CN101535295A - Quinoline derivatives for modulating DNA methylation - Google Patents

Abstract

Quinoline derivatives, particularly 4-anilinoquinoline derivatives, are provided. Such quinoline derivatives can be used for modulation of DNA methylation, such as effective inhibition of methylation of cytosine at the C-5 position, for example via selective inhibition of DNA methyltransferase DNMT1. Methods for synthesizing numerous 4-anilinoquinoline derivatives and for modulating DNA methylation are provided. Also provided are methods for formulating and administering these compounds or compositions to treat conditions such as cancer and hematological disorders.

Description

Be used to regulate the quinoline of dna methylation

CROSS-REFERENCE TO RELATED PATENT

The application requires the U.S. Provisional Patent Application No.60/921 of the submission in 12 days October in 2006 under 35 U.S.C. § 119 (e), the U.S. Provisional Patent Application No.60/911 that on April 13rd, 502 and 2007 submitted to, 850 rights and interests, described two provisional application are all incorporated into this paper as a reference in full.

Background of invention

Technical field

The present invention relates to compound, composition, preparation, test kit, usage and the production method of quinoline, more specifically, relate to 4-anilino quinoline, as the conditioning agent of the inhibitor of dna methylation enzyme, dna methylation be used for prevention or the therapeutical agent of treatment and the unusual diseases associated of dna methylation (for example, cancer and haematological malignancies).

Background technology

5 '-m ⁵The C-5 position of the cytosine(Cyt) residue of CpG-3 ' sequence methylates by the silence of gene and play important effect (Smith, A.F.A.Curr.Opin.Genet.Devel., 1999,9,657) in genetic expression.The main enzyme of being responsible for keeping these methylation patterns in reproduction process is dnmt rna DNMT1 (people such as Siedlecki, Biochem.Biophys.Res.Comm., 2003,306,558).Dna methylation is the cause of disease of many heredopathia syndromess, and can have important effect in people's cancer forming process.It is modal molecular changes in the hematopoiesis type vegetation (people such as Egger, Nature, 2004,429,457), and may relate to other tumor type; For example, there is certain schedule of proportion to reveal among the straight cancer patient of sporadic knot to encode the methylating and make this gene silencing (people such as Kane, Cancer Res., 1997,57,808) of gene of MLH1.

The inhibitor DNMT1 that obtains broad research is a for example azacitidine of suicide inhibitor And Decitabine

All be the metabolic antagonist that replaces cytosine(Cyt) to be combined among the DNA and irreversibly catching described enzyme (people such as Egger, Nature, 2004,429,457; People such as Zhou, J.Mol.Biol., 2002,321,599).Two kinds of compounds are used for the treatment of spinal cord heteroplasia syndrome and lymphoproliferative disease now clinically, but sizable toxicity people such as (, Clin.Immunol., 2003,109,89) Leone is all arranged.The chances are owing to incorporated 5-azacytidine in DNA.Decitabine is attached to has hypomethylation (hypomethylation effect) in the DNA chain.Each classification of the cell of differentiation all has the different methylation patterns of oneself.After karyomit(e) repeated, in order to preserve this methylation patterns, the 5-methylcytosine on the fundamental chain was used for directly methylating of complementary daughter DNA chain.5 carbon of cytosine(Cyt) are replaced with nitrogen disturb this normal dna methylation process.Replace 5-methylcytosine at methylated specific position with Decitabine and produce irreversible dnmt rna inactivation, the chances are owing to formed covalent linkage (people such as Juttermann, Proc.Natl.Acad.Sci.USA, 1994 between enzyme and Decitabine, 91,11797).By suppressing DNMT1 (keeping the required enzyme that methylates) specifically, abnormal methylation that can the prophylaxis of tumours suppressor gene.

Described and had only other several micromolecular dna methylation inhibitors, comprise psammaplin sponge metabolite (psammaplin sponge metabolite) (

Deng the people, J.Org.Chem., 2003,68,3866), it is the strong direct inhibitor of dnmt rna, but renders a service less people such as (, Bioorg.Med.Chem.Lett., 2006,16,3330) Godert in test cell line.Other non-nucleoside demethylation reagent for example (-)-epi-nutgall acid catechin-3-gallic acid ester, hydralazine and procainamide also is proved to be and makes effectiveness aspect the gene resurrection than the very different (people such as Chuang of Decitabine, Mol.Cancer Ther., 2005,4,1515).

Therefore, still need to develop and can be used for preventing or treat effective dna methylation conditioning agent with unusual dna methylation diseases associated (for example, cancer and haematological malignancies).

Summary of the invention

In one aspect, the invention provides compound or its physiologically acceptable salt or phosphate precursor medicine or the carboxylicesters or the amino acid ester prodrug of formula (I),

G wherein ₁, G ₂, G ₃, and G ₄Be that C, N or N+ are (at R independently of one another ⁶-R ⁹When being connected in N); G ₅And G ₆Be CH or N independently of one another; G ₇And G ₈Be that CH, C are (at R independently of one another ²When being connected in C), N or N+ be (at R ²When being connected in N),

D ₁And D ₂Be respectively CH, C separately (at R ³When being connected in C), N or N+ be (at R ³When being connected in N).

R ⁶, R ⁷, R ⁸, and R ⁹Be respectively H, halogen, CF separately ₃, OCF ₃, CN, CONHR ⁴, CONR ⁴R ⁵, SO ₂Me, SO ₂NHR ⁴, SO ₂NR ⁴R ⁵, NHCOR ⁴, NHR ⁴, NR ⁴R ⁵, OR ⁴, NO ₂, or CH ₂R ⁴, R wherein ⁴And R ⁵Be H, rudimentary C independently of one another ₁-C ₆Alkyl or cycloalkyl, described rudimentary C ₁-C ₆Alkyl or cycloalkyl is randomly replaced by amino, hydroxyl or methoxy group, one or more Sauerstoffatoms or the nitrogen-atoms part as this cycloalkyl structure is perhaps arranged, this cycloalkyl structure can be represented morpholine, tetramethyleneimine, piperidines, imidazoles or 4-methylpiperazine, perhaps can encircle carbon by-N=replacement-CH=

R ²And R ³Be H, NHR independently of one another ⁴, NR ⁴R ⁵, OR ⁴, NO ₂Or CH ₂R ⁴, R wherein ⁴And R ⁵It is as defined above,

X can be H or the C that randomly replaced by amino, hydroxyl or methoxy group ₁-C ₆Alkyl, or one or more Sauerstoffatoms or the nitrogen-atoms part as the cycloalkyl structure is arranged, it can represent azetidine, tetramethyleneimine, piperidines, piperazine or morpholine;

Y can be CONR ⁴, NR ⁴CO, O, S (O) _n[n=0-2], (CH ₂) _k[k=1-6] ,-CH=CH-, NR ⁴, or two aromatic nucleus between direct connection (that is, the C-C key between two aromatic nucleus), R wherein ⁴And R ⁵As defined above;

O, m and p represent the link position of group Z;

Z can be one of group Q1-Q43 shown in the formula (II);

Wherein A is O or NR ⁴, R wherein ⁴It is as defined above,

G ₉-G ₁₃Be that C, N or N+ are (at R independently of one another ¹⁰-R ¹³When being connected in N); But G ₉-G ₁₃In at least three be C; With

R ₁₀, R ¹¹, R ¹², R ¹³, and R ¹⁴Be respectively H, halogen, alkyl, CF separately ₃, OCF ₃, CN, CONHR ⁴, CONR ⁴R ⁵, SO ₂Me, SO ₂NHR ⁴, SO ₂NR ⁴R ⁵, NHCOR ⁴, NHR ⁴), NR ⁴R ⁵, OR ⁴, NO ₂Or CH ₂R ⁴, R wherein ⁴And R ⁵As defined above.

Should be appreciated that, the compound of formula (I) can be used as different geometry forms and the mapping bodily form exists, the present invention includes the pure form of these independent isomer and its mixture, with and salt derivative or phosphoric acid salt or the carboxylicesters or the amino acid ester prodrug of any neurological progression.

In one aspect of the method, the invention provides the compound of formula (III),

Or its pharmacologically acceptable salt, phosphate precursor medicine or carboxylicesters or amino acid ester prodrug, wherein

R ⁶, R ⁷, R ⁸, and R ⁹Be respectively H, halogen, CF separately ₃, OCF ₃, CN, CONHR ⁴, CONR ⁴R ⁵, SO ₂Me, SO ₂NHR ⁴, SO ₂NR ⁴R ⁵, NHCOR ⁴, NHR ⁴, NR ⁴R ⁵, OR ⁴, NO ₂Or CH ₂R ⁴, R wherein ⁴And R ⁵Be H, C independently of one another ₁-C ₆Alkyl or cycloalkyl, described alkyl and cycloalkyl are randomly replaced by one or more amino, hydroxyl or methoxy group;

X is H or C ₁-C ₆Alkyl, described alkyl are randomly replaced by one or more amino, hydroxyl or methoxy group;

Y is CONR ⁴, NR ⁴CO, O, S (O) _n(n=0-2), (CH ₂) _k(k=1-6) ,-CH=CH-or NR ⁴, R wherein ⁴And R ⁵As defined above;

G ₇And G ₈Be CH or N independently of one another,

D ₁And D ₂Be CH or N independently of one another,

O, m and p represent the link position of group Z;

Z is one of group Q1-Q43 shown in the above-mentioned formula (II), wherein,

A is O or NR ⁴, R wherein ⁴As defined above;

G ₉-G ₁₃Be that C, N or N+ are (at R independently of one another ¹⁰-R ¹³When being connected in N), but G ₉-G ₁₃In at least three be C,

R ¹⁰, R ¹¹, R ¹², R ¹³, and R ¹⁴Be respectively H, halogen, alkyl, CF separately ₃, OCF ₃, CN, CONHR ⁴, CONR ⁴R ⁵, SO ₂Me, SO ₂NHR ⁴, SO ₂NR ⁴R ⁵, NHCOR ⁴, NHR ⁴, NR ⁴R ⁵, OR ⁴, NO ₂Or CH ₂R ⁴, and R wherein ⁴And R ⁵As defined above.

In a further aspect, (for example, cancer) method comprises giving construction (I) or compound or its salt (III) or prodrug, wherein R to the invention provides treatment or prevention and unusual dna methylation diseases associated ²-R ¹⁴, G ₁-G ₁₃, o, m, p, X, Y, Z and A such as above-mentioned respectively for various defined.

Preferably, need treatment or prevent the experimenter of this disease need make its function/activity of keeping methylase DNMT1 that certain reduction is arranged, preferred this function/activity reduces at least 25%, more preferably reduces at least 50%, most preferably reduces at least 75%.Consider that it may be favourable that at least 25% of dna methylation enzymic activity reduces.

Described method also comprises and gives one or more therapeutical agents and/or treatment jointly.Preferred described methods of treatment is included in before the formula (I) or (III) compound or its salt or prodrug that gives above-mentioned definition in addition, the experimenter is given one or more chemotherapeutants or biology therapeutical agent or makes described experimenter experience radiation therapy after the process neutralization.

Although these compounds typically are used for human experimenter's disease prevention or treatment, but they also can be used for the treatment of or prevent other mammiferous disease, warm-blooded animal experimenter (for example, other primate, farming animals for example for example horse, dog and cat of ox and motion animal and pet) for example.

In a further aspect, the invention provides pharmaceutical composition, described pharmaceutical composition comprises formula (I) or compound or its salt (III) or prodrug and the acceptable vehicle of pharmacy, auxiliary, carrier, buffer reagent or the stablizer of above-mentioned definition.

In a further aspect, the invention provides and be used for synthetic or produce the formula (I) of above-mentioned definition or the method for compound or its salt (III) or prodrug.The example of this method is in following reaction scheme 1-6 illustrated.

In a further aspect, the invention provides and produce the preparation method who is used for the medicine of experimenter's administration, described medicine comprises formula (I) or the compound or its salt (III) or the prodrug of above-mentioned definition.

The compound of some preferred formulas (I) is set forth in the following table 1, wherein R ², R ³And R ⁴Be H, and A is NH,

Table 1.

Preferred compounds more of the present invention.

(R ², R ³And R ⁴Be H, D ₁And D ₂For CH, X are that H and A are NH)

R 6-R 9*	Y	Connect	Z	G 5	G 6	G 1	G 2	G 3	G 4	G 7	G 8
												--	CONH	M	Q1	CH	CH	C	C	C	C	CH	CH
--	CONH	m	Q1	N	CH	C	C	C	C	CH	CH
												--	CONH	m	Q1	CH	N	C	C	C	C	CH	CH
--	CONH	m	Q1	CH	CH	N	C	C	C	CH	CH
												--	CONH	m	Q1	CH	CH	C	N	C	C	CH	CH
--	CONH	m	Q1	CH	CH	C	C	N	C	CH	CH
												--	CONH	m	Q1	CH	CH	C	C	C	N	CH	CH
--	CONH	m	Q1	CH	CH	C	C	C	C	N	CH
												--	CONH	m	Q1	CH	CH	C	C	C	C	CH	N
--	CONH	p	Q1	CH	CH	C	C	C	C	CH	CH
												--	CONH	p	Q1	N	CH	C	C	C	C	CH	CH
--	CONH	p	Q1	CH	N	C	C	C	C	CH	CH
												--	CONH	p	Q1	CH	CH	N	C	C	C	CH	CH
--	CONH	p	Q1	CH	CH	C	N	C	C	CH	CH
												--	CONH	p	Q1	CH	CH	C	C	N	C	CH	CH
--	CONH	p	Q1	CH	CH	C	C	C	N	CH	CH
												--	CONH	p	Q1	CH	CH	C	C	C	C	N	CH
--	CONH	p	Q1	CH	CH	C	C	C	C	CH	N
												--	NHCO	m	Q1	CH	CH	C	C	C	C	CH	CH
--	NHCO	p	Q1	CH	CH	C	C	C	C	CH	CH
												--	CONH	p	Q2	CH	CH	C	C	C	C	CH	CH
--	CONH	p	Q2	N	CH	C	C	C	C	CH	CH
												--	CONH	p	Q2	CH	N	C	C	C	C	CH	CH
--	CONH	p	Q2	CH	CH	N	C	C	C	CH	CH

R 6-R 9*	Y	Connect	Z	G 5	G 6	G 1	G 2	G 3	G 4	G 7	G 8
												--	CONH	p	Q2	CH	CH	C	N	C	C	CH	CH
--	CONH	p	Q2	CH	CH	C	C	N	C	CH	CH
												--	CONH	p	Q2	CH	CH	C	C	C	N	CH	CH
--	CONH	p	Q2	CH	CH	C	C	C	C	N	CH
												--	CONH	p	Q2	CH	CH	C	C	C	C	CH	N
--	CONH	p	Q3	CH	CH	C	C	C	C	CH	CH
												--	CONH	p	Q4	CH	CH	C	C	C	C	CH	CH
--	CONH	p	Q4	N	CH	C	C	C	C	CH	CH
												--	CONH	p	Q4	CH	N	C	C	C	C	CH	CH
--	CONH	p	Q4	CH	CH	N	C	C	C	CH	CH
												--	CONH	p	Q4	CH	CH	C	N	C	C	CH	CH
--	CONH	p	Q4	CH	CH	C	C	N	C	CH	CH
												--	CONH	p	Q4	CH	CH	C	C	C	N	CH	CH
--	CONH	p	Q4	CH	CH	C	C	C	C	N	CH
												--	CONH	p	Q4	CH	CH	C	C	C	C	CH	N
--	CONH	m	Q2	CH	CH	C	C	C	C	CH	CH
												--	CONH	m	Q2	N	CH	C	C	C	C	CH	CH
--	CONH	m	Q2	CH	N	C	C	C	C	CH	CH
												--	CONH	m	Q2	CH	CH	N	C	C	C	CH	CH
--	CONH	m	Q2	CH	CH	C	N	C	C	CH	CH
												--	CONH	m	Q2	CH	CH	C	C	N	C	CH	CH
--	CONH	m	Q2	CH	CH	C	C	C	N	CH	CH
												--	CONH	m	Q2	CH	CH	C	C	C	C	N	CH
--	CONH	m	Q2	CH	CH	C	C	C	C	CH	N
												--	CONH	m	Q3	CH	CH	C	C	C	C	CH	CH
--	CONH	m	Q4	CH	CH	C	C	C	C	CH	CH
												--	CONH	m	Q4	N	CH	C	C	C	C	CH	CH
--	CONH	m	Q4	CH	N	C	C	C	C	CH	CH
												--	CONH	m	Q4	CH	CH	N	C	C	C	CH	CH
--	CONH	m	Q4	CH	CH	C	N	C	C	CH	CH
												--	CONH	m	Q4	CH	CH	C	C	N	C	CH	CH
--	CONH	m	Q4	CH	CH	C	C	C	N	CH	CH
												--	CONH	m	Q2	CH	CH	C	C	C	C	N	CH
--	CONH	m	Q2	CH	CH	C	C	C	C	CH	N

R 6-R 9*	Y	Connect	Z	G 5	G 6	G 1	G 2	G 3	G 4	G 7	G 8
												R 7＝NO 2	CONH	m	Q2	CH	CH	C	C	C	C	CH	CH
R 7＝NO 2	CONH	p	Q2	CH	CH	C	C	C	C	CH	CH
												R 7＝NO 2	NHCO	p	Q2	CH	CH	C	C	C	C	CH	CH
R 7＝NH 2	CONH	m	Q1	CH	CH	C	C	C	C	CH	CH
												R 7＝NH 2	CONH	p	Q1	CH	CH	C	C	C	C	CH	CH
R 7＝NH 2	NHCO	m	Q1	CH	CH	C	C	C	C	CH	CH
												R 7＝NH 2	NHCO	p	Q1	CH	CH	C	C	C	C	CH	CH
R 7＝NH 2	CONH	m	Q2	CH	CH	C	C	C	C	CH	CH
												R 7＝NH 2	CONH	p	Q2	CH	CH	C	C	C	C	CH	CH
R 7＝NH 2	NHCO	m	Q2	CH	CH	C	C	C	C	CH	CH
												R 7＝NH 2	NHCO	p	Q2	CH	CH	C	C	C	C	CH	CH
R 7＝NH 2	CONH	m	Q4	CH	CH	C	C	C	C	CH	CH
												R 7＝NH 2	CONH	p	Q4	CH	CH	C	C	C	C	CH	CH
R 7＝NH 2	NHCO	m	Q4	CH	CH	C	C	C	C	CH	CH
												R 7＝NH 2	NHCO	p	Q4	CH	CH	C	C	C	C	CH	CH
R 7＝NH 2	CONH	m	Q2	N	CH	C	C	C	C	CH	CH
												R 7＝NH 2	CONH	p	Q2	N	CH	C	C	C	C	CH	CH
R 7＝NH 2	NHCO	m	Q2	N	CH	C	C	C	C	CH	CH
												R 7＝NH 2	NHCO	p	Q2	N	CH	C	C	C	C	CH	CH
R 7＝NH 2	CONH	m	Q2	CH	N	C	C	C	C	CH	CH
												R 7＝NH 2	CONH	p	Q2	CH	N	C	C	C	C	CH	CH
R 7＝NH 2	NHCO	m	Q2	CH	N	C	C	C	C	CH	CH
												R 7＝NH 2	NHCO	p	Q2	CH	N	C	C	C	C	CH	CH
R 7＝NH 2	CONH	m	Q2	CH	CH	N	C	C	C	CH	CH
												R 7＝NH 2	CONH	p	Q2	CH	CH	N	C	C	C	CH	CH
R 7＝NH 2	NHCO	m	Q2	CH	CH	N	C	C	C	CH	CH
												R 7＝NH 2	NHCO	p	Q2	CH	CH	N	C	C	C	CH	CH
R 7＝NH 2	CONH	m	Q2	CH	CH	C	C	N	C	CH	CH
												R 7＝NH 2	CONH	p	Q2	CH	CH	C	C	N	C	CH	CH
R 7＝NH 2	NHCO	m	Q2	CH	CH	C	C	N	C	CH	CH
												R 7＝NH 2	NHCO	p	Q2	CH	CH	C	C	N	C	CH	CH
R 7＝NH 2	CONH	m	Q2	CH	CH	C	C	C	N	CH	CH
												R 7＝NH 2	CONH	p	Q2	CH	CH	C	C	C	N	CH	CH
R 7＝NH 2	NHCO	m	Q2	CH	CH	C	C	C	N	CH	CH

R 6-R 9*	Y	Connect	Z	G 5	G 6	G 1	G 2	G 3	G 4	G 7	G 8
												R 7＝NH 2	NHCO	p	Q2	CH	CH	C	C	C	N	CH	CH
R 7＝NH 2	CONH	m	Q2	CH	CH	C	C	C	C	N	CH
												R 7＝NH 2	CONH	p	Q2	CH	CH	C	C	C	C	N	CH
R 7＝NH 2	NHCO	m	Q2	CH	CH	C	C	C	C	N	CH
												R 7＝NH 2	NHCO	p	Q2	CH	CH	C	C	C	C	N	CH
R 7＝NH 2	CONH	m	Q2	CH	CH	C	C	C	C	CH	N
												R 7＝NH 2	CONH	p	Q2	CH	CH	C	C	C	C	CH	N
R 7＝NH 2	NHCO	m	Q2	CH	CH	C	C	C	C	CH	N
												R 7＝NH 2	NHCO	p	Q2	CH	CH	C	C	C	C	CH	N
R 7＝NH 2	CONH	m	Q3	CH	CH	C	C	C	C	CH	CH
												R 7＝NH 2	CONH	p	Q3	CH	CH	C	C	C	C	CH	CH
R 7＝NH 2	NHCO	m	Q3	CH	CH	C	C	C	C	CH	CH
												R 7＝NH 2	NHCO	p	Q3	CH	CH	C	C	C	C	CH	CH
R 7＝NMe 2	CONH	p	Q1	CH	CH	C	C	C	C	CH	CH
												R 7＝NMe 2	CONH	m	Q1	CH	CH	C	C	C	C	CH	CH
R 7＝NMe 2	NHCO	p	Q1	CH	CH	C	C	C	C	CH	CH
												R 7＝NMe 2	NHCO	m	Q1	CH	CH	C	C	C	C	CH	CH
R 7＝NMe 2	CONH	p	Q2	CH	CH	C	C	C	C	CH	CH
												R 7＝NMe 2	CONH	m	Q2	CH	CH	C	C	C	C	CH	CH
R 7＝NMe 2	NHCO	p	Q2	CH	CH	C	C	C	C	CH	CH
												R 7＝NMe 2	NHCO	m	Q2	CH	CH	C	C	C	C	CH	CH
R 7＝NMe 2	CONH	p	Q3	CH	CH	C	C	C	C	CH	CH
												R 7＝NMe 2	CONH	m	Q3	CH	CH	C	C	C	C	CH	CH
R 7＝NMe 2	NHCO	p	Q3	CH	CH	C	C	C	C	CH	CH
												R 7＝NMe 2	NHCO	m	Q3	CH	CH	C	C	C	C	CH	CH
R 8＝NO 2	CONH	m	Q1	CH	CH	C	C	C	C	CH	CH
												R 8＝NO 2	CONH	p	Q1	CH	CH	C	C	C	C	CH	CH
R 8＝NO 2	NHCO	m	Q1	CH	CH	C	C	C	C	CH	CH
												R 8＝NO 2	NHCO	p	Q1	CH	CH	C	C	C	C	CH	CH
R 8＝NO 2	CONH	m	Q2	CH	CH	C	C	C	C	CH	CH
												R 8＝NO 2	CONH	p	Q2	CH	CH	C	C	C	C	CH	CH
R 8＝NO 2	NHCO	m	Q2	CH	CH	C	C	C	C	CH	CH
												R 8＝NO 2	NHCO	p	Q2	CH	CH	C	C	C	C	CH	CH
R 8＝NO 2	CONH	m	Q3	CH	CH	C	C	C	C	CH	CH

R 6-R 9*	Y	Connect	Z	G 5	G 6	G 1	G 2	G 3	G 4	G 7	G 8
												R 8＝NO 2	CONH	p	Q3	CH	CH	C	C	C	C	CH	CH
R 8＝NO 2	NHCO	m	Q3	CH	CH	C	C	C	C	CH	CH
												R 8＝NO 2	NHCO	p	Q3	CH	CH	C	C	C	C	CH	CH
R 8＝NH 2	CONH	m	Q1	CH	CH	C	C	C	C	CH	CH
												R 8＝NH 2	CONH	p	Q1	CH	CH	C	C	C	C	CH	CH
R 8＝NH 2	NHCO	m	Q1	CH	CH	C	C	C	C	CH	CH
												R 8＝NH 2	NHCO	p	Q1	CH	CH	C	C	C	C	CH	CH
R 8＝NH 2	CONH	m	Q2	CH	CH	C	C	C	C	CH	CH
												R 8＝NH 2	CONH	p	Q2	CH	CH	C	C	C	C	CH	CH
R 8＝NH 2	NHCO	m	Q2	CH	CH	C	C	C	C	CH	CH
												R 8＝NH 2	NHCO	p	Q2	CH	CH	C	C	C	C	CH	CH
R 8＝NH 2	CONH	m	Q2	N	CH	C	C	C	C	CH	CH
												R 8＝NH 2	CONH	p	Q2	N	CH	C	C	C	C	CH	CH
R 8＝NH 2	NHCO	m	Q2	N	CH	C	C	C	C	CH	CH
												R 8＝NH 2	NHCO	p	Q2	N	CH	C	C	C	C	CH	CH
R 8＝NH 2	CONH	m	Q2	CH	N	C	C	C	C	CH	CH
												R 8＝NH 2	CONH	p	Q2	CH	N	C	C	C	C	CH	CH
R 8＝NH 2	NHCO	m	Q2	CH	N	C	C	C	C	CH	CH
												R 8＝NH 2	NHCO	p	Q2	CH	N	C	C	C	C	CH	CH
R 8＝NH 2	CONH	m	Q2	CH	CH	N	C	C	C	CH	CH
												R 8＝NH 2	CONH	p	Q2	CH	CH	N		C	C	CH	CH
R 8＝NH 2	NHCO	m	Q2	CH	CH	N	C	C	C	CH	CH
												R 8＝NH 2	NHCO	p	Q2	CH	CH	N	C	C	C	CH	CH
R 8＝NH 2	CONH	m	Q2	CH	CH		N		C	CH	CH
												R 8＝NH 2	CONH	p	Q2	CH	CH	C	N	C	C	CH	CH
R 8＝NH 2	NHCO	m	Q2	CH	CH	C	N	C	C	CH	CH
												R 8＝NH 2	NHCO	p	Q2	CH	CH	C	N	C	C	CH	CH
R 8＝NH 2	CONH	m	Q2	CH	CH	C	C	C	N	CH	CH
												R 8＝NH 2	CONH	p	Q2	CH	CH	C	C	C	N	CH	CH
R 8＝NH 2	NHCO	m	Q2	CH	CH	C		C	N	CH	CH
												R 8＝NH 2	NHCO	p	Q2	CH	CH	C	C	C	N	CH	CH
R 8＝NH 2	CONH	m	Q2	CH	CH	C	C	C	C	N	CH
												R 8＝NH 2	CONH	p	Q2	CH	CH	C	C	C	C	N	CH
R 8＝NH 2	NHCO	m	Q2	CH	CH	C	C	C	C	N	CH

R 6-R 9*	Y	Connect	Z	G 5	G 6	G 1	G 2	G 3	G 4	G 7	G 8
												R 8＝NH 2	NHCO	p	Q2	CH	CH	C	C	C	C	N	CH
R 8＝NH 2	CONH	m	Q2	CH	CH	C	C	C	C	CH	N
												R 8＝NH 2	CONH	p	Q2	CH	CH	C	C	C	C	CH	N
R 8＝NH 2	NHCO	m	Q2	CH	CH	C	C	C	C	CH	N
												R 8＝NH 2	NHCO	p	Q2	CH	CH	C	C	C	C	CH	N
R 8＝NH 2	CONH	m	Q2	CH	N	C	C	C	C	CH	N
												R 8＝NH 2	CONH	p	Q2	CH	N	C	C	C	C	CH	N
R 8＝NH 2	NHCO	m	Q2	CH	N	C	C	C	C	CH	N
												R 8＝NH 2	NHCO	p	Q2	CH	N	C	C	C	C	CH	N
R 8＝NH 2	CONH	m	Q3	CH	CH	N	C	C	C	CH	CH
												R 8＝NH 2	CONH	p	Q3	CH	CH	N	C	C	C	CH	CH
R 8＝NH 2	NHCO	m	Q3	CH	CH	N	C	C	C	CH	CH
												R 8＝NH 2	NHCO	p	Q3	CH	CH	N	C	C	C	CH	CH
R 8＝NMe 2	CONH	m	Q1	CH	CH	C	C	C	C	CH	CH
												R 8＝NMe 2	CONH	p	Q1	CH	CH	C	C	C	C	CH	CH
R 8＝NMe 2	CONH	m	Q2	CH	CH	C	C	C	C	CH	CH
												R 8＝NMe 2	CONH	p	Q2	CH	CH	C	C	C	C	CH	CH
R 8＝NMe 2	CONH	m	Q3	CH	CH	C	C	C	C	CH	CH
												R 8＝NMe 2	CONH	p	Q3	CH	CH		C	C	C	CH	CH
R 8＝NMe 2	NHCO	m	Q1	CH	CH	C	C	C	C	CH	CH
												R 8＝NMe 2	NHCO	p	Q1	CH	CH	C	C	C	C	CH	CH
R 8＝NMe 2	NHCO	m	Q2	CH	CH	C	C	C	C	CH	CH
												R 8＝NMe 2	NHCO	p	Q2	CH	CH	C	C	C	C	CH	CH
R 8＝NMe 2	NHCO	m	Q3	CH	CH	C	C	C	C	CH	CH
												R 8＝NMe 2	NHCO	p	Q3	CH	CH	C	C	C	C	CH	CH

If * do not indicate, R then ⁶-R ⁹=H

Recognize that some compound of the present invention can exist with one or more different enantiomorphs or diastereomer form.Should be appreciated that described enantiomorph or diastereomer form all are included in the above-mentioned all respects of the present invention.

The term " the acceptable salt of pharmacology " that uses from start to finish in specification sheets is construed as and is meant any acid or alkali deutero-salt, described salt is by hydrochloric acid, sulfuric acid, phosphoric acid, acetate, citric acid, oxalic acid, propanedioic acid, Whitfield's ointment, oxysuccinic acid, FUMARIC ACID TECH GRADE, succsinic acid, xitix, toxilic acid, methanesulfonic, hydroxyethylsulfonic acid (isoethonic acid) etc., and formation such as salt of wormwood, sodium hydroxide or potassium hydroxide, ammoniacal liquor, triethylamine, trolamine.

In some embodiments of the present invention, the salt of formula (I) or compound (III) is to form with following acid: hydrochloric acid, Hydrogen bromide, sulfuric acid, phosphoric acid, carboxylic acid, sulfonic acid, thioic acid sulfoacid (sulfoacid) or phosphonic acids (phospho acid), acetate, propionic acid, oxyacetic acid, Succinic Acid, toxilic acid, hydroxymaleic acid, methyl-maleic acid, FUMARIC ACID TECH GRADE, oxysuccinic acid, tartrate, lactic acid, oxalic acid, glyconic acid, saccharic acid, glucuronic acid, citric acid, phenylformic acid, styracin, amygdalic acid, Whitfield's ointment, the 4-aminosallcylic acid, the 2-phenoxy benzoic acid, the 2-acetoxy-benzoic acid, pamoic acid, nicotinic acid, Yi Yansuan, amino acid, L-glutamic acid, Aspartic Acid, phenylacetic acid, methylsulfonic acid, ethyl sulfonic acid, the 2-hydroxyethanesulfonic acid, ethane-1, the 2-disulfonic acid, Phenylsulfonic acid, the 4-toluene sulfonic acide, naphthalene-2-sulfonic acid, naphthalene-1, the 5-disulfonic acid, 2-or 3-phoshoglyceric acid, Robison ester, N-cyclohexyl thionamic acid, and xitix.In some embodiments, described salt is sodium salt, calcium salt, lithium salts, sylvite, ammonium salt or trialkyl ammonium salts.

In some preferred embodiments, Z is Q1, Q2, Q3, Q4, Q9 or Q10.In some preferred embodiments, described compound is the structure of formula (III), R ⁶, R ⁷, R ⁸, and R ⁹H respectively does for oneself; X is H; Y is CONH or NHCO; Z is Q2 or Q9; A is that NH and Z are connected m or p position.In some preferred embodiments, described compound is the structure of formula (III), R ⁶, R ⁷, R ⁸, and R ⁹H respectively does for oneself; X is H; Y is CONH, and Z is Q2; A is NH; Be connected the p position with Z.In some preferred embodiments, described compound is the structure of formula (III), R ⁶, R ⁷, R ⁸, and R ⁹H respectively does for oneself; X is H; Y is CONH; Z is Q9; A is NH; Be connected the p position with Z.In some preferred embodiments, described compound is the structure of formula (III), R ⁶, R ⁷, R ⁸, and R ⁹H respectively does for oneself; X is H; Y is CONH or NHCO; Z is Q2; A is that NH and Z are connected the m position.In some preferred embodiments, described compound is the structure of formula (III), R ⁶, R ⁸, and R ⁹H respectively does for oneself; R ⁷Be NMe ₂X is H; Y is CONH or NHCO; Z is Q2; A is that NH and Z are connected the m position.In some preferred embodiments, described compound is the structure of formula (III), R ⁶, R ⁸, and R ⁹H respectively does for oneself; R ⁷Be Cl; X is H; Y is CONH; Z is Q2; A is that NH and Z are connected the p position.In some preferred embodiments, described compound is the structure of formula (III), R ⁶, R ⁷, R ⁸, and R ⁹Be respectively H, F or Cl.

One aspect of the invention is pharmaceutical composition, described pharmaceutical composition comprises formula (I) or compound (III), salt or the prodrug of above-mentioned definition, and pharmaceutically acceptable carrier.In some embodiments of described pharmaceutical composition, described compound, salt or prodrug are solid form.In some embodiments, described pharmaceutical composition is an oral dosage form.In some embodiments, described pharmaceutical composition is injectable formulation.In some embodiments, described pharmaceutical composition is a topical dosage forms.

One aspect of the invention is the method that suppresses the dna methylation in the cell, comprising: make formula (I) or the compound or its salt (III) or the prodrug of the above-mentioned definition of cells contacting, make that the dna methylation activity in the cell is suppressed.

One aspect of the invention is the method that suppresses dna methylation in the cell, comprising: make that cells contacting is above-mentioned to be defined in formula (I) or (III) at compound or its salt or prodrug, to make that the dnmt rna activity in the cell is suppressed.In some embodiments, the activity that suppresses dnmt rna by the degraded of dnmt rna DNMT1.In some embodiments, described contact procedure comprises formula (I) or the compound or its salt (III) or the prodrug of the above-mentioned definition that makes cells contacting biology significant quantity, makes the activity inhibited at least 50% of dnmt rna DNMT1 in the cell.In some embodiments, described contact procedure comprises formula (I) or the compound or its salt (III) or the prodrug of the above-mentioned definition that makes cells contacting biology significant quantity, makes the activity inhibited at least 25% of dnmt rna DNMT1 in the cell.

One aspect of the invention is the method for the activation recovering that is used for making the repressed gene of cell dna methylation, comprise: make formula (I) or the compound or its salt (III) or the prodrug of the above-mentioned definition of cells contacting biology significant quantity, make the activity of the repressed gene of dna methylation raise at least 25% with respect to the situation under the existence that does not have described compound, salt or prodrug.In some embodiments, the step of contact comprises formula (I) or the compound or its salt (III) or the prodrug of the above-mentioned definition that makes cells contacting biology significant quantity, makes the methylate transcriptional activity or the transcriptional level of repressed gene transcription thing of DNA-raise at least 25%.In some embodiments, the repressed gene of described dna methylation is selected from following group: 14-3-3Sigma, ABL1 (P1), ABO, APC, AR (androgen receptor), BLT1 (leukotrienes B4 acceptor), BRCA1, CALCA (thyrocalcitonin), CASP8 (caspase 8), caveolin 1, CD44, CFTR, COX2, CSPG2 (versican), CX26 (connection protein 26), Cyclin (cyclin) A1, DBCCR1, ECAD (E-cadherin), endothelin-receptor B, EPHA3, EPO (erythropoietin), ER (estrogen receptor), FHIT, GPC3 (glypican 3), GST-pi, H19, H-cadherin (CDH13), gamma globulin, HIC1, hMLH1, HOXA5, IGF2 (insulin-like growth factor II), IGFBP7, IRF7, LKB1, LRP-2 (huge albumen), MDGI (growth inhibitor in breast source), MDR1, MDR3 (PGY3), MGMT (06 methyl guanine methyl transferase), MUC2, MYOD1, N33, NEP (neutral endopeptidase 24.1)/CALLA, NIS (sodium iodide symport gene), P14/ARF, P15 (CDKN2B), P16 (CDKN2A), P27KIP1, p57KIP2, PAX6, PgR (PgR), RAR-β 2, RASSF1, RB1 (retinoblastoma), TERT, TESTIN, TGFBRI, THBS1 (thrombospondin-1), TIMP3, TLS3 (T-plastin), urokinase (uPA), VHL (brain retinal blood tuberculation), WT1, and ZO2 (zonuls occludens 2).The network address of M.D.ANderson Cancer Center provides the details about these tumor suppressor genes.Referring to www.mdanderson.org/departments/methylation/dIndex.cfm? the network address of pn=D02B3250-57D7-4F61-88358636A8073A08, it is incorporated into this paper as a reference in full.

One aspect of the invention is and be used for the treatment of the method for suffering from the patient of unusual dna methylation diseases associated, comprise: give pharmaceutical composition to described patient, described pharmaceutical composition comprises formula (I) or compound or its salt (III) or the prodrug and the pharmaceutically acceptable carrier of the above-mentioned definition for the treatment of significant quantity.In some embodiments, described composition is by following administration: oral administration, parenterai administration, topical, intraperitoneal administration, intravenous administration, intra-arterial administration, transdermal administration, sublingual administration, intramuscular administration, rectal administration, transbuccally administration, intranasal administration, liposome administration, by inhalation, vagina administration, eye drops, by administration (intraadiposally) in local delivery administration, subcutaneous administration, the fat, intra-articular administration or intrathecal drug delivery.In some embodiments, described pharmaceutical composition is an oral administration.In some embodiments, described method comprises in addition and gives second therapeutical agent with described pharmaceutical composition to described patient.In some embodiments, described second therapeutical agent is Decitabine or azacitidine.In some embodiments, described second therapeutical agent is selected from medicine, biology medicine, interleukin-, Interferon, rabbit, cytokine, immunomodulator and the monoclonal antibody of histone deacylase inhibitor, antibiolics, alkylating agent, retinoid, hormone drug, plant origin.In some embodiments, described histone deacylase inhibitor is selected from the two hydroxamic acid of bent ancient rhzomorph A, cork anilid hydroxamic acid, oxamflatin, cork acid, the two hydroxamic acid of a carboxylic acid styracin, pyroxamide, trapoxin A, apicidin, depsipeptide, N-(2-aminophenyl)-4-[N-(pyridin-3-yl methoxycarbonyl) amino methyl] benzamide, butyric acid, phenyl butyrate and arginine butyric ester.

In some embodiments, described and unusual dna methylation diseases associated is selected from hematology illness, innocent tumour and cancer.In some embodiments, described hematology illness is selected from acute myelogenous leukemia, acute promyelocytic leukemia, acute lymphoblastic leukemia, chronic graininess leukemia, spinal cord heteroplasia syndrome and sickle cell disease.In some embodiments, described disease is a cancer, and is selected from following: mammary cancer, skin carcinoma, osteocarcinoma, prostate cancer, liver cancer, lung cancer, nonsmall-cell lung cancer, the cancer of the brain, laryngocarcinoma, carcinoma of gallbladder, carcinoma of the pancreas, the rectum cancer, parathyroid carcinoma, thyroid carcinoma, adrenal carcinoma, nervous center is organized cancer, head and neck cancer, colorectal carcinoma, cancer of the stomach, bronchogenic carcinoma, and kidney, rodent cancer, ulcer type and nipple type squamous cell carcinoma, the transitivity skin carcinoma, osteosarcoma, Ewing sarcoma, non-Hodgkin lymphomas (veticulum cell sarcoma), myelomatosis, giant cell tumor, small cell lung tumor, gallbladdergallstonecholetithiasis, islet cell tumor, primary brain tumors, acute and chronic lymphocytic knurl and granulocyte knurl, the hair cell knurl, adenoma, hyperplasia, medullary carcinoma, pheochromocytoma, mucous membrane neuroma (mucosal neuronms), enteron aisle gangliocytoma (intestinal ganglloneuromas), hyperplasia corneal nerve knurl, the marfanoidhabitus knurl, Wilm ' s knurl, spermocytoma, ovarian tumor, leiomyoma, cervical atypism hyperplasia and carcinoma in situ, neuroblastoma, retinoblastoma, soft tissue sarcoma, the carcinoid malignant knurl, partial skin lesion, cutaneous T cell lymphoma (mycosis fungoide), rhabdosarcoma, Kaposi, osteogenic sarcoma, pernicious hypercalcemia, the nephrocyte tumour, polycythemia vera (polycythermia vera), gland cancer, glioblastoma multiforme (glioblastoma multiforma), leukemia, lymphoma, malignant melanoma, and epidermoid carcinoma.

In a further aspect, the invention provides the method for the dna methylation that is used for suppressing cell, comprise: the formula (I) or compound or its salt (III) or the prodrug that make the above-mentioned definition of cells contacting, make the dna methylation activity in the cell be inhibited with respect to giving compound dna methylation activity before, preferably be suppressed 25%, more preferably preferably be suppressed 50%.

In a further aspect, the invention provides and be used for the active method of dnmt rna DNMT1 that selectivity suppresses cell, comprise: make cells contacting compound of the present invention, make the degree of activity inhibited of the dnmt rna DNMT1 in the cell greater than dnmt rna DNMT3a or the repressed degree of DNMT3b.

According to described method, the activity of dnmt rna DNMT1 is inhibited by the degraded of dnmt rna DNMT1.

According to described method, the step of contact comprises the compound of the present invention that makes cells contacting biology significant quantity, makes dnmt rna DNMT1 activity inhibited at least 50% in the cell.

According to described method, the step of contact comprises the compound of the present invention that makes cells contacting biology significant quantity, makes dnmt rna DNMT1 activity inhibited at least 25% in the cell.

In a further aspect, the invention provides the active method of the repressed gene of dna methylation that is used for recovering cell, comprise: the compound of the present invention that makes cells contacting biology significant quantity, make the activity of the repressed gene of dna methylation raise at least 25%, preferred rising at least 50% with respect to the situation that does not have compound to exist.The gene transcription activity that the activity of the repressed gene of dna methylation includes but not limited to methylate and is suppressed.

In a further aspect, the invention provides and be used for the treatment of the method for suffering from the patient of unusual dna methylation diseases associated, comprise: give pharmaceutical composition to described patient, described pharmaceutical composition comprises formula (I) or compound or its salt (III) or the prodrug and the pharmaceutically acceptable carrier of the above-mentioned definition for the treatment of significant quantity.

According to described method, described composition is by following administration: oral administration, parenterai administration, topical, intraperitoneal administration, intravenous administration, intra-arterial administration, transdermal administration, sublingual administration, intramuscular administration, rectal administration, transbuccally administration, intranasal administration, liposome administration, by inhalation, vagina administration, eye drops, by administration in local delivery administration, subcutaneous administration, the fat, intra-articular administration or intrathecal drug delivery.

According to described method, described method comprises in addition: give second therapeutical agent with described pharmaceutical composition to the patient.

According to described method, described second therapeutical agent can be Decitabine or azacitidine.

According to described method, described second therapeutical agent can be selected from medicament, biology medicament, interleukin-, Interferon, rabbit, cytokine, immunomodulator and the monoclonal antibody of histone deacylase inhibitor, antibiolics, alkylating agent, retinoid, hormone drug, plant origin.

According to described method, described and unusual dna methylation diseases associated is selected from hematology illness, innocent tumour and cancer.

The example of cancer includes but not limited to acute myelogenous leukemia, acute promyelocytic leukemia, acute lymphoblastic leukemia, chronic graininess leukemia, spinal cord heteroplasia syndrome and sickle cell disease.

The example of hematology illness includes but not limited to mammary cancer, skin carcinoma, osteocarcinoma, prostate cancer, liver cancer, lung cancer, nonsmall-cell lung cancer, the cancer of the brain, laryngocarcinoma, carcinoma of gallbladder, carcinoma of the pancreas, the rectum cancer, parathyroid carcinoma, thyroid carcinoma, adrenal carcinoma, nervous center is organized cancer, head and neck cancer, colorectal carcinoma, cancer of the stomach, bronchogenic carcinoma, and kidney, rodent cancer, ulcer type and nipple type squamous cell carcinoma, the transitivity skin carcinoma, osteosarcoma, Ewing sarcoma, non-Hodgkin lymphomas (veticulum cell sarcoma), myelomatosis, giant cell tumor, small cell lung tumor, gallbladdergallstonecholetithiasis, islet cell tumor, primary brain tumors, acute and chronic lymphocytic knurl and granulocyte knurl, the hair cell knurl, adenoma, hyperplasia, medullary carcinoma, pheochromocytoma, mucous membrane neuroma (mucosal neuronms), enteron aisle gangliocytoma (intestinalganglloneuromas), hyperplasia corneal nerve knurl, marfanoid habitus knurl, Wilm ' s knurl, spermocytoma, ovarian tumor, leiomyoma, cervical atypism hyperplasia and carcinoma in situ, neuroblastoma, retinoblastoma, soft tissue sarcoma, the carcinoid malignant knurl, partial skin lesion, cutaneous T cell lymphoma (mycosis fungoide), rhabdosarcoma, Kaposi, osteogenic sarcoma, pernicious hypercalcinemia, the nephrocyte tumour, polycythemia vera (polycythermia vera), gland cancer, glioblastoma multiforme (glioblastoma multiforma), leukemia, lymphoma, malignant melanoma, and epidermoid carcinoma.

The other aspect of the present invention can become apparent from following description, below describes just providing as an example, and with reference to the building-up reactions scheme of enclosing.

Incorporating into of reference

All publications and the patent application mentioned at this specification sheets all are merged in this paper as a reference, are equivalent to show clearly and respectively the degree that publication that each is independent and patent application all are incorporated by reference.

The accompanying drawing summary

Fig. 1 illustrates the rising of the rna expression level of the p16 that the representative compounds of the present invention by different concns causes.

Detailed Description Of The Invention

Although showed and described the preferred embodiments of the invention, it will be apparent to one skilled in the art that this embodiment just provides by example herein. Can expect many variants, variation and replacement and not deviate from the present invention by those skilled in the art. Should be appreciated that the various replacement schemes of described embodiment of the present invention may be used to put into practice the present invention herein. The claim that is intended to subsequently defines scope of the present invention, and contains thus method and structure and equivalent thereof in these claim scopes.

The invention provides compound, composition, preparation, kit, usage and the production method of quinoline, more specifically, relate to 4-anilino-quinoline, therapeutic agent as the conditioning agent of the agent of dna methylation enzymeinhibition, the dna methylation disease (for example, cancer and haematological malignancies) unusually relevant with dna methylation with being used for prevention or treatment.

We have found unexpectedly that a series of 4-anilino-quinoline compounds (for example, the compound of formula (I) or formula (III)) suppress the cell function of DNA transmethylase DNMT1 specifically by optionally degrading DNMT1. By contrast, the two quaternary salts of former 4-anilino-quinoline have shown and have been incorporated into DNA ditch (people such as Leupin, Biochemistry, 1986,25,5902; The people such as Squire, Nucleic Acids Res., 1997,25,4072). These compounds have shown as cytotoxic, and develop as anticarcinogen at first. (Atwell ﹠ Cain, J.Med.Chem., 1973,16,673; The people such as Denny, J. Med.Chem., 1979,22,134), but example (NSC 176319) toxicity of finding to be considered for clinical testing too large (Plowman ﹠ Adamson, Pharmacol., 1978,17,61).

Although do not wish to be bound by the definite mechanism of action of the compounds of this invention, but we think that the two quaternary salts of the 4-anilino-quinoline of 4-anilino-quinoline of the present invention and prior art are compared and suppress DNMT1 and/or suppress to have more specificity aspect the dna methylation in the cell, and this is because they have less charged substituting group or by group replacement season quinoline and/or pyridine functional groups with low pKa value. Therefore, the compounds of this invention should have the cytotoxicity of reduction, and the dna methylation of regulating more specifically in the cell is active.

1. the method for preparing the compounds of this invention

Compound of the present invention can prepare by the method shown in the following reaction scheme 1-6, and provides in specific embodiment.

In reaction scheme 1, wherein Y be CONH and Z be Q1 formula (I) compound preparation can by make 4-(pyridine radicals) pyridine and nitroaniline (2) reaction, with H2/Pd it is reduced to amine (3), then make itself and 4-nitrobenzoyl acid reaction, obtain acid amides (6), with its quaternized (for example, using MeOTs), obtain pyridine compounds (7). These compounds with iron powder/HCl reduction, are obtained amine (8), make itself and the coupling of 4-chloroquinoline and pass through in the case of necessary other processing, obtain the compound of formula (I).

(i)TsOH/180℃；(ii)H ₂/Pd/C/MeOH；(iii)5/SOCl ₂/ backflow, then 4/ pyridine/diox; (iv) DMF/MeOTs/20 ℃, then ion-exchange (v) Fe powder/EtOH/H₂O; (vi) 4-chloroquinoline/MeOH/cat.HCl/ refluxes

In reaction scheme 2; wherein Y is that NHCO and Z are that the compound of the formula (I) of Q1 can be prepared as follows; for example EDCI or CDI make N-acetyl-amino benzoic acid (9) and the reaction of 4-nitroaniline to use coupling agent; obtain acid amides (10); its usefulness is contained 1 of rare HCl; the hydrolysis of 4-diox obtains amine (11). Itself and 4-pyridine radicals pyridinium chloride are reacted, obtain amine (12). With its quaternized (for example, using MeOTs), obtain pyridine compounds (13) as mentioned above, it is reduced to amine (14), then as previously mentioned with the coupling of 4-chloroquinoline, and pass through in the case of necessary other processing, obtain the compound of formula (I).

Scheme 2

(i) EDCI/DMF/DMAP/4-nitroaniline; (ii) HCl/ diox/backflow (iii) p-TsOH/4-pyridine radicals pyridinium chloride/180 ℃

(iv) DMF/MeOTs; (i) Fe powder/EOH/H₂O/H ⁺/ then ion-exchange; (vi) 4-chloroquinoline/EtOH/H₂O/H ⁺/ reflux

In reaction scheme 3; wherein Y is that NHCO and Z are that the compound of the formula (I) of Q3 can be prepared as follows; make 4-chloroquinoline (15) and 4-acetyl-amino aniline (16) coupling; obtain anilino-quinoline (17); with its deblocking (deblocking), obtain anilino-quinoline (18). Make the reaction of acetylbenzoic acid (19) and aminoguanidine, obtain guanylhydrazones (guanylhydrazones) (20), use EDCI or other coupling agent to make itself and anilino-quinoline (18) coupling, obtain the compound of formula I.

(i) 2-aminoguanidine/MeOH/c.HC/ refluxes; (ii) EtOH/H₂O (2:1)/c.HC/ refluxes; (iii) 2N HC/EtOH/ refluxes; (iv) EDCI/DMAP/DMF/RT

In reaction scheme 4; wherein Y is that NHCO and Z are that the compound of the formula (I) of Q2 can be prepared as follows; make 4-anilino-quinoline (18) and N-acetylbenzoic acid (21) coupling; obtain acid amides (22); it is hydrolyzed to amine (23); then with 2-amino-6-chloro-4-methylpyrimidine reaction, obtain the compound of formula I.

(i) EDC1/DMAP/DMF/RT; (ii) Isosorbide-5-Nitrae-dioxs/HCl refluxes; (iii) 2-amino-6-chloro-4-methylpyrimidine/2-ethoxy ethanol/H⁺/ reflux

In reaction scheme, wherein Y is that CONH and Z are that the compound of the formula (I) of Q3 can be prepared as follows, and makes nitro-acetophenone (24) and aminoguanidine reaction, uses subsequently Fe/HCl (25) reduzate guanylhydrazones, obtains amine (26). Itself and 4-nitrobenzoyl chloride are reacted, and again product acid amides (27) is reduced to amine (28). Then make itself and the coupling of 4-chloroquinoline, obtain the compound of formula I.

Scheme 5

In reaction scheme 6, wherein Y is that CONH and Z are that the compound of the formula (I) of Q2 can be prepared as follows, make nitroaniline (2) and 2-amino-6-chloro-4-methylpyrimidine coupling, and with the product (27) that the Fe/HCl reduction obtains, obtain amine (28). Make itself and the coupling of 4-nitrobenzoyl chloride, and as previously mentioned with product acid amides (29) reduction, obtain amine (30). Make as mentioned above itself and the coupling of 4-chloroquinoline, obtain the compound of formula (I).

Quinoline of the present invention comprises the salt of the acceptable salt of its any pharmacy, ester or this ester or at any other compound that (directly or indirectly) its BA metabolin or residue can be provided when comprising people's animals administer. Therefore, for example, the disclosure also relates to pro-drug and the acceptable salt of pharmacy, the acceptable salt of the pharmacy of this pro-drug and other bioequivalence thing of the compounds of this invention.

Term " pro-drug " expression is prepared as the therapeutic agent of inactive form, and it in vivo or be converted into activity form (that is, medicine) in the cell under the effect of endogenous enzyme or other chemical reagent and/or condition. Especially; the prodrug form of quinoline of the present invention can prepare by forming one or more ester bonds with any oh group in the carboxylic organic compound of bag and the compound; perhaps according to WO 93/2451 or WO 94/26764 and United States Patent (USP) 5; disclosed method is prepared as SATE[(S-acetyl group-2-mercaptoethyl in 770,713) phosphate) derivative.

Term " the acceptable salt of pharmacy " refers to physiology and the acceptable salt of pharmacy of the compounds of this invention, that is, kept the biologically active of the hope of parent compound still not give the salt of undesirable toxicological action at this. Therefore, preferably get rid of the two quaternary salts of 4-anilino-quinoline.

The acceptable base addition salts of pharmacy forms with metal or amine, for example alkali and alkaline earth metal ions or organic amine. Example as cationic metal is sodium, potassium, magnesium, calcium etc. The example of the amine that is fit to is N, N '-dibenzyl-ethylenediamin, chloroprocanine, choline, diethanol amine, dicyclohexylamine, ethylenediamine, N-METHYL-ALPHA-L-GLUCOSAMINE and procaine are (referring to for example, the people such as Berge, ＂ Pharmaceutical Salts, ＂ J.of Pharma Sci., 1977,66,1-19). The base addition salts of described acid compound be by make free acid form and fully the required alkali of amount contact in a usual manner and form salt and prepare. Can by make salt contact a kind of acid and in a usual manner separated free acid make free acid form regeneration. Free acid form is slightly different from their salt forms separately aspect some physical property (for example, the solubility in polar solvent), but for purpose of the present invention, salt is identical with their free acids separately in other side.

As used in this article, " medicine addition salts " comprises the acceptable salt of pharmacy of a kind of sour form component of composition of the present invention. These comprise organic acid or the inorganic acid salt of amine. The acceptable salt of pharmacy that is fit to is as known in the art, comprises various inorganic acids and organic acid basic salt, such as for example, with inorganic acid for example hydrochloric acid, hydrobromic acid, sulfuric acid or phosphoric acid; With the amine sulfonic acid of organic carboxyl acid, sulfonic acid, thio-acid or phosphonic acids or N-replacement, for example acetic acid, propionic acid, glycolic acid, succinic acid, maleic acid, hydroxymaleic acid, citraconic acid, fumaric acid, malic acid, tartaric acid, lactic acid, oxalic acid, gluconic acid, glucosaccharic acid, glucuronic acid, citric acid, benzoic acid, cinnamic acid, mandelic acid, salicylic acid, 4-ASA, 2-phenoxy benzoic acid, Aspirin, pamoic acid, nicotinic acid or isonicotinic acid; With with amino acid, 20 kinds of a-amino acids that for example relate to during at synthetic protein at occurring in nature, for example glutamic acid or L-aminobutanedioic acid, and also has phenylacetic acid, methanesulfonic acid, ethyl sulfonic acid, 2-ethylenehydrinsulfonic acid, ethane-1,2-disulfonic acid, benzene sulfonic acid, 4-toluene sulfonic acide, naphthalene-2-sulfonic acid, naphthalene-1,5-disulfonic acid, 2-or 3-phoshoglyceric acid, Robison ester, N-cyclohexyl sulfamic acid (form of cyclamate), or other acid organic compound, for example ascorbic acid. The acceptable salt of the pharmacy of compound also can prepare with the acceptable cation of pharmacy. The acceptable cation of pharmacy that is fit to comprises alkali metal for well known to a person skilled in the art. Alkaline-earth metal, ammonium and quaternary ammonium cation. Carbonate or bicarbonate also are possible.

The present invention also comprises the compound through separating. Refer to that through the compound that separates compound accounts at least 10%, preferred 20%, more preferably 50% and most preferably 80% of the compound that is present in the mixture, and show when in biologic test, testing can detect (, statistically significant) direct or indirect dna methylation inhibition activity, the bisulfites restriction analysis that described biologic test for example makes up or COBRA (Xiong, Z.; Laird, P.W. Nucleic Acids Res.1997,25,2532-2534), radiolabeled methyl in conjunction with the test (Francis, K.T.; Thompson, R.W.; Krumdieck, C.L.Am. J.Clin.Nutr.1977,30,2028-2032) and in the people such as Ghoshal (2005) 11:4727-41 and the DNMT test partly described of following examples. Preferably, active with respect to the inhibition to DNMT3a and DNMT3b, compound selective of the present invention ground suppresses the activity of DNMT1.

2. Pharmaceutical preparation of the present invention

According to the present invention, compound of the present invention can be formulated as the acceptable composition of pharmacy, is used for the treatment of various diseases and the patient's condition.

The acceptable composition of pharmacy of the present invention comprise one or more compounds of the present invention and one or more nontoxic, pharmaceutically acceptable carrier and/or diluent and/or auxiliary agent and/or excipient (described carrier and/or diluent and/or auxiliary agent and/or excipient are referred to as " carrier " material in this article) and other active component as required.

Compound of the present invention is preferably the pharmaceutical compositions that is suitable for this approach by any administration, described method of administration such as following illustrational and depend on the patient's condition that will treat. Described compound and composition can be undertaken by multiple method of administration, for example: administration in administration in administration in the oral administration, parenterai administration, peritonaeum, intravenous administration, the artery, cutaneous penetration, sublingual administration, the muscle, rectally, transbuccally administration, intranasal administration, liposome administration, by inhalation, vagina administration, eye drops, by administration in local delivery administration (for example by conduit or support), subcutaneous administration, the fat, intra-articular administration or intrathecal drug delivery.

Described pharmaceutical preparation can randomly comprise the excipient with a certain amount of adding in addition, the amount of described excipient is enough to increase composition stable, keep product is solution form or the prevention side effect relevant with giving preparation of the present invention (for example, the stimulation of possible ulcer, blood vessel or exosmose). The example of excipient include but not limited to sweet mellow wine, sorbierite, lactose, glucose, cyclodextrin (for example, alpha-cyclodextrin, beta-schardinger dextrin-and gamma-cyclodextrin) and modification, unbodied cyclodextrin (for example hydroxypropyl-, hydroxyethyl-, glucityl-, malt-base-, maltotriose glycosyl (maltotriosyl-), carboxamide groups methyl-, carboxymethyl-, sulfo group butyl ether-and α, β of diethylamino-replacement-and gamma-cyclodextrin). Can use for this purpose cyclodextrin for example to derive from the Encapsin of Janssen Pharmaceuticals

Or equivalent.

For oral administration, described pharmaceutical composition can be for example form of tablet, capsule, suspension or liquid. Described pharmaceutical composition is preferably made the form of the dosage device that comprises the active component for the treatment of effective dose. The example of this dosage device is Tablet and Capsula. For therapeutic purposes, Tablet and Capsula can comprise the carrier of the routine except active component, and for example, adhesive is Arabic gum, gelatin, polyvinylpyrrolidone, sorbierite or bassora gum for example; Filler is calcium phosphate for example, glycine, lactose, cornstarch, sorbierite, or sucrose; Lubricant is dolomol for example, polyethylene glycol, silica or talcum; Disintegrant is farina for example, flavor enhancement or colouring agent, or acceptable wetting agent. The oral liquid prepared product that is generally solution, suspension, emulsion, syrup or the elixir form of moisture or oil-containing can comprise conventional additive, for example suspending agent, emulsifying agent, non-water reagent, anticorrisive agent, colouring agent and flavor enhancement. The example that is used for the additive of liquid prepared product comprises edible fat, lecithin, methylcellulose, methyl p-hydroxybenzoate or propylparaben, propane diols, sorbierite or the sorbic acid of Arabic gum, apricot kernel oil, ethanol, fractionated coconut oil, gelatin, glucose syrup, glycerine, hydrogenation.

For topical application, compound of the present invention can also be prepared as the suitable form be used to the mucous membrane that is applied to skin or nose and throat, and can be the form of creme, paste, liquid spray or inhalant, lozenge or throat paint. This topical formulations can comprise for example methyl-sulfoxide (DMSO) of compound in addition, to promote the surface infiltration of active component.

For the application of E ﹠ E, compound of the present invention can be used as liquid or the semi-liquid form prepared and exists in hydrophobicity or hydrophilic matrix, for example paste, creme, washing lotion, paint or pulvis.

For rectally, the form of the suppository that compound of the present invention can mix for the carrier (for example, cocoa butter, wax or other glyceride) with routine.

Perhaps, compound of the present invention can be for being used for when sending the powder type in suitable pharmaceutically acceptable carrier reconstruct.

Described pharmaceutical composition can pass through drug administration by injection. The preparation that is used for parenterai administration can be the isotonic sterile injection solution of moisture or non-water or the form of suspension. These solution or suspension can be from aseptic powder or the particle preparations that comprises one or more carriers, and described carrier is as described for the preparation that is used for oral administration. Compound can be dissolved in polyethylene glycol, propane diols, ethanol, corn oil, phenmethylol, sodium chloride and/or various buffer solution.

3. be used for giving the method for compound/composition of the present invention

Compound of the present invention or preparation can pass through any administration, are preferably the pharmaceutical compositions that is suitable for this approach, described method of administration such as following illustrational and depend on the patient's condition that will treat. Described compound or preparation can be by following method of administration administrations: administration in administration in administration in oral administration, parenterai administration, topical, the peritonaeum, intravenous administration, the artery, cutaneous penetration, sublingual administration, the muscle, rectally, transbuccally administration, intranasal administration, liposome administration, by inhalation, vagina administration, eye drops, by administration in local delivery administration (for example, by conduit or support), subcutaneous administration, the fat, intra-articular administration or intrathecal drug delivery. Compound of the present invention and/or composition can also be with the administration of agent for slow releasing type or co-administereds.

Compound of the present invention and/or composition can be with formulation administration and the co-administereds of any routine. Co-administered in the scope of the invention is defined as expression will be above the administration in parallel therapeutic process of a kind of therapeutic agent, in order to realize the clinical effectiveness of improvement. This co-administered can also carry out in time jointly, that is to say co-administered within the overlapping time period.

Compound of the present invention or comprise the composition of the compounds of this invention can be with 0.1-1000 mg/m²Dosage to host's administration, randomly be 1-200mg/m², randomly be 1-50mg/m², randomly be 1-40mg/m², randomly be 1-30mg/m², randomly be 1-20mg/m², or randomly be 5-30mg/m²。

Pharmaceutical preparation can and be selected from one or more member's co-administereds of following group with any conventionally form, and described group comprises infused fluid, therapeutic compound, trophism fluid, antimicrobial fluid, buffer solution and stabilizing agent.

The patient can accept described pharmaceutical preparation by intravenous. Preferred method of administration is intravenous infusion. Randomly, pharmaceutical preparation of the present invention can direct infusion, and need not prior reconstruct.

Because the preparation stability that increases, the patient can infusion medicament preparation 1,2,3,4,5 or more hours. The infusion time that prolongs can realize giving the flexibly timetable of therapeutic preparation.

As selecting or additionally, can regulating according to patient's needs speed and the volume of infusion. The infusion management of pharmaceutical preparation can be carried out according to existing flow process.

Pharmaceutical preparation can and be selected from following group the common infusion of one or more members with any conventionally form, and described group comprises infused fluid, therapeutic compound, trophism fluid, antimicrobial fluid, buffer solution and stabilizing agent. Randomly, can be with following therapeutic component and the common infusion of preparation of the present invention, described therapeutic component includes but not limited to medicine, biology medicament, interleukins, interferon, cell factor, immunomodulator and the monoclonal antibody of antineoplastic, alkylating agent, the medicine as the vitamin A acid superfamily member, antibiolics, hormone drug, plant origin.

Common infusion in the scope of the invention is defined as expression will be above a kind of therapeutic agent infusion in parallel therapeutic process, in order to realize the clinical effectiveness of improvement. This common infusion can be simultaneously, overlapping or sequentially carry out. In a specific example, the common infusion of pharmaceutical preparation and infused fluid can be undertaken by Y-connection.

The pharmacokinetics of the pharmaceutical preparation of intravenous administration is similar with metabolism to the pharmacokinetics of the compounds of this invention of intravenous administration with metabolism.

4. the therapeutic alliance of pharmaceutical composition of the present invention

Compound of the present invention or pharmaceutical preparation can be used for and the cytidine analog associating, and described cytidine analog is DAC (Decitabine) and 5-azepine-cytidine (azacitidine) for example.

Decitabine is the antagonist of its relevant natural nucleus glycoside (deoxycytidine). Unique architectural difference is that 5 in the cytimidine ring of Decitabine have nitrogen between these two kinds of compounds, and deoxycytidine is carbon in this position.

Decitabine has multiple pharmacological characteristic. At molecular level, it incorporates DNA into is the S stage dependent. At cellular level, Decitabine can Cell differentiation inducing activity and generation haematics toxicity.

The most important function of Decitabine is its ability that suppresses specifically and effectively dna methylation. As above described for cytosine methylation in the CpG island for example, cytosine methylation is the level that 5-methylcytosine occurs in DNA. In cell, Decitabine at first is converted into its activity form by deoxycytidine kinase, and the 5-azepine-deoxycytidine of phosphorylation, described deoxycytidine kinase are mainly synthetic in the S of cell cycle phase process. Decitabine is similar to natural substrate deoxycytidine to the affinity of the catalytic sites of deoxycytidine kinase. Be converted into its triguaiacyl phosphate form by deoxycytidine kinase after, Decitabine is incorporated among the DNA that is copying with the speed similar to natural substrate (dCTP). Bouchard and Momparler (1983) Mol.Pharmacol.24:109-114.

Decitabine is attached to has hypomethylation (hypomethylation effect) in the DNA chain. Each classification of the cell of differentiation has the different methylation patterns of oneself. After chromosome repeated, in order to preserve this methylation patterns, the 5-methylcytosine on the fundamental chain was used for directly methylating of complementary daughter DNA chain. 5 carbon of cytimidine are replaced with nitrogen hinder this normal dna methylation process. Replace 5-methylcytosine at methylated ad-hoc location with Decitabine and produce irreversible dnmt rna inactivation, the chances are owing to formed covalent bond between enzyme and Decitabine. The people such as Juttermann (1994) Proc.Natl.Acad.Sci.USA 91:11797-11801. By suppressing specifically dnmt rna (required enzyme methylates), the abnormal methylation that can prevent tumor suppressor gene.

Compound of the present invention or pharmaceutical preparation can be united use with histone deacetylase (HDAC); so that with transcribing of the further regulatory gene of cooperative mode; for example, be used for to recover transcribing by the gene of the supermethylation of histone and acetyl group silence.

Compound of the present invention or pharmaceutical preparation can be united use with histone deacetylase (HDAC); so that with transcribing of the further regulatory gene of cooperative mode; for example, be used for to recover transcribing by the gene of the supermethylation of histone and acetyl group silence.

HDAC plays important effect in the Transcriptional Silencing of gene. The amount of the acetyl group on the histone is controlled by the opposite activity of two types of enzymes, and described two types enzyme is histone acetyl based transferase (HAT) and histone deacetylase (HDAC). The substrate of these enzymes comprises the e-amino group of the lysine residue of the amino terminal afterbody that is arranged in histone H 3, H4, H2A and H2B. These amino acid residues are by the HAT acetyl group, and by the HDAC deacetylation. When removing Acetyl Groups by HDAC from istone lysine, lysine residue recovers positive charge, thus condensing nucleosomal structure and make the gene silencing that wherein comprises. Therefore, in order to activate these genes by the deacetylase silence of histone, should suppress the activity of HDAC. By suppressing HDAC, histone by acetyl group and the DNA that is wrapped in tightly around the histone core of deacetylation lax. Dna structure open the expression that causes specific gene.

Except the deacetylation of histone, HDAC can also express by deacetylation transcription factor regulatory gene, for example, and p53 (tumor suppressor gene), GATA-1, TFIIE and TFIIF. Gu and Roeder (1997) Cell 90:595-606 (p53); With people (1998) Nature 396:594-598 (GATA-1) such as Boyes. HDAC also participates in Cycle Regulation by for example transcription repression, and described transcription repression is regulated by the RB tumor inhibitor protein of mobilizing HDAC. The people such as Brehm (1998) Nature 391:597-601. Therefore, the inhibition of HDAC should activate for example expression of p53 and RB of tumor suppressor gene, thereby and promotes by these gene induced cell growth inhibitions, differentiation and apoptosis.

As mentioned above, the unusual Transcriptional Silencing of many genes (for example, tumor suppressor gene) is directly relevant with the pathogenesis of cancer and Other diseases. The cytimidine residue methylates and removes two main mechanism that Acetyl Groups is gene silencing from histone among the DNA. Because methylating and/or histone deacetylase of cancer related gene, the expression of these genes are suppressed or are complete reticent. Simultaneously, the expression of these genes is that growth inhibition, differentiation and/or the apoptotic cell death of Induction Transformation cell is needed. The non-activity of these genes in transformant causes the uncontrolled propagation of these cells, and it causes cancer at last.

By with compound/composition of the present invention and hdac inhibitor combination, the needed gene of Induction Transformation cell growth inhibition, differentiation and cell death is brought back to life. Compound/composition of the present invention suppresses the dna methylation of described gene, and the dna methylation in regulatory region particularly causes the transcription activating of described gene thus. Simultaneously, hdac inhibitor suppresses the deacetylase of the histone in the nucleosome core of described gene, causes thus the net increase of the deacetylation of histone, and it activates again transcribing of described gene. By utilizing the mechanism of these two kinds of complementations, described therapeutic alliance can more effectively and ideally recover genetic transcription in collaborative mode. Every kind of inhibitor of less amount gave the relevant possible side effect of high dose inhibitor and improves therapeutic index with system thereby reduce when the therapeutic alliance with cooperative effect should need than independent the use.

Many anticarcinogens are brought into play its antitumaous effect by the signal transduction cascade that triggering relates to the protein of being encoded by these tumor suppressor genes. Owing to the insufficient expression of these genes in cancer cell, can reduce tempestuously or fully eliminate the antitumaous effect of these antineoplastics. By by the reactivation of these genes of the outer genetic silencing of dna methylation and histone deacetylase or express again; the intrinsic defense mechanism of health is mobilized, by recovering the tumor suppression function of cancer cell is resisted disease in response to the signal that is sent by the anticarcinogen that gives. This stimulation of the tumor suppression function that health is intrinsic should cause that the dosage demand to anticarcinogen reduces, thereby produces higher drug therapy index (that is, larger effectiveness and lower toxicity).

The inhibitor of HDAC comprises but is not limited to following structured sort: 1) hydroxamic acid, 2) cyclic peptide, 3) benzamides and 4) short-chain fat acids.

The example of hydroxamic acid and hydroxamic acid derivs includes but not limited to TSA (TSA), cork anilid hydroxamic acid (SAHA), oxamflatin, the two hydroxamic acid (SBHA) of suberic acid, the two hydroxamic acid (CBHA) of an o-carboxy cinnamic acid and pyroxamide. TSA separates and to obtain people (1976) J. Antibiot (Tokyo) 29:1-6 such as () Tsuji as antifungal antibiotic and finds is effective inhibitor people (1990) J.Biol.Chem.265:17174-17179 such as () Yoshida of mammal HDAC. The discovery that TSA tolerance clone has the HDAC of change has proved that this kind of enzyme is the important target of TSA. Other hdac inhibitor based on hydroxamic acid, SAHA, SBHA and CBHA are the synthetic compounds that can suppress HDAC in external or body with micro-molar concentration or lower concentration. The people such as Glick (1999) Cancer Res.59:4392-4399. These all have the architectural feature an of necessity based on the hdac inhibitor of hydroxamic acid: by hydrophobicity methylene interval base (for example, 6 carbon length) it is terminal to be connected in the hydroximic acid of polarity of another polarity position, described another polarity position is connected in terminal hydrophobic group (for example, phenyl ring). The compound with this essential characteristic of exploitation also falls in the scope of the hydroxamic acid that can be used as hdac inhibitor.

Cyclic peptide as hdac inhibitor mainly is the tetrapeptide of ring-type. The example of cyclic peptide includes but not limited to trapoxin A, apicidin and FR901228. Trapoxin A comprises a 2-amino-8-oxo-9, the tetrapeptide of the ring-type of 10-epoxy-capryl (AOE) part. The people such as Kijima (1993) J.Biol.Chem.268:22429-22435. Apicidin is a kind of fungal metabolite, and it is active and active at nanomole level control of the concentration HDAC that it shows antiprotozoals strong, wide spectrum. The people such as Darkin-Rattray (1996) Proc.Natl. Acad.Sci.USA.93; 13143-13147. FR901228 is a kind of depsipeptide that separates from Chromobacterium violaceum (Chromobacterium violaceum), and has shown at micromolar concentrations inhibition HDAC active.

The example of benzamides includes but not limited to MS-27-275. The people such as Saito (1990) Proc.Natl.Acad.Sci.USA.96:4592-4597. The example of short-chain fat acids includes but not limited to butyric acid class (for example, butyric acid, arginine butyrate and phenyl butyrate (PB)). The people such as Newmark (1994) Cancer Lett.78:1-5; With people (1997) Anticancer Res.17:3972-3973 such as Carducci. In addition, having shown depudecin people (1998) Proc.Natl.Acad. Sci.USA.95:3356-3361 such as () Kwon that suppresses HDAC in micromolar concentrations also falls in the scope of histone deacetylase inhibitors of the present invention.

Compound of the present invention or pharmaceutical preparation can also be used for and other therapeutic component associating, and described other therapeutic component includes but not limited to medicine, biology medicine, interleukins, interferon, cell factor, immunomodulator and the monoclonal antibody of antineoplastic, alkylating agent, the medicine as the retinoid superfamily member, antibiolics, hormone drug, plant origin.

In one embodiment, alkylating agent is used for be used in combination with compound/preparation of the present invention and/or adds alkylating agent for compound of the present invention/preparation. The example of alkylating agent includes but not limited to two chloroethyl amine (nitrogen mustardses, for example, Chlorambucil, endoxan, ifosfamide, chlormethine, melphalan, uracil mustard), aziridines (for example, phosphinothioylidynetrisaziridine), the alkyl ketone sulfonic acid esters (for example, busulfan), nitrosoureas (for example, BCNU, lomustine, streptozotocin), non-classical alkylating agent (hexamethyl melamine, Dacarbazine and procarbazine), platinum compounds (carboplatin and cis-platinum).

In another embodiment, cis-platinum, carboplatin or endoxan are used for be used in combination with compound/preparation of the present invention and/or add cis-platinum, carboplatin or endoxan for compound of the present invention/preparation.

In another embodiment, the member of retinoid superfamily is used for being used in combination and/or adding for compound of the present invention/preparation with compound/preparation of the present invention the member of retinoid superfamily. Retinoid be structurally with function on relevant molecule family, it derives from vitamin A (alltrans retinol) or relevant with it. The example of retinoid includes but not limited to alltrans retinol, all-trans retinoic acid (vitamin A acid), Accutane (Accutane) and 9CRA.

In another embodiment, hormone drug is used for being used in combination with compound/preparation of the present invention and/or adds hormone drug for compound of the present invention/preparation. The example of this hormone drug for synthetic estrogen (for example, diethylstilbestrol), antiestrogen (for example, TAM, Toremifene, fluoxymesterone and Raloxifene), antiandrogen (Bicalutamide, Nilutamide, Flutamide), aromatase inhibitor (for example, aminoglutethimide, Anastrozole and tetrazolium), ketoconazole, goserelin acetate, leuprorelin acetate, megestrol acetate and mifepristone.

In another embodiment, the medicine of plant-origin is used for being used in combination and/or adding for compound of the present invention/preparation with compound/preparation of the present invention the medicine of plant-origin.The example of the medicine of plant origin includes but not limited to that vinca alkaloids (for example, vincristine(VCR), vinealeucoblastine(VLB), vindesine, vinzolidine and vinorelbine), camptothecine (20 (S)-camptothecine, 9-nitro-20 (S)-camptothecine and 9-amino-20 (S)-camptothecine), podophyllotoxin (for example, Etoposide (VP-16) and teniposide (VM-26)) and taxanes (for example, taxol and docetaxel).

In another embodiment, the biology medicament is used for adding the biology medicament with the combination of compound of the present invention/preparation and/or for compound of the present invention/preparation, described biology medicament is immune modulator for example, for example cytokine, the monoclonal antibody to tumor-resistant antigen, tumor suppressor gene and cancer vaccine.

The example that can be used for being used in combination with compound/preparation of the present invention and/or add the interleukin-of compound/preparation of the present invention to includes but not limited to interleukin II (IL-2) and interleukin-4 (IL-4), interleukin 12 (IL-12).The example that can be used for the Interferon, rabbit that is used in combination with Decitabine-glycerin preparation includes but not limited to interferon alpha, interferon beta (fibroblast Interferon, rabbit) and interferon-gamma (fibroblast Interferon, rabbit).The example of this cytokine includes but not limited to erythropoietin (epoietin), granulocyte-CSF (filgrastim) and granulocyte, scavenger cell-CSF (Sargramostim).The immunomodulator that is different from cytokine includes but not limited to bacille Calmette-Guerin vaccine (bacillusCalmette-Guerin), LEVAMISOLE HCL and Sostatin.

The example to the monoclonal antibody of tumor-resistant antigen that can be used for being used in combination with preparation of the present invention includes but not limited to

(Trastruzumab), (sharp appropriate Xidan is anti-),

(anti--CD33 antibody) and

(anti-CD 52 antibody).

In another embodiment, kinase inhibitor is used for making up with compound of the present invention/preparation and/or adding compound formulation of the present invention to, be used for the treatment of and the abnormal kinase diseases associated.

In a variant, described tyrosine kinase inhibitor be imatinib mesylate (for example,

Imatinib mesylate is a kind of protein tyrosine kinase inhibitors, and it is suppressed among the CML by the unusual Bcr-Ab1 Tyrosylprotein kinase that produces of Philadelphia chromosome.Imatinib mesylate realizes this inhibition result by the Adenosine Triphosphate combining site that is incorporated into the Bcr-Ab1 Tyrosylprotein kinase, the phosphorylation and relevant vicious transformation of prevention substrate.By suppressing this kinases, think that imatinib mesylate suppresses cell proliferation and apoptosis-induced.People such as T.Schindler (2000) Science 289:1938-1942.

In another variant, described kinases is a kind of serine-threonine kinase, for example the Raf kinases; And described kinase inhibitor is BAY 43-9006.

In another variant, kinases is for example Raf-mitogen-activated protein kinase (MEK) and protein kinase B (Akt) kinases of protein kinase.

In another variant, described kinases is extracellular signal-regulated kinase (ERK).The example of ERK inhibitor includes but not limited to PD98059, PD184352 and U0126.

In another variant, described kinases be phosphatidylinositols 3 '-kinases (PI3K).The example of the inhibitor of PI3K includes but not limited to LY294002.

In specific variant, described kinases is a Tyrosylprotein kinase.Described Tyrosylprotein kinase can be receptor tyrosine kinase and nonreceptor tyrosine kinase.

The example of receptor tyrosine kinase includes but not limited to Epidermal Growth Factor Receptor Family (EGFR), platelet derived growth factor receptor (PDGFR) family, vascular endothelial growth factor receptor (VEGFR) family, trk C (NGFR) family, fibroblast growth factor acceptor family (FGFR), Insulin Receptor Family, ephrin receptor family, Met family and Ror family.

The example of Epidermal Growth Factor Receptor Family includes but not limited to HER1, HER2/Neu, HER3 and HER4.

The example of the inhibitor of Epidermal Growth Factor Receptor Family includes but not limited to

ZD1839

PD168393, CI1033, IMC-C225, EKB-569 and be covalently bonded in the inhibitor of the Cys residue of receptor tyrosine kinase.

Include but not limited to cancer, lung cancer and the nonsmall-cell lung cancer of epithelial knurl, malignant tumour, the upper respiratory tract-digestive tube (upperaerodigestive tract) with the example of the active unusual diseases associated of Epidermal Growth Factor Receptor Family.

The example of vascular endothelial growth factor receptor family includes but not limited to VEGFR1, VEGFR2 and VEGFR3.

The example of the inhibitor of vascular endothelial growth factor receptor family includes but not limited to SU6668.

Include but not limited to solid tumor and the tumour that metastasis tendency is arranged with the active example of diseases associated unusually of vascular endothelial growth factor receptor family.

The example of trk C family includes but not limited to trk, trkB and trkC.

The example of the inhibitor of trk C family includes but not limited to CEP-701, CEP-751 and indocarbazole compound.

Include but not limited to prostate cancer, colorectal carcinoma, papillary carcinoma and thyroid carcinoma, neuroma and osteoblastoma with the active example of diseases associated unusually of trk C family.

The example of Met family includes but not limited to Met, TPR-Met, Ron, c-Sea and v-Sea.

Include but not limited to the squamous cell carcinoma of the ingrown tumour of infectivity, malignant tumour, thyroid papillary carcinoma, colorectal carcinoma, kidney, carcinoma of the pancreas, ovarian cancer, head and neck with example from the active diseases associated of the receptor tyrosine kinase of Met family.

The example of nonreceptor tyrosine kinase includes but not limited to c-kit family, Src family, Fes family, JAK family, Fak family, Btk family, Syk/ZAP-70 family and Ab1 family.

Example from the nonreceptor tyrosine kinase of Src family includes but not limited to Src, c-Src, v-Src, Yes, c-Yes, v-Yes, Fyn, Lyn, Lck, Blk, Hck, Fgr, c-Fgr, v-Fgr, p561ck, Tk1, Csk and Ctk.

Example from the inhibitor of the nonreceptor tyrosine kinase of Src family includes but not limited to SU101 and CGP57418B.

Include but not limited to mammary cancer, malignant tumour, myelomatosis, leukemia and neuroblastoma with example from the active diseases associated of the nonreceptor tyrosine kinase of Src family.

Example from the nonreceptor tyrosine kinase of Fes family includes but not limited to c-fes/fps, v-fps/fes, p94-c-fes-related protein and Fer.

And include but not limited to the tumour in a tumour in matter source and green blood source from the example of the active diseases associated of the nonreceptor tyrosine kinase of Fes family.

Example from the nonreceptor tyrosine kinase of JAK family includes but not limited to Jakl, Jak2, Tyk2 and Jak3.

Include but not limited to member, synthetic component AG490, dimethoxyquinazoline compound, the 4-(phenyl)-amino-6 of tyrphostin (tyrphostin), CIS/SOCS/Jab family from the example of the inhibitor of the nonreceptor tyrosine kinase of JAK family, 7-dimethoxyquinazoline, 4-(4 '-hydroxy phenyl)-amino-6,7-dimethoxyquinazoline, 4-(3 '-bromo-4 '-hydroxy phenyl)-amino-6,7-dimethoxyquinazoline and 4-(3 ', 5 '-two bromo-4 '-hydroxy phenyl)-amino-6, the 7-dimethoxyquinazoline.

And include but not limited to the tumour in a tumour in matter source and green blood source from the example of the active diseases associated of the nonreceptor tyrosine kinase of JAK family.

Example from the nonreceptor tyrosine kinase of Fak family includes but not limited to Fak and CAK β/Pyk2/RAFTK.

Example from the inhibitor of the nonreceptor tyrosine kinase of Fak family includes but not limited to dominant negative mutant S1034-FRNK; Metabolite FTY720 and FAK antisense oligonucleotide ISIS 15421 from Isaria sinclarii.

Include but not limited to the tumour in human malignant tumour, the tumour that metastasis tendency is arranged and green blood source with example from the active unusual diseases associated of the nonreceptor tyrosine kinase of Fak family.

Example from the nonreceptor tyrosine kinase of Btk family includes but not limited to Btk/Atk, Itk/Emt/Tsk, Bmx/Etk and Itk, Tec, Bmx and Rlk.

Example from the inhibitor of the nonreceptor tyrosine kinase of Btk family includes but not limited to alpha-cyano-beta-hydroxy-Beta-methyl-N-(2, the 5-dibromo phenyl) acrylamide.

Include but not limited to B pedigree leukemia and lymphoma with example from the active unusual diseases associated of the nonreceptor tyrosine kinase of Btk family.

Example from the nonreceptor tyrosine kinase of Syk/ZAP-70 family includes but not limited to Syk and ZAP-70.

Example from the inhibitor of the nonreceptor tyrosine kinase of Syk/ZAP-70 family includes but not limited to piceatannol, 3,4-dimethyl-10-(3-aminopropyl)-9-dihydroketoacridine oxalate, dihydroketoacridine related compound, Lys-Leu-Ile-Leu-Phe-Leu-Leu-Leu[SEQ ID NO:1) peptide and comprise the peptide of Lys-Leu-Ile-Leu-Phe-Leu-Leu-Leu motif.

And from the nonreceptor tyrosine kinase of Syk/ZAP-70 family the example of active unusual diseases associated include but not limited to the tumour in benign mammary cancer, mammary cancer and a matter source.

5. The indication of compound of the present invention or pharmaceutical composition

Pharmaceutical preparation of the present invention can be used for the treatment of various diseases, is preferred for treating those diseases relevant with unusual dna methylation.

Can use the preferred indication of pharmaceutical preparation of the present invention treatment to comprise often to relate to and not expect or those of uncontrolled cell proliferation.This indication comprises innocent tumour; Various types of cancers, for example primary tumo(u)r and metastases; Restenosis (for example, coronary artery, carotid artery and cerebral lesion); The hematology illness; The abnormal stimulation of endotheliocyte (atherosclerosis); The infringement that health is caused owing to perform the operation; Unusual wound healing; Unusual vasculogenesis; Produce the disease of tissue fibrosis; The repeatable motion illness; The illness of the tissue of non-height vascularization; With the proliferative reaction relevant with organ transplantation.

Usually, the cell in the innocent tumour keeps its differentiating characteristic and is not to divide in complete uncontrolled mode.Innocent tumour normally localize with non-metastatic.The carcinoid particular type that can use the present invention to treat comprises vascular tumor, hepatic cell adenoma, porousness vascular tumor, focal nodular hyperplasia disease, acoustic tumor, neurofibroma, cholangioadenoma, bile duct cystanoma, fibroma, lipoma, leiomyoma, mesothelioma, teratoma, myxoma, trifle reproducibility hyperplasia, granular conjunctivitis and botryomycosis hominis.

Malignant cell becomes undifferentiated, to the not response of the growth control signal of health, and breeds in uncontrolled mode.Malignant tumour is invasive and can be diffused into position far away (transfer).Malignant tumour is divided into two classes usually: former with secondary.Primary tumo(u)r directly appears in the tissue of wherein finding them.Secondary tumor or metastatic tumor are other the local tumours that still is diffused into organ far away now that derives from health.The common approach that shifts be direct growth in adjacent structure, by blood vessel or the lymphsystem diffusion, and track is along the gap (peritoneal fluid, cerebrospinal fluid etc.) of tissue plane and health.

Can use that the present invention treats former or the cancer of secondary or the particular type of malignant tumour comprise mammary cancer, skin carcinoma, osteocarcinoma, prostate cancer, liver cancer, lung cancer, nonsmall-cell lung cancer, the cancer of the brain, laryngocarcinoma, carcinoma of gallbladder, carcinoma of the pancreas, the rectum cancer, parathyroid carcinoma, thyroid carcinoma, adrenal carcinoma, nervous center is organized cancer, head and neck cancer, colorectal carcinoma, cancer of the stomach, bronchogenic carcinoma, and kidney, rodent cancer, ulcer type and nipple type squamous cell carcinoma, the transitivity skin carcinoma, osteosarcoma, Ewing sarcoma, non-Hodgkin lymphomas (reticulum cell sarcoma), myelomatosis, giant cell tumor, small cell lung tumor, gallbladdergallstonecholetithiasis, islet cell tumor, primary brain tumors, acute and chronic lymphocytic knurl and granulocyte knurl, the hair cell knurl, adenoma, hyperplasia, medullary carcinoma, pheochromocytoma, mucous membrane neuroma (mucosal neuronms), enteron aisle gangliocytoma (intestinal ganglloneuromas), hyperplasia corneal nerve knurl, marfanoid habitus knurl, Wilm ' s knurl, spermocytoma, ovarian tumor, leiomyoma, the cervical atypism hyperplasia, and carcinoma in situ, neuroblastoma, retinoblastoma, soft tissue sarcoma, the carcinoid malignant knurl, partial skin lesion, cutaneous T cell lymphoma (mycosisfungoides), rhabdosarcoma, Kaposi, osteogenic sarcoma and other sarcoma, pernicious hypercalcemia, the nephrocyte tumour, polycythemia vera (polycythermiavera), gland cancer, glioblastoma multiforme (glioblastoma multiforma), leukemia, lymphoma, malignant melanoma, epidermoid carcinoma, with other cancer and sarcoma.

The hematology illness comprises can cause for example various leukemic hemocyte misgrowths of hemocyte dysplasia variation and haematological malignancies.The example of hematology illness includes but not limited to acute myelogenous leukemia, acute promyelocytic leukemia, acute lymphoblastic leukemia, chronic graininess leukemia, spinal cord heteroplasia syndrome and sickle cell disease.

Acute myelogenous leukemia (AML) is a modal acute leukemia type in the grownup.The genetic disorder of several successions raises relevant with immune deficiency state with the risk of AML.These comprise the illness with the dna stability defective, described dna stability defective causes rhexis at random, for example the agamaglobulinemia that is connected with X-of Bloom syndrome, Fanconi anemia, Li-Fraumeni kindreds, ataxia-telangiectasia.

Acute promyelocytic leukemia (APML) has been represented the different subgroups of AML.This hypotype is characterised in that and comprises 15; The promyelocyte parent cell of 17 chromosome translocations.This transposition causes the generation of being transcribed by the fusion that retinoic acid receptor (RAR) and sequence PML form.

Acute lymphoblastic leukemia (ALL) is the heterology disease (heterogenerous disease) with the different Clinical symptoms that shown by various hypotypes.Proved in ALL that the cytogenetics that takes place once more is unusual.Modal cytogenetics is 9 unusually; 22 transpositions.The Philadelphia chromosome that produces has been represented the prognosis of patient's difference.

Chronic graininess leukemia (CML) is the clone myeloproliferative disease of pluripotent stem cell.CML is characterised in that the specific chromosome abnormalty that relates to karyomit(e) 9 and 22 transpositions, produces Philadelphia chromosome.Ionizing rays is relevant with the formation of CML.

Spinal cord heteroplasia syndrome (MDS) is owing to one or more hemopoietic cells that the dysplasia that comprises myeloid cell series, erythron and megakaryocytic series changes are one group of heterology clone hemopoietic stem cell illness that existence that sour dysplasia changes causes.These variations cause one or more the cytopenia in described three pedigrees.The patient who suffers from MDS is generation and anemia, neutropenia (infection) or thrombopenia (hemorrhage) complications associated with arterial system typically.Usually, have an appointment 10% among the MDS patient to about 70% generation acute leukemia.

Because the abnormal cell proliferation that systemic infringement is caused at intra-operative may be because various surgical procedures comprise that operation on joint, gut surgery and cheloid scab.The disease that produces the fibrosis tissue comprises pulmonary emphysema.The repeatable motion illness that can use the present invention to treat comprises carpal tunnel syndrome.Can use the example of the cell proliferative disorders that the present invention treats to be bone tumor.

The proliferative reaction relevant with organ transplantation that can use the present invention to treat comprises the proliferative reaction that helps possible organ rejection or related complication.Particularly, this proliferative reaction can occur in the migration process of heart, lung, liver, kidney and other body member or tract.

The unusual vasculogenesis that can use the present invention to treat comprises those unusual vasculogenesis of following following disease: rheumatoid arthritis, cerebral edema that ischemic damage and reperfusion is relevant and damage, the cortex ischemic, ovary hyperplasia and blood vessel excessively form (hypervascularity), (polycystic ovary syndrome), endometriosis, psoriasis, diabetic retinopathy (diabetic retinopaphy), with other the eye vasculogenesis disease (for example, the retinopathy of premature labor (retrolental fiber formation)), myodegeneration, corneal graft rejection, neuroscular glaucoma and Oster Webber syndromes.

Growth with unusual diseases associated needs of vasculogenesis or induction of vascular.For example, the cornea vasculogenesis comprises three phases: the ripe and degeneration of latent period, active neovascularization and blood vessel before the blood vessel.The characteristic of the various angiogenic factors and mechanism await to disclose, and comprise struvite factor of replying, for example white corpuscle, thrombocyte, cytokine and eicosanoid, perhaps unidentified plasma component.

In another embodiment, pharmaceutical preparation of the present invention can be used for the treatment of and not expect or the disease of unusual associated angiogenesis.Described method comprise to suffer from do not expect or the patient of vasculogenesis give independent or with the pharmaceutical preparation of the present invention of antitumour drug combination, wherein said antitumour drug is subjected to the disadvantageous effect of high-level dna methylation as the activity in vivo of antitumour drug.The concrete dosage that suppresses these required medicines of vasculogenesis and/or angiogenic disease can depend on the severity of the patient's condition, route of administration and can be by the correlative factor of attending doctor's decision.Usually, acceptance and effectively per daily dose be the amount that is enough to effectively suppress vasculogenesis and/or angiogenic disease.

According to this embodiment, pharmaceutical preparation of the present invention can be used for the treatment of the various diseases with the associated angiogenesis of not expecting, for example neovascularization of retina/choroidal neovascularization and cornea.The example of retina/choroidal neovascularization includes but not limited to the Bests disease, myopia, look nest, the Stargarts disease, osteitis deformans, venous occlusion, arterial occlusion, sickle cell disease, sarcoid, syphilis, pseudoxanthoma elasticum carotid artery obstruction disease (abostructive disease), chronic uveitis/hyalitis, mycobacterial infections, Lyme disease, systemic lupus erythematous, the retinopathy of premature labor, eales disease, diabetic retinopathy, macular degeneration, the Bechets disease, cause the retinitis or uvaeformis infection, the ocular histoplasmosis of supposing, pars planitis, chronic retinal detachment, hyperviscosity syndrome, toxoplasmosis, wound and laser infectious-related complication, with flush (rubesis) diseases associated (neovascularization of angle (angle)) and the disease (form of ownership that comprises proliferative vitreoretinopathy) that causes by the abnormality proliferation of vascular tissue or fibrous tissue.The example of cornea neovascularization includes but not limited to epidemic keratoconjunctivitis, vitamin(e) A deficiency, contact lens excessively use, atopy keratitis, the high-grade marginal keratitis, pteryium keratitis sicca, sjogrens, acne erythematosa, phylectenulosis, diabetic retinopathy, the retinopathy of premature labor, corneal graft rejection, mooren's ulcer, the Terrien edge degradation, the marginality keratolysis, polyarteritis, the Wegener sarcoidosis, scleritis, the periphigoid radial keratotomy, neovascular glaucoma and retrolental fibroplasia, syphilis, mycobacterial infections, lipid is degenerated, chemical burns, bacterium ulcer, fungi ulcer, herpes-ness progenitalis infection, zoster infects, protozoal infections and Kaposi sarcoma.

In another embodiment, pharmaceutical preparation of the present invention can be used for the treatment of and the unusual relevant chronic inflammation disease of vasculogenesis.Described method comprise to suffer from give independent with the patient of the unusual relevant chronic inflammatory disease of vasculogenesis or with the pharmaceutical preparation of the present invention of antitumour drug combination, wherein said antitumour drug is subjected to the disadvantageous effect of high-level dna methylation as the activity in vivo of antitumour drug.The continuous formation that described chronic inflammatory diseases depends on the capillary vessel rudiment keeps the interior stream of inflammatory cell.The interior stream of inflammatory cell and existence produce granuloma, and keep the chronic inflammatory diseases sexual state thus.Use pharmaceutical preparation of the present invention to suppress vasculogenesis and can prevent the formation of granuloma, thereby alleviate this disease.The example of chronic inflammatory disease includes but not limited to inflammatory bowel for example Crohn's disease and ulcerative colitis, psoriasis, sarcoidosis and rheumatoid arthritis.

Inflammatory bowel for example Crohn's disease and ulcerative colitis is characterised in that chronic inflammatory diseases and vasculogenesis in GI a plurality of positions.For example, Crohn's disease exists as chronic saturating wall inflammatory diseases, and its most common terminal ileum and colon of influencing still also may be present in GI any part and perianal region from the mouth to the anus.Cd patient has the chronic diarrhoea relevant with stomachache, fever, apocleisis, body weight loss and abdominal distension usually.Ulcerative colitis also is a kind of chronic, nonspecific, struvite and ulcerative disease, and it appears in the mucous membrane of colon and is characterised in that the diarrhoea that has blood.Normally GI chronic granulomatous inflammation causes these inflammatory bowels by spreading all over, and comprises the new capillary vessel rudiment that cylinder centered on by inflammatory cell.Suppressing vasculogenesis by pharmaceutical preparation of the present invention should suppress forming of rudiment and prevent forming of granuloma.Inflammatory bowel also shows the outer performance of intestines, for example skin lesion.This infringement is characterised in that inflammation and vasculogenesis, and can take place the many positions beyond gi tract.Reducing vasculogenesis by pharmaceutical preparation of the present invention should reduce the interior stream of inflammatory cell and prevent infringement to form.

Sarcoid being characterised in that as another kind of chronic inflammatory disease is a kind of multisystem granuloma illness.The granuloma of this disease can be formed in the health Anywhere, and thus, symptom depends on whether the position of granuloma and disease are active.Granuloma is produced by the angiogenic capillary vessel rudiment of the long run supply that inflammatory cell is provided.The pharmaceutical preparation of the application of the invention suppresses vasculogenesis, can suppress this granuloma and form.Psoriasis also is a kind of chronic and inflammatory diseases recurrent, it is characterized in that the papule and the plaque of all size.The alleviation of using pharmaceutical preparation of the present invention to treat to prevent to keep the formation of the required neovascularity of characteristic pathology and symptom is provided as the patient.

Rheumatoid arthritis (RA) also is a kind of chronic inflammatory diseases, it is characterized in that nonspecific inflammation in periphery joint.Think that vasculogenesis has taken place the blood vessel in the knuckle synovia internal layer.Except forming new blood vessel network, endotheliocyte discharges the factor and the reactive oxygen species that causes pannus growth and cartilage destruction.The factor that relates to vasculogenesis can promote and help to keep the chronic inflammatory state of rheumatoid arthritis on one's own initiative.Use independent or treat the alleviation that can prevent to keep the formation of the required neovascularity of chronic inflammatory diseases and symptom is provided as RA patient with the pharmaceutical preparation of the present invention of other anti-RA medicine associating.

In another embodiment, pharmaceutical preparation of the present invention can be used for the treatment of and oxyphorase resulting anomaly diseases associated.Described method comprises that the patient to suffering from oxyphorase resulting anomaly diseases associated gives pharmaceutical preparation of the present invention.Because the mechanism that is incorporated among the DNA relates to the DNA hypomethylation, the preparation that comprises Decitabine stimulates foetal haemoglobin synthetic.Include but not limited to sickle cell disease and β-thalassemia with the example of oxyphorase resulting anomaly diseases associated.

In another embodiment, pharmaceutical preparation of the present invention can be used to control intracellular genetic expression.Described method comprises that the patient to suffering from the unusual diseases associated of gene expression dose gives pharmaceutical preparation of the present invention.Dna methylation is relevant with the control of genetic expression.Particularly, promotor methylate or promotor near methylate and suppress to transcribe, and demethylation recovers to express.The example of the possible application of described mechanism includes but not limited to growth-inhibiting, apoptosis induced and the cytodifferentiation of therapeutic regulation.

Can be used for the treatment of the differentiation of purpose inducing cell by the promoted gene activation of pharmaceutical preparation of the present invention.By hypomethylated machine-processed inducing cell differentiation.The example of the differentiation of form and function include but not limited to be divided into form myocyte, sarcotubules, the cell of red corpuscle and lymph pedigree.

Although described and narrated exemplary of the present invention, but it will be apparent to one skilled in the art that, can carry out multiple change, improvement or transformation to described the present invention herein, but not break away from spirit of the present invention.Therefore, all this changes, improvement and transformation all are considered within the scope of the present invention.

Embodiment

Following examples are representatives of the present invention, and the detailed method that is used to prepare The compounds of this invention is provided.In these embodiments, ultimate analysis is at University of Otago, Dunedin, and the Microchemical Laboratory of NZ carries out.On the Electrothermal2300 melting point apparatus, measure fusing point.On Bruker Avance-400 spectrograph, obtain with 400MHz ¹H NMR spectrum and obtain 13C NMR spectrum with 100MHz, reference data is Me4Si.Measure mass spectrum on the VG-70SE mass spectrograph, use the ionization potential of 70eV, nominal resolving power is 1000.Obtain high-resolution spectroscopy in 3000,5000 or 10000 nominal resolving power as required.Except as otherwise noted, all spectrum all are to use PFK to obtain as benchmark by electron-bombardment (EI).Except as otherwise noted, on silica gel (Merck 230-400 order), carry out column chromatography.

Embodiment A.

The preparation of 1-methyl-4-(3-{[4-(4-quinolyl amino) benzoyl] amino } anilino) pyridinium chloride (compd A)

(i) TsOH/180 ℃; (ii) H ₂/ Pd/C/MeOH; (iii) 4-nitrobenzoyl chloride/pyridine/diox; (iv) DMF/MeOTs/20 ℃, ion-exchange then; (v) Fe powder/EtOH/H ₂O; (vi) 4-chloroquinoline/MeOH/cat.HCl/ refluxes

N-(3-nitrophenyl)-4-pyridine amine (A3).(17.4g, 0.126mol) the suspension azeotropic in benzene is 10 hours for the tosic acid monohydrate.Add phenol (50g) and made the mixture azeotropic 2 hours, add then 4-pyridyl pyridinium chloride (26.6g, 0.138mmol) and the 3-N-methyl-p-nitroaniline (17.4g, 0.126mmol) and made the mixture azeotropic 3 hours.Benzene decompression is removed and the black residue that obtains is heated to 180 ℃, kept 1 hour.Reaction mixture is cooled to 20 ℃, and adds 4N NaOH (150mL).Mixture was stirred 30 minutes water (2L) dilution then.With this mixture at CH ₂Cl ₂Stir (1L) and the throw out that obtains is filtered and use more CH ₂Cl ₂Washing.Solid is from MeOH/H ₂The O recrystallization obtains A3 (11.9g), is yellow solid: mp (MeOH/H ₂O) 182-184 ℃.From CH ₂Cl ₂Washing lotion reclaims other material, uses Na ₂SO ₄Dry and be evaporated to drying.Resistates is dissolved in CH ₂Cl ₂(100mL) and add hexane (200mL), mixture was stirred 16 hours.The throw out that obtains filtered and with the ether washing removing any unreacted 3-N-methyl-p-nitroaniline, and from MeOH/H ₂O crystalline solid obtains other A3 (3.2g; Total recovery 15.7g, 58%); ¹H NMR[(CD ₃) ₂SO] δ 9.28 (s, 1H, NH), and 8.31-8.29 (m, 2H, H-2,6), 7.95 (t, J=2.1Hz, 1H, H-2 '), 7.82-7.79 (m, 1H, H-4 '), 7.65-7.57 (m, 2H, H-5 ', H-6 '), 7.03-7.02 (m, 2H, H-3, H-5), ¹³CNMR[(CD ₃) ₂SO] δ 150.5 (2C), 148.8,148.6,142.2,130.8,124.8,116.2,112.7,110.2 (2 * C); Ultimate analysis, C ₁₁H ₉N ₃O ₂: C, 61.4; H, 4.2; N, 19.5; Measured value: C, 64.5; H, 4.3; N.19.7%.

N ¹-(4-pyridyl)-1,3-phenylenediamine (A4).(7.26g 33.7mmol) and the suspension hydrogenation of 10%Pd/C in MeOH 2 hours, filters by diatomite (Celite) pad, and with solvent evaporation with compound A-13.Resistates is from MeOH/H ₂The O recrystallization obtains A4, is buff powder (5.43g, 83%): mp. (MeOH/H ₂O) 170-171 ℃. ¹HNMR[(CD ₃) ₂SO] and δ 8.48 (bs, 1H, NH), 8.14-8.12 (m, 2H, H-2 ﹠amp; 6), 6.96 (t, J=7.9Hz, 1H, H-5), 6.86-6.84 (m, 2H, H-3 ﹠amp; 6), 6.43 (t, J=2.0Hz, 1H, H-2), 6.32 (dd, J=7.8,1.3H, 1H, H-6), 6.26 (dd, J=7.8,1.5Hz, 1H, H-4) 5.07 (br s, 2H, NH ₂); HRMS (EI ⁺) calculated value, C ₁₁H ₁₁N ₃(M ⁺) m/z185.0953, measured value 185.0945; Ultimate analysis calculated value C ₁₁H ₁₁N ₃.0.125H ₂O:C, 70.5; H, 6.1; N, 22.5; Measured value: C, 70.6; H, 6.0; N, 22.7%.

N-(4-nitrophenyl)-3-(4-pyridinylamino) benzamide (A5).(4.3g 16.36mmol) is suspended in SOCl with the 4-nitrobenzoic acid ₂(30mL), add 2 DMF, and with 1 hour (up to obtaining clear soln) of mixture backflow.With the reaction mixture cool to room temperature and with excessive SOCl ₂Under vacuum, remove.The resistates that obtains is dissolved in 1,4-diox and join A4 (3.0g 16.20mmol) is comprising 1 of pyridine (8mL), in the suspension in the 4-diox (300mL).Reaction mixture was stirred 16 hours at 50 ℃, then with solvent evaporation.Resistates stirred in weak ammonia and the throw out that obtains filtered and, obtain A5 (5.3g, 79%): 219-222 ℃ of mp (MeOH) from the MeOH crystallization; ¹H NMR[(CD ₃) ₂SO] δ 10.63 (s, 1H, NH), 9.04 (s, 1H, NH), 8.37 (d, J=8.9Hz, 2H, ArH), 8.24-8.18 (m, 4H, ArH), 7.81 (bs, 1H, ArH), 7.45 (d, J=8.5Hz, 1H, ArH), 7.34 (t, J=8.0Hz, 1H, ArH), 6.99-6.95 (m, 3H, ArH) ¹³C NMR[(CD ₃) ₂SO] δ 166.9,164.0,150.3,149.4 (2 * C), 149.1,140.5,139.6,129.5 (2 * C), 129.2,123.5 (2 * C), 123.4,115.8,114.6,111.6,109; HRMS (FAB ⁺) calculated value C ₁₈H ₁₅N ₄O ₃(M ^{+ 1}) m/z 335.1144, measured value 335.1154.

1-methyl-4-{3-[(4-oil of mirbane amido) carbonyl] anilino } pyridinium chloride (A6).To A5 (507mg, DMF 1.51mmol) (2mL) solution add methyl tosylate (3mL), and with reaction mixture stirring at room 20 hours.Solvent decompression is removed and resistates is dissolved in MeOH again.With this solution evaporation to dry, and with resistates from the MeOH/EtOAc crystallization, obtain A6, be tosylate (530mg, 67%).Followingly be translated into chloride salt by ion-exchange.With AG ^R1-X ₄Resin 200-400 chloride form (7g) washes and adorns post with water.Tosylate A6 (530mg) stirs in through the resin (2g) of washing in advance, and the soup compound that obtains is loaded on the pillar.Merge and evaporate to dryness then with pillar water wash-out, and with the fraction of inclusion compound.(3 * 20mL, and finally from the MeOH/EtOAc redeposition, obtain A6 is chloride salt (317mg 54%): 295-298 ℃ of mp (MeOH/EtOAc) to make resistates and MeOH azeotropic. ¹H NMR[(CD ₃) ₂SO] δ 10.82 (s, 1H, NH), 10.76 (s, 1H, NH), 8.38 (d, J=8.9Hz, 2H, ArH), 8.31 (d, J=7.5Hz, 2H, ArH), 8.21 (d, J=8.9Hz, 2H, ArH), 7.96 (t, J=1.9Hz, 1H, ArH), 7.64 (dd, J=8.7,1.1Hz, 1H, ArH), 7.50 (t, J=5.4Hz, 1H, ArH), 7.22 (d, J=7.6Hz, 2H, ArH), 7.11 (dd, J=7.9,1.3Hz, 1H, ArH), 3.97 (s, 3H, N ⁺CH ₃); HRMS (FAB ⁺) calculated value C ₁₉H ₁₇N ₄O ₃(M ^{+ 1}) m/z 349.1301, measured value 349.1303.

The 4-{3-[(4-amino benzoyl) amino] anilino }-1-picoline dichloride (A7).(1.57g 4.08mmol) is dissolved in 5:1 H with A6 ₂O:EtOH (62mL) adds Fe powder (1.1g), and the suspension that obtains was refluxed 5 hours under vigorous stirring.The reaction mixture of heat is filtered by Celite pad, and with the EtOH washing of Celite pad with heat.The EtOH fraction that merges is evaporated to drying, and with the resistates warm water extraction.With this solution evaporation to dry, and by with methanol azeotropic (3 * 30mL) dried residue.Resistates is dissolved in MeOH (20mL), add hydrochloric acid methanol (1.25M, 5mL), and with solution stirring 30 minutes.With solution evaporation to dry, by with MeOH (3 * 30mL) azeotropic drying resistatess.Resistates obtains A7 (913mg, 63%) at last from the MeOH/EtOAc crystallization, is 269-273 ℃ of white solid: mp (MeOH/EtOAc); ¹H NMR[(CD ₃) ₂SO] δ 10.76 (s, 1H, NH), 10.05 (s, 1H, NH), 8.29 (d, J=7.4Hz, 2H, ArH), 7.95 (t, J=1.9Hz, 1H, ArH), 7.78 (d, J=8.7Hz, 2H, ArH), 7.60 (dd, J=8.7,1.1Hz, 1H, ArH), 7.60 (t, J=8.1Hz, 1H ArH), 7.20 (d, J=7.5Hz, 2H, ArH), 7.00 (dd, J=7.9,1.4Hz, 1H, ArH), 6.72 (d, J=8.6Hz, 2H, ArH), 3.96 (s, 3H, N ⁺CH ₃) do not observe NH ₂Signal; HRMS (FAB ⁺) calculated value C ₁₉H ₁₉N ₄O (M ⁺) m/z 319.1559, measured value 319.1562.Analyze CRL11720, calculated value C ₁₉H ₂₀N ₄Cl ₂O:C, 58.3; H, 5.4; N, 14.3; Measured value: C, 58.7; H, 5.3; N, 14.5%.

1-methyl-4-(3-{[4-(4-quinolyl amino) benzoyl] amino } anilino) pyridinium chloride (compd A).To A7 (200mg, 0.51mmol) suspension in MeOH (20mL) add the 4-chloroquinoline (100mg, 0.61mmol), and with mixture heating up 1 hour (up to obtaining clear soln) that reflux.Add a dense HCl then, and continue to reflux other 20 hours.Reaction mixture is evaporated to drying, and resistates is dissolved in MeOH (10mL).Add EtOAc (50mL) then, and the MeOH boiling is removed.The throw out that obtains is filtered,, and, obtain compd A (214mg, 81%), be light yellow solid: mp 253-257 ℃ from the MeOH/EtOAc crystallization with the EtOAc washing; ¹H NMR[(CD ₃) ₂SO] δ 11.07 (br, 1H, NH), 10.79 (s, 1H, NH), 10.60 (s, 1H, NH), 8.84 (d, J=13.5Hz, 1H, ArH), 8.61 (d, J=6.9Hz, 1H, ArH), 8.31 (d, J=7.4Hz, 2H, ArH), 8.18 (d, J=8.6Hz, 2H, ArH), and 8.12-8.04 (m, 2H, ArH), 7.99 (brs, 1H, ArH), 7.84 (t, J=7.2Hz, 1H, ArH), 7.68 (d, J=8.5Hz, 2H, ArH), 7.49 (t, J=8.1Hz, 1H, ArH), 7.23 (d, J=7.5Hz, 2H, ArH), 7.10 (brd, J=7.8Hz, 1H, ArH), 7.01 (d, J=6.7Hz, 1H, ArH), 3.98 (s, 3H, N ⁺CH ₃ ¹³C NMR[(CD ₃) ₂SO] δ 164.8,154.6,154.3,144.2,142.8,140.5,140.3,138.3,137.3,133.8,132.5,129.8,129.4 (2 * C), 127.0,124.4 (2 * C), 124.1,120.2,118.0,117.9,117.5,114.5,109.2,100.3,44.6, be difficult to observe the enhancing of two carbon signals; HRMS (FAB ⁺) calculated value C ₂₈H ₂₄N ₅O 446.1981, measured value 446.1975; Ultimate analysis, calculated value C ₂₈H ₂₅Cl ₂N ₅O.2.25H ₂O:C, 60.2; H, 5.3; N, 12.5; Measured value: C, 60.1; H, 5.3; N, 12.5%.

Embodiment B.

The preparation of 1-methyl-4-(4-(4-(quinolyl-4 amino) benzamido) phenyl amino) pyridinium chloride hydrochloride (compd B)

(i) TsOH/180 ℃; (ii) H ₂/ Pd/C/MeOH; (iii) 4-nitrobenzoyl chloride/pyridine/diox; (iv) DMF/MeOTs/20 ℃, ion-exchange then; (v) Fe powder/EtOH/H ₂O; (vi) 4-chloroquinoline/MeOH/cat.HCl/ refluxes

The N-4-nitrophenyl)-4-pyridine amine (B3).(53.80g 0.28mol) is dissolved in benzene (200mL) and the solution that obtains refluxed about 96 hours, up to H under the Dean-Stark condition with the 4-toluenesulphonic acids ₂The output of O stops.(112.25g 1.19mol), and refluxed the mixture that obtains about 1 hour, up to H under the Dean-Stark condition to add phenol to this solution ₂The output of O stops.After this, (59.95g, 0.31mol) (A2) and 4-N-methyl-p-nitroaniline (B1), and the mixture that obtains refluxed under the Dean-Stark condition about 2 hours are up to H to add 1-(4-pyridyl)-pyridinium chloride ₂The output of O stops.After this, the benzene decompression is removed, and the black residue that obtains is heated to 180 ℃, kept 2 hours.Fully alkalize then with the resistates cool to room temperature, and by adding the 4N NaOH aqueous solution.The solution H that obtains ₂O and CH ₂Cl ₂Dilution, and stirring at room 2 hours.Make the suspension filtered that obtains by Celite pad, obtain first N-methyl-p-nitroaniline B3 (0.22g), be meticulous yellow solid.Filtrate is diluted with MeOH, alkalizes with 4N NaOH again, extracts (x4) with EtOAc then.The organic extraction H that merges ₂O (x1), salt water washing, and use MgSO ₄Dry.The solvent decompression is removed, obtain second crowd of B3 (55g); ¹H NMR[(CD ₃) ₂SO]: δ 7.16 (dd, J=4.76,1.60Hz, 2H, ArH), 7.33 (ddd, J=10.37,5.32,3.23Hz, 2H, ArH), 8.18 (ddd, J=10.37,5.32,3.23Hz, 2H, ArH), 8.38 (dd, J=4.72,1.56Hz, 2H, ArH), 9.71 (br s, Ar-NH-Ar).LCMS(APCI ⁺)：216(100％)。

N ¹-(4-pyridyl)-1,4-phenylenediamine (B4).Make N-methyl-p-nitroaniline B3 (～55.00g ,～0.26mol) at 2:1 EtOH:H ₂Reflux among the O (500mL), add then the Fe powder (56.96g, 1.02mmol) and Glacial acetic acid (10mL).The mixture that obtains was refluxed 30 minutes, then cool to room temperature.By adding NH ₃The aqueous solution alkalizes reaction mixture and filters to remove solid.The solvent decompression is removed, obtain B4, be fluffy crystalline state magenta solid (28.49g, 54% from B1); ¹H NMR[(CD ₃) ₂SO]: δ 4.99 (brs, 2H, Ar-NH ₂), 6.58 (m, 4H, ArH), 6.86 (ddd, J=9.66,4.99,3.04Hz, 2H, ArH), 8.03 (d, J=6.28Hz, 2H, ArH), 8.24 (s, 1H, Ar-NH-Ar).LCMS(APCI ⁺)：186(100％)。

N-(4-nitrophenyl)-4-(4-pyridinylamino) benzamide (B5).To B4 (10.07g, 54.37mmol) at anhydrous pyridine (21.90mL, 271.82mmol) in solution add the 4-nitrobenzoyl chloride (10.09g, the 54.37mmol) solution in no Shui diox (100mL), and with the mixture that obtains 50 ℃ of heating 2 hours.After this, with the reaction mixture cool to room temperature and by adding NH ₃Aqueous solution alkalization.Collect the throw out that obtains by filtering, obtain first acid amides B5 (3.47g, 19%), be unbodied orange/yellow solid.Filtrate is extracted (x4) with EtOAc, and with the organic extraction salt water washing that merges, is evaporated to drying then, obtain B4 and B5 mixture (by ¹H NMR analyzes and is about 1:1).This mixture was handled 12 hours at 50 ℃ with the 4-nitrobenzoyl chloride once more, obtained other 4g B5; ¹H NMR[(CD ₃) ₂SO]: δ 6.86 (dd, J=4.82,1.60Hz, 2H, ArH), 7.21 (dt, J=9.86,5.02,3.00Hz, 2H, ArH), 7.76 (d, J=8.84,2H, ArH), 8.18 (m, 4H, ArH), 8.37 (ddd, J=9,23,4.33,2.34Hz, 2H, ArH), 8.73 (s, 1H, ArNHAr), 10.52 (s, 1H, ArC (O) NHAr).LCMS(APCI ⁺)：335(100％)。

1-methyl-4-{4-[(4-oil of mirbane amido) carbonyl] anilino } pyridinium chloride (B6).To acid amides B5 (5.62g, dry DMF 16.82mmol) (110mL) solution add the toluenesulphonic acids methyl esters (33.00mL, 218.68mmol) and with the mixture that obtains stirring at room 12 hours.After this, reaction mixture is filtered by Celite pad, obtain first tosylate of 11, be unbodied glassy yellow solid (7.57g, 86%).Filtrate decompression is concentrated,, and it is passed through filtration collect, obtain the tosylate (0.72g, 8%) of second crowd of B6 by Celite pad up to the formation throw out. ¹H?NMR[(CD ₃) ₂SO]：δ?2.29(s，3H，-O(O) ₂SPhCH ₃)，3.96(s，3H，R ₂N ⁺-CH ₃)，7.10(m，4H，O(O) ₂SArHCH ₃)，7.35(d，J＝8.84Hz，2H，ArH)，7.47(d，J＝8.08Hz，2H，ArH)，7.91(d，J＝8.86，2H，ArH)，8.20(dd，J＝6.94，1.94，ArH)，8.26(d，J＝7.45Hz，2H，ArH)，8.39(m，2H，ArH)，10.42(s，1H，ArNHAr)，10.70(s，1H，ArNHAr)。(APCI ⁺)：349(100％)。

(8.30g, 15.97mmol) (～suspension in 200mL) adds ion exchange resin, and (67g uses H in advance at MeOH to the tosylate of B6 ₂The O washing) [1-X ₄, BioRadAG, 200-400 chloride form].The suspension that obtains is loaded into more the polyion exchange resin, and (70g uses H in advance ₂The O washing) on the pillar and with pillar MeOH wash-out.The solvent decompression is removed, obtain the chloride salt (6.47g, 91%) of B6, be unbodied yellow solid; ¹H NMR[(CD ₃) ₂SO]: δ 3.95 (s, 3H, R ₂N ⁺-CH ₃), 7.10 (m, 2H, ArH), 7.35 (d, J=7.93Hz, 2H, ArH), 7.91 (d, J=8.52Hz, 2H, ArH), 8.22 (m, 4H, ArH), 8.39 (d, J=8.76Hz, 2H, ArH), 10.50 (brs, 1H, ArNHAr), 10.72 (s, 1H, ArNHAr).LCMS(APCI ⁺)：349(100％)。

The 4-{4-[(4-amino benzoyl) amino] anilino }-1-picoline dichloride (B7).(6.47g is 16.81mmol) at 2:1 EtOH:H to the acid amides B6 that is refluxing ₂Suspension among the O (600mL) sequentially add the Fe powder (3.76g, 67.24mmol) and Glacial acetic acid (12mL).The mixture that obtains was refluxed 2 hours, the reaction mixture of heat is filtered pass through Celite pad then.Solvent decompression is removed, and resistates is dissolved in MeOH again, and removal of solvent under reduced pressure once more.The processing of this back repeats twice again, several hydrochloric acid methanols of adding when dissolving again the last time (～4M).The solvent decompression is removed, and make resistates, obtain amine B8 (6.35g), be colour of loess green-white solid, its not purified use from the MeOH:EtOAc recrystallization. ¹H?NMR[(CD ₃) ₂SO]：δ?3.94(s，3H，R ₂N ⁺-CH ₃)，6.85(d，J＝8.26Hz，2H，ArH)，7.12(d，J＝7.12Hz，2h，ArH)，7.29(m，2H，ArH)，7.83(d，J＝8.64Hz，2H，ArH)，7.89(m，2H，ArH)，8.24(d，J＝7.42Hz，2H，ArH)，10.10(s，1H，ArNHAr，10.73(s，1H，ArH)。LCMS(APCI ⁺)：319(100％)。

1-methyl-4-[4-(4-[(6-nitro-4-quinolyl) and amino] benzoyl } amino) anilino] pyridinium chloride (compd B).With the 4-chloroquinoline (89mg, 0.55mmol) and 3 dense HCl add sequentially that (214mg 0.55mmol) in the solution in anhydrous MeOH (19mL), makes the mixture that obtains reflux 31 hours then to amine B7.After this, the solvent decompression is removed, and circulate the resistates drying that obtains by two MeOH-azeotropic.Resistates is further purified by preparation property HPLC then from MeOH:EtOAc crystallization twice, obtains compd B, is unbodied yellow solid (68mg, 24%). ¹H NMR[(CD ₃) ₂SO]: 3.96[s, 3H, ArN ⁺CH ₃], 7.00[d, J=6.92Hz, 1H, ArH], 7.14[d, J=7.29Hz, 2H, ArH], 7.35[d, J=8.86Hz, 2H, ArH] and, 7.69[d, J=8.58Hz, 2H, ArH], 7.85[t, J=15.42,7.71Hz, 1H, ArH], 7.96[d, J=8.86,2H, ArH], 8.07[t, J=15.42,7.71Hz, 1H, ArH], 8.13[d, J=7.76Hz, 1H, ArH], 8.20[d, J=8.58Hz, 2H, ArH], 8.26[d, J=7.39Hz, 2H, ArH], 8.61[d, J=6.92Hz, 1H, ArH], 8.88[d, J=8.43Hz, 1H, ArH], 10.58[s, 1H, ArNHAr], 10.76[s, 1H, ArNHAr], 11.16[s, 1H, ArC (O) NHAr], 14.81[brs, quinoline-N ⁺H] .LCMS (APCI ⁺): 447 (100%).HPLC：99.7％。

Embodiment C.

4-[4-({ 3-[(1-methyl-4-pyridyl) amino] benzoyl } amino) anilino] preparation of quinoline dichloride (Compound C)

Reagent: (i) CDl/DMF/55 ℃ of 1h, 4-N-methyl-p-nitroaniline/DMAP/150 ℃ then; (ii) p-TsOH; (iii) pyridyl pyridinium chloride/180 ℃; (iv) DMF/MeOTs; (v) Fe powder/EtOH/H ₂O/H ⁺/ ion-exchange then; (vi) 4-chloroquinoline/EtOH/H ₂O/H ⁺/ reflux

(acetylamino)-N-(4-nitrophenyl) benzamide (C2).With 3-acetaminobenzoic acid (C1) (5.30g, 29.58mmol) and the mixture of CDI (5.75g, 35.50mmol, 1.2 equivalents) in N-Methyl pyrrolidone (20mL) 55-60 ℃ the heating 2 hours.Add 4-N-methyl-p-nitroaniline (6.13g, 44.37mmol, 1.5 equivalents) then and, kept 4 hours, be poured in the water (400mL) then and stirred 18 hours mixture heating up to 140 ℃.The throw out that obtains is filtered also sequentially water and CH ₂Cl ₂Washing then from the MeOH crystallization, obtains C2 (4.84g; 55%), is yellow solid; 259-261 ℃ of mp (MeOH); ¹H NMR[(CD ₃) ₂SO] δ 10.80 (s, 1H, NH), 10.15 (s, 1H, NH), 8.29 (m, 2H, H-3 ﹠amp; H-5), 8.12 (t, J=1.8Hz, 1H, H-2), 7.84 (br dd, J=8.1,1.2Hz, 1H, H-6), 7.65 (td, J=7.8,2.5Hz, 1H, H-4), 7.48 (t, J=7.9Hz, 1H, H-5), 2.08 (s, 3H, COCH ₃); Ultimate analysis calculated value C ₁₅H ₁₃N ₃O ₄: C, 60.2; H, 4.4, N, 14.0; Measured value C, 60.4; H, 4.6; N, 14.1%.

3-amino-N-(4-nitrophenyl) benzamide (C3).(4.25g 15.9mmol) is suspended in 1, in the 4-diox (100mL), adds rare HCl (dense HCl+85mL H of 15mL with Compound C 2 ₂O), and with mixture reflux 6 hours (up at 6% MeOH:CH ₂Cl ₂In TLC show that the starting raw material completely consumed falls).Then reaction mixture is cooled to 20 ℃ and with solvent evaporation to dry.The resistates that obtains is at rare NH ₃Stir in the aqueous solution, filter then, wash with water, oven drying and from the MeOH crystallization obtains amine C3 (2.75g, 67%); 229-232 ℃ of mp (MeOH); ¹H NMR[(CD ₃) ₂SO] and δ 10.63 (s, 1H, CONH), 8.27-8.22 (m, 2H, H-3 ′ ﹠amp; H-5 '), 8.06-8.02 (m, 2H, H-2 ′ ﹠amp; H-6 '), 7.18 (t, J=7.7Hz, 1H, H-5), 7.12-7.08 (m, 2H, H-2﹠amp; H-6), and 6.80-6.78 (m, 1H, H-4), 5.38 (s, 2H, NH ₂); HRMS (FAB ⁺) calculated value C ₁₃H ₁₁N ₃O ₃(M ^{+ 1}) m/z 258.0879, measured value 258.0875; Ultimate analysis calculated value C ₁₃H ₁₁N ₃O ₃: C, 60.7; H, 4.3; N, 16.3; Measured value: C, 60.7; H, 4.4; N, 16.6%.

N-(4-nitrophenyl)-3-(4-pyridinylamino) benzamide (C4).(2g is 10.49mmol) with benzene (250mL) azeotropic 2 hours to make tosic acid.Add pyridyl pyridinium chloride (3.3g) then, Compound C 3 (2.7g, 10.49mmol) and N-Methyl pyrrolidone (10mL) and with mixture 130 ℃ of azeotropic 2 hours.Solvent is seethed with excitement, and the dun mixture that obtains was heated 1 hour at 180 ℃, be cooled to about 50 ℃ and dilute with water then.Mixture stirring at room 2 hours, and is filtered the throw out that obtains and water sequentially rare NH ₃The aqueous solution and CH ₂Cl ₂Washing.Make solid from the MeOH crystallization then, filter and with MeOH and CH ₂Cl ₂Washing obtains C4 (1.92g, 55%); ¹H NMR[(CD ₃) ₂SO] δ 10.79 (s, 1H, CONH), 9.00 (s, 1H, NH), 8.29-8.22 (m, 4H, H-3 ', 5 ′ ﹠amp; PyH-2,6)), 8.08-8.04 (m, 2H, H-2 ', 6 '), 7.76 (t, J=1.8Hz, 1H, H-2), 7.63 (td, J=8.0,1.3H, Hz, 1H, H-6), 7.52 (t, J=7.8Hz, 1H, H-5), 7.46-7.43 (m, 1H, H-4), 6.98-6.97 (m, 2H, pyH-3﹠amp; 5).

1-methyl-4-{3-[(4-oil of mirbane amido) carbonyl] anilino } pyridine 4-toluenesulfonate (C5).With Compound C 4 (1.92g 5.73mmol) is suspended among the DMF (10mL), add methyl tosylate (15mL) and with mixture stirring at room 18 hours.Removal of solvent under reduced pressure also is dissolved in warm MeOH (10mL) with resistates, dilutes with EtOAc then.Some MeOH are seethed with excitement, then with freezing 18 hours of solution.The throw out that obtains is filtered, and with the EtOAc washing, obtain pure basically C5 (2.90g, 97%), it is without being further purified use. ¹H?NMR[(CD ₃) ₂SO]δ?10.85(s，1H，CONH)，10.60(s，1H，NH)，8.33-8.27(m，4H，H-3′，5′&py?H-2，6)，8.08-8.30(m，2H，H-2′，6′)，7.94-7.89(m，2H，H-2＂4＂)，7.69(t，J＝7.9Hz，1H，H-5＂)，7.60-7.57(m，1H，H-6＂)，7.48-7.45(m，2H，H-3，5)，7.22(d，J＝7.5Hz，2H，p-tol?H-2，6)，7.20(d，J＝8.1Hz，2H，p-tolH-3，5)，3.99(s，3H，N ⁺CH ₃)，1.99(s，3H，CH ₃)。

4-{3-[(4-amino-benzene amido) carbonyl] anilino }-1-picoline muriate (C6).(1.59g 3.06mmol) is dissolved in～6:1 EtOH:H with Compound C 5 ₂Among the O (46mL), add Fe powder (885mg), and make the suspension returning that obtains.Add two dense HCl, and continue to reflux 1 hour (up to using N-BuOH:H ₂O:CH ₃CO ₂The TLC demonstration starting raw material completely consumed of carrying out mutually on the 5:4:1 mixture of H is fallen).Reaction mixture dilutes with EtOH (100mL) and makes its backflow.The reaction mixture of heat filtered by Celite pad and with the top layer of Celite pad seethe with excitement, and filter (this process triplicate is to guarantee fully to extract product from the iron resistates) with more EtOH.The EtOH extracting solution that merges is evaporated to drying and with the resistates hot water extraction, and Celite pad is passed through in filtration.The volume that the filtrate simmer down to is little stirs in the ion exchange resin (55g) of washing in advance then.Be carried on the pillar of filling the soup compound that obtains and the water wash-out with the ion exchange resin of washing in advance.The fraction that will comprise product merges and is evaporated to drying, and with resistates and EtOH azeotropic several times.Resistates is dissolved in the MeOH of a little volume, add hydrochloric acid methanol (1.25M, 1mL), and with solution stirring 10 minutes.Solution dilutes with EtOAc, and the throw out that obtains is filtered, and then from the MeOH/EtOAc crystallization, obtains C6 (873mg, 80%); Mp (MeOH/EtOAc)〉300 ℃; ¹H NMR[(CD ₃) ₂SO] δ 10.77 (brs, 1H, NH), 9.95 (s, 1H, NH), 8.41 (brd, J=5.5Hz, 2H, PyH-2,6), 7.87-7.84 (m, 2H, H-2,4), 7.61 (t, J=7.8Hz, 1H, H-5), 7.49 (brd, J=7.8Hz, 1H, H-6), 7.37 (d, J=8.7Hz, 2H, H-2 ', 6 '), (7.21 brd, J=4.6Hz, 2H pyH-3,5), 6.55 (d, J=8.8Hz, 2H, H-3 ', 5 '), 4.93 (s, 2H, NH ₂), 3.97 (s, 3H, N ⁺CH ₃).

4-[4-(3-[(1-methyl-4-pyridyl) and amino] benzoyl } amino) anilino] quinoline dichloride (Compound C).(100mg 0.28mmol) is dissolved in EtOH (10mL) and H with Compound C 6 by heating ₂O (5mL).(60mg 0.36mmol) with 3 dense HCl, and makes mixture reflux 18 hours (up to using N-BuOH:H to add the 4-chloroquinoline ₂O:CH ₃CO ₂The initial amine completely consumed of carrying out mutually on the 5:4:1 mixture of H of TLC demonstration is fallen).With the EtOAc dilution, the boiling several minutes makes its cool to room temperature to reaction mixture then then.The throw out that obtains is filtered and, obtain Compound C (64mg, 63%) from the MeOH/EtOAc crystallization; Mp (MeOH, EtOAc)〉300 ℃; ¹H NMR[(CD ₃) ₂SO] δ 14.49 (br, 1H, N ⁺H), 11.01 (s, 1H, NH), 11.78 (br, 1H, NH), 10.64 (s, 1H, NH), 8.77 (d, J=8.5Hz, 1H, ArH), 8.57 (d, J=6.5Hz, 1H, ArH), 8.32 (d, J=7.5Hz, 2H, ArH), 8.06-7.92 (m, 6H, ArH), 7.53 (t, J=7.7Hz, 1H, ArH), 7.70-7.65 (m, 1H, ArH), 7.48 (d, J=8.8Hz, 2H, ArH), 7.29 (d, J=7.5Hz, 2H, ArH), 6.79 (d, J=6.8Hz, 1H, ArH), 3.98 (s, 3H, N ⁺CH ₃).

Embodiment D.

4-[4-({ 4-[(1-methyl-4-pyridyl) amino] benzoyl } amino) anilino] preparation of quinoline dichloride (Compound D)

(i) TsOH/180 ℃; (ii) SOCl ₂, 4-N-methyl-p-nitroaniline/pyridine then; (iii) EDMF/MeOTs/20 ℃, ion-exchange then; (iv) Fe powder/EtOH/ trace HCl; (v) 4-chloroquinoline/EtOH/H2O/H+/backflow

4-(4-pyridinylamino) phenylformic acid (D2).(17.4g is 0.126mol) with benzene (300mL) azeotropic 5 hours to make tosic acid.Add then 4-pyridyl pyridinium chloride (26.6g, 0.138mol), 4-benzaminic acid (D1) (17.3g, 0.126mol) and N-Methyl pyrrolidone (40mL), and with mixture in oil bath 110 ℃ of azeotropic 1 hour.At 130 ℃ benzene is evaporated, raise the temperature to 180 ℃ then, kept 1.5 hours.Dilute with the reaction mixture cool to room temperature and with salt solution then.The mixture that obtains is warm, obtain throw out, it is filtered then at 110 ℃ of oven dryings.Then with solids extract in ebullient ethanol (5 * 150mL), and the EtOH extracting solution that merges is evaporated to drying.With the resistates that obtains at rare NH ₃Stir in the aqueous solution, and the throw out that obtains is filtered.By obtain the throw out that other product and filtration obtain with ice AcOH neutralization filtrate.With each crude product batch of material merging and from H ₂The O crystallization obtains D2 (8.8g, 32%): mp (H ₂O) 333-336 ℃; ¹H NMR[(CD3) 2SO] 12.05 (br., 1H, COOH), 9.17 (s, 1H, NH), 8.28-8.27 (m, 2H, ArH), 7.90-7.86 (m, 2H, ArH), 7.26-7.23 (m, 2H, ArH), 7.05-7.03 (m, 2H, ArH), ¹³C NMR[(CD ₃SO) ₂] δ 166.8,150.2 (2 * C), 148.6,145.0,130.9 (2 * C), 123.4,117.32 ((2 * C), 110.6 (2 * C); LCMS 215+ve.

N-(4-nitrophenyl)-4-(4-pyridinylamino) benzamide (D3).(172mg is 0.80mmol) at the SOCl that comprises catalytic amount DMF to make Compound D 2 ₂1 hour (up to obtaining clear soln) refluxes (5mL).With the reaction mixture cool to room temperature and with excessive SOCl ₂Under vacuum, remove.Add diox (10mL) to resistates, then its vacuum is removed.With resistates cooling in dry ice-propanone is bathed, (112mg 0.81mmol) and pyridine (1.3mL), adds Et subsequently to add p-Nitroaniline ₃N (0.3mL) and with mixture stirring at room 20 minutes, refluxed then 1 hour.With the reaction mixture cool to room temperature and use H ₂NH is used in the O dilution then ₃The aqueous solution alkalizes and stirred 30 minutes.The throw out that obtains is filtered, and water sequentially, hexane and CH ₂Cl ₂Washing.Crude product is dissolved in MeOH, is adsorbed onto on the silica gel then, and with the adsorptive (adsorbate) that obtains in the enterprising circumstances in which people get things ready for a trip of silica gel spectrumization, use 0-10%MeOH:CH ₂Cl ₂Wash-out obtains D3 (85mg, 32%); Mp 298-302 ℃ (MeOH); ¹H NMR[(CD ₃) ₂SO] δ 10.64 (s, 1H, NH), 9.21 (s, 1H, NH), 8.30-8.24 (m, 4H, ArH), 8.09-8.05 (m, 2H, ArH), 8.00-7.98 (m, 2H, ArH), 7.34-7.31 (m, 2H, ArH), 7.07-7.05 (d, m, 2H, ArH); LCMS 235+ve.

1-methyl-4-{4-[(4-oil of mirbane amido) carbonyl] anilino } pyridinium chloride (D4).With compound d3 (80mg 0.24mmol) is dissolved in DMF (0.8mL), adds methyl tosylate (0.5mL), and with mixture stirring at room 18 hours.With solvent removed under reduced pressure, and resistates is dissolved in MeOH (1mL), uses EtOAc (50mL) dilution then.The throw out that will form when cooling filters, and then from the MeOH/EtOAc crystallization, obtains D5 (119mg), is its tosylate.Followingly be translated into chloride salt by ion-exchange.With AG ^R1-X ₄Resin 200-400 chloride form (3g) washes with water and is filled in the pillar.Tosylate D5 (119mg) is stirred in the resin (1g) of washing in advance, and the soup compound that obtains is loaded on the pillar.Pillar is used 50%MeOH:H then ₂O, wash-out also will contain the fraction merging of compound and be evaporated to drying.Resistates and MeOH azeotropic (3 * 20mL), and, obtain D4 finally from the MeOH/EtOAc redeposition, be chloride salt (82mg, 88%); Mp (MeOH/EtOAc); ¹H NMR[(CD ₃) ₂SO] δ 11.00 (s, 1H, NH), 10.87 (s, 1H, NH), 8.37 (d, J=7.3Hz, 2H ArH), 8.30-8.26 (m, 2H, ArH), 8.15-8.08 (m, 4H, ArH), 7.52 (d, J=8.6Hz, 2H, ArH), 7.33 (d, J=7.2Hz, 2H, ArH), 4.01 (s, 3H, N ⁺CH ₃); LCMS 349+ve.

4-{4-[(4-amino-benzene amido) carbonyl] anilino }-1-picoline muriate (D5).(295mg 0.77mmol) is dissolved in～5:1 EtOH:H with Compound D 4 ₂O (12mL), and add Fe powder (209mg).With the mixture vigorous stirring and refluxed 4 hours, heat filtering passes through Celite pad then.Diatomite with ebullient EtOH washing, is merged the EtOH fraction and is evaporated to drying.Resistates is extracted in the hot water, and filter, be evaporated to drying then by diatomite.Resistates and MeOH azeotropic (3 * 20mL), be dissolved in then among the MeOH (10mL).Add hydrochloric acid methanol (1.25M, 5mL), and with solution stirring 10 minutes.Removal of solvent under reduced pressure and with resistates from the MeOH/EtOAc crystallization, obtain D5 (218mg, 80%); Mp (MeOH/EtOAc); ¹H NMR[(CD ₃) ₂SO] δ 11.13 (s, 1H, NH), 10.44 (s, 1H, NH), 9.88 (br, 2H, NH ₂), 8.36 (d, J=7.5Hz, 2H, ArH), 8.12 (d, J=8.6Hz, 2H, ArH), 7.86 (d, J=8.8Hz, 2H, ArH), 7.50 (d, J=8.8Hz, 2H, ArH), 7.32 (d, J=8.6Hz, 2H, ArH), 7.32 (d, J=7.5Hz, 2H, ArH), 7.30 (d, J=8.7Hz, 2H, ArH), 4.00 (s, 3H, N ⁺CH ₃); LCMS 319+ve

4-[4-(4-[(1-methyl-4-pyridyl) and amino] benzoyl } amino) anilino] quinoline dichloride (Compound D).(100mg 0.28mmol) is dissolved in EtOH (10mL) and H with Compound D 5 by heating ₂O (5mL).Add the 4-chloroquinoline (60mg, 0.36mmol) and dense HCl (3), and with reaction mixture refluxed 18 hours (up to using N-BuOH:H ₂O:CH ₃CO ₂The initial amine completely consumed of TLC demonstration of the last phase of the 5:4:1 mixture of H is fallen).Reaction mixture dilutes with EtOAc, and the backflow several minutes makes its cooling then.The throw out that obtains is filtered and, obtain Compound D (88mg, 63%) from the MeOH/EtOAc crystallization; Mp (MeOH, EtOAc)〉300 ℃; ¹H NMR[(CD ₃) ₂SO] δ 14.38 (br, 1H, N ⁺H), 11.04 (s, 1H, NH), 10.84 (br, 1H, NH), 10.53 (s, 1H, NH), 8.76 (d, J=8.6Hz, 1H, ArH), 8.58 (d, J=6.7Hz, 1H, ArH), 8.37 (d, J=7.5Hz, 2H, ArH), 8.14 (d, J=8.7Hz, 2H, ArH), 8.06-8.01 (m, 4H, ArH), 7.83-7.79 (m1H, ArH), 7.53-7.48 (m, 4H, ArH), 7.33 (d, J=7.6Hz, 2H, ArH), 6.79 (d, J=6.9Hz, 1H, ArH), 4.00 (s, 3H, N ⁺CH ₃).

Embodiment E.

The preparation of N-(4-(2-amino-6-methylpyrimidine-4-base is amino) phenyl)-4-(quinolyl-4 amino) benzamide hydrochloride salt (compd E)

6-methyl-N ⁴-(4-nitrophenyl) pyrimidine-2,4-diamines (E2).(9.30g 0.07mol) is dissolved in cellosolvo (330mL) with 4-N-methyl-p-nitroaniline (B1) [9.22g, 0.07mol] and 2-amino-4-chloro-6-methylpyrimidine (E1).Add several dense HCl to the solution that obtains, and the mixture that obtains was refluxed 30 minutes, its cool to room temperature is spent the night.After this, reaction mixture is filtered, and by adding ammonia soln with the solid alkalization that obtains, then from H ₂The O:EtOH crystallization obtains amine E2, is unbodied glassy yellow solid (7.33g).Filtrate is concentrated, alkalize as previously mentioned subsequently and redeposition, obtain other amount (0.78g, total recovery 51%). ¹H?NMR[(CD ₃) ₂SO]：2.14[s，3H，ArCH ₃]，6.00[s，1H，ArNHAr]，6.37[s，2H，ArNH ₂]，8.00[ddd，J＝10.24，5.05，2.96Hz，2H，ArH]，8.12[ddd，J＝10.24，4.95，2.90Hz，2H，ArH]，9.75[s，1H，ArH].LCMS(APCI ⁺)：246(100％)。

N ⁴-(4-aminophenyl)-6-methylpyrimidine-2,4-diamines (E3).To (5.46g is 0.02mol) at 2:1EtOH:H at the amine E2 that refluxes ₂Suspension among the O (100mL) sequentially add the Fe powder (4.97g, 0.09mol) and AcOH (2mL 2%v/v) and with the brown black suspension liquid that obtains refluxed～14 hours.After this, the reaction mixture filtration of heat is removed by Celite pad and with the solvent decompression.The resistates hot water extraction that obtains, and with waterborne suspension filtration the passing through Celite pad that obtains.The solvent decompression is removed, and resistates is used hot water extraction once more.With aq suspension filtration the passing through Celite pad that obtains.Remove and desolvate, obtain amine E3, be brown-white crystalline solid (4.85g, quantitative yield). ¹H?NMR[CD ₃) ₂SO]：2.01[s，3H，ArCH ₃]，5.68[s，1H，ArNHAr]，5.88[brs，2H，ArNH ₂]，6.51[ddd，J＝9.67，4.85，2.94Hz，2H，ArH]，7.14[d，J＝8.52Hz，2H，ArH]，8.35[s，1H，ArH].LCMS(APCI ⁺)：216(100％)。

N-[4-(2-amino-6-methylpyrimidine-4-base is amino) phenyl]-4-nitrobenzamide (E4).With the 4-nitrobenzoyl chloride (11.00g, 0.06mol) and anhydrous pyridine (9.08mL, 0.11mol) sequentially join amine E3 (4.85g, in no Shui diox (200mL) solution 0.02mol), and with the solution that obtains～50 ℃ of heating 5 days.After this, with the reaction mixture cool to room temperature, then by adding the ammonia soln alkalization.The suspension filtered that obtains is passed through Celite pad, obtain acid amides E4, be unbodied yellow solid (3.40g, 41%). ¹HNMR[(CD ₃) ₂SO]：2.09[s，3H，ArCH ₃]，5.87[s，1H，ArNHAr]，6.08[brs，6.08，ArNH ₂]，7.68[m，4H，ArH]，8.18[d，J＝8.80Hz，2H，ArH]，8.36[d，J＝8.80Hz，2H，ArH]，8.95[s，1H，ArH]，10.45[s，1H，ArC(O)NHAr].LCMS(APCI ⁺)：365(100％)。

4-amino-N-[4-(2-amino-6-methylpyrimidine-4-base is amino) phenyl] benzamide hydrochloride salt (E5).To (2.10g is 5.77mmol) at 2:1EtOH:H at the acid amides E4 that refluxes ₂Suspension among the O (100mL) sequentially adds the Fe powder, and (1.29g 23.09mol) and dense HCl (1-2mL), and refluxes the mixture that obtains 24 hours.After this, Celite pad is passed through in the reaction mixture filtration of heat, and the solvent decompression is removed.The resistates that obtains is from MeOH: the EtOAc crystallization, obtain amine E5 (being two batch of materials), and be cream-colored amorphous solid (1.94g, 90%). ¹H?NMR[(CD ₃) ₂SO])：2.27[s，3H，ArCH ₃]，6.16[brs，2H，ArNH ₂]，6.75[d，J＝8.36Hz，2H，ArNH ₂]，7.74[m，8H，ArH]，9.90[brs，1H?ArH]，10.55[brs，1H，ArNHAr]，12.58[brs，1H，ArC(O)NH].LCMS(APCI ⁺)：335(100％)。

N-(4-(2-amino-6-methylpyrimidine-4-base is amino) phenyl)-4-(quinolyl-4 amino) benzamide hydrochloride salt (compd E).(267mg is 0.720mmol) at～1:1:1MeOH:EtOH:H to amine E5 ₂(1.06g 6.48mmol) and 3 dense HCl, and refluxes the mixture that obtains～24 hours sequentially to add the 4-chloroquinoline in the solution among the O.After this, solvent decompression is removed and by two MeOH-azeotropic circulations with the resistates drying that obtains.Resistates is further purified by preparation property HPLC then from the MeOH/EtOAc redeposition, obtains compd E, is unbodied yellow solid (16mg, 4%). ¹H NMR[(CD ₃) ₂SO]: 1.91[s, 3H, ArCH ₃], 6.11[s, 1H, ArH], 7.02[d, J=6.77Hz, 1H, ArH], 7.65[d, J=8.55Hz, 2H, ArH] and, 7.71[brs, 2H, ArNH ₂], 7.84[m, 4H, ArH], 8.04[m, 2H, ArH], 8.15[d, J=8.55Hz, 2H, ArH] and, 8.63[d, J=6.77Hz, 1H, ArH], 8.70[d, J=8.50Hz, 1H, ArH] and, 10.38[s, 2H, ArNHAr ﹠amp; ArH], 10.77[brs, 1H, ArNHAr], 13.10[vvbrs, 2H, ArC (O) NHAr ﹠amp; Quinoline-N ⁺H] .LCMS (APCI ⁺): 463 (100%).HPLC：95.7％。

Embodiment F.

The preparation of 1-methyl-4-(3-{[4-(6-nitro-4-quinolyl amino) benzoyl] amino } anilino) pyridinium chloride (compound F 17-hydroxy-corticosterone)

(i) MeOH/cat.HCl/ refluxes

Make A7 and 4-chloro-6-nitroquinoline (F1) [Simpso N ﹠amp as mentioned above; Wright, J.Chem.Soc., 1948,1707] coupling, obtain compound F 17-hydroxy-corticosterone, 97% yield: mp273-277 ℃; ¹H NMR[(CD ₃) ₂SO] δ 11.33 (br, 1H, NH), 10.70 (s, 1H, NH), 10.60 (s, 1H, NH), 8.72-8.69 (m, 2H, ArH), 8.31 (d, J=7.4Hz, 2H, ArH), 8.24 (d, J=9.3Hz, 1H, ArH), 8.18 (d, J=8.5Hz, 2H, ArH), 7,98 (brs, 1H, ArH), 7.68-7.65 (m, 3H, ArH), 7.49 (t, J=8.1Hz, 1H, ArH), 7.21 (d, J=7.5Hz, 2H, ArH), and 7.12-7.01 (m, 2H, ArH), 4.02 (s, 3H, N ⁺CH ₃); HRMS (FAB) calculated value C ₂₈H ₂₃N ₆O ₃(M ⁺) m/z491.1832, measured value 491.1822.

Embodiment G.

The preparation of 1-methyl-4-(4-(4-(6-nitroquinoline-4-base is amino) benzamido) phenyl amino) pyridinium chloride hydrochloride (compound G)

(i) MeOH/cat.HCl/ refluxes

(2.09g, 5.32mmol) suspension in anhydrous MeOH (100mL) sequentially adds 4-chloro-6-nitroquinoline (F1) [1.11g, 5.32mmol] and several dense HCl, and the mixture that obtains was refluxed 12 hours to acid amides B7.After this, removal of solvent under reduced pressure, and with the resistates resuspending in 1:1 MeOH:EtOAc.Heat this mixture to remove MeOH, cooling then, and by filtering the throw out that collection obtains.Make this solid from the MeOH:EtOH recrystallization, obtain nitroquinoline compound G (2.48g, 89%), be unbodied yellow solid; ¹H NMR[CD ₃) ₂SO]: δ 3.96 (s, 3H, R ₂N ⁺-CH ₃), 7.12 (m, 3H, ArH), 7.35 (d, J=8.81Hz, 2H, ArH), 7.68 (d, J=8.51Hz, 2H, ArH), (7.96 d, J=8.81Hz, 2H ArH), 8.24 (m, 5H, ArH), 8.72 (m, 2H, ArH), 9.83 (d, J=1.69Hz, 1H, ArH), 10.56 (s, 1H, ArNHAr), 10.67 (s, 1H, ArNHAr), 11.48 (brs, 1H, ArNHAr).LCMS(APCI ⁺)：492(20％)。

Embodiment H.

The preparation of N-(4-(2-amino-6-methylpyrimidine-4-base is amino) phenyl)-4-(6-nitroquinoline-4-base is amino) benzamide hydrochloride salt (compound H)

(i)MeOH，cat.c.HCl，2.5d

N-[4-(2-amino-6-methylpyrimidine-4-base is amino) phenyl]-4-(6-nitroquinoline-4-base is amino) benzamide hydrochloride salt (compound H).To amine E5 (304mg, 0.82mmol) solution in anhydrous MeOH (30mL) sequentially add 4-chloro-6-nitroquinoline (171mg, 0.82mmol) and dense HCl (c.HCl) (1-2 drips) and the mixture that obtains refluxed～12 hours.MS afterwards and TLC analyze to be presented at and still have some 6 to exist in the reaction mixture, therefore at 12 hours with added the E5 of other five equilibrium in 24 hours (171mg is 0.82mmol) with dense HCl (1-2 drips).Then solvent is removed from reaction mixture, and the resistates that obtains is dissolved in MeOH, and then concentrating under reduced pressure.Resistates is through the circulation of another MeOH azeotropic, then with the resistates that obtains from MeOH:Et ₂The O redeposition obtains compound H, is unbodied yellow solid (383mg, 86%). ¹H?NMR(400MHz，DMSO)：2.28[s，3H，ArCH ₃]，6.18[brs，1H，ArH]，7.11[d，J＝6.59Hz，1H，ArH]，7.65Hz[d，J＝8.52Hz，2H，ArH]，7.81[m，5H，ArH?&?ArNH ₂]，8.17[d，J＝8.52Hz，2H，ArH]，8.25[d，J＝9.30Hz，1H，ArH]，8.69[m，2H，ArH]，9.77[s，1H，ArH]，10.42[s，1H，ArH]，10.62[brs，1H，ArNHAr]，11.20[v?brs，1H，ArNHAr]，12.63[v?brs，1H，ArC(O)NH].LCMS(APCI ⁺)：508(100％)。HPLC：98.5％。

Example I.

(E)-and N-[4-{1-([diamino methene base] hydrazono-) ethyl } phenyl]-preparation of 4-(6-nitroquinoline-4-base amino) benzamide hydrochloride salt (Compound I)

(E)-N-[4-(1-{ diamino methene base } hydrazono-) ethyl]-4-oil of mirbane hydrochloride (I3).With 4-nitro-acetophenone (I1) (49.95g, 0.30mol), aminoguanidine sulfate (I2) (51.77g, 0.20mol) and dense HCl (10mL 0.33mol) is incorporated among the MeOH (600mL), and the mixture that obtains refluxed 1 hour, its cool to room temperature is spent the night.After this, removal of solvent under reduced pressure, and by filtering the collection resistates.The solid that obtains is sequentially used MeOH and hexane wash, obtain diamines I3, be unbodied white solid (88.19g, quantitative yield) that it is without using through single step purification. ¹H?NMR[CD ₃) ₂SO)：2.36[s，3HArC(CH ₃)＝N-]，7.72[brs，4H，＝C(NH ₂) ₂]，8.23[m.5H，ArH]，10.42[v?brs，1.5H].LCMS(APCI ⁺)：222(100％)。

(E)-N-[4-(1-{ diamino methene base } hydrazono-) ethyl]-4-aniline dihydrochloride (I4).With the Fe powder (35.00g, 0.62mol) and dense HCl (4mL) join sequentially that (40.00g is 0.16mmol) at 2:1 EtOH:H at the diamines I3 that refluxes ₂In the suspension among the O (200mL), and the yellow suspension that obtains refluxed 14 hours.After this, Celite pad is passed through in the reaction mixture filtration of heat, and the solvent decompression is removed, obtain triamine I4, be Vandyke brown resin-like glassy mass (34.93g, 85%).This material is without being further purified use. ¹H?NMR[CD ₃) ₂SO)：2.20[s，3H?ArC(CH ₃)＝N-]，5.48[brs，2H，ArNH ₂]，5.54[d，J＝7.98Hz，2H，ArH]，7.58[v?br?s，4H，＝C(NH ₂) ₂]，7.63[d，J＝7.98Hz，ArH]，10.83[br?s，1H，＝N ⁺(H)-N＝].LCMS(APCI ⁺)：192(100％)。

(E)-N-[4-(1-{[diamino methene base] hydrazono-} ethyl) phenyl]-4-nitrobenzamide hydrochloride (I5).With 4-nitrobenzoyl chloride (16.01g, 86.26mmol) join triamine I4 (8.59g, 32.51mmol) and anhydrous pyridine (13.09mL is 162.53mmol) in the solution in no Shui diox (300mL), and made the suspension returning that obtains 2 hours, its cool to room temperature is spent the night.After this, reaction mixture is filtered, obtain solid, make it, obtain the acid amides I5 of three batch of materials, be unbodied cream color-light yellow solid (adding up to 3.50g, 29%) from the MeOH-EtOAc crystallization. ¹H?NMR[CD ₃) ₂SO)：2.34[s，3H，ArC(CH ₃)＝N-]，7.73[brs，4H，＝C(NH ₂) ₂]，7.87[d，J＝8.89Hz，2H，ArH]，8.01[d，J＝8.89Hz，2H，ArH]，8.22[ddd，J＝2.28，4.27，9.19Hz，2H，ArH]，8.38[ddd，J＝2.28，4.27，9.19Hz，2H，ArH]，10.73[s，1H，＝N ⁺(H)-N＝]，11.02[s，1H，ArC(O)NHAr].LCMS(APCI ⁺)：341(100％)。

(E)-N-[4-(1-{[diamino methene base] hydrazono-} ethyl) phenyl]-4-aminobenzamide dihydrochloride (I6).(0.06g 1.05mmol) joins that (0.10g is 0.26mmol) at 2:1 EtOH/H at the acid amides I5 that refluxes with the Fe powder ₂In the suspension among the O (100mL), and the black suspension that obtains refluxed 1.5 hours.After this, Celite pad is passed through in the reaction mixture filtration of heat, and the solvent decompression is removed.Resistates passes through two MeOH-azeotropic cyclic dryings, and then is dissolved in MeOH.This solution with the hcl acidifying in several methyl alcohol, is diluted with EtOAc then.With the suspension filtered that obtains, obtain unbodied brown solid, it is dissolved among the MeOH again.Removal of solvent under reduced pressure, and with resistates be dissolved in again heat H ₂O.The suspension filtered that obtains is passed through Celite pad (using the cold MeOH of a little volume), and the solvent decompression is removed.Resistates obtains amine I6 from the MeOH-EtOAc redeposition, is unbodied yellow solid (0.36g, quantitative yield). ¹H?NMR[CD ₃) ₂SO)：2.25[s，3H，ArC(CH ₃)＝N-]，5.74[s，2H，ArNH ₂]，6.40{v?br?s，4H，＝C(NH ₂) ₂]，6.60[d，J＝8.66Hz，2H，ArH]，7.77[m，6H，ArH]，9.81[s，1H，ArC(O)NHAr]。

(E)-and N-[4-{1-([diamino methene base] hydrazono-) ethyl } phenyl]-4-(6-nitroquinoline-4-base is amino) benzamide hydrochloride salt (Compound I).To amine I6 (～100mg calculates based on I5 ,～0.26mmol) suspension in anhydrous EtOH (60mL) sequentially adds 4-chloro-6-nitroquinoline (220mg 1.04mmol) and dense HCl (3), and refluxes the mixture that obtains 64 hours.After this, reaction mixture is filtered, obtain yellow solid, it from the MeOH-HCl:EtOAc crystallization, is obtained Compound I, be unbodied glassy yellow solid (113mg, 84%). ¹H?NMR[CD ₃) ₂SO)：2.36[s，3H，ArC(CH ₃)＝N-]，7.09[d，J＝6.95Hz，1H?ArH]，7.69[d，J＝8.62Hz，ArH]，7.77[br?s，4H，＝C(NH ₂) ₂]，7.91[d，J＝8.92Hz，2H，ArH]，8.00[d，J＝8.92Hz，2H，ArH]，8.21[d，J＝8.62Hz，2H，ArH]，8.31[d，J＝9.32Hz，2H，ArH]，8.70[d，J＝6.95Hz，1H，ArH]，8.74[dd，J＝9.32，2.29Hz，1H，ArH]，9.86[s，1H，ArH]，10.55[s，1H，ArNHAr]，11.41[s，1H，ArC(O)NHAr]，11.60[brs，1H，1H，＝N ⁺(H)-N＝].LCMS(APCI ⁺)：484(100％)。HPLC：97.1％。

Embodiment J.

4-[4-({ 3-[(1-methyl-4-pyridyl) amino] benzoyl } amino) anilino]-preparation of 6-nitroquinoline dichloride (compound J)

(100mg 0.28mmol) is dissolved in EtOH (10mL) and H with Compound C 6 by heating ₂O (5mL).Add 4-chloro-6-nitroquinoline (F1) (75mg, 0.36mmol) and dense HCl (3), and with reaction mixture refluxed 18 hours (up to using N-BuOH:H ₂The initial amine completely consumed of TLC demonstration of the last phase of the 5:4:1 mixture of O:AcOH is fallen).Reaction mixture dilutes with EtOAc, the backflow several minutes, and make its cooling.The throw out that obtains is filtered and, obtain compound J (148mg, 94%) from the MeOH/EtOAc crystallization; Mp (MeOH, EtOAc)〉300 ℃; ¹H NMR[(CD ₃) ₂SO]: δ 14.90 (br1H, N ⁺H), 11.36 (br, 1H, NH), 10.93 (s, 1H, NH), 10.65 (s, 1H, NH), 9.81 (d, J=2.2Hz, 1H, H-5), 8.71 (dd, J=9.3,2.3Hz, 1H, H-7), 8.60 (d, J=7.0Hz, 1H, H-8), 8.33 (d, J=7.5Hz, 2H, py H-2,6), 8.22 (d, J=9.5Hz, 1H, H-2), 8.02 (d, J=8.9Hz, 2H, ArH), 7.81-7.92 (m, 2H, ArH), 7.68 (t, J=7.8Hz, 1H, ArH), 7.58 (dd, J=8.6,1.3Hz, 1H, ArH), 7.50 (d, J=7.0Hz, 2H, ArH), 7.28 (d, J=7.5Hz, 2H, ArH), 6.91 (d, J=7.0Hz, 1H, ArH), 3.98 (s, 3H, N ⁺CH ₃).

Embodiment K.

4-[3-({ 4-[(6-amino-4-quinolyl base) amino] benzoyl } amino) anilino]-preparation of 1-picoline dichloride (compound K)

(i) Fe powder/EtOH/H ₂O/cat.HCl/ refluxes

(200mg is 0.41mmol) at～6:1 EtOH:H to compound F 17-hydroxy-corticosterone ₂Suspension among the O (6mL) adds Fe powder (200mg), and makes the suspension returning that obtains 3 hours.The reaction mixture of heat is filtered by Celite pad, and with the EtOH washing of Celite pad with heat.The EtOH extracting solution is merged and be evaporated to drying, and resistates is extracted in the water.Solution is filtered by diatomite, be evaporated to drying then.Resistates and methanol azeotropic (3 * 30mL), be dissolved in then among the MeOH (20mL).Add hydrochloric acid in the methyl alcohol (1.25M, 1mL), and with solution stirring 30 minutes.Solution evaporation is arrived drying, and make resistates, obtain compound K (160mg, 80%): mp, (MeOH/EtOAc) 270-274 ℃ from the MeOH/EtOAc crystallization; ¹HNMR[(CD ₃) ₂SO] δ 14.50 (br, 1H, N ⁺H), 10.80 (s, 1H, NH), 10,57 (s, 1H, NH), 10.36 (s, 1H, NH), 8.31 (d, J=7.5Hz, 3H, ArH), 8.15 (d, J=8.6Hz, 2H, ArH), 7.99 (t, J=1.9Hz, 1H, ArH), 7.85 (d, J=9.1Hz, 1H, ArH), 7.68 (br d, J=9.2Hz, 1H, ArH), 7.62 (d, J=8.6Hz, 2H, ArH), and 7.51-7.12 (m, 3H, ArH), 7.23 (d, J=7.6Hz, 2H, ArH), 7.09 (dd, J=7.9,2.8Hz, 1H, ArH), 6.95 (d, J=6.7Hz, 1H, ArH), 3.96 (s, 3H, N ⁺CH ₃), do not observe NH ₂Signal .HRMS (FAB ⁺) calculated value C ₂₈H ₂₅N ₆O (M ^{+ 1}) m/z 461.2090, measured value 461.2108; Ultimate analysis, calculated value C ₂₈H ₂₇N ₆Cl ₃O.0.25H ₂O:C, 58,6; H, 4.8; N, 14.6; Measured value: C, 58.5; H, 4.8, N, 14.5%.

Embodiment L.

4-[4-({ 4-[(6-amino-4-quinolyl base) amino] benzoyl } amino) anilino]-preparation of 1-picoline dichloride (compound L)

(i) Fe powder/EtOH/H ₂O/cat.HCl/ refluxes

To (150mg is 0.28mmol) at 2:1 EtOH:H at the compound G that refluxes ₂Suspension among the O (50mL) adds the Fe powder, and (60mg 1.13mmol), and refluxes the mixture that obtains and spends the night.After this, the reaction mixture of heat is filtered by Celite pad, and solvent reduced pressure once more remove.Resistates is dissolved in MeOH again, and the solvent decompression is removed.This latter's processing repeats twice again, makes resistates from HCl-MeOH:Et then ₂The O recrystallization obtains compound L (33mg, 22%), is unbodied yellow-brown solid. ¹H NMR[(CD ₃) ₂SO]: δ 3.95 (s, 3H, R ₂N ⁺-CH ₃), 6.94 (d, J=6.67Hz, 1H, ArH), 7.13 (d, J=7.36Hz, 2H, ArH), 7.34 (d, J=8.85Hz, 2H, ArH), 7.43 (dd, J=9.06,2.17Hz, 1H, ArH), 7.45 (d, J=1.97Hz, 1H, ArH), 7.62 (d, J=8.61Hz, 2H, ArH), 7.85 (d, J=9.06Hz, 1H, ArH), 7.95 (d, J=8.86Hz, 1H, ArH), 8.15 (d, J=8.59Hz, 2H, ArH), 8.26 (d, J=7.39Hz, 2H, ArH), 8.32 (t, J=6.11Hz, 1H, ArH), 10.35 (s, 1H, ArNHAr), 10.51 (s, 1H, ArNHAr), 10.69 (s, 1H, ArNHAr), 14.49 (brs, 1H, quinolinone-N ⁺-H) [do not observe NH ₂Signal].LCMS(APCI ⁺)：462(100％)。HPLC：97％。

Embodiment M.

6-amino-4-[4-({ 3-[(1-methyl-4-pyridyl) amino] benzoyl } amino) anilino] preparation of quinoline dichloride (compound M)

(i) Fe powder/EtOH/H ₂O/cat.HCl/ refluxes

(80mg is 0.15mmol) at 5:1 EtOH/H to the compound J of vigorous stirring ₂Suspension among the O (5mL) adds Fe powder (43mg), and mixture is refluxed.The dense HCl that adds two then, and continue to reflux 2 hours (up to using N-BuOH:H ₂The TLC demonstration starting raw material completely consumed of the top phase of the 5:4:1 mixture of O:AcOH is fallen).Reaction mixture dilutes with EtOH (100mL) and it is refluxed, with mixture filtration the passing through Celite pad of heat.The top layer resistates of Celite pad is extracted other three times with hot EtOH, to guarantee that amine is extracted fully.The EtOH fraction is merged and be evaporated to drying, and with the resistates hot water extraction.With solution evaporation to dry, and several times with itself and EtOH azeotropic.The resistates that obtains is dissolved among a spot of MeOH, add hydrochloric acid in the methyl alcohol (1.25M, 1mL), and with solution stirring 10 minutes.Then solution is diluted with EtOAc, then some MeOH are evaporated.The throw out that obtains is filtered and, obtain compound M (56mg, 70%) from the MeOH/EtOAc crystallization; Mp〉310 ℃. ¹H?NMR[(CD ₃) ₂SO]δ?14.5(br，1H，N ⁺H)，11.22(s，1H，NH)，10.59(s，1H，NH)，9.55(br，1H，NH)，8.32(d，J＝7.2Hz，2H，ArH)，8.18(d，J＝6.1Hz，1H，ArH)，7.95-7.91(m，4H，ArH)，7.76(d，J＝8.8Hz，1H，ArH)，7.66(t，J＝5.6Hz，1H，ArH)，8.07(d，J＝8.4Hz，1H，ArH)，7.40-7.37(m，3H，ArH)，7.33-7.29(m，3H，ArH)，6.71(d，J＝6.2Hz，1H，ArH)，5.75(brS，2H，NH ₂)，3.98(s，3H，N ⁺CH ₃)。

Embodiment N.

4-[4-({ 4-[(6-amino-4-quinolyl base) amino] anilino } carbonyl) anilino]-preparation of 1-picoline muriate (compound N)

(i) 4-chloro-6-nitroquinoline/EtOH/H ₂O/H+/backflow; (ii) Fe powder/EtOH/H ₂O/cat.HCl/ refluxes

1-methyl-4-[4-(4-[(6-nitro-4-quinolyl) and amino] anilino } carbonyl) anilino] pyridinium chloride (N1).(70mg, 0.33mmol) suspension in MeOH (10mL) adds Compound D 5 (100mg 0.28mmol), and stirs 1 hour (up to obtaining clear soln) with mixture at 20 ℃ to 4-chloro-6-nitroquinoline.Add a dense HCl then, and continue to reflux other 20 hours.TLC analyzes and (uses N-BuOH:H ₂The last phase wash-out of the 5:4:1 mixture of O:AcOH) show some D5 residue, therefore add more 4-chloro-6-nitroquinoline (35mg, 0.16mmol) and continue to reflux other 4 hours.Reaction mixture filters and sequentially uses EtOAc, CH with the EtOAc dilution and with the throw out that obtains then ₂Cl ₂With Di Iso Propyl Ether washing, and, obtain compound N 1 (111mg, 75%), mp (MeOH/EtOAc) at last from the MeOH/EtOAc crystallization〉300 ℃; ¹H NMR[(CD ₃) ₂SO] δ 14.50 (br, 1H, N ⁺H); 11,42, (brs, 1H, NH), 11.10 (s, 1H, NH), 10.58 (s, 1H, NH), 9.82 (d, J=2.1Hz, 1H, ArH), 8.72 (dd, J=9.3,2.2Hz, 1H, ArH), 8.60 (d, J=7.0Hz, 1H, ArH), 8.42 (d, J=7.4Hz, 2H, ArH), 8.24 (d, J=8.6Hz, 1H, ArH), 8.15 (d, J=8.6Hz, 2H, ArH), 8.04 (d, J=8.8Hz, 2H, ArH), 7.51 (t, J=8.3Hz, 4H, ArH), 7.34 (d, J=7.5Hz, 2H, ArH), 6.91 (d, J=7.0Hz, 1H, ArH), 4.01 (s, 3H, N ⁺CH ₃); LCMS 489+ve.

4-[4-(4-[(6-amino-4-quinolyl base) and amino] anilino } carbonyl) anilino]-1-picoline muriate (compound N).(102mg is 0.19mmol) at～5:1 EtOH:H to compound N 1 ₂Solution among the O (3mL) adds Fe powder (53mg) and an AcOH, and with the suspension vigorous stirring that obtains and refluxed 3 hours.The reaction mixture of heat is filtered by diatomite, and Celite pad is washed with more hot EtOH.The EtOH fraction is merged and be evaporated to drying, and resistates is extracted in the hot water.This solution is filtered by diatomite, be evaporated to drying then.Resistates and MeOH azeotropic (3 * 20mL), be dissolved in MeOH (10mL) then.Add hydrochloric acid in the methyl alcohol (1.25M, 2mL), and with solution stirring 10 minutes.Evaporating solvent arrives dry, and with resistates MeOH/EtOAc crystallization, obtains compound N (92mg, 96%) mp (MeOH/EtOAc) ¹H NMR (CD ₃) ₂SO] δ 14.17 (d, J=5.8Hz, 1H NH), 11.11 (s, 1H, NH), 10.52 (s, 1H, NH), 10.21 (s, 1H, NH), 8.37 (d, J=7.5Hz, 2H, ArH), 8.22 (t, J=6.6Hz, 1H, ArH), 8.14 (d, J=8.6Hz, 2H, ArH), 7.98 (d, J=8.9Hz, 2H, ArH), 7.79 (d, J=9.1Hz, 1H, ArH), 7.52-7.59 (m, 3H, ArH), 7.44-7.40 (m, 3H, ArH), 7.43 (d, J=7.6Hz, 2H, ArH), 6.67 (d, J=6.8Hz, 1H, ArH), 4.01 (s, 3H, N ⁺CH ₃) observed part N ⁺H ₃Signal.LCMS?461+ve。

Embodiment 0.

4-[4-({ 4-[(6-dimethylamino-4-quinolyl) amino] benzoyl } amino)-anilino]-preparation of 1-picoline dichloride (compound 0)

To 6-amino-4-quinolyl ketone (01) (112mg, 0.71mmol) and formalin (21.2mmol) solution in EtOH (10mL) sequentially adds NaBH for 40%w/v, 1.6mL ₃CN (335mg, 5.67mmol) and 1N HCl (2.8mL, 2.8mmol), and with the glassy yellow suspension that obtains stirring at room 15 minutes.After this, removal of solvent under reduced pressure is then by two MeOH azeotropic cyclic drying resistatess.With the resistates resuspending in MeOH and filter by diatomite removal of solvent under reduced pressure.Resistates obtains 6-dimethylamino-4-quinolinone (02) (80mg, 60%) from the MeOH:EtOAc crystallization, is unbodied brown solid; ¹HNMR[(CD ₃) ₂SO] δ 2.93[s, 6H, ArN (CH ₃) ₂], 5.90[d, J=7.00Hz, 1H, ArH], 7.13[brs, ArOH], 7.24[m, 2H, ArH] and, 7.45[dd, J=7.41,2.29Hz, 1H, ArH], 7.74[d, J=7.00Hz, 1H, ArH] and .LCMS (APCI ⁺): 189 (100%).R _f＝0.48(10％?MeOH:CH ₂Cl ₂)。

(80mg is 0.43mmol) at POCl with quinolinone 02 ₃Refluxed 1 hour (10mL).After this, excessive POCl is removed in decompression ₃And resistates is dissolved in CH2Cl2, be cooled to 0 ℃, and use ammonia treatment.The mixture that obtains extracts (x3) with CH2Cl2.The organic extraction that merges is sequentially used H ₂MgSO is used in O (x1) and salt solution (x1) washing then ₄Dry.Solvent decompression is removed, obtains 6-dimethylamino-4-chloroquinoline (03) (70mg, 80%), be glassy yellow oily matter people such as [, J.Am.Chem.Soc., 1946,68,1264] Riegel. ¹H NMR[(CD ₃) ₂SO] δ 3.10[s, 6H, ArN (CH ₃) ₂], 6.96[d, J=2.85Hz, 1H, ArH], 7.74[dd, J=9.36,2.85Hz, 1H, ArH], 7.57[d, J=4.69Hz, 1H, ArH] and, 7.90[d, J=9.36Hz, 1H, ArH], 8.46Hz[d, J=4.69Hz, 1H, ArH] and .LCMS (APCI ⁺): 207 (100%), 209 (40%).R _f＝0.73(10％MeOH:CH ₂Cl ₂)。

To being dissolved in 1:2 EtOH:H ₂The amine B7 of O (6mL) (260mg, solution 0.65mmol) sequentially add dense HCl (0.19mL, 6.34mmol) and 03 (140mg, 0.70mmol).The mixture that obtains is refluxed (to be followed the tracks of the reaction progress by TLC, and used N-BuOH:H in 72 hours ₂The last phase wash-out of the 5:4:1 mixture of O:AcOH; Rf=0.30-0.40 shows the glassy yellow spot at 365Nm).After this, the solvent decompression is removed.Remnants are dissolved in MeOH again and solvent reduced pressure once more and remove.The processing of this back repeats once again, and makes resistates from twice crystallization of MeOH:EtOAc, obtains compound 0 (163mg, 45%), is unbodied yellow solid. ¹H?NMR[(CD ₃) ₂SO]δ?3.06(s，6H，Ar(CH ₃) ₂)，3.93(s，3H，R ₂N ⁺-CH ₃)，7.07(d，J＝6.58Hz，2H，ArH)，7.12(d，J＝4.98Hz，1H，ArH)，7.22(d，J＝2.43Hz，1H，ArH)，7.30(d，J＝8.70Hz，2H，ArH)，7.42(m，3H，ArH)，7.78(d，J＝9.28Hz，ArH)，7.92(d，J＝10.75Hz，2H，ArH)，8.01(d，J＝8.64Hz，1H，ArH)，8.19(d，J＝6.93Hz，2H，ArH)，8.32(d，J＝4.90Hz，1H，ArH)，8.50(brs，1H，-NH-)，8.91(s，1H，-NH-)，10.28(s，1H，-NH-)，10.80(v?br?s，1H，-NH-)。LCMS(APCI ⁺)：490(100％)。HPLC：95％。

Embodiment P.

N-[4-(2-amino-6-methylpyrimidine-4-base amino) phenyl]-preparation of 4-(6-(dimethylamino) quinolyl-4 amino) benzamide hydrochloride salt (Compound P)

(i)MeOH，catc.HCl，2.5d

(228mg is 0.62mmol) at 1:2 EtOH:H to acid amides E5 ₂Solution among the O (20mL) sequentially add dense HCl (0.17mL, 5.61mmol) and 4-chloro-6-dimethylamino quinoline (03) (140mg 0.68mmol), and refluxes the mixture that obtains～12 hours.MS afterwards and TLC analyze and (use N-BuOH:H ₂The last phase wash-out of O:AcOH 5:4:1 mixture) be presented at and have some starting raw materials in the reaction mixture, therefore add more 03 (140mg, 0.68mmol).After refluxing several hours, TLC demonstration reaction is finished, thus, and removal of solvent under reduced pressure.Resistates obtains Compound P from the MeOH:EtOAc crystallization, is (very thin) unbodied yellow solid (170mg, 51%). ¹H NMR[(CD ₃) ₂SO] δ 2.28[s, 3H], 3.15[s, 6H, ArN (CH ₃) ₂], 6.21[br s, 1H, ArH], 6.89[d, J=6.71Hz, 1H, ArH], 7.65[m, 5H, ArH] and, 7.82[m, 4H, ArH ﹠amp; ArNH ₂], 7.96[d, J=9.39Hz, 1H, ArH], 8.18[d, J=8.57Hz, 2H, ArH], 8.35[t, J=12.38,6.19Hz, 1H, ArH], 10.44[br s, 1H, ArH], 10.65[br s, 1H, ArNHAr], 10.71[s, 1H, ArNHAr], 12.75[brs, 1H, ArC (O) NH], 14.56[d, J=4.10Hz, 1H, quinolyl N ⁺H] .LCMS (APCI ⁺): 506 (100%).HPLC：97.6％。

Embodiment Q.

6-(dimethylamino)-4-[4-({ 3-[(1-methyl-4-pyridyl) amino] anilino } carbonyl) anilino] preparation of quinoline dichloride (compound Q)

(i) MeOH/cat.

Reflux

(100mg 0.28mmol) is dissolved in EtOH (20mL) and H with amine A7 by heating ₂Among the O (10mL).(70mg 0.33mmol) and dense HCl (3), and refluxes 3 days (up to using N-BuOH:H with mixture to add 4-chloro-6-dimethylamino quinoline (03) then ₂The TLC of the last phase of the 5:4:1 mixture of O:AcOH shows that initial amine is fallen by completely consumed).Reaction mixture is evaporated to drying, and with resistates and EtOH azeotropic.Resistates is dissolved in the MeOH of a little volume, then it is diluted with EtOAc.The throw out that obtains is filtered and, obtain the 163mg crude product, it by preparation property HPLC purifying, is obtained compound Q (70mg, 45%) from the MeOH/EtOAc crystallization; Mp (MeOH/EtOAc),〉300 ℃; ¹H NMR[(CD ₃) ₂SO] δ 14.38 (br, 1H, N ⁺H), 10.71 (s, 1H, NH), 10.56 (s, 1H, NH), 10.52 (s, 1H, NH), 8.36 (d, J=6.7Hz, 1H, ArH), 8.31 (d, J=7.3Hz, 2H, ArH), 8.18 (d, J=8.6Hz, 2H, ArH), 7.99 (brs, 1H, ArH), 7.92 (d, J=9.4Hz, 1H, ArH), 7.69-7.64 (m, 4H, ArH), 7.54 (d, J=2.3Hz, 1H, ArH), 7.49 (t, J=8.1Hz, 1H, ArH), 7.21 (d, J=7.4Hz, 2H, ArH), 7.08 (d, J=7.9Hz, 1H, ArH), 6.91 (d, J=6.7Hz, 1H, ArH), 3.98 (s, 3H, N ⁺CH ₃), 3.14[s, 6H, N (CH ₃) ₂].

Embodiment R.

1-methyl-4-[3-({ 4-[(7-nitro-4-quinolyl) amino] benzoyl } amino) anilino] preparation of pyridinium chloride (compound R)

(i) 4-chloro-7-nitroquinoline/MeOH/cat.HCl/ refluxes

Make A7 and 4-chloro-7-nitroquinoline (people Biorg.Med.Chem.2004 such as RuchelmaN, 12,3731) coupling, obtain compound R with 89% yield; Mp (MeOH/EtOAC), 302-306 ℃ (dec); ¹H NMR[(CD ₃) ₂SO] δ 11.50 (br, 1H, NH), 10.74 (s, 1H, NH), 10.58 (s, 1H, NH), 9.04 (d, J=9.3Hz, 1H, ArH), 8.90 (d, J=2.3Hz, 1H, ArH), 8.77 (d, J=6.7Hz, 1H, ArH), 8.51 (dd, J=9.3,4.5Hz, 1H, ArH), 8.31 (d, J=7.4Hz, 2H, ArH), 8.18 (d, J=8.6Hz, 2H, ArH), 7.98 (t, J=1.9Hz, 1H, ArH), 7.70 (d, J=8.6Hz, 3H, ArH), 7.49 (t, J=8.1Hz, 1H, ArH), 7.22 (d, J=7.6Hz, 2H, ArH), 7.15 (d, J=6.7Hz, 1H, ArH), 7.09 (br d, J=7.8Hz, 1H, ArH), 3.96 (s, 3H, N ⁺CH3); HRMS (FAB ⁺) calculated value C ₂₈H ₂₄N ₆O ₃(M ^{+ 1}) m/z491.1832, measured value 491.1825.

Embodiment S.

1-methyl-4-[4-(4-{7-nitroquinoline-4-base is amino } benzamido)-phenyl amino] preparation of pyridine dichloride (compound S)

(i) 4-chloro-7-nitroquinoline/MeOH/cat.HCl/ refluxes

(1.46g, anhydrous MeOH (20mL) solution 3.73mmol) sequentially add 4-chloro-7-nitroquinoline (S1), and (0.78g 3.73mmol) refluxed 4 hours with two dense HCl and with the mixture that obtains to amine B7.After this, make temperature be reduced to～40 ℃ and continue other 65 hours of heating.After this, removal of solvent under reduced pressure.Resistates is dissolved in MeOH again, and solvent reduced pressure once more removes.The processing of this back repeats once again, makes resistates from MeOH:Et then ₂Twice of O redeposition.At last, make a solid part that so obtains, obtain compound S, be unbodied yellow-orange solids (0.070g, 3%) from the MeOH:EtOAc redeposition. ¹H NMR[(CD ₃) ₂SO]: 3.96[s, 3H, R ₂N ⁺CH ₃], 7.14[m, 3H, ArH], 7.35[d, J=8.85Hz, 2H, ArH], 7.68[d, J=8.57Hz, 2H, ArH], 7.96[d, J=8.85Hz, 2H, ArH], 8.18[d, J=8.57Hz, 2H, ArH], 8.26[d, J=7.37Hz, 2H, ArH], 8.50[dd, J=9.28,2.23Hz, 1H, ArH], 8.77[d, J=6.65Hz, 1H, ArH], 8.94[d, J=2.23Hz, 1H, ArH], 9.08[d, J=9.28Hz, 1H, ArH], 10.55[s, 1H, ArNHAr], 10.71[s, 1H, ArNHAr], 11.21[br s, 1H, ArC (O) NHAr], 11.21[v v br s, 1H, quinoline-N ⁺H]; LCMS (APCI ⁺): 492 (100%), 493 (30%).HPLC：98.1％。

Embodiment T.

4-[4-({ 3-[(1-methyl-4-pyridyl) amino] benzoyl } amino) anilino]-preparation of 7-nitroquinoline dichloride (compound T)

(i) 4-chloro-7-nitroquinoline/MeOH/cat.HCl/reflux.

(92mg 0.25mmol) is dissolved in EtOH (10mL) and H with Compound C 6 by heating ₂Among the O (5mL).(65mg 0.31mmol) refluxes 18 hours (up to using N-BuOH:H with dense HCl (3) and with mixture to add 4-chloro-7-nitroquinoline then ₂The TLC that carries out mutually on the 5:4:1 mixture of O:AcOH shows that initial amine is fallen by completely consumed).Reaction mixture dilutes with EtOAc then, the backflow several minutes, and make its cooling.The throw out that obtains is filtered and, obtain compound T (104mg, 71%) from MeOH/EtOAc crystallization twice; Mp (MeOH, EtOAc)〉300 ℃; ¹H NMR[(CD ₃) ₂SO] δ 14.80 (br1H, N ⁺H), 10.89 (br, 2H, 2 * NH), 10.60 (s, 1H, NH), 8.96 (d, J=9.0Hz, 1H, ArH), 8.84 (bs, 1H, ArH), 8.67 (d, J=6.5Hz, 1H, ArH), 8.32 (d, J=7.3Hz, 2H, ArH), 8.0-7.91 (m, 4H, ArH), 7.68 (t, J=7.8Hz, 1H, ArH), 7.57 (d, J=9.4Hz, 1H, ArH), 7.49 (d, J=8.7Hz, 2H, ArH), 7.29 (d, J=7.4Hz, 2H, ArH), 6.94 (d, J=6.6Hz, 1H, ArH), 3.99 (s, 3H, N ⁺CH ₃).HRMS (FAB ⁺) calculated value C ₂₈H ₂₃N ₆O ₃(M ⁺1) m/z 491.1832, measured value 491.1830.

Embodiment U.

4-[4-({ 4-[(1-methyl-4-pyridyl) amino] benzoyl } amino) anilino]-preparation of 7-nitroquinoline dichloride (compound U)

(i) 4-chloro-7-nitroquinoline/EtOH/H ₂O/H+/backflow

(100mg 0.28mmol) is dissolved in EtOH (20mL) and H with Compound D 5 by heating ₂Among the O (10mL).Add then 4-chloro-7-nitroquinoline (70mg, 0.33mmol) and dense HCl (3) and with reaction mixture refluxed 18 hours (up to using N-BuOH:H ₂The TLC that carries out mutually on the 5:4:1 mixture of O:AcOH shows that initial amine is fallen by completely consumed).Then reaction mixture is diluted with EtOAc, the backflow several minutes, and make its cooling.The throw out that obtains is filtered and, obtain compound U (99mg, 63%) from MeOH/EtOAc crystallization twice; 268 ℃ of mp (MeOH/EtOAc) (dec); ¹H NMR[(CD ₃) ₂SO] δ 10.93 (brs, 2H, 2 * NH), 10.53 (s, 1H, NH), 8.94 (d, J=9.2Hz, 1H, ArH), 8.85 (bs, 1H, ArH), 8.67 (d, J=6.7Hz, 1H, ArH), 8.49 (d, J=9.2Hz, 1H, ArH), 8.37 (d, J=7.5Hz, 2H, ArH), 8.13 (d, J=8.6Hz, 2H, ArH), 8.51 (d, J=8.8Hz, 2H, ArH), 7.50 (t, J=8.7Hz, 4H, ArH), 7.31 (d, J=7.5Hz, 2H, ArH), 6.94 (d, J=6.8Hz, 1H, ArH), 4.02 (s, 3H, N ⁺CH ₃).APCI ⁺ve461。

EXAMPLE V.

4-[3-({ 4-[(7-amino-4-quinolyl base) amino] benzoyl } amino) anilino]-preparation of 1-picoline dichloride (compound V)

(i) Fe powder/EtOH/H ₂O/ refluxes

Carry out the Fe powder reduction of compound R as mentioned above, obtain compound V:mp281-285 ℃ with 83% yield; ¹H NMR[(CD ₃) ₂SO] and δ 13.73 (br, 1H, NH), 10,79 (s, 1H, NH), 10.56 (s, 1H, NH), 10.53 (brs, 1H, NH), 8.41 (d, J=9.3Hz, 1H, ArH), 8.31 (d, J=7.4Hz, 2H, ArH), 8.24 (d, J=7.0Hz, 1H, ArH), 8.14 (d, J=8.6Hz, 2H, ArH), 7.98 (t, J=1.9Hz, 1H, ArH), 7.67 (brd, J=9.2Hz, 1H, ArH), 7.59 (d, J=8.6Hz, 2H, ArH), 7.48 (t, J=8.1Hz, 1H, ArH), 7.23 (d, J=7.5Hz, 2H, ArH), and 7.10-7.06 (m, 2H, ArH), 6.86 (d, J=2.1Hz, 1H, ArH), 6.76 (brs, 2H, NH ₂), 6.68 (d, J=7.0Hz, 1H, ArH), 3.98 (s, 3H, N ⁺CH ₃); HRMS (FAB ⁺) calculated value C ₂₈H ₂₅N ₆O (M ⁺) m/z 461.2090, measured value 461.2108; Ultimate analysis calculated value C ₂₈H ₂₆N ₆ClO ₃.1.25 H ₂O:C, 60.5; H, 5.2; N, 15.1; Cl, 12.8; Measured value: C, 60.6; H, 5.2; N, 15.0; Cl, 13.0%.

Embodiment W.

1-methyl-4-[4-(4-{7-quinolylamine-4-base is amino } benzamido)-phenyl amino] preparation of pyridine dichloride (compound W)

(i) Fe powder, 2:1 EtOH:H ₂O refluxes

To (40mg is 0.07mmol) at 2:1 EtOH:H at the compound S that refluxes ₂Suspension among the O (50mL) adds the Fe powder, and (15mg 0.28mmol), and makes the mixture that obtains reflux several hours up to finishing.After this, the reaction mixture of heat is filtered by Celite pad, and the solvent decompression is removed.Resistates passes through three MeOH cyclic dryings, and at last from the HCl-MeOH:EtOAc recrystallization, obtains compound W (34mg, 90%), is unbodied yellow solid; Mp (MeOH, EtOAc)〉280 ℃; ¹H NMR[(CD ₃) ₂SO]: 3.95 (s, 3H, R ₂N ⁺CH ₃), 6.67 (d, J=7.00Hz, 1H, ArH), 6.92 (d, J=2.13Hz, 1H, ArH), 7.07 (dd, J=9.38,2.13Hz, 1H, ArH), 7.18 (br d, J=6.45Hz, 2H, ArNH ₂), 7.34 (d, J=8.89Hz, 2H, ArH), 7.60 (d, J=8.60Hz, 2H, ArH), 7.97 (d, J=8.89Hz, ArH), 8.16 (d, J=8.60Hz, 2H ArH), 8.22 (t, J=6.66Hz, 1H, ArH), 8.27 (d, J=7.49Hz, 2H, ArH), 8.51 (d, J=9.39Hz, 1H, ArH), 10.58 (s, 1H, ArNHAr), 10.71 (s, 1H, ArNHAr), 11.00 (s, 1H, ArC (O) NHAr), 14.08 (d, J=6.09Hz, 1H, quinoline-N ⁺-H); LCMS (APCI ⁺): 462 (100%); HPLC:96.1%.

Embodiment X.

7-amino-4-[4-({ 3-[(1-methyl-4-pyridyl) amino] benzoyl } amino) anilino] preparation of quinoline dichloride (compounds X)

(i) Fe powder/EtOH/H ₂O/ refluxes

(107mg is 0.19mmol) at 5:1 EtOH:H to the compound T of vigorous stirring ₂Suspension among the O (5mL) adds Fe powder (43mg) night, and mixture is refluxed.Add two dense HCl, and continue to reflux 2 hours (up to using N-BuOH:H ₂The TLC demonstration starting raw material completely consumed of carrying out mutually on the 5:4:1 mixture of O:AcOH is fallen).Reaction mixture is used EtOH (100mL) dilution then, and makes its backflow.The mixture of heat is filtered by Celite pad, and the top layer of Celite pad is extracted three times to guarantee that amine is extracted fully with hot EtOH.The ethanol extract that merges is evaporated to drying, and with the hot H of resistates ₂O extracts.With solution evaporation to dry, and with the EtOH azeotropic several times.The resistates that obtains is dissolved among the MeOH of a little volume, add hydrochloric acid in the methyl alcohol (1.25M, 1mL), and with solution stirring 10 minutes.Solution dilutes with EtOAc, and with some MeOH evaporations.The throw out that obtains is filtered and, obtain compounds X (99mg, 98%) from the MeOH/EtOAc crystallization; Mp (MeOH/EtOAc〉300 ℃; ¹H NMR[(CD ₃) ₂SO] δ 13.48 (s, 1H, N ⁺H), 10.98 (s, 1H, NH), 10.59 (s, 1H, NH), 10.32 (s, 1H, NH), 8.36-8.31 (m, 3H, ArH), 8.14 (d, J=7.0Hz, 1H, ArH), 8.00-7.91 (m, 4H, ArH), 7.67 (t, J=7.7Hz, 1H, ArH), 7.57 (d, J=7.4H, 1H, ArH), 7.40 (d, J=8.6Hz, 2H, ArH), 7.02 (d, J=9.2Hz, 1H, ArH), 6.82 (bs, 1H, ArH), 6.66 (brs, 2H, NH ₂), 6.43 (d, J=6.9Hz, 1H, ArH), 3.99 (s, 3H, N ⁺CH ₃).HRMS (FAB ⁺) calculated value C ₂₈H ₂₅N ₆O (M ^{+ 1}) m/z 461.2090, measured value 461.2085.

Embodiment Y.

4-[4-({ 4-[(7-amino-4-quinolyl base) amino] anilino } carbonyl) anilino]-preparation of 1-picoline muriate (compound Y)

(i) Fe powder/EtOH/H ₂O/ refluxes

(101mg is 0.19mmol) at～5:1 EtOH:H to compound U ₂Suspension among the O (3mL) adds Fe powder (53mg), adds an AcOH subsequently.With the suspension vigorous stirring that obtains and refluxed 3 hours.The reaction mixture of heat is filtered by diatomite, and with the EtOH washing of Celite pad with heat.The EtOH extracting solution that merges is evaporated to drying and resistates is extracted in the hot water.Solution is filtered by diatomite, be evaporated to drying then.Resistates and MeOH azeotropic (3 * 20mL), be dissolved in MeOH (10mL) then.Add hydrochloric acid in the methyl alcohol (1.25M, 2mL), and with solution stirring 10 minutes.Then with solution evaporation to dry, and with resistates from the MeOH/EtOAc crystallization, obtain 290-295 ℃ of compound Y (55mg, 58%) mp (MeOH/EtOAc); ¹H NMR (CD ₃) ₂SO] δ 13.44 (br, 1H, N ⁺H), 11.04 (s, 1H, NH), 10.50 (s, 1H, NH), 10.31 (s, 1H, NH), 8.38-8.33 (m, 3H, ArH), 8.15-8.11 (m, 3H, ArH), 7.97 (d, J=8.9Hz, 2H, ArH), 7.51 (d, J=8.7Hz, 2H, ArH), 7.41 (d, J=9.9Hz, 2H, ArH), 7.33 (d, J=7.5Hz, 2H, ArH), 7.03 (dd, J=9.2,2.1Hz, 1H, ArH), 6.81 (d, J=2.2Hz, 1H, ArH), 6.67 (bs, 2H, NH ₂), 6.44 (d, J=7.1Hz, 1H, ArH), 4.01 (s, 3H, N ⁺CH ₃); LCMS 461+ve.

Embodiment Z.

7-(dimethylamino)-4-[4-({ 3-[(1-methyl-4-pyridyl) amino] anilino } carbonyl) anilino] preparation of quinoline dichloride (compound Z)

(i) EtOH/H ₂O/cat.

Reflux

(70mg, (100mg in EtOH 0.28mmol) (20ml) solution, and adds H 0.34mmol) to join A7 with 4-chloro-7-dimethylamino quinoline (Z1) people such as [, WO 9611187A1] KoNishi ₂O (10mL), (0.25mL 9eq), and refluxed 2 days to add dense HCl then.The mass spectrum of the sample of reaction mixture shows that A7 still exists, therefore add more 4-chloro-7-dimethylamino quinoline (35mg, 0.17mmol) and continue to reflux other 5 days.Then reaction mixture is evaporated to drying.Resistates is dissolved in the MeOH of a little volume, dilutes with EtOAc.The throw out that obtains is filtered, and from the MeOH/EtOAc crystallization, obtain crude product (160mg), (with total mass number is 489 by preparation property HPLC purifying with it ⁺The fraction of ve is evaporated to drying, and resistates is dissolved in EtOH/H ₂O 2:1 (5mL) with the EtOAc dilution, and with the throw out filtration that obtains, obtains compound Z (20mg 13%).Analyzing by HPLC is 90% purity; MP (MeOH EtOAc) 285-289 ℃ (dec); ¹HNMR[(CD3) 2SO] δ 13.47 (br, 1H, N ⁺H) 10,54 (s, 1H, NH), 10.51 (s, 1H, NH), 10.49 (s, 1H, NH), 8.44 (d, J=9.7Hz, 1H, ArH), 8.35-8.30 (m, 3H, ArH) 8.13 (d, J=8.6Hz, 2H, ArH), 7.98 (t, J=1.9Hz, 1H, ArH), 7.63-7.58 (m, 3H, ArH), 7.49 (t, J=8.1Hz, 1H, ArH), 7.40 (dd, J=9.6,2.5Hz, 1H, ArH), 7.18 (d, J=7.5Hz, 2H, ArH), 7.09 (dd, J=7.7,1.3Hz, 1H, ArH), 6.81 (d, J=2.5Hz, 1H, ArH), 6.71 (d, J=7.0Hz, 1H, ArH), 3.98 (s, 3H, N ⁺CH ₃), 3.16 (s, 6H, NMe ₂); Mass spectrum APCI ⁺Ve 489.

Embodiment A A.

4-[4-({ 4-[(7-dimethylamino) quinolyl-4 amino) benzamido)-phenyl amino)-preparation of 1-picoline dichloride (compd A A)

(i) EtOH/H ₂O/cat.

Reflux

(311mg 0.79mmol) is dissolved in 1:2 EtOH:H to amine B7 ₂The solution of O (3mL) sequentially add dense HCl (0.22mL, 0.72mmol) and 4-chloro-7-dimethylamino quinoline (Z1) (175mg, 0.85mmol).The mixture backflow that obtains (was followed the tracks of reaction process by TLC, and used N-BuOH:H in 30 hours ₂The last phase wash-out of O:AcOH 5:4:1 mixture; Rf=0.30-0.40 is the glassy yellow spot at 365Nm).After this, removal of solvent under reduced pressure.Resistates is dissolved in MeOH again, and solvent reduced pressure once more removes.The processing of this back repeats once again, and resistates obtains compd A A from MeOH:EtOAc crystallization twice, is unbodied yellow solid (93mg, 21%). ¹H?NMR[CD ₃) ₂SO]：3.15[s，6H，ArN(CH ₃) ₂]，3.96[s，3H，ArN ⁺-CH ₃]，6.72[d，J＝7.00Hz，1H，ArH]，6.91[d，J＝2.36Hz，1H?ArH]，7.14[d，J＝7.40Hz，2H，ArH]，7.36[m，3H，ArH]，7.62[d，J＝8.53Hz，2H，ArH]，7.95[d，J＝8.89Hz，2H，ArH]，8.16[d，J＝8.53Hz，2H，ArH]，8.26[d，J＝7.40Hz，2H，ArH]，8.32[d，J＝7.00Hz，1H，ArH]，8.57[d，J＝9.59Hz，1H，ArH]，10.53[s，1H，-NH-]，10.70[s，1H，-NH-]，10.75[s，1H，-NH-]，13.85[brs，1H，Ar-N ⁺H].LCMS(APCI ⁺)：490(100％)，491(20％)。HPLC：96.7％。

Embodiment B B.

N-[4-(2-amino-6-methylpyrimidine-4-base amino) phenyl]-preparation of 4-(7-(dimethylamino) quinolyl-4 amino) benzamide hydrochloride salt (compd B B)

To acid amides E5 (306mg, 1:2 EtOH:H 0.83mmol) ₂O (10mL) solution sequentially adds dense HCl (0.23mL) and 4-chloro-7-dimethylamino quinoline (Z1), and (188mg 0.91mmol), and refluxes the mixture that obtains 20 hours.[MS and TLC at reaction mixture analyze (5:4:1 N-BuOH:H afterwards ₂O:CH ₃CO ₂H) show when remaining a small amount of E5 is degrading], reaction mixture is filtered, and the solid that obtains is sequentially used EtOAc and hexane wash, obtain compd B B, be unbodied lemon-yellow solid (199mg, 45%). ¹HNMR[CD ₃) ₂SO]: 2.28[s, 3H, ArCH ₃], 3.15[s, 6H, ArN (CH ₃) ₂], 6.18[br s, 1H, ArH], 6.70[d, J=7.02Hz, 1H, ArH], 6.88[d, J=2.46Hz, 1H, ArH] and, 7.38[dd, J=9.62,2.46Hz, 1H, ArH], 7.61[d, J=8.60,2H, ArH] and, 7.81[m, 5H, ArH ﹠amp; ArNH ₂], 8.14[d, J=8.60Hz, 2H, ArH], 8.31[d, J=7.02Hz, 1H, ArH], 8.55[d, J=9.71Hz, 1H, ArH] and, 10.41[brs, 1H, ArH], 10.65[m, 2H, ArNHAr ﹠amp; ArNHAr], 12.68[v br s, 1H, ArC (O) NH], 13.77[vbr s, 1H, quinolyl N ⁺H] .LCMS (APCI ⁺): 506 (100%).HPLC：96.6％。

Embodiment C C.

7-(dimethylamino)-4-[4-({ 3-[(1-methyl-4-pyridyl) amino] benzoyl } amino) aniline] preparation of quinoline dichloride (Compound C C)

(i) 4-chloro-7-(dimethylamino) quinoline/EtOH/H ₂O/H ⁺/ reflux.

(71mg, 0.34mmol) (0.3mL, (104mg is 0.0.29mmol) at EtOH (10ml) and H 9eq) sequentially to join C6 with dense HCl with 4-chloro-7-dimethylamino quinoline (Z1) ₂In the solution among the O (5mL), and with mixture backflow 4 days.Reaction mixture refluxes, then cool to room temperature with EtOAc (150mL) dilution then.The throw out that obtains is filtered, be dissolved in the MeOH of a little volume then.Solution dilutes with EtOAc, and the throw out that obtains is filtered, and by preparation property HPLC purifying, obtains Compound C C (13mg, 8%): 188 ℃ of mp (MeOH/EtOAc) (dec) then; ¹HNMR[(CD ₃) ₂SO] δ 13.22 (bd, J=5.7Hz, 1H, N ⁺H), 10.61 (s, 1H, NH), 10.51 (s, 1H, NH), 10.38 (s, 1H, NH), 8.42 (d, J=9.7Hz, 1H, ArH), 8.32 (d, J=7.5Hz, 2H, ArH), 8.23 (t, J=6.7Hz, 1H, ArH), 7.96-7.92 (m, 3H, ArH), 7.88 (t, J=1.8Hz, 1H, ArH), 7.69 (t, J=7.9Hz, 1H, ArH), 7.58 (dd, J=8.0,1.4Hz, 1H, ArH), 7.43 (d, J=8.9Hz, 2H, ArH), 7.34 (dd, J=9.6,2.6Hz, 1H, ArH), 7.21 (d, J=7.5Hz, 2H, ArH), 6.77 (d, J=2.5Hz, 1H, ArH), 6.48 (d, J=7.1Hz, 1H, ArH), 3.99 (s, 3H, N ⁺CH ₃), 3.15[s, 6H, N (CH ₃) ₂]. mass spectrum APCI ⁺Ve 489.

Embodiment DD.

7-(dimethylamino)-4-[4-({ 4-[(1-methyl-4-pyridyl) amino] benzoyl } amino) aniline] preparation of quinoline dichloride (Compound D D)

(i) EtOH/H ₂O/cat.

Reflux

(140mg, 0.62mmol) (0.3mL, (110mg is 0.31mmol) at EtOH (20ml) and H 9eq) sequentially to join D5 with dense HCl with 4-chloro-7-dimethylamino quinoline (Z1) ₂In the solution among the O (10mL), and mixture refluxed the 4d. reaction mixture with EtOAc (150mL) dilution, reflux, then cool to room temperature.The throw out that obtains is filtered, be dissolved in the MeOH of a little volume then, then it is diluted with EtOAc.The throw out that obtains is filtered, obtain crude product, it by preparation property HPLC purifying, is obtained Compound D D (47mg, 27%) HPLC 99.9%; 198-200 ℃ of mp (MeOH/EtOAc); ¹HNMR[(CD ₃) ₂SO] δ 13.26 (br, 1H, N ⁺H), 10.71 (s, 1H, NH), 10.46 (s, 1H, NH), 10.37 (brs, 1H, NH), 8.43 (d, J=9.6Hz, 1H, ArH), 8.37 (d, J=7.4Hz, 2H, ArH), 8.23 (d, J=7.0Hz, 1H, ArH), 8.11 (d, J=8.6Hz, 2H, ArH), 7.96 (d, J=8.8Hz, 2H, ArH), 7.50 (d, J=8.6Hz, 2H, ArH), 7.43 (d, J=8.8Hz, 2H, ArH), 7.34 (dd, J=9.6,2.4Hz, 1H, ArH), 7.28 (d, J=7.5Hz, 2H, ArH), 6.78 (d, J=2.4Hz, 1H, ArH), 6.49 (d, J=7.0Hz, 1H, ArH), 4.01 (s, 3H, N ⁺CH ₃), 3.14[s, 6H, N (CH ₃) ₂]. mass spectrum APCI ⁺Ve 489.

Table 2.

Exemplary compounds of the present invention

For the clauses and subclauses in the table 2, R ⁴Be H, A is NH, and is H with X.Except as otherwise noted, R ⁶-R ⁹Be H.Except as otherwise noted, G7, G8, D1, and be CH with D2.

Compound	R 6-R 9	Y	Connect	Z	mp	Molecular formula	HPLC(％)	Analyze
									A	--	CONH	m	Q1	253-257	C 28H 27N 5OCl 2	99.7	C，H，N
V	R 8＝NH 2	CONH	m	Q1	281-285	C 28H 26N 6OCl 2	99.9	C，H，N
									K	R 7＝NH 2	CONH	m	Q1	270-274	C 28H 26N 6OCl 2	97.9	C，H，N
L	R 7＝NH 2	CONH	p	Q1	262-266	C 28H 26N 6OCl 2	99.3	LRMS
									0	R 7＝NMe 2	CONH	p	Q1	242-246	C 30H 30N 6OCl 2	96.4	LRMS
N	R 7＝NH 2	NHCO	p	Q1	>300	C 28H 26N 6OCl 2	97.6	LRMS
									F	R 7＝NO 2	CONH	m	Q1	273-277	C 28H 24Cl 2N 6O 3	97.0	HRMS
R	R 8＝NO 2	CONH	m	Q1	303(dec.)	C 28H 24Cl 2N 6O 3	96.0	HRMS
									Y	R 8＝NH 2	NHCO	p	Q1	290-295	C 28H 26N 6Ocl 2	97.0	HRMS
S	R 8＝NO 2	CONH	p	Q1	265(dec.)	C 28H 24Cl 2N 6O 3	98.1	LRMS
									AA	R 8＝NMe 2	CONH	p	Q1	242-245	C 30H 30Cl 2N 6O	97.6	LRMS
J	R 7＝NO 2	NHCO	m	Q1	>300	C 28H 24N 6Cl 2O 3	98.5	HRMS
									T	R 8＝NO 2	NHCO	m	Q1	>300	C 28H 24N 6Cl 2O 3	97.7	HRMS
D	--	NHCO	p	Q1	>300	C 28H 25NCl 2N 5O	96.6	HRMS
									C	--	NHCO	m	Q1	>300	C 28H 25NCl 2N 5O	97.5	HRMS
U	R 8＝NO 2	NHCO	p	Q1	268dec	C 28H 24N 6Cl 2O 3	96.6	LRMS
									M	R 7＝NH 2	NHCO	m	Q1	>310	C 28H 26N 6OCl 2	96.0	HRMS
X	R 8＝NH 2	NHCO	m	Q1	>300	C 28H 26N 6OCl 2	95.8	HRMS
									BB	R 8＝NMe 2	CONH	p	Q2	272-275	C 29H 29ClN 8O	96.6	HRMS
EEE7	R 7＝NMe 2	CONH	p	Q2	260-265	C2 9H 29ClN 8O	97.6	LRMS
									H	R 7＝NO 2	CONH	p	Q2	301-305	C 27H 23ClN 8O 3	98.5	LRMS
Q	R 7＝NMe 2	CONH	m	Q1	>300	C 30H 30Cl 2N 6O	97.8	LRMS
									Z	R 8＝NMe 2	CONH	m	Q1	285-289	C 30H 30Cl 2N 6O	90.0	HRMS

Compound	R 6-R 9	Y	Connect	Z	mp	Molecular formula	HPLC(％)	Analyze
									B	--	CONH	p	Q1	>300	C 28H 25Cl 2N 5O	99.7	LRMS
DD	R 8＝NMe 2	NHCO	p	Q1	198-200	C 30H 30Cl 2N 6O	99.9	HRMS
									CC	R 8＝NMe 2	NHCO	m	Q1	188dec	C 30H 30Cl 2N 6O	98.5	HRMS
E	--	CONH	p	Q2	189-192	C 27H 24ClN 7O	95.7	LRMS
									I	R 7＝NO 2	CONH	p	Q3	>300	C 25H 23ClN 8O	97.1	LRMS
G	R 7＝NO 2	CONH	p	Q1	183-187	C 28H 24Cl 2N 6O 3	99.9	LRMS
									EE	R 7＝NMe 2	NHCO	m	Q2	>300	C 29H 29ClN 8O	96.0	HRMS
W	R 8＝NH 2	CONH	p	Q1	242-245	C 28H 26Cl 2N 6O	96.1	LRMS
									002	R 7＝NMe 2	NHCO	p	Q1	156	C 30H 30Cl 2N 6O	97.0	HRMS
NN	R 7＝NMe 2	CONH	p	Q3	>280	C 27H 30Cl 2N 8O	98.7	LRMS
									001	R 7＝NMe 2	NHCO	m	Q1	170-173	C 30H 30Cl 2N 6O	98.0	HRMS
QQ	R 7＝NMe 2	NHCO	m	Q3	>280	C 27H 30Cl 2N 8O	96.0	HRMS
									EEE2	--	CONH	p	Q9	171-174	C 27H 23Cl 2N 5O	100.0	LRMS
PP	R 7＝NMe 2	NHCO	p	Q3	>300	C 27H 30Cl 2N 8O	97.9	HRMS
									GG1	--	NHCO	p	Q2	>300	C 27H 25Cl 2N 7O	97.7	HRMS
GG2	R 8＝NH 2	NHCO	p	Q2	257-261	C 27H 26Cl 2N 8O	97.8	HRMS
									JJ1	R 7＝NMe 2	CONH	m	Q3	278-282	C 27H 30Cl 2N 8O	95.8	LRMS
JJ2	--	NHCO	m	Q3	264-268	C 25H 25Cl 2N 7O	96.4	LRMS
									GG3	R 7＝NH 2	NHCO	p	Q2	>300	C 27H 26Cl 2N 8O	96.4	HRMS
II	--	CONH	m	Q2	251-255	C 27H 25Cl 2N 7O	95.4	LRMS
									FF1	--	NHCO	m	Q2	>300	C 27H 25Cl 2N 7O	99.5	HRMS
FF2	--	NHCO	m	Q4	>300	C 26H 24Cl 2N 8O	99.2	HRMS
									LL	R 7＝Cl	CONH	p	Q2	250-254	C 27H 24Cl 3N 7O	99.7	LRMS
KK	R 7，R 8，R 9＝F	CONH	?p	Q2	>280	C 27H 22Cl 2F 3N 7O	98.9	LRMS
									HH	R 7＝NMe 2	NHCO	p	Q2	>300	C 29H 30Cl 2N 8O	96.7	HRMS
GG4	R 7＝NO 2	NHCO	p	Q2	295-300	C 27H 24Cl 2N 8O 3	96.4	HRMS
									GG5	R 8＝NO 2	NHCO	p	Q2	>300	C 27H 24Cl 2N 8O 3	96.6	HRMS
RR1	R 7＝NO 2	NHCO	m	Q4	>300	C 26H 23Cl 2N 9O 3	96.0	HRMS
									RR2	R 7＝NO 2	NHCO	m	Q2	>300	C 27H 24Cl 2N 8O 3	99.0	HRMS
RR3	R 7＝NH 2	NHCO	m	Q2	280-284	C 27H 26Cl 2N 8O	98.0	LRMS
									N1	R 7＝NO 2	NHCO	p	Q1	>300	C 28H 24Cl 2N 6O 3	97.0	LRMS
SS	R 7＝NMe 2	NHCO	p	Q4	283-287	C 28H 29Cl 2N 9O	99.2	RMS
									TT	--	NHCO	p	Q4	263-267	C 26H 24Cl 2N 8O	98.3	LRMS

Compound	R 6-R 9	Y	Connect	Z	mp	Molecular formula	HPLC(％)	Analyze
									UU	R 7＝NH 2	NHCO	m	Q4	250-255	C 26H 25Cl 2N 9O	99.5	HRMS
VV1	R 7＝NO 2	NHCO	p	Q4	260-265	C 26H 23Cl 2N 9O 3	100.0	HRMS
									VV2	R 7＝NH 2	NHCO	p	Q4	262-266	C 26H 25Cl 2N 9O	99.7	HRMS
WW	--	CONMe	p	Q9	287-292	C 28H 25Cl 2N 5O	96.3	HRMS
									XX*	--	NHCO	p	Q2	>290	C 26H 24Cl 2N 8O	99.2	C，H，N
YY**	--	NHCO	p	Q2	>290	C 26H 24Cl 2N 8O	98.9	C，H，N
									ZZ***	--	NHCO	p	Q2	>295	C 26H 24Cl 2N 8O	100.0	C，H，N

*G ₈＝N；**D ₂＝N；***D ₁＝N

Embodiment E E.

4-[4-({ 3-[(2-amino-6-methyl-4-pyrimidyl) amino] benzoyl } amino) anilino]-6-(dimethylamino) quinoline muriate (compd E E) synthetic

(i) the anhydrous HCl/ of EtOH/ refluxes; (ii) H ₂/ 10%Pd/C/MeOH; (iii) NaBH ₃CN/HCHO/NaOAc/MeOH/HCl/RT; (iv) .2NHCl/EtOH refluxes; (v) cellosolvo/HCl/ refluxes; (vi) compd A 6/EDCI/DMAP/DMF/RT.

4-[4-(acetylamino) anilino]-6-nitroquinoline muriate (A3).To N-(4-aminophenyl) ethanamide (A1) (933mg, 6.2mmol) ethanol (30mL) solution add and to contain 6-nitro-4-chloroquinoline (A2) (1.08g, 5.18mmol) ethanol (10mL), add subsequently and contain 1 of 4N HCl, 4-dioxane solution (1.5mL).Made reaction mixture refluxed 30 minutes, at this moment, TLC (SiO ₂/ 8%MeOH/DCM) the demonstration reaction is finished.Reaction mixture makes its boiling with the EtOAc dilution, it is cooled to 20 ℃ then.The throw out that obtains is filtered,, obtain A3 (1.79g, 96%) with the EtOAc washing and from the MeOH recrystallization; Mp (MeOH)〉280 ℃; ¹H NMR[(CD ₃) ₂SO] δ 14.59 (br, 1H, N ⁺H), 11.36 (br, 1H, NH), 10.24 (s, 1H, NH), 9.78 (d, J=2.3Hz, 1H, ArH), 8.71 (dd, J=9.3,2.3Hz, 1H, ArH), 8.57 (d, J=7.0Hz, 1H, ArH), 8.21 (d, J=9.3Hz, 1H, ArH), 7.80 (d, J=8.8Hz, 2H, ArH), 7.42 (d, J=8.8Hz, 2H, ArH), 6.86 (d, J=7.0Hz, 1H, ArH), 2.09 (s, 3H, COCH ₃), APCI ⁺Ve 323.

4-[4-(acetylamino) anilino]-6-quinolylamine muriate (A4).To ((1.75g, 4.88mmol) adding 10%Pd/C (0.1g) of the suspension in MeOH (10mL) and hydrogenation (under 40psi pressure) are 2 hours for A3.Reaction mixture is filtered and wash with more MeOH.Filtrate is concentrated to half volume, evaporates with the EtOAc dilution and with remaining MeOH.The throw out that obtains filters, and with more EtOAc washing and dry, obtains pure basically (9) (1.255g78%); Mp (MeOH/EtOAc)〉280 ℃; ¹H NMR[(CD ₃) ₂SO] δ 13.95 (br, 1H, N ⁺H), 10.14 (s, 1H, NH), 10.01 (s, 1H, NH), 8.10 (d, J=6.7Hz, 1H, ArH), 7.74 (d, J=8.9Hz, 3H, ArH), 7.43 (d, J=2.2Hz, 1H, ArH), 7.38-7.32 (m, 3H, ArH), 6.61 (d, J=6.7Hz, 1H, ArH), 5.94 (s, 2H, NH ₂), 2.08 (s, 3H, COCH ₃); APCI+ve 293.

N-(4-{[6-(dimethylamino)-4-quinolyl] amino } phenyl) ethanamide (A5).To A4 (1.237g, MeOH 3.76mmol) (50mL) solution add 38% formalin (8.4mL, 112.8mmol, 30eq), NaBH3CN (2.134g, 30.08mmol, 8eq), NaOAc (526mg, 6.41mmol) and 2 dense HCl.Reaction mixture was stirred 2 hours at 20 ℃, at this moment, TLC (SiO ₂/ 8%MeOH/DCM/aq NH3) and mass spectrum show that reaction finishes.Then with reaction mixture with dense HCl acidifying, stirred 1 hour at 20 ℃.Afterwards, reaction mixture is used NH carefully ₃Aqueous solution neutralization.With the MeOH reduction vaporization and with aqueous resistates and NH ₃The aqueous solution stirs together.The throw out that obtains is filtered, wash with water, obtain pure basically A5 (1.22g, 91%).With it without being further purified use; Mp, (MeOH)〉280 ℃; ¹H NMR[(CD ₃) ₂SO] δ 9.95 (s, 1H, NH), 8.85 (bs, 1H, NH), 8.18 (d, J=5,5Hz, 1H, ArH), 7.72 (d, J=9.3Hz, 1H, ArH), 7.65 (d, J=8.8Hz, 2H, ArH), 7.41 (dd, J=9.3,2.6Hz, 1H, ArH), 7.29-7.26 (m, 3H, ArH), 6.67 (d, J=5.5Hz, 1H, ArH), 3.06[s, 6H, N (CH ₃) ₂], 2.06 (s, 3H, COCH ₃); APCI ⁺Ve 321.

4-(4-amino-benzene amido)-6-(dimethylamino) quinoline muriate (A6).1, the suspension in the 4-diox (20mL) adds 1.5Maq HCl (5mL) and made reaction mixture refluxed 2 hours to A5.Then reaction mixture is diluted with MeOH/EtOAc, make its backflow and it is cooled to 20 ℃.The throw out that obtains is filtered,, obtain pure basically A6, be light yellow solid (1.20g, 100%) with EtOAc washing and dry; Mp (MeOH/EtOAc) 152-155; ¹H NMR[(CD ₃) ₂SO] 14.35 (br, 1H, N ⁺H), 10.50 (br, 2H, NH ₂), 8.27 (d, J=6.6Hz, 1H, ArH), 7.92 (d, J=9.4Hz, 1H, ArH), 7.65 (dd, J=9.5,2.6Hz, 1H, ArH), 7.57 (d, J=2.4Hz, 1H, ArH), 7.54 (d, J=8.5Hz, 2H, ArH), 7.41 (d, J=8.1Hz, 2H, ArH), 6.68 (d, J=6.7Hz, 1H, ArH), 3.12 (s, 6H, [N (CH ₃) ₂]; APCI ⁺Ve 279.

3-[(2-amino-6-methyl-4-pyrimidyl) amino] benzoate hydrochlorate (A9).By heating make 3-benzaminic acid (A7) (1.02g, 7.42mmol) and 2-amino-6-chloro-4-methylpyrimidine (A8) (1.08g 7.42mmol) is dissolved in cellosolvo, adds a dense HCl then, refluxes 1 hour and is cooled to 20 ℃.The throw out that obtains is filtered,, wash with EtOAc subsequently with more cellosolvo washing.Solid obtains A9 (1.98g, 95%) from the MeOH/EtOAc recrystallization, is pale solid; Mp〉300 ℃; ¹H NMR[(CD ₃) ₂SO] δ 12.97 (bs 2H, NH, and with COOH), 10.81 (s, 1H, NH), 8.22 (bs, 1H, ArH), 8.03 (s, 1H, ArH), 7.82 (br, 2H, NH ₂), 7.72 (bd, J=7.7Hz, 1H, ArH), 7.50 (t, J=7.9Hz, 1H, ArH), 6.22 (s, 1H, ArH); APCI ⁺Ve 245.

4-[4-(3-[(2-amino-6-methyl-4-pyrimidyl) and amino] benzoyl } amino) anilino]-6-(dimethylamino) quinoline muriate (15) (compd E E).With 4-(4-amino-benzene amido)-6-(dimethylamino) quinoline muriate (A6) (102mg, 0.32mmol), A9 (111mg, 0.32mmol), and with EDCI (151mg, 0.64mmol) mixture in DMF (5mL) stirred 5 minutes at 20 ℃.(96mg 0.64mmol) and with reaction mixture stirred 72 hours at 20 ℃ to add DMAP then.Solvent decompression removed and with resistates H ₂The O dilution is also used NH ₃Aqueous solution alkalization.The throw out that obtains is filtered, wash with water, and pass through at SiO ₂In chromatography carry out purifying, with 0-5% DCM/MeOH and 1% NH ₃The gradient elution of the aqueous solution obtains 102mg 62%.(98mg 0.19mmol) is dissolved in MeOH (5mL) and add and to contain 1 of 4N HCl, 4-diox (0.3mL) and stirring 30 minutes with its sample.Then it is filtered with EtOAc dilution and with the throw out that obtains and, obtain compd E E (107mg) from the MeOH/EtOAc recrystallization; ¹H NMR[(CD ₃) ₂SO] δ (13.9br1H, N ⁺H), 12.9 (br (1H, N ⁺H), 10.58 (s, 2H, 2 * NH), 10.40 (s, 1H, NH), 8.26 (d, J=6.8Hz, 1H, ArH), 8.38 (br1H, ArH), 8.06 (bs, 1H, ArH), 8.00 (d, J=8.9Hz, 2H, ArH), 7.89 (d, J=9.4Hz, 1H, ArH), 7.75 (bd, J=7.6Hz, 1H, ArH), 7.56-7.52 (m, 2H, ArH), 7.46 (d, J=8.8Hz, 2H, ArH), 6.67 (d, J=6.8Hz, 1H), 6.19 (s, 1H, ArH), 3.12[s, 6H, (NCH ₃) ₂], 2.28 (s, 3H, CH ₃), do not observe NH ₂Signal, APCI ⁺Ve 505.

Embodiment F F.

3-[(2-amino-6-methyl-4-pyrimidyl) amino]-N-[4-(4-quinolyl amino) phenyl] preparation of benzamide dihydrochloride (compound F 17-hydroxy-corticosterone F1) and relevant compound (compound F 17-hydroxy-corticosterone F2)

Reagent: (i) cellosolvo/H ⁺/ reflux; (ii) H ₂/ 10%Pd/C/MeOH; (iii) 4-chloroquinoline/EtOH/H ₂O/ refluxes

3-[(2-amino-6-methyl-4-pyrimidyl) amino]-N-(4-nitrophenyl) benzamide hydrochloride salt (B2).(1.0g, 3.89mmol) (569mg 3.89mmol) is dissolved in warm cellosolvo (20mL) with 4-chloro-6-methyl-2-pyrimidyl amine (A8) with 3-amino-N-(4-nitrophenyl) benzamide (B1).Add two dense HCl and refluxed 1 hour to reaction mixture then.At this moment, TLC and mass spectrum demonstration reaction is finished.Reaction mixture is with the ethyl acetate dilution and be cooled to 20 ℃.The throw out that obtains filters, and with more EtOAc washing and from MeOH/EtOAc/ charcoal/diatomite recrystallization, obtains B2 (1.531g, 98%); M.p (MeOH/EtOAc)〉300 ℃; ¹H NMR[(CD ₃) ₂SO] δ 12.86 (brs, 1H, N ⁺H), 10.90 (s, 1H, NH), 10.80 (br s, 1H, NH), 8.30-8.26 (m, 2H, ArH), 8.11-8.08 (m, 4H, ArH), 7.80-7.78 (br m, 3H, ArH ﹠amp; NH ₂), 7.56 (t, J=7.8Hz, 1H, ArH), 2.30 (s, 3H, CH ₃); Mass spectrum APCI ⁺365.

3-[(2-amino-6-methyl-4-pyrimidyl) amino]-N-(4-aminophenyl) benzamide hydrochloride salt (B3).(1.41g, 3.51mmol) suspension in (MeOH) 30 (mL) adds 10%Pd/C (20mg) and hydrogenation 5 hours under 45Hg mm to compd B 2.The reaction mixture filtration is evaporated to drying by Celite pad and with filtrate.Resistates is dissolved in the MeOH of a little volume, adds the MeOH that contains 1.25M HCl of 1ml then, stirred 10 minutes and be evaporated to drying.Resistates obtains B3 (758mg, 58%) from the MeOH/EtOAc recrystallization; M.p. (MeOH/EtOAc); ¹H NMR[(CD ₃) ₂SO] δ 12.70 (br, 1H, NH), 10.83 (br s, 1H, NH), 10.43 (s, 1H, NH), 9.70 (v br, 2H, NH ₂), 8.05-7.84 (br m, 2H, ArH), 7.85-7.75 (m, 4H, ArH ﹠amp; NH2), 7.55 (t, J=7.9Hz, 1H, ArH), 7.28 (d, J=8.7Hz, 2H, ArH), 6.24 (s, 1H, ArH), 2.30 (s, 3H, CH ₃); Mass spectrum APCI ⁺335.

3-[(2-amino-6-methyl-4-pyrimidyl) amino]-N-[4-(4-quinolyl amino) phenyl] benzamide dihydrochloride (compound F 17-hydroxy-corticosterone F1).(150mg is 0.37mmol) at EtOH (15mL) and H to compd B 3 ₂Solution among the O (7.5mL) adds the 4-chloroquinoline, and (73mg 0.45mmol) and stir, up to its dissolving, adds 2 dense HCl then.With reaction mixture refluxed 20 hours,, make its boiling and be cooled to 20 ℃ with the EtOAc dilution.The throw out that obtains is filtered and, obtain SN30962 (195mg, 96%), be yellow solid from MeOH/EtOAc/ charcoal/diatomite recrystallization. M.P (MeOH/EtOAc)〉300 ℃; ¹H NMR[(CD ₃) ₂SO] δ 13.72 (v br, 2H, 2 * N ⁺H), 10.91 (br s, 1H, NH), 10.96 (br s, 1H, NH), 10.56 (s, 1H, HN), 8.79 (d, J=8.5Hz, 1H, ArH), 7.50 (d, J=6.9Hz, 1H, ArH), 8.14-7.99 (m, 6H, ArH ﹠amp; NH2), 7.83 (m, 3H, ArH), 7.55 (t, J=7.9Hz, 1H, ArH), 7.49 (d, J=8.8Hz, 2H, ArH), 6.78 (d, J=6.9Hz, 1H, ArH), 6.24 (s, 1H, ArH), 2.30 (s, 3H, ArH), mass spectrum APCI ⁺476.

3-[(2,6-diamino-4-pyrimidyl) amino]-N-(4-nitrophenyl) benzamide hydrochloride salt (B5).(1.07g, 4.14mmol) with 4-chloro-2, (569mg 3.89mmol) is dissolved in warm cellosolvo (20mL) to 6-di-amino-pyrimidine (B4) with 3-amino-N-(4-nitrophenyl) benzamide (B1).Add two dense HCl and refluxed 20 hours to reaction mixture then, at this moment, TLC and mass spectrum show that reaction finishes.Reaction mixture is with the ethyl acetate dilution and be cooled to 20 ℃.The throw out that obtains is filtered, wash and from MeOH/EtOAc/ charcoal/diatomite recrystallization, obtain compd B 5 (824mg, 50%), m.p (MeOH/EtOAc) with more EtOAc 300 ℃; ¹H NMR[(CD ₃) ₂SO] δ 11.57 (br, 1H, N ⁺H), 10.84 (s, 1H, NH), 9.98 (s, 1H, NH), 8.30-8.26 (m, 2H, Ar), 8.10-8.07 (m, 2H, ArH), 8.06 (br s, 2H, NH ₂), 7.70 (d, J=7.2Hz, 1H, ArH), 7.53-7.49 (m, 3H, ArH), 7.38 (br s, 2H, NH ₂), mass spectrum APCI ⁺366.

N-(4-aminophenyl)-3-[(2,6-diamino-4-pyrimidyl) amino] benzamide hydrochloride salt (B6).(567mg 1.41mmol) adds 10%Pd/C (20mg) and hydrogenation 6 hours under 40Hgmm at (MeOH) suspension in (30mL) to compd B 5.The reaction mixture filtration is evaporated to drying by Celite pad and with filtrate.Resistates is dissolved in the MeOH of a little volume, adds the MeOH that contains 1.25M HCl of 1ml then, stir 10min and be evaporated to drying.Resistates obtains B6 (398mg, 76%) from the MeOH/EtOAc recrystallization; Mp (MeOH/EtOAc)〉300 ℃, ¹H NMR[(CD ₃) ₂SO] δ 10.37 (s, 1H, NH), 9.93 (s, 1H, NH), 7.92-7.80 (m, 4H, ArH ﹠amp; NH ₂), 7.68 (d, J=7.7Hz, 1H, ArH), 7.61 (brs, 2H, NH ₂), 7.49-7.45 (m, 3H, ArH), 7.24 (br d, J=8.7Hz, 2H, ArH), 5.45 (s, 1H, ArH), mass spectrum APCI ⁺336.

3-[(2,6-diamino-4-pyrimidyl) amino]-N-[4-(4-quinolyl amino) phenyl] benzamide dihydrochloride (compound F 17-hydroxy-corticosterone F2).(150mg is 0.37mmol) at EtOH (15mL) and H to compound B-26 ₂Solution among the O (7.5mL) adds the 4-chloroquinoline, and (73mg 0.45mmol) and stir, up to its dissolving, adds 2 dense HCl then.With reaction mixture refluxed 20 hours,, make its boiling and be cooled to 20 ℃ with the EtOAc dilution.The throw out that obtains filtered and at NH ₃Stir in the aqueous solution (20ml), to be converted into free alkali.The throw out that this is new filters and chromatogram purification (neutral Al ₂O ₃, 0-7% DCM/MeOH), obtain the pure free alkali of product.Use the MeOH that contains 1.25M HCl to be translated into HCl salt then, obtain compound F 17-hydroxy-corticosterone F2 (139mg, 70%); M.p (MeOH/EtOAc)〉300 ℃. ¹H NMR[(CD ₃) ₂SO] δ 14.25 (br, 1H, N ⁺H), 12.00 (br, 1H, N ⁺H), 10.95 (s, 1H, NH), 10.53 (s, 1H, NH), 9.93 (s, 1H, NH), 9.05 (d, J=8.5Hz, 1H, ArH), 7.51 (d, J=7.0Hz, 1H, ArH), 8.06-8.00 (m, 5H, ArH), 7.95-7.91 (br m, 2H, NH ₂), 7.81 (dd, J=7.6,1.4Hz, 1H, ArH), 7.71 (d, J=7.6Hz, 1H, ArH), 7.58 (s, 2H, NH ₂), 7.57-7.47 (m, 5H, ArH), 6.78 (d, J=7.0Hz, 1H, ArH), 6.78 (d, J=7.0Hz, 1H, ArH), 5.46 (s, 1H, ArH); HRMS (FAB ⁺) calculated value C ₂₆H ₂₃N ₈O (M ^{+ 1}) m/z463.1995, measured value 463.1992.

Embodiment GG.

4-[(2-amino-6-methyl-4-pyrimidyl) amino]-N-[4-(4-quinolyl amino) phenyl] preparation of benzamide dihydrochloride (compound GG1) and related compound (compound GG2, compound GG3, compound GG4, compound GG5)

(i) cellosolvo/H ⁺/ reflux; (ii) H ₂10% Pd/C/MeOH; The (iii) 4-chloroquinoline/EtOH/H of Qu Daiing ₂O/ refluxes

4-[(2-amino-6-methyl-4-pyrimidyl) amino]-N-(4-nitrophenyl) benzamide (5).With 4-amino-N-(4-nitrophenyl) benzamide (C1) (730mg, 2.84mmol) and 4-chloro-6-methyl-2-pyrimidyl amine (A8) (416mg, 2.84mmol) be dissolved in the warm cellosolvo (20mL), add two dense HCl to reaction mixture then, refluxed 40 minutes.At this moment TLC and MS demonstration reaction is finished.Reaction mixture is with the ethyl acetate dilution and be cooled to 20 ℃.The throw out that obtains is filtered and washs with more EtOAc, obtain solid product.Be suspended in it among MeOH and use NH ₃Aqueous solution alkalization and dilute with water.The throw out that obtains is filtered and drying, obtain C2 (671mg).Filtrate is concentrated, obtain the C2 (adding up to 92%) of other 276mg: mp (MeOH) 300 ℃; ¹H NMR[(CD ₃) ₂SO] δ 10.59 (s, 1H, NH), 9.37 (s, 1H, NH), 8.27-8.24 (m, 2H, ArH), 8.09-8.06 (m, 2H, ArH), 8.05-7.90 (m, 4H, ArH), 6.25 (s, 2H, NH ₂), 5.96 (s, 1H, ArH), 2.16 (s, 3H, CH ₃); Mass spectrum APCI ⁺365.

4-[(2-amino-6-methyl-4-pyrimidyl) amino]-N-(4-aminophenyl) benzamide (C3).(671mg, 1.84mmol) suspension in MeOH (50mL) was 40Hgmm hydrogenation (10% Pd/C50mg) 3 hours with C2.Product is precipitated out, and is white solid.With reaction mixture at (MeOH/HCl/H ₂O) stir and filter in (100mL/2mL/50mL) to remove the Pd/C resistates.Filtrate is evaporated to drying and, obtains C3 (740mg, 99%) from the MeOH/EtOAc crystallization; M.P. (MeOH/EtOAc)〉300 ℃; ¹H NMR[(CD ₃) ₂SO] δ 10.96 (bs, 1H, NH), 10.38 (s, 1H NH), 8.02-7.97 (m, 4H, ArH), 7.86 (d, J=8.8Hz, 2H, ArH), 7.30 (d, J=8.8Hz, 2H, ArH), 6.30 (d, J=0.6Hz, 1H, ArH), 2.33 (s, 3H, CH ₃).2 * NH ₂The signal of group is not observed; Mass spectrum APCI ⁺335.

4-[(2-amino-6-methyl-4-pyrimidyl) amino]-N-[4-(4-quinolyl amino) phenyl] benzamide dihydrochloride (compound GG1).(218mg is 0.54mmol) at EtOH (20mL) and H to C3 ₂Solution among the O (10mL) adds the 4-chloroquinoline, and (96mg 0.59mmol) and stir, up to its dissolving, adds 2 dense HCl then.With reaction mixture refluxed 3 hours,, make its boiling and be cooled to 20 ℃ with the EtOAc dilution.The throw out that obtains is filtered and, obtain compound GG1 (259mg98%), be yellow solid from MeOH/EtOAc/ charcoal/diatomite recrystallization.M.P (MeOH/EtOAc)〉300 ℃; ¹H NMR[(CD ₃) ₂SO] δ 13.75 (br.2H, 2 * N ⁺H), 11.07 (s, 1H, NH), 10.99 (s, 1H, NH), 10.52 (s, 1H, NH), 8.83 (d, J=8.4Hz, 1H, ArH), 8.50 (d, J=7.0Hz, 1H, ArH), 8.11-7.99 (m, 10H, ArH ﹠amp; NH ₂), 7.80 (dt, J=8.0,1.1Hz, 1H, ArH), 7.48 (d, J=8.9Hz, 2H, ArH), 6,78 (d, J=7.0Hz, 1H, ArH), 6.35 (s, 1H, ArH), 2.31 (s, 3H, CH ₃); ¹³C NMR[(CD ₃) ₂SO] δ 164.8,161.6,155.9,155.0,153.3,142.5,141.4,138.4,138.1,133.7,132.1,129.6,128.5 (2 * C), 126.8,125.8 (2 * C), 123.5,121.4 (2 * C), 120.2,120.1,116.9,99.6,94.3,18.4, there is a quaternary carbon to be difficult to observe; HRMS FAB ⁺Calculated value C ₂₇H ₂₅N ₇O (M ^{+ 1}) m/z, 462,2042 measured values 462.2047.

4-[(2-amino-6-methyl-4-pyrimidyl) amino]-N-{4-[(6-nitro-4-quinolyl) amino] phenyl } benzamide dihydrochloride (compound GG4).(200mg is 0.49mmol) at EtOH (20mL) and H to Compound C 3 ₂Solution among the O (10mL) adds 4-chloro-6-nitroquinoline, and (113mg 0.54mmol) and stir up to its dissolving, adds 2 dense HCl then.With reaction mixture refluxed 3 hours,, make its boiling and be cooled to 20 ℃ with the EtOAc dilution.The throw out that obtains is filtered and, obtain compound GG4 (247mg87%), be yellow solid from MeOH/EtOAc/ charcoal/diatomite recrystallization.295-300 ℃ of M.P (MeOH/EtOAc); ¹H NMR[(CD ₃) ₂SO] δ 13.50 (br, 2H, 2 * N ⁺H), 11.32 (br, 1H, NH), 11.00 (brs, 1H, NH), 10.51 (s, 1H, NH), 9.81 (d, J=2.2Hz, 1H, ArH), 8.69 (dd, J=9.3,2.2Hz, 1H, ArH), 8.59 (d, J=6.9Hz, 1H, ArH), 8.24 (d, J=9.3Hz, 1H, ArH), 8.06-7.98 (m, 7H, ArH﹠amp; NH ₂), 7.48 (d, J=8.9Hz, 2H, ArH), 6.90 (d, J=6.9Hz, 1H, ArH), 6.32 (s, 1H, ArH), 2.32 (d, J=0.4Hz, 3H, CH ₃); HRMS (FAB ⁺) calculated value C ₂₇H ₂₃N ₈O ₃(M ^{+ 1}) m/z 507.1893, measured value 507.1896.

4-[(2-amino-6-methyl-4-pyrimidyl) amino]-N-{4-[(6-amino-4-quinolyl base) amino] phenyl } benzamide dihydrochloride (compound GG3).(140mg, 0.24mmol) suspension in (MeOH) 30 (mL) adds 10% Pd/C and hydrogenation 1 hour under 45Hgmm to SN 31001.The reaction mixture filtration is evaporated to drying by Celite pad and with filtrate.Resistates is dissolved in the MeOH of a little volume, adds the MeOH that contains 1.25M HCl of 1ml then, stir 10min and be evaporated to drying.Resistates obtains compound GG3 (107mg, 81%) from the MeOH/EtOAc recrystallization; M.p. (MeOH/EtOAc)〉300 ℃ ¹H NMR[(CD ₃) ₂SO] δ 14.13 (d, J=5.6Hz, 1H, N ⁺H), 12.95 (s, 1H, N ⁺H), 10.97 (s, 1H, NH), and 10.44, (s, 1H, NH), 10.20 (s, 1H, NH), 8.22 (t, J=6.5Hz, 1H, ArH), and 8.05-7.96 (m, 7H, ArH), 7.79 (d, J=9.1Hz, 1H, ArH), 7.49 (d, J=1.9Hz, 1H, ArH), 7.43-7.38 (m, 3H, ArH), 6.66 (d, J=6.8Hz, ArH), 6.30 (s, 1H, ArH), 2.24 (d, J=0.6Hz, 3H, CH ₃); HRMS (FAB ⁺) calculated value C ₂₇H ₂₅N ₈O (M ⁺1) m/z 477.2151, measured value 477.2153.

4-[(2-amino-6-methyl-4-pyrimidyl) amino]-N-{4-[(7-nitro-4-quinolyl) amino] phenyl } benzamide dihydrochloride (compound GG5).(235mg is 0.58mmol) at EtOH (20mL) and H to C3 ₂Solution among the O (10mL) adds 4-chloro-7-nitroquinoline, and (132mg 0.63mmol) and stir up to its dissolving, adds 2 dense HCl then.With reaction mixture refluxed 5 hours,, make its boiling and be cooled to 20 ℃ with the EtOAc dilution.The throw out that obtains is filtered and, obtain compound GG5 (317mg 94%), be yellow solid from MeOH/EtOAc/ charcoal/diatomite recrystallization.M.p (MeOH/EtOAc)〉300 ℃; ¹HNMR[(CD ₃) ₂SO] δ 12.9 (br, 1H, N ⁺H), 10.81 (brs, 2H, 2 * NH), 10.44 (s, 1H, NH), 8.93 (d, J=9.3Hz, 1H, ArH), 8.81 (d, J=1.9Hz, 1H, ArH), 8.66 (d, J=6.6Hz, 1H, ArH), 8.46 (d, J=8.9Hz, 1H, ArH), 8.05-7.98 (m, 8H, ArH ﹠amp; NH ₂), 7.47 (d, J=8.8Hz, 1H, ArH), 6.94 (d, J=6.6Hz, 1H, ArH), 6.26 (s, 1H, ArH), 2.32 (s, 3H, CH ₃); HRMS (FAB ⁺) calculated value C ₂₇H ₂₃N ₈O ₃(M ^{+ 1}) m/z 507.1893, measured value 507.1885.

4-[(2-amino-6-methyl-4-pyrimidyl) amino]-N-{4-[(7-amino-4-quinolyl base) amino] phenyl } benzamide dihydrochloride (compound GG2).(150mg, 0.25mmol) suspension in (MeOH) 30 (mL) adds 10% Pd/C (20mg) and hydrogenation 1 hour under 45Hgmm to SN 31002.The reaction mixture filtration is evaporated to drying by Celite pad and with filtrate.Resistates is dissolved in the MeOH of a little volume, adds the MeOH that contains 1.25M HCl of 1ml then, stir 10min and be evaporated to drying.Resistates obtains compound GG2 (141mg, 100%), 257-262 ℃ of m.p (MeOH/EtOAc) from the MeOH/EtOAc recrystallization. ¹H NMR[(CD ₃) ₂SO] δ 13.45 (br, 2H, 2 * N ⁺H), 10.33 (s, 1H, NH), 10.30 (s, 1H, NH), 10.00 (br, 1H, NH), 8.33 (d, J=9.4Hz, 1H, ArH), 8.14 (d, J=7.0Hz, 1H, ArH), 3.19-7.92 (m, 6H, ArH), 7.39 (d, J=8.9Hz, 2H, ArH), and 7.04-7.01 (br m, 3H, ArH), 6.86 (d, J=2.2Hz, 1H, ArH), 6.67 (br s, 2H, NH ₂), 6.43 (d, J=7.0Hz, 1H, ArH﹠amp; NH ₂)), 6.13 (s, 1H, ArH), 2.22 (s, 3H, CH ₃); HRMS (FAB ⁺) calculated value C ₂₇H ₂₅N ₈O (M ^{+ 1}) m/z 477.2151, measured value 477.2153.

Embodiment HH.

4-[(2-amino-6-methyl-4-pyrimidyl) amino]-preparation of N-(4-{[6-(dimethylamino)-4-quinolyl] amino } phenyl) benzamide dihydrochloride (compound H H)

(i) cellosolvo/H+/backflow; (ii) A3/EDC/DMAP/N-methyl-2-pyrrolidone

4-[(2-amino-6-methyl-4-pyrimidyl) amino] benzoate hydrochlorate (D2).With para-amino benzoic acid (D1) (1.48g, 10.8mmol) and 4-chloro-6-methyl-2-PYRIMITHAMINE (A8) (1.60g 11.9mmol) is dissolved in warm cellosolvo (20mL), adds two dense HCl then and refluxes 1 hour.Reaction mixture is cooled to 20 ℃ and the throw out that obtains filtered, washs with more cellosolvo and EtOAc.Solid is seethed with excitement in MeOH, and cooling and filtration with more MeOH washing and dry, obtain D2 (2.89g, 95%); M.p. (MeOH)〉300 ℃; ¹H NMR[(CD ₃) ₂SO] δ 12.93 (br, 1H, N ⁺H or COOH), 12.80 (br, 1H, N ⁺H or COOH), 10.87 (s, 1H, NH), 7.92 (s, 6H, ArH ﹠amp; NH ₂)), 6.27 (s, 1H, ArH), 2.32 (s, 3H, CH ₃); Mass spectrum APCI ⁺245.

4-[(2-amino-6-methyl-4-pyrimidyl) amino]-N-(4-([6-(dimethylamino)-4-quinolyl] amino } phenyl) benzamide dihydrochloride (compound H H).With Compound D 2 (111mg, 0.4mmol), EDCI (155mg, 0.72mmol) and DMAP (174mg 1.44mmol) stirs 5min at 20 ℃ in N-Methyl pyrrolidone (5mL).Add N then ⁴-(4-aminophenyl)-N ⁶, N ⁶-dimethyl-4, (100mg 0.36mmol) and with reaction mixture stirred 20 hours at 20 ℃ 6-quinoline diamines.Reaction mixture dilute with water and stirring.The throw out that obtains is filtered, wash with water and dry air.This solid is dissolved in the MeOH of heat, seethes with excitement and filter and pass through Celite pad with charcoal.Filtrate is evaporated to drying and the resistates that obtains is dissolved in MeOH (10mL), and adding contains the MeOH (1mL) of 1.25M HCl and stirred 10 minutes.With solvent evaporation and with resistates from the MeOH/EtOAc recrystallization, obtain compound H H (135mg, 65%), be yellow solid; M.p. (MeOH/EtOAc)〉300 ℃; ¹H NMR[(CD ₃) ₂SO] δ 14.10 (br, 1H, N ⁺H), 12.75 (br, 1H, N ⁺H), 10.85 (br, 1H, NH), 10.45 (s, 1H, NH), 10.40 (s, 1H, NH), 8.27 (d, J=6.8Hz, 1H, ArH), 8.08-7.99 (m, 6H, ArH), 7.88 (d, J=9.4Hz, 1H, ArH), 7.65 (dd, J=9.4,2.5Hz, 1H, ArH), 7.54 (d, J=2.4Hz, 1H, ArH), 7.45 (d, J=8.9Hz, 2H, ArH), 6.67 (d, J=6.8Hz, 1H, ArH), 6.27 (s, 1H, ArH), 3.12[s, 6H, N (CH ₃) ₂], 2.32 (s, 3H, CH ₃); Mass spectrum APCI ⁺505.

Example II.

N-[3-(2-amino-6-methylpyrimidine-4-base amino) phenyl]-preparation of 4-(quinolyl-4 amino) benzamide hydrochloride salt (Compound I I)

(i) 3-N-methyl-p-nitroaniline/cellosolvo/cHc/ refluxes; The (ii) EtOH/H of Fe powder/(2:1) ₂O, 2%v/vHCl/ refluxes; (iii) 4-nitrobenzoyl chloride/pyridine/EtOH/ refluxes; (iv) 2:1 EtOH/H2O/cHC//backflow

6-methyl-N ⁴-(3-nitrophenyl) pyrimidine-2,4-diamines (E1).To 2-amino-4-chloro-6-methyl-pyrimidine (A8) [10.04g, 69.93mmol] and 3-N-methyl-p-nitroaniline 99.95g, 72.02mmol) suspension in cellosolvo (300mL) adds dense HCl (30mL), and the solution that obtains was refluxed～16 hours.After this, make the reaction mixture cool to room temperature, use salt solution and H then ₂The O dilution.The suspension filtered that obtains is passed through Celite pad, and the solid matter that will so collect is dissolved in MeOH more then, and filters and pass through Celite pad.The solvent decompression is removed, obtain diamines E1, be unbodied beige solid, it is without using through single step purification: ¹H NMR (400MHz, DMSO): δ 12.97 (brs, 1H, pyrimidyl-N ⁺-H), 11.03 (s, 1H, ArNHAr), 8.52 (s, 1H, ArH), 8.31 (d, J=7.83Hz, 1H, ArH), 7.97 (ddd, J=8.20,2.19,0.72Hz, 1H, ArH), 7.83 (v br s, 2H, ArNH ₂), 7.66 (t, J=8.21Hz, 1H, ArH), 6.25 (s, 1H, ArH), 2.31 (s, 3H, ArCH ₃); LCMS (APCI ⁺): 246 (100%).

N ⁴-(3-aminophenyl)-6-methylpyrimidine-2,4-diamines (E2).To at the diamines E1 that refluxes at 2:1 EtOH:H ₂Suspension among the O (500mL) sequentially adds Fe powder (15.6g is 4 molar equivalents with respect to 1) and dense HCl (10mL), and the mixture backflow that obtains is spent the night.After this, Celite pad is passed through in the reaction mixture filtration of heat, and the solvent decompression is removed.Resistates is dissolved in again the H of heat ₂O, and make the suspension filtered that obtains pass through Celite pad.The solvent decompression is removed, and resistates is passed through three MeOH-azeotropic cyclic dryings.The resistates that obtains obtains diamines E2 (5.36g, 30% fromA8) from acidifying MeOH:EtOAc redeposition, is unbodied brown solid; ¹H NMR (400MHz, DMSO): δ 12.90 (vvbrs, 1H, pyrimidyl-N ⁺-H), 11.00 (br s, 1H, ArH), 9.75 (v v br s, 2H, ArNH ₂), 8.20-7.50 (m, 4H, ArH ﹠amp; ArNH ₂), 7.37 (t, J=8.05Hz, 1H, ArH), 6.98 (d, J=7.82Hz, ArH, 1H), 6.26 (s, 1H, ArH), 2.29 (s, 3H, ArCH ₃); LCMS (APCI ⁺): 216 (100%), 217 (40%).

N-[3-(2-amino-6-methylpyrimidine-4-base is amino) phenyl]-4-nitrobenzamide (E3).To diamines E2 (5.25g, 20.85mmol) and anhydrous pyridine (8.40mL, 104.26mmol) suspension in no Shui diox (200mL) adds the 4-nitrobenzoyl chloride (10.79g 58.13mmol), and refluxes the solution that obtains～14 hours.After this, with the reaction mixture cool to room temperature, then by adding the ammonia soln alkalization.The solution H that obtains ₂The O dilution, and, obtain acid amides E3 by filtering the throw out that collection obtains, be unbodied yellow-brown solid (7.89g, 94%) [a spot of this material is used for characterizing from acidifying MeOH:EtOAc redeposition, and with the material of major portion without using] through single step purification; ¹H NMR (400MHz, DMSO): δ 12.99 (br s, 1H, pyrimidyl-N ⁺-H), 10.82 (s, 1H, ArNHAr), 10.76 (br s, 1H, ArC (O) NHAr), 8.35 (dd, J=6.92,1.99Hz, 2H, ArH), 8.27 (d, J=6.92,1.99Hz, 2H, ArH), 7.90 (v v v br s, 2H, ArNH ₂), 8.20-7.50 (m, 4H, ArH ﹠amp; ArNH ₂), 7.45-7.10 (m, 2H, ArH), 6.25 (s, 1H, ArH), 2.28 (s, 3H, ArCH ₃); LCMS (APCI ⁺): 365 (100%).

4-amino-N-[3-(2-amino-6-methylpyrimidine-4-base is amino) phenyl] benzamide (E4).To (7.89g is 20.00mmol) at 2:1 EtOH:H at the acid amides E3 that refluxes ₂Suspension among the O (500mL) sequentially add the Fe powder (4.40g, 79mmol) and dense HCl (2%v/v 10mL), and refluxes the mixture that obtains～14 hours.After this, Celite pad is passed through in the reaction mixture filtration of heat, and the solvent decompression is removed.Resistates is dissolved in hot H again ₂O, and make the suspension filtered that obtains pass through Celite pad.The solvent decompression is removed, and resistates is passed through three MeOH-azeotropic cyclic dryings.The resistates that obtains obtains amine E4 from acidifying MeOH:EtOAc redeposition, is unbodied brown solid (0.95g, 12%); ¹H NMR (400MHz, DMSO): δ 12.71 (br s, 1H, pyrimidyl-N ⁺-H), 10.55 (br s, 1H, ArC (O) NHAr), 9.95 (s, 1H, ArNHAr), 8.10-7.40 (m, 7H, ArH ﹠amp; ArNH ₂), 7.32 (m, 1H, ArH), 6.77 (d, J=8.30Hz, 2H, ArH), 6.18 (s, 1H, ArH), 2.28 (s, 3H, ArCH ₃); LCMS (APCI ⁺): 335 (100%).

N-[3-(2-amino-6-methylpyrimidine-4-base is amino) phenyl]-4-(quinolyl-4 amino) benzamide hydrochloride salt (Compound I I).(0.26g is 0.63mmol) at 1:2EtOH:H to amine E4 ₂Solution among the O (30mL) sequentially add the 4-chloroquinoline (0.62g, 3.78mmol) and dense HCl (0.17mL 5.67mmol), and refluxes the mixture that obtains that (logical TLC followed the tracks of reaction process, used N-BuOH:H in 3 hours ₂O: the last phase wash-out of the 5:4:1 mixture of acetate; Product Rf=0.43 is using KMNO ₄After the dyeing is yellow spotting).After this, the solvent decompression is removed, and resistates is passed through three MeOH-azeotropic cyclic dryings.Resistates obtains Compound I I from the MeOH:EtOAc redeposition, for unbodied lemon yellow-yellow solid (0.27g, 79%); Mp251-255 ℃ (powder-jelly); ¹H NMR (400MHz, DMSO): δ 14.78 (v brs, 1H, quinolyl-N ⁺-H), 12.79 (v br s, 1H, pyrimidyl-N ⁺-H), 11.14 (s, 1H, ArNHAr), 10.64 (br s, 1H, ArC (O) NHAr), 10.52 (s, 1H, ArNHAr), 8.87 (d, J=8.44Hz, 1H, ArH), 8.61 (d, J=6.94Hz, 1H, ArH), 8.30-7.45 (m, 12H, ArH ﹠amp; ArNH ₂), 7.38 (t, J=8.07Hz, 1H, ArH), 7.00 (d, J=6.94Hz, 1H, ArH), 6.22 (s, 1H ArH), 2.29 (s, 3H, ArCH ₃); LCMS (APCI ⁺): 463 (100%); HPLC:95.4%.

Embodiment JJ.

(E)-N-[3-(1-{ (diamino methene base) hydrazono-} ethyl) phenyl]-4-[6-(dimethylamino) quinolyl-4 amino] benzamide dihydrochloride (compound JJ1) and related compound (compound JJ2) synthetic

(i) guanidine sulfate/MeOH/c.HCl/ refluxed/14 hours; (ii) Fe powder, cat.c.HCl, 2:1 EtOH:H ₂O refluxes 14 hours; (iii) pyridine ， diox refluxes 17 hours; (iv) 4-Delagil/1:2 EtOH/H ₂O/c.HCl/ refluxed/20 hours; (vi) 4-chloro-6-(dimethylamino) quinoline/1:2 EtOH/H ₂O/c.HCl/ refluxes, 40 hours

(E)-N-[4-(1-{ diamino methene base } hydrazono-) ethyl]-3-oil of mirbane hydrochloride (F2).To aminoguanidine sulfate (21.25g, 80.41mmol) and 3-nitro-acetophenone (F1) (13.53g, 81.93mmol) solution in MeOH (400mL) adds dense HCl (2.44mL 80.42mmol) and with the mixture that obtains refluxed～14 hours.Remove from the white suspension that obtains and to desolvate, obtain diamines F2, be unbodied white solid, it is without using through single step purification; ¹H NMR (400MHz, DMSO): δ 11.00 (v v br s, 1H, hydrazoNy1-N ⁺-H), 8.59 (t, J=1.94Hz, 1H, ArH), 8.39 (d, J=7.87Hz, 1H, ArH), 8.19 (ddd, J=8.14,2.18,0.82Hz, 1H, ArH), 7.83 (brs, 4H ,=C (NH ₂) ₂), 7.66 (t, J=8.05Hz, 1H, ArH), 2.40 (s, 3H, Ar (CH ₃)=N-); LCMS (APCI ⁺): 222 (100%).

(E)-N-[4-(1-{ diamino methene base } hydrazono-) ethyl]-3-aniline dihydrochloride (F3).To at the diamines F2 that refluxes at 2:1 EtOH:H ₂Suspension among the O (500mL) sequentially adds Fe powder (17.87g, 320.00mmol, 4 molar equivalents) and dense HCl (10mL), and the mixture that obtains was refluxed 14 hours.After this, the reaction mixture of heat is filtered by Celite pad, and the solvent decompression is removed.Resistates is dissolved in hot H again ₂O, and make the suspension filtered that obtains pass through Celite pad.The solvent decompression is removed, and resistates is passed through three MeOH-azeotropic cyclic dryings.The resistates that obtains obtains triamine F3 (10.78g, 51% over two steps) from acidifying MeOH/EtOAc redeposition, is unbodied white solid; ¹H NMR (400MHz, DMSO): δ 11.35 (s, 1H, hydrazoNyl-N ⁺-H), 9.40 (vv br s, 3H, ArNH ₃ ⁺), 7.84 (br m, 5H, ArH ﹠amp;=C (NH ₂) ₂), 7.75 (s, 1H, ArH), 7.43 (t, J=7.91Hz, 1H, ArH), 7.27 (d, J=8.27Hz, 1H, ArH), 2.34 (s, 3H, Ar (CH ₃)=N-); LCMS (APCI ⁺): 192 (100%).

(E)-N-[3-(1-{ (diamino methene base) hydrazono-} ethyl) phenyl]-4-nitrobenzamide dihydrochloride (F4).To triamine F3 (5.15g, 19.49mmol) and anhydrous pyridine (7.85mL, 97.45mmol) suspension in no Shui diox (300mL) adds the 4-nitrobenzoyl chloride (10.52g 56.69mmol), and refluxes the mixture that obtains 17 hours.After this, with the reaction mixture cool to room temperature, then by adding the ammonia soln alkalization.The solution H that obtains ₂The O dilution, and, obtain acid amides F4 (4.21g, 51%) by filtering the throw out that collection obtains, be unbodied butyraceous yellow solid; ¹H NMR (400MHz, DMSO): δ 11.09 (s, 1H, hydrazoNyl-N ⁺-H), 10.68 (s, 1H, ArC (O) NHAr), 8.39 (d, J=9.22Hz, 1H, ArH), 8.29 (m, 3H, ArH), 7.89 (d, J=8.07Hz, 1H, ArH), 7.82 (d, J=7.89Hz, 1H, ArH), 7.73 (br s, 4H ,=C (NH ₂) ₂), 7.44 (t, J=7.98Hz, 1H, ArH), 2.34 (s, 3H, Ar (CH ₃)=N-); LCMS (APCI ⁺): 341 (100%).

(E)-and 4-amino-N-[3-(1-{ (diamino methene base) hydrazono-} ethyl) phenyl] benzamide dihydrochloride (F5).To (2.02g is 5.36mmol) at 2:1EtOH:H at the acid amides F4 that refluxes ₂Suspension among the O (100mL) sequentially adds the Fe powder, and (1.20g 21.44mmol) and dense HCl (2mL), and made the suspension returning that obtains 14 hours.After this, Celite pad is passed through in the reaction mixture filtration of heat, and the solvent decompression is removed.Resistates is dissolved in hot H again ₂O, and make the suspension filtered that obtains pass through Celite pad.The solvent decompression is removed, and resistates is passed through three MeOH-azeotropic cyclic dryings.The resistates that obtains obtains triamine F5 (2.18g, quantitative yield) from acidifying MeOH redeposition, is unbodied cream-colored solid; ¹H NMR (400MHz, DMSO): δ 11.30 (s, 1H, hydrazone group-N ⁺-H), 10.05 (s, 1H, ArC (O) NHAr), 8.21 (s, 1H, ArH), 8.20-7.55 (m, 8H, ArH ﹠amp;=C (NH ₂) ₂), 7.36 (t, J=7.90Hz, 1H, ArH), 6.91 (d, J=7.96Hz, 1H, ArH), 5.30 (vbrs, 3H, ArNH ₃ ⁺), 2.35 (s, 3H, Ar (CH ₃)=N-); LCMS (APCI ⁺): 311 (100%).

(E)-N-[3-(1-{ (diamino methene base) hydrazono-} ethyl) phenyl]-4-[6-(dimethylamino) quinolyl-4 amino] benzamide dihydrochloride (compound JJ1).To triamine F5 (0.22g, 0.59mmol) 1:2 EtOH:H ₂Solution among the O (60mL) sequentially add 6-dimethylamino-4-chloroquinoline (0.13g, 0.64mmol) and dense HCl (0.16mL 5.32mmol), and refluxes the solution that obtains that (logical TLC followed the tracks of reaction process, used N-BuOH:H in 20 hours ₂O: the last phase wash-out of the 5:4:1 mixture of acetate).After this, the solvent decompression is removed, and resistates is passed through two MeOH-azeotropic cyclic dryings.Resistates obtains compound JJ1 (61mg, 19%) from the MeOH:EtOAc redeposition, is unbodied dun solid; Mp239-243 ℃ (powder → jelly), 278-282 ℃ (jelly → liquid); ¹H NMR (400MHz, DMSO): δ 14.44 (s, 1H, quinolyl-N ⁺-H), 11.20 (s, 1H, hydrazono--N ⁺-H), 10.58 (s, 1H, ArC (O) NHAr), 10.46 (s, 1H, ArNHAr), 8.35 (d, J=6.71Hz, 1H, ArH), 8.26 (t, J=1.73Hz, 1H, ArH), 8.22 (d, J=8.61Hz, 3H, ArH), 7.95 (m, 2H, ArH), 7.79 (m, 5H, ArH﹠amp;=C (NH ₂) ₂), 7.69 (d, J=2.52Hz, 1H, ArH), 7.66 (d, J=8.61Hz, 2H, ArH), 7.58 (d, J=2.43Hz, 1H, ArH), 7.43 (t, J=7.98Hz, 1H, ArH), 6.95 (d, J=6.71Hz, 1H, ArH), 3.15 (s, 6H, ArN (CH ₃) ₂), 2.37 (s, 3H, Ar (CH ₃)=N-); LCMS (APCI ⁺): 482 (100%); HPLC:95.8%.

(E)-N-[3-(1-{ (diamino methene base) hydrazono-} ethyl) phenyl]-4-(quinolyl-4 amino) benzamide dihydrochloride (compound JJ2).(0.35g is 0.91mmol) at 1:2 EtOH:H to triamine F5 ₂Solution among the O (30mL) sequentially add the 4-chloroquinoline (0.64g, 3.90mmol) and dense HCl (0.25mL 8.24mmol), and refluxes the solution that obtains and (followed the tracks of reaction process by TLC, and used N-BuOH:H in 20 hours ₂O: the last phase wash-out of the 5:4:1 mixture of acetate; Product R _f=0.51, using KMNO ₄After the dyeing is yellow spotting).After this, the solvent decompression is removed, and resistates is passed through two MeOH-azeotropic cyclic dryings.Resistates obtains compound JJ2 (91mg, 20%) from the MeOH:EtOAc redeposition, is light yellow amorphous solid; Mp 240-244 ℃ (powder → jelly), 264-268 ℃ (jelly → liquid); ¹H NMR (400MHz, DMSO): δ 14.67 (br s, 1H, quinolyl-N ⁺-H), 11.19 (s, 1H, hydrazono--N ⁺-H), 11.07 (s, 1H, ArC (O) NHAr), 10.48 (s, 1H, ArNHAr), 8.72 (d, J=8.52Hz, 1H, ArH), 8.61 (d, J=6.90Hz, 1H, ArH), 8.24 (m, 3H, ArH), 8.08 (m, 2H, ArH), 7.95 (d, J=8.10Hz, 1H, ArH), 7.82 (m, 6H, ArH ﹠amp;=C (NH ₂) ₂), 7.68 (d, J=8.38Hz, 2H, ArH), 7.43 (t, J=7.97Hz, 1H, ArH), 7.01 (d, J=6.90Hz, 1H, ArH), 2.37 (s, 3H, Ar (CH ₃) ₂=N-); LCMS (APCI ⁺): 438 (100%); HPLC:96.4%.

Embodiment KK.

N-[4-(2-amino-6-methylpyrimidine-4-base amino) phenyl]-4-(6,7,8-quinolinium trifluoroacetate-4-base is amino) benzamide dihydrochloride (compound K K) synthetic

Reflux=refluxes

6,7,8-three fluoro-4-oxos-1,4-dihydroquinoline-3-carboxylic acid (G2).(3.82g's quinolinium trifluoroacetate ketone ester G1, solution 14.09mmol) refluxed 14 hours in 1M NaOH, then cool to room temperature.With solution 1M NaOH acidifying, and with the suspension filtered that obtains, obtain quinolinium trifluoroacetate ketone acid G2 (3.31g, 97%), be unbodied white solid, it is without using through single step purification; Mp 264-268 ℃; ¹H NMR (400MHz, DMSO): δ 14.20 (v br s, 2H, quinolyl-N ⁺-H ﹠amp; ArCO ₂H), 8.71 (s, 1H, ArH), 8.08 (ddd, J _H-F=10.17,7.77,2.23Hz, 1H, ArH); LCMS (APCI ⁺): 244 (100%).

6,7,8-quinolinium trifluoroacetate-4 (1H)-ketone (G3).(1.32g, solution 5.43mmol) refluxed 30 minutes in diphenyl ether (100mL) with quinolinium trifluoroacetate ketone acid G2.After this, the reaction mixture of heat is poured in the hexane carefully, and makes the suspension cool to room temperature that obtains.With suspension filtered, obtain quinolinium trifluoroacetate ketone G3 (two batch of materials add up to 1.32g, quantitative yield), be very thin unbodied pale solid, it is without using through single step purification; ¹H NMR (400MHz, DMSO): δ 12.14 (s, 1H, ArOH), 7.91 (dd, J=6.73,6.36Hz, 1H, ArH), 7.80 (ddd, J _H-F=10.46,8.11,2.14Hz, 1H, ArH), 6.10 (d, J=7.43Hz, 1H, ArH); LCMS (APCI ⁺): 200 (100%).

4-chloro-6,7,8-quinolinium trifluoroacetate (G4).(1.30g, solution 6.53mmol) is at POCl with quinolinone G3 ₃Refluxed 1.3 hours (50mL), then with excessive POCl ₃Decompression is removed.Resistates is dissolved in CH again ₂Cl ₂, and by adding ammonia soln with the solution alkalization that obtains.The solution CH that obtains ₂Cl ₂Extract, and the organic extraction that merges is sequentially used H ₂O and salt water washing, and use MgSO ₄Dry.The solvent decompression is removed, and, use 100% CH the purified by flash chromatography on the resistates silica gel ₂Cl ₂→ 1% MeOH:CH ₂Cl ₂→ 10%MeOH:CH ₂Cl ₂Wash-out), obtains quinolinium trifluoroacetate G4 (0.76g, 54%), be white crystalline solid; Mp 119-121 ℃; ¹H NMR (400MHz, DMSO): δ 8.93 (d, J=4.73Hz, 1H, ArH), 8.08 (ddd, J _H-F=10.23,7.92,2.30Hz, 1H, ArH), 7.95 (d, J=4.73Hz, 1H, ArH); LCMS (APCI ⁺): 218 (100%), 220 (100%).

N-[4-(2-amino-6-methylpyrimidine-4-base is amino) phenyl]-4-(6,7,8-quinolinium trifluoroacetate-4-base is amino) benzamide dihydrochloride (compound K K).(0.19g, 0.46mmol) solution in anhydrous MeOH (20mL) sequentially adds quinolinium trifluoroacetate G4 (0.22g 1.03mmol) and dense HCl (～3), and refluxes the solution that obtains and (followed the tracks of reaction process by TLC, and used N-BuOH:H in 24 hours to amine G5 ₂O: the last phase wash-out of the 5:4:1 mixture of acetate; Product Rf=0.56 is using KMNO ₄After the dyeing is yellow spotting.It has the Rf identical with G5, yet, confirm the consumption of G5 by lcms analysis).After this, the solvent decompression is removed, and resistates is passed through three MeOH-azeotropic cyclic dryings.Resistates sequentially with MeOH, EtOAc and hexane grind, and be dry under high vacuum then, obtains compound K K (two batch of materials add up to 214mg, 79%), is unbodied pale asphyxia/lemon-yellow solid; Mp〉280 ℃; ¹H NMR (400MHz, DMSO): δ 12.70 (br s, 1H, pyrimidyl-N-H ⁺), 10.77 (br s, 1H, ArC (O) NHAr), 10.66 (s, 1H, ArNHAr), 10.39 (s, 1H, ArNHAr), 8.84 (m, 1H, ArH), 8.59 (d, J=6.35Hz, 1H, ArH), 8.14 (d, J=8.62Hz, 2H, ArH), 8.06-7.64 (m, 6H, ArH ﹠amp; ArNH ₂), 7.61 (d, J=8.62Hz, 2H, ArH), 7.11 (d, J=6.35Hz, 1H, ArH), 6.19 (s, 1H, ArH), 2.28 (s, 3H, ArCH ₃) [quinolyl-N ⁺-H is invisible]; LCMS (APCI ⁺): 517 (100%), 518 (40%); HPLC:98.9%.

Embodiment LL.

N-[4-(2-amino-6-methylpyrimidine-4-base amino) phenyl]-4-(6-chloroquinoline-4-base is amino) benzamide (compound L L) synthetic

4,6-dichloroquinoline (H2).6-chloro-4-quinolinone (H1) (1.48g, POCl 8.22mmol) ₃(50mL) solution refluxed 3.5 hours, then with excessive POCl ₃Decompression is removed.Resistates is dissolved in CH again ₂Cl ₂, and by adding ammonia soln with the solution alkalization that obtains.The solution CH that obtains ₂Cl ₂Extract, and the organic extraction that merges is sequentially used H ₂O and salt water washing, and use MgSO ₄Dry.The solvent decompression is removed, obtain dichloroquinoline H2 (1.54g, 95%), be unbodied butteriness white solid; Mp 101-103 ℃; R _f=0.83 (5%MeOH:CH ₂Cl ₂); ¹H NMR (400MHz, DMSO): δ 8.88 (d, J=4.73Hz, 1H, ArH), 8.21 (d, J=2.33Hz, 1H, ArH), 8.14 (d, J=8.99Hz, 1H, ArH), 7.91 (dd, J=8.99,2.33Hz, 1H, ArH), 7.84 (d, J=4.73Hz, 1H, ArH); LCMS (APCI ⁺): 198 (100%), 200 (80%).

N-[4-(2-amino-6-methylpyrimidine-4-base is amino) phenyl]-4-(6-chloroquinoline-4-base is amino) benzamide (compound L L).(0.18g, 0.44mmol) solution in anhydrous MeOH (40mL) sequentially adds dichloroquinoline H2 (0.17g 0.88mmol) and dense HCl (several), and refluxes the mixture that obtains 2 hours to amine G5.TLC afterwards analyzes and (uses N-BuOH:H ₂O: the last phase wash-out of the 5:4:1 mixture of acetate) but show that H2 has consumed some E4 residues, therefore added at 3 hours another part H2 (0.17g, 0.88mmol).The mixture that obtains was refluxed 14 hours, analyzes by TLC and do not observe variation, the solvent decompression is removed and with resistates by three MeOH-azeotropic cyclic dryings.Resistates and MeOH grind, and be dry under high vacuum then, obtains compound L L (2 batch of materials add up to 0.21g, 83%), is unbodied yellow solid; Mp 250-254 ℃; ¹H NMR (400MHz, DMSO): δ 14.80 (v br s, 1H, pyrimidyl-N-H ⁺), 12.62 (br s, 1H, quinolyl-N ⁺-H), 11.07 (s, 1H, ArC (O) NHAr), 10.62 (s, 1H, ArNHAr), 10.44 (s, 1H, ArNHAr), 9.01 (d, J=1.50Hz, 1H, ArH), 8.63 (d, J=6.90Hz, 1H, ArH), 8.13 (m, 4H, ArH), 8.05-7.45 (m, 8H, ArH ﹠amp; ArNH ₂), 7.04 (d, J=6.90Hz, 1H, ArH), 6.18 (s, 1H, ArH), 2.28 (s, 3H, ArCH ₃); LCMS (APCI ⁺): 497 (100%), 499 (60%); HPLC:99.7%.

Embodiment MM.

N-[4-(pyridin-4-yl amino) phenyl]-4-(quinolyl-4 amino) benzamide dihydrochloride (compound MM) synthetic

(i) 4-chloroquinoline/c.HCl, 1:10MeOH:EtOH refluxes

N-[4-(pyridin-4-yl amino) phenyl]-4-(quinolyl-4 amino) benzamide dihydrochloride (compound MM).To amine I1 (0.45g, 1.18mmol) suspension in 1:10 MeOH:EtOH sequentially add the 4-chloroquinoline (0.97g, 5.91mmol), dense HCl (3mL) and H ₂O (10mL), and the mixture that obtains refluxed (logical TLC followed the tracks of reaction process, used N-BuOH:H in～14 hours ₂O: the last phase wash-out of the 5:4:1 mixture of acetate; Product R _f=0.30, using KMNO ₄After the dyeing is yellow spotting).Afterwards, the solvent decompression is removed, and resistates is passed through three MeOH-azeotropic cyclic dryings.Resistates is from the MeOH:EtOAc redeposition, and is further purified by preparation property HPLC, obtains compound MM (0.14g, 32% brsm), for unbodied lemon yellow-yellow solid; Mp:148-151 ℃ (powder → jelly), 171-174 ℃ (jelly → liquid); ¹H NMR (400MHz, DMSO): δ 13.97 (brs, 2H, pyridyl-N ⁺-H﹠amp; Quinolyl-N ⁺-H), 10.83 (brs, 1H, ArC (O) NHAr), 10.49 (s, 1H, ArNHAr), 10.44 (s, 1H, ArNHAr), 8.71 (d, J=8.47Hz, 1H, ArH), 8.63 (d, J=6.81Hz, 1H, ArH), 8.27 (d, J=7.14Hz, 2H, ArH), 8.17 (d, J=8.51Hz, 2H, ArH), 8.05 (m, 2H, ArH), 7.94 (d, J=8.82Hz, 2H, ArH), 7.85 (ddd, J=8.32,5.64,2.51Hz, 1H, ArH), 7.67 (d, J=8.51Hz, 2H, ArH), 7.36 (d, J=8.80Hz, 2H, ArH), 7.08 (d, J=7.27Hz, 2H, ArH), 7.03 (d, J=6.81Hz, 1H, ArH); LCMS (APCI ⁺): 431 (50%), 433 (100%); HPLC:100%.

Embodiment NN.

(E)-N-(4-(1-((diamino methene base) hydrazono-) ethyl) phenyl)-4-(6-(dimethylamino) quinolyl-4 amino) benzamide dihydrochloride (compound N N) synthetic

(i) 6-(dimethylamino)-4-chloroquinoline/1:2 MeOH:H ₂O, c.HCl refluxes 32 hours

(E)-N-(4-(1-((diamino methene base) hydrazono-) ethyl) phenyl)-4-(6-(dimethylamino) quinolyl-4 amino) benzamide dihydrochloride (compound N N).(0.22g is 0.56mmol) at 1:2 EtOH:H to amine J1 ₂(0.13g is 0.61mmol) at 1:2 EtOH:H sequentially to add 6-(dimethylamino)-4-chloroquinoline among the O (20mL) ₂Solution among the O (10mL) and dense HCl (0.17mL, 5.5mmol).The mixture that obtains was refluxed several hours, make its ambient temperature overnight of rising again then.(16 hours) afterwards, TLC analyzes and (uses N-BuOH:H ₂O:CH ₃CO ₂Therefore the last phase wash-out of the 5:4:1 mixture of H) be presented at some 6 residues are arranged in the reaction mixture, (0.22g 0.56mmol), and refluxes other mixture 16 hours to add another normal J1.After this, TLC analyzes and shows that quinoline almost completely consumes, and therefore the solvent decompression is removed, and resistates is passed through three MeOH azeotropic cyclic dryings.Resistates is from MeOH (with the hcl acidifying 1.25 methyl alcohol): the EtOAc redeposition, and by preparation property HPLC through single step purification, obtain compound N N, be unbodied yellow-brown solid (18mg, 6%); Mp (MeOH:EtOAc)〉280 ℃; ¹H NMR[(CD ₃) ₂SO]: δ 2.33[s, 3H, ArC (CH ₃)=N-], 3.11[s, 6H, ArN (CH ₃) ₂], 7.00 (s, 1H, ArH], 7.41[s, 1H, ArH], 7.57-7.90[m, 8H, ArH ﹠amp;=C (NH ₂) ₂], 7.99[d, J=8.45Hz, 2H, ArH], 8.12[d, J=7.01Hz, 2H, ArH], 8.35[br s, 1H, ArH] and, 9.86[br s, 1H, ArH], 10.40[s, 1H, ArH] and, 10.86[br s, 1H, ArNHAr] { its excess-three tradable H signal is invisible }; LCMS (APCI ⁺): 482 (100%); HPLC:98.7%.

Embodiment OO.

6-(dimethylamino)-4-[4-({ 3-[(1-methyl-4-pyridyl) amino] benzoyl } amino) anilino]-quinoline dichloride (compound 001) and related compound (compound 002) synthetic

(i) 6-(dimethylamino)-4-chloroquinoline/EtOH/H ₂O (2:1)/c.HCl/ refluxes

6-(dimethylamino)-4-[4-(3-[(1-methyl-4-pyridyl) and amino] benzoyl } amino) anilino]-quinoline dichloride Sg EPG 133 (compound 001).To amine K1 (151mg, 0.43mmol) solution in ethanol (28mL) and water (14mL) be added in 6-(dimethylamino)-4-chloroquinoline in the ethanol (5mL) (107mg, 0.52mmol) and two dense HCl.With reaction mixture refluxed 3 days,, make its boiling and make it be cooled to 20 ℃ with EtOAc (150mL) dilution.The throw out that obtains is filtered,, obtain yellow solid 194mg, it is passed through preparation property HPLC purifying (TFA/CH with more EtOAc washing and dry ₃CN), obtain (3) (80mg, 33%) then from the MeOH/EtOAC recrystallization; 17-173 ℃ of mp (MeOH/EtOAc) (dec); ¹H NMR[(CD ₃) ₃SO] δ 13.88 (bs, 1H, N ⁺H), 10.64 (s, 1H, NH), 10.53 (s, 1H, NH), 10.25 (bs, 1H, NH), 8.32 (d, J=d, J=7.5Hz, 2H, ArH), 8.27 (d, J=6.5Hz, 1H, ArH), 7.98 (d, J=8.9Hz, 2H, ArH), 7.94 (d, J=8.0Hz, 1H, ArH), 7.90 (bs, 1H, ArH), 7.84 (d, J=9.4Hz, 1H, ArH), 7.69 (t, J=7.9Hz, 1H, ArH), 7.64 (dd, J=9.4,2.6Hz, 1H, ArH), 7.58 (dd, J=8.0,1.3Hz, 1H, ArH), 7.48-7.45 (m, 3H, ArH), 7.22 (d, J=7.5Hz, 2H, ArH), 6.68 (s, 1H, ArH), 3.99 (s, 3H, N ⁺CH ₃), 3.12[s, 6H, N (CH ₃) ₂]; APCI ve ⁺489.

6-(dimethylamino)-4-[4-(4-[(1-methyl-4-pyridyl) and amino] benzoyl } amino)-anilino] quinoline dichloride SG EPG 134 (compound 002).To amine K2 (162mg, 0.46mmol) solution in ethanol (28mL) and water (14mL) be added in 6-(dimethylamino)-4-chloroquinoline in the ethanol (5mL) (114mg, 0.55mmol) and two dense HCl.With reaction mixture refluxed 2 days,, make its boiling and make it be cooled to 20 ℃ with EtOAc (150mL) dilution.The throw out that obtains is filtered,, obtain yellow solid 161mg, it is passed through preparation property HPLC purifying (TFA/CH with more EtOAc washing and dry ₃CN), then from the MeOH/EtOAC recrystallization, obtain (5) (80mg, 31%); 156 ℃ of mp (MeOH/EtOAc) (dec); ¹H NMR[(CD ₃) ₂SO] δ 13.90 (bs, 1H, N ⁺H), 10.73 (s, 1H, NH), 10.48 (s, 1H, NH), 10.29 (bs, 1H, NH), 8.36 (d, J=7.5Hz, 2H, ArH), 8.28 (d, J=6.7Hz, 1H, ArH), 8.11 (d, J=8.6Hz, 2H, ArH), 8.00 (d, J=8.8Hz, 2H, ArH), 7.84 (d, J=9.4Hz, 1H, ArH), 7.65 (dd, J=9.4,2.2Hz, 1H, ArH), 7.52-7.45 (m, 5H, ArH), 7.28 (d, J=7.5Hz, 2H, ArH), 6.68 (d, J=6.8Hz, 1H, ArH), 4.01 (s, 3H, N ⁺CH ₃), 3.12[s, 6H, N (CH ₃) ₂] APCI ve ⁺489.

Embodiment PP.

4-[(1E)-N-(diamino methene base) ethane hydrazono-]-preparation of N-(4-{[6-(dimethylamino)-4-quinolyl] amino } phenyl) benzamide dihydrochloride (Compound P P)

(i) c.HCl/MeOH/ backflow/H ₂NN=C (NH ₂) ₂(ii) EDCI/DMAP/DMF/RT

4-[(1E)-and N-(diamino methene base) ethane hydrazono-] benzoate hydrochlorate (L2).(1.041g, 6.34mmol), (1.3eq) (0.7ml's aminoguanidin carbonate 7.0mmol) refluxed 1 hour at MeOH (30mL) with dense HCl for 1.12g, 8.2mmol with 4-acetylbenzoic acid (L1).Reaction mixture dilutes with EtOAc, be cooled to 20 ℃ and the throw out that obtains filtered and, obtain L2 (887mg, 55%), mp (MeOH/EtOAc) from the MeOH/EtOAc recrystallization 300 ℃; ¹H NMR ([(CD ₃) ₂SO] and δ 7.97-7.89 (m, 4H, ArH), 6.75 (br, 4H, 2 * NH ₂), 2.25 (s, 3H, CH3), mass spectrum APCI ⁺221.

4-[(1E)-N-(diamino methene base) ethane hydrazono-]-N-(4-{[6-(dimethylamino)-4-quinolyl] amino } phenyl) benzamide dihydrochloride (Compound P P).Will be at the N among the DMF (10mL) ⁴-(4-aminophenyl)-N ⁶, N ⁶-dimethyl-4,6-quinoline diamines (A6) (108mg, 0.34mmol), L2 (107mg, 0.34mmol), EDCI (160mg, 0.68mmol) and DMAP (101mg 0.68mmol) stirred 72 hours at 20 ℃.With solvent at 55 ℃ of reduction vaporizations.Residue diluted with water is also used NH ₃Aqueous solution alkalization.The throw out that obtains is filtered, wash dry air and chromatogram purification (SiO with water ₂/ DCM/MeOH/aq NH ₃0-7% 2% NH ₃).To comprise the fraction merging of correct quality and be evaporated to drying, obtain the 120mg yellow solid.The following HCl of being translated into salt: contain 1 of 4N HCl to several of the suspension in MeOH, the 4-diox arrives solvent evaporation dry then.The resistates that obtains obtains crude product (107mg) from the MeOH/EtOAc recrystallization, analyzes demonstration by HPLC and comprises two main compound.It is passed through preparation property HPLC purifying (HCOO ^-N ⁺H ₄), obtain Compound P P (52mg, 27%); M.p (MeOH/EtOAc)〉300 ℃; ¹H NMR[(CD ₃) ₂SO] δ 14.09 (brs, 1H, N ⁺H), 11.21 (s, 1H, NH), 10.56 (s, 1H, NH), 10.40 (s, 1H, NH), 8.27 (d, J=6.8Hz, 1H, ArH), 8.13 (d, J=8.6Hz, 2H, ArH), 8.06-8.01 (m, 4H, ArH), 7.88 (d, J=9.4Hz, 1H, ArH), 7.83 (br, 4H, 2 * NH ₂), 7.65 (dd, J=9.4,2.3Hz, 1H, ArH), 7.53 (d, J=2.3Hz, 1H, ArH), 7.47 (d, J=8.9Hz, 2H, ArH), 6.67 (d, J=6.8Hz, 1H, ArH), 3.13[s, 6H, [N (CH ₃) ₂], 2.41 (s, 3H, CH ₃); Mass spectrum APCI ⁺481.

Embodiment QQ.

4-[4-({ 3-[(1E)-N-(diamino methene base) ethane hydrazono-]-benzoyl } amino) anilino]-preparation of 6-(dimethylamino) quinoline muriate (compound Q Q)

(i) EDCI/DMAP/DMF/20 ℃; (ii) MeOH/HCl/ refluxes

4-{4-[(3-acetylbenzene formyl radical) amino] anilino }-6-(dimethylamino) quinoline muriate (M2).With A3 (181mg, 0.58mmol), 3-acetylbenzoic acid (M1) (97mg, 0.58mmol) and EDCI (220mg, 0.1.16mmol) mixture in DMF (5mL) stirred 5 minutes at 20 ℃.(140mg 1.16mmol) and with reaction mixture stirred 24 hours at 20 ℃ to add DMAP then.Solvent decompression removed and with resistates at NaHCO ₃Stirred 1 hour in the aqueous solution.The throw out that obtains filtered and pass through SiO ₂In chromatography purification, the gradient elution with the MeOH/DCM of (0-7.5%) obtains M2 (113mg, 42%); Mp (DCM/MeOH)〉280 ℃; ¹H NMR[(CD ₃) ₂SO] δ 14.02 (br, 1H, N ⁺H), 10.66 (s, 1H, NH), 10.31 (s, 1H, NH), 8.53 (t, J=1.6Hz, 1H, ArH), 8.28 (d, J=6.7Hz, 1H, ArH), 8.24 (td, J=8.1,1.5Hz, 1H, ArH), 8.19 (td, J=7.8,2.8Hz, 1H, ArH), 8.00 (d, J=6.8Hz, 2H, ArH), 7.87 (d, J=9.4Hz, 1H, ArH), 7.78 (t, J=7.8Hz, 1H, ArH), 7.64 (dd, J=9.4,2.6Hz, 1H, ArH), 7.51 (d, J=2.5Hz, 1H, ArH), 7.47 (d, J=8.8Hz, 2H, ArH), 6.69 (d, J=6.7Hz, 1H, ArH), 3.09[s, 6H, (NCH ₃) ₂], 2.68 (s, 3H, COCH ₃); APCI ⁺Ve 425.

4-[4-(3-[(1E)-N-(diamino methene base) ethane hydrazono-] benzoyl } amino) anilino]-6-(dimethylamino) quinoline muriate (compound Q Q).With M2 (94mg, 0.20mmol), aminoguanidin carbonate (42mg, 0.3mmol) and dense HCl (0.02mL, 0.022mmol) mixture in MeOH (10mL) refluxed 2 hours, with the EtOAc dilution, some MeOH are seethed with excitement and is cooled to 20 ℃.The throw out that obtains is filtered,, obtain compound Q Q (109mg 100%), be yellow solid with more EtOAc washing and from the MeOH/EtOAc recrystallization; Mp (MeOH/EtOAC)〉280 ℃; ¹H NMR[(CD ₃) ₂SO] δ 14.13 (br1H, N ⁺H), 11.25 (s, 1H, NH), 10.67 (s, 1H, NH), 10.42 (s, 1H, NH), 8.48 (t, J=1.5Hz, 1H, ArH), 8.27 (d, J=6.8Hz, 1H, ArH), 8.22 (t, d, J=7.9,1.4Hz, 1H, ArH), 8.05-8.01 (m, 3H, ArH), 7.89 (d, J=9.6Hz, 1H, ArH), 7.85 (br, 4H, 2 * NH ₂), 7.66-7.59 (m, 2H, ArH), 7.54 (d, J=2.3Hz, 1H, ArH), 7.47 (d, J=8.8Hz, 2H, ArH), 6.67 (d, J=6.8Hz, 1H, ArH), 3.13[s, 3,6H, (NCH ₃) ₂], 2.45 (s, 3H, CH ₃); APCI ⁺Ve 481.

Embodiment RR.

3-[(2,6-diamino-4-pyrimidyl) amino]-N-{4-[(6-nitro-4-quinolyl) amino] phenyl } preparation of benzamide dihydrochloride (compound R R1) and related compound (compound R R2 and compound R R3)

(i) 4-chloroquinoline/EtOH/H ₂O/h+/backflow; (ii) H ₂/ Pd/C/MeOH

3-[(2,6-diamino-4-pyrimidyl) amino]-N-{4-[(6-nitro-4-quinolyl) amino] phenyl } benzamide dihydrochloride (compound R R1).(204mg is 0.55mmol) at EtOH (20mL) and H to amine B6 ₂Solution among the O (10mL) adds 4-chloro-6-nitroquinoline, and (126mg 0.61mmol) and stir up to its dissolving, adds 2 dense HCl then.With reaction mixture refluxed 4 hours,, make its boiling and be cooled to 20 ℃ with the EtOAc dilution.The throw out that obtains is filtered and, obtain compound R R1 (224mg 70%) from the MeOH/EtOAc recrystallization; M.p. (MeOH/EtOAc)〉300 ℃; ¹H NMR[(CD ₃) ₂SO] δ 14.75 (br, 1H, N ⁺H), 11.50 (br, 1H, N ⁺H), 11.10 (br, 1H, NH), 10.50 (s, 1H, NH), 9.88 (s, 1H, NH), 9.77 (s, 1H, NH), 9.76 (s, 1H, ArH), 8.66 (d, J=9.3Hz, 1H, ArH), 8.60 (d, J=6.8Hz, 1H, ArH), 8.19 (d, J=9.3Hz, 1H, ArH), 8.25 (d, J=8.8Hz, 2H, ArH), 7.94 (br s 2H, NH ₂), 7.71 (d, J=7.6Hz, 1H, ArH), 7.56-7.42 (m, 6H, NH ₂﹠amp; 4 * ArH), 6.91 (d, J=6.8Hz, 1H, ArH), 5.44 (s, 1H, ArH).HRMS (FAB ⁺) calculated value C ₂₆H ₂₂N ₉O ₃(M ⁺1) m/z 508.1846, measured value 508.1841.

3-[(2-amino-6-methyl-4-pyrimidyl) amino]-N-{4-[(6-nitro-4-quinolyl) amino] phenyl } benzamide dihydrochloride (compound R R2).(205mg is 0.55mmol) at EtOH (20mL) and H to amine B3 ₂Solution among the O (10mL) adds 4-chloro-6-nitroquinoline, and (135mg 0.64mmol) and stir up to dissolving, adds 2 dense HCl then.With reaction mixture refluxed 5 hours,, make its boiling and be cooled to 20 ℃ with the EtOAc dilution.The throw out that obtains is filtered and, obtain compound R R2 (217mg 68%) from the MeOH/EtOAc recrystallization; M.p. (MeOH/EtOAc)〉300 ℃; ¹H NMR[(CD ₃) ₂SO] δ 13.00 (br, 2H, 2 * N ⁺H), 10.99 (br, 1H, NH), 10.75 (br s, 1H, NH), 10.53 (s, 1H, NH), 9.75 (d, J=1.9Hz, 1H, ArH), 8.64 (br d, J=7.4Hz, 1H, ArH), 8.60 (d, J=6.7Hz, 1H, ArH), 8.17 (d, J=9.3Hz, 1H, ArH), 8.15 (br, 1H, NH), 8.07 (br s, 1H, NH), 7.99 (d, J=8.8Hz, 2H, ArH), 7.79 (br d, J=7.5Hz, 3H, ArH), 7.57 (t, J=7.9Hz, 1H, ArH), 7.48 (d, J=8.8Hz, 2H, ArH), 6.91 (d, J=6.7Hz, 1H, ArH), 6.22 (d, J=0.6Hz, 1H, ArH), 2.30 (s, 3 hours.CH ₃); HRMS (FAB ⁺) calculated value C ₂₇H ₂₃N ₈O ₃(M ^{+ 1}) m/z 507.1893, measured value 507.1888.

3-[(2-amino-6-methyl-4-pyrimidyl) amino]-N-{4-[(6-amino-4-quinolyl base) amino] phenyl } benzamide dihydrochloride (compound R R3).With compound R R2 (146mg, 146mg, 0.25mmol) be dissolved in MeOH (30ml) and with 10% Pd/C (20mg) 30Hgmm hydrogenation 3 hours.With reaction mixture filtration passing through Celite pad.Filtrate is evaporated to drying.The resistates that obtains is dissolved in MeOH (10mL), stirs, precipitate by adding EtOAc then, filter and drying, obtain 120mg compound 7 with the MeOH that contains 1.25M HCl (0.5mL).It is analyzed by HPLC is 90% purity.Pass through at NH then ₃Stir in the aqueous solution and be translated into free alkali, filter, dry air and on neutral alumina chromatogram purification, with comprising 1% NH ₃The gradient elution of (0-10%) MeOH/DCM of the aqueous solution obtains the pure free alkali of 83mg.Be translated into HCl salt by it being dissolved in the MeOH that MeOH and adding contain 1.25M HCl then.Be evaporated to drying and with resistates from the MeOH/EtOAc recrystallization, obtain compound R R3 (88mg, 64%), 280-284 ℃ of mp (MeOH/EtOAc); ¹H NMR[(CD ₃) ₂SO] δ 14.0 (d, J=5.4Hz, 1H, N ⁺H), 12.78 (br s, 1H, N ⁺H), 10.79 (br s, 1H, NH), 10.50 (s, 1H, NH), 10.19 (s, 1H, NH), 8.22 (t, J=6.4Hz, 2H, ArH), 8.16-8.07 (br, 2H, NH ₂) 7.96 (d, J=8.9Hz, 2H, ArH), 7.80 (br s, 1H, ArH), 7.78 (d, J=9.0Hz, 2H, ArH), 7.56 (t, J=8.0Hz, 1H, ArH), 7.47 (br d, J=2.1Hz, 1H, ArH), 7.43 (d, J=8.7Hz, 2H, ArH), 7.39 (dd, J=9.0,2.2Hz, 1H, ArH), 6.67 (d, J=6.8Hz, 1H, ArH), 6.23 (s, 1H, ArH), 5.9 (v.br, 2H, NH ₂), 2.31 (s, 3H, CH ₃).

Embodiment SS.

4-[(2,6-diamino-4-pyrimidyl) amino]-preparation of N-(4-{[6-(dimethylamino)-4-quinolyl] amino } phenyl) benzamide dihydrochloride (compound S S)

(i) ethanol/H ⁺/ reflux; (ii) A3/EDCI/DMAP/N-methyl-2-pyrrolidone

4-[(2,6-diamino-4-pyrimidyl) amino] phenylformic acid (01).(2.0g, 14.55mmol) and 2, (2.013g 14.55mmol) is dissolved in cellosolvo (20mL) to 6-diamino-4-chloropyrimide (B4) with 4-benzaminic acid (D1).Add 2 dense HCl and refluxed 20 hours to this mixture.Reaction mixture is cooled to 20 ℃ and the throw out that obtains filtered and from the MeOH/EtOAc recrystallization, obtain compound 01 (3.12g, 56%), mp (MeOH/EtOAc) ℃; ¹H NMR[(CD ₃) ₂SO] δ 12.65 (br, 1H, COOH or N ⁺H), 11.84 (br s, 1H, N ⁺H or COOH), 10.07 (s, 1H, NH), 7.86 (brd, J=8.7Hz, 2H, ArH), 7.75 (v.br d, J=7.8,2H, ArH), 7.64 (br, 2H, NH ₂), 7.51 (br, 2H, NH ₂), 5.50 (s, 1H, ArH).

4-[(2,6-diamino-4-pyrimidyl) amino]-N-(4-{[6-(dimethylamino)-4-quinolyl] amino } phenyl) benzamide dihydrochloride (compound S S).Will the compound 01 in the N-Methyl pyrrolidone (5mL) (101.4mg, 0.36mmol), EDCI (138mg, 0.72mmol) and DMAP (88mg 0.36mmol) stirred 5 minutes at 20 ℃.Add then compound 11 (100mg, 0.36mmol) and Et ₃N (0.2mL, 1.44mmol) and stirred 20 hours.TLC (the Al of small sample ₂O ₃/ DCM/MeOH 5% and/NH ₃The aqueous solution) show and to still have compound 11 to exist, therefore add more EDCI (138mg, 0.72mmol) and stirred 72 hours.Reaction mixture H then ₂O dilutes and stirred 1 hour.The throw out that obtains is filtered, washes with water, dry air and on neutral alumina chromatogram purification, with the gradient elution of of 0-5%DCM/MeOH,, add 1% NH then to remove unreacted 11 impurity ₃The aqueous solution is with eluted product 12.To comprise the fraction evaporation of product, obtain compound 12 (75mg).It is dissolved in a spot of MeOH and stir with the MeOH that contains 1.25M HCl (0.5mL), with solvent evaporation and with resistates from the MeOH/ recrystallization, obtain compound S S (84mg, 40%), 283-287 ℃ of mp (MeOH/EtOAc); ¹H NMR[(CD ₃) ₂SO] δ 14.14 (d, J=4.7Hz .1H, N ⁺H), 11.81 (br, 1H, N ⁺H), 10.42 (s, 1H, NH), 10.39 (s, 1H, NH), 10.09 (s, 1H, NH), 8.26 (t, J=6.4Hz1H, ArH), 8.02-7.98 (m, 4H, ArH), 7.89 (d, J=8.7Hz, 1H, ArH), 7.80 (br d, 2H, ArH), 7.68 (br, 2H, NH ₂), 7.64 (dd, J=9.4,2.5Hz, 1H, ArH), 7.53 (dd, J=9.3,2.5Hz, 1H, ArH), 7.52 (br s, 2H, NH ₂), 7.45 (d, J=8.9Hz, 2H, ArH), 6.66 (d, J=6.8Hz, 1H, ArH), 5.51 (s, 1H, ArH), 3.12[s, 6H, N (CH ₃) ₃].

Embodiment TT.

4-[(2,6-diamino-4-pyrimidyl) amino]-N-[4-(4-quinolyl amino) phenyl] preparation of benzamide dihydrochloride (compound TT)

(i) MeOH/HCl/ refluxes; (ii) H ₂/ Pd/C/MeOH; (iii) 4-chloroquinoline/2:1 EtOH/H ₂O/H ⁺Reflux

4-[(2,6-diamino-4-pyrimidyl) amino]-N-(4-nitrophenyl) benzamide hydrochloride salt (P2).By heating with amine P1 (1.0g, 3.89mmol) and chloropyrimide B4 (1.12g 7.78mmol) is dissolved in MeOH (200mL), adds dense HCl (3) then and refluxes 5 days.Reaction mixture is cooled to 20 ℃ and throw out filtered,, obtains pure basically Compound P 2 (814mg 52%) with more MeOH washing and drying, ¹H NMR[(CD ₃) ₂SO] δ 11.84 (s, 1H, N ⁺H), 10.75 (s, 1H, NH), 10.12 (s, 1H, NH), 8.28-8.24 (m, 2H, ArH), 8.11-8.07 (m, 2H, ArH), 8.24 (br d, J=8.7Hz, 2H, ArH), 7.83 (br, 2H, ArH), 7.68 (br, 2H, NH ₂), 7.54 (br, 2H, NH ₂), 5.51 (s, 1H, ArH).HRMS (FAB ⁺) calculated value C ₁₅H ₁₆N ₇O ₃(M ^{+ 1}) m/z366.1315, measured value 366.1306.

N-(4-aminophenyl)-4-[(2,6-diamino-4-pyrimidyl) amino] benzamide dihydrochloride (P3).With Compound P 2 (811mg, 2.01mmol) suspension in MeOH100 (mL) with 10% Pd/C (100mg) at 40Hg mm H ₂Hydrogenation is 20 hours under the pressure.The new suspension that obtains is stirred with the MeOH that contains 1.25M HCl (5mL) so that lysate filters by Celite pad it to remove the Pd resistates then.Filtrate is evaporated to drying and with resistates from the MeOH/EtOAc recrystallization, obtain Compound P 3 (785mg, 100%), ¹H NMR[(CD ₃) ₂SO] δ 10.02 (s, 1H, NH), 9.88 (s, 1H, NH), 7.91 (d, J=8.7Hz, 2H, ArH), 7.74 (d, J=8.5Hz, 2H, ArH), 7.62 (br, 2H, NH ₂), 7.68 (br s, 2H, NH ₂), 7.47 (d, J=8.8Hz, 2H, ArH), 6.71 (d, J=8.7Hz, 2H, ArH), 5.45 (s, 1H, ArH).

4-[(2,6-diamino-4-pyrimidyl) amino]-N-[4-(4-quinolyl amino) phenyl] benzamide dihydrochloride (compound TT).(128mg is 0.34mmol) at EtOH (20mL) and H to Compound P 3 ₂Solution among the O (10mL) adds the 4-chloroquinoline, and (127mg 0.51mmol) and stir up to its dissolving, adds 2 dense HCl then.With reaction mixture refluxed 4 hours,, make its boiling and be cooled to 20 ℃ with the EtOAc dilution.The throw out that obtains filters and from the MeOH/EtOAc recrystallization, obtains compound TT (151mg, 83%); M.p. (MeOH/EtOAc) 263-267 ℃; ¹H NMR[(CD ₃) ₂SO] δ 14.10 (br, 1H, N ⁺H), 11.90 (br, 1H, N ⁺H), 10.89 (s, 1H, NH), 10.40 (s, 1H, NH), 10.04 (s, 1H, NH), 8.77 (d, J=8.7Hz, 1H, ArH), 8.51 (d, J=7.0Hz, 1H, ArH), 8.06-7.98 (m, 6H, ArH), 7.83-7.79 (m, 3H, ArH﹠amp; NH ₂), 7.61 (br, 2H, NH ₂), 7.47 (d, J=8.9Hz, m, 2H, ArH), 6.78 (d, J=7.0Hz, 1H, ArH), 5.45 (s, 1H, ArH).

Embodiment UU.

N-{4-[(6-amino-4-quinolyl base) amino] phenyl }-3-[(2,6-diamino-4-pyrimidyl) amino] preparation of benzamide dihydrochloride (compound UU)

(i)H ₂/Pd/C/MeOH

N-{4-[(6-amino-4-quinolyl base) amino] phenyl }-3-[(2,6-diamino-4-pyrimidyl) amino] benzamide dihydrochloride (compound UU).With 3-[(2,6-diamino-4-pyrimidyl) amino]-N-{4-[(6-nitro-4-quinolyl) amino] phenyl benzamide dihydrochloride (compound R R1) (148mg, 0.25mmol) be dissolved in MeOH (30ml) and with 10%Pd/C (20mg) hydrogenation 20 hours under 30Hg mm.With reaction mixture filtration passing through Celite pad.Filtrate is evaporated to drying.The resistates that obtains is dissolved in MeOH (10mL), stirs, precipitate by adding EtOAc then, filter and drying, obtain 90mg compound 1 with the MeOH that contains 1.25M HCl (0.5mL).It is analyzed by HPLC is 78% purity.By being stirred, it is translated into free alkali then in the NH3 aqueous solution, filter, dry air and on neutral alumina chromatogram purification, with (0-7%) wash-out of the MeOH/DCM that comprises the 1.5% NH3 aqueous solution, obtain the pure free alkali (51mg, 43%) of the SN 31043 of 51mg.Be translated into HCl salt by it being dissolved in the MeOH that MeOH and adding contain 1.25M HCl then.Be evaporated to drying and with resistates from the MeOH/EtOAc recrystallization, obtain compound UU, 250-255 ℃ of mp (MeOH/EtOAc); ¹H NMR[(CD ₃) ₂SO] δ 14.15 (d, J=6.1Hz, 1H, N ⁺H), 11.70 (br 1H, N ⁺H), 10.48 (s, 1H, NH), 10.46 (s, 1H, NH, 10.21 (S, 1H, NH), 8.22 (t, J=6.5Hz, 1H, ArH), 8.05-7.89 (m, 4H, ArH), 7.79 (d, J=9.1Hz, 1H, ArH), 7.71 (brd, J=7.5Hz, 1H, ArH), 7.59 (br, 2H, NH ₂), 7.53-7.38 (m, 8H, ArH ﹠amp; NH ₂), 6.67 (d, J=6.7Hz, 1H, ArH), 5.45 (s, 1H, ArH).HPLC purity %.HRMS (FAB ⁺), calculated value C ₂₆H ₂₃N ₉O (M ^{+ 1}) m/z 478.2104, measured value, 478.2103.

EXAMPLE V V.

(2,6-diamino-4-pyrimidyl) amino]-N-{4-[(6-nitro-4-quinolyl) amino] phenyl } preparation of benzamide dihydrochloride (compound VV1) and related compound (compound VV2)

4-[(2,6-diamino-4-pyrimidyl) amino]-N-{4-[(6-nitro-4-quinolyl) amino] phenyl } benzamide dihydrochloride (compound VV1).(288mg is 0.76mmol) at EtOH (20mL) and H to Compound P 3 ₂Solution among the O (10mL) adds 4-chloro-6-nitroquinoline, and (196mg 0.94mmol) and stir up to its dissolving, adds 2 dense HCl then.With reaction mixture refluxed 4 hours,, make its boiling and be cooled to 20 ℃ with the EtOAc dilution.The throw out that obtains is filtered and, obtain compound VV1 (445mg99%) from the MeOH/EtOAc recrystallization; M.p. (MeOH/EtOAc) 260-265 ℃; ¹H NMR[(CD ₃) ₂SO] δ 11.38 (br s, 1H, N ⁺H), 10.43 (s, 1H, NH), 10.10 (s, 1H, NH), 9.81 (d, J=2.1Hz, 1H, NH), 8.78 (dd, J=9.3,2.2Hz, 1H, ArH), 8.84 (d, J=7.2Hz, 1H, ArH), 8.24 (d, J=9.3Hz, 1H, NH), 8.04-7.98 (m, 4H, ArH), 7.81 (br d, J=7.4Hz, 2H, ArH), 7.68 (brs, 2H, NH ₂), 7.53 (br s, 2H, NH ₂), 7.48 (d, J=8.9Hz, 2H, ArH), 6.90 (d, J=7.0Hz, 1H, ArH), 5.52 (s, 1H, ArH).HPLC purity 100%; HRMS (FAB ⁺) calculated value C ₂₆H ₂₁N ₉O ₃(M ^{+ 1}) m/z 508.1846, measured value 508.1844; Ultimate analysis calculated value C ₂₆H ₂₃Cl ₂N ₉O ₃.4H ₂O:C, 47.9; H, 4.2; N, 19.3; Cl, 10.9; Measured value C, 48.0; H, 4.4; N, 19.2; Cl, 11.1%.

N-{4-[(6-amino-4-quinolyl base) amino] phenyl }-4-[(2,6-diamino-4-pyrimidyl) amino] benzamide dihydrochloride (compound VV2).With compound VV1 (211mg, 0.36mmol) be dissolved in MeOH (30ml) and with 10% Pd/C (20mg) 30Hgmm hydrogenation 5 hours.With reaction mixture filtration passing through Celite pad.Filtrate is evaporated to drying.The resistates that obtains is dissolved in MeOH (10mL), stirs with the MeOH that contains 1.25M HCl (0.5mL), then MeOH is evaporated to drying.Resistates is dissolved in MeOH again and is evaporated to drying,, obtain the 178mg product then from the MeOH/EtOAc recrystallization; Its by HPLC analyze have only 93% pure.The following then free alkali that is translated into: at NH ₃Stir in the aqueous solution, filter, dry air and on neutral alumina chromatogram purification, with comprising 1.5%NH ₃The MeOH/DCM gradient elution of (0-7.5%) of the aqueous solution obtains the free alkali of pure compound VV2.The following then HCl of being translated into salt: it is dissolved in the MeOH that MeOH and adding contain 1.25MHCl.It is evaporated to drying and with resistates from the MeOH/EtOAc recrystallization, obtain compound VV2 (138mg, 70%), 262-266 ℃ of mp (MeOH/EtOAc); ¹H NMR[(CD ₃) ₂SO] δ 14.11 (br s, 1H, N ⁺H), 11.81 (br, 1H, N ⁺H), 10.36 (s, 1H, NH), 10.19 (s, 1H, NH), 10.08 (s, 1H, NH), 8.21 (t, J=6.0Hz, 1H, ArH), 8.00-7.96 (m, 4H, ArH), 7.79 (br s, 2H, NH ₂), 7.78 (d, J=9.1Hz, 1H, ArH), 7.67 (br s, 2H, NH ₂), 7.53 (br s, 2H, NH ₂), 7.48 (br s, 1H, ArH), 7.42-7.38 (m, 3H, ArH), 6.66 (d, J=6.7Hz, 1H, ArH), 6.00 (v.br, 2H, NH ₂), 5.51 (s, 1H, ArH).HPLC purity 99.7%; HRMS (FAB ⁺) calculated value C ₂₆H ₂₄N ₉O (M ⁺1) m/z 478.2104 measured values 478.2107.

Embodiment WW.

N-methyl-N-[4-(pyridin-4-yl amino) phenyl]-preparation of 4-(quinolyl-4 amino) benzamide hydrochloride salt (compound WW)

N-methyl-4-nitro-N-[4-(pyridin-4-yl amino) phenyl] benzamide hydrochloride salt (3).To amine 1 (0.82g, no Shui diox (70mL) solution 4.10mmol) sequentially add anhydrous pyridine (1.65mL, 20.50mmol) and acyl chlorides 2 (2.08g 11.19mmol), and refluxes the mixture that obtains～60 hours.After this, with the reaction mixture cool to room temperature, and by filtering the solid that collection obtains.By adding NH ₃The aqueous solution alkalizes filtrate, and collects the second batch of solid that obtains by filtering.The solid batch of material is merged, obtain acid amides 3,, it is passed through for unbodied yellow solid (3.46g) ¹H NMR and MS analyze, and without using through single step purification. ¹H NMR:8.88 (dd, J=6.32,1.32Hz, 4H, ArC (O) N (CH ₃) Ar ﹠amp; ArNHAr), 8.32 (ddd, J ,=9.17,4.31,2.27Hz, 4H, ArH), 8.17 (ddd, J=9.17,4.31,2.28), 7.98 (dd, J=7.59,6.50Hz, 4H, ArH) [pyridyl-N ⁺-H is invisible]; LCMS (APCI ⁺): 349 (100%).

4-amino-N-methyl-N-[4-(pyridin-4-yl amino) phenyl] benzamide hydrochloride salt (4).To acid amides 3 (3.46g, MeOH 8.99mmol) (～40mL) solution adds 10% Pd/C of a spatula head, and with the suspension that obtains 40psi hydrogenation 16 hours.After this, reaction mixture is filtered by Celite pad, and the solvent decompression is removed.Resistates obtains amine 4 from the MeOH-EtOAc redeposition, is unbodied cream-colored solid (0.25g, 15%from l); Mp 235-238 ℃ (powder-tarry materials), 245-249 ℃ (emitting gas); ¹HNMR:13.85 (v br s, 1H, pyridine-N ⁺-H), 10.75 (s, 1H, ArNHAr), 8.28 (d, J=7.23Hz, 2H, ArH), 7.25 (s, 4H, ArH), 7.16 (d, J=8.44Hz, 2H, ArH), 7.07 (d, J=7.03Hz, 2H, ArH), 6.73 (d, J=7.79Hz, 2H, ArH), 3.36 (s, ArC (O) N (CH ₃) Ar) [ArN ⁺H ₃Invisible]; HRMS (EI) calculated value C ₁₉H ₁₈N ₄Om/z 318.1481, measured value 318.1481.

N-methyl-N-[4-(pyridin-4-yl amino) phenyl]-4-(quinolyl-4 amino) benzamide hydrochloride salt compound WW. is to amine 4 (0.23g, 0.58mmol) solution in the 20% EtOH aqueous solution (40mL) sequentially adds quinoline 5 (0.22g, 1.36mmol) and dense HCl (0.17mL, 5.60mmol), and the mixture that obtains refluxed～16 hours.After this, solvent decompression is removed, and with resistates from MeOH: the EtOAc redeposition, obtain compound WW, be light yellow amorphous solid (0.27g, 91%), mp 287-292 ℃ (tarry materials-liquid); ¹H NMR:14.6 (v br s, 1H, quinolyl-N ⁺-H), 13.84 (v br s, 1H, pyridyl-N ⁺-H, 1H), 11.02 (s, 1H, Ar NHAr), 10.95 (s, 1H, ArNHAr), 8.80 (d, J=8.49Hz, 1H, ArH), 8.58 (d, J=6.95Hz, 1H, ArH), 8.28 (d, J=7.28Hz, 2H, ArH), 8.06 (m, 2H, ArH), 7.79 (ddd, J=8.31,6.80,1.30Hz, 1H, ArH), 7.49 (d, J=8.50Hz, 1H, ArH), 7.40 (d, J=8.53Hz, 1H, ArH), 7.36 (d, J=8.78Hz, 1H, ArH), 7.31 (m, 4H), 7.13 (d, J=7.10Hz, 2H, ArH), 6.76 (d, J=6.94Hz, 1H, ArH), 3.44 (s, 3H, ArC (O) N (CH ₃) Ar); HRMS (FAB ⁺): calculated value C ₂₈H ₂₄N ₅O (MH ⁺) m/z 446.1981, measured value 446.1985; HPLC:96.3%.

Embodiment XX.

N-{4-[(2-amino-6-methyl-4-pyrimidyl) amino] phenyl }-preparation of 5-(4-quinolyl amino)-2-pyridine carboxamide dihydrochloride (compounds X X)

N-{4-[(2-amino-6-methyl-4-pyrimidyl) amino] phenyl } acetamide hydrochloride (16): to 4-amino acetanilide 14 (3.55g, 23.64mmol) and 2-amino-4-chloro-6-methylpyrimidine 15 (3.73g, 26mmol) mixture in ethanol (50mL) adds dense HCl (2).Reaction mixture under refluxad stirred 2 hours, was cooled to 20 ℃ and filtration product, with more ethanol washing and dry, obtained 16 pure basically (6.38g, 92%); 203-207 ℃ of mp (EtOH); ¹H NMR[(CD ₃) ₂SO] δ 12.60 (br, 1H, N ⁺H), 10.50 (br 1H, NH), 10.02 (s, 1H, NH), 7.60-7.58 (br m, 6H, NH ₂﹠amp; ArH), 6.13 (s, 1H, H-5 "), 2.26 (s, 3H, CH ₃), 2.04 (s, 3H, CH ₃); HRMS (FAB ⁺), calculated value C ₁₃H ₁₆N ₅O (M ^{+ 1}) m/z 258.1355, measured value 258.1346.

N ⁴-(4-aminophenyl)-6-methyl-2,4-pyrimidinediamine hydrochloride (17): (6.0g, suspension 20mmol) add 2N HCl (40mL) and mixture were refluxed 20 hours to 16.Solvent evaporation to dry, seethed with excitement resistates and diluted with EtOAc in MeOH.The throw out that obtains is filtered and wash and drying, obtain 17 pure basically (5.0g, 97%) with EtOAc; 275-280 ℃ of mp (MeOH/EtOAc); ¹H NMR[(CD ₃) ₂SO] δ 12.50 (br, 1H, N ⁺H), 10.75 (br s, 1H, NH), 9.75 (br s, 2H, NH ₂), 7.85 (br s, 4H, NH ₂﹠amp; ArH)), 7.34 (d, J=8.6Hz, 2H, ArH), 6.24 (br s, 1H, ArH), 2.79 (s, 3H, CH ₃), HRMS (FAB ⁺) calculated value C ₁₁H ₁₄N ₅(M ^{+ 1}) m/z216.1249, measured value 216.1247.

N-{4-[(2-amino-6-methyl-4-pyrimidyl) amino] phenyl }-5-nitro-2-pyridine carboxamide 20.(1.06g, 6.31mmol) 1 hour (obtaining clear soln) of backflow in POC13 (10mL) is cooled to 20 ℃ also with excessive POCl to make 5-nitropyridine-2-carboxylic acid 18 ₃Under vacuum, remove.The resistates that obtains is dissolved in 1,4-diox ((20mL) and slowly join 17 (1.44g, 5.72mmol) and N, (2.0mL is 12.62mmol) 1, in the suspension in the 4-diox (20mL) for the N-Diethyl Aniline.Reaction mixture was stirred 3 days at 20 ℃.The white precipitate that obtains filtered and with more 1, the 4-diox washs.With solid at NH ₃Stir in the aqueous solution (20mL) and the red precipitate that obtains is filtered, wash with water and, obtain 20 (708mg, 31%) from the MeOH recrystallization; Mp (MeOH)〉290 ℃; ¹H NMR ([(CD ₃) ₂SO] and δ 10.74 (1H, NH), 9.43 (dd, J=2.6,0.5Hz, 1H, H-6), 8.96 (s, 1H, NH), 8.81 (dd, J=8.6,2.6Hz, 1H, H-4), 8.38 (dd, J=8.6,0.6Hz, 1H, H-3), 7.81 (d, J=9.0Hz, 2H, H-2 ' H-6 '), 7.70 (d, J=9.1Hz, 2H, H-3 ', 5 '), 6.09 (s, 2H, NH ₂), 5.87 (s, 1H, H-5 "), 2.09 (s, 3H, CH ₃); HRMS (FAB ⁺) calculated value C ₁₇H ₁₆N ₇O ₃(M ^{+ 1}) m/z366.1315, measured value 366.1314; Ultimate analysis calculated value C ₁₇H ₁₅N ₇O ₃.0.25 MeOH:C, 55.5; H, 4.4:N, 26.3; Measured value: C, 55.7; H, 4.5; N, 26.2%.

5-amino-N-{4-[(2-amino-6-methyl-4-pyrimidyl) amino] phenyl }-2-pyridine carboxamide (21): (523mg 1.43mmol) is suspended in that suspension among the 1:1 MeOH/THF (100ml) adds 10%Pd/C (100mg) and 55Hgmm hydrogenation 5 hours to 20.Reaction mixture is filtered and is evaporated to drying and from DCM/ sherwood oil (Pet.ether) recrystallization, obtains 21 (649mg, 96%); Mp (DCM/ sherwood oil)〉290 ℃; ¹H NMR[(CD ₃) ₂SO] δ 10.05 (s, 1H, NH), 8.85 (s, 1H, NH), 8.25 (d, J=2.5Hz, 1H, H-6), 7.82 (d, J=8.6Hz, 1H, H-3), 7.73 (d, J=9.0Hz, 2H, H-2 ', 6 '), 7.60 (d, J=9.0Hz, 2H, H-3 ', 5 '), 7.03 (dd, J=8.5,2.7Hz, 1H, H-4), 6.04 (br s, 2H, NH ₂), 6.03 (br s, 2H, NH ₂), 5.85 (s, 1H, H-5 "), 2.08 (s, 3H, CH ₃); Ultimate analysis calculated value C ₁₇H ₁₇N ₇).0.25 H ₂OC, 60.1; H, 5.2; N, 28.9; Measured value: C, 60.0; H, 5.2; N, 28.7%.

N-{4-[(2-amino-6-methyl-4-pyrimidyl) amino] phenyl }-5-(4-quinolyl amino)-2-pyridine carboxamide dihydrochloride (22) (compounds X X).(270mg is 0.81mmol) at EtOH (30mL) and H to 21 ₂Solution among the O (15mL) adds several dense HCl, and (264mg, 1.62mmol is 2eq) and 20 ℃ of stirrings, up to dissolving to add the 4-chloroquinoline subsequently.With reaction mixture refluxed 2 hours, add then more 4-chloroquinoline (264mg, 1.62mmol) and refluxed 20 hours.Reaction mixture dilutes with EtOAc, seethes with excitement and is cooled to 20 ℃.The throw out that obtains is filtered, obtain light yellow solid (it is 95% pure analyzing by HPLC).With this solid at NH ₃Stir in the aqueous solution.The throw out that obtains is filtered, wash with water, dry and from the MeOH recrystallization, obtain the free alkali (300mg) of product.(it is analyzed by HPLC is 98% purity).Following then free alkali is converted into HCl salt: add the MeOH (2.5mL) that contains 1.25M HCl, stirred 30 minutes, and be evaporated to drying.Resistates obtains 22 (SN 31319) (297mg 69%) from the MeOH/EtOAc recrystallization; HPLC99.2%; Mp (MeOH/EtOAc)〉290 ℃; ¹H NMR[(CD ₃) ₂SO] δ 15.00 (br, 1H, N ⁺H), 12.50 (br, 1H, N ⁺H), 11.23 (br s, 1H, NH), 10.74 (s, 1H, NH), 10.65 (br s, 1H, NH), 8.92 (d, J=2.3Hz, 1H, ArH), 8.89 (d, J=8.5Hz, 1H, ArH), 8.66 (d, J=6.8Hz, 1H, ArH), 8.31 (d, J=8.5Hz, 1H, ArH), 8.22 (dd, J=8.4,2.5Hz, 1H, ArH), 8.15 (d, J=7.9Hz, 1H, ArH), 8.08 (t, J=7.9Hz, 1H, ArH), 7.96 (d, J=8.9Hz, 2H, ArH), 7.87 (t, J=7.6Hz, 1H, ArH), 7.77 (br, 3H, ArH ﹠amp; NH ₂), 7.13 (d, J=6.8Hz, 1H, ArH), 6.18 (s, 1H, ArH), 2.28 (s, 3H, CH ₃) have an aromatic series CH signal not observe; Ultimate analysis calculated value C ₂₆H ₂₄Cl ₂N ₈O.0.5H ₂O:C, 57.4; H, 4.6; N, 20.6; Cl, 13.0; Measured value: C, 57.3; H, 4.8; N, 20.7; Cl, 12.7%.

Embodiment YY.

N-{6-[(2-amino-6-methyl-4-pyrimidyl) amino]-the 3-pyridyl }-preparation of 4-(4-quinolyl amino) benzamide dihydrochloride (compound YY)

6-methyl-N ⁴-(5-nitro-2-pyridyl)-2,4-pyrimidinediamine (9).To 2-amino-5-nitropyridine 1 (1.23g, 8.84mmol) and 2-amino-4-chloro-6-methylpyrimidine (1.40, g, ethanol 9.72mmol) (30mL) solution add several dense HCl.With reaction mixture refluxed 2 days, be cooled to 20 ℃ and with ethanol evaporation to dry.The brown jelly that obtains is stirred in MeOH, obtain filtrable throw out.With its filtration and with more MeOH washing, obtain product, be hydrochloride. ¹H NMR analyzes and shows that it is not fully pure.By with it at NH ₃Stirring in the aqueous solution, filtration and recrystallization are converted into free alkali, obtain 9 (682mg, 31%); Mp (MeO)〉300 ℃; ¹H NMR[(CD ₃) ₂SO] δ 10.47 (s, 1H, NH), and 9.10 (dd, J=2.8,0.4Hz, 1H, H-6 '), 8.39 (dd, J=9.4,2.8Hz, 1H, H-4 '), 8.28 (dd, J=9.3,0.3Hz, 1H, H-3 '), 6.63 (s, 1H, H-5), 6.39 (s, 2H, NH ₂), 2.18 (s, 3H, CH ₃); HRMS (EI ⁺) calculated value C ₁₀H ₁₀N ₆O ₂(M ⁺) m/z 246.0865, measured value 246.0866; Ultimate analysis calculated value C ₁₀H ₁₀N ₆O ₂: C, 48.8; H, 4.1; N, 34.1; Measured value C, 48.7; H, 4.2; N, 34.1%.

N ⁴-(5-amino-2-pyridyl)-6-methyl-2,4-pyrimidinediamine (10).(10% Pd/C (100mg) hydrogenation 20 hours under 45Hg mm is used in 634mg, 246mmol) hydrogenation in MeOH (50mL) with compound 9.Reaction mixture is filtered and be evaporated to drying, obtain 10 (550mg, 99%); 230-233 ℃ of mp (MeOH); ¹H NMR[(CD ₃) ₂SO] δ 8.96 (s, 1H, NH), and 7.70 (d, J=8.7Hz, 1H, H-3 '), 7.65 (d, J=2.5Hz, 1H, H-6 '), 6.95 (dd, J=8.8,2.9Hz, H-4 '), 6.30 (s, 1H, H-5), 5.94 (s, 2H, NH ₂), 4.84 (s, 2H, NH ₂), 2.07 (s, 3H, CH ₃); HRMS (FAB ⁺) calculated value C ₁₀H ₁₃N ₆(M ^{+ 1}) m/z 217.1202, measured value 217.1202; Ultimate analysis calculated value C ₁₀H ₁₂N ₆C, 55.5; H, 5.6; N, 38.9; Measured value C, 55.3; H, 5.7; N, 38.6%.

N-{6-[(2-amino-6-methyl-4-pyrimidyl) amino]-the 3-pyridyl }-4-nitrobenzamide (11).(500mg, 2.31mmol) and N, (1ml, 1.5eq) 1, the suspension in the 4-diox (20ml) drips paranitrobenzoyl chloride (429mg, solution 12.31mmol) to the N-Diethyl Aniline to 10 at 0 ℃.Reaction mixture was stirred 2 hours at 20 ℃.TLC and mass spectrum demonstration still have 10 to exist.Therefore add more paranitrobenzoyl chloride (43mg, 0.1eq) and stirred 20 hours.The throw out that obtains filtered and with more 1, the 4-diox washs.With the solid collected at NH ₃Stir in the aqueous solution, filter, wash with water and drying, obtain 11 pure basically (821mg, 97%); Mp (MeOH)〉300 ℃; ¹H NMR[(CD ₃) ₂SO] δ 10.58 (s, 1H, NH), 9.56 (s, 1H, NH), (8.65 d, J=2.5Hz, 1H, H-2 '), 8.38 (d, J=8.9Hz, 2H, H-3,5), 8.20 (d, J=9.2Hz, 2H, H-2,6), 8.17 (d, J=9.1Hz, 1H, H-5 '), (8.02 dd, J=9.0,2.6Hz, 1H, H-4 '), 6.45 (s, 1H, H-5 "), 6.15 (s, 2H, NH ₂), 2.12 (s, 3H, CH ₃); HRMS (FAB ⁺) (M ^{+ 1}) m/z calculated value C ₁₇H ₁₆N ₇O ₃(M ^{+ 1}) m/z, 366.1315, measured value 366.1314; Ultimate analysis calculated value C ₁₇H ₁₅N ₇O ₃: C, 55.9; H, 4.1, N, 26.8, measured value C, 55.6; H, 4.2; N, 26.8%.

4-amino-N-{6-[(2-amino-6-methyl-4-pyrimidyl) amino]-the 3-pyridyl } benzamide 12.(790mg, 2.16mmol) suspension in 1:1 MeOH/THF (100ml) adds 10% Pd/C (100mg) and hydrogenation 20 hours under 45Hgmm to 11.Reaction mixture is filtered and is evaporated to drying and from DCM/ sherwood oil recrystallization, obtains 12 (695mg, 96%); Mp (DCM/ sherwood oil)〉300 ℃; ¹H NMR[(CD ₃) ₂SO] δ 9.78 (s, 1H, NH), 9.45 (s, 1H, NH), (8.63 dd, J=2.4,0.3Hz, 1H, H-2 '), 8.09 (d, J=9.0Hz, 1H, H-5 '), 7.98 (dd, J=9.0,2.6Hz, 1H, H-4 '), 7.72 (d, J=8.7Hz, 2H, H-2,6), 6.61 (d, J=8.7Hz, 2H, H-3,5), 6.43 (s, 1H, H-5 "), 6.13 (s, 2H, NH ₂), 5.73 (s, 2H, NH ₂), 2.12 (s, 3H, CH ₃); HRMS (FAB ⁺) calculated value C ₁₇H ₁₈N ₇O (M ^{+ 1}) m/z336.1573, measured value 336.1578; Ultimate analysis calculated value C ₁₇H ₁₇N ₇O.0.25 H ₂O:C, 60.1; H, 5.2; N, 28.9; C, 60.2; N, 5.3; N, 29.0%.

N-{6-[(2-amino-6-methyl-4-pyrimidyl) amino]-the 3-pyridyl }-4-(4-quinolyl amino) benzamide dihydrochloride (13) (compound YY).(250mg is 0.75mmol) at EtOH (40mL) and H to 12 ₂Solution among the O (20mL) adds several dense HCl, and (159mg, 0.98mmol is 1.3eq) and 20 ℃ of stirrings, up to dissolving to add the 4-chloroquinoline subsequently.With reaction mixture refluxed 4 hours, add more 4-chloroquinoline (100mg) then and refluxed 20 hours.Reaction mixture dilutes with EtOAc, seethes with excitement and is cooled to 20 ℃.The throw out that obtains is filtered, obtain light yellow solid (analyzing by HPLC is 75% purity).With this solid at NH ₃Stir in the aqueous solution.The throw out that obtains is filtered, wash with water and drying, obtain the free alkali of product.It is analyzed by HPLC is 96% purity, following then free alkali is converted into HCl salt: add the MeOH (2.5mL) that contains 1.25M HCl, stirred 30 minutes, and be evaporated to drying.Resistates stirs in MeOH (10mL), filters and drying, obtains 13 (compound YY) (365mg91%); HPLC 98.9%; Mp (MeOH)〉290 ℃; ¹H NMR[(CD ₃) ₂SO] δ 14.70 (br1H, N ⁺H), 12.98 (br, 1H, N ⁺H), 11.12 (s, 1H, NH), 11.01 (s, 1H, NH), 10.68 (s, 1H, NH), 8.88 (d, J=3.0Hz, 1H, ArH), 8.84 (d, J=8.6Hz, 1H, ArH), 8.62 (d, J=6.9Hz, 1H, ArH), 8.26 (dd, J=9.0,2.6Hz, 1H, ArH), 8.22 (d, J=8.6Hz, 2H, ArH), 8.15-8.05 (m, 2H, ArH), 7.86 (ddd, J=7.6,6.8,1.5Hz, 1H, ArH), 7.70 (d, J=8.6Hz, 2H, ArH), 7.01 (d, J=6.9Hz, 1H, ArH), 7.02 (v.br, 1H, ArH), 2.32 (s, 3H, CH ₃), NH ₂Signal and an aromatic signal do not observe; HRMS (FAB ⁺) calculated value C ₂₆H ₂₃N ₈O (M ^{+ 1}) m/z 463.1995, measured value 463.1996; Ultimate analysis calculated value C ₂₆H ₂₄N ₈Cl ₂O.HCl.0.25H ₂O:, C, 54.2; H, 4.5; N, 19.4; Cl, 18.5; Measured value C, 54.2; H, 4.5; N, 19.3; Cl, 17.7%.

Embodiment ZZ.

N-{5-[(2-amino-6-methyl-4-pyrimidyl) amino]-the 2-pyridyl }-preparation of 4-(4-quinolyl amino) benzamide dihydrochloride (compound ZZ)

N-{5-[(2-amino-6-methyl-4-pyrimidyl) amino]-the 2-pyridyl } ethanamide 4.(1.04g, 6.88mmol) (1.09g, EtOH 7.57mmol) (30mL) solution adds 2 dense HCl with 2-amino-4-chloro-6-methylpyrimidine to N-(5-amino-2-pyridyl) ethanamide 2.With reaction mixture refluxed 2 hours, be cooled to 20 ℃.The throw out that obtains is filtered,, obtain (1.85g), obtain other material (165mg) by the mother liquor concentrating and separating with more ethanol washing and dry.Total recovery 499.3%.mp (MeOH/EtOAc)〉300 ℃. ¹HNMR[(CD ₃) ₂SO] δ 12.77 (br, 1H, NH), 10.72 (br, 1H, NH), 10.44 (s, 1H, NH), 8.63 (br s, 1H, ArH), 8.13-8.06 (m, 2H, ArH), 7.79 (v.br, 2H, NH ₂), 6.18 (s, 1H, ArH), 2.30 (s, 3H, COCH ₃), 2.09 (s, 3H, CH ₃); HRMS (FAB ⁺) calculated value C ₁₂H ₁₅N ₆O (M ^{+ 1}) m/z 259.1307, measured value 259.1304; Ultimate analysis calculated value C ₁₂H _15C1N ₆O.H ₂O:C, 46.1; H, 5.5; N, 26.9; Cl, 11.3; Measured value C, 45.9; H, 5.5; N, 26.4; Cl, 11.45%.

N ⁴-(6-amino-3-pyridyl)-6-methyl-2,4-pyrimidinediamine 5.With 4 (1.53g, 5.19mmol) 1,4-diox/MeOH (1:1,100mL) and 2N HCl[10mL, H ₂The dense HCl 2mL of OmL+)] suspension returning in 24 hours.With solvent evaporation to dry and with resistates NH ₃Aqueous solution alkalization.(10 * 50mL) extract the solution that obtains, dry (Na with EtOAc ₂SO ₄) and evaporating solvent, obtain 5 (1.11g, 99%).Little sample is from DCM/ sherwood oil recrystallization; 183-186 ℃ of mp (DCM/ sherwood oil); ¹H NMR[(CD ₃) ₂SO] δ 8.41 (s, 1H, NH), 8.01 (d, J=2.5Hz, 1H, H-2 '), 7.56 (dd, J=8.7, '), 6.42 (d, ' 2.6Hz, 1H, H-4J=8.8Hz, 1H, H-5 '), 5.90 (s, 2H, NH ₂), 5.67 (s, 1H, H-5), 5.59 (s, 2H, NH ₂H, CH), 2.03 (s, 3 ₃) HRMS (EI ⁺) calculated value C ₁₀H ₁₂N ₆(M ⁺) m/z 216.1123, measured value 216.1124; Ultimate analysis calculated value C ₁₀H ₁₂N ₆.0.25H ₂O:C, 54.4; H, 5.7; N, 38.1; Measured value: C, 54.4; H, 5.7; N, 37.9%.

N-{5-[(2-amino-6-methyl-4-pyrimidyl) amino]-the 2-pyridyl }-the 4-nitrobenzamide) 6.0 ℃ to 5 (830mg, 3.84mmol) and N, the N-Diethyl Aniline (1.0mL, 5.76mmol) 1, the suspension in the 4-diox (20mL) drip the 4-nitrobenzoyl chloride (720mg, 3.88mmol) 1,4-diox (20mL) solution.After adding is finished, reaction mixture was stirred 1 hour at 20 ℃, then diox is removed under vacuum.Resistates is at H ₂Stir among the O (50mL) and the throw out that obtains is filtered, use NH ₃The aqueous solution, H ₂O and petroleum ether.Resistates seethes with excitement in MeOH and collects undissolved red solid.This handles triplicate again, obtains 6 pure basically (903mg, 65%), [if repeat, product Cong diox should be filtered out, to avoid extra step]; 294-297 ℃ of mp (MeOH); ¹H NMR[(CD ₃) ₂SO] 10.99 (s, 1H, NH), 9.14 (s, 1H, NH), 8.68 (d, J=2.5Hz, 1H, H-6 '), 8.32 (d, J=8.9Hz, 2H, H-3﹠amp; 5), 8.27 (dd, J=9.0,2.7Hz, 1H, H-4 '), 8.23 (d, J=8.9Hz, 2H, H-2﹠amp; H-6), 8.08 (d, J=9.0Hz, 1H, H-3 '), 6.18 (s, 2H, NH ₂), 5.88 (s, 1H, H-5 "), 2.11 (s, 3H, CH ₃); HRMS (FAB ⁺), calculated value C ₁₇H ₁₆N ₇O ₃(M ^{+ 1}) m/z366.1315, measured value 366.1312; Ultimate analysis calculated value C ₁₇H ₁₅N ₇O ₃.CH ₃OH:C, 54.4; H, 4.8; N, 24.7; Measured value C, 54.5; H, 4.8; N, 24.7%.

4-amino-N-{5-[(2-amino-6-methyl-4-pyrimidyl) amino]-the 2-pyridyl } benzamide 7.(802mg, 2.19mmol) 1: the suspension among the 1MeOH/THF (100ml) adds 10%Pd/C (100mg) and hydrogenation 20 hours under 45Hgmm to 6.Reaction mixture is filtered and is evaporated to drying and from DCM/ sherwood oil recrystallization, obtains 7 (697mg, 95%); 157-161 ℃ of mp (DCM/ sherwood oil); ¹H NMR[(CD ₃) ₂SO] δ 9.98 (s, 1H, NH), 9.03 (s, 1H, NH), (8.59 d, J=2.4Hz, 1H, H-6 '), 8.16 (dd, J=9.0,2.7Hz, 1H, H-4 '), 8.04 (d, J=8.9Hz, 1H, H-3 '), 7.77 (d, J=8.7Hz, 2H, H-2,6), 6.57 (d, J=8.7Hz, 2H, H-3,5), 6.13 (s, 2H, NH ₂), 5.13 (s, 1H, H-5 "), 5.74 (s, 2H, NH ₂), 2.10 (s, 3H, CH ₃); HRMS (FAB ⁺) calculated value C ₁₇H ₁₈N ₇O (M ^{+ 1}) m/z336.1573, measured value 366.1574; Ultimate analysis calculated value C ₁₇H ₁₇N ₇O.2H ₂O:C, 55.0; H, 5,7; N, 26.4; Measured value C, 55.2; H, 5.7; N, 26.3%.

N-{5-[(2-amino-6-methyl-4-pyrimidyl) amino]-the 2-pyridyl }-4-(4-quinolyl amino) benzamide dihydrochloride 8 (compound ZZ).(241mg is 0.72mmol) at EtOH (20mL) and H to 7 ₂Solution among the O (10mL) adds the 4-chloroquinoline, and (153mg, 0.94mmol is 1.3eq) and 20 ℃ of stirrings, up to dissolving.Drip several dense HCl then and refluxed 24 hours.Reaction mixture dilutes with EtOAc, seethes with excitement and is cooled to 20 ℃.The throw out that obtains is filtered, obtain light yellow solid, it from the MeOH/EtOAc recrystallization, is obtained the product of 350mg, it is analyzed by HPLC is 84% purity.With this solid at NH ₃Stir in the aqueous solution so that be converted into the free alkali of product, throw out is filtered, wash with water and drying, obtain 204mg.It is analyzed by HPLC is 98% purity.Following free alkali is converted into HCl salt: add the MeOH (1mL) that contains 1.25M HCl, stirred 30 minutes, filter and drying, obtain 8 (compound ZZ) (224mg 58%); HPLC 100%; Mp (MeOH)〉295 ℃; ¹H NMR[(CD ₃) ₂SO] δ 14.75 (br, 1H, N ⁺H), 12.89 (bs, 1H, N ⁺H), 11.14 (s, 1H, NH), 10.99 (bs, 1H, NH), 10.94 (s, 1H, NH), 8.87 (d, J=8.5Hz, 1H, ArH), 8.77 (br1H, ArH), 8.61 (d, J=6.9Hz, 1H, ArH), 8.31 (br, 1H, ArH), 8.23 (d, J=8.6Hz, 2H, ArH), 8.13 (dd, J=8.0,0.9Hz, 1H, ArH), 8.07 (td, J=7.7,0.9Hz, 1H, ArH), 7.85 (td, J=7.7,1.2Hz, 1H, ArH), 7.67 (d, J=8.6Hz, 2H, ArH), 7.02 (d, J=6.9Hz, 1H, ArH), 6.26 (s, 1H, ArH), 2.31 (s, 3H, CH ₃); HRMS (FAB+), calculated value C ₂₆H ₂₃N ₈O (M ^{+ 1}) m/z 4631995, measured value 463.1994; Ultimate analysis calculated value C26H24Cl2N80.HCl.H ₂O:C, %2.9; H, 4.6; N, 19.0; Cl, 18.0; Measured value C.53.1; H, 4.6; N, 19.2; Cl, 17.9%.

Embodiment A AA

4-(pyridin-4-yl sulfenyl) aniline (3).To 4-amino-benzene mercaptan [1] (5.10g, dry DMF 40.74mmol) (90mL) solution sequentially add 4-chloropyridine hydrochloride [2] (6.41g, 42.68mmol) and anhydrous K ₂CO ₃(14.70g 106.34mmol), and makes the suspension that obtains in room temperature vigorous stirring～3 hour.After this, reaction mixture EtOAc (100mL) and H ₂O (100mL) dilutes, and organic layer is separated.Water layer further extracts (100mL x 2) with EtOAc, then organic extraction is merged, and uses the salt water washing, and uses anhydrous MgSO at last ₄Dry.Removal of solvent under reduced pressure, and use 1:1 Et ₂O: the resistates that hexane (300mL) washing obtains, obtain amine 3, be thin canescence crystalline solid (4.64g, 56%) mp (MeOH: EtOAc) 171-173 ℃; ¹H NMR[(CD ₃) ₂SO]: δ 5.64 (s, 2H, NH ₂), 6.67 (ddd, J=9.40,4.78,2.81Hz, 2H, ArH), 6.90 (dd, J=4.61,1.59Hz, 2H, ArH), 7.20 (ddd, J=9.40,4.78,2.81Hz, 2H, ArH), 8.29 (dd, J=4.61,1.59Hz, 2H, ArH); HRMS: calculated value C ₁₁H ₁₁N ₂S (M ⁺) 203.0643, measured value 203.0641.

4-nitro-N-[4-(pyridin-4-yl sulfenyl) phenyl] benzamide (5).To amine 3 (2.03g, 10.06mmol) no Shui diox (70mL) solution sequentially add anhydrous pyridine (4.05mL, 50.28mmol) and 4-nitrobenzoyl chloride (4) (3.19g, 17.18mmol, be the solution in the no Shui diox of 30mL), and the mixture that obtains stirred 14 hours at～50 ℃.After this, the yellow solid that obtains by filtering separation, and with its sequentially Yong diox, EtOAc and hexane wash.With the solid that obtains be dissolved in again MeOH (～5L), and this solution filtered by diatomite to remove undissolved impurity, concentrating under reduced pressure is littler volume then.By filter collecting the solid that obtains, and then suspend and, and regather by filtration at last sequentially with EtOH, MeOH and EtOAc washing.The material that obtains obtains acid amides 5 with hexane wash and high vacuum dry, is unbodied yellow powder shape solid, mp 295-298 ℃; ¹H NMR[(CD ₃) ₂SO]: δ 7.38 (d, J=6.15Hz, 2H, ArH), 7.55 (ddd, J=9.42,4.45,2.59Hz, 2H, ArH), 8.07 (m, 2H, ArH), 8.24 (ddd, J=9.21,4.32,2.31Hz, 2H, ArH), 8.39 (dd, J=6.92,1.97Hz, 2H, ArH), 8.53 (d, J=6.6Hz, 2H, ArH), 10.97 (s, 1H ,-C (O) NH-); HRMS: calculated value C ₁₈H ₁₄N ₃O ₃S (M ⁺) 352.0756, measured value 352.0755.

4-amino-N-[(4-(pyridin-4-yl sulfenyl) phenyl] benzamide (6).To the nitro-compound 5 (0.54g, 2:1 EtOH:H 1.53mmol) that reflux ₂(0.54g 9.72mmol) and dense HCl (2mL), and made the suspension returning that obtains 1 hour sequentially to add the Fe powder in O (100mL) solution.After this, Celite pad is passed through in the reaction mixture filtration of heat, and the solvent decompression is removed.Resistates is dissolved in MeOH again, and the solution that obtains is stirred with diatomite spend the night.The suspension filtered that obtains is passed through Celite pad, and with the hcl acidifying in the filtrate usefulness 1.25M methyl alcohol.The volume that the solution decompression simmer down to is littler, and remove the solid that obtains by Celite pad by filtering.With filtrate acidifying once more, and processing (twice) as previously mentioned,, obtaining unbodied tawny solid (0.38g, 77%) at last, it is without being further purified use; ¹H NMR[(CD ₃) ₂SO]: δ 5.79 (s, 2H ,-NH ₂), 6.62 (d, J=8.58Hz, 2H, ArH), 6.98 (d, J=5.96Hz, 2H, ArH), 7.53 (d, J=8.61Hz, 2H, ArH), 7.74 (d, J=8.58Hz, 2H, ArH), 7.96 (d, J=8.61Hz, 2H, ArH), 8.34 (d, J=5.35Hz, 2H, ArH), 10.02 (s, 1H ,-C (O) NH-); LCMS (APCI ⁺): 322 (100%).

N-[4-(pyridin-4-yl sulfenyl) phenyl]-4-(quinolyl-4 amino) benzamide hydrochloride salt (compd A AA).To amine 5 (0.31g, 0.97mmol) solution in the 20% EtOH aqueous solution (100mL) sequentially add 4-chloroquinoline [7] (0.33g, 2.02mmol) and dense HCl (0.20mL 8.71mmol), and made the suspension returning that obtains 15 hours.After this, the solvent decompression is removed, and by two MeOH-azeotropic cyclic drying resistatess.The solid that obtains is by the column chromatography on the silica gel (twice) purifying, with 5% → 10% → 20%MeOH:CH ₂Cl ₂Wash-out obtains solid residue, with it from MeOH: the hydrochloric acid the methyl alcohol: the EtOAc redeposition, obtain compd A AA, be unbodied yellow solid (0.15g, 29%), 306-310 ℃ of mp (EtOAc:MeOH); ¹H NMR[(CD ₃) ₂SO]: δ 7.01 (d, J=6.94Hz, 1H, ArH), 7.45 (d, J=6.77Hz, 2H, ArH), 7.70 (m, 4H), 7.86 (m, 1H), 8.11 (m, 4H), 8.22 (dd, J=6.82,1.79Hz, 2H, ArH), 8.55 (d, J=6.77Hz, 2H, ArH), 8.62 (d, J=6.94Hz, 1H, ArH), 8.89 (d, J=8.35Hz, 1H, ArH), 10.78 (s, 1H, ArNHAr), (11.19 s, 1H ,-C (O) NH-), 14.72 (brs, 1H, pyridine-N ⁺-H) [quinoline N ⁺-H is invisible]; HRMS: calculated value C ₂₇H ₂₁N ₄OS (M ⁺) 449.1436, measured value 449.1441; HPLC:99.3%.

Embodiment B BB

4-(4-aminophenyl sulfenyl) pyridine-2-amine (9).To 4-amino-benzene mercaptan (1) (9.49g, dry DMF 75.77mmol) (54mL) solution sequentially add 4-chloro-2-aminopyridine (8) (3.70g, 28.80mmol) and anhydrous K ₂CO ₃(10.70g 107.97mmol), and stirs the yellow suspension that obtains～45 minutes～120 ℃ (bathing temperature).After this, with the brown-black suspension cool to room temperature that obtains, use H then ₂O and EtOAc dilution.The mixture that obtains extracts with EtOAc (x2), organic fraction is merged and uses the salt water washing, uses MgSO then ₄Dry.The solvent decompression is removed, then resistates is dissolved among the MeOH of a little volume again, and filter and pass through silicagel pad.The solvent decompression is removed, obtain amine 9 (5.66g, 90%), be unbodied butteriness purple solid, mp:141-143 ℃; ¹H NMR[(CD ₃) ₂SO]: δ 5.56 (br s, 2H, NH ₂), 5.76 (br s, 2H, NH ₂), 5.94 (d, J=1.22Hz, 1H, ArH), 6.10 (dd, J=5.46,1.68Hz, 1H, ArH), 6.64 (ddd, J=9.37,4.78,2.79Hz, 2H, ArH), 7.17 (ddd, J=9.38,4.74,2.79Hz, 2H, ArH), 7.65 (d, J=5.44Hz, 1H, ArH); HRMS: calculated value C ₁₁H ₁₂N ₃S (MH ⁺) m/z218.0753, measured value 218.0750.

N-[4-(2-aminopyridine-4-base sulfenyl) phenyl]-4-nitrobenzamide (10).(1.09g, no Shui diox (30mL) solution 5.01mmol) sequentially add anhydrous pyridine, and (2.08mL is 25.04mmol) with 4-nitrobenzoyl chloride (4) (1.60g, 8.60mmol to amine 9; Add as the solution in the 20mL dry DMF), and the mixture that obtains stirred～4 hours 55-60 ℃ (bathing temperature).After this, collect the solid that obtains by filtering, and incite somebody to action sequentially Yong diox, EtOAc and hexane wash.Crude product is from MeOH: the hydrochloric acid the methyl alcohol: the EtOAc redeposition obtain nitro-compound 10, is unbodied yellow solid (1.19g, 64%), mp〉300 ℃; ¹H NMR[(CD ₃) ₂SO]: δ 13.35 (brs, 1H, quinoline-N ⁺-H), 10.97 (s, 1H, ArC (O) NHAr), 8.39 (ddd, J=9.25,4.40,2.37Hz, 2H, ArH), 8.23 (ddd, J=9.20,4.34,2.31HZ, 2H, ArH), 8.06 (ddd, J=9.43,4.53,2.62Hz, 2H, ArH), 7.81 (m, 3H, ArH ﹠amp; ArNH ₂), 7.66 (ddd, J=9.41,4.52,2.61Hz, 2H, ArH), 6.65 (dd, J=6.87,1.91Hz, 1H, ArH), 6.29 (d, J=1.73Hz, 1H, ArH); HRMS: calculated value C ₁₈H ₁₅N ₄O ₃S (MH ⁺) m/z367.0865, measured value 367.0865.

4-amino-N-[4-(2-aminopyridine-4-base sulfenyl) phenyl] benzamide hydrochloride salt (11).(0.83g 2.07mmol) is suspended in 2:1 EtOH:H with nitro-compound 10 ₂Among the O (100mL) and with the suspension returning that obtains.(0.54g, 9.59mmol) with dense HCl (2mL), and the dark orange suspension that will obtain refluxed 1 hour sequentially to add the Fe powder to this mixture.After this, the yellow suspension filtered while hot that obtains is passed through Celite pad, and the solvent decompression is removed.Resistates is resuspended in H ₂On the O, to wherein adding a large amount of diatomite, and the suspension that obtains stirred spend the night.After this, this suspension filtered is passed through Celite pad, and the solvent decompression is removed, obtain thick amine 11, be unbodied pale solid (0.65g, 93%) that it is without using through single step purification. ¹H NMR[(CD ₃) ₂SO]: δ 9.98 (s, 1H, ArC (O) NHAr), 7.91 (ddd, J=9.40,4.48,2.58Hz, 2H, ArH), 7.72 (m, 3H, ArH), 7.48 (ddd, J=9.37,4.39,2.55Hz, 2H, ArH), 6.61 (d, J=7.85Hz, 2H, ArH), 6.18 (dd, J=5.46,1.68Hz, 1H, ArH), 5.82 (d, J=1.36Hz, 1H, ArH), 6.01 (br s, 2H, ArNH ₂), 5.82 (br s, 2H, ArNH ₂); HRMS: calculated value C ₁₈H ₁₇N ₄OS (MH ⁺) m/z 337.1123, measured value 337.1126.

N-[4-(2-aminopyridine-4-base sulfenyl) phenyl]-4-(quinolyl-4 amino) benzamide hydrochloride salt (compd B BB).(0.66g, 4.01mmol) (0.52mL, (0.63g 1.88mmol) refluxed 3 hours in the solution in the 20% EtOH aqueous solution (100mL) and with the mixture that obtains 17.13mmol) sequentially to join amine 11 with dense HCl with 4-chloroquinoline (7).After this, the solvent decompression is removed, and resistates is passed through two MeOH-azeotropic cyclic dryings.Make resistates from MeOH then: the hydrochloric acid the methyl alcohol: EtOAc redeposition twice, obtain compd B BB (0.55g, 54%), be unbodied yellow solid, mp 225-239 ℃; ¹H NMR[(CD ₃) ₂SO]: δ 14.10 (vvbrs, 2H, quinolyl-N ⁺-H﹠amp; Pyridyl-N ⁺H), 11.14 (s, 1H, ArNHAr), 10.75 (s, 1H, ArC (O) NHAr), 8.87 (d, J=8.48Hz, 1H, ArH), 8.62 (d, J=6.92Hz, 1H, ArH), 8.20 (d, J=8.61Hz, 2H, ArH), 8.09 (m, 4H, ArH), 7.82 (m, 4H, ArH ﹠amp; ArNH ₂), 7.68 (m, 4H, ArH), 7.01 (d, J=6.91Hz, 1H, ArH), 6.66 (dd, J=6.88,1.87Hz, 1H, ArH), 6.31 (d, J=1.74Hz, 1H, ArH); HRMS: calculated value C ₂₇H ₂₂N ₅OS (MH ⁺) m/z 464.1541, measured value 464.1541; HPLC:97.4%.

Embodiment C CC

4-(4-aminophenyl sulfenyl) pyridine-2-alcohol (13).To 4-amino-benzene mercaptan [1] (3.68g, dry DMF 29.40mmol) (54mL) solution sequentially add 4-chloro-2 hydroxy pyrimidine (12) (0.49g, 3.81mmol) and anhydrous K ₂CO ₃(10.70g 107.97mmol), and stirs the yellow suspension that obtains～1 hour～120 ℃ (bathing temperature).Afterwards, the demonstration of the lcms analysis of reaction mixture still has many 12 to exist, and therefore adds more 1 (1.28g, 10.22mmol) (as the adding of the solution in the 10mL dry DMF) of volume.After 1 hour, LCMS and TLC analyze the demonstration reaction and finish, and therefore with the reaction mixture cool to room temperature, use H ₂O dilutes, and extracts (x3) with EtOAc.Anhydrous MgSO is used in the organic extraction salt water washing that merges ₄Drying, and solvent decompression removed.Resistates is by the column chromatography purifying on the silica gel, with 1% → 10%MeOH:CH ₂Cl ₂Wash-out obtains amine 13, is unbodied pale solid (0.47g, 56%) that it is without being further purified use; ¹H NMR[(CD ₃) ₂SO]: δ 11.20 (br s, 1H, ArOH), 7.18 (m, 3H, ArH), 6.66 (ddd, J=9.38,4.76,2.80Hz, 2H, ArH), 5.89 (dd, J=6.95,1.92Hz), 5.64 (brs, 2H, ArNH ₂), 5.55 (d, J=1.79Hz, 1H, ArH); HRMS: calculated value C ₁₁H ₁₁N ₂OS (MH ⁺) m/z 219.0592, measured value 219.0591.

N-[4-(2 hydroxy pyrimidine-4-base sulfenyl) phenyl]-4-nitrobenzamide (14).To amine 13 (0.47g, 2.13mmol) no Shui diox (120mL) solution sequentially add anhydrous pyridine (0.86mL, 10.65mmol) and 4-nitrobenzoyl chloride (4) (0.70g, 3.76mmol, add as the solution in the 20mL dry DMF), and the mixture that obtains spent the night stirring～50 ℃ (bathing temperature).After this, with the reaction mixture cool to room temperature, merge with a large amount of silicon-dioxide then.Solvent decompression is removed, and with the silicon-dioxide adsorptive that obtains by the column chromatography purifying on the silica gel, with 1% → 20% MeOH:CH ₂Cl ₂(contain 0.5%NH ₃The aqueous solution) wash-out obtains purer 14.This material washs with MeOH, and undissolved solid filtering is fallen and drying, obtains first nitro-compound 14 (0.27g).Filtrate decompression is concentrated, and with the MeOH washing of resistates with a little volume, dry then 14 (0.30g adds up to 68%) that obtain other amount, mp 255-260 ℃ (dark powder → tarry materials); ¹H NMR[(CD ₃) ₂SO]: δ 10.86 (s, 1H, ArC (O) NHAr), 8.38 (ddd, J=9.22,4.34,2.33Hz, 2H ArH), 8.22 (ddd, J=9.24,4.32,2.31Hz, 2H, ArH), (7.98 ddd, J=9.40,4.48,2.57Hz, 2H ArH), 7.61 (ddd, J=9.33,4.46,2.56Hz, 2H, ArH), 7.38 (d, J=6.95Hz, 1H, ArH), 6.05 (dd, J=6.94,1.93Hz, 1H, ArH), 5.73 (d, J=1.78Hz, 1H, ArH) [the OH signal is at about 5.6ppm place, non-constant widths]; HRMS: calculated value C ₁₈H ₁₄N ₃O ₄S (MH ⁺) m/z 368.0705, measured value 368.0711.

4-amino-N-[4-(2 hydroxy pyrimidine-4-base sulfenyl] phenyl) benzamide (15).(0.48g 1.18mmol) is suspended in 2:1 EtOH:H with nitro-compound 14 ₂Among the O (100mL) and with the suspension returning that obtains.(0.30g 9.59mmol) and dense HCl (2mL), and made the suspension returning that obtains 15 minutes sequentially to add the Fe powder to this mixture.At this moment, TLC and lcms analysis show reaction not exclusively, and (0.80g 14.29mmol) and dense HCl (10mL), and refluxes other mixture 50 minutes therefore to add the Fe powder of other amount.After this, reaction is finished, and thus, the reaction mixture heat filtering is passed through Celite pad, and the solvent decompression is removed.Resistates is by two MeOH-azeotropic cyclic dryings, and then is dissolved among the MeOH and stirs with diatomite and gac and spend the night.The soup compound that obtains is filtered by Celite pad, and the solvent decompression is removed.Resistates be dissolved in MeOH again and be adsorbed onto on a large amount of silica gel, and with the silicon-dioxide adsorptive that obtains by the column chromatography purifying on the silica gel, with 1% → 20% MeOH:CH ₂Cl ₂(contain 0.5%NH ₃The aqueous solution) wash-out obtains thick 15.This material is further by from MeOH: the hydrochloric acid the methyl alcohol: the EtOAc redeposition comes purifying, obtain amine 15, for unbodied cream-colored solid (15mg, 3%-may be owing to be adsorbed in gac and cause many material unaccounted-for (MUF)s, gac is extracted several times, what does not obtain); ¹H NMR[(CD ₃) ₂SO]: δ 11.40 (v v br s, 1H, quinolyl-N ⁺-H), 10.08 (s, 1H, ArH), 7.94 (ddd, J=9.40,4.51,2.59Hz, 2H, ArH), 7.78 (d, J=8.67Hz, 2H, ArH), 7.52 (ddd, J=9.40,4.49,2.61Hz, 2H, ArH), 7.27 (d, J=6.95Hz, 1H, ArH), 6.71 (d, J=8.52Hz, 2H, ArH), 5.96 (dd, J=6.96,1.93Hz, 1H, ArH) [ArOH ﹠amp; ArNH ₂Invisible]; LCMS (APCI ⁺): 338 (100%), 423 (60%), 169 (40%).

N-[4-(2 hydroxy pyrimidine-4-base sulfenyl) phenyl]-4-(quinolyl-4 amino) benzamide hydrochloride salt (Compound C CC).With 4-chloroquinoline (7) (12mg, 0.07mmol) and dense HCl (30 μ L, (13mg 0.032mmol) in the solution in the 20% EtOH aqueous solution (6mL), and refluxes the mixture that obtains 18 hours 17.13mmol) sequentially to join amine 15.After this, the solvent decompression is removed, and resistates is passed through two MeOH-azeotropic cyclic dryings.The material that obtains be dissolved in MeOH again and be adsorbed onto on a large amount of silica gel, and with the silicon-dioxide adsorptive that obtains by the column chromatography purifying on the silica gel, with 1% → 10%MeOH:CH ₂Cl ₂(contain 0.5%NH ₃The aqueous solution) wash-out obtains purer material.With this material further by from MeOH: the hydrochloric acid the methyl alcohol: the EtOAc redeposition comes purifying, obtains Compound C CC, is unbodied yellow solid (7mg, 41%), mp 209-214 ℃; ¹H NMR[(CD ₃) ₂SO]: δ 14.42 (br s, 1H, 1H, quinolyl-N ⁺-H), 11.34 (br s, 1H, pyridyl-N ⁺-H), 10.99 (s, 1H, ArNHAr), 10.61 (s, 1H, ArC (O) NHAr), 8.77 (d, J=8.60Hz, 1H, ArH), 8.62 (d, J=6.90Hz, 1H, ArH), 8.19 (d, J=8.60Hz, 2H, ArH), 8.07 (m, 2H), 8.00 (dd, J=6.88,1.83Hz, 2H, ArH), 7.87 (septet, J=11.29,8.36,5.40,2.76Hz, 1H, ArH), 7.69 (d, J=8.55Hz, 2H, ArH), 7.60 (d, J=8.60Hz, 2H, ArH), 7.27 (d, J=6.97Hz, 1H, ArH), 7.06 (d, J=6.94Hz, 1H, ArH), 5.97 (d, J=6.96,1.88Hz), 1H, ArH), 5.63 (d, J=1.76Hz, 1H, ArH) [ArOH is invisible]; HRMS: calculated value C ₂₇H ₂₁N ₄O ₂S (MH ⁺) m/z 465.1385, measured value 465.1389; HPLC:93.5%.

Embodiment DDD

The biological activity of The compounds of this invention

Of people such as Ghoshal [Mol.Cell Biol.2005,11,4727-41], the antibody of the high degree of specificity of all three kinds of enzymes of use antagonism carries out being made by 4-anilino quinoline the test of DNMT1 selectivity consumption.For DNMT1, use the commercially available antibody that derives from SaNta Cruz or NewENglaNd Biolabs.Antibody with high-titer antagonism DNMT3a and 3b is freshly prepd, and described antibody can intersected with each otherly not react in western blotting or immunoprecipitation analysis.From being HCT116 cell (with colon cancer cell) the isolated protein extract (being equivalent to 100 μ g) that the 4-anilino quinoline of 5,10 and 100 μ M is handled than all three kinds of DNMT of higher horizontal expression with concentration.As shown in table 3, in the compound of screening, have nine compounds at 100 μ M, 13 compounds are arranged at 10 μ M with there are 11 compounds can induce greater than 60% DNMT1 degraded (the DNMT1 level is lower than 40%) at 5 μ M.Four compounds (Compound D DD1, Compound D DD2, Compound D DD3 and Compound D DD4) can induce the DNMT1 greater than 94% to degrade (the DNMT1 level is lower than 6%) at 10 μ M, show other effort levels of level that is realized by Decitabine.

The demethylation activity of the 4-anilino quinoline of the non--two quaternary ammoniums of test in based on GFP (green fluorescent protein) test of cell.This test has the GFP gene regulated by the CMV promotor and to the sensitivity that methylates of the CpG position in the promotor.Because be exposed to that methylation inhibitor causes methylate to reduce causes GFP to express, and easily estimates.Particularly, the reactivate that uses the genetically modified CMV-EE210 clone test of the GFP that comprises outer genetic silencing GFP to express by the stream cell counting.By with pTR-UF/UF1/UF2 plasmid (people such as ZolotuhiN, 1996) transfection NIH 3T3 cell preparation CMV-EE210, described pTR-UF/UF1/UF2 plasmid is by pBS (+) (StratageNe that comprises cytomegalovirus (CMV) promotor, INc.) form, described cytomegalovirus promoter causes the humanization GFP gene that is suitable for expressing in mammalian cell.After transfection, select to express the cell of high-level GFP and use MoFlo cell counter (CytomatioN, INc.) sorting by facs analysis at first.Effective inhibitor Decitabine of Mammals DNMT1 is used as positive control.In order to screen the reactivate that is used for CMV-EE210, add Decitabine (1 μ M) or test compound (concentration is 30-50 μ M) to perfect medium (being supplemented with the DMEM that does not contain Phenol Red (Gibco, Life TechNologies) of 10% foetal calf serum (HycloNe)).Then cell is inoculated in comprising 96 orifice plates of test compound is 30% to converge (～5000 cells/well) and at 5% CO ₂In 37 ℃ the growth three days.Plate is checked under fluorescent microscope, used 450-490 exciter filter (I3 filtercube, Leica, Deerfield IL).If 10% viable cell is expressed GFP, then the hole is evaluated as the g1 positive; If 30% viable cell is expressed GFP, then the hole is evaluated as the g2 positive; If express GFP with 75% viable cell, then the hole be evaluated as g3.GFPIC ₅₀Inhibitor concentration when making the dosage of GFP expression level from g3 to g1/2 is (as IC ₅₀).As shown in table 3, in the compound of test, six compounds (Compound D DD5, Compound D DD6, Compound D DD7, Compound D DD8, Compound D DD9 and Compound D DD10) recover the GFP gene transcription with the level greater than 75%.In addition, their toxicity is less than half of Decitabine.

Table 3:

Active and induce the summary of DNMT1 degraded about the demethylation of the The compounds of this invention selected

ND-does not detect activity; TD ₅₀The dosage of-50% cell survival; TBT-remains to be tested

The RT-PCR test of genetic expression

With the SN31439.Cl of RKO and HCT-116 cell usefulness different concns, SN31486.Cl and SN31575.CL handle.After 48 hours cultivation, harvested cell is used to use Qiagen Rneasy MiNi test kit to carry out RNA and separates.RNA uses spectrophotometer to quantize, and the cDNA that the RNA of 1 μ g is used to use Bio-Rad iScript cDNA Synthesis test kit to carry out is synthetic.The SYBER GreenER qPCRSuperMix that use is used for iCycler carries out RTPCR according to manufacturer's rules.The following p165 ' of primer sequence-atgtcctgccttttaacgta-3 ' and the 5 '-gtgctcactccagaaaactc-3 ' that uses, MLH-1 5 '-tgaggaagggaacctgattg-3 ' and 5 '-tcttcgtcccaattcacctc-3 ', p15 5 '-caccatgaagcgaaacacag-3 ' and 5 '-tccatcggaagattcgtagc-3 ', GAPDH 5 '-attgccctcaacgaccactt-3 ' and 5 '-ggtccaccaccctgttgc-3 ' and B-actin 5 '-ctggaacggtgaaggtgaca-3 ' and 5 '-aagggacttcctgtaacaacgca-3 '.Sample is analyzed deriving from the iQ5 Multicolor PCR in real time detection system of Bio-Rad.Use iQ5 Optical system software release 2.0 to carry out data analysis, and (Threshold Cycle is CT) with CT mean value to measure the threshold cycle from this software.For each sample, carry out four reactions at least, two are used for p16, and two are used for house keeper's GAPDH contrast in addition.Deduct the mean CT-number that derives from the GAPDH reaction from each corresponding p16 sample CT, obtain being called the numerical value of Δ CT.Then, deduct the Δ CT that derives from undressed p16 sample, obtain being called another numerical value of Δ Δ CT from all treated Δ CT values.Relative expression's value equals the power power (=2^-Δ Δ CT) of negative Δ Δ CT value 2.The duplicate sample is averaged together, obtain the average relative expression of each processing, and base of calculation is poor.Use identical method to calculate relative expression's value of p15 and MLH-1.

As shown in fig. 1, compd A AA, compd B BB and Compound C CC can increase the rna expression level of p16.Concentration is that the SN31439 of 0.1 μ M causes that the p16 that is better than undressed maximum expresses again.Because higher levels of p16 is seemingly causing that p16 has less effect aspect the expression again, so have about drug solubility or more toxic problems at the higher concentration of test.

Growth test based on cell

Test based on cell cultures can be used for estimating the ability that The compounds of this invention suppresses one or more cytoactives (for example, growth of cancer cells and/or survival).Many cancerous cell lines can derive from American Type Culture Collection, and (American Type Culture Collection is ATCC) with other source.In brief, with cell inoculation opaque and white 96 orifice plates of handling through tissue culture (Thermo Electron, Vantaa is Finland) in the suitable growth medium in (being determined by ATCC), the speed that depends on cell proliferation, every hole are 5000-10000 cell.Make cellular exposure growth 96 hours in the presence of the medicine of proper concn or the DMSO of equivalent (pharmaceutical diluents) and the DMSO that allows in the medicine of proper concn or equivalent then.After this, to each hole add 100 μ l Cell-Titer-Glo (CTG) reagent (Promega, Inc., Madison, WI).Then plate is at room temperature shaken 2 minutes allowing cytolysis, and at room temperature incubation 10 minutes so that the fluorescent signal stabilization.Similar to the Kinase-Glo test reagent that derives from Promega, this reagent comprises luciferase and substrate fluorescein thereof simultaneously.By the conversion of the ATP activated luciferase catalysis fluorescein in the cellular lysate to oxyluciferin, this is a reaction that produces light.The amount of the light that produces is directly proportional with the amount of ATP in the cellular lysate, and the amount of described ATP itself is directly proportional with cell count and provides the index of cell proliferation.The C of representative compounds ₅₀Value is provided in the following table 4.

Table 4:

Although described the present invention with reference to some embodiment and embodiment, should be appreciated that, can further modify and change and do not break away from spirit of the present invention or scope embodiment and embodiment.

Claims (35)

1. the compound of formula (I) or its physiologically acceptable salt or phosphate precursor medicine or carboxylicesters or amino acid ester prodrug,

G wherein ₁, G ₂, G ₃, and G ₄Be that C, N or N+ are (at R independently of one another ⁶-R ⁹When being connected in N); G ₅And G ₆Be CH or N independently of one another; G ₇And G ₈Be that CH, C are (at R independently of one another ²When being connected in C), N or N+ be (at R ²When being connected in N),

D ₁And D ₂Be respectively CH, C separately (at R ³When being connected in C), N or N+ be (at R ³When being connected in N),

R ⁶, R ⁷, R ⁸, and R ⁹Be respectively H, halogen, CF separately ₃, OCF ₃, CN, CONHR ⁴, CONR ⁴R ⁵, SO ₂Me, SO ₂NHR ⁴, SO ₂NR ⁴R ⁵, NHCOR ⁴, NHR ⁴, NR ⁴R ⁵, OR ⁴, NO ₂, or CH ₂R ⁴, R wherein ⁴And R ⁵Be H, rudimentary C independently of one another ₁-C ₆Alkyl or cycloalkyl, described rudimentary C ₁-C ₆Alkyl or cycloalkyl is randomly replaced by amino, hydroxyl or methoxy group, or has one or more Sauerstoffatoms or nitrogen-atoms a part as this cycloalkyl structure, this cycloalkyl can be represented morpholine, tetramethyleneimine, piperidines, imidazoles or 4-methylpiperazine, perhaps can encircle carbon by-N=replacement-CH=

R ²And R ³Be H, NHR independently of one another ⁴, NR ⁴R ⁵, OR ⁴, NO ₂Or CH ₂R ⁴, R wherein ⁴And R ⁵It is as defined above,

X can be H or C ₁-C ₆Alkyl, described C ₁-C ₆Alkyl is randomly replaced by amino, hydroxyl or methoxy group, or has one or more Sauerstoffatoms or the nitrogen-atoms part as the cycloalkyl structure, and it can represent azetidine, tetramethyleneimine, piperidines, piperazine or morpholine;

Y can be CONR ⁴, NR ⁴CO, O, S (O) _n[N=0-2], (CH ₂) _k[k=1-6] ,-CH=CH-, NR ⁴, or the direct connection between two aromatic nucleus (that is, the C-C key between two aromatic nucleus), wherein R ⁴And R ⁵As defined above;

O, m and p represent the link position of group Z;

Z can be one of group Q1-Q43 shown in the formula (II);

Wherein A is O or NR ⁴, R wherein ⁴It is as defined above,

G ₉-G ₁₃Be that C, N or N+ are (at R independently of one another ¹⁰-R ¹³When being connected in N); But G ₉-G ₁₃In at least three be C; With

R ¹⁰, R ¹¹, R ¹², R ¹³, and R ¹⁴Be respectively H, halogen, alkyl, CF separately ₃, OCF ₃, CN, CONHR ⁴, CONR ⁴R ⁵, SO ₂Me, SO ₂NHR ⁴, SO ₂NR ⁴R ⁵, NHCOR ⁴, NHR ⁴, NR ⁴R ⁵, OR ⁴, NO ₂Or CH ₂R ⁴, R wherein ⁴And R ⁵As defined above.

2. the compound of formula (III),

Or its pharmacologically acceptable salt, phosphate precursor medicine or carboxylicesters or amino acid ester prodrug, wherein

R ⁶, R ⁷, R ⁸, and R ⁹Be respectively H, halogen, CF separately ₃, OCF ₃, CN, CONHR ⁴, CONR ⁴R ⁵, SO ₂Me, SO ₂NHR ⁴, SO ₂NR ⁴R ⁵, NHCOR ⁴, NHR ⁴, NR ⁴R ⁵, OR ⁴, NO ₂Or CH ₂R ⁴, R wherein ⁴And R ⁵Be H, C independently of one another ₁-C ₆Alkyl or cycloalkyl, described alkyl and cycloalkyl are randomly replaced by one or more amino, hydroxyl or methoxy group;

X is H or C ₁-C ₆Alkyl, described C ₁-C ₆Alkyl is randomly replaced by one or more amino, hydroxyl or methoxy group;

Y is CONR ⁴, NR ⁴CO, O, S (O) _n(n=0-2), (CH ₂) _k(k=1-6) ,-CH=CH-or NR ⁴, R wherein ⁴And R ⁵As defined above;

G ₇And G ₈Be CH or N independently of one another,

D ₁And D ₂Be CH or N independently of one another,

O, m and p represent the link position of group Z;

Z is one of group Q1-Q43 shown in the above-mentioned formula (II), wherein,

A is O or NR ⁴, R wherein ⁴As defined above;

G ₉-G ₁₃Be that C, N or N+ are (at R independently of one another ¹⁰-R ¹³When being connected in N), but G ₉-G ₁At least three in 3 is C,

R ¹⁰, R ¹¹, R ¹², R ¹³, and R ¹⁴Be respectively H, halogen, alkyl, CF separately ₃, OCF ₃, CN, CONHR ⁴, CONR ⁴R ⁵, SO ₂Me, SO ₂NHR ⁴, SO ₂NR ⁴R ⁵, NHCOR ⁴, NHR ⁴, NR ⁴R ⁵, OR ⁴, NO ₂Or CH ₂R ⁴, and R wherein ⁴And R ⁵As defined above.

3. the salt of the compound of claim 1, wherein said salt is to form with following acid: hydrochloric acid, Hydrogen bromide, sulfuric acid, phosphoric acid, carboxylic acid, sulfonic acid, thioic acid sulfoacid or phosphonic acids, acetate, propionic acid, oxyacetic acid, Succinic Acid, toxilic acid, hydroxymaleic acid, methyl-maleic acid, FUMARIC ACID TECH GRADE, oxysuccinic acid, tartrate, lactic acid, oxalic acid, glyconic acid, saccharic acid, glucuronic acid, citric acid, phenylformic acid, styracin, amygdalic acid, Whitfield's ointment, the 4-aminosallcylic acid, the 2-phenoxy benzoic acid, the 2-acetoxy-benzoic acid, pamoic acid, nicotinic acid, Yi Yansuan, amino acid, L-glutamic acid, Aspartic Acid, phenylacetic acid, methylsulfonic acid, ethyl sulfonic acid, the 2-hydroxyethanesulfonic acid, ethane-1, the 2-disulfonic acid, Phenylsulfonic acid, the 4-toluene sulfonic acide, naphthalene-2-sulfonic acid, naphthalene-1, the 5-disulfonic acid, 2-or 3-phoshoglyceric acid, Robison ester, N-cyclohexyl thionamic acid, and xitix.

4. the salt of the compound of claim 1, wherein said salt is sodium salt, calcium salt, lithium salts, sylvite, ammonium salt or trialkyl ammonium salts.

5. the compound of claim 1, wherein Z is Q1, Q2, Q3, Q4, Q9 or Q10.

6. the compound of claim 1, wherein R ⁶, R ⁷, R ⁸, and R ⁹H respectively does for oneself; X is H; Y is CONH or NHCO; Z is Q2 or Q9; A is that NH and Z are connected m or p position.

7. the compound of claim 1, wherein R ⁶, R ⁷, R ⁸, and R ⁹H respectively does for oneself; X is H; Y is that CONH, Z are Q2; A is NH; Be connected the p position with Z.

8. the compound of claim 1, wherein R ⁶, R ⁷, R ⁸, and R ⁹H respectively does for oneself; X is H; Y is CONH; Z is Q9; A is NH; Be connected the p position with Z.

9. the compound of claim 1, wherein R ⁶, R ⁷, R ⁸, and R ⁹H respectively does for oneself; X is H; Y is CONH or NHCO; Z is Q2; A is that NH and Z are connected the m position.

10. the compound of claim 1, wherein R ⁶, R ⁸, and R ⁹H respectively does for oneself; R ⁷Be NMe ₂X is H; Y is CONH or NHCO; Z is Q2; A is that NH and Z are connected the m position.

11. the compound of claim 1, wherein R ⁶, R ⁸, and R ⁹H respectively does for oneself; R ⁷Be Cl; X is H; Y is CONH; Z is Q2; A is that NH and Z are connected the p position.

12. the compound of claim 1, wherein R ⁶, R ⁷, R ⁸, and R ⁹Be respectively H, F or Cl.

13. pharmaceutical composition, it comprises compound, salt or the prodrug of claim 1, and pharmaceutically acceptable carrier.

14. the pharmaceutical composition of claim 13, wherein said compound, salt or prodrug are solid form.

15. the pharmaceutical composition of claim 13, wherein said pharmaceutical composition are oral dosage form.

16. the pharmaceutical composition of claim 13, wherein said pharmaceutical composition are injectable formulation.

17. the pharmaceutical composition of claim 13, wherein said pharmaceutical composition are topical dosage forms.

18. the method for the dna methylation in the inhibition cell comprises: make compound, salt or the prodrug of cells contacting claim 1, make that the dna methylation activity in the cell is suppressed.

19. the method for the dna methylation in the inhibition cell comprises: make compound, salt or the prodrug of cells contacting claim 1, make that the dnmt rna activity in the cell is suppressed.

20. the method for claim 19, the activity of wherein said dnmt rna is suppressed by the degraded of dnmt rna DNMT1.

21. the method for claim 19, the step of wherein said contact comprise compound, salt or the prodrug of the claim 1 that makes cells contacting biology significant quantity, make dnmt rna DNMT1 activity inhibited at least 50% in the cell.

22. the method for claim 19, the step of wherein said contact comprise compound, salt or the prodrug of the claim 1 that makes cells contacting biology significant quantity, make the activity of the dnmt rna DNMT1 in the cell suppress by at least 25%.

23. be used for recovering the active method of the repressed gene of dna methylation of cell, comprise: make compound, salt or the prodrug of the claim 1 of cells contacting biology significant quantity, make the activity of the repressed gene of dna methylation raise at least 25% with respect to the situation that does not have described compound, salt or prodrug to exist.

24. the method for claim 23, the step of wherein said contact comprises compound, salt or the prodrug of the claim 1 that makes cells contacting biology significant quantity, makes the transcriptional activity of the gene transcription thing that dna methylation is suppressed or transcriptional level raise at least 25%.

25. the method for claim 23, the repressed gene of wherein said dna methylation is selected from following group: 14-3-3Sigma, ABL1 (P1), ABO, APC, AR (androgen receptor), BLT1 (leukotrienes B4 acceptor), BRCA1, CALCA (thyrocalcitonin), CASP8 (caspase 8), caveolin 1, CD44, CFTR, COX2, CSPG2 (versican), CX26 (connection protein 26), Cyclin A1, DBCCR1, ECAD (E-cadherin), endothelin-receptor B, EPHA3, EPO (erythropoietin), ER (estrogen receptor), FHIT, GPC3 (glypican 3), GST-pi, H19, H-cadherin (CDH13), gamma globulin, HIC1, hMLH1, HOXA5, IGF2 (insulin-like growth factor II), IGFBP7, IRF7, LKB1, LRP-2 (huge albumen), MDGI (growth inhibiting factor in breast source), MDR1, MDR3 (PGY3), MGMT (06 methyl guanine methyl transferase), MUC2, MYOD1, N33, NEP (neutral endopeptidase 24.1)/CALLA, NIS (sodium iodide symport gene), P14/ARF, P15 (CDKN2B), P16 (CDKN2A), P27KIP1, p57KIP2, PAX6, PgR (PgR), RAR-β 2, RASSF1, RB1 (retinoblastoma), TERT, TESTIN, TGFBRI, THBS1 (thrombospondin-1), TIMP3, TLS3 (T-plastin), urokinase (uPA), VHL (Hippel gene), WT1, and ZO2 (zonuls occludens 2).

26. be used for the treatment of the method for suffering from the patient of unusual dna methylation diseases associated, comprise: give pharmaceutical composition described patient, described pharmaceutical composition comprises compound, salt or the prodrug of the claim 1 for the treatment of significant quantity, and pharmaceutically acceptable carrier.

27. the method for claim 26, wherein said pharmaceutical composition are by following administration: oral administration, parenterai administration, topical, intraperitoneal administration, intravenous administration, intra-arterial administration, transdermal administration, sublingual administration, intramuscular administration, rectal administration, transbuccally administration, intranasal administration, liposome administration, by inhalation, vagina administration, eye drops, by administration in local delivery administration, subcutaneous administration, the fat, intra-articular administration or intrathecal drug delivery.

28. the method for claim 26, wherein said drug composition oral administration.

29. the method for claim 26, it comprises in addition:

Give second therapeutical agent with the associating of described pharmaceutical composition to described patient.

30. the method for claim 29, wherein said second therapeutical agent is Decitabine or azacitidine.

31. the method for claim 29, wherein said second therapeutical agent are selected from medicine, biology medicine, interleukin-, Interferon, rabbit, cytokine, immunomodulator and the monoclonal antibody of histone deacylase inhibitor, antibiolics, alkylating agent, retinoid, hormone drug, plant origin.

32. the method for claim 31, wherein said histone deacylase inhibitor are selected from the two hydroxamic acid of bent ancient rhzomorph A, cork anilid hydroxamic acid, oxamflatin, cork acid, the two hydroxamic acid of a carboxylic acid styracin, pyroxamide, trapoxin A, apicidin, depsipeptide, N-(2-aminophenyl)-4-[N-(pyridin-3-yl methoxycarbonyl) amino methyl] benzamide, butyric acid, phenyl butyrate and arginine butyric ester.

33. the method for claim 26, wherein said and unusual dna methylation diseases associated is selected from hematology illness, innocent tumour and cancer.

34. the method for claim 33, wherein said hematology illness is selected from acute myelogenous leukemia, acute promyelocytic leukemia, acute lymphoblastic leukemia, chronic granulocytic leukemia, myelodysplastic syndrome and sicklemia disease.

35. the method for claim 33, wherein said cancer is selected from following group: mammary cancer, skin carcinoma, osteocarcinoma, prostate cancer, liver cancer, lung cancer, nonsmall-cell lung cancer, the cancer of the brain, laryngocarcinoma, carcinoma of gallbladder, carcinoma of the pancreas, the rectum cancer, parathyroid carcinoma, thyroid carcinoma, adrenal carcinoma, nervous center is organized cancer, head and neck cancer, colorectal carcinoma, cancer of the stomach, bronchogenic carcinoma, and kidney, rodent cancer, ulcer type and nipple type squamous cell carcinoma, the transitivity skin carcinoma, osteosarcoma, Ewing sarcoma, non-Hodgkin lymphomas (veticulum cell sarcoma), myelomatosis, giant cell tumor, small cell lung tumor, gallbladdergallstonecholetithiasis, islet cell tumor, primary brain tumors, acute and chronic lymphocytic knurl and granulocyte knurl, the hair cell knurl, adenoma, hyperplasia, medullary carcinoma, pheochromocytoma, mucous membrane neuroma (mucosal Neuronms), enteron aisle ganglioneuroma (intestinalganglloneuromas), hyperplasia corneal nerve knurl, marfanoid habitus knurl, Wilm ' s knurl, spermocytoma, ovarian tumor, leiomyoma, the cervical atypism hyperplasia, and carcinoma in situ, neuroblastoma, retinoblastoma, soft tissue sarcoma, the carcinoid malignant knurl, partial skin lesion, cutaneous T cell lymphoma (mycosis fungoide), rhabdosarcoma, Kaposi, osteogenic sarcoma, pernicious hypercalcemia, the nephrocyte tumour, polycythemia vera (polycythermia vera), gland cancer, diversity glioblastoma multiforme (glioblastomamultiforma), leukemia, lymphoma, malignant melanoma, and epidermoid carcinoma.

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