CN103539731A - Pyridine-2-amide compound as well as preparation method, pharmaceutical composition and use thereof - Google Patents
Pyridine-2-amide compound as well as preparation method, pharmaceutical composition and use thereof Download PDFInfo
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D405/00—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom
- C07D405/02—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom containing two hetero rings
- C07D405/12—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom containing two hetero rings linked by a chain containing hetero atoms as chain links
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- C07D213/00—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members
- C07D213/02—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members having three double bonds between ring members or between ring members and non-ring members
- C07D213/04—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members having three double bonds between ring members or between ring members and non-ring members having no bond between the ring nitrogen atom and a non-ring member or having only hydrogen or carbon atoms directly attached to the ring nitrogen atom
- C07D213/60—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members having three double bonds between ring members or between ring members and non-ring members having no bond between the ring nitrogen atom and a non-ring member or having only hydrogen or carbon atoms directly attached to the ring nitrogen atom with hetero atoms or with carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, e.g. ester or nitrile radicals, directly attached to ring carbon atoms
- C07D213/78—Carbon atoms having three bonds to hetero atoms, with at the most one bond to halogen, e.g. ester or nitrile radicals
- C07D213/81—Amides; Imides
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- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D401/00—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom
- C07D401/02—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom containing two hetero rings
- C07D401/12—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom containing two hetero rings linked by a chain containing hetero atoms as chain links
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- C07D409/00—Heterocyclic compounds containing two or more hetero rings, at least one ring having sulfur atoms as the only ring hetero atoms
- C07D409/02—Heterocyclic compounds containing two or more hetero rings, at least one ring having sulfur atoms as the only ring hetero atoms containing two hetero rings
- C07D409/12—Heterocyclic compounds containing two or more hetero rings, at least one ring having sulfur atoms as the only ring hetero atoms containing two hetero rings linked by a chain containing hetero atoms as chain links
Abstract
The invention relates to a pyridine-2-amide compound shown as a general formula I as well as a preparation method, a pharmaceutical composition and use thereof. The compound as an integrase inhibitor is used for preparing medicines for treating diseases mediated by various vetrovirus integrases including HIV-1 (Human Immunodeficiency Virus-1) integrase.
Description
Technical field
The present invention relates to medical technical field, be specifically related to pyridine-2-amides and preparation method thereof, containing pharmaceutical composition and the purposes of this compounds.
Background technology
HIV virus is retrovirus, and it invades human body cell need to pass through these important processes of absorption, fusion, reverse transcription, integration, expression and assembling.Wherein, integration is the process being connected with host cell DNA by integrating enzymatic viral DNA, intergrase is the necessary enzyme of HIV-1 virus replication, and human body cell does not have the corresponding component of function with it, and this becomes one of desirable target spot of anti-HIV-1 medicines.The listing of the integrase inhibitor Raltegravir of Merk company in 2007 has also proved that HIV-1 intergrase is an effective medicine target.
HIV-l intergrase (HIV-1IN) is that its functional structure is mainly divided into three regions: N petiolarea, catalytic core district, C petiolarea by 3 ' end pol genes encoding of virus, altogether containing 288 amino acid whose protein.Two reactions of the main catalysis of this enzyme: 3 ' tip cut-off reaction and chain transfer reaction of viral DNA.It first respectively cuts away two Nucleotide at the LTR 3 '-end of viral DNA specifically, make its expose after 3 '-CA end with viral DNA form integrate before mixture (PIC), displacement enters nucleus.In nucleus, intergrase cuts at random host cell DNA again and produces a stagger, then 3 ' damaged end of viral DNA is connected through transesterification reaction with 5 ' end of host DNA.
In integration process, divalent-metal ion is as Mg
2+, Mn
2+it is the necessary cofactor of intergrase performance catalysis.Up to the present, maximum method of research is by designing the compound that a class can chelated metal ions, with intergrase chelated metal ions competitively, makes its loss of catalytic activity, thereby reaches the object of inhibition.In addition, because HIV virus has variability, for fear of cross resistance, investigators also, striving to find site new on intergrase, mainly comprise the multimerization that suppresses intergrase, suppress that intergrase is combined with host cell cofactor etc.Compound in the present invention relates to these two inhibition sites on intergrase, below this is introduced respectively.
(1) active state when hiv integrase participates in catalysis 3 '-P process is dimer, active state while participating in catalysis ST process is the tetramer, therefore, can design a compounds, suppress intergrase and form polymer, thereby reach the object that suppresses its catalytic activity.
(2) LEDGF/p75 (lens epithelium-derived growth factor p75) is crystalline epidermis derivative growth factor p75 or is called that to transcribe the sub-p75 of co-activating (transcriptional co-activator p75) be the albumen of a 60kDa in host cell.Mutating experiment and LEDGF/p75 knock out the proofs such as experiment, and it has brought into play important effect in the integration process of HIV-1 virus.Research finds, is combined with viral integrase enzyme CCD in the C end IBD position of LEDGF/p75, and N holds PWWP position to combine with host cell chromosome, thereby between viral integrase enzyme and host cell, plays a function served as bridge, the completing of mediated integration process.Owing to there is the mechanism of action different from existing medicine, in recent years, block the more and more noticeable new target drone that is combined into of IN-LEDGF/p75.
Compound in the present invention relates to above two binding sites, and it is active that some compound has multiple inhibition simultaneously, is the integrase inhibitor of many target spots.Antiviral activity experiment showed, that some compounds that can be combined, show in vitro HIV-1 integrase inhibiting activities with these sites of intergrase can effectively suppress the infringement of virus to host cell.In sum, article pyridine compounds and their of the present invention and preparation method thereof and in the application that suppresses HIV-1 virus field.
Summary of the invention
The inventor carries out deep research to having the compound of HIV-1 integrase inhibiting activities, has designed and synthesized and has had the compound shown in formula I.Test result shows, these compounds have restraining effect to a plurality of sites of HIV-1 intergrase, can be used as HIV-1 integrase inhibitor, and may develop into anti-AIDS drug, have completed on this basis invention.
Therefore, the object of this invention is to provide pyridine-2-amides or its pharmacy acceptable salt as shown in general structure I.
Another object of the present invention is to provide the preparation method of above-claimed cpd.
Another object of the present invention is to provide above-claimed cpd or its pharmacy acceptable salt as the purposes of integrase inhibitor, more specifically, provide above-claimed cpd or its pharmacy acceptable salt as integrase inhibitor application in the medicine of the integrase mediated disease of the multiple retroviral that comprises HIV-1 intergrase in preparation treatment.The integrase mediated disease of multiple retroviral of the described HIV-1 of comprising intergrase comprises acquired immune deficiency syndrome (AIDS), hepatitis B, the third liver etc.
An also object of the present invention is to provide a kind of pyridine-2-amides shown in one or more formula I for the treatment of significant quantity or pharmaceutical composition of its pharmacy acceptable salt of comprising.
In a first aspect of the present invention, provide a class by the pyridine-2-amides shown in formula I or its pharmacy acceptable salt:
In formula I, L is: (1) key; (2) C
1-C
6alkylidene group; (3) C
2-C
6unsaturated alkylene; (4) (C
0-C
6alkylidene group)-(C
3-C
6cycloalkylidene)-(C
0-C
6alkylidene group); Or (5) (C
0-C
6alkylidene group)-M-(C
0-C
6alkylidene group), wherein, M is N (R
a), OC (=O) or C (=O) O; Wherein, the alkylidene group in the unsaturated alkylene in (3) and (2), (4), (5) not necessarily by 1-3 separately independently substituting group replace, described substituting group is selected among following groups: C
1-C
6alkyl, C
1-C
6alkoxy C
1-C
6alkyl, C
3-C
8cycloalkyl, halogen, sulfydryl, hydroxyl ,-CF
3,-CN ,-NO
2,-NR
ar
b,-NR
acOR
b,-NR
acOOR
b,-NR
asO
2r
b,-COOR
b,-COR
b,-CONR
ar
b,-SO
2r
b,-SO
2nR
ar
b,-OR
awith-OCOR
b;
R
1, R
2be hydrogen independently of one another; Hydroxyl; Or do not replace or by 1-3 the following groups that independently substituting group replaces separately: C
1-C
8alkyl, C
1-C
8alkoxy C
1-C
8alkyl, C
1-C
8alkoxyl group, C
2-C
8unsaturated alkyl, C
3-C
8cycloalkyl, C
6-C
12aryl, C
4-C
10heteroaryl or C
3-C
10heterocyclic radical; Described heteroaryl or heterocyclic radical comprise that 1-3 is selected from the heteroatoms in N, O and S; Described substituting group is selected from following groups: C
1-C
6alkyl, C
1-C
6alkoxyl group, C
1-C
6alkoxy C
1-C
6alkyl, halogen, amino, nitro, sulfydryl, hydroxyl ,-CN and-CF
3;
R
3for hydrogen, halogen, hydroxyl, sulfydryl ,-CF
3,-CN ,-NO
2, or do not replace or independently of one another by the following groups of 1-3 substituting group replacement: C
1-C
8alkyl, C
1-C
8alkoxyl group, C
2-C
8thiazolinyl, C
2-C
8alkynyl, C
3-C
8cycloalkyl, C
3-C
8cycloalkyloxy or amino; Described substituting group is selected among following groups: C
1-C
6alkyl, C
1-C
6alkoxyl group, C
1-C
6alkoxy C
1-C
6alkyl, halogen, sulfydryl, hydroxyl ,-CF
3,-CN ,-NO
2, ternary ,-NR amino to six-ring
ar
b,-NR
acOR
b,-NR
acOOR
b,-NR
asO
2r
b,-COOR
b,-COR
b,-CONR
ar
b,-SO
2r
b,-SO
2nR
ar
b,-OR
awith-OCOR
b; And R
3position be 3-position, 4-position, 5-position or the 6-position on pyridine ring;
Wherein, R
aand R
bbe hydrogen or C independently of one another
1-C
6alkyl;
Described halogen is fluorine, chlorine, bromine or iodine.
Preferably, in formula I,
L is C
1-C
4alkylidene group, C
2-C
4unsaturated alkylene;
R
1for hydrogen, C
1-C
8the alkyl of straight or branched, C
2-C
8the unsaturated alkyl of straight or branched;
R
2for hydrogen, C
1-C
8the alkyl of straight or branched, C
2-C
8the unsaturated alkyl of straight or branched, C
3-C
8cycloalkyl, do not replace or by 1-3 the C that independently substituting group replaces separately
6-C
12aryl, do not replace or by 1-3 the C that independently substituting group replaces separately
4-C
10heteroaryl, do not replace or by 1-3 the C that independently substituting group replaces separately
3-C
10heterocyclic radical, described heteroaryl or heterocyclic radical comprise 1-3 and are selected from the heteroatoms in N, O and S; Wherein, described substituting group is halogen, amino, nitro, hydroxyl, cyano group ,-CF
3, C
1-C
6alkoxyl group, C
1-C
6alkyl;
R
3for hydrogen, halogen, hydroxyl, sulfydryl ,-CF
3,-CN ,-NO
2, C
1-C
6alkyl, do not replace or by C
1-C
4the C that alkoxyl group replaces
1-C
6alkoxyl group.
Preferably, in formula I,
L is C
1-C
2alkylidene group;
R
1for hydrogen, C
1-C
6the alkyl of straight or branched, C
2-C
6the thiazolinyl of straight or branched;
R
2for hydrogen, the alkyl of C1-C6 straight or branched, C
3-C
6cycloalkyl, do not replace or by 1-3 the C that independently substituting group replaces separately
6-C
10aryl, do not replace or by 1-3 the C that independently substituting group replaces separately
4-C
9heteroaryl, do not replace or by 1-3 the C that independently substituting group replaces separately
3-C
8heterocyclic radical, described heteroaryl or heterocyclic radical comprise 1-3 and are selected from the heteroatoms in N, O and S; Wherein, described substituting group is halogen, amino, nitro, hydroxyl, cyano group ,-CF
3, C
1-C
4alkoxyl group, C
1-C
4alkyl;
R
3for hydrogen, halogen, hydroxyl, sulfydryl ,-CF
3,-CN ,-NO
2, C
1-C
4alkyl, do not replace or by C
1-C
2the C that alkoxyl group replaces
1-C
4alkoxyl group.
Preferably, in formula I,
L is methylene radical;
R
1for hydrogen, normal-butyl, isobutyl-, isopentyl, 2-methacrylic;
R
2for the alkyl of hydrogen, C1-C4 straight or branched, cyclohexyl, do not replace or by 1-3 phenyl, naphthyl, furyl, thienyl, the indyl that independently substituting group replaces separately; Wherein, described substituting group is halogen, amino, nitro, hydroxyl, cyano group ,-CF
3, methoxyl group;
R
3for hydrogen, halogen, hydroxyl, methoxyl group, methoxymethoxy.
Preferably, generalformulaⅰcompound is following compounds:
According to a further aspect in the invention, the invention provides the preparation method of the pyridine-2-amides shown in described general formula I, it is characterized in that: compound of Formula I is prepared by following methods:
Work as R
1during for H,
(a) compound 7-1 and amine
carry out condensation, obtain compound of Formula I;
Work as R
1while being not H,
(b) compound 7-1 and amine R
1nH
2carry out condensation, obtain compound 7-2;
(c) compound 7-2 and halohydrocarbon
there is nucleophilic reaction and obtain general formula compound I;
Wherein, X is fluorine, chlorine, bromine or iodine;
R
1, R
2, R
3identical with the definition in above-mentioned general formula I with the definition of L.
Reaction reagent and condition:
Work as R
1during for H, step is as follows: (a) compound 7-1 is dissolved in DMF, take EDCI(1-ethyl-3-(3-dimethylamine propyl) carbodiimide hydrochloride), HOBt(1-hydroxybenzotriazole) be condensing agent, DIPEA(N, N-diisopropylethylamine) make alkali, with corresponding amine
carry out condensation, under 50 ℃ of conditions of microwave, react and within 2 hours, obtain target compound I;
Work as R
1while being not H, step is as follows: (b) compound 7-1 is dissolved in DMF, take EDCI, HOBt to make alkali as condensing agent, DIPEA, first with chain aliphatic amide R
1nH
2carry out condensation, under 50 ℃ of conditions of microwave, react 2 hours, obtain midbody compound 7-2; (c) compound 7-2 is dissolved in DMF, with NaH, makes alkali, with halohydrocarbon
there is nucleophilic reaction and obtain corresponding target compound I.
In accordance with a further aspect of the present invention, be to provide above-claimed cpd or its pharmacy acceptable salt as integrase inhibitor, the purposes in preparation treatment in the medicine of the integrase mediated disease of the multiple retroviral that comprises HIV-1 intergrase.
The present invention also provides a kind of pharmaceutical composition as HIV-1 integrase inhibitor, and it contains the pyridine-2-amides shown in one or more formula I for the treatment of significant quantity or its pharmacy acceptable salt and pharmaceutically acceptable carrier.
Embodiment
Below in conjunction with specific embodiment, the invention will be further described, should be understood that these embodiment only do not limit the present invention for the present invention is described.
Preparation Example
Compound
1h-NMR spectroscopic data is measured and is used Varian Mercury-300MHz or Varian Mercury-400MHz nucleus magnetic resonance, ultimate analysis to use Vario EL determinator, and fusing point is measured with Buchi-510 capillary tube technique, and temperature is not calibrated.Infrared spectra is by Bio-Rad FTS-185 determination of infrared spectroscopy; Finnigan MAT 95 mass spectrographs for mass spectrum EI-MS, ESI-MS is used Finnigan LCQ Deca mass spectrograph to measure.Specific rotation is by P-1030(A012360639) automatic polarimeter mensuration.Rapid column chromatography carries out on silica gel H (10-40 μ M).Starting compound 7-1Cong AlfaAesar company buys; EDCI, HOBt, DIPEA buy from gill biochemical corp; All the other each reagent are not specifically noted and are business purchase, without being further purified, are directly used in reaction.Solvent purification is with reference to Purification of laboratory Chemicals; D.D.Perrin; W.L.F.Armarego and D.R.Perrin Eds., Pergamon Press:Oxiford, 1980.
Embodiment 1
N-(furans-2-methyl)-3-hydroxyl-2-pyridine carboxamide (LZ-21)
By compound 3-pyridone-2-carboxylic acid (100mg, 0.719mmol), HOBt (194mg, 1.438mmol), EDCI (276mg, 1.438mmol) be dissolved in 5ml DMF, add DIPEA (251 μ l, 1.438mmol), stirring at room 15 minutes, dropwise add 2-aminomethyl-furan (127 μ l, 1.438mmol).Put into microwave reactor (CEM, Discovery), 50 ℃ are reacted 2 hours.Be spin-dried for solvent, methylene dichloride dissolves, and washes 2 times, and saturated sodium-chloride is washed 1 time, anhydrous sodium sulfate drying.Column chromatography (petrol ether/ethyl acetate=12/1) obtains white solid compound L Z-21 (148mg, 94%).
1h NMR (300MHz, CDCl
3) δ 12.01 (s, 1H), 8.32 (s, 1H), 8.05 (d, J=4.0Hz, 1H), 7.39 (d, J=0.9Hz, 1H), 7.35-7.30 (m, 2H), 6.33 (d, J=5.9Hz, 2H), 4.64 (d, J=6.0Hz, 2H) .EI-MS m/z:218 (M)
+.
Embodiment 2
N-(thiophene-2-methyl)-3-hydroxyl-2-pyridine carboxamide (LZ-22)
Synthetic method, with compound LZ-21, just changes raw material 2-aminomethyl-furan into 2-aminomethyl thiophene.Obtain white solid compound L Z-22 (152mg, 90%).
1h NMR (300MHz, CDCl
3) δ 12.01 (s, 1H), 8.34 (s, 1H), 8.04 (dd, J=3.9,1.8Hz, 1H), 7.33 (t, J=3.0Hz, 2H), 7.25 (d, J=1.2Hz, 1H), 7.06 (d, J=2.6Hz, 1H), 7.01-6.95 (m, 1H), 4.81 (d, J=6.1Hz, 2H) .EI-MS m/z:234 (M)
+.
Embodiment 3
N-(cyclohexyl methyl)-3-hydroxyl-2-pyridine carboxamide (LZ-25)
Synthetic method, with compound LZ-21, just changes raw material 2-aminomethyl-furan into cyclohexyl methylamine.Obtain white solid compound L Z-25 (147mg, 87%).
1h NMR (300MHz, CDCl
3) δ 12.27 (s, 1H), 8.04 (dd, J=4.0,1.8Hz, 1H), 7.34-7.27 (m, 2H), 4.83 (s, 1H), 3.29 (t, J=6.6Hz, 2H), 1.76 (dd, J=27.3,12.4Hz, 5H), 1.23 (dd, J=21.0,9.6Hz, 4H), 1.02 (dd, J=23.3,10.6Hz, 2H) .EI-MS m/z:234 (M)
+.
Embodiment 4
N-(the fluoro-benzyl of 4-)-3-hydroxyl-2-pyridine carboxamide (LZ-26)
Synthetic method, with compound LZ-21, just changes raw material 2-aminomethyl-furan into 4-fluoro-benzylamine.Obtain white solid compound L Z-26 (166mg, 94%).
1h NMR (300MHz, CDCl
3) δ 12.05 (s, 1H), 8.33 (s, 1H), 8.04 (dd, J=3.9,1.8Hz, 1H), 7.38-7.27 (m, 4H), 7.04 (t, J=8.7Hz, 2H), 4.61 (d, J=6.2Hz, 2H) .EI-MS m/z:246 (M)
+.
Embodiment 5
N-(1-naphthyl-methyl)-3-hydroxyl-2-pyridine carboxamide (LZ-30)
Synthetic method, with compound LZ-21, just changes raw material 2-aminomethyl-furan into 1-aminomethyl naphthalene.Obtain white solid compound L Z-30 (163mg, 82%).
1h NMR (300MHz, CDCl
3) δ 12.28 (s, 1H), 8.50 (s, 1H), 8.10 (d, J=8.5Hz, 1H), 8.02-7.97 (m, 1H), 7.87 (dd, J=15.2,8.3Hz, 2H), 7.52 (dq, J=15.1,6.9Hz, 4H), 7.35 (d, J=3.5Hz, 2H), 5.10 (d, J=5.9Hz, 2H) .EI-MS m/z:278 (M)
+.
Embodiment 6
N-(4-trifluoromethyl-benzyl)-3-hydroxyl-2-pyridine carboxamide (LZ-31)
Synthetic method, with compound LZ-21, just changes raw material 2-aminomethyl-furan into 4-trifluoromethyl-benzylamine.Obtain white solid compound L Z-31 (173mg, 81%).
1h NMR (300MHz, CDCl
3) δ 12.06 (s, 1H), 8.61 (s, 1H), 8.07 (dd, J=3.8,2.0Hz, 1H), 7.62 (d, J=8.1Hz, 2H), 7.49 (d, J=8.0Hz, 2H), 7.39 (dd, J=2.8,2.2Hz, 2H), 4.71 (d, J=6.3Hz, 2H).
Embodiment 7
N-(indoles-5-methyl)-3-hydroxyl-2-pyridine carboxamide (LZ-32)
Synthetic method, with compound LZ-21, just changes raw material 2-aminomethyl-furan into 5-aminomethyl indoles.Obtain pale pink solid chemical compound LZ-32 (161mg, 84%).
1h NMR (300MHz, CDCl
3) δ 7.95 (s, 1H), 7.53 (s, 1H), 7.42-7.19 (m, 4H), 7.15 (s, 1H), 7.09 (s, 1H), 4.62 (s, 2H) .EI-MS m/z:267 (M)
+.
Embodiment 8
N-(4-amino-benzyl)-3-hydroxyl-2-pyridine carboxamide (LZ-33)
Synthetic method, with compound LZ-21, just changes raw material 2-aminomethyl-furan into 4-amino-benzylamine.Obtain pale pink solid chemical compound LZ-33 (149mg, 85%).
1h NMR (300MHz, CD
3oD) δ 8.08 (dd, J=4.4,1.3Hz, 1H), 7.45-7.38 (m, 1H), 7.31 (d, J=8.5Hz, 1H), 7.16 (d, J=8.6Hz, 2H), 6.77 (d, J=8.5Hz, 2H), 4.46 (s, 2H) .EI-MS m/z:243 (M)
+.
Embodiment 9
N-(3,4,5-trimethoxy-benzyl)-3-hydroxyl-2-pyridine carboxamide (LZ-34)
Synthetic method, with compound LZ-21, just changes raw material 2-aminomethyl-furan into 3,4,5-trimethoxy benzylamine.Obtain white solid compound L Z-34 (192mg, 84%).
1h NMR (300MHz, CDCl
3) δ 12.54 (s, 1H), 8.99 (s, 1H), 8.09 (s, 1H), 7.50 (s, 2H), 6.62 (s, 2H), 4.58 (d, J=5.9Hz, 2H), 3.87 (s, 6H), 3.83 (s, 3H) .EI-MS m/z:318 (M)
+.
Embodiment 10
N-(the fluoro-benzyl of 4-)-3-methoxyl group-2-pyridine carboxamide (LZ-53)
Step (1): the preparation of 3-methoxyl group-2-pyridine carboxylic acid methyl esters
Compound 3-hydroxyl-2-pyridine carboxylic acid (350mg, 2.52mmol), salt of wormwood (871mg, 6.3mmol) are suspended in 25ml acetone, in stirring at room.Methyl iodide (345 μ l, 5.54mmol) is dropwise added in above-mentioned suspension, and reflux is spent the night.Be spin-dried for solvent, methylene dichloride dissolves, and washes twice, and saturated sodium-chloride is washed once, and column chromatography (petrol ether/ethyl acetate=1/2) obtains colourless oil liquid 3-methoxyl group-2-pyridine carboxylic acid methyl esters (261mg, 62%).
1H?NMR(300MHz,CDCl
3)δ8.29(d,J=4.4Hz,1H),7.42(dt,J=16.5,6.5Hz,2H),3.98(s,3H),3.93(s,3H).
Step (2): the preparation of 3-methoxyl group-2-pyridine carboxylic acid
Compound 3-methoxyl group-2-pyridine carboxylic acid methyl esters (261mg, 1.56mmol), sodium hydroxide (281mg, 7.03mmol) were dissolved in methyl alcohol (9ml) and water (6ml), in stirring at room 1.5 hours.Be spin-dried for solvent, be dissolved in water, ethyl acetate extraction 5 times, merges organic phase, is spin-dried for and directly casts single step reaction.
1H?NMR?(300MHz,CD
3OD)δ8.18(s,1H),7.78(d,J=8.3Hz,1H),7.67(s,1H),3.96(s,3H).
Step (3): N-(the fluoro-benzyl of 4-)-3-methoxyl group-2-pyridine carboxamide (LZ-53)
The synthetic method of compound L Z-53, with compound LZ-26, just changes raw material 3-hydroxyl-2-pyridine carboxylic acid into 3-methoxyl group-2-pyridine carboxylic acid, obtains brown oily compound L Z-53 (61mg, 81%).
1H?NMR(300MHz,CDCl
3)δ8.45(d,J=2.9Hz,1H),7.75(t,J=9.4Hz,2H),7.40-7.31(m,2H),6.99(t,J=8.6Hz,2H),4.62(d,J=5.6Hz,2H),4.08(s,3H).
Embodiment 11
N-(the fluoro-benzyl of 4-)-2-pyridine carboxamide (LZ-54)
Synthetic method, with compound LZ-26, just changes raw material 3-hydroxyl-2-pyridine carboxylic acid into 2-pyridine carboxylic acid.Obtain colourless oil liquid compound L Z-54 (311mg, 94%).
1h NMR (300MHz, CDCl
3) δ 8.53 (d, J=4.8Hz, 1H), 8.42 (s, 1H), 8.24 (d, J=7.8Hz, 1H), 7.88 (td, J=7.7,1.6Hz, 1H), 7.48-7.40 (m, 1H), 7.38-7.29 (m, 2H), (7.02 t, J=8.7Hz, 2H), 4.63 (d, J=6.2Hz, 2H).
Embodiment 12
N-(the fluoro-benzyl of 4-)-6-hydroxyl-2-pyridine carboxamide (LZ-55)
Synthetic method, with compound LZ-26, just changes raw material 3-hydroxyl-2-pyridine carboxylic acid into 6-hydroxyl-2-pyridine carboxylic acid.Obtain colourless oil liquid compound L Z-55 (321mg, 91%).
1h NMR (300MHz, CD
3oD) δ 7.69 (s, 1H), 7.40-7.31 (m, 2H), 7.17 (d, J=12.2Hz, 1H), 7.05 (t, J=8.7Hz, 2H), 6.75 (d, J=9.0Hz, 1H), 4.54 (s, 2H).
Embodiment 13
N-(the fluoro-benzyl of 4-)-5-hydroxyl-2-pyridine carboxamide (LZ-56)
Synthetic method, with compound LZ-26, just changes raw material 3-hydroxyl-2-pyridine carboxylic acid into 5-hydroxyl-2-pyridine carboxylic acid.Obtain colourless oil liquid compound L Z-56 (164mg, 93%).
1h NMR (300MHz, CD
3oD) δ 8.14 (d, J=2.8Hz, 1H), 7.95 (d, J=8.6Hz, 1H), 7.36 (dd, J=8.4,5.4Hz, 2H), 7.25 (dd, J=8.6,2.8Hz, 1H), 7.03 (t, J=8.8Hz, 2H), 4.55 (s, 2H).
Embodiment 14
N-(the fluoro-benzyl of 4-)-4-hydroxyl-2-pyridine carboxamide (LZ-57)
Synthetic method, with compound LZ-26, just changes raw material 3-hydroxyl-2-pyridine carboxylic acid into 4-hydroxyl-2-pyridine carboxylic acid.
1H?NMR(300MHz,CDCl
3)δ8.76(s,1H),8.35(d,J=5.9Hz,1H),7.83(d,J=2.6Hz,1H),7.40-7.31(m,2H),7.02(t,J=8.7Hz,3H),4.63(d,J=6.2Hz,2H.
Embodiment 15
N-normal-butyl-3-hydroxyl-2-pyridine carboxamide (LZ-58)
Synthetic method, with compound LZ-26, just changes raw material 4-flunamine into 1-n-Butyl Amine 99.Obtain colourless oil liquid compound L Z-58 (246mg, 88%).
Embodiment 16
N-(the fluoro-benzyl of 4-) the fluoro-2-pyridine carboxamide of-3-(LZ-59)
Synthetic method, with compound LZ-26, just changes raw material 3-hydroxyl-2-pyridine carboxylic acid into 3-fluoro-2-pyridine carboxylic acid.Obtain white solid compound L Z-59 (322mg, 90%).
1h NMR (300MHz, CDCl
3) δ 8.36 (d, J=4.4Hz, 1H), 8.17 (s, 1H), 7.62-7.52 (m, 1H), 7.48 (dt, J=8.3,4.0Hz, 1H), 7.35 (dd, J=8.5,5.4Hz, 2H), 7.03 (t, J=8.7Hz, 2H), 4.61 (d, J=6.1Hz, 2H) .EI-MS m/z:248 (M)
+.
Embodiment 17
N-(the fluoro-benzyl of 4-)-3-(O-MOM)-2-pyridine carboxamide (LZ-60)
The preparation of step (1): N-(the fluoro-benzyl of 4-)-3-hydroxyl-2-pyridine carboxamide
Synthetic method, with compound LZ-26, obtains white solid compound N-(the fluoro-benzyl of 4-)-3-hydroxyl-2-pyridine carboxamide (895mg, 92%).
1H?NMR(300MHz,CDCl
3)δ12.05(s,1H),8.33(s,1H),8.04(dd,J=3.9,1.8Hz,1H),7.38-7.27(m,4H),7.04(t,J=8.7Hz,2H),4.61(d,J=6.2Hz,2H).
The preparation of step (2): N-(the fluoro-benzyl of 4-)-3-(O-MOM)-2-pyridine carboxamide (LZ-60)
N-(the fluoro-benzyl of 4-)-3-hydroxyl-2-pyridine carboxamide (223mg, 0.906mmol) is dissolved in the methylene dichloride that 10ml is dry, adds DIPEA (238 μ l, 1.359mmol), nitrogen protection, ice bath to 0 ℃ stirring.MOMCl (83 μ l, 1.087mmol) is dissolved in the methylene dichloride that 1ml is dry, under ice bath, dropwise adds in reaction solution.Remove ice bath, be slowly warming up to stirring at room 36 hours.Be spin-dried for solvent, column chromatography (petrol ether/ethyl acetate=1/3) obtains white solid compound L Z-60 (192mg, 73%).
1H?NMR(300MHz,CDCl
3)δ8.23(d,J=4.6Hz,1H),8.10(s,1H),7.66(d,J=8.5Hz,1H),7.36(d,J=5.6Hz,3H),7.03(q,J=8.4Hz,2H),5.28(sz,2H),4.61(d,J=6.0Hz,2H),3.53(s,3H).
Embodiment 18
N-normal-butyl-N-(the fluoro-benzyl of 4-)-2-pyridine carboxamide (LZ-61)
Step (1): the preparation of N-normal-butyl-2-pyridine carboxamide
Synthetic method, with compound LZ-26, just changes raw material 3-hydroxyl-2-pyridine carboxylic acid into 2-pyridine carboxylic acid, changes NSC 158269 into n-Butyl Amine 99.Obtain faint yellow oily body compound N-normal-butyl-2-pyridine carboxamide (686mg, 95%).
Step (2): the preparation of N-normal-butyl-N-(the fluoro-benzyl of 4-)-2-pyridine carboxamide (LZ-61)
NaH (154mg, 3.852mmol) is placed in to reaction flask, vacuumizes nitrogen protection, ice bath to 0 ℃ stirring.By N-normal-butyl-2-pyridine carboxamide (343mg, 1.926mmol) be dissolved in 10ml dry DMF, dropwise add in NaH, ice bath stirs 30 minutes, remove ice bath, be warming up to stirring at room 1 hour, to fluorine bromobenzyl (360 μ l, 2.89mmol) dropwise add in reaction solution, in stirring at room 8 hours.Be spin-dried for solvent, methylene dichloride dissolves, and washes twice, and saturated sodium-chloride is washed once, and column chromatography (petrol ether/ethyl acetate=2/1) obtains colourless oil liquid compound L Z-61 (452mg, 82%).
1H?NMR(300MHz,CDCl
3)δ8.64(dd,J=10.5,4.6Hz,1H),7.95(dd,J=16.5,7.9Hz,1H),7.68(dd,J=13.1,7.9Hz,1H),7.54-7.45(m,1H),7.40(dd,J=8.4,5.4Hz,1H),7.23-7.13(m,1H),7.01(dt,J=20.7,8.7Hz,2H),4.71(d,J=31.6Hz,2H),3.54-3.41(m,1H),3.34-3.24(m,1H),1.67(dt,J=15.1,7.4Hz,1H),1.49(dt,J=15.0,7.4Hz,1H),1.36(dt,J=15.0,7.4Hz,1H),1.08(dq,J=14.6,7.3Hz,1H),0.83(dt,J=61.6,7.3Hz,3H).
Embodiment 19
N-normal-butyl-N-(the fluoro-benzyl of 4-)-3-hydroxyl-2-pyridine carboxamide (LZ-62)
Step (1): the preparation of N-normal-butyl-3-hydroxyl-2-pyridine carboxamide
Synthetic method, with compound LZ-26, just changes raw material NSC 158269 into n-Butyl Amine 99.Obtain faint yellow oily body compound N-normal-butyl-2-pyridine carboxamide (246mg, 88%).
Step (2): the preparation of N-normal-butyl-3-(O-MOM)-2-pyridine carboxamide
Synthetic method, with compound LZ-60, just changes raw material N-(the fluoro-benzyl of 4-)-3-hydroxyl-2-pyridine carboxamide into N-normal-butyl-3-hydroxyl-2-pyridine carboxamide.Obtain faint yellow oily body compound N-normal-butyl-3-(O-MOM)-2-pyridine carboxamide (220mg, 73%).
1H?NMR(300MHz,CD
3OD)δ8.21(d,J=4.7Hz,1H),7.71(d,J=8.5Hz,1H),7.45(dd,J=8.6,4.6Hz,1H),5.30(s,2H),3.48(d,J=0.8Hz,3H),3.40(t,J=7.0Hz,2H),1.67-1.55(m,2H),1.44(dd,J=14.7,7.5Hz,2H),0.97(t,J=7.2Hz,3H).
Step (3): the preparation of N-normal-butyl-N-(the fluoro-benzyl of 4-)-3-(O-MOM)-2-pyridine carboxamide
Synthetic method, with compound LZ-61, just changes raw material N-normal-butyl-2-pyridine carboxamide into N-normal-butyl-3-(O-MOM)-2-pyridine carboxamide.Obtain colorless oil compound N-normal-butyl-N-(the fluoro-benzyl of 4-)-3-(O-MOM)-2-pyridine carboxamide (252mg, 80%).
1H?NMR(300MHz,CD
3OD)δ8.27-8.17(m,1H),7.75-7.64(m,1H),7.51-7.39(m,2H),7.33-7.24(m,1H),7.07(dt,J=21.4,8.8Hz,2H),5.29(d,J=14.0Hz,2H),4.79(s,1H),4.35(s,1H),3.45(dd,J=5.8,1.2Hz,3H),3.10-2.99(m,1H),1.61(dd,J=15.2,7.6Hz,1H),1.44(ddd,J=20.7,15.0,7.2Hz,2H),1.25(dd,J=14.7,7.5Hz,1H),1.07(dd,J=14.8,7.5Hz,1H),0.95(t,J=7.3Hz,1H),0.69(t,J=7.4Hz,2H).
Step (4): the preparation of N-normal-butyl-N-(the fluoro-benzyl of 4-)-3-hydroxyl-2-pyridine carboxamide (LZ-62)
N-normal-butyl-N-(the fluoro-benzyl of 4-)-3-(O-MOM)-2-pyridine carboxamide (250mg, 0.722mmol) be dissolved in 10ml methyl alcohol, ice bath to 0 ℃ stirring, Acetyl Chloride 98Min. (670 μ l, 19.8mmol) dropwise add in above-mentioned solution, continue ice bath and stir 10 minutes, be warming up to room temperature reaction 12 hours.Be spin-dried for solvent, column chromatography (petrol ether/ethyl acetate=10/1) obtains colourless oil liquid compound L Z-62 (170mg, 78%).
1H?NMR(300MHz,CD
3OD)δ8.16(s,1H),7.86(d,J=23.1Hz,2H),7.53(dd,J=8.6,5.3Hz,2H),7.19(t,J=8.7Hz,2H),4.19(s,2H),3.08-2.98(m,2H),1.75-1.62(m,2H),1.44(dt,J=14.9,7.3Hz,2H),0.98(t,J=7.3Hz,3H).ESI-MS:m/z?303.2[M+H]
+.
Embodiment 20
N-isobutyl--N-(the fluoro-benzyl of 4-)-3-hydroxyl-2-pyridine carboxamide (LZ-63)
Step (1): the preparation of N-isobutyl--3-hydroxyl-2-pyridine carboxamide
Synthetic method, with compound LZ-26, just changes raw material NSC 158269 into isobutylamine.Obtain faint yellow oily body compound N-isobutyl--3-hydroxyl-2-pyridine carboxamide (223mg, 80%).
Step (2): the preparation of N-isobutyl--3-(O-MOM)-2-pyridine carboxamide
Synthetic method, with compound LZ-60, just changes raw material N-(the fluoro-benzyl of 4-)-3-hydroxyl-2-pyridine carboxamide into N-isobutyl--3-hydroxyl-2-pyridine carboxamide.Obtain faint yellow oily body compound N-isobutyl--3-(O-MOM)-2-pyridine carboxamide (218mg, 72%).
1H?NMR(300MHz,CD
3OD)δ8.22(d,J=4.6Hz,1H),7.72(d,J=8.6Hz,1H),7.46(dd,J=8.6,4.6Hz,1H),5.30(s,2H),3.48(d,J=0.5Hz,3H),3.22(d,J=6.9Hz,2H),2.00-1.85(m,1H),0.99(d,J=6.7Hz,6H).
Step (3): the preparation of N-isobutyl--N-(the fluoro-benzyl of 4-)-3-(O-MOM)-2-pyridine carboxamide
Synthetic method, with compound LZ-61, just changes raw material N-normal-butyl-2-pyridine carboxamide into N-isobutyl--3-(O-MOM)-2-pyridine carboxamide.Obtain colorless oil compound N-isobutyl--N-(the fluoro-benzyl of 4-)-3-(O-MOM)-2-pyridine carboxamide (220mg, 81%).
1H?NMR(300MHz,CD
3OD)δ8.22(t,J=4.3Hz,2H),7.68(t,J=8.1Hz,2H),7.51-7.37(m,4H),7.30-7.22(m,2H),7.11(t,J=8.8Hz,2H),7.03(t,J=8.7Hz,2H),5.29(s,2H),5.27(s,2H),4.81(s,2H),4.37(s,2H),3.46(s,3H),3.43(s,3H),2.90(d,J=7.5Hz,2H),2.20-2.08(m,1H),1.94-1.83(m,1H),1.00(d,J=6.7Hz,6H),0.73(d,J=6.7Hz,6H).
Step (4): the preparation of N-isobutyl--N-(the fluoro-benzyl of 4-)-3-hydroxyl-2-pyridine carboxamide (LZ-63)
Synthetic method is with compound LZ-62, just changing raw material N-normal-butyl-N-(the fluoro-benzyl of 4-)-3-(O-MOM)-2-pyridine carboxamide into N-isobutyl--N-(the fluoro-benzyl of 4-)-3-(O-MOM)-2-pyridine carboxamide obtains colourless oil liquid compound L Z-63 (168mg, 77%).
1H?NMR(300MHz,CDCl
3)δ11.90(s,2H),8.12(d,J=27.1Hz,2H),7.82(d,J=29.2Hz,2H),7.48-7.29(m,3H),7.23(s,2H),7.00(t,J=7.7Hz,4H),5.08(s,2H),4.75(s,2H),3.57(s,2H),3.28(d,J=7.1Hz,2H),2.09(t,J=12.4Hz,1H),1.93(t,J=10.1Hz,1H),0.94(d,J=6.5Hz,6H),0.81(d,J=6.3Hz,5H).ESI-MS:m/z?303.2[M+H]
+.
Embodiment 21
N-(3-methyl-butyl)-N-(the fluoro-benzyl of 4-)-3-hydroxyl-2-pyridine carboxamide (LZ-64)
The preparation of step (1): N-(3-methyl-butyl)-3-hydroxyl-2-pyridine carboxamide
Synthetic method, with compound LZ-26, just changes raw material NSC 158269 into isobutylcarbylamine.Obtain faint yellow oily body compound N-(3-methyl-butyl)-3-hydroxyl-2-pyridine carboxamide (237mg, 82%).
The preparation of step (2): N-(3-methyl-butyl)-3-(O-MOM)-2-pyridine carboxamide
Synthetic method, with compound LZ-60, just changes raw material N-(the fluoro-benzyl of 4-)-3-hydroxyl-2-pyridine carboxamide into N-(3-methyl-butyl)-3-hydroxyl-2-pyridine carboxamide.Obtain faint yellow oily body compound N-(3-methyl-butyl)-3-(O-MOM)-2-pyridine carboxamide (220mg, 77%).
1H?NMR(300MHz,CD
3OD)δ8.22(d,J=4.6Hz,1H),7.72(d,J=8.6Hz,1H),7.46(dd,J=8.6,4.6Hz,1H),5.30(s,2H),3.48(d,J=0.5Hz,3H),3.22(d,J=6.9Hz,2H),2.00-1.85(m,1H),0.99(d,J=6.7Hz,6H).
The preparation of step (3): N-(3-methyl-butyl)-N-(the fluoro-benzyl of 4-)-3-(O-MOM)-2-pyridine carboxamide
Synthetic method, with compound LZ-61, just changes raw material N-normal-butyl-2-picolinamide into N-(3-methyl-butyl)-3-(O-MOM)-2-pyridine carboxamide.Obtain colorless oil compound N-(3-methyl-butyl)-N-(the fluoro-benzyl of 4-)-3-(O-MOM)-2-pyridine carboxamide (220mg, 81%).
1H?NMR(300MHz,CD
3OD)δ8.22(t,J=4.9Hz,1H),7.71(d,J=8.6Hz,1H),7.48(t,J=2.9Hz,2H),7.29(dd,J=8.3,5.3Hz,1H),7.11(t,J=8.8Hz,2H),5.32(s,2H),4.79(s,2H),3.45(s,3H),3.11-3.02(m,2H),1.51(dd,J=14.9,7.3Hz,1H),1.39(dd,J=15.2,7.2Hz,2H),0.62(d,J=6.5Hz,6H).
The preparation of step (4): N-(3-methyl-butyl)-N-(the fluoro-benzyl of 4-)-3-hydroxyl-2-pyridine carboxamide (LZ-64)
Synthetic method is with compound LZ-62, just changing raw material N-normal-butyl-N-(the fluoro-benzyl of 4-)-3-(O-MOM)-2-pyridine carboxamide into N-(3-methyl-butyl)-N-(the fluoro-benzyl of 4-)-3-(O-MOM)-2-pyridine carboxamide obtains colourless oil liquid compound L Z-64 (168mg, 77%).1H?NMR(300MHz,CD3OD)δ8.16(s,1H),7.87-7.84(m,2H),7.53(dd,J=8.4,5.2Hz,2H),7.19(t,J=8.5Hz,2H),4.19(s,2H),3.10-3.02(m,2H),1.74-1.64(m,1H),1.59(dd,J=7.2,1.2Hz,2H),0.96(d,J=6.3Hz,6H).ESI-MS:m/z?317.2[M+H]+.
Embodiment 22
N-(2-methacrylic)-N-(the fluoro-benzyl of 4-)-3-hydroxyl-2-pyridine carboxamide (LZ-65)
The preparation of step (1): N-(2-methacrylic)-3-hydroxyl-2-pyridine carboxamide
Synthetic method, with compound LZ-26, just changes raw material NSC 158269 into 2-methacrylic amine.Obtain faint yellow oily body compound N-(2-methacrylic)-3-hydroxyl-2-pyridine carboxamide (215mg, 78%).
The preparation of step (2): N-(2-methacrylic)-3-(O-MOM)-2-pyridine carboxamide
Synthetic method, with compound LZ-60, just changes raw material N-(the fluoro-benzyl of 4-)-3-hydroxyl-2-pyridine carboxamide into N-(2-methacrylic)-3-hydroxyl-2-pyridine carboxamide.Obtain faint yellow oily body compound N-(2-methacrylic)-3-(O-MOM)-2-pyridine carboxamide (141mg, 54%).
1H?NMR(300MHz,CD
3OD)δ8.23(d,J=4.6Hz,1H),7.72(d,J=8.6Hz,1H),7.47(dd,J=8.6,4.6Hz,1H),5.31(s,2H),4.98(s,2H),3.96(s,2H),3.48(s,3H),1.81(s,3H).
The preparation of step (3): N-(2-methacrylic)-(the fluoro-benzyl of 4-)-3-(O-MOM)-2-pyridine carboxamide
Synthetic method, with compound LZ-61, just changes raw material N-normal-butyl-2-pyridine carboxamide into N-(3-2-methacrylic)-3-(O-MOM)-2-pyridine carboxamide.Obtain colorless oil compound N-(2-methacrylic)-(the fluoro-benzyl of 4-)-3-(O-MOM)-2-picolinamide (165mg, 80%).
The preparation of step (4): N-(2-methacrylic)-N-(the fluoro-benzyl of 4-)-3-hydroxyl-2-pyridine carboxamide (LZ-65)
Synthetic method is with compound LZ-62, just changing raw material N-(2-methacrylic)-N-(the fluoro-benzyl of 4-)-3-(O-MOM)-2-pyridine carboxamide into N-(2-methacrylic)-(the fluoro-benzyl of 4-)-3-(O-MOM)-2-pyridine carboxamide obtains colourless oil liquid compound L Z-65 (126mg, 88%).
1H?NMR(300MHz,CDCl
3)δ12.10(s,2H),8.06(d,J=7.9Hz,2H),7.44(d,J=8.2Hz,2H),7.31(dd,J=8.4,4.5Hz,4H),7.01(t,J=8.9Hz,4H),5.29(s,2H),4.94(d,J=17.3Hz,2H),4.82(d,J=9.2Hz,2H),4.70(d,J=8.9Hz,4H),4.02(s,2H),1.76(s,3H),1.70(s,3H).
The experimental example of biologic activity
Biological reagent, chemical reagent and enzyme
All testing samples are dissolved in DMSO, and storage liquid concentration is 10mM, and condition of storage is :-20 ℃.Further dilution still be take DMSO as solvent.Be used for the expression system of purifying intergrase by America NI H, the Dr.Robert Craigie of Laboratory of Molecular Biology provides.Oligonucleotide for the test of intergrase catalytic activity is synthetic on the microsequencing core facility of USC Norris Cancer Center.γ [
32p]-ATP is purchased from Perkin Elmer (Waltham, MA).
Prepare oligonucleotide substrate
A 21 ' aggressiveness uplink is used in the test of intergrase catalytic activity:
(5 '-GTGTGGAAAATCTCTA GCAGT-3 ') and a descending chain of 21 ' aggressiveness:
(5’-ACTGCTAGAGATTTTCCACAC-3’)。By uplink 5 ' end by T4 polymerized nucleoside enzyme with [
32p]-ATP mark.By mark with unlabelled mixture of ribonucleotides in 95 ℃ hatching 15 minutes, make enzyme deactivation, then add the excessive descending chain of 1.5mol.Slowly cool to annealing at room temperature.Upper centrifugal in whizzer (Spin-25mini-column, USA Scientific, Ocala, FL), remove unconjugated dissociant.
Compound suppresses active and IN-LEDGF/p75 is suppressed to active test result intergrase dimerization
LEDGF/p75-IN AlphaScreen assay is according to Perkin-Elmer, and method is carried out described in Benelux company operational manual.Stock solution is diluted on 384 orifice plates, and every hole final volume is 25 μ l.Buffered soln is by 25mM Tris-HCl (pH 7.4), 150mM NaCl, 1mM MgCl
2, 0.01% (v/v) Tween-20 and 0.1% (w/v) bovin serum albumin are formulated.Testing sample is made into the gradient concentration not waiting to 100 μ M from 0.1 μ M, the testing sample of different concns adds in hand-hole and His
6the intergrase of-mark (final concentration 300nM) was in 4 ℃ of hatchings 30 minutes.The Flag-LEDGF/p75 that adds subsequently 100nM, continues hatching one hour in 4 ℃.Then add the acceptor magnetic bead of 5 μ l Ni inner complex parcels and the donor magnetic bead of the anti-Flag of 5 μ l, final concentration is 20 μ g/ml.The mixture of albumen and magnetic bead, in 30 ℃ of hatchings 1 hour, makes the two abundant combination.Avoid as much as possible light direct irradiation, observe and detect the utilizing emitted light of acceptor magnetic bead with EnVision plate reader (Perkin-Elmer), application EnVision management software carries out data analysis.
Intergrase dimerization active testing adopts AlphaScreen
method.Intergrase extract is dissolved in buffered soln (150mM NaCl, 25mM Tris-HCl pH7.3,1mM MgCl
2, 0.1% Tween 20,0.1%BSA).Stock solution is diluted on 384 orifice plates, and every hole final volume is 25 μ l.The saltant type intergrase of the wild-type intergrase of GST-mark and the 6xHis-mark of same concentrations spends the night in 4 ℃ of hatchings.Morning next day, the donor magnetic bead and the Ni that add gsh to wrap up
2+the acceptor magnetic bead (purchased from Perkin Elmer company) of parcel, final concentration is 1mg/mL.Culture plate was in 30 ℃ of hatchings 1 hour.With EnVision
tMplate reader (purchased from Perkin-Elmer company) scans.
Table 1. compound is active and active to the inhibition of IN-LEDGF/p75 combination to the inhibition of intergrase dimerization
The activity data demonstration of listing in table 1, the compound of general structure I representative of the present invention is that a class has the compound that suppresses active, the inhibition IC of representative compounds LZ-33 to IN-LEDGF/p75 and IN dimerization to intergrase dimerization and IN-LEDGF/p75
50be respectively 1.7uM, 4uM.Due to it, that the multiple site of intergrase and many processes are all had to good inhibition is active, and the mechanism of action is different from existing intergrase and suppresses medicine, can avoid the generation of cross resistance.This compounds novelty simple in structure, is worth carrying out further structure activity relationship exploration, has the potential quality of further investigation, is expected to develop into a class and can avoids the new texture integrase inhibitor with existing HIV-1 intergrase medicine crossing drug resistant.
In sum, the HIV-1 integrase inhibitor that the compounds of this invention is new texture, is expected to develop into the medicine of the disease (as acquired immune deficiency syndrome (AIDS), hepatitis B, the third liver) that comprises that the retroviral of HIV-1 intergrase is integrase mediated.
Claims (10)
1. pyridine-2-amides or its pharmacy acceptable salt shown in the following general formula I of a class,
Wherein,
L is: (1) key; (2) C
1-C
6alkylidene group; (3) C
2-C
6unsaturated alkylene; (4) (C
0-C
6alkylidene group)-(C
3-C
6cycloalkylidene)-(C
0-C
6alkylidene group); Or (5) (C
0-C
6alkylidene group)-M-(C
0-C
6alkylidene group), wherein, M is N (R
a), OC (=O) or C (=O) O; Wherein, the alkylidene group in the unsaturated alkylene in (3) and (2), (4), (5) not necessarily by 1-3 separately independently substituting group replace, described substituting group is selected among following groups: C
1-C
6alkyl, C
1-C
6alkoxy C
1-C
6alkyl, C
3-C
8cycloalkyl, halogen, sulfydryl, hydroxyl ,-CF
3,-CN ,-NO
2,-NR
ar
b,-NR
acOR
b,-NR
acOOR
b,-NR
asO
2r
b,-COOR
b,-COR
b,-CONR
ar
b,-SO
2r
b,-SO
2nR
ar
b,-OR
awith-OCOR
b;
R
1, R
2be hydrogen independently of one another; Hydroxyl; Or do not replace or by 1-3 the following groups that independently substituting group replaces separately: C
1-C
8alkyl, C
1-C
8alkoxy C
1-C
8alkyl, C
1-C
8alkoxyl group, C
2-C
8unsaturated alkyl, C
3-C
8cycloalkyl, C
6-C
12aryl, C
4-C
10heteroaryl or C
3-C
10heterocyclic radical; Described heteroaryl or heterocyclic radical comprise that 1-3 is selected from the heteroatoms in N, O and S; Described substituting group is selected from following groups: C
1-C
6alkyl, C
1-C
6alkoxyl group, C
1-C
6alkoxy C
1-C
6alkyl, halogen, amino, nitro, sulfydryl, hydroxyl ,-CN and-CF
3;
R
3for hydrogen, halogen, hydroxyl, sulfydryl ,-CF
3,-CN ,-NO
2, or do not replace or independently of one another by the following groups of 1-3 substituting group replacement: C
1-C
8alkyl, C
1-C
8alkoxyl group, C
2-C
8thiazolinyl, C
2-C
8alkynyl, C
3-C
8cycloalkyl, C
3-C
8cycloalkyloxy or amino; Described substituting group is selected among following groups: C
1-C
6alkyl, C
1-C
6alkoxyl group, C
1-C
6alkoxy C
1-C
6alkyl, halogen, sulfydryl, hydroxyl ,-CF
3,-CN ,-NO
2, ternary ,-NR amino to six-ring
ar
b,-NR
acOR
b,-NR
acOOR
b,-NR
asO
2r
b,-COOR
b,-COR
b,-CONR
ar
b,-SO
2r
b,-SO
2nR
ar
b,-OR
awith-OCOR
b; And R
3position be 3-position, 4-position, 5-position or the 6-position on pyridine ring;
Wherein, R
aand R
bbe hydrogen or C independently of one another
1-C
6alkyl;
Described halogen is fluorine, chlorine, bromine or iodine.
2. pyridine-2-amides according to claim 1 or its pharmacy acceptable salt, wherein,
L is C
1-C
4alkylidene group or C
2-C
4unsaturated alkylene;
R
1for hydrogen, C
1-C
8the alkyl of straight or branched or C
2-C
8the unsaturated alkyl of straight or branched;
R
2for hydrogen, C
1-C
8the alkyl of straight or branched, C
2-C
8the unsaturated alkyl of straight or branched, C
3-C
8cycloalkyl, do not replace or by 1-3 the C that independently substituting group replaces separately
6-C
12aryl, do not replace or by 1-3 the C that independently substituting group replaces separately
4-C
10heteroaryl or do not replace or by 1-3 the C that independently substituting group replaces separately
3-C
10heterocyclic radical, described heteroaryl or heterocyclic radical comprise 1-3 and are selected from the heteroatoms in N, O and S; Wherein, described substituting group is halogen, amino, nitro, hydroxyl, cyano group ,-CF
3, C
1-C
6alkoxyl group or C
1-C
6alkyl;
R
3for hydrogen, halogen, hydroxyl, sulfydryl ,-CF
3,-CN ,-NO
2, C
1-C
6alkyl or do not replace or by C
1-C
4the C that alkoxyl group replaces
1-C
6alkoxyl group.
3. pyridine-2-amides according to claim 2 or its pharmacy acceptable salt, wherein,
L is C
1-C
2alkylidene group;
R
1for hydrogen, C
1-C
6the alkyl of straight or branched or C
2-C
6the thiazolinyl of straight or branched;
R
2for hydrogen, the alkyl of C1-C6 straight or branched, C
3-C
6cycloalkyl, do not replace or by 1-3 the C that independently substituting group replaces separately
6-C
10aryl, do not replace or by 1-3 the C that independently substituting group replaces separately
4-C
9heteroaryl or do not replace or by 1-3 the C that independently substituting group replaces separately
3-C
8heterocyclic radical, described heteroaryl or heterocyclic radical comprise 1-3 and are selected from the heteroatoms in N, O and S; Wherein, described substituting group is halogen, amino, nitro, hydroxyl, cyano group ,-CF
3, C
1-C
4alkoxyl group or C
1-C
4alkyl;
R
3for hydrogen, halogen, hydroxyl, sulfydryl ,-CF
3,-CN ,-NO
2, C
1-C
4alkyl or do not replace or by C
1-C
2the C that alkoxyl group replaces
1-C
4alkoxyl group.
4. pyridine-2-amides according to claim 3 or its pharmacy acceptable salt, wherein,
L is methylene radical;
R
1for hydrogen, normal-butyl, isobutyl-, isopentyl or 2-methacrylic;
R
2for the alkyl of hydrogen, C1-C4 straight or branched, cyclohexyl, do not replace or by 1-3 phenyl, naphthyl, furyl, thienyl or the indyl that independently substituting group replaces separately; Wherein, described substituting group is halogen, amino, nitro, hydroxyl, cyano group ,-CF
3or methoxyl group;
R
3for hydrogen, halogen, hydroxyl, methoxyl group or methoxymethoxy.
5. pyridine-2-amides according to claim 1 or its pharmacy acceptable salt, wherein, described pyridine compounds and their is:
6. a method of preparing pyridine-2-amides claimed in claim 1, described method is undertaken by reaction path as follows:
Work as R
1during for H,
(a) compound 7-1 and amine
carry out condensation, obtain compound of Formula I;
Work as R
1while being not H,
(b) compound 7-1 and amine R
1nH
2carry out condensation, obtain compound 7-2;
(c) compound 7-2 and halohydrocarbon
there is nucleophilic reaction and obtain general formula compound I;
Wherein, X is fluorine, chlorine, bromine or iodine;
R
1, R
2, R
3identical with the definition in claim 1 with the definition of L.
In claim 1-5 the pyridine-2-amides described in any one or its pharmacy acceptable salt as the purposes of HIV-1 integrase inhibitor.
8. the purposes in the medicine that in claim 1-5, the pyridine-2-amides described in any one or its pharmacy acceptable salt are treated the integrase mediated disease by retroviral as integrase inhibitor in preparation.
9. purposes according to claim 8, wherein, described retroviral intergrase is HIV-1 intergrase.
10. a pharmaceutical composition, it comprises the pyridine-2-amides described in any one or its pharmacy acceptable salt and pharmaceutically acceptable carrier in one or more claims 1-5 that treats significant quantity.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
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| US20210114984A1 (en) * | 2018-05-25 | 2021-04-22 | Syngenta Participations Ag | Microbiocidal picolinamide derivatives |
| US11629129B2 (en) * | 2018-05-25 | 2023-04-18 | Syngenta Participations Ag | Microbiocidal picolinamide derivatives |
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