CN115385819A - 一种迷迭香酸生物电子等排体及其制备方法和应用 - Google Patents
一种迷迭香酸生物电子等排体及其制备方法和应用 Download PDFInfo
- Publication number
- CN115385819A CN115385819A CN202211159907.3A CN202211159907A CN115385819A CN 115385819 A CN115385819 A CN 115385819A CN 202211159907 A CN202211159907 A CN 202211159907A CN 115385819 A CN115385819 A CN 115385819A
- Authority
- CN
- China
- Prior art keywords
- reaction
- arh
- rosmarinic acid
- room temperature
- distilled water
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
- DOUMFZQKYFQNTF-WUTVXBCWSA-N (R)-rosmarinic acid Chemical compound C([C@H](C(=O)O)OC(=O)\C=C\C=1C=C(O)C(O)=CC=1)C1=CC=C(O)C(O)=C1 DOUMFZQKYFQNTF-WUTVXBCWSA-N 0.000 title claims abstract description 107
- ZZAFFYPNLYCDEP-HNNXBMFYSA-N Rosmarinsaeure Natural products OC(=O)[C@H](Cc1cccc(O)c1O)OC(=O)C=Cc2ccc(O)c(O)c2 ZZAFFYPNLYCDEP-HNNXBMFYSA-N 0.000 title claims abstract description 53
- DOUMFZQKYFQNTF-MRXNPFEDSA-N rosemarinic acid Natural products C([C@H](C(=O)O)OC(=O)C=CC=1C=C(O)C(O)=CC=1)C1=CC=C(O)C(O)=C1 DOUMFZQKYFQNTF-MRXNPFEDSA-N 0.000 title claims abstract description 53
- TVHVQJFBWRLYOD-UHFFFAOYSA-N rosmarinic acid Natural products OC(=O)C(Cc1ccc(O)c(O)c1)OC(=Cc2ccc(O)c(O)c2)C=O TVHVQJFBWRLYOD-UHFFFAOYSA-N 0.000 title claims abstract description 53
- 238000002360 preparation method Methods 0.000 title claims abstract description 23
- 108090000790 Enzymes Proteins 0.000 claims abstract description 12
- 102000004190 Enzymes Human genes 0.000 claims abstract description 12
- 238000006243 chemical reaction Methods 0.000 claims description 115
- 239000007787 solid Substances 0.000 claims description 91
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 claims description 75
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical class O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 67
- 150000001875 compounds Chemical class 0.000 claims description 47
- YMWUJEATGCHHMB-UHFFFAOYSA-N Dichloromethane Chemical compound ClCCl YMWUJEATGCHHMB-UHFFFAOYSA-N 0.000 claims description 45
- 239000012153 distilled water Substances 0.000 claims description 38
- 239000002904 solvent Substances 0.000 claims description 33
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 claims description 32
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 claims description 24
- 238000001914 filtration Methods 0.000 claims description 23
- 238000005406 washing Methods 0.000 claims description 22
- 238000005303 weighing Methods 0.000 claims description 22
- 238000001514 detection method Methods 0.000 claims description 21
- 230000000694 effects Effects 0.000 claims description 20
- 239000011734 sodium Substances 0.000 claims description 19
- 239000000243 solution Substances 0.000 claims description 17
- WMFOQBRAJBCJND-UHFFFAOYSA-M Lithium hydroxide Chemical compound [Li+].[OH-] WMFOQBRAJBCJND-UHFFFAOYSA-M 0.000 claims description 16
- OFOBLEOULBTSOW-UHFFFAOYSA-N Malonic acid Chemical compound OC(=O)CC(O)=O OFOBLEOULBTSOW-UHFFFAOYSA-N 0.000 claims description 16
- NQRYJNQNLNOLGT-UHFFFAOYSA-N Piperidine Chemical compound C1CCNCC1 NQRYJNQNLNOLGT-UHFFFAOYSA-N 0.000 claims description 16
- JUJWROOIHBZHMG-UHFFFAOYSA-N Pyridine Chemical compound C1=CC=NC=C1 JUJWROOIHBZHMG-UHFFFAOYSA-N 0.000 claims description 16
- 239000012065 filter cake Substances 0.000 claims description 15
- 230000002401 inhibitory effect Effects 0.000 claims description 15
- 238000003756 stirring Methods 0.000 claims description 15
- 239000003814 drug Substances 0.000 claims description 14
- 238000010438 heat treatment Methods 0.000 claims description 13
- 238000010992 reflux Methods 0.000 claims description 13
- 239000003826 tablet Substances 0.000 claims description 12
- VIMMECPCYZXUCI-MIMFYIINSA-N (4s,6r)-6-[(1e)-4,4-bis(4-fluorophenyl)-3-(1-methyltetrazol-5-yl)buta-1,3-dienyl]-4-hydroxyoxan-2-one Chemical compound CN1N=NN=C1C(\C=C\[C@@H]1OC(=O)C[C@@H](O)C1)=C(C=1C=CC(F)=CC=1)C1=CC=C(F)C=C1 VIMMECPCYZXUCI-MIMFYIINSA-N 0.000 claims description 11
- 241000187479 Mycobacterium tuberculosis Species 0.000 claims description 11
- 239000012043 crude product Substances 0.000 claims description 9
- 238000004090 dissolution Methods 0.000 claims description 9
- LLMLNAVBOAMOEE-UHFFFAOYSA-N 2,3-dichlorobenzaldehyde Chemical compound ClC1=CC=CC(C=O)=C1Cl LLMLNAVBOAMOEE-UHFFFAOYSA-N 0.000 claims description 8
- JGFZNNIVVJXRND-UHFFFAOYSA-N N,N-Diisopropylethylamine (DIPEA) Chemical compound CCN(C(C)C)C(C)C JGFZNNIVVJXRND-UHFFFAOYSA-N 0.000 claims description 8
- KAFHLONDOVSENM-HNNXBMFYSA-N O-Benzyl-L-tyrosine Chemical compound C1=CC(C[C@H](N)C(O)=O)=CC=C1OCC1=CC=CC=C1 KAFHLONDOVSENM-HNNXBMFYSA-N 0.000 claims description 8
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical class [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 claims description 8
- 210000002421 cell wall Anatomy 0.000 claims description 8
- 239000005457 ice water Substances 0.000 claims description 8
- 238000000034 method Methods 0.000 claims description 8
- 239000003208 petroleum Substances 0.000 claims description 8
- UMJSCPRVCHMLSP-UHFFFAOYSA-N pyridine Natural products COC1=CC=CN=C1 UMJSCPRVCHMLSP-UHFFFAOYSA-N 0.000 claims description 8
- 238000010898 silica gel chromatography Methods 0.000 claims description 8
- 239000000126 substance Substances 0.000 claims description 8
- 239000006185 dispersion Substances 0.000 claims description 7
- 238000001035 drying Methods 0.000 claims description 7
- SRCZQMGIVIYBBJ-UHFFFAOYSA-N ethoxyethane;ethyl acetate Chemical compound CCOCC.CCOC(C)=O SRCZQMGIVIYBBJ-UHFFFAOYSA-N 0.000 claims description 7
- 238000002347 injection Methods 0.000 claims description 7
- 239000007924 injection Substances 0.000 claims description 7
- 239000000843 powder Substances 0.000 claims description 7
- 229920002472 Starch Polymers 0.000 claims description 6
- 239000002775 capsule Substances 0.000 claims description 6
- 238000001816 cooling Methods 0.000 claims description 6
- 239000000839 emulsion Substances 0.000 claims description 6
- HQKMJHAJHXVSDF-UHFFFAOYSA-L magnesium stearate Chemical compound [Mg+2].CCCCCCCCCCCCCCCCCC([O-])=O.CCCCCCCCCCCCCCCCCC([O-])=O HQKMJHAJHXVSDF-UHFFFAOYSA-L 0.000 claims description 6
- 239000006187 pill Substances 0.000 claims description 6
- 239000008107 starch Substances 0.000 claims description 6
- 235000019698 starch Nutrition 0.000 claims description 6
- 239000000829 suppository Substances 0.000 claims description 6
- 229940079593 drug Drugs 0.000 claims description 5
- 238000000605 extraction Methods 0.000 claims description 5
- 230000001355 anti-mycobacterial effect Effects 0.000 claims description 4
- 230000015572 biosynthetic process Effects 0.000 claims description 4
- 238000005507 spraying Methods 0.000 claims description 4
- DGAQECJNVWCQMB-PUAWFVPOSA-M Ilexoside XXIX Chemical compound C[C@@H]1CC[C@@]2(CC[C@@]3(C(=CC[C@H]4[C@]3(CC[C@@H]5[C@@]4(CC[C@@H](C5(C)C)OS(=O)(=O)[O-])C)C)[C@@H]2[C@]1(C)O)C)C(=O)O[C@H]6[C@@H]([C@H]([C@@H]([C@H](O6)CO)O)O)O.[Na+] DGAQECJNVWCQMB-PUAWFVPOSA-M 0.000 claims description 3
- GUBGYTABKSRVRQ-QKKXKWKRSA-N Lactose Natural products OC[C@H]1O[C@@H](O[C@H]2[C@H](O)[C@@H](O)C(O)O[C@@H]2CO)[C@H](O)[C@@H](O)[C@H]1O GUBGYTABKSRVRQ-QKKXKWKRSA-N 0.000 claims description 3
- 229920000168 Microcrystalline cellulose Polymers 0.000 claims description 3
- 125000002057 carboxymethyl group Chemical group [H]OC(=O)C([H])([H])[*] 0.000 claims description 3
- 239000011248 coating agent Substances 0.000 claims description 3
- 238000000576 coating method Methods 0.000 claims description 3
- 239000001866 hydroxypropyl methyl cellulose Substances 0.000 claims description 3
- 235000010979 hydroxypropyl methyl cellulose Nutrition 0.000 claims description 3
- 229920003088 hydroxypropyl methyl cellulose Polymers 0.000 claims description 3
- 239000008101 lactose Substances 0.000 claims description 3
- 235000019359 magnesium stearate Nutrition 0.000 claims description 3
- 235000019813 microcrystalline cellulose Nutrition 0.000 claims description 3
- 239000008108 microcrystalline cellulose Substances 0.000 claims description 3
- 229940016286 microcrystalline cellulose Drugs 0.000 claims description 3
- 238000002156 mixing Methods 0.000 claims description 3
- 230000008569 process Effects 0.000 claims description 3
- 239000000047 product Substances 0.000 claims description 3
- 229910052708 sodium Inorganic materials 0.000 claims description 3
- 238000009775 high-speed stirring Methods 0.000 claims description 2
- 229960003943 hypromellose Drugs 0.000 claims description 2
- 238000005453 pelletization Methods 0.000 claims description 2
- 239000000376 reactant Substances 0.000 claims description 2
- 238000001291 vacuum drying Methods 0.000 claims 2
- 239000002671 adjuvant Substances 0.000 claims 1
- 238000000338 in vitro Methods 0.000 abstract description 11
- 241000186366 Mycobacterium bovis Species 0.000 abstract description 10
- 238000002474 experimental method Methods 0.000 abstract description 4
- 238000005160 1H NMR spectroscopy Methods 0.000 description 33
- 238000002330 electrospray ionisation mass spectrometry Methods 0.000 description 33
- 238000001644 13C nuclear magnetic resonance spectroscopy Methods 0.000 description 26
- 201000008827 tuberculosis Diseases 0.000 description 14
- 230000002365 anti-tubercular Effects 0.000 description 11
- 239000000523 sample Substances 0.000 description 11
- 125000001931 aliphatic group Chemical group 0.000 description 10
- 229910052736 halogen Inorganic materials 0.000 description 8
- 210000002540 macrophage Anatomy 0.000 description 8
- -1 (4-bromo) -phenylalanyl Chemical group 0.000 description 6
- 125000002252 acyl group Chemical group 0.000 description 6
- 125000003118 aryl group Chemical group 0.000 description 6
- 150000002367 halogens Chemical class 0.000 description 6
- 239000000463 material Substances 0.000 description 6
- 230000000877 morphologic effect Effects 0.000 description 6
- 125000001997 phenyl group Chemical group [H]C1=C([H])C([H])=C(*)C([H])=C1[H] 0.000 description 6
- 239000004480 active ingredient Substances 0.000 description 5
- 238000013461 design Methods 0.000 description 5
- 239000000203 mixture Substances 0.000 description 5
- 238000005086 pumping Methods 0.000 description 5
- IAZDPXIOMUYVGZ-WFGJKAKNSA-N Dimethyl sulfoxide Chemical compound [2H]C([2H])([2H])S(=O)C([2H])([2H])[2H] IAZDPXIOMUYVGZ-WFGJKAKNSA-N 0.000 description 4
- ZQLQOXLUCGXKHS-SIAUPFDVSA-N UDP-alpha-D-galactofuranose Chemical compound O[C@@H]1[C@@H](O)[C@H]([C@H](O)CO)O[C@@H]1OP(O)(=O)OP(O)(=O)OC[C@@H]1[C@@H](O)[C@@H](O)[C@H](N2C(NC(=O)C=C2)=O)O1 ZQLQOXLUCGXKHS-SIAUPFDVSA-N 0.000 description 4
- HSCJRCZFDFQWRP-ABVWGUQPSA-N UDP-alpha-D-galactose Chemical compound O[C@@H]1[C@@H](O)[C@@H](O)[C@@H](CO)O[C@@H]1OP(O)(=O)OP(O)(=O)OC[C@@H]1[C@@H](O)[C@@H](O)[C@H](N2C(NC(=O)C=C2)=O)O1 HSCJRCZFDFQWRP-ABVWGUQPSA-N 0.000 description 4
- YLEIFZAVNWDOBM-ZTNXSLBXSA-N ac1l9hc7 Chemical compound C([C@H]12)C[C@@H](C([C@@H](O)CC3)(C)C)[C@@]43C[C@@]14CC[C@@]1(C)[C@@]2(C)C[C@@H]2O[C@]3(O)[C@H](O)C(C)(C)O[C@@H]3[C@@H](C)[C@H]12 YLEIFZAVNWDOBM-ZTNXSLBXSA-N 0.000 description 4
- 150000001338 aliphatic hydrocarbons Chemical class 0.000 description 4
- 125000000623 heterocyclic group Chemical group 0.000 description 4
- 229910052739 hydrogen Inorganic materials 0.000 description 4
- 239000001257 hydrogen Substances 0.000 description 4
- 208000015181 infectious disease Diseases 0.000 description 4
- 201000009671 multidrug-resistant tuberculosis Diseases 0.000 description 4
- 125000004433 nitrogen atom Chemical group N* 0.000 description 4
- 125000004430 oxygen atom Chemical group O* 0.000 description 4
- 239000011541 reaction mixture Substances 0.000 description 4
- 125000000472 sulfonyl group Chemical group *S(*)(=O)=O 0.000 description 4
- 125000004434 sulfur atom Chemical group 0.000 description 4
- 238000012360 testing method Methods 0.000 description 4
- 241000283690 Bos taurus Species 0.000 description 3
- 238000005481 NMR spectroscopy Methods 0.000 description 3
- 230000009471 action Effects 0.000 description 3
- 229940072185 drug for treatment of tuberculosis Drugs 0.000 description 3
- 238000011156 evaluation Methods 0.000 description 3
- 230000005764 inhibitory process Effects 0.000 description 3
- 238000012986 modification Methods 0.000 description 3
- 230000004048 modification Effects 0.000 description 3
- 239000002547 new drug Substances 0.000 description 3
- 238000012827 research and development Methods 0.000 description 3
- 150000003839 salts Chemical class 0.000 description 3
- 238000012216 screening Methods 0.000 description 3
- IQHSSYROJYPFDV-UHFFFAOYSA-N 2-bromo-1,3-dichloro-5-(trifluoromethyl)benzene Chemical group FC(F)(F)C1=CC(Cl)=C(Br)C(Cl)=C1 IQHSSYROJYPFDV-UHFFFAOYSA-N 0.000 description 2
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 description 2
- UFHFLCQGNIYNRP-UHFFFAOYSA-N Hydrogen Chemical compound [H][H] UFHFLCQGNIYNRP-UHFFFAOYSA-N 0.000 description 2
- HSCJRCZFDFQWRP-UHFFFAOYSA-N Uridindiphosphoglukose Natural products OC1C(O)C(O)C(CO)OC1OP(O)(=O)OP(O)(=O)OCC1C(O)C(O)C(N2C(NC(=O)C=C2)=O)O1 HSCJRCZFDFQWRP-UHFFFAOYSA-N 0.000 description 2
- 125000000217 alkyl group Chemical group 0.000 description 2
- 238000003556 assay Methods 0.000 description 2
- AVVWPBAENSWJCB-DGPNFKTASA-N beta-D-galactofuranose Chemical compound OC[C@@H](O)[C@@H]1O[C@@H](O)[C@H](O)[C@H]1O AVVWPBAENSWJCB-DGPNFKTASA-N 0.000 description 2
- 238000001460 carbon-13 nuclear magnetic resonance spectrum Methods 0.000 description 2
- 239000003153 chemical reaction reagent Substances 0.000 description 2
- 231100000433 cytotoxic Toxicity 0.000 description 2
- 238000011161 development Methods 0.000 description 2
- 239000002552 dosage form Substances 0.000 description 2
- 125000005843 halogen group Chemical group 0.000 description 2
- 125000005842 heteroatom Chemical group 0.000 description 2
- 150000002431 hydrogen Chemical class 0.000 description 2
- 239000003112 inhibitor Substances 0.000 description 2
- 238000001819 mass spectrum Methods 0.000 description 2
- 229930014626 natural product Natural products 0.000 description 2
- 125000000449 nitro group Chemical group [O-][N+](*)=O 0.000 description 2
- 239000000546 pharmaceutical excipient Substances 0.000 description 2
- 230000000144 pharmacologic effect Effects 0.000 description 2
- 230000035755 proliferation Effects 0.000 description 2
- 238000000425 proton nuclear magnetic resonance spectrum Methods 0.000 description 2
- 238000011160 research Methods 0.000 description 2
- 239000012453 solvate Substances 0.000 description 2
- 230000009897 systematic effect Effects 0.000 description 2
- LJIOTBMDLVHTBO-CUYJMHBOSA-N (2s)-2-amino-n-[(1r,2r)-1-cyano-2-[4-[4-(4-methylpiperazin-1-yl)sulfonylphenyl]phenyl]cyclopropyl]butanamide Chemical compound CC[C@H](N)C(=O)N[C@]1(C#N)C[C@@H]1C1=CC=C(C=2C=CC(=CC=2)S(=O)(=O)N2CCN(C)CC2)C=C1 LJIOTBMDLVHTBO-CUYJMHBOSA-N 0.000 description 1
- VUDZSIYXZUYWSC-DBRKOABJSA-N (4r)-1-[(2r,4r,5r)-3,3-difluoro-4-hydroxy-5-(hydroxymethyl)oxolan-2-yl]-4-hydroxy-1,3-diazinan-2-one Chemical compound FC1(F)[C@H](O)[C@@H](CO)O[C@H]1N1C(=O)N[C@H](O)CC1 VUDZSIYXZUYWSC-DBRKOABJSA-N 0.000 description 1
- IGVKWAAPMVVTFX-BUHFOSPRSA-N (e)-octadec-5-en-7,9-diynoic acid Chemical compound CCCCCCCCC#CC#C\C=C\CCCC(O)=O IGVKWAAPMVVTFX-BUHFOSPRSA-N 0.000 description 1
- SJHDFJKJPCOPNJ-UHFFFAOYSA-N 3-(2,3-dichlorophenyl)propanamide Chemical group NC(=O)CCC1=CC=CC(Cl)=C1Cl SJHDFJKJPCOPNJ-UHFFFAOYSA-N 0.000 description 1
- 241000193830 Bacillus <bacterium> Species 0.000 description 1
- 208000035473 Communicable disease Diseases 0.000 description 1
- AVVWPBAENSWJCB-RSVSWTKNSA-N D-galactofuranose Chemical compound OC[C@@H](O)[C@@H]1OC(O)[C@H](O)[C@H]1O AVVWPBAENSWJCB-RSVSWTKNSA-N 0.000 description 1
- 206010062016 Immunosuppression Diseases 0.000 description 1
- 101001073833 Klebsiella pneumoniae UDP-galactopyranose mutase Proteins 0.000 description 1
- 241000124008 Mammalia Species 0.000 description 1
- HSHXDCVZWHOWCS-UHFFFAOYSA-N N'-hexadecylthiophene-2-carbohydrazide Chemical compound CCCCCCCCCCCCCCCCNNC(=O)c1cccs1 HSHXDCVZWHOWCS-UHFFFAOYSA-N 0.000 description 1
- 229940125907 SJ995973 Drugs 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
- 239000000654 additive Substances 0.000 description 1
- 230000000259 anti-tumor effect Effects 0.000 description 1
- 230000002155 anti-virotic effect Effects 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000006696 biosynthetic metabolic pathway Effects 0.000 description 1
- 230000001472 cytotoxic effect Effects 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 239000003596 drug target Substances 0.000 description 1
- 208000015355 drug-resistant tuberculosis Diseases 0.000 description 1
- 230000009977 dual effect Effects 0.000 description 1
- 230000009982 effect on human Effects 0.000 description 1
- 230000005284 excitation Effects 0.000 description 1
- 208000036984 extensively drug-resistant tuberculosis Diseases 0.000 description 1
- 238000002875 fluorescence polarization Methods 0.000 description 1
- 239000007850 fluorescent dye Substances 0.000 description 1
- 238000009472 formulation Methods 0.000 description 1
- 229930182830 galactose Natural products 0.000 description 1
- 229960003082 galactose Drugs 0.000 description 1
- 125000002887 hydroxy group Chemical group [H]O* 0.000 description 1
- 230000001506 immunosuppresive effect Effects 0.000 description 1
- 238000001727 in vivo Methods 0.000 description 1
- 229940042040 innovative drug Drugs 0.000 description 1
- 150000002632 lipids Chemical class 0.000 description 1
- 239000004973 liquid crystal related substance Substances 0.000 description 1
- 231100000053 low toxicity Toxicity 0.000 description 1
- 239000011159 matrix material Substances 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- 230000007246 mechanism Effects 0.000 description 1
- IQKXGACPIPNLEL-NTISSMGPSA-N methyl (2s)-2-amino-3-(4-phenylmethoxyphenyl)propanoate;hydrochloride Chemical compound Cl.C1=CC(C[C@H](N)C(=O)OC)=CC=C1OCC1=CC=CC=C1 IQKXGACPIPNLEL-NTISSMGPSA-N 0.000 description 1
- 244000000010 microbial pathogen Species 0.000 description 1
- XZMHJYWMCRQSSI-UHFFFAOYSA-N n-[5-[2-(3-acetylanilino)-1,3-thiazol-4-yl]-4-methyl-1,3-thiazol-2-yl]benzamide Chemical compound CC(=O)C1=CC=CC(NC=2SC=C(N=2)C2=C(N=C(NC(=O)C=3C=CC=CC=3)S2)C)=C1 XZMHJYWMCRQSSI-UHFFFAOYSA-N 0.000 description 1
- 239000002674 ointment Substances 0.000 description 1
- 229910052760 oxygen Inorganic materials 0.000 description 1
- 244000052769 pathogen Species 0.000 description 1
- 230000001717 pathogenic effect Effects 0.000 description 1
- ISWSIDIOOBJBQZ-UHFFFAOYSA-N phenol group Chemical group C1(=CC=CC=C1)O ISWSIDIOOBJBQZ-UHFFFAOYSA-N 0.000 description 1
- 239000011505 plaster Substances 0.000 description 1
- 229920000642 polymer Polymers 0.000 description 1
- 230000002265 prevention Effects 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- 238000000746 purification Methods 0.000 description 1
- 239000002994 raw material Substances 0.000 description 1
- 238000007423 screening assay Methods 0.000 description 1
- 230000004083 survival effect Effects 0.000 description 1
- 238000003786 synthesis reaction Methods 0.000 description 1
- 231100000419 toxicity Toxicity 0.000 description 1
- 230000001988 toxicity Effects 0.000 description 1
- 229940126680 traditional chinese medicines Drugs 0.000 description 1
- 230000035899 viability Effects 0.000 description 1
Images
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
- C07C233/00—Carboxylic acid amides
- C07C233/01—Carboxylic acid amides having carbon atoms of carboxamide groups bound to hydrogen atoms or to acyclic carbon atoms
- C07C233/45—Carboxylic acid amides having carbon atoms of carboxamide groups bound to hydrogen atoms or to acyclic carbon atoms having the nitrogen atom of at least one of the carboxamide groups bound to a carbon atom of a hydrocarbon radical substituted by carboxyl groups
- C07C233/46—Carboxylic acid amides having carbon atoms of carboxamide groups bound to hydrogen atoms or to acyclic carbon atoms having the nitrogen atom of at least one of the carboxamide groups bound to a carbon atom of a hydrocarbon radical substituted by carboxyl groups with the substituted hydrocarbon radical bound to the nitrogen atom of the carboxamide group by an acyclic carbon atom
- C07C233/51—Carboxylic acid amides having carbon atoms of carboxamide groups bound to hydrogen atoms or to acyclic carbon atoms having the nitrogen atom of at least one of the carboxamide groups bound to a carbon atom of a hydrocarbon radical substituted by carboxyl groups with the substituted hydrocarbon radical bound to the nitrogen atom of the carboxamide group by an acyclic carbon atom having the carbon atom of the carboxamide group bound to an acyclic carbon atom of a carbon skeleton containing six-membered aromatic rings
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/20—Pills, tablets, discs, rods
- A61K9/28—Dragees; Coated pills or tablets, e.g. with film or compression coating
- A61K9/2806—Coating materials
- A61K9/282—Organic compounds, e.g. fats
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
- A61P31/04—Antibacterial agents
- A61P31/06—Antibacterial agents for tuberculosis
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
- C07C227/00—Preparation of compounds containing amino and carboxyl groups bound to the same carbon skeleton
- C07C227/14—Preparation of compounds containing amino and carboxyl groups bound to the same carbon skeleton from compounds containing already amino and carboxyl groups or derivatives thereof
- C07C227/18—Preparation of compounds containing amino and carboxyl groups bound to the same carbon skeleton from compounds containing already amino and carboxyl groups or derivatives thereof by reactions involving amino or carboxyl groups, e.g. hydrolysis of esters or amides, by formation of halides, salts or esters
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
- C07C231/00—Preparation of carboxylic acid amides
- C07C231/02—Preparation of carboxylic acid amides from carboxylic acids or from esters, anhydrides, or halides thereof by reaction with ammonia or amines
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
- C07C231/00—Preparation of carboxylic acid amides
- C07C231/12—Preparation of carboxylic acid amides by reactions not involving the formation of carboxamide groups
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
- C07C231/00—Preparation of carboxylic acid amides
- C07C231/22—Separation; Purification; Stabilisation; Use of additives
- C07C231/24—Separation; Purification
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
- C07C51/00—Preparation of carboxylic acids or their salts, halides or anhydrides
- C07C51/16—Preparation of carboxylic acids or their salts, halides or anhydrides by oxidation
Landscapes
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Health & Medical Sciences (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Veterinary Medicine (AREA)
- Animal Behavior & Ethology (AREA)
- Engineering & Computer Science (AREA)
- Public Health (AREA)
- Medicinal Chemistry (AREA)
- General Health & Medical Sciences (AREA)
- Pharmacology & Pharmacy (AREA)
- Life Sciences & Earth Sciences (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Communicable Diseases (AREA)
- General Chemical & Material Sciences (AREA)
- Pulmonology (AREA)
- Oncology (AREA)
- Oil, Petroleum & Natural Gas (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Epidemiology (AREA)
- Acyclic And Carbocyclic Compounds In Medicinal Compositions (AREA)
- Organic Low-Molecular-Weight Compounds And Preparation Thereof (AREA)
Abstract
本发明涉及一种迷迭香酸生物电子等排体及其制备方法和应用,通过生物电子等排策略,引入酰胺键代替迷迭香酸的酯键,设计并制备了一系列衍生物,结合酶水平和体外抗牛型结核分枝杆菌实验,表明本发明中涉及的化合物比母体化合物迷迭香酸表现出较佳的抗结核活性,可以作为药物活性物质开发抗结核杆菌药物,因此本发明为寻找具有良好开发前景的抗结核候选药物分子奠定基础。
Description
技术领域
本发明涉及药物化学领域,具体涉及一种迷迭香酸生物电子等排体及其制备方法和应用。
背景技术
结核病(Tuberculosis)是目前第二大单一感染源致死病因。结核病耐药结核病持续增多,据统计全球耐多药结核病(multidrug-resistant tuberculosis,MDR-TB)治疗成功率仅为59%,这也是直接导致目前传统治疗结核病的药物失去效力造成治愈率下降而且费用增高的原因。近年来,随着耐多药(MDR-TB)和泛耐药(extensively drug- resistanttuberculosis,XDR-TB)结核菌株的迅速增加,结核病与HIV并发感染等情况的出现,对本已严峻的结核病防控形势带来了新挑战。具有全新作用机制及高效低毒的抗结核新药的研发将是解决该传染病控制问题的最佳途径之一。
结核分枝杆菌的细胞壁在其生存和增殖的过程中起着至关重要的作用,近年来发现的有潜力的抗结核新药德拉马尼作用靶标也在细胞壁上,是通过抑制细胞壁霉菌酸的生物合成;2019年底发布的新药普托马尼PA-824具有抑制细胞壁脂质和蛋白质合成的双重功能。通过对细胞壁微观结构的深入研究来寻找抗结核药物的新靶标并筛选其对应的抑制剂,仍是抗结核新药研究的热点。
半乳呋喃糖(Galf)是包括结核分枝杆菌在内的许多致病微生物的重要组成部分,对病原体的生存能力至关重要。结核分枝杆菌细胞壁核心部分为mycolylarabinogalactan–peptidoglycan(mAPG)复合物,含有一种半乳糖聚合物(Galactan),大约由35个Galf组成。UDP—半乳糖变异酶(UDP-Galactopyranose Mutase,简称UGM),则是UDP-半乳呋喃糖(UDP-Galf)形成的关键酶之一,该酶能催化UDP-半乳吡喃糖(UDP-Galp)和UDP-半乳呋喃糖(UDP-Galf)之间的相互转化,抑制该酶的活性即有可能阻断该生物合成途径,进而抑制结核分枝杆菌的生长。目前已有相关研究证实通过抑制UGM的活性的确能有效阻止结核分枝杆菌的繁殖。更重要的是,由于UGM不存在哺乳动物体内,这意味着以该酶为靶标的特异性药物对治疗结核病将具有高度专一性且对人类不会有太大的毒性或副作用,因此,UGM可作为一个全新的抗结核理想药物靶标。
发明团队发现迷迭香酸显示出较好的抑制肺炎克雷伯氏菌UGM酶的活性,但是该化合物对结核分枝杆菌UGM酶表现出中等强度的抑制活性。迷迭香酸及其衍生物是多种中药中的有效活性成分,具有抑菌、抗肿瘤、抗病毒、免疫抑制等多种药理作用,一直以来都备受关注。据文献报道,有学者以迷迭香酸为母核设计了一系列衍生物,表现出较好的体外抗结核杆菌活性,显示出其潜在的临床应用价值。但由于迷迭香酸分子结构中含有酯键、酚羟基,致使其稳定性较差,在体内易水解从而导致其活性不佳,且迷迭香酸又具有广泛的药理活性而缺少选择性,目前尚未见迷迭香酸生物电子等排体在抑制UGM活性和抗牛型结核分枝杆菌的报道。
针对以上问题,本发明团队以天然产物迷迭香酸为母体,引入酯键生物电子等排体——酰胺键来重新构建该苗头化合物,对所获得的一系列衍生物进行酶水平和菌株水平的系统抗结核活性评价。力争从中找到具有良好开发前景的候选药物分子,为抗结核创新药物的研发奠定基础。
发明内容
本发明的目的是提供一种迷迭香酸生物电子等排体。
本发明的另一目的是提一种迷迭香酸生物电子等排体的其制备方法。
本发明的另一目的是提一种迷迭香酸生物电子等排体在制备抗牛型结核分枝杆菌药物中的应用。
本发明所述的迷迭香酸生物电子等排体,结构通式如下:
本发明所述的迷迭香酸生物电子等排体为下列化合物:
符合结构通式1的衍生物为:
符合结构通式2的衍生物为:
本发明所述的迷迭香酸生物电子等排体包括单键和双键。
本发明所述的迷迭香酸生物电子等排体的制备方法步骤为:
a.称取2.0-3.0g,10mmoL,1eq的O-苄基-L-酪氨酸于反应瓶中,向瓶内注入50-80mL甲醇;将反应瓶置于冰盐浴下冷却10-30min;待体系温度降低至-10℃后,向反应瓶内缓慢滴加4.4mL,60mmoL,6eq重蒸的SOCl2,控制滴速,使体系温度保持在 0℃以下;滴加完毕后,冰盐浴下反应5-15min,撤去冰盐浴后使反应瓶恢复至室温,再于60-100℃油浴下加热回流2-5h,TLC跟踪检测。反应结束后,减压回收溶剂,过滤,滤饼用二氯甲烷洗涤以获得所需化合物,将化合物真空抽干数小时后即得白色粉末状1;
b.称取5.0-6.0g,30mmoL,1eq的2,3-二氯苯甲醛于圆底烧瓶中,加入20-50mL 吡啶并于80-100℃油浴中搅拌溶解样品;溶解后向反应瓶中加入9.0-10.0g,90mmoL, 3eq的丙二酸并滴加880μL,9mmoL,0.3eq的哌啶于80-100℃油浴中加热回流2-5h 后,TLC检测反应结束;反应体系冷却至室温后,将其倾入200-500mL冰水溶液中,用5-15%的盐酸调节pH值至4,有大量的白色固体析出;过滤,白色固体用蒸馏水洗 2-3次;再用适量乙酸乙酯溶解白色固体,滤去不溶物;减压回收部分溶剂后置于冰箱中析出白色粉末状固体2;
c.称取0.4-0.5g,2mmoL,1eq的白色粉末状固体2置于反应瓶内,溶解于10-30 mL无水DMF中,再加入0.5-0.6g,2mmoL,1eq的HBTU和695μL,4mmoL,2eq的 DIEA,室温下反应20-50min;随后向反应物中加入0.5-0.6g,2mmoL,1eq的1,并于室温下反应10-15h;TLC检测反应结束后,加入50-100mL乙酸乙酯和20-50mL蒸馏水分散,萃取,依次以蒸馏水和饱和氯化钠溶液洗涤,无水MgSO4干燥,减压回收乙酸乙酯至干所得粗品经比例为3:1的石油醚:乙酸乙酯硅胶柱层析纯化,得白色固体3;
d.取白色固体3,加入5-10mL蒸馏水和30-40mg,1.5mmoL,1.5eq的LiOH,室温搅拌反应1-3h,TLC检测反应结束;用1M浓盐酸调pH至2-3;减压回收溶剂至干,所得残留物再经二氯甲烷洗涤后过滤,抽干滤饼得化合物4a,(N)-2-[(5,6-二氯基)- 苯丙烯酰胺基]-3-(4-苄氧基苯基)-丙酸。
优选的,
本发明所述的迷迭香酸生物电子等排体的制备方法步骤为:
a.称取2.71g,10mmoL,1eq的O-苄基-L-酪氨酸于反应瓶中,向瓶内注入60mL 甲醇;将反应瓶置于冰盐浴下冷却20min;待体系温度降低至-10℃后,向反应瓶内缓慢滴加4.4mL,60mmoL,6eq重蒸的SOCl2,控制滴速,使体系温度保持在0℃以下;滴加完毕后,冰盐浴下反应10min,撤去冰盐浴后使反应瓶恢复至室温,再于80℃油浴下加热回流3h,TLC跟踪检测。反应结束后,减压回收溶剂,过滤,滤饼用二氯甲烷洗涤以获得所需化合物,将化合物真空抽干数小时后即得白色粉末状1;
b.称取5.26g,30mmoL,1eq的2,3-二氯苯甲醛于圆底烧瓶中,加入30mL吡啶并于100℃油浴中搅拌溶解样品;溶解后向反应瓶中加入9.36g,90mmoL,3eq的丙二酸并滴加880μL,9mmoL,0.3eq的哌啶于100℃油浴中加热回流3h后,TLC检测反应结束;反应体系冷却至室温后,将其倾入300mL冰水溶液中,用10%的盐酸调节pH值至4,有大量的白色固体析出;过滤,白色固体用蒸馏水洗2-3次;再用适量乙酸乙酯溶解白色固体,滤去不溶物;减压回收部分溶剂后置于冰箱中析出白色粉末状固体2;
c.称取0.44g,2mmoL,1eq的白色粉末状固体2置于反应瓶内,溶解于20mL无水DMF中,再加入0.58g,2mmoL,1eq的HBTU和695μL,4mmoL,2eq的DIEA,室温下反应30min;随后向反应物中加入0.57g,2mmoL,1eq的1,并于室温下反应 14h;TLC检测反应结束后,加入80mL乙酸乙酯和40mL蒸馏水分散,萃取,依次以蒸馏水和饱和氯化钠溶液洗涤,无水MgSO4干燥,减压回收乙酸乙酯至干所得粗品经比例为3:1的石油醚:乙酸乙酯硅胶柱层析纯化,得白色固体3;
d.取白色固体3,加入5mL蒸馏水和36mg,1.5mmoL,1.5eq的LiOH,室温搅拌反应2h,TLC检测反应结束;用1M浓盐酸调pH至2-3;减压回收溶剂至干,所得残留物再经二氯甲烷洗涤后过滤,抽干滤饼得化合物4a,(N)-2-[(5,6-二氯基)-苯丙烯酰胺基]-3-(4-苄氧基苯基)-丙酸。
本发明所述制备方法可用于所述衍生物的制备。
本发明所述的迷迭香酸生物电子等排体衍生物加入药学上可接受的辅料制成药学上可接受的制剂;所述制剂为片剂、丸剂、胶囊剂、栓剂、乳剂和注射剂。
本发明所述片剂制备方法为:取任一衍生物的产物5g、羟丙甲纤维素6g、羧甲淀粉钠10g、微晶纤维素8g、乳糖115g、淀粉50g、硬脂酸镁1g;将主药与辅料充分混匀后投入高速搅拌机中,喷雾加水适量,整粒,水分控制在3~4%,然后压片,制成1000片,包薄膜衣。
本发明所述的迷迭香酸生物电子等排体在制备抑制结核分枝杆菌细胞壁生物合成关键酶——UGM活性及在制备抗牛型结核分枝杆菌药物中的应用。
本发明所述BCG菌株抑制浓度为5μg/mL-500μg/mL。
喷雾加水量为主药与辅料总量的1-5%或者适量。
本发明有益效果:
1.本发明以天然产物迷迭香酸为母体,首次将酯键生物电子等排体——酰胺键来重新构建迷迭香酸生物电子等排体一系列衍生物,本发明设计合成了32个迷迭香酸电子等排体衍生物,并对其进行酶水平和菌株水平的系统抗结核活性评价,迷迭香酸电子等排体衍生物及其盐或其溶剂化物为单一活性成分或作为活性成分之一,在抑制UGM 活性和抗牛型结核分枝杆菌的方面的应用,可制备成片剂、丸剂、胶囊剂、栓剂、乳剂、注射剂等药学上可接受的剂型,为抗结核创新药物的研发奠定基础。
2.采用本发明制备方法制备迷迭香酸生物电子等排体,产率高,在制备的32个迷迭香酸电子等排体衍生化合物中,其中4b,(N)-2-[(4-溴基)-苯丙酰胺基]-3-(4-联苯基)-丙酸产率达到了95.1%,产率达到80%以上的化合物为23个。
3.本发明32个化合物针对牛型结核分枝杆菌的体外筛选试验中发现,当化合物浓度在 500μg/mL时,有29个化合物对Mycobacterium Bovis(BCG)菌株有抑制作用。当化合物浓度在100μg/mL时,有14个化合物对Mycobacterium Bovis(BCG)菌株有抑制作用。
附图说明:
图1:化合物2a1H-NMR谱;
图2:化合物2a13C-NMR谱;
图3:化合物2a质谱图;
图4:化合物4a1H-NMR谱;
图5:化合物4a13C-NMR谱;
图6:化合物4a质谱图;
图7:化合物2a对小鼠巨噬细胞RAW264.7形态观察图;
图8:化合物4a对小鼠巨噬细胞RAW264.7形态观察图;
图9:化合物6a对小鼠巨噬细胞RAW264.7形态观察图;
图10:化合物7a对小鼠巨噬细胞RAW264.7形态观察图;
图11:化合物8a对小鼠巨噬细胞RAW264.7形态观察图;
图12:化合物4c对小鼠巨噬细胞RAW264.7形态观察图;
下面通过具体实施例,对本发明的技术方案作进一步地具体说明。
实施例1迷迭香酸生物电子等排体结构通式
实施例2迷迭香酸生物电子等排体化合物
符合结构通式1的衍生物为:
符合结构通式2的衍生物为:
实施例3迷迭香酸生物电子等排体化合物的制备
a.称取2.71g,10mmoL,1eq的O-苄基-L-酪氨酸于反应瓶中,向瓶内注入60mL 甲醇;将反应瓶置于冰盐浴下冷却20min;待体系温度降低至-10℃后,向反应瓶内缓慢滴加4.4mL,60mmoL,6eq重蒸的SOCl2,控制滴速,使体系温度保持在0℃以下;滴加完毕后,冰盐浴下反应10min,撤去冰盐浴后使反应瓶恢复至室温,再于80℃油浴下加热回流3h,TLC跟踪检测。反应结束后,减压回收溶剂,过滤,滤饼用二氯甲烷洗涤以获得所需化合物,将化合物真空抽干数小时后即得白色粉末状1;
b.称取5.26g,30mmoL,1eq的2,3-二氯苯甲醛于圆底烧瓶中,加入30mL吡啶并于100℃油浴中搅拌溶解样品;溶解后向反应瓶中加入9.36g,90mmoL,3eq的丙二酸并滴加880μL,9mmoL,0.3eq的哌啶于100℃油浴中加热回流3h后,TLC检测反应结束;反应体系冷却至室温后,将其倾入300mL冰水溶液中,用10%的盐酸调节pH值至4,有大量的白色固体析出;过滤,白色固体用蒸馏水洗3次;再用适量乙酸乙酯溶解白色固体,滤去不溶物;减压回收部分溶剂后置于冰箱中析出白色粉末状固体2;
c.称取0.44g,2mmoL,1eq的白色粉末状固体2置于反应瓶内,溶解于20mL无水DMF中,再加入0.58g,2mmoL,1eq的HBTU和695μL,4mmoL,2eq的DIEA,室温下反应30min;随后向反应物中加入0.57g,2mmoL,1eq的1,并于室温下反应 14h;TLC检测反应结束后,加入80mL乙酸乙酯和40mL蒸馏水分散,萃取,依次以蒸馏水和饱和氯化钠溶液洗涤,无水MgSO4干燥,减压回收乙酸乙酯至干所得粗品经比例为3:1的石油醚:乙酸乙酯硅胶柱层析纯化,得白色固体3;
d.取白色固体3,加入5mL蒸馏水和36mg,1.5mmoL,1.5eq的LiOH,室温搅拌反应2h,TLC检测反应结束;用1M浓盐酸调pH至2.5;减压回收溶剂至干,所得残留物再经二氯甲烷洗涤后过滤,抽干滤饼得化合物4a,(N)-2-[(5,6-二氯基)-苯丙烯酰胺基]-3-(4-苄氧基苯基)-丙酸。
实施例4迷迭香酸生物电子等排体化合物的制备
a.称取2.0g,10mmoL,1eq的O-苄基-L-酪氨酸于反应瓶中,向瓶内注入50mL甲醇;将反应瓶置于冰盐浴下冷却10min;待体系温度降低至-10℃后,向反应瓶内缓慢滴加4.4mL,60mmoL,6eq重蒸的SOCl2,控制滴速,使体系温度保持在0℃以下;滴加完毕后,冰盐浴下反应5min,撤去冰盐浴后使反应瓶恢复至室温,再于60℃油浴下加热回流2h,TLC跟踪检测。反应结束后,减压回收溶剂,过滤,滤饼用二氯甲烷洗涤以获得所需化合物,将化合物真空抽干数小时后即得白色粉末状1;
b.称取5.0g,30mmoL,1eq的2,3-二氯苯甲醛于圆底烧瓶中,加入20mL吡啶并于 80℃油浴中搅拌溶解样品;溶解后向反应瓶中加入9.0g,90mmoL,3eq的丙二酸并滴加880μL,9mmoL,0.3eq的哌啶于80℃油浴中加热回流2h后,TLC检测反应结束;反应体系冷却至室温后,将其倾入200mL冰水溶液中,用5%的盐酸调节pH值至4,有大量的白色固体析出;过滤,白色固体用蒸馏水洗2次;再用适量乙酸乙酯溶解白色固体,滤去不溶物;减压回收部分溶剂后置于冰箱中析出白色粉末状固体2;
c.称取0.4g,2mmoL,1eq的白色粉末状固体2置于反应瓶内,溶解于10mL无水 DMF中,再加入0.5g,2mmoL,1eq的HBTU和695μL,4mmoL,2eq的DIEA,室温下反应20min;随后向反应物中加入0.5g,2mmoL,1eq的1,并于室温下反应10h; TLC检测反应结束后,加入50mL乙酸乙酯和20mL蒸馏水分散,萃取,依次以蒸馏水和饱和氯化钠溶液洗涤,无水MgSO4干燥,减压回收乙酸乙酯至干所得粗品经比例为3:1的石油醚:乙酸乙酯硅胶柱层析纯化,得白色固体3;
d.取白色固体3,加入5mL蒸馏水和30mg,1.5mmoL,1.5eq的LiOH,室温搅拌反应1h,TLC检测反应结束;用1M浓盐酸调pH至2;减压回收溶剂至干,所得残留物再经二氯甲烷洗涤后过滤,抽干滤饼得化合物4a,(N)-2-[(5,6-二氯基)-苯丙烯酰胺基]-3-(4-苄氧基苯基)-丙酸。
实施例5迷迭香酸生物电子等排体化合物的制备
a.称取3.0g,10mmoL,1eq的O-苄基-L-酪氨酸于反应瓶中,向瓶内注入80mL甲醇;将反应瓶置于冰盐浴下冷却30min;待体系温度降低至-10℃后,向反应瓶内缓慢滴加4.4mL,60mmoL,6eq重蒸的SOCl2,控制滴速,使体系温度保持在0℃以下;滴加完毕后,冰盐浴下反应15min,撤去冰盐浴后使反应瓶恢复至室温,再于100℃油浴下加热回流5h,TLC跟踪检测。反应结束后,减压回收溶剂,过滤,滤饼用二氯甲烷洗涤以获得所需化合物,将化合物真空抽干数小时后即得白色粉末状1;
b.称取6.0g,30mmoL,1eq的2,3-二氯苯甲醛于圆底烧瓶中,加入50mL吡啶并于100℃油浴中搅拌溶解样品;溶解后向反应瓶中加入10.0g,90mmoL,3eq的丙二酸并滴加880μL,9mmoL,0.3eq的哌啶于80-100℃油浴中加热回流5h后,TLC检测反应结束;反应体系冷却至室温后,将其倾入500mL冰水溶液中,用15%的盐酸调节 pH值至4,有大量的白色固体析出;过滤,白色固体用蒸馏水洗3次;再用适量乙酸乙酯溶解白色固体,滤去不溶物;减压回收部分溶剂后置于冰箱中析出白色粉末状固体 2;
c.称取0.5g,2mmoL,1eq的白色粉末状固体2置于反应瓶内,溶解于30mL无水 DMF中,再加入0.6g,2mmoL,1eq的HBTU和695μL,4mmoL,2eq的DIEA,室温下反应50min;随后向反应物中加入0.6g,2mmoL,1eq的1,并于室温下反应1 5h; TLC检测反应结束后,加入100mL乙酸乙酯和50mL蒸馏水分散,萃取,依次以蒸馏水和饱和氯化钠溶液洗涤,无水MgSO4干燥,减压回收乙酸乙酯至干所得粗品经比例为3:1的石油醚:乙酸乙酯硅胶柱层析纯化,得白色固体3;
d.取白色固体3,加入10mL蒸馏水和40mg,1.5mmoL,1.5eq的LiOH,室温搅拌反应3h,TLC检测反应结束;用1M浓盐酸调pH至3;减压回收溶剂至干,所得残留物再经二氯甲烷洗涤后过滤,抽干滤饼得化合物4a,(N)-2-[(5,6-二氯基)-苯丙烯酰胺基]-3-(4-苄氧基苯基)-丙酸。
分别取实施例3-5任一制备的化合物,按常规制剂的制备方法,加入相关制剂辅料和赋型剂,分别制成片剂、膏剂、丸剂、胶囊剂、栓剂、乳剂和注射剂。
实施例6片剂
取化合物5g,加辅料羟丙甲纤维素6g、羧甲淀粉钠10g,微晶纤维素8g,乳糖 115g、淀粉50g,硬脂酸镁1g;将化合物与辅料充分混匀后投入高速搅拌机中,喷雾加水适量,整粒,水分控制在3~4%,然后压片,制成1000片,包薄膜衣。
为了进一步验证本发明的可行性及有效性,发明人进行了一系列的试验,具体如下:
1仪器及试剂
1.1仪器、型号及厂家
1.2主要原料及试剂
2实验例1
针对前期发现的UDP-半乳糖变异酶(UGM)抑制剂迷迭香酸,为改善其稳定性,通过生物电子等排策略,引入稳定性较好的酰胺键来重新构建该化合物。本发明设计合成了32个迷迭香酸电子等排体衍生物,并对其进行了酶水平和体外抗牛型结核分枝杆菌活性评价。本发明中涉及的化合物有较好的抗结核活性,说明本发明化合物可用于制备治疗结核感染的药物,有较潜在的临床应用价值。所述迷迭香酸电子等排体衍生物及其盐或其溶剂化物为单一活性成分或作为活性成分之一,可采用片剂、丸剂、胶囊剂、栓剂、乳剂和注射剂等药学上可接受的剂型。
2.1迷迭香酸电子等排体衍生物结构通式如下:
2.2迷迭香酸生物电子等排体为下列化合物,结构通式如下:
式1中,R1为相同或不同并且各自独立的氢、卤素、硝基、磺酸基、酰基、磺酰基、苯环、任意杂环或组合基团,组合基团是指,由酰基或磺酰基与苯环、任意杂环或链状脂肪烃的组合,且链状脂肪烃必须有除O以外的杂原子间断或为端基,且组合基团与苯环的连接方式为碳氮键或碳硫键;;且R1、R2不能全为卤素,若当R1、R2中任意一个为卤素时,则至少有另一个为除氢和卤素以外的上述基团;R1及R2为相同或不同且独立的含有O原子、N原子或S原子的芳香环、脂肪环或脂肪链,或不含O原子、N原子及S原子的芳香环或脂肪环,或包含上述芳香环、脂肪环及脂肪链的任意组合基团的烷基或酰基;包括C(2)的任何立体异构体。
式2中,R1为相同或不同并且各自独立的氢、卤素、硝基、磺酸基、酰基、磺酰基、苯环、任意杂环或组合基团,组合基团是指,由酰基或磺酰基与苯环、任意杂环或链状脂肪烃的组合,且链状脂肪烃必须有除O以外的杂原子间断或为端基,且组合基团与苯环的连接方式为碳氮键或碳硫键;且R1、R2不能全为卤素,若当R1、R2中任意一个为卤素时,则至少有另一个为除氢和卤素以外的上述基团;R1及R2为相同或不同且独立的含有O原子、N原子或S原子的芳香环、脂肪环或脂肪链,或不含O原子、N原子及S原子的芳香环或脂肪环,或包含上述芳香环、脂肪环及脂肪链的任意组合基团的烷基或酰基;包括C(2)的任何立体异构体。
符合结构通式1和2的衍生物,见表1、表2.
表1符合结构通式1的衍生物
表2符合结构通式2的衍生物
2.3制备方法
a.称取O-苄基-L-酪氨酸(2.71g,10mmoL,1eq)加入反应瓶中,向瓶内注入甲醇(60mL);将反应瓶置于冰盐浴下冷却20min;待体系温度降低至-10℃后,向反应瓶内缓慢滴加重蒸的SOCl2(4.4mL,60mmoL,6eq),控制滴速,使体系温度保持在 0℃以下;滴加完毕后,冰盐浴下反应10min,撤去冰盐浴后使反应瓶恢复至室温,再于80℃油浴下加热回流3h,TLC跟踪检测。反应结束后,减压回收溶剂,过滤,滤饼用二氯甲烷洗涤以获得所需化合物,将化合物真空抽干数小时后即得白色粉末状O- 苄基-L-酪氨酸甲酯盐酸盐1(1.549g,产率54%)。
b.称取2,3-二氯苯甲醛(5.26g,30mmoL,1eq)置于圆底烧瓶中,加入吡啶(30 mL)并于100℃油浴中搅拌溶解样品;溶解后向反应瓶中加入丙二酸(9.36g,90 mmoL,3eq)并滴加哌啶(880μL,9mmoL,0.3eq)于100℃油浴中加热回流3h后, TLC检测反应结束;反应体系冷却至室温后,将其倾入冰水溶液中(300mL),用10%的盐酸调节pH值至4,有大量的白色固体析出;过滤,白色固体用蒸馏水洗2-3 次;再用适量乙酸乙酯溶解白色固体,滤去不溶物;减压回收部分溶剂后置于冰箱中析出白色粉末状固体2(5.95g,产率91%)。
c.称取2(0.44g,2mmoL,1eq)置于反应瓶内,溶解于无水DMF(20mL)中,再加入HBTU(0.58g,2mmoL,1eq)和DIEA(695μL,4mmoL,2eq),室温下反应 30min;随后向反应物中加入1(0.57g,2mmoL,1eq),并于室温下反应14h;TLC 检测反应结束后,加入乙酸乙酯(80mL)和蒸馏水(40mL)分散,萃取,依次以蒸馏水和饱和氯化钠溶液洗涤,无水MgSO4干燥,减压回收乙酸乙酯至干所得粗品经硅胶柱层析[V(石油醚):V(乙酸乙酯)=3:1]纯化,得白色固体3(0.51g,产率53.1%)
d.溶解后,加入蒸馏水(5mL)和LiOH(36mg,1.5mmoL,1.5eq),室温搅拌反应2h,TLC检测反应结束;用浓盐酸(1M)调pH至2-3;减压回收溶剂至干,所得残留物4a,即(N)-2-[(5,6-二氯基)-苯丙烯酰胺基]-3-(4-苄氧基苯基)-丙酸,白色固体,产率:90.6%。
ESI-MS:m/z 468.0766[M-H]+;1H-NMR(CD3OD,600MHz)δ(ppm):2.98(m,1H,3- Ha),3.21(m,1H,3-Hb),4.77(m,1H,2-CH),5.03(s,2H,1″-CH2),6.67(d,J=15.7Hz,1H, 2′-CH)6.91(d,J=8.6Hz,2H,6,8-ArH),7.16(d,J=8.6Hz,2H,5,9-ArH),7.23-7.43(m, 6H,8′,9′,3″,4″,5″,6″-ArH),7.58(m,2H,7′,7″-ArH),7.90(d,J=1.8Hz,1H,3′-CH).13C- NMR(CD3OD,150MHz)δ(ppm):37.7(C-3),55.5(C-2),71.0(C-1″),115.9(C-6,8),127.1 (C-2′),128.7(C-9′),128.8(C-3″,7″),129.1(C-5′,8′,5″),129.7(C-4,4″,6″),130.9(C-7′), 133.5(C-5,9),134.7(C-4′),136.8(C-2″),137.7(C-6′),138.8(C-3′),159.2(C-7),167.4(C- 1′),174.5(C-1′).
用相同的方法可制得32个迷迭香酸生物电子等排体。
(1)1a,(N)-2-[3-(3,4-二氯苯基)-苯丙烯酰胺基]-3-(4-硝基苯基)-丙酸,淡黄色固体,产率:57.2%;ESI-MS:m/z 407.0197[M-H]+;1H-NMR(CD3OD,600MHz)δ(ppm): 3.13(m,1H,3-Ha),3.36(m,1H,3-Hb),4.87(m,1H,2-CH),6.72(d,J=12Hz,1H,2′-CH). 13C-NMR(CD3OD,150MHz)δ(ppm):38.4(C-3),55.6(C-2),126.0(C-6),127.4(C-8), 128.2(C-2′),128.3(C-9′),128.5(C-5′),129.3(C-5),130.1(C-9),131.0(C-8′),133.8(C-7′), 135.0(C-6′),137.1(C-4′),137.7(C-3′),138.0(C-4),141.4(C-7),167.8(C-1′),174.8(C-1);
(2)2a,(N)-2-[(3,4-二氯基)-苯丙烯酰胺基]-3-(4-苄氧基苯基)-丙酸,白色固体,产率:57.9%;ESI-MS:m/z 494.0965[M-H]+;1H-NMR(CD3OD,600MHz)δ(ppm):2.97 (m,1H,3-Ha),3.20(m,1H,3-Hb),4.75(m,1H,2-CH),5.03(s,2H,1″-CH2),6.66(d,J=8.6 Hz,1H,2′-CH)6.90(d,J=8.6Hz,2H,6,8-ArH),7.16(m,2H,5,9-ArH),7.23-7.57(m,8H, 6′,8′,9′,3″,4″,5″,6″,7″-ArH),7.71(d,J=1.8Hz,1H,5′-ArH).13C-NMR(CD3OD,150 MHz)δ(ppm):38.0(C-3),55.8(C-2),71.3(C-1″),116.2(C-6,8),123.9(C-2′),128.7(C-9′, 3″,7″),128.8(C-5″),129.1(C-5′),129.7(C-4,4″,6″),130.9(C-5,9,8″),131.6(C-7′),132.3 (C-6′),134.6(C-4′),137.1(C-2″),139.8(C-3′),159.5(C-7),167.9(C-1′),174.9(C-1);见图 1、图2、图3。
(3)3a,(N)-2-[(5,6-二氯基)-苯丙烯酰胺基]-3-(4-硝基苯基)-丙酸,淡黄色固体,产率:87.0%;ESI-MS:m/z 407.0196[M-H]+;1H-NMR(CD3OD,600MHz)δ(ppm):3.10 (m,1H,3-Ha),3.28(m,1H,3-Hb),468(m,1H,2-CH),6.73(d,J=15.7Hz,1H,2′-CH), 7.39-7.70(m,6H,3′-CH,5,9,7′,8′,9′-ArH),8.16(d,J=8.7Hz,2H,6,8-ArH).13C-NMR (CD3OD,150MHz)δ(ppm):36.0(C-3),52.6(C-2),122.8(C-6,8),125.4(C-9′),125.8(C-5′, 8′),128.0(C-7′),130.0(C-9),130.7(C-5),132.0(C-4′),134.2(C-6′),134.6(C-4),145.4(C-3′),145.9(C-7),163.7(C-1′),171.8(C-1);
(4)4a,(N)-2-[(5,6-二氯基)-苯丙烯酰胺基]-3-(4-苄氧基苯基)-丙酸,白色固体,产率:90.6%;ESI-MS:m/z 468.0766[M-H]+;1H-NMR(CD3OD,600MHz)δ(ppm):2.98 (m,1H,3-Ha),3.21(m,1H,3-Hb),4.77(m,1H,2-CH),5.03(s,2H,1″-CH2),6.67(d,J= 15.7Hz,1H,2′-CH)6.91(d,J=8.6Hz,2H,6,8-ArH),7.16(d,J=8.6Hz,2H,5,9-ArH), 7.23-7.43(m,6H,8′,9′,3″,4″,5″,6″-ArH),7.58(m,2H,7′,7″-ArH),7.90(d,J=1.8Hz,1H, 3′-CH).13C-NMR(CD3OD,150MHz)δ(ppm):37.7(C-3),55.5(C-2),71.0(C-1″),115.9 (C-6,8),127.1(C-2′),128.7(C-9′),128.8(C-3″,7″),129.1(C-5′,8′,5″),129.7(C-4,4″,6″),130.9(C-7′),133.5(C-5,9),134.7(C-4′),136.8(C-2″),137.7(C-6′),138.8(C-3′),159.2(C- 7),167.4(C-1′),174.5(C-1′);见图4、图5、图6。
(5)5a,(N)-2-[(5,6-二氯基)-苯丙烯酰胺基]-3-(4-碘基苯基)-丙酸,白色固体,产率:87.8%;ESI-MS:m/z 487.9312[M-H]+;1H-NMR(CD3OD,600MHz)δ(ppm):3.01 (m,1H,3-Ha),3.23(m,1H,3-Hb),4.80(m,1H,2-CH),6.66(d,J=15.7Hz,1H,2′-CH), 7.04(d,J=8.4Hz,2H,5,9,-ArH),7.31(m,1H,5′-ArH),7.54(m,1H,6′-ArH),7.63(m,3H, 6,8,7′-ArH),7.9(d,J=15.6,1H,3′-CH).13C-NMR(CD3OD,150MHz)δ(ppm):38.0(C-3), 55.0(C-2),125.5(C-6,8),127.1(C-9′),129.0(C-5′,8′),132.3(C-7′),132.5(C-9),133.5(C- 5),134.7(C-4′),136.8(C-6′),137.9(C-4),138.2(C-3′),138.7(C-7),167.4(C-1′),174.1(C- 1);
(6)6a,(N)-2-[(5,6-二氯基)-苯丙烯酰胺基]-3-(4-联苯基)-丙酸,白色固体,产率:81.6%;ESI-MS:m/z 462.0634[M+Na]+;1H-NMR(DMSO-d6,600MHz)δ(ppm): 3.00(m,1H,3-Ha),3.18(m,1H,3-Hb),4.62(m,1H,2-CH),6.79(d,J=15.7Hz,1H,2′-CH), 7.27-7.73(m,13H,3′-CH,5,6,8,9,5′,6′,7′,2″,3″,4″,5″,6″-ArH);
(7)7a,(N)-2-[(3,4-二氯基)-苯丙烯酰胺基]-3-(4-联苯基)-丙酸,白色固体,产率:92.9%;ESI-MS:m/z 438.0658[M-H]+;1H-NMR(DMSO-d6,600MHz)δ(ppm):2.99 (m,1H,3-Ha),3.17(m,1H,3-Hb),4.62(m,1H,2-CH),6.79(d,J=15.9Hz,1H,2′-CH), 7.29-7.49(m,6H,5,6,8,9,5′,8′-ArH),7.52-7.70(m,6H,3′-CH,2″,3″,4″,5″,6″-ArH), 7.84(d,J=1.8Hz,1H,9′-ArH).13C-NMR(DMSO-d6,150MHz)δ(ppm):36.2(C-3),53.4 (C-2),123.8(C-2′),126.4(C-9′),127.2(C-5′,2″,4″,6″),127.3(C-3″,5″),128.8(C-5,9), 129.4(C-8′),131.0(C-7′),131.5(C-6′),131.6(C-4′),135.6(C-4),136.6(C-7),138.2(C-1″),139.8(C-3′),164.3(C-1′),172.7(C-1);
(8)8a,(N)-2-[(3,4-二氯基)-苯丙烯酰胺基]-3-(4-碘基苯基)-丙酸,白色固体,产率:70.3%;ESI-MS:m/z 487.9312[M-H]+;1H-NMR(CD3OD,600MHz)δ(ppm):3.00(m, 1H,3-Ha),3.22(m,1H,3-Hb),4.79(m,1H,2-CH),6.65(d,J=15.8Hz,1H,2′-CH),7.03(d, J=8.2Hz,2H,5,9,-ArH),7.36-7.86(m,6H,3′-CH,6,8,5′,6′,9′-ArH);
(9)9a,(N)-2-苯丙烯酰胺基-3-(4-硝基苯基)-丙酸,棕黄色固体,产率:79.9%;ESI-MS:m/z 363[M+Na]+;1H-NMR(DMSO-d6,600MHz)δ(ppm):3.09(m,1H,3-Ha), 3.28(m,1H,3-Hb),4.67(m,1H,2-CH),6.68(d,J=15.8Hz,1H,2′-CH),7.31-7.45(m,4H, 3′-CH,6′,7′,8′-ArH),,7.54-7.56(m,4H,5′,9′,5,9-ArH),8.15(m,2H,6,8-ArH).13C- NMR(DMSO-d6,150MHz)δ(ppm):36.5(C-3),53.0(C-2),121.5(C-6,8),123.3(C-2′), 127.6(C-7′),129.0(C-5′,9′),130.5(C-6′,8′),134.7(C-5,9),139.4(C-4′),146.0(C-3′),146.4 (C-4),165.0(C-1′),172.5(C-1);
(10)10a,(N)-2-苯丙烯酰胺基-3-(4-溴-苯基)-丙酸,白色固体,产率:26%;ESI-MS:m/z 396[M+Na]+;1H-NMR(DMSO-d6,600MHz)δ(ppm):2.92(m,1H,3-Ha),3.10 (m,1H,3-Hb),4.58(m,1H,2-CH),6.69(d,J=15.8Hz,1H,2′-CH),7.21(d,2H,5,9,-ArH), 7.34-7.43(m,6H,3′-CH,6′,8′,5′,7′,9′-ArH),7.55(d,2H,6,8-ArH).13C-NMR(DMSO-d6, 150MHz)δ(ppm):36.2(C-3),55.4(C-2),119.7(C-7),121.6(C-2′),127.6(C-7′),129.0(C- 5′,9′),131.0(C-6′,8′),131.4(C-5,6,8,9),134.8(C-4′),137.0(C-4),139.3(C-3′),164.9(C-1′),172.8(C-1);
(11)11a,(N)-2-苯丙烯酰胺基-3-苯丙酸,淡黄色固体,产率:70.9%;ESI-MS: m/z 318[M+Na]+;1H-NMR(DMSO-d6,600MHz)δ(ppm):2.99(m,1H,3-Ha),3.12(m, 1H,3-Hb),4.5(m,1H,2-CH),6.73(d,J=15.8Hz,1H,2′-CH),7.19(d,J=15.6Hz,1H,3′- CH),7.21-7.27(m,4H,5,7,9,7′-H),7.33-7.45(m,4H,6,8,6′,8′-H),7.54(d,2H,5′,9′- ArH).13C-NMR(DMSO-d6,150MHz)δ(ppm):36.7(C-3),53.6(C-2),121.6(C-2′),126.3 (C-7),127.5(C-5,9),128.1(C-7′),128.8(C-5′,9′),129.0(C-6′,8′),129.5(C-6,8),134.7(C- 4′),137.6(C-4),139.0(C-3′),164.8(C-1′),172.9(C-1);
(12)12a,(N)-2-苯丙烯酰胺基-3-(4-碘苯基)-丙酸,白色固体,产率:32.7%;ESI-MS:m/z 330.0[M+Na]+;1H-NMR(DMSO-d6,600MHz)δ(ppm):2.89(m,1H,3-Ha), 3.08(m,1H,3-Hb),4.53(m,1H,2-CH),6.69(d,J=18Hz,1H,2′-CH),7.07(d,2H,6,8- ArH),7.36-7.42(m,4H,3′-CH,5,9,7′-ArH),7.55(d,J=12Hz,6′,8′-ArH),7.64(d,2H,5′, 9′-ArH).13C-NMR(DMSO-d6,150MHz)δ(ppm):36.3(C-3),53.4(C-2),92.5(C-7),121.6 (C-2′),127.6(C-7′),129.0(C-5′,9′),129.6(C-6′,8′),131.6(C-5,9),134.8(C-4),137.0(C-4′), 137.4(C-6,8),139.4(C-3′),164.9(C-1′),172.9(C-1);
(13)13a,(N)-2-苯丙烯酰胺基-3-(4-氯苯基)-丙酸,白色固体,产率:38.4%;ESI-MS:m/z 444.0[M+Na]+;1H-NMR(DMSO-d6,600MHz)δ(ppm):2.88(m,1H,3-Ha), 3.09(m,1H,3-Hb),4.51(m,1H,2-CH),6.62(d,J=18Hz,1H,2′-CH),7.03(m,8H,2′-CH, 5,9,5′,6′,7′,8′,9′-ArH),7.53(d,2H,6,8-ArH).13C-NMR(DMSO-d6,150MHz)δ(ppm): 36.7(C-3),54.4(C-2),121.7(C-2′),128.3(C-7′),128.8(C-5,9),129.6(C-5′,6′,8′,9′),130.4, (C-6,8),131.6(C-7),135.0(C-4′),137.1(C-4),140.4(C-3′),165.9(C-1′),173.5(C-1);
(14)14a,(N)-2-苯丙烯酰胺基-3-(4-羟基苯基)-丙酸,白色固体,产率:28.1%;ESI-MS:m/z 334[M+Na]+;1H-NMR(DMSO-d6,600MHz)δ(ppm):2.81(m,1H,3-Ha), 3.00(m,1H,3-Hb),4.49(m,1H,2-CH),6.69(m,3H,2′-CH,6,8-ArH),7.04(m,2H,5,9- ArH),7.35-7.49(m,4H,6′,7′,8′-ArH,3′-CH),7.55(m,2H,5′,9′-ArH).13C-NMR(DMSO- d6,150MHz)δ(ppm):36.1(C-3),54.0(C-2),115.0(C-7),121.8(C-2′),127.6(C-5,9), 129.0(C-5′,6′,8′,9′),130.0(C-7′),134.8(C-4,4′),139.2(C-6,8),164.9(C-3′),170.2(C-1′), 173.1(C-1);
(15)1b,(N)-2-[(4-溴基)-苯丙酰胺基]-3-(4-硝基苯基)-丙酸,淡黄色固体,产率: 78.4%;ESI-MS:m/z 443.0219[M+Na]+;1H-NMR(CD3OD,600MHz)δ(ppm):2.54-2.41 (m,2H,2′-CH2),2.80(m,2H,3′-CH2),3.05(m,1H,3-Ha),3.27(m,1H,3-Hb),4.73(d,J= 12Hz,1H,2-CH),7.07(d,J=6Hz,2H,5′,9′-ArH),7.34-7.37(m,4H,6′,8′,5,9-ArH),8.11 (d,2H,6,8-ArH).13C-NMR(CD3OD,150MHz)δ(ppm):31.9(C-3′),38.1(C-2′),38.2(C-3), 54.5(C-2),120.9(C-7′),124.5(C-6,8),131.5(C-5,9),132.6(C-5′,6′,8′,9′),141.5(C-4′),146.7(C-4),148.5(C-7),174.1(C-1′),174.9(C-1);
(16)2b,(N)-2-[(4-溴基)-苯丙酰胺基]-3-(4-苄氧基苯基)-丙酸,白色固体,产率: 81.3%;ESI-MS:m/z 504.0784[M+Na]+;1H-NMR(CD3OD,600MHz)δ(ppm):2.45(m, 2H,2′-CH2),2.78(m,2H,3′-CH2),2.85(m,1H,3-Ha),3.09(m,1H,3-Hb),4.60(m,1H,2- CH),5.04(s,2H,1″-CH2),6.88(d,2H,J=8.5Hz,6,8-ArH),7.04(m,4H,5,9,5′,9′-ArH), 7.28-7.41(m,7H,6′,8′,3″,4″,5″,6″,7″-ArH).13C-NMR(CD3OD,150MHz)δ(ppm):32.2 (C-3′),37.8(C-2′),38.4(C-3),55.4(C-2),71.2(C-1″),116.2(C-6,8),121.1(C-7′),128.8(C-3″,7″),129.1(C-5″),129.8(C-4′,4″,6″),130.9(C-5,9),131.6(C-5′,6′,8′,9′),132.8(C-2″), 139.1(C-4′),141.8(C-4),159.4(C-7),175.0(C-1′);
(17)3b,(N)-2-[(4-溴基)-苯丙酰胺基]-3-(4-碘苯基)-丙酸,白色固体,产率:87.7%;ESI-MS:m/z 523.9328[M+Na]+;1H-NMR(CD3OD,600MHz)δ(ppm):2.46(m, 2H,2′-CH2),2.80(m,2H,3′-CH2),2.87(m,1H,3-Ha),3.11(m,1H,3-Hb),4.64(d,J=14Hz, 1H,2-CH),6.90(d,J=8.2Hz,2H,5,9-ArH),7.06(d,J=8.3Hz,2H,5′,9′-ArH),7.38(d,J =8.3Hz,2H,6,8-ArH),7.59(d,J=8.2Hz,2H,6′,8′-ArH);
(18)4b,(N)-2-[(4-溴基)-苯丙酰胺基]-3-(4-联苯基)-丙酸,白色固体,产率:95.1%;ESI-MS:m/z 474.0676[M+Na]+;1H-NMR(CD3OD,600MHz)δ(ppm):2.39(m, 2H,2′-CH2),2.72(m,2H,3′-CH2),2.89(m,1H,3-Ha),3.07(m,1H,3-Hb),4.46(m,1H,2- CH),7.10(d,J=8.3Hz,2H,5′,9′-ArH),7.25(d,J=8.1Hz,2H,5,9-ArH),7.30-7.50(m, 5H,6,8,3″,4″,5″-ArH),7.56(d,J=8.1Hz,2H,2″,6″-ArH).7.64(d,J=7.3Hz,2H,6′,8′- ArH).13C-NMR(CD3OD,150MHz)δ(ppm):25.1(C-3′),30.1(C-2′),36.2(C-3),53.4(C-2), 118.9(C-7′),126.4(C-6,8),127.3(C-2″),129.0(C-4″,6″),129.7(C-3″,5″),130.5(C-5,9),131.0(C-5′,6′,8′,9′),137.0(C-4),138.3(C-7),139.9(C-4′),140.7(C-1″),171.2(C-1′), 173.0(C-1);
(19)5b,(N)-2-[(4-氯基)-苯丙酰胺基]-3-(4-苄氧基苯基)-丙酸,白色固体,产率:92.0%;ESI-MS:m/z 460.1288[M+Na]+;1H-NMR(CD3OD,600MHz)δ(ppm):2.45 (m,2H,2′-CH2),2.79(m,2H,3′-CH2),2.85(m,1H,3-Ha),3.09(m,1H,3-Hb),4.61(m,1H, 2-CH),5.04(s,2H,1″-CH2),6.88(d,J=8.5Hz,2H,6,8-ArH),7.03-7.11(m,4H,5,9,5′,9′- ArH),7.21-7.29(m,3H,4″,5″,6″-ArH),7.34(m,2H,6′,8′-ArH),7.41(d,J=7.6Hz,2H,3″, 7″-ArH).13C-NMR(CD3OD,150MHz)δ(ppm):31.9(C-3′),37.6(C-2′),38.2(C-3),55.1 (C-2),71.0(C-1″),115.9(C-6,8),128.5(C-7′),128.8(C-3″,7″),129.5(C-5″),130.7(C-4′,4″,6″),131.0(C-5,9),131.2(C-5′,6′,8′,9′),132.9(C-2″),138.8(C-4′),141.0(C-4),159.2 (C-7),174.8(C-1,1′);
(20)6b,(N)-2-[(4-氯基)-苯丙酰胺基]-3-(4-硝基苯基)-丙酸,淡黄色固体,产率:74.5%;ESI-MS:m/z.399.0722[M+Na]+;1H-NMR(CD3OD,600MHz)δ(ppm): 2.52-2.41(m,2H,2′-CH2),2.82(m,2H,3′-CH2),3.04(m,1H,3-Ha),3.28(m,1H,3-Hb), 4.72(m,1H,2-CH),7.12-7.21(m,4H,5′,6′,8′9′-ArH),7.35(m,2H,5,9-ArH),8.11(d,2H, 6,8-ArH).13C-NMR(CD3OD,150MHz)δ(ppm):31.8(C-3′),38.0(C-2′),38.1(C-3),54.3 (C-2),124.4(C-7′),129.4(C-6,8),131.0(C-5,9),131.4(C-6′,8′),133.0(C-5′,9′),140.8(C-4′), 146.7(C-4),148.3(C-7),174.0(C-1′),174.8(C-1);
(21)7b,(N)-2-[(4-氯基)-苯丙酰胺基]-3-(4-碘基苯基)-丙酸,白色固体,产率:88.6%;ESI-MS:m/z 455.9863[M+Na]+;1H-NMR(CD3OD,600MHz)δ(ppm):2.46(m, 2H,2′-CH2),2.80(m,2H,3′-CH2),2.87(m,1H,3-Ha),3.11(m,1H,3-Hb),4.59(m,1H,2- CH),6.90(d,J=8.2Hz,2H,5,9-ArH),7.12(d,J=8.3Hz,2H,5′,9′-ArH),7.23(d,J=8.3Hz, 2H,6,8-ArH),7.57(d,J=8.2Hz,2H,6′,8′-ArH).13C-NMR(CD3OD,150MHz)δ(ppm): 31.8(C-3′),38.1(C-2′),38.3(C-3),55.3(C-2),92.5(C-7),129.5(C-6′,8′),131.0(C-5′,9′), 132.5(C-5,9),132.9(C-7′),138.5(C-4),138.6(C-6,8),140.9(C-4′),174.5(C-1,1′);
(22)8b,(N)-2-[(4-氯基)-苯丙酰胺基]-3-(4-联苯基)-丙酸,白色固体,产率:91.7%;ESI-MS:m/z 406.1027[M-H]+;1H-NMR(CD3OD,600MHz)δ(ppm):2.48(m, 2H,2′-CH2),2.81(m,2H,3′-CH2),2.97(m,1H,3-Ha),3.21(m,1H,3-Hb),4.70(m,1H,2- CH),7.11(d,J=8.4Hz,2H,5′,9′-ArH),7.20(m,4H,5,9,6′,8′-ArH),7.36(m,3H,6,8,4″- ArH),7.54(m,4H,2″,3″,5″,6″-ArH).13C-NMR(CD3OD,150MHz)δ(ppm):31.9(C-3′), 38.0(C-2′),38.2(C-3),55.0(C-2),127.9(C-7′),128.0(C-6,8),128.3(C-2″),129.5(C-4″, 6″),129.8,(C-3″,5″),130.7(C-5,9),131.0(C-5′,6′,8′,9′),132.9(C-4),137.6(C-7),141.0(C-4′),142.1(C-1″),174.5(C-1′),174.8(C-1);
(23)9b,(N)-2-[(4-氟基)-苯丙酰胺基]-3-(4-苄氧基苯基)-丙酸白色固体,产率: 92.0%;ESI-MS:m/z 420.1611[M-H]+;1H-NMR(CD3OD,600MHz)δ(ppm):2.44(m, 2H,2′-CH2),2.79(m,2H,3′-CH2),2.85(m,1H,3-Ha),3.10(m,1H,3-Hb),4.60(m,1H,2- CH),5.04(s,2H,1″-CH2),6.88(d,J=8.5Hz,2H,6,8-ArH),6.94(m,2H,5,9-ArH),7.06(d, J=8.9Hz,,2H,6′,8′-ArH),7.12(m,2H,5′,9′-ArH),7.27-7.41(m,5H,3″,4″,5″,6″,7″-ArH).13C-NMR(CD3OD,150MHz)δ(ppm):32.1(C-3′),37.9(C-2′),38.8(C-3),55.4(C-2),71.2(C-1″),116.1(C-6),116.3(C-8),128.5(C-7′),128.8(C-3″,7″),129.5(C-5″),130.7(C-4′, 4″,6″),131.0(C-5,9),131.2(C-5′,6′,8′,9′),132.9(C-2″),138.8(C-4′),162.3(C-4),163.9 (C-7),175.1(C-1′),175.2(C-1);
(24)10b,(N)-2-[(4-氟基)-苯丙酰胺基]-3-(4-硝基苯基)-丙酸,淡黄色固体,产率:81.2%;ESI-MS:m/z 359.1041[M-H]+;1H-NMR(CD3OD,600MHz)δ(ppm):2.46 (m,2H,2′-CH2),2.81(m,2H,3′-CH2),3.04(m,1H,3-Ha),3.29(m,1H,3-Hb),4.72(d,J= 12Hz,1H,2-CH),6.92(m,2H,5′,9′-ArH),7.13(m,2H,6′,8′-ArH),7.37(d,J=8.6Hz,2H, 5,9-ArH),8.11(d,J=8.7Hz,2H,6,8-ArH).13C-NMR(CD3OD,150MHz)δ(ppm):31.9 (C-3′),38.0(C-2′),38.2(C-3),55.0(C-2),127.9(C-7′),128.0(C-6,8),128.3(C-2″),129.5 (C-4″,6″),129.8,(C-3″,5″),130.7(C-5,9),131.0(C-5′,6′,8′,9′),132.9(C-4),137.6(C-7),141.0(C-4′),142.1(C-1″),174.5(C-1′),174.8(C-1);
(25)11b,(N)-2-[(4-氟基)苯丙酰胺基]-3-(4-碘基苯基)-丙酸,白色固体,产率:88.6%;ESI-MS:m/z 440.0157[M-H]+;1H-NMR(CD3OD,600MHz)δ(ppm):2.45(m, 2H,2′-CH2),2.80(m,2H,3′-CH2),2.86(m,1H,3-Ha),3.12(m,1H,3-Hb),4.64(m,1H,2- CH),6.94(m,4H,5,9,5′,9′-ArH),7.13(m,2H,6,8-ArH),7.59(m,2H,6′,8′-ArH).13C- NMR(CD3OD,150MHz)δ(ppm):31.7(C-3′),38.1(C-2′),38.4(C-3),54.3(C-2),115.9(C- 7′),116.0(C-6,8),124.4(C-5,9),131.0(C-5′),131.4(C-6′),138.0(C-8′),146.6(C-9′),148.3 (C-4′),162.0(C-4),163.7(C-7),174.0(C-1′),174.9(C-1);
(26)12b,(N)-2-[(4-氟基)-苯丙酰胺基]-3-(4-联苯基)-丙酸,白色固体,产率:91.7%;ESI-MS:m/z 390.1505[M-H]+;1H-NMR(CD3OD,600MHz)δ(ppm):2.47(m,2H, 2′-CH2),2.81(m,2H,3′-CH2),2.97(m,1H,3-Ha),3.21(m,1H,3-Hb),4.70(m,1H,2-CH),, 6.92(m,2H,6′,8′-ArH),7.13(m,2H,5′,9′-ArH),7.23(m,2H,5,9-ArH),7.36(m,3H,6,8, 4″-ArH),7.54(m,4H,2″,3″,5″,6″-ArH).13C-NMR(CD3OD,150MHz)δ(ppm):31.8(C- 3′),38.0(C-2′),38.5(C-3),54.9(C-2),116.0(C-,6′,8′),128.0(C-6,8),128.3(C-2″,4″,6″),129.8,(C-3″,5″),130.8(C-5,9),131.0(C-5′,9′),137.6(C-4),138.0(C-4′),141.0(C-7), 142.1(C-1″),162.0(C-7′)174.7(C-1′),175.0(C-1);
(27)13b,(N)-2-[(4-碘基)-苯丙酰胺基]-3-(4-苄氧基苯基)-丙酸,白色固体,产率:88.5%;ESI-MS:m/z 552.0647[M-H]+;1H-NMR(CD3OD,600MHz)δ(ppm):2.44 (m,2H,2′-CH2),2.77(m,2H,3′-CH2),2.85(m,1H,3-Ha),3.09(m,1H,3-Hb),4.60(m,1H, 2-CH),5.05(s,2H,1″-CH2),6.90(m,4H,6,8,5′,9′-ArH),7.03(m,2H,5,9-ArH),7.27-7.42 (m,5H,3″,4″,5″,6″,7″-ArH),7.56(d,J=8.1Hz,,2H,6′,8′-ArH).13C-NMR(CD3OD,150 MHz)δ(ppm):32.4(C-3′),38.0(C-2′),38.5(C-3),55.5(C-2),71.4(C-1″),116.3(C-6,8), 129.0(C-7′),129.2(C-3″,7″),129.9(C-5″),131.0(C-4′,4″,6″),131.7(C-5,9),132.1(C-5′, 6′,8′,9′),139.0(C-2″),139.2(C-4′),142.4(C-4),159.5(C-7),175.1(C-1′),175.2(C-1);
(28)14b,(N)-2-[(4-碘基)-苯丙酰胺基]-3-(4-硝基苯基)-丙酸,淡黄色固体,80.1%;ESI-MS:m/z 491.0080[M+Na]+;1H-NMR(CD3OD,600MHz)δ(ppm):2.47(m, 2H,2′-CH2),2.79(m,2H,3′-CH2),3.05(m,1H,3-Ha),3.28(m,1H,3-Hb),4.72(m,1H,2- CH),6.94(d,J=8.2Hz,,2H,5′,9′-ArH),7.34(d,J=8.6Hz,2H,5,9-ArH),7.56(d,J=8.2 Hz,2H,6′,8′-ArH),8.12(d,J=8.6Hz,2H,6,8-ArH).13C-NMR(CD3OD,150MHz)δ (ppm):32.2(C-3′),38.2(C-2′),38.4(C-3),54.6(C-2),92.1(C-7′),124.7(C-6,8),131.7(C-5, 9′),132.0(C-5,9),138.9(C-6′,8′),142.2(C-4′),146.8(C-4),148.6(C-7),174.2(C-1′),175.0(C-1);
(29)15b,(N)-2-[(4-碘基)-苯丙酰胺基]-3-(4-联苯基)-丙酸白色固体,产率:87.5%;ESI-MS:m/z 522.0541[M+Na]+;1H-NMR(CD3OD,600MHz)δ(ppm):2.38(m, 2H,2′-CH2),2.70(m,2H,3′-CH2),2.88(m,1H,3-Ha),3.07(m,1H,3-Hb),4.45(m,1H,2- CH),6.96(d,J=8.2Hz,2H,5′,9′-ArH),7.24(d,J=8.1Hz,2H,5,9-ArH),7.35(m,1H,4″- ArH),7.45(m,2H,6,8-ArH),7.56(m,4H,2″,3″,5″,6″-ArH),7.64(d,J=7.4Hz,2H,6′, 8′-ArH).13C-NMR(CD3OD,150MHz)δ(ppm):29.7(C-3′),35.6(C-2′),35.8(C-3),52.8(C- 2),90.9(C-7′),125.9(C-5′,9′),126.0(C-6,8,2″,4″,6″),128.4(C-3″,5″),129.2(C-5,9), 130.2(C-4),136.4(C-6′,8′),137.7(C-7),139.4(C-4′),140.5(C-1″),170.7(C-1′),172.5(C-1);
(30)16b,(N)-2-[(4-碘基)-苯丙酰胺基]-3-(4-碘基苯基)-丙酸,白色固体,产率: 87.3%;ESI-MS:m/z 571.9193[M-H]+;1H-NMR(CD3OD,600MHz)δ(ppm):2.35(m,2H, 2′-CH2),2.68(m,2H,3′-CH2),2.78(m,1H,3-Ha),2.98(m,1H,3-Hb),4.3(m,1H,2-CH), 6.94(m,4H,5,9,5′,9′-ArH),7.59(m,4H,6,8,6′,8′-ArH).13C-NMR(CD3OD,150MHz)δ (ppm):29.7(C-3′),35.6(C-2′),35.7(C-3),56.6(C-2),91.0(C-7′),91.8(C-7),130.2(C-5′,9′),131.0(C-5,9),136.4(C-4),136.4(C-6′,8′),137.0(C-6,8),140.5(C-4′),170.6(C-1′),172.3 (C-1);
(31)17b,(N)-2-[(4-氰基)-苯丙酰胺基]-3-(4-苄氧基苯基)-丙酸,白色固体,产率:91.1%;ESI-MS:m/z 427.1655[M-H]+;1H-NMR(CD3OD,600MHz)δ(ppm):2.49 (m,2H,2′-CH2),2.84(m,1H,3-Ha),2.89(m,2H,3′-CH2),3.11(m,1H,3-Hb),4.60(m,1H, 2-CH),5.05(s,2H,1″-CH2),6.88(m,2H,6,8-ArH),7.06(m,2H,5,9-ArH),7.27-7.35(m, 5H,3″,4″,5″,6″,7″-ArH),7.42(d,J=7.5Hz,,2H,5′,9′-ArH),7.57(d,J=8.2Hz,,2H,6′, 8′-ArH);
(32)18b,(N)-2-[(4-氰基)-苯丙酰胺基]-3-(4-硝基苯基)-丙酸,淡黄色固体,产率:83.4%;ESI-MS:m/z 366.1087[M-H]+;1H-NMR(CD3OD,600MHz)δ(ppm):2.51 (m,2H,2′-CH2),2.91(m,2H,3′-CH2),3.04(m,1H,3-Ha),3.30(m,1H,3-Hb),4.73(m,1H, 2-CH),7.37(m,4H,5,9,5′,9′-ArH),7.58(d,J=8.2Hz,2H,6′,8′-ArH),8.12(d,J=8.6Hz, 2H,6,8-ArH).13C-NMR(CD3OD,150MHz)δ(ppm):32.4(C-3′),38.2(C-2′),38.2(C-3), 54.3(C-2),111.0(C-7′),119.8(C-6,8),124.4(C-5′,9′),130.6(C-5,9),131.4(C-6′,8′),133.3 (C-4′),146.6(C-4),148.2(C-7),174.0(C-1′),174.3(C-1)。
2.4抑制活性实验
在对上述32个化合物进行荧光偏振法测定亲和力测定,每个测试样品配置为11个梯度浓度供试品(2000μM、1000μM、500μM、100μM、20μM、10μM、4μM、 1μM、0.1μM、0.01μM、0μM),每个浓度吸取20μL加入384微孔板中,随后加入 5μL荧光探针(15nM)和5μL的UGM(KpUGM为500nm、MtUGM为580nM),酶标仪测定,激发光:485nm;发射光:535nm。每个样品平行试验3次。GraphPad Prism软件计算Kd值,结果见表3。
表3 UGM抑制活性结果
2.5牛型结核分枝杆菌的体外筛选试验
2.5.1在对上述32个化合物针对牛型结核分枝杆菌的体外筛选试验,当化合物浓度在500μg/mL时,有27例发明实施例对Mycobacterium Bovis(BCG)菌株有抑制作用。当化合物浓度在100μg/mL时,有12例发明实施例对Mycobacterium Bovis(BCG) 菌株有抑制作用。此外,发明人针对在上轮实验中抗结核活性较好的实施例设计了其对小鼠巨噬细胞RAW264.7的体外细胞毒性作用,并计算出选择性指数(SI= IC50/MIC)。SI比率越高意味着该化合物仅在高浓度下才具有细胞毒性,而在较低浓度下显示出抗结核活性。化合物SI大于1.0为有效,SI值越大则该化合物越安全。所有发明实施例对体外抗BCG活性测试结果,见表4:
表4体外抗BCG活性初步筛选结果
1.“-”表示该样品在该测试浓度下对牛型结核分枝杆菌菌株无效。
2.“+”表示该样品在该测试浓度下对牛型结核分枝杆菌菌株有效。
2.5.2体外抗BCG活性、小鼠巨噬细胞RAW264.7的半抑制浓度及选择性指数,见表5,图7、图8、图9、图10、图11、图12。
表5半抑制浓度及选择性指数结果
“-”表示该样品未测试
根据需要,可将本发明的化合物或其药用盐的药物,可以采用的剂型是惯用盖伦给药剂型,例如:膏药、片剂、丸剂、胶囊剂、栓剂、乳剂和注射剂。这些制剂按众所周知的方法,使用传统的添加剂和赋形剂制得。由此制得药物根据需要可以按局部、口服、注射等途径给药。
3结论
本发明通过生物电子等排策略,引入酰胺键代替迷迭香酸的酯键具有普适性,设计并制备了一系列结构修饰产物,结合酶水平和体外抗牛型结核分枝杆菌实验,表明本发明中涉及的化合物有较好的抗结核活性,为寻找具有良好开发前景的抗结核候选药物分子奠定基础。
虽然,上文中已经用一般性说明、具体实施方式及试验,对本发明作了详尽的描述,但在本发明基础上,可以对之作出一些修改或改进,这对本领域技术人员而言是显而易见的。因此,在不偏离本发明精神的基础上所做的这些修改或改进,均属于本发明要求保护的范围。
Claims (10)
3.根据权利要求1所述的迷迭香酸生物电子等排体,其特征在于,所述的迷迭香酸生物电子等排体包括单键和双键。
4.权利要求1或2所述迷迭香酸生物电子等排体的制备方法,其特征在于,所述制备方法具体步骤为:
a.称取2.0-3.0g,10mmoL,1eq的O-苄基-L-酪氨酸于反应瓶中,向瓶内注入50-80mL甲醇;将反应瓶置于冰盐浴下冷却10-30min;待体系温度降低至-10℃后,向反应瓶内缓慢滴加4.4mL,60mmoL,6eq重蒸的SOCl2,控制滴速,使体系温度保持在0℃以下;滴加完毕后,冰盐浴下反应5-15min,撤去冰盐浴后使反应瓶恢复至室温,再于60-100℃油浴下加热回流2-5h,TLC跟踪检测,反应结束后,减压回收溶剂,过滤,滤饼用二氯甲烷洗涤以获得所需化合物,将化合物真空抽干数小时后即得白色粉末状1;
b.称取5.0-6.0g,30mmoL,1eq的2,3-二氯苯甲醛于圆底烧瓶中,加入20-50mL吡啶并于80-100℃油浴中搅拌溶解样品;溶解后向反应瓶中加入9.0-10.0g,90mmoL,3eq的丙二酸并滴加880μL,9mmoL,0.3eq的哌啶于80-100℃油浴中加热回流2-5h后,TLC检测反应结束;反应体系冷却至室温后,将其倾入200-500mL冰水溶液中,用5-15%的盐酸调节pH值至4,有大量的白色固体析出;过滤,白色固体用蒸馏水洗2-3次;再用适量乙酸乙酯溶解白色固体,滤去不溶物;减压回收部分溶剂后置于冰箱中析出白色粉末状固体2;
c.称取0.4-0.5g,2mmoL,1eq的白色粉末状固体2置于反应瓶内,溶解于10-30mL无水DMF中,再加入0.5-0.6g,2mmoL,1eq的HBTU和695μL,4mmoL,2eq的DIEA,室温下反应20-50min;随后向反应物中加入0.5-0.6g,2mmoL,1eq的1,并于室温下反应10-15h;TLC检测反应结束后,加入50-100mL乙酸乙酯和20-50mL蒸馏水分散,萃取,依次以蒸馏水和饱和氯化钠溶液洗涤,无水MgSO4干燥,减压回收乙酸乙酯至干所得粗品经比例为3:1的石油醚:乙酸乙酯硅胶柱层析纯化,得白色固体3;
d.取白色固体3,加入5-10mL蒸馏水和30-40mg,1.5mmoL,1.5eq的LiOH,室温搅拌反应1-3h,TLC检测反应结束;用1M浓盐酸调pH至2-3;减压回收溶剂至干,所得残留物再经二氯甲烷洗涤后过滤,抽干滤饼得化合物4a,(N)-2-[(5,6-二氯基)-苯丙烯酰胺基]-3-(4-苄氧基苯基)-丙酸。
5.根据权利要求4所述的制备方法,其特征在于,所述制备方法具体步骤为:
a.称取2.71g,10mmoL,1eq的O-苄基-L-酪氨酸于反应瓶中,向瓶内注入60mL甲醇;将反应瓶置于冰盐浴下冷却20min;待体系温度降低至-10℃后,向反应瓶内缓慢滴加4.4mL,60mmoL,6eq重蒸的SOCl2,控制滴速,使体系温度保持在0℃以下;滴加完毕后,冰盐浴下反应10min,撤去冰盐浴后使反应瓶恢复至室温,再于80℃油浴下加热回流3h,TLC跟踪检测,反应结束后,减压回收溶剂,过滤,滤饼用二氯甲烷洗涤以获得所需化合物,将化合物真空抽干数小时后即得白色粉末状1;
b.称取5.26g,30mmoL,1eq的2,3-二氯苯甲醛于圆底烧瓶中,加入30mL吡啶并于100℃油浴中搅拌溶解样品;溶解后向反应瓶中加入9.36g,90mmoL,3eq的丙二酸并滴加880μL,9mmoL,0.3eq的哌啶于100℃油浴中加热回流3h后,TLC检测反应结束;反应体系冷却至室温后,将其倾入300mL冰水溶液中,用10%的盐酸调节pH值至4,有大量的白色固体析出;过滤,白色固体用蒸馏水洗2-3次;再用适量乙酸乙酯溶解白色固体,滤去不溶物;减压回收部分溶剂后置于冰箱中析出白色粉末状固体2;
c.称取0.44g,2mmoL,1eq的白色粉末状固体2置于反应瓶内,溶解于20mL无水DMF中,再加入0.58g,2mmoL,1eq的HBTU和695μL,4mmoL,2eq的DIEA,室温下反应30min;随后向反应物中加入0.57g,2mmoL,1eq的1,并于室温下反应14h;TLC检测反应结束后,加入80mL乙酸乙酯和40mL蒸馏水分散,萃取,依次以蒸馏水和饱和氯化钠溶液洗涤,无水MgSO4干燥,减压回收乙酸乙酯至干所得粗品经比例为3:1的石油醚:乙酸乙酯硅胶柱层析纯化,得白色固体3;
d.取白色固体3,加入5mL蒸馏水和36mg,1.5mmoL,1.5eq的LiOH,室温搅拌反应2h,TLC检测反应结束;用1M浓盐酸调pH至2-3;减压回收溶剂至干,所得残留物再经二氯甲烷洗涤后过滤,抽干滤饼得化合物4a,(N)-2-[(5,6-二氯基)-苯丙烯酰胺基]-3-(4-苄氧基苯基)-丙酸。
6.根据权利要求4或5所述的制备方法,其特征在于,所述制备方法可用于权利要求2所有衍生物的制备。
7.根据权利要求2所述的迷迭香酸生物电子等排体,其特征在于,所述的迷迭香酸生物电子等排体衍生物加入药学上可接受的辅料制成药学上可接受的制剂;所述制剂为片剂、丸剂、胶囊剂、栓剂、乳剂和注射剂。
8.根据权利要求7所述的迷迭香酸生物电子等排体,其特征在于,所述片剂制备方法为:取权利要求2任一衍生物的产物5g、羟丙甲纤维素6g、羧甲淀粉钠10g、微晶纤维素8g、乳糖115g、淀粉50g、硬脂酸镁1g;将主药与辅料充分混匀后投入高速搅拌机中,喷雾加水适量,整粒,水分控制在3~4%,然后压片,制成1000片,包薄膜衣。
9.如权利要求1或2所述的迷迭香酸生物电子等排体在制备抑制结核分枝杆菌细胞壁生物合成关键酶——UGM活性及在制备抗牛型结核分枝杆菌药物中的应用。
10.根据权利要求9所述的应用,其特征在于,对BCG菌株抑制浓度为5μg/mL-500μg/mL。
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN202211159907.3A CN115385819B (zh) | 2022-09-22 | 2022-09-22 | 一种迷迭香酸生物电子等排体及其制备方法和应用 |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN202211159907.3A CN115385819B (zh) | 2022-09-22 | 2022-09-22 | 一种迷迭香酸生物电子等排体及其制备方法和应用 |
Publications (2)
Publication Number | Publication Date |
---|---|
CN115385819A true CN115385819A (zh) | 2022-11-25 |
CN115385819B CN115385819B (zh) | 2024-05-24 |
Family
ID=84126008
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN202211159907.3A Active CN115385819B (zh) | 2022-09-22 | 2022-09-22 | 一种迷迭香酸生物电子等排体及其制备方法和应用 |
Country Status (1)
Country | Link |
---|---|
CN (1) | CN115385819B (zh) |
Cited By (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN117488458A (zh) * | 2023-10-25 | 2024-02-02 | 普宁市新栩纺织有限公司 | 一种抑菌透气面料 |
Citations (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN102731296A (zh) * | 2012-06-06 | 2012-10-17 | 中国人民解放军第二军医大学 | 迷迭香酸衍生物及其制备方法与在制备抗结核药物中的应用 |
-
2022
- 2022-09-22 CN CN202211159907.3A patent/CN115385819B/zh active Active
Patent Citations (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN102731296A (zh) * | 2012-06-06 | 2012-10-17 | 中国人民解放军第二军医大学 | 迷迭香酸衍生物及其制备方法与在制备抗结核药物中的应用 |
Non-Patent Citations (2)
Title |
---|
FU, JIAN等: "Synthesis and evaluation of inhibitors of Mycobacterium tuberculosis UGM using bioisosteric replacement", BIOORGANIC & MEDICINAL CHEMISTRY, vol. 69, pages 1 - 7 * |
夏于芬等: "迷迭香酸衍生物的合成", 中国化学会第八届有机化学学术会议暨首届重庆有机化学国际研讨会论文摘要集, pages 756 * |
Cited By (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN117488458A (zh) * | 2023-10-25 | 2024-02-02 | 普宁市新栩纺织有限公司 | 一种抑菌透气面料 |
Also Published As
Publication number | Publication date |
---|---|
CN115385819B (zh) | 2024-05-24 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
RU2183632C2 (ru) | Антагонисты эндотелина: n-[[2'-[[(4,5-диметил-3-изоксазолил)амино]сульфонил]-4-(2-оксазолил)[1,1'- бифенил]-2-ил]метил]-n,3,3-триметилбутанамид и n-(4,5-диметил-3-изоксазолил)-2'-[(3,3-диметил-2-оксо-1-пирролидинил)метил ]-4'-(2-оксазолил)[1,1'-бифенил]-2-сульфонамид и их соли | |
EA009196B1 (ru) | Ацетил 2-гидрокси-1,3-диаминоалканы | |
AU2020340299B2 (en) | Crystalline forms of a CD73 inhibitor | |
CN107445896B (zh) | 一种具有抗肿瘤活性的苯基异羟肟酸类化合物及其应用 | |
MXPA06003249A (es) | Acidos de heteroarilo benzofurano substituidos. | |
EP3611170B1 (en) | Deuterated compounds and medical use thereof as antianxiety agents | |
CN110627774A (zh) | 一种抗肝纤维化的化合物、制剂、制备方法和用途 | |
CN115385819B (zh) | 一种迷迭香酸生物电子等排体及其制备方法和应用 | |
JP6929866B2 (ja) | 2,3−エポキシスクシニル誘導体、その調製方法及び使用 | |
CN111454229B (zh) | 二氢萘并异噁唑类衍生物及其在抗肿瘤药物中的应用 | |
CN113444069A (zh) | 一类2-芳基-4-(1h-吡唑-3-基)吡啶类lsd1/hdac双靶点抑制剂 | |
CN109096272B (zh) | 一种具有抗肿瘤活性的吲哚异羟肟酸类化合物及其应用 | |
CN108752412B (zh) | 乳香酸衍生物及其应用 | |
CN115010658B (zh) | 一种化合物及其制备方法与应用 | |
CN110317137B (zh) | 9,10-蒽醌类化合物或其药学上可接受的盐及其药物用途 | |
FR2758329A1 (fr) | Derives d'imidazole-4-butane boronique, leur preparation et leur utilisation en therapeutique | |
TW202016071A (zh) | 四氫吖唉衍生物及其前藥 | |
CN113461563B (zh) | Nqo1激活型6-重氮基-5-氧代-l-正亮氨酸前药及其制备方法和应用 | |
RU2712140C2 (ru) | Композиции для лечения фиброза и связанных с фиброзом состояний | |
TW200521126A (en) | Chroman carboxylic acid derivatives for the treatment of diabetes and lipid disorders | |
CN110922450B (zh) | Psma激活式抗肿瘤前药cpt-x及其制备方法和应用 | |
CN111825608A (zh) | 四氢喹啉类与四氢异喹啉类化合物及其用途 | |
CN111333636B (zh) | 噻唑酰胺类衍生物及其在抗肿瘤药物中的应用 | |
WO2023169480A1 (zh) | 氘代化合物,及其制备方法和应用 | |
CN114702444B (zh) | 一种具有抗多发性骨髓瘤作用的aha1抑制剂及其制备方法和应用 |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
PB01 | Publication | ||
PB01 | Publication | ||
SE01 | Entry into force of request for substantive examination | ||
SE01 | Entry into force of request for substantive examination | ||
GR01 | Patent grant | ||
GR01 | Patent grant |