.8 Hz), 7.24 (1H, d,

J = 8.4 Hz), 2.20 (3H, sj.

IR (KBr): 3295, 1625, 1525, 1425 cm &^lt; -1 & gt ^; .

HRFABMS. Calcd (MH ⁺ ,: 461.0354. Found: 461.0344).

Anal. calculated for C ₁₉ H ₁₅ Cl ₂ N ₆ O ₂ S. H ₂ O. H, 4.28; N, 12.74; S, 4.86; Cl, 10.75.

Found: C, 45.93; H, 4.08; N, 12.49; S, 4.83; Cl, 10.45.

rr * Example C (83):

4-Amino-2- (4-morpholin-4-yl-phenylamino) -thiazol-5-yl-7- (3-methyl-biphenyl-2-yl) -methanone

The title compound was prepared in a manner similar to that in Example C (1).

4- (4-Isothiocyanato-phenyl) -morpholine (from Example C (54)) and 2-bromoacetyl-3-methyl-biphenyl (from Example C (67) gave a yellow solid in 29% yield, mp 125-35 ° C).

¹ H NMR (DMSO-d 6): δ 10.40 (1H, s), 7.86 (2H, s), 7.42-7.24 (9H, d, J = 7.5 Hz), 6, 93 (2H, d, J = 8.7Hz), 3.73 (4H, t, J = 4.4Hz), 3.07 (4H, t, J = 4.4Hz), 2.26 ( 3H, s).

HRFABMS (M): Calcd .: 471.1855. Found: 471.1839.

Anal. calculated for C ₂₇ H ₂₆ N ₄ O ₂ S. 1.0 CF ₃ CO ₂ H: C / 59.58; H, 4.66; N, 9.58; S, 5.48. Found: C, 59.41; H, 5.01; N, 9.26; S, 5.18.

r ·· *

000 r · 0

0 r 0 r r 0cr ♦ Example C (84):

(4-Amino-2- (4-morpholin-4-yl-phenylamino) -thiazol-5-yl) - (2-bromo-6-methyl-phenyl) -methanone

The title compound was prepared in a manner similar to that in Example C (1).

4- (4-Isothiocyanato-phenyl) -morpholine (from Example C (54)) a

2,2'-Dibromo-6'-methylacetophenone (from Example C (66)) gave a crude solid that was triturated with MeOH / CHCl to give a yellow solid in 22% yield, mp 105-125 ° C.

1 H NMR (DMSO-d 6): δ 10.57 (1H, s), 8.01 (2H, bs), 7.46 (1H, d, J = 7.5 Hz), 7.39-7.18 (4H, m), 6.96 (2H, d, J = 8.7 Hz), 3.74 (4H, t, J = 4.7 Hz), 3.09 (4H, t, J = 4, 7 Hz), 2.20 (3H, s).

HRFABMS (MH ⁺ ): Calcd .: 73.0664 / 475. Found: 473.0657 / 475.

Anal. calculated for C ₂₁ H ₂₁ N ₄ O ₂ SBr. 0.7 MeOH. 0.6 CHCl _; Found: C, 47.60; H, 4.37; N, 9.98; S, 5.71. Found: C, 47.95; H, 4.05; N, 9.77; S, 5.51.

♦ Example C (85):

4- (4-Amino-5- (2,6-difluoro-benzoyl) thiazol-2-ylamino) -benzenesulfonamide

The title compound was prepared in a manner similar to that in Example C (1). (4-Isothiocyanato-benzenesulfonamide and 2-bromo-2 ', 6'-difluoroacetophenob (from Example C (79)) gave pale yellow crystals in 69% yield, mp 258-260 ° C.

¹ H NMR (DMSO-d 6): δ 11.20 (1H, s), 8.20 (2H, bs), 7.79 (2H, d, J = 9.0 Hz), 7.74 (2H, d) J = 9.0 Hz), 7.61-7.49 (1H, m), 7.26 (2H, s), 7.22 (1H, d, J = 7.9 Hz), 7.19 (1 H, d, J = 8.0 Hz).

IR. (KBr): 3310, 1622, 1599, 1547, 1525, 1467, 425, 1410, 1318, 1156 cm - ^first

HRFABMS (MH ⁺ ) - Calcd .: 411.0397. Found: 411.0410.

Anal. calculated for C ₁₆ H ₁₂ N ₄ O ₃ S ₂ F ₂ . 0.7 CH ₃ OH: C, 46.34; H, 3.45; N, 12.94, S, 14.82. Found: C, 46.19; H, 3.12; N, 12.83; S, 14.94.

♦ Example C (86):

N- (3 - [(4-amino-2- [4- (4-methylpiperazin-1-yl) phenylamino) -thiazole-5-carbonyl] -2,4-dichlorophenyl) acetamide

The title compound was prepared in a manner similar to that in Example C (1). 1- (4-Isothiocyanato-phenyl) -4-methylpiperazine (from Example C (70)) and N- (2-bromoacetyl-3-chlorophenyl) acetamide (from Example C (82) yielded, after recrystallization with EtOH / CHCl ₃₎ 60 mg (13% yield) of a yellow solid, mp 195-197 ° C.

ΐ NMR (DMSO-d ₆ ): δ 10.62 (1H, s), 9.62 (1H, s), 7.90 (1H, bs), 7.78 (1H, dd, J = 8.9) 4.4 Hz), 7.47 (1H, d, J = 8.8 Hz), 7.30 (2H, bs), 6.92 (2H, d, J = 9.1 Hz), 3, Δ (4H, dd, J = 5.1, 4.6 Hz), 2.42 (4H, dd, J = 5.1, 4.6 Hz), 2.18 (3H, s), 2.08 (3 H, s). IR (KBr): 3260, 3025, 2801, 1666, 1613, 1525, 1437, 1382, 1299 cm - ^1st

100 ALIGN!

HRFABMS. Calcd. (M + Na ⁺ ): 541.0956. Found: 541, 0970.

Anal. calculated for C ₂₃ H ₂₄ Cl ₂ N ₆ O ₂ S. 0.5 H ₂ O. 0.4 EtOH: C, 52.27; H, 5.05; N, 15.37; S, 4.86, Cl, 12.97. Found: C, 52.13; H, 5.09; N, 15.13; H, 5.09; N, 15.13; S, 5.78; Cl, 12.96.

♦ Example C (87):

4-Amino-2- / 4- _(4-m ethyl-piperazin-l-yl) -fenylaminq / thiazole-5-yl / -3-methyl-thiophen-2-yl-methanone

The title compound was prepared in a manner similar to that in Example C (1). 1- (4-Xsothiocyanato-phenyl) 4-methylpiperazine (from Example C (70)) and 2-bromoacetyl-3-methylthiophene (from Example C (19) gave, after recrystallization with EtOH / CHCl, a dark yellow solid in 75% mp 237.0-237.5 ° C.

NMR (DMSO-d _6): δ 10.50 (1H, s), 8.10 (1H, bs), 7.56 (1H, d, J = 5.0 Hz), 7.38 (2H, d J = 8.8 Hz), 6.96 (3H, m), 3.10 (4H, dd, J = 5.1, 4.7 Hz), 2.45 (4H, dd, J = 4, 9, 4.7 Hz), 2.38 (3H, s),

2.24 (3 H, s).

IR (KBr): 3484, 3319, 2943, 2809, 1593, 1546, 1414 cm &^lt; -1 & gt ^; .

HRFABMS. Calcd. (*): 414.1422. Found: 414.1408.

Anal. calculated for C ₂₀ H ₂₃ N ₅ OS ₂ . 3 H ₂ O: C, 57.34; H, 5.68; N, 16.72 S, 15.31. Found: C, 57.01; H, 5.72; N, 16.41; S, 15.34.

♦ Example C (88):

trans-3RS-Amino-4RS - N - N -amino-5- (3,5-dichloropyridine-4-carbonyl)

thiazol-2-ylamino _from / -benzoyl / dihydrofuran-3-one rr <- ·

The title compound was prepared in a manner similar to that in Example C (1). 4-Isothiocyanato-benzoyl-DL-homoserine lactone and 4-bromoacetyl-3,5-dichloropyridine (from Example C (63)) gave the product which was purified by column chromatography with 10% MeOH / CHCl ₃ as eluent to give an amorphous yellow solid 203 mg (79%), which decomposed above 150 ° C.

¹ H NMR (DMSO-d ₆ ): δ 11.17 (1H, s), 8.89 (1H, d, J = 8.0 Hz), 8.76 (2H, s), 7.86 (2H δ d, J = 8.7 Hz), 7.69 (2H, d, J = 8.7 Hz), 4.73 (1H, q, J = 9.3 Hz), 4.42 (1H, ddd) J = 8.9, 8.7, 1.8 Hz), 4.27 (1H, ddd, J = 10.0, 8.7, 6.7 Hz).

HRFABMS. Calcd for C ₂₀ H ₁₅ Cl ₂ N _with O ₄ SNa (M + Na ⁺ ): Calcd .: 514.0120. Found: 514.0133.

IR (KBr): 3284, 1774, 1610, 1524, 1459, 1423, 1348, 1306, 1180 cm &lt; ^{-1 &gt};.

Anal. calculated for C ₂₀ H ₁₅ Cl ₂ N ₅ O ₄ S. 0.25 H ₂ O. 0.6 CHC1 _3: C, 43.52;

H, 2.85; N, 12.32; Cl, 23.70; S, 5.64. Found: C, 43.31; H, 2.78; N, 12.46; Cl, 24.07; S, 5.63.

♦ Example C (89):

N-amino-2- (1H-benzoimidazol-6-ylamino) -thiazol-5-yl] - (2,6-dichloro-3-nitrophenyl) methanone

102 ·

The title compound was prepared as described for Example C (1). 2-Bromo-2 ', 6'-dichloro-3'-nitroacetophenone (from Example C (57)) and 6-isothiocyanato-1H-benzoimidazole (see Boev et al., Pharm. Chem. J.) 24, (1990), 818-822) gave, after column chromatography with 1% HOAc / 10% MeOH / CH ₂ Cl ₂ as eluent, a 26% yield of a yellow powder, mp 250-252 ° C.

δ NMR (DMSO-d ₆ ): δ 8.18 (1H, s), 8.00 (2H, d, J = 8.7 Hz), 7.80 (1H, d, J = 8.7 Hz) 7.52 (1H, bd, J = 8.1 Hz), 7.24-7.10 (2H, m). IR (KBr): 3385, 1607, 1500 cm ^-1 .

HRFABMS: Calculated for C ₁₇ H ₉ Cl ₂ N ₆ O ₃ S (MH ⁺ ): 447.9930. Found:

447.9930.

Anal. Calcd for _{C 7} H ₁₀ Cl N ₂ O ₃ S _g. 0.1 H ₂ O. 1 MeOH. 0.7 HOAc. 0.1 CH ₂ C1 _2: C, 43.30; H, 3.35; N, 15.54; S, 5.93; Cl, 14.42. Found: C, 43.26; H, 3.01; N, 14.74; S, 7.14; Cl, 14.74.

♦ Example C (90):

/ 4-Amino-2- (4- (4-methylpiperazin-l-yl) -phenylamino / -thiazol-5-yl-1 / - (2, 5-dimethyl-thiophen-3-yl) -methanone H ₃ C <

The title compound was prepared in a manner similar to that in Example C (1). 1- (4-Isothiocyanato-phenyl) 4-methylpiperazine (from Example 0 (70)) and 3-bromoacetyl-2,5-dimethylthiophene (from Example C (50)) gave, after purification by flash column chromatography with 5-10% MeOH: CH ₂ Cl ₂ as a stepwise eluent, yellow solid in 70% yield, mp 205-206.5 ° C.

103 ¹ H NMR (DMSO-d ₆ ): δ 10.50 (1H, s), 8.00 (2H, bs), 7.48 (2H, d, J = 8.7 Hz), 6.95 (2H δ d, J = 8.7 Hz), 6.80 (1H, s), 3.10 (4H, dd, J = 5.0, 4.4 Hz), 2.46 (4H, t, J = 4.7 Hz), 2.42 (3H, s), 3.38 (3H, s), 2.24 (3H, s).

IR (KBr): 3154, 2944, 2804, 1609, 1543, 1516, 1420, 1296 ^cm-1 ·

HRFABMS. Calcd. for C ₂₁ H ₂₆ N ₅ OS ₂ (MH ⁺ ): 428.1579. Found: 428.1566

Anal. calculated for C ₂₁ H ₂₅ N ₅ OS ₂ . 0.7 H ₂ O: C, 57.30; H, 6.04; N, 15.91; S, 14.57. Found: C, 57.19; H, 6.06; N, 15.77; S, 14.55.

* Example C (91):

4-Amino-2- (1H-benzoimidazol-6-ylamino) -thiazol-5-yl-7- (3-amino-4-bromo-2,6-dichloro-phenyl) -methanone

3'-Amino-4'-bromo-2 ', 6'-dichloro-acetophenone having the structural formula

was prepared by the first following procedure. 3'-Amino-2 ', 6'-dichloroacetophenone (from Example C (82); 2.15 g, 11.3 mmol) in HOAc (8.7 mL) was carefully degassed with argon and cooled to 0 ° C; bromine was added and the reaction mixture was then allowed to warm to ambient temperature. After 0.5 h, the mixture was diluted with ice / water and extracted with ether. The combined ether layers were washed with saturated aqueous NaHCO ₃ and brine, dried over K ₂ CO ₃ and evaporated to give 2.87 g (90%) of a brown solid which was used without further purification.

3 '-Amino-2,4' structural formula

-dibromo-2 ', 6'-dichloroacetophenone having

r rt

104 was prepared in a manner analogous to 5-bromoacetyl-4-methyl-1H-imidazole of Example C (40).

3'-Amino-4'-bromo-2 ', 6'-dichloroacetophenone gave, after column chromatography with a step gradient of 2.5-5% CH ₂ Cl ₂ / hex, 725 mg (22% yield) of a white solid which was used without further purification. These fractions gave 33% recovery of the starting material.

The title compound was prepared in a manner analogous to that used in Example C (1). 6-Isothiocyanato-1H-benzoimidazole (see Boev et al., Pharm. Chem.

J. (English translation), vol. 24 (1990), pages 818-822) and 3'-amino-2,4'-dibromo-2 ', 6'-dichloroacetophenone gave pale yellow crystals in 34% yield, m.p. 227-230 [deg.] C.

1 H NMR (DMSO-d ₆ ): 512.48 (1H, bs), 10.85 (1H, s), 8.22 (1H, s), 8.06 (2H, bs), 7.80 (1H, bs) 7.34 (1H, s), 7.58 (1H, d, J = 8.5Hz), 7.22 (1H, d, J = 8.5Hz), 5.75 (2H, s).

FABMS (MH ⁺ ): Calcd .: 498.9333. Found: 498.9312.

Anal. calcd for C ₁₇ H _for N ₆ OSCl ₂ Br. 0.8 H ₂ O: C, 39.83; H, 2.48; N, 16.39; S, 6.26. Found: C, 39.92; H, 2.43; N, 16.26; S, 6.14.

* Example C (92):

4- [4-Amino-5- (2,5-dichlorothiophene-3-carbonyl) -thiazol-2-ylamino] -benzenesulfonamide

3-Bromoacetyl-2,5-dichlorothiophene, structural formula ⁸¹ was prepared in a manner similar to 2-bromo-2'-iodoacetophenone, see Example C (12). 3-Acetyl-2,5-dichlorothiophene (2.0 g, 10.2 mmol) rr ·

105 gave 2.9 g (100% yield) of a yellow oil which was used without further purification.

Δ, NMR (CDCl ₃ ): δ7.25 (1H, s), 4.40 (2H, s).

The title compound was prepared in a manner analogous to that used in Example C (1). 4-Isothiocyanato-benzenesulfonamide and 3-bromoacetyl-2,5-dichlorothiophene gave a yellow solid in 65% yield, m.p. Mp 274-276 ° C.

1 H NMR (DMSO-d ₆ ): δ 11.20 (1H, s), 8.24 (2H, bs), 7.80 (4H, s), 7.33 (1H, s), 7.31 ( 2H, s).

FABMS (MH ^&lt; ^{+ &gt;} ): 449/451.

Anal. calcd for C _u H ₁₀ N ₄ O ₃ S ₃ Cl _2: C, 37.42; H, 2.24; N, 12.47; S, 21.41, Cl, 15.78. Found: C, 37.56; H, 2.19; N, 12.39; S, 21.29; Cl. 1.71.

* Example C (93):

[4-Amino-2- (1H-benzoimidazol-6-ylamino) -thiazol-5-yl] - (2,5-dichloro-thiophen-3-yl) -methanone

The title compound was prepared in a manner analogous to that used in Example C (1). 6-Isothiocyanato-1H-benzoimidazole (see Boev et al., Pharm. Chem. J. (English translation), vol. 24 (1990), pp. 818-822) and 3-bromoacetyl-2,5- dichlorothiophene (from Example C (92)) gave, after precipitation of THF, an amorphous yellow solid in 52% yield, m.p. higher than 300 ° C.

Ή NMR (DMSO-d 6): δ 12.52 (1H, bs), 10.89 (IH, s), 8.26 (IH, s), 8.21 (2H, bs), 7.90 (IH (bs), 7.60 (1H, d, J = 8.4Hz), 7.28 (1H, s), 7.27 (IH, d, J = 8.4Hz).

Ř © r ©

Γ Γ

106 .

ESIMS (M ⁺ ): 410/412.

Anal. calculated for C ₁₅ H ₉ N ₅ OS ₂ Cl ₂ . 0.1 HCl. 0.6 THF: C, 45.71; H, 3.06; N, 15.32; S, 14.03, Cl, 16.28. Found: C, 45.84; H, 2.83 N, 15.01; S, 14.27; Cl, 16.00.

* Example C (94):

[4-Amino-2- [3- (4-methyl-piperazin-1-yl) -phenylamino] -thiazol-5-yl] - (3-methyl-5-nitro-thiophen-2-yl) -methanone

The title compound was prepared in a manner analogous to that used in Example C (1). 1- (4-Isothiocyanato-phenyl) -4-methyl-piperazine (from Example C (70)) and 2-bromoacetyl-3-methyl-5-nitro-thiophene (from Example C (78)) yielded, after precipitation with aqueous EtOH, amorphous dark brown solid in 64% yield.

¹ H NMR (DMSO-d ₆ ): 510.88 (1H, s), 8.38 (2H, bs), 8.04 (1H, s), 7.38 (2H, d, J = 9.0 Hz), 6.98 (2H, d, J = 9.0 Hz), 3.35 (4H, bs), 3.15 (4H, bs), 2.34 (3H, s), 2.28 ( 3H, s).

HRFABMS (MH ⁺ ): Calcd .: 459.1273. Found: 459.1259.

Anal. calculated for C ₂₀ H ₂₂ N ₆ O ₃ S ₂ . 0.8 H ₂ O. 0.2 EtOH: C, 50.18; H, 5.18; N, 17.43; S, 13.30. Found: C, 50.94; H, 4.98; N, 17.13; S, 13.55.

* Example C (95):

4- [4-Amino-5- (3-methyl-5-nitro-thiophene-2-carbonyl) -thiazole-2 H]. ylamino / benzenesulfonamide

no ₂ ·

107 ·. .

The title compound was prepared in a manner analogous to that used in Example C (1). 6-Isothiocyanato-benzenesulfonamide and 3-bromoacetyl-5-nitrothiophene (from Example C (78)) gave a dark brown solid in 38% yield, m.p. M.p. 268-269 ° C.

¹ H NMR (DMSO-d ₆ ): δ 11.31 (1H, s), 8.46 (2H, bs), 8.08 (1H, s), 7.81 (4H, s), 7.32 (2H, s), 2.38 (3H, s).

Anal. Calcd. for C ₁₅ H ₁₃ N ₅ O ₅ S ₃ : C, 40.99; H, 2.98; N, 15.94; S, 21.89. Found: C, 41.11; H, 2.95; N, 15.66; S, 21.70.

* Example C (96):

(3-Amino-4-bromo-2,6-dichlorophenyl) -4-amino-2- [4- (4-methylpiperazin-1-yl) -phenylamino] -thiazol-5-yl] -methanone

The title compound was prepared in a manner analogous to that used in Example C (1).

1- (4-Isothiocyanato-phenyl) -4-methyl-piperazine (from Example C (70)) and 3'-amino-2,4'-dibromo-2 ', 6'-dichloroacetophenone (from Example C (91) ) gave, after recrystallization from EtOH, a yellow powder in 43% yield, mp Mp 180-182 ° C.

¹ H NMR (DMSO-d 6): δ 10.61 (1H, s), 8.01 (2H, bs), 7.59 (1H, s), 7.28 (2H, d, J = 8.7) Hz), 6.94 (2H, d, J = 8.7 Hz), 5.74 (2H, s), 3.11 (4H, bs), 2.45 (4H, bs), 2.23 ( 3H, s).

HRFABMS (MH ⁺ ): Calcd .: 555.0136 / 557/559. Found: 555.0122 / 557/559.

Anal. calculated for C ₂₁ H ₂₁ N ₆ OSCl ₂ Br ₂ . 0.7 H ₂ 0. 0.6 EtOH: C, 44.70; H, 4.39; N, 14.09; S, 5.38. Found: C, 44.84; H, 4.18; N,

13.95; S, 5.27.

r κ p r

108 .

* Example C (97):

2- / 4- (1-Acetyl-piperazin-4-yl) -phenylamino / -4-amino-thiazol-6-yl- (2,6-dichloro-phenyl) -methanone

1-Acetyl-4- (4-nitro-phenyl) -piperazine having the structural formula

was prepared in a first manner analogous to N- (3-acetyl-2,4-dichlorophenyl) -acetamide of Example C (82). 1- (4-nitro-phenyl) -piperazine gave a yellow solid in 83% yield, which matched previously described material ^X H NMR (Katz et al., J. Amer. Chem. Soc., Vol. 111 (1989); 'p.7554 -7557).

1-Acetyl-4- (4-amino-phenyl) -piperazine, which has the structural formula

was prepared in a manner analogous to that used in Example C (82) for

4- (4-Methyl-piperazin-1-yl) -aniline. 1-Acetyl-4- (4-nitro-phenyl) -piperazine gave an off-white powder in 100% crude yield, which was used without further purification.

¹ H NMR (CDCl ₃ ): δ 6.85 (2H, d, J = 8.7 Hz), 6.98 (2H, d, J = 8.7)

Hz), 3.78 (2H, dd, J = 5.3, 5.0 Hz), 3.62 (2H, t, J = 5.3, 5.0

Hz), 3.62 (2H, dd, J = 5.3, 5.0 Hz), 2.98-3.10 (4H, m), 2.18 (3H, e < 0 &gt;

with). .

1-Acetyl-4- (4-isothiocyanato-phenyl) -piperazine having the structural formula

was prepared in a manner analogous to that used in Example C (41) for

1- (4-isothiocyanat-phenyl) -ΙΗ-imidazole. 1-Acetyl-4- (4-amino-phenyl) -piperazine gave a cream colored powder in 88% yield.

1 H NMR (CDCl ₃ ): δ 7.18 (2H, d, J = 9.0 Hz), 6.82 (2H, d, J = 9.0 Hz), 3.78 (2H, dd, J = 5, 1.35 Hz), 3.64 (2H, dd, J = 4.9, 5.3 Hz), 3.16-3.27 (4H, m), 2.10 (3H, s).

The title compound was prepared as described in Example C (1). 2-Bromo-2 ', 6'-dichloroacetophenone (from Example C (52)) and 1-acetyl-4- (4-isothiocyanatophenyl) piperazine afforded the crude product, which was precipitated from hexane to give a cream solid at 37 ° C. % yield, mp 265-267 [deg.] C.

1 H NMR (DMSO-d ₆ ): δ 10.60 (1H, bs), 8.02 (2H, bs), 7.50 (2H, d, J =

1.9 Hz), 7.42 (1H, m), 7.38 (2H, bs), 6.98 2H, d, J = 9.0 Hz), 3.60 (4H, s), 3, 20-3.10 (4H, m); 2.00 (3H, s).

IR (KBr): 3377, 3166, 1601, 1542, 11425 cm &^lt; -1 & gt ^; .

HRFABMS: Calcd. for C ₂₂ H ₂₂ Cl ₂ N ₅ O ₂ S (MH ⁺ ): 490.0871. Found: 490.0858.

Anal. calculated for C ₂₂ H ₂₁ Cl ₂ N ₅ O ₂ S. 0.16 H ₂ O. 0.1 C ₅ H ₁₄ : C, 54.08; H, 4.56; Cl, 14.13; N, 13.95; S, 6.39. Found: C, 53.88; H, 4.32; Cl, 14.46; N, 14.28; S, 6.54.

♦ Example C (98):

2- [4- (1-Acetyl-piperazin-4-yl) -phenylamino] -4-amino-thiazol-5-yl- (3-methyl-thiophen-2-yl) -methanone

110

The title compound was prepared in a manner analogous to that used in Example C (1). 1-Acetyl-4- (4-isothiocyanato-phenyl) -piperazine (from Example C (97)) and 2-bromoacetyl-3-methylthiophene (from Example C (19)) gave a yellow solid in 37% yield, m.p. Mp 290-292 ° C.

1 H NMR (DMSO-d ₆ ): 510.60 (1H, bs), 8.10 (2H, bs), 7.48 (1H, d, J = 5.0 Hz), 7.40 (2H, d, J = 8.7 Hz), 6.96-7.04 (2H, m), 3.60 (4H, s), 3.18 (2H, bs), 3.12 (2H, bs), 2.40 (3H, s); 2.02 (3H, s).

IR (KBr): 3377, 3166, 1633, 1601, 1542, 1425, 1225 cm ^-1 .

Anal. calculated for C ₂₁ H ₂₃ N ₅ O ₂ S. 1 H ₂ O: C, 56.89; H, 5.27; N, 15.80; S, 14.46. Found: C, 56.98; H, 5.27; N, 15.72; S, 14.35.

♦ Example C (99):

4- (4-Amino-5- (2-fluoro-6-trifluoromethyl-benzoyl) -thiazol-2-yl-amino) -benzenesulfonamide

2-Bromo-2'-fluoro-6'-trifluoromethylacetophenone having the structural formula was prepared in a manner analogous to

2-bromo-2'-iodoacetophenone, see Example C (12).

2'-Fluoro-β '- (trifluoromethyl) acetophenone (745 mg, 3.61 mmol)

Γ «*

111 gave 1.05 g of a yellow oil which was used without further purification.

The title compound was prepared in a manner analogous to that used in Example C (1). 4-Isothiocyanato-benzenesulfonamide and crude 2-bromo-2'-fluoro-6'-trifluoromethyl-acetophenone gave a pale yellow solid in 21% yield, m.p. Mp 290-292 ° C.

1 H NMR (DMSO-d ₆ ): 511.15 (1H, s), 8.20 (2H, bs), 7.83-7.68 (7H, m), 7.31 (2H, s).

Anal. Calcd. for C ₁₇ H ₁₂ N, O ₃ S ₂ F ₄ : C, 44.35; H, 2.63; N, 12.17; S, 13.93. Found: C, 44.42; H, 2.64; N, 12.13; S, 13.94.

♦ Example C (100):

The title compound was prepared in a manner analogous to that used in Example C (1).

1- (4-Isothiocyanato-phenyl) -4-methyl-piperazine (from Example 0 (70)) and 2-bromo-2'-fluoro-6'-trifluoromethyl-acetophenone (from Example C (99)) afforded the crude product which was recrystallized from EtOH to give a yellow powder in 74% yield, mp Mp 155-158 ° C.

1 H NMR (DMSO-d ₆ ): δ 10.62 (1H, s), 8.06 (2H, bs), 7.72-7.62 (3H, m), 7.10 (2H, d, J = 8.7 Hz), 6.93 (2H, d, J = 8.7 Hz), 3.11 (4H, bs), 2.45 (4H, bs), 2.22 (3H, s) .

HRFABMS (MH ⁺ ): Calcd .: 480, 1481. Found: 480.1486

112 '. .

Anal. calculated for C ₂₂ H ₂₁ N ₅ OSF ₄ . 1.0 EtOH: C, 54.84; H, 5.18; N, 13.33; S, 6.10. Found: C, 55.11; H, 5.11; N, 13.31; S, 6.00.

♦ Example C (101):

4-Amino-2- [4- (1-tert-butoxycarbonyl-piperazin-4-yl) -phenylamino] -thiazol-5-yl- (2,6-difluorophenyl) -methanone

1-tert-butoxycarbonyl-4- (4-nitrophenyl) -piperazine, having the structural formula:

^{N- 2} was prepared by the first procedure. To a suspension of 1- (4-nitro-phenylpiperazine (2.00 g, 9.65 mmol) in dioxane (30 mL) was added diisopropylethylamine (1.48 mL, 10.6 mmol) and di-tert-butyl dicarbonate (2). (10 g, 9.65 mmol) After 12 h, the mixture was poured into water (100 mL) and extracted with EtOAc (2 x 50 mL) .The combined organic layers were dried over MgSO ₄ , filtered and concentrated in vacuo to give a yellow solid, which was recrystallized from EtOAc / hexane to give 2.2 g of yellow needles, which material was used without further purification.

¹ H NMR (CDCl ₃ ): δ 8.20 (2H, d, J = 9.3 Hz), 6.82 (2H, d, J = 9.3 Hz), 3.58-3.64 (4H) m), 3.28-3.44 (4H, m), 1.54 (9H, s).

1- (4-Amino-phenyl) -4-tert-butoxycarbonyl-piperazine having the structural formula:

was prepared in a manner analogous to that of

4- (4-Methyl-piperazin-1-yl) -aniline for Example C (70). 1-tert-butoxycarbonyl-4- (nitro-phenyl) -piperazine gave a pale f

113 '. · Brown gel in 100% raw material yield which was used without further purification.

¹ H NMR (CDCl 3): δ 6.84 (2H, d, J = 8.7 Hz), 6.67 (2H, d, J = 8.8 Hz), 3.58 (4H, dd, J = 5.1, 5.0 Hz), 2.97 (4H, dd, J = 5.2, 4.8 Hz), 1.52 (9H, s).

1-tert-butoxycarbonyl-4- (4-isothiocyanato-phenyl) -piperazine, which has the structural formula:

was prepared in a manner analogous to that of

1- (4-isothiocyanat-phenyl) -1H-imidazole for Example C (41).

1- (4-Amino-phenyl) -tert-butoxycarbonyl-4-piperazine gave cream-colored needles in 87% yield, which were used without further purification.

¹ H NMR (CDCl ₃ ): δ 7.18 (2H, d, J = 9.0 Hz), 6.82 (2H, d, J = 9.0 Hz), 3.64 (4H, t, J = 5.3Hz), 3.24 (4H, t, J = 5.3Hz), 1.54 (9H, s). .

The title compound was prepared in a manner analogous to that used in Example C (1).

1- tert -butoxycarbonyl-4- (4-isothiocyanato-phenyl) -piperazine; and

2-bromo-2 ', 6'-difluoro-acetophenone (from Example C (79)) gave the crude product, which was recrystallized from EtOH to give a yellow solid in 67% yield, m.p. 140-143 ° C.

¹ H NMR (DMSO-d ₆ ): δ 10.67 (1H, s), 8.13 (2H, bs), 7.59-7.45 (1H, m), 7.35 (2H, d, J) = 9.0 Hz), 7.23-7.13 (2H, m), 6.96 (2H, d, J = 9.0 Hz), 3.46 (4H, bs), 3.07 ( 4H, bs), 1.43 (9H, s).

HRFABMS (MH ⁺ ): Calcd .: 516.1881. Found: 516.1900.

Anal. calculated for C ₂₅ H ₂₇ N ₅ O ₃ SF ₂ . 0.8 H ₂ O. 0.8 EtOH: C, 56.56; H, 5.57; N, 12.99; S, 5.95. Found: C, 56.34; H, 5.54; N, 12.89;

S, 5.83.

114 * Example C (102):

4- [4-Amino-5- (2,6-difluoro-benzoyl) -thiazol-2-ylamino] -benzamide

4-Isothiocyanato-benzamide having the structural formula: ² was prepared by the first procedure of McKee et al., J. Am.Chem. Soc., Vol. 48 (1946), pp. 2506-2507. To a solution of 4-aminobenzamide (5.00 g, 36.7 mmol) in water (60 mL) and 38% aqueous HCl (15 mL) was added thiophosgene (3.08 mL, 40.4 mmol). After about 30 minutes, the resulting white precipitate was filtered off, washed with water and dried under high vacuum to give 4.66 g (78% yield) of a white powder which was used without further purification.

¹ H NMR (DMSO-d 6): δ 8.08 (1H, bs), 7.94 (2H, d, J = 8.7 Hz), 7.53 (2H, d, J = 8.7 Hz) 7.51 (1H, bs).

The title compound was prepared in a manner analogous to that used in Example C (1). 4-Isothiocyanato-benzamide and 2-bromo-2 ', 6'-difluoro-acetophenone (from Example C (79)) gave a yellow solid in 26% yield, m.p. 297-298 ° C.

¹ H NMR (DMSO-d 6): δ 11.07 (1H, s), 8.22 (2H, bs), 7.91 (1H, s), 7.88 (2H, d, J = 8.7) Hz), 7.66 (2H, d, J = 8.7 Hz), 7.62-7.50 (1H, m), 7.31 (1H, s), 7.27-7.18 (2H , m).

Anal. Calcd. for C ₁₇ H ₁₂ N ₄ SF ₂ : C, 54.54; H, 3.23; N, 14.97; S, 8.57. Found: C, 54.27; H, 3.27; N, 14.68; S, 8.35.

* Example C (103):

tert-Butyl (/ 4- &gt; 4-amino-5- (2,6-difluoro-benzoyl) -thiazol-2-ylamine, phenyl / methyl-amino) -acetate \ ₀

A-i-vír

H

115 'target. Butyl E (methyl- (4-nitro-phenyl) -amino / acetate) having the structural formula,

was prepared by the first procedure. To a solution of sarcosine tert-butyl ester hydrochloride (2.0 g, 11 mmol) in DMSO (6 mL) was added 4-fluoro-nitrobenzene (1.6 g, 11 mmol) and triethylamine (3.4 mL, 24 mmol). . The resulting mixture was heated to 100 ° C for 12 hours. The resulting yellow suspension was allowed to cool, diluted with water (100 mL) and extracted with ether (2 x 100 mL). The combined organic extracts were dried over MgSO ₄ , filtered and concentrated in vacuo to give yellow needles, which were recrystallized from ether / hexane to give 2.0 g of yellow needles, which were used without further purification.

¹ H NMR (CDCl ₃ ): δ 8.18 (2H, d, J = 9.3 Hz), 6.62 (2H, d, J = 9.7 Hz), 4.08 (2H, s), 3.20 (3H, s); 1.42 (9H, s).

tert -Butyl [(4-amino-phenyl) -methyl-amino-2-acetate, which has the structural formula, was prepared in a manner analogous to 4- (4-methyl-piperazin-1-yl) -aniline for Example C (70). Butyl [methyl (4-nitro-phenyl) -amino] -acetate gave a red oil in 95% yield of the crude which was used without further purification.

1 H NMR (CDCl ₃ ): δ 6, 60-6.80 (4H, m), 4.08 (2H, s), 3.20 (2H, bs)

3.80 (2H, s), 2.82 (3H, s), 1.42 (9H, s).

target. Butyl [(4-isothiocyanato-phenyl) -methyl-amino] -acetate having the structural formula]

was prepared r e

116 'by a procedure analogous to 1- (4-isothiocyanato-phenyl) az-imidazole for Example C (41). tert -Butyl [(4-amino-phenyl) -methyl-amino] -acetate gave a pale brown solid in 98% yield, which was used without further purification.

¹ H NMR (CDCl 3): δ7.10 (2H, d, J = 9.1 Hz), 6.52 (2H, d, J = 9.1 Hz), 3.90 (2H, s), 2, 92 (3H, s); 1.30 (9H, s).

The title compound was prepared in a manner analogous to that used in Example C (1). tert -Butyl / (4-isothiocyanato-phenyl) -methyl-amino / acetate and 2-bromo-2 ', 6'-difluoro-acetophenone (from Example C (79)) gave a cream powder in 34% yield, m.p. 200.0-200.5 ° C.

1 H NMR (DMSO-d 6): δ 7.44-7.56 (1H, m), 7.10-7.30 (4H, m), 6.62 (2H, d, J = 9.0 Hz) 4.08 (2H, s), 2.95 (3H, s), 1.32 (9H, s).

IR (KBr): 3248, 3142, 2978, 1725, 1619, 1537, 1466, 1231 cm &^lt; -1 & gt ^; .

Anal. calcd for C ₂₃ H ₂₄ F ₂ N ₄ O ₃ S: C, 58.22; H, 5.10; N, 11.81; S, 6.76. Found: C, 58.27; H, 5.1; Cl, N, 11.53; S, 6.63.

* Example C (104)

4-Amino-2- [4- (1-tert-butoxycarbonyl-piperazin-4-yl) -phenylaminothiazol-5-yl- (3-methyl-thiophen-2-yl) -methanone

The title compound was prepared in a manner analogous to that used in Example C (1). tert -Butoxycarbonyl-4- (4-isothiocyanato-phenyl) -piperazine (from Example C (101)) and 2-bromoacetyl-3-methyl-thiophene (from Example C (91)) yielded, after recrystallization with EtOAc / hexane, 387 mg (52% yield) of a yellow solid, mp 175-176 [deg.] C.

¹ H NMR (CDCl ₃ ): δ 7.00-6.85 (4H, m), 3.62 (4H, dd, J = 5.3, 5.0 Hz), 3.18 (4H, dd, J = 5.3, 5.0 Hz), 2.48 (3H, s), 1.42 (9H, s).

f * f

117 '·

IR (KBr): 3260/2978, 1725, 1684, 1601, 1531, 1419, 1231 cm ^-1 .

Anal. calcd for C ₂₄ H ₂₉ N ₅ O ₃ S _2: C, 57.68; H, 5.85; N, 14.02; S, 12.83. Found: C, 57.74; H, 5.82; Cl, N, 13.95; S, 12.95. ♦ Example C (105):

4-Amino-2- [4- (1-tert-butoxycarbonyl-piperazin-4-yl) -phenylamino] -thiazol-5-yl- (2,6-dichloro-phenyl) -methanone

The title compound was prepared in a manner analogous to that used in Example C (1). tert -Butoxycarbonyl-4- (4-isothiocyanato-phenyl) -piperazine (from Example C (101)) and 2-bromo-2 ', 6'-dichloroacetophenone (from Example C (52)) afforded a crude product which was purified by flash column chromatography with MeOH: CH ₂ Cl ₂ (2.5: 97.5) as eluent and azeotroped with hexane to give a yellow solid in 90% yield, mp 165-167 ° C.

¹ H NMR (CDCl ₃ ): δ 7.22 (2H, d, J = 9.0 Hz), 6.92 (2H, d, J = 9.0 Hz), 3.60 (4H, m), 3.18 (4H, m); 1.42 (9H, s).

IR (KBr): 3401, 3271, 2966, 1689, 1607, 1542, 1460, 1255 cm &^lt; -1 & gt ^; .

HRFABMS: Calcd. for C ₂₅ H ₂₃ N ₅ O ₃ ClS: (MH ⁺ ): 548.1290. Found: 548.1270.

Anal. calculated for C ₂₅ H ₂₇ N ₅ O ₃ Cl ₂ S. 0.1 C ₆ H ₁₄ : C, 55.23; H, 5.07; N, 12.58; S, 5.76. Found: C, 55.34; H, 5.28; N, 12.29; Cl, 12.48; S, 5.58.

* Example C (106):

(3-Acetamido-2,6-dichlorophenyl) - (1-amino-2- [4-tert-butoxycarbonylpiperazin-1-yl) amino-thiazol-5-yl] -methanone

The title compound was prepared according to the procedure described in Example C (1). tert -Butoxycarbonyl-4- (4-isothiocyanato-phenyl) -piperazine (from Example C (101)) and N- (3-bromoacetyl-2,4-dichlorophenyl) -acetamide (from Example C (82)) gave pale yellow solid in 57% yield, mp Mp 248-150 ° C.

¹ H NMR (CDCl ₃ ): δ 7.20 (2H, d, J = 9.0 Hz), 6.92 (2H, d, J = 9.0 Hz), 3.54-3.66 (4H) , m), 3.12-3.22 (4H, m), 2.28 (3H, s), 1.42 (9H, s).

IR (KBr): 3377, 3271, 3177, 2978, 1672, 1548, 1437, 1290, 1231 cm ^-1 .

HRFABMS: Calcd. for C ₂₇ H ₃₁ Cl ₂ N ₆ O ₄ S: (MH ⁺ ): 605.1505. Found: 605.1528.

Anal. calculated for C ₂₇ H ₃₀ Cl ₂ N _g O ₄ S. 1.3 H ₂ O: C, 51.56; H, 5.22; N, 13.36; Cl, 11.27; S, 5.10. Found: C, 51.50; H, 5.18; Cl, 11.15; N, 13.19; S, 4.99.

* Example C (107):

4- [4-Amino-5- (2,4,6-trichloro-benzoyl) -thiazol-2-ylamino] -benzenesulfonamide

2,4,6-Trichloroacetophenone having the structural formula was prepared first as follows. It was a modified procedure from Reynolds et al., Org. Son. Coli., Vol. IV (1963), p.

708-710. The Mg rings (283 mg, 11.3 mmol) and EtOH (0.25 mL) were r-

119 'CC1 ₄ added. The subsequent reaction was quenched before addition of a solution of diethyl malonate (1.71 mL, 11.33 mmol) in EtOH (0.91 mL) at a rate to maintain the reaction. After 30 minutes, the mixture was refluxed to consume Mg for one hour, and then allowed to cool. The solid was suspended in ether (25 mL) and then the solution was carefully added

2,4,6-trichlorobenzoyl chloride (2.50 g, 10.3 mL) in ether (5 mL). After 3 days, a solution of H ₂ SO ₄ (0.6 mL) in water (10 mL) was carefully added to dissolve the solids and extracted with ether (2 x 10 mL). The extracts were dried over MgSO ₄ and evaporated to a dull oil which was taken up in HOAc (3 mL), H ₂ O (2 mL) and H ₂ SO ₄ (0.33 mL) and heated under reflux. After 7.5 hours, the mixture was allowed to cool overnight. The mixture was basified with 1N NaOH (35 mL) and extracted with ether (3 x 10 mL). The combined ether layers were dried with MgSO ₄ and evaporated to give 1.80 g (78%) of a white solid, which was used without further purification (previously described by Baker et al., J. Chem. Soc. (1941), p. 796-802).

2-Bromo-2 ', 4', 6'-trichloroacetophenone, having the structural formula q q, was prepared in a manner analogous to 2-bromo-2'-iodoacetophenone in Example C (12). Crude 2 ', 4', 6'-trichloroacetophenone gave 1.27 g (94%) of gold crystals which were used without further purification (previously described by Baker et al., J. Chem. Soc. (1941), p. 796). -802).

Ή NMR: δ 7.42 (2H, s), 4.42 (s, 2H).

The title compound was prepared essentially as described in Example C (1) except that an excess of potassium tert-butoxide (2.2 equivalents) was used. 4-Isothiocyanato-benzenesulfonamide and 2-bromo-2 ', 4', 6'-trichloroacetophenone gave a dark brown gum which was purified by column chromatography with 10% MeOH / CHCl ₃ and precipitated from MeOH / CHCl ₃ to give 96 mg ( 21%) amorphous, pale yellow solid.

Ρ P e «fí r

120 '· · ^X H NMR (CD ₃ OD): δ7,87 (4H, dd, J = 14.6, 9.0 Hz), 7.60 (2H, s).

IR (KBr): 3312, 1593, 1545, 1459, 1421, 1161 cm ^-1 .

ESIMS (MH ^&lt; ^{+ &gt;} ): 477/479/481. (M '): 475/477/479.

Anal. calcd for C ₁₆ H _U C1 ₃ N ₄ O ₃ S _2: C, 40.22; H, 2.32; N, 11.73; Cl, 22.26; S, 13.42. Found: C, 40.12; H, 2.34; N, 11.56; Cl,

22.41; S, 13.43.

* Example C (108):

4- [4-Amino-5- (2'-difluoro-benzoyl) -thiazol-2-ylamino] -N-methyl-benzenesulfonamide

4-Amino-N-methyl-benzenesulfonamide having the structural formula ^HsC '

7 ~ ^nh 2 hon = / was prepared first as follows. N-Methyl-4-nitro-benzenesulfonamide (2.58 g, 11.9 mmol; Khanna et al., J. Med. Chem., Vol. 40 (1997), pp. 1619-1633) and 10% Pd / C (250 mg) in MeOH (60 mL) was stirred under a nitrogen atmosphere for 2 hours and filtered. The filtrate was concentrated in vacuo to give 2.17 g (98% yield) of colorless crystalline flakes, which corresponded ^X H NMR given in the literature (Khanna et al., J. Med. Chem., Vol. 40 (1997) p. 1619-1633) and were used without further purification.

„No

4-Isothiocyanato-N-methyl-benzenesulfonamide having the structural formula was prepared in a manner analogous to the 4-isothiocyanato-benzamide of Example C (102). 4-Amino-N-methyl-benzenesulfonamide (2.17 g, 11.7 mmol) yielded 2.10 g (79% yield) of a white powder which was used

Γ Γ <r

121 ^X without further purification H NMR _(DMSO-d6): 8 7.83 (2H, d, J = 8.4 Hz), 7.65 (2H, d, J = 8.4 Hz), 7.61 (1H, q, J = 4.9Hz), 2.43 (3H, d, J = 4.9Hz).

The title compound was prepared essentially as described in Example C (1). 4-Isothiocyanato-N-methyl-benzenesulfonamide and 2-bromo-2 ', 6'-difluoroacetophenone (from Example C (79)) gave the crude product which was extracted with 10% i-PrOH / CHCl ₃ and purified by column chromatography with 5% MeOH / CHCl ₃ to give an amorphous yellow powder in 41% yield which decomposed above 200 ° C.

¹ H NMR (DMSO-d ₆ ): δ 11.23 (1H, s), 8.33 (2H, bs), 7.81 (2H, d, J =

8.5 Hz), 7.54 (2H, d, J = 8.5 Hz), 7.62-7.41 (1H, m), 7.39 (1H, q, J = 5.0 Hz) 7.23 (2H, t, J = 7.1Hz), 2.41 (3H, d, J = 5.0Hz).

HRFABMS (MH ⁺ ): Calcd .: 425.0554. Found: 425.0566.

Anal. calculated for C ₁₇ H ₁₄ N ₄ O ₃ S ₂ F ₂ . 0.5 CH ₃ OH: C, 47.72; H, 3.66; N, 12.72; S, 14.56. Found: C, 47.56; H, 3.52; N, 12.72; S, 14.77. * Example C (109):

4- / 4-amino-5- (2, 6-difluoro-benzoyl) -thiazol-2-ylamino _<? -N, N-dimethyl-benzenesulfonamide

4-Amino-N, N-dimethylbenzenesulfonamide having the structural formula κ ₃ ς fl

H ₃ C 6 • nh ₂ was prepared first as follows. Crude N, N-dimethyl-4-nitro-benzenesulfonamide (3.89 g,

16.9 mmol; Khanna et al., J. Med. Chem., Vol. 40 (1997) p.

1619-1633), 10% Pd / C (800 mg), MeOH (80 mL) and THF (200 mL) were r r

0

122 were stirred under an atmosphere of hydrogen for 6 hours and filtered. The filtrate was concentrated in vacuo to give 3.68 g of yellow solid, which corresponded to the spectrum ^X H-NMR described in Khanna et al., J. Copper. Chem., Vol. 40 (1997), pp. 1619-1633, and was used without further purification.

4-Isothiocyanato-N, N-dimethyl-benzenesulfonamide having the structural formula was prepared as follows. To a solution of 4-amino-N, N-dimethylbenzenesulfonamide (2.0 g, 10 mmol) in acetone (50 mL) at 5-10 ° C was added simultaneously a solution of thiophosgene (0.91 mL, 12 mmol) in acetone (20 mL). ml) and 25% aqueous Na ₂ CO ₃ (10 ml). After 5 minutes at 5-8 ° C, the mixture was allowed to warm and was stirred at room temperature for half an hour. The solvent was evaporated and water (70 ml) was added. The resulting light yellow precipitate was filtered off, washed with water and dried under vacuum to give 2.35 g (97% yield) of a white powder which was used without further purification.

Ή NMR (DMSO-d ₅ ): δ 7.82 (2H, d, J = 8.4 Hz), 7.69 (2H, d, J = 8.4 Hz), 2.63 (6H, s) .

The title compound was prepared essentially as described in Example C (1). 4-Isothiocyanato-N, N-dimethylbenzenesulfonamide and 2-bromo-2 ', 6'-difluoroacetophenone (from Example C (79)) gave a crude brown solid which was recrystallized from EtOH to give light brown crystals in 52% yield. , tt Mp 240-242 ° C.

^H NMR (DMSO-d _s): δ 11.24 (1H, s), 8.14 (2H, bs), 7.84 (2H, d, J = 8.8 Hz), 7.72 (2H δ d, J = 8.8 Hz), 7.62-7.49 (1H, m), 7.23 (1H, d, J = 7.9 Hz), 7.20 (1H, d, J = 8.0 Hz), 2.59 (6H, s).

Anal. Calcd. for C ₁₈ H ₁₆ N ₄ O ₃ S ₂ F ₂ : C, 49.31; H, 3.68; N, 12.78; S, 14.63. Found: C, 49.29; H, 3.71? N, 12.68; S, 14.50.

Γ -

r - r

123 * Example C (110):

(4-Amino-2- [4 - [(2-dimethylamino-ethyl) -methyl-amino] -phenylamino] -thiazol-5-yl) - (2,6-difluoro-phenyl) -methanone

N- (4-Nitrophenyl) -Ν, N structural formula, N'-trimethyl-ethane-1,2-diamine having

. was prepared in a first manner analogous to tert-butyl / methyl (4-nitro-phenyl) -amino] -4-acetate from Example C (103)). 4-Fluoronitrobenzene and Ν, Ν, Ν'-trimethylethylenediamine gave a brown oil in 87% yield of the raw material, which were used without further purification.

1 H NMR (CDCl ₃ ): δ 8.14 (2H, d, J = 9.6 Hz), 6.64 (2H, d, J = 9.3)

Hz), 3.58 (2H, t, J = 7.5 Hz), 3.12 (3H, s), 2.52 (2H, t, J = 7.5 Hz), 2.32 (6H, with).

N- (4-Aminophenyl) -N, N ', N'-ethane-1,2-diamine having the structural formula ch ₃ was prepared in a manner analogous to 4- (4-methyl-piperazin-1-yl) -aniline from Example C (70). N- (4-nitrophenyl) - Ν, Ν, Ν'-trimethyl-ethane-1,2-diamine gave a reddish-brown oil in 92% yield of the raw material which was used without further purification.

¹ H NMR (CDCl ₃ ): δ 6.62 (4H, s), 3.30 (2H, dd, J = 7.6, 7.4 Hz), 2.85 (3H, s), 2.47 (2H, dd, J = 7.7, 7.2 Hz), 2.32 (6H, s).

N- (4-Isothiocyanato-phenyl) -N, N ', N'-trimetyl-ethyl-1,2-diamine,

124

having the structural formula ^CH 'was prepared in a manner analogous to 1- (4-isothiocyanato-phenyl) -1H-imidazole of Example No. (41). N- (4-Aminophenyl) -N, N ', N'-ethane-1,2-diamine gave a brown oil in 75% yield of the crude which was used without further purification.

¹ H NMR (CDCl ₃ ): δ 7.13 (2H, d, J = 8.8 Hz), 7.01 (2H, d, J = 8.2 Hz), 3.99 (2H, dd, J = 7.6, 7.1 Hz), 3.15 (1H, bs), 3.02 (3H, s), 2.80 (6H, s).

The title compound was prepared essentially as described in Example C (1). N- (4-Isothiocyanato-phenyl) -N, N ', N'-trimethyl-ethane-1,2-diamine and 2-bromo-2', 6'-difluoroacetophenone (from Example C (79)) gave the crude product which was purified by MeOH: CH ₂ Cl ₂ step gradient column chromatography (2.5: 97.5-10: 90) to give a yellow solid in 55% yield, mp: 96-98 ° C.

¹ H NMR (DMSO-d ₆ ): δ 7.42-7.55 (1H, m), 7.10-7.24 (4H, m), 6.64 (2H, d, J = 9.0 Hz), 2.90 (3H, s), 2.38 (2H, dd, J = 7.2, 6.5 Hz), 2.18 (6H, s).

IR (KBr): 3394, 3180, 2948, 2828, 1620, 1546, 1523, 1466 ^cm-first

HRFABMS: Calcd. for C ₂₁ H ₂₄ F ₂ N ₅ OS (MH +): 432, 1670. Found: 432.1658.

Anal. calculated for C ₂₁ H ₂₃ F ₂ N ₅ OS. 0.4 H ₂ O: C, 57.49; H, 5.47; N, 15.96; S, 7.31. Found: C, 57.36; H, 5.45; N, 15.77; S, 7.27.

* Example C (III):

2- / 4- (1-Acetyl-piperazin-4-yl) -phenylamino / -4-amino-thiazol-5-yl- (2,6-difluorophenyl) -methanone

r r λ.-γ γ r r f · Γ <'

125 '

The title compound was prepared essentially as described in Example C (1).

1-Acetyl-4- (4-isothiocyanato-phenyl) -piperazine (from Example C (97)) and 2-bromo-2 ', 6'-difluoroacetophenone (from Example C (79)) gave 320 mg (66% yield) ) cream colored solids, m 298 ° C.

¹ H NMR (DMSO-d ₆ ): δ 7.44-7.58 (1H, m), 7.36 (2H, bd, J = 7.2 Hz), 7.18 (2H, dd, J = 8.1, 7.5 Hz), 6.95 (2H, d, J = 9.0 Hz), 3.58 (4H, bs), 3.00-3.20 (4H, m), 2, 05 (3 H, s).

IR (KBr): 3389, 3154, 1607, 1601, 1542, 1419, 1231 cm &^lt; -1 & gt ^; .

HRFABMS: Calcd. for C ₂₂ H ₂₁ F ₂ N ₃ UNa (M + Na): 480.1282. Found: 480.1266.

Anal. calculated for C ₂₂ H ₂₁ N ₅ O ₂ F ₂ S. 0.3 H ₂ O: C, 57.08; H, 4.70; N, 15.31; S, 6.93. Found: C, 56.95; H, 4.74; N, 15.16; S, 6.82.

* Example C (112):

2- [4- (1-Acetyl-piperazin-4-yl) -phenylamino] -4-aruino-thiazol-5-yl- (2,5-dimethyl-thiophen-3-yl) -methanone

The title compound was prepared as described in Example C (1). 1-Acetyl-4- (4-isothiocyanato-phenyl) piperazine (from Example C (97)) and 3-bromoacetyl-2,5-dimethylthiophene (from Example C (50)) yielded 200 mg (53% yield) of a pale cream colored solid, m.p. Mp 282-283 ° C.

Ή NMR (DMSO-d 6): δ 7.42 (2H, d, J = 9.0 Hz), 6.98 (2H, d, J = 9.0 Hz), 6.82 (1H, s), 3.60 (4H, bs), 3.02-3.20 (4H, m), 2.46 (3H, s), 2.38 (3H, s), 2.05 (3H, s).

IR (KBr): 3401, 3166, 1637, 1601, 1542, 1425, 1231 cm &^lt; -1 & gt ^; .

126

HRFABMS: Calcd. for C ₂₂ H ₂₆ N ₅ O ₂ S ₂ (MH ⁺ ): 456.1528. Found: 456.1510.

Anal. Calcd. for C ₂₂ H ₂₅ N ₅ O ₂ S: C, 57.87; H, 5.74; N, 15.34; S, 14.05. Found: C, 57.85; H, 5.53; N, 15.23; S, 14.20.

* Example C (113):

4- [4-Amino-5- (2-fluoro-6-trifluoromethyl-benzoyl) -thiazol-2-ylamino] -benzamide

The title compound was prepared in a manner analogous to that of Example C (1). 4-Isothiocyanato-benzamide (from Example C (102)) and 2-bromo-2'-fluoro-6'-trifluoromethylacetophenone (from Example C (99)) gave the crude product, which was purified by step gradient column chromatography

8-10% EtOH / CHCl ₃ to give an amorphous yellow solid in 14% yield which decomposed above 145 ° C.

Ή NMR (DMSO-d ₆ ): δ 8.30 (1H, bs), 8.10 (IH, bs), 7.94-7.82 (3H, m), 7.74-7.62 (5H) , m), 7.30 (1H, s).

HRFABMS (MHU: Calcd .: 425.0695. Found: 425.070.

Anal. calculated for C ₁₈ H ₁₂ N ₄ O ₂ SF ₄ . 0.9 EtOH: C, 51.05; H, 3.76; N, 12.03; S, 6.88. Found: C, 51.14; H, 3.78; N, 1236; S, 6.79.

* Example C (114):

4- ₍ 4-Amino-5- (3-methyl-thiophene-2-carbonyl) thiazol-2-ylamino) -N-methylbenzenesulfonamide

r e e e r

127 '·

The title compound was prepared in a manner analogous to that used in Example C (1). 4-Isothiocyanato-N-methyl-benzenesulfonamide (from Example C (108)) and 2-bromoacetyl-3-methylthiophene (from Example C (19)) gave a yellow solid in 57% yield, m.p. 197.0-199.5 ° C.

¹ H NMR (DMSO-d 6): δ 11.19 (1H, s), 8.24 (2H, bs), 7.86 (2H, d, J = 8.7 Hz), 7.75 (2H, d, J = 8.7 Hz), 7.65 (1H, d, J = 5.0 Hz), 7.36 (1H, q, J = 6.1 Hz), 7.03 (1H, d, J = 5.0 Hz), 2.42 (3H, s), 2.41 (3H, d, J = 6.1 Hz).

HRFABMS (MH ⁺ ): Calcd .: 409.0463. Found: 409.0474.

Anal. calcd for C ₁₆ H ₁₆ N ₄ O ₃ S _3: C, 46.23; H, 4.07; N, 13.48; S, 23.14. Found: C, 46.28; H, 3.98; N, 13.38; S, 23.08.

* Example C (115)

4- [Amino-5- (2,4, β-trifluorobenzoyl) -thiazol-2-yl-amino] -benzenesulfonamide

2-Chloro-2 ', 4', 6'-trifluoroacetophenone having ci.

structural formula JL 1

I was prepared subsequently. To a mechanically stirred solution of 1,3,5-trifluorobenzene (5.17 mL, 50.0 mmol) in dichloromethane (12.5 mL) was gradually added AlCl ₃ (13.4 g, 115 mmol) gradually over 15 minutes. A sharp gargle and the development of HCl gas were observed. The mixture was carefully heated under reflux and chloroacetyl chloride (6.20 g, 4.37 mL, 55.0 mmol) was added dropwise over 45 minutes. After 6 hours under reflux, the mixture was allowed to cool for 12 hours, then was carefully poured onto ice / water slurry (about 200 mL) and extracted with ether (3 x 50 mL). The combined ether layers were washed with 10% aqueous HCl (2 x 30 mL),

C ř

128

1N aqueous NaOH (3 x 30 mL), and brine (25 mL), dried over MgSO ₄ and evaporated to give 5.28 g (51%) of a yellow solid which was used without further purification. An analytical sample was crystallized from ether / hexane to give yellow microcrystals, mp 43-45 ° C.

¹ H NMR (CDCl 3): δ 6.81- (2H, t, J = 8.4 Hz), 4.54 (2H, s).

IR (KBr) 1721, 1637, 1616, 1447, 1201, 1128, 1045 cm &^lt; -1 & gt ^; .

Anal. Calcd. for C ₈ H ₄ ClF ₃ O: C, 46.07; H, 1.93; Cl, 17.00. Found: C, 45.92; H, 1.95; Cl, 16.97.

The title compound was prepared essentially in a manner analogous to that described in Example C (1) except that an excess of potassium tert-butoxide (2.2 equivalents) was used. 4-Isothiocyanato-benzenesulfonamide and 2-chloro-2 ', 4', 6'-trifluoroacetophenone gave a red-brown solid, which was purified by column chromatography with 5% MeOH / CH ₂ Cl ₂ as eluent. Precipitation with traces of hexane in MeOH / CH ₂ Cl ₂ gave 70 mg (33%) of a yellow amorphous powder which decomposed above 148 ° C.

¹ H NMR (CD ₃ OD): δ 7.91 (1H, s), 7.86 (4H, dd, J = 14.9, 6.9 Hz), 6.99 (2H, dd, J = 9) (0.5 Hz).

IR (KBr): 3278, 1602, 1549, 1425, 1155 ^cm-first

HRFABMS: Calcd. for C ₁₆ H ₁₂ F ₃ N ₄ O ₃ S ₂ : (MH ⁺ ): 429.0303. Found: 429.0315.

Anal. calcd for C ₁₆ H _U F ₃ N ₄ O ₃ S _2nd 1.1 H ₂ 0 :, C, 42.87; H, 2.79; N, 12.50; S, 14.31. Found: C, 42.98; H, 2.73; N, 12.12; S, 14.48; * Example C (116) [4-Amino-2 - [(4-methyl-piperazine-1-sulfonyl) -phenylamino] -thiazol-5-yl] - (2,6-difluorophenyl) -methanone </RTI> 000 0 _r 0 0, -rf C 0 0 1 0 r 0 y

129 · 1-Methyl-4- (4-nitro-benzenesulfonyl) -piperazine, which has the structural formula _y . .__ of η ₃ ο - ^^ _{/ Ν} - ^^ _ _Λ κ _ΝΟ2 was prepared in a manner analogous to that used for N-methyl-4-nitro-benzenesulfonamide for Example C (108) (Khanna et al., J. Med Chem, vol 40 (1997), pp. 1619-1633). 4-Nitrobenzenesulfonyl chloride and 1-methylpiperazine gave 5.1 g (88% yield) of a yellow solid which was used without further purification.

4- (4-Methyl-piperazin-l-sulfonyl) -aniline, which has the structural formula _ and _ was prepared ^H3C-N ^ ₂ _nh manner analogous to that used for

N-methyl-4-aminobenzenesulfonamide from Example C (108).

1-Methyl-4- (4-nitrobenzenesulfonyl) piperazine gave a gray solid in 99% yield, which was used in the next step without further purification.

¹ H NMR (DMSO-d ₆ ): δ7.37 (2Η, d, J = 8.8 Hz), 6.67 (2H, d, J = 8.8 Hz), 6.16 (2H, bs) 3.30 (4H, bs), 3.03 (4H, bs), 2.58 (3H, s).

1- (4-Isothiocyanato-benzenesulfonyl) -methyl-piperazine having the structural formula I-9 was prepared in a manner analogous to 4-isothiocyanato-benzamide in Example C (102). 4- (4-Methyl-piperazine-1-sulfonyl) -aniline yielded 1.1 g. (94%) yield of white crystals which were used without further purification.

¹ H NMR (CDCl ₃ ): δ 7.74 (2H, d, J = 8.6 Hz), 7.35 (2H, d, J = 8.6 Hz), 3.27 (4H, bs), 2.77 (4H, bs), 2.47 (3H, s).

The title compound was prepared essentially by a procedure analogous to that described in Example C (1).

r e r r

130 ·

1- (4-Isothiocyanato-benzenesulfonyl) -4-methyl-piperazine a

2-bromo-2 ', 6'-difluoroacetophenone (from Example C (79)) gave a yellow solid in 69% yield, m.p. M.p. 172-174 ° C.

¹ H NMR (DMSO-d ₆ ): δ 11.23 (1H, bs), 8.21 (2H, bs), 7.84 (2H, d, J = 8.8 Hz), 7.69 (2H , d, J = 8.8 Hz), 7.62-7.49 (1H, m). 7.22 (1H, d, J = 7.8Hz), 7.19 (IH, d, J = 8.1Hz), 2.87 (4H, t, J = 4.5Hz), 2, 35 (4H, t, J = 4.5Hz), 2.13 (3H, s).

HRFABMS (MH ⁺ ): Calcd .: 494.1132. Found: 494.1120.

Anal. calcd for C ₂₁ H ₂ IN ₃ O ₃ S ₂ F _2nd 0.1 H ₂ O. 0.5 CH ₃ OH: C, 50.50; H, 4.57; N, 13.70; S, 12.54. Found: C, 50.34; H, 4.39; N, 13.51; S, 12.63.

* Example C (117) (4-Amino-2- / 4 - [(2-dimethylamino-ethyl) -methyl-amino] -phenylamino] -thiazol-5-yl) - (3-methyl-thiophen-2-yl) ) -methanone

The title compound was prepared as described in Example C (1). N- (4-Isothiocyanato-phenyl) -N, N ', N'trimethylethane-1,2-diamine (from Example C (110) and 2-bromoacetyl-3-methylthiophene (from Example C (19) yielded, after purification) flash column chromatography with MeOH: CH ₂ Cl ₂ (5:95) as eluant, a yellow foam in 70% yield.

1 H NMR (DMSO-d 6): δ 7.22 (1H, d, J = 5.0 Hz), 7.16 (2H, d, J = 9.0 Hz), 6.72 (1H, d, J = 5.0 Hz), 6.58 (2H, d, J = 9.0 Hz), 3.44 (2H, dd, J = 7.7, 7.4 Hz), 3.00 (3H, s ), 2.42 (3H, s), 2.3 (6H, s).

IR (KBr): 3377, 3269, 2937, 2821, 1609, 1543, 1518, 1423 ^cm-first

HRFABMS: Calcd. For C ₂₀ H ₂₆ Cl ₂ N ₅ OS ₂ (MH ⁺ ): 416.1579. Found: 416.1594.

yy yy yy yy yy yy yy yy yy yy yy yy yy yyy c r

131 ·

Anal. calculated for C ₂₀ H ₂₅ Cl ₂ N ₅ OS ₂ . 1 H ₂ O: C, 55.40; H, 6.28; N, 16.15; S, 14.71. Found: C, 55.43; H, 5.94; N, 16.37; S, 14.57.

* Example C (118)

4- (4- / 4-Amino-5- (2,6-difluorobenzoyl) -thiazol-2-ylamino) -phenyl] -1-methyl-piperazin-2-one

4- (4-Nitro-phenyl) -piperazin-2-one having the structural formula v

^h v / o-no ₂ was produced by a first method analogous to tert-butyl / methyl (4-nitrophenyl) amino) acetate in Example C (103). Piperazin-2-one (Aspinall et al., J. Amer. Chem. Soc., Vol. 62 (1940), pp. 1202-1204) and 4-fluoronitrobenzene gave a yellow solid in 63% yield, which was used without any further purification.

¹ H NMR (CDCl ₃ ): δ 8.10 (2H, d, J = 8.8 Hz), 6.80 (2H, d, J = 9.2 Hz), 6.38 (1H, bs), 4.10 (2H, s), 3.74-2.52 (4H, m).

1-Methyl-4- (4-nitrophenyl) -piperazin-2-one, the formulas were further prepared as follows.

h ₃ cn no ₂

To a suspension of 4- (4-nitro-phenyl) -piperazin-2-one (500 mg, 2.26 mmol) in THF (5 mL) was added NaH (60 mg, 2.5 mmol). The mixture was cooled to 0 ° C, iodomethane (162 µΐ, 2.59 mmol) was added, and then allowed to warm to ambient temperature. After 12 hours, the solvent was removed in vacuo to give a yellow resin which was treated with H ₂ O. The yellow precipitate obtained was filtered, washed with H ₂ O and dried under high vacuum for several hours to give 420 mg (79% yield).

¹ H NMR (CDCl 3): δ 8.18 (2H, d, J = 9.4 Hz), 6.78 (2H, d, J = 9.4 Hz), 4.08 (2H, s), 3.68 (2H, dd, J = 4.7, 3.6 Hz), 3.54 (2H, dd,

132

J = 4.9, 3.7 Hz), 3.02 (3H, s).

4- (4-Aminophenyl) -1-methyl-piperazin-2-one having the structural formula

was prepared in a manner analogous to 4- (4-methyl-piperazin-1-yl) -aniline of Example C (70). 1-Methyl-4- (4-nitro-phenyl) -piperazin-2-one gave a brown resin which was used without any further purification.

¹ H NMR (CDCl ₃ ): δ 6.78 (2H, d, J = 9.0 Hz), 6.60 (2H, d, J = 9.0 Hz), 3.76 (2H, s), 3.44 (2H, dd, J = 5.8, 4.9 Hz), 3.20 (2H, dd,

J = 4.9, 4.0 Hz), 3.02 (3H, s).

4- (4-Isothiocyanato-phenyl) -1-methyl-piperazin-2-one having the structural formula

Was prepared in a manner analogous to 1- (4-isothiocyanato-phenyl) -1H-imidazole of Example C (41). 4- (4-Amino-phenyl) -1-methyl-piperazin-2-one gave a cream colored powder in 85% yield, which was used without further purification.

¹ H NMR (CDCl 3): δ 7.18 (2H, d, J = 9.0 Hz), 6.80 (2H, d, J = 9.0 Hz), 3.90 (2H, s), 3 50 (4H, bs), 3.70 (3H, s).

The title compound was prepared essentially by a procedure analogous to that described in Example C (1).

4- (4-Isothiocyanato-phenyl) -methyl-piperazin-2-one and 2-bromo-2 ', 6'-difluoroacetophenone (from Example C (79)) gave a yellow solid in 77% yield, m.p. greater than 300 ° C.

¹ H NMR (DMSO-d ₆ ): δ 7.60-7.70 (1H, m), 7.48 (2H, bd, J = 8.3 Hz), 7.31 (2H, t, J = 7.9 Hz), 7.09 (2H, d, J = 9.0 Hz), 3.88 (2H, s), 3.58 (4H, bd, J = 4.4 Hz), 3.02 (3 H, s).

133 '

Anal. Calcd for C ₂₁ H ₁₉ F ₂ N ₅ O ₂ S: C, 56.88; H, 4.32; N, 15.79; S, 7.23. Found: C, 56.81; H, 4.42; N, 15.83; S, 7.31.

* Example C (119) [4-Amino-2- (4-thiomorpholin-4-yl-phenylamino) -thiazol-5-yl] - (2,6-difluoro-phenyl) -methanone

4-Thiomorpholin-4-yl-aniline having the structural formula was prepared first subsequently. 4- (4-Nitro-phenyl) -thiomorpholine (1.50 g, 6.70 mmol; Beach et al., J. Chem. Soc. Perkin Trans. 2 (1984), pp. 217-221) and 10 % Pd / C (200 mg wet DeGussa type,

50%) were stirred in ethyl acetate (20 mL) and MeOH (20 mL) under hydrogen overnight and filtered. The filtrate was concentrated in vacuo to give 1.28 g (98% yield) of white crystalline flakes which were used without further purification.

4- (4-Isothiocyanato-phenyl) -thiomorpholine having the structural formula

was prepared in a manner analogous to that of

4-isothiocyanato-N, N-dimethylbenzenesulfonamide in Example C (109). 4-Thiomorpholin-4-yl-aniline gave a yellow powder in 83% yield.

1 H NMR (CDCl 3): δ 7.13 (2H, d, J = 9.1 Hz), 6.79 (2H, d, J = 9.1 Hz, 3.59 (4H, ddd, J = 5) 2, 5.0, 2.6 Hz), 2.72 (4H, ddd, J = 5.2, 5.2, 2.6 Hz).

The title compound was prepared essentially by a procedure analogous to that described in Example C (1).

4- (4-isothiocyanato-phenyl) thiomorpholine and 2-bromo-2 ', 6'-difluoroacetophenone (from Example C (79)) provided a yellow powder in 51% _r rr r rrrfrr' f Γ

134 yield; m.p. 128-130 [deg.] C.

¹ H NMR (DMSO-d ₆ ): δ 10.64 (1H, s), 8.12 (2H, bs), 7.56-7.44 (1H, m), 7.30 (2H, d, J = 9.0 Hz), 7.18 (1H, d, J = 7.7 Hz), 7.15 (1H, d, J = 8.1 Hz), 6.91 (2H, d, J = 9.0 Hz), 3.47 (2H, dd, J = 5.1, 5.0 Hz), 2.65 (2H, dd, J = 5.1, 5.0 Hz).

HRFABMS (MH ⁺ ): Calcd .: 433.0968. Found: 433.0980.

Anal. calc. for C ₂₀ H ₁₈ N ₄ OS ₂ F ₂ . 0.2 H ₂ O: C, 55.08; H, 4.25; N, 12.85; S, 14.71. Found: C, 55.02; H, 4.14; N, 12.72; S, 14.53. * Example C (120)

4- / 4- / 4-Amino-5- (1,6-difluorobenzoyl) -thiazol-2-ylaro-no-phenyl / -1-piperazin-2-one

4- (4-Amino-phenyl) -piperazin-2-one having the structural formula

It was made analogous to 4- (4-methyl-piperazin-1-yl) aniline (70) _from ^N - (2-y-NH ₂ ). 4- (4-Nitrophenyl) piperazin-2-one (from the example a pale brown oil in 100% yield of raw material which any further purification).

as in Example C (115)) was used without

1 H NMR (CD ₃ OD): δ 7.02 (2H, d, J = 8.7 Hz), 6.91 (2H, d, J = 8.8 Hz), 3.81 (2H, s), 3.59 (2H, dd, J = 5.9, 4.8 Hz), 3.46 (2H, dd, J = 5.9, 4.8 Hz).

4- (4-Isothiocyanato-phenyl) -piperazin-2-one having the structural formula _n 'o was prepared in a manner analogous to 1- (4-isothiocyanato-phenyl) -ΙΗ-imidazole in Example C (41). 4- (4-Amino-phenyl) -piperazin-2-one gave a cream

135 a colored solid that was used without further purification.

¹ H NMR (CDCl ₃ ): δ 9.00 (1H, bs), 8.20 (2H, d, J = 9.0 Hz), 7.80 (2H, d, J = 9.0 Hz), 4.50 (2H, s); 4.00-4.30 (4H, m). (3 H, s).

The title compound was prepared essentially by a procedure analogous to that described in Example C (1).

4- (4-Isothiocyanato-phenyl) -piperazin-2-one and 2-bromo-2 ', 6'-difluoroacetophenone (from Example C (79)) gave a yellow solid in 56% yield, m.p. Mp 280-282 ° C.

^X H NMR (DMSO-d _s): δ 9.12 (3H, s), 8.32-8.44 (IH, m), 8.18 (2H, bd, J = 6.9Hz), 8 0.05 (2H, t, J = 8.2 Hz), 7.78 (2H, d, J = 9.0 Hz), 4.52 (2H, s).

HRFABMS: Calcd. for C ₂₀ H ₁₈ F ₂ N ₅ O ₂ S (MHU: 430.1149. Found: 430.1138).

Anal. calculated for C ₂₀ H ₁₇ F ₂ N ₅ O ₂ S. 0.3 H ₂ O: C, 55.24; H, 4.08; N, 16.11; S, 7.37. Found: C, 55.24; H, 4.10; N, 15.77; S, 7.34.

* Example C (121):

[4-Amino-2- [4- (4-cyclopropylmethyl-piperazin-1-yl) -phenylamino] -thiazol-2-yl] - (2,6-difluoro-phenyl) -methanone

1-Cyclopropylmethyl-4- (4-nitro-phenyl) -piperazine, which has the structural formula > To a suspension of 1- (4-nitro-phenyl) -piperazine (2.50 g, 12.1 mmol) in DMF (10 mL) was added anhydrous Na ₂ CO ₃ (639 mg, 6.03 mmol) and bromomethylcyclopropane (585). μΐ, 6.03 mmol). The mixture was heated to 100 ° C overnight, then allowed to cool and diluted with water (30 mL). The separated aqueous layer was extracted with CHCl ₃ (3x

136 ml). The combined organic layers were dried over Na ₂ SO ₄ , filtered, and concentrated under reduced pressure to give an orange-brown solid which was purified by flash chromatography on a column with 2.5% MeOH / CH ₂ Cl ₂ as eluent to give 2.65 g (84% yield) of a yellow solid. This material was used without any further purification.

1 H NMR (CDCl ₃ ): δ 8.10 (2H, d, J = 10.7 Hz), 7.11 (2H, d, J = 9.5 Hz), 3.45 (4H, dd, J = 5.3, 5.1 Hz), 2.65 (4H, dd, J = 5.3, 5.1 Hz), 2.29 (2H, d, J = 6.6 Hz), 0.84 -0.98 (1H, m), 0.50-0.58 (2H, m), 0.10-0.15 (2H, m).

4- (4-Cyclopropylmethyl-piperazin-1-yl) -aniline having the structural formula was prepared in a manner analogous to 4- (4-methyl-piperazin-1-yl) -aniline in Example C (70). 1-Cyclopropylmethyl-4- (4-nitro-phenyl) -piperazine gave a red solid in 99% yield of the crude which was used without further purification.

1 H NMR (CDCl 3): δ 6.85 (2H, d, J = 9.9 Hz), 6.62 (2H, d, J = 8.8 Hz), 3.42 (2H, bs), 3 10 (4H, dd, J = 5.1, 4.8 Hz), 2.69 (4H, dd J = 5.1, 4.9 Hz), 2.30 (2H, d, J = 6, 5 Hz), 0.90-0.98 (1H, m),

0.50-0.56 (2 H, m), 0.10-0.15 (2 H, m).

1-Cyclopropylmethyl-4- (4-isothiocyanato-phenyl) -piperazine having the structural formula

was prepared in a manner analogous to 1- (4-isothiocyanat-phenyl) -ΙΗ-imidazole in Example C (41). 4- (4-Cyclopropylmethyl-piperazin-1-yl) -aniline gave a dark brown oil in 95% yield, which was used without further purification.

¹ H NMR (CDCl ₃ ): δ 6.80 (2H, d, J = 9.0 Hz), 6.68 (2H, d, J = 9.1 yyyy)

137 '

Hz), 3.08 (4H, bs), 2.55 (4H, bs), 2.10 (2H, d, J = 6.2 Hz), 0.65-0.80 (1H, m), 0.42 (2H, d, J = 8.0Hz), 0.00 (2H, d, J = 4.6Hz).

The title compound was prepared as described in Example C (1).

1- Cyclopropylmethyl-4- (4-isothiocyanato-phenyl) -piperazine a

2-bromo-2 ', 6'-difluoroacetophenone (from Example C (79)) gave, after crystallization from EtOH, a yellow solid in 17% yield, m.p. Mp 215-218 ° C.

¹ H NMR (DMSO-d 6): δ 10.60 (1H, s), 8.04 (2H, bs), 7.46-7.56 (1H, m), 7.18-7.20 (2H, m), 7.08 (2H, dd, J = 8.0, 7.7 Hz), 6.82 (2H, d, J = 9.1 Hz), 2.98-3.03 (4H, m 1.47 (4H, bs), 2.12 (2H, d, J = 6.6 Hz), 0.72-0.78 (1H, m), 0.34-0.42 (2H, m), 0.00-0.12 (12H, m).

IR (KBr): 2917, 1620, 1513, 1428 ^cm-first

HRFABMS: Calcd. for C ₂₄ H ₂ N ₂ 5F ₅ OSCs (M + Cs ^+): 602.0802. Found:

602,0818.s

Anal. calculated for C ₂₄ H ₂₅ F ₂ N ₅ OS. 0.5 H ₂ O. 0.1 EtOH: C, 60.16; H, 5.55; N, 14.49; S, 6.64. Found: C, 59.94; H, 5.24; N, 14.19;

S, 6.92.

* Example C (122) [4-Amino-2- (4-pyridin-4-yl-phenylamino) -thiazol-5-yl] - (2,6-difluoro-phenyl) -methanone

138

4-Pyridin-4-yl-aniline, which has the structural formula y3hCX ^nh2 was prepared first by the following procedure. A mixture of 4- (4-nitrophenyl) -pyridine (600 mg; 3.0 mmol; Wang et al., J.Phys.Chem., Vol 99 (1955), pages 6876-6888) and 10% Pd / C (100 mg) in EtOH (20 mL) was stirred under an atmosphere of hydrogen overnight. The catalyst was filtered off and the filtrate was concentrated in vacuo to give 510 mg (100% yield) of a white solid.

1 H NMR (CDCl ₃ ): δ 8.59 (2H, dd, J = 6.2, 1.6 Hz), 7.51 (2H, d, J =

8.6 Hz), 7.46 (2H, dd, J = 6.2, 1.6 Hz), 6.79 (2H, d, J = 8.6 Hz).

4- (4-Isothiocyanato-phenyl) -pyridine having the structural formula

was prepared as follows. To 4-pyridin-4-yl-aniline (200 mg, 1.18 mmol) in THE (35 mL) at 0 ° C was added excess Et ₃ N (0.33 mL, 2.4 mmol) and thiophosgene ( 99 μί, 1.29 mmol) dropwise. After 20 min at 0 ° C, then at ambient temperature for 10 min, the solvent was evaporated. The residue was suspended in water, filtered, washed with a little water, and dried under vacuum to give a brown solid, 240 mg (96% yield), which was used without further purification.

¹ H NMR (CDCl 3): δ 8.62 (2H, d, J = 6.3 Hz), 7.57 (2H, d, J = 8.6 Hz), 7.45 (2H, d, J = 6.3 Hz), 7.27 (2H, d, J = 8.6 Hz).

The title compound was prepared in a manner analogous to that used in Example C (1).

4- (4-Isothiocyanato-phenyl) -pyridine and 2-bromo-2 ', 6'-difluoro-acetophenone (from Example C (79)) gave, after recrystallization from EtOH, a brown powder in 64% yield, m.p. greater than 300 ° C.

¹ H NMR (DMSO-d ₆ ): δ 11.08 (1H, s), 8.61 (2H, d, J = 6.0 Hz), 8.25 (2H, s), 7.85 (2H , d, J = 8.8 Hz), 7.73 (2H, d, J = 8.8 Hz), 7.71 (2H, d, J = 6.0 Hz), 7.61-7 / 49 (1H, m), 7.23 (1H, d, J = 7.7Hz),

7.20 (1H, d, J = 8.1Hz).

• <!

Γ .139

HRFABMS ( ⁺ ): Calcd .: 409.0935. Found: 409.0921.

Anal. calculated for C ₂₁ H ₁₄ N ₄ OSF ₂ . 0.4 H ₂ O. 0.3 EtOH: C, 60.41; H, 3.90; N, 13.05; S, 7.47.

Found: C, 60.51; H, 3.65; N, 12.69; S, 7.86.

* Example C (123) / 4-Amino-2- / 4- (4-carbamoyl-piperazin-l-yl) phenylamino) -thiazol-5-yl / - (2,6-difluoro-phenyl) -methanone _{of NH2}

H 1-Carbamoyl-4- (4-nitrophenyl) piperazine having the structural formula

was prepared first by the method of Cain et al. _of J. Med. Chem., Vol. 20 (1977), pages 987-996, whereby 1- (4-nitrophenyl) piperazine was treated with potassium cyanate to give a white solid, 705 mg (99%), which was used without further purification.

NMR (CDCl 3): δ 6.71 (2H, d, J = 8.6 Hz), 6.50 (2H, d, J = 8.6 Hz), 5.97 (2H, bs), 4.58 (2H, bs), 3.39 (4H, dd, J = 5.1, 4.9 Hz), 2.82 (4H, dd, J = 5.1, 4.9 Hz).

1- (4-Amino-phenyl) -4-carbamoyl-piperazine, which has the structural formula was prepared as follows. A mixture of 4- (4-nitrophenyl) -piperazine-1-carboxylic acid amide (760 mg, 3.22 mmol), 10% Pd / C (120 mg), MeOH (20 mL), and THF (20 mL) was stirred under hydrogen for 2 hours.

The catalyst was filtered off and the filtrate was concentrated in vacuo

140 to give a white solid, 705 mg (99%), which was used without further purification.

Ή NMR (CDCl 3): δ 6.71 (2H, d, J = 8.6 Hz), 6.50 (2H, d, J = 8.6 Hz), 5.97 (2H, bs), 4.58 (2H, bs), 3.39 (4H, dd, J = 5.1, 4.9 Hz), 2.82 (4H, dd, J = 5.1, 4.9 Hz).

1-Carbamoyl-4- (4-structural formula of sothiocyanato-phenyl) -piperazine, which has been prepared by ZrC NCS as follows. Into suspension

1- (4-Amino-phenyl) -4-carbamoyl-piperazine (300 mg, 1.36 mmol) in THF (30 mL) at -35 ° C was gradually added dropwise triethylamine (0.38 mL, 2.73) mmol) and thiophosgene (104 μΐ, 1.36 mmol). The resulting light brown solid was filtered off, washed with a little water and dried under vacuum to give a brown powder. 337 mg (94% yield), which was used without further purification.

¹ H NMR (CDCl ₃ ): δ 7.08 (2H, d, J = 9.0Hz), 6.76 (2H, d, J = 9.0Hz), 4.45 (2H, bs), 3 50 (4H, dd, J = 5.4, 5.0 Hz), 3.15 (4H, dd, J = 5.4, 5.0 Hz).

The title compound was prepared in a manner analogous to that used in Example C (1). 1-Carbamoyl-4- (4-isothiocyanato-phenyl) -piperazine and 2-bromo-2 ', 6'-difluoro-acetophenone (from Example C (79)) gave a light gray powder in 45% yield, m.p. 278.5-279 ° C.

¹ H NMR (DMSO-d ₆ ): _δ 10.69 (1H, s), 8.16 (2H, bs), 7.63-7.51 (1H, m), 7.38 (2H, d, J = 9.0 Hz), 7.25 (1H, d, J = 7.8Hz), 7.21 (1H, d, J = 7.9Hz), 7.02 (2H, d, J = 9.0 Hz), 6.09 (2H, bs), 3.48 (2H, t, J = 4.7 Hz), 3.11 (2H, t, J = 4.7 Hz).

HRFABMS (M + Na): Calcd .: 81.1234. Found: 481.1246.

Anal. calculated for C ₂₁ H ₂₀ N ₆ O ₂ SF ₂ . 0.5 H ₂ O: C, 53.95; H, 4.53; N, r

141

17.98; S, 6.86. Found: C, 53.92; H, 4.35; N, 17.64; S, 6.64.

* Example C (124) [4-Amino-2- / 4- (3R, 4-dimethyl-piperazin-1-yl) -phenylamino) -thiazol-5-yl] - (2,6-difluoro-phenyl) -methanone

3R-Methyl-1- (4-nitrophenyl) piperazine having the structural formula

· Was prepared first as follows. (R) - (-) - 2-Methylpiperazine (186 mg, 1.86 mmol), 1-fluoro-4-nitrobenzene (131 mg, 0.93 mmol), Et ₃ N (0.26 mL, 1.86) mmol) and acetonitrile (2 mL) were refluxed overnight and concentrated in vacuo. The residue was suspended in water and the solid obtained was filtered, washed with a minimum amount of water and dried under vacuum to give a bright yellow solid, 128 mg (62% yield), which was used without further purification.

¹ H NMR (CDCl ₃ ): δ 8.12 (2H, d, J = 9.5 Hz), 6.82 (2H, d, J = 9.5 Hz), 3.80-3.71 (2H , m), 3.18-3.08 (1H, m), 3.04-2.88 (3H, m), 2.58 (1H, dd, J = 12.3 Hz), 1.16 ( 3H, d, J = 6.3Hz).

1,2R-Dimethyl-4- (4-nitro-phenyl) piperazine having the structural formula

was prepared as follows. A mixture of 3R-methyl-1- (4-nitro-phenyl) piperazine (124 mg, 0.56 mmol), sodium formate (93 mg, 1.37 mmol), formic acid (1.5 mL) and formalin (1, 5 mL) was stirred at ° C overnight, cooled, poured into ice / water and extracted

CHC1 ₃ . The organic layer was washed with brine, dried over Na ₂ SO ₄ and

Γ

142 concentrated to give 116 mg (71% yield) of yellow crystals which were used without further purification.

¹ H NMR (CD Cl ₃ ): δ 8.12 (2H, d, J) = 9.4 Hz), 6.82 (2H, d, J = 9.4) Hz), 3, 67 (IH, d, J = 12.4 Hz) 3.14 (IH, ddd, J = 12.4, 11.7, 1.5 Hz) r 2.90 (1 H, d, J = 11.7 Hz), 2.74 (IH, dd, J = 11.7, 10 Hz), 2, 40 (IH, m), 2.34 (3H, s); , 2.22 (IH, m), 1.16 (3 H, d, J = 6.3 Hz)

4- (3R, N-dimethyl-piperazin-l-yl) -aniline, which has the structural formula ^{H3C_N sl_} yD ^^ ^{'NH 2} C H ₃ was prepared as follows. A mixture of 1,2R-dimethyl-4- (4-nitrophenyl) -piperazine (168 mg, 0.71 mmol), 10% Pd / C (30 mg), and MeOH (10 mL) was stirred under hydrogen for 1.5 hours. clock. The catalyst was filtered off and the filtrate was concentrated in vacuo to give a dull yellow oil which was used without further purification.

¹ H NMR (CDCl ₃ ): δ 6.91 (2H, d, J = 8.8 Hz), 6.75 (2H, d, J = 8.8 Hz), 3.66-3.32 (4H) m), 3.05-2.89 (2H, m), 2.63-2.48 (2H, m),

2.44 (3H, s), 1.22 (3H, d, J = 6.1 Hz).

4- (4-Isothiocyanato-phenyl) -1,2R-dimethyl-piperazine having the structural formula

was prepared as follows. To excess 4- (3R, 4-dimethyl-piperazin-1-yl) -aniline (0.71 mmol) in THF (15 mL) at -35 ° C was added excess Et ₃ N (0.20 mL, 1 mL), 43 mmol) and thiophosgene (58 μΐ, 0.75 mmol) dropwise. The solvent was evaporated and the residue was partitioned with CHCl ₃ and water. The organic layer was dried with Na ₂ SO ₄ and concentrated to give a brown powder, 184 mg, which contained traces of Et ₃ N as determined by NMR, but was suitable for use without further purification.

142 / £ © © c © r r © i i r © c © © r r. <·· - · »

Γ f · Γ Γ - © ©

1 H NMR (CDCl ₃ ): δ 7.12 (2H, d, J = 9.1 Hz), 6.82 (2H, d, J = 9.1 Hz), 3.58-3.46 (2H, m), 3.13-3.03 (2H, m), 2.89-2.75 (1H, m), 2.65-2.41 (2H, m), 2.49 (3H, s) 1.27 (3H, d, J = 6.3 Hz).

The title compound was prepared in a manner analogous to that used in Example C (1).

4- (4-isothiocyanato-phenyl) -1,2R-dimethyl-piperazine a

2-bromo-2 ', 6'-difluoro-acetophenone (from Example C (79)) gave a yellow powder in 57% yield, m.p. 115-118 ° C.

¹ H NMR (DMSO-d ₆ ): 510.65 (1H, bs), 8.15 (2H, bs), 7.62-7.50 (1H, m), 7.35 (2H, d, J) = 9.0 Hz), 7.23 (1H, d, J = 7.7Hz), 7.20 (1H, d, J = 8.0Hz), 6.97 (2H, d, J = 9 0 Hz), 3.59-3.49 (2H, m), 3.34 (3H, s), 2.90-2.72 (2H, m), 2.40 (1H, t, J = 10.9 Hz), 2.28-2.05 (2H, m), 1.09 (3H, d, J = 6.2 Hz).

HRFABMS (MH ⁺ ): Calcd .: 444.1670. Found: 444.1656.

Anal. calculated for C ₂₂ H ₂₃ N ₅ OSF ₂ . 0.8 H ₂ O. 0.6 t-BuOH: C, 58.33; H, 6.14; N, 13.94; S, 6.38. Found: C, 58.38; H, 5.92; N, 13.89;

S, 6.33.

* Example C (125) [4-Amino-2- / 4- (3S, 4-dimethyl-piperazin-1-yl) -phenylamino) -thiazol-5-yl] - (2,6-difluoro-phenyl) -methanone

4- (4-isothiocyanato-phenyl) -1,2S-dimethyl-piperazine, which 'has the structure A ⁿ ~ ^ _ / - ⁷ · ^NCS - was prepared following the procedure employed for its enantiomer, 4- (4-isothiocyanato-phenyl) -1,2R-dimethyl-piperazine in Example C (124). Obtained yellow powder

143 yy yy yy yy yy yy y <y

144 showed a comparable NMR spectrum and was used without further purification.

The title compound was prepared in a manner analogous to that used in Example C (1).

4- (4-isothiocyanato-phenyl) -1,2S-dimethyl-piperazine and 2-bromo-2 ', 6'-difluoro-acetophenone (from Example C (79)) gave a yellow powder in 77% yield, m.p. Mp 110-116 ° C.

1 H NMR (DMSO-d 6): δ 10.65 (1H, bs), 8.15 (2H, bs), 7.62-7.50 (1H, m), 7.35 (2H, d, J = 9) 0 Hz), 7.23 (1H, d, J = 7.7Hz), 7.20 (1H, d, J = 8.0Hz), 6.97 (2H, d, J = 9.0 Hz), 3.59-3.49 (2H, m), 3.34 (3H, s), 2.90-2.72 (2H, m), 2.40 (1H, t, J = 10, 9 Hz), 2.28-2.05 (2H, m), 1.09 (3H, d, J = 6.2 Hz).

IR (KBr): 3386, 3274, 3168, 2970, 2807, 1620, 1589, 1547, 1517, 1464, 1429, 1238, 1001 cm &^lt; -1 & gt ^; .

HRFABMS (MH ⁺ ): Calcd .: 444.1670. Found: 444.1659.

Anal. Calcd. For C ₂₂ H ₂₃ N ₅ OSF ₂ . .0,7 ΗΌ. 0.2 t-BuOH: C, 58.15; H, 5.65; N, 14.87; S, 6.81. Found: C, 58.06; H, 5.61; N, 14.58;

S, 6.90.

* Example C (126) [4-Amino-2- / 4- [(3-dimethylamino-propyl) -methyl-amino] -phenylamino] -thiazol-5-yl] - (2,6-difluoro-phenyl) - methanone

h ₃ c

Well, he was prepared like

N- (4-Nitrophenyl) -N, N ', N'-trimethyl-propane-1,3-diamine having the structural formula

Γ

Γ

145 by a first procedure analogous to the tert-butyl / methyl (4-nitrophenyl) amino) acetate of Example C (103). 4-Fluoronitrobenzene and Ν, Ν, Ν'-trimethyl-propanediamine gave a yellow oil which was heated to 280 ° C at 1 torr to remove the starting materials to give an orange oil, 4.26 g (850 crude yield) which was used without further purification.

¹ H NMR (CDCl 3): 58.10 (2H, ddd, J = 9.5, 8.2, 5.3 Hz), 6.64 (2H, ddd, J = 9.5, 8.2, 5 3 Hz), 3.50 (2H, t, J = 7.2 Hz), 3.08 (3H, s), 2.07 (3H, t, J = 6.8 Hz), 2.23 ( 6H, s), 1.72-1.82 (2H, m).

N- (4-Aminophenyl) -N, N ', N'-propane-1,3-diamine having the structural formula

was prepared as follows. A mixture of N- (4-nitro-phenyl) -N, N ', N'-trimethyl-propane-1,3-diamine (1.72 g, 7.25 mmol) in stannous chloride (II) dihydrate (8.05) g, 36.2 mmol), dioxane (25 mL) and ethanol (5 mL) was heated under reflux for 3.5 h, then allowed to cool. Aqueous saturated Na ₂ CO ₃ was added to the obtained mixture until no gas evolution was observed. Celite was added to facilitate subsequent filtration. The solids were washed with MeOH and the filtrate was concentrated under reduced pressure and extracted with 10% MeOH / CHCl ₃ (4x). The combined extracts were washed with brine, dried over Na ₂ SO ₄ and evaporated to give a black oil which was purified by column chromatography with alumina (neutral, activity I) and 1% MeOH / CH ₂ Cl ₂ ) as eluent to give 0.39 g (26%) of a dark brown oil which was used without further purification.

¹ H NMR (CDCl ₃ ): δ 6.67 (4H, dd, J = 9.0, 8.6 Hz), 3.22 (2H, t, J = 7.2 Hz), 2.82 (3H) s), 2.31 (2H, t, J = 7.5Hz), 2.23 (6H, s), 1.70 (2H, p, J = 7.4Hz).

N- (4-isothiocyanato-phenyl) -N, N ', N'-trimethyl-propane-l, 3-diamine, which has the structural formula H ₃ crperrp, rrrr .. <- * · r, er ^r c ^r rrp ', ^r _r ^{f Γ Γ}

146 was prepared in a manner analogous to 4- (4-isothiocyanato-phenyl) -1,2R-dimethyl-piperazine from Example C (124).

N- (4-Aminophenyl) -N, N ', N'-propane-1,3-diamine gave a black oil in 86% yield of the raw material which was used without further purification.

¹ H NMR (CDCl 3): δ 7.09 (2H, d, J = 9.0 Hz), 6.59 (2H, d, J = 9,

Hz), 3.38 (2H, J = 7.2 Hz), 2.94 (3H, s), 2.36 (2H, t, J = 7.2 Hz), 2.29 (6H, s) 1.78 (2H, p, J = 7.2 Hz).

IR (KBr): 2127, 1605, 1514, 1379 ^cm-first

The title compound was prepared in a manner analogous to that used in Example C (1).

N- (4-Isothiocyanato-phenyl) -N, N ', N'-trimethylpropane-1,3-diamine and 2-bromo-2', 6'-difluoro-acetophenone (from Example C (79)) gave a brown oil which was purified by flash column chromatography in a step gradient of 7-14% MeOH / CHCl 3 and precipitated from CH ₂ Cl ₂ / hex to give an amorphous yellow solid in 51% yield, mp 115-120 ° C (compound).

¹ H NMR (DMSO-d ₆ ): δ 10.50 (1H, bs), 8.05 (2H, bs), 7.50 (1H, ddd, J = 15.3, 8.4, 6.7 Hz), 7.10-7.35 (4H, m), 6.68 (2H, d, J = 9.1)

Hz), 2.84 (3H, S), 2.27 (2H, t, J = 7.2 Hz), 2.16 (6H, s), 1.61 (2H, p, J = 7.3 Hz) ).

IR (KBr): 3393, 3279, 3165, 2951, 1619, 1545, 1524, 1462, 1436 cm &^lt; -1 ^&gt;.

HRFABMS: Calcd. for C ₂₂ H _2i F ₂ N ₅ OS (MH +): 446.1826. Found:

446.1810.

Anal. calculated for C ₂₁ H ₂₃ F, N ₅ CS. 0.8 H ₂ 0. 0.4 C ₆ H ₁₄ : C, 59.28; H, 6.56; N, 14.16; S, 6.49. Found: C, 59.37; H, 6.31; N, 13.76;

S, 6.26.

♦ Example C (127) (2,6-Difluoro-phenyl) - [2- [4- (4-pyridin-4-yl-piperazin-1-yl) -phenylamino] -thiazol-5-yl] -methanone

147

0-0

1- (4-Nitro-phenyl) -4-pyridin-4-yl-piperazine, which has the structural formula - was prepared according to the procedure

NO, analogous to the target. butyl (methyl- (4-nitro-phenyl) -amino) acetate of Example C (103). 4-Fluoronitrobenzene and 1- (4-pyridyl) piperazine (Ratous et al., J. Med. Chem., Vol. 8 (1965), pp. 104-107) gave a brown powder in 27% yield which was used without further purification.

¹ H NMR (CD ₃ OD): δ 8.20 (2H, d, J = 5.0Hz), 8.08 (2H, d, J = 9.4Hz), 7.04 (2H, d, J) = 9.5 Hz), 3.62-3.68 (4H, m), 3.50-3.56 (4H, m).

4- (4-Pyridin-4-yl-piperazin-1-yl) -aniline, structural formula a.

-nh ₂ which has been prepared in a manner analogous to 4- (4-methyl-piperazin-1-yl) -aniline of Example C (70). 1- (4-Nitro-phenyl) -4-pyridin-4-yl-piperazine gave a brown powder in 100% crude yield, which was used without further purification.

^lT H NMR (CD3OD): δ 8.12 (2H, d, J = 6.7 Hz) ,. 6.90 (4H, bd, J = 8.9 Hz), 6.74 (2H, d, J = 6.6 Hz), 3.56 (4H, dd, J = 5.3, 5.0) Hz), 3.14 (4H, dd, J = 5.0, 4.2 Hz).

1- (4-Isothiocyanato-phenyl) -4-pyridin-4-yl-piperazine, which has the structural formula (I), was prepared as follows. To a solution of 4- (4-pyridin-4-yl-piperazin-1-yl) aniline (2.00 g, 7.86 mmol) in 10% aqueous HCl (1 mL) was added

148 added thiophosgene (720 μΐ, 9.43 mmol). After 0.5 h, the resulting yellow precipitate was filtered, washed with saturated aqueous NaHCO ₃ and H ₂ O and dried under high vacuum to give 1.9 g (82% yield) of a yellow powder which was used without further purification.

¹ H NMR (DMSO-d ₆ ): δ 6.73 (4H, d, J = 8.8 Hz), 6.51 (4H, d, J = 8.8 Hz), 3.32 (4H, bs 3.29 (4H, bs).

The title compound was prepared as described in Example C (1). 1- (4-Isothiocyanato-phenyl) -4-pyridin-4-yl-piperazine and 2-bromo-2 ', 6'-difluoro-acetophenone (from Example C (79)) gave after recrystallization with traces of DMSO in MeOH / CHCl ₃ light brown powder in 30% yield, mp 155-157 ° C.

¹ H NMR (DMSO-d 6): δ 8.16 (2H, d, J = 6.0 Hz), 8.04 (1H, bs), 7.40-7.52 (1H, m), 7, 32 (2H, d, J = 8.7Hz), 7.15 (2H, t, J = 7.7Hz), 6.96 (2H, d, J = 9.0Hz), 6.85 ( 2H, d, J = 5.5 Hz), 3.60 (4H, bs).

HRFABMS: Calcd. for C ₂₅ H ₂₃ F ₂ N ₆ OS (MH ⁺ ): 493.1622. Found:

493.1606.

Analyt. for C ₂₅ H ₂₃ F ₂ N ₆ OS. 0.7 MeOH. 0.1 CHCl ₃ . 0.1 DMSO: C, 58.40; H, 4.81; N, 15.72; S, 6.60. Found: C, 58.38; H,

4.50; N, 15.37; S, 7.00.

* Example C (128) [4-Amino-2- / 4- (1-methyl- [1,4] -diazepan-4-yl) -phenylamino] -thiazol-5-yl] - (2,6-difluoro) (phenyl) -methanone

1-Methyl-N- (4-nitrophenyl) - [1,4] diazepane, which has the structural formula

R r rc r γ

R Τ 'r r c c γ β Γ Γ Γ ř “· · - r Γ» γ, -1 * e

149 was prepared in a manner analogous to the tert-butyl / methyl (4-nitro-phenyl) -amino / acetate of Example C (103). 1-Methyl-homopiperazine gave a yellow powder in 93% yield, which was used without further purification.

¹ H NMR (CDCl ₃ ): δ 8.12 (2H, d, J = 9.5Hz), 6.64 (2H, d, J = 9.5Hz), 3.56-3.70 (4H, m), 2.74 (2H, dd, J = 4.9, 3.3 Hz), 2.58 (2H, dd, J = 4.9, 3.3 Hz), 2.58 (2H, dd) J = 5.6, 5.4 Hz), 2.40 (3H, s), 2.00-2.08 (2H, m).

4- (4-Methyl- [1,4] diazepan-1-yl) aniline having the structural formula

was prepared in a manner analogous to 4- (4-methyl-piperazin-1-yl) aniline of Example C (70). 1-Methyl-4- (4-nitro-phenyl) - [1,4] diazepane gave a violet oil in 100% crude yield, which was used immediately without further purification.

¹ H NMR (CDCl 3): δ 6.68 (2H, d, J = 12.2 Hz), 6.60 (2H, d, J = 6.8 Hz), 3.52 (2H, dd, J = 4.8, 4.7 Hz), 3.43 (2H, t, J = 6.3 Hz), 2.71 (2H, dd, J = 4.9, 4.7 Hz), 2.58 ( 2H, dd, J = 5.5, 5.4 Hz), 2.38 (3H, s), 1.95-2.04 (1H, m).

1- (4-Isothiocyanato-phenyl) -4-methyl- [1,4] diazepane, which has the structural formula

was prepared in a manner analogous to 1- (4-isothiocyanato-phenyl) -1H-imidazole of Example C (41). 4- (4-Methyl- / l, 4 / diazepan-yl) -aniline gave a crude product, which was extracted with CHC1 ₃ to yield a black oil in 85% yield raw material, which was used immediately without further purification.

¹ H NMR (CDCl 3): δ 7.02 (2H, d, J = 9.0 Hz), 6.56 (2H, d, J = 9.0 Hz), 3.54 (2H, dd, J = 4.8, 4.8 Hz), 3.45 (2H, t, J = 6.3 Hz),

150

2.67 (2H, dd, J = 4.9, 4.8 Hz), 2.53 (2H, dd, J = 5.6, 5.4 Hz), 2.36 (3H, s), 1 97 (2H, p, J = 5.7Hz).

The title compound was prepared in a manner analogous to that used in Example C (1).

1- (4-Isothiocyanato-phenyl) -4-methyl- [1,4] diazepine a

2-bromo-2 ', 6'-difluoro-acetophenone (from Example C (79)) gave, after crystallization from boiling EtOH, a light brown powder in 26% yield, m.p. 138-140 ° C.

Ή NMR (DMSO-d ₆ ): δ8.05 (1H, bs), 7.42-7.52 (1H, m), 7.10-7.22 (4H, m), 6.64 (2H, d, J = 9.1 Hz), 3.36-3.52 (4H, m), 2.58 (2H, dd, J = 4.8, 4.7 Hz), 2.42 (2H, dd) J = 6.6 (5.4 Hz), 2.25 (3H, s), 1.82-1.92 (2H, m).

HRFABMS: Calcd. for C ₂₂ H ₂₄ F ₂ N ₅ OS (MH +): 444.1670. Found: 444.1656.

Anal. calculated for C ₂₂ H ₂₃ F ₂ N ₅ OS. 0.5 H ₂ O. 0.8 EtOH: C, 57.92; H, 5.93; N, 14.31; S, 6.55. Found: C, 58.05; H, 5.69; N, 14.15;

S, 6.55.

♦ Example C (129)

3- (4- (4-Amino-5- (2,6-difluorobenzoyl) -thiazol-2-yl-amino) -phenyl) -methylamino) -propionitrile

3- / Methyl (4-nitro-phenyl) amino / -propionitrile, which has the structural formula _IIa C was prepared following NC ^ Yes ₂ procedure. Benzyltrimethylammonium hydroxide (7.23 ml of a 40% solution in

MeOH) was added to a suspension of N-methyl-4-nitroaniline (5.00 g,

32.9 mmol) and acrylonitrile (7.23 mL) in dioxane (80 mL). The resulting solution was heated at 55 ° C for 3.5 h, then poured onto water

Γ

F · f

Γ r

And extracted with 20% isopropanol in chloroform. The separated organic layer was washed with water, dried over K ₂ CO ₃ and concentrated to a slurry of yellow solid which was diluted with ether. The solid was filtered and dried under vacuum to give 6.15 g (91% yield) of a yellow solid, which was used without further purification.

¹ H NMR (CDCl ₃ ): δ8.17 (2H, d, J = 9.4 Hz), 6.66 (2H, d, J = 9.4 Hz), 3.82 (2H, t, J = 6.7 Hz), 3.19 (3H, s), 2.66 (2H, t, J = 6.7 Hz).

3 - [(4-Amino-phenyl) -methyl-amino] propionitrile having the structural formula was prepared

NcT '' ΌχΗ ₂ t

by a procedure analogous to

4- (3S, 4-Dimethyl-piperazin-1-yl) -phenylamine of Example C (134). 3- (Methyl- (4-nitro-phenyl) -amino) -propionitrile gave a brown oil in 100% yield, which was used without further purification.

¹ H NMR (CDCl ₃ ): δ 6.68 (4H, s), 3.57 (2H, t, J = 7.0Hz), 2.90 (3H, s), 2.51 (2H, t , J = 7.0 Hz).

3- [(4-Isothiocyanato-phenyl) -methyl-amino] -propionitrile having the structural formula - was prepared

in a way analogous to ·

4- (4-isothiocyanato-phenyl) -1,2S-dimethyl-piperazine in Example C (134). 3 - [(4-Amino-phenyl) -methyl-amino] -propionitrile gave a brown solid in 95% yield, which was used without further purification.

¹ H NMR (CDCl ₃ ): δ 7.15 (2H, d, J = 9.1 Hz), 6.62 (2H, d, J = 9.1 Hz), 3.72 (2H, t, J = 6.8 Hz), 3.05 (3H, s), 2.58 (2H, t, J = 6.8 Hz).

The title compound was prepared according to the procedure

0

152 'analogous to the procedure used in Example C (1).

3- [4- (4-Isothiocyanato-phenyl) -methylamino] -propionitrile a

2-bromo-2 ', 6'-difluoroacetophenone (from Example C (79)) afforded a morph yellow powder in 66% yield, m.p. 120-130 ° C (dec.).

¹ H NMR (DMSO-d ₆ ): δ 10.52 (1H, bs), 8.10 (2H, bs), 7.49 (1H, ddd, J = 15.3, 8.2, 6.7 Hz), 7.26 (2H, d, J = 8.2 Hz), 7.15 (2H, dd, J = 8.1, 7.7 Hz), 6.76 (2H, d, J = 9 Hz) 1 Hz), 3.63 (2H, t, J = 6.7 Hz), 2.91 (3H, s), 2.69 (2H, t, J = 6.7 Hz).

IR (KBr): 3417, 3.9, 1618, 1548, 1523, 1463, 1436, 1376, 1356, 1234, 1001 cm ^-1 .

HRFABMS: Calcd. for C ₂₀ H ₁₇ N ₅ OSF ₂ Na (M + Na ⁺ ): 436.1020.

Found: 436.1030.

Anal. calculated for C ₂₀ H ₁₇ N ₅ OSF ₂ . 0.2 H ₂ O. 0.45 t-BuOH: C, 5.13; H, 4.90; N, 15.55; S, 7.12.

Found: C, 57.88; H, 4.79; N, 15.16; S, 6.95.

* Example C (130)

2- (4-Amino-2- (4-nitro-phenylamino) -thiazole-5-carbonyl) -phenyl benzoate o ₂ n.

nh ₂

AND

The title compound was prepared essentially as described in Example C (1). Further, two reaction products were isolated after flash column chromatography and the characteristics for (Z) - and (E) -4- (2-hydroxyphenyl) -3H-thiazol-2-ylidene-cyanamide were identified as follows. 4-Nitrophenylisothiocyanate and 2'-benzoyloxy-2-bromoacetophenone gave the title compound as a yellow solid, mp: 258-260 ° C.

¹ H NMR (DMSO-d ₆ ): δ 11.35 (1H, s), 8.23 (2H, d, J = 9.3 Hz),

7.88-8.04 (4H, m), 7.85 (2H, d, J = 9.2 Hz), 7.36-7.67 (1H, m),

153 y * y y y y y, y, ·· κ y y y

7.52-7.63 (4H, m); 7.93-7.45 (2H, m).

¹³ C NMR (MeOH-dJ: δ 181.5, 166.4, 164.4, 147.2, 145.8, 142.0, 135.2, 134.3, 131.2, 130.0, 129 , 3, 129.2, 128.3, 126.5, 125.6, 123.9, 118.3.

Anal. calc. for C ₂₅ H _U N ₄ O ₅ S: C, 59.99; H, 3.50; N, 12.17; S, 6.96. Found: C, 58.25; H, 3.54; N, 11.77; S, 6.94.

The compound that eluted earlier, (Z) -4- (2-hydroxy-phenyl) -3- (4-nitrophenyl) -3H-thiazol-2-ylidene-cyanamide, having the structural formula was

isolated as a yellow amorphous substance.

Ή NMR (DMSO-d ₆ ): δ 9.79 (1H, s), 8.18 (2H, d, J = 9.0 Hz), 7.55 (2H, d, J = 9.0 Hz) 7.26 (1H, dd, J = 7.5, 1.5 Hz), 7.17 (1H, ddd, J = 7.5, 7.4, 1.5 Hz), 7.05 (1H , s), 6.79 (1H, dd, J = 7.6, 7.4 Hz), 6.65 (1H, d, 8.2 Hz). ¹³ C NMR (MeOH-d ₄ ): δ 176.8, 157.9, 150.3, 143.6, 141.7, 134.3, 134.2, 132.0, 126.0, 122.1, 119.5, 119.3, 117, 9, 107.3.

HRFABMS: Calcd. for C ₁₅ H ₁₀ N ₄ O ₃ S (MH ⁺ ): 339.0552. Found: 339.0550.

A compound that eluted later, (E) -4- (2-hydroxy-phenyl) -3- (4-nitrophenyl) -3H-thiazol-2-ylidene-cyanamide, having the structural formula

was isolated as a yellow amorphous substance.

¹ H NMR (DMSO-d 6): δ 13.2 (1H, s), 8.25 (2H, d, J = 9.2 Hz), 7.55 (1H, dd, J = 7.8, 1) 5 Hz), 7.55 (1H, ddd, J = 8.6, 7.5, 1.1 Hz), p ρ pp

154 ··

7.41 (1H, ddd, J = 8.6, 7.5, 1.1 Hz), 7.25 (1H, dd, J = 8.1, 1.0 Hz), 7.13 (1H, d, 9.2 Hz), 7.01 (1H, s).

1 C NMR (MeOH-dJ: δ 174.8, 162.1, 152.2, 143.6, 134.0, 131.4, 129.9, 126.4, 126.2, 122.0, 121 , 5, 117.8, 105.6.

ESIMS: Calcd. For C ₁₆ H ₁₀ N ₄ O ₃ S (MH ⁺ ): 339. Found: 339.

* Example C (131):

(4-Amino-2- / 4- / 4- (2,2,2-trifluoroethyl) -piperazin-1-yl) -phenylamino-thiazol-5-yl) -2,6-difluoro-f enyl) -methanone

1- (4-Nitrophenyl) -4- (2,2,2-trifluoroethyl) -piperazine having the structural formula no ₂ was prepared in a manner analogous to tert-butyl / methyl- (4-nitrophenyl) amino-acetate from of Example C (103). 1- (4-Nitro-phenyl) -piperazine a

1,1,1-trifluoro-2-iodoethane gave a yellow-orange solid in 33% crude yield.

1 H NMR (CDCl 3): δ 8.13 (2H, d, J = 9.2 Hz), 6.82 (2H, d, J = 9.2 Hz), 3.51-3.38 (4H, m 3.10-2.99 (2H, m), 2.87-2.77 (4H, m).

4- (4- (2,2,2-Trifluoro-ethyl) -piperazin-1-yl) -aniline having the structural formula NH, the prepared -aniline from the example procedure was then analogous to 4- (4-methyl- piperazin-1-yl) C (70).

Γ IΓ # *

1- (4-Nitro-phenyl) -4- (2,2,2-trifluoro-ethyl) -piperazine gave a pale brown solid in 100% crude yield.

155 ·

Hz).

1- (4-Isothiocyanato-phenyl) -4- (2,2,2-trifluoro-ethyl) piperazine having the structural formula was prepared in a manner analogous to

1- (4-isothiocyanato-phenyl) -4-methyl-piperazine from Example C (70) from 4- [4- (2,2,2-trifluoro-ethyl) -piperazin-1-yl] -aniline, to give of brown powder in 89% yield.

¹ H NMR (CDCl 3): δ 7.15 (2H, d, J = 9.1 Hz), 6.85 (2H, d, J = 9.0 Hz), 3.25 (4H, dd, J = 4.9, 5.2 Hz), 3.05 (2H, g, J = 9.5 Hz),

2.86 (4H, dd, J = 5.1, 4.8 Hz).

The title compound was prepared as described in Example C (1).

1- (4-Isothiocyanato-phenyl) -4- (2,2,2-trifluoro-ethyl) -piperazine; and

2-bromo-2 ', 6'-difluoroacetophenone (from Example C (79)) gave, after purification by column chromatography with 5% MeOH / CHCl ₃ as eluent, a yellow powder in 63% yield, mp 99-102 ° C.

1 H NMR (DMSO-d ₆ ): δ 8.12 (1H, bs), 7.58-7.46 (1H, m), 7.30 (2H, bd J = 7.4 Hz), 7.18 (2H, dd, J = 7.8, 7.7 Hz), 6.92 (2H, d, J = 8.9 Hz), 3.24 (2H, q, J = 10.3 Hz), 3 12 (4H, dd, J = 4.1, 5.0 Hz), 2.76 (4H, bd, J = 4.6 Hz).

IR (KBr): 3394, 3276, 3178, 3058, 2954, 2829, 1617, 1588, 1547, 1462, 1426, 1231 cm ^-1 .

Anal. calc. for C ₂₂ H ₂ OF ₂ N ₅ OS. 0.15 CHC1 _3: C, 51.62; H, 3.94; N, 13.59; S, 6.22. Found: C, 51.68; H, 3.93; N, 13.39; S, 6.03.

r r

156 * Example D (1):

(3-Aminophenyl) - (4-amino-2-phenylamino-thiazol-5-yl) methanone

A mixture of the title compound of Example A (1) ((4-amino-2-phenylamino-thiazol-5-yl) - (3-nitrophenyl) -methanone, 520 mg, 1.53 mmol) and 10% palladium on carbon (80 mg) in THF (10 mL) was stirred under an atmosphere of hydrogen overnight. The catalyst was filtered off and the filtrate was concentrated in vacuo to give 470 mg of a crude solid which was recrystallized from ethyl acetate / benzene to give 100 mg (19% yield) of a pale yellow powder, m.p. Mp 162-164 ° C.

¹ H NMR (DMSO-d ₅ ): δ 10.75 (1H, s), 8.42 (2H, bs), 8.15 (2H, bs), 7.60 (2H, d, J = 7, 8 Hz), 7.34 (2H, d, J = 7.8 Hz), 7.23 (1H, t,

J = 7.8 Hz), 7.14 (1H, s), 7.07 (IH, d, J = 7.8 Hz), 7.05 (IH, t, J = 7.8 Hz), 6 91 (1H, d, J = 7.8Hz).

FABMS (MH ^&lt; ^{+ &gt;} ): 311.

Anal. calculated for C ₁₆ H ₁₄ N ₄ OS. H ₂ O. C ₆ H ₆ : C, 59.30; H, 4.98; S, 9.54. Found: C, 59.02; H, 4.61; N, 16.34; S, 9.25.

* Example D (2):

(4-Amino-phenyl) - (4-amino-2-feriylamino-thiazol-5-yl) -methanone

o nh ₂

The title compound was prepared as described in Example D (1). Catalytic reduction of the compound of

157 title of Example Α (2) ((4-nitro-phenyl) - (4-amino-2-phenylaminothiazol-5-yl) -methanone) gave, after recrystallization from ethanol, 410 mg (90% yield) of a red amorphous powder, tt greater than 300 ° C.

¹ H NMR (DMSO-d ₆ ): δ 10.85 (1H, bs), 8.44-8.20 (2H, bs), 8.36 (1H, d, J = 8.7 Hz), 8 17 (1H, d, J = 8.7Hz), 7.89 (1H, d, J = 15.9Hz), 7.62 (2H, d, J = 7.8Hz), 7.37 (2H, t, J = 7.8Hz), 7.09 (1H, t, J = 7.8Hz).

FABMS (MH ^&lt; ^{+ &gt;} ): 311.

Anal. calcd for C _1S H ₁₄ N ₄ OS. 0.5 H ₂ O: C, 60.17; H, 4.73; N,. 17.54; S, 10.04. Found: C, 60.09; H, 4.73; N, 17.58; S, 9.93. * Example D (3):

/ 4-Amino-2- (4-dimethylamino-ethylamino-phenylamino) -thiazol-5-yl / - (2-aminophenyl) -methanone H ₃ C <Pn ₃ H _and Z \

nh ₂

The title compound was prepared as described in Example D (1). Catalytic reduction of the title compound of Example C (4) gave 26 mg (30% yield) of an amorphous solid.

¹ H NMR (DMSO-d 6): δ 10.38 (1H, s), 8.06 (2H, bs), 7.31 (2H, d, J = 9.0 Hz), 7.30 (1H, s, d, J = 7.5 Hz), 7.08 (1H, t, J = 7.5 Hz), 6.72 (2H, d, J = 9.0 Hz), 6.68 (1H, d, J = 7.5 Hz), 6.51 (1H, t, J = 7.5 Hz), 5.75 (2H, s), 2.88 (6H, s).

FABMS (MH ^&lt; ^{+ &gt;} ): 354.

Anal. calculated for C ₁₈ H ₁₉ N ₅ OS. 0.5 H ₂ 0. 0.3 MeOH: C, 59.07; H, 5.74; N, 18.82; S, 8.62. Found: C, 59.2.4; H, 5.56; N, 18.51; S, 8.36.

re rrrrrrrrr river rrcrr ^rr

158 * Example D (4):

(4-Amino-2- (4-amino-phenylamino-thiazol-5-yl) -phenyl-methanone)

The title compound was prepared as described in Example D (1). Catalytic reduction of the title compound of Example A (8) (i.e., 4-amino-2- (4-nitrophenylamino) thiazol-5-yl) -phenylmethanone, 450 mg, 1.32 mmol) gave, after recrystallization from ethanol 120 mg (29% yield) of an orange powder, m.p. Mp 167-169 ° C.

¹ H NMR (DMSO-d ₆ ): δ 10.38 (1H, bs), 8.15 (2H, bs), 7.64-7.55 (2H, m), 7.47-7.38 ( 3H, m), 7.10 (2H, d, J = 8.6 Hz), 6.55 (2H, d, J =

8.6 Hz), 5.20 (2H, bs).

FABMS (MHU: 311H);

Anal. calculated for C ₁₆ H ₁₄ N ₄ OS. H ₂ O: C, 56.96; H, 5.08; N, 16.61;

S, 9.50. Found: C, 56.94; H, 5.07; N, 16.60; S, 9.64.

* Example D (5):

4- (4-Amino-5- (3-amino-5-amino-thiophene-2-carbonyl) -thiazol-2-ylamino) -benzenesulfonamide

nh ₂

The title compound was prepared in a manner analogous to that used in Example D (1). The title compound of Example C (95) was hydrogenated and recrystallized from EtOH to give a brown powder in 96% yield, m.p. M.p. 268-271 ° C.

Ή NMR (DMSO-d 6): δ 10.97 (1H, s), 7.91 (2H, s), 7.82 (2H, d, J = <

Γ ©

© Γ yyyy © © yy yy yy

Γ © Γ © © <'© Γ rr © r'·' ^r © '· - © rrrr · r ^r

159 '· ·

9.1 Hz), 7.78 (2H, d, J = 9.1 Hz), 7.28 (2H, s), 6.43 (2H, s),

5.81 (1H, s); 2.34 (3H, s).

FABMS (MH ^&lt; ^{+ &gt;} ): 410.

Anal. calculated for C ₁₅ H ₁₅ N ₅ O ₃ S ₃ . 0.1 H ₂ O. 0.3 EtOH: C, 44.07; 4,0, 4.03; N, 16.47; S, 22.63. Found: C, 44.23; H, 3.93; N, 16.07; S, 23.01.

* Example E (l):

4- (4-Amino-5- (2-nitro-benzoyl) -thiazol-2-ylamino) -benzoic acid

H

To a suspension of the title compound of Example A (5) (i.e., ethyl 4- / 4-amino-5- (2-nitro-benzoyl) -thiazol-2-ylamino) -benzoate, 950 mg, 2.3 mmol) in methanol - (15 mL), 3N NaOH (10 mL) was added. After 30 minutes, the mixture was acidified to pH 4 with 1N HCl, forming a yellow precipitate. The mixture was diluted with water (100 mL). The solid was filtered off and washed with water. Recrystallization from ethanol gave 672 mg (76% yield) of yellow crystals, m.p. 286-292 [deg.] C.

1 H NMR (DMSO-d ₆ ): δ 12.75 (1H, s), 11.13 (1H, s), 8.12 (2H, bs), 8.08 (12H, d, J = .7, 8 Hz), 7.91 (2H, d, J = 8.7 Hz), 7.82 (1H, td, J = 8.4, 0.9 Hz), 7.78-7.68 (4H, m).

FABMS (MH ^&lt; ^{+ &gt;} ): 385.

Anal. calcd for C ₁₉ H ₁₈ N ₄ O ₃ S: C, 53.12; H, 3.15; N, 14.58; S, 8.34. Found: C, 53.29; H, 3.25; N, 14.31; S, 8.11.

* Example E (2):

4- / 4-Amino-2- (4-sulfamoyl-phenylamino) -thiazole-5-carbonyl / benzoic acid

0 r r 0

160 acid

To a suspension of ethyl 4- [4-amino-2- (4-sulfamoyl-phenylamino) -thiazole-5-carbonyl] -benzoate (500 mg, 1.12 mmol; Example C (34)) in MeOH (10 mL) was added 1 N aqueous NaOH (3.4 mL, 3.4 mmol) was added. After 4 hours, the resulting mixture was acidified with 1N aqueous HCl to pH 3 and filtered. The isolated brown solid was crystallized in EtOH to give 330 mg (70% yield) of light brown crystals, m.p. 298.5-300 ° C.

1 H NMR (DMSO-d ₅ ): δ 13.15 (1H, s), 11.14 (1H, s), 8.31 (2H, bs), 8.02 (2H, d, J = 8, 1 Hz), 7.78 (4H, s), 7.77 (2H, d, J = 8.1 Hz), 7.26 (2H, s).

FABMS (M + Na ⁺ ): Calcd .: 441.0303. Found: 441.0320.

Anal. calculated for C ₁₇ H ₁₄ N ₄ O ₅ S ₂ . 0.4 H ₂ O: C, 47.97; H, 3.50; N, 13.16; S, 15.07. Found: C, 48.04; H, 3.48; N, 12.98; S, 15.18.

* Example F:

2- / 4-Amino-2- (4-methoxy-phenylamino) -thiazole-5-carbonyl / ₃ c h -benzonitrile

To a solution of the compound of Example 0 (12) (2.00 g, 4.43 mmol) in pyridine (5 mL) was added copper (I) cyanide (709 mg, 8.86 mmol), and the mixture was heated to reflux. After 2 hours, the resulting mixture was allowed to cool, acidified with 1N aqueous HCl, and rt rt

Extracted with 20% MeOH / GHCl ₃ . The CHCl ₃ extracts were combined, washed with H ₂ O and brine, dried over Na ₂ SO ₄ and evaporated to give a dark brown viscous oil which was purified by preparative thin layer chromatography with 5% MeOH / CH ₂ Cl ₂ and precipitated from EtOH to afford 255 mg (61% yield) of a yellow amorphous solid which decomposed at 110-116 ° C.

¹ H NMR (DMSO-d ₆ ): δ 10.70 (1H, s), 8.24 (2H, bs), 7.91 (1H, d, J = 7.8 Hz), 7.80-7 66 (2H, m), 7.61 (1H, td, J = 7.8, 1.2 Hz), 7.42 (2H, d, J = 9.0 Hz), 6.92 (2H, d, J = 9.0 Hz), 3.72 (3H, s).

FABMS (MH ^&lt; ^{+ &gt;} ): 351.

Anal. calculated for C ₁₈ H ₁₄ N ₄ O ₂ S. 0.25 H ₂ O. 0.2 EtOH: C, 60.69; H, 4.35; N, 15.39; S, 8.81. Found: C, 60.84; H, 4.24; N, 15.07;

S, 9.02.

* Example G:

(4-Amino-2- (1H-benzoimidazol-6-ylamino) -thiazol-5-yl) - (3-amino-2,3-dichlorophenyl) methanone

The title compound of Example C (82),

N- (3- / 4-amino-2- (1H-benzoimidazol-6-ylamino) -thiazole-5-carbonyl / 2,4-dichlorophenyl) acetamide (100 mg, 0.220 mmol) was taken up in 6N aqueous HCl ( 4 ml) and stirred at ambient temperature for 24 hours.

The mixture was adjusted to pH 7 with 2N aq. NaOH and the resulting pale yellow precipitate was filtered off, washed with water, recrystallized from MeOH / H ₂ O and dried under high vacuum. The yellow solid was obtained in 36% yield, mp 235-237 ° C.

^; 1 H NMR (DMSO-d 6): δ 8.16 (1H, bs), 7.86 (2H, bs), 7.38-7.62 (1H, m), 7.18 (1H, d, J = 8.5 Hz), 7.02 (1H, d, J = 8.8 Hz), 6.68 (1H, rercrrrerr)

162 (d, J = 8.7 Hz), 5.50 (1H, bs).

IR (KBr): 3177, 1614, 1543, 1443, 1308 cm &^lt; -1 & gt ^; .

FABMS (MH ^&lt; ^{+ &gt;} ): 419.

Anal. calculated for C ₁₇ H ₁₂ Cl ₂ N ₆ OS. 0.8 H ₂ O. 1 MeOH: C, 46.42; H, 3.81; N, 18.04; S, 6.88. Found: C, 46.37; H, 3.45; Cl, 15.29; N, 17.84; S, 6.77.

* Example H (l):

(4-Amino-2- (4-piperazin-1-yl-phenylamino) -thiazol-5-yl) - (3-methyl-thiophen-2-yl) -methanone trihydrochloride

H

The title compound was prepared as follows. To a solution of the title compound of Example C (104) (100 mg, 0.20 mmol) in a mixture of THF (1 mL) and MeOH (0.5 mL) was added a solution of 4N HCl in dioxane (200 μΐ, 0.80). mmol). The resulting suspension was heated under reflux for 2 hours. The suspension was allowed to cool and then filtered. The isolated solid was washed with anhydrous ether and dried to give a yellow solid in 97% yield, m.p. Mp 198-200 ° C.

¹ H NMR (DMSO-d ₆ ): δ 10.80 (1H, m), 9.22 (1H, bs), 7.60 (1H, d, J 5.0 Hz), 7.42 (1H, d, J = 8.7 Hz), 6.98-7.08 (3H, m), 3.38 (4H, d, J = 4.4 Hz), 3.22 (4H, s), 2, 18 (3 H, s).

IR (KBr): 3177, 1614, 1543, 1443, 1308 ^cm-first

HRFABMS: Calculated for C ₁₉ H ₂₂ N ₅ OS ₂ (MH ^+): 400.1266. Found: 400.1254.

Anal. calculated for Ο ₁₉ Η ₂₁ Ν ₅ Ο3 ₂ . 0.6 H ₂ 0. 3 HCl: C, 43.91; H, 4.89 N, 13.47; S, 12.34. Found: C, 43.61; H, 4.97; N, 13.12; S, rr

12.16.

* Example H (2):

(J-Amino-2,6-dichloro-phenyl) - [4-amino-2- (4-piperazin-1-yl-phenylamino) -thiazol-5-yl] -methanone trihydrochloride.

hh ₂ n

The title compound was prepared as described in Example H (1). The title compound of Example C (106) gave a yellow solid in 48% yield, m.p. greater than 280 ° C.

¹ H NMR (DMSO-d ₆ ): δ 8.88 (1H, bs), 8.00 (1H, bs), 7.40 (2H, bs), 7.18 (1H, d, J = 8, 7 Hz), 6.98 (2H, d, J = 8.4 Hz). 6.80 (1H, d, J = 8.7 Hz), 3.38 (4H, s), 3.12 (4H, s).

IR (KBr): 3406, 1618, 1560, 1458, 1308 cm &^lt; -1 ^&gt;.

HRFABMS: Calcd for C ₂₀ H ₂₁ Cl ₂ N ₆ OS (MH ⁺ ): 463.0875. Found: 463.0862.

Anal. calculated for C ₂₀ H ₂₀ Cl ₂ N ₆ OS. 3 HCl. 0.5 dioxane: C, 42.84; H, 4.41; Cl, 28.74; N, 13.62; S, 5.20. Found: C, 42.96; H, 4.47; Cl, 28.58; N, 13.53; S, 5.15.

* Example H (3):

(4-Amino-2- (4-piperazin-1-yl-phenylamino) -thiazol-5-yl) - (2,6-dichlorophenyl) -methanone

ΗΝ ^ 'Ά

 r

r r

164 '

The title compound was prepared as described in Example H (1). The title compound

C (105) gave a yellow solid in 44% yield, m.p. 298-300 ° Art.

Ή NMR (DMSO-d ₆ ): δ 7.60-7.50 (5H, m), 7.08 (2H, d, J = 7.8 Hz), 3.44 (4H, bs).

IR (KBr): 3395, 2959, 1618, 1513, 1425 cm &^lt; -1 & gt ^; .

HRFABMS: Calcd for C ₂₀ H ₂₀ Cl ₂ N ₅ OS (MH ⁺ ): 448.0766. Found: 448.0749.

Anal. calculated for C ₂₀ H ₁₉ Cl ₂ N ₅ OS. 1.2 H ₂ O. 0.9 HCl: C, 47.78; H, 4.47; Cl, 20.45; N, 13.93; S, 6.38. Found: C, 47.99; H, 4.38; Cl, 20.57; N, 13.56; S, 6.24.

* Example J (l):

(4-Amino-2- (4-piperazin-1-yl-phenylamino) -thiazol-5-yl) - (2,4,6-trichlorophenyl) -methanone

/4-Amino-2-/4-/4- tert -butoxycarbonyl- piperazin- 1 -yl)-phenylamino/thiazol-5-yl/-(2,2,6- trichlorophenyl).- Methanone having structural

was prepared as described in Example C (1). 1-tert-Butoxycarbonyl-4- (4-isothicyanato-phenyl) -piperazine (from Example C (101) and 2-bromo-2 ', 4', 6'-trichloroacetophenone (from Example C (107)) gave black tar which precipitated from EtOH to give 144 mg (50%) of a yellow amorphous powder, mp 192-193 ° C (d).

Γ

0 c

r 0 rr 0 f 0 0 0 <0 ø 0 r 0 r- ^f '

165 ^X H NMR (DMSO-d₆): δ 7.78 (2H, s), 7.33 (2H, bm), 6.98 (2H, d, J = 9.0 Hz), 3,15-3 1.05 (4H, m), 1.45 (9H, s).

IR (KBr): 3389, 3276, 3166, 1676, 1608, 1577, 1544, 1461, 1421, 1366, 1235, 1202, 1164 cm ^-1 . ·

HRFABMS: Calculated for C ₂₅ H ₂₆ Cl ₃ N ₅ O ₃ SC ₅ (M + Cs ⁺ ): 715.9847. Found: 715.9822.

Anal. calculated for C ₂₅ H ₂₆ Cl ₃ N ₅ O ₃ S. 0.75 H ₂ 0. 0.4 EtOH: C, 50.40; H, 4.90; Cl, 17.30; N, 11.39; S, 5.22. Found: C, 50.69; H, 5.16; Cl, 17.70; N, 10.98; S, 4.90.

The title compound was prepared as follows. (4-Amino-2-4- (4-tert-butoxycarbonyl-piperazin-1-yl) -phenylamino) -thiazol-5-yl] - (2,2,6-trichlorophenyl) -methanone, (50 mg, 0.086 mmol) was stirred in trifluoroacetic acid (TFA;

0.5 ml) at 0 ° C. After 20 min at 0 ° C, a minimum amount of water was added, and saturated NaHCO ₃ was used for neutralization. The obtained suspension was filtered to give a yellow paste which gave a suspension with MeOH / CHCl ₃ and resulted in the isolation of 22 mg (42%) of a yellow amorphous powder.

¹ H NMR (DMSO-d 6): δ 7.80 (2H, s), 7.38 (2H, d, J = 9.0 Hz), 7.01 (2H, d, J = 9.0 Hz).

IR (KBr): 3396, 3284, 3178, 1676, 1614, 1543, 1461, 1423, 1202, 1137 cm ^-1 .

HRFABMS: Calculated for C ₂₀ H ₁₅ C1 ₃ N ₅ OS (MH + ^T): 484.0346. Found:. 484.0333.

Anal. calculated for C ₂₀ H ₁₈ Cl ₃ N ₅ OS. 0.8 MeOH. 0.8 CHC1 _3: C, 42 96; H, 3.67; N, 11.60. Found: C, 42.87; H, 3.45; N, 11.27.

* Example J (2) [4-Amino-2- (4-piperazin-1-yl-phenylamino) -thiazol-5-yl] - (2,6-difluorophenyl) -methanone

P P r P p

166

The title compound was prepared essentially as described in Example J (1). To the title compound of Example C (101) (250 mg, 0.48 mmol) in CH ₂ Cl ₂ at 0 ° C was added TFA (5 mL). After 20 min at 0 ° C, the resulting clear solution was concentrated in vacuo to a residue, which was suspended in a minimum amount of water, cooled to 0 ° C and basified, sat. Na ₂ CO ₃ to pH 9. The solids were combined and recrystallized from EtOH to give 116 mg (58% yield) of a yellow solid, mp 190-193 ° C.

¹ H NMR (DMSO-d ₆ ): δ 8.13 (2H, bs), 7.52 (1H, p, J = 7.3 Hz), 7.36 (2H, d, J = 8.7 Hz) 7.19 (2H, t, J = 8.7 Hz), 6.99 (2H, t, d =

8.7 Hz), 3.24 (4H, bs), 3.13 (4H, bs).

HRFABMS (MH ⁺ ): Calcd .: 416.1357. Found: 416.1370.

Anal. Calcd for C ₂₀ H ₁₉ N ₅ OSF ₂ · 0.7 H ₂ O · 0.7 CF ₃ COOH: C, 49.96; H, 4.11; N, 13.49; S, 6.17.

Found: C, 50.16; H, 4.33; N, 13.14; S, 6.06.

* Example J (3) [4-Amino-2- (4-piperazin-1-yl-phenylamino) -thiazol-5-yl] - (2,4,6-trifluorophenyl) -methanone

NH;

(4-Amino-2-/4-(4- tert.butoxycarbonyl-piperazin- 1 -yl)-phenylamino/thiazol-5-yl/-(2,4,6- trifluorophenyl) methanone, which has a structural formula was prepared essentially as described in Example C (1). r r r »r

167

1-Target butoxycarbonyl-4- (4-isothiocyanato-phenyl) piperazine (from Example 101) and 2'-bromo-2,4,6-trifluoroacetophenone (from Example C (115)) gave a yellow solid that crystallized from EtOH to yield recovery of 200 mg (80%). a yellow amorphous powder that darkened at 125-130 ° C, m.p. 132-135 ° C (dec.).

¹ H NMR (CD ₃ CN): δ 8.69 (1H, bs), 7.46 (2H, d, J = 9.0 Hz),

7.20-7.10 (4H, m), 3.74-3.62 (4H, m), 3.28-3.20 (4H, m), 1.60- (s, 9H).

IR (KBr): 3389, 3282, 3479, 3178, 1686, 1637, 1604, 1546, 1428, 1366, 1343, 1233, 1168, 1121, 1035, 999 cm ^_1.

HRFABMS: Calcd. for C ₂₅ H ₂₇ F ₃ N ₃ O ₃ S (MH): 534.1787. Found: 534.1772.

Anal. calcd for C ₂₅ H ₂₃ N ₅ OSF ₃ .1 H ₂ 0. 0.5 EtOH: C, 54.35; H, 5.44; N, 12.19; S, 5.58.

Found: C, 54.26; H, 5.07; N, 11.92; S, 5,50 ..

The title compound was prepared essentially as described in Example j (1) to give a brown solid which was purified by column chromatography on 10% MeOH / CHCl ₃ as eluent to give 57 mg (60%) of a yellow-orange amorphous solids which decomposed above 205 ° C.

Ή NMR (CD ₃ CN): δ 7.78 (2H, s), 7.42 (2H, d, J = 9.0 Hz), 7.01 (2H, d, J = 9.0 Hz), 3.30-3.18 (4H, m); 3.14-3.02 (4H, m).

IR (KBr): 33406, 1603, 1544, 1430, 1237, 1120, 1034 cm ^-1 .

HRFABMS: Calculated for C ₂₀ H ₁₈ F ₃ N ₅ OS (MH ⁺ ): Calculated: 431.1262. Found: 434.1274.

Anal. calculated for C ₂₀ H ₁₃ N ₅ OSF ₃ · 0.7 MeOH. 0.7 CHC1 _3: C, 47.65; H, 4.02; N, 12.98; S, 5.94.

Found: C, 47.84; H, 3.64; N, 12.59; S, 5.69.

Example J (4):

168

4- (4-Amino-5- (2,6-difluoro-benzoyl) -thiazol-2-ylamino) -N-piperidin-4-ylmethyl-benzenesulfonamide

H!

X

H

N-tert-Butoxycarbonyl-4-carbamoyl-piperidine having the structural formula was prepared as follows. To isonipecotamide (5.00 g, 39.0 mmol) in dioxane (100 mL) was added di-tert-butyl dicarbonate (8.51 g, 39.0 mmol) and N, N-diisopropylethylamine (6.0 mL, 42 mL). , 9 mmol). The mixture was allowed to stir overnight, then evaporated to dryness under reduced pressure. The residue was partitioned between CHCl ₃ and 1N HCl. The organic layer was washed with water and brine, dried over Na ₂ SO ₄ and concentrated to give 8.3 g (93% yield) of a white solid which was used without further purification.

¹ H NMR (CDCl ₃ ): δ 5.53 (2H, bs), 4.03 (2H, d, J = 13.7 Hz), 2.33 (2H, tt, J = 11.8, 3 7 Hz), 2.08 (2H, bs), 1.89 (2H, dd, J =

13.7, 3.7 Hz), 1.69 (1H, dd, J = 11.8, 4.4 Hz), 1.65-1.57 (1H, m), 1.44 (9H, s) ).

4-Aminomethyl-N-tert-butoxycarbonyl-piperidine having the structural formula I was prepared as follows. To N-tert-butoxycarbonyl-4-carbamoyl-piperidine (15.6 mmol) in THF (40 mL) at -78 ° C under Ar was added LiAlH ₄ (592 mg, 15.6 mmol). The mixture was allowed to warm slowly to ambient temperature and after half an hour was again cooled to -78 ° C, quenched with ethyl acetate and partitioned between EtOAc and 2N NaOH. The organic layer was separated, dried over K ₂ CO ₃ and concentrated to give 1.98 g (59% yield) of a yellow slurry which was used without further purification.

<· Γ

169

N-tert-Butoxycarbonyl-4- [(4-nitro-benzenesulfonylamino) -methyl] -piperidine having the structural formula was prepared as follows.

4-Nitrobenzenesulfonyl chloride (2.05 g, 9.24 mmol) was added to the solution of 4-aminomethyl-N-tert. butoxycarbonylpiperidine (1.98 g, 9.24 mmol) in THF (20 mL) at ambient temperature. The mixture was refluxed for 1 hour, concentrated in vacuo and partitioned between CH ₂ Cl ₂ and 1N HCl. The organic layer was washed with brine, dried over Na ₂ SO ₄ , passed through a pad of silica gel and concentrated to give 1.71 g (46%) of a yellow solid which was used without further purification.

4- [(4-Amino-benzenesulfonylamino) -methyl] -N-tert-butoxycarbonyl-piperidine, which has the structural formula was prepared as follows.

N-tert-Butoxycarbonyl-4- [(4-nitro-benzenesulfonylamino) -methyl] -piperidine (1.70 g, 4.26 mmol), 10% Pd / C (250 mg), MeOH (10 mL) and THF (10 mL) was stirred under hydrogen for 2 hours and filtered. The filtrate was concentrated to a residue that was purified by column chromatography with 5% MeOH / CHCl ₃ as eluent to give 1.39 g (88% yield) of a white solid which was used without further purification.

N-tert-Butoxycarbonyl-4- [(4-isothiocyanato-benzenesulfonylamino) -methyl] -piperidine, which has the structural formula • NCS for trans

170 was prepared in a manner analogous to 1- (4-isothiocyanato-phenyl) morpholine for Example C (54). 4 - [(Amino-benzenesulfonylamino) -methyl] - N-tert-butoxycarbonyl-piperidine gave a yellow solid in 39% yield, which was used without further purification.

4- [4- (5-Acetyl-4-amino-thiazol-2-ylamino) -benzenesulfonylamino] -methyl] -N-tert-butoxycarbonyl-piperidine having the structural formula

was prepared in a manner analogous to Example C (1).

N-tert-Butoxycarbonyl-4- [(4-isothiocyanato-benzenesulfonylamino) -methyl] -piperidine and 2-bromo-2 ', 6'-difluoro-acetophenone (from Example C (79)) gave a yellow solid in 50% yield.

¹ H NMR (DMSO-d 6): δ 11.22 (1H, s), 8.20 (2H, bs), 7.84-7.73 (3H, m), 7.62-7.54 (2h) , m), 7.24. (2H, dd, J = 7.8, 7.7 Hz), 3.89 (2H, d, J = 12.8 Hz), 3.35 (2H, s), 2.52 (2H, d) J = 1.2 Hz), 1.60 (2H, d, J = 10.1 Hz), 1.56-1.42 (1H, m), 1.39 (9H, s), 0.91 (2H, d, J = 12.8Hz).

The title compound was prepared in a manner analogous to that used in Example J (1).

4- [4- (5-Acetyl-4-amino-thiazol-2-ylamino) -benzenesulfonylamino] -methyl] -N-tert-butoxycarbonyl-piperidine gave a brown solid in 28% yield.

¹ H NMR (DMSO-d ₆ ): δ 8.11, (2H, bs), 7.70 (4H, bs), 7.58-7.42 (1H, m), 7.20 (1H, d J = 7.8 Hz), 7.15 (1H, d, J = 7.8 Hz), 3.80 (2H, bs), 3.05 (2H, d, J = 10.0 Hz), 2.60 (2H, d, J = 6.8Hz), 1.65 (2H, d, J = 12.2Hz), 1.52 (1H, bs), 1.07 (2H, d, J = 10.0 Hz). HRFABMS (MH ⁺ ): Calcd .: 507.1210. Found: 507.1206.

Anal. calculated for C ₂₂ H ₂₃ N ₅ O ₃ S ₂ F ₂ .0.1 CH ₃ OH. 0.2 CF ₃ COOH: C, 50.65; H, 4.73; N, 13.12; S, 12.02. Found: C, 50.92; H, 4.46; N, rr

171 '· · ·

12.87; S, 12.18.

* Example J (5) [4-Amino-2- / 4- (cis-3,5-dimethyl-piperazin-1-yl) -phenylamino] -N-thiazol-5-yl] - (2,6-difluoro- phenyl) methanone

2,6-cis-dimethyl-4- (4-nitro-phenyl) -piperazine having the structural formula

NO, was prepared essentially as described for the 3R-methyl-1- (4-nitro-phenyl) -piperazine of Example 0 (124). cis-2,6-Dimethylpiperazine gave 2.19 g (100% yield) of a yellow powder, m.p. 130-131.5 ° C, which was used without further purification.

¹ H NMR (CDCl ₃ ): δ 8.03 (2H, d, J = 9.5 Hz), 7.02 (2H, d, J = 9.5 Hz), 3.88 (2H, dd, J = 12.4, 2.0 Hz), 2.82-2.68 (2H, m),

2.44-2.33 (3H, m), 1.03 (6H, d, J = 6.3 Hz).

IR (KBr): 1596, 1509, 1482, 1316, 1252, 1193, 1119, 1101 cm &lt; -1 &gt;

Anal Found for C ₁₂ H ₁₇ N ₃ O ₂ : C, 61.26; H, 7.28; N, 17.86.

Found: C, 61.25; H, 7.42; N, 17.84.

1-tert-Butoxycarbonyl-2,6-dimethyl-4- (4-nitro-phenyl) -piperazine having the structural formula no ₂ was prepared by the following procedure. To 2,6-cis-dimethyl-4- (4-nitrophenyl) -piperazine (1.00 g, 4.25 mmol) in dioxane (20 mL) rree

Di-tert-butyl dicarbonate (1.12 g, 5.12 mmol) and N, N-diisopropylethylamine (1.37 mL, 9.76 mmol) were added. After 3 hours at 80 ° C, the mixture was allowed to cool and evaporate to dryness. The solid was suspended in water, filtered, washed with water and dried under vacuum to give 1.40 g (98% yield) of a yellow powder which was used without further purification.

Δ NMR (CDCl ₃ ): δ 8.12 (2H, ddd, J = 7.3, 2.1, 2.1 Hz), 6.80 (2H, ddd, J = 7.3, 2.1, 2.1 Hz), 4.30 (2H, ddd, J = 13.2, 6.8, 4.5 Hz), 3.71 (d, 2H, J = 13.2 Hz), 3.22 ( dd, 2H, J = 12.8, 4.5 Hz), 1.49 (9H, s), 1.29 (6H, d, J = 6.8 Hz).

IR (KBr): 1689, 1594, 1489, 1400, 1322, 1257, 1057 cm &^lt; -1 & gt ^; .

1- (4-Amino-phenyl) -4-tert-butoxycarbonyl-3,5-dimethyl-piperazine having the structural formula

- ^N - C 'Z - ^NH 2

prepared as follows. Hydrogenation of crude 1-tert-butoxycarbonyl-2,6-dimethyl-4- (4-nitro-phenyl) -piperazine (1.48 g, 4.41 mmol) in THF (20 mL) and MeOH (20 mL) with 10 % Pd / C catalyst gave 1.12 g (83% yield) of a clear sticky oil which was used without further purification.

1 H NMR (CDCl ₃ ): δ 8.13 (2H, d, J = 9.4 Hz), 6.81 (2H, d, J = 9.4 Hz), 4.30 (2H, ddd, J = 13.2, 6.8, 4.5 Hz), 3.71 (2H, d, J = 13.2 Hz), 3.21 (2H, dd, J = 13.2, 4.5 Hz) 1.49 (9H, s), 1.29 (6H, d,

J = 6.8 Hz).

1- (t-Butoxycarbonyl) -2,6-cis-dimethyl-4- (4-isothiocyanato-phenyl) -piperazine having the structural formula

was prepared in a manner analogous to that of 4- (4-isothiocyanatophenyl) -morpholine in Example C (54).

yy yy yy yy yy yy yy yy yyyy yyyyyyyyyyyyy

173 ·

1- (4-Amino-phenyl) -4-tert-butoxycarbonyl-3,5-dimethyl-piperazine afforded a clear sticky foam which was recrystallized from cold ether / hexane to give pale brown crystals in 68% yield, m.p. 97-98 [deg.] C.

¹ H NMR (CDCl 3): δ 6.74 (2H, d, J = 8.7 Hz), 6.67 (2H, d, J = 8.7 Hz), 4.20-4.08 (2H, m), 3.08 (2H, d, J = 11.6 Hz), 2.71 (2H, dd, J = 11.6, 3.9 Hz), 1.41 (9H, s), 1, 28 (6H, d, J = 6.8Hz).

IR (KBr): 2175, 2135, 1691, 1507, 1395, 1341, 1246, 1177, 1098 cm &^lt; -1 & gt ^; .

Anal. Calcd. for C ₁₈ H ₂₅ N ₃ O ₂ S: C, 62.21; H, 7.25; N, 12.09; S, 9.23. Found: C, 62.31; H, 7.32; N, 11.96; S, 9.39.

4-Amino-2- [4- (1-tert-butoxycarbonyl-2,6-dimethyl-piperazin-4-yl) -phenylamino] -thiazol-5-yl- (2,6-difluorophenyl) -methanone having the structural formula

was prepared in a manner analogous to that of Example C (1). 1- (t-Butoxycarbonyl) -2,6-cis-dimethyl-4- (4-isothiocyanato-phenyl) -piperazine and 2-bromo-2 ^' , 6'-difluoroacetophenone (from Example C (79)) gave a yellow solid 51% yield, which was used without further purification.

¹ H NMR (DMSO-d ₆ ): δ 10.66 (1H, s), 8.12 (2H, bs), 7.56-7.44 (1H, m), 7.38 (2H, bs); d, J = 9.0 Hz), 7.18 (1H, d, J = 7.7 Hz), 7.15 (1H, d, J = 8.1 Hz), 6.95 (2H, d, J = 9.0 Hz), 4.14-4.03 (2H, m), 3.49-3.14 (2H, m), 2.75 (2H, dd, J = 12.2, 4.4 Hz) 1.42 (9H, s), 1.24 (6H, d, J = 6.7 Hz).

FABMS (M + Na &lt; + & gt ^; ): 566

The title compound was prepared in a manner analogous to that used in Example J (1).

P P «P e r p r r r r r r p P r

174

4-Amino-2- [4- (1-tert-butoxycarbonyl-2,6-dimethyl-piperazin-4-yl) -phenylamino] -thiazol-5-yl- (2,6-difluorophenyl) -methanone gave a brown powder in 52% yield, mp 293-294.5 ° C.

^{Χ Η} NMR (DMSO-d _6): δ 8.11 (2H, bs), 7.56-7.44 (1H, m), 7.26 (2H, d ,. J = 9.0 Hz) 7.18 (1H, d, J = 7.7Hz), 7.14 (1H, d, J = 8.1Hz), 6.89 (2H, d, J = 9.0Hz), 3, 48 (2H, dd, J = 10.9, 2.2 Hz), 2.88-2.76 (2H, m), 2.07 (4H, t, J = 10.9 Hz), 1.00 (6 H, d, J =

6.3 Hz).

HRFABMS (MH ⁺ ): Calcd .: 444.1670. Found: 444.1658.

Anal. calc. for C ₂₂ H ₂₃ N ₅ OSF ₂ · 0.4 H ₂ O: C, 58.63; H, 5.32; N, 15.54; S, 7.11. Found: C, 58.64; H, 5.40; N, 15.23; S, 6.96.

* Example J (6) [4-Amino-2- / 4- (3,3-dimethyl-piperazin-1-yl) -phenylamino] -thiazol-5-yl] - (2,6-difluoro-phenyl) - methanone

2,2-Dimethyl-4- (4-nitrophenyl) -piperazine, which has the structural formula ^η < ^{ν ν} “Ο- ^ν ° _2, was prepared first by the following procedure. Crude 2,2-dimethylpiperazine (10.0 mmol; Chu et al., Can. J. Chem., Vol. 70 (1992), pp. 1328-1337), 4-fluoro-nitrobenzene (5.0 mmol, 706) mg) and K ₂ CO ₃ (8.3 g, 60.0 mmol) in DMSO (10 mL) was heated at 100 ° C for 4 hours, cooled, diluted with water (100 mL) and extracted with ether: ethyl acetate (200: 50 ml). The organic layer was washed with water (3x) and brine, and concentrated to give 1.17 g (100% yield) of a yellow solid, which was used without further purification.

c r r r

175

Ή NMR (CDCl ₃ ): δ 8.13 (2H, d, J = 9.5 Hz), 6.80 (2H, d, J = 9.5 Hz), 3.38 (2H, dd, J = 5.5, 5.0 Hz), 3.20 (2H, s), 3.07 (2H, dd, J = 5.5, 5.0 Hz), 1.21 (6H, s).

1-tert-Butoxycarbonyl-2,2-dimethyl-4- (4-nitrophenyl) -piperazine having the structural formula was prepared by a method ⁰ analogous to 1- (4-amino-phenyl) -4-tert-butoxycarbonyl) -2 6-dimethyl-piperazine in Example J (5). 2,2-Dimethyl-4- (4-nitrophenyl) -piperazine gave a pale yellow solid in 99% yield which was used without further purification.

¹ H NMR (CDCl ₃ ): δ 8.15 (2H, d, J = 9.4 Hz), 6.64 (2H, d, J = 9.4 Hz), 3.90 (2H, dd, J = 6.0, 5.5 Hz), 3.54 (2H, dd, J = 6.0, 5.5 Hz), 3.53 (2H, s), 1.51 (9H, s), 1 44 (6H, s).

1- (4-Amino-phenyl) -4- (tert-butoxycarbonyl) -3,3-dimethyl-piperazine having the structural formula was prepared as follows. 1-tert-Butoxycarbonyl-2,2-dimethyl-4- (4-nitro-phenyl) -piperazine (700 mg, 2.09 mmol) and 10% Pd / C (100 mg) in THF (15 mL) and MeOH (15 mL) was stirred under hydrogen for 2 hours and filtered. The filtrate was concentrated in vacuo to give a light brown slurry which was used without further purification.

¹ H NMR (CDCl 3): δ 6.69 -5, 65 (4H, m), 3.67 (2H, dd, J = 5.8, 5.4 Hz), 3.21-3.14 (2H m), 3.01 (2H, s), 1.49 (9H, s), 1.43 (6H, s).

1- (t-Butoxycarbonyl) -2,2-dimethyl-4- (4-isothiocyanato-phenyl) -piperazine, which has the structural formula rr

Γ ř

176 was prepared analogously to 4-isothiocyanato-benzamide of C (102). 1- (4-Amino-phenyl) -4-tert. butoxycarbonyl) -3,3-dimethylpiperazine gave a white solid in 80% yield which was used without further purification.

¹ H NMR (CDCl ₃ ): δ 7.15 (2H, d, J = 9.0 Hz), 6.63 (2H, d, J = 9.0 Hz), 3.85 (2H, dd, J = 5.9, 5.5 Hz), 3.42 (2H, dd, J = 5.9, 5.5 Hz), 3.37 (2H, s), 1.57 (9H, s), 1 44 (6H, s).

4-Amino-2- [4- (1-tert-butoxycarbonyl-2,2-dimethyl-piperazin-4-yl) -phenylamino] -thiazol-5-yl- (2,6-difluorophenyl) -methanone, which my structural formula was prepared in a manner analogous to Example C (1).

1- (tert-Butoxycarbonyl) -2,2-dimethyl-4- (4-isothiocyanato-phenyl) -piperazine and 2-bromo-2 ', 6'-difluoro-acetophenone (from Example C (79)) gave a yellow powder in 60% yield, which was used without further purification. '_ ^X H NMR (DMSO-d _6): δ 10.58 (1H, s), 8.13 (2H, bs), 7.61-7.48 (1H, m), 7.40 - 7, 15 (5H, m), 6.79 (2H, d, J = 9.1Hz), 3.74 (2H, dd,

J = 5.8, 5.3 Hz), 3.41-3.30 (4H, m), 1.48 (9H, s), 1.39 (6H, s).

The title compound was prepared in a manner analogous to that used in Example J (1).

4-Amino-2- [4- (1-tert-butoxycarbonyl-2,2-dimethyl-piperazin-4-yl) -phenylamino] -thiazol-5-yl- (2,6-difluorophenyl) -methanone gave yellow solid in 51% yield, mp 205-210 ° C.

¹ H NMR (DMSO-d ₅ ): δ 8.15 (2H, bs), 7.63-7.54 (1H, m), 7.35 (2H, d, J = 9.0 Hz), 7 25 (1H, d, J = 7.7Hz), 7.22 (1H, d, J = 8.1Hz), 6.98 (2H, d, J = 9.0Hz), 3.10 -3.04 (2H, m), 3.02-2, 95 (2H, m,), 2.92 (2H, s), 1.21 (6H, s).

IR (KBr): 3276, 2961, 1620, 1590, 1546, 1516, 1464, 1429, 1364, 1257, 1232, 1002 cm &^lt; -1 & gt ^; .

r r r r ·

<“Rrrrcrr r- r ^r rrr _rrr ^rr · - rrr 'rf - r

177 ·

HRFABMS (MH ⁺ ): Calcd .: 444.1670. Found: 444.1657.

Anal. determined for C ₂₂ H ₂₃ N _with OSF ₂ . 0.7 CH ₃ OH: C, 58.51; H, 5.58; N, 15.03; S, 6.88; Found: C, 58.61; H, 5.68; N, 14.87; S, 6.76.

* Example K:

[4-Amino-2- [4- (4-pyridin-2-yl-piperazin-1-yl) -phenylamino] -thiazol-5-yl] -2,6-difluoro-phenyl) -methanone

N \

1- (4-Nitro-phenyl) -4-pyridin-2-yl-piperazine having the structural formula

0 O ~ ^N ~ 0 ~ _NO2 first prepared in a manner analogous to tert-butyl / methyl (4-nitro-phenyl) amino / -acetate for Example C (103). 1-Pyridin-2-yl-piperazine and 4-fluoronitrobenzene gave a yellow solid in 85% yield.

¹ H NMR (CDCl ₃ ): δ 8.13-8.28 (3H, m), 7.50-7.58 (2H, m), 7.52 (1H, ddd, J = 15.7, 7 3.2 (2.0 Hz), 6.88 (2H, d, J = 9.4 Hz), 6.70 (2H, dd, J = 7.4, 5.1 Hz), 3.78 (4H) δ d, J = 7.4, 5.0 Hz), 3.62 (4H, dd, J = 5.7, 3.3 Hz).

4- (1-Pyridin-2-yl-piperazin-4-yl) aniline having the structural formula.

was prepared in a manner analogous to 4- (4-methyl-piperazin-1-yl) -aniline of Example C (70). 1- (4-Nitro-phenyl) -4-pyridin-2-yl-piperazine gave a gray solid in 94% yield, which was used without further purification.

¹ H NMR (CDCl ₃ ): δ 8.22 (1H, bd, J = 3.5Hz), 7.52 (IH, ddd, J =

178

17.6, 7.2, 1.9 Hz), 6.88 (2H, d, J = 8.7 Hz), 6.66-6.78 (4H, m), 3.72 (4H, dd) J = 5.2, 5.0 Hz), 3.48 (2H, bs), 3.18 (4H, t, J = 5.2, 5.0 Hz).

1- (4-Isothiocyanato-phenyl) -4-pyridin-2-yl-piperazine having the structural formula was prepared in a manner analogous to 1- (4-isothiocyanato-phenyl) -4-pyridin-4-yl-piperazine from Example C (127). 4- (4-Pyridin-2-yl-piperazin-1-yl) -aniline gave 2.2 g (95% yield) of a yellow solid, which was used without further purification.

¹ H NMR (CDCl ₃ ): δ 8.26 (1H, bd, J = 6.3 Hz), 7.91 (1H, ddd, J = 18.1, 7.1, 1.8 Hz), 7 18 (2H, d, J = 9.0 Hz), 6.82-7.00 (4H, m), 4.10 (4H, dd, J = 5.3, 5.1 Hz), 3, 48 (4H, dd, J = 5.3, 5.2 Hz).

The title compound was prepared as follows. To a solution of 1- (4-isothiocyanato-phenyl) -4-pyridin-2-yl-piperazine (25, mg, 0.84 mmol) in dry methanol (4 mL) was added cyanamide (35 mg, 0.84 mmol). and a fresh solution of NaOH (67 mg, 1.67 mmol) in dry MeOH (4 mL). After 1 hour, 2-bromo-2 ', 6'-difluoro-acetophenone (from Example C (79); 178 mg, 0.76 mmol) was added. The next day the resulting yellow suspension was filtered. The solid was washed with H ₂ O and dried under high vacuum to give a yellow solid in 86% yield, mp 138-140 ° C.

¹ H NMR (DMSO-d ₆ ): δ 8.12 (2H, dd, J = 6.5, 1.7 Hz), 7.42-7.60 (2H, m), 7.32 (2H, bd, J = 8.5 Hz), 7.08 (2H, t, J = 9.0 Hz), 6.98 (2H, d, J = 9.0 Hz), 6.88 (2H, d, J = 8.7 Hz), 6.64 (1H, dd, J = 7.0, 5.0 Hz), 3.62 (4H, t, J = 4.7 Hz), 3.20 (4H, t , J = 4.7 Hz).

IR (KBr): 3369, 3180, 2835, 1620, 1597, 1546, 1466, 1433, 1232 cm ^-1 .

HRFABMS: Calcd. for C ₂₅ H ₂₃ F ₂ N ₅ OS (MH ⁺ ): 493.1622. Found:

493.1608.

r r

Γ c

179 Anal. calculated for C ₂₅ H ₂₂ F ₂ N ₆ OS. 0.9 H ₂ O: C, 58.90; H, 4.90; N, 16.49; S, 6.29. Found: C, 58.91; H, 4.64; N, 16.55; S, 6.24.

Example L:

[4-Amino-2- [4- (4-carboxamido-piperidin-1-yl) -phenylamino] -thiazol-5-yl] -2,6-difluorophenyl) -methanone

4-Carboxamido-1- (4-nitro-phenyl) -piperidine having the structural formula No ₂ was prepared in a manner analogous to tert-butyl / methyl- (4-nitrophenyl) -amino] -acetate of Example C (103). 4-Fluoronitrobenzene and isonipecotarnide gave a yellow powder in 98% yield of the raw material which was used without further putification. ¹ H NMR (CD ₃ OD): δ 8.22 (2H, d, J = 9.5 Hz), 7.12 (2H, d, J = 9.5 Hz), 4.20 (2H, d, J = 12.5 Hz), 3.16 (2H, ddd, J = 25.6, 13.3,

2.7 Hz), 2.62-2.70 (1H, m), 2.02 (2H, bd, J = 10.3 Hz), 1.85-1.95 (2H, m).

1- (4-Amino-phenyl) -4-carboxamido-piperidine having the structural formula was prepared in a manner analogous to 4- (4-methyl-piperazin-1-yl) -aniline of Example C (70). 4-Carboxamido-1- (4-nitro-phenyl) -piperidine gave a light yellow powder in 100% yield of the raw material, which was used without further purification.

¹ H NMR (CD 3 OD): δ 6.60 (2H, bs), 6.42 (3H, bs), 3.22 (2H, bs), 2.38 (2H, bs), 2.02 (IH, bs), 1.72-1.92 (4H, m)

Γ *

180

4-Carboxamido-1- (4-isothiocyanatophenyl) -piperidine having the structural formula was prepared in a manner analogous to 1- (4-isothiocyanato-phenyl) -4-pyridin-2-yl-piperazine in Example K (1).

1- (4-Amino-phenyl) -4-carboxamido-piperidine gave a cream colored powder in 93% yield, which was used without further purification.

¹ H NMR (CDCl ₃ ): δ 7.14 (2H, d, J = 9.0 Hz), 6.86 (2H, d, J = 9.0 Hz), 5.50 (1H, bs), 5.30 (1H, bs), 3.74 (2H, d, J = 12.8 Hz), 2.82 (ddd, J = 24.3, 12.5, 2.8 Hz), 2.30 -2.40 (1H, m), 1.80-2.08 (4H, m).

The title compound was prepared as follows. To a solution of 4-carboxamido-1- (4-isothiocyanatophenyl) -piperidine (198 g, 0.76 mmol) in MeOH (3.ml) · was added cyanamide (32 mg, 0.76 mmol) and a solution of sodium methoxide in MeOH (1.65 mL 0.5N, 0.83 mmol). After 30 min, 2-bromo-2 ', 6'-difluoro-acetophenone (162 mg, 0.69 mmol; from Example C (79)) was added. After 2 hours, water was added. The yellow precipitate was filtered off, washed with water, and recrystallized from boiling MeOH to give 200 mg (63% yield) of an amorphous yellow powder, m.p. higher than 300 ° C.

¹ H NMR (DMSO-d ₅ ): δ 7.46-7.58 (1H, m), 7.28 (2H, dd, J = 8.8 Hz)

7.16 (3H, dd, J = 8.0, 7.7 Hz), 6.82 (2H, d, J = 9.1 Hz), 3.68 (2H, bd, J = 12.6 Hz) 3.64 (2H, ddd, J = 13.7, 12.1, 2.8 Hz), 2.04-2.18 (1H, m), 1.52-1.82 (4H, m) .

HRFABMS: Calculated for C ₂₂ H ₂₁ F ₂ N ₅ O ₂ SNa (M + Na ⁺ ): 480.1282. Found: 480.1266.

Anal. determined for C ₂₂ H ₂₁ F ₂ N ₅ O ₂ S. 0.2 H ₂ O: C, 57.31; H, 4.68; N, 15.19; S, 6.95. Found: C, 57.25; H, 4.63; N, 15.31; S, 7.01.

* Example M:

r ©

181

1- (4- / 4-Amino-5- (2,6-difluoro-benzoyl) -thiazol-2-ylamino) -phenyl / piperidine-4-carboxylic acid

HC

1- (4-Nitrophenyl) -piperidine-4-carboxylic acid having the structural formula has been prepared

XXS in a manner analogous to tert-butyl / methyl (4-nitrophenyl) amino / acetate in Example C (103). 4-Fluoronitrobenzene and hexahydropyridine-3-carboxylic acid gave a yellow powder in 89% crude yield, which was used without further purification.

¹ H NMR (CDCl ₃ ): δ 8.00 (2H, d, J = 10.8 Hz), 6.71 (2H, d, J = 10.7 Hz), 3.80 (1H, t, J = 3.9 Hz), 3.72 (1H, t, J = 3.8 Hz), 2.98 (2H, ddd, J = 24.3, 11.0, 3.0 Hz), 2.48 -2.60 (1H, m), 1.88-2.02 (2H, m), 1.68-1.82 (2H, m).

Benzyl 1- (4-nitrophenyl) -piperidine-4-carboxylate having the structural formula was prepared

using the following procedure. Into suspension

1- (4-nitro-phenyl) -piperidine-4-carboxylic acid (500 mg, 2.01 mmol) in acetonitrile (10 mL) was added K ₂ CO ₃ (612 mg, 4.44 mmol) and benzyl bromide ( 265 μΐ, 2.22 ml). The resulting mixture was heated under reflux for 2 hours, allowed to cool and diluted with water. The aqueous layer was extracted with ether (2 x 50 mL).

The combined organic layers were dried over MgSO ₄ , filtered and concentrated in vacuo to give 470 mg (64% crude yield) of a yellow solid which was used without further purification.

¹ H NMR (CDCl ₃ ): 8.13 (2H, d, J = 9.4 Hz), 7.30-7.42 (5H, m), 6.83

182

2.08 (2H, dd, J = 13.5, 3.5 Hz), 1.84-1.94 (2H, m).

. Benzyl 1- (4-aminophenyl) piperidine-4-carboxylate having the structural formula o

was prepared as follows. To a solution of benzyl 1- (4-nitro-phenyl) -piperidine-4-carboxylate (400 mg, 1.18 mmol) in dioxane (5 mL) and ethanol (1 mL) was added stannous chloride (II) dihydrate (1, 2). 06 g, 4.70 mmol). The resulting solution was heated under reflux for 4 hours, allowed to cool, and a small amount of celite was added to the solids aggregation. The mixture was adjusted to pH 8 with saturated aqueous NaHCO ₃ and filtered. The filtrate was diluted with water (50 mL) and extracted with 5% MeOH in CHCl ₃ (2 x 50 mL). The combined organic layers were dried over MgSO ₄ , filtered and concentrated in vacuo to give 400 mg (100% crude yield) of a cream-colored powder which was used without further purification.

^{Σ Η} NMR (CDC1 _3): δ 7.30 (5H, bs), 6.58 (2H, d, J = 8.8 Hz), 6.42 (2H, d, J = 8.8 Hz) 4.94 (2H, s), 3.28 (1H, dd, J = 3.6, 3.1 Hz), 3.18 (1H, dd, J = 3.6, 3.0 Hz), 2 46 (2H, ddd, J = 2.2);

11.8, 2.8 Hz), 2.14-2.28 (1H, m), 1.60-1.88 (4H, m).

Benzyl 1- (4-isothiocyanatophenyl) -piperidine-4-carboxylate having the structural formula was prepared as follows. To a solution of benzyl 1- (4-amino-phenyl) -piperidine-4-carboxylate (400 mg, 1.29 mmol) in THF (5 mL) at -35 ° C was added an excess of Et ₃ N (435 1, 3). , 12 mmol) and thiophosgene (108 L, 1.42 mmol). The resulting mixture was allowed to warm to ambient temperature, stirred for 0.5 h, diluted with water (50 mL) and extracted

E C e e

183 '·

CHCl 3 (2 x 50 mL). The combined organic layers were dried over MgSO ₄ , filtered and concentrated under reduced pressure to give 400 mg (92% yield) of a yellow powder, which was used without further purification.

Ή nm: R (CC > ₃ OD): δ 8,10 (2H, d, J = 9.5Hz) 7.38 (5H, d, J = 4.5) Hz), 6.92 (2H, d, J = 9.5 Hz), 5.18 (2H, s ), 4, 00 (1 H, t, J = 3.4) Hz), 3.96 (1H, dd, J = : 3.5, 3.2 Hz), 3.13 (2H, ddd, J = 24.9 13,8, 2.9 Hz), 2,71-2, 77 (1 H, m), 2.05 (2H , dd, J = 14.1, 3.4 Hz), 1,74- -1.83 (2H, m)

Benzyl 1- [4- (4-amino-5- (2,6-difluorobenzoyl) -thiazol-2-ylamino) -phenyl] -piperidine-4-carboxylate,

The NH 3 having the structural formula was prepared in a manner similar to that described for the title compound of Example C (1). Benzyl 1- (4-isothiocyanato-phenyl) piperidine-4-carboxylate and 2-bromo-2 ', 6'-difluoroacetophenone (from Example C (79)) gave a brown powder in 82% yield, which was used without further purification.

Ή NMR (DMSO-d ₆ ): δ 7.30 (1H, m), 7.18 (2H, d, J = 8.9 Hz), 6.92 (2H, d, J = 9.0 Hz) 4.96 (2H, s), 3.62 (2H, bd, J = 9.2Hz),

2.80 (2H, ddd, J = 26.4, 14.1, 2.6 Hz), 2.36-2.58 (1H, m), 2.04 (2H, bd, J = 3.0 Hz), 1.80-1.92 (2H, m).

The title compound was prepared as follows. To a mixture of benzyl 1- [4- (4-amino-5- (2,6-difluoro-benzoyl) -thiazol-2-yl-amino) -phenyl] -piperidine-4-carboxylate (150 mg, 0.27 mmol) ), in ethanol (10 mL) was added 20% palladium hydroxide on carbon (60 mg). The resulting mixture was stirred under a hydrogen atmosphere for 48 hours. The catalyst was filtered on a pad of celite and washed with ethanol. The filtrate was concentrated under reduced pressure and a minimum amount of í »was added to induce precipitation.

184 · ethyl acetate and CHCl ₃ . The solid was filtered, washed with ethyl acetate and dried to give 40 mg (30%) of a pale blue amorphous powder, mp 275-277 ° C, which was used without further purification.

^X H NMR (DMSO-d _s): δ 8.10 (1H, bs), 7.46-7.58 (1H, m), 7.30 (2H, bd, J = 7.5 Hz), 7 16 (2H, dd, J = 8.0, 7.9 Hz), 6.92 (2H, d, J = 9.1 Hz), 3.58 (2H, bd, J = 12.6 Hz) 2.52 (2ti, dd, J = 11.2, 10.4 Hz), 2.32-2.40 (1H, m), 1.88 (2H, bd, J = 16.1 Hz), 1.58-1.70 (2 H, m).

Anal. calculated for C ₂₂ H ₂₀ F ₂ N ₄ O ₃ S. 0.9 H ₂ O. 0.1 CHC1 _3: C, 54.55; H, 4.54; N, 11.51; S, 6.59. Found: C, 54.55; H, 4.30; N, 11.13;

S, 6.40.

* Example N (l):

(4-Amino-2- (4-nitro-phenylamino) -thiazol-5-yl) - (2-hydroxy-phenyl) -methanone

a * Example N (2):

N- [5- (2-Hydroxy-benzoyl) -2- (4-nitro-phenylamino) -thiazol-4-yl] -benzamide]

Both compounds were obtained from the same experiment. The title compound of Example C (130) was stirred in a mixture of 2.5% aq. KOH (5 eq) in tetrahydrofuran for barley hour. The crude product mixture was separated by flash column chromatography with 5% MeOH / CH ₂ Cl ₂ to give the two title compounds as yellow amorphous substances at 30 and 30% respectively. 50% yield in

Ρ Γ «Γ Γ * · ρ *« ^{'Γ Γ βΓΓΡ} re · ^Γ' ~ (> r <- EFR f- r. R ^r ·

Ρ _Γ Γ. '- r ^Γ . '' r '

185 of Example N (1), respectively. N (2).

For example, N (1): (4-Amino-2- (4-nitro-phenylamino) -thiazol-5-yl) - (2-hydroxy-phenyl) -methanone:

¹ H NMR (DMSO-d ₆ ): δ 11.40 (1H, s), 11.00 (1H, s), 8.24 (4H, d, J =

9.3 Hz), 7.89 (2H, d, J = 9.3 Hz), 7.47 (1H, d, J = 6.9 Hz), 7.34 (1H, dd, J = 7, 9.7 (Hz), 6.92 (2H, d, J = 7.8 Hz).

HRFABMS: Calcd. for C ₁₆ H ₁₂ N ₄ O ₄ S (MH ⁺ ): 357.0658. Found: 357.0660

For Example N (2): N- [5- (2-Hydroxy-benzoyl) -2- (4-nitro-phenylamino) -thiazol-4-yl] -benzamide:

^X H NMR (DMSO-d _s): δ 11.80 (1H, s), 11.60 (1H, s), 10.30 ((4H, d, J

H, s), 8.27 (2H, d, J = 9.2 Hz), 8.00 (2H, d, J = 9.2 Hz), 7.92 (2H, d, J = 7.1 Hz), 7.56-7.68 (3H, m), 7.43 (1H, dd, J = 7.6,

1.6 Hz), 7.34 (1H, ddd, J = 8.5, 7.0, 1.6 Hz), 6.94 (1H, d, J = 8.2 Hz), 6.89 (1H 1 H, dd, J = 7.6, 7.5 Hz).

ESIMS: Calcd. for C ₂₃ H ₁₆ N ₄ O ₅ S (MH ⁺ ): 461. Found: 461.

Other compounds may be prepared according to the invention in a manner similar to those described above. Other exemplary compounds of the invention are identified in Tables I, II and III below, which report the results of biochemical and biological assays.

BIOCHEMICAL AND BIOLOGICAL EVALUATION

Cyclin-dependent kinase activity was measured by quantitation with enzyme-catalyzed, time-dependent incorporation of radioactive phosphate from / ³² P / ATP or / ³³ P / ATP into the protein substrate. Unless otherwise stated, the test was performed at

96-well plates in a total volume of 50 μί in the presence of mM HEPES (N- (2-hydroxyethyl) piperazine-N '- (2-ethanesulfonic acid)) (pH 7.4), 10 mM MgCl ₂ , 25 μΜ adenosine triphosphate ( ATP)

186 '· mg / ml ovalbumin, 5 gg / ml leupeptin, 1 mM dithiothreitol, 10 mM, -glycerophosphate, 0.1 mM sodium vanadate, 1 mM sodium fluoride, 2.5 mM ethylene glycol bis (β-aminoethyl ether) -Ν , Ν, Ν'-N'-tetraacetic acid (EGTA), 2% (v / v) dimethylsulfoxide'a

0.03 - 0.4 pCi / ^32/33 P / ATP per reaction. The reaction was initiated by the enzyme, incubated at 30 ° C and terminated after 20 minutes by adding ethylenediaminetetraacetic acid (EDTA) to 250 mM. Then, the phosphorylated substrate was captured on a nitrocellulose or phosphocellulose membrane using a 96-well multiple filter, and free radioactivity was removed by repeated washing with 85% phosphoric acid. Radioactivity was quantified by exposure of dried phosphor-imaging membranes.

Apparent ICQ values were measured by assaying enzyme activity in the presence of different concentrations of inhibitor compounds and subtracting the background of radioactivity measured in the absence of enzyme. Kinetic parameters (Kcat, Km for ATP) were measured for each enzyme under normal conditions by determining the dependence of baseline rates on ATP concentration. Inhibition data was added to the competitive inhibition equation using Kaleidagraph (Synergy Software), or added to the competitive tightly coupled inhibition equation using KineTic software (BioKin, Ltd).

Inhibition of CDK4 / Cklin D retinoblastoma kinase activity

The complex of human CDK4 and cyclin D3, or the complex of cyclin D1 and the fusion protein of human CDK4 and gluathion-S-transferase (GST-CDK4), or the complex of human CDK4 and genetically engineered (1-264) cyclin D3, was purified using traditional biochemical chromatography methods from insect cells that have been co-infected with the corresponding baculovirus expression vectors (see, eg, Meijer and Kim, Chemical Inhibitors of Cyclin-Dependent Kinases, Methods in Enzymol., Vol. 283 (1997), pp. 113-128).

187

The enzyme complex (5 or 50 nM) was tested with 0.3-0.5 μς of purified recombinant retinoblastoma protein fragment (Rb) as substrate. The engineered Rb fragment (residues 389-928 of native retinoblastoma protein; 62.3 kDa) contains a larger proportion of phosphorylation sites found in the native 106-kDa protein; as well as the end of the six histidine residues to facilitate purification. The phosphorylated Rb substrate was captured by microfiltration on a nitrocellulose membrane and quantified using a phosphor imager as described above. To measure tightly bound inhibitors, the enzyme complex concentration was reduced to 5 nM, and the assay run was extended to 60 minutes, during which time the product-time dependence was linear.

Inhibition of kinase retinoblastoma CDK2 / Cclin A activity

CDK2 was purified using published methodology (Rosenblatt et al., Purification and Crystallization of Human Cyclin-Dependent Kinase 2, J. Mol. Biol., Vol. 230, 1993, pp. 1317-1319) from insect cells infected with baculovirus. expression vector. Cyclone A was purified from E.coli cells expressing full length recombinant cyclin A, and the engineered cyclin A construct was generated by limited proteolysis and purified as previously described. (Jeffrey et al., Mechanism of CDK activation revealed by the structure of the cyclin A-CDK2 complex, Nature, vol. 376 (July 27, 1995), pp. 313-320). Purified, proteolyzed cyclin A was included in the assay with a three-fold five-fold molar excess to CDK2. Alternatively, a complex of CDK2 and proteolyzed cyclin A was prepared and purified by gel filtration. The substrate for this assay was the same Rb substrate fragment used for the CDK4 assays, and the methodology of the CDK2 / cyclin A and CDK4 / cyclin D3 assays were essentially the same except that CDK2 was present at 150 nM or 5 nM. K _t values were measured as described above.

t- r e p r «t p * * <p p! p r p - p r

P P p · 'f

188 · ·

Inhibition of CDK1 (cdc2) / Cvklin B Histon H1 kinase activity:

The complex of human CDK1 (cdc2) and cyclin B was supplied from New England Biolabs (Beverly MA). Alternatively, the CDK1 / glutathione-S-transferase-cyclin B1 complex was purified using glutathione affinity chromatography from insect cells that were co-infected with the corresponding baculovirus expression vectors. Assays were performed as described above at 30 ° C using 2.5 units cdc2 / cyclin B, 10 µg histone H1 protein, and 0.1-0.3 gCi / ^32Z33 P / ATP per assay. The phosphorylated histone substrate was captured by microfiltration on a phosphocellulose P81 membrane and quantified using a phosphor-imager as described above. Κ _χ values were determined using the described curve fitting programs.

Results of tests performed with compounds that include the specific examples described above as well as other examples denoted with the prefix "1" (eg, Examples I (1), 1 (2), etc.) indicating a compound having a known structure (i.e. compound per se known) are listed in Tables I, II and III below. Unless otherwise specified in a specific note, the units and tests used are those indicated in the applicable table column. Abbreviation N.I. indicates that no inhibition was observed at the indicated concentration.

rt rt • · »r <* r f - r

Γ rt r r- Table I ΐς for CDK

Ex. Structure TO, CDK 4 / D (nM) TO, CDK 2 / A (nM) Ki CDK 1 / B (μΜ) KD ’ w H 0 640 '; 102 ^p 460 0.5 1 (2) ’ H 0 8000 '; -30 gM ^b 8700 1 (3) ’ > 5 μΜ “; > 100 μΜ ⁶ A (D 07Φ H 0 660 '; 770 ^p 1200 D (D 490 ^p 900 K4) ’ 3.1 μΜ ¹ ' 4.3 μΜ 3.9 K5) ’ 0 1100 “870 ^p 4600 4.5 C (4) Χψ. 95 " 810 0.09 A (2) HIM -300 " D (2) > 5000 "

and the D-type cyclin is D 3; b = D-type of cyclin is D1; c = D-type of cyclin is limited D3 ν r <· • re t r r r r r * · r - r r

190

Table I K, for CDK (cont.)

Ex. Structure TO,· CDK 4 / D (nM) TO, CDK 2 / A (nM) TO, CDK 1 / B (μΜ) A (3) w 2000 ^a .6100 3.5 C (5) 140 ^a 780 0.293 C (6) H 0 > 10 μΜ ³ 1 (5) ° w Him none @ 10 μΜ " D (3) 2.21 380 ³ 1 (6) fHj - 1500 ³ 2 (8) 4300 ^a K9) 2 # HIM 2500 ³ A (4) 2000 ³ 3100 2.7 1 (9) 22 ' ^H 0 > 20 μΜ ³

a = D-type of cyclin is D 3; b = D-type of cyclin is D1; c = D-type cyclin is restricted to D3

Table I iq for CDK (cont.) * R r * e * e i r c f 'r Γ ** C * i

191 · -

Ex. Structure ~ KT CDK 4 / D (nM) κ, CDK 2 / A (nM) TO, CDK 1 / B (μΜ) 1 (11) H 0 520 " K12) 380 " 4170 1 (13) 2400 " K14) > 50 μΜ " K15) 44- HIM 440 " 1 (16) 1880 " 1 (17) oxo > Η 0 <1000 " K18) ⁰ OJ) > 25 μΜ " K19) P ”Η, Ο- ¹⁴ ►Γ o 1600 " 4800 neX @ 100 ALIGN! μΜ A (5) OCHjCHj 97 " 690 0.163

and the D-type cyclin is D 3; b = D-type of cyclin is D1; c = D-type cyclin is limited by D3 r e e «

192

Table I K for CDK (cont.) R r r f 9 9 'e r 9 f 9 9 9 9 9 9 9 t 9 9 <Γ 9 i r

- f9 r ~ - r '

Ex. Structure TO, CDK 4 / D (nM) TO,· CDK 2 / A (nM) K _from CDK 1 / B (μΜ) E (l) at w H and NOj 420 ’ 320 0.03 A (6) 150 ’ 292 0.052 A (7) 310 ’ A (8) w > 25 μΜ ¹ D (4) H 0 800 ’ A (9) 100 ’ 230 0.053 A (10) W, 36 ’ 318 0.057 1 (20) w H ₀ nent * @ 25 μΜ K21) fH, • H J neru '@ 25 μΜ ’ 1 (22) nenf @ 25 μΜ ’ nenť <§> 100μ M

a = D-type of cyclin is D 3; b = D-type of cyclin is D1; c = D-type cyclin is limited ϋζΚ • β c Γ c

193

Table I K, for CDK (cont.)

Ex. Structure TO, CDK 4 / D (nM) _Row CDK 2 / (nm) TO, CDK 1 / B (μΜ) 1 (23) yjčp or ⁰ 1500 ’ A (ll) 130 ^a 1 (24) Ni 25 μΜ ³ C (7) N 0 510 " C (9) H & 740 ^a C (10) 680 ^a C (8) 27 ^a 389 0.097 C (II) 130 ^a C (12) 27 ^a 670 A (12) 0 9400 ^a 4100

a = D-type of cyclin is D 3; b = D-type of cyclin is D1; c = D-type cyclin is restricted to D3.

♦ e yy yy yy

194

Table I for CDK (cont.) E * r ·· - r r e r © r ©

Ex. Structure TO; CDK 4 / D (nM) _Row CDK 2 / (nm) TO CDK VB (μΜ) C (13) 51 ^a C (14) 57 ^a C (15) H 57 ^a C (16) A + and ti ⁰ 170 ^a C (25) AND 1300 ^a C (24) 8 ^a 248 0.046 C (22) 67 ^a C (17) Wi 72 ^a C (18) Άχα ’ H 0 12900 " nen / @ 10 μΜ C (37) A-Ai. 7 ^a 310 0.233 C (23) W H 330 ^a

a = D-type of cyclin is D 3; b = D-type of cyclin is D1; c = D-type cyclin is limited by D3 f <-

η 195

Table I for CDK (cont.)

Ex. Structure Ki CDK 4 / D (nM) TO,· CDK 2 / A (nM) TO, CDK 1 / B (μΜ) C (19) r-O. xC H 0 15.8 ^a 277 F NH ká Tv H \ CN 0 40.7 ¹ 350 0.2 C (20) ^H ' / Ts CH, * 22 ^a 145 C (21) 117 ^a 480 A (13) £ 250 ^a A (14) Λ 180 ^a H <λ. C (36) NH 3 ^H c, Z 1 CF, 13900 ^a (B) ýt X NI 100 μΜ ¹ C (31) HjCO-A ^ s / q- 5 * H 0 £ Ύ ° Ν.Ϊ. 100 pM ^a C (32) NH, tO, V- 4 H 94 ^a

a = D-type of cyclin is D 3; b = D-type of cyclin is D1; c = D-type cyclin is restricted to D3

C “r f r · <e e e e e * * * * * *

196

Table I Kj. for CDK (cont.)

Ex. Structure Ke CDK 4 / D (nM) TO SDK < 2 / A (nM) TO, SDK 1 / B (pM) C (33) -//AND 57 ^a 20 May C (35) ^H In M ° ll ¹ 23 C (34) .Av . ž H 0 140 ^a 131 C (26) W- 330 ^a C (27) H 1020 ^a C (28) w 240 ^a - C (29) W- 357 ^a C (30) 1400 ^a C (38) 25 ^a 39 C (39) ^HS 0 > 100 pM ^a

a = D-type of cyclin is D 3; b = D-type of cyclin is D1; c = D-type cyclin is restricted to D3

f r

197

Table I ΐς for CDK (cont.)

Ex. Structure TO CDK <4ZD Kf SDK < 2 / A (nM) TO SDK 17B (μΜ) E (2) 1170 ^a C (40) 3840 ^a C (41) 350 ^a 336 C (42) 750 ^a 207 C (43) H ⁰ 315 ^a C (44) - 128 ^a C (45) 30, ¾ <T H 51 ^a 103 0.249 C (46) m δ IN 244 ^a 1790 C (47) H 30 ^a 26 C (48) 14 ^a 11.1 0.015

a = D-type of cyclin is D 3; b - the D-type of cyclin is D1; c = D-type cyclin is limited by D3 e r

198

Table I. for CDK (cont.)

Ex. Structure TO,· CDK 4 / D (nM) TO CDK 2 / A (nM) TO, CDK 1 / B (pM) C (49) HAJ »/“% ⁰ o. ^S 0 23 ^a 10 0.034 C (50) CH, 25.5 ^a 5.6 0.029 C (51) 85 ^a 33.3 C (52) Η H ll ^a 105 C (53) Q-o-Mi, 85 ^a 180 C (54) H ⁰ 34 ^a 453. C (55) 8.5 ^a 493 0.504 C (56) 195 ^a 1020 C (57) | M H CH _a 30 ^a 259 C (58) H ^H I = 34 ^a 306

a = D-type of cyclin is D 3; b = D-type of cyclin is D1; c = D-type cyclin is restricted to D3

199

Table I K, for CDK (cont.)

Ex. Structure TO, CDK 4 / D (nM) TO/ CDK 2 / A (nM) κ, CDK 1 / B (μΜ). C (59) 100 " 135 C (60) 56 " 574 1 (25) 639 " 280 C (61) 120 “ 1200 C (62) 72 " 1710 C (63) 17 " 200 0.133 C (64) 44000 " 42 μΜ C (65) 38000 " 14.3 μΜ C (66) 53 ’ 574 C (67) 3170 " 13.2 μΜ

a = D-type of cyclin is D 3; b = D-type of cyclin is D1; c = D-type cyclin is limited by D3 r 0 r ~ 0

200

Table I for CDK (cont.)

Ex. Structure TO, CDK 4 / D (nM) TO· CDK 2 / A (nM) K _f CDK 1 / B (pM) C (68) > 30 μΜ " C (69) 20 " 638 C (70) 2.8 " 1120 1.37 C (71) H 2 " 482 0.827 C (72) 0 13.5 “ 169 C (73) χ H ^S 0 ^B ' 6.3 " 6.8 0.02 C (74) PMK 4800 " 10.6 μΜ C (75) ooHi 23 " 1080 C (76) H / k / / «I / V ⁰⁴³ H ^U 2000 ' 507 C (77) / '' j / «, rZy *» Hxďr- H 3000 " Nenj @ 5μΜ

a = D-type of cyclin is D 3; b = D-type of cyclin is D1; c = D-type cyclin is restricted to D3

201

Table I K, for CDK (cont.)

Ex. Structure TO,· CDK 4 / D (nM) κ, · SDK 2 / A (nM) Κ · 3DK 1 / B μΜ) C (78) 32 “ 83 C (79) Bye' 54 " 162 C (80) H , 3.3 " 220 0.325 C (81) 12.3 ^a 872 1.24 C (82) y- ⁰ * '. 1 ^M O H 44.2 ^a 467 C (83) 1800 " 32 μΜ C (84) 53 ^a 1040 C (85) f 0 13" 5.7 3.0022 C (86) H 15 " 1100 1.31 C (87) ° o> H 9 " 607 0.826

a = D-type of cyclin is D 3; b = D-type of cyclin is D1; c = D-type cyclin is limited by D3 e r r «

202

Table I K, for CDK (cont.)

Ex. Structure. • Kf CDK 4 / D (nM) _Row CDK 2 / (nm) TO, CDK 1 / B (μΜ) C (88) 7 NHj 444 65 ³ 305 C (89) 55 " 326 G 23 ^a 178 C (90) 2? A 771 C (91) A _B , 6.8 " 81 C (92) ^H A, <PV 63 " 9.4. C (93) H 0 285 “ 1040 C (94) ^{H #} in \ y ° <H 41 " 1040 1.41 C (95) h * 0 ^CHl 25 " <50 0.075 D (5) NH, ’+ Μ H 0 159 " 233

a = D-type of cyclin is D 3; b = D-type of cyclin is D1; c = D-type cyclin is restricted to D3

203

Table I Kj ^ for CDK (cont.)

Ex. Structure TO,· CDK < 4 / D (nM) Ki CDK ( 2 / A (nM) κ · CDK 1 / B (μΜ) C (96) Η 1.5 " 324 1231 H (l) ^{3 HCl} H 8.2 ^a 370 0.681. H (2) ^3HCl T ¹ p H 3.6 ^a 474 0.361 C (97) ⁷ <> ο-Λΰ 8.5 ^a 392 C (98) H 27 ^a 565 0.72 H (3) 2.4 ^a 405 0.472 J (D NH, Η Ό 2.3 ^a 452 0.732 C (99) NH, o 'In the f-O 16.4 ^a 39.5 0.04 C (100) NH TV \ J ^{C? Í} 6.5 ^a 620 0.911 C (101) H 103 ^a 300 0.32

and the D-type cyclin is D 3; b = D-type of cyclin is D1; c = D-type cyclin is restricted to D3

204

Table I K * for CDK (cont.)

Přikl. Structure _Row 4 CDK / D (nM) TO, CDK 2 / A (nM) TO· CDK 1 / B (pM) C (102) 21 ’ 49 0.017 C (103) H 110 " 595 C (104) 190 " 730 C (105) 60 “ 1060 C (106) 134 “ 1460 J (2) CF 3 COOH 6.4 " 135 0.405 C (107) H 'ci 13.8 “ 12.5 C (108) 23 " 6.8 0.009 C (109) 7 * _o 83 " 28 0.035 C (110) 14 " 260 0.104

a = D-type of cyclin is D 3; b = D-type of cyclin is D1; c = D-type cyclin is restricted to D3

205

Table I K for CDK (cont.)

Ex. Structure TO, CDK 4 / D (nM) _Row CDK 2 / (nm) _Row CDK 1 / B (μΜ) C (lll) 21 " 216 C (112) H ₂₃ a 408 C (113) "ME 17.3 " 238 C (114) Η 21 " 8.5 0.028 C (115) O H 57 ^and 55 ^c 18 0.05 J (3) 15 “19 ^c 572 2.0 J (4) 10.2 ^a 13.5 0.022 C (116) NH, 121 " 120 0.077 J (5) 6.3 " 331 0.76 C (117) P p H 10 ^c 423 0.417

a = D-type of cyclin is D 3; b = D-type of cyclin is D1; c = D-type cyclin is limited by D3 rr

206

Table I for CDK (cont.)

Ex. Structure TO, CDK 4 / D (nM) TO, CDK 2 / A (nM) Kt CDK 1 / B (μΜ) C (118) 10.3 " 191 0.097 C (119) H 24 " 86 0.247 C (120) 10.9 " 80 0.062 C (121) 10.6 ' 953 TO H 43 ^c 364 C (122) 35 = 165 J (6) 8.1 ^c 548 0.511 C (123) NH, H 15.4 " 164 C (124) 17 = 611 0.68 C (125) NH, Hj / "P" \ 7 5.4 ^C 602 0.65

a = D-type of cyclin is D 3; b = D-type of cyclin is D1; c = D-type cyclin is restricted to D3

207

Table I for CDK (cont.)

Ex. Structure TO, CDK 4 / D (nM) _Row CDK 2 / (nm) Ke CDK 1 / B (μΜ) L 22 ^c 0.193 M H 46 ^c 290 C (126) /AND 24 ^c 390 C (127) 26 ^c 215 C (128) -0-0 / ¾ 30 ^c 440 C (129) 64 ^c .270

a = D-type of cyclin is D 3; b = D-type of cyclin is D1; c = D-type cyclin is restricted to D3

Inhibition of cell growth: Assessment of cytotoxicity:

Cell growth inhibition was measured using a tetrazolium salt assay based on the ability of living cells to reduce 3- (4,5-dimethylthiazol-2-yl) -2,5- (2H) -diphenyltetrazolium bromide (MTT) to formazan (Mossman (Journal of Immunological Methods, Vol. 65 (1983), pp. 55-58). The water-insoluble violet formazan product was then detected spectrophotometrically. Various cell lines (HCT-116, Saos-2,

208

U2-OS, SW480, COLO-205, RFX-393, M14, MDA-MB-468, and MCF7) were cultured in 96-well plates. Plates were placed in appropriate medium at a volume of 135 ml / well, either in McCoy 5A medium (for Saos-2, U2-OS, SW480 and HCT-116 wells), RPMI (for COLO-205, RFX-393, M14 cells) , or the Minimum Essential Medium Eeagle (for MDA-MB-468 and MCF7 cells). The plates were incubated for 4 hours before the inhibitor compound was added. Various concentrations of inhibitor compounds in 0.5% (v / v) dimethylsulfoxide (15 ml / well) were added, and the cells were incubated at 37 ° C (5% CO ₂ ) for four to six days (depending on cellular). type). At the end of the incubation, MTT was added to a final concentration of 0.2 mg / ml, and the cells were incubated for an additional 4 hours at 37 ° C. After centrifugation of the plates and removal of the medium, the absorbance of formazan (solubilized in dimethylsulfoxide) was measured at 540 nm. The concentration of inhibitor compound exhibiting 50% growth inhibition was determined from the linear fraction of semi-log plotting inhibitor concentration versus percent inhibition. All results were compared to control cells treated with only 0.5% (v / v) dimethylsulfoxide.

* Γ tab.II

209

TABLE II: IC ₅₀ with different cancer cell lines in the MTA assay

with ^ 3, § u> - < with O ) —T AND d H SW-480 C\ rf 0.4,3 15% @ 30 © \ H O @ ,-WITH @ fe 'fO Ό O. £ 00 o in n cf · c4 1.7.3 23% @ 30 5.15 MDA- MB-468 MCF-7 1 m M14 melanoma. COLO- 205 - Saos-2 U2-OS HCT116 2.5,10 O fO Ť O 41% @ 31 in C4 30% @ 30 16% @ 30 26, (85% @) 30) S S G 20, 30 + Γ ⁵ -Sr <S. (I) V WITH- O (e) v C (5) A (4) O? ri HS (ST WH w (9)

• Γ tab.II

210 o

rr vi

fa

O

Tab. II: IC _50, IC ₉₀ (μΜ) of JS

O ε

o p

o υ

CM f

M

O ea

CZ3

C / 0

O cm

P \ o »- (

U ffi

O * 5

V) o

CM

4.10 0.7,7.5 8.3,20.0 0.72,1.7 6.8,15 0.65,5.5 15,> 25 (82%) ». 1.3,4.6 6.2,18.0 2.1,4.8 CO ri rt »—t 14.0> 25 ( 70%) 7.0,18.0 1.0, 1.6 O ro 1 — I · O\ rf 13.0,23.0 8.0,20.0 0.9,1.6 o 'ee ^1-1 ín CM Λ 1.9,6.0 1.9,3.0 .1 16.0,> 25 (80 %) 15.0, > 25 (76%) 1.0,20.0 6.5,> 25 (75%) ) 22.0, > 25 (56%) 12.0,22.0 20.30 0.95,1.8 18> 25 (88%) 2.0,5.0 0.7, 1.5 : 1 10.0,25.0 9.0,23.0 • · 1.0,2.6 6.1, 25 9.0, 22.0 9.0,19.0 E? *, -. < O rH CC at C (12) rr cm at C (37) of-·"" r-l P fa . C (20) C (21) C (35)

r r tab.II

211

Tab. II: IC 50, IC ₉₀ (μΜ)

SW-480 . ⁰⁰ OŘA SI 0.26, 0.60 ! 0.48,1.5 < 0 with 2.6,> 25 (82%) [3.0,> 25 (88%) RXF- 393 0.8, 1.6 > 5.0 M14 melanoma 0.6, 1.8 2.0, 1.8 3 About rS u 0.29, 0.60 1.2,> 5.0 Saos-2 <§ ^Μ Ή rS Λ 2.0,10.0 r— ( in uf ri 0.42,0.75 0 6 rs 0 τ ' 1 1.3,2.8 9.0,22.0 and" ** Os t * · SO 1— (* - ' Whose AND 9.0,20.0 12.0, 26.0 5.8,12.0 4.0,7.0 U2-OS 11.0, > 25 (88%) 5.5,12.0 C 0 rs and O 1.0, 1.5 14.0, 22.0 2.0, 3.0 15.0, 23.0 ts CG ft. ^1-1 'tn rs Λ 16.0, > 26 (84%) 14.0, 26.0 5.0,11.0 0 ss «X en H HCT116 O Os r-t rn TÍ 2.0,8.0 4.2. 7.8 . '_ 1 0.34,0.70 0 T-4 SO <n SO rH rí SO O 8.5,17.0 4.3,17.0 12.0,22.0 5.0,15.0 2.2, 5.9 tí TÍ t> i-5 X 'Si C (38) C (45) rt Tf O C (48) Η— \ Os with 0 C (50) C (52) C (55) z — X SO at Z — X Os SO at C (70) C (71)

tab.II

212

2 5 O es at KM O • n O rt * rt * ^ 2 « ř- SW-480 I ⁰⁰ á š ^§ p rn r-i O? rr O 4.7,9.3 0.22, 0.59 MCF-7 0.4,1.7 3.9,16.0 3.9,13.0 RXF- 393 0.84, 1.3 > 5.0 cq o rt * fO in what τ * cq M14 melanoma 0.92, 2.5 3.8,> 5 0.71,2.2 1.5,4.2 COLO- 205 0.28,1.0 > 5.0 • · 0.24, 0.58 0.39,1.7 Saos-2 1.3, 5.0 VJ r-i C? O* 13.0,22.0 > 25 (15%) 0.28,0.67 and 1 12.22 2.0,4.0 22> 25 (56%) 5.9,23.0 2.2,5.0 1.8,4.5 17.0,25.0 U2-OS in O\ 1-Γ in 1.0, 2.3 O cq rt * eC 25 (1%) 00 ri 1—1. 14, 22 O rr what i-4 6Item no Ό G ΙΛ cq Λ 19.0,> 25 © "C IN eo 3.9,5.8 > 25 HCT116 1.4, 3.4 0.4, 1.1 10.0, 20.0 25 (1%) Ό tn O tn cq © 1 6.5,17.0 0.95, 2.9 19> 25 (70%) _____ 4.7,18.0 1.3, 3.1 1.5,3.0 1.9,5.0 Z ~ \ m ω C (80) C (81) C (82) C (85) (98) C (87)  C (88) z — p C\ '«WZ at O\ AT of- m Os at OF-\ G Q

r r tab.II

213 * r f- ø * f r yr

K r r r Γ i r -. · - r <c r t

Tab. II: IC ₅ o, IC ₉ o (μΜ)

SW-480 MDA- MB-468 - 0.34, 0.7 MCF-7 0.48,1.7 RXF- 393 oo 00 © O 0.55, 1.1 M14 melanoma © Cs * » ττ <S 0.65,2.0 COLO- 205 0.2, 0.6 - • 0.9,2.7 Saos-2 4.2,10.0 ττ J-H rf ί- ο O rš ts © 9.0,20.0 2.7,5.3 | 1.8,3.0 1.0,5.9 1 1.1,2.9 0 * 01‘ST 0.88,2.2 rg r-5 00 u> © O c4 1— » she knows 0.31,3.2 U2-OS 8.0,12.0 0.85,1.5 16.0,21.0 10.0, 21.0 2.4,5.2 3.5,5.8 O\ vi uf ri 4.0,8.0 12.0,22.0 xř she knows i-T ro 1.7,2.8 O ts of 00 xř xř HCT116 m ΓΜ s- \ Λ © ts • 00 00 1— ( 0.58,1.4 and 1 15.0, 25.0 3.9,11.0 1.3, 2.9 0.88,2.4 O ri rn 0.88, 2.7 1 2.3,6.1 i · 1.1, 2.4 0.3, 0.73 2.5, 7.0 0.4, 1.4 cr 'X. C (96) (l) H řř WITH C (97) 00 Os at H (3) Cs AT C (100) C (102) Γ4 C (107) C (108)

tab.II

214 ··

Tab. II: _With the IC, IC ₉₀ (μΜ)

SW-480 MDA- MB-468 0.44, 0.75 MCF-7 0.4, 1.7 to ro g a M14 melanoma COLO- 205 - Saos-2 1.5,14.0 0.89,1.8 3.8,5.9 2.8,9.0 1.3,4.8 1 0.8,2.2 2.2,4.9 0.9, 1.5 0.89,1.5 0.94,1.5 7.0,13.0 2.6,5.2 O es 1-H r-T in 6.0,13.0 3.7,10.0 U2-OS O cs l-H «» Tr 0.7.1.6 3.0,5.5 8.0,19.0 3.0,5.2 2.7,5.0 2.9,5.2 oo 1-H O 1—5 oT O O tZ Tf rs 7.8,12.0 O i-H 1-H in 6.0,11.0 3.5, 6.0 1.5, 4.7 HCT116 1.6,3.0 0.64,1.7 1.7,3.8 3.8, 8.0 1.1, 2.8 0.98,2.1 00 rí C? O SO 1-H O 1.8, 2.9 0.64,1.4 4.0, 7.0 3.5, 6.0 2.5,5.3 3.5, 6.2 1.7, 4.8 C (109) x— \ O ^ H 1-H O x- \ rH i-H 1-H O x-> en PH rH Sex at C (114) řr? 1-H l-H 'w' O x-s en ^ 5 tn C (117) x- \ oo 1-H ΓΗ in-' at X "\ es 1-H 1—5 at X * S O es i-H s ^ X at C (121) C (122)

tab.II

215

^ 3. O C\ at O m υ > -d > —1 pQ C3 H SW-480 MDA- MB-468 1 MCF-7 RXF- 393 M14 melanoma COLO- 205 • Saos-2 0.47, 0.77 2.9,5.7 0.98,1.6 0.9,1.5 he O O 0.51,1.3 7.1,12.0 1.3. 2.5 1 1.2,2.3 HCT116 0.26,0.6 2.7,7.0 0.54,1.6 0.62,1.8 . > 12.5 > 25 0.6, 1.4 $ f · »“ 9 C (123) C (124) C (125) with C (126)

Γ

216 '· · pRb Imunoblotinq:

The ability of compounds to inhibit retinoblast protein (pRb) phosphorylation was tested by Western blot analysis. The anti-Rb antibody was used to measure the conversion of hyper-phosphorylated pRb to hypo-phosphorylated pRb. The anti-phospho-Rb (ser 780) antibody was used to specifically measure dephosphorylation at lilac 780, a site previously shown to be phosphorylated by CDK4 / cyclin D. Inhibition of pRb phosphorylation is indicated by + ”in Table III below, and a defect in inhibition of pRb phosphorylation is indicated in the table.

Human intestinal tumor cells (HCT-116 cells; 5 χ 10 ⁵ ) were seeded in 100 mM dishes and allowed to grow overnight. Five micromoles of each compound was added after 12 hours. The cells were then pooled and centrifuged. Cell pellets were lysed by adding 100 ml lysis buffer (50 mM HEPES (pH 7.0), 250 mM NaCl, 5 mM ethylenediaminetetraacetic acid, 0.1% Nonidet P-40, 1 mM dithiothreitol, 2 mM sodium pyrophosphate, 1 mM orthovanadate sodium, 1 mg / ml aprotin, 1 mg / ml leupeptin, 50 mg / ml phenylmethylsulfonyl fluoride). Forty micrograms of protein were separated by sodium oodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) on a 6% gel. Proteins were transferred to nitrocellulose and blocked overnight with 5% blocking buffer in Tris buffer solution. Anti-Rb antibody (Pharmigen), anti-phospho-Rb (Ser 780) antibody (MBL), and secondary antibody were incubated for one hour at room temperature, followed by three washing steps in 0.01% Twen-20 in Tris-buffer solution. The Rb protein was detected using chemiluminescence according to the manufacturer (Amersham).

r r r *

217 ·

Table III: Inhibition of pRb phosphoxylation

Ex. Merge. It inhibits PRb fosfo- rylaci It inhibits pRb (ser 780) fosfo- rylaci C (85) 4 · + J (2) 4- + C (80) + 4- C (48) 4 · + H (l) 4- 4- C (50) + + C (87) + 4- C (73) 4 · 4- C (81) 4 * + C (94) 4 · + F - - C (71). + - +

The examples above illustrate compounds of Formula I and assays that can be readily performed to determine their activity levels against various CDK / cycles and novel complexes. It is understood that these assays or other suitable assays known in the art may be used to select an inhibitor having the desired level of activity against the selected target.

Although the invention has been illustrated by reference to the specific and patent

In preferred embodiments, one skilled in the art will recognize which variations and modifications can be made by routine experimentation and practice of the invention. For example, one of ordinary skill in the art will determine which variations or substitutions of compounds of Formula I can be prepared without adversely affecting their efficacy in pharmaceutical compositions. Therefore, the present invention is not limited to the foregoing description, but is defined by the appended claims and their equivalents.

Claims (17)

PATENT CLAIMS A compound of formula I wherein: R ¹ is a substituted or unsubstituted group selected from: C _n -alkyl, C ^ g-alkenyl; C 1-6 -alkynyl; C 1-8 -alkoxy; C 1-6 -alcohol; carbocyclic or heterocyclic, monocyclic or fused or unfused polycyclic, cycloalkyl; carbocyclic or heterocyclic, monocyclic or fused or non-fused polycyclic, aryl; carbonyl; ether; (C 1-6 -alkyl) -carbonyl; (C 1-6 -alkyl) -aryl; (C 1-8 -alkyl) -cycloalkyl; (C _1-6 -alkyl) - (C 1-6 -alkoxyyl; aryl- (C 1-6 -alkoxy) thioether, thiol; and sulfonyl; wherein where R ^{1 is} substituted, each substituent is independently halogen; haloalkyl; C 1-6 -alkyl; C 1-8 -alkenyl; C 1-8 -alkynyl; hydroxyl; C 1-6 -altoxyl; amino; nitro; thiol; thioether; imine; cyano; amido; fosfonato; phosphine; carboxyl; thiocarbonyl; sulfonyl; sulfonamide; ketone; aldehyde; an ester; oxygen; carbocyclic or heterocyclic, monocyclic or fused or unfused polycyclic, cycloalkyl; or carbocyclic or heterocyclic, monocyclic or fused or unfused polycyclic, aryl; and R ² is carbocyclic or heterocyclic, monocyclic Or a condensed or unfused polycyclic, ring structure having a substituent at a position adjacent to the point of attachment, wherein the ring structure is optionally further substituted, wherein each R ² is independently halogen; haloalkyl; C 1-6 -alkyl; C 1-6 -alkenyl; C 1-6 -alkynyl; hydroxyl; C 1-6 -alkoxy; amino; nitro; thiol; thioether; imine; cyano; amido; fosfonato; phosphine; carboxyl; thiocarbonyl; sulfonyl; sulfonamide; ketone; aldehyde; an ester; oxygen; carbocyclic or heterocyclic, monocyclic or fused or unfused polycyclic, cycloalkyl; or carbocyclic or heterocyclic, monocyclic or fused or unfused polycyclic, aryl; or a pharmaceutically acceptable salt of a compound of formula I, or a prodrug or pharmaceutically active metabolite of a compound of formula I, or a pharmaceutically acceptable salt thereof. A compound, pharmaceutically acceptable salt, prodrug or pharmaceutically active metabolite thereof according to claim 1, wherein: R ¹ is substituted, each substituent independently halogen; haloalkyl; C 1-8 -alkyl; Cx-g-alkenyl; C 1-6 -alkynyl; hydroxyl; oxygen; C 1-6 -alkoxy; amino; nitro; thiol; thioether; imine; cyano; amido; fosfonato; phosphine; carboxyl; thiocarbonyl; sulfonyl; sulfonamide; ketone; aldehyde; or an ester; and each R ² is independently halogen; haloalkyl; C 1-6 -alkyl; C 1-6 -alkenyl; C 1-6 -alkynyl; hydroxyl; C 1-8 -alkoxy; amino; nitro; thiol; thioether; imine; cyano; amido; fosfonato; phosphine; carboxyl; thiocarbonyl; sulfonyl; sulfonamide; ketone; aldehyde; or an ester. 3. A compound, pharmaceutically acceptable salt, prodrug or compound thereof The active metabolite of claim 1, wherein R ¹ is a substituted phenyl group. The compound, pharmaceutically acceptable salt, prodrug or active metabolite of claim 1, wherein R ¹ is phenyl substituted with an alkylamino or pyridine group. The compound, pharmaceutically acceptable salt, prodrug or active metabolite of claim 1, wherein R ¹ is selected from the group consisting of: e β 222 The compound, pharmaceutically acceptable salt, prodrug or active metabolite of claim 1, wherein R ¹ is phenyl substituted with optionally substituted carbonyl or sulfonamide. The compound, pharmaceutically acceptable salt, prodrug or active metabolite of claim 1, wherein R ¹ is selected from the group consisting of: wherein R ³ is selected from the group consisting of C 1 -C 6 alkyl, C 1 -C 6 alkoxy, aryl, aryloxy and amino. The compound, pharmaceutically acceptable salt, prodrug or active metabolite of claim 1, wherein R ² is ortho-substituted phenyl or thienyl. The compound, pharmaceutically acceptable salt, prodrug or active metabolite of claim 1, wherein R ² is o-halophenyl or o-dihalophenyl. A compound, pharmaceutically acceptable salt, prodrug, or active metabolite of Formula 1, wherein R ² is o-difluorophenyl. e r 223 '· The compound of claim 1 selected from the group consisting of; H ₃ C_ _n ^ .m »h ₃ c -3 HCl NH 4 S 5 S, e and pharmaceutically acceptable metabolites thereof. ^{S HsCk} N ^ kxN. nh ₂ with 's' ^s o H CH ₃ salts, prodrugs and active The compound of claim 1 selected from the group consisting of: •with. 224 and their pharmaceutically acceptable salts, prodrugs, and metabolite activities. 13. A compound selected from the group consisting of: O 225 Or a pharmaceutically acceptable salt, metabolite of said compound. prodrug or active 14. A pharmaceutical composition comprising: (a) an amount of a cell cycle regulating agent, effective to inhibit CDK4 or a CDK4 / cyclin complex, wherein the cell cycle regulating agent is selected from the group consisting of: (I) ««! per r p r r r r R226 (i) compounds of formula I: NH, wherein R ¹ is a substituted or unsubstituted group selected from: C _x . _6- alkyl; C 1-6 -alkenyl; C _x _ ₆ alkynyl; C 1-8 -alkoxy; _ _X C ₆ -alcohol; carbocyclic or heterocyclic, monocyclic or fused or unfused polycyclic, cycloalkyl; carbocyclic or heterocyclic, monocyclic or fused or unfused polycyclic, aryl; carbonyl; ether; (C 1-6 -alkyl) -carbonyl; (C _x. -Al ₆ alkyl) -aryl (C _x _ _s alkyl) -cycloalkyl; (C _1.6 -alkyl) - (C _1.6 -alkoxy) u; aryl- (C _x _G -alkoxy) u; thioether; thiol and sulfonyl; wherein when R ¹ is substituted, each substituent is independently halogen; haloalkyl; C _x _ ₆ -alkyl; C _x _ ₆ alkenyl; C _x . _6- alkynyl; hydroxyl; C _x . _6- alkoxy; amino; nitro; thiol; thioether; imine; cyano; amido; fosfonato; phosphine; carboxyl; thiocarbonyl; sulfonyl; sulfonamide; ketone; aldehyde; an ester; oxygen; carbocyclic or heterocyclic, monocyclic or fused or unfused polycyclic, cycloalkyl; carbocyclic or heterocyclic, monocyclic or fused or unfused polycyclic aryl; and R ² is a carbocyclic or heterocyclic, monocyclic or fused or unfused polycyclic, ring structure having a substituent at a position adjacent to the point of attachment, the ring structure being. optionally further substituted, wherein each R ² is independently a substituent. f · * 227 'halogen; haloalkyl; C 1-8 -alkyl; ? -g-alkenyl; C 1-6 -alkynyl; hydroxyl; C 1-8 -alkoxy; amino; nitro; thiol; thioether; imine cyano; amido; fosfonato; phosphine; carboxyl; thiocarbonyl; sulfonyl; sulfonamide; ketone; aldehyde; an ester; oxygen; carbocyclic or heterocyclic, monocyclic or fused or unfused polycyclic, cycloalkyl; or carbocyclic or heterocyclic, monocyclic or fused or unfused polycyclic, aryl; (ii) a pharmaceutically acceptable salt of a compound of Formula I; and (iii) a prodrug or pharmaceutically active metabolite of a compound of Formula I, or a pharmaceutically acceptable salt thereof; and (b) a pharmaceutically acceptable carrier. A method of treating diseases or disorders mediated by inhibition of CDK4 or CDK4 complex / cyclin comprising administering to a subject in need thereof cell cycle regulating agents selected from the group consisting of: where: R ¹ is a substituted or unsubstituted group selected from: C 1-6 -alkyl; C 1-6 -alkenyl; C 1-6 -alkynyl; C 1-8 -alkoxy; Cx-g-alcohol; carbocyclic or heterocyclic, monocyclic or fused or unfused polycyclic, cycloalkyl; carbocyclic or heterocyclic, monocyclic or 228 condensed or unfused polycyclic, aryl; carbonyl; ether; (C 1-6 -alkyl) -carbonyl; (C 1-6 -alkyl) -aryl (C 1-6 -alkyl) -cycloalkyl; (C 1-6 -alkyl) - (C 1-8 -alkoxy) u; aryl- ( _{C 1-6} -alkoxy) u; thioether; thiol and sulfonyl; wherein when R ¹ is substituted, each substituent is independently halogen; haloalkyl; C 1-6 -alkyl; C 1-6 -alkenyl; C _x . _6- alkynyl; hydroxyl; oxygen; C 1-8 -alkoxy; amino; nitro; thiol; thioether; imine; cyano; amido; fosfonato; phosphine; carboxyl; thiocarbonyl; sulfonyl; sulfonamide; ketone; aldehyde; or an ester; and R ² is a carbocyclic or heterocyclic, monocyclic or fused or unfused polycyclic, ring structure having a substituent at the point of attachment, wherein the ring structure is optionally further substituted wherein each R ² is independently. halogen; haloalkyl; C _x . _6- alkyl; C 1-6 -alkenyl; C 1-6 -alkynyl; hydroxyl; C 1-8 -alkoxy; amino; nitro; thiol; thioether; imine; cyano; amido; fosfonato; phosphine; carboxyl; thiocarbonyl; sulfonyl; sulfonamide; ketone; aldehyde; or an ester; pharmaceutically acceptable salts of compounds of Formula I; and prodrugs and pharmaceutically active metabolites of the compounds of Formula I and pharmaceutically acceptable salts thereof. 16. Compound of Formula I: NH · .O R '(I) e * Ρ Γ r r 0 00, '0 r r ·· 0 Γ - · 229 where: R ¹ is selected from: R ² is substituted or unsubstituted: carbocyclic or heterocyclic, monocyclic or fused or unfused polycyclic, ring structure wherein each optional substituent for R ² is independently halogen; oxygen; haloalkyl ·; C 1-6 -alkyl; C 1-6 -alkenyl; C 1-6 -alkynyl; hydroxyl; C 1-8 -alkoxy; carbocyclic or heterocyclic, monocyclic or fused or unfused polycyclic, cycloalkyl; or carbocyclic or heterocyclic, monocyclic or fused or unfused polycyclic, aryl; amino; nitro; thiol; thioether; imine; cyano; amido; fosfonato; phosphine; carboxyl; thiocarbonyl; sulfonyl; sulfonamide; ketone; aldehyde; or an ester; or a pharmaceutically acceptable salt of a compound of Formula I, or a prodrug or pharmaceutically active metabolite of a compound of Formula I, or a pharmaceutically acceptable salt thereof. 17. A pharmaceutical composition comprising: (a) an effective amount of a cell cycle regulating agent, effective to inhibit CDK or CDK / cyclin complex selected from: er · · ep r _{r r r r r r r r r r r r r r r r r r} r r 230 · (i) compounds of formula I: where: R ¹ is selected from: R ² is substituted or unsubstituted: carbocyclic or heterocyclic, monocyclic or fused or unfused polycyclic, ring structure wherein each optional substituent for R ² is independently, halogen; oxygen; haloalkyl; C 1-8 -alkyl; C 1-8 -alkenyl; C 1-6 -alkynyl; hydroxyl; C 1-8 -alkoxy; carbocyclic or heterocyclic, monocyclic or fused or unfused polycyclic, cycloalkyl; carbocyclic or heterocyclic, monocyclic or fused or unfused polycyclic, aryl; amino; nitro; thiol; thioether; imine; cyano; amido; fosfonato; phosphine; carboxyl; thiocarbonyl; sulfonyl; sulfonamide; ketone; aldehyde; or an ester; 231 '(ii) pharmaceutically acceptable salts of compounds of Formula I; and (iii) prodrugs and pharmaceutically active metabolites of the compounds of Formula I, or pharmaceutically acceptable salts thereof; and (b) a pharmaceutically acceptable carrier.