WO2018059537A1 - 一种二氮杂螺[5.5]十一碳烷衍生物及其用途 - Google Patents
一种二氮杂螺[5.5]十一碳烷衍生物及其用途 Download PDFInfo
- Publication number
- WO2018059537A1 WO2018059537A1 PCT/CN2017/104286 CN2017104286W WO2018059537A1 WO 2018059537 A1 WO2018059537 A1 WO 2018059537A1 CN 2017104286 W CN2017104286 W CN 2017104286W WO 2018059537 A1 WO2018059537 A1 WO 2018059537A1
- Authority
- WO
- WIPO (PCT)
- Prior art keywords
- ring
- ethyl
- group
- mmol
- diazaspiro
- Prior art date
Links
- 0 *c(cc(cc1N2)O)c1OCC2=O Chemical compound *c(cc(cc1N2)O)c1OCC2=O 0.000 description 10
- KDLMHKMMEUATCA-UHFFFAOYSA-N CC(C)(C)OC(N(CC1)CCC1(C1)OCCN1c1cc(Cl)ncc1)=O Chemical compound CC(C)(C)OC(N(CC1)CCC1(C1)OCCN1c1cc(Cl)ncc1)=O KDLMHKMMEUATCA-UHFFFAOYSA-N 0.000 description 1
- CMADDVGEXUPYGO-UHFFFAOYSA-N CC(C)(C)OC(N(CC1)CCC1(C1)OCCN1c1ccccc1)=O Chemical compound CC(C)(C)OC(N(CC1)CCC1(C1)OCCN1c1ccccc1)=O CMADDVGEXUPYGO-UHFFFAOYSA-N 0.000 description 1
- SMLRIWGQYOVYSP-UHFFFAOYSA-N CC(C)c1cc(N2CC3(CCN(Cc4cccc(CCOCCC(N(C)CCNCCc(c(OC5)c6NC5=O)ccc6O)=O)c4)CC3)OCC2)ncc1 Chemical compound CC(C)c1cc(N2CC3(CCN(Cc4cccc(CCOCCC(N(C)CCNCCc(c(OC5)c6NC5=O)ccc6O)=O)c4)CC3)OCC2)ncc1 SMLRIWGQYOVYSP-UHFFFAOYSA-N 0.000 description 1
- MCTPFWWSYVMUIA-QNGWXLTQSA-N CC(C)c1ccnc(N2CC3(CCN(Cc4cccc(CCOCCC(N(C)CCNC[C@@H](c(c(C=C5)c6NC5=O)ccc6O)O)=O)c4)CC3)OCC2)c1 Chemical compound CC(C)c1ccnc(N2CC3(CCN(Cc4cccc(CCOCCC(N(C)CCNC[C@@H](c(c(C=C5)c6NC5=O)ccc6O)O)=O)c4)CC3)OCC2)c1 MCTPFWWSYVMUIA-QNGWXLTQSA-N 0.000 description 1
- LUUKWTNSHASSOL-UHFFFAOYSA-N CC1(C)Oc2c(C)ccc(O)c2NC1=O Chemical compound CC1(C)Oc2c(C)ccc(O)c2NC1=O LUUKWTNSHASSOL-UHFFFAOYSA-N 0.000 description 1
- AAGHVHKJRZKNND-UHFFFAOYSA-N CCC(C)c(ccc(O)c1N2)c1OCC2=C Chemical compound CCC(C)c(ccc(O)c1N2)c1OCC2=C AAGHVHKJRZKNND-UHFFFAOYSA-N 0.000 description 1
- LMMHOPSKYOBKJH-UHFFFAOYSA-N CCCN(CCNCCc(c(OC1)c2NC1=O)ccc2O)C(CCOCCc1cccc(CN(CC2)CCC2(C2)OCCN2C(c2n[n](CC3CC3)cc2)=O)c1)=O Chemical compound CCCN(CCNCCc(c(OC1)c2NC1=O)ccc2O)C(CCOCCc1cccc(CN(CC2)CCC2(C2)OCCN2C(c2n[n](CC3CC3)cc2)=O)c1)=O LMMHOPSKYOBKJH-UHFFFAOYSA-N 0.000 description 1
- PGDZGMXAHNHMLW-UHFFFAOYSA-N CCc1cc(N2CC3(CCN(Cc(cccc4CCO)c4F)CC3)OCC2)ncc1 Chemical compound CCc1cc(N2CC3(CCN(Cc(cccc4CCO)c4F)CC3)OCC2)ncc1 PGDZGMXAHNHMLW-UHFFFAOYSA-N 0.000 description 1
- CQHKQLLXRYCWDZ-UHFFFAOYSA-N CCc1cc(N2CC3(CCN(Cc(cccc4CCOCCC(OC(C)(C)C)=O)c4F)CC3)OCC2)ncc1 Chemical compound CCc1cc(N2CC3(CCN(Cc(cccc4CCOCCC(OC(C)(C)C)=O)c4F)CC3)OCC2)ncc1 CQHKQLLXRYCWDZ-UHFFFAOYSA-N 0.000 description 1
- KKYSJTHHMIANMV-UHFFFAOYSA-N CCc1cc(N2CC3(CCN(Cc4cc(CCOCCC(N(C)CC(OC)OC)=O)ccc4)CC3)OCC2)ncc1 Chemical compound CCc1cc(N2CC3(CCN(Cc4cc(CCOCCC(N(C)CC(OC)OC)=O)ccc4)CC3)OCC2)ncc1 KKYSJTHHMIANMV-UHFFFAOYSA-N 0.000 description 1
- SAGVEVOUCGGTBU-UMSFTDKQSA-N CCc1ccnc(N2CC3(CCN(Cc4cc(CCOCCC(N(C)CCNC[C@@H](c(ccc(O)c5N6)c5OCC6=O)O)=O)ccc4)CC3)OCC2)c1 Chemical compound CCc1ccnc(N2CC3(CCN(Cc4cc(CCOCCC(N(C)CCNC[C@@H](c(ccc(O)c5N6)c5OCC6=O)O)=O)ccc4)CC3)OCC2)c1 SAGVEVOUCGGTBU-UMSFTDKQSA-N 0.000 description 1
- PNPBWLRUDBRJGD-BHVANESWSA-N CN(CCNC[C@@H](c(c(C=C1)c2NC1=O)ccc2O)O)C(CCOCCc1cc(CN(CC2)CCC2(C2)OCCN2c2nc(-c3ccc[s]3)ccn2)ccc1)=O Chemical compound CN(CCNC[C@@H](c(c(C=C1)c2NC1=O)ccc2O)O)C(CCOCCc1cc(CN(CC2)CCC2(C2)OCCN2c2nc(-c3ccc[s]3)ccn2)ccc1)=O PNPBWLRUDBRJGD-BHVANESWSA-N 0.000 description 1
- NVXWXNHWRNMNQX-UMSFTDKQSA-N CN(CCNC[C@@H](c(cc1)c(C=CC(N2)=O)c2c1O)O)C(CCOCCc1cccc(CN(CC2)CCC2(C2)OCCN2c2ccnc(Cl)c2)c1)=O Chemical compound CN(CCNC[C@@H](c(cc1)c(C=CC(N2)=O)c2c1O)O)C(CCOCCc1cccc(CN(CC2)CCC2(C2)OCCN2c2ccnc(Cl)c2)c1)=O NVXWXNHWRNMNQX-UMSFTDKQSA-N 0.000 description 1
- MMEXLVRGXKARPC-XIFFEERXSA-N CN(CCNC[C@@H](c1cc(O)cc(N2)c1OCC2=O)O)C(CCOCCc1cc(CN(CC2)CCC2(C2)OCCN2c2cc(F)ccn2)ccc1)=O Chemical compound CN(CCNC[C@@H](c1cc(O)cc(N2)c1OCC2=O)O)C(CCOCCc1cc(CN(CC2)CCC2(C2)OCCN2c2cc(F)ccn2)ccc1)=O MMEXLVRGXKARPC-XIFFEERXSA-N 0.000 description 1
- PGCFPSPVVBKJFB-UHFFFAOYSA-N COc(c(C[O](C)=C)c1)ccc1I Chemical compound COc(c(C[O](C)=C)c1)ccc1I PGCFPSPVVBKJFB-UHFFFAOYSA-N 0.000 description 1
- ABDKAPXRBAPSQN-UHFFFAOYSA-N COc1ccccc1OC Chemical compound COc1ccccc1OC ABDKAPXRBAPSQN-UHFFFAOYSA-N 0.000 description 1
- YOAJDUWSPDQYRK-UHFFFAOYSA-N C[O](c1cc([O](C)=C)ccc1)=C Chemical compound C[O](c1cc([O](C)=C)ccc1)=C YOAJDUWSPDQYRK-UHFFFAOYSA-N 0.000 description 1
- KMOFQISPDGIOIG-UHFFFAOYSA-N Cc(cc(cc1N2)OC)c1OC2=O Chemical compound Cc(cc(cc1N2)OC)c1OC2=O KMOFQISPDGIOIG-UHFFFAOYSA-N 0.000 description 1
- SBQUNTRLQMQGSF-UHFFFAOYSA-N Cc(cc1)c(C=CC(N2)=O)c2c1O Chemical compound Cc(cc1)c(C=CC(N2)=O)c2c1O SBQUNTRLQMQGSF-UHFFFAOYSA-N 0.000 description 1
- ZUVDVLYXIZFDRM-UHFFFAOYSA-N Cc(cc1)cc(CO)c1O Chemical compound Cc(cc1)cc(CO)c1O ZUVDVLYXIZFDRM-UHFFFAOYSA-N 0.000 description 1
- OVHWFHUMTTZMQW-UHFFFAOYSA-N Cc(cc1)cc(NS(C)(=O)=O)c1O Chemical compound Cc(cc1)cc(NS(C)(=O)=O)c1O OVHWFHUMTTZMQW-UHFFFAOYSA-N 0.000 description 1
- ZBCATMYQYDCTIZ-UHFFFAOYSA-N Cc(cc1)cc(O)c1O Chemical compound Cc(cc1)cc(O)c1O ZBCATMYQYDCTIZ-UHFFFAOYSA-N 0.000 description 1
- NQYGGOUCIQZRGU-UHFFFAOYSA-N Cc(ccc(O)c1N2)c1OCC2=O Chemical compound Cc(ccc(O)c1N2)c1OCC2=O NQYGGOUCIQZRGU-UHFFFAOYSA-N 0.000 description 1
- HPQMGHBJYJSCNS-UHFFFAOYSA-N Cc1c(CC(C=C)C(S2)=C)c2c(C)cc1 Chemical compound Cc1c(CC(C=C)C(S2)=C)c2c(C)cc1 HPQMGHBJYJSCNS-UHFFFAOYSA-N 0.000 description 1
- SJRRNEXDVMQLFZ-UHFFFAOYSA-N Cc1nc(N2CC3(CCN(Cc4cccc(CCOCCC(N(C)CCNCC(c(c(C=C5)c6NC5=O)ccc6O)O)=O)c4)CC3)OCC2)ncc1 Chemical compound Cc1nc(N2CC3(CCN(Cc4cccc(CCOCCC(N(C)CCNCC(c(c(C=C5)c6NC5=O)ccc6O)O)=O)c4)CC3)OCC2)ncc1 SJRRNEXDVMQLFZ-UHFFFAOYSA-N 0.000 description 1
- ONHMWUXYIFULDO-UHFFFAOYSA-N Clc1nccc(Br)c1 Chemical compound Clc1nccc(Br)c1 ONHMWUXYIFULDO-UHFFFAOYSA-N 0.000 description 1
- ZJRBRKUGRKKZOO-UHFFFAOYSA-N OCc(nccc1)c1O Chemical compound OCc(nccc1)c1O ZJRBRKUGRKKZOO-UHFFFAOYSA-N 0.000 description 1
- NXOVQYQJPVOXBC-UHFFFAOYSA-N OCc1cc(I)ccc1O Chemical compound OCc1cc(I)ccc1O NXOVQYQJPVOXBC-UHFFFAOYSA-N 0.000 description 1
- WYWNNDBGZYGIHZ-QNGWXLTQSA-N O[C@@H](CNCCN(C1CC1)C(CCOCCc1cc(CN(CC2)CCC2(C2)OCCN2C(c2n[n](CC3CC3)cc2)=O)ccc1)=O)c(c(OC1)c2NC1=O)ccc2O Chemical compound O[C@@H](CNCCN(C1CC1)C(CCOCCc1cc(CN(CC2)CCC2(C2)OCCN2C(c2n[n](CC3CC3)cc2)=O)ccc1)=O)c(c(OC1)c2NC1=O)ccc2O WYWNNDBGZYGIHZ-QNGWXLTQSA-N 0.000 description 1
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/33—Heterocyclic compounds
- A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
- A61K31/41—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having five-membered rings with two or more ring hetero atoms, at least one of which being nitrogen, e.g. tetrazole
- A61K31/425—Thiazoles
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/33—Heterocyclic compounds
- A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
- A61K31/535—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with at least one nitrogen and one oxygen as the ring hetero atoms, e.g. 1,2-oxazines
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P11/00—Drugs for disorders of the respiratory system
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P11/00—Drugs for disorders of the respiratory system
- A61P11/06—Antiasthmatics
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P11/00—Drugs for disorders of the respiratory system
- A61P11/08—Bronchodilators
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D498/00—Heterocyclic compounds containing in the condensed system at least one hetero ring having nitrogen and oxygen atoms as the only ring hetero atoms
- C07D498/02—Heterocyclic compounds containing in the condensed system at least one hetero ring having nitrogen and oxygen atoms as the only ring hetero atoms in which the condensed system contains two hetero rings
- C07D498/10—Spiro-condensed systems
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D519/00—Heterocyclic compounds containing more than one system of two or more relevant hetero rings condensed among themselves or condensed with a common carbocyclic ring system not provided for in groups C07D453/00 or C07D455/00
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
- Y02P20/00—Technologies relating to chemical industry
- Y02P20/50—Improvements relating to the production of bulk chemicals
- Y02P20/55—Design of synthesis routes, e.g. reducing the use of auxiliary or protecting groups
Definitions
- the present invention relates to a diazaspiro undecane derivatives, preparation methods and uses [5.5] in medicine, in particular a muscarinic receptor antagonist and ⁇ having 2 - adrenergic receptor agonistic Double-active novel diazaspiro[5.5]undecane or a stereoisomer, hydrate, solvate, metabolite, pharmaceutically acceptable salt, co-crystal or prodrug thereof, pharmaceutical composition thereof Its application in medicine.
- Bronchodilators play an important role in the treatment of respiratory diseases such as chronic obstructive pulmonary disease (COPD) and asthma.
- Bronchodilators in clinical use include widely muscarinic receptor antagonist and ⁇ 2 - adrenergic agonist.
- a muscarinic receptor antagonist exerts potency in bronchodilation by reducing the level of vagal cholinergic energy in airway smooth muscle.
- Inhaled muscarinic receptor antagonists currently used include ipratropium bromide, oxitropium bromide, glycopyrrolate, tiotropium bromide, adiponium bromide and indole bromide.
- 2 2 -adrenergic agonists bronchodilate by stimulating adrenergic receptors in airway smooth muscle, reversing the response of bronchoconstrictors to various mediators such as acetylcholine.
- the ⁇ 2 -adrenergic agonists currently used include albuterol, salmeterol, arformoterol, formoterol, vilantrol and indacaterol. In addition to improving lung function, these drugs can improve the quality of life of patients and reduce the deterioration of their condition.
- the combination of muscarinic receptor antagonists and ⁇ 2 -adrenergic agonists is more effective than the use of one of the therapeutic agents alone
- the current prion base receptor antagonists And ⁇ 2 -adrenergic agonists are prepared as a combination preparation for the treatment of asthma and moderate to severe COPD.
- These compound preparations mainly include Anoro Ellipta (Ambroxol/Vylantro) and Ultibro Breezhaler (Glon Brom). Ammonium / indacaterol) and ipratropium bromide / salbutamol.
- muscarinic receptor antagonist and ⁇ 2 - adrenergic agonists dual-acting drugs drugs with this dual function the advantages of both pharmaceutical ingredient combination, along with a single molecule pharmacokinetics.
- These compounds are administered as a single therapeutic agent and can provide bronchodilation by two different and possibly synergistic modes of action.
- compounds with muscarinic receptor antagonism and ⁇ 2 -adrenergic agonistic dual action can also be combined with corticosteroid (ICS) anti-inflammatory drugs to form two therapeutic agents (MABA/ICS) to provide triple Therapeutic effect of action (Expert Opin. Investig. Drugs (2014) 23(4): 453-456).
- ICS corticosteroid
- the present invention provides a compound of the formula (I) or a stereoisomer, hydrate, metabolite, solvate, pharmaceutically acceptable salt, co-crystal or prodrug thereof:
- R 1 is selected from a C 6-12 carbocyclic ring or a 5- to 12-membered heterocyclic ring, and the carbocyclic or heterocyclic ring is optionally further substituted with 0 to 5 R 1a , and the heterocyclic ring contains 1 to 4 a hetero atom selected from N, O or S;
- R 1 is substituted with from 1 to 5 R 1a , and at least one R 1a is selected from C 2-6 alkynyl or -CH 2 -C 3-6 ring alkyl;
- R 2 , R 3 , R 4 , and R 5 are each independently selected from a C 1-6 alkylene group, and the alkylene group is further optionally further selected from 0 to 4 selected from the group consisting of F, Cl, Br, I, and cyano. Substituted with a substituent of a hydroxy group, a C 1-6 alkyl group, a C 1-6 alkoxy group, a phenyl group or a phenyl-C 1-4 alkylene group;
- A is selected from a C 3-12 carbocyclic ring or a 5 to 12 membered heterocyclic ring, and the carbocyclic or heterocyclic ring is optionally further further 0 to 5 F, Cl, Br, I, cyano, hydroxy, amino, C 1 Substituted with a substituent of -6 alkyl, C 2-6 alkynyl, C 1-4 alkoxy or C 3-6 cycloalkyl, and said heterocyclic ring contains 1 to 3 selected from N, O or S Hetero atom
- R x is each independently selected from H, C 1-6 alkyl or C 3-8 cycloalkyl, and the alkyl or cycloalkyl group is further further selected from 0 to 5 selected from F, Cl, Br, I, Substituted with a substituent of a C 1-4 alkyl group, a C 3-6 cycloalkyl group or a C 1-4 alkoxy group;
- R 3 -X 1 -R 4 -X 2 -R 5 may be an ethylene group
- R 6 and R 7 are each independently selected from H or C 1-4 alkyl
- R 8 is selected from H or OH
- a preferred embodiment of the invention a compound of the formula (I) or a stereoisomer, hydrate, metabolite, solvate, pharmaceutically acceptable salt, co-crystal or prodrug thereof, wherein:
- a preferred embodiment of the invention a compound of the formula (I) or a stereoisomer, hydrate, metabolite, solvate, pharmaceutically acceptable salt, co-crystal or prodrug thereof, wherein:
- R 1 is selected from a C 6-10 carbocyclic ring or a 5- to 10-membered heterocyclic ring, preferably a benzene ring, a thiophene ring, a furan ring, a pyrrole ring, a thiazole ring, an oxazole ring, an imidazole ring, an isothiazole ring, an isoxazole ring.
- pyrazole ring pyridine ring, quinoline ring, isoquinoline ring, benzisothiazole ring, indazole ring, 1H-pyrazole [4,3-b]pyridine ring or
- R 1 is substituted with from 1 to 5 R 1a , and at least one R 1a is selected from C 2-4 alkynyl or -CH 2 -C 3-6 ring alkyl;
- R 2 and R 5 are each independently selected from a C 1-4 alkylene group, and the alkylene group is optionally further further selected from 0 to 4 selected from the group consisting of F, methyl, ethyl, methoxy or ethoxy. Substituted by a substituent;
- R 3 and R 4 are each independently selected from C 1-4 alkylene, preferably methylene, ethylene, propylene, -CH 2 CH(CH 3 )-, -CH(CH 3 )CH 2 - , -C(CH 3 ) 2 CH 2 -, -CH 2 C(CH 3 ) 2 -, butylene, -CH(CH 3 )CH 2 CH 2 -, -CH 2 CH(CH 3 )CH 2 - or -CH 2 CH(CH 3 )CH 2 -, the alkylene, methylene, ethylene, propylene or butylene group optionally further from 0 to 4 selected from F, methyl, ethyl , propyl, isopropyl, butyl, methoxy, ethoxy, phenyl, phenyl-methylene, phenyl-ethylene, phenyl-propylene or phenyl-butylene Substituted by
- A is selected from a C 6-10 carbocyclic ring or a 5 to 10 membered heterocyclic ring, preferably The carbon ring, heterocyclic ring, Optionally further by 0 to 5 F, Cl, Br, I, cyano, hydroxy, amino, C 1-6 alkyl, C 2-6 alkynyl, C 1-4 alkoxy or C 3-6 ring Substituted with a substituent of an alkyl group, and the heterocyclic ring contains 1 to 3 hetero atoms selected from N, O or S;
- R x is each independently selected from H, C 1-6 alkyl or C 3-8 cycloalkyl, and the alkyl or cycloalkyl group is further further selected from 0 to 5 selected from F, Cl, Br, I, Substituted with a substituent of methyl, ethyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, methoxy or ethoxy;
- R x is preferably H, methyl, ethyl, propyl, isopropyl, butyl, isobutyl, pentyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, -CH 2 -cyclopropyl , -CH 2 -cyclobutyl or -CH 2 -cyclopentyl;
- R 3 -X 1 -R 4 -X 2 -R 5 may be an ethylene group
- R 6 and R 7 are each independently selected from H or C 1-4 alkyl, preferably H, methyl or ethyl;
- R 8 is selected from H or OH
- a preferred embodiment of the invention a compound of the formula (I) or a stereoisomer, hydrate, metabolite, solvate, pharmaceutically acceptable salt, co-crystal or prodrug thereof, wherein:
- R 1 is selected from the group consisting of a benzene ring, a thiophene ring, a furan ring, a pyrrole ring, a thiazole ring, an oxazole ring, an imidazole ring, an isothiazole ring, an isoxazole ring, a pyrazole ring, a pyridine ring, a pyrimidine ring, a pyrazine ring, and a quinazine.
- a porphyrin ring an isoquinoline ring, a benzisothiazole ring, an indazole ring, a 1H-pyrazole [4,3-b]pyridine ring or
- R 1 is substituted with from 1 to 5 R 1a , and at least one R 1a is selected from ethynyl, propynyl, propargyl or -CH 2 - rings Propyl;
- R 2 and R 5 are each independently selected from a C 1-4 alkylene group, and the alkylene group is optionally further further selected from 0 to 4 selected from the group consisting of F, methyl, ethyl, methoxy or ethoxy. Substituted by a substituent;
- R 2 and R 5 are preferably methylene, ethylene, propylene, -CH 2 CH(CH 3 )-, -CH(CH 3 )CH 2 -, -C(CH 3 ) 2 CH 2 -, - CH 2 C(CH 3 ) 2 -, butylene, -CH(CH 3 )CH 2 CH 2 -, -CH 2 CH(CH 3 )CH 2 - or -CH 2 CH(CH 3 )CH 2 -;
- R 3 and R 4 are each independently selected from a methylene group, an ethylene group, a propylene group or a butylene group, and the methylene group, ethylene group, propylene group or butylene group is further optionally from 0 to 4 Selected from F, methyl, ethyl, propyl, isopropyl, butyl, methoxy, ethoxy, phenyl, phenyl-methylene, phenyl-ethylene, phenyl-propylene Substituted with a substituent of a phenyl-butylene group;
- R x is each independently selected from the group consisting of H, methyl, ethyl, propyl, isopropyl, butyl, isobutyl, pentyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, -CH 2 - Cyclopropyl, -CH 2 -cyclobutyl or -CH 2 -cyclopentyl;
- R 3 -X 1 -R 4 -X 2 -R 5 may be an ethylene group
- R 6 and R 7 are each independently selected from H, methyl or ethyl
- a preferred embodiment of the invention a compound of the formula (II) or formula (III) or a stereoisomer, hydrate, metabolite thereof, solvation thereof a pharmaceutically acceptable salt, eutectic or prodrug, wherein:
- P1 is selected from 0 or 1;
- P2 is selected from 0, 1, 2, 3 or 4;
- R a is selected from H, F, Cl, Br, I, C 1-4 alkyl, C 1-4 alkoxy or C 3-6 cycloalkyl;
- R 1 is selected from a C 6-10 carbocyclic ring or a 5 to 10 membered heterocyclic ring, and the carbocyclic or heterocyclic ring is optionally further substituted with 0 to 5 R 1a , and the heterocyclic ring contains 1 to 4 a hetero atom selected from N, O or S;
- R 1 is substituted with from 1 to 5 R 1a , and at least one R 1a is selected from C 2-4 alkynyl or -CH 2 -C 3-6 ring alkyl;
- R x is each independently selected from H, C 1-6 alkyl or C 3-8 cycloalkyl, and the alkyl or cycloalkyl group is further further selected from 0 to 5 selected from F, Cl, Br, I, Substituted with a substituent of methyl, ethyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, methoxy or ethoxy;
- R x is preferably H, methyl, ethyl, propyl, isopropyl, butyl, isobutyl, pentyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, -CH 2 -cyclopropyl , -CH 2 -cyclobutyl or -CH 2 -cyclopentyl;
- R 8 is selected from H or a hydroxyl group
- a preferred embodiment of the invention a compound of the formula (II) or formula (III) or a stereoisomer, hydrate, metabolite, solvate, pharmaceutically acceptable salt, eutectic or prodrug thereof, among them:
- P1 is selected from 0 or 1;
- P2 is selected from 0, 1, 2, 3 or 4;
- R 1 is selected from the group consisting of a benzene ring, a thiophene ring, a furan ring, a pyrrole ring, a thiazole ring, an oxazole ring, an imidazole ring, an isothiazole ring, an isoxazole ring, a pyrazole ring, a pyridine ring, a pyrazine ring, a pyrimidine ring, and a quinine.
- a porphyrin ring an isoquinoline ring, a benzisothiazole ring, an indazole ring, a 1H-pyrazole [4,3-b]pyridine ring or
- R 1 is substituted with from 1 to 5 R 1a , and at least one R 1a is selected from ethynyl, propynyl, propargyl, or -CH 2 - Cyclopropyl;
- R a is selected from the group consisting of F, Cl, Br, I, methyl, ethyl, methoxy, ethoxy or cyclopropane;
- R x is selected from the group consisting of H, methyl, ethyl, propyl, isopropyl, butyl, isobutyl, pentyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, -CH 2 -cyclopropane , -CH 2 -cyclobutyl or -CH 2 -cyclopentyl;
- a preferred embodiment of the invention a compound of the formula (II) or formula (III) or a stereoisomer, hydrate, metabolite, solvate, pharmaceutically acceptable salt, eutectic or prodrug thereof, among them:
- P1 is selected from 0 or 1;
- P2 is selected from 0, 1, 2, 3 or 4;
- R 1- W is selected from
- R a is selected from the group consisting of F, Cl, Br, I, methyl, ethyl, methoxy, ethoxy or cyclopropane;
- R x is selected from the group consisting of H, methyl, ethyl, propyl, isopropyl, butyl, isobutyl, pentyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, -CH 2 -cyclopropane , -CH 2 -cyclobutyl or -CH 2 -cyclopentyl;
- the compounds of the invention include, but are not limited to, one of the following compounds:
- the invention further relates to a pharmaceutical composition
- a pharmaceutical composition comprising a therapeutically effective amount of a compound of any one of formula (I), (II) or (III) or a stereoisomer thereof, hydrated , metabolite, solvate, pharmaceutically acceptable salt, eutectic or prodrug, and a pharmaceutically acceptable carrier, diluent, adjuvant, vehicle or excipient; said composition may be further One or more additional therapeutic agents are included; preferably, wherein the additional therapeutic agent is selected from one or more of a PDE4 inhibitor, a M receptor antagonist, a corticosteroid, and a beta-adrenoreceptor agonist.
- the invention further relates to providing a compound of the formula (I), (II) or (III) or a stereoisomer, hydrate, metabolite, solvate, pharmaceutically acceptable salt, eutectic or prodrug thereof Or the use of the aforementioned pharmaceutical composition for the preparation of a medicament for treating an airway obstructive disease, preferably, for the preparation of a medicament for the treatment of asthma, chronic obstructive pulmonary disease or bronchitis.
- the present invention also provides a method for treating an airway obstructive disease, the method comprising administering a compound of the formula (I), (II) or (III) or a stereoisomer, hydrate or metabolite thereof A solvate, a pharmaceutically acceptable salt, a eutectic or a prodrug, or a pharmaceutical composition as described above, preferably, the airway obstructive disease is selected from the group consisting of asthma, chronic obstructive pulmonary disease or bronchitis.
- the present invention also provides an intermediate for preparing a compound of the formula (I), which is selected from a compound represented by the formula (M-1), the formula (M-2) or the formula (M-3) or a stereospecific Constructs and pharmaceutically acceptable salts:
- R m1 is selected from an amino protecting group or H
- R m2 is selected from the group consisting of -COOH, -COOC 1-6 alkyl, -X 2 -R 5 -OH, -X 2 -R 5 - leaving group, -X 2 -R m3 CHO, -X 2 -R m3 C(OC 1-6 alkyl) 2 or
- the alkyl group is optionally further substituted with 0 to 4 F, Cl, Br or C 1-4 alkyl groups;
- R m3 is selected from an alkylene group having one carbon atom less than R 5 ;
- R 1 is selected from a C 6-12 carbocyclic ring or a 5- to 12-membered heterocyclic ring, preferably a C 6-10 carbocyclic ring or a 5- to 10-membered heterocyclic ring, more preferably a benzene ring, a thiophene ring, a furan ring, a pyrrole ring, a thiazole Ring, oxazole ring, imidazole ring, isothiazole ring, isoxazole ring, pyrazole ring, pyridine ring, pyrimidine ring, pyrazine ring, quinoline ring, isoquinoline ring, benzisothiazole ring, indazole ring , 1H-pyrazole [4,3-b]pyridine ring or The carbocyclic ring, heterocyclic ring, benzene ring, thiophene ring, furan ring, pyrrole ring,
- R 2 , R 3 , R 4 , and R 5 are each independently selected from a C 1-6 alkylene group, preferably a C 1-4 alkylene group, and the alkylene group is optionally further selected from 0 to 4 selected from F Substituted with a substituent of Cl, Br, I, cyano, hydroxy, C 1-6 alkyl, C 1-6 alkoxy, phenyl or phenyl-C 1-4 alkylene;
- R 2 and R 5 are methylene, ethylene, propylene, -CH 2 CH(CH 3 )-, -CH(CH 3 )CH 2 -, -C(CH 3 ) 2 CH 2 -, -CH 2 C(CH 3 ) 2 -, butylene, -CH(CH 3 )CH 2 CH 2 -, -CH 2 CH(CH 3 )CH 2 - or -CH 2 CH(CH 3 )CH 2 -;
- R 3 and R 4 are a methylene group, an ethylene group, a propylene group or a butylene group;
- A is selected from a C 3-12 carbocyclic ring or a 5 to 12 membered heterocyclic ring, preferably a C 6-10 carbocyclic ring or a 5 to 10 membered heterocyclic ring, and the carbocyclic or heterocyclic ring is optionally further further 0 to 5 F, Substituted with a substituent of Cl, Br, I, cyano, hydroxy, amino, C 1-6 alkyl, C 2-6 alkynyl, C 1-4 alkoxy or C 3-6 cycloalkyl, and said The heterocyclic ring contains 1 to 3 hetero atoms selected from N, O or S;
- R x is each independently selected from H, C 1-6 alkyl or C 3-8 cycloalkyl, and the alkyl or cycloalkyl group is further further selected from 0 to 5 selected from F, Cl, Br, I, Substituted with a substituent of a C 1-4 alkyl group, a C 3-6 cycloalkyl group or a C 1-4 alkoxy group;
- R x is preferably H, C 1-6 alkyl or C 3-8 cycloalkyl, said alkyl or cycloalkyl optionally further from 0 to 5 selected from the group consisting of F, Cl, Br, I, methyl, Substituted with a substituent of an ethyl group, a cyclopropyl group, a cyclobutyl group, a cyclopentyl group, a cyclohexyl group, a methoxy group or an ethoxy group;
- R x is more preferably H, methyl, ethyl, propyl, isopropyl, butyl, isobutyl, pentyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, -CH 2 -cyclopropane , -CH 2 -cyclobutyl or -CH 2 -cyclopentyl;
- the leaving group is selected from the group consisting of chlorine, bromine, iodine, -OMs or OTs;
- N1 is selected from 1, 2 or 3;
- R 1- W is selected from
- R m1 is selected from amino protecting group or H, preferably tert-butyloxycarbonyl or H;
- R m2 is selected from the group consisting of -COOH, -COOC 1-6 alkyl, -X 2 -R 5 -OH, -X 2 -R 5 - leaving group, -X 2 -R m3 CHO, -X 2 -R m3 C(OC 1-6 alkyl) 2 or
- the alkyl group is optionally further substituted by 0 to 4 F, Cl, Br or C 1-4 alkyl groups, and R m2 is preferably -COOH, -COO t- Bu or -X 2 -R m3 C(OCH 3 ) 2 ;
- R m3 is selected from an alkylene group having one carbon atom less than R 5 , and R m3 is preferably a methylene group or an ethylene group;
- X 1 is selected from -O-;
- R x is selected from the group consisting of H, methyl, ethyl, propyl, isopropyl, butyl, isobutyl, pentyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, -CH 2 -cyclopropane , -CH 2 -cyclobutyl or -CH 2 -cyclopentyl;
- R 2 , R 3 , R 4 , and R 5 are each independently selected from a methylene group, an ethylene group, or a propylene group;
- the leaving group is selected from the group consisting of chlorine, bromine, iodine, -OMs or OTs;
- N1 is selected from 1, 2 or 3.
- the preparation of the preferred compounds of the invention is optionally prepared by one of the following routes (the following reaction steps can be prepared by "one-pot”):
- R 1- W is selected from
- R x is selected from the group consisting of H, methyl, ethyl, propyl, isopropyl, butyl, isobutyl, pentyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, -CH 2 -cyclopropane , -CH 2 -cyclobutyl or -CH 2 -cyclopentyl;
- R a is selected from H, F, Cl, Br, methyl, ethyl, isopropyl or CF 3 ;
- the compound A (or a salt thereof, preferably a trifluoroacetate salt) is subjected to a substitution reaction with the compound B under basic conditions to obtain a compound C, preferably under the condition that the reaction substrate is dissolved in dioxane, acetonitrile, tetrahydrofuran, 2-methyl.
- a substitution reaction with the compound B under basic conditions to obtain a compound C, preferably under the condition that the reaction substrate is dissolved in dioxane, acetonitrile, tetrahydrofuran, 2-methyl.
- tetrahydrofuran 1 to 6 equivalents of sodium t-butoxide, potassium phosphate
- cesium carbonate or potassium carbonate a small amount of water (3% to 5% (v/v) of the reaction solvent) may be added, and the reaction is carried out at room temperature to reflux.
- Compound C is hydrolyzed to form compound D (or a salt thereof, preferably trifluoroacetate), preferably under conditions such that the reaction substrate is dissolved in dichloromethane and trifluoroacetic acid is added (1/5 to 1/min of dichloromethane). 1), reaction at room temperature.
- Compound E is substituted with compound D (or a salt thereof, preferably trifluoroacetate) to give compound F, preferably under conditions such that the reaction substrate is dissolved in dichloromethane in 1 to 6 equivalents of triethylamine, 4- In the presence of dimethylaminopyridine or N,N-diisopropylethylamine, 1.0 to 3 equivalents of HATU are added and reacted at room temperature.
- Compound F is reacted under deprotected conditions to give compound G.
- the reaction substrate is dissolved in tetrahydrofuran, and 1 to 10 equivalents of acetic acid or p-toluenesulfonic acid monohydrate are added, and the reaction is carried out at room temperature to 50 °C.
- Compound G is reductively aminated with compound H1 to obtain compound T1 (a preferred compound of the present invention), preferably under the condition that the reaction substrate is dissolved in N-methylpyrrolidone or N,N-dimethylformamide, and 0.8 to be added. After stirring 2 hours of acetic acid at room temperature for 0.5 to 1 hour, 2 to 3 equivalents of sodium triacetoxyborohydride are added and reacted at room temperature.
- Compound T is deprotected by TBS to give compound T2 (a preferred compound of the invention), preferably under conditions such that the reaction substrate is dissolved in tetrahydrofuran, and 5 to 20 equivalents of triethylamine trihydrofluoric acid are added and reacted at room temperature.
- Compound A (or a salt thereof, preferably a trifluoroacetate salt) is substituted with compound J under basic conditions to give compound K, preferably under conditions such that the reaction substrate is dissolved in dioxane, acetonitrile, tetrahydrofuran, 2-methyl In the tetrahydrofuran, a small amount of water (3% to 5% (v/v) of the reaction solvent) may be added in the presence of 1 to 6 equivalents of sodium t-butoxide, potassium phosphate, cesium carbonate or potassium carbonate, and room temperature ⁇ Reflow reaction.
- the compound A and the hydroxy-protected compound J are subjected to a substitution reaction under basic conditions, and then reacted under dehydroxylation conditions to obtain a compound K, wherein a preferred condition of the substitution reaction is to dissolve the reaction substrate in dioxane.
- a preferred condition of the substitution reaction is to dissolve the reaction substrate in dioxane.
- acetonitrile, tetrahydrofuran or 2-methyltetrahydrofuran a small amount of water (3% to 5 times of the reaction solvent) may be added in the presence of 1 to 6 equivalents of sodium t-butoxide, potassium phosphate, cesium carbonate or potassium carbonate. %(v/v)), room temperature to reflux reaction.
- the hydroxy protecting group is TBS
- the substrate is dissolved in tetrahydrofuran, and TBAF (tetrabutylammonium fluoride) is added and reacted at room temperature.
- Compound K is reacted with tert-butyl acrylate to form compound C, preferably under conditions such that the reaction substrate is dissolved in acetonitrile and benzyltrimethylammonium hydroxide is added and reacted at room temperature.
- Compound K can also be obtained by the following route:
- Compound A (or a salt thereof, preferably a trifluoroacetate salt) is reductively aminated with compound N to obtain compound K.
- the reaction substrate is dissolved in tetrahydrofuran, stirred at room temperature for 0.5 to 1 hour, and then added to 2 to 3 Equivalent sodium triacetoxyborohydride is reacted at room temperature.
- compound A undergoes a reductive amination reaction with a hydroxy-protected compound N and then reacts under deprotected conditions to form compound K.
- the hydroxy protecting group is TBS
- the substrate is dissolved in tetrahydrofuran, and TBAF (tetrabutylammonium fluoride) is added and reacted at room temperature.
- Compound C can also be obtained by the following route:
- Compound A (or a salt thereof, preferably a trifluoroacetate salt) is reductively aminated with compound M to obtain compound C.
- the reaction substrate is dissolved in tetrahydrofuran, stirred at room temperature for 0.5 to 1 hour, and then added to 2 to 3 Equivalent sodium triacetoxyborohydride is reacted at room temperature.
- Compound A (or a salt thereof, preferably trifluoroacetate) can be obtained by a route reaction:
- LG is selected from Cl, Br or I;
- R 1 is consistent with the above synthetic route
- PG is selected from the group consisting of tert-butyloxycarbonyl
- N-dimethylformamide 0.8 to 3 equivalents of 1,5-diazabicyclo[5.4.0]undec-5-ene, potassium carbonate, sodium carbonate, triethylamine, cesium carbonate or carbonic acid
- the reaction is carried out at room temperature to 150 ° C in the presence of potassium.
- the active agent preferably 1-propylphosphonic anhydride (T3P, CAS: 68957-94-8), HATU (CAS: 148893-10-1 or carbonyl diimidazole) is reacted at room temperature.
- the carbon, hydrogen, oxygen, sulfur, nitrogen or halogen involved in the groups and compounds of the present invention include their isotopes, and the carbon, hydrogen, oxygen, sulfur, and the groups and compounds involved in the present invention.
- the nitrogen or halogen is optionally further replaced by one or more of their corresponding isotopes, wherein the carbon isotopes include 12 C, 13 C, and 14 C, and the hydrogen isotopes include helium (H), helium (D, also known as heavy hydrogen ), ⁇ (T, also known as super heavy hydrogen), oxygen isotopes include 16 O, 17 O and 18 O, sulfur isotopes include 32 S, 33 S, 34 S and 36 S, nitrogen isotopes including 14 N and 15 N, the fluorine isotope 19 F, the chlorine isotope includes 35 Cl and 37 Cl, and the bromine isotopes include 79 Br and 81 Br.
- Alkyl means a straight-chain and branched monovalent saturated hydrocarbon group, and the main chain includes 1 to 10 carbon atoms, preferably 1 to 8 carbon atoms, further preferably 1 to 6 carbon atoms, more preferably 1 Straight and branched groups of up to 4 carbon atoms, most preferably 1 to 2 carbon atoms, examples of which include, but are not limited to, methyl, ethyl, n-propyl, isopropyl, n-butyl, iso Butyl, sec-butyl, tert-butyl, n-pentyl, 2-pentyl, 3-pentyl, 2-methyl-2-butyl, 3-methyl-2-butyl, n-hexyl, n-glycol a base, an n-octyl group, a n-decyl group, a n-decyl group, etc.; the alkyl group may optionally be further from 0 to 5 selected from the alky
- Alkoxy means a monovalent group of an O-alkyl group, wherein alkyl is as defined herein, and alkoxy embodiments include, but are not limited to, methoxy, ethoxy, 1-propoxy, 2 -propoxy, 1-butoxy, 2-methyl-1-propoxy, 2-butoxy, 2-methyl-2-propoxy, 1-pentyloxy, 2-pentyloxy , 3-pentyloxy, 2-methyl-2-butoxy, 3-methyl-2-butoxy, 3-methyl-1-butoxy and 2-methyl-1-butoxy Wait.
- alkenyl means a straight-chain or branched monovalent unsaturated hydrocarbon group having at least 1, usually 1, 2 or 3 carbon-carbon double bonds, the main chain comprising 2 to 10 carbon atoms, further preferably 2 Up to 6 carbon atoms, more preferably 2 to 4 carbon atoms in the main chain, and alkenyl examples include, but are not limited to, vinyl, allyl, 1-propenyl, 2-propenyl, 1-butenyl , 2-butenyl, 3-butenyl, 1-pentenyl, 2-pentenyl, 3-pentenyl, 4-pentenyl, 1-methyl-1-butenyl, 2- Methyl-1-butenyl, 2-methyl-3-butenyl, 1-hexenyl, 2-hexenyl, 3-hexenyl, 4-hexenyl, 5-hexenyl, 1-methyl-1-pentenyl, 2-methyl-1-pentenyl, 1-heptenyl, 2-heptenyl, 3-hep
- Alkynyl means a straight-chain or branched monovalent unsaturated hydrocarbon group having at least 1, usually 1, 2 or 3 carbon-carbon triple bonds, the main chain comprising 2 to 10 carbon atoms, further preferably 2 Up to 6 carbon atoms, more preferably 2 to 4 carbon atoms in the main chain
- alkynyl groups include, but are not limited to, ethynyl, 1-propynyl, 2-propynyl, butynyl, 2-butyl Alkynyl, 3-butynyl, 1-methyl-2-propynyl, 4-pentynyl, 3-pentynyl, 1-methyl-2-butynyl, 2-hexynyl, 3 -hexynyl, 2-heptynyl, 3-heptynyl, 4-heptynyl, 3-octynyl, 3-decynyl and 4-decynyl, etc.; the alkynyl group
- Cycloalkyl means a monovalent saturated carbocyclic hydrocarbon group, usually having from 3 to 10 carbon atoms, and non-limiting examples include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl or cycloheptyl.
- Carbocycle or “carbocyclyl” means a saturated or unsaturated aromatic or non-aromatic ring.
- the aromatic or non-aromatic ring may be a 3 to 10 membered monocyclic ring, a 4 to 12 membered bicyclic ring or a 10 to 15 membered member.
- a tricyclic system, a carbocyclic group may be bonded to a bridged ring or a spiro ring, and non-limiting examples include cyclopropyl, cyclobutyl, cyclopentyl, 1-cyclopentyl-1-enyl, 1-cyclopentyl -2-alkenyl, 1-cyclopentyl-3-alkenyl, cyclohexyl, 1-cyclohexyl-2-alkenyl, 1-cyclohexyl-3-alkenyl, cyclohexenyl, cyclohexadienyl , cycloheptyl, cyclooctyl, cyclodecyl, cyclodecyl, cyclodecyl, cyclododecyl, phenyl, naphthyl, benzocyclopropenyl, 2,3-dihydrobenzo ring Propylene based,
- the carbocyclic group may
- Heterocycle or “heterocyclyl” means a saturated or unsaturated aromatic or non-aromatic ring, and the aromatic or non-aromatic ring may be a 3 to 10 membered monocyclic ring, a 4 to 12 membered bicyclic ring or a 10 to 15 membered member.
- a tricyclic system comprising 1 to 4 heteroatoms selected from N, O or S, preferably a 3 to 10 membered heterocyclic group, and optionally substituted N, S in the ring of the heterocyclic group can be oxidized to various oxidations state.
- the heterocyclic group may be bonded to a hetero atom or a carbon atom, and the heterocyclic group may be bonded to a bridged or spiro ring, and non-limiting examples include an epoxyethyl group, a glycidyl group, an azacyclopropyl group, and an oxocyclic ring.
- Bn is a benzyl group, that is, a -CH 2 phenyl group.
- ⁇ -adrenergic receptor binding group refers to a group capable of binding to a ⁇ -adrenergic receptor; for example, see review article “ ⁇ -adrenergic receptors in Comprehensive Medicinal Chemistry, 1990, BEMain, p187 (Pergamon Press) ".
- the above-mentioned groups are also described, for example, in WO/2005092841, US/20050215542, WO/2005070872, WO/2006023460, WO/2006051373, WO/2006087315 and WO/2006032627.
- alkyl group optionally substituted by F means that the alkyl group may, but need not, be substituted by F, indicating a case where the alkyl group is substituted by F and a case where the alkyl group is not substituted by F.
- “Pharmaceutical composition” means a mixture of one or more of the compounds described herein or a physiologically/pharmaceutically acceptable salt thereof and other constituents.
- a compound wherein the other components comprise a physiologically/pharmaceutically acceptable carrier and excipient.
- Carrier refers to a carrier or diluent that does not cause significant irritation to the organism and does not abrogate the biological activity and properties of the administered compound.
- Excipient refers to an inert substance that is added to a pharmaceutical composition to further depend on the administration of the compound.
- excipients include, but are not limited to, calcium carbonate, calcium phosphate, various sugars and different types of starch, cellulose derivatives (including microcrystalline cellulose), gelatin, vegetable oils, polyethylene glycols, diluents, Granules, lubricants, binders, disintegrators, and the like.
- Prodrug means a compound that can be converted to a biologically active compound of the invention under physiological conditions or by solvolysis. Prodrugs of the invention are prepared by modifying functional groups in the compounds of the invention which can be removed by conventional procedures or in vivo to provide the parent compound.
- Stepoisomer refers to isomers resulting from the spatial arrangement of atoms in a molecule, including cis and trans isomers, enantiomers and conformational isomers.
- Effective dose refers to an amount of a compound that causes a physiological or medical translation of a tissue, system or subject, which amount is sought, and includes one or more of the conditions or conditions sufficient to prevent treatment when administered to a subject. The amount of a compound that occurs or reduces it to some extent.
- Solvate means a compound of the invention or a salt thereof, which also includes a stoichiometric or non-stoichiometric amount of solvent bound by intermolecular non-covalent forces.
- solvent is water, it is a hydrate.
- IC 50 refers to the half-inhibitory concentration, which is the concentration at which half of the maximum inhibitory effect is achieved.
- the structure of the compound is determined by nuclear magnetic resonance (NMR) or (and) mass spectrometry (MS).
- NMR nuclear magnetic resonance
- MS mass spectrometry
- the NMR shift ( ⁇ ) is given in units of 10 -6 (ppm).
- the NMR was measured using a (Bruker Avance III 400 and Bruker Avance 300) nuclear magnetic apparatus, and the solvent was deuterated dimethyl sulfoxide (DMSO-d 6 ), deuterated chloroform (CDCl 3 ), deuterated methanol (CD 3 OD). ), the internal standard is tetramethylsilane (TMS).
- the measurement of MS was carried out (Agilent 6120B (ESI) and Agilent 6120B (APCI)).
- the HPLC was measured using an Agilent 1260 DAD high pressure liquid chromatograph (Zorbax SB-C18 100 x 4.6 mm).
- Thin layer chromatography silica gel plate uses Yantai Yellow Sea HSGF254 or Qingdao GF254 silica gel plate.
- the specification of silica gel plate used for thin layer chromatography (TLC) is 0.15mm ⁇ 0.20mm.
- the specification for thin layer chromatography separation and purification is 0.4mm. ⁇ 0.5mm.
- Column chromatography generally uses Yantai Huanghai silica gel 200-300 mesh silica gel as a carrier.
- the known starting materials of the present invention may be synthesized by or according to methods known in the art, or may be purchased from Titan Technology, Anike Chemical, Shanghai Demo, Chengdu Kelon Chemical, Suiyuan Chemical Technology, and Belling Technology. And other companies.
- the nitrogen atmosphere means that the reaction flask is connected to a nitrogen balloon having a volume of about 1 L.
- the hydrogen atmosphere means that the reaction flask is connected to a hydrogen balloon of about 1 L volume.
- the hydrogenation reaction is usually evacuated, charged with hydrogen, and operated three times. Unless otherwise stated in the examples, the reaction was carried out under a nitrogen atmosphere.
- the solution means an aqueous solution. There is no particular description in the examples, and the reaction temperature is room temperature.
- the room temperature is an optimum reaction temperature of 20 ° C to 30 ° C. There is no special description in the examples, and M is mol/L.
- TBS is tert-butyldimethylsilyl. Boc is a tert-butyloxycarbonyl group. Bn is a benzyl group.
- TsOH ⁇ H 2 O is p-toluenesulfonic acid monohydrate.
- DMF is N,N-dimethylformamide.
- THF is tetrahydrofuran.
- DCM dichloromethane.
- HATU (2-(7-oxobenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate, CAS: 148893-10-1).
- Step 5 3-[2-[3-(Bromomethyl)phenyl]ethoxy]propanoic acid tert-butyl ester (Intermediate 2)
- Step 5 tert-Butyl 3-[2-[4-(chloromethyl)phenyl]ethoxy]propanoate (Intermediate 4)
- N-butylamine (5a) (14.6 g, 0.2 mol) was added to methanol (150 mL), and aqueous solution of glyoxal-1,1-dimethylacetal (5b) (43.2 g, 0.22 mol, 60% aqueous solution) was added. After stirring at room temperature for 6 hours, sodium borohydride (11.3 g, 0.3 mol) was added portionwise, and the mixture was reacted at room temperature for 2 hours. After concentrating under reduced pressure, the reaction mixture was evaporated, evaporated, evaporated, evaporated, evaporated. 30.0 g, yield 93%).
- Second step 4-[4-(2-Thienyl)pyrimidin-2-yl]-1-oxa-4,9-diazaspiro[5.5]undecane-9-carboxylic acid tert-butyl ester
- Second step 4-[4-phenoxypyrimidin-2-yl]-1-oxa-4,9-diazaspiro[5.5]undecane-9-carboxylic acid tert-butyl ester (intermediate 18 )
- Second step 4-[4-anilinopyrimidin-2-yl]-1-oxa-4,9-diazaspiro[5.5]undecane-9-carboxylic acid tert-butyl ester (Intermediate 19)
- 1-Oxo-4,9-diazaspiro[5.5]undecane-9-carboxylic acid tert-butyl ester (1.28 g, 5.0 mmol) was added to 1,4-dioxane (50 mL) and added 1-Bromo-4-methoxybenzene 25a (1.4 g, 7.5 mmol), sodium tert-butoxide (1.2 g, 12.5 mmol), 4,5-bisdiphenylphosphino-9,9-dimethyloxa ⁇ (0.4 g, 0.7 mmol), palladium acetate (0.08 g, 0.35 mmol).
- 1-Oxo-4,9-diazaspiro[5.5]undecane-9-carboxylic acid tert-butyl ester (1.28 g, 5.0 mmol) was added to 1,4-dioxane (50 mL) and added 1-Bromo-2-chlorobenzene (1.43 g, 7.5 mmol), sodium tert-butoxide (1.2 g, 12.5 mmol), 4,5-bis-diphenylphosphino-9,9-dimethyloxanthene (0.40) g, 0.7 mmol), palladium acetate (0.08 g, 0.35 mmol), and reacted at 100 ° C for 8 hours under a nitrogen atmosphere.
- Example 1 3-[2-[3-[2-[4-[1-(Cyclopropylmethyl)pyrazole-3-carbonyl]-1-oxa-4,9-hetero[5.5] Monoalkane-9-yl]ethyl]phenyl]ethoxy]-N-[2-[[(2R)-2-hydroxy-2-(8-hydroxy-2-oxo-1H-quinoline- 5-yl)ethyl]amino]ethyl]-N-methyl-propionamide (Compound 1)
- Step 5 3-[2-[3-[2-[4-[1-(Cyclopropylmethyl)pyrazole-3-carbonyl]-1-oxa-4,9-diazaspiro[ 5.5] undecyl-9-yl]ethyl]phenyl]ethoxy]propionic acid tert-butyl ester (1F)
- Step 6 3-[2-[3-[2-[4-[1-(Cyclopropylmethyl)pyrazole-3-carbonyl]-1-oxa-4,9-diazaspiro[ 5.5] undecane-9-yl]ethyl]phenyl]ethoxy]propionic acid (1G)
- Step 7 3-[2-[3-[2-[4-[1-(Cyclopropylmethyl)pyrazole-3-carbonyl]-1-oxa-4,9-diazaspiro[ 5.5] undecane-9-yl]ethyl]phenyl]ethoxy]-N-(2,2-dimethoxyethyl)-N-methyl-propionamide (1H)
- Step 8 N-[2-[[(2R)-2-[tert-butyl(dimethyl)silyl]oxy-2-(8-hydroxy-2-oxo-1H-quinoline-5) -yl)ethyl]amino]ethyl-3-[2-[3-[2-[4-[1-(cyclopropylmethyl)pyrazole-3-carbonyl]-1-oxa-4 ,9-diazaspiro[5.5]undec-9-yl]ethyl]phenyl]ethoxy]-N-(2,2-dimethoxyethyl)-N-methyl-prop Amide (1I)
- the ninth step 3-[2-[3-[2-[4-[1-(cyclopropylmethyl)pyrazole-3-carbonyl]-1-oxa-4,9-hetero[5.5] Monoalkane-9-yl]ethyl]phenyl]ethoxy]-N-[2-[[(2R)-2-hydroxy-2-(8-hydroxy-2-oxo-1H-quinoline- 5-yl)ethyl]amino]ethyl]-N-methyl-propionamide (Compound 1)
- Example 2 3-[2-[3-[2-[4-[1-(Cyclopropylmethyl)pyrazole-3-carbonyl]-1-oxa-4,9-diazaspiro[5.5 Undecyl-9-yl]ethyl]phenyl]ethoxy]-N-[2-[[(2R)-2-hydroxy-2-(5-hydroxy-3-oxo-4H-1) , 4-benzoxazine-8-yl)ethyl]amino]ethyl]-N-methyl-propionamide (Compound 2)
- Step 6 N-[2-[[(2R)-2-[tert-butyl(dimethyl)silyl]oxy-2-(8-hydroxy-2-oxo-1H-quinoline-5) -yl)ethyl]amino]ethyl-N-methyl-3-[2-[3-[[4-(2-pyridyl)-1-oxa-4,9-diazaspiro[ 5.5] undecane-9-yl]methyl]phenyl]ethoxy]propanamide (3G)
- Step 7 N-[2-[[(2R)-2-hydroxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]- N-methyl-3-[2-[3-[[4-(2-pyridyl)-1-oxa-4,9-diazaspiro[5.5]undec-9-yl]methyl Phenyl]ethoxy]propanamide (compound 3)
- 3C trifluoroacetate (0.5 g, 1.5 mmol) was added to EtOAc (15 mL).
- Intermediate 3 (0.55 g, 1.5 mmol), potassium carbonate (1.1 g, 7.5 mmol) and water (0.5 mL) Stir at 60 ° C for 24 hours.
- water 50 mL was added, and ethyl acetate (100 mL ⁇ 2) was evaporated.
- Step 5 3-[2-[4-[[4-(2-propynylthiazole-4-carbonyl)-1-oxa-4,9-diazaspiro[5.5]undecane-9 -yl]methyl]phenyl]ethoxy]propionic acid tert-butyl ester (5F)
- Step 6 3-[2-[4-[[4-(2-propynylthiazole-4-carbonyl)-1-oxa-4,9-diazaspiro[5.5]undecane-9 -yl]methyl]phenyl]ethoxy]propionic acid (5G)
- Step 7 N-(2,2-Dimethoxyethyl)-N-methyl-3-[2-[4-[[4-(2-propynylthiazole-4-carbonyl)-1 -oxa-4,9-diazaspiro[5.5]undec-9-yl]methyl]phenyl]ethoxy]propanamide (5H)
- Step 8 N-[2-[[(2R)-2-[tert-butyl(dimethyl)silyl]oxy-2-(8-hydroxy-2-oxo-1H-quinoline-5) -yl)ethyl]amino]ethyl-N-methyl-3-[2-[4-[[4-(2-propynylthiazole-4-carbonyl)-1-oxa-4,9- Diazaspiro[5.5]undecyl-9-yl]methyl]phenyl]ethoxy]propanamide (5I)
- Step 9 N-[2-[[(2R)-2-hydroxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]- N-methyl-3-[2-[4-[[4-(2-propynylthiazole-4-carbonyl)-1-oxa-4,9-diazaspiro[5.5]undecane- 9-yl]methyl]phenyl]ethoxy]propanamide (compound 5)
- Example 7 3-[2-[3-[2-[4-(3-ethynylbenzoyl)-1-oxa-4,9-diazaspiro[5.5]undecane-9- Ethyl]ethyl]phenyl]ethoxy]-N-[2-[[(2R)-2-hydroxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)) Amino]ethyl]-N-methyl-propionamide (compound 7)
- the third step 3-[2-[3-[2-[4-(3-ethynylbenzoyl)-1-oxa-4,9-diazaspiro[5.5]undecane-9- Tert-butyl]ethyl [ethyl]phenyl]ethoxy]propanoate (7D)
- the fifth step N-(2,2-dimethoxyethyl)-3-[2-[3-[2-[4-(3-ethynylbenzoyl)-1-oxa-4, 9-diazaspiro[5.5]undecane-9-yl]ethyl]phenyl]ethoxy]-N-methyl-propionamide (7F)
- Step 6 N-[2-[[(2R)-2-[tert-butyl(dimethyl)silyl]oxy-2-(8-hydroxy-2-oxo-1H-quinoline-5) -yl)ethyl]amino]ethyl]-3-[2-[3-[2-[4-(3-ethynylbenzoyl)-1-oxa-4,9-diaza snail [5.5] undecane-9-yl]ethyl]phenyl]ethoxy]-N-methyl-propionamide (7G)
- Step 7 3-[2-[3-[2-[4-(3-ethynylbenzoyl)-1-oxa-4,9-diazaspiro[5.5]undecane-9- Ethyl]ethyl]phenyl]ethoxy]-N-[2-[[(2R)-2-hydroxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)) Amino]ethyl]-N-methyl-propionamide (compound 7)
- Example 8 3-[2-[3-[2-[4-(1,2-benzothiazol-3-yl)-1-oxa-4,9-diazaspiro[5.5] eleven Alkan-9-yl]ethyl]phenyl]ethoxy]-N-butyl-N-[2-[[(2R)-2-hydroxy-2-(8-hydroxy-2-oxo-1H) -quinolin-5-yl)ethyl]amino]ethyl]propanamide (compound 8)
- the third step 3-[2-[3-[2-[4-(1,2-benzothiazol-3-yl)-1-oxa-4,9-diazaspiro[5.5] eleven Tert-butyl 9-yl]ethyl]phenyl]ethoxy]propanoate (8D)
- the fifth step 3-[2-[3-[2-[4-(1,2-benzothiazol-3-yl)-1-oxa-4,9-diazaspiro[5.5] eleven Alkan-9-yl]ethyl]phenyl]ethoxy]-N-butyl-(2,2-dimethoxyethyl)propanamide (8F)
- the sixth step 3-[2-[3-[2-[4-(1,2-benzothiazol-3-yl)-1-oxa-4,9-diazaspiro[5.5] eleven Alkan-9-yl]ethyl]phenyl]ethoxy]-N-butyl-N-[2-[[(2R)-2-[tert-butyl(dimethyl)silyl]oxy-2 -(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]propanamide (8G)
- the seventh step 3-[2-[3-[2-[4-(1,2-benzothiazol-3-yl)-1-oxa-4,9-diazaspiro[5.5] eleven Alkan-9-yl]ethyl]phenyl]ethoxy]-N-butyl-N-[2-[[(2R)-2-hydroxy-2-(8-hydroxy-2-oxo-1H) -quinolin-5-yl)ethyl]amino]ethyl]propanamide (compound 8)
- 3C trifluoroacetic acid salt (0.46 g, 2.0 mmol) was added to EtOAc (25 mL).
- Example 11 3-[2-[3-[2-[4-(4-ethylpyrimidin-2-yl)-1-oxa-4,9-diazaspiro[5.5]undecane -9-yl]ethyl]phenyl]ethoxy]-N-[2-[[(2R)-2-hydroxy-2-(8-hydroxy-2-oxo-1H-quinoline-5-) Ethyl]amino]ethyl]-N-methyl-propionamide (compound 11)
- Step 6 N-[2-[[(2R)-2-[tert-butyl(dimethyl)silyl]oxy-2-(8-hydroxy-2-oxo-1H-quinoline-5-) Ethyl]amino]amino][3-[3-[[4-(4-ethylpyrimidin-2-yl)-1-oxa-4,9-diazaspiro[ 5.5] undecane-9-yl]ethyl]phenyl]ethoxy]-N-methyl-propionamide (11F)
- Step 7 3-[2-[3-[2-[4-(4-ethylpyrimidin-2-yl)-1-oxa-4,9-diazaspiro[5.5]undecane -9-yl]ethyl]phenyl]ethoxy]-N-[2-[[(2R)-2-hydroxy-2-(8-hydroxy-2-oxo-1H-quinoline-5-) Ethyl]amino]ethyl]-N-methyl-propionyl (Compound 11)
- the third step 3-[2-[3-[[4-(4-methylpyrimidin-2-yl)-1-oxa-4,9-diazaspiro[5.5]undecane-9- Tert-butyl]ethyl]ethyl]phenyl]ethoxy]propanoate (12C)
- Step 5 N-(2,2-Dimethoxyethyl)-N-methyl-3-[2-[3-[[4-(4-methylpyrimidin-2-yl)-1- Oxa-4,9-diazaspiro[5.5]undecyl-9-yl]ethyl]phenyl]ethoxy]-propionamide (12E)
- Step 6 N-[2-[[(2R)-2-[tert-butyl(dimethyl)silyl]oxy-2-(8-hydroxy-2-oxo-1H-quinoline-5-) Ethyl]amino]amino]ethyl-N-methyl-3-[2-[3-[[4-(4-methylpyrimidin-2-yl)-1-oxa-4,9- Diazaspiro[5.5]undecyl-9-yl]ethyl]phenyl]ethoxy]propanamide (12F)
- Step 7 N-[2-[[(2R)-2-hydroxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]- N-methyl-3-[2-[3-[2-[4-(4-methylpyrimidin-2-yl)-1-oxa-4,9-diazaspiro[5.5] eleven Alkan-9-yl]ethyl]phenyl]ethoxy]propanamide (Compound 12)
- Example 13 3-[2-[4-[2-[4-[1-(Cyclopropylmethyl)pyrazole-3-carbonyl]-1-oxa-4,9-diazaspiro[5.5 Undecyl-9-yl]ethyl]phenyl]ethoxy]-N-[2-[[(2R)-2-hydroxy-2-(8-hydroxy-2-oxo-1H-quinaline ⁇ -5-yl)ethyl]amino]ethyl]-N-methyl-propionamide (compound 13)
- the third step 3-[2-[4-[2-[4-[1-(cyclopropylmethyl)pyrazole-3-carbonyl]-1-oxa-4,9-diazaspiro[5.5 Undecyl-9-yl]ethyl]phenyl]ethoxy]propionic acid (13C)
- Step 5 N-[2-[[(2R)-2-[tert-butyl(dimethyl)silyl]oxy-2-(8-hydroxy-2-oxo-1H-quinoline-5-) Ethyl]amino]ethyl-3-[2-[4-[2-[4-[1-(cyclopropylmethyl)pyrazole-3-carbonyl]-1-oxa-4, 9-diazaspiro[5.5]undecyl-9-yl]ethyl]phenyl]ethoxy]-N-(2,2-dimethoxyethyl)-N-methyl-propionamide (13E)
- Step 6 3-[2-[4-[2-[4-[1-(Cyclopropylmethyl)pyrazole-3-carbonyl]-1-oxa-4,9-diazaspiro[5.5 Undecyl-9-yl]ethyl]phenyl]ethoxy]-N-[2-[[(2R)-2-hydroxy-2-(8-hydroxy-2-oxo-1H-quinaline ⁇ -5-yl)ethyl]amino]ethyl]-N-methyl-propionamide (compound 13)
- the 14A (0.50g, 0.75mmol) was dissolved in THF (10mL) was added TsOH ⁇ H 2 O (0.71g, 3.7mmol), stirred for 40 °C 1 hour a solution of saturated aqueous sodium bicarbonate (50mL) The reaction was quenched The extract was extracted with DCM (100 mL ⁇ 2). The combined organic layer was dried over anhydrous sodium sulfate.
- Step 5 N-butyl-N-(2,2-dimethoxyethyl)-3-[2-[3-[2-[4-(1-methylpyrazol-3-yl)) -1-oxa-4,9-diazaspiro[5.5]undec-9-yl]ethyl]phenyl]ethoxy]propanamide (15F)
- Step 6 N-butyl-N-[2-[[(2R)-2-[tert-butyl(dimethyl)silyl]oxy-2-(8-hydroxy-2-oxo-1H-) Quinoline-5-yl)ethyl]amino]ethyl]-3-[2-[3-[2-[4-(1-methylpyrazol-3-yl)-1-oxa-4 ,9-diazaspiro[5.5]undecane-9-yl]ethyl]phenyl]ethoxy]-propionamide (15G)
- Step 7 N-butyl-N-[2-[[(2R)-2-hydroxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino) ]ethyl]-3-[2-[3-[2-[4-(1-methylpyrazol-3-yl)-1-oxa-4,9-diazaspiro[5.5] eleven Alkan-9-yl]ethyl]phenyl]ethoxy]propanamide (compound 15)
- Example 16 3-[2-[3-[2-[4-(4-ethyl-2-pyridyl)-1-oxa-4,9-diazaspiro[5.5]undecane- 9-yl]ethyl]phenyl]ethoxy]-N-[2-[[(2R)-2-hydroxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl) Ethyl]amino]ethyl]-N-methyl-propionamide (Compound 16)
- the third step 4-(4-ethyl-2-pyridine)-1-oxa-4,9-diazaspiro[5.5]undecane (16D)
- 16D trifluoroacetic acid salt (0.52 g, 1.4 mmol) was taken in EtOAc (15 mL).
- Intermediate 1 (0.49 g, 1.4 mmol), potassium carbonate (0.95 g, 6.9 mmol), water (0.5 mL) ), stirring at 60 ° C for 24 hours.
- water 50 mL was added, and ethyl acetate (100 mL ⁇ 2) was evaporated.
- Step 5 3-[2-[3-[2-[4-(4-ethyl-2-pyridyl)-1-oxa-4,9-diazaspiro[5.5]undecane- 9-yl]ethyl]phenyl]ethoxy]propionic acid (16F)
- Step 6 N-(2,2-Dimethoxyethyl)-3-[2-[3-[2-[4-(4-ethyl-2-pyridyl)-1-oxa-- 4,9-diazaspiro[5.5]undecane-9-yl]ethyl]phenyl]ethoxy]-N-methyl-propionamide (16G)
- Step 7 N-[2-[[(2R)-2-[tert-butyl(dimethyl)silyl]oxy-2-(8-hydroxy-2-oxo-1H-quinoline-5-) Ethyl]amino]amino]ethyl]-3-[2-[3-[2-[4-(4-ethyl-2-pyridyl)-1-oxa-4,9-diaza Spirulin [5.5] undecane-9-yl]ethyl]phenyl]ethoxy]-N-methyl-propionamide (16H)
- Step 7 3-[2-[3-[2-[4-(4-ethyl-2-pyridyl)-1-oxa-4,9-diazaspiro[5.5]undecane- 9-yl]ethyl]phenyl]ethoxy]-N-[2-[[(2R)-2-hydroxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl) Ethyl]amino]ethyl]-N-methyl-propionamide (Compound 16)
- 17C trifluoroacetic acid salt (0.303 g, 0.69 mmol) was dissolved in DCM (20 mL), then triethylamine (0.25 g, 3..45 mmol), methylaminoacetal dimethylacetal (0.098 g, 0.82) Mmol), HATU (0.393 g, 1.04 mmol), stirred at room temperature for 2 h.
- the aqueous solution of saturated sodium hydrogencarbonate (50 mL) was added dropwise, and the mixture was evaporated, evaporated, evaporated.
- the fifth step N-[2-[[(2R)-2-hydroxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]- N-methyl-3-[2-[4-[[4-(2-pyridyl-1-oxa-4,9-diazaspiro[5.5]undec-9-yl]methyl) Phenyl]ethoxy]propanamide (compound 17)
- Example 18 3-[2-[3-[[4-(4-ethyl-2-pyridyl)-1-oxa-4,9-heterooxadia[5.5]undecane-9- Methyl]phenyl]ethoxy]-N-[2-[[(2R)-2-hydroxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)) Amino]ethyl]-N-methyl-propionamide (compound 18)
- Step 5 3-[2-[3-[[4-(4-ethyl-2-pyridyl)-1-oxa-4,9-diazaspiro[5.5]undecane-9- Methyl]phenyl]ethoxy]-N-[2-[[(2R)-2-hydroxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)) Amino]ethyl]-N-methyl-propionamide (compound 18)
Landscapes
- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Animal Behavior & Ethology (AREA)
- General Health & Medical Sciences (AREA)
- Veterinary Medicine (AREA)
- Public Health (AREA)
- Life Sciences & Earth Sciences (AREA)
- Medicinal Chemistry (AREA)
- Pharmacology & Pharmacy (AREA)
- Pulmonology (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- General Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Epidemiology (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
本发明涉及一种二氮杂螺[5.5]十一碳烷衍生物及其用途,该衍生物如通式(I)所示的化合物或其立体异构体、水合物、代谢产物、溶剂化物、药学上可接受的盐、共晶或前药,本发明还涉及所述衍生物的制备方法和在制备用于治疗气道阻塞性疾病药物中的应用,其中,各取代基的定义与说明书中一致。
Description
本发明涉及一种二氮杂螺[5.5]十一碳烷衍生物及其制备方法和在医药上的应用,具体是一种具有蕈毒碱受体拮抗和β2-肾上腺素能受体激动的双重活性的新颖二氮杂螺[5.5]十一碳烷或其立体异构体、水合物、溶剂化物、代谢产物、药学上可接受的盐、共晶或前药、其药物组合物以及其在医药上的应用。
支气管扩张剂对于呼吸疾病例如慢性阻塞性肺疾病(COPD)及哮喘等的治疗起重要作用。临床中广泛使用的支气管扩张剂包括蕈毒碱受体拮抗剂和β2-肾上腺素能激动剂。蕈毒碱受体拮抗剂通过降低气道平滑肌的迷走神经胆碱能水平来发挥支气管扩张的效力。目前所使用的吸入蕈毒碱受体拮抗剂包括异丙托溴铵、氧托溴铵、格隆溴铵、噻托溴铵、阿地溴胺和芜地溴胺。β2-肾上腺素能激动剂通过刺激气道平滑肌的肾上腺素能受体而使支气管扩张,逆转支气管收缩剂对各种介质如乙酰胆碱的反应。目前所使用的β2-肾上腺素能激动剂包括沙丁胺醇、沙美特罗、阿福特罗、福美特罗、维兰特罗和茚达特罗。这些药物除了改善肺的功能,也可改善患者生活质量并减少病情恶化。
随着更多的临床研究发现,证明联合使用蕈毒碱受体拮抗剂和β2-肾上腺素能激动剂比单独使用其中一种治疗剂更有效,目前临床上将蕈毒碱受体拮抗剂和β2-肾上腺素能激动剂制备成复方制剂,用于哮喘和中重度COPD的治疗,这类复方制剂主要包括Anoro Ellipta(芜地溴胺/维兰特罗)、Ultibro Breezhaler(格隆溴铵/茚达特罗)和异丙托溴胺/沙丁胺醇等。虽然复方制剂比其中单一制剂具有更好的治疗效果,但是在制剂制备上有更高的要求。
因此,人们希望开发同时具有蕈毒碱受体拮抗和β2-肾上腺素能激动双重作用的药物,这种双功能药物具有两种成分组合的药学优点,同时具备单一的分子药代动力学。这些化合物以单一治疗剂的形式给药,可以由两种不同且可能协同起效的作用模式提供支气管扩张作用。另外,具有蕈毒碱受体拮抗和β2-肾上腺素能激动双重作用(MABA)的化合物还可以与皮质类固醇(ICS)消炎剂药物组合,形成两种治疗剂(MABA/ICS)而提供三重作用的治疗效果(Expert Opin.Investig.Drugs(2014)23(4):453-456)。
因此,有必要开发新颖的同时具有蕈毒碱受体拮抗和β2-肾上腺素能激动的双重活性药物,以提供更有效的单一治疗剂量或者复方制剂,为患者提供更多的临床用药选择。
发明内容
本发明提供一种通式(I)所示的化合物或其立体异构体、水合物、代谢产物、溶剂化物、药学上可接受的盐、共晶或前药:
其中:
R1选自C6-12碳环或者5至12元的杂环,所述的碳环或者杂环任选进一步被0至5个R1a取代,且
所述的杂环含有1至4个选自N、O或S的杂原子;
R1a选自F、Cl、Br、I、-(=O)、硝基、氰基、羟基、羧基、-C(=O)OC1-6烷基、氨基、C1-6烷基、C2-6炔基、C1-6烷氧基、C3-12碳环、5至12元的杂环、-O-C3-12碳环、-NH-C3-12碳环或-CH2-C3-6环烷基,所述的羟基、氨基、烷基、炔基、烷氧基、碳环、杂环或环烷基任选进一步被0至4个选自F、Cl、Br、I、氰基、羟基、C1-6烷基、C2-6炔基、C1-4烷氧基、C3-6环烷基或C3-10碳环或5至10元的杂环的取代基所取代,所述的杂环含有1至4个选自N、O或S的杂原子;
W选自键或-C(=O)-;
条件是W选自-C(=O)-时,R1被1至5个R1a取代,且至少有1个R1a选自C2-6炔基或-CH2-C3-6环烷基;
R2、R3、R4、R5各自独立的选自C1-6亚烷基,所述的亚烷基任选进一步被0至4个选自F、Cl、Br、I、氰基、羟基、C1-6烷基、C1-6烷氧基、苯基或苯基-C1-4亚烷基的取代基所取代;
A选自C3-12碳环或5至12元杂环,所述的碳环或杂环任选进一步被0至5个F、Cl、Br、I、氰基、羟基、氨基、C1-6烷基、C2-6炔基、C1-4烷氧基或C3-6环烷基的取代基取代,且所述的杂环含有1至3个选自N、O或S的杂原子;
X1和X2各自独立的选自键、-O-、-C(=O)-、-C(=O)O-、-OC(=O)-、-S-、-S(=O)-、-S(=O)2-、-C(=O)NRx-、-NRxC(=O)-、-OC(=O)NRx-、-NRxC(=O)O-、-NRxC(=O)NRx、-NRxS(=O)2-、-S(=O)2NRx-、-NRxS(=O)2NRx或-NRx-;
Rx各自独立选自H、C1-6烷基或C3-8环烷基,所述的烷基或环烷基任选进一步被0至5个选自F、Cl、Br、I、C1-4烷基、C3-6环烷基或C1-4烷氧基的取代基所取代;
作为选择R3-X1-R4-X2-R5可为亚乙基;
R6、R7各自独立的选自H或C1-4烷基;
R8选自H或OH;
本发明优选方案,一种通式(I)所示的化合物或其立体异构体、水合物、代谢产物、溶剂化物、药学上可接受的盐、共晶或前药,其中:
B优选R10、R11、R12、R13、R14、R15、R16、R17或R18各自独立的选自H、F、Cl、Br、I、CF3、OH、-CH2OH、氰基、羧基、C1-4烷基、C1-4烷氧基、-C(=O)C1-4烷基、-C(=O)OC1-4烷基、-NHC(=O)H、-NHS(=O)2-C1-4烷基、-NHS(=O)2-NH2或-NHS(=O)2-NHC1-4烷基,Q选
自-CRq1=CRq2-、-CRq1Rq2CRq3Rq4-、-O-、-S-、-OCRq1Rq2-、-CRq1Rq2O-、-SCRq1Rq2-、-CRq1Rq2S-,所述Rq1、Rq2、Rq3或Rq4各自独立的选自选自H、F、Cl、Br、I或C1-4烷基;
本发明优选方案,一种通式(I)所示的化合物或其立体异构体、水合物、代谢产物、溶剂化物、药学上可接受的盐、共晶或前药,其中:
R1选自C6-10碳环或者5至10元的杂环,优选苯环、噻吩环、呋喃环、吡咯环、噻唑环、噁唑环、咪唑环、异噻唑环、异噁唑环、吡唑环、吡啶环、喹啉环、异喹啉环、苯并异噻唑环、吲唑环、1H-吡唑[4,3-b]吡啶环或所述的碳环、杂环、苯环、噻吩环、呋喃环、吡咯环、噻唑环、噁唑环、咪唑环、异噻唑环、异噁唑环、吡唑环、吡啶环、喹啉环、异喹啉环、苯并异噻唑环、吲唑环、1H-吡唑[4,3-b]吡啶环或任选进一步被0至5个R1a取代,且所述的杂环含有1至4个选自N、O或S的杂原子;
R1a选自F、Cl、Br、-(=O)、硝基、氰基、羟基、羧基、-C(=O)OC1-4烷基、氨基、C1-4烷基、C2-4炔基、C1-4烷氧基、C3-10碳环、5至10元的杂环、-O-C3-10碳环、-NH-C3-10碳环或-CH2-C3-6环烷基,所述的羟基、氨基、烷基、炔基、烷氧基、碳环、杂环或环烷基任选进一步被0至4个选自F、Cl、Br、氰基、羟基、C1-4烷基、C1-4烷氧基、C3-6环烷基、C3-10碳环或5至10元的杂环的取代基所取代,所述的杂环含有1至4个选自N、O或S的杂原子;
W选自键或-C(=O)-;
条件是W选自-C(=O)-时,R1被1至5个R1a取代,且至少有1个R1a选自C2-4炔基或-CH2-C3-6环烷基;
R2、R5各自独立的选自C1-4亚烷基,所述的亚烷基任选进一步被0至4个选自F、甲基、乙基、甲氧基或乙氧基的取代基所取代;
R3、R4各自独立的选自C1-4亚烷基,优选亚甲基、亚乙基、亚丙基、-CH2CH(CH3)-、-CH(CH3)CH2-、-C(CH3)2CH2-、-CH2C(CH3)2-、亚丁基、-CH(CH3)CH2CH2-、-CH2CH(CH3)CH2-或-CH2CH(CH3)CH2-,所
述的亚烷基、亚甲基、亚乙基、亚丙基或亚丁基任选进一步被0至4个选自F、甲基、乙基、丙基、异丙基、丁基、甲氧基、乙氧基、苯基、苯基-亚甲基、苯基-亚乙基、苯基-亚丙基或苯基-亚丁基的取代基所取代;
A选自C6-10碳环或5至10元杂环,优选所述的碳环、杂环、
任选进一步被0至5个F、Cl、Br、I、氰基、羟基、氨基、C1-6烷基、C2-6炔基、C1-4烷氧基或C3-6环烷基的取代基取代,且所述的杂环含有1至3个选自N、O或S的杂原子;
X1和X2各自独立的选自键、-O-、-C(=O)-、-C(=O)O-、-OC(=O)-、-S-、-S(=O)-、-S(=O)2-、-C(=O)NRx-、-NRxC(=O)-、-OC(=O)NRx-、-NRxC(=O)O-、-NRxC(=O)NRx、-NRxS(=O)2-、-S(=O)2NRx-、-NRxS(=O)2NRx或-NRx-,优选键、-O-、-C(=O)-、-C(=O)O-、-OC(=O)-、-C(=O)NRx-、-NRxC(=O)-、-OC(=O)NRx-、-NRxC(=O)O-或-NRx-;
Rx各自独立选自H、C1-6烷基或C3-8环烷基,所述的烷基或环烷基任选进一步被0至5个选自F、Cl、Br、I、甲基、乙基、环丙基、环丁基、环戊基、环己基、甲氧基或乙氧基的取代基所取代;
Rx优选H、甲基、乙基、丙基、异丙基、丁基、异丁基、戊基、环丙基、环丁基、环戊基、环己基、-CH2-环丙基、-CH2-环丁基或-CH2-环戊基;
作为选择R3-X1-R4-X2-R5可为亚乙基;
R6、R7各自独立的选自H或C1-4烷基,优选H、甲基或乙基;
R8选自H或OH;
本发明优选方案,一种通式(I)所示的化合物或其立体异构体、水合物、代谢产物、溶剂化物、药学上可接受的盐、共晶或前药,其中:
R1选自苯环、噻吩环、呋喃环、吡咯环、噻唑环、噁唑环、咪唑环、异噻唑环、异噁唑环、吡唑环、吡啶环、嘧啶环、吡嗪环、喹啉环、异喹啉环、苯并异噻唑环、吲唑环、1H-吡唑[4,3-b]吡啶环或
所述的苯环、噻吩环、呋喃环、吡咯环、噻唑环、噁唑环、咪唑环、异噻唑环、异噁唑环、吡唑环、吡啶环、嘧啶环、吡嗪环、喹啉环、异喹啉环、苯并异噻唑环、吲唑环、1H-吡唑[4,3-b]吡啶环或任选进一步被0至5个选自R1a取代;
R1a选自F、Cl、Br、硝基、氨基、氰基、羧基、-C(=O)OBn、-CF3、羟基、甲基、乙基、丙基、异丙基、丁基、异丁基、叔丁基、乙炔基、丙炔基、炔丙基、甲氧基、乙氧基、丙氧基、异丙氧基、羟甲基、羟乙基、-C(CH3)2OH、苯氧基、苯胺基、苯基、2-噻吩基、环丙基、环丁基、环戊基、环己基或-CH2-环丙基;
W选自键或-C(=O)-;
条件是W选自-C(=O)-时,R1被1至5个R1a取代,且至少有1个R1a选自乙炔基、丙炔基、炔丙基或-CH2-环丙基;
R2、R5各自独立的选自C1-4亚烷基,所述的亚烷基任选进一步被0至4个选自F、甲基、乙基、甲氧基或乙氧基的取代基所取代;
R2、R5优选亚甲基、亚乙基、亚丙基、-CH2CH(CH3)-、-CH(CH3)CH2-、-C(CH3)2CH2-、-CH2C(CH3)2-、亚丁基、-CH(CH3)CH2CH2-、-CH2CH(CH3)CH2-或-CH2CH(CH3)CH2-;
R3、R4各自独立的选自亚甲基、亚乙基、亚丙基或亚丁基,所述的亚甲基、亚乙基、亚丙基或亚丁基任选进一步被0至4个选自F、甲基、乙基、丙基、异丙基、丁基、甲氧基、乙氧基、苯基、苯基-亚甲基、苯基-亚乙基、苯基-亚丙基或苯基-亚丁基的取代基所取代;
X1和X2各自独立的选自键、-O-、-C(=O)-、-C(=O)O-、-OC(=O)-、-C(=O)NRx-、-NRxC(=O)-、-OC(=O)NRx-、-NRxC(=O)O-或-NRx-;
Rx各自独立选自H、甲基、乙基、丙基、异丙基、丁基、异丁基、戊基、环丙基、环丁基、环戊基、环己基、-CH2-环丙基、-CH2-环丁基或-CH2-环戊基;
作为选择R3-X1-R4-X2-R5可为亚乙基;
R6、R7各自独立的选自H、甲基或乙基;
本发明优选方案,一种通式(II)或式(III)所示的化合物或其立体异构体、水合物、代谢产物、溶剂化
物、药学上可接受的盐、共晶或前药,其中:
式(II)或式(III)中:
p1选自0或1;
p2选自0、1、2、3或4;
Ra选自H、F、Cl、Br、I、C1-4烷基、C1-4烷氧基或C3-6环烷基;
R1选自C6-10碳环或者5至10元的杂环,所述的碳环或者杂环任选进一步被0至5个R1a取代,且所述的杂环含有1至4个选自N、O或S的杂原子;
R1a选自F、Cl、Br、-(=O)、硝基、氰基、羟基、氨基、羧基、-C(=O)OC1-4烷基、C1-4烷基、C2-4炔基、C1-4烷氧基、C3-10碳环、5至10元的杂环、-O-C3-10碳环、-NH-C3-10碳环或-CH2-C3-6环烷基,所述的羟基、氨基、烷基、炔基、烷氧基、碳环、杂环或环烷基任选进一步被0至4个选自F、Cl、Br、氰基、羟基、C1-4烷基、C1-4烷氧基、C3-6环烷基、C3-10碳环或5至10元的杂环的取代基所取代,所述的杂环含有1至4个选自N、O或S的杂原子;
W选自键或-C(=O)-;
条件是W选自-C(=O)-时,R1被1至5个R1a取代,且至少有1个R1a选自C2-4炔基或-CH2-C3-6环烷基;
Rx各自独立选自H、C1-6烷基或C3-8环烷基,所述的烷基或环烷基任选进一步被0至5个选自F、Cl、Br、I、甲基、乙基、环丙基、环丁基、环戊基、环己基、甲氧基或乙氧基的取代基所取代;
Rx优选H、甲基、乙基、丙基、异丙基、丁基、异丁基、戊基、环丙基、环丁基、环戊基、环己基、-CH2-环丙基、-CH2-环丁基或-CH2-环戊基;
R8选自H或羟基;
Q选自-CH=CH-、-CH2CH2-、-O-、-S-、-CH2O-、-OCH2-、-C(CH3)2O-或-OC(CH3)2-;
本发明优选方案,一种通式(II)或式(III)所示的化合物或其立体异构体、水合物、代谢产物、溶剂化物、药学上可接受的盐、共晶或前药,其中:
p1选自0或1;
p2选自0、1、2、3或4;
R1选自苯环、噻吩环、呋喃环、吡咯环、噻唑环、噁唑环、咪唑环、异噻唑环、异噁唑环、吡唑环、吡啶环、吡嗪环、嘧啶环、喹啉环、异喹啉环、苯并异噻唑环、吲唑环、1H-吡唑[4,3-b]吡啶环或所述的苯环、噻吩环、呋喃环、吡咯环、噻唑环、噁唑环、咪唑环、异噻唑环、异噁唑环、吡唑环、吡啶环、嘧啶环、吡嗪环、喹啉环、异喹啉环、苯并异噻唑环、吲唑环、1H-吡唑[4,3-b]吡啶环或任选进一步被0至5个R1a取代;
R1a选自F、Cl、Br、硝基、氨基、氰基、羟基、羧基、-C(=O)OBn、-CF3、甲基、乙基、丙基、异丙基、丁基、异丁基、叔丁基、乙炔基、丙炔基、炔丙基、甲氧基、乙氧基、丙氧基、异丙氧基、羟甲基、羟乙基、-C(CH3)2OH、苯氧基、苯胺基、苯基、2-噻吩基、环丙基、环丁基、环戊基、环己基或-CH2-环丙基;
条件是W选自-C(=O)-时,R1被1至5个R1a取代,且至少有1个R1a选自乙炔基、丙炔基、炔丙基、或-CH2-环丙基;
Ra选自F、Cl、Br、I、甲基、乙基、甲氧基、乙氧基或环丙烷基;
Rx选自H、甲基、乙基、丙基、异丙基、丁基、异丁基、戊基、环丙基、环丁基、环戊基、环己基、-CH2-环丙基、-CH2-环丁基或-CH2-环戊基;
本发明优选方案,一种通式(II)或式(III)所示的化合物或其立体异构体、水合物、代谢产物、溶剂化物、药学上可接受的盐、共晶或前药,其中:
p1选自0或1;
p2选自0、1、2、3或4;
Ra选自F、Cl、Br、I、甲基、乙基、甲氧基、乙氧基或环丙烷基;
Rx选自H、甲基、乙基、丙基、异丙基、丁基、异丁基、戊基、环丙基、环丁基、环戊基、环己基、-CH2-环丙基、-CH2-环丁基或-CH2-环戊基;
本发明优选方案,本发明所涉及的化合物包括,但不限于如下化合物之一:
本发明还涉及提供一种药物组合物,所述的药物组合物含有治疗有效剂量的通式(I)、(II)或(III)任一项所述的化合物或其立体异构体、水合物、代谢产物、溶剂化物、药学上可接受的盐、共晶或前药,以及药学上可接受的载体、稀释剂、佐剂、媒介物或赋形剂;所述的组合物还可进一步包括一种或多种其他治疗剂;优选的,其中所述其他治疗剂选自PDE4抑制剂、M受体拮抗剂、皮质类固醇和β-肾上腺素受体激动剂中的一种或多种。
本发明还涉及提供通式(I)、(II)或(III)所述的化合物或其立体异构体、水合物、代谢产物、溶剂化物、药学上可接受的盐、共晶或前药,或前述药物组合物在制备用于治疗气道阻塞性疾病的药物中的应用,优选的,在制备用于治疗哮喘、慢性阻塞性肺疾病或支气管炎的药物中的应用。
本发明还提供一种治疗气道阻塞性疾病的方法,所述方法包括给药通式(I)、(II)或(III)所述的化合物或其立体异构体、水合物、代谢产物、溶剂化物、药学上可接受的盐、共晶或前药,或上述药物组合物,优选地,所述的气道阻塞性疾病选自哮喘、慢性阻塞性肺疾病或支气管炎。
本发明还提供一种制备式(I)所示化合物的中间体,该中间体选自式(M-1)、式(M-2)或式(M-3)所示化合物或者其立体异构体和药学上可接受的盐:
其中:
Rm1选自氨基保护基或H;
Rm2选自-COOH、-COOC1-6烷基、-X2-R5-OH、-X2-R5-离去基团、-X2-Rm3CHO、-X2-Rm3C(OC1-6烷基)2或所述烷基任选进一步被0至4个F、Cl、Br或C1-4烷基所取代;
Rm3选自较R5少一个碳原子的亚烷基;
R1选自C6-12碳环或5至12元的杂环,优选C6-10碳环或者5至10元的杂环,更优选苯环、噻吩环、呋喃环、吡咯环、噻唑环、噁唑环、咪唑环、异噻唑环、异噁唑环、吡唑环、吡啶环、嘧啶环、吡嗪环、喹啉环、异喹啉环、苯并异噻唑环、吲唑环、1H-吡唑[4,3-b]吡啶环或所述的碳环、杂环、苯环、噻吩环、呋喃环、吡咯环、噻唑环、噁唑环、咪唑环、异噻唑环、异噁唑环、吡唑环、吡啶环、嘧啶环、吡嗪环、喹啉环、异喹啉环、苯并异噻唑环、吲唑环、1H-吡唑[4,3-b]吡啶环或任选进一步被0至5个R1a取代,且所述的杂环含有1至4个选自N、O或S的杂原子;
R1a选自F、Cl、Br、I、-(=O)、硝基、氰基、羟基、羧基、-C(=O)OC1-6烷基、氨基、C1-6烷基、C2-6炔基、C1-6烷氧基、C3-12碳环、5至12元的杂环、-O-C3-12碳环、-NH-C3-12碳环或-CH2-C3-6环烷基,所述的羟基、氨基、烷基、炔基、烷氧基、碳环、杂环或环烷基任选进一步被0至4个选自F、Cl、Br、I、氰基、羟基、C1-6烷基、C2-6炔基、C1-4烷氧基、C3-6环烷基或C3-10碳环或5至10元的杂环的取代基所取代,所述的杂环含有1至4个选自N、O或S的杂原子;
R1a优选F、Cl、Br、-(=O)、硝基、氰基、羟基、氨基、羧基、-C(=O)OC1-4烷基、C1-4烷基、C2-4炔基、C1-4烷氧基、C3-10碳环、5至10元的杂环、-O-C3-10碳环、-NH-C3-10碳环或-CH2-C3-6环烷基,所述的羟基、氨基、烷基、炔基、烷氧基、碳环、杂环或环烷基任选进一步被0至4个选自F、Cl、Br、氰基、羟基、C1-4烷基、C1-4烷氧基、C3-6环烷基、C3-10碳环或5至10元的杂环的取代基所取代,所述的杂环含有1至4个选自N、O或S的杂原子;
R2、R3、R4、R5各自独立的选自C1-6亚烷基,优选C1-4亚烷基,所述的亚烷基任选进一步被0至4个选自F、Cl、Br、I、氰基、羟基、C1-6烷基、C1-6烷氧基、苯基或苯基-C1-4亚烷基的取代基所取代;
R2、R5更优选亚甲基、亚乙基、亚丙基、-CH2CH(CH3)-、-CH(CH3)CH2-、-C(CH3)2CH2-、-CH2C(CH3)2-、亚丁基、-CH(CH3)CH2CH2-、-CH2CH(CH3)CH2-或-CH2CH(CH3)CH2-;
R3、R4更优选亚甲基、亚乙基、亚丙基或亚丁基;
A选自C3-12碳环或5至12元杂环,优选C6-10碳环或5至10元杂环,所述的碳环或杂环任选进一步被0至5个F、Cl、Br、I、氰基、羟基、氨基、C1-6烷基、C2-6炔基、C1-4烷氧基或C3-6环烷基的取代基取代,且所述的杂环含有1至3个选自N、O或S的杂原子;
X1和X2各自独立的选自键、-O-、-C(=O)-、-C(=O)O-、-OC(=O)-、-S-、-S(=O)-、-S(=O)2-、-C(=O)NRx-、-NRxC(=O)-、-OC(=O)NRx-、-NRxC(=O)O-、-NRxC(=O)NRx、-NRxS(=O)2-、-S(=O)2NRx-、-NRxS(=O)2NRx或-NRx-;
Rx各自独立选自H、C1-6烷基或C3-8环烷基,所述的烷基或环烷基任选进一步被0至5个选自F、Cl、Br、I、C1-4烷基、C3-6环烷基或C1-4烷氧基的取代基所取代;
Rx优选H、C1-6烷基或C3-8环烷基,所述的烷基或环烷基任选进一步被0至5个选自F、Cl、Br、I、甲基、乙基、环丙基、环丁基、环戊基、环己基、甲氧基或乙氧基的取代基所取代;
Rx更优选H、甲基、乙基、丙基、异丙基、丁基、异丁基、戊基、环丙基、环丁基、环戊基、环己基、-CH2-环丙基、-CH2-环丁基或-CH2-环戊基;
离去基团选自氯、溴、碘、-OMs或OTs;
n1选自1、2或3;
本发明一种制备式(I)所示化合物的中间体的优选方案,该中间体选自式(M-1)、式(M-2)或式(M-3)所示化合物或者其立体异构体和药学上可接受的盐,
Rm1选自氨基保护基或H,优选叔丁基氧基羰基或H;
Rm2选自-COOH、-COOC1-6烷基、-X2-R5-OH、-X2-R5-离去基团、-X2-Rm3CHO、-X2-Rm3C(OC1-6烷基)2或所述烷基任选进一步被0至4个F、Cl、Br或C1-4烷基所取代,Rm2优选-COOH、-COOt-Bu或-X2-Rm3C(OCH3)2;
Rm3选自较R5少一个碳原子的亚烷基,Rm3优选亚甲基或亚乙基;
X1选自-O-;
X2选自键或者-C(=O)NRx-;
Rx选自H、甲基、乙基、丙基、异丙基、丁基、异丁基、戊基、环丙基、环丁基、环戊基、环己基、-CH2-环丙基、-CH2-环丁基或-CH2-环戊基;
R2、R3、R4、R5各自独立的选自亚甲基、亚乙基或亚丙基;
离去基团选自氯、溴、碘、-OMs或OTs;
n1选自1、2或3。
本发明一种制备式(I)所示化合物的中间体的优选方案,该中间体如下结构所示的化合物或者或者其立体异构体和药学上可接受的盐:
本发明优选化合物的制备方法任选通过如下途径之一制备(下述反应步骤部分可通过“一锅法”制备得到):
Rx选自H、甲基、乙基、丙基、异丙基、丁基、异丁基、戊基、环丙基、环丁基、环戊基、环己基、-CH2-环丙基、-CH2-环丁基或-CH2-环戊基;
Ra选自H、F、Cl、Br、甲基、乙基、异丙基或CF3;
化合物A(或其盐,优选三氟乙酸盐)与化合物B在碱性条件下发生取代反应得到化合物C,优选条件为将反应底物溶于二氧六环、乙腈、四氢呋喃、2-甲基四氢呋喃中,在1~6当量的叔丁醇钠、磷酸钾、
碳酸铯或碳酸钾存在下,可加入少量的水(为反应溶剂的3%~5%(v/v)),室温~回流反应。
化合物C水解生成化合物D(或其盐,优选三氟乙酸盐),优选条件为将反应底物溶于二氯甲烷中,加入三氟乙酸(为二氯甲烷体积的1/5~1/1),室温反应。
化合物E与化合物D(或其盐,优选三氟乙酸盐)发生取代反应得到化合物F,优选条件为将反应底物溶于二氯甲烷中,在1~6当量的三乙胺、4-二甲氨基吡啶或N,N-二异丙基乙胺存在下,加入1.0~3当量的HATU,室温反应。
化合物F在脱醛基保护条件下反应得到化合物G,优选条件为将反应底物溶于四氢呋喃中,加入1~10当量的乙酸或对甲苯磺酸一水合物,室温~50℃反应。
化合物G与化合物H1发生还原胺化反应得到化合物T1(本发明的优选化合物),优选条件为将反应底物溶于N-甲基吡咯烷酮或N,N-二甲基甲酰胺中,加入0.8~2当量的乙酸,室温搅拌0.5~1小时后,加入2~3当量的三乙酰氧基硼氢化钠,室温反应。
化合物G与化合物H2发生还原胺化反应得到化合物I,优选条件为将反应底物溶于二氯甲烷和甲醇的混合溶剂中(优选地v/v=10/3~10/1),搅拌0.5~1小时后,加入2~3当量的三乙酰氧基硼氢化钠,室温反应。
化合物I发生脱TBS保护得到化合物T2(本发明的优选化合物),优选条件为将反应底物溶于四氢呋喃中,加入5~20当量的三乙胺三氢氟酸,室温反应。
化合物C也可通过如下途径反应:
化合物A(或其盐,优选三氟乙酸盐)与化合物J在碱性条件下发生取代反应得到化合物K,优选条件为将反应底物溶于二氧六环、乙腈、四氢呋喃、2-甲基四氢呋喃中,在1~6当量的叔丁醇钠、磷酸钾、碳酸铯或碳酸钾存在下,可加入少量的水(为反应溶剂的3%~5%(v/v)),室温~回流反应。
作为选择,化合物A与羟基被保护了的化合物J在碱性条件下发生取代反应,然后在脱羟基保护条件下反应得到化合物K,其中,取代反应的优选条件为将反应底物溶于二氧六环、乙腈、四氢呋喃、2-甲基四氢呋喃中,在1~6当量的叔丁醇钠、磷酸钾、碳酸铯或碳酸钾存在下,可加入少量的水(为反应溶剂的3%~5%(v/v)),室温~回流反应。当羟基保护基为TBS时,将底物溶于四氢呋喃中,加入TBAF(四丁基氟化铵),室温反应。
化合物K与丙烯酸叔丁酯反应生成化合物C,优选条件为将反应底物溶于乙腈中,加入苄基三甲基氢氧化铵,室温反应。
化合物K也可通过如下途径反应得到:
化合物A(或其盐,优选三氟乙酸盐)与化合物N发生还原胺化反应得到化合物K,优选条件为将反应底物溶于四氢呋喃中,室温搅拌0.5~1小时后,加入2~3当量的三乙酰氧基硼氢化钠,室温反应。作为选择,化合物A与羟基被保护的化合物N发生还原胺化反应,然后在脱保护条件下反应生成化合物K。当羟基保护基为TBS时,将底物溶于四氢呋喃中,加入TBAF(四丁基氟化铵),室温反应。
化合物C也可通过如下途径反应得到:
化合物A(或其盐,优选三氟乙酸盐)与化合物M发生还原胺化反应得到化合物C,优选条件为将反应底物溶于四氢呋喃中,室温搅拌0.5~1小时后,加入2~3当量的三乙酰氧基硼氢化钠,室温反应。
化合物A(或其盐,优选三氟乙酸盐)可通过途径反应得到:
LG选自Cl、Br或I;
R1与上述合成途径一致;
PG选自叔丁基氧基羰基;
化合物A2与化合物A4在碱性条件下发生偶联反应生成化合物A1(W=键),优选条件为将反应底物溶于二氧六环、甲苯、二甲基亚砜或N,N-二甲基甲酰胺中,在1~3当量的叔丁醇钠、磷酸钾、碳酸铯或碳酸钾存在下,加入0~0.2当量的醋酸钯或三(二亚苄基丙酮)二钯催化剂和任选自0~0.2当量4,5-双二苯基膦-9,9-二甲基氧杂蒽、三苯基膦或三叔丁基膦的配体,在60~150℃范围内反应;
或者化合物A2与化合物A4在在碱性条件下发生取代反应生成化合物A1(W=键),优选条件为将反应底物溶于乙腈、二氧六环、甲苯、二甲基亚砜或N,N-二甲基甲酰胺中,在0.8~3当量的1,5-二氮杂二环[5.4.0]十一-5-烯、碳酸钾、碳酸钠、三乙胺、碳酸铯或碳酸钾存在下,室温~150℃反应。
化合物A3与化合物A4发生缩合反应生成化合物A1(W=-(C=O)),优选条件为将反应底物溶于二氯甲烷中,加入2~5当量的三乙胺和1~2当量的活性剂(优选1-丙基磷酸酐(T3P,CAS:68957-94-8)、HATU(CAS:148893-10-1或羰基二咪唑),室温反应。
除非有相反的陈述,在说明书和权利要求书中使用的术语具有下述含义。
本发明所述基团和化合物中所涉及的碳、氢、氧、硫、氮或卤素均包括它们的同位素,及本发明所述基团和化合物中所涉及的碳、氢、氧、硫、氮或卤素任选进一步被一个或多个它们对应的同位素所替代,其中碳的同位素包括12C、13C和14C,氢的同位素包括氕(H)、氘(D,又称为重氢)、氚(T,又称为超重氢),氧的同位素包括16O、17O和18O,硫的同位素包括32S、33S、34S和36S,氮的同位素包括14N和15N,氟的同位素19F,氯的同位素包括35Cl和37Cl,溴的同位素包括79Br和81Br。
“烷基”是指直链和支链的一价饱和烃基,主链包括1至10个碳原子,优选为1至8个碳原子,进一步优选为1至6个碳原子,更优选为1至4个碳原子的直链与支链基团,最优选1至2个碳原子,烷基的实例包括但不限于甲基、乙基、正丙基、异丙基、正丁基、异丁基、仲丁基、叔丁基、正戊基、2-戊基、3-戊基、2-甲基-2-丁基、3-甲基-2-丁基、正己基、正庚基、正辛基、正壬基和正癸基等;所述的烷基可以任选进一步被0至5个选自F、Cl、Br、I、=O、-CH2F、-CHF2、-CF3、-OCH2F、-OCHF2、-OCF3、羟基、-SR19、硝基、氰基、异氰基、烷基、羟基烷基、烷氧基、碳环基、杂环基、C2-8烯基、C2-8炔基、-(CH2)k-C(=O)-R19、-(CH2)k-C(=O)-O-R19、-(CH2)k-C(=O)-NR19R19a、-(CH2)k-S(=O)j-R19、-O-C(=O)-O-R19或者-NR19R19a的取代基所取代,其中R19和R19a各自独立选自H、羟基、氨基、羧基、C1-8烷基、C1-8烷氧基、C2-8烯基、C2-8炔基、3至10元碳环基、4至10元杂环基、3至10元碳环基氧基或者4至10元杂环基氧基,k选自0、1、2、3、4或者5,j选自0、1或者2。本文中出现的烷基、k、j或R18和R18a,其定义如上所述。
“亚烷基”是指直链和支链的二价饱和烃基,包括-(CH2)v-(v为1至10的整数),亚烷基实施例包括但不限于亚甲基、亚乙基、亚丙基和亚丁基等;所述的亚烷基可以任选进一步被0至5个选自F、Cl、Br、I、=O、-CH2F、-CHF2、-CF3、-OCH2F、-OCHF2、-OCF3、羟基、-SR19、硝基、氰基、异氰基、烷基、羟基烷基、烷氧基、碳环基、杂环基、C2-8烯基、C2-8炔基、-(CH2)k-C(=O)-R19、-(CH2)k-C(=O)-O-R19、-(CH2)k-C(=O)-NR19R19a、-(CH2)k-S(=O)j-R19、-O-C(=O)-O-R19或者-NR19R19a的取代基所取代,当亚烷基中的取代基数量大于等于2个时,取代基可以稠合在一起形成环状结构。本文中出现的亚烷基,其定义如上所述。
“烷氧基”是指O-烷基的一价基团,其中,烷基如本文所定义,烷氧基实施例包括但不限于甲氧基、乙氧基、1-丙氧基、2-丙氧基、1-丁氧基、2-甲基-1-丙氧基、2-丁氧基、2-甲基-2-丙氧基、1-戊氧基、2-戊氧基、3-戊氧基、2-甲基-2-丁氧基、3-甲基-2-丁氧基、3-甲基-1-丁氧基和2-甲基-1-丁氧基等。
“烯基”是指直链和支链的一价不饱和烃基,其具有至少1个,通常有1、2或3个碳碳双键,主链包括2至10个碳原子,进一步优选2至6个碳原子,更优选在主链上有2至4个碳原子,烯基实施例包括但不限于乙烯基、烯丙基、1-丙烯基、2-丙烯基、1-丁烯基、2-丁烯基、3-丁烯基、1-戊烯基、2-戊烯基、3-戊烯基、4-戊烯基、1-甲基-1-丁烯基、2-甲基-1-丁烯基、2-甲基-3-丁烯基、1-己烯基、2-己烯基、3-己烯基、4-己烯基、5-己烯基、1-甲基-1-戊烯基、2-甲基-1-戊烯基、1-庚烯基、2-庚烯基、3-庚烯基、4-庚烯基、1-辛烯基、3-辛烯基、1-壬烯基、3-壬烯基、1-癸烯基、4-癸烯基、1,3-丁二烯、1,3-戊二烯、1,4-戊二烯和1,4-己二烯等;所述的烯基可以任选进一步被0至5个选自F、Cl、Br、I、=O、-CH2F、-CHF2、-CF3、-OCH2F、-OCHF2、-OCF3、羟基、-SR19、硝基、氰基、异氰基、烷基、羟基烷基、烷氧基、碳环
基、杂环基、C2-8烯基、C2-8炔基、-(CH2)k-C(=O)-R19、-(CH2)k-C(=O)-O-R19、-(CH2)k-C(=O)-NR19R19a、-(CH2)k-S(=O)j-R19、-O-C(=O)-O-R19或者-NR19R19a的取代基所取代。本文中出现的烯基,其定义如上所述。
“炔基”是指直链和支链的一价不饱和烃基,其具有至少1个,通常有1、2或3个碳碳三键,主链包括2至10个碳原子,进一步优选2至6个碳原子,更优选在主链上有2至4个碳原子,炔基实施例包括但不限于乙炔基、1-丙炔基、2-丙炔基、丁炔基、2-丁炔基、3-丁炔基、1-甲基-2-丙炔基、4-戊炔基、3-戊炔基、1-甲基-2-丁炔基、2-己炔基、3-己炔基、2-庚炔基、3-庚炔基、4-庚炔基、3-辛炔基、3-壬炔基和4-癸炔基等;所述的炔基可以任选进一步被0至5个选自F、Cl、Br、I、=O、-CH2F、-CHF2、-CF3、-OCH2F、-OCHF2、-OCF3、羟基、-SR19、硝基、氰基、异氰基、烷基、羟基烷基、烷氧基、碳环基、杂环基、C2-8烯基、C2-8炔基、-(CH2)k-C(=O)-R19、-(CH2)k-C(=O)-O-R19、-(CH2)k-C(=O)-NR19R19a、-(CH2)k-S(=O)j-R19、-O-C(=O)-O-R19或者-NR19R19a的取代基所取代。本文中出现的炔基,其定义如上所述。
“环烷基”是指一价饱和的碳环烃基,通常有3至10个碳原子,非限制性实施例包括环丙基、环丁基、环戊基、环己基或环庚基等。所述的环烷基可以任选进一步被0至5个选自F、Cl、Br、I、=O、-CH2F、-CHF2、-CF3、-OCH2F、-OCHF2、-OCF3、羟基、-SR19、硝基、氰基、异氰基、烷基、羟基烷基、烷氧基、碳环基、杂环基、C2-8烯基、C2-8炔基、-(CH2)k-C(=O)-R19、-(CH2)k-C(=O)-O-R19、-(CH2)k-C(=O)-NR19R19a、-(CH2)k-S(=O)j-R19、-O-C(=O)-O-R19或者-NR19R19a的取代基所取代。本文中出现的环烷基,其定义如上所述。
“碳环”或“碳环基”是指饱和或者不饱和的芳香环或者非芳香环,芳香环或者非芳香环可以是3至10元的单环、4至12元双环或者10至15元三环体系,碳环基可以连接有桥环或者螺环,非限制性实施例包括环丙基、环丁基、环戊基、1-环戊基-1-烯基、1-环戊基-2-烯基、1-环戊基-3-烯基、环己基、1-环己基-2-烯基、1-环己基-3-烯基、环己烯基、环己二烯基、环庚基、环辛基、环壬基、环癸基、环十一烷基、环十二烷基、苯基、萘基、苯并环丙烯基、2,3-二氢苯并环丙烯基、所述的碳环基可以任选进一步被0至5个选自F、Cl、Br、I、=O、-CH2F、-CHF2、-CF3、-OCH2F、-OCHF2、-OCF3、羟基、-SR19、硝基、氰基、异氰基、烷基、羟基烷基、烷氧基、碳环基、杂环基、C2-8烯基、C2-8炔基、-(CH2)k-C(=O)-R19、-(CH2)k-C(=O)-O-R19、-(CH2)k-C(=O)-NR19R19a、-(CH2)k-S(=O)j-R19、-O-C(=O)-O-R19或者-NR19R19a的取代基所取代。本文中出现的碳环基,其定义如上所述。
“杂环”或“杂环基”是指饱和或不饱和的芳香环或者非芳香环,芳香环或者非芳香环可以是3至10元的单环、4至12元双环或者10至15元三环体系,且包含1至4个选自N、O或S的杂原子,优选3至10元杂环基,杂环基的环中选择性取代的N、S可被氧化成各种氧化态。杂环基可以连接在杂原子或者碳原子上,杂环基可以连接有桥环或者螺环,非限制性实施例包括环氧乙基、环氧丙基、氮杂环丙基、氧杂环丁基、氮杂环丁基、硫杂环丁基、1,3-二氧戊环基、1,4-二氧戊环基、1,3-二氧六环基、氮杂环庚
基、氧杂环庚基、硫杂环庚基、氧氮杂卓基、二氮杂卓基、硫氮杂卓基、吡啶基、哌啶基、高哌啶基、呋喃基、噻吩基、吡喃基、N-烷基吡咯基、嘧啶基、吡嗪基、哒嗪基、哌嗪基、高哌嗪基、咪唑基、哌啶基、哌叮基、吗啉基、硫代吗啉基、噻噁烷基、1,3-二噻基、二氢呋喃基、二氢吡喃基、二噻戊环基、四氢呋喃基、四氢噻吩基、四氢吡喃基、四氢噻喃基、四氢吡咯基、四氢咪唑基、四氢噻唑基、四氢吡喃基、苯并咪唑基、苯并吡啶基、吡咯并吡啶基、苯并二氢呋喃基、2-吡咯啉基、3-吡咯啉基、二氢吲哚基、2H-吡喃基、4H-吡喃基、二氧杂环己基、1,3-二氧戊基、吡唑啉基、二噻烷基、二噻茂烷基、二氢噻吩基、吡唑烷基、咪唑啉基、咪唑烷基、1,2,3,4-四氢异喹啉基、1,2,3,4-四氢喹啉基、苯并三氮唑基、3-氮杂双环[3.1.0]己基、3-氮杂双环[4.1.0]庚基、氮杂双环[2.2.2]己基、3H-吲哚基喹嗪基、N-吡啶基尿素、1,1-二氧硫代吗啉基、氮杂二环[3.2.1]辛烷基、氮杂二环[5.2.0]壬烷基、氧杂三环[5.3.1.1]十二烷基、氮杂金刚烷基和氧杂螺[3.3]庚烷基。所述的杂环基可以任选进一步被0至5个选自F、Cl、Br、I、=O、-CH2F、-CHF2、-CF3、-OCH2F、-OCHF2、-OCF3、羟基、-SR19、硝基、氰基、异氰基、烷基、羟基烷基、烷氧基、碳环基、杂环基、C2-8烯基、C2-8炔基、-(CH2)k-C(=O)-R19、-(CH2)k-C(=O)-O-R19、-(CH2)k-C(=O)-NR19R19a、-(CH2)k-S(=O)j-R19、-O-C(=O)-O-R19或者-NR19R19a的取代基所取代。本文中出现的杂环基,其定义如上所述。
Bn为苄基,即-CH2苯基。
“β-肾上腺素受体结合基团”是指能够与β-肾上腺素能受体结合的基团;诸如参见综述文章“β-adrenergic receptors in Comprehensive Medicinal Chemistry,1990,B.E.Main,p187(Pergamon Press)”。上述基团也参见例如WO/2005092841、US/20050215542、WO/2005070872、WO/2006023460、WO/2006051373、WO/2006087315和WO/2006032627。非限制性实施例包括R6、R7各自独立的选自H或C1-4烷基,R8选自H或OH,B选自
其中Q选自-CH=CH-、-CH2CH2-、-O-、-S-、-CH2O-、-OCH2-、-C(CH3)2O-或-OC(CH3)2-。
“任选”或“任选地”是指随后所描述的事件或环境可以但不必须发生,该说明包括该事件或环境发生或不发生的场合。如:“任选被F取代的烷基”指烷基可以但不必须被F取代,说明包括烷基被F取代的情形和烷基不被F取代的情形。
“药物组合物”表示一种或多种文本所述化合物或其生理学/药学上可接受的盐与其他组成成分的混
合物,其中其它组分包含生理学/药学上可接受的载体和赋形剂。
“载体”指的是不会对生物体产生明显刺激且不会消除所给予化合物的生物活性和特性的载体或稀释剂。
“赋形剂”指的是加入到药物组合物中以进一步依赖于化合物给药的惰性物质。赋形剂的实例包括但不限于碳酸钙、磷酸钙、各种糖和不同类型的淀粉、纤维素衍生物(包括微晶纤维素)、明胶、植物油、聚乙二醇类、稀释剂、成粒剂、润滑剂、粘合剂、崩解剂等。
“前药”是指可以在生理条件下或通过溶剂解转化为具有生物活性的本发明化合物的化合物。本发明的前药通过修饰本发明化合物中的功能基团来制备,该修饰可以通过常规的操作或者在体内被除去,而得到母体化合物。
“立体异构体”是指由分子中原子在空间上排列方式不同所产生的异构体,包括顺反异构体、对映异构体和构象异构体。
“有效剂量”指引起组织、系统或受试者生理或医学翻译的化合物的量,此量是所寻求的,包括在受治疗者身上施用时足以预防受治疗的疾患或病症的一种或几种症状发生或使其减轻至某种程度的化合物的量。
“溶剂化物”指本发明化合物或其盐,它们还包括以分子间非共价力结合的化学计量或非化学计量的溶剂。当溶剂为水时,则为水合物。
“IC50”指半数抑制浓度,指达到最大抑制效果一半时的浓度。
以下通过具体实施例详细说明本发明的实施过程和产生的有益效果,旨在帮助阅读者更好地理解本发明的实质和特点,不作为对本案可实施范围的限定。
化合物的结构是通过核磁共振(NMR)或(和)质谱(MS)来确定的。NMR位移(δ)以10-6(ppm)的单位给出。NMR的测定是用(Bruker Avance III 400和Bruker Avance 300)核磁仪,测定溶剂为氘代二甲基亚砜(DMSO-d6),氘代氯仿(CDCl3),氘代甲醇(CD3OD),内标为四甲基硅烷(TMS)。MS的测定用(Agilent 6120B(ESI)和Agilent 6120B(APCI))。HPLC的测定使用安捷伦1260DAD高压液相色谱仪(Zorbax SB-C18 100×4.6mm)。薄层层析硅胶板使用烟台黄海HSGF254或青岛GF254硅胶板,薄层色谱法(TLC)使用的硅胶板采用的规格是0.15mm~0.20mm,薄层层析分离纯化产品采用的规格是0.4mm~0.5mm。柱层析一般使用烟台黄海硅胶200~300目硅胶为载体。本发明的已知的起始原料可以采用或按照本领域已知的方法来合成,或可购买于泰坦科技、安耐吉化学、上海德默、成都科龙化工、韶远化学科技、百灵威科技等公司。氮气氛是指反应瓶连接一个约1L容积的氮气气球。氢气氛是指反应瓶连接一个约1L容积的氢气气球。氢化反应通常抽真空,充入氢气,反复操作3次。实施例中无特殊说明,反应在氮气氛下进行。实施例中无特殊说明,溶液是指水溶液。实施例中无特殊说明,反应的温度为室温。室温为最适宜的反应温度,为20℃~30℃。实施例中无特殊说明,M为mol/L。TBS为叔丁基二甲基硅基。Boc为叔丁基氧基羰基。Bn为苄基。TsOH·H2O为对甲苯磺酸一水合物。DMF为N,N-二甲基甲酰胺。THF为四氢呋喃。DCM为二氯甲烷。
HATU:(2-(7-氧化苯并三氮唑)-N,N,N',N'-四甲基脲六氟磷酸酯,CAS:148893-10-1)。
中间体1:3-[2-[3-(2-溴乙基)苯基]乙氧基]丙酸叔丁酯
tert-butyl 3-[2-[3-(2-bromoethyl)phenyl]ethoxy]propanoate
第一步:2-(3-(2-((叔丁基二甲基硅基)氧)乙基)苯基)乙醇(1b)
2-(3-(2-((tert-butyldimethylsilyl)oxy)ethyl)phenyl)ethan-1-ol
将钠氢(4.3g,107.0mmol,60%w/w)加入THF(100mL)中,0℃滴加2,2’-(1,3-亚苯基)二乙醇(1a)(18.0g,108.0mmol)的THF溶液(150mL),滴加完毕后,0℃搅拌30分钟,滴加叔丁基二甲基氯硅烷(14.5g,108.0mmol),室温搅拌10小时。用水淬灭反应(150mL),减压浓缩除去大部分THF,用乙酸乙酯(200mL×2)萃取,合并后的有机相用饱和食盐水(100mL×1)洗涤,有机相用无水硫酸钠干燥,过滤,滤液减压浓缩后,残留物用硅胶柱层析分离(石油醚/乙酸乙酯(v/v)=4:1)得到黄色液体状的1b(10.2g,产率34%)。
1H NMR(400MHz,CDCl3)δ7.27-7.22(m,1H),7.10-7.07(m,3H),3.87-3.80(m,4H),2.87-2.80(m,4H),0.89(s,9H),0.00(s,6H).
第二步:3-[2-[3-[2-[叔丁基(二甲基)硅基]氧乙基]苯基]乙氧基]丙酸叔丁酯(1c)
tert-butyl 3-[2-[3-[2-[tert-butyl(dimethyl)silyl]oxyethyl]phenyl]ethoxy]propanoate
将1b(10.2g,36.4mmol)溶于乙腈(30mL)中,加入丙烯酸叔丁酯(14.0g,109.0mmol)和苄基三甲基氢氧化铵(4.6g,10.9mmol,40%的甲醇溶液),室温搅拌3小时。反应液直接减压浓缩除去大部分反应溶剂,残留物用硅胶柱层析分离(石油醚/乙酸乙酯(v/v)=10:1)得到黄色液体状的1c(12.0g,产率81%)。
1H NMR(400MHz,CDCl3)δ7.20-7.16(m,1H),7.06-7.02(m,3H),3.81-3.77(m,2H),3.70-3.60(m,4H),2.84-2.77(m,4H),2.50-2.46(m,2H),1.44(s,9H),0.87(s,9H),-0.02(s,6H).
第三步:3-[2-[3-[2-羟基乙基]苯基]乙氧基]丙酸叔丁酯(1d)
tert-butyl 3-[2-[3-(2-hydroxyethyl)phenyl]ethoxy]propanoate
将1c(12.0g,29.4mmol)溶于THF(100mL)中,加入四丁基氟化铵(15.4g,58.7mmol),室温搅拌3小时。反应液直接减压浓缩除去大部分反应溶剂,加入水(100mL)用乙酸乙酯(200mL×2)萃取,合
并后的有机相用无水硫酸钠干燥,过滤,滤液减压浓缩后,残留物用柱层析分离(石油醚/乙酸乙酯(v/v)=3:1)得到黄色固体状的1d(7.2g,产率83%)。
1H NMR(400MHz,CDCl3)δ7.24-7.20(m,1H),7.09-7.05(m,3H),3.86-3.83(m,2H),3.70-3.63(m,4H),2.87-2.82(m,4H),2.49-2.46(m,2H),1.43(s,9H).
第四步:3-[2-[3-(2-溴乙基)苯基]乙氧基]丙酸叔丁酯(中间体1)
tert-butyl 3-[2-[3-(2-bromoethyl)phenyl]ethoxy]propanoate
将1d(7.2g,24.5mmol)溶于二氯甲烷(100mL)中加入咪唑(3.3g,49.0mmol),三苯基膦(9.6g,37.0mmol),四溴化碳(12.0g,37.0mmol),搅拌2小时。加入饱和碳酸氢钠水溶液(100mL)淬灭反应,萃取,水相用二氯甲烷(150mL×1)萃取,合并后的有机相用无水硫酸钠干燥,过滤,滤液减压浓缩后柱层析分离(石油醚/乙酸乙酯(v/v)=10:1)得到黄色固体状的中间体1(7.5g,产率86%)。
1H NMR(400MHz,CDCl3)δ7.23-7.21(m,1H),7.12-7.10(m,1H),7.06-7.04(m,2H),3.70-3.63(m,4H),3.58-3.54(m,2H),3.13-3.12(m,2H),2.87-2.82(m,2H),2.50-2.46(m,2H),1.44(s,9H。
中间体2:3-[2-[3-(溴甲基)苯基]乙氧基]丙酸叔丁酯(中间体2)
tert-butyl 3-[2-[3-(bromomethyl)phenyl]ethoxy]propanoate
第一步:3-[2-(3-溴苯基)乙氧基]丙酸叔丁酯(2b)
tert-butyl 3-[2-(3-bromophenyl)ethoxy]propanoate
将间溴苯乙醇(2a)(80.4g,0.4mol)置于乙腈(200mL)中,加入丙烯酸叔丁酯(76.9g,0.6mol),后滴加苄基三甲基氢氧化铵(50.2g,0.12mol,40%的甲醇溶液),40℃搅拌5小时。减压浓缩除去大部分反应溶剂,加入水(200mL)和二乙胺(50mL),室温搅拌30分钟。向反应液中加入乙酸乙酯(500mL)萃取,水相用乙酸乙酯(500mL×1)萃取,合并后的有机相用饱和食盐水洗(200mL),无水硫酸钠干燥,过滤,滤液减压浓缩后,残留物用硅胶柱层析分离(石油醚/乙酸乙酯(v/v)=12:1)得到黄色液体状的2b(94.0g,产率71.4%)。
1H NMR(400MHz,CDCl3)δ7.36-7.29(m,2H),7.15-7.10(m,2H),3.68-3.61(m,4H),2.84-2.81(m,2H),2.48-2.45(t,3H),1.43(s,9H).
第二步:3-[2-(3-氰基苯基)乙氧基]丙酸叔丁酯(2c)
tert-butyl 3-[2-(3-cyanophenyl)ethoxy]propanoate
将2b(9.9g,30.0mmol)溶于DMF(60mL)中,加入氰化亚铜(5.4g,60.0mmol),150℃搅拌16小时。反应冷却至室温,加入水(100mL)和乙酸乙酯(100mL),硅藻土过滤,滤饼依次用水(100mL×1)和乙酸乙酯(100mL×1)洗涤,萃取滤液,水相用乙酸乙酯(200mL×1)萃取,合并后的有机相用饱和食盐水(100mL×1)洗涤,无水硫酸钠干燥,过滤,滤液减压浓缩后,残留物用硅胶柱层析分离(石油醚/乙
酸乙酯(v/v)=10:1)得到黄色液体状的2c(7.2g,产率87%)。
1H NMR(400MHz,CDCl3)δ7.50-7.44(m,3H),7.37-7.33(m,1H),3.67-3.63(m,4H),2.89-2.85(t,3H),2.46-2.42(m,2H),1.41(s,9H).
LCMS m/z=298.1[M+23]。
第三步:3-[2-(3-甲酰基苯基)乙氧基]丙酸叔丁酯(2d)
tert-butyl 3-[2-(3-formylphenyl)ethoxy]propanoate
将2c(35.2g,128mmol)中加入吡啶(120mL)、乙酸(80mL)和水(80mL),加入雷尼镍(1.8g,30mmol)和次亚磷酸钠(27.1g,256mmol),45℃搅拌3小时。硅藻土过滤,滤饼依次用水(200mL)和乙酸乙酯(300mL)洗涤,减压浓缩除去大部分反应溶剂,加入水(100mL)用乙酸乙酯(200mL×2)萃取,合并后的有机相用无水硫酸钠干燥,过滤,滤液减压浓缩后,残留物用硅胶柱层析分离(石油醚/乙酸乙酯(v/v)=3:1)得到黄色液体状的2d(22.0g,产率61.8%)。
LCMS m/z=301.2[M+23]。
第四步:3-[2-[(3-羟基甲基)苯基]乙氧基]丙酸叔丁酯(2e)
tert-butyl 3-[2-[3-(hydroxymethyl)phenyl]ethoxy]propanoate
将2d(22.0g,79.1mmol)溶于甲醇(200mL)中,加入硼氢化钠(6.0g,158.2mmol),室温搅拌2小时。加入水(100mL),减压浓缩除去大部分甲醇,用乙酸乙酯(300mL×2)萃取,合并后的有机相用无水硫酸钠干燥,过滤,滤液减压浓缩后,残留物用硅胶柱层析分离(石油醚/乙酸乙酯(v/v)=2:1)得到黄色液体状的2e(18.0g,产率81.2%)。
1H NMR(400MHz,CDCl3)δ7.28-7.23(m,2H),7.19-7.17(m,1H),7.14-7.12(m,1H),4.65(s,2H),3.69-3.64(m,4H),2.89-2.85(t,2H),2.48-2.45(t,2H),1.43(s,9H).
第五步:3-[2-[3-(溴甲基)苯基]乙氧基]丙酸叔丁酯(中间体2)
tert-butyl 3-[2-[3-(bromomethyl)phenyl]ethoxy]propanoate
将2e(12.0g,42.8mmol)溶于DCM(200mL)中,0℃,加入三苯基膦(13.5g,51.4mmol)、四溴化碳(17.0g,51.4mmol)和咪唑(5.8g,85.6mmol),室温搅拌2小时。加入饱和碳酸氢钠水溶液(150mL),萃取,水相用DCM(200mL×1)萃取,合并后的有机相用无水硫酸钠干燥,过滤,滤液减压浓缩后,残留物用硅胶柱层析分离(石油醚/乙酸乙酯(v/v)=10:1)得到黄色液体状的中间体2(4.0g,产率27.0%)。
中间体3:3-[2-[4-(2-溴乙基)苯基]乙氧基]丙酸叔丁酯(中间体3)
tert-butyl 3-[2-[4-(2-bromoethyl)phenyl]ethoxy]propanoate
第一步:2-(4-(2-((叔丁基二甲基硅基)氧)乙基)苯基)乙醇(3b)
2-(4-(2-((tert-butyldimethylsilyl)oxy)ethyl)phenyl)ethan-1-ol
将钠氢(4.0g,100.0mmol,60%w/w)置于THF(100mL)中,0℃滴加2,2’-(1,3-亚苯基)二乙醇(3a)(16.6g,100.0mmol)的THF溶液(150mL),0℃搅拌30分钟,然后滴加叔丁基二甲基氯硅烷(15.0g,100.0mmol),室温搅拌10小时。滴加水淬灭反应(150mL),减压浓缩除去大部分THF,用乙酸乙酯萃取(200mL×2),合并后的有机相用饱和食盐水洗(100mL),无水硫酸钠干燥,过滤,滤液减压浓缩后,残留物用硅胶柱层析分离(石油醚/乙酸乙酯(v/v)=4:1)得到黄色液体状的3b(11.2g,产率40%)。
LCMS m/z=303.3[M+23]。
第二步:3-[2-[4-[2-[叔丁基(二甲基)硅基]氧乙基]苯基]乙氧基]丙酸叔丁酯(3c)
tert-butyl 3-[2-[4-[2-[tert-butyl(dimethyl)silyl]oxyethyl]phenyl]ethoxy]propanoate
将3b(11.2g,40.0mmol)溶于乙腈(30mL)中,加入丙烯酸叔丁酯(10.3g,80.0mmol)和苄基三甲基氢氧化铵(5.0g,12.0mmol,40%的甲醇溶液),室温搅拌3小时。减压浓缩除去大部分反应溶剂,残留物用硅胶柱层析分离(石油醚/乙酸乙酯(v/v)=10:1)得到黄色液体状的3c(13.9g,产率85%)。
第三步:3-[2-[4-[2-羟基乙基]苯基]乙氧基]丙酸叔丁酯(3d)
tert-butyl 3-[2-[4-(2-hydroxyethyl)phenyl]ethoxy]propanoate
将3c(13.9g,34.0mmol)溶于THF(100mL)中,加入四丁基氟化铵(17.8g,68.0mmol),室温搅拌3小时。减压浓缩除去大部分反应溶剂,加入水(100mL),用乙酸乙酯(200mL×2)萃取,合并后的有机相用无水硫酸钠干燥,过滤,滤液减压浓缩后,残留物用硅胶柱层析分离(石油醚/乙酸乙酯(v/v)=3:1)得到黄色固体状的3d(8.0g,产率80%)。
LCMS m/z=317.1[M+23]。
第四步:3-[2-[4-(2-溴乙基)苯基]乙氧基]丙酸叔丁酯(中间体3)
tert-butyl 3-[2-[4-(2-bromoethyl)phenyl]ethoxy]propanoate
将3d(8.0g,27.2mmol)溶于DCM(100mL)中,加入咪唑(3.7g,54.4mmol),三苯基膦(8.6g,32.6mmol),四溴化碳(10.8g,32.6mmol),室温搅拌2小时加入饱和碳酸氢钠水溶液(100mL),萃取,水相用DCM(150mL×1)萃取,合并后的有机相用无水硫酸钠干燥,过滤,滤液减压浓缩后,残留物用硅胶柱层析分离(石油醚/乙酸乙酯(v/v)=10:1)得到黄色液体状的中间体3(7.8g,产率80%)。
LCMS m/z=379.1[M+23]。
中间体4:3-[2-[4-(氯甲基)苯基]乙氧基]丙酸叔丁酯(中间体4)
tert-butyl 3-[2-[3-(chloromethyl)phenyl]ethoxy]propanoate
第一步:3-[2-(4-溴苯基)乙氧基]丙酸叔丁酯(4b)
tert-butyl 3-[2-(4-bromophenyl)ethoxy]propanoate
将4a(50.3g,0.25mol)置于乙腈(100mL)中,加入丙烯酸叔丁酯(48.1g,0.375mol),后滴加苄基三甲基氢氧化铵(31.4g,0.075mol,40%的甲醇溶液),40℃搅拌5小时。减压浓缩除去大部分反应溶剂,加入水(200mL)和二乙胺(50mL),室温搅拌30分钟。反应液用乙酸乙酯(500mL×2)萃取,合并后的有机相用饱和食盐水(200mL×1)洗涤,无水硫酸钠干燥,过滤,滤液减压浓缩后,残留物用硅胶柱层析分离(石油醚/乙酸乙酯(v/v)=12:1)得到黄色液体状的4b(61.7g,产率75%)。
第二步:3-[2-(4-氰基苯基)乙氧基]丙酸叔丁酯(4c)
tert-butyl 3-[2-(4-cyanophenyl)ethoxy]propanoate
将4b(9.9g,30.0mmol)溶于N,N-二甲基甲酰胺(60mL)中,加入氰化亚铜(5.4g,60.0mmol),150℃搅拌16小时。反应冷却至室温,加入水(100mL)和乙酸乙酯(100mL),硅藻土过滤,滤饼依次用水(100mL)和乙酸乙酯(100mL)洗涤,萃取滤液,水相用乙酸乙酯(200mL×1)萃取,合并后的有机相用饱和食盐水(100mL×1)洗涤,无水硫酸钠干燥,过滤,滤液减压浓缩后,残留物用硅胶柱层析分离(石油醚/乙酸乙酯(v/v)=10:1)得到黄色液体状的4c(7.2g,产率87%)。
LCMS m/z=298.1[M+23]。
第三步:3-[2-(4-甲酰基苯基)乙氧基]丙酸叔丁酯(4d)
tert-butyl 3-[2-(4-formylphenyl)ethoxy]propanoate
将4c(35.2g,128mmol)加入吡啶(120mL)、乙酸(80mL)和水(80mL)的混合物中,加入雷尼镍(1.8g,30mmol),次亚磷酸钠(27.1g,256mmol),45℃搅拌3小时。硅藻土过滤,滤饼依次用水(200mL)和乙酸乙酯(300mL)洗涤,滤液减压浓缩除去大部分反应溶剂,向残余物中加入水(100mL)和乙酸乙酯(200mL),萃取,水相用乙酸乙酯(200mL×1)萃取,合并后的有机相用无水硫酸钠干燥,过滤,滤液减压浓缩后,残留物用硅胶柱层析分离(石油醚/乙酸乙酯(v/v)=3:1)得到黄色液体状的4d(22.0g,产率61.8%)。
第四步:3-[2-[(4-羟基甲基)苯基]乙氧基]丙酸叔丁酯(4e)
tert-butyl 3-[2-[4-(hydroxymethyl)phenyl]ethoxy]propanoate
将4d(22.0g,79.1mmol)溶于甲醇(200mL)中,加入硼氢化钠(6.0g,158.2mmol),搅拌2小时。加入水(100mL),减压浓缩除去大部分甲醇,用乙酸乙酯(300mL×2)萃取,合并后的有机相用无水硫酸钠干燥,过滤,滤液减压浓缩后,残留物用硅胶柱层析分离(石油醚/乙酸乙酯(v/v)=2:1)得到黄色液体状的4e(18.0g,产率81.2%)。
1H NMR(400MHz,CDCl3)δ7.33-7.28(m,2H),7.21-7.19(m,2H),4.64(s,2H),3.67-3.64(m,4H),2.89-2.85(t,2H),2.48-2.45(t,2H),1.43(s,9H).
第五步:3-[2-[4-(氯甲基)苯基]乙氧基]丙酸叔丁酯(中间体4)
tert-butyl 3-[2-[4-(chloromethyl)phenyl]ethoxy]propanoate
将4e(11.2g,40.0mmol)溶于DCM(150mL)中,0℃加入三乙胺(10.1g,100.0mmol)和4-二甲胺基吡啶(0.24g,2.0mmol),滴加甲烷磺酰氯(6.9g,60.0mmol),室温搅拌48小时。加入饱和碳酸氢钠水溶液(150mL),萃取,水相用DCM(200mL×1)萃取,合并后的有机相用无水硫酸钠干燥,过滤,滤液减压浓缩后,残留物用硅胶柱层析分离(石油醚/乙酸乙酯(v/v)=10:1)得到黄色液体状的中间体4(7.2g,产率
60.0%)。
LCMS m/z=321.0[M+23]。
中间体5:N-(2,2-二甲氧基乙基)丁基-1-胺(中间体5)
N-(2,2-dimethoxyethyl)butan-1-amine
将正丁胺(5a)(14.6g,0.2mol)加入甲醇(150mL)中,加入乙二醛-1,1-二甲基乙缩醛水溶液(5b)(43.2g,0.22mol,60%水溶液),室温搅拌6小时后,分批加入硼氢化钠(11.3g,0.3mol),室温反应2小时。减压浓缩后除去大部分反应溶剂,加入水(200mL),用DCM萃取(300mL×2),合并后得有机相用无水硫酸钠干燥,减压浓缩后得到黄色液体状的中间体5(30.0g,产率93%)。
1H NMR(400MHz,CDCl3)δ4.45-4.42(m,1H),3.35(s,6H),2.70-2.69(m,2H),2.60-2.56(m,2H),1.44-1.40(m,2H),1.33-1.26(m,2H),0.90-0.86(m,3H)。
LCMS m/z=162.2[M+1]。
中间体6:3-[2-[3-(氯甲基)苯基]乙氧基]丙酸叔丁酯
tert-butyl 3-[2-[3-(chloromethyl)phenyl]ethoxy]propanoate
将2e(91.0g,324.5mmol)加入DMF(450mL)中,加入N,N-二甲基吡啶-4-胺(3.97g,32.5mmol)和三乙胺(110mL,811.5mmol),0℃搅拌5分钟后,缓慢滴入甲磺酰氯(37.7mL,486.8mmol),室温反应8小时。加入饱和碳酸氢钠水溶液(100mL),用DCM(200mL×3)萃取,合并后的有机相用无水硫酸钠干燥,减压浓缩后柱层析分离(石油醚:乙酸乙酯(v/v)=1:0~8:1)得到黄色液体状的中间体6(54.5g,产率56%)。
LCMS m/z=321.1[M+23].
中间体7:3-[2-[3-[[4-(4-乙基-2-吡啶基)-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-基]甲基]-2-氟苯基]乙氧基]丙烯酸叔丁酯
tert-butyl 3-[2-[3-[[4-(4-ethyl-2-pyridyl)-1-oxa-4,9-diazaspiro[5.5]undecan-9-yl]methyl]-2-fluoro-phenyl]ethoxy]propanoate
第一步:叔丁基[2-(2-氟苯基)乙氧基]二甲基硅烷(7b)
tert-butyl-[2-(2-fluorophenyl)ethoxy]-dimethyl-silane
将7a(21.03g,150.0mmol)加入DMF(150mL),0℃加入咪唑(30.64g,450.1mmol)和叔丁基二甲基氯硅烷(24.88g,165.0mmol),10min后升至室温反应3.5小时。加入水(200mL),用乙酸乙酯(200mL×2)萃取,合并有机相,饱和食盐水(40mL)洗,无水硫酸钠干燥,过滤,滤液减压浓缩,得到无色液体状的7b(35.2g,产率:92%)。
第二步:3-[2-[叔丁基(二甲基)硅基]氧乙基]-2-氟苯甲醛(7c)
3-[2-[tert-butyl(dimethyl)silyl]oxyethyl]-2-fluoro-benzaldehyde
将2,2,6,6-四甲基吡啶(39.1,277mmol)加入THF(200mL)中,-78℃缓慢滴加正丁基锂(0.133L,160g/L),搅拌15min后,滴加刚7b(35.2,138mmol)的THF溶液(100mL),-78℃搅拌2h。加入DMF(31mL)和THF(50mL)组成的混合溶液,保持-78℃搅拌反应1h,后室温反应18h。将反应液倒入0.5M盐酸水溶液(1000mL)中,并滴加3.0M盐酸水溶液至pH≈7,用乙酸乙酯(300mL×3)萃取,合并后的有机相用无水硫酸钠干燥,减压浓缩后得到深红色液体状的7c(38.6g,产率:99%)。
1H NMR(400MHz,CDCl3)δ10.36(s,1H),7.74–7.69(m,1H),7.53-7.48(m,1H),7.23–7.13(m,1H),7.06–6.95(m,1H),3.84(t,2H),2.94-2.89(m,2H),0.84(d,9H),-0.04(d,6H).
第三步:叔丁基-[2-[3-[[4-(4-乙基-2-吡啶基)-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-基]甲基]-2-氟苯基]乙氧基]-二甲基硅烷(7d)
tert-butyl-[2-[3-[[4-(4-ethyl-2-pyridyl)-1-oxa-4,9-diazaspiro[5.5]undecan-9-yl]methyl]-2-fluoro-phenyl]etho xy]-dimethyl-silane
将4-(4-乙基-2-吡啶基)-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-羧酸叔丁酯(16C)(1.80g,4.98mmol)加入DCM(20mL),0℃滴加三氟乙酸(5mL),室温反应1小时,将反应液减压浓缩,浓缩后的残余物加入DCM(15mL)和甲醇(10mL)的混合溶剂中,加入7c(1.69g,5.97mmol),室温搅拌30分钟后加入三乙酰氧基硼氢化钠(3.16g,14.9mmol),室温搅拌3小时。加入饱和碳酸氢钠水溶液(50mL),用DCM(100mL×2)萃取,合并后的有机相用无水硫酸钠干燥,减压浓缩后柱层析分离(DCM:甲醇(v/v)=1:0~15:1)得黄色液体状的7d(1.36g,产率52%)。
第四步:2-[3-[[4-(4-甲基-2-吡啶基)-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-基]甲基]-2-氟-苯基]乙醇(7e)
2-[3-[[4-(4-ethyl-2-pyridyl)-1-oxa-4,9-diazaspiro[5.5]undecan-9-yl]methyl]-2-fluoro-phenyl]ethanol
将7d(1.3g,2.5mmol)溶于THF(15mL)中,加入四丁基氟化铵(1.3g,4.9mmol),室温搅拌1小时。加入水(50mL),用DCM(50mL×2)萃取,合并后的有机相用无水硫酸钠干燥,减压浓缩后得到黄色液体状的7e(0.89g,产率87%)。
第四步:3-[2-[3-[[4-(4-甲基-2-吡啶基)-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-基]甲基]-2-氟苯基]乙氧基]
丙烯酸叔丁酯(中间体7)
tert-butyl 3-[2-[3-[[4-(4-ethyl-2-pyridyl)-1-oxa-4,9-diazaspiro[5.5]undecan-9-yl]methyl]-2-fluoro-phenyl]ethoxy]propanoate
将7e(0.89g,2.2mmol)加入乙腈(20mL)中,加入丙烯酸叔丁酯(0.83g,6.5mmol)和苄基三甲基氢氧化铵(0.27g,0.65mmol),40℃搅拌3小时。加入水(50mL),用DCM(100mL×2)萃取,合并后的有机相用无水硫酸钠干燥,减压浓缩后柱层析分离(DCM/甲醇(v/v)=15:1)得到黄色液体状的中间体7(0.94g,产率81%)。
中间体8:4-吡嗪基-2-基-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-羧酸叔丁酯
tert-butyl 4-pyrazin-2-yl-1-oxa-4,9-diazaspiro[5.5]undecane-9-carboxylate
将1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-羧酸叔丁酯(0.824g,4.0mmol)和2-碘吡嗪(8a)(1.03g,4.0mmol)加入DMF(20mL),110℃搅拌8小时。反应液冷却至室温,加入水(100mL),用乙酸乙酯(200mL×2)萃取,合并后的有机相用饱和食盐水洗(100mL),无水硫酸钠干燥,减压浓缩后柱层析分离(石油醚/乙酸乙酯(v/v)=1:1)得到黄色液体状的中间体8(0.4g,产率30%)。
LCMS m/z=335.2.[M+1]。
中间体9:4-嘧啶-2-基-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-羧酸叔丁酯
tert-butyl 4-pyrimidin-2-yl-1-oxa-4,9-diazaspiro[5.5]undecane-9-carboxylate
将1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-羧酸叔丁酯(1.03,4.0mmol)溶于DMF(20mL)中,依次加入2-氯嘧啶(9a)(0.46g,1.0mmol)、碳酸钾(1.11g,8.0mmol),50℃搅拌4小时。加入水(100mL),用乙酸乙酯(200mL,100mL)萃取,合并后的有机相用饱和食盐水洗(100mL),无水硫酸钠干燥,减压浓缩后柱层析分离(石油醚/乙酸乙酯(v/v)=1:1)得到黄色液体状的中间体9(1.1.34g,产率100%)。
LCMS m/z=335.2[M+1]。
中间体10:4-噻唑-2-基-1-氧杂-4,9-二氮杂螺[5.5]-十一烷-9-羧酸叔丁酯
tert-butyl 4-thiazol-2-yl-1-oxa-4,9-diazaspiro[5.5]undecane-9-carboxylate
将1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-羧酸叔丁酯(1.03g,4.0mmol)溶于DMF(20mL)中,依次加入2-溴-噻唑(10a)(0.655g,4.0mmol)、碳酸铯(2.61g,8.0mmol),50℃搅拌4小时。加入水(100mL),用乙
酸乙酯(200mL,100mL)萃取,合并后的有机相用饱和食盐水(100mL)洗涤,无水硫酸钠干燥,减压浓缩后柱层析分离(石油醚/乙酸乙酯(v/v)=1:1)得到黄色液体状的中间体10(0.28g,产率21%)。
LCMS m/z=340.1[M+1]。
中间体11:4-(3-吡啶基)-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-羧酸叔丁酯
tert-butyl 4-(3-pyridyl)-1-oxa-4,9-diazaspiro[5.5]undecane-9-carboxylate
将1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-羧酸叔丁酯(2.56g,10.0mmol)溶于1,4-二氧六环(50mL)中,加入3-氯吡啶(11a)(1.70g,15.0mmol),叔丁醇钠(2.4g,25.0mmol),4,5-双二苯基膦-9,9-二甲基氧杂蒽(0.8g,1.4mmol),醋酸钯(0.16g,0.7mmol),氮气氛回流8小时。反应液冷却至室温,加入水(100mL),用乙酸乙酯(100mL×2)萃取,合并后的有机相用饱和食盐水洗(100mL),无水硫酸钠干燥,减压浓缩后柱层析分离(石油醚/乙酸乙酯(v/v)=1:1)得到黄色液体状的中间体11(1.1g,产率33%)。
中间体12:4-(4-硝基-2-吡啶基)-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-羧酸叔丁酯
tert-butyl 4-(4-nitro-2-pyridyl)-1-oxa-4,9-diazaspiro[5.5]undecane-9-carboxylate
将1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-羧酸叔丁酯(1.28g,4.99mmol)溶于乙腈(5mL)中,加入2-氯-4-硝基吡啶(12a)(0.87g,5.49mmol),三乙胺(1.52g,15.0mmol),置入微波反应器后反应140℃反应2小时。反应液冷却至室温,直接减压浓缩后柱层析分离(石油醚/乙酸乙酯(v/v)=1:1)得到黄色液体状的中间体12(0.81g,产率43%)。
中间体13:4-(4-氟-2-吡啶基)-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-羧酸叔丁酯
tert-butyl 4-(4-fluoro-2-pyridyl)-1-oxa-4,9-diazaspiro[5.5]undecane-9-carboxylate
将1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-羧酸叔丁酯(1.28g,4.99mmol)溶于1,4-二氧六环(20mL)中,加入2-氯-4-氟吡啶(13a)(0.72g,5.5mmol),叔丁醇钠(1.2g,12.5mmol),4,5-双二苯基膦-9,9-二甲基氧杂蒽(0.41g,0.7mmol),醋酸钯(0.08g,0.35mmol),氮气氛回流8小时后。反应液冷却至室温,加入水(100mL),用乙酸乙酯(100mL×2)萃取,合并后的有机相用饱和食盐水洗(100mL),无水硫酸钠干燥,减压浓缩后柱层析分离(石油醚/乙酸乙酯(v/v)=1:1)得到黄色液体状的中间体13(0.68g,产率39%)。
中间体14:4-(3-硝基-2-吡啶基)-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-羧酸叔丁酯
tert-butyl 4-(3-nitro-2-pyridyl)-1-oxa-4,9-diazaspiro[5.5]undecane-9-carboxylate
将1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-羧酸叔丁酯(1.28g,4.99mmol)溶于DMF(20mL)中,加入2-氯-3-硝基吡啶(14a)(1.19g,7.5mmol),碳酸钾(1.38g,10.0mmol),80℃反应8小时。反应液冷却至室温,直接减压浓缩后柱层析分离(石油醚/乙酸乙酯(v/v)=1:1)得到黄色液体状的中间体14(0.95g,产率50%)。
中间体15:4-(4-环丙基-2-吡啶基)-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-羧酸叔丁酯
tert-butyl 4-(4-cyclopropyl-2-pyridyl)-1-oxa-4,9-diazaspiro[5.5]undecane-9-carboxylate
将1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-羧酸叔丁酯(1.20g,4.68mmol)溶于1,4-二氧六环(80mL)中,加入2-氯-4-环丙基吡啶(15a)(0.79g,5.15mmol),叔丁醇钠(1.12g,11.7mmol),4,5-双二苯基膦-9,9-二甲基氧杂蒽(0.38g,0.65mmol),醋酸钯(0.07g,0.33mmol),氮气氛回流8小时。反应液冷却至室温,加入水(100mL),用乙酸乙酯(100mL×2)萃取,合并后的有机相用饱和食盐水洗(100mL),无水硫酸钠干燥,减压浓缩后柱层析分离(石油醚/乙酸乙酯(v/v)=1:1)得到黄色液体状的中间体15(0.95g,产率54%)。
LCMS m/z=374.2[M+1].
中间体16:4-[4-(三氟甲基)-2-吡啶基]-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-羧酸叔丁酯
tert-butyl 4-[4-(trifluoromethyl)-2-pyridyl]-1-oxa-4,9-diazaspiro[5.5]undecane-9-carboxylate
将1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-羧酸叔丁酯(5.3g,21mmol)溶于1,4-二氧六环(50mL)中,加入2-氯-4-(三氟甲基)吡啶(16a)(4.1g,23mmol),叔丁醇钠(5.0g,52.0mmol),4,5-双二苯基膦-9,9-二甲基氧杂蒽(1.7g,2.9mmol),醋酸钯(0.3g,1.4mmol),氮气氛回流8小时。反应液冷却至室温,加入水(100mL),用乙酸乙酯(100mL×2)萃取,合并后的有机相用饱和食盐水洗(100mL),无水硫酸钠干燥,减压浓缩后柱层析分离(石油醚/乙酸乙酯(v/v)=1:1)得到黄色液体状的中间体16(5.4g,产率65%)。
LCMS m/z=402.2[M+1].
中间体17:4-[4-(2-噻吩)嘧啶-2-基]-1-氧杂-4,9-二氮杂螺环[5.5]十一烷-9-羧酸叔丁酯
tert-butyl 4-[4-(2-thienyl)pyrimidin-2-yl]-1-oxa-4,9-diazaspiro[5.5]undecane-9-carboxylate
第一步:2-氯-4-(2-噻吩)嘧啶(17b)
2-chloro-4-(2-thienyl)pyrimidine
将2,4-二氯嘧啶(3.0g,20.1mmol),2-噻吩-硼酸(17a)(3.1g,24.2mmol)溶于1,4-二氧六环(50ml)以及水(10ml)的混合溶剂中,加入碳酸钠(4.26g,40.2mmol),[1,1'-双(二苯基膦基)二茂铁]二氯化钯(0.73g,1.0mmol),氮气氛90℃反应5小时。反应液冷却至室温,加入饱和碳酸氢钠水溶液(50mL)淬灭反应,用乙酸乙酯(100mL×2)萃取,合并后的有机相用饱和食盐水(50mL)洗涤,无水硫酸钠干燥,减压浓缩后柱层析分离(石油醚/乙酸乙酯(v/v)=5:1)得到棕色液体状的17b(2.6g,产率63.4%)。
LCMS m/z=197.6[M+1]。
第二步:4-[4-(2-噻吩)嘧啶-2-基]-1-氧杂-4,9-二氮杂螺环[5.5]十一烷-9-羧酸叔丁酯
tert-butyl-4-[4-(2-thienyl)pyrimidin-2-yl]-1-oxa-4,9-diazaspiro[5.5]undecane-9-carboxylate
将17b(2.6g,12.75mmol),1-氧杂-4,9-二氮杂螺环[5.5]十一烷-9-羧酸叔丁酯(3.42g,13.38mmol)加入DMF(50mL)中,加入碳酸钾(5.3g,38.25mmol),75℃反应5小时。反应液冷却至室温,加入饱和碳酸氢钠水溶液(50mL),用乙酸乙酯(100mL×2)萃取,合并后的有机相用饱和食盐水(50mL)洗,无水硫酸钠干燥,减压浓缩后柱层析分离(石油醚/乙酸乙酯(v/v)=5:1)得到棕色液体状的中间体17(5.0g,产率92.6%)。
中间体18:4-[4-苯氧嘧啶-2-基]-1-氧杂-4,9-二氮杂螺环[5.5]十一烷-9-羧酸叔丁酯
tert-butyl-4-(4-phenoxypyrimidin-2-yl)-1-oxa-4,9-diazaspiro[5.5]undecane-9-carboxylate
第一步:2-氯-4-苯氧基-嘧啶(18b)
2-chloro-4-phenoxy-pyrimidine
将2,4-二氯嘧啶(18a)(5.0g,33.56mmol)和苯酚(3.15g,33.56mmol)溶于DMF(50ml)中,加入碳酸钾(6.9g,50.3mmol),90℃反应5小时。反应液冷却是室温,加入水溶液(100mL),用乙酸乙酯(100mL×2)萃取,合并后的有机相用饱和食盐水(50mL)洗涤,无水硫酸钠干燥,减压浓缩后柱层析分离(石油醚/乙酸乙酯(v/v)=5:1)得到棕色液体状的18b(5.0g,产率72.5%)。
LCMS m/z=207.6[M+1]。
第二步:4-[4-苯氧嘧啶-2-基]-1-氧杂-4,9-二氮杂螺环[5.5]十一烷-9-羧酸叔丁酯(中间体18)
tert-butyl-4-(4-phenoxypyrimidin-2-yl)-1-oxa-4,9-diazaspiro[5.5]undecane-9-carboxylate
将2b(1.0g,4.8mmol)和1-氧杂-4,9-二氮杂螺环[5.5]十一烷-9-羧酸叔丁酯(1.29g,5.04mmol)加入DMF(25mL)中,加入碳酸钾(2g,14.4mmol),75℃反应5小时。反应液冷却至室温,加入饱和碳酸氢钠水溶液(50mL),用乙酸乙酯(100mL×2)萃取,合并后的有机相用饱和食盐水(50mL)洗涤,无水硫酸钠干燥,减压浓缩后柱层析分离(石油醚/乙酸乙酯(v/v)=5:1)得到棕色液体状的中间体18(1.5g,产率72.8%)。
LCMS m/z=427.5[M+1]。
中间体19:4-[4-苯胺嘧啶-2-基]-1-氧杂-4,9-二氮杂螺环[5.5]十一烷-9-羧酸叔丁酯
tert-butyl 4-(4-anilinopyrimidin-2-yl)-1-oxa-4,9-diazaspiro[5.5]undecane-9-carboxylate
第一步:2-氯-4-苯胺基-嘧啶19b
2-chloro-N-phenyl-pyrimidin-4-amine
将2,4-二氯嘧啶18a(5.0g,33.56mmol)和苯胺(3.15g,33.56mmol)溶于乙醇(50ml)中,加入三乙胺(5.1g,50.3mmol),90℃反应5小时。反应液减压浓缩,加入水溶液(100mL),用乙酸乙酯(100mL×2)萃取,合并后的有机相用饱和食盐水(50mL)洗涤,无水硫酸钠干燥,减压浓缩后柱层析分离(石油醚/乙酸乙酯(v/v)=5:1)得到棕色液体状的19b(4.5g,产率65.3%)。
LCMS m/z=206.6[M+1]。
第二步:4-[4-苯胺嘧啶-2-基]-1-氧杂-4,9-二氮杂螺环[5.5]十一烷-9-羧酸叔丁酯(中间体19)
tert-butyl 4-(4-anilinopyrimidin-2-yl)-1-oxa-4,9-diazaspiro[5.5]undecane-9-carboxylate
将2-氯-4-苯胺嘧啶(3b)(1.0g,4.8mmol)和1-氧杂-4,9-二氮杂螺环[5.5]十一烷-9-羧酸叔丁酯(1.29g,5.04mmol)加入DMF(25mL)中,加入碳酸钾(2g,14.4mmol),75℃反应5小时。反应液冷却至室温,加入饱和碳酸氢钠水溶液(50mL),用乙酸乙酯(100mL×2)萃取,合并后的有机相用饱和食盐水(50mL)洗涤,无水硫酸钠干燥,减压浓缩后柱层析分离(石油醚/乙酸乙酯(v/v)=5:1)得到棕色液体状的中间体19(1.4g,产率67.9%)。
LCMS m/z=426.5[M+1]。
中间体20:4-[5-甲基嘧啶-2-基]-1-氧杂-4,9-二氮杂螺环[5.5]十一烷-9-羧酸叔丁酯
tert-butyl 4-(5-methylpyrimidin-2-yl)-1-oxa-4,9-diazaspiro[5.5]undecane-9-carboxylate
将2-氯-5-甲基嘧啶(20b)(1.0g,7.8mmol)和1-氧杂-4,9-二氮杂螺环[5.5]十一烷-9-羧酸叔丁酯(2.1g,8.2mmol)加入DMF(25mL)中,加入碳酸钾(3.2g,23.4mmol),90℃反应5小时。加入水溶液(100mL),用乙酸乙酯(50mL×2)萃取,合并后的有机相用饱和食盐水(50mL)洗涤,无水硫酸钠干燥,减压浓缩后柱层析分离(石油醚/乙酸乙酯(v/v)=5:1)得到棕色液体状的中间体20(2.0g,产率74.0%)。
LCMS m/z=349.4[M+1]。
中间体21:4-[4-异丙氧基嘧啶-2-基]-1-氧杂-4,9-二氮杂螺环[5.5]十一烷-9-羧酸叔丁酯
tert-butyl 4-(4-isopropoxypyrimidin-2-yl)-1-oxa-4,9-diazaspiro[5.5]undecane-9-carboxylate
将2-氯-4-异丙氧基嘧啶(5a)(0.5g,2.9mmol)和1-氧杂-4,9-二氮杂螺环[5.5]十一烷-9-羧酸叔丁酯(0.77g,3mmol)加入DMF(25mL)中,加入碳酸钾(1.2g,8.7mmol),90℃反应5小时。加入水溶液(100mL),用乙酸乙酯(50mL×2)萃取,合并后的有机相用饱和食盐水(50mL)洗涤,无水硫酸钠干燥,减压浓缩后柱层析分离(石油醚/乙酸乙酯(v/v)=5:1)得到棕色液体状的中间体21(0.9g,产率78.9%)。
LCMS m/z=393.2[M+1]。
中间体22:4-(4-异丙基-2-吡啶基)-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-羧酸叔丁酯
tert-butyl 4-(4-isopropyl-2-pyridyl)-1-oxa-4,9-diazaspiro[5.5]undecane-9-carboxylate
将1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-羧酸叔丁酯(2.56g,10.0mmol)加入1,4-二氧六环(50mL)中,加入2-氯-4-异丙基吡啶22a(1.87g,12.0mmol),叔丁醇钠(2.4g,25.0mmol),4,5-双二苯基膦-9,9-二甲基氧杂蒽(0.81g,1.4mmol)和醋酸钯(0.16g,0.7mmol),氮气氛回流8小时后。加入水(100mL),用乙酸乙酯(100mL×2)萃取,合并后的有机相用饱和食盐水(100mL)洗涤,无水硫酸钠干燥,减压浓缩后柱层析分离(石油醚/乙酸乙酯(v/v)=1:1)得到黄色液体状的中间体22(2.5g,产率67%)。
中间体23:4-[4-(1-羟基-1-甲基乙基)嘧啶-2-基]-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-羧酸叔丁酯
tert-butyl 4-[4-(1-hydroxy-1-methyl-ethyl)pyrimidin-2-yl]-1-oxa-4,9-diazaspiro[5.5]undecane-9-carboxylate
第一步:2-(2-氯嘧啶-4-基)丙烷-2-醇(23b)
2-(2-chloropyrimidin-4-yl)propan-2-ol
将2-氯嘧啶-4-甲酸甲酯(23a)(1.00g,5.79mmol)加入THF(30mL),氮气保护下,-78℃缓慢加入甲基溴化镁的THF溶液(6mL,3mol/L),室温反应1.5小时。加入水(100mL),用DCM(100mL×2)萃取,合并后的有机相用饱和食盐水洗(100mL),无水硫酸钠干燥,减压浓缩后柱层析分离(石油醚/乙酸乙酯(v/v)=10:1)得到褐色液体状的23b(0.63g,产率63%)。
第二步:4-[4-(1-羟基-1-甲基乙基)嘧啶-2-基]-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-羧酸叔丁酯(中间体23)
tert-butyl 4-[4-(1-hydroxy-1-methyl-ethyl)pyrimidin-2-yl]-1-oxa-4,9-diazaspiro[5.5]undecane-9-carboxylate
将23b(0.95g,3.7mmol)加入DMF(50mL)中,加入23b(0.61g,3.5mmol),碳酸钾(0.98g,7.1mmol),氮气氛75℃反应5小时。加入水(100mL),用DCM(100mL×2)萃取,合并后的有机相用饱和食盐水洗(100mL),无水硫酸钠干燥,减压浓缩后得到黄色液体状的中间体23(1.01g,产率73%)。
中间体24:4-(4-苯基嘧啶-2-基)-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-羧酸叔丁酯
tert-butyl 4-(4-phenylpyrimidin-2-yl)-1-oxa-4,9-diazaspiro[5.5]undecane-9-carboxylate
第一步:2-氯-4-苯基嘧啶(24b)
2-chloro-4-phenyl-pyrimidine
将24a(1.5g,10.0mmol)加入1,4-二氧六环(52mL)和水(13mL)的混合溶剂,加入苯硼酸(1.8g,15.0mmol),碳酸钠(2.7g,25.0mmol),PdCl2(dppf)(cas:72287-26-4)(0.37g,0.5mmol)。90℃反应5小时。将反应液用硅藻土过滤,向滤液中加入水(100mL),用乙酸乙酯(50mL×3)萃取,合并后的有机相用饱和食盐水(100mL)洗涤,无水硫酸钠干燥,减压浓缩后柱层析分离(石油醚/乙酸乙酯(v/v)=10:1)得到白色固体状的24b(1.32g,产率70%)。
第二步:4-(4-苯基嘧啶-2-基)-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-羧酸叔丁酯(中间体24)
tert-butyl 4-(4-phenylpyrimidin-2-yl)-1-oxa-4,9-diazaspiro[5.5]undecane-9-carboxylate
将1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-羧酸叔丁酯(1.86g,7.3mmol)加入DMF(50mL)中,加入
24b(1.32g,6.9mmol),碳酸钾(1.91g,13.8mmol),氮气氛75℃反应5小时。加入水(100mL),用DCM(100mL×2)萃取,合并后的有机相用饱和食盐水(100mL)洗涤,无水硫酸钠干燥,减压浓缩后得到黄色液体状的中间体24(2.01g,产率71%)。
中间体25:4-(4-甲氧基苯基)-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-羧酸叔丁酯
tert-butyl 4-(4-methoxyphenyl)-1-oxa-4,9-diazaspiro[5.5]undecane-9-carboxylate
将1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-羧酸叔丁酯(1.28g,5.0mmol)加入1,4-二氧六环(50mL)中,加入1-溴-4-甲氧基苯25a(1.4g,7.5mmol),叔丁醇钠(1.2g,12.5mmol),4,5-双二苯基膦-9,9-二甲基氧杂蒽(0.4g,0.7mmol),醋酸钯(0.08g,0.35mmol),氮气氛回流8小时。加入水(100mL),用乙酸乙酯(100mL×2)萃取,合并后的有机相用饱和食盐水(100mL)洗涤,无水硫酸钠干燥,减压浓缩后柱层析分离(石油醚/乙酸乙酯(v/v)=1:1)得到黄色液体状的中间体25(0.96g,产率53%)。
中间体26:4-(1-甲基吡唑-4-基)-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-羧酸叔丁酯
tert-butyl 4-(1-methylpyrazol-4-yl)-1-oxa-4,9-diazaspiro[5.5]undecane-9-carboxylate
将1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-羧酸叔丁酯(1.28g,5.0mmol)加入1,4-二氧六环(50mL)中,加入4-溴-1-甲基吡唑(1.21g,7.49mmol),叔丁醇钠(1.2g,12.5mmol),三叔丁基膦(0.1g,0.5mmol),醋酸钯(0.08g,0.35mmol),氮气氛回流8小时。加入水(100mL),用乙酸乙酯(100mL×2)萃取,合并后的有机相用饱和食盐水(100mL)洗涤,无水硫酸钠干燥,减压浓缩后柱层析分离(石油醚/乙酸乙酯(v/v)=4:1)得到黄色液体状的中间体26(1.0g,产率50%)。
中间体27:4-(3-氰基-2-吡啶基)-1-氧-4,9-二氮杂螺[5.5]十一烷-9-羧酸叔丁酯
tert-butyl 4-(3-cyano-2-pyridyl)-1-oxa-4,9-diazaspiro[5.5]undecane-9-carboxylate
将1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-甲酸叔丁酯(1.28g,5.0mmol)加入THF(50mL)中,加入2-氟吡啶-3-腈(0.92g,7.5mmol),三乙胺(1.52g,15.0mmol),80℃回流8小时。反应液冷却至室温,减压浓缩后柱层析分离(石油醚/乙酸乙酯(v/v)=1:1)得到黄色液体状的中间体27(1.7g,产率95%)。
中间体28:4-(2-氯苯基)-1-氧-4,9-二氮杂螺[5.5]十一烷-9-羧酸叔丁酯
tert-butyl 4-(2-chlorophenyl)-1-oxa-4,9-diazaspiro[5.5]undecane-9-carboxylate
将1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-羧酸叔丁酯(1.28g,5.0mmol)加入1,4-二氧六环(50mL)中,加入1-溴-2-氯苯(1.43g,7.5mmol),叔丁醇钠(1.2g,12.5mmol),4,5-双二苯基膦-9,9-二甲基氧杂蒽(0.40g,0.7mmol),醋酸钯(0.08g,0.35mmol),氮气氛100℃反应8小时。加入水(100mL),用乙酸乙酯(100mL×2)萃取,合并后的有机相用饱和食盐水(100mL)洗涤,无水硫酸钠干燥,减压浓缩后柱层析分离(石油醚/乙酸乙酯(v/v)=1:1)得到黄色液体状的中间体28(1.2g,产率65%)。
中间体29:[2-(1-氧杂-4,9-二氮杂[5.5]十一烷-4-基)-4-吡啶基]甲醇
[2-(1-oxa-4,9-diazaspiro[5.5]undecan-4-yl)-4-pyridyl]methanol
第一步:4-[4-[[叔丁基(二甲基)硅基]氧甲基]-2-吡啶基]-1-氧杂-4,9-二氮杂螺环[5.5]十一烷-9-羧酸叔丁酯(29b)
tert-butyl 4-[4-[[tert-butyl(dimethyl)silyl]oxymethyl]-2-pyridyl]-1-oxa-4,9-diazaspiro[5.5]undecane-9-carboxylate
将1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-羧酸叔丁酯(1.64g,6.4mmol)加入1,4-二氧六环(30mL)中,加入4-(((叔丁基甲基硅基)氧)甲基)-2-氯吡啶(29a)(参考WO2008074752A2制备得到)(1.65g,6.4mmol),叔丁醇钠(1.54g,16.0mmol),4,5-双二苯基膦-9,9-二甲基氧杂蒽(0.26g,0.45mmol)和醋酸钯(0.072g,0.32mmol),氮气氛回流8小时。待反应冷至室温后,加入水(100mL),用乙酸乙酯(200mL×2)萃取,合并后的有机相用饱和食盐水(100mL)洗涤,无水硫酸钠干燥,减压浓缩后柱层析分离(石油醚/乙酸乙酯(v/v)=1:1)得到黄色固体状的29b(2.0g,产率65%)。
LCMS m/z=478.2[M+1].
第二步:[2-(1-氧杂-4,9-二氮杂[5.5]十一烷-4-基)-4-吡啶基]甲醇(中间体29)
[2-(1-oxa-4,9-diazaspiro[5.5]undecan-4-yl)-4-pyridyl]methanol
将29b(1.7g,3.56mmol)溶于DCM(10mL)中,加入三氟乙酸(3mL),室温搅拌2小时。减压浓缩除去反应溶剂,向残余物中加入甲苯(5mL),减压浓缩后得到黄色液体状的中间体29的三氟乙酸盐(1.34,产率100%)。
LCMS m/z=264.3[M+1].
中间体30:4-(2-氯-4-吡啶基)-1-氧杂-4,9-二氮杂螺环[5.5]十一烷-9-羧酸叔丁酯
tert-butyl 4-(2-chloro-4-pyridyl)-1-oxa-4,9-diazaspiro[5.5]undecane-9-carboxylate
将1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-甲酸叔丁酯(2.56g,10.0mmol)加入1,4-二氧六环(50mL)中,依次加入2-氯-4-溴-吡啶(30a)(2.1g,11.0mmol),叔丁醇钠(2.4g,25.0mmol),4,5-双二苯基膦-9,9-二甲基氧杂蒽(0.58g,1.0mmol)和醋酸钯(0.11g,0.5mmol),通过氮气置换后,氮气氛回流8小时。待反应冷至室温后,加入水(100mL),用乙酸乙酯(200mL×2)萃取,合并后的有机相用饱和食盐水(100mL)洗涤,无水硫酸钠干燥,减压浓缩后柱层析分离(石油醚/乙酸乙酯(v/v)=1:1)得到黄色固体状的中间体30(2.9g,产率80%)。
LCMS m/z=368.2[M+1]。
中间体31:4-(4-甲氧基-2-吡啶基)-1-氧杂-4,9-二氮杂螺环[5.5]十一烷-9-羧酸叔丁酯
tert-butyl 4-(4-methoxy-2-pyridyl)-1-oxa-4,9-diazaspiro[5.5]undecane-9-carboxylate
将1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-羧酸叔丁酯(5.12g,20.0mmol)加入1,4-二氧六环(60mL)中,加入2-氯-4-甲氧基-吡啶(4.3g,30.0mmol),碳酸铯(13.0g,40.0mmol),4,5-双二苯基膦-9,9-二甲基氧杂蒽(1.6g,2.8mmol)和醋酸钯(0.3g,1.4mmol),氮气氛回流8小时。待反应冷至室温后,加入水(100mL),用乙酸乙酯(200mL×2)萃取,合并后的有机相用饱和食盐水(100mL)洗涤,无水硫酸钠干燥,减压浓缩,残留物经硅胶柱层析纯化(乙酸乙酯/石油醚(v/v)=1:2),得到黄色油状的中间体31(6.5g,产率90%)。
中间体32:4-苯基-1-氧杂-4,9-二氮杂螺环[5.5]十一烷-9-羧酸叔丁酯
tert-butyl 4-phenyl-1-oxa-4,9-diazaspiro[5.5]undecane-9-carboxylate
将1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-羧酸叔丁酯(5.12g,20.0mmol)加入1,4-二氧六环(50mL)中,加入溴苯(4.7g,30.0mmol),叔丁醇钠(3.84g,40.0mmol),4,5-双二苯基膦-9,9-二甲基氧杂蒽(1.6g,2.8mmol)和醋酸钯(0.3g,1.4mmol),氮气氛回流8小时。待反应冷至室温后,加入水(100mL),用乙酸乙酯(200mL×2)萃取,合并后的有机相用饱和食盐水(100mL)洗涤,无水硫酸钠干燥,减压浓缩,减压浓缩,残留物经硅胶柱层析纯化(乙酸乙酯/石油醚(v/v)=1:10),得到黄色油状的中间体32(4.6g,产率69%)。
中间体33:4-(4-吡啶基)-1-氧杂-4,9-二氮杂螺环[5.5]十一烷-9-羧酸叔丁酯
tert-butyl 4-(4-pyridyl)-1-oxa-4,9-diazaspiro[5.5]undecane-9-carboxylate
将1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-羧酸叔丁酯(5.12g,20.0mmol)加入1,4-二氧六环(50mL)中,加入4-氯-吡啶(3.4g,30.0mmol),叔丁醇钠(3.84g,40.0mmol),4,5-双二苯基膦-9,9-二甲基氧杂蒽(1.6g,2.8mmol)和醋酸钯(0.3g,1.4mmol),氮气氛回流10小时。待反应冷至室温后,加入水(100mL),用乙酸乙酯(200mL×2)萃取,合并后的有机相用饱和食盐水(100mL)洗涤,无水硫酸钠干燥,减压浓缩,残留物经硅胶柱层析纯化(乙酸乙酯/石油醚(v/v)=1:1),得到黄色油状的中间体33(3.8g,产率57%)。
中间体34:4-(4-氟-2-吡啶基)-1-氧杂-4,9-二氮杂螺环[5.5]十一烷-9-羧酸叔丁酯
tert-butyl 4-(4-fluoro-2-pyridyl)-1-oxa-4,9-diazaspiro[5.5]undecane-9-carboxylate
将1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-羧酸叔丁酯(1.28g,5.0mmol)加入1,4-二氧六环(30mL)中,加入2-氯-4-氟-吡啶(0.72g,5.5mmol),叔丁醇钠(1.2g,12.5mmol),4,5-双二苯基膦-9,9-二甲基氧杂蒽(0.41g,0.7mmol),醋酸钯(0.079g,0.35mmol),氮气氛回流8小时。待反应冷至室温后,加入水(100mL),用乙酸乙酯(200mL×2)萃取,合并后的有机相用饱和食盐水(100mL)洗涤,无水硫酸钠干燥,减压浓缩后柱层析分离(石油醚/乙酸乙酯(v/v)=1:1)得到黄色固体状的中间体34(0.68g,产率39%)。
LCMS m/z=352.3[M+1].
中间体35:4-(4-异丙基苯基)-1-氧杂-4,9-二氮杂螺环[5.5]十一烷-9-羧酸叔丁酯
tert-butyl 4-(4-isopropylphenyl)-1-oxa-4,9-diazaspiro[5.5]undecane-9-carboxylate
将1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-羧酸叔丁酯(1.28g,5.0mmol)加入1,4-二氧六环(30mL)中,加入1-氯-4-异丙基苯(35a)(1.1g,5.5mmol),叔丁醇钠(1.2g,12.5mmol),4,5-双二苯基膦-9,9-二甲基氧杂蒽(0.41g,0.7mmol),醋酸钯(0.079g,0.35mmol),氮气氛回流8小时。待反应冷至室温后,加入水(100mL),用乙酸乙酯(200mL×2)萃取,合并后的有机相用饱和食盐水(100mL)洗涤,无水硫酸钠干燥,减压浓缩后柱层析分离(石油醚/乙酸乙酯(v/v)=1:1)得到黄色固体状的中间体35(1.68g,产率90%)。
LCMS m/z=375.3[M+1].
中间体36:4-(3-苯甲基氧基羰基-2-吡啶基)-1-氧杂-4,9-二氮杂螺环[5.5]十一烷-9-羧酸叔丁酯
tert-butyl 4-(3-benzyloxycarbonyl-2-pyridyl)-1-oxa-4,9-diazaspiro[5.5]undecane-9-carboxylate
将1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-羧酸叔丁酯(1.3g,5.0mmol)置于微波反应管(20mL)中,加入1,4-二氧六环(10mL),36a(1.2g,5.0mmol),N,N-二异丙基乙基胺(1.9g,15.0mmol),130℃微波反应2小时。待反应冷至室温,减压浓缩反应液后柱层析分离(石油醚/乙酸乙酯(v/v)=1:1)得到黄色固体状的中间体36(1.8g,产率77%)。
LCMS m/z=468.3[M+1]。
中间体8~中间体28,中间体30~中间体36脱Boc反应条件:底物溶于溶于DCM中,加入三氟乙酸(为DCM体积的1/5~1/2),室温反应至反应结束。反应液直接减压浓缩,得到对应的脱Boc产物的三氟乙酸盐。
实施例1:3-[2-[3-[2-[4-[1-(环丙甲基)吡唑-3-羰基]-1-氧杂-4,9-杂螺[5.5]十一烷-9-基]乙基]苯基]乙氧基]-N-[2-[[(2R)-2-羟基-2-(8-羟基-2-氧代-1H-喹啉-5-基)乙基]胺基]乙基]-N-甲基-丙酰胺(化合物1)
3-[2-[3-[2-[4-[1-(cyclopropylmethyl)pyrazole-3-carbonyl]-1-oxa-4,9-diazaspiro[5.5]undecan-9-yl]ethyl]phenyl]ethoxy]-N-[2-[[(2R)-2-hydroxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]-N-methyl-propanamide
第一步:1-(环丙基甲基)吡唑-3-羧酸乙酯(1B)
ethyl 1-(cyclopropylmethyl)pyrazole-3-carboxylate
称取钠氢(5.7g,143.7mmol,60%w/w)加入DMF(100mL)中,0℃滴加1A(10.0g,71.4mmol)的DMF溶液(100mL),滴加完毕后,0℃搅拌30分钟,滴加溴甲基环丙烷(14.5g,107.0mmol),室温搅拌3小时。滴加水淬灭反应(150mL),用乙酸乙酯(150mL×2)萃取,合并后的有机相用饱和食盐水(100mL×1)洗涤,无水硫酸钠干燥,过滤,滤液减压浓缩后柱层析分离(石油醚/乙酸乙酯(v/v)=2:1)得到黄色液体状的1B(6.5g,产率47%)。
1H NMR(400MHz,CDCl3)δ7.52-7.51(d,1H),6.80-6.79(d,1H),4.40-4.35(m,2H),4.05-4.04(d,2H),1.37-1.32(m,3H),1.26-1.23(m,1H),0.64-0.62(m,2H),0.37-0.35(m,2H).
第二步:1-(环丙基甲基)吡唑-3-羧酸(1C)
1-(cyclopropylmethyl)pyrazole-3-carboxylic acid
向1B(3.86g,20.0mmol)中加入THF(60mL)、甲醇(15mL)和水(15mL),搅拌均匀,加入氢氧化锂(0.95g,40.0mmol),室温搅拌3小时。反应液直接减压浓缩除去大部分THF和甲醇,加入水(100mL),稀盐酸(1.0M)调节水相pH至6,乙酸乙酯萃取(150mL×2),合并后的有机相用无水硫酸钠干燥,过滤,滤液减压浓缩后得到白色固体状的1C(3.2g,产率100%)。
LCMS m/z=167.1[M+1]。
第三步:4-[1-(环丙甲基)吡唑-3-羰基]-1-氧杂-4,9-二氮杂螺环[5.5]十一烷-9-羧酸叔丁酯(1D)
tert-butyl 4-[1-(cyclopropylmethyl)pyrazole-3-carbonyl]-1-oxa-4,9-diazaspiro[5.5]undecane-9-carboxylate
将1C(3.2g,20.0mmol)和1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-羧酸叔丁酯(5.3g,20.0mmol)加入DCM(50mL)中,加入三乙胺(13.0mL,100.0mmol)和1-丙基磷酸酐(17.0g,30.0mmol,50%乙酸乙酯溶液),室温搅拌3小时。滴加饱和碳酸氢钠水溶液(50mL),萃取,水相用DCM(100mL×1)萃取,合并后的有机相用无水硫酸钠干燥,过滤,滤液减压浓缩后,残留物用硅胶柱层析分离(DCM/甲醇(v/v)=15:1)得到黄色固体状的1D(2.4g,产率33%)。
1H NMR(400MHz,CDCl3)δ7.45-7.44(d,1H),6.69-6.68(d,1H),4.69-3.94(m,4H),3.75-3.55(m,7H),3.22-3.09(m,2H),1.80-1.76(m,2H),1.56-1.54(m,1H),1.42(s,9H),1.25-1.20(m,1H),0.63-0.59(m,2H),0.35-0.32(m,2H).
LCMS m/z=427.2[M+23]。
第四步:[1-(环丙基甲基)吡唑-3-基]-(1-氧杂-4,9-二氮杂[5.5]十一烷-4-基)甲酮(1E)
[1-(cyclopropylmethyl)pyrazol-3-yl]-(1-oxa-4,9-diazaspiro[5.5]undecan-4-yl)methanone
将1D(2.4g,5.9mmol)溶于DCM(30mL)中,加入三氟乙酸(6mL),室温搅拌2小时。反应液直接减压浓缩除去溶剂,向残余物中加入甲苯(5mL),减压浓缩后得到黄色液体状的1E的三氟乙酸盐(2.5g,产率92%)。
LCMS m/z=305.2[M+23]。
第五步:3-[2-[3-[2-[4-[1-(环丙基甲基)吡唑-3-羰基]-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-基]乙基]苯基]乙氧基]丙酸叔丁酯(1F)
tert-butyl 3-[2-[3-[2-[4-[1-(cyclopropylmethyl)pyrazole-3-carbonyl]-1-oxa-4,9-diazaspiro[5.5]undecan-9-yl]
ethyl]phenyl]ethoxy]propanoate
将1E的三氟乙酸盐(1.2g,3.0mmol)置于乙腈(25mL)中,加入中间体1(1.1g,3.0mmol)、碳酸钾(2.0g,15.0mmol)和水(0.5mL),60℃搅拌24小时。待反应冷至室温后,加入水(50mL),用乙酸乙酯(100mL×2)萃取,合并后的有机相用无水硫酸钠干燥,过滤,滤液减压浓缩后,残留物用硅胶柱层析分离(DCM/甲醇(v/v)=15:1)得到黄色液体状的1F(1.64g,产率96%)。
1H NMR(400MHz,CDCl3)δ7.47-7.46(m,1H),7.18-7.16(m,1H),7.04-7.01(m,3H),6.72-6.71(m,1H),4.13-3.97(m,3H),3.75-3.65(m,9H),2.87-2.45(m,13H),2.03-1.58(m,4H),1.48(s,9H),0.68-0.62(m,2H),0.38-0.34(m,2H).
LCMS m/z=581.4[M+1]。
第六步:3-[2-[3-[2-[4-[1-(环丙基甲基)吡唑-3-羰基]-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-基]乙基]苯基]乙氧基]丙酸(1G)
3-[2-[3-[2-[4-[1-(cyclopropylmethyl)pyrazole-3-carbonyl]-1-oxa-4,9-diazaspiro[5.5]undecan-9-yl]ethyl]phenyl]ethoxy]propanoic acid
将1F(1.64g,2.82mmol)溶于DCM(15mL)中,滴加三氟乙酸(5mL),室温搅拌4小时。反应液直接减压浓缩除去反应溶剂,向残余物中加入甲苯(10mL),减压浓缩后得到黄色液体状的1G的三氟乙酸盐(1.48g,产率100%)。
LCMS m/z=525.4[M+1]。
第七步:3-[2-[3-[2-[4-[1-(环丙基甲基)吡唑-3-羰基]-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-基]乙基]苯基]乙氧基]-N-(2,2-二甲氧基乙基)-N-甲基-丙酰胺(1H)
3-[2-[3-[2-[4-[1-(cyclopropylmethyl)pyrazole-3-carbonyl]-1-oxa-4,9-diazaspiro[5.5]undecan-9-yl]ethyl]phenyl]ethoxy]-N-(2,2-dimethoxyethyl)-N-methyl-propanamide
将1G的三氟乙酸盐(1.5g,2.8mmol)溶于DCM(20mL)中,加入甲氨基乙醛缩二甲醇(0.5g,4.2mmol)、三乙胺(1.43g,14.1mmol)和HATU(1.61g,4.2mmol),室温搅拌2小时。滴加饱和碳酸氢钠水溶液(50mL),用DCM(100mL×2)萃取,合并后的有机相用饱和食盐水(50mL)洗涤,无水硫酸钠干燥,过滤,滤液减压浓缩后,残留物用硅胶柱层析分离(DCM/甲醇(v/v)=15:1)得到黄色液体状的1H(1.3g,产率73.6%)。
LCMS m/z=626.5[M+1]。
第八步:N-[2-[[(2R)-2-[叔丁基(二甲基)硅基]氧基-2-(8-羟基-2-氧代-1H-喹啉-5-基)乙基]胺基]乙基-3-[2-[3-[2-[4-[1-(环丙基甲基)吡唑-3-羰基]-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-基]乙基]苯基]乙氧基]-N-(2,2-二甲氧基乙基)-N-甲基-丙酰胺(1I)
N-[2-[[(2R)-2-[tert-butyl(dimethyl)silyl]oxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]-3-[2-[3-[2-[4-[1-(cyclopropylmethyl)pyrazole-3-carbonyl]-1-oxa-4,9-diazaspiro[5.5]undecan-9-yl]ethyl]phenyl]ethoxy]-N-methyl-propanamide
将1H(0.63g,1.0mmol)溶于THF(10mL)中,加入TsOH·H2O(0.96g,5.0mmol),40℃搅拌1小时后,滴加饱和碳酸氢钠水溶液(50mL)淬灭反应,用DCM(100mL×2)萃取,合并后的有机相用无水硫
酸钠干燥,减压浓缩后得到的粗产品溶于DCM(10mL)和甲醇(3mL)的混合溶剂中,加入5-[(1R)-2-氨基-1-[叔丁基(二甲基)甲硅烷基]氧基甲基]-8-羟基-1H-喹啉-2-酮(0.34g,1.0mmol),室温搅拌30分钟后加入三乙酰氧基硼氢化钠(0.64g,3.0mmol),室温搅拌3小时。加入饱和碳酸氢钠水溶液(50mL),用DCM(100mL×2)萃取,合并后的有机相用无水硫酸钠干燥,过滤,滤液减压浓缩后,残留物用硅胶柱层析分离(DCM:甲醇(v/v)=1:0~8:1)得到黄色固体状的1I(0.50g,产率55%).
LCMS m/z=449.8[M/2+1]。
第九步:3-[2-[3-[2-[4-[1-(环丙甲基)吡唑-3-羰基]-1-氧杂-4,9-杂螺[5.5]十一烷-9-基]乙基]苯基]乙氧基]-N-[2-[[(2R)-2-羟基-2-(8-羟基-2-氧代-1H-喹啉-5-基)乙基]胺基]乙基]-N-甲基-丙酰胺(化合物1)
3-[2-[3-[2-[4-[1-(cyclopropylmethyl)pyrazole-3-carbonyl]-1-oxa-4,9-diazaspiro[5.5]undecan-9-yl]ethyl]phenyl]ethoxy]-N-[2-[[(2R)-2-hydroxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]-N-methyl-propanamide
将1I(0.50g,0.56mmol)溶于THF(10mL)中,加入三乙胺三氢氟酸(0.18g,1.1mmol),室温搅拌12小时。依次滴加饱和碳酸氢钠水溶液(50mL)和甲醇(5mL)淬灭反应,用DCM(100mL×2)萃取,合并后的有机相用无水硫酸钠干燥,过滤,滤液减压浓缩后,残留物用柱层析分离(DCM:甲醇(v/v)=1:0~8:1)得到黄色固体状的化合物1(0.35g,产率80%)。
1H NMR(400MHz,CDCl3)δ8.08(br,1H),7.47(s,1H),7.12-6.93(m,5H),6.69-6.67(m,2H),6.36(br,2H),5.48(br,1H),4.08-3.95(m,4H),3.62-3.47(m,10H),3.05-2.62(m,18H),1.93-1.84(m,5H),1.28-1.25(m,2H),0.63-0.61(m,2H),0.35-0.33(m,2H).
LCMS m/z=784.5[M+1]。
实施例2:3-[2-[3-[2-[4-[1-(环丙甲基)吡唑-3-羰基]-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-基]乙基]苯基]乙氧基]-N-[2-[[(2R)-2-羟基-2-(5-羟基-3-氧代-4H-1,4-苯并恶嗪-8-基)乙基]胺基]乙基]-N-甲基-丙酰胺(化合物2)
3-[2-[3-[2-[4-[1-(cyclopropylmethyl)pyrazole-3-carbonyl]-1-oxa-4,9-diazaspiro[5.5]undecan-9-yl]ethyl]phenyl]ethoxy]-N-[2-[[(2R)-2-hydroxy-2-(5-hydroxy-3-oxo-4H-1,4-benzoxazin-8-yl)ethyl]amino]ethyl]-N-methyl-propanamide
第一步:N-[2-[[(2R)-2-[叔丁基(二甲基)硅基]氧基-2-(5-羟基-3-氧代-4H-1,4-苯并恶嗪-8-基)乙基]胺基]乙基-3-[2-[3-[2-[4-[1-(环丙基甲基)吡唑-3-羰基]-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-基]乙基]苯基]乙氧基]-N-(2,2-二甲氧基乙基)-N-甲基-丙酰胺(2A)
N-[2-[[(2R)-2-[tert-butyl(dimethyl)silyl]oxy-2-(5-hydroxy-3-oxo-4H-1,4-benzoxazin-8-yl)ethyl]amino]ethyl]-3-[2-[3-[2-[4-[1-(cyclopropylmethyl)pyrazole-3-carbonyl]-1-oxa-4,9-diazaspiro[5.5]undecan-9-yl]ethyl]phenyl]ethoxy]-N-methyl-propanamide
将1H(0.63g,1.0mmol)溶于THF(10mL)中,加入TsOH·H2O(0.96g,5.0mmol),40℃搅拌1小时,滴加饱和碳酸氢钠水溶液(50mL)淬灭反应,用DCM(100mL×2)萃取,合并后的有机相用无水硫酸钠干燥,减压浓缩后得到的粗产品溶于DCM(10mL)和甲醇(3mL)的混合溶剂中,加入8-[(1R)-2-胺基-1-[叔丁基(二甲基)硅基]氧基-乙基]-5-羟基-4H-1,4-苯并恶嗪-3-酮(0.34g,1.0mmol),室温搅拌30分钟后加入三乙酰氧基硼氢化钠(0.64g,3.0mmol),室温搅拌3小时。加入饱和碳酸氢钠水溶液(50mL),用DCM(100mL×2)萃取,合并后的有机相用无水硫酸钠干燥,过滤,滤液减压浓缩后,残留物用硅胶柱层析分离(DCM:甲醇(v/v)=1:0~8:1)得到黄色固体状的2A(0.450g,产率50%)。
LCMS m/z=451.9[M/2+1]。
第二步:3-[2-[3-[2-[4-[1-(环丙甲基)吡唑-3-羰基]-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-基]乙基]苯基]乙氧基]-N-[2-[[(2R)-2-羟基-2-(5-羟基-3-氧代-4H-1,4-苯并恶嗪-8-基)乙基]胺基]乙基]-N-甲基-丙酰胺(化合物2)
3-[2-[3-[2-[4-[1-(cyclopropylmethyl)pyrazole-3-carbonyl]-1-oxa-4,9-diazaspiro[5.5]undecan-91-oxa-4,9-diazaspiro[5.5]undecan-9-yl]ethyl]phenyl]ethoxy]-N-[2-[[(2R)-2-hydroxy-2-(5-hydroxy-3-oxo-4H-1,4-benzoxazin-8-yl)ethyl]amino]ethyl]-N-methyl-propanamide
将2A(0.45g,0.50mmol)溶于THF(10mL)中,加入三乙胺三氢氟酸(0.16g,1.0mmol),室温搅拌12小时。依次滴加饱和碳酸氢钠水溶液(50mL)和甲醇(5mL)淬灭反应,用DCM(100mL×2)萃取,合并后的有机相用无水硫酸钠干燥,过滤,滤液减压浓缩后,残留物用硅胶柱层析分离(DCM:甲醇(v/v)=1:0~8:1)得到黄色固体状的化合物2(0.28g,产率71%)。
1H NMR(400MHz,CDCl3)δ7.48(s,1H),7.16-7.12(m,1H),6.97-6.87(m,4H),6.53-6.51(m,1H),6.51-6.45(m,1H),5.00-4.94(m,1H),4.48-4.42(m,2H),4.16-3.97(m,4H),3.65-3.42(m,10H),2.94-2.54(m,20H),1.95-1.77(m,4H),1.28-1.25(m,2H),0.63-0.61(m,2H),0.36-0.33(m,2H).
LCMS m/z=788.5[M+1]。
实施例3:N-[2-[[(2R)-2-羟基-2-(8-羟基-2-氧代-1H-喹啉-5-基)乙基]胺基]乙基]-N-甲基-3-[2-[3-[[4-(2-吡啶基)-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-基]甲基]苯基]乙氧基]丙酰胺(化合物3)
N-[2-[[(2R)-2-hydroxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]-N-methyl-3-[2-[3-[[4-(2-pyridyl)-1-oxa-4,9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]propanamide
第一步:4-(2-吡啶基)-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-羧酸叔丁酯(3B)
tert-butyl 4-(2-pyridyl)-1-oxa-4,9-diazaspiro[5.5]undecane-9-carboxylate
将1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-羧酸叔丁酯(5.12g,20.0mmol)加入1,4-二氧六环(50mL)中,加入2-氯吡啶(3A)(3.4g,30.0mmol),碳酸铯(13.0g,40.0mmol),4,5-双二苯基膦-9,9-二甲基氧杂蒽(1.6g,2.8mmol)和醋酸钯(0.3g,1.4mmol),氮气氛回流8小时。反应液冷却至室温,加入水(100mL),用乙酸乙酯(100mL×2)萃取,合并后的有机相用饱和食盐水(100mL×1)洗涤,无水硫酸钠干燥,过滤,滤液减压浓缩后,残留物用硅胶柱层析分离(石油醚/乙酸乙酯(v/v)=1:1)得到黄色液体状的3B(3.0g,产率45%)。
LCMS m/z=334.3[M+1]。
第二步:4-(2-吡啶基)-1-氧杂-4,9-二氮杂[5.5]十一烷(3C)
4-(2-pyridyl)-1-oxa-4,9-diazaspiro[5.5]undecane
将3B(0.5g,1.5mmol)溶于DCM(20mL)中。向反应瓶中加入三氟乙酸(4mL),室温搅拌2小时。减压浓缩除去反应溶剂,向残余物中加入甲苯(5mL),减压浓缩后得到黄色液体状的3C的三氟乙酸盐(0.53g,产率100%)。
第三步:3-[2-[3-[[4-(2-吡啶基)-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-基]甲基]苯基]乙氧基]丙酸叔丁酯(3D)
tert-butyl 3-[2-[3-[[4-(2-pyridyl)-1-oxa-4,9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]propanoate
将3C的三氟乙酸盐(0.53g,1.5mmol)置于乙腈(15mL)中,依次加入中间体2(0.53g,1.5mmol),碳酸钾(1.1g,7.5mmol)和水(0.5mL),室温搅拌8小时。加入水(50mL),用乙酸乙酯(100mL×2)萃取,合并后的有机相用无水硫酸钠干燥,过滤,滤液减压浓缩后,残留物用硅胶柱层析分离(DCM/甲醇(v/v)=15:1)得到黄色固体状的3D(0.50g,产率65%)。
LCMS m/z=496.4[M+1]。
第四步:3-[2-[3-[[4-(2-吡啶基)-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-基]甲基]苯基]乙氧基]丙酸(3E)
3-[2-[3-[[4-(2-pyridyl)-1-oxa-4,9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]propanoic acid
将3D(0.50g,1.0mmol)溶于DCM(15mL)中,滴加三氟乙酸(5mL),室温搅拌4小时。减压浓缩除去反应溶剂,向残余物中加入甲苯(10mL),减压浓缩后得到黄色液体状的3E的三氟乙酸盐(0.45g,产率100%)。
LCMS m/z=440.4[M+1]。
第五步:N-(2,2-二甲氧基乙基)-N-甲基-3-[2-[3-[[4-(2-吡啶基)-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-基]甲基]苯基]乙氧基]丙酰胺(3F)
N-(2,2-dimethoxyethyl)-N-methyl-3-[2-[3-[[4-(2-pyridyl)-1-oxa-4,9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]propanamide
将3E的三氟乙酸盐(0.45g,1.0mmol)溶于DCM(20mL)中,加入三乙胺(0.52g,5.0mmol),甲氨基乙醛缩二甲醇(0.18g,1.5mmol),HATU(0.58g,1.5mmol),室温搅拌2小时。滴加饱和碳酸氢钠水溶液(50mL),用DCM(100mL×2)萃取合并后的有机相用饱和食盐水(50mL)洗涤,无水硫酸钠干燥,过滤,滤液减压浓缩后,残留物用硅胶柱层析分离(DCM/甲醇(v/v)=15:1)得到黄色固体状的3F(0.50g,产率90.0%)。
LCMS m/z=541.4[M+1]。
第六步:N-[2-[[(2R)-2-[叔丁基(二甲基)硅基]氧基-2-(8-羟基-2-氧代-1H-喹啉-5-基)乙基]胺基]乙基-N-甲基-3-[2-[3-[[4-(2-吡啶基)-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-基]甲基]苯基]乙氧基]丙酰胺(3G)
N-[2-[[(2R)-2-[tert-butyl(dimethyl)silyl]oxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]-N-methyl-3-[2-[3-[[4-(2-pyridyl)-1-oxa-4,9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]propanamide
将3F(0.50g,0.92mmol)溶于THF(10mL)中,加入TsOH·H2O(0.88g,5.0mmol),40℃搅拌1小时后,滴加饱和碳酸氢钠水溶液(50mL)淬灭反应,用DCM(100mL×2)萃取,合并后的有机相用无水硫酸钠干燥,减压浓缩后得到的粗产品溶于DCM(10mL)和甲醇(3mL)的混合溶剂中,加入5-[(1R)-2-氨基-1-[叔丁基(二甲基)甲硅烷基]氧基甲基]-8-羟基-1H-喹啉-2-酮(0.31g,0.92mmol),室温搅拌30分钟后加入三乙酰氧基硼氢化钠(0.59g,2.8mmol),室温搅拌3小时。加入饱和碳酸氢钠水溶液(50mL),用DCM(100mL×2)萃取,合并后的有机相用无水硫酸钠干燥,过滤,滤液减压浓缩后,残留物用硅胶柱层析分离(DCM:甲醇(v/v)=1:0~8:1)得到黄色固体状的3G(0.33g,产率44%)
LCMS m/z=407.4[M/2+1]。
第七步:N-[2-[[(2R)-2-羟基-2-(8-羟基-2-氧代-1H-喹啉-5-基)乙基]胺基]乙基]-N-甲基-3-[2-[3-[[4-(2-吡啶基)-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-基]甲基]苯基]乙氧基]丙酰胺(化合物3)
N-[2-[[(2R)-2-hydroxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]-N-methyl-3-[2-[3-[[4-(2-pyridyl)-1-oxa-4,9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]propanamide
将3G(0.33g,0.41mmol)溶于THF(10mL)中,加入三乙胺三氢氟酸(0.65g,4.1mmol),室温搅拌12小时。依次滴加饱和碳酸氢钠水溶液(50mL)和甲醇(5mL)淬灭反应,用DCM(100mL×2)萃取,合并后的有机相用无水硫酸钠干燥,过滤,滤液减压浓缩后,残留物用硅胶柱层析分离(DCM:甲醇(v/v)=1:0~8:1)得到黄色固体状的化合物3(0.20g,产率71%)。
1H NMR(400MHz,CD3OD)δ8.32-8.29(d,1H),7.93(br,1H),7.76-7.74(m,1H),7.28-7.14(m,4H),7.07-7.05(d,1H),6.94-6.92(d,1H),6.81-6.78(m,1H),6.57-6.55(d,1H),5.30-5.27(m,1H),4.20(s,2H),3.69-3.67(m,2H),3.62-3.38(m,9H),3.22-3.03(m,13H),2.78-2.75(m,2H),2.57-2.56(m,2H),2.13-2.09(m,2H),1.75-1.72(m,2H).
LCMS m/z=699.5[M+1]。
实施例4:N-[2-[[(2R)-2-羟基-2-(8-羟基-2-氧代-1H-喹啉-5-基)乙基]胺基]乙基]-N-甲基-3-[2-[4-[2-[4-(2-吡啶基)-1-氧杂-4,9-杂螺二氮[5.5]十一烷-9-基]乙基]苯基]乙氧基]-丙酰胺(化合物4)
N-[2-[[(2R)-2-hydroxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]-N-methyl-3-[2-[4-[2-[4-(2-pyridyl)-1-oxa-4,9-diazaspiro[5.5]undecan-9-yl]ethyl]phenyl]ethoxy]propanamide
第一步:3-[2-[4-[2-[4-(2-吡啶基)-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-基]乙基]苯基]乙氧基]丙酸叔丁酯(4A)
tert-butyl 3-[2-[4-[2-[4-(2-pyridyl)-1-oxa-4,9-diazaspiro[5.5]undecan-9-yl]ethyl]phenyl]ethoxy]propanoate
将3C的三氟乙酸盐(0.5g,1.5mmol)加入乙腈(15mL)中,加入中间体3(0.55g,1.5mmol),碳酸钾(1.1g,7.5mmol)和水(0.5mL),60℃搅拌24小时。待反应冷至室温后,加入水(50mL),用乙酸乙酯(100mL×2)萃取,合并后的有机相用无水硫酸钠干燥,过滤,滤液减压浓缩后,残留物用硅胶柱层析分离(DCM/甲醇(v/v)=15:1)得到黄色液体状的4A(0.50g,产率64%)。
LCMS m/z=510.5[M+1]。
第二步:3-[2-[4-[2-[4-(2-吡啶基)-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-基]乙基]苯基]乙氧基]丙酸(4B)
3-[2-[4-[2-[4-(2-pyridyl)-1-oxa-4,9-diazaspiro[5.5]undecan-9-yl]ethyl]phenyl]ethoxy]propanoic acid
将4A(0.90g,1.8mmol)溶于DCM(15mL)中,滴加三氟乙酸(5mL),室温搅拌4小时。减压浓缩除去反应溶剂,向残余物中加入甲苯(10mL),减压浓缩后得到黄色液体状的4B的三氟乙酸盐(0.80g,产率100%)。
LCMS m/z=454.3[M+1]。
第三步:N-(2,2-二甲氧基乙基)-N-甲基-3-[2-[4-[2-[4-(2-吡啶基)-1-氧杂-4,9-二氮杂[5.5]十一烷-3-基]乙基]苯基]乙氧基]丙酰胺(4C)
N-(2,2-dimethoxyethyl)-N-methyl-3-[2-[4-[2-[4-(2-pyridyl)-1-oxa-4,9-diazaspiro[5.5]undecan-9-yl]ethyl]phenyl]ethoxy]propanamide
将4B的三氟乙酸盐(0.8g,1.8mmol)溶于DCM(20mL)中,依次加入三乙胺(0.89g,8.8mmol),甲氨基乙醛缩二甲醇(0.32g,2.6mmol)和HATU(1.0g,2.6mmol),室温搅拌2小时。滴加饱和碳酸氢钠水溶液(50mL),用DCM(100mL×2)萃取,合并后的有机相用饱和食盐水(50mL×1)洗涤,无水硫酸钠干燥,过滤,滤液减压浓缩后,残留物用硅胶柱层析分离(DCM/甲醇(v/v)=15:1)得到黄色液体状的4C(0.55g,产率56%)。
LCMS m/z=555.3[M+1]。
第四步:N-[2-[[(2R)-2-[叔丁基(二甲基)硅基]氧基-2-(8-羟基-2-氧代-1H-喹啉-5-基)乙基]胺基]乙基]-N-
甲基-3-[2-[4-[2-[4-(2-吡啶基)-1-氧杂-4,9-杂螺二氮[5.5]十一烷-9-基]乙基]苯基]乙氧基]丙酰胺(4D)
N-[2-[[(2R)-2-[tert-butyl(dimethyl)silyl]oxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]-N-methyl-3-[2-[4-[2-[4-(2-pyridyl)-1-oxa-4,9-diazaspiro[5.5]undecan-9-yl]ethyl]phenyl]ethoxy]propanamide
将4C(0.55g,0.99mmol)溶于THF(10mL)中,加入TsOH·H2O(0.94g,5.0mmol),40℃搅拌1小时后,滴加饱和碳酸氢钠水溶液(50mL)淬灭反应,用DCM(100mL×2)萃取,合并后的有机相用无水硫酸钠干燥,减压浓缩后得到的粗产品溶于DCM(10mL)和甲醇(3mL)的混合溶剂中,加入5-[(1R)-2-氨基-1-[叔丁基(二甲基)甲硅烷基]氧基甲基]-8-羟基-1H-喹啉-2-酮(0.34g,1.0mmol),室温搅拌30分钟后加入三乙酰氧基硼氢化钠(0.63g,3.0mmol),室温搅拌3小时。加入饱和碳酸氢钠水溶液(50mL),用DCM(100mL×2)萃取,合并后的有机相用无水硫酸钠干燥,过滤,滤液减压浓缩后,残留物用硅胶柱层析分离(DCM:甲醇(v/v)=1:0~8:1)得到黄色固体状的4D(0.34g,产率41%)。
第五步:N-[2-[[(2R)-2-羟基-2-(8-羟基-2-氧代-1H-喹啉-5-基)乙基]胺基]乙基]-N-甲基-3-[2-[4-[2-[4-(2-吡啶基)-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-基]乙基]苯基]乙氧基]丙酰胺(化合物4)
N-[2-[[(2R)-2-hydroxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]-N-methyl-3-[2-[4-[2-[4-(2-pyridyl)-1-oxa-4,9-diazaspiro[5.5]undecan-9-yl]ethyl]phenyl]ethoxy]propanamide
将4D(0.34g,0.41mmol)溶于THF(10mL)中,加入三乙胺三氢氟酸(1.98g,12.3mmol),室温搅拌12小时。向反应液中依次滴加饱和碳酸氢钠水溶液(50mL),甲醇(5mL)淬灭反应,用DCM(100mL×2)萃取,合并后的有机相用无水硫酸钠干燥,过滤,滤液减压浓缩后,残留物用硅胶柱层析分离(DCM:甲醇(v/v)=1:0~8:1)得到黄色固体状的化合物4(0.12g,产率41%)。
1H NMR(400MHz,DMSO-d6)δ10.22(br,1H)8.16-8.19(d,1H),8.08-8.10(dd,1H),7.49-7.53(m,1H),7.05-7.11(m,5H),6.90-6.92(d,1H),6.81-6.83(d,1H),6.60-6.63(dd,1H),5.74(s,1H),6.47-6.49(dd,1H),4.98-5.02(m,1H),3.69-3.71(m,2H),3.59-3.62(m,2H),3.52-3.57(m,2H),3.43-3.45(m,2H),3.37(s,2H),3.29-3.34(m,4H),2.92(s,2H)2.65-2.77(m,10H),2.32-2.37(m,3H),1.73-1.77(m,2H)1.52-1.57(m,2H)1.15-1.28(br,2H)。
LCMS m/z=713.5[M+1]。
实施例5:N-[2-[[(2R)-2-羟基-2-(8-羟基-2-氧代-1H-喹啉-5-基)乙基]胺基]乙基]-N-甲基-3-[2-[4-[[4-(2-丙炔基噻唑-4-羰基)-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-基]甲基]苯基]乙氧基]丙酰胺(化合物5)
N-[2-[[(2R)-2-hydroxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]-N-methyl-3-[2-[4-[[4-(2-prop-1-ynylthiazole-4-carbonyl)-1-oxa-4,9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]propanamide
第一步:2-丙炔基噻唑-4-甲酸乙酯(5B)
ethyl 2-prop-1-ynylthiazole-4-carboxylate
将2-溴噻唑-4-甲酸乙酯(5A)(11.8g,50.0mmol)溶于THF(200mL)中,依次加入三乙胺(7.6g,75.0mmol),三苯基膦(0.33g,1.25mmol),二三苯基膦二氯化钯(1.75g,2.5mmol),丙炔(120g,100.0mmol,3-4%庚烷溶液),60℃搅拌4小时。减压除去大部分反应溶剂,加入水(150mL),用乙酸乙酯萃取(200mL×2),合并后的有机相用无水硫酸钠干燥,过滤,滤液减压浓缩后,残留物用硅胶柱层析分离(石油醚/乙酸乙酯(v/v)=10:1)得到黄色固体状的5B(4.8g,产率49.2%)。
1H NMR(400MHz,CDCl3)δ8.06(s,1H),4.41-4.36(q,2H),2.07(s,3H),1.39-1.35(t,3H).
第二步:2-丙炔基噻唑-4-甲酸(5C)
2-prop-1-ynylthiazole-4-carboxylic acid
将5B(4.8g,24.6mmol)溶于THF(100mL)中,加入甲醇(10mL)、水(20mL)和氢氧化锂(1.8g,73.8mmol),室温搅拌2小时。减压除去大部分THF和甲醇,滴加1.0M稀盐酸水溶液调节pH到6,用乙酸乙酯萃取(150mL×2),合并后的有机相用无水硫酸钠干燥,减压浓缩后得到白色固体状的2-丙炔基噻唑-4-甲酸(5C)(4.1g,产率100%)。
第三步:4-(2-丙炔基噻唑-4-羰基)-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-羧酸叔丁酯(5D)
tert-butyl 4-(2-prop-1-ynylthiazole-4-carbonyl)-1-oxa-4,9-diazaspiro[5.5]undecane-9-carboxylate
将5C(4.2g,25.1mmol)和1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-羧酸叔丁酯(6.4g,25.1mmol)加入DCM(50mL)中,加入三乙胺(17.5mL,126.0mmol),1-丙基磷酸酐(24.0g,37.7mmol,50%乙酸乙酯溶液),室温搅拌3小时。滴加饱和碳酸氢钠水溶液(50mL),萃取分层后,水相用DCM(200mL×1)萃取,合并后的有机相用无水硫酸钠干燥,过滤,滤液减压浓缩后,残留物用硅胶柱层析分离(DCM/甲醇(v/v)=15:1)得到黄色液体状的5D(4.2g,产率41.2%)。
第四步:1-氧杂-4,9-二氮杂[5.5]十一烷-4-基-(2-丙炔基噻唑-4-基)甲酮(5E)
1-oxa-4,9-diazaspiro[5.5]undecan-4-yl-(2-prop-1-ynylthiazol-4-yl)methanone
称取5D(1.4g,3.5mmol)溶于DCM(20mL)中,加入三氟乙酸(4mL),室温搅拌2小时。减压浓缩除去反应溶剂,向残余物中加入甲苯(5mL),减压浓缩后得到黄色液体状的5E的三氟乙酸盐(1.5g,产率100%)。
第五步:3-[2-[4-[[4-(2-丙炔基噻唑-4-羰基)-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-基]甲基]苯基]乙氧基]丙酸叔丁酯(5F)
tert-butyl 3-[2-[4-[[4-(2-prop-1-ynylthiazole-4-carbonyl)-1-oxa-4,9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]propanoate
将5E的三氟乙酸盐(1.05g,2.9mmol)置于乙腈(15mL)中,加入中间体4(1.0g,2.9mmol)、碳酸钾(2.0g,15.0mmol)和水(0.5mL),室温搅拌4小时。加入水(50mL),用乙酸乙酯(100mL×2)萃取,合并后的有机相用无水硫酸钠干燥,过滤,滤液减压浓缩后,残留物用硅胶柱层析分离(DCM/甲醇(v/v)=15:1)得到黄色固体状的5F(1.0g,产率60%)。
LCMS m/z=568.3[M+1]。
第六步:3-[2-[4-[[4-(2-丙炔基噻唑-4-羰基)-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-基]甲基]苯基]乙氧基]丙酸(5G)
3-[2-[4-[[4-(2-prop-1-ynylthiazole-4-carbonyl)-1-oxa-4,9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]propanoic acid
将5F(1.0g,1.8mmol)溶于DCM(15mL)中,滴加三氟乙酸(5mL),室温搅拌4小时。减压浓缩除去反应溶剂,向残余物中加入甲苯(10mL),减压浓缩后得到黄色液体状的5G的三氟乙酸盐(0.9g,产率100%)。
LCMS m/z=512.3[M+1]。
第七步:N-(2,2-二甲氧基乙基)-N-甲基-3-[2-[4-[[4-(2-丙炔基噻唑-4-羰基)-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-基]甲基]苯基]乙氧基]丙酰胺(5H)
N-(2,2-dimethoxyethyl)-N-methyl-3-[2-[4-[[4-(2-prop-1-ynylthiazole-4-carbonyl)-1-oxa-4,9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]propanamide
将5G的三氟乙酸盐(1.0g,2.0mmol)溶于DCM(20mL)中,加入三乙胺(0.99g,10.0mmol),甲氨基乙醛缩二甲醇(0.35g,3.0mmol)和HATU(1.1g,3.0mmol),室温搅拌2小时。滴加饱和碳酸氢钠水溶液(50mL),用DCM(100mL×2)萃取,合并后的有机相用饱和食盐水(50mL×1)洗涤,无水硫酸钠干燥,过滤,滤液减压浓缩后,残留物用硅胶柱层析分离(DCM/甲醇(v/v)=15:1)得到黄色固体状的5H(1.0g,产率83.0%)。
LCMS m/z=613.4[M+1].
第八步:N-[2-[[(2R)-2-[叔丁基(二甲基)硅基]氧基-2-(8-羟基-2-氧代-1H-喹啉-5-基)乙基]胺基]乙-N-甲基-3-[2-[4-[[4-(2-丙炔基噻唑-4-羰基)-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-基]甲基]苯基]乙氧基]丙酰胺(5I)
N-[2-[[(2R)-2-[tert-butyl(dimethyl)silyl]oxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]-N-methyl-3-[2-[4-[[4-(2-prop-1-ynylthiazole-4-carbonyl)-1-oxa-4,9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]propanamide
将5H(1.0g,1.6mmol)溶于THF(10mL)中,加入TsOH·H2O(1.6g,8.2mmol),40℃搅拌1小时后,滴加饱和碳酸氢钠水溶液(50mL),用DCM(100m×2)萃取,合并后的有机相用无水硫酸钠干燥,减压浓缩后得到的粗产品溶于DCM(10mL)和甲醇(3mL)的混合溶剂中,加入5-[(1R)-2-氨基-1-[叔丁基(二甲基)甲硅烷基]氧基甲基]-8-羟基-1H-喹啉-2-酮(0.55g,1.6mmol),室温搅拌30分钟后加入三乙酰氧基硼
氢化钠(1.0g,4.9mmol),室温搅拌3小时。加入饱和碳酸氢钠水溶液(50mL)淬灭反应,用DCM(100mL×2)萃取,合并后的有机相用无水硫酸钠干燥,过滤,滤液减压浓缩后,残留物用硅胶柱层析分离(DCM:甲醇(v/v)=1:0~8:1)得到黄色固体状的5I(0.80g,产率55%).
LCMS m/z=886.5[M+1]。
第九步:N-[2-[[(2R)-2-羟基-2-(8-羟基-2-氧代-1H-喹啉-5-基)乙基]胺基]乙基]-N-甲基-3-[2-[4-[[4-(2-丙炔基噻唑-4-羰基)-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-基]甲基]苯基]乙氧基]丙酰胺(化合物5)
N-[2-[[(2R)-2-hydroxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]-N-methyl-3-[2-[4-[[4-(2-prop-1-ynylthiazole-4-carbonyl)-1-oxa-4,9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]propanamide
将5I(0.80g,0.90mmol)溶于THF(10mL)中,加入三乙胺三氢氟酸(1.5g,10.0mmol),室温搅拌12小时。依次滴加饱和碳酸氢钠水溶液(50mL)和甲醇(5mL)淬灭反应,用DCM(100mL×2)萃取,合并后的有机相用无水硫酸钠干燥,过滤,滤液减压浓缩后,残留物用硅胶柱层析分离(DCM:甲醇(v/v)=1:0~8:1)得到黄色固体状的化合物5(0.30g,产率43%)。
LCMS m/z=771.5[M+1]。
1H NMR(400MHz,CDCl3)δ7.91-7.88(m,1H),7.21-7.18(m,2H),7.09-6.96(m,3H),6.77-6.73(m,2H),6.34-6.30(m,1H),5.35-5.29(m,1H),3.92-3.65(m,14H),3.24-2.45(m,15H),2.11(s,3H),1.83-1.61(m,4H).
实施例6:N-[2-[[(2R)-2-羟基-2-(8-羟基-2-氧代-1H-喹啉-5-基)乙基]胺基]乙基]-N-甲基-3-[2-[3-[2-[4-(2-丙炔基噻唑-4-羰基)-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-基]乙基]苯基]乙氧基]丙酰胺(化合物6)
N-[2-[[(2R)-2-hydroxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]-N-methyl-3-[2-[3-[2-[4-(2-prop-1-ynylthiazole-4-carbonyl)-1-oxa-4,9-diazaspiro[5.5]undecan-9-yl]ethyl]phenyl]ethoxy]propanamide
第一步:3-[2-[3-[2-[4-(2-丙炔基噻唑-4-羰基)-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-基]乙基]苯基]乙氧基]丙酸叔丁酯(6A)
tert-butyl 3-[2-[3-[2-[4-(2-prop-1-ynylthiazole-4-carbonyl)-1-oxa-4,9-diazaspiro[5.5]undecan-9-yl]ethyl]phenyl]ethoxy]propanoate
将5E的三氟乙酸盐(0.50g,1.2mmol)加入乙腈(15mL)中,加入中间体1(0.44g,1.2mmol),碳酸钾(0.86g,6.0mmol),水(0.5mL)60℃搅拌24小时。加入水(50mL),用乙酸乙酯(100mL×2)萃取,合并后的有机相用无水硫酸钠干燥,过滤,滤液减压浓缩后,残留物用硅胶柱层析分离(DCM/甲醇(v/v)
=15:1)得到黄色固体状的6A(0.5g,产率70%)。
LCMS m/z=582.4[M+1]。
第二步:3-[2-[3-[2-[4-(2-丙炔基噻唑-4-羰基)-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-基]乙基]苯基]乙氧基]丙酸(6B)的三氟乙酸盐
3-[2-[3-[2-[4-(2-prop-1-ynylthiazole-4-carbonyl)-1-oxa-4,9-diazaspiro[5.5]undecan-9-yl]ethyl]phenyl]ethoxy]propanoic acid
将6A(0.5g,0.86mmol)溶于DCM(10mL)中,滴加三氟乙酸(4mL),室温搅拌4小时。减压浓缩除去反应溶剂,向残余物中加入甲苯(10mL),减压浓缩后得到黄色液体状的6B的三氟乙酸盐(0.45g,产率100%)。
第三步:N-(2,2-二甲氧基乙基)-N-甲基-3-[2-[3-[[4-(2-丙炔基噻唑-4-羰基)-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-基]乙基]苯基]乙氧基]丙酰胺(6C)
N-(2,2-dimethoxyethyl)-N-methyl-3-[2-[3-[2-[4-(2-prop-1-ynylthiazole-4-carbonyl)-1-oxa-4,9-diazaspiro[5.5]undecan-9-yl]ethyl]phenyl]ethoxy]propanamide
将6B的三氟乙酸盐(0.45g,0.86mmol)溶于DCM(15mL)中,依次加入三乙胺(0.43g,4.3mmol),甲氨基乙醛缩二甲醇(0.15g,1.3mmol),HATU(0.49g,1.3mmol),室温搅拌2小时。滴加饱和碳酸氢钠水溶液(50mL),用DCM(100mL×2)萃取,合并后的有机相用饱和食盐水(50mL×1)洗涤,无水硫酸钠干燥,过滤,滤液减压浓缩后,残留物用硅胶柱层析分离(DCM/甲醇(v/v)=15:1)得到黄色固体状的6C(0.50g,产率93.0%)。
LCMS m/z=627.4[M+1]。
第四步:N-[2-[[(2R)-2-[叔丁基(二甲基)硅基]氧-2-(8-羟基-2-氧代-1H-喹啉-5-基)乙基]胺基]乙基]-N-甲基-3-[2-[3-[[4-(2-丙炔基噻唑-4-羰基)-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-基]乙基]苯基]乙氧基]丙酰胺(6D)
N-[2-[[(2R)-2-[tert-butyl(dimethyl)silyl]oxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]-N-methyl-3-[2-[3-[2-[4-(2-prop-1-ynylthiazole-4-carbonyl)-1-oxa-4,9-diazaspiro[5.5]undecan-9-yl]ethyl]phenyl]ethoxy]propanamide
将6C(0.50g,0.80mmol)溶于THF(10mL)中,加入TsOH·H2O(0.76g,4.0mmol),40℃搅拌1小时,滴加饱和碳酸氢钠水溶液(50mL)淬灭反应,用DCM(100mL×2)萃取,合并后的有机相用无水硫酸钠干燥,减压浓缩后得到的粗产品溶于DCM(10mL)和甲醇(3mL)的混合溶剂中,加入5-[(1R)-2-氨基-1-[叔丁基(二甲基)甲硅烷基]氧基甲基]-8-羟基-1H-喹啉-2-酮(0.27g,0.80mmol),室温搅拌30分钟后加入三乙酰氧基硼氢化钠(0.51g,2.4mmol),室温搅拌3小时。加入饱和碳酸氢钠水溶液(50mL)淬灭反应,用DCM(100mL×2)萃取,合并后的有机相用无水硫酸钠干燥,过滤,滤液减压浓缩后,残留物用硅胶柱层析分离(DCM:甲醇(v/v)=1:0~8:1)得到黄色固体状的6D(0.16g,产率22%)
LCMS m/z=450.4[M/2+1]。
第五步:N-[2-[[(2R)-2-羟基-2-(8-羟基-2-氧代-1H-喹啉-5-基)乙基]胺基]乙基]-N-甲基-3-[2-[3-[2-[4-(2-丙炔基噻唑-4-羰基)-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-基]乙基]苯基]乙氧基]丙酰胺(化合物6)
N-[2-[[(2R)-2-hydroxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]-N-methyl-3-[2-[3-[2-[4-(2-prop-1-ynylthiazole-4-carbonyl)-1-oxa-4,9-diazaspiro[5.5]undecan-9-yl]ethyl]phenyl]ethoxy]propanamide
将6D(0.16g,0.18mmol)溶于THF(10mL)中,加入三乙胺三氢氟酸(0.29g,1.8mmol),室温搅拌12小时。依次滴加饱和碳酸氢钠水溶液(50mL)和甲醇(5mL)淬灭反应,用DCM(100mL×2)萃取,合并后的有机相用无水硫酸钠干燥,过滤,滤液减压浓缩后,残留物用硅胶柱层析分离(DCM:甲醇(v/v)=1:0~8:1)得到黄色固体状的化合物6(0.10g,产率72%)。
1H NMR(400MHz,CDCl3)δ8.08-8.02(m,1H),7.94-7.92(m,1H),7.15-7.08(m,2H),6.98-6.80(m,4H),6.56-6.48(m,1H),5.13-5.07(m,1H),3.95-3.43(m,10H),2.98-2.36(m,24H),2.20-1.67(m,4H)。
LCMS m/z=785.5[M+1]。
实施例7:3-[2-[3-[2-[4-(3-乙炔基苯甲酰基)-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-基]乙基]苯基]乙氧基]-N-[2-[[(2R)-2-羟基-2-(8-羟基-2-氧代-1H-喹啉-5-基)乙基]胺基]乙基]-N-甲基-丙酰胺(化合物7)
3-[2-[3-[2-[4-(3-ethynylbenzoyl)-1-oxa-4,9-diazaspiro[5.5]undecan-9-yl]ethyl]phenyl]ethoxy]-N-[2-[[(2R)-2-hydroxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]-N-methyl-propanamide
第一步:4-(3-乙炔基苯甲酰基)-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-羧酸叔丁酯(7B)
tert-butyl 4-(3-ethynylbenzoyl)-1-oxa-4,9-diazaspiro[5.5]undecane-9-carboxylate
将3-乙炔基苯甲酸(7A)(3.0g,20.0mmol)和1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-羧酸叔丁酯(5.2g,20.0mmol)置于DCM(100mL)中,依次加入三乙胺(14.0mL,100.0mmol),1-丙基磷酸酐(19.0g,30.0mmol,50%乙酸乙酯溶液),室温搅拌3小时。滴加饱和碳酸氢钠水溶液(100mL),萃取分层,水相用DCM(200mL×1)萃取,合并后的有机相用无水硫酸钠干燥,过滤,滤液减压浓缩后,残留物用硅胶柱层析分离(DCM/甲醇(v/v)=15:1)得到黄色液体状的7B(5.0g,产率64%)。
第二步:(3-乙炔基苯基)-(1-氧杂-4,9-二氮杂[5.5]十一烷-4-基)甲酮(7C)的三氟乙酸盐
(3-ethynylphenyl)-(1-oxa-4,9-diazaspiro[5.5]undecan-4-yl)methanone
将7B(0.6g,1.6mmol)溶于DCM(20mL),加入三氟乙酸(4mL),室温搅拌2小时。减压浓缩除去反应溶剂,向残余物中加入甲苯(5mL),减压浓缩后得到黄色液体状的7C的三氟乙酸盐(0.65g,产率
100%)。
LCMS m/z=285.3[M+1]。
第三步:3-[2-[3-[2-[4-(3-乙炔基苯甲酰基)-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-基]乙基]苯基]乙氧基]丙酸叔丁酯(7D)
tert-butyl 3-[2-[3-[2-[4-(3-ethynylbenzoyl)-1-oxa-4,9-diazaspiro[5.5]undecan-9-yl]ethyl]phenyl]ethoxy]propanoate
将7C的三氟乙酸盐(0.65g,1.6mmol)加入乙腈(15mL)中,加入中间体1(0.57g,1.6mmol),碳酸钾(1.1g,7.9mmol),水(0.5mL),60℃搅拌24小时。加入水(50mL),用乙酸乙酯(100mL×2),合并后的有机相用无水硫酸钠干燥,过滤,滤液减压浓缩后,残留物用硅胶柱层析分离(DCM/甲醇(v/v)=15:1)得到黄色固体状的7D(0.65g,产率73%)。
LCMS m/z=561.4[M+1]。
第四步:3-[2-[3-[2-[4-(3-乙炔基苯甲酰基)-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-基]乙基]苯基]乙氧基]丙酸(7E)
3-[2-[3-[2-[4-(3-ethynylbenzoyl)-1-oxa-4,9-diazaspiro[5.5]undecan-9-yl]ethyl]phenyl]ethoxy]propanoicacid
将7D(0.65g,1.2mmol)溶于DCM(15mL)中,滴加三氟乙酸(4mL),室温搅拌4小时。减压浓缩除去反应溶剂,向残余物中加入甲苯(10mL),减压浓缩后得到黄色液体状的7E的三氟乙酸盐(0.59g,产率100%)。
第五步:N-(2,2-二甲氧基乙基)-3-[2-[3-[2-[4-(3-乙炔基苯甲酰基)-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-基]乙基]苯基]乙氧基]-N-甲基-丙酰胺(7F)
N-(2,2-dimethoxyethyl)-3-[2-[3-[2-[4-(3-ethynylbenzoyl)-1-oxa-4,9-diazaspiro[5.5]undecan-9-yl]ethyl]phenyl]ethoxy]-N-methyl-propanamide
将7E的三氟乙酸盐(0.59g,1.2mmol)溶于DCM(15mL)中,依次加入三乙胺(0.81mL,6.0mmol),甲氨基乙醛缩二甲醇(0.21g,1.8mmol),HATU(0.67g,1.8mmol),室温搅拌2小时。滴加饱和碳酸氢钠水溶液(50mL),用DCM(100mL×2)萃取,合并后的有机相用饱和食盐水(50mL)洗涤,无水硫酸钠干燥,过滤,滤液减压浓缩后,残留物用硅胶柱层析分离(DCM/甲醇(v/v)=15:1)得到黄色固体状的7F(0.61g,产率86.0%)。
LCMS m/z=606.5[M+1]。
第六步:N-[2-[[(2R)-2-[叔丁基(二甲基)硅基]氧基-2-(8-羟基-2-氧代-1H-喹啉-5-基)乙基]胺基]乙基]-3-[2-[3-[2-[4-(3-乙炔基苯甲酰基)-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-基]乙基]苯基]乙氧基]-N-甲基-丙酰胺(7G)
N-[2-[[(2R)-2-[tert-butyl(dimethyl)silyl]oxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]-3-[2-[3-[2-[4-(3-ethynylbenzoyl)-1-oxa-4,9-diazaspiro[5.5]undecan-9-yl]ethyl]phenyl]ethoxy]-N-methyl-propanamide
将7F(0.61g,1.0mmol)溶于THF(10mL)中,加入TsOH·H2O(0.96g,5.0mmol),40℃搅拌1小时
后,滴加饱和碳酸氢钠水溶液(50mL)淬灭反应,用DCM(100mL×2)萃取,合并后的有机相用无水硫酸钠干燥,减压浓缩后得到的粗产品溶于DCM(10mL)和甲醇(3mL)的混合溶剂中,加入5-[(1R)-2-氨基-1-[叔丁基(二甲基)甲硅烷基]氧基甲基]-8-羟基-1H-喹啉-2-酮(0.34g,1.0mmol),室温搅拌30分钟后加入三乙酰氧基硼氢化钠(0.64g,3.0mmol),室温下搅拌3小时。加入饱和碳酸氢钠水溶液(50mL),用DCM(100mL×2)萃取,合并后的有机相用无水硫酸钠干燥,过滤,滤液减压浓缩后,残留物用硅胶柱层析分离(DCM:甲醇(v/v)=1:0~8:1)得到黄色固体状的7G(0.56g,产率63%)。
LCMS m/z=878.6[M+1]。
第七步:3-[2-[3-[2-[4-(3-乙炔基苯甲酰基)-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-基]乙基]苯基]乙氧基]-N-[2-[[(2R)-2-羟基-2-(8-羟基-2-氧代-1H-喹啉-5-基)乙基]胺基]乙基]-N-甲基-丙酰胺(化合物7)
3-[2-[3-[2-[4-(3-ethynylbenzoyl)-1-oxa-4,9-diazaspiro[5.5]undecan-9-yl]ethyl]phenyl]ethoxy]-N-[2-[[(2R)-2-hydroxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]-N-methyl-propanamide
将7G(0.56g,0.64mmol)溶于THF(10mL)中,加入三乙胺三氢氟酸(1.0g,6.4mmol),室温搅拌12小时。依次滴加饱和碳酸氢钠水溶液(50mL)和甲醇(5mL)淬灭反应,用DCM(100mL×2)萃取,合并后的有机相用无水硫酸钠干燥,过滤,滤液减压浓缩后,残留物用硅胶柱层析分离(DCM:甲醇(v/v)=1:0~8:1)得到黄色固体状的化合物7(0.20g,产率41%)。
1H NMR(400MHz,CDCl3)δ8.02-7.98(m,1H),7.56-7.48(m,2H),7.38-7.30(m,2H),7.16-7.12(m,1H),7.05-6.79(m,5H),6.51-6.42(m,1H),5.13-5.07(m,1H),3.72-3.43(m,16H),3.25-2.40(m,18H),1.89-1.56(m,2H)。
LCMS m/z=765.5[M+1]。
实施例8:3-[2-[3-[2-[4-(1,2-苯并噻唑-3-基)-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-基]乙基]苯基]乙氧基]-N-丁基-N-[2-[[(2R)-2-羟基-2-(8-羟基-2-氧代-1H-喹啉-5-基)乙基]胺基]乙基]丙酰胺(化合物8)
3-[2-[3-[2-[4-(1,2-benzothiazol-3-yl)-1-oxa-4,9-diazaspiro[5.5]undecan-9-yl]ethyl]phenyl]ethoxy]-N-butyl-N-[2-[[(2R)-2-hydroxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]propanamide
第一步:4-(1,2-苯并噻唑-3-基)-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-羧酸叔丁酯(8B)
tert-butyl 4-(1,2-benzothiazol-3-yl)-1-oxa-4,9-diazaspiro[5.5]undecane-9-carboxylate
将1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-羧酸叔丁酯(3.1g,12.1mmol)加入乙腈(20mL)中,加入3-氯-1,2-苯并异噻唑(8A)(2.3g,13.3mmol),1,5-二氮杂二环[5.4.0]十一-5-烯(DBU,1.8g,12.1mmol),50℃搅拌24小时。减压浓缩后除去大部分反应溶剂,硅胶柱层析分离(石油醚/乙酸乙酯(v/v)=4:1)得到黄色液体状的8B(2.5g,产率53%)。
1H NMR(400MHz,CDCl3)δ7.66-7.63(m,2H),7.59-7.55(m,1H),7.34-7.30(m,1H),3.79-3.76(m,4H),3.12-3.02(m,4H),2.86(s,2H),1.96-1.93(m,2H),1.48-1.41(m,11H)。
LCMS m/z=412.3[M+23]。
第二步:4-(1,2-苯并噻唑-3-基)-1-氧杂-4,9-二氮杂螺[5.5]十一烷(8C)
4-(1,2-benzothiazol-3-yl)-1-oxa-4,9-diazaspiro[5.5]undecane
将8B(0.78g,2.0mmol)溶于DCM(10mL)中,滴加三氟乙酸(4mL),室温搅拌4小时。减压浓缩除去反应溶剂,向残余物中加入甲苯(10mL),减压浓缩后得到黄色液体状的8C的三氟乙酸盐(0.84g,产率100%)。
第三步:3-[2-[3-[2-[4-(1,2-苯并噻唑-3-基)-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-基]乙基]苯基]乙氧基]丙酸叔丁酯(8D)
tert-butyl 3-[2-[3-[2-[4-(1,2-benzothiazol-3-yl)-1-oxa-4,9-diazaspiro[5.5]undecan-9-yl]ethyl]phenyl]ethoxy]propanoate
将8C的三氟乙酸盐(0.84g,2.0mmol)加入乙腈(15mL)中,依次加入中间体1(0.71g,2.0mmol),碳酸钾(1.38g,10.0mmol)和水(0.5mL),60℃搅拌24小时。加入水(50mL),用乙酸乙酯(100mL×2),合并后的有机相用无水硫酸钠干燥,过滤,滤液减压浓缩后,残留物用硅胶柱层析分离(DCM/甲醇(v/v)=15:1)得到黄色固体状的8D(0.85g,产率75%)。
1H NMR(400MHz,CDCl3)δ7.89-7.86(d,1H),7.83-7.81(d,1H),7.47-7.43(m,1H),7.37-7.33(m,1H),7.22-7.18(m,1H),7.06-7.05(m,3H),3.97-3.94(m,2H),3.72-3.68(m,4H),3.49-3.46(m,2H),3.33(s,2H),2.87-2.47(m,12H),2.18-2.14(m,2H),1.81-1.75(m,2H),1.44(s,9H)。第四步:3-[2-[3-[2-[4-(1,2-苯并噻唑-3-基)-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-基]乙基]苯基]乙氧基]丙酸(8E)
3-[2-[3-[2-[4-(1,2-benzothiazol-3-yl)-1-oxa-4,9-diazaspiro[5.5]undecan-9-yl]ethyl]phenyl]ethoxy]propanoicacid
将8D(1.0g,1.8mmol)溶于DCM(10mL)中,滴加三氟乙酸(4mL),室温搅拌4小时。减压浓缩除去反应溶剂,向残余物中加入甲苯(10mL),减压浓缩后得到黄色液体状的8E的三氟乙酸盐(0.9g,产率100%)。
LCMS m/z=510.3[M+1]。
第五步:3-[2-[3-[2-[4-(1,2-苯并噻唑-3-基)-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-基]乙基]苯基]乙氧基]-N-丁基-(2,2-二甲氧基乙基)丙酰胺(8F)
3-[2-[3-[2-[4-(1,2-benzothiazol-3-yl)-1-oxa-4,9-diazaspiro[5.5]undecan-9-yl]ethyl]phenyl]ethoxy]-N-butyl-N-(2,2-dimethoxyethyl)propanamide
将8E的三氟乙酸盐(0.90g,1.8mmol)溶于DCM(15mL)中,依次加入三乙胺(0.89g,8.8mmol),N-(2,2-二甲氧基乙基)丁基-1-胺(中间体5)(0.31g,1.9mmol),HATU(1.0g,2.6mmol),室温搅拌2小时。滴加饱和碳酸氢钠水溶液(50mL),用DCM(100mL×2)萃取,合并后的有机相用饱和食盐水(50mL×1)洗涤,无水硫酸钠干燥,过滤,滤液减压浓缩后,残留物用硅胶柱层析分离(DCM/甲醇(v/v)=15:1)得到黄色固体状的8F(1.2g,产率100%)。
LCMS m/z=653.5[M+1]。
第六步:3-[2-[3-[2-[4-(1,2-苯并噻唑-3-基)-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-基]乙基]苯基]乙氧基]-N-丁基-N-[2-[[(2R)-2-[叔丁基(二甲基)硅基]氧-2-(8-羟基-2-氧代-1H-喹啉-5-基)乙基]胺基]乙基]丙酰胺(8G)
3-[2-[3-[2-[4-(1,2-benzothiazol-3-yl)-1-oxa-4,9-diazaspiro[5.5]undecan-9-yl]ethyl]phenyl]ethoxy]-N-butyl-N-[2-[[(2R)-2-[tert-butyl(dimethyl)silyl]oxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]propanamide
将8F(0.65g,1.0mmol)溶于THF(10mL)中,加入TsOH·H2O(0.95g,5.0mmol),40℃搅拌1小时后,滴加饱和碳酸氢钠水溶液(50mL)淬灭反应,用DCM(100mL×2)萃取,合并后的有机相用无水硫酸钠干燥,减压浓缩后得到的粗产品溶于DCM(10mL)和甲醇(3mL)的混合溶剂中,加入5-[(1R)-2-氨基-1-[叔丁基(二甲基)甲硅烷基]氧基甲基]-8-羟基-1H-喹啉-2-酮(0.34g,1.0mmol),室温搅拌30分钟后加入三乙酰氧基硼氢化钠(0.63g,3.0mmol),室温搅拌3小时。加入饱和碳酸氢钠水溶液(50mL),用DCM(100mL×2)萃取,合并后的有机相用无水硫酸钠干燥,过滤,滤液减压浓缩后,残留物用硅胶柱层析分离(DCM:甲醇(v/v)=1:0~8:1)得到黄色固体状的8G(0.45g,产率49%)。
LCMS m/z=463.4[M/2+1]。
第七步:3-[2-[3-[2-[4-(1,2-苯并噻唑-3-基)-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-基]乙基]苯基]乙氧基]-N-丁基-N-[2-[[(2R)-2-羟基-2-(8-羟基-2-氧代-1H-喹啉-5-基)乙基]胺基]乙基]丙酰胺(化合物8)
3-[2-[3-[2-[4-(1,2-benzothiazol-3-yl)-1-oxa-4,9-diazaspiro[5.5]undecan-9-yl]ethyl]phenyl]ethoxy]-N-butyl-N-[2-[[(2R)-2-hydroxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]propanamide
将8G(0.45g,0.49mmol)溶于THF(10mL)中,加入三乙胺三氢氟酸(0.47g,2.9mmol),室温搅拌12小时。依次滴加饱和碳酸氢钠水溶液(50mL)和甲醇(5mL)淬灭反应,用DCM(100mL×2)萃取,合并后的有机相用无水硫酸钠干燥,过滤,滤液减压浓缩后,残留物用硅胶柱层析分离(DCM:甲醇(v/v)=1:0~8:1)得到黄色固体状的化合物8(0.2g,产率50%)。
1H NMR(400MHz,CDCl3)δ8.04(br,1H),7.84-7.79(m,2H),7.48-7.45(m,1H),7.33-7.30(m,1H),7.14-6.81(m,6H),6.49-6.45(m,1H),5.18-5.07(m,1H),3.92-3.91(m,2H),3.59-3.27(m,13H),2.81-2.54(m,17H),2.20-2.16(m,2H),1.84-1.81(m,2H),1.52-1.43(m,2H),0.92-0.86(m,3H)
LCMS m/z=811.5[M+1]。
实施例9:3-[2-[3-[2-[4-(1,2-苯并噻唑-3-基)-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-基]乙基]苯基]乙氧基]-N-丁基-N-[2-[[(2R)-2-羟基-2-(5-羟基-3-氧代-4H-1,4-苯并噁嗪-8-基)乙基]胺基]乙基]丙酰胺(化合物9)
3-[2-[3-[2-[4-(1,2-benzothiazol-3-yl)-1-oxa-4,9-diazaspiro[5.5]undecan-9-yl]ethyl]phenyl]ethoxy]-N-butyl-
N-[2-[[(2R)-2-hydroxy-2-(5-hydroxy-3-oxo-4H-1,4-benzoxazin-8-yl)ethyl]amino]ethyl]propanamide
第一步:3-[2-[3-[2-[4-(1,2-苯并噻唑-3-基)-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-基]乙基]苯基]乙氧基]-N-丁基-N-[2-[[(2R)-2-[叔丁基(二甲基)硅基]氧-2-(5-羟基-3-氧-4H-1,4-苯并噁嗪-8-基)乙基]胺基]乙基]丙酰胺(9A)
3-[2-[3-[2-[4-(1,2-benzothiazol-3-yl)-1-oxa-4,9-diazaspiro[5.5]undecan-9-yl]ethyl]phenyl]ethoxy]-N-butyl-N-[2-[[(2R)-2-[tert-butyl(dimethyl)silyl]oxy-2-(5-hydroxy-3-oxo-4H-1,4-benzoxazin-8-yl)ethyl]amino]ethyl]pro panamide
将8F(0.65g,1.0mmol)溶于THF(10mL)中,加入TsOH·H2O(0.95g,5.0mmol),40℃搅拌1小时后,滴加饱和碳酸氢钠水溶液(50mL)淬灭反应,用DCM(100mL×2)萃取,合并后的有机相用无水硫酸钠干燥,减压浓缩后得到的粗产品溶于DCM(10mL)和甲醇(3mL)的混合溶剂中,加入8-[(1R)-2-胺基-1-[叔丁基(二甲基)硅基]氧-乙基]-5-羟基-4氢-1,4-苯并噁嗪-3-酮(0.34g,1.0mmol),室温搅拌30分钟后加入三乙酰氧基硼氢化钠(0.63g,3.0mmol),室温搅拌3小时。加入饱和碳酸氢钠水溶液(50mL),用DCM(100mL×2)萃取,合并后的有机相用无水硫酸钠干燥,过滤,滤液减压浓缩后,残留物用硅胶柱层析分离(DCM:甲醇(v/v)=1:0~8:1)得到黄色固体状的9A(0.3g,产率32%)。
LCMS m/z=465.4[M/2+1]。
第二步:3-[2-[3-[2-[4-(1,2-苯并噻唑-3-基)-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-基]乙基]苯基]乙氧基]-N-丁基-N-[2-[[(2R)-2-羟基-2-(5-羟基-3-氧代-4H-1,4-苯并噁嗪-8-基)乙基]胺基]乙基]丙酰胺(化合物9)
3-[2-[3-[2-[4-(1,2-benzothiazol-3-yl)-1-oxa-4,9-diazaspiro[5.5]undecan-9-yl]ethyl]phenyl]ethoxy]-N-butyl-N-[2-[[(2R)-2-hydroxy-2-(5-hydroxy-3-oxo-4H-1,4-benzoxazin-8-yl)ethyl]amino]ethyl]propanamide
将9A(0.3g,0.32mmol)溶于THF(10mL)中,加入三乙胺三氢氟酸(0.29g,1.8mmol),室温搅拌12小时。向反应液中依次滴加饱和碳酸氢钠水溶液(50mL)和甲醇(5mL)淬灭反应,用DCM(100mL×2)萃取,合并后的有机相用无水硫酸钠干燥,过滤,滤液减压浓缩后,残留物用硅胶柱层析分离(DCM:甲醇(v/v)=1:0~8:1)得到黄色固体状的化合物9(0.12g,产率46%)。
1H NMR(400MHz,CDCl3)δ7.80-7.78(m,2H),7.48-7.45(m,1H),7.33-7.30(m,1H),7.14-6.77(m,5H),6.49-6.45(m,1H),5.12-5.07(m,1H),4.23-4.20(m,2H),3.89-3.60(m,28H),2.37-2.34(m,2H),2.04-1.98(m,2H),1.52-1.43(m,2H),1.29-1.26(m,4H),0.92-0.88(m,3H).
LCMS m/z=816.5[M+1]。
实施例10:N-[2-[[(2R)-2-羟基-2-(8-羟基-2-氧代-1H-喹啉-5-基)乙基]胺基]乙基]-N-甲基
-3-[2-[3-[2-[4-(2-吡啶基)-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-基]乙基]苯基]乙氧基]丙酰胺(化合物10)
N-[2-[[(2R)-2-hydroxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]-N-methyl-3-[2-[3-[2-[4-(2-pyridyl)-1-oxa-4,9-diazaspiro[5.5]undecan-9-yl]ethyl]phenyl]ethoxy]propanamide
第一步:3-[2-[3-[2-[4-(2-吡啶基)-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-基]乙基]苯基]乙氧基]丙酸叔丁酯(10A)
tert-butyl 3-[2-[3-[2-[4-(2-pyridyl)-1-oxa-4,9-diazaspiro[5.5]undecan-9-yl]ethyl]phenyl]ethox y]propanoate
将3C的三氟乙酸盐(0.46g,2.0mmol)加入乙腈(25mL)中,依次加入中间体1(0.70g,2.0mmol),碳酸钾(1.38g,10.0mmol),水(0.5mL),60℃搅拌24小时。待反应冷至室温后,加入水(50mL),用乙酸乙酯(100mL×2)萃取,合并后的有机相用无水硫酸钠干燥,过滤,滤液减压浓缩后,残留物用硅胶柱层析分离(DCM/甲醇(v/v)=15:1)得到黄色液体状的10A(0.70g,产率70%)。
LCMS m/z=510.3[M+1]。
第二步:3-[2-[3-[2-[4-(2-吡啶基)-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-基]乙基]苯基]-乙氧基]丙酸(10B)
3-[2-[3-[2-[4-(2-pyridyl)-1-oxa-4,9-diazaspiro[5.5]undecan-9-yl]ethyl]phenyl]ethoxy]propanoic acid
将10A(0.70g,1.40mmol)溶于DCM(15mL)中,滴加三氟乙酸(5mL),室温搅拌4小时。减压浓缩除去反应溶剂,向残余物中加入甲苯(10mL),减压浓缩后得到黄色液体状的10B的三氟乙酸盐(0.63g,产率100%)。
LCMS m/z=454.2[M+1]。
第三步:N-(2,2-二甲氧乙基)-N-甲基-3-[2-[3-[2-[4-(2-吡啶基)-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-基]乙基]苯基]乙氧基]丙酰胺(10C)
N-(2,2-dimethoxyethyl)-N-methyl-3-[2-[3-[2-[4-(2-pyridyl)-1-oxa-4,9-diazaspiro[5.5]undecan-9-yl]ethyl]phenyl]ethoxy]propanamide
将10B的三氟乙酸盐(0.63g,1.40mmol)溶于DCM(20mL)中,依次加入三乙胺(1.43g,7.0mmol),甲氨基乙醛缩二甲醇(0.37g,3.1mmol),HATU(0.79g,2.1mmol),室温搅拌2小时。滴加饱和碳酸氢钠水溶液(50mL),用DCM(100mL×2)萃取,合并后的有机相用饱和食盐水(50mL)洗涤,无水硫酸
钠干燥,过滤,滤液减压浓缩后,残留物用硅胶柱层析分离(DCM/甲醇(v/v)=15:1)得到黄色液体状的10C(0.75g,产率97.0%)。
LCMS m/z=555.5[M+1]。
第四步:N-[2-[[(2R)-2-(叔丁基(二甲基)硅基)氧-2-(8-羟基-2-氧-1H-喹啉-5-基)乙基]氨基]乙基]-N-甲基-3-[2-[3-[2-[4-(2-吡啶基)-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-基]乙基]苯基]乙氧基]丙酰胺(10D)
N-[2-[[(2R)-2-(tert-butyl(dimethyl)silyl)oxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]-N-methyl-3-[2-[3-[2-[4-(2-pyridyl)-1-oxa-4,9-diazaspiro[5.5]undecan-9-yl]ethyl]phenyl]ethoxy]propanamide
将10C(0.75g,1.4mmol)溶于THF(10mL)中,加入TsOH·H2O(1.3g,6.8mmol),40℃搅拌1小时后,滴加饱和碳酸氢钠水溶液(50mL)淬灭反应,用DCM(100mL×1)萃取,合并后的有机相用无水硫酸钠干燥,减压浓缩后得到的粗产品溶于DCM(10mL)和甲醇(3mL)的混合溶剂中,加入5-[(1R)-2-氨基-1-[叔丁基(二甲基)甲硅烷基]氧基甲基]-8-羟基-1H-喹啉-2-酮(0.45g,1.3mmol),室温搅拌30分钟后加入三乙酰氧基硼氢化钠(1.3g,6.1mmol),室温搅拌3小时。加入饱和碳酸氢钠水溶液(50mL),用DCM(100mL×2)萃取,合并后的有机相用无水硫酸钠干燥,过滤,滤液减压浓缩后,残留物用硅胶柱层析分离(DCM:甲醇(v/v)=1:0~8:1)得到黄色固体状的10D(0.38g,产率34%)。
LCMS m/z=827.6[M+1]。
第五步:N-[2-[[(2R)-2-羟基-2-(8-羟基-2-氧代-1H-喹啉-5-基)乙基]胺基]乙基]-N-甲基-3-[2-[3-[2-[4-(2-吡啶基)-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-基]乙基]苯基]乙氧基]丙酰胺(化合物10)
N-[2-[[(2R)-2-hydroxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]-N-methyl-3-[2-[3-[2-[4-(2-pyridyl)-1-oxa-4,9-diazaspiro[5.5]undecan-9-yl]ethyl]phenyl]ethoxy]propanamide
将10D(0.38g,0.46mmol)溶于THF(10mL)中,加入三乙胺三氢氟酸(0.15g,0.92mmol),室温搅拌12小时。依次滴加饱和碳酸氢钠水溶液(50mL),甲醇(5mL)淬灭反应,用DCM(100mL×2)萃取,合并后的有机相用无水硫酸钠干燥,过滤,滤液减压浓缩后,残留物用硅胶柱层析分离(DCM:甲醇(v/v)=1:0~8:1)得到黄色固体状的化合物10(0.100g,产率31%)。
1H NMR(400MHz,CDCl3)δ8.14(d,1H),8.00(s,1H),7.46(t,1H),7.14(t,1H),7.09–6.73(m,5H),6.61(t,3H),6.50–6.30(m,1H),5.17(d,1H),3.75(d,4H),3.51(dd,8H),3.14–2.40(m,18H),2.00(t,3H),1.74(t,2H).
LCMS m/z=357.4[M/2+1]。
实施例11:3-[2-[3-[2-[4-(4-乙基嘧啶基-2-基)-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-基]乙基]苯基]乙氧基]-N-[2-[[(2R)-2-羟基-2-(8-羟基-2-氧代-1H-喹啉-5-基)乙基]胺基]乙基]-N-甲基-丙酰胺(化合物11)
3-[2-[3-[2-[4-(4-ethylpyrimidin-2-yl)-1-oxa-4,9-diazaspiro[5.5]undecan-9-yl]ethyl]phenyl]ethoxy]-N-[2-[[(2R)-2-hydroxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]-N-methyl-propanamide
第一步:4-(4-乙基嘧啶基-2-基)-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-羧酸叔丁酯(11A)
tert-butyl 4-(4-ethylpyrimidin-2-yl)-1-oxa-4,9-diazaspiro[5.5]undecane-9-carboxylate
将1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-羧酸叔丁酯(0.512g,2.0mmol)加入DMF(20mL)中,依次加入2-氯-3-乙基嘧啶(0.285g,2.0mmol)、碳酸钾(0.552g,4.0mmol),50℃搅拌4小时。加入乙酸乙酯(200mL),水(100mL),萃取,水相用乙酸乙酯(100mL×1)萃取,合并后的有机相用饱和食盐水(100mL×1)洗涤,无水硫酸钠干燥,过滤,滤液减压浓缩后,残留物用硅胶柱层析分离(石油醚/乙酸乙酯(v/v)=1:1)得到黄色液体状的11A(0.667g,产率92.1%)。
LCMS m/z=363.3[M+1]。
第二步:2-(4-乙基嘧啶基-2-基)-1-氧杂-4,9-二氮杂螺[5.5]十一烷(11B)
4-(4-ethylpyrimidin-2-yl)-1-oxa-4,9-diazaspiro[5.5]undecane
将11A(0.80g,2.21mmol)溶于DCM(20mL)中,加入三氟乙酸(4mL),室温搅拌2小时。减压浓缩除去反应溶剂,向残余物中加入甲苯(5mL),减压浓缩后得到黄色液体状的11B的三氟乙酸盐(0.570g,产率98%)。
第三步:[2-[3-[[4-(4-乙基嘧啶基-2-基)-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-基]乙基]苯基]乙氧基]丙酸叔丁酯(11C)
tert-butyl 3-[2-[3-[[4-(4-ethylpyrimidin-2-yl)-1-oxa-4,9-diazaspiro[5.5]undecan-9-yl]ethyl]phenyl]ethoxy]propanoate
将11B的三氟乙酸盐(0.57g,2.17mmol)置于乙腈(25mL)中,依次加入中间体1(0.821g,2.30mmol),碳酸钾(1.50g,10.9mmol),水(0.5mL)。60℃搅拌24小时。待反应冷至室温后,加入水(50mL),用乙酸乙酯(100mL×2)萃取,合并后的有机相用无水硫酸钠干燥,减压浓缩后柱层析分离(DCM/甲醇(v/v)=15:1)得到黄色液体状的11C(1.00g,产率86%)。
LCMS m/z=539.5[M+1]。
第四步:3-[2-[3-[[4-(4-乙基嘧啶基-2-基)-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-基]乙基]苯基]乙氧基]丙酸(11D)
3-[2-[3-[[4-(4-ethylpyrimidin-2-yl)-1-oxa-4,9-diazaspiro[5.5]undecan-9-yl]ethyl]phenyl]ethoxy]propanoic
acid
将11C(1.00g,1.86mmol)溶于DCM(15mL)中,滴加三氟乙酸(5mL),室温搅拌4小时。减压浓缩除去反应溶剂,向残余物中加入甲苯(10mL),减压浓缩后得到黄色液体状的11D的三氟乙酸盐(0.90g,产率100%)。
LCMS m/z=483.3[M+1]。
第五步:N-(2,2-二甲氧乙基)-3-[2-[3-[[4-(4-乙基嘧啶基-2-基)-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-基]乙基]苯基]乙氧基]-N-甲基-丙酰胺(11E)
N-(2,2-dimethoxyethyl)-3-[2-[3-[[4-(4-ethylpyrimidin-2-yl)-1-oxa-4,9-diazaspiro[5.5]undecan-9-yl]ethyl]phenyl]ethoxy]-N-methyl-propanamide
将11D的三氟乙酸盐(0.90g,1.86mmol)溶于DCM(20mL)中,依次加入三乙胺(0.84g,9.3mmol),甲氨基乙醛缩二甲醇(0.266g,2.23mmol),HATU(1.07g,2.79mmol),室温搅拌2小时。滴加饱和碳酸氢钠水溶液(50mL),用DCM(100mL×2)萃取,合并后的有机相用饱和食盐水(50mL)洗涤,无水硫酸钠干燥,过滤,滤液减压浓缩后,残留物用硅胶柱层析分离(DCM/甲醇(v/v)=15:1)得到黄色液体状的11E(1.05g,产率96%)。
LCMS m/z=584.5[M+1]。
第六步:N-[2-[[(2R)-2-[叔丁基(二甲基)硅基]氧-2-(8-羟基-2-氧代-1H-喹啉-5-基)乙基]胺基]乙基-3-[2-[3-[[4-(4-乙基嘧啶基-2-基)-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-基]乙基]苯基]乙氧基]-N-甲基-丙酰胺(11F)
N-[2-[[(2R)-2-(tert-butyl(dimethyl)silyl)oxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]-N-methyl-3-[2-[3-[[4-(4-ethylpyrimidin-2-yl)-1-oxa-4,9-diazaspiro[5.5]undecan-9-yl]ethyl]phenyl]ethoxy]propanamide
将11E(1.02g,1.750mmol)溶于THF(10mL)中,加入TsOH·H2O(1.66g,8.75mmol),40℃搅拌1小时后,滴加饱和碳酸氢钠水溶液(50mL),用DCM(100mL×2)萃取,合并后的有机相用无水硫酸钠干燥,减压浓缩后得到的粗产品溶于DCM(10mL)和甲醇(3mL)的混合溶剂中,加入5-[(1R)-2-氨基-1-[叔丁基(二甲基)甲硅烷基]氧基甲基]-8-羟基-1H-喹啉-2-酮(0.586g,1.75mmol),室温搅拌30分钟后加入三乙酰氧基硼氢化钠(1.12g,5.25mmol),室温搅拌3小时。加入饱和碳酸氢钠水溶液(50mL)淬灭反应,用DCM(100mL×2)萃取,合并后的有机相用无水硫酸钠干燥,过滤,滤液减压浓缩后,残留物用硅胶柱层析分离(DCM:甲醇(v/v)=1:0~8:1)得到黄色固体状的11F(0.56g,产率37%)。
LCMS m/z=428.9[M/2+1]。
第七步:3-[2-[3-[2-[4-(4-乙基嘧啶基-2-基)-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-基]乙基]苯基]乙氧基]-N-[2-[[(2R)-2-羟基-2-(8-羟基-2-氧代-1H-喹啉-5-基)乙基]胺基]乙基]-N-甲基-丙酰(化合物11)
3-[2-[3-[2-[4-(4-ethylpyrimidin-2-yl)-1-oxa-4,9-diazaspiro[5.5]undecan-9-yl]ethyl]phenyl]ethoxy]-N-[2-[[(2R)-2-hydroxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]-N-methyl-propanamide
将11F(0.56g,0.65mmol)溶于THF(10mL)中,加入三乙胺三氢氟酸(0.21g,1.1mmol),室温搅拌12小时。依次滴加饱和碳酸氢钠水溶液(50mL),甲醇(5mL)淬灭反应,用DCM(100mL×2)萃取,合
并后的有机相用无水硫酸钠干燥,过滤,滤液减压浓缩后,残留物用硅胶柱层析分离(DCM:甲醇(v/v)=1:0~8:1)得到化合物11(0.16g,产率33%)。
1H NMR(400MHz,CDCl3)δ8.17(d,1H),8.09(d,1H),7.14(t,2H),6.97(dd,2H),6.89–6.79(m,1H),6.75(s,1H),6.61–6.48(m,1H),6.39(d,1H),5.14(d,1H),3.89–3.68(m,10H),3.66–3.52(m,5H),3.04(s,2H),2.94(d,3H),2.89–2.69(m,11H),2.65–2.55(m,5H),1.97(t,2H),1.81(s,2H),1.21(dt,4H).
LCMS m/z=742.5[M+1]。
实施例12:N-[2-[[(2R)-2-羟基-2-(8-羟基-2-氧代-1H-喹啉-5-基)乙基]胺基]乙基]-N-甲基-3-[2-[3-[2-[4-(4-甲基嘧啶基-2-基)-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-基]乙基]苯基]乙氧基]丙酰胺(化合物12)
N-[2-[[(2R)-2-hydroxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]-N-methyl-3-[2-[3-[2-[4-(4-methylpyrimidin-2-yl)-1-oxa-4,9-diazaspiro[5.5]undecan-9-yl]ethyl]phenyl]ethoxy]propanamide
第一步:4-(4-甲基嘧啶基-2-基)-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-羧酸叔丁酯(12A)
tert-butyl 4-(4-methylpyrimidin-2-yl)-1-oxa-4,9-diazaspiro[5.5]undecane-9-carboxylate
称取1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-羧酸叔丁酯(0.512g,2.0mmol)置于DMF(20mL)中,依次加入2-氯-3-甲基嘧啶(0.363g,2.0mmol)、碳酸钾(0.552g,4.0mmol),升温50℃搅拌4小时。向反应瓶中加入乙酸乙酯(200mL),水(100mL),萃取,水相用乙酸乙酯(100mL×1)萃取,合并后的有机相用饱和食盐水(100mL)洗涤,无水硫酸钠干燥,过滤,滤液减压浓缩后,残留物用硅胶柱层析分离(石油醚/乙酸乙酯(v/v)=1:1)得到黄色液体状的12A(0.615g,产率88.4%)。
LCMS m/z=349.3[M+1]。
第二步:4-(4-甲基嘧啶基-2-基)-1-氧杂-4,9-二氮杂螺[5.5]十一烷(12B)
4-(4-methylpyrimidin-2-yl)-1-oxa-4,9-diazaspiro[5.5]undecane
将12A(0.615g,1.77mmol),溶于DCM(20mL)中,加入三氟乙酸(4mL),室温搅拌2小时。减压浓缩除去反应溶剂,向残余物中加入甲苯(5mL),减压浓缩后得到黄色液体状的12B的三氟乙酸盐(0.44g,产率100%)。
LCMS m/z=249.3[M+1]。
第三步:3-[2-[3-[[4-(4-甲基嘧啶-2-基)-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-基]乙基]苯基]乙氧基]丙酸叔丁酯(12C)
tert-butyl 3-[2-[3-[[4-(4-methylpyrimidin-2-yl)-1-oxa-4,9-diazaspiro[5.5]undecan-9-yl]ethyl]phenyl]ethoxy]propanoate
将12B的三氟乙酸盐(0.44g,1.77mmol)置于乙腈(25mL)中,依次加入3-[2-[3-(2-溴乙基)苯基]乙氧基]丙酸叔丁酯(中间体1)(0.632g,1.77mmol),碳酸钾(1.221g,8.83mmol),水(0.5mL),60℃搅拌24小时。待反应冷至室温后,加入水(50mL),用乙酸乙酯(100mL×2)萃取,合并后的有机相用无水硫酸钠干燥,减压浓缩后柱层析分离(DCM/甲醇(v/v)=15:1)得到黄色液体状的12C(0.82g,产率88%)。
LCMS m/z=525.3[M+1]。
第四步:3-[2-[3-[[4-(4-甲基嘧啶基-2-基)-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-基]乙基]苯基]乙氧基]丙酸(12D)
3-[2-[3-[[4-(4-methylpyrimidin-2-yl)-1-oxa-4,9-diazaspiro[5.5]undecan-9-yl]ethyl]phenyl]ethoxy]propanoicacid
将12C(0.85g,1.62mmol)溶于DCM(15mL)中,滴加三氟乙酸(5mL),室温搅拌4小时。减压浓缩除去反应溶剂,向残余物中加入甲苯(10mL),减压浓缩后得到黄色液体状的12D的三氟乙酸盐(0.76g,产率100%)。
LCMS m/z=469.4[M+1]。
第五步:N-(2,2-二甲氧乙基)-N-甲基-3-[2-[3-[[4-(4-甲基嘧啶基-2-基)-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-基]乙基]苯基]乙氧基]-丙酰胺(12E)
N-(2,2-dimethoxyethyl)-N-methyl-3-[2-[3-[[4-(4-methylpyrimidin-2-yl)-1-oxa-4,9-diazaspiro[5.5]undecan-9-yl]ethyl]phenyl]ethoxy]propanamide
将12D的三氟乙酸盐(0.76g,1.62mmol)溶于DCM(20mL)中,依次加入三乙胺(0.82g,14.1mmol),甲氨基乙醛缩二甲醇(0.232g,1.95mmol),HATU(0.93g,2.43mmol),室温搅拌2小时。滴加饱和碳酸氢钠水溶液(50mL),用DCM(100mL×2)萃取,合并后的有机相用饱和食盐水(50mL×1)洗涤,无水硫酸钠干燥,过滤,滤液减压浓缩后,残留物用硅胶柱层析分离(DCM/甲醇(v/v)=15:1)得到黄色液体状的12E(0.9g,产率98%)。
LCMS m/z=570.3[M+1]。
第六步:N-[2-[[(2R)-2-[叔丁基(二甲基)硅基]氧-2-(8-羟基-2-氧代-1H-喹啉-5-基)乙基]胺基]乙基-N-甲基-3-[2-[3-[[4-(4-甲基嘧啶基-2-基)-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-基]乙基]苯基]乙氧基]丙酰胺(12F)
N-[2-[[(2R)-2-(tert-butyl(dimethyl)silyl)oxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]-N-methyl-3-[2-[3-[[4-(4-methylpyrimidin-2-yl)-1-oxa-4,9-diazaspiro[5.5]undecan-9-yl]ethyl]phenyl]ethoxy]propanamide
将12E(0.465g,0.82mmol)溶于THF(10mL)中,加入TsOH·H2O(0.79g,5.0mmol),40℃搅拌1小时后,滴加饱和碳酸氢钠水溶液(50mL)淬灭反应,用DCM(100mL×2)萃取,合并后的有机相用无水硫
酸钠干燥,减压浓缩后得到的粗产品溶于DCM(10mL)和甲醇(3mL)的混合溶剂中,加入5-[(1R)-2-氨基-1-[叔丁基(二甲基)甲硅烷基]氧基甲基]-8-羟基-1H-喹啉-2-酮(0.267g,0.80mmol),室温搅拌30分钟后加入三乙酰氧基硼氢化钠(0.53g,2.46mmol),室温搅拌3小时。加入饱和碳酸氢钠水溶液(50mL),用DCM(100mL×2)萃取,合并后的有机相用无水硫酸钠干燥,过滤,滤液减压浓缩后,残留物用硅胶柱层析分离(DCM:甲醇(v/v)=1:0~8:1)得到黄色固体状的12F(0.56g,产率81%)。
LCMS m/z=421.8[M/2+1]。
第七步:N-[2-[[(2R)-2-羟基-2-(8-羟基-2-氧代-1H-喹啉-5-基)乙基]胺基]乙基]-N-甲基-3-[2-[3-[2-[4-(4-甲基嘧啶基-2-基)-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-基]乙基]苯基]乙氧基]丙酰胺(化合物12)
N-[2-[[(2R)-2-hydroxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]-N-methyl-3-[2-[3-[2-[4-(4-methylpyrimidin-2-yl)-1-oxa-4,9-diazaspiro[5.5]undecan-9-yl]ethyl]phenyl]ethoxy]propanamide
将12F(0.56g,0.66mmol)溶于THF(10mL)中,加入三乙胺三氢氟酸(0.22g,1.1mmol),室温搅拌12小时。依次滴加饱和碳酸氢钠水溶液(50mL),甲醇(5mL)淬灭反应,用DCM(100mL×2)萃取,合并后的有机相用无水硫酸钠干燥,过滤,滤液减压浓缩后,残留物用硅胶柱层析分离(DCM:甲醇(v/v)=1:0~8:1)得到黄色固体状的化合物12(0.14g,产率29%)。
1H NMR(400MHz,CDCl3)δ8.13(d,1H),8.07(s,1H),7.13(dd,2H),6.98(dd,2H),6.92–6.77(m,2H),6.46(s,1H),6.38(d,1H),5.17(s,1H),3.80(d,2H),3.72(d,6H),3.65(s,1H),3.57(dd,2H),2.99–2.87(m,8H),2.86–2.52(m,10H),2.03(s,1H),1.97(t,2H),1.86(s,2H),1.32–1.26(m,7H).
LCMS m/z=728.5[M+1]。
实施例13:3-[2-[4-[2-[4-[1-(环丙甲基)吡唑-3-羰基]-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-基]乙基]苯基]乙氧基]-N-[2-[[(2R)-2-羟基-2-(8-羟基-2-氧代-1H-喹啉-5-基)乙基]胺基]乙基]-N-甲基-丙酰胺(化合物13)
3-[2-[4-[2-[4-[1-(cyclopropylmethyl)pyrazole-3-carbonyl]-1-oxa-4,9-diazaspiro[5.5]undecan-9-yl]ethyl]phenyl]ethoxy]-N-[2-[[(2R)-2-hydroxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]-N-methyl-propanamid
第一步:[1-(环丙甲基)吡唑-3-基]-[3-[2-[4-(2-羟基乙基)苯基]乙基]-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-基]甲酮(13A)
[1-(cyclopropylmethyl)pyrazol-3-yl]-[3-[2-[4-(2-hydroxyethyl)phenyl]ethyl]-1-oxa-4,9-diazaspiro[5.5]undecan-9-yl]methanone
将1E的三氟乙酸盐(0.80g,2.0mmol)置于乙腈(25mL)中,依次加入4-(2-溴乙基)苯乙醇(0.86g,3.8mmol),碳酸钾(1.38g,10.0mmol),水(0.5mL)。反应60℃搅拌3.5小时。待反应冷至室温后,依次加入水(50mL),用乙酸乙酯(100mL×2)萃取,合并后的有机相用无水硫酸钠干燥,过滤,滤液减压浓缩后,残留物用硅胶柱层析分离(DCM/甲醇(v/v)=15:1)得到黄色液体状的13A(0.82g,产率93%)。
第二步:3-[2-[4-[2-[8-[1-(环丙甲基)吡唑-3-羰基]-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-基]乙基]苯基]乙氧基]丙酸叔丁酯(13B)
tert-butyl3-[2-[4-[2-[8-[1-(cyclopropylmethyl)pyrazole-3-carbonyl]-1-oxa-4,9-diazaspiro[5.5]undecan-9-yl]ethyl]phenyl]ethoxy]propanoate
将13A(0.82g,1.8mmol)溶于乙腈(30mL)中,加入丙烯酸叔丁酯(1.0g,7.8mmol),苄基三甲基氢氧化铵(0.16g,0.96mmol,40%的甲醇溶液),室温搅拌3小时。减压浓缩除去大部分反应溶剂,柱层析分离(石油醚/乙酸乙酯(v/v)=10:1)得到黄色液体状的13B(1.01g,产率96%)。
第三步:3-[2-[4-[2-[4-[1-(环丙甲基)吡唑-3-羰基]-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-基]乙基]苯基]乙氧基]丙酸(13C)
3-[2-[4-[2-[4-[1-(cyclopropylmethyl)pyrazole-3-carbonyl]-1-oxa-4,9-diazaspiro[5.5]undecan-9-yl]ethyl]phenyl]ethoxy]propanoic acid
将13B(1.01g,1.74mmol)溶于DCM(15mL)中,滴加三氟乙酸(5mL),室温搅拌4小时。减压浓缩除去反应溶剂,向残余物中加入甲苯(10mL),减压浓缩后得到黄色液体状的13C的三氟乙酸盐(0.88g,产率96%)。
第四步:3-[2-[4-[2-[4-[1-(环丙甲基)吡唑-3-羰基]-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-基]乙基]苯基]乙氧基]-N-(2,2-二甲氧基乙基)-N-甲基-丙酰胺(13D)
3-[2-[4-[2-[4-[1-(cyclopropylmethyl)pyrazole-3-carbonyl]-1-oxa-4,9-diazaspiro[5.5]undecan-9-yl]ethyl]phenyl]ethoxy]-N-(2,2-dimethoxyethyl)-N-methyl-propanamide
将13C的三氟乙酸盐(0.88g,1.7mmol)溶于DCM(20mL)中,依次加入三乙胺(0.85g,8.4mmol),甲氨基乙醛缩二甲醇(0.3g,2.5mmol),HATU(0.96g,2.5mmol),室温搅拌2小时。滴加饱和碳酸氢钠水溶液(50mL),用DCM(100mL×2)萃取,合并后的有机相用饱和食盐水(50mL)洗涤,无水硫酸钠干燥,过滤,滤液减压浓缩后,残留物用硅胶柱层析分离(DCM/甲醇(v/v)=15:1)得到黄色液体状的13D(0.82g,产率78%)。
第五步:N-[2-[[(2R)-2-[叔丁基(二甲基)硅基]氧-2-(8-羟基-2-氧代-1H-喹啉-5-基)乙基]胺基]乙基-3-[2-[4-[2-[4-[1-(环丙基甲基)吡唑-3-羰基]-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-基]乙基]苯基]乙氧基]-N-(2,2-二甲氧基乙基)-N-甲基-丙酰胺(13E)
N-[2-[[(2R)-2-[tert-butyl(dimethyl)silyl]oxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]-3-[2-
[4-[2-[4-[1-(cyclopropylmethyl)pyrazole-3-carbonyl]-1-oxa-4,9-diazaspiro[5.5]undecan-9-yl]ethyl]phenyl]ethoxy]-N-methyl-propanamide
将13D(0.82g,1.3mmol)溶于THF(10mL)中,加入TsOH·H2O(1.2g,6.6mmol),40℃搅拌1小时后,滴加饱和碳酸氢钠水溶液(50mL)淬灭反应,用DCM(100mL×2)萃取,合并后的有机相用无水硫酸钠干燥,减压浓缩后得到的粗产品溶于DCM(10mL)和甲醇(3mL)的混合溶剂中,加入5-[(1R)-2-氨基-1-[叔丁基(二甲基)甲硅烷基]氧基甲基]-8-羟基-1H-喹啉-2-酮(0.34g,1.0mmol),室温搅拌30分钟后加入三乙酰氧基硼氢化钠(0.83g,3.9mmol),室温搅拌3小时。加入饱和碳酸氢钠水溶液(50mL),用DCM(100mL×2)萃取,合并后的有机相用无水硫酸钠干燥,过滤,滤液减压浓缩后,残留物用硅胶柱层析分离(DCM:甲醇(v/v)=1:0~8:1)得到黄色固体状的13E(0.42g,产率36%)
LCMS m/z=449.5[M/2+1]。
第六步:3-[2-[4-[2-[4-[1-(环丙甲基)吡唑-3-羰基]-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-基]乙基]苯基]乙氧基]-N-[2-[[(2R)-2-羟基-2-(8-羟基-2-氧代-1H-喹啉-5-基)乙基]胺基]乙基]-N-甲基-丙酰胺(化合物13)
3-[2-[4-[2-[4-[1-(cyclopropylmethyl)pyrazole-3-carbonyl]-1-oxa-4,9-diazaspiro[5.5]undecan-9-yl]ethyl]phenyl]ethoxy]-N-[2-[[(2R)-2-hydroxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]-N-methyl-propanamide
将13E(0.42g,0.47mmol)溶于THF(10mL)中,加入三乙胺三氢氟酸(1.98g,12.3mmol),室温搅拌12小时。依次滴加饱和碳酸氢钠水溶液(50mL),甲醇(5mL)淬灭反应,用DCM(100mL×2)萃取,合并后的有机相用无水硫酸钠干燥,过滤,滤液减压浓缩后,残留物用硅胶柱层析分离(DCM:甲醇(v/v)=1:0~8:1)得到黄色固体状的化合物13(0.09g,产率25%)。
1H NMR(400MHz,DMSO-d6)δ8.16-8.19(dd,1H),7.82(s,1H),7.06-7.09(m,5H),6.91-6.93(d,1H),6.57(s,1H),6.47-6.49(d,1H),5.74(s,1H),5.00-5.03(m,1H),4.00-4.02(d,2H),3.94(s,2H),3.51-3.58(m,12H),3.31-3.33(m,3H),2.92(s,1H),2.66-2.77(m,10H),2.36-2.44(m,5H),1.66(br,2H)1.52(br,2H),0.51-0.52(d,2H),0.34-0.38(m,2H)。
LCMS m/z=784.5[M+1]。
实施例14:N-丁基-N-[2-[[(2R)-2-羟基-2-(8-羟基-2-氧代-1H-喹啉-5-基)乙基]胺基]乙基]-3-[2-[3-[2-[4-(2-丙炔基噻唑-4-羰基)-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-基]乙基]苯基]乙氧基]丙酰胺(化合物14)
N-butyl-N-[2-[[(2R)-2-hydroxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]-3-[2-[3-[2-[4-(2-prop-1-ynylthiazole-4-carbonyl)-1-oxa-4,9-diazaspiro[5.5]undecan-9-yl]ethyl]phenyl]ethoxy]propanamide
第一步:N-丁基-N-(2,2-二甲氧基乙基)-3-[2-[3-[2-[4-(2-丙炔基噻唑-4-羰基)-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-基]乙基]苯基]乙氧基]丙酰胺(14A)
N-butyl-N-(2,2-dimethoxyethyl)-3-[2-[3-[2-[4-(2-prop-1-ynylthiazole-4-carbonyl)-1-oxa-4,9-diazaspiro[5.5]undecan-9-yl]ethyl]phenyl]ethoxy]propanamide
将6B的三氟乙酸盐(0.42g,0.80mmol)溶于DCM(15mL)中,依次加入N-(2,2-二甲氧基乙基)丁基-1-胺(中间体5)(0.13g,0.8mmol),三乙胺(0.40g,4.0mmol),HATU(0.46g,1.2mmol),室温搅拌2小时。滴加饱和碳酸氢钠水溶液(50mL),用DCM(100mL×2)萃取,合并后的有机相用饱和食盐水(50mL×1)洗涤,无水硫酸钠干燥,过滤,滤液减压浓缩后,残留物用硅胶柱层析分离(DCM/甲醇(v/v)=15:1)得到黄色固体状的14A(0.50g,产率94.0%)。
第二步:N-丁基-N-[2-[[(2R)-2-[叔丁基(二甲基)硅基]氧-2-(8-羟基-2-氧代-1H-喹啉-5-基)乙基]胺基]乙基]-3-[2-[3-[2-[4-(2-丙炔基噻唑-4-羰基)-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-基]乙基]苯基]乙氧基]丙酰胺(14B)
N-butyl-N-[2-[[(2R)-2-[tert-butyl(dimethyl)silyl]oxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]-3-[2-[3-[2-[4-(2-prop-1-ynylthiazole-4-carbonyl)-1-oxa-4,9-diazaspiro[5.5]undecan-9-yl]ethyl]phenyl]ethoxy]propanamide
将14A(0.50g,0.75mmol)溶于THF(10mL)中,加入TsOH·H2O(0.71g,3.7mmol),40℃搅拌1小时,滴加饱和碳酸氢钠水溶液(50mL)淬灭反应,用DCM(100mL×2)萃取,合并后的有机相用无水硫酸钠干燥,减压浓缩后得到的粗产品溶于DCM(10mL)和甲醇(3mL)的混合溶剂中,加入5-[(1R)-2-氨基-1-[叔丁基(二甲基)甲硅烷基]氧基甲基]-8-羟基-1H-喹啉-2-酮(0.25g,0.75mmol),室温搅拌30分钟后加入三乙酰氧基硼氢化钠(0.48g,2.2mmol),室温搅拌3小时。加入饱和碳酸氢钠水溶液(50mL),用DCM(100mL×2)萃取,合并后的有机相用无水硫酸钠干燥,过滤,滤液减压浓缩后,残留物用硅胶柱层析分离(DCM:甲醇(v/v)=1:0~8:1)得到黄色固体状的14B(0.30g,产率43%)。
LCMS m/z=471.5[M/2+1]。
第三步:N-丁基-N-[2-[[(2R)-2-羟基-2-(8-羟基-2-氧代-1H-喹啉-5-基)乙基]胺基]乙基]-3-[2-[3-[2-[4-(2-丙炔基噻唑-4-羰基)-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-基]乙基]苯基]乙氧基]丙酰胺(化合物14)
N-butyl-N-[2-[[(2R)-2-hydroxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]-3-[2-[3-[2-[4-(2-prop-1-ynylthiazole-4-carbonyl)-1-oxa-4,9-diazaspiro[5.5]undecan-9-yl]ethyl]phenyl]ethoxy]propanamide
将14B(0.30g,0.32mmol)溶于THF(10mL)中,加入三乙胺三氢氟酸(0.51g,3.2mmol),室温搅拌12小时。依次滴加饱和碳酸氢钠水溶液(50mL),甲醇(5mL)淬灭反应,用DCM(100mL×2)萃取,合并后的有机相用无水硫酸钠干燥,过滤,滤液减压浓缩后,残留物用硅胶柱层析分离(DCM:甲醇(v/v)=1:0~8:1)得到黄色固体状的化合物14(0.06g,产率20%)。
1H NMR(400MHz,CDCl3)δ8.05-7.91(m,2H),7.16-6.82(m,6H),6.51-6.43(m,1H),5.21-5.13(m,1H),
3.74-3.29(m,20H),2.76-2.47(m,14H),2.20-1.67(m,8H),0.92-0.87(m,3H)
LCMS m/z=828.4[M+1]。
实施例15:N-丁基-N-[2-[[(2R)-2-羟基-2-(8-羟基-2-氧代-1H-喹啉-5-基)乙基]胺基]乙基]-3-[2-[3-[2-[4-(1-甲基吡唑-3-基)-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-基]乙基]苯基]乙氧基]丙酰胺(化合物15)
N-butyl-N-[2-[[(2R)-2-hydroxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]-3-[2-[3-[2-[8-(1-methylpyrazol-3-yl)-1-oxa-4,9-diazaspiro[5.5]undecan-9-yl]ethyl]phenyl]ethoxy]propanamide
第一步:4-(1-甲基吡唑-3-基)-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-羧酸叔丁酯(15B)
tert-butyl 4-(1-methylpyrazol-3-yl)-1-oxa-4,9-diazaspiro[5.5]undecane-9-carboxylate
将1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-羧酸叔丁酯(2.6g,10.0mmol)置于1,4-二氧六环(20mL)中,依次加入3-氯-1-甲基-1H-吡唑(15A)(1.93g,12.0mmol),叔丁醇钠(2.4g,25.0mmol),醋酸钯(0.11g,0.5mmol),三叔丁基膦(2.0g,1.0mmol,10%的甲苯溶液),80℃搅拌12小时。待反应冷至室温,将反应液倒入饱和碳酸氢钠水溶液(30mL)中,用DCM萃取(100mL×2),合并后的有机相用无水硫酸钠干燥,过滤,滤液减压浓缩后,残留物用硅胶柱层析分离(石油醚/乙酸乙酯(v/v)=2:1)得到黄色液体状的15B(0.6g,产率18%)。
1H NMR(400MHz,CDCl3)δ7.15-7.14(d,1H),5.60-5.59(m,1H),3.83-3.81(m,4H),3.73(s,3H),3.20-3.11(m,4H),2.99(s,2H),1.98-1.95(m,2H),1.48-1.41(m,11H)。
第二步:4-(1-甲基吡唑-3-基)-1-氧杂-4,9-二氮杂螺[5.5]十一烷(15C)
4-(1-methylpyrazol-3-yl)-1-oxa-4,9-diazaspiro[5.5]undecane
将15B(0.60g,1.8mmol)溶于DCM(10mL)中,滴加三氟乙酸(4mL),室温搅拌4小时。减压浓缩除去反应溶剂,向残余物中加入甲苯(10mL),减压浓缩后得到黄色液体状的15C的三氟乙酸盐(0.65g,产率100%)。
第三步:3-[2-[3-[2-[4-(1-甲基吡唑-3-基)-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-基]乙基]苯基]乙氧基]丙
酸叔丁酯(15D)
tert-butyl 3-[2-[3-[2-[4-(1-methylpyrazol-3-yl)-1-oxa-4,9-diazaspiro[5.5]undecan-9-yl]ethyl]phenyl]ethoxy]propanoate
将15C的三氟乙酸盐(0.65g,1.8mmol)置于乙腈(15mL)中,加入中间体1(0.64,1.8mol),碳酸钾(1.2g,8.9mmol),水(0.5mL),60℃搅拌24小时。加入水(50mL),用乙酸乙酯(100mL×2)萃取,合并后的有机相用无水硫酸钠干燥,过滤,滤液减压浓缩后,残留物用硅胶柱层析分离(DCM/甲醇(v/v)=15:1)得到黄色固体状的15D(0.50g,产率55%)。
LCMS m/z=513.4[M+1]。
第四步:3-[2-[3-[2-[4-(1-甲基吡唑-3-基)-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-基]乙基]苯基]乙氧基]丙酸(15E)
3-[2-[3-[2-[4-(1-methylpyrazol-3-yl)-1-oxa-4,9-diazaspiro[5.5]undecan-9-yl]ethyl]phenyl]ethoxy]propanoicacid
将15D(0.5g,0.98mmol)溶于DCM(10mL)中,滴加三氟乙酸(4mL),搅拌4小时。减压浓缩除去反应溶剂,向残余物中加入甲苯(10mL),减压浓缩后得到黄色液体状的15E的三氟乙酸盐(0.45g,产率100%)。
第五步:N-丁基-N-(2,2-二甲氧基乙基)-3-[2-[3-[2-[4-(1-甲基吡唑-3-基)-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-基]乙基]苯基]乙氧基]丙酰胺(15F)
N-butyl-N-(2,2-dimethoxyethyl)-3-[2-[3-[2-[4-(1-methylpyrazol-3-yl)-1-oxa-4,9-diazaspiro[5.5]undecan-9-yl]ethyl]phenyl]ethoxy]propanamide
将15E的三氟乙酸盐(0.45g,1.0mmol)溶于DCM(15mL)中,依次加入三乙胺(0.5g,5.0mmol),N-(2,2-二甲氧基乙基)丁基-1-胺(中间体5)(0.16g,1.0mmol),HATU(0.56g,1.5mmol),室温搅拌2小时。滴加饱和碳酸氢钠水溶液(50mL),用DCM(100mL×2)萃取,合并后的有机相用饱和食盐水(50mL×1)洗涤,无水硫酸钠干燥,过滤,滤液减压浓缩后,残留物用硅胶柱层析分离(DCM/甲醇(v/v)=15:1)得到黄色固体状的15F(0.48g,产率81%)。
LCMS m/z=600.5[M+1]。
第六步:N-丁基-N-[2-[[(2R)-2-[叔丁基(二甲基)硅基]氧-2-(8-羟基-2-氧代-1H-喹啉-5-基)乙基]胺基]乙基]-3-[2-[3-[2-[4-(1-甲基吡唑-3-基)-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-基]乙基]苯基]乙氧基]-丙酰胺(15G)
N-butyl-N-[2-[[(2R)-2-[tert-butyl(dimethyl)silyl]oxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]-3-[2-[3-[2-[4-(1-methylpyrazol-3-yl)-1-oxa-4,9-diazaspiro[5.5]undecan-9-yl]ethyl]phenyl]ethoxy]propanamide
将15F(0.48g,0.8mmol)溶于THF(10mL)中,加入TsOH·H2O(0.76g,4.0mmol),40℃搅拌1小时后,滴加饱和碳酸氢钠水溶液(50mL)淬灭反应,用DCM(100mL×2)萃取,合并后的有机相用无水硫酸钠干燥,减压浓缩后得到的粗产品溶于DCM(10mL)和甲醇(3mL)的混合溶剂中,加入5-[(1R)-2-氨基-1-[叔丁基(二甲基)甲硅烷基]氧基甲基]-8-羟基-1H-喹啉-2-酮(0.27g,0.8mmol),室温搅拌30分钟后加入三乙酰氧基硼氢化钠(0.51g,2.4mmol),室温搅拌3小时。加入饱和碳酸氢钠水溶液(50mL),用DCM
(100mL×2)萃取,合并后的有机相用无水硫酸钠干燥,过滤,滤液减压浓缩后,残留物用硅胶柱层析分离(DCM:甲醇(v/v)=1:0~8:1)得到黄色固体状的15G(0.28g,产率40%).
LCMS m/z=436.9[M/2+1]。
第七步:N-丁基-N-[2-[[(2R)-2-羟基-2-(8-羟基-2-氧代-1H-喹啉-5-基)乙基]胺基]乙基]-3-[2-[3-[2-[4-(1-甲基吡唑-3-基)-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-基]乙基]苯基]乙氧基]丙酰胺(化合物15)
N-butyl-N-[2-[[(2R)-2-hydroxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]-3-[2-[3-[2-[8-(1-methylpyrazol-3-yl)-1-oxa-4,9-diazaspiro[5.5]undecan-9-yl]ethyl]phenyl]ethoxy]propanamide
将15G(0.28g,0.32mmol)溶于THF(10mL)中,加入三乙胺三氢氟酸(0.31g,1.9mmol),室温搅拌12小时。依次滴加饱和碳酸氢钠水溶液(50mL),甲醇(5mL)淬灭反应,用DCM(100mL×2)萃取,合并后的有机相用无水硫酸钠干燥,过滤,滤液减压浓缩后,残留物用硅胶柱层析分离(DCM:甲醇(v/v)=1:0~8:1)得到黄色固体状的化合物15(0.1g,产率40%)。
1H NMR(400MHz,CDCl3)δ8.09-8.07(m,1H),6.96-6.76(m,8H),6.54-6.45(m,1H),4.30-3.75(m,15H),3.30-2.59(m,21H),2.11-2.08(m,2H),1.90-1.86(m,2H),143-1.24(m,2H),0.92-0.86(m,3H)。
LCMS m/z=758.5[M+1]。
实施例16:3-[2-[3-[2-[4-(4-乙基-2-吡啶基)-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-基]乙基]苯基]乙氧基]-N-[2-[[(2R)-2-羟基-2-(8-羟基-2-氧代-1H-喹啉-5-基)乙基]胺基]乙基]-N-甲基-丙酰胺(化合物16)
3-[2-[3-[2-[4-(4-ethyl-2-pyridyl)-1-oxa-4,9-diazaspiro[5.5]undecan-9-yl]ethyl]phenyl]ethoxy]-N-[2-[[(2R)-2-hydroxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]-N-methyl-propanamide
第一步:2-氯-4-乙基-吡啶(16B)
2-chloro-4-ethyl-pyridine
将2-氨基-4-乙基吡啶(16A)(8.0g,65mmol)溶于100mL浓盐酸中,0℃,依次滴加亚硝酸钠(6.8g,98
mmol)的水溶液(40mL),氯化钠(7.7g,130mmol)的水溶液(40mL),滴加完毕后,继续搅拌30分钟。反应液用20%的氢氧化钠水溶液调节至碱性,用乙酸乙酯萃取(150mL×2),合并后的有机相用无水硫酸钠干燥,过滤,滤液减压浓缩后,残留物用硅胶柱层析分离(石油醚/乙酸乙酯(v/v)=15:1)得到黄色液体状的16B(6.5g,产率70%)。
1H NMR(400MHz,CDCl3)δ8.26-8.24(d,1H),7.16(s,1H),7.05-7.03(m,1H),2.67-2.61(m,2H),1.26-1.22(m,3H).
第二步:4-(4-乙基-2-吡啶基)-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-羧酸叔丁酯(16C)
tert-butyl 4-(4-ethyl-2-pyridyl)-1-oxa-4,9-diazaspiro[5.5]undecane-9-carboxylate
将1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-羧酸叔丁酯(5.12g,20.0mmol)溶于1,4-二氧六环(50mL)中,依次加16B(4.24g,30.0mmol),碳酸铯(13.0g,40.0mmol),4,5-双二苯基膦-9,9-二甲基氧杂蒽(1.6g,2.8mmol),醋酸钯(0.3g,1.4mmol),氮气氛回流反应8小时后。加入水(100mL),用乙酸乙酯(100mL×2)萃取,合并后的有机相用饱和食盐水洗(100mL),无水硫酸钠干燥,过滤,滤液减压浓缩后,残留物用硅胶柱层析分离(石油醚/乙酸乙酯(v/v)=1:1)得到黄色液体状的16C(3.0g,产率42%)。
LCMS m/z=362.3[M+1]。
第三步:4-(4-乙基-2-吡啶)-1-氧杂-4,9-二氮杂螺环[5.5]十一烷(16D)
4-(4-ethyl-2-pyridyl)-1-oxa-4,9-diazaspiro[5.5]undecane
将16C(0.5g,1.4mmol)溶于DCM(30mL)中,加入三氟乙酸(6mL),室温搅拌2小时。减压浓缩除去反应溶剂,向残余物中加入甲苯(5mL),减压浓缩后得到黄色液体状的16D的三氟乙酸盐(0.52g,产率100%)。
第四步:3-[2-[3-[2-[4-(4-乙基-2-吡啶基)-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-基]乙基]苯基]乙氧基]丙酸叔丁酯(16E)
tert-butyl 3-[2-[3-[2-[4-(4-ethyl-2-pyridyl)-1-oxa-4,9-diazaspiro[5.5]undecan-9-yl]ethyl]phenyl]ethoxy]propanoate
将16D的三氟乙酸盐(0.52g,1.4mmol)置于乙腈(15mL)中,依次加入中间体1(0.49g,1.4mmol),碳酸钾(0.95g,6.9mmol),水(0.5mL),60℃搅拌24小时。待反应冷至室温后,加入水(50mL),用乙酸乙酯(100mL×2)萃取,合并后的有机相用无水硫酸钠干燥,过滤,滤液减压浓缩后,残留物用硅胶柱层析分离(DCM/甲醇(v/v)=15:1)得到黄色液体状的16E(0.6g,产率80%)。
第五步:3-[2-[3-[2-[4-(4-乙基-2-吡啶基)-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-基]乙基]苯基]乙氧基]丙酸(16F)
3-[2-[3-[2-[4-(4-ethyl-2-pyridyl)-1-oxa-4,9-diazaspiro[5.5]undecan-9-yl]ethyl]phenyl]ethoxy]propanoicacid
将16E(0.78g,1.5mmol)溶于DCM(15mL)中,滴加三氟乙酸(5mL),室温搅拌4小时。减压浓缩除去反应溶剂,向残余物中加入甲苯(10mL),减压浓缩后得到黄色液体状的16F的三氟乙酸盐(0.7g,产率100%)。
LCMS m/z=482.4[M+1]。
第六步:N-(2,2-二甲氧基乙基)-3-[2-[3-[2-[4-(4-乙基-2-吡啶基)-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-基]乙基]苯基]乙氧基]-N-甲基-丙酰胺(16G)
N-(2,2-dimethoxyethyl)-3-[2-[3-[2-[4-(4-ethyl-2-pyridyl)-1-oxa-4,9-diazaspiro[5.5]undecan-9-yl]ethyl]phenyl]ethoxy]-N-methyl-propanamide
将16F的三氟乙酸盐(0.7g,1.5mmol)溶于DCM(20mL)中,依次加入三乙胺(0.74g,7.3mmol),甲氨基乙醛缩二甲醇(0.26g,2.2mmol),HATU(0.83g,2.2mmol),室温搅拌2小时。滴加饱和碳酸氢钠水溶液(50mL),用DCM(100mL×2)萃取,合并后的有机相用饱和食盐水(50mL×1)洗涤,无水硫酸钠干燥,过滤,滤液减压浓缩后,残留物用硅胶柱层析分离(DCM/甲醇(v/v)=15:1)得到黄色固体状的16G(0.55g,产率65%)。
第七步:N-[2-[[(2R)-2-[叔丁基(二甲基)硅基]氧-2-(8-羟基-2-氧代-1H-喹啉-5-基)乙基]胺基]乙基]-3-[2-[3-[2-[4-(4-乙基-2-吡啶基)-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-基]乙基]苯基]乙氧基]-N-甲基-丙酰胺(16H)
N-[2-[[(2R)-2-[tert-butyl(dimethyl)silyl]oxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]-3-[2-[3-[2-[4-(4-ethyl-2-pyridyl)-1-oxa-4,9-diazaspiro[5.5]undecan-9-yl]ethyl]phenyl]ethoxy]-N-methyl-propanamide
将16G(0.6g,1.0mmol)溶于THF(10mL)中,加入TsOH·H2O(0.96g,5.0mmol),40℃搅拌1小时后,滴加饱和碳酸氢钠水溶液(50mL)淬灭反应,用DCM(100mL×2)萃取,合并后的有机相用无水硫酸钠干燥,减压浓缩后得到的粗产品溶于DCM(10mL)和甲醇(3mL)的混合溶剂中,加入5-[(1R)-2-氨基-1-[叔丁基(二甲基)甲硅烷基]氧基甲基]-8-羟基-1H-喹啉-2-酮(0.34g,1.0mmol),室温搅拌30分钟后加入三乙酰氧基硼氢化钠(0.64g,3.0mmol),室温搅拌3小时。加入饱和碳酸氢钠水溶液(50mL),用DCM(100mL×2)萃取,合并后的有机相用无水硫酸钠干燥,过滤,滤液减压浓缩后,残留物用硅胶柱层析分离(DCM:甲醇(v/v)=1:0~8:1)得到黄色固体状的16H(0.55g,产率60%)。
LCMS m/z=428.4[M/2+1]。
第七步:3-[2-[3-[2-[4-(4-乙基-2-吡啶基)-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-基]乙基]苯基]乙氧基]-N-[2-[[(2R)-2-羟基-2-(8-羟基-2-氧代-1H-喹啉-5-基)乙基]胺基]乙基]-N-甲基-丙酰胺(化合物16)
3-[2-[3-[2-[4-(4-ethyl-2-pyridyl)-1-oxa-4,9-diazaspiro[5.5]undecan-9-yl]ethyl]phenyl]ethoxy]-N-[2-[[(2R)-2-hydroxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]-N-methyl-propanamide
将16H(0.55g,0.64mmol)溶于THF(10mL)中,加入三乙胺三氢氟酸(1.0g,6.4mmol),室温搅拌12小时。依次滴加饱和碳酸氢钠水溶液(50mL),甲醇(5mL)淬灭反应,用DCM(100mL×2)萃取,合并后的有机相用无水硫酸钠干燥,过滤,滤液减压浓缩后,残留物用硅胶柱层析分离(DCM:甲醇(v/v)=1:0~8:1)得到黄色固体状的化合物16(0.3g,产率60%)。
1H NMR(400MHz,CDCl3)δ8.10-8.05(m,2H),7.20-7.16(m,2H),6.97-6.94(m,2H),6.83-6.80(m,1H),6.70(s,1H),6.60-6.52(m,2H),6.47(s,1H),5.14-5.09(m,1H),3.81-3.37(m,13H),3.03-2.54(m,21H),2.05-1.98(m,2H),1.78(br,2H),1.28-1.20(m,3H).
LCMS m/z=371.4[M/2+1]。
实施例17:N-[2-[[(2R)-2-羟基-2-(8-羟基-2-氧代-1H-喹啉-5-基)乙基]胺基]乙基]-N-甲基-3-[2-[4-[[4-(2-
吡啶基-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-基]甲基]苯基]乙氧基]丙酰胺(化合物17)
N-[2-[[(2R)-2-hydroxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]-N-methyl-3-[2-[4-[[4-(2-pyridyl)-1-oxa-4,9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]propanamide
第一步:3-[2-[4-[[4-(2-吡啶基)-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-基]甲基]苯基]乙氧基]丙酸叔丁酯(17B)
tert-butyl 3-[2-[4-[[4-(2-pyridyl)-1-oxa-4,9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]propanoate
将17A(0.35g,1.50mmol)和3-[2-(4-醛基苯基)乙氧基]丙酸叔丁酯(4d)(0.42g,1.51mmol)溶于甲醇(16mL)溶剂中,室温搅拌1小时,加入三乙酰氧基硼氢化钠(0.95g,4.5mmol),室温搅拌3小时,加饱和碳酸氢钠溶液(10mL)淬灭反应,加入DCM(20mL),萃取分层,水相用DCM(50mL×2)萃取,合并有机相,用无水硫酸钠干燥,过滤,减压减压浓缩,残留物用硅胶柱层分离纯化得到黄色液体17B(0.342g,产率46%)。
LCMS m/z=496.3[M+1]。
第二步:3-[2-[4-[[4-(2-吡啶基)-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-基]甲基]苯基]乙氧基]丙酸(17C)
3-[2-[4-[[4-(2-pyridyl)-1-oxa-4,9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]propanoic acid
将17B(0.342g,0.69mmol)溶于DCM(15mL)中滴加三氟乙酸(5mL),室温搅拌4小时。减压浓缩除去反应溶剂,向残余物中加入甲苯(10mL),减压浓缩后得到黄色液体状的17C的三氟乙酸盐(0.309g,产率100%)。
LCMS m/z=440.4[M+1]。
第三步:N-(2,2-二甲氧乙基)-N-甲基-3-[2-[4-[[4-(2-吡啶基)-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-基]甲基]苯基]乙氧基]丙酰胺(17D)
N-(2,2-dimethoxyethyl)-N-methyl-3-[2-[4-[[4-(2-pyridyl)-1-oxa-4,9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]propanamide
将17C的三氟乙酸盐(0.303g,0.69mmol)溶于DCM(20mL)中,依次加入三乙胺(0.25g,3..45mmol),甲氨基乙醛缩二甲醇(0.098g,0.82mmol),HATU(0.393g,1.04mmol),室温搅拌2小时。滴加饱和碳酸氢钠水溶液(50mL),用DCM(100mL×2)萃取,合并后的有机相用饱和食盐水(50mL×1)洗涤,无水硫酸钠干燥,过滤,滤液减压浓缩后,残留物用硅胶柱层析分离(DCM/甲醇(v/v)=15:1)得到
黄色液体状的17D(0.37g,产率99.3%)。
LCMS m/z=541.3[M+1]。
第四步:N-[2-[[(2R)-2-[叔丁基(二甲基)硅基]氧-2-(8-羟基-2-氧代-1H-喹啉-5-基)乙基]胺基]乙基-N-甲基-3-[2-[4-[[4-(2-吡啶基)-1-氧-4,9-二氮杂螺[5.5]十一烷-9-基]甲基]苯基]乙氧基]丙酰胺(17E)
N-[2-[[(2R)-2-(tert-butyl(dimethyl)silyl)oxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]-N-methyl-3-[2-[4-[[4-(2-pyridyl)-1-oxa-4,9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]propanamide
将17D(0.372g,0.688mmol)溶于THF(10mL)中,加入TsOH·H2O(0.66g,3.44mmol),40℃搅拌1小时后,滴加饱和碳酸氢钠水溶液(50mL)淬灭反应,用DCM(100mL×2)萃取,合并后的有机相用无水硫酸钠干燥,减压浓缩后得到的残余物溶于DCM(10mL)和甲醇(3mL)的混合溶剂中,加入5-[(1R)-2-氨基-1-[叔丁基(二甲基)甲硅烷基]氧基甲基]-8-羟基-1H-喹啉-2-酮(0.23g,0.688mmol),室温搅拌30分钟后加入三乙酰氧基硼氢化钠(0.43g,2.0mmol),室温搅拌3小时后。加入饱和碳酸氢钠水溶液(50mL),用DCM(100mL×2)萃取,合并后的有机相用无水硫酸钠干燥,过滤,滤液减压浓缩后,残留物用硅胶柱层析分离(DCM:甲醇(v/v)=1:0~8:1)得到黄色固体状的17E(0.30g,产率553.6%)。
LCMS m/z=449.8[M/2+1]。
第五步:N-[2-[[(2R)-2-羟基-2-(8-羟基-2-氧代-1H-喹啉-5-基)乙基]胺基]乙基]-N-甲基-3-[2-[4-[[4-(2-吡啶基-1-氧杂-4,9--二氮杂螺[5.5]十一烷-9-基]甲基]苯基]乙氧基]丙酰胺(化合物17)
N-[2-[[(2R)-2-hydroxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]-N-methyl-3-[2-[4-[[4-(2-pyridyl)-1-oxa-4,9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]propanamide
将17E(0.30g,0.369mmol)溶于THF(10mL)中,加入三乙胺三氢氟酸(0.12g,0.738mmol),室温搅拌12小时。依次滴加饱和碳酸氢钠水溶液(50mL)和甲醇(5mL)淬灭反应,用DCM(100mL×2)萃取,合并后的有机相用无水硫酸钠干燥,过滤,滤液减压浓缩后,残留物用硅胶柱层析分离(DCM:甲醇(v/v)=1:0~8:1)得到黄色固体状的化合物17(0.080g,产率31%)。
1H NMR(400MHz,CDCl3)δ8.12(s,2H),7.49(d,1H),7.30(s,1H),7.20–6.93(m,3H),6.82(s,1H),6.60(s,3H),5.48(d,2H),3.76(s,7H),3.53(m,10H),3.35(s,3H),3.07(s,2H),3.03(d,1H),2.94(s,3H),2.80(s,2H),2.63(s,3H),2.01(d,4H),1.87(s,2H).
LCMS m/z=700.5[M+1]。
实施例18:3-[2-[3-[[4-(4-乙基-2-吡啶基)-1-氧杂-4,9-杂螺二氮[5.5]十一烷-9-基]甲基]苯基]乙氧基]-N-[2-[[(2R)-2-羟基-2-(8-羟基-2-氧代-1H-喹啉-5-基)乙基]胺基]乙基]-N-甲基-丙酰胺(化合物18)
3-[2-[3-[[4-(4-ethyl-2-pyridyl)-1-oxa-4,9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]-N-[2-[[(2R)-2-hydroxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]-N-methyl-propanamide
第一步:3-[2-[3-[[4-(4-乙基-2-吡啶基)-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-基]甲基]苯基]乙氧基]丙酸叔丁酯(18A)
tert-butyl 3-[2-[3-[[4-(4-ethyl-2-pyridyl)-1-oxa-4,9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]propanoate
将16D的三氟乙酸盐(0.52g,1.5mmol)置于乙腈(15mL)中,依次加入中间体2(0.53g,1.5mmol),碳酸钾(1.1g,7.7mmol),水(0.5mL),室温搅拌8小时。待反应冷至室温后,加入水(50mL),用乙酸乙酯(100mL×2)萃取,合并后的有机相用无水硫酸钠干燥,过滤,滤液减压浓缩后,残留物用硅胶柱层析分离(DCM/甲醇(v/v)=15:1)得到黄色液体状的18A(0.5g,产率65%)。
LCMS m/z=524.5[M+1].
第二步:3-[2-[3-[[4-(4-乙基-2-吡啶基)-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-基]甲基]苯基]乙氧基]丙酸(18B)
3-[2-[3-[[4-(4-ethyl-2-pyridyl)-1-oxa-4,9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]propanoicacid
将18A(0.5g,1.0mmol)溶于DCM(15mL)中,滴加三氟乙酸(5mL),室温搅拌4小时。减压浓缩除去反应溶剂,向残余物中加入甲苯(10mL),减压浓缩后得到黄色液体状的18B的三氟乙酸盐(0.45g,产率100%)。
LCMS m/z=468.4[M+1].
第三步:N-(2,2-二甲氧基乙基)-3-[2-[3-[[4-(4-乙基-2-吡啶基)-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-基]甲基]苯基]乙氧基]-N-甲基-丙酰胺(18C)
N-(2,2-dimethoxyethyl)-3-[2-[3-[[4-(4-ethyl-2-pyridyl)-1-oxa-4,9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]-N-methyl-propanamide
将18B的三氟乙酸盐(0.45g,1.0mmol)溶于DCM(20mL)中,依次加入三乙胺(0.50g,5.0mmol),甲氨基乙醛缩二甲醇(0.17g,1.4mmol),HATU(0.55g,1.4mmol),室温搅拌2小时。滴加饱和碳酸氢钠水溶液(50mL),用DCM(100mL×2)萃取,合并后的有机相用饱和食盐水(50mL×1)洗涤,无水硫酸钠干燥,过滤,滤液减压浓缩后,残留物用硅胶柱层析分离(DCM/甲醇(v/v)=15:1)得到黄色固体状的18C(0.50g,产率91%)。
LCMS m/z=570.5[M+1]。
第四步:N-[2-[[(2R)-2-[叔丁基(二甲基)硅基]氧-2-(8-羟基-2-氧代-1H-喹啉-5-基)乙基]胺基]乙
基]-3-[2-[3-[[4-(4-乙基-2-吡啶基)-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-基]甲基]苯基]乙氧基]-N-甲基-丙酰胺(18D)
N-[2-[[(2R)-2-[tert-butyl(dimethyl)silyl]oxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]-3-[2-[3-[[4-(4-ethyl-2-pyridyl)-1-oxa-4,9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]-N-methyl-propanamide
将18C(0.5g,0.88mmol)溶于THF(10mL)中,加入TsOH·H2O(0.84g,4.4mmol),40℃搅拌1小时后,滴加饱和碳酸氢钠水溶液(50mL)淬灭反应,用DCM(100mL×2)萃取,合并后的有机相用无水硫酸钠干燥,减压浓缩后得到的粗产品溶于DCM(10mL)和甲醇(3mL)的混合溶剂中,加入5-[(1R)-2-氨基-1-[叔丁基(二甲基)甲硅烷基]氧基甲基]-8-羟基-1H-喹啉-2-酮(0.29g,0.88mmol),室温搅拌30分钟后加入三乙酰氧基硼氢化钠(0.56g,2.6mmol),室温搅拌3小时。加入饱和碳酸氢钠水溶液(50mL),用DCM(100mL×2)萃取,合并后的有机相用无水硫酸钠干燥,过滤,滤液减压浓缩后,残留物用硅胶柱层析分离(DCM:甲醇(v/v)=1:0~8:1)得到黄色固体状的18D(0.55g,产率74%)。
LCMS m/z=422.15[M/2+1]。
第五步:3-[2-[3-[[4-(4-乙基-2-吡啶基)-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-基]甲基]苯基]乙氧基]-N-[2-[[(2R)-2-羟基-2-(8-羟基-2-氧代-1H-喹啉-5-基)乙基]胺基]乙基]-N-甲基-丙酰胺(化合物18)
3-[2-[3-[[4-(4-ethyl-2-pyridyl)-1-oxa-4,9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]-N-[2-[[(2R)-2-hydroxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]-N-methyl-propanamide
将18D(0.55g,0.65mmol)溶于THF(10mL)中,加入三乙胺三氢氟酸(1.0g,6.4mmol),室温搅拌12小时。依次滴加饱和碳酸氢钠水溶液(50mL)和甲醇(5mL)淬灭反应,用DCM(100mL×2)萃取,合并后的有机相用无水硫酸钠干燥,过滤,滤液减压浓缩后,残留物用硅胶柱层析分离(DCM:甲醇(v/v)=1:0~8:1)得到黄色固体状的化合物18(0.3g,产率60%)。
1H NMR(400MHz,CD3OD)δ8.44-8.41(d,1H),7.91-7.90(d,1H),7.39-7.33(m,5H),7.20(s,1H),7.06-7.04(d,1H),6.97-6.95(d,1H),6.69-6.66(d,1H),5.43-5.40(m,1H),4.32(s,2H),3.97-3.95(m,2H),3.75-3.62(m,11H),3.33-3.20(m,7H),3.11(s,3H),2.90-2.88(m,2H),2.78-2.67(m,4H),2.27-2.23(m,2H),1.93-1.89(m,2H),1.32-1.28(m,3H).
LCMS m/z=728.5[M+1]。
实施例19:N-[2-[[(2R)-2-羟基-2-(8-羟基-2-氧代-1H-喹啉-5-基)乙基]胺基]乙基]-N-甲基-3-[2-[3-[2-[4-(4-甲基嘧啶基-2-基)-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-基]甲基]苯基]乙氧基]丙酰胺(化合物19)
N-[2-[[(2R)-2-hydroxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]-N-methyl-3-[2-[3-[2-[4-(4-methylpyrimidin-2-yl)-1-oxa-4,9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]propanamide
第一步:叔丁基3-[2-[3-[[4-(4-甲基嘧啶-2-基)-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-基]甲基]苯基]乙氧基]丙酸酯(19B)
tert-butyl 3-[2-[3-[[4-(4-methylpyrimidin-2-yl)-1-oxa-4,9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]propanoate
将12B的三氟乙酸盐(0.50g,2.01mmol)置于乙腈(25mL)中,依次加入3-[2-[3-(氯甲基)苯基]乙氧基]丙酸叔丁酯(中间体6)(0.602g,2.01mmol),碳酸钾(1.39g,10.1mmol),水(0.5mL),60℃搅拌24小时。待反应冷至室温后,加入水(50mL),用乙酸乙酯(100mL×2)萃取,合并后的有机相用无水硫酸钠干燥,减压浓缩后柱层析分离(DCM/甲醇(v/v)=15:1)得到黄色液体状的19B(0.82g,产率88%)。
LCMS m/z=511.3[M+1]。
第二步:3-[2-[3-[[4-(4-甲基嘧啶基-2-基)-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-基]甲基]苯基]乙氧基]丙酸(19C)
3-[2-[3-[[4-(4-methylpyrimidin-2-yl)-1-oxa-4,9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]propanoic acid
将19B(1.0g,1.98mmol)溶于DCM(15mL)中滴加三氟乙酸(5mL),室温搅拌4小时。减压浓缩除去反应溶剂,向残余物中加入甲苯(10mL),减压浓缩后得到黄色液体状的19C的三氟乙酸盐(0.89g,产率100%)。
LCMS m/z=455.3[M+1].
第三步:N-(2,2-二甲氧乙基)-N-甲基-3-[2-[3-[[4-(4-甲基嘧啶基-2-基)-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-基]甲基]苯基]乙氧基]-丙酰胺(19D)
N-(2,2-dimethoxyethyl)-N-methyl-3-[2-[3-[[4-(4-methylpyrimidin-2-yl)-1-oxa-4,9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]propanamide
将19C的三氟乙酸盐(0.45g,0.98mmol)溶于DCM(20mL)中,依次加入三乙胺(0.99g,9.8mmol),甲氨基乙醛缩二甲醇(0.14g,1.2mmol),HATU(0.56g,1.5mmol),室温搅拌2小时。滴加饱和碳酸氢钠水溶液(50mL),用DCM(100mL×2)萃取,合并后的有机相用饱和食盐水(50mL)洗,无水硫酸钠干燥,减压浓缩后柱层析分离(DCM/甲醇(v/v)=15:1)得到黄色液体状的19D(0.54g,产率98%)。
LCMS m/z=556.3[M+1]。
第四步:N-[2-[[(2R)-2-[叔丁基(二甲基)硅基]氧基-2-(8-羟基-2-氧代-1H-喹啉-5-基)乙基]胺基]乙基-N-甲基-3-[2-[3-[[4-(4-甲基嘧啶基-2-基)-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-基]甲基]苯基]乙氧基]丙酰胺(19E)
N-[2-[[(2R)-2-(tert-butyl(dimethyl)silyl)oxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]-N-methyl-3-[2-[3-[[4-(4-methylpyrimidin-2-yl)-1-oxa-4,9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]propanamide
将19D(0.540g,0.97mmol)溶于THF(10mL)中,加入TsOH·H2O(0.95g,5.0mmol),40℃搅拌1小时,滴加饱和碳酸氢钠水溶液(50mL)淬灭反应,用DCM(100mL×2)萃取,合并后的有机相用无水硫酸钠干燥,减压浓缩后得到的粗产品溶于DCM(10mL)和甲醇(3mL)的混合溶剂中,加入5-[(1R)-2-氨基-1-[叔丁基(二甲基)甲硅烷基]氧基甲基]-8-羟基-1H-喹啉-2-酮(0.325g,0.97mmol),室温搅拌30分钟后加入三乙酰氧基硼氢化钠(0.62g,2.91mmol),室温搅拌3小时。加入饱和碳酸氢钠水溶液(50mL),用DCM(100mL×2)萃取,合并后的有机相用无水硫酸钠干燥,减压浓缩后柱层析分离(DCM:甲醇(v/v)=1:0~8:1)得到黄色固体状的19E(0.40g,产率50%)。
LCMS m/z=829.5[M+1]。
第五步:N-[2-[[(2R)-2-羟基-2-(8-羟基-2-氧代-1H-喹啉-5-基)乙基]胺基]乙基]-N-甲基-3-[2-[3-[2-[4-(4-甲基嘧啶基-2-基)-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-基]甲基]苯基]乙氧基]丙酰胺(化合物19)
N-[2-[[(2R)-2-hydroxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]-N-methyl-3-[2-[3-[2-[4-(4-methylpyrimidin-2-yl)-1-oxa-4,9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]propanamide
将19E(0.40g,0.48mmol)溶于THF(10mL)中,加入三乙胺三氢氟酸(0.14g,0.72mmol),室温搅拌12小时。依次滴加饱和碳酸氢钠水溶液(50mL),甲醇(5mL)淬灭反应,用DCM(100mL×2)萃取,合并后的有机相用无水硫酸钠干燥,减压浓缩后柱层析分离(DCM:甲醇(v/v)=1:0~8:1)得到黄色固体状的化合物19(0.11g,产率59%)。
1H NMR(400MHz,CD3OD)δ8.40(d,1H),8.19(d,1H),7.42–7.24(m,5H),7.03(d,1H),6.75–6.54(m,2H),5.40(d,1H),4.86(dd,14H),3.87–3.64(m,12H),2.89(t,2H),2.67(t,2H),2.39(s,3H),2.19(d,2H),1.79(s,2H).
LCMS m/z=715.5[M+1]。
实施例20:N-[2-[[(2R)-2-羟基-2-(8-羟基-2-氧代-1H-喹啉-5-基)乙基]胺基]乙基]-N-乙基-3-[2-[3-[2-[4-(4-甲基嘧啶基-2-基)-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-基]乙基]苯基]乙氧基]丙酰胺(化合物20)
N-[2-[[(2R)-2-hydroxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]-N-ethyl-3-[2-[3-[2-[4-(4-methylpyrimidin-2-yl)-1-oxa-4,9-diazaspiro[5.5]undecan-9-yl]ethyl]phenyl]ethoxy]propanamide
第一步:N-(2,2-二甲氧乙基)-N-乙基-3-[2-[3-[[4-(4-甲基嘧啶基-2-基)-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-基]乙基]苯基]乙氧基]-丙酰胺(20B)
N-(2,2-dimethoxyethyl)-N-ethyl-3-[2-[3-[[4-(4-methylpyrimidin-2-yl)-1-oxa-4,9-diazaspiro[5.5]undecan-9-yl]ethyl]phenyl]ethoxy]propanamide
将12D的三氟乙酸盐(0.525g,1.12mmol)溶于DCM(20mL)中,依次加入三乙胺(0.34g,3.36mmol),乙氨基乙醛缩二甲醇(0.179g,1.34mmol),HATU(0.852g,2.24mmol),室温搅拌2小时。滴加饱和碳酸氢钠水溶液(50mL),用DCM(100mL×2)萃取,合并后的有机相用饱和食盐水(50mL)洗涤,无水硫酸钠干燥,减压浓缩后柱层析分离(DCM/甲醇(v/v)=15:1)得到黄色液体状的20B(0.65g,产率98%)。
LCMS m/z=584.4[M+1].
第二步:N-[2-[[(2R)-2-[叔丁基(二甲基)硅基]氧基-2-(8-羟基-2-氧代-1H-喹啉-5-基)乙基]胺基]乙基-N-乙基-3-[2-[3-[[4-(4-甲基嘧啶基-2-基)-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-基]乙基]苯基]乙氧基]丙酰胺(20C)
N-[2-[[(2R)-2-(tert-butyl(dimethyl)silyl)oxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]-N-ethyl-3-[2-[3-[[4-(4-methylpyrimidin-2-yl)-1-oxa-4,9-diazaspiro[5.5]undecan-9-yl]ethyl]phenyl]ethoxy]propanamide
将20B(0.65g,1.12mmol)溶于THF(10mL)中,加入TsOH·H2O(1.09g,5.0mmol),40℃搅拌1小时后,滴加饱和碳酸氢钠水溶液(50mL)淬灭反应,用DCM(100mL×2)萃取,合并后的有机相用无水硫酸钠干燥,减压浓缩后得到的粗产品溶于DCM(10mL)和甲醇(3mL)的混合溶剂中,加入5-[(1R)-2-氨基-1-[叔丁基(二甲基)甲硅烷基]氧基甲基]-8-羟基-1H-喹啉-2-酮(0.375g,1.12mmol),室温搅拌30分钟后加入三乙酰氧基硼氢化钠(0.712g,3.36mmol),室温搅拌3小时。加入饱和碳酸氢钠水溶液(50mL),用DCM(100mL×2)萃取,合并后的有机相用无水硫酸钠干燥,减压浓缩后柱层析分离(DCM:甲醇(v/v)=1:0~8:1)得到黄色固体状的20C(0.384g,产率40%)。
LCMS m/z=856.5[M+1]。
第三步:N-[2-[[(2R)-2-羟基-2-(8-羟基-2-氧代-1H-喹啉-5-基)乙基]胺基]乙基]-N-乙基-3-[2-[3-[2-[4-(4-甲基嘧啶基-2-基)-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-基]乙基]苯基]乙氧基]丙酰胺(化合物20)
N-[2-[[(2R)-2-hydroxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]-N-ethyl-3-[2-[3-[2-[4-(4-methylpyrimidin-2-yl)-1-oxa-4,9-diazaspiro[5.5]undecan-9-yl]ethyl]phenyl]ethoxy]propanamide
将20C(0.38g,0.45mmol)溶于THF(10mL)中,加入三乙胺三氢氟酸(0.18g,0.9mmol),室温搅拌12小时。向其中依次滴加饱和碳酸氢钠水溶液(50mL),甲醇(5mL)淬灭反应,用DCM(100mL×2)萃取,合并后的有机相用无水硫酸钠干燥,减压浓缩后柱层析分离(DCM:甲醇(v/v)=1:0~8:1)得到黄色固体状的化合物20(0.12g,产率36%)。
1H NMR(400MHz,CD3OD)δ8.40(d,1H),8.17(d,1H),7.40–7.29(m,5H),7.03(d,1H),6.63–6.65(m,
2H),5.44(d,1H),4.29(dd,2H),3.68–3.81(m,13H),3.20–3.45(m,12H),2.89(t,2H),2.65(t,2H),2.38(s,3H),2.15-2.18(m,2H),1.85(s,2H).1.18-1.41(m,6H).
LCMS m/z=742.5[M+1]。
实施例21:3-[2-[3-[[4-(4-乙基嘧啶-2-基)-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-基]甲基]苯基]乙氧基]-N-[2-[[(2R)-2-羟基-2-(8-羟基-2-氧代-1H-喹啉-5-基)乙基]胺基]乙基]-N-甲基-丙酰胺(化合物21)
3-[2-[3-[[4-(4-ethylpyrimidin-2-yl)-1-oxa-4,9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]-N-[2-[[(2R)-2-hydroxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]-N-methyl-propanamide
第一步:叔丁基3-[2-[3-[[4-(4-乙基嘧啶-2-基)-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-基]甲基]苯基]乙氧基]丙酸酯(21B)
tert-butyl 3-[2-[3-[[4-(4-ethylpyrimidin-2-yl)-1-oxa-4,9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]propanoate
将11B的三氟乙酸盐(0.538g,2.05mmol)置于乙腈(25mL)中,依次加入3-[2-[3-(2-氯甲基)苯基]乙氧基]丙酸叔丁酯(中间体6)(0.613g,2.05mmol),碳酸钾(1.42g,10.3mmol),水(0.5mL),60℃搅拌24小时。待反应冷至室温后,加入水(50mL),用乙酸乙酯(100mL×2)萃取,合并后的有机相用无水硫酸铵干燥,减压浓缩后柱层析分离(DCM/甲醇(v/v)=15:1)得到黄色液体状的21B(0.791g,产率73.5%)。
LCMS m/z=525.3[M+1]。
第二步:3-[2-[3-[[4-(4-乙基嘧啶基-2-基)-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-基]甲基]苯基]乙氧基]丙酸(21C)
3-[2-[3-[[4-(4-ethylpyrimidin-2-yl)-1-oxa-4,9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]propanoicacid
将21B(0.37g,0.71mmol)溶于DCM(15mL)中,滴加三氟乙酸(5mL),室温搅拌4小时。减压浓缩除去反应溶剂,向残余物中加入甲苯(10mL),减压浓缩后得到黄色液体状的21C的三氟乙酸盐(0.33g,产率100%)。
LCMS m/z=469.3[M+1].
第三步:N-(2,2-二甲氧乙基)-N-甲基-3-[2-[3-[[4-(4-乙基嘧啶基-2-基)-1-氧杂-4,9-二氮杂螺[5.5]十一烷
-9-基]甲基]苯基]乙氧基]-丙酰胺(21D)
N-(2,2-dimethoxyethyl)-N-methyl-3-[2-[3-[[4-(4-ethylpyrimidin-2-yl)-1-oxa-4,9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]propanamide
将21C的三氟乙酸盐(0.37g,0.79mmol)溶于DCM(20mL)中,依次加入三乙胺(0.79g,7.8mmol),甲氨基乙醛缩二甲醇(0.11g,0.95mmol),HATU(0.45g,1.2mmol),室温搅拌2小时。滴加饱和碳酸氢钠水溶液(50mL),用DCM(100mL×2)萃取,合并后的有机相用饱和食盐水(50mL)洗涤,无水硫酸钠干燥,减压浓缩后柱层析分离(DCM/甲醇(v/v)=15:1)得到黄色液体状的21D(0.44g,产率98%)。
LCMS m/z=571.3[M+1]。
第四步:N-[2-[[(2R)-2-[叔丁基(二甲基)硅基]氧基-2-(8-羟基-2-氧代-1H-喹啉-5-基)乙基]胺基]乙基-N-甲基-3-[2-[3-[[4-(4-乙基嘧啶基-2-基)-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-基]甲基]苯基]乙氧基]丙酰胺(21E)
N-[2-[[(2R)-2-[tert-butyl(dimethyl)silyl]oxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]-3-[2-[3-[[4-(4-ethylpyrimidin-2-yl)-1-oxa-4,9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]-N-methyl-propanamide
将21D(0.370g,0.65mmol)溶于THF(10mL)中,向其中加入TsOH·H2O(0.62g,3.2mmol),40℃搅拌1小时后,滴加饱和碳酸氢钠水溶液(50mL)淬灭反应,用DCM(100mL×2)萃取,合并后的有机相用无水硫酸钠干燥,减压浓缩后得到的粗产品溶于DCM(10mL)和甲醇(3mL)的混合溶剂中,加入5-[(1R)-2-氨基-1-[叔丁基(二甲基)甲硅烷基]氧基甲基]-8-羟基-1H-喹啉-2-酮(0.22g,0.65mmol),室温搅拌30分钟后加入三乙酰氧基硼氢化钠(0.41g,1.91mmol),室温搅拌3小时。加入饱和碳酸氢钠水溶液(50mL),用DCM(100mL×2)萃取,合并后的有机相用无水硫酸钠干燥,减压浓缩后柱层析分离(DCM:甲醇(v/v)=1:0~8:1)得到黄色固体状的21E(0.22g,产率40%)。
LCMS m/z=842.4[M+1]。
第五步:3-[2-[3-[[4-(4-乙基嘧啶-2-基)-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-基]甲基]苯基]乙氧基]-N-[2-[[(2R)-2-羟基-2-(8-羟基-2-氧代-1H-喹啉-5-基)乙基]胺基]乙基]-N-甲基-丙酰胺(化合物21)
3-[2-[3-[[4-(4-ethylpyrimidin-2-yl)-1-oxa-4,9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]-N-[2-[[(2R)-2-hydroxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]-N-methyl-propanamide
将21E(0.215g,0.255mmol)溶于THF(10mL)中,加入三乙胺三氢氟酸(0.074g,0.38mmol),室温搅拌12小时。依次滴加饱和碳酸氢钠水溶液(50mL),甲醇(5mL)淬灭反应,用DCM(100mL×2)萃取,合并后的有机相用无水硫酸钠干燥,减压浓缩后柱层析分离(DCM:甲醇(v/v)=1:0~8:1)得到黄色固体状的化合物21(0.11g,产率59%)。
1H NMR(400MHz,CD3OD)δ8.42(d,1H),8.16(d,1H),7.48–7.19(m,5H),7.03(d,1H),6.66(d,1H),6.50(d,1H),5.42–5.30(m,1H),4.11(s,2H),3.88–3.58(m,11H),3.47(s,1H),3.27(s,1H),3.20(dq,4H),3.16–2.99(m,8H),2.90(dd,2H),2.72–2.63(m,3H),2.59(dd,2H),2.04(dd,3H),1.87–1.68(m,3H),1.58(s,2H).
LCMS m/z=729.4[M+1]。
实施例22:3-[2-[3-[[4-[1-(环丙甲基)吡唑-3-羰基]-1-氧杂-4,9-杂螺[5.5]十一烷-9-基]甲基]苯基]乙氧基]-N-[2-[[(2R)-2-羟基-2-(5-羟基-3-氧代-4H-1,4-苯并噁嗪-8-基)乙基]胺基]乙基]-N-甲基-丙酰胺(化合物22)
3-[2-[3-[[4-[1-(cyclopropylmethyl)pyrazole-3-carbonyl]-1-oxa-4,9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]-N-[2-[[(2R)-2-hydroxy-2-(5-hydroxy-3-oxo-4H-1,4-benzoxazin-8-yl)ethyl]amino]ethyl]-N-methyl-propanamide
第一步:3-[2-[3-[[4-[1-(环丙甲基)吡唑-3-羰基]-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-基]甲基]苯基]乙氧基]丙酸叔丁酯(22A)
tert-butyl 3-[2-[3-[[4-[1-(cyclopropylmethyl)pyrazole-3-carbonyl]-1-oxa-4,9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]propanoate
将1E的三氟乙酸盐(0.95g,3.1mmol)置于乙腈(25mL)中,依次加入3-[2-[3-(氯甲基)苯基]乙氧基]丙酸叔丁酯(中间体6)(0.93g,3.1mmol),碳酸钾(1.3g,9.4mmol),水(1mL),60℃搅拌3.5小时。待反应冷至室温后,加入水(50mL),用DCM(100mL×2)萃取,合并后的有机相用无水硫酸钠干燥,减压浓缩后柱层析分离(DCM/甲醇(v/v)=15:1)得到黄色液体状的22A(1.65g,产率93%)。
第二步:3-[2-[3-[[4-[1-(环丙甲基)吡唑-3-羰基]-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-基]甲基]苯基]乙氧基]-N-(2,2-二甲氧基乙基)-N-甲基-丙酰胺(22B)
3-[2-[3-[[4-[1-(cyclopropylmethyl)pyrazole-3-carbonyl]-1-oxa-4,9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]-N-(2,2-dimethoxyethyl)-N-methyl-propanamide
将22A(1.65g,2.9mmol)溶于DCM(10mL)中,0℃滴加三氟乙酸(5mL),滴毕,室温反应1小时。反应液直接减压浓缩,加入甲苯(20mL)后再次减压浓缩得到残留物。在0℃和氮气下向残留物中依次加入DCM(15mL)和三乙胺(1.37g,13.5mmol),搅拌均匀后加入N-(2,2-二甲氧基乙基)甲-1-氨(0.39g,3.2mmol)和HATU(1.54g,4.0mmol),室温反应1小时。加入水(30mL),DCM(30mL×2)萃取,合并有机相,饱和食盐水(40mL)洗,无水硫酸钠干燥,过滤,滤液减压浓缩,残留物经硅胶柱层析纯化(DCM/甲醇(v/v)=1:20)得到黄色液体状的22B(1.51g,产率91%)。
第三步:N-[2-[[(2R)-2-[叔丁基(二甲基)硅基]氧基-2-(5-羟基-3-氧杂-4H-1,4-苯并噁嗪-8-基)乙基]氨基]乙基]-3-[2-[3-[[4-[1-(环丙基甲基)吡唑-3-羰基]-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-基]甲基]苯基]乙氧基]-N-甲基-丙酰胺(22C)
N-[2-[[(2R)-2-[tert-butyl(dimethyl)silyl]oxy-2-(5-hydroxy-3-oxo-4H-1,4-benzoxazin-8-yl)ethyl]amino]ethyl]-3-[2-[3-[[4-[1-(cyclopropylmethyl)pyrazole-3-carbonyl]-1-oxa-4,9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]-N-methyl-propanamide
将22B(0.75g,1.2mmol)溶于THF(15mL)中,加入TsOH·H2O(1.2g,6.1mmol),40℃搅拌1小时后,滴加饱和碳酸氢钠水溶液(50mL)淬灭反应,用DCM(100mL×2)萃取,合并后的有机相用无水硫酸钠干燥,减压浓缩后得到的粗产品溶于DCM(10mL)和甲醇(3mL)的混合溶剂中,加入8-[(1R)-2-氨基-1-[叔丁基(二甲基)硅基]氧基-乙基]-5-羟基-4H-1,4-苯并噁嗪-3-酮(0.26g,0.86mmol),室温搅拌30分钟后加入三乙酰氧基硼氢化钠(0.78g,3.7mmol),室温搅拌3小时。加入饱和碳酸氢钠水溶液(50mL),用DCM(100mL×2)萃取,合并后的有机相用无水硫酸钠干燥,减压浓缩后柱层析分离(DCM:甲醇(v/v)=1:0~8:1)得到黄色固体状的22C(0.52g,产率48%)。
第四步:3-[2-[3-[[4-[1-(环丙基甲基)吡唑-3-羰基]-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-基]甲基]苯基]乙氧基]-N-[2-[[(2R)-2-羟基-2-(5-羟基-3-氧代-4H-1,4-苯并噁嗪-8-基)乙基]氨基]乙基]-N-甲基-丙酰胺(化合物22)
3-[2-[3-[[4-[1-(cyclopropylmethyl)pyrazole-3-carbonyl]-1-oxa-4,9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]-N-[2-[[(2R)-2-hydroxy-2-(5-hydroxy-3-oxo-4H-1,4-benzoxazin-8-yl)ethyl]amino]ethyl]-N-methyl-propanamide
将22C(0.52g,0.59mmol)溶于THF(10mL)中,加入三乙胺三氢氟酸(0.98g,6.1mmol),室温搅拌12小时。依次滴加饱和碳酸氢钠水溶液(50mL),甲醇(5mL)淬灭反应,用DCM(100mL×2)萃取,合并后的有机相用无水硫酸钠干燥,减压浓缩后柱层析分离(DCM:甲醇(v/v)=1:0~8:1)得到黄色固体状的化合物22(0.22g,产率49%)。
1H NMR(400MHz,DMSO-d6)δ9.80(br,1H),7.82(d,1H),7.19(m,1H),7.08-7.10(m,3H),6.86(dd,1H),6.55(s,1H),6.49(d,1H),4.99(m,1H),4.84–4.78(m,1H),4.47(t,2H),3.91-4.01(m,4H),3.67–3.51(m,8H),3.22-3.42(m,8H),2.75-2.78(m,4H),2.67(t,1H),2.61(dd,2H),2.37–2.24(m,4H),1.68(d,2H),1.45-1.57(m,2H),0.49-0.57(m,2H),0.32-0.41(m,2H).
LCMS m/z=774.4[M+1].
实施例23:N-[2-[[(2R)-2-羟基-2-(8-羟基-2-氧代-1H-喹啉-5-基)乙基]氨基]乙基]-N-甲基-3-[2-[3-[[4-(4-甲基-2-吡啶基)-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-基]甲基]苯基]乙氧基]丙酰胺(化合物23)
N-[2-[[(2R)-2-hydroxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]-N-methyl-3-[2-[3-[[4-(4-methyl-2-pyridyl)-1-oxa-4,9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]propanamide
第一步:4-(4-甲基-2-吡啶基)-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-碳酸叔丁酯(23B)
tert-butyl 4-(4-methyl-2-pyridyl)-1-oxa-4,9-diazaspiro[5.5]undecane-9-carboxylate
将1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-羧酸叔丁酯(5.12g,20.0mmol)置于1,4-二氧六环(50mL)中,依次加入2-氯-4-甲基-吡啶(23A)(3.83g,30.0mmol),碳酸铯(13.0g,40.0mmol),4,5-双二苯基膦-9,9-二甲基氧杂蒽(1.6g,2.8mmol),醋酸钯(0.3g,1.4mmol),氮气氛回流反应8小时。反应液冷却至室温,加入水(100mL),用乙酸乙酯(100mL×2)萃取,合并后的有机相用饱和食盐水(100mL)洗涤,无水硫酸钠干燥,减压浓缩后柱层析分离(石油醚/乙酸乙酯(v/v)=1:1)得到黄色液体状的23B(3.0g,产率43%)。
第二步:3-[2-[3-[[4-(4-甲基-2-吡啶基)-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-基]甲基]苯基]乙氧基]丙酸叔丁酯(23C)
tert-butyl 3-[2-[3-[[4-(4-methyl-2-pyridyl)-1-oxa-4,9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]propanoate
将23B(1.02g,2.9mmol)加入DCM(20mL),0℃滴加三氟乙酸(3mL),滴毕,室温反应1小时后,将反应液减压浓缩,加入甲苯(20mL)后再次减压浓缩。将浓缩后的残余物置于乙腈(25mL)中,依次加入中间体6(0.98g,3.3mmol),碳酸钾(1.5g,11.0mmol),水(1mL),60℃搅拌3.5小时。待反应冷至室温后,加入水(50mL),用DCM(100mL×2)萃取,合并后的有机相用无水硫酸钠干燥,减压浓缩后柱层析分离(DCM/甲醇(v/v)=15:1)得到黄色液体状的23C(1.42g,产率97%)。
第三步:N-(2,2-二甲氧基乙基)-N-甲基-3-[2-[3-[[4-(4-甲基-2-吡啶基)-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-基]甲基]苯基]乙氧基]丙酰胺(23D)
N-(2,2-dimethoxyethyl)-N-methyl-3-[2-[3-[[4-(4-methyl-2-pyridyl)-1-oxa-4,9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]propanamide
将23C(1.42g,2.79mmol)加入DCM(20mL)中,0℃滴加三氟乙酸(4mL),滴毕,室温反应1小时后,将体系减压浓缩,加入甲苯(20mL)后再次减压浓缩。在0℃和氮气条件下向残留物中依次加入DCM(15mL)和三乙胺(1.39g,13.8mmol),搅拌均匀后加入N-(2,2-二甲氧基乙基)甲-1-氨(0.43g,3.6mmol)和HATU(1.57g,4.1mmol),室温反应1小时。反应结束后加入水(30mL),用DCM(80mL×2)萃取,合并有机相,饱和食盐水(40mL)洗,无水硫酸钠干燥,过滤,滤液减压浓缩,残留物经硅胶柱层析纯化(DCM/甲醇(v/v)=1:20)得到黄色液体状的23D(1.11g,产率73%)。
LCMS m/z=555.3[M+1]。
第四步:N-[2-[[(2R)-2-[叔丁基(二甲基)硅基]氧基-2-(8-羟基-2-氧代-1H-喹啉-5-基)乙基]氨基]乙基]-N-甲基-3-[2-[3-[[4-(4-甲基-2-吡啶基)-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-基]甲基]苯基]乙氧基]丙酰胺(23E)
N-[2-[[(2R)-2-[tert-butyl(dimethyl)silyl]oxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]-N-methyl-3-[2-[3-[[4-(4-methyl-2-pyridyl)-1-oxa-4,9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]propanamide
将23D(0.55g,1.0mmol)溶于THF(15mL)中,向其中加入TsOH·H2O(0.94g,5.0mmol),40℃搅拌1小时后,滴加饱和碳酸氢钠水溶液(50mL)淬灭反应,用DCM(100mL×2)萃取,合并后的有机相用无水硫酸钠干燥,减压浓缩后得到的粗产品溶于DCM(10mL)和甲醇(3mL)的混合溶剂中,加入5-[(1R)-2-氨基-1-[叔丁基(二甲基)甲硅烷基]氧基甲基]-8-羟基-1H-喹啉-2-酮(0.23g,0.69mmol),室温搅拌30分钟后加入三乙酰氧基硼氢化钠(0.63g,3.0mmol),室温搅拌3小时。加入饱和碳酸氢钠水溶液(50mL),用DCM(100mL×2)萃取,合并后的有机相用无水硫酸钠干燥,减压浓缩后柱层析分离(DCM:甲醇(v/v)=1:0~8:1)得到黄色固体状的23E(0.23g,产率28%)。
第五步:N-[2-[[(2R)-2-羟基-2-(8-羟基-2-氧代-1H-喹啉-5-基)乙基]氨基]乙基]-N-甲基-3-[2-[3-[[4-(4-甲基-2-吡啶基)-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-基]甲基]苯基]乙氧基]丙酰胺(化合物23)
N-[2-[[(2R)-2-hydroxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]-N-methyl-3-[2-[3-[[4-(4-methyl-2-pyridyl)-1-oxa-4,9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]propanamide
将23E(0.23g,0.28mmol)溶于THF(10mL)中,加入三乙胺三氢氟酸(0.8g,5.0mmol),室温搅拌12小时。依次滴加饱和碳酸氢钠水溶液(50mL),甲醇(5mL)淬灭反应,用DCM(100mL×2)萃取,合并后的有机相用无水硫酸钠干燥,减压浓缩后柱层析分离(DCM:甲醇(v/v)=1:0~8:1)得到黄色固体状的化合物23(0.14g,产率70.8%)。
1H NMR(400MHz,DMSO-d6)δ10.29(s,1H),8.17(d,1H),7.94(d,1H),7.23–7.16(m,1H),7.14(s,1H),7.05-7.11(m,3H),6.91(d,1H),6.65(s,1H),6.51–6.44(m,2H),5.01(t,1H),3.69–3.65(m,2H),3.62–3.54(m,4H),3.44–3.23(m,12H),2.75-2.77(m,3H),2.73–2.60(m,4H),2.41-2.44(m,2H),2.28(t,2H),2.20(s,3H),1.78–1.70(m,2H),1.54(dt,2H).
LCMS m/z=357.3[M/2+1].
实施例24:3-[2-[4-[[4-[1-(环丙甲基)吡唑-3-羰基]-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-基]甲基]苯基]乙氧基]-N-[2-[[(2R)-2-羟基-2-(8-羟基-2-氧代-1H-喹啉-5-基)乙基]氨基]乙基]-N-甲基-丙酰胺(化合物24)
3-[2-[4-[[4-[1-(cyclopropylmethyl)pyrazole-3-carbonyl]-1-oxa-4,9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]-N-[2-[[(2R)-2-hydroxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]-N-methyl-propanamide
第一步:3-[2-[4-[[4-[1-(环丙甲基)吡唑-3-羰基]-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-基]甲基]苯基]乙氧基]丙酸叔丁酯(24A)
tert-butyl 3-[2-[4-[[4-[1-(cyclopropylmethyl)pyrazole-3-carbonyl]-1-oxa-4,9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]propanoate
将1E的三氟乙酸盐(0.38g,1.2mmol)置于乙腈(25mL)中,依次加入3-[2-[4-(溴甲基)苯基]乙氧基]丙酸叔丁酯(中间体2)(0.45g,1.3mmol),碳酸钾(0.35g,2.5mmol),水(1mL),60℃搅拌3.5小时。待反应冷至室温后,加入水(50mL),用DCM(100mL×2)萃取,合并后的有机相用无水硫酸钠干燥,减压浓缩后柱层析分离(DCM/甲醇(v/v)=15:1)得到黄色液体状的24A(0.48g,产率68%)。
第二步:3-[2-[4-[[4-[1-(环丙甲基)吡唑-3-羰基]-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-基]甲基]苯基]乙氧基]-N-(2,2-二甲氧基乙基)-N-甲基-丙酰胺(24B)
3-[2-[4-[[4-[1-(cyclopropylmethyl)pyrazole-3-carbonyl]-1-oxa-4,9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]-N-(2,2-dimethoxyethyl)-N-methyl-propanamide
将24A(0.48g,0.85mmol)加入DCM(10mL),0℃滴加三氟乙酸(2mL),滴毕,室温反应1小时后,将体系减压浓缩,加入甲苯(20mL)后再次减压浓缩得到残留物。在0℃和氮气条件下向残留物中依次加入DCM(15mL)和三乙胺(0.41g,4.0mmol),搅拌均匀后加入N-(2,2-二甲氧基乙基)甲-1-氨(0.14g,1.2mmol)和HATU(0.46g,1.2mmol),升至室温反应1小时。向反应液中加入水(30mL),用DCM(30mL×2)萃取,合并有机相,饱和食盐水(40mL)洗涤,无水硫酸钠干燥,过滤,滤液减压浓缩,残留物经硅胶柱层析纯化(DCM/甲醇(v/v)=1:20)得到黄色液体状的24B(0.42g,产率85%)。
第三步:N-[2-[[(2R)-2-[叔丁基(二甲基)硅基]氧基-2-(8-羟基-2-氧代-1H-喹啉-5-基)乙基]氨基]乙基]-3-[2-[4-[[4-[1-(环丙甲基)吡唑-3-羰基]-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-基]甲基]苯基]乙氧基]-N-甲基-丙酰胺(24C)
N-[2-[[(2R)-2-[tert-butyl(dimethyl)silyl]oxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]-3-[2-[4-[[4-[1-(cyclopropylmethyl)pyrazole-3-carbonyl]-1-oxa-4,9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]-N-methyl-propanamide
将24B(0.42g,0.69mmol)溶于THF(15mL)中,向其中加入TsOH·H2O(0.98g,3.6mmol),40℃搅拌1小时后,滴加饱和碳酸氢钠水溶液(50mL)淬灭反应,用DCM(100mL×2)萃取,合并后的有机相用无水硫酸钠干燥,减压浓缩后得到的粗产品溶于DCM(10mL)和甲醇(3mL)的混合溶剂中,加入5-[(1R)-2-氨基-1-[叔丁基(二甲基)甲硅烷基]氧基甲基]-8-羟基-1H-喹啉-2-酮(0.18g,0.55mmol),室温搅拌30分钟后加入三乙酰氧基硼氢化钠(0.44g,2.1mmol),室温搅拌3小时。加入饱和碳酸氢钠水溶液(50mL),用DCM(100mL×2)萃取,合并后的有机相用无水硫酸钠干燥,减压浓缩后柱层析分离(DCM:甲醇(v/v)=1:0~8:1)得到黄色固体状的24C(0.23g,产率37%)。
LCMS m/z=442.9[M/2+1]。
第四步:3-[2-[4-[[4-[1-(环丙甲基)吡唑-3-羰基]-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-基]甲基]苯基]乙氧基]-N-[2-[[(2R)-2-羟基-2-(8-羟基-2-氧代-1H-喹啉-5-基)乙基]氨基]乙基]-N-甲基-丙酰胺(化合物24)
3-[2-[4-[[4-[1-(cyclopropylmethyl)pyrazole-3-carbonyl]-1-oxa-4,9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]-N-[2-[[(2R)-2-hydroxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]-N-methyl-propanamide
将24C(0.23g,0.30mmol)溶于THF(10mL)中,加入三乙胺三氢氟酸(0.99g,6.1mmol),室温搅拌12小时。向其中依次滴加饱和碳酸氢钠水溶液(50mL),甲醇(5mL)淬灭反应,用DCM(100mL×2)萃取,合并后的有机相用无水硫酸钠干燥,减压浓缩后柱层析分离(DCM:甲醇(v/v)=1:0~8:1)得到黄色固体状的化合物24(0.11g,产率49%)。
1H NMR(400MHz,DMSO-d6)δ10.22(br,1H),8.17(d,1H),7.82(d,1H),7.14(m,4H),7.06(d,1H),6.91(d,1H),6.55(br,1H),6.48(dd,1H),5.00(dd,1H),4.01(m,2H),3.90-3.94(m,2H),3.66–3.52(m,8H),3.42–3.23(m,9H),2.78–2.71(m,4H),2.62-2.69(m,3H),2.26-2.33(m,4H),1.64-1.68(m,2H),1.48(m,2H),0.50-0.57(m,2H),0.40-0.34(m,2H).
LCMS m/z=770.3[M+1]。
实施例25:3-[2-[3-[[4-[1-(环丙基甲基)吡唑-3-羰基]-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-基]甲基]苯基]乙氧基]-N-乙基-N-[2-[[(2R)-2-羟基-2-(8-羟基-2-氧代-1H-喹啉-5-基)乙基]氨基]乙基]丙酰胺(化合物25)
3-[2-[3-[[4-[1-(cyclopropylmethyl)pyrazole-3-carbonyl]-1-oxa-4,9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]-N-ethyl-N-[2-[[(2R)-2-hydroxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]propanamide
第一步:3-[2-[3-[[4-[1-(环丙甲基)吡唑-3-羰基]-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-基]甲基]苯基]乙氧基]-N-(2,2-二甲氧基乙基)-N-乙基-丙酰胺(25A)
3-[2-[3-[[4-[1-(cyclopropylmethyl)pyrazole-3-carbonyl]-1-oxa-4,9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]-N-(2,2-dimethoxyethyl)-N-ethyl-propanamide
将24A(0.39g,0.69mmol)加入DCM(10mL),0℃滴加三氟乙酸(5mL),滴毕,室温反应1小时后,将体系减压浓缩,加入甲苯(20mL)后再次减压浓缩得到残留物。在0℃和氮气条件下向残留物中依次加入DCM(15mL)和三乙胺(0.45g,4.4mmol),搅拌均匀后加入N-(2,2-二甲氧基乙基)乙-1-氨(0.35g,2.6mmol)和HATU(0.60g,1.6mmol),升至室温反应1小时。反应结束后加入水(30mL),用DCM(30mL×2)萃取,合并有机相,饱和食盐水(40mL)洗,无水硫酸钠干燥,过滤,滤液减压浓缩,残留物经硅胶柱层析纯化(DCM/甲醇(v/v)=1:20)得到黄色液体状的25A(0.37g,产率88%)。
第二步:N-[2-[[(2R)-2-[叔丁基(二甲基)硅基]氧基-2-(8-羟基-2-氧代-1H-喹啉-5-基)乙基]氨基]乙基]-3-[2-[3-[[4-[1-(环丙基甲基)吡唑-3-羰基]-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-基]甲基]苯基]乙氧基]-N-乙基-丙酰胺(25B)
N-[2-[[(2R)-2-[tert-butyl(dimethyl)silyl]oxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]-3-[2-[3-[[4-[1-(cyclopropylmethyl)pyrazole-3-carbonyl]-1-oxa-4,9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]-N-ethyl-propanamide
将25A(0.37g,0.58mmol)溶于THF(15mL)中,加入TsOH·H2O(0.56g,2.9mmol),40℃搅拌1小时后,滴加饱和碳酸氢钠水溶液(50mL)淬灭反应,用DCM(100mL×2)萃取,合并后的有机相用无水硫酸钠干燥,减压浓缩后得到的粗产品溶于DCM(10mL)和甲醇(3mL)的混合溶剂中,加入5-[(1R)-2-氨基-1-[叔丁基(二甲基)甲硅烷基]氧基甲基]-8-羟基-1H-喹啉-2-酮(0.16g,0.47mmol),室温搅拌30分钟后加入三乙酰氧基硼氢化钠(0.37g,1.75mmol),室温搅拌3小时。加入饱和碳酸氢钠水溶液(50mL),用DCM(100mL×2)萃取,合并后的有机相用无水硫酸钠干燥,减压浓缩后柱层析分离(DCM:甲醇(v/v)=1:0~8:1)得到黄色固体状的25B(0.27g,产率51%)。
LCMS m/z=449.9[M/2+1].
第三步:3-[2-[3-[[4-[1-(环丙基甲基)吡唑-3-羰基]-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-基]甲基]苯基]乙氧基]-N-乙基-N-[2-[[(2R)-2-羟基-2-(8-羟基-2-氧代-1H-喹啉-5-基)乙基]氨基]乙基]丙酰胺(化合物25)
3-[2-[3-[[4-[1-(cyclopropylmethyl)pyrazole-3-carbonyl]-1-oxa-4,9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]-N-ethyl-N-[2-[[(2R)-2-hydroxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]propanamide
将25B(0.27g,0.30mmol)溶于THF(10mL)中,加入三乙胺三氢氟酸(0.80g,5.0mmol),室温搅拌12小时。依次滴加饱和碳酸氢钠水溶液(50mL),甲醇(5mL)淬灭反应,用DCM(100mL×2)萃取,合并后的有机相用无水硫酸钠干燥,减压浓缩后柱层析分离(DCM:甲醇(v/v)=1:0~8:1)得到黄色固体状的化合物25(0.10g,产率44%)。
1H NMR(400MHz,DMSO-d6)δ10.22(br,1H),8.17(d,1H),7.82(d,1H),7.04-7.18(m,5H),6.91(d,1H),6.55(s,1H),6.50–6.45(m,1H),4.99-5.00(m,1H),4.01(s,2H),3.91-3.93(m,2H),3.66–3.50(m,8H),3.43–3.18(m,11H),2.87–2.56(m,7H),2.27-2.33(m,4H),1.67(d,2H),1.44-1.55(m,2H),0.49-0.57(m,2H),0.33-0.40(m,2H).
LCMS m/z=784.3[M+1].
实施例26:3-[2-[3-[[4-[1-(环丙基甲基)吡唑-3-羰基]-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-基]甲基]苯基]
乙氧基]-N-[2-[[(2R)-2-羟基-2-(8-羟基-2-氧代-1H-喹啉-5-基)乙基]氨基]乙基]-N-丙基-丙酰胺(化合物26)
3-[2-[3-[[4-[1-(cyclopropylmethyl)pyrazole-3-carbonyl]-1-oxa-4,9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]-N-[2-[[(2R)-2-hydroxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]-N-propyl-propanamide
第一步:3-[2-[3-[[4-[1-(环丙甲基)吡唑-3-羰基]-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-基]甲基]苯基]乙氧基]-N-(2,2-二甲氧基乙基)-N-丙基-丙酰胺(26A)
3-[2-[3-[[4-[1-(cyclopropylmethyl)pyrazole-3-carbonyl]-1-oxa-4,9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]-N-(2,2-dimethoxyethyl)-N-propyl-propanamide
将24A(0.39g,0.63mmol)加入DCM(10mL)中,0℃滴加三氟乙酸(2mL),滴毕,室温反应1小时后,将体系减压浓缩,加入甲苯(20mL)后再次减压浓缩得到残留物。在0℃和氮气条件下向残留物中依次加入DCM(15mL)和三乙胺(0.45g,4.4mmol),搅拌均匀后加入N-(2,2-二甲氧基乙基)丙-1-氨(0.36g,2.4mmol)和HATU(0.60g,1.6mmol),升至室温反应1小时。反应结束后加入水(30mL),用DCM(50mL×2)萃取,合并有机相,饱和食盐水(40mL)洗涤,无水硫酸钠干燥,过滤,滤液减压浓缩,残留物经硅胶柱层析纯化(DCM/甲醇(v/v)=1:20)得到黄色液体状的26A(0.36g,产率84%)。
第二步:N-[2-[[(2R)-2-[叔丁基(二甲基)硅基]氧基-2-(8-羟基-2-氧代-1H-喹啉-5-基)乙基]氨基]乙基]-3-[2-[3-[[4-[1-(环丙基甲基)吡唑-3-羰基]-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-基]甲基]苯基]乙氧基]-N-丙基-丙酰胺(26B)
N-[2-[[(2R)-2-[tert-butyl(dimethyl)silyl]oxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]-3-[2-[3-[[4-[1-(cyclopropylmethyl)pyrazole-3-carbonyl]-1-oxa-4,9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]-N-propyl-propanamide
将26A(0.36g,0.56mmol)溶于THF(15mL)中,加入TsOH·H2O(0.53g,2.8mmol),40℃搅拌1小时后,滴加饱和碳酸氢钠水溶液(50mL)淬灭反应,用DCM(100mL×2)萃取,合并后的有机相用无水硫酸钠干燥,减压浓缩后得到的粗产品溶于DCM(10mL)和甲醇(3mL)的混合溶剂中,加入5-[(1R)-2-氨基-1-[叔丁基(二甲基)甲硅烷基]氧基甲基]-8-羟基-1H-喹啉-2-酮(0.15g,0.45mmol),室温搅拌30分钟后加
入三乙酰氧基硼氢化钠(0.36g,1.7mmol),室温搅拌3小时。加入饱和碳酸氢钠水溶液(50mL),用DCM(100mL×2)萃取,合并后的有机相用无水硫酸钠干燥,减压浓缩后柱层析分离(DCM:甲醇(v/v)=1:0~8:1)得到黄色固体状的26B(0.20g,产率39%)。
LCMS m/z=456.7[M/2+1]。
第三步:3-[2-[3-[[4-[1-(环丙基甲基)吡唑-3-羰基]-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-基]甲基]苯基]乙氧基]-N-[2-[[(2R)-2-羟基-2-(8-羟基-2-氧代-1H-喹啉-5-基)乙基]氨基]乙基]-N-丙基-丙酰胺(化合物26)
3-[2-[3-[[4-[1-(cyclopropylmethyl)pyrazole-3-carbonyl]-1-oxa-4,9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]-N-[2-[[(2R)-2-hydroxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]-N-propyl-propanamide
将26B(0.20g,0.22mmol)溶于THF(10mL)中,加入三乙胺三氢氟酸(0.98g,6.1mmol),室温搅拌12小时。依次滴加饱和碳酸氢钠水溶液(50mL),甲醇(5mL)淬灭反应,用DCM(100mL×2)萃取,合并后的有机相用无水硫酸钠干燥,减压浓缩后柱层析分离(DCM:甲醇(v/v)=1:0~8:1)得到黄色固体状的化合物26(0.13g,产率72%)。
1H NMR(400MHz,DMSO-d6)δ10.24(s,1H),8.18(d,1H),7.82(d,1H),7.04-7.09(m,5H),6.91(d,1H),6.55(s,1H),6.50–6.45(m,1H),5.04–4.94(m,1H),4.01(m,2H),3.91-3.94(m,2H),3.50-3.61(m,8H),3.46–3.22(m,14H),2.78–2.59(m,6H),2.28-2.33(m,4H),1.67(d,2H),1.45-1.49(m,2H),0.50-0.58(m,2H),0.33-0.41(m,2H).
LCMS m/z=798.5[M+1].
实施例27:3-[2-[3-[2-[4-[1-(环丙基甲基)吡唑-3-羰基]-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-基]乙基]苯基]乙氧基]-N-乙基-N-[2-[[(2R)-2-羟基-2-(8-羟基-2-氧代-1H-喹啉-5-基)乙基]氨基]乙基]丙酰胺(化合物27)
3-[2-[3-[2-[4-[1-(cyclopropylmethyl)pyrazole-3-carbonyl]-1-oxa-4,9-diazaspiro[5.5]undecan-9-yl]ethyl]phenyl]ethoxy]-N-ethyl-N-[2-[[(2R)-2-hydroxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]propanamide
第一步:3-[2-[3-[2-[4-[1-(环丙甲基)吡唑-3-羰基]-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-基]乙基]苯基]乙氧基]-N-(2,2-二甲氧基乙基)-N-乙基-丙酰胺(27A)
3-[2-[3-[2-[4-[1-(cyclopropylmethyl)pyrazole-3-carbonyl]-1-oxa-4,9-diazaspiro[5.5]undecan-9-yl]ethyl]phenyl]ethoxy]-N-(2,2-dimethoxyethyl)-N-ethyl-propanamide
将1F(0.71g,1.2mmol)加入DCM(10mL)中,0℃滴加三氟乙酸(2mL),滴毕,室温反应1小时后,将体系减压浓缩,加入甲苯(20mL)后再次减压浓缩得到残留物。在0℃和氮气条件下向残留物中依次加入DCM(15mL)和三乙胺(0.59g,5.8mmol),搅拌均匀后加入N-(2,2-二甲氧基乙基)乙-1-氨(0.17g,1.3mmol)和HATU(0.66g,1.7mmol),升至室温反应1小时。反应结束后加入水(30mL),用DCM(30mL×2)萃取,合并有机相,饱和食盐水(40mL)洗,无水硫酸钠干燥,过滤,滤液减压浓缩,残留物经硅胶柱层析纯化(DCM/甲醇(v/v)=1:20)得到黄色液体状的27A(0.70g,产率:94%)。
LCMS m/z=640.4[M+1]。
第二步:N-[2-[[(2R)-2-[叔丁基(二甲基)硅基]氧-2-(8-羟基-2-氧代-1H-喹啉-5-基)乙基]氨基]乙基]-3-[2-[3-[2-[4-[1-(环丙基甲基)吡唑-3-羰基]-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-基]乙基]苯基]乙氧基]-N-乙基-丙酰胺(27B)
N-[2-[[(2R)-2-[tert-butyl(dimethyl)silyl]oxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]-3-[2-[3-[2-[4-[1-(cyclopropylmethyl)pyrazole-3-carbonyl]-1-oxa-4,9-diazaspiro[5.5]undecan-9-yl]ethyl]phenyl]ethoxy]-N-ethyl-propanamide
将27A(0.37g,0.58mmol)溶于THF(15mL)中,向其中加入TsOH·H2O(0.61g,3.2mmol),40℃搅拌1小时后,滴加饱和碳酸氢钠水溶液(50mL)淬灭反应,用DCM(100mL×2)萃取,合并后的有机相用无水硫酸钠干燥,减压浓缩后得到的粗产品溶于DCM(10mL)和甲醇(3mL)的混合溶剂中,加入5-[(1R)-2-氨基-1-[叔丁基(二甲基)甲硅烷基]氧基甲基]-8-羟基-1H-喹啉-2-酮(0.2g,0.6mmol),室温搅拌30分钟后加入三乙酰氧基硼氢化钠(0.37g,1.7mmol),室温搅拌3小时。加入饱和碳酸氢钠水溶液(50mL),用DCM(100mL×2)萃取,合并后的有机相用无水硫酸钠干燥,减压浓缩后柱层析分离(DCM:甲醇(v/v)=1:0~8:1)得到黄色固体状的27B(0.20g,产率38%)。
LCMS m/z=456.9[M/2+1]。
第三步:3-[2-[3-[2-[4-[1-(环丙基甲基)吡唑-3-羰基]-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-基]乙基]苯基]乙氧基]-N-乙基-N-[2-[[(2R)-2-羟基-2-(8-羟基-2-氧代-1H-喹啉-5-基)乙基]氨基]乙基]丙酰胺(化合物27)
3-[2-[3-[2-[4-[1-(cyclopropylmethyl)pyrazole-3-carbonyl]-1-oxa-4,9-diazaspiro[5.5]undecan-9-yl]ethyl]phenyl]ethoxy]-N-ethyl-N-[2-[[(2R)-2-hydroxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]propanamide
将27B(0.20g,0.22mmol)溶于THF(10mL)中,加入三乙胺三氢氟酸(0.99g,6.1mmol),室温搅拌12小时。向其中依次滴加饱和碳酸氢钠水溶液(50mL),甲醇(5mL)淬灭反应,用DCM(100mL×2)萃取,合并后的有机相用无水硫酸钠干燥,减压浓缩后柱层析分离(DCM:甲醇(v/v)=1:0~8:1)得到黄色固体状的化合物27(0.125g,产率71%)。
1H NMR(400MHz,DMSO-d6)δ10.27(br,1H),8.18(dd,1H),7.82(s,1H),7.17–7.11(m,1H),6.99-7.08(m,4H),6.92(dd,1H),6.57(s,1H),6.49(dd,1H),4.99-5.09(m,1H),4.01(d,2H),3.94(s,2H),3.67–3.50(m,9H),3.18-3.42(m,9H),2.78–2.59(m,11H),2.31-2.43(m,4H),1.63-1.71(m,2H),1.48-1.57(m,2H),0.49-0.55(m,2H),0.38–0.34(m,2H).
LCMS m/z=798.5[M+1]。
实施例28:3-[2-[3-[[4-[1-(环丙基甲基)吡唑-3-羰基]-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-基]甲基]苯基]乙氧基]-N-[2-[2-(5-羟基-3-氧-4H-1,4-苯并噁嗪-8-基)乙基氨基]乙基]-N-甲基-丙酰胺二三氟乙酸盐(化合物28)
3-[2-[3-[[4-[1-(cyclopropylmethyl)pyrazole-3-carbonyl]-1-oxa-4,9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]-N-[2-[2-(5-hydroxy-3-oxo-4H-1,4-benzoxazin-8-yl)ethylamino]ethyl]-N-methyl-propanamide ditrifluoroacetic acid
将24B(0.37g,0.60mmol)溶于N-甲基吡咯烷酮(15mL)中,依次加入8-(2-胺乙基)-5-羟基-4H-1,4-苯并噁嗪-3-酮的乙酸盐(0.15g,0.72mmol),乙酸(0.037g,0.61mmol)。室温搅拌30分钟后,加入三乙酰氧基硼氢化钠(0.37g,1.7mmol),5小时后终止反应。减压浓缩除去大部分反应溶剂,向残余物中缓慢滴加饱和碳酸氢钠水溶液(50mL),用DCM萃取(100mL×2),合并后的有机相用饱和食盐水洗(50mL),无水硫酸钠干燥后,减压浓缩柱层析分离[(DCM/甲醇(v/v)=15:1)]后得到粗品,粗品用液相制备柱分离(液相制备条件:C18反相制备柱,流动相为含0.1%三氟乙酸的去离子水(A),含0.1%三氟乙酸的乙腈(B),梯度洗脱B:(A+B)=5%~20%,洗脱时间15min,流速12mL/min,柱温:30℃)得到白色固体状的化合物28(0.034g,产率7%)。
1H NMR(400MHz,DMSO-d6)δ9.78(br,1H),7.82(d,1H),7.22-7.16(m,1H),7.14-7.05(m,3H),6.60(d,1H),6.55(s,1H),6.45–6.38(m,1H),4.46(d,2H),4.04-3.98(m,2H),3.96-3.87(m,2H),3.65–3.50(m,9H),3.43–3.26(m,11H),2.66–2.53(m,6H),2.38–2.25(m,4H),1.70-1.63(m,2H),1.56-1.46(m,2H),0.57-0.50(m,2H),0.40-0.34(m,2H).
LCMS m/z=758.4[M+1]。
实施例29:N-环戊基-3-[2-[3-[[4-[1-(环丙甲基)吡唑-3-羰基]-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-基]甲基]苯基]乙氧基]-N-[2-[2-(5-羟基-3-氧-4H-1,4-苯并噁嗪-8-基)乙基氨基]乙基]丙酰胺二三氟乙酸盐(化合物29)
N-cyclopentyl-3-[2-[3-[[4-[1-(cyclopropylmethyl)pyrazole-3-carbonyl]-1-oxa-4,9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]-N-[2-[2-(5-hydroxy-3-oxo-4H-1,4-benzoxazin-8-yl)ethylamino]ethyl]propanamide ditrifluoroacetic acid
第一步:N-环戊基-3-[2-[3-[[4-[1-(环丙甲基)吡唑-3-羰基]-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-基]甲基]
苯基]乙氧基]-N-(2,2-二甲氧乙基)丙酰胺(29A)
N-cyclopentyl-3-[2-[3-[[4-[1-(cyclopropylmethyl)pyrazole-3-carbonyl]-1-oxa-4,9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]-N-(2,2-dimethoxyethyl)propanamide
将24A(0.39g,0.67mmol)加入DCM(10mL)中,0℃滴加三氟乙酸(2mL),滴毕,室温反应1小时后,将体系减压浓缩,加入甲苯(20mL)后再次减压浓缩得到残留物。在0℃和氮气条件下向残留物中依次加入DCM(15mL)和三乙胺(0.45g,4.4mmol),搅拌均匀后加入N-(2,2-二甲氧乙基)环戊胺(0.40g,2.3mmol)和HATU(0.60g,1.6mmol),升至室温反应1小时。反应结束后加入水(30mL),用DCM(30mL×2)萃取,合并有机相,饱和食盐水(40mL)洗,无水硫酸钠干燥,过滤,滤液减压浓缩,残留物经硅胶柱层析纯化(DCM/甲醇(v/v)=1:20)得到黄色液体状的29A(0.40g,产率91%)。
第二步:N-环戊基-3-[2-[3-[[4-[1-(环丙甲基)吡唑-3-羰基]-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-基]甲基]苯基]乙氧基]-N-[2-[2-(5-羟基-3-氧-4H-1,4-苯并噁嗪-8-基)乙基氨基]乙基]丙酰胺二三氟乙酸盐(化合物29)
N-cyclopentyl-3-[2-[3-[[4-[1-(cyclopropylmethyl)pyrazole-3-carbonyl]-1-oxa-4,9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]-N-[2-[2-(5-hydroxy-3-oxo-4H-1,4-benzoxazin-8-yl)ethylamino]ethyl]propanamide ditrifluoroacetic acid
将29A(0.40g,0.61mmol)溶于N-甲基吡咯烷酮(15mL)中,依次加入8-(2-胺乙基)-5-羟基-4H-1,4-苯并噁嗪-3-酮的乙酸盐(0.15g,0.72mmol),乙酸(0.04g,0.7mmol)。室温搅拌30分钟后,加入三乙酰氧基硼氢化钠(0.40g,1.9mmol),5小时后终止反应。减压浓缩除去大部分反应溶剂,向残余物中缓慢滴加饱和碳酸氢钠水溶液(50mL),用DCM萃取(100mL×2),合并后的有机相用饱和食盐水(50mL)洗涤,无水硫酸钠干燥后,减压浓缩后的残留物经硅胶柱层析纯化(DCM/甲醇(v/)=1:10)后得到粗品,粗品用柱色谱制备(液相制备条件:C18反相制备柱,流动相为含0.1%三氟乙酸的去离子水(A),含0.1%三氟乙酸的乙腈(B),梯度洗脱B:(A+B)=5%~20%,洗脱时间15min,流速12mL/min,柱温:30℃)得到白色固体状的化合物29(0.029g,产率6%)。
1H NMR(400MHz,CD3OD)δ7.71(d,1H),7.40-7.31(m,4H),6.73(d,1H),6.63(d,1H),6.49(d,1H),4.62(s,2H),4.30-4.26(m,2H),4.09–3.98(m,4H),3.82-3.73(m,3H),3.70(dd,4H),3.65–3.62(m,1H),3.50(t,2H),3.39-3.31(m,2H),3.25-3.16(m,4H),3.12(t,2H),2.93-2.87(m,4H),2.68(t,2H),2.24-2.16(m,2H),1.91-1.83(m,2H),1.81–1.70(m,4H),1.64-1.57(m,2H),1.53–1.45(m,2H),0.99–0.82(m,2H),0.59(d,2H),0.38(s,2H).
LCMS m/z=812.5[M+1]。
实施例30:N-[2-[[(2R)-2-羟基-2-(8-羟基-2-氧代-1H-喹啉-5-基)乙基]胺基]乙基]-3-[2-[3-[[4-(4-甲氧基嘧啶基-2-基)-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-基]甲基]苯基]乙氧基]-N-甲基-丙酰胺(化合物30)
N-[2-[[(2R)-2-hydroxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]-3-[2-[3-[[4-(4-methoxypyrimidin-2-yl)-1-oxa-4,9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]-N-methyl-propanamide
第一步:4-(4-甲基嘧啶基-2-基)-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-羧酸叔丁酯(30B)
tert-butyl 4-(4-methoxypyrimidin-2-yl)-1-oxa-4,9-diazaspiro[5.5]undecane-9-carboxylate
将1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-羧酸叔丁酯(0.769g,3.00mmol)溶于DMF(20mL)中,依次加入2-溴3-甲氧基嘧啶(30A)(0.567g,3.00mmol)、碳酸钾(0.829g,6.00mmol),90℃搅拌4小时。加入乙酸乙酯(200mL),水(100mL),萃取,有机相和水相分离,水相用乙酸乙酯(100mL×1)萃取,合并后的有机相用饱和食盐水(100mL)洗涤,无水硫酸钠干燥,减压浓缩后柱层析分离(石油醚/乙酸乙酯(v/v)=1:1)得到黄色液体状的30B(0.961g,产率87.9%)。
LCMS m/z=365.3[M+1]。
第二步:4-(4-甲氧基嘧啶基-2-基)-1-氧杂-4,9-二氮杂螺[5.5]十一烷(30C)
4-(4-methoxypyrimidin-2-yl)-1-oxa-4,9-diazaspiro[5.5]undecane
将30B(1.367g,1.77mmol)溶于DCM(10mL)中,加入三氟乙酸(4mL),室温搅拌2小时。减压浓缩除去反应溶剂,向残余物中加入甲苯(5mL),减压浓缩后得到黄色液体状30C的三氟乙酸盐(0.99g,产率100%)。
LCMS m/z=265.3[M+1]。
第三步:3-[2-[3-[[4-(4-甲氧基嘧啶-2-基)-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-基]甲基]苯基]乙氧基]丙酸叔丁酯(30D)
tert-butyl 3-[2-[3-[[4-(4-methoxypyrimidin-2-yl)-1-oxa-4,9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]propanoate
将30C的三氟乙酸盐(0.99g,3.75mmol)置于乙腈(25mL)中,依次加入中间体2(1.12g,3.75mmol),三乙胺(1.221g,12.07mmol),60℃搅拌24小时。待反应冷至室温后,加入水(50mL),用乙酸乙酯(100mL×2)萃取,合并后的有机相用无水硫酸钠干燥,减压浓缩后柱层析分离(DCM/甲醇(v/v)=15:1)得到
黄色液体状的30D(1.967g,产率100%)。
LCMS m/z=527.3[M+1]。
第四步:3-[2-[3-[[4-(4-甲氧基嘧啶基-2-基)-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-基]甲基]苯基]乙氧基]丙酸(30E)
3-[2-[3-[[4-(4-methoxypyrimidin-2-yl)-1-oxa-4,9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]propanoic acid
将30D(1.90g,3.61mmol)溶于DCM(15mL)中,滴加三氟乙酸(5mL),室温搅拌4小时。减压浓缩除去反应溶剂,向残余物中加入甲苯(10mL),减压浓缩后得到黄色液体状的30E的三氟乙酸盐(1.70g,产率100%)。
LCMS m/z=471.4[M+1]。
第五步:N-(2,2-二甲氧乙基)-3-[2-[3-[[4-(4-甲氧基嘧啶基-2-基)-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-基]甲基]苯基]乙氧基]-N-甲基-丙酰胺(30F)
N-(2,2-dimethoxyethyl)-3-[2-[3-[2-[4-(4-methoxypyrimidin-2-yl)-1-oxa-4,9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]-N-methyl-propanamide
将30E的三氟乙酸盐(3.61g,1.70mmol)溶于DCM(20mL)中,依次加入三乙胺(0.73g,7.22mmol),甲氨基乙醛缩二甲醇(0.52g,4.33mmol),HATU(2.06g,5.42mmol),室温搅拌2小时。滴加饱和碳酸氢钠水溶液(50mL),用DCM(100mL×2)萃取,合并后的有机相用饱和食盐水(50mL)洗涤,无水硫酸钠干燥,减压浓缩后柱层析分离(DCM/甲醇(v/v)=15:1)得到黄色液体状的30F(2.06g,产率100%)。
LCMS m/z=572.3[M+1]。
第六步:N-[2-[[(2R)-2-[叔丁基(二甲基)硅基]氧基-2-(8-羟基-2-氧代-1H-喹啉-5-基)乙基]胺基]乙基-3-[2-[3-[[4-(4-甲氧基嘧啶基-2-基)-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-基]甲基]苯基]乙氧基]-N-甲基-丙酰胺(30G)
N-[2-[[(2R)-2-(tert-butyl(dimethyl)silyl)oxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]-3-[2-[3-[[4-(4-methoxpyrimidin-2-yl)-1-oxa-4,9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]-N-methyl-propanamide
将30F(0.500g,0.875mmol)溶于THF(10mL)中,加入TsOH·H2O(0.832g,4.37mmol),40℃搅拌1小时后,滴加饱和碳酸氢钠水溶液(50mL)淬灭反应,用DCM(100mL×2)萃取,合并后的有机相用无水硫酸钠干燥,减压浓缩后得到的粗产品溶于DCM(10mL)和甲醇(3mL)的混合溶剂中,加入5-[(1R)-2-氨基-1-[叔丁基(二甲基)甲硅烷基]氧基甲基]-8-羟基-1H-喹啉-2-酮(0.267g,0.80mmol),室温搅拌30分钟后加入三乙酰氧基硼氢化钠(0.56g,2.62mmol),室温搅拌3小时。加入饱和碳酸氢钠水溶液(50mL),用DCM(100mL×2)萃取,合并后的有机相用无水硫酸钠干燥,减压浓缩后柱层析分离(DCM:甲醇(v/v)=1:0~8:1)得到黄色固体状的30G(0.16g,产率21%)。
LCMS m/z=422.8[M/2+1]。
第七步:N-[2-[[(2R)-2-羟基-2-(8-羟基-2-氧代-1H-喹啉-5-基)乙基]胺基]乙基]-3-[2-[3-[[4-(4-甲氧基嘧
啶基-2-基)-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-基]甲基]苯基]乙氧基]-N-甲基-丙酰胺(化合物30)
N-[2-[[(2R)-2-hydroxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]-3-[2-[3-[[4-(4-methoxypyrimidin-2-yl)-1-oxa-4,9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]-N-methyl-propanamide
将30G(0.16g,0.19mmol)溶于THF(10mL)中,加入三乙胺三氢氟酸(0.061g,0.38mmol),室温搅拌12小时。依次滴加饱和碳酸氢钠水溶液(50mL),甲醇(5mL)淬灭反应,用DCM(100mL×2)萃取,合并后的有机相用无水硫酸钠干燥,减压浓缩后柱层析分离(DCM:甲醇(v/v)=1:0~8:1)得到黄色固体状的化合物30(0.06g,产率40%)。
1H NMR(400MHz,CD3OD)δ8.20(dd,1H),7.87(d,1H),7.08(dq,5H),6.92–6.82(m,1H),6.52(dd,1H),5.89(d,1H),5.18–5.01(m,1H),3.74(s,3H),3.68–3.49(m,10H),3.49–3.40(m,3H),3.37(dd,1H),3.02–2.89(m,3H),2.85–2.76(m,3H),2.72(dd,3H),2.54–2.43(m,4H),2.39(t,2H),1.72(d,2H),1.53(t,2H).
LCMS m/z=731.2[M+1]。
实施例31:N-[2-[[(2R)-2-羟基-2-(8-羟基-2-氧代-1H-喹啉-5-基)乙基]胺基]乙基]-N-甲基-3-[2-[3-[[4-(4-三氟甲基嘧啶基-2-基)-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-基]甲基]苯基]乙氧基]丙酰胺(化合物31)
N-[2-[[(2R)-2-hydroxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]-N-methyl-3-[2-[3-[[4-[4-(trifluoromethyl)pyrimidin-2-yl]-1-oxa-4,9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]propanamide
第一步:4-(4-三氟甲基嘧啶-2-基)-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-碳酸叔丁酯(31B)
tert-butyl 4-[4-(trifluoromethyl)pyrimidin-2-yl]-1-oxa-4,9-diazaspiro[5.5]undecane-9-carboxy late
将1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-羧酸叔丁酯(0.77g,3.0mmol)置于DMF(20mL)中,依次加入2-溴-3-三氟甲基嘧啶(31A)(0.55g,3.0mmol)、碳酸钾(0.83g,6.0mmol),升温50℃搅拌4小时。加入乙酸乙酯(200mL),水(100mL),萃取,有机相和水相分离,水相用乙酸乙酯(100mL×1)萃取,合并后的有机相用饱和食盐水洗(100mL),无水硫酸钠干燥,减压浓缩后柱层析分离(石油醚/乙酸乙酯(v/v)
=1:1)得到黄色液体状的31B(1.2g,产率100%)。
LCMS m/z=425.2[M+23]。
第二步:4-(4-三氟甲基嘧啶-2-基)-1-氧杂-4,9-二氮杂螺[5.5]十一烷(31C)
4-[4-(trifluoromethyl)pyrimidin-2-yl]-1-oxa-4,9-diazaspiro[5.5]undecane
将31B(1.61g,4.0mmol)溶于DCM(20mL)中,加入三氟乙酸(8mL),室温搅拌2小时。减压浓缩除去反应溶剂,向残余物中加入甲苯(5mL),减压浓缩后得到黄色液体状的2-(4-三氟甲基嘧啶-2-基)-11-氧杂-2,9-二氮杂螺[5.5]十一烷(31C)的三氟乙酸盐(1.21g,产率100%)。
LCMS m/z=303.2[M+1]。
第三步:3-[2-[3-[[4-(4-三氟甲基嘧啶-2-基)-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-基]甲基]苯基]乙氧基]丙酸叔丁酯(31D)
tert-butyl 3-[2-[3-[[4-(4-trifluoromethylpyrimidin-2-yl)-1-oxa-4,9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]propanoate
将31C的三氟乙酸盐(1.21g,4.0mmol)置于乙腈(25mL)中,依次加入中间体2(1.20g,4.0mmol),三乙胺(1.221g,12.07mmol),水(0.5mL),60℃搅拌24小时。待反应冷至室温后,加入水(50mL),用乙酸乙酯(100mL×2)萃取,合并后的有机相用无水硫酸钠干燥,减压浓缩后柱层析分离(DCM/甲醇(v/v)=15:1)得到黄色液体状的31D(1.90g,产率84.1%)。
LCMS m/z=565.5[M+1]。
第四步:3-[2-[3-[[4-(4-三氟甲基嘧啶基-2-基)1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-基]甲基]苯基]乙氧基]丙酸(31E)
3-[2-[3-[[4-(4-trifluoromethylpyrimidin-2-yl)-1-oxa-4,9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]propanoic acid
将31D(1.88g,3.33mmol)溶于DCM(15mL)中。室温,向反应滴加三氟乙酸(5mL),搅拌4小时。减压浓缩除去反应溶剂,向残余物中加入甲苯(10mL),减压浓缩后得到黄色液体状的31E的三氟乙酸(1.69g,产率100%)。
LCMS m/z=509.3[M+1]。
第五步:N-(2,2-二甲氧乙基)-N-甲基-3-[2-[3-[[4-(4-三氟甲基嘧啶-2-基)-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-基]甲基]苯基]乙氧基]-丙酰胺(31F)
N-(2,2-dimethoxyethyl)-N-methyl-3-[2-[3-[[4-(4-trifluoromethylpyrimidin-2-yl)-1-oxa-4,9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]propanamide
将31E的三氟乙酸盐(1.69g,3.33mmol)溶于DCM(20mL)中,依次加入三乙胺(0.674g,6.66mmol),甲氨基乙醛缩二甲醇(0.480g,4.00mmol),HATU(1.90g,5.00mmol),室温搅拌2小时。滴加饱和碳酸氢钠水溶液(50mL),用DCM(100mL×2)萃取,合并后的有机相用饱和食盐水(50mL)洗涤,无水硫酸钠干燥,减压浓缩后柱层析分离(DCM/甲醇(v/v)=15:1)得到黄色液体状的31F(2.03g,产率100%)。
LCMS m/z=610.4[M+1]。
第六步:N-[2-[[(2R)-2-[叔丁基(二甲基)硅基]氧基-2-(8-羟基-2-氧代-1H-喹啉-5-基)乙基]胺基]乙基-N-甲基-3-[2-[3-[[4-(4-三氟甲基嘧啶-2-基)-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-基]甲基]苯基]乙氧基]丙酰胺(31G)
N-[2-[[(2R)-2-(tert-butyl(dimethyl)silyl)oxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]-N-methyl-3-[2-[3-[[4-(4-trifluoromethylpyrimidin-2-yl)-1-oxa-4,9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]propanamide
将31F(0.500g,0.82mmol)溶于THF(10mL)中,加入TsOH·H2O(0.78g,4.10mmol),40℃搅拌1小时后,滴加饱和碳酸氢钠水溶液(50mL)淬灭反应,用DCM(100mL×2)萃取,合并后的有机相用无水硫酸钠干燥,减压浓缩后得到的粗产品溶于DCM(10mL)和甲醇(3mL)的混合溶剂中,加入5-[(1R)-2-氨基-1-[叔丁基(二甲基)甲硅烷基]氧基甲基]-8-羟基-1H-喹啉-2-酮(0.267g,0.80mmol),室温搅拌30分钟后加入三乙酰氧基硼氢化钠(0.53g,2.46mmol),室温搅拌3小时。加入饱和碳酸氢钠水溶液(50mL),用DCM(100mL×2)萃取,合并后的有机相用无水硫酸钠干燥,减压浓缩后柱层析分离(DCM:甲醇(v/v)=1:0~8:1)得到黄色固体状的31G(0.15g,产率20.6%)。
LCMS m/z=883.5[M+1]。
第七步:N-[2-[[(2R)-2-羟基-2-(8-羟基-2-氧代-1H-喹啉-5-基)乙基]胺基]乙基]-N-甲基-3-[2-[3-[[4-(4-三氟甲基嘧啶基-2-基)-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-基]甲基]苯基]乙氧基]丙酰胺(化合物31)
N-[2-[[(2R)-2-hydroxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]-N-methyl-3-[2-[3-[[4-[4-(trifluoromethyl)pyrimidin-2-yl]-1-oxa-4,9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]propanamide
将31G(0.15g,0.17mmol)溶于THF(10mL)中,加入三乙胺三氢氟酸(0.055g,0.34mmol),室温搅拌12小时。依次滴加饱和碳酸氢钠水溶液(50mL),甲醇(5mL)淬灭反应,用DCM(100mL×2)萃取,合并后的有机相用无水硫酸钠干燥,减压浓缩后柱层析分离(DCM:甲醇(v/v)=1:0~8:1)得到黄色固体状的化合物31(0.07g,产率50%)。
1H NMR(400MHz,CD3OD)δ8.55(d,1H),8.36(t,1H),7.27–7.07(m,5H),7.03–6.94(m,1H),6.85(d,1H),6.74–6.56(m,1H),5.28–5.11(m,1H),3.90–3.79(m,2H),3.78–3.69(m,6H),3.69–3.62(m,2H),3.60–3.51(m,3H),3.51–3.43(m,1H),3.03(s,2H),2.96(d,1H),2.90(s,2H),2.86–2.75(m,3H),2.60(dd,4H),2.50(t,2H),1.92(d,1H),1.83(d,2H),1.64(dd,2H).
LCMS m/z=384.7[M/2+1].
实施例32:3-[2-[3-[[4-[1-(环丙基甲基)吡唑-3-羰基]-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-基]甲基]苯基]乙氧基]-N-[2-[[(2R)-2-羟基-2-(8-羟基-2-氧代-1H-喹啉-5-基)乙基]氨基]乙基]-N-异丙基-丙酰胺(化合物32)
3-[2-[3-[[4-[1-(cyclopropylmethyl)pyrazole-3-carbonyl]-1-oxa-4,9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]-N-[2-[[(2R)-2-hydroxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]-N-isopropyl-propanamide
第一步:3-[2-[3-[[4-[1-(环丙甲基)吡唑-3-羰基]-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-基]甲基]苯基]乙氧基]-N-(2,2-二甲氧乙基)-N-异丙基-丙酰胺(32A)
3-[2-[3-[[4-[1-(cyclopropylmethyl)pyrazole-3-carbonyl]-1-oxa-4,9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]-N-(2,2-dimethoxyethyl)-N-isopropyl-propanamide
将22A(0.81g,1.4mmol)置加入DCM(10mL),0℃滴加三氟乙酸(5mL),滴毕,室温反应1小时,将体系减压浓缩,加入甲苯(20mL)后再次减压浓缩得到残留物。在0℃和氮气条件下向残留物中依次加入DCM(15mL)和三乙胺(0.70g,7.0mmol),搅拌均匀后加入N-(2,2-二甲氧基乙基)丙-2-胺(参考CN102625808制备得到)(0.31g,2.1mmol)和HATU(0.79g,2.1mmol),升至室温反应1小时。反应结束后加入水(30mL),DCM(30mL×2)萃取,合并有机相,饱和食盐水(40mL)洗涤,无水硫酸钠干燥,过滤,滤液减压浓缩,残留物经硅胶柱层析纯化(DCM/甲醇(v/v)=1:20)得到黄色液体状的32A(0.85g,产率:96%)。
LCMS m/z=640.4[M+1]
第二步:N-[2-[[(2R)-2-[叔丁基(二甲基)硅基]氧基-2-(8-羟基-2-氧代-1H-喹啉-5-基)乙基]氨基]乙基]-3-[2-[3-[[4-[1-(环丙基甲基)吡唑-3-羰基]-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-基]甲基]苯基]乙氧基]-N-异丙基-丙酰胺(32B)
N-[2-[[(2R)-2-[tert-butyl(dimethyl)silyl]oxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]-3-[2-[3-[[4-[1-(cyclopropylmethyl)pyrazole-3-carbonyl]-1-oxa-4,9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]-N-isopropyl-propanamide
将32A(0.85g,1.3mmol)溶于THF(15mL)中,加入TsOH·H2O(0.97g,5.1mmol),40℃搅拌1小时后,滴加饱和碳酸氢钠水溶液(50mL)淬灭反应,用DCM(100mL×2)萃取,合并后的有机相用无水硫酸钠干燥,减压浓缩后得到的粗产品溶于DCM(10mL)和甲醇(3mL)的混合溶剂中,加入5-[(1R)-2-氨基-1-[叔丁基(二甲基)甲硅烷基]氧基甲基]-8-羟基-1H-喹啉-2-酮(0.34g,1.0mmol),室温搅拌30分钟后加入三乙酰氧基硼氢化钠(0.65g,3.0mmol),室温搅拌3小时。加入饱和碳酸氢钠水溶液(50mL),用DCM(100mL×2)萃取,合并后的有机相用无水硫酸钠干燥,减压浓缩后柱层析分离(DCM:甲醇(v/v)=1:0~8:1)得到黄色固体状的32B(0.31g,产率33%)。
LCMS m/z=456.9[M/2+1]。
第三步:3-[2-[3-[[4-[1-(环丙基甲基)吡唑-3-羰基]-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-基]甲基]苯基]乙氧基]-N-[2-[[(2R)-2-羟基-2-(8-羟基-2-氧代-1H-喹啉-5-基)乙基]氨基]乙基]-N-异丙基-丙酰胺(化合物32)
3-[2-[3-[[4-[1-(cyclopropylmethyl)pyrazole-3-carbonyl]-1-oxa-4,9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]-N-[2-[[(2R)-2-hydroxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]-N-isopropyl-propanamide
将32B(0.32g,0.35mmol)溶于THF(10mL)中,加入三乙胺三氢氟酸(0.99g,6.1mmol),室温搅拌12小时。依次滴加饱和碳酸氢钠水溶液(50mL),甲醇(5mL)淬灭反应,用DCM(100mL×2)萃取,合并后的有机相用无水硫酸钠干燥,减压浓缩后柱层析分离(DCM:甲醇(v/v)=1:0~8:1)得到黄色固体状的化合物32(0.13g,产率48%)。
1H NMR(400MHz,DMSO-d6)δ10.23(br,1H),8.19(dd,1H),7.81(d,1H),7.18(d,1H),7.13–7.03(m,4H),6.91(d,1H),6.55(s,1H),6.48(d,1H),5.00(dd,1H),4.04-3.99(m,2H),3.96-3.86(m,2H),3.66–3.49(m,9H),3.43-3.22(d,8H),3.19–3.13(m,2H),2.80–2.67(m,4H),2.65-2.57(m,2H),2.39-2.25(m,4H),1.67(d,2H),1.56-1.44(s,2H),1.08–1.06(m,2H),1.02(d,2H),0.57-0.49(m,2H),0.40-0.32(m,2H).
LCMS m/z=399.8[M/2+1].
实施例33:3-[2-[2-氯-3-[[4-[1-(环丙基甲基)吡唑-3-羰基]-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-基]甲基]苯基]乙氧基]-N-[2-[[(2R)-2-羟基-2-(8-羟基-2-氧代-1H-喹啉-5-基)乙基]氨基]乙基]-N-甲基-丙酰胺(化合物33)
3-[2-[2-chloro-3-[[4-[1-(cyclopropylmethyl)pyrazole-3-carbonyl]-1-oxa-4,9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]-N-[2-[[(2R)-2-hydroxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]-N-methyl-propanamide
第一步:[9-[[2-氯-3-(2-羟乙基)苯基]甲基]-1-氧杂-4,9-二氮杂螺[5.5]十一烷-4-基]-[1-(环丙甲基)吡唑-3-基]甲酮(33A)
[9-[[2-chloro-3-(2-hydroxyethyl)phenyl]methyl]-1-oxa-4,9-diazaspiro[5.5]undecan-4-yl]-[1-(cyclopropylmethyl)pyrazol-3-yl]methanone
将1E(1.48g,4.86mmol)溶于THF(30mL)中,加入2-氯-3-(2-羟乙基)苯甲醛(0.9g,5.0mmol),室温搅拌1小时后,加入三乙酰氧基硼氢化钠(2.85g,13.4mmol),室温搅拌3小时。加入饱和碳酸氢钠水溶液(50mL),用DCM(100mL×2)萃取,合并后的有机相用无水硫酸钠干燥,减压浓缩后柱层析分离(DCM:甲醇(v/v)=1:0~8:1)得到黄色液体状的33A(1.3g,产率57%)。
第二步:3-[2-[2-氯-3-[[4-[1-(环丙甲基)吡唑-3-羰基]-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-基]甲基]苯基]乙氧基]丙酸叔丁酯(33B)
tert-butyl 3-[2-[2-chloro-3-[[4-[1-(cyclopropylmethyl)pyrazole-3-carbonyl]-1-oxa-4,9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]propanoate
将33A(1.3g,2.7mmol)溶于乙腈(30mL)中,依次加入丙烯酸叔丁酯(1.38g,10.0mmol),苄基三甲基氢氧化铵(0.50g,1.2mmol,40%的甲醇溶液),室温搅拌3小时。减压浓缩除去大部分反应溶剂,柱层析分离(石油醚/乙酸乙酯(v/v)=10:1)得到黄色液体状的33B(1.5g,产率91%)。
第三步:3-[2-[2-氯-3-[[4-[1-(环丙甲基)吡唑-3-羰基]-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-基]甲基]苯基]乙氧基]-N-(2,2-二甲氧基乙基)-N-甲基-丙酰胺(33C)
3-[2-[2-chloro-3-[[4-[1-(cyclopropylmethyl)pyrazole-3-carbonyl]-1-oxa-4,9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]-N-(2,2-dimethoxyethyl)-N-methyl-propanamide
将33B(1.5g,2.5mmol)加入DCM(10mL),0℃滴加三氟乙酸(5mL),滴毕,室温反应1小时后,反应液减压浓缩,加入甲苯(20mL)后再次减压浓缩得到残留物。在0℃和氮气条件下向残留物中依次加入DCM(15mL)和三乙胺(1.0g,9.9mmol),搅拌均匀后加入N-(2,2-二甲氧基乙基)甲-1-氨(0.5g,4.2mmol)和HATU(1.8g,4.7mmol),室温反应1小时。反应结束后加入水(30mL),用DCM(30mL×2)萃取,合并有机相,饱和食盐水(40mL)洗涤,无水硫酸钠干燥,过滤,滤液减压浓缩,残留物经硅胶柱层析纯化(DCM/甲醇(v/v)=1:20)得到黄色液体状的33C(1.59g,产率98%)。
第四步:N-[2-[[(2R)-2-[叔丁基(二甲基)硅基]氧基-2-(8-羟基-2-氧代-1H-喹啉-5-基)乙基]氨基]乙基]-3-[2-[2-氯-3-[[4-[1-(环丙基甲基)吡唑-3-羰基]-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-基]甲基]苯基]乙氧基]-N-甲基-丙酰胺(33D)
N-[2-[[(2R)-2-[tert-butyl(dimethyl)silyl]oxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]-3-[2-[2-chloro-3-[[4-[1-(cyclopropylmethyl)pyrazole-3-carbonyl]-1-oxa-4,9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]-N-methyl-propanamide
将33C(0.80g,1.2mmol)溶于THF(15mL)中,加入TsOH·H2O(1.2g,6.2mmol),40℃搅拌1小时后,滴加饱和碳酸氢钠水溶液(50mL)淬灭反应,用DCM(100mL×2)萃取,合并后的有机相用无水硫酸钠干燥,减压浓缩后得到的粗产品溶于DCM(10mL)和甲醇(3mL)的混合溶剂中,加入5-[(1R)-2-氨基-1-[叔丁基(二甲基)甲硅烷基]氧基甲基]-8-羟基-1H-喹啉-2-酮(0.33g,1.0mmol),室温搅拌30分钟后加入三乙酰氧基硼氢化钠(0.79g,3.7mmol),室温搅拌3小时。加入饱和碳酸氢钠水溶液(50mL),用DCM(100mL×2)萃取,合并后的有机相用无水硫酸钠干燥,减压浓缩后柱层析分离(DCM:甲醇(v/v)=1:0~8:1)得到黄色固体状的33D(0.16g,产率14%)。
第五步:3-[2-[2-氯-3-[[4-[1-(环丙基甲基)吡唑-3-羰基]-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-基]甲基]苯基]乙氧基]-N-[2-[[(2R)-2-羟基-2-(8-羟基-2-氧代-1H-喹啉-5-基)乙基]氨基]乙基]-N-甲基-丙酰胺(化合物33)
3-[2-[2-chloro-3-[[4-[1-(cyclopropylmethyl)pyrazole-3-carbonyl]-1-oxa-4,9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]-N-[2-[[(2R)-2-hydroxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]-N-methyl-propanamide
将33D(0.16g,0.17mmol)溶于THF(10mL)中,加入三乙胺三氢氟酸(0.8g,4.9mmol),室温搅拌12小时。依次滴加饱和碳酸氢钠水溶液(50mL),甲醇(5mL)淬灭反应,用DCM(100mL×2)萃取,合并后的有机相用无水硫酸钠干燥,减压浓缩后柱层析分离(DCM:甲醇(v/v)=1:0~8:1)得到黄色固体状的化合物33(0.08g,产率60%)。
1H NMR(400MHz,DMSO-d6)δ10.20(br,1H),8.20(t,1H),7.82(d,1H),7.36-7.28(s,1H),7.26-7.18(d,2H),7.08(dd,1H),6.96(dd,1H),6.56(s,1H),6.49(dd,1H),5.14–5.04(m,1H),4.05-3.90(m,5H),3.67–3.48(m,8H),3.37(dd,4H),2.93(s,3H),2.79-2.70(m,9H),2.44–2.32(m,4H),1.72-1.65(m,2H),1.58-1.48(m,2H),0.56-0.50(m,2H),0.40-0.34(m,2H).
LCMS m/z=402.8[M/2+1].
实施例34:N-丁基-N-[2-[[(2R)-2-羟基-2-(8-羟基-2-氧代-1H-喹啉-5-基)乙基]氨基]乙基]-3-[2-[3-[[4-(4-甲基-2-吡啶基)-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-基]甲基]苯基]乙氧基]丙酰胺(化合物34)
N-butyl-N-[2-[[(2R)-2-hydroxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]-3-[2-[3-[[4-(4-methyl-2-pyridyl)-1-oxa-4,9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]propanamide
第一步:N-丁基-N-(2,2-二甲氧基乙基)-3-[2-[3-[[4-(4-甲基-2-吡啶基)-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-基]甲基]苯基]乙氧基]丙酰胺(34A)
N-butyl-N-(2,2-dimethoxyethyl)-3-[2-[3-[[4-(4-methyl-2-pyridyl)-1-oxa-4,9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]propanamide
将23C(0.42g,0.82mmol)加入DCM(10mL)中,0℃滴加三氟乙酸(2mL),滴毕,室温反应1小时后,反应液减压浓缩,加入甲苯(20mL)后再次减压浓缩得到残留物。在0℃和氮气条件下向残留物中依次加入DCM(15mL)和三乙胺(0.40g,3.9mmol),搅拌均匀后加入N-(2,2-二甲氧基乙基)丁-2-胺(参考CN
102625808制备得到)(0.2g,1.2mmol)和HATU(0.50g,1.3mmol),室温反应1小时。加入水(30mL),用DCM(30mL×2)萃取,合并有机相,饱和食盐水(40mL)洗涤,无水硫酸钠干燥,过滤,滤液减压浓缩,残留物经硅胶柱层析纯化(DCM/甲醇(v/v)=1:20)得到黄色液体状的34A(0.42g,产率85%)。
第二步:N-丁基-N-[2-[[(2R)-2-[叔丁基(二甲基)硅基]氧基-2-(8-羟基-2-氧代-1H-喹啉-5-基)乙基]氨基]乙基]-3-[2-[3-[[4-(4-甲基-2-吡啶基)-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-基]甲基]苯基]乙氧基]丙酰胺(34B)
N-butyl-N-[2-[[(2R)-2-[tert-butyl(dimethyl)silyl]oxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]-3-[2-[3-[[4-(4-methyl-2-pyridyl)-1-oxa-4,9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]propanamide
将34A(0.20g,0.34mmol)溶于THF(15mL)中,加入TsOH·H2O(0.32g,1.7mmol),40℃搅拌1小时,滴加饱和碳酸氢钠水溶液(50mL)淬灭反应,用DCM(100mL×2)萃取,合并后的有机相用无水硫酸钠干燥,减压浓缩后得到的粗产品溶于DCM(10mL)和甲醇(3mL)的混合溶剂中,加入5-[(1R)-2-氨基-1-[叔丁基(二甲基)甲硅烷基]氧基甲基]-8-羟基-1H-喹啉-2-酮(0.09g,0.27mmol),室温搅拌30分钟后加入三乙酰氧基硼氢化钠(0.21g,1.0mmol),室温搅拌3小时。加入饱和碳酸氢钠水溶液(50mL),用DCM(100mL×2)萃取,合并后的有机相用无水硫酸钠干燥,减压浓缩后柱层析分离(DCM:甲醇(v/v)=1:0~8:1)得到黄色固体状的34B(0.11g,产率38%)
第三步:N-丁基-N-[2-[[(2R)-2-羟基-2-(8-羟基-2-氧代-1H-喹啉-5-基)乙基]氨基]乙基]-3-[2-[3-[[4-(4-甲基-2-吡啶基)-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-基]甲基]苯基]乙氧基]丙酰胺(化合物34)
N-butyl-N-[2-[[(2R)-2-hydroxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]-3-[2-[3-[[4-(4-methyl-2-pyridyl)-1-oxa-4,9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]propanamide
将34B(0.11g,0.13mmol)溶于THF(10mL)中,加入三乙胺三氢氟酸(0.99g,6.1mmol),室温搅拌12小时。依次滴加饱和碳酸氢钠水溶液(50mL),甲醇(5mL)淬灭反应,用DCM(100mL×2)萃取,合并后的有机相用无水硫酸钠干燥,减压浓缩后柱层析分离(DCM:甲醇(v/v)=1:0~8:1)得到黄色固体状的化合物34(0.016g,产率17%)。
1H NMR(400MHz,DMSO-d6)δ10.24(br,1H),8.18(d,1H),7.93(d,1H),7.23–7.16(m,1H),7.14–7.04(m,4H),6.91(d,1H),6.65(s,1H),6.51-6.43(m,2H),5.10-4.87(m,1H),3.70–3.64(m,2H),3.61–3.54(m,4H),3.45–3.14(m,17H),2.79-2.60(m,6H),2.43-2.38(m,2H),2.32-2.24(m,2H),2.20(s,3H),1.87–1.64(m,3H),1.63-1.47(m,3H).
LCMS m/z=378.3[M/2+1]。
实施例35:N-丁基-N-[2-[2-(5-羟基-3-氧-4H-1,4-苯并噁嗪-8-基)乙基氨基]乙基]-3-[2-[3-[[4-(4-甲基-2-吡啶基)-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-基]甲基]苯基]乙氧基]丙酰胺(化合物35)
N-butyl-N-[2-[2-(5-hydroxy-3-oxo-4H-1,4-benzoxazin-8-yl)ethylamino]ethyl]-3-[2-[3-[[4-(4-methyl-2-pyridyl)-1-oxa-4,9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]propanamide
将34A(0.21g,0.35mmol)溶于N-甲基吡咯烷酮(15mL)中,加入8-(2-胺乙基)-5-羟基-4H-1,4-苯并噁嗪-3-酮的乙酸盐(0.094g,0.35mmol),乙酸(0.021g,0.35mmol)。室温搅拌30分钟后,加入三乙酰氧基硼氢化钠(0.22g,1.1mmol),5小时后终止反应。减压浓缩除去大部分反应溶剂,向残余物中缓慢滴加饱和碳酸氢钠水溶液(50mL),用DCM萃取(50mL×2),合并后的有机相用饱和食盐水洗(50mL),无水硫酸钠干燥后,减压浓缩柱层析分离[(DCM/甲醇(v/v)=15:1)]得到白色固体状的化合物35(0.05g,产率20%)。
1H NMR(400MHz,DMSO-d6)δ9.92(br,1H),7.94(d,1H),7.30–7.05(m,5H),6.65(d,2H),6.51–6.44(m,2H),4.54–4.42(m,2H),3.67(d,3H),3.64–3.17(m,25H),2.78(dd,4H),2.67(d,1H),2.39(d,2H),1.77(t,2H),1.64–1.52(m,3H),1.51–1.37(m,2H).
LCMS m/z=372.3[M/2+1].
实施例36:N-环戊基-N-[2-[[(2R)-2-羟基-2-(8-羟基-2-氧代-1H-喹啉-5-基)乙基]氨基]乙基]-3-[2-[3-[[4-(4-甲基-2-吡啶基)-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-基]甲基]苯基]乙氧基]丙酰胺(化合物36)
N-cyclopentyl-N-[2-[[(2R)-2-hydroxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]-3-[2-[3-[[4-(4-methyl-2-pyridyl)-1-oxa-4,9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]propanamide
第一步:N-环戊基-N-(2,2-二甲氧基乙基)-3-[2-[3-[[4-(4-甲基-2-吡啶基)-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-基]甲基]苯基]乙氧基]丙酰胺(36A)
N-cyclopentyl-N-(2,2-dimethoxyethyl)-3-[2-[3-[[4-(4-methyl-2-pyridyl)-1-oxa-4,9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]propanamide
将23C(0.42g,0.82mmol)加入DCM(10mL)中,0℃滴加三氟乙酸(2mL),滴毕,室温反应1小时后,反应液减压浓缩,加入甲苯(20mL)后再次减压浓缩得到残留物。在0℃和氮气条件下向残留物中依次加入DCM(15mL)和三乙胺(0.40g,3.9mmol),搅拌均匀后加入N-(2,2-二甲氧基乙基)环戊胺(参考CN102625808制备得到)(0.20g,0.23mmol)和HATU(0.50g,1.3mmol),室温反应1小时。反应结束后加入水(30mL),用DCM(30mL×2)萃取,合并有机相,饱和食盐水(40mL)洗涤,无水硫酸钠干燥,过滤,滤液减压浓缩,残留物经硅胶柱层析纯化(DCM/甲醇(v/v)=1:20)得到黄色液体状的36A(0.41g,产率80%)。
第二步:N-[2-[[(2R)-2-[叔丁基(二甲基)硅基]氧基-2-(8-羟基-2-氧代-1H-喹啉-5-基)乙基]氨基]乙基]-N-环戊基-3-[2-[3-[[4-(4-甲基-2-吡啶基)-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-基]甲基]苯基]乙氧基]丙酰胺
(36B)
N-[2-[[(2R)-2-[tert-butyl(dimethyl)silyl]oxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]-N-cyclopentyl-3-[2-[3-[[4-(4-methyl-2-pyridyl)-1-oxa-4,9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]propanamide
将36A(0.21g,0.34mmol)溶于THF(15mL)中,加入TsOH·H2O(0.33g,1.7mmol),40℃搅拌1小时后,滴加饱和碳酸氢钠水溶液(50mL)淬灭反应,用DCM(100mL×2)萃取,合并后的有机相用无水硫酸钠干燥,减压浓缩后得到的粗产品溶于DCM(10mL)和甲醇(3mL)的混合溶剂中,加入5-[(1R)-2-氨基-1-[叔丁基(二甲基)甲硅烷基]氧基甲基]-8-羟基-1H-喹啉-2-酮(0.12g,0.34mmol),室温搅拌30分钟后加入三乙酰氧基硼氢化钠(0.22g,1.0mmol),室温搅拌3小时。加入饱和碳酸氢钠水溶液(50mL),用DCM(100mL×2)萃取,合并后的有机相用无水硫酸钠干燥,减压浓缩后柱层析分离(DCM:甲醇(v/v)=1:0~8:1)得到黄色固体状的36B(0.09g,产率30%)。
第三步:N-环戊基-N-[2-[[(2R)-2-羟基-2-(8-羟基-2-氧代-1H-喹啉-5-基)乙基]氨基]乙基]-3-[2-[3-[[4-(4-甲基-2-吡啶基)-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-基]甲基]苯基]乙氧基]丙酰胺(化合物36)
N-cyclopentyl-N-[2-[[(2R)-2-hydroxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]-3-[2-[3-[[4-(4-methyl-2-pyridyl)-1-oxa-4,9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]propanamide
将36B(0.09g,0.10mmol)溶于THF(10mL)中,加入三乙胺三氢氟酸(0.69g,4.3mmol),室温搅拌12小时。依次滴加饱和碳酸氢钠水溶液(50mL),甲醇(5mL)淬灭反应,用DCM(100mL×2)萃取,合并后的有机相用无水硫酸钠干燥,减压浓缩后柱层析分离(DCM:甲醇(v/v)=1:0~8:1)得到黄色固体状的化合物36(0.02g,产率30%)。
1H NMR(400MHz,DMSO-d6)δ10.24(br,1H),8.19(d,1H),7.93(d,1H),7.21–7.16(m,1H),7.14–7.05(m,4H),6.91(d,1H),6.65(s,1H),6.50-6.44(m,2H),5.03–4.96(m,1H),3.69–3.65(m,2H),3.61-3.54(m,4H),3.44–3.21(m,10H),2.77-2.73(m,2H),2.71-2.56(dd,4H),2.44–2.39(m,2H),2.30-2.24(m,2H),2.20(s,3H),1.76-1.70(m,3H),1.65–1.40(m,10H).
LCMS m/z=384.2[M/2+1].
实施例37:N-[2-[[(2R)-2-羟基-2-(8-羟基-2-氧代-1H-喹啉-5-基)乙基]胺基]乙基]-N-甲基-3-[2-[3-[[4-(2-丙炔基噻唑-4-羰基)-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-基]甲基]苯基]乙氧基]丙酰胺(化合物37)
N-[2-[[(2R)-2-hydroxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]-N-methyl-3-[2-[3-[[4-(2-prop-1-ynylthiazole-4-carbonyl)-1-oxa-4,9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]propanamide
第一步:3-[2-[3-[[4-(2-丙炔基噻唑-4-羰基)-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-基]甲基]苯基]乙氧基]丙酸叔丁酯(37A)
tert-butyl 3-[2-[3-[[4-(2-prop-1-ynylthiazole-4-carbonyl)-1-oxa-4,9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]propanoate
将5E的三氟乙酸盐(0.50g,1.2mmol)置于乙腈(15mL)中,加入中间体2(0.43g,1.2mmol),碳酸钾(0.86g,6.2mmol),水(0.5mL),室温搅拌8小时。加入水(50mL),用乙酸乙酯(100mL×2)萃取,合并后的有机相用无水硫酸钠干燥,减压浓缩后柱层析分离(DCM/甲醇(v/v)=15:1)得到黄色固体状的37A(0.60g,产率80%)。
LCMS m/z=568.3[M+1].
第二步:3-[2-[3-[[4-(2-丙炔噻唑-4-羰基)-11-氧杂-3,8-二氮杂螺[5.5]十一烷-3-基]甲基]苯基]乙氧基]丙酸(37B)
3-[2-[3-[[4-(2-prop-1-ynylthiazole-4-carbonyl)-1-oxa-4,9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]propanoic acid
将37A(0.60g,1.0mmol)溶于DCM(15mL)中,滴加三氟乙酸(5mL),搅拌4小时。减压浓缩除去反应溶剂,向残余物中加入甲苯(10mL),减压浓缩后得到黄色液体状的37B的三氟乙酸盐(0.55g,产率100%)。
LCMS m/z=512.3[M+1]。
第三步:N-(2,2-二甲氧基乙基)-N-甲基-3-[2-[3-[[4-(2-丙炔噻唑-4-羰基)-11-氧杂-3,8-二氮杂螺[5.5]十一烷-3-基]甲基]苯基]乙氧基]丙酰胺(37C)
N-(2,2-dimethoxyethyl)-N-methyl-3-[2-[3-[[4-(2-prop-1-ynylthiazole-4-carbonyl)-1-oxa-4,9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]propanamide
将37B的三氟乙酸(0.55g,1.1mmol)加入DCM(20mL)中,加入三乙胺(0.54g,5.4mmol),甲氨基乙醛缩二甲醇(0.19g,1.6mmol),HATU(0.61g,1.6mmol),室温搅拌2小时。滴加饱和碳酸氢钠水溶液(50mL),用DCM(100mL×2)萃取,合并后的有机相用饱和食盐水(50mL)洗涤,无水硫酸钠干燥,减压浓缩后柱层析分离(DCM/甲醇(v/v)=15:1)得到黄色固体状的37C(0.50g,产率80.0%)。
第四步:N-[2-[[(2R)-2-[叔丁基(二甲基)硅基]氧基-2-(8-羟基-2-氧代-1H-喹啉-5-基)乙基]胺基]乙基]-N-甲基-3-[2-[3-[[4-(2-丙炔噻唑-4-羰基)-11-氧杂-3,8-二氮杂螺[5.5]十一烷-3-基]甲基]苯基]乙氧基]丙酰胺(37D)
N-[2-[[(2R)-2-[tert-butyl(dimethyl)silyl]oxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]-N-methyl-3-[2-[3-[[4-(2-prop-1-ynylthiazole-4-carbonyl)-1-oxa-4,9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]propanamide
将37C(0.50g,0.8mmol)溶于THF(10mL)中,加入TsOH·H2O(0.80g,4.0mmol),40℃搅拌1小时,
滴加饱和碳酸氢钠水溶液(50mL)淬灭反应,用DCM(100mL×2)萃取,合并后的有机相用无水硫酸钠干燥,减压浓缩后得到的粗产品溶于DCM(10mL)和甲醇(3mL)的混合溶剂中,加入5-[(1R)-2-氨基-1-[叔丁基(二甲基)甲硅烷基]氧基甲基]-8-羟基-1H-喹啉-2-酮(0.31g,0.92mmol),室温搅拌30分钟后加入三乙酰氧基硼氢化钠(0.5g,2.4mmol),室温搅拌3小时。向其中加入饱和碳酸氢钠水溶液(50mL),用DCM(100mL×2)萃取,合并后的有机相用无水硫酸钠干燥,减压浓缩后柱层析分离(DCM:甲醇(v/v)=1:0~8:1)得到黄色固体状的37D(0.3g,产率40%)。
LCMS m/z=443.4[M/2+1].
第五步:N-[2-[[(2R)-2-羟基-2-(8-羟基-2-氧代-1H-喹啉-5-基)乙基]胺基]乙基]-N-甲基-3-[2-[3-[[4-(2-丙炔基噻唑-4-羰基)-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-基]甲基]苯基]乙氧基]丙酰胺(化合物37)
N-[2-[[(2R)-2-hydroxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]-N-methyl-3-[2-[3-[[4-(2-prop-1-ynylthiazole-4-carbonyl)-1-oxa-4,9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]propanamide
将37D(0.30g,0.34mmol)溶于THF(10mL)中,加入三乙胺三氢氟酸(0.55g,3.4mmol),室温搅拌12小时。依次滴加饱和碳酸氢钠水溶液(50mL),甲醇(5mL)淬灭反应,用DCM(100mL×2)萃取,合并后的有机相用无水硫酸钠干燥,减压浓缩后柱层析分离(DCM:甲醇(v/v)=1:0~8:1)得到黄色固体状的化合物37(0.10g,产率38%)。
1H NMR(400MHz,CDCl3)δ8.03-7.89(m,2H),7.26-7.07(m,5H),6.76(br,1H),6.48-6.44(m,1H),5.30-5.27(m,1H),3.94-3.53(m,14H),3.10-2.59(m,14H),2.11-1.64(m,9H).
LCMS m/z=772.3[M+1].
实施例38:3-[2-[3-[[4-[1-(环丙甲基)吡唑-3-羰基]-1-氧杂-4,9-杂螺[5.5]十一烷-9-基]甲基]苯基]乙氧基]-N-[2-[[(2R)-2-羟基-2-(8-羟基-2-氧代-1H-喹啉-5-基)乙基]胺基]乙基]-N-甲基-丙酰胺(化合物38)
3-[2-[3-[[4-[1-(cyclopropylmethyl)pyrazole-3-carbonyl]-1-oxa-4,9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]-N-[2-[[(2R)-2-hydroxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]-N-methyl-propanamide
第一步:N-[2-[[(2R)-2-[叔丁基(二甲基)硅基]氧基-2-(8-羟基-2-氧代-1H-喹啉-5-基)乙基]氨基]乙基]-3-[2-[3-[[4-[1-(环丙基甲基)吡唑-3-羰基]-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-基]甲基]苯基]乙氧基]-N-甲基-丙酰胺(38A)
N-[2-[[(2R)-2-[tert-butyl(dimethyl)silyl]oxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]-3-[2-
[3-[[4-[1-(cyclopropylmethyl)pyrazole-3-carbonyl]-1-oxa-4,9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]-N-methyl-propanamide
将22B(0.50g,0.82mmol)溶于THF(15mL)中,加入TsOH·H2O(0.78g,4.1mmol),40℃搅拌1小时后,滴加饱和碳酸氢钠水溶液(50mL)淬灭反应,用DCM(100mL×2)萃取,合并后的有机相用无水硫酸钠干燥,减压浓缩后得到的粗产品溶于DCM(10mL)和甲醇(3mL)的混合溶剂中,加入5-[(1R)-2-氨基-1-[叔丁基(二甲基)甲硅烷基]氧基甲基]-8-羟基-1H-喹啉-2-酮(0.27g,0.82mmol),室温搅拌30分钟后加入三乙酰氧基硼氢化钠(0.35g,1.6mmol),室温搅拌3小时。加入饱和碳酸氢钠水溶液(50mL),用DCM(100mL×2)萃取,合并后的有机相用无水硫酸钠干燥,减压浓缩后柱层析分离(DCM:甲醇(v/v)=1:0~8:1)得到黄色固体状的38A(0.3g,产率42%)。
LCMS m/z=885.5[M+1].
第二步:3-[2-[3-[[4-[1-(环丙甲基)吡唑-3-羰基]-1-氧杂-4,9-杂螺[5.5]十一烷-9-基]甲基]苯基]乙氧基]-N-[2-[[(2R)-2-羟基-2-(8-羟基-2-氧代-1H-喹啉-5-基)乙基]胺基]乙基]-N-甲基-丙酰胺(化合物38)
3-[2-[3-[[4-[1-(cyclopropylmethyl)pyrazole-3-carbonyl]-1-oxa-4,9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]-N-[2-[[(2R)-2-hydroxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]-N-methyl-propanamide
将38A(0.25g,0.28mmol)溶于THF(10mL)中,加入三乙胺三氢氟酸(0.46g,2.8mmol),室温搅拌12小时。依次滴加饱和碳酸氢钠水溶液(50mL),甲醇(5mL)淬灭反应,用DCM(100mL×2)萃取,合并后的有机相用无水硫酸钠干燥,减压浓缩后柱层析分离(DCM:甲醇(v/v)=1:0~8:1)得到黄色固体状的化合物38(0.1g,产率50%)。
1H NMR(400MHz,CDCl3)δ8.03(br,1H),7.48-7.41(m,1H),7.23-7.05(m,5H),6.78-6.41(m,3H),5.14-5.04(m,1H),4.08–3.44(m,19H),3.01-2.40(m,15H),1.83-1.61(m,4H),0.65-0.56(m,2H),0.37-0.28(m,2H).
LCMS m/z=771.4[M+1]。
实施例39:N-[2-[[(2R)-2-羟基-2-(8-羟基-2-氧代-1H-喹啉-5-基)乙基]胺基]乙基]-3-[2-[3-[[4-(4-异丙基-2-吡啶基)-1-氧杂-4,9-杂螺二氮[5.5]十一烷-9-基]甲基]苯基]乙氧基]-N-甲基-丙酰胺(化合物39)
N-[2-[[(2R)-2-hydroxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]-3-[2-[3-[[4-(4-isopropyl-2-pyridyl)-1-oxa-4,9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]-N-methyl-propanamide
第一步:4-(4-异丙基-2-吡啶基)-1-氧杂-4,9-二氮杂螺环[5.5]十一烷-9-羧酸叔丁酯(39B)
tert-butyl 4-(4-isopropyl-2-pyridyl)-1-oxa-4,9-diazaspiro[5.5]undecane-9-carboxylate
将1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-羧酸叔丁酯(2.56g,10.0mmol)溶于1,4-二氧六环(50mL)中,依次加入2-氯-4-异丙基-吡啶(39A)(WO2008100426,1.87g,12.0mmol),叔丁醇钠(2.4g,25.0mmol),4,5-双二苯基膦-9,9-二甲基氧杂蒽(0.81g,1.4mmol),醋酸钯(0.16g,0.7mmol),升温至回流氮气氛反应8小时。待反应冷至室温后,加入乙酸乙酯(200mL)和水(100mL),萃取,有机相和水相分离,水相用乙酸乙酯(100mL)萃取,合并后的有机相用饱和食盐水洗(100mL),无水硫酸钠干燥,减压浓缩后柱层析分离(石油醚/乙酸乙酯(v/v)=1:1)得到黄色液体状的39B(2.5g,产率67%)。
第二步:4-(4-异丙基-2-吡啶)-1-氧杂-4,9-二氮杂螺环[5.5]十一烷(39C)
4-(4-isopropyl-2-pyridyl)-1-oxa-4,9-diazaspiro[5.5]undecane
称取39B(0.75g,2.0mmol),溶于DCM(10mL)中,加入三氟乙酸(3mL),室温搅拌2小时。减压浓缩除去反应溶剂,向残余物中加入甲苯(5mL),减压浓缩后得到黄色液体状的39C的三氟乙酸盐(0.80,产率100%)。
LCMS m/z=276.1[M+1].
第三步:3-[2-[3-[[4-(4-异丙基-2-吡啶基)-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-基]甲基]苯基]乙氧基]丙酸叔丁酯(39D)
tert-butyl 3-[2-[3-[[4-(4-isopropyl-2-pyridyl)-1-oxa-4,9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]propanoate
将39C的三氟乙酸盐(0.8g,2.0mmol)加入乙腈(15mL)中,加入3-[2-[3-(2-溴甲基)苯基]乙氧基]丙酸叔丁酯(中间体2)(0.71g,2.0mmol),碳酸钾(1.4g,10.0mmol),水(0.5mL),室温搅拌8小时。待反应冷至室温后,加入水(50mL),用乙酸乙酯(100mL×2)萃取,合并后的有机相用无水硫酸钠干燥,减压浓缩后柱层析分离(DCM/甲醇(v/v)=15:1)得到黄色液体状的39D(1.1g,产率100%)。
LCMS m/z=539.3[M+1]。
第四步:3-[2-[3-[[4-(4-异丙基-2-吡啶基)-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-基]甲基]苯基]乙氧基]丙酸(39E)
3-[2-[3-[[4-(4-isopropyl-2-pyridyl)-1-oxa-4,9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]propanoicacid
将39D(1.1g,2.0mmol)溶于DCM(15mL)中,滴加三氟乙酸(5mL),室温搅拌4小时。减压浓缩除去反应溶剂,向残余物中加入甲苯(10mL),减压浓缩后得到黄色液体状的39E(1.0g,产率100%)。
LCMS m/z=482.4[M+1]。
第五步:N-(2,2-二甲氧基乙基)-3-[2-[3-[[4-(4-异丙基-2-吡啶基)-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-基]甲基]苯基]乙氧基]-N-甲基-丙酰胺(39F)
N-(2,2-dimethoxyethyl)-3-[2-[3-[[4-(4-isopropyl-2-pyridyl)-1-oxa-4,9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]-N-methyl-propanamide
将39E(0.99g,2.0mmol)溶于DCM(20mL)中,依次加入三乙胺(1.0g,10.0mmol),甲氨基乙醛缩二甲醇(0.36g,3.0mmol),HATU(1.2g,3.0mmol),室温搅拌2小时。滴加饱和碳酸氢钠水溶液(50mL),用DCM(100mL×2)萃取,合并后的有机相用饱和食盐水(50mL)洗涤,无水硫酸钠干燥,减压浓缩后柱层析分离(DCM/甲醇(v/v)=15:1)得到黄色固体状的39F(0.81g,产率68%)。
LCMS m/z=584.3[M+1]。
第六步:N-[2-[[(2R)-2-[叔丁基(二甲基)硅基]氧基-2-(8-羟基-2-氧代-1H-喹啉-5-基)乙基]胺基]乙基]-3-[2-[3-[[4-(4-异丙基-2-吡啶基)-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-基]甲基]苯基]乙氧基]-N-甲基-丙酰胺(39G)
N-[2-[[(2R)-2-[tert-butyl(dimethyl)silyl]oxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]-3-[2-[3-[[4-(4-isopropyl-2-pyridyl)-1-oxa-4,9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]-N-methyl-propanamide
将39F(0.4g,0.7mmol)溶于THF(10mL)中,加入TsOH·H2O(0.7g,3.5mmol),40℃搅拌1小时后,滴加饱和碳酸氢钠水溶液(50mL)淬灭反应,用DCM(100mL×2)萃取,合并后的有机相用无水硫酸钠干燥,减压浓缩后得到的粗产品溶于DCM(10mL)和甲醇(3mL)的混合溶剂中,加入5-[(1R)-2-氨基-1-[叔丁基(二甲基)甲硅烷基]氧基甲基]-8-羟基-1H-喹啉-2-酮(0.2g,0.7mmol),室温搅拌30分钟后加入三乙酰氧基硼氢化钠(0.3g,1.4mmol),室温搅拌3小时。加入饱和碳酸氢钠水溶液(50mL),用DCM(100mL×2)萃取,合并后的有机相用无水硫酸钠干燥,减压浓缩后柱层析分离(DCM:甲醇(v/v)=1:0~8:1)得到黄色固体状的39G(0.3g,产率50%)。
LCMS m/z=428.4[M/2+1]。
第七步:N-[2-[[(2R)-2-羟基-2-(8-羟基-2-氧代-1H-喹啉-5-基)乙基]胺基]乙基]-3-[2-[3-[[4-(4-异丙基-2-吡啶基)-1-氧杂-4,9-杂螺二氮[5.5]十一烷-9-基]甲基]苯基]乙氧基]-N-甲基-丙酰胺(化合物39)
N-[2-[[(2R)-2-hydroxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]-3-[2-[3-[[4-(4-isopropyl-2-pyridyl)-1-oxa-4,9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]-N-methyl-propanamide
将39G(0.3g,0.4mmol)溶于THF(10mL)中,加入三乙胺三氢氟酸(0.6g,4.0mmol),室温搅拌12小时。依次滴加饱和碳酸氢钠水溶液(50mL),甲醇(5mL)淬灭反应,用DCM(100mL×2)萃取,合并后的有机相用无水硫酸钠干燥,减压浓缩后柱层析分离(DCM:甲醇(v/v)=1:0~8:1)得到黄色固体状的化合物39(0.1g,产率40%)。
1H NMR(400MHz,CD3OD)δ8.39-8.34(m,1H),7.96-7.95(d,1H),7.20-7.14(m,5H),6.98-6.97(d,1H),6.63-6.58(m,3H),5.20-5.17(m,1H),3.71-3.64(m,6H),3.48-3.35(m,6H),3.11(s,3H),2.84-2.75(m,8H),2.60-2.41(m,6H),1.92-1.89(m,2H),1.40-1.38(m,3H),1.23-1.21(m,6H).
LCMS m/z=742.5[M+1]。
实施例40:N-丁基-N-[2-[[(2R)-2-羟基-2-(8-羟基-2-氧代-1H-喹啉-5-基)乙基]氨基]乙基]-3-[2-[3-[2-[4-(2-吡啶基)-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-基]乙基]苯基]乙氧基]丙酰胺(化合物40)
N-butyl-N-[2-[[(2R)-2-hydroxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]-3-[2-[3-[2-[4-(2-pyridyl)-1-oxa-4,9-diazaspiro[5.5]undecan-9-yl]ethyl]phenyl]ethoxy]propanamide
第一步:3-[2-[3-[2-[4-(2-吡啶基)-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-基]乙基]苯基]乙氧基]丙酸叔丁酯(40B)
tert-butyl 3-[2-[3-[2-[4-(2-pyridyl)-1-oxa-4,9-diazaspiro[5.5]undecan-9-yl]ethyl]phenyl]ethoxy]propanoate
将3C(3.28g,14mmol)加入乙腈(50mL)中,加入3-[2-[3-(2-溴乙基)苯基]乙氧基]丙酸叔丁酯(中间体1)(4.9g,14mmol),碳酸钾(3.87g,28mmol),水(1mL),60℃搅拌24小时。待反应冷至室温后,加入水(50mL),用乙酸乙酯(100mL×2)萃取,合并有机相,有机相用无水硫酸钠干燥,过滤,滤液减压浓缩,残留物用硅胶柱层析纯化(DCM/甲醇(v/v)=40:1)得到黄色液体状的40B(4.28g,产率60%)。
第三步:N-丁基-N-(2,2-二甲氧基乙基)-3-[2-[3-[2-[4-(2-吡啶基)-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-基]乙基]苯基]乙氧基]丙酰胺(40C)
N-butyl-N-(2,2-dimethoxyethyl)-3-[2-[3-[2-[4-(2-pyridyl)-1-oxa-4,9-diazaspiro[5.5]undecan-9-yl]ethyl]phenyl]ethoxy]propanamide
将40B(2.2g,4.32mmol)溶于DCM(15mL)中,滴加三氟乙酸(10mL),室温搅拌4小时。减压浓缩除去反应溶剂,向残余物中加入甲苯(10mL),减压浓缩后,在0℃和氮气条件下向残留物中依次加入DCM(30mL)和三乙胺(1.98mL,14.2mmol),搅拌均匀后加入N-(2,2-二甲氧基乙基)丁-1-氨(0.84g,5.2mmol)(参考WO2011012896制备得到)和HATU(1.97g,5.18mmol),室温反应30分钟。加入水(30mL),DCM(30mL×2)萃取,合并有机相,饱和食盐水(40mL)洗,无水硫酸钠干燥,过滤,滤液减压浓缩,残留物经硅胶柱层析纯化(DCM/甲醇(v/v)=1:20)得到40C,透明液体(1.62g,收率63%)。
LCMS m/z=597.3[M+1]。
第四步:N-丁基-N-[2-[[(2R)-2-[叔丁基(二甲基)硅基]氧-2-(8-羟基-2-氧代-1H-喹啉-5-基)乙基]氨基]乙基]-3-[2-[3-[2-[4-(2-吡啶基)-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-基]乙基]苯基]乙氧基]丙酰胺(40D)
N-butyl-N-[2-[[(2R)-2-[tert-butyl(dimethyl)silyl]oxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]eth
yl]-3-[2-[3-[2-[4-(2-pyridyl)-1-oxa-4,9-diazaspiro[5.5]undecan-9-yl]ethyl]phenyl]ethoxy]propanamide
将40C(0.6g,1.0mmol)溶于THF(10mL)中,加入TsOH·H2O(0.96g,5.0mmol),40℃搅拌30min后,滴加饱和碳酸氢钠水溶液(50mL)淬灭反应,用DCM(50mL×2)萃取,合并有机相,无水硫酸钠干燥,过滤,滤液减压浓缩,向残留物中加入5-[2-氨基-1-(叔丁基(二甲基)硅基)氧基-乙基]-8-羟基-1H-喹啉-2-酮(0.34g,1.0mmol),甲醇(10mL)和DCM(10mL),室温搅拌1小时后,加入三乙酰氧基硼氢化钠(0.64g,3.0mmol),继续反应3小时。加入饱和碳酸氢钠溶液(30mL),DCM(30mL×2)萃取,合并有机相,无水硫酸钠干燥,过滤,滤液减压浓缩,残留物经硅胶柱层析纯化(DCM/甲醇(v/v)=1:12),得到黄色固体的40D(0.46g,产率53%).
LCMS m/z=435.4[M/2+1].
第五步:N-丁基-N-[2-[[(2R)-2-羟基-2-(8-羟基-2-氧代-1H-喹啉-5-基)乙基]氨基]乙基]-3-[2-[3-[2-[4-(2-吡啶基)-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-基]乙基]苯基]乙氧基]丙酰胺(化合物40)
N-butyl-N-[2-[[(2R)-2-hydroxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]-3-[2-[3-[2-[4-(2-pyridyl)-1-oxa-4,9-diazaspiro[5.5]undecan-9-yl]ethyl]phenyl]ethoxy]propanamide
将40D(0.46g,0.53mmol)溶于THF(10mL)中,加入三乙胺三氢氟酸盐(0.41mL,2.52mmol),室温反应16小时。反应结束后饱和碳酸氢钠溶液(20mL)淬灭反应,DCM(20mL×2)萃取,合并有机相,无水硫酸钠干燥,过滤,滤液减压浓缩,残留物经硅胶柱层析纯化(DCM/甲醇(v/v)=1:10),得到化合物40,黄色固体(0.21g,产率53%)。
1H NMR(400MHz,DMSO-d6)δ10.26(s,1H),8.18(d,1H),8.09(d,1H),7.52(s,1H),7.15(s,1H),7.05(s,3H),6.92(d,1H),6.83(d,1H),6.62(s,1H),6.48(d,1H),5.01(s,1H),3.74–3.66(m,2H),3.66–3.59(m,2H),3.56(m,2H),3.49–3.41(m,3H),3.36–3.23(m,6H),3.19(s,2H),2.79–2.59(m,6H),2.43–2.28(m,2H),2.00(s,1H),1.91(s,2H),1.73(s,2H),1.55(s,2H),1.46(s,1H),1.36(s,1H),1.32–1.16(m,8H).
LCMS m/z=378.3[M/2+1].
实施例41:3-[2-[2-氯-3-[[4-(4-乙基-2-吡啶基)-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-基]甲基]苯基]乙氧基]-N-[2-[[(2R)-2-羟基-2-8-羟基-2-氧代-1H-喹啉-5-基)乙基]氨基]乙基]-N-甲基-丙酰胺(化合物41)
3-[2-[2-chloro-3-[[4-(4-ethyl-2-pyridyl)-1-oxa-4,9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]-N-[2-[[(2R)-2-hydroxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]-N-methyl-propanamide
第一步:2-[2-氯-3-[[4-(4-乙基-2-吡啶基)-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-基]甲基]苯基]乙醇(41B)
2-[2-chloro-3-[[4-(4-ethyl-2-pyridyl)-1-oxa-4,9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethanol
取16D(1.3g,5mmol)溶于THF(30mL)中,加入2-氯-3-(2-羟基乙基)苯甲醛(参考CN201080043822制备得到)(0.92g,5mmol),室温搅拌1小时后加入三乙酰氧基硼氢化钠(3.2g,15mmol),室温搅拌3小时,加入饱和碳酸氢钠溶液(50mL)淬灭反应,DCM(60mL×2)萃取,合并有机相,有机相用无水硫酸钠干燥,过滤,滤液减压浓缩,残留物用硅胶柱层析纯化(DCM/甲醇(v/v)=40:1),得到棕色油状41B(1.27g,产率59%)。
第二步:3-[2-[2-氯-3-[[4-(4-乙基-2-吡啶基)-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-基]甲基]苯基]乙氧基]丙酸叔丁酯(41C)
tert-butyl 3-[2-[2-chloro-3-[[4-(4-ethyl-2-pyridyl)-1-oxa-4,9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]propanoate
取41B(1.27g,2.95mmol)溶于乙腈(1mL)中,加入丙烯酸叔丁酯(1.54g,12mmol)搅拌均匀后加入苄基三甲基氢氧化铵(0.17g,1mmol)(40%in MeOH),室温搅拌1小时。反应结束后体系减压浓缩,残留物用硅胶柱层析纯化(DCM/甲醇(v/v)=60:1),得到棕黄色油状的41C(1.1g,产率67%)。
第三步:3-[2-[2-氯-3-[[4-(4-乙基-2-吡啶基)-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-基]甲基]苯基]乙氧基]N-(2,2-二甲氧基乙基)-N-甲基-丙酰胺(41D)
3-[2-[2-chloro-3-[[4-(4-ethyl-2-pyridyl)-1-oxa-4,9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]-N-(2,2-dimethoxyethyl)-N-methyl-propanamide
取41C(0.56g,1mmol)加入DCM(20mL),0℃滴加三氟乙酸(8mL),滴毕,室温反应1小时后,反应液减压浓缩,加入甲苯(20mL)后减压浓缩。在0℃和氮气条件下向残留物中依次加入DCM(20mL)和三乙胺(0.4g,4mmol),搅拌均匀后加入甲氨基乙醛缩二甲醇(0.14g,1.2mmol)和HATU(0.46g,1.2mmol),室温反应30分钟。反应结束后加入水(30mL),DCM(30mL×2)萃取,合并有机相,饱和食盐水(40mL)洗,无水硫酸钠干燥,过滤,滤液减压浓缩,残留物经硅胶柱层析纯化(DCM/甲醇(v/v)=1:80)得到无色液体状的41D(0.53g,收率90%)。
第四步:N-[2-[[(2R)-2-[叔丁基(二甲基)硅基]氧基-2-(8-羟基-2-氧代-1H-喹啉-5-基)乙基]氨基]乙基]-3-[2-[2-氯-3-[[4-(4-乙基-2-吡啶基)-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-基]甲基]苯基]乙氧基]-N-甲基-丙酰胺(41E)
N-[2-[[(2R)-2-[tert-butyl(dimethyl)silyl]oxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]-3-[2-[2-chloro-3-[[4-(4-ethyl-2-pyridyl)-1-oxa-4,9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]-N-methyl-propanamide
取41D(0.53g,0.88mmol)溶于THF(20mL)中,加入TsOH·H2O(0.95g,5mmol),40℃反应30分钟。冷
却至室温,加入碳酸氢钠(0.67g,8mmol)的水溶液(10mL),搅拌10分钟后,加入饱和碳酸氢钠溶液(20mL),用乙酸乙酯(30mL×2)萃取,合并有机相,无水硫酸钠干燥,过滤,滤液减压浓缩,向残留物中加入5-[2-氨基-1-(叔丁基(二甲基)硅基)氧基-乙基]-8-羟基-1H-喹啉-2-酮(0.3g,0.9mmol),甲醇(10mL)和DCM(10mL),室温搅拌1小时后,加入三乙酰氧基硼氢化钠(0.64g,3mmol),继续反应3小时。加入饱和碳酸氢钠溶液(30mL)淬灭反应,用DCM(30mL×2)萃取,合并有机相,无水硫酸钠干燥,过滤,滤液减压浓缩,残留物经硅胶柱层析纯化(DCM/甲醇(v/v)=1:12),得到黄色固体状的41E(0.24g,产率31%)。
第五步:3-[2-[2-氯-3-[[4-(4-乙基-2-吡啶基)-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-基]甲基]苯基]乙氧基]-N-[2-[[(2R)-2-羟基-2-8-羟基-2-氧代-1H-喹啉-5-基)乙基]氨基]乙基]-N-甲基-丙酰胺(化合物41)
3-[2-[2-chloro-3-[[4-(4-ethyl-2-pyridyl)-1-oxa-4,9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]-N-[2-[[(2R)-2-hydroxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]-N-methyl-propanamide
取41E(0.24g,0.27mmol)溶于THF(20mL),加入三乙胺三氢氟酸盐(0.3mL),室温反应16小时。加入饱和碳酸氢钠溶液(20mL)淬灭反应,用DCM(20mL×2)萃取,合并有机相,无水硫酸钠干燥,过滤,滤液减压浓缩,残留物经硅胶柱层析纯化(DCM/甲醇(v/v)=1:10),得到淡黄色固体状的化合物41(0.09g,产率50%)。
LCMS m/z=381.3[M/2+1]。
1H NMR(400MHz,DMSO-d6)δ10.26(s,1H),8.17(dd,1H),7.97(d,1H),7.33(s,1H),7.31–7.15(m,2H),7.15–7.02(m,1H),6.92(dd,1H),6.67(s,1H),6.53–6.46(m,2H),5.30(s,1H),5.02(d,1H),3.73–3.67(m,2H),3.60(m,5H),3.47–3.42(m,2H),3.37(m,2H),2.94(s,3H),2.77(s,1H),2.72(m,4H),2.39(m,2H),2.05–1.91(m,1H),1.76(m,2H),1.57(s,2H),1.24(m,7H),1.15(m,3H),1.00(m,3H),0.85(s,1H).
实施例42:3-[2-[2-氯-3-[[4-(4-乙基-2-吡啶基)-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-基]甲基]苯基]乙氧基]-N-环戊基-N-[2-[[(2R)-2-羟基-2-8-羟基-2-氧代-1H-喹啉-5-基)乙基]氨基]乙基]-丙酰胺二三氟乙酸盐(化合物42)
3-[2-[2-chloro-3-[[4-(4-ethyl-2-pyridyl)-1-oxa-4,9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]-N-cyclopentyl-N-[2-[[(2R)-2-hydroxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]propanamide ditrifluoroacetic acid
第一步:3-[2-[2-氯-3-[[4-(4-乙基-2-吡啶基)-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-基]甲基]苯基]乙氧
基]N-环戊基-N-(2,2-二甲氧基乙基)丙酰胺(42A)
3-[2-[2-chloro-3-[[4-(4-ethyl-2-pyridyl)-1-oxa-4,9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]-N-cyclopentyl-N-(2,2-dimethoxyethyl)propanamide
取41C(0.56g,1mmol)加入DCM(20mL)中,0℃滴加三氟乙酸(8mL),滴毕,室温反应1小时后,反应液减压浓缩,加入甲苯(20mL)后减压浓缩。在0℃和氮气条件下向残留物中依次加入DCM(20mL)和三乙胺(0.4g,4mmol),搅拌均匀后加入环戊氨基乙醛缩二甲醇(0.25g,1.4mmol)和HATU(0.46g,1.2mmol),室温反应30分钟。加入水(30mL),用DCM(30mL×2)萃取,合并有机相,饱和食盐水(40mL)洗,无水硫酸钠干燥,过滤,滤液减压浓缩,残留物经硅胶柱层析纯化(DCM/甲醇(v/v)=1:80)得到透明液体状的42A(0.52g,收率80%)。
第二步:N-[2-[[(2R)-2-[叔丁基(二甲基)硅基]氧基-2-(8-羟基-2-氧代-1H-喹啉-5-基)乙基]氨基]乙基]-3-[2-[2-氯-3-[[4-(4-乙基-2-吡啶基)-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-基]甲基]苯基]乙氧基]-N-环戊基-丙酰胺(42B)
N-[2-[[(2R)-2-[tert-butyl(dimethyl)silyl]oxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]-3-[2-[2-chloro-3-[[4-(4-ethyl-2-pyridyl)-1-oxa-4,9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]-N-cyclopentyl-propanamide
取42A(0.52g,0.8mmol)溶于THF(20mL),加入TsOH·H2O(0.76g,4mmol),40℃反应30分钟。冷却至室温,加入饱和碳酸氢钠溶液(20mL),用乙酸乙酯(30mL×2)萃取,合并有机相,无水硫酸钠干燥,过滤,滤液减压浓缩,向残留物中加入5-[2-氨基-1-(叔丁基(二甲基)硅基)氧基-乙基]-8-羟基-1H-喹啉-2-酮(0.27g,0.8mmol),甲醇(10mL)和DCM(10mL),室温搅拌1小时后,加入三乙酰氧基硼氢化钠(0.64g,3mmol),继续反应3小时。反应结束后饱和碳酸氢钠溶液(30mL)淬灭反应,用DCM(30mL×2)萃取,合并有机相,无水硫酸钠干燥,过滤,滤液减压浓缩,残留物经硅胶柱层析纯化(DCM/甲醇(v/v)=1:12),得到黄色固体状的42B(0.23g,产率31%)。
第三步:3-[2-[2-氯-3-[[4-(4-乙基-2-吡啶基)-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-基]甲基]苯基]乙氧基]-N-环戊基-N-[2-[[(2R)-2-羟基-2-8-羟基-2-氧代-1H-喹啉-5-基)乙基]氨基]乙基]-丙酰胺二三氟乙酸盐(化合物42)
3-[2-[2-chloro-3-[[4-(4-ethyl-2-pyridyl)-1-oxa-4,9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]-N-cyclopentyl-N-[2-[[(2R)-2-hydroxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]propanamide ditrifluoroacetic acid
取42B(0.23g,0.25mmol)溶于THF(20mL),加入三乙胺三氢氟酸盐(0.3mL),室温反应16小时。加入饱和碳酸氢钠溶液(20mL)淬灭反应,用DCM(20mL×2)萃取,合并有机相,无水硫酸钠干燥,过滤,滤液减压浓缩,残留物经硅胶柱层析纯化(DCM/甲醇(v/v)=1:10)后用液相制备柱分离提纯(液相制备条件:C18反相制备柱,流动相为含0.1%三氟乙酸的去离子水(A),含0.1%三氟乙酸的乙腈(B),梯度洗脱,B含量5%~50%,洗脱时间15min,流速12mL/min,柱温:30℃)得到白色固体状的化合物42(0.04g,产率20%)。
1H NMR(400MHz,DMSO-d6)δ10.48(s,2H),9.61(s,1H),8.63(s,2H),8.18(d,1H),7.98(d,1H),7.36
(s,3H),7.14(d,1H),6.98(d,1H),6.91(s,1H),6.70(s,1H),6.58(d,1H),6.16(d,2H),5.39–5.24(m,2H),4.41(s,3H),4.30(s,3H),4.17(s,3H),3.77(s,2H),3.63(s,3H),3.52(s,2H),3.46(s,3H),3.23(s,2H),3.06(s,4H),2.82(s,2H),2.59(s,3H),2.11(s,2H),2.00(s,1H),1.79(s,1H),1.78–1.61(m,3H),1.49(s,3H),1.24(s,3H).
LCMS m/z=815.3[M+1]。
实施例43:3-[2-[3-[[4-(4-乙基-2-吡啶基)-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-基]甲基]苯基]乙氧基]-N-[2-[[(2R)-2-羟基-2-(5-羟基-3-氧-4H-1,4-苯并噁嗪-8-基)乙基]氨基]乙基]-N-甲基-丙酰胺二三氟乙酸盐(化合物43)
3-[2-[3-[[4-(4-ethyl-2-pyridyl)-1-oxa-4,9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]-N-[2-[[(2R)-2-hydroxy-2-(5-hydroxy-3-oxo-4H-1,4-benzoxazin-8-yl)ethyl]amino]ethyl]-N-methyl-propanamide ditrifluoroacetic acid
第一步:N-[2-[[(2R)-2-[叔丁基(二甲基)硅基]氧基-2-(5-羟基-3-氧-4H-1,4-苯并噁嗪-8-基)乙基]氨基]乙基]-3-[2-[3-[[4-(4-乙基-2-吡啶基)-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-基]甲基]苯基]乙氧基]-N-甲基-丙酰胺(43A)
N-[2-[[(2R)-2-[tert-butyl(dimethyl)silyl]oxy-2-(5-hydroxy-3-oxo-4H-1,4-benzoxazin-8-yl)ethyl]amino]ethyl]-3-[2-[3-[[4-(4-ethyl-2-pyridyl)-1-oxa-4,9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]-N-methyl-propanamide
取18C(0.52g,0.91mmol)溶于THF(20mL),加入TsOH·H2O(0.95g,5mmol),40℃反应30分钟,冷却至室温,加入饱和碳酸氢钠溶液(20mL),用乙酸乙酯(30mL×2)萃取,合并有机相,无水硫酸钠干燥,过滤,滤液减压浓缩,向残留物中加入8-[(1R)-2-氨基-1-[叔丁基(二甲基)硅基]氧基-乙基]-5-羟基-4H-1,4-苯并噁嗪-3-酮(0.3g,0.9mmol),甲醇(10mL)和DCM(10mL),室温搅拌1小时后,加入三乙酰氧基硼氢化钠(0.64g,3mmol),继续反应3小时。反应结束后饱和碳酸氢钠溶液(30mL)淬灭反应,用DCM(30mL×2)萃取,合并有机相,无水硫酸钠干燥,过滤,滤液减压浓缩,残留物经硅胶柱层析纯化(DCM/甲醇(v/v)=1:12),得到白色固体状的43A(0.32g,产率41%)。
LCMS m/z=845.5[M+1].
第二步:3-[2-[3-[[4-(4-乙基-2-吡啶基)-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-基]甲基]苯基]乙氧基]-N-[2-[[(2R)-2-羟基-2-(5-羟基-3-氧-4H-1,4-苯并噁嗪-8-基)乙基]氨基]乙基]-N-甲基-丙酰胺二三氟乙酸盐(化合物43)
3-[2-[3-[[4-(4-ethyl-2-pyridyl)-1-oxa-4,9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]-N-[2-[[(2R)-2-hydroxy-2-(5-hydroxy-3-oxo-4H-1,4-benzoxazin-8-yl)ethyl]amino]ethyl]-N-methyl-propanamide
ditrifluoroacetic acid
取43A(0.32g,0.38mmol)溶于THF(20mL),加入三乙胺三氢氟酸盐(0.4mL),室温反应16小时,加入饱和碳酸氢钠溶液(20mL)淬灭反应,用DCM(20mL×2)萃取,合并有机相,无水硫酸钠干燥,过滤,滤液减压浓缩,残留物经硅胶柱层析纯化(DCM/甲醇(v/v)=1:10)后用液相制备柱分离提纯(液相制备条件:C18反相制备柱,流动相为含0.1%三氟乙酸的去离子水(A),含0.1%三氟乙酸的乙腈(B),梯度洗脱,B含量=5%~50%,洗脱时间15min,流速12mL/min,柱温:30℃)得到白色固体状的化合物43(90mg,产率20%)。
1H NMR(400MHz,DMSO)δ9.96(s,2H),9.54(s,1H),8.57(s,1H),8.44(s,1H),8.00(d,1H),7.37(s,3H),6.97–6.79(m,2H),6.66(s,1H),6.56(s,1H),5.92(s,1H),5.33(s,1H),5.04(d,1H),4.53(s,2H),4.45(s,2H),4.31(s,2H),3.76(s,2H),3.69–3.54(m,5H),3.50(s,2H),3.44(s,2H),3.23(s,2H),3.07(s,3H),2.95(s,3H),2.81(s,2H),2.56(s,3H),2.11(d,2H),2.00(s,1H),1.77–1.62(m,2H),1.45(s,1H),1.24(s,4H).
LCMS m/z=366.3[M/2+1]。
实施例44:3-[2-[3-[[4-(4-乙基-2-吡啶基)-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-基]甲基]苯基]乙氧基]-N-[2-[2-(5-羟基-3-氧-4H-1,4-苯并噁嗪-8-基)乙基氨基]乙基]-N-甲基-丙酰胺二三氟乙酸盐(化合物44)
3-[2-[3-[[4-(4-ethyl-2-pyridyl)-1-oxa-4,9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]-N-[2-[2-(5-hydroxy-3-oxo-4H-1,4-benzoxazin-8-yl)ethylamino]ethyl]-N-methyl-propanamide ditrifluoroaceticacid
取18C(0.52g,0.91mmol)溶于THF(20mL)中,加入TsOH·H2O(0.95g,5mmol),40℃反应30分钟,冷却至室温,加入饱和碳酸氢钠溶液(20mL),乙酸乙酯(30mL×2)萃取,合并有机相,无水硫酸钠干燥,过滤,滤液减压浓缩,向残留物中加入8-(2-氨乙基)-5-羟基-4H-1,4-苯并噁嗪-3-酮的乙酸盐(0.27g,1mmol),甲醇(10mL),N-甲基吡咯烷酮(10mL)和乙酸(0.06g,1mmol),室温搅拌1小时后,加入三乙酰氧基硼氢化钠(0.64g,3mmol),反应3小时。加入饱和碳酸氢钠溶液(30mL),用DCM(30mL×2)萃取,合并有机相,无水硫酸钠干燥,过滤,滤液减压浓缩残留物经硅胶柱层析纯化(DCM/甲醇(v/v)=1:14)后用液相制备柱分离提纯(液相制备条件:C18反相制备柱,流动相为含0.1%三氟乙酸的去离子水(A),含0.1%三氟乙酸的乙腈(B),梯度洗脱,B含量=5%~50%,洗脱时间15min,流速12mL/min,柱温:30℃)得到白色固体状的化合物44(0.12g,产率12%)。
1H NMR(400MHz,DMSO-d6)δ9.93(s,1H),9.85(s,1H),9.53(s,1H),8.53(s,1H),8.37(s,1H),8.00(d,1H),7.36(m,3H),6.86(s,1H),6.74–6.57(m,2H),6.48(m,1H),5.37–5.28(m,1H),4.54(s,2H),4.40(s,1H),4.31(s,2H),3.77(s,2H),3.68–3.58(m,3H),3.52(m,3H),3.44(s,1H),3.23(d,2H),3.06(s,5H),2.95(s,2H),2.82(m,3H),2.56(m,2H),2.12(d,2H),2.00(m,2H),1.84(s,1H),1.75–1.62(m,2H),1.47(m,1H),1.24(s,4H).
LCMS m/z=715.4[M+1]。
实施例45:N-(环丙甲基)-3-[2-[3-[[4-(4-乙基-2-吡啶基)-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-基]甲基]苯基]乙氧基]-N-[2-[[(2R)-2-羟基-2-(8-羟基-2-氧代-1H-喹啉-5-基)乙基]氨基]乙基]丙酰胺二三氟乙酸盐(化合物45)
N-(cyclopropylmethyl)-3-[2-[3-[[4-(4-ethyl-2-pyridyl)-1-oxa-4,9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]-N-[2-[[(2R)-2-hydroxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]propanamide ditrifluoroacetic acid
第一步:N-(环丙甲基)-N-(2,2-二甲氧基乙基)-3-[2-[3-[[4-(4-乙基-2-吡啶基)-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-基]甲基]苯基]乙氧基]丙酰胺(45A)
N-(cyclopropylmethyl)-N-(2,2-dimethoxyethyl)-3-[2-[3-[[4-(4-ethyl-2-pyridyl)-1-oxa-4,9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]propanamide
取18A(0.52g,1mmol)加入DCM(20mL),0℃滴加三氟乙酸(8mL),滴毕,室温反应1小时后,反应液减压浓缩,加入甲苯(20mL)后减压浓缩。在0℃和氮气条件下向残留物中依次加入DCM(20mL)和三乙胺(0.4g,4mmol),搅拌均匀后加入环丙甲氨基乙醛缩二甲醇(0.18g,1.1mmol)和HATU(0.46g,1.2mmol),室温反应30分钟。加入水(30mL),用DCM(30mL×2)萃取,合并有机相,饱和食盐水(40mL)洗,无水硫酸钠干燥,过滤,滤液减压浓缩,残留物经硅胶柱层析纯化(DCM/甲醇(v/v)=1:60)得到无色液体状的45A(0.51g,收率83%)。
LCMS m/z=609.4[M+1]。
第二步:N-[2-[[(2R)-2-[叔丁基(二甲基)硅基]氧基-2-(8-羟基-2-氧代-1H-喹啉-5-基)乙基]氨基]乙基]-N-(环丙甲基)-3-[2-[3-[[4-(4-乙基-2-吡啶基)-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-基]甲基]苯基]乙氧基]丙酰胺(45B)
N-[2-[[(2R)-2-[tert-butyl(dimethyl)silyl]oxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]-N-(cyclopropylmethyl)-3-[2-[3-[[4-(4-ethyl-2-pyridyl)-1-oxa-4,9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]propanamide
将45A(0.51g,0.84mmol)溶于THF(20mL),加入TsOH·H2O(0.8g,4.2mmol),40℃反应30分钟,冷却至室温,加入饱和碳酸氢钠溶液(20mL),乙酸乙酯(30mL×2)萃取,合并有机相,无水硫酸钠干燥,
过滤,滤液减压浓缩,向残留物中加入5-[2-氨基-1-(叔丁基(二甲基)硅基)氧基-乙基]-8-羟基-1H-喹啉-2-酮(0.28g,0.84mmol),甲醇(10mL)和DCM(10mL),室温搅拌1小时后,加入三乙酰氧基硼氢化钠(0.64g,3mmol),继续反应3小时。反应结束后饱和碳酸氢钠溶液(30mL)淬灭反应,DCM(30mL×2)萃取,合并有机相,无水硫酸钠干燥,过滤,滤液减压浓缩,残留物经硅胶柱层析纯化(DCM/甲醇(v/v)=1:12),得到黄色固体状的45B(0.29g,产率39%)。
LCMS m/z=441.4[M/2+1]
第三步:N-(环丙甲基)-3-[2-[3-[[4-(4-乙基-2-哌啶基)-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-基]甲基]苯基]乙氧基]-N-[2-[[(2R)-2-羟基-2-(8-羟基-2-氧代-1H-喹啉-5-基)乙基]氨基]乙基]丙酰胺二三氟乙酸盐(化合物45)
N-(cyclopropylmethyl)-3-[2-[3-[[4-(4-ethyl-2-pyridyl)-1-oxa-4,9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]-N-[2-[[(2R)-2-hydroxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]propanamide ditrifluoroacetic acid
取45B(0.29g,0.33mmol)溶于THF(20mL),加入三乙胺三氢氟酸盐(0.3mL),室温反应16小时。加入饱和碳酸氢钠溶液(20mL),DCM(20mL×2)萃取,合并有机相,无水硫酸钠干燥,过滤,滤液减压浓缩,残留物经硅胶柱层析纯化(DCM/甲醇(v/v)=1:10)后用液相制备柱分离提纯(液相制备条件:C18反相制备柱,流动相为含0.1%三氟乙酸的去离子水(A),含0.1%三氟乙酸的乙腈(B),梯度洗脱,B含量=5%~50%,洗脱时间15min,流速12mL/min,柱温:30℃)得到白色固体状的化合物45(0.06g,产率20%)。
1H NMR(400MHz,DMSO-d6)δ10.49(s,2H),9.55(s,1H),8.71(s,1H),8.57(s,1H),8.16(m,1H),7.99(m,1H),7.36(s,3H),7.15(m,1H),7.03–6.93(m,1H),6.85(s,1H),6.60(m,2H),6.17(s,2H),5.43–5.22(m,2H),4.30(s,3H),3.76(s,4H),3.64(m,6H),3.44(s,3H),3.14(m,7H),2.82(m,2H),2.71–2.55(m,3H),2.11(m,2H),2.00(m,2H),1.83(s,1H),1.69(m,2H),1.40(s,1H),1.24(s,5H).
LCMS m/z=384.4[M/2+1].
实施例46:N-环戊基-3-[2-[3-[[4-(4-乙基-2-吡啶基)-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-基]甲基]苯基]乙氧基]-N-[2-[[(2R)-2-羟基-2-(8-羟基-2-氧代-1H-喹啉-5-基)乙基]氨基]乙基]丙酰胺二三氟乙酸盐(化合物46)
N-cyclopentyl-3-[2-[3-[[4-(4-ethyl-2-pyridyl)-1-oxa-4,9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]-N-[2-[[(2R)-2-hydroxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]propanamide ditrifluoroaceticacid
第一步:N-环戊基-N-(2,2-二甲氧基乙基)-3-[2-[3-[[4-(4-乙基-2-吡啶基)-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-基]甲基]苯基]乙氧基]丙酰胺(46A)
N-cyclopentyl-N-(2,2-dimethoxyethyl)-3-[2-[3-[[4-(4-ethyl-2-pyridyl)-1-oxa-4,9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]propanamide
取18A(0.52g,1mmol)加入DCM(20mL),0℃滴加三氟乙酸(8mL),滴毕,室温反应1小时后,将体系减压浓缩,加入甲苯(20mL)后减压浓缩。在0℃和氮气条件下向残留物中依次加入DCM(20mL)和三乙胺(0.4g,4mmol),搅拌均匀后加入环戊氨基乙醛缩二甲醇(0.19g,1.1mmol)和HATU(0.46g,1.2mmol),室温反应30分钟。加入水(30mL),DCM(30mL×2)萃取,合并有机相,饱和食盐水(40mL)洗涤,无水硫酸钠干燥,过滤,滤液减压浓缩,残留物经硅胶柱层析纯化(DCM/甲醇(v/v)=1:60)得到无色液体状的46A(0.56g,收率89%)。
LCMS m/z=623.4[M+1]
第二步:N-[2-[[(2R)-2-[叔丁基(二甲基)硅基]氧基-2-(8-羟基-2-氧代-1H-喹啉-5-基)乙基]氨基]乙基]-N-环戊基-3-[2-[3-[[4-(4-乙基-2-吡啶基)-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-基]甲基]苯基]乙氧基]丙酰胺(46B)
N-[2-[[(2R)-2-[tert-butyl(dimethyl)silyl]oxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]-N-cyclopentyl-3-[2-[3-[[4-(4-ethyl-2-pyridyl)-1-oxa-4,9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]propanamide
取46A(0.56g,0.9mmol)溶于THF(20mL),加入TsOH·H2O(0.95g,5mmol),40℃反应30分钟,冷却至室温,加入饱和碳酸氢钠溶液(30mL),乙酸乙酯(30mL×2)萃取,合并有机相,无水硫酸钠干燥,过滤,滤液减压浓缩,向残留物中加入5-[2-氨基-1-(叔丁基(二甲基)硅基)氧基-乙基]-8-羟基-1H-喹啉-2-酮(0.3g,0.9mmol),甲醇(10mL)和DCM(10mL),室温搅拌1小时后,加入三乙酰氧基硼氢化钠(0.64g,3mmol),继续反应3小时。反应结束后饱和碳酸氢钠溶液(30mL)淬灭反应,DCM(30mL×2)萃取,合并有机相,无水硫酸钠干燥,过滤,滤液减压浓缩,残留物经硅胶柱层析纯化(DCM/甲醇(v/v)=1:12),得到46B,黄色固体(0.35g,产率:43%)。
LCMS m/z=448.4[M/2+1]。
第三步:N-环戊基-3-[2-[3-[[4-(4-乙基-2-吡啶基)-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-基]甲基]苯基]乙氧基]-N-[2-[[(2R)-2-羟基-2-(8-羟基-2-氧代-1H-喹啉-5-基)乙基]氨基]乙基]丙酰胺二三氟乙酸盐(化合物46)
N-cyclopentyl-3-[2-[3-[[4-(4-ethyl-2-pyridyl)-1-oxa-4,9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]-N-[2-[[(2R)-2-hydroxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]propanamide ditrifluoroaceticacid
将46B(0.35g,0.39mmol)溶于THF(20mL),加入三乙胺三氢氟酸盐(0.4mL),室温反应16小时。加入饱和碳酸氢钠溶液(20mL),DCM(20mL×2)萃取,合并有机相,无水硫酸钠干燥,过滤,滤液减压浓缩,残留物经硅胶柱层析纯化(DCM/甲醇(v/v)=1:10)后用液相制备柱分离提纯(液相制备条件:C18反相制备柱,流动相为含0.1%三氟乙酸的去离子水(A),含0.1%三氟乙酸的乙腈(B),梯度洗脱,B含量=5%~50%,洗脱时间15min,流速12mL/min,柱温:30℃)得到白色固体状的化合物46(0.11g,产率:28%)。
1H NMR(400MHz,DMSO-d6)δ10.48(s,2H),9.61(s,1H),8.63(s,2H),8.18(d,1H),7.98(d,1H),7.36(s,3H),7.14(d,1H),6.98(d,1H),6.91(s,1H),6.70(s,1H),6.58(d,1H),6.29–6.04(m,1H),5.39–5.22(m,2H),4.41(s,1H),4.30(s,2H),4.17(s,1H),3.77(s,2H),3.63(d,4H),3.52(s,2H),3.46(s,3H),3.35–2.94(m,7H),2.82(s,2H),2.59(s,3H),2.33(s,1H),2.11(d,2H),2.02–1.89(m,1H),1.79(s,1H),1.74–1.60(m,3H),1.49(s,4H),1.24(s,3H).
LCMS m/z=391.3[M/2+1]。
实施例47:N-[2-[[(2R)-2-羟基-2-(8-羟基-2-氧代-1H-喹啉-5-基)乙基]氨基]乙基]-N-甲基-3-[2-[3-[2-[4-(2-氧代-1H-吡啶基-3-基)-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-基]乙基]苯基]乙氧基]丙酰胺(化合物47)
N-[2-[[(2R)-2-hydroxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]-N-methyl-3-[2-[3-[2-[4-(2-oxo-1H-pyridin-3-yl)-1-oxa-4,9-diazaspiro[5.5]undecan-9-yl]ethyl]phenyl]ethoxy]propanamide
第一步:4-(2-苄氧-3-吡啶基)-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-羧酸叔丁酯(47B)
tert-butyl 4-(2-benzyloxy-3-pyridyl)-1-oxa-4,9-diazaspiro[5.5]undecane-9-carboxylate
将1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-羧酸叔丁酯(0.512g,2.00mmol)加入甲苯(50mL)中,加入2-苄氧-3-溴吡啶(47A)(0.528g,2.00mmol),叔丁醇钠(0.40g,4.16mmol),4,5-双二苯基膦-9,9-二甲基氧
杂蒽(0.036g,0.062mmol),三(二亚苄基丙酮)二钯(0.036g,0.039mmol),氮气氛100℃反应6小时。反应液冷却至室温,加入乙酸乙酯(200mL)和水(100mL),萃取,有机相和水相分离,水相用乙酸乙酯(100mL)萃取,合并后的有机相用饱和食盐水(100mL)洗涤,无水硫酸钠干燥,减压浓缩后柱层析分离(石油醚/乙酸乙酯(v/v)=1:1)得到黄色液体状的47B(0.812g,产率93%)。
LCMS m/z=440.2[M+1]。
第二步:4-(2-氧代-1H-吡啶-3-基)-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-羧酸叔丁酯(47C)
tert-butyl 4-(2-oxo-1H-pyridin-3-yl)-1-oxa-4,9-diazaspiro[5.5]undecane-9-carboxylate
将47B(0.89g,2.02mmol)加入乙酸乙酯(20mL)中,加入钯碳(0.08g,10%w/w),氢气氛反应4小时。反应液用硅藻土过滤,用乙酸乙酯(100ml)洗涤滤饼,滤液减压浓缩后柱层析分离(石油醚/乙酸乙酯(v/v)=1:1)得到黄色液体状的47C(0.700g,产率99%)。
LCMS m/z=372.2[M+23]。
第三步:3-(1-氧杂-4,9-二氮杂螺[5.5]十一烷-4-基)-1H-吡啶-2-酮(47D)
3-(1-oxa-4,9-diazaspiro[5.5]undecan-4-yl)-1H-pyridin-2-one
将47C(0.70g,2.00mmol)溶于DCM(20mL)中,加入三氟乙酸(4mL),室温搅拌2小时。减压浓缩除去反应溶剂,向残余物中加入甲苯(5mL),减压浓缩后得到黄色液体状的47D的三氟乙酸盐(0.50g,产率100%)。
LCMS m/z=250.2[M+1].
第四步:3-[2-[3-[2-[4-(2-氧-1H-吡啶-3-基)-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-基]乙基]苯基]乙氧基]丙酸叔丁酯(47E)
tert-butyl 3-[2-[3-[2-[4-(2-oxo-1H-pyridin-3-yl)-1-oxa-4,9-diazaspiro[5.5]undecan-9-yl]ethyl]phenyl]ethoxy]propanoate
将47D的三氟乙酸盐(0.500g,2.00mmol)置于乙腈(25mL)中,加入中间体1(0.720g,2.00mmol),碳酸钾(1.589g,11.50mmol),水(0.5mL)。60℃搅拌24小时。待反应冷至室温后,加入水(50mL),用乙酸乙酯(100mL×2)萃取,合并后的有机相用无水硫酸钠干燥,减压浓缩后柱层析分离(DCM/甲醇(v/v)=15:1)得到黄色液体状的47E(0.43g,产率41%)。
LCMS m/z=526.3[M+1]。
第五步:3-[2-[3-[2-[4-(2-氧代-1-H-吡啶-3-基)-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-基]乙基]苯基]乙氧基]丙酸(47F)
3-[2-[3-[2-[4-(2-oxo-1H-pyridin-3-yl)-1-oxa-4,9-diazaspiro[5.5]undecan-9-yl]ethyl]phenyl]ethoxy]propanoic acid
将47E(0.43,0.818mmol)加入DCM(15mL)中,滴加三氟乙酸(5mL),室温搅拌4小时。减压浓缩除去反应溶剂,向残余物中加入甲苯(10mL),减压浓缩后得到黄色液体状的47F的三氟乙酸盐(0.38,产率99%)。
LCMS m/z=470.3[M+1]。
第六步:N-(2,2-二甲氧基乙基)-N-甲基-3-[2-[3-[2-[4-(2-氧代-1H-吡啶-3-基)-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-基]乙基]苯基]乙氧基]丙酰胺(47G)
N-(2,2-dimethoxyethyl)-N-methyl-3-[2-[3-[2-[4-(2-oxo-1H-pyridin-3-yl)-1-oxa-4,9-diazaspiro[5.5]undecan-9-yl]ethyl]phenyl]ethoxy]propanamide
将47F的三氟乙酸盐(0.38g,0.81mmol)溶于DCM(20mL)中,加入三乙胺(0.246g,2.43mmol),甲氨基乙醛缩二甲醇(0.116g,0.973mmol),HATU(0.462g,1.21mmol),室温搅拌2小时。滴加饱和碳酸氢钠水溶液(50mL),用DCM(100mL×2)萃取,合并后的有机相用饱和食盐水(50mL)洗涤,无水硫酸钠干燥,减压浓缩后柱层析分离(DCM/甲醇(v/v)=15:1)得到黄色液体状的47G(0.46g,产率99%)。
LCMS m/z=571.4[M+1]。
第七步:N-[2-[[(2R)-2-(叔丁基(二甲基)硅基)氧基-2-(8-羟基-2-氧代-1H-喹啉-5-基)乙基]氨基]乙基]-N-甲基-3-[2-[3-[2-[4-(2-氧代-1H-吡啶-3-基)-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-基]乙基]苯基]乙氧基]丙酰胺(47H)
N-[2-[[(2R)-2-(tert-butyl(dimethyl)silyl)oxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]-N-methyl-3-[2-[3-[2-[4-(2-oxo-1H-pyridin-3-yl)-1-oxa-4,9-diazaspiro[5.5]undecan-9-yl]ethyl]phenyl]ethoxy]propanamide
将47G(0.465g,0.81mmol)溶于THF(10mL)中,加入TsOH·H2O(0.77g,4.04mmol),40℃搅拌1小时后,滴加饱和碳酸氢钠水溶液(50mL)淬灭反应,用DCM(100mL×2)萃取,合并后的有机相用无水硫酸钠干燥,减压浓缩后得到的粗产品溶于DCM(10mL)和甲醇(3mL)的混合溶剂中,加入5-[2-氨基-1-(叔丁基(二甲基)硅基)氧基-乙基]-8-羟基-1H-喹啉-2-酮(0.27g,0.80mmol),室温搅拌30分钟后加入三乙酰氧基硼氢化钠(0.51g,2.42mmol),室温搅拌3小时。加入饱和碳酸氢钠水溶液(50mL)淬灭反应,用DCM(100mL×2)萃取,合并后的有机相用无水硫酸钠干燥,减压浓缩后柱层析分离(DCM:甲醇(v/v)=1:0~8:1)得到黄色固体状的47H(0.40g,产率59%)。
LCMS m/z=422.4[M/2+1]。
第八步:N-[2-[[(2R)-2-羟基-2-(8-羟基-2-氧代-1H-喹啉-5-基)乙基]氨基]乙基]-N-甲基-3-[2-[3-[2-[4-(2-氧代-1H-吡啶基-3-基)-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-基]乙基]苯基]乙氧基]丙酰胺(化合物47)
N-[2-[[(2R)-2-hydroxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]-N-methyl-3-[2-[3-[2-[4-(2-oxo-1H-pyridin-3-yl)-1-oxa-4,9-diazaspiro[5.5]undecan-9-yl]ethyl]phenyl]ethoxy]propanamide
将47H(0.40g,0.474mmol)溶于THF(10mL)中,加入三乙胺三氢氟酸(0.22g,1.1mmol),室温搅拌12小时。依次滴加饱和碳酸氢钠水溶液(50mL),甲醇(5mL)淬灭反应,用DCM(100mL×2)萃取,合并后的有机相用无水硫酸钠干燥,减压浓缩后柱层析分离(DCM:甲醇(v/v)=1:0~8:1)得到黄色固体状的化合物47(0.076g,产率22%)。
1H NMR(400MHz,CD3OD)δ8.37(dd,1H),7.28–7.14(m,2H),7.13–6.96(m,5H),6.91(d,1H),6.63(t,1H),6.34(t,1H),5.24(dt,1H),3.93–3.79(m,2H),3.73(dd,2H),3.69–3.42(m,5H),3.13–2.74(m,20H),2.63(t,2H),2.20(d,2H),1.85(t,2H).
LCMS m/z=730.4[M+1].
实施例48:本发明化合物48-化合物138,化合物(可用通式I概括)的合成方法如下:
步骤1):化合物A(LG=Cl、Br、I或其它离去基团)(1.5eq.)与1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-羧酸叔丁酯(1.0eq.)加入溶剂中(如1,4-二氧六环,DMF,乙腈),在条件1(叔丁醇钠(2.0eq.),4,5-双二苯基膦-9,9-二甲基氧杂蒽(0.15eq.)和醋酸钯(0.07eq.),氮气氛下回流反应)或条件2(碳酸钾(2.0eq.)或三乙胺(3.0eq.),40℃~反应回流)下反应,可根据反应监控情况,增加反应温度)下反应,反应至TLC或LC-MS监控反应结束。采用常规后处理如萃取、柱层析分离得到化合物B;
步骤2):化合物B溶于DCM中,加入三氟乙酸(DCM体积的1/4),室温反应至TLC或LC-MS监控反应结束。直接减压浓缩得到化合物C的三氟乙酸盐(直接用于下一步反应),或者采用常规后处理如加入水,调节溶液pH值至中性,萃取、柱层析分离得到化合物C。
步骤3)化合物C或者其三氟乙酸盐(1.0eq)溶于乙腈中,加入1.0eq中间体1~6中的任一个、5.0eq碳酸钾和0.5mL水,40℃~60℃反应至TLC或LC-MS监控反应结束。用常规后处理如萃取、柱层析分离得到化合物D。
步骤4)化合物D溶于DCM中,加入三氟乙酸(DCM体积的1/3),室温反应至TLC或LC-MS监控反应结束。直接减压浓缩得到化合物E(直接用于下一步反应),或者采用常规后处理如加入水,调节溶液pH值至中性,萃取、柱层析分离得到化合物E。
步骤5)化合物E(1.0eq)溶于DCM中,加入化合物F(Rx=甲基、乙基或环戊基)(1.5eq)、三乙胺(3~5eq.)和HATU(1.5eq.),室温反应至TLC或LC-MS监控反应结束。加入饱和碳酸氢钠水溶液淬灭反应,萃取、柱层析分离得到化合物G。
步骤6)化合物G(1.0eq.)溶于THF中,向其中加入TsOH·H2O(5.0eq),40℃反应至TLC或LC-MS
监控反应结束。加入饱和碳酸氢钠水溶液淬灭反应,萃取、减压浓缩得到的产物溶于DCM和甲醇的混合溶剂(v/v=10/3),加入化合物H(1.0eq),室温搅拌30min后,加入三乙酰氧基硼氢化钠(3.0eq.),室温反应至TLC或LC-MS监控反应结束。加入饱和碳酸氢钠水溶液淬灭反应,萃取、柱层析分离得到产品。当化合物H中R9=H时,此时得到的产品即为化合物I。当化合物H中R9=OTBS时,还需将产品(1.0eq.)进一步溶于THF中,加入三乙胺三氢氟酸(1.1eq.),室温反应至TLC或LC-MS监控反应结束。加入饱和碳酸氢钠水溶液淬灭反应,萃取、柱层析分离得到化合物I。
化合物I合成中,萃取后浓缩后的残余物也可用制备液相色谱分离提纯,色谱条件:C18反相制备柱,流动相为含0.1%三氟乙酸的去离子水(A),含0.1%三氟乙酸的乙腈(B),梯度洗脱B:(A+B)=5%~20%,洗脱时间15min,流速12mL/min,柱温:30℃),通过制备液相色谱分离提纯后得到的产品化合物I为二三氟乙酸盐形式。本领域技术人员可根据常规手段更改色谱条件,分离纯化产品。
同时,化合物48-化合物149(可用通式I概括)也可使用实施例1~47和中间体1~15中的方法合成所需要的中间体。
化合物48
1H NMR(400MHz,DMSO-d6)δ10.23(br,1H),8.17(d,1H),8.05(t,2H),7.56(t,1H),7.45(t,1H),7.19-7.12(m,1H),7.10-6.98(m,4H),6.92(d,1H),6.48(dd,1H),5.05-4.96(m,1H),3.89–3.78(m,2H),3.64-3.52(m,4H),3.40-3.36(m,2H),3.34-3.29(m,2H),3.26(s,2H),2.92(s,1H),2.78–2.64(m,9H),2.62-2.52(m,6H),2.36(t,2H),1.95(d,2H),1.66(t,2H),1.23(s,2H).
LCMS m/z=385.3[M/2+1].
化合物49
1H NMR(400MHz,DMSO-d6)δ10.28(br,1H),8.17(dd,1H),7.82(d,1H),7.18(s,1H),7.14–7.04(m,4H),6.94–6.88(m,1H),6.55(s,1H),6.48(dd,1H),5.05-4.97(m,1H),4.04-3.99(m,2H),3.96-3.87(m,2H),3.66–3.52(m,8H),3.38-3.24(m,14H),2.80–2.61(m,6H),2.38-2.24(m,4H),1.71-1.63(m,2H),1.58–1.40(m,4H),0.56-0.49(m,2H),0.40-0.33(m,2H).
LCMS m/z=406.8[M/2+1].
化合物50
1H NMR(400MHz,DMSO-d6)δ10.26(br,1H),8.18(d,1H),7.94(d,1H),7.22–7.04(m,6H),6.91(d,1H),6.65(s,1H),6.50–6.45(m,2H),5.05–4.95(m,1H),3.68-3.65(m,2H),3.63-3.53(m,5H),3.43-3.39(s,5H),3.35–3.20(m,12H),2.79-2.63(m,6H),2.43-2.39(m,2H),2.30-2.24(m,2H),1.76-1.71(m,2H),1.57-1.50(m,2H).
LCMS m/z=728.4[M+1].
化合物51
1H NMR(400MHz,DMSO-d6)δ9.82(br,1H),7.82(d,1H),7.27–6.94(m,6H),6.86(d,1H),6.55(s,1H),6.50(d,1H),4.83(dd,1H),4.03-3.99(m,2H),3.94-3.88(m,2H),3.65-3.53(m,9H),3.44-3.24(m,12H),2.80-2.72(m,3H),2.70–2.60(m,3H),2.59-2.53(m,2H),2.35-2.25(m,4H),1.75-1.42(m,7H),0.57-0.52(m,2H),0.40-0.35(m,2H).
LCMS m/z=828.5[M+1]。
化合物52
1H NMR(400MHz,Methanol-D4)δ8.01-7.92(m,2H),7.38-7.25(m,5H),6.95-6.93(m,1H),6.72-6.70(d,1H),6.51-6.49(d,1H),3.92-3.91(m,2H),3.74-3.54(m,10H),3.40-3.18(m,14H),2.90-2.87(m,4H),2.63-2.61(m,2H),2.26-2.22(m,2H),1.97-1.85(m,2H).
LCMS m/z=687.4[M+1].
化合物53、化合物54
化合物53Rx=乙基;LCMS m/z=364.8[M/2+1].
化合物54Rx=环丙基;LCMS m/z=370.8[M/2+1].
化合物55
化合物56
化合物57
1H NMR(400MHz,DMSO-d6)δ10.24(br,1H),8.19(d,1H),7.82(d,1H),7.26–7.01(m,6H),6.92(d,1H),6.55(s,1H),6.49(d,1H),5.06–4.94(m,1H),4.03–3.98(m,2H),3.96–3.89(m,2H),3.67–3.48(m,10H),3.44–3.24(m,10H),3.21–3.08(m,3H),2.78–2.68(m,4H),2.62(t,2H),2.58–2.53(m,1H),2.36–2.25(m,4H),1.74–1.39(m,4H),0.53(s,2H),0.37(s,2H).
LCMS m/z=824.5[M+1].
化合物58
1H NMR(400MHz,CDCl3)δ8.62(s,1H),7.98(d,1H),7.60(s,1H),7.30–7.27(m,1H),7.24–7.12(m,3H),6.66(d,1H),6.56–6.40(m,4H),4.57–4.48(m,3H),4.09(s,3H),3.85–3.58(m,9H),3.50–3.42(m,3H),3.37(s,2H),3.27–3.16(m,6H),2.95–2.89(m,3H),2.86–2.78(m,3H),2.61–2.55(m,2H),2.13–1.95(m,6H).
LCMS m/z=701.6[M+1].
化合物59
1H NMR(400MHz,DMSO-d6)δ9.78(s,1H),7.82(d,1H),7.22–7.15(m,1H),7.14–7.04(m,3H),6.59
(d,1H),6.55(s,1H),6.41(d,1H),4.46(d,2H),4.04–3.98(m,2H),3.97–3.87(m,2H),3.70–3.45(m,9H),3.35–3.23(m,16H),2.75(t,2H),2.65–2.59(m,3H),2.58–2.54(m,2H),2.39–2.25(m,4H),1.70–1.63(m,2H),1.55–1.46(m,2H),0.57–0.49(s,2H),0.38(s,2H).
LCMS m/z=400.8[M/2+1]。
化合物60
1H NMR(400MHz,CD3OD)δ8.05(d,1H),7.89(d,1H),7.48–7.39(m,2H),7.30(d,1H),7.24–7.18(m,2H),7.04(d,1H),6.96(d,1H),6.65(d,1H),5.40(dd,1H),4.40(s,2H),3.96–3.91(m,2H),3.81–3.57(m,11H),3.46–3.39(m,2H),3.35(s,2H),3.29–3.21(m,4H),3.09(s,3H),2.96–2.90(m,2H),2.76(q,2H),2.67(t,2H),2.24(d,2H),2.00–1.85(m,2H),1.28(t,3H).
LCMS m/z=373.3[M/2+1].
化合物61
1H NMR(400MHz,CDCl3)δ8.00(d,1H),7.22–7.05(m,4H),6.54(d,1H),6.46(d,1H),6.41(s,1H),6.27–6.19(m,1H),4.54(s,2H),3.81–3.76(m,2H),3.71(dd,2H),3.61–3.55(m,2H),3.50(s,2H),3.48–3.44(m,3H),3.39–3.27(m,5H),2.88–2.68(m,8H),2.62–2.51(m,4H),2.43(t,2H),1.91(d,2H),1.68–1.60(m,2H),1.30–1.21(m,3H),1.11(t,2H),1.06(t,1H).
LCMS m/z=358.3[M/2+1].
化合物62
1H NMR(400MHz,DMSO-d6)δ9.77(s,1H),7.82(d,1H),7.18(s,1H),7.13–7.02(m,3H),6.60(d,1H),6.55(s,1H),6.42(d,1H),4.46(d,2H),4.04–3.98(m,2H),3.96–3.86(m,2H),3.68–3.47(m,9H),3.44–3.19(m,12H),2.76(t,2H),2.66–2.60(m,3H),2.60–2.53(m,2H),2.37–2.23(m,4H),1.71–1.62(m,2H),1.56–1.44(m,2H),0.54(s,2H),0.37(s,2H).
LCMS m/z=386.8[M/2+1].
化合物63
1H NMR(400MHz,CD3OD)δ8.43(d,1H),7.76(s,1H),7.52–7.40(m,2H),7.33(d,1H),7.25(s,1H),7.07(d,1H),6.68(d,2H),5.44(d,1H),4.42(s,2H),4.31(dd,1H),4.13–4.00(m,4H),3.87–3.51(m,11H),3.47–3.39(m,2H),3.32–3.17(m,5H),2.98(t,2H),2.76(t,2H),2.31–2.17(m,2H),1.97–1.89(m,2H),1.89–1.75(m,4H),1.68–1.61(m,2H),1.59–1.47(m,2H),1.37–1.28(m,2H),0.66–0.57(m,2H),0.42(s,2H).
LCMS m/z=421.8[M/2+1].
化合物64
1H NMR(400MHz,DMSO-d6)δ9.89(dd,2H),8.82(s,1H),7.83(s,1H),7.55–7.17(m,4H),6.67(d,1H),6.57(s,1H),6.50(dd,1H),4.54(s,2H),4.30–4.16(m,1H),4.14–4.06(m,1H),4.01(d,3H),3.95–3.87(m,1H),3.71–3.56(m,7H),3.54–3.40(m,3H),3.30(s,5H),3.08–3.02(m,3H),2.99–2.92(m,2H),2.87–2.76(m,4H),2.60(t,2H),2.03–1.75(m,4H),1.35(d,2H),1.12(d,3H),1.05(d,1H),0.53(d,2H),0.37(d,2H).
LCMS m/z=393.8[M/2+1].
化合物65
1H NMR(400MHz,DMSO-d6)δ10.24(br,1H),8.18(dd,1H),7.82(d,1H),7.19(s,2H),7.06(dt,2H),6.93(dd,1H),6.55(s,1H),6.49(dd,1H),5.08–5.01(m,1H),4.01(d,2H),3.96–3.87(m,2H),3.65–3.54(m,7H),3.49(s,2H),3.43(s,1H),3.35–3.24(m,6H),2.84–2.65(m,7H),2.40–2.27(m,4H),1.71–1.63(m,2H),1.55–1.44(m,2H),1.03(t,3H),0.95(t,1H),0.56–0.49(m,2H),0.37(s,2H).
LCMS m/z=802.5[M+1].
化合物66
1H NMR(400MHz,DMSO-d6)δ9.81(br,1H),7.82(d,1H),7.22–7.17(m,1H),7.15–7.06(m,3H),6.86(d,1H),6.55(s,1H),6.50(d,1H),4.87–4.78(m,1H),4.52–4.42(m,2H),4.04–3.98(m,2H),3.96–3.88(m,2H),3.67–3.60(m,4H),3.59–3.44(m,5H),3.44–3.21(m,7H),2.79–2.74(m,2H),2.71–2.63(m,5H),2.62–2.54(m,4H),2.38–2.26(m,4H),1.71–1.64(m,2H),1.56–1.46(m,2H),0.54(s,2H),0.37(s,2H).
LCMS m/z=800.5[M+1].
化合物67
1H NMR(400MHz,DMSO-d6)δ10.32(s,1H),8.29(d,1H),8.20(t,1H),7.98(dd,1H),7.32–7.27(m,1H),7.21–7.07(m,6H),6.95(dd,1H),6.50(dd,1H),5.21–5.07(m,1H),3.77–3.71(m,2H),3.65–3.50(m,5H),3.47–3.41(m,3H),3.13–3.08(m,2H),3.03(s,2H),2.94(s,2H),2.91–2.82(m,4H),2.81–2.73(m,5H),2.58–2.52(m,2H),2.48–2.45(m,1H),2.29(t,2H),1.83(d,2H),1.60(t,2H).
LCMS m/z=350.3[M/2+1].
化合物68
1H NMR(400MHz,DMSO-d6)δ7.82(d,1H),7.19(s,1H),7.14–7.05(m,3H),6.85(d,1H),6.69(d,1H),6.55(s,1H),4.59–4.52(m,1H),4.04–3.98(s,2H),3.96–3.85(m,2H),3.67–3.46(m,9H),3.45–3.33(m,4H),3.30(t,4H),2.82–2.71(m,4H),2.63(qd,4H),2.39–2.25(m,4H),1.68(d,2H),1.56–1.46(s,2H),0.57–0.50(m,2H),0.38(s,2H).
LCMS m/z=388.8[M/2+1].
化合物69
1H NMR(400MHz,DMSO-d6)δ7.81(d,1H),7.21–7.16(m,1H),7.14–7.05(m,3H),6.85(d,1H),6.71–6.67(m,1H),6.55(s,1H),4.59–4.52(m,1H),4.04–3.98(m,2H),3.98–3.88(m,2H),3.70–3.46(m,9H),3.45–3.34(m,3H),3.31–3.19(m,5H),2.76(t,2H),2.69–2.57(m,4H),2.48–2.44(m,2H),2.39–2.26(m,4H),1.67(d,2H),1.56–1.45(m,2H),1.04(t,2H),0.57–0.50(m,2H),0.38(s,2H).
LCMS m/z=395.8[M/2+1].
化合物70
1H NMR(400MHz,DMSO-d6)δ10.06(br,1H),8.37(d,1H),8.17(d,1H),7.47(d,1H),7.23-7.16(m,2H),7.13(s,1H),7.10(d,2H),7.06(d,1H),6.91(d,1H),6.48(dd,1H),5.02–4.96(m,1H),3.73–3.67(m,2H),3.62–3.52(m,8H),3.42(d,2H),3.37-3.23(m,4H),2.79–2.71(m,4H),2.71–2.57(m,4H),2.53(d,1H),2.47–2.39(m,3H),2.26(t,2H),1.72(d,2H),1.60–1.52(m,2H).
LCMS m/z=372.8[M/2+1].
化合物71
将化合物70(0.25g,0.34mmol)溶于甲醇(10mL)中,加入钯碳(0.1g),反应在氢气氛中室温下搅拌4小时后。将反应液经硅藻土过滤后减压浓缩,柱层析分离(DCM:甲醇(v/v)=1:0~8:1)得到黄色固体状的化合物71.
1H NMR(400MHz,CD3OD)δ8.40(d,1H),7.54(d,1H),7.35(dd,4H),7.29(d,1H),7.03(d,1H),6.65(d,1H),6.33(dd,1H),6.07(s,1H),5.38(dd,1H),4.29(s,2H),3.91–3.85(m,2H),3.75(dd,3H),3.69(t,3H),3.50–3.31(m,7H),3.28–3.18(m,5H),3.08(s,3H),2.89(t,2H),2.66(t,2H),2.22(d,2H),1.86(t,2H),1.33–1.27(m,1H).
LCMS m/z=715.5[M+1].
化合物72
1H NMR(400MHz,DMSO-d6)δ10.49(s,1H),9.41(d,1H),8.08(dd,1H),7.23–7.17(m,1H),7.12(dd,3H),6.69(d,1H),6.52-6.47(m,1H),6.39(d,1H),4.10–3.99(m,1H),3.68–3.65(m,2H),3.64–3.60(m,2H),3.59–3.54(m,2H),3.50–3.44(m,3H),3.44–3.37(m,5H),3.30(s,6H),2.92(d,3H),2.84(d,1H),2.80–2.75(m,2H),2.72–2.67(m,1H),2.44–2.39(m,2H),2.30–2.23(m,2H),1.72(d,2H),1.57–1.50(m,2H).
LCMS m/z=360.3[M/2+1].
化合物73
1H NMR(400MHz,DMSO-d6)δ10.15(br,1H),8.39(dd,1H),8.23(dd,1H),8.17(d,1H),7.18(dt,1H),7.13–7.05(m,4H),6.90(dd,2H),6.48(dd,1H),5.02–4.97(m,1H),3.72–3.66(m,2H),3.65–3.48(m,5H),3.40(d,2H),3.35(s,2H),3.29(dd,3H),3.23–3.16(m,3H),2.79–2.73(m,4H),2.71–2.61(m,4H),2.53(d,1H),2.42–2.35(m,2H),2.28(t,2H),1.72(d,2H),1.56–1.47(m,2H).
LCMS m/z=745.4[M+1].
化合物74
1H NMR(400MHz,DMSO-d6)δ9.76(br,1H),8.08(dd,1H),7.19(dd,1H),7.15–7.04(m,3H),6.68(dd,1H),6.60(d,1H),6.52-6.46(m,1H),6.42(d,1H),4.47(s,2H),3.70–3.64(m,2H),3.64–3.54(m,4H),3.50–3.46(m,2H),3.41(s,4H),3.21–3.16(m,1H),3.15–3.10(m,1H),2.76(t,2H),2.67–2.52(m,7H),2.46–2.40(m,3H),2.27(t,2H),1.72(d,3H),1.68–1.58(m,3H),1.58–1.52(m,2H),1.51–1.42(d,4H),1.26–1.22(m,1H).
LCMS m/z=380.3[M/2+1]。
化合物75
1H NMR(400MHz,DMSO-d6)δ10.05(br,1H),8.18(d,1H),7.89(d,1H),7.22-7.16(m,1H),7.13(s,1H),7.08(dd,3H),6.91(d,1H),6.53(s,1H),6.48(d,1H),6.28(dd,1H),5.03–4.96(m,1H),3.69–3.64(m,2H),3.63–3.54(m,4H),3.41(s,4H),3.34(s,2H),3.32–3.18(m,5H),2.76(t,2H),2.72–2.57(m,4H),2.48–2.38(m,4H),2.27(t,2H),1.85–1.77(m,1H),1.73(d,2H),1.54(t,2H),1.27–1.19(m,1H),0.97-0.93(m,4H),0.76–0.71(m,2H).
LCMS m/z=753.5[M+1].
化合物76
1H NMR(400MHz,DMSO-d6)δ9.77(br,1H),8.08(dd,1H),7.23-7.16(m,1H),7.10(dd,3H),6.69(dd,1H),6.60(d,1H),6.51-6.46(m,1H),6.42(dd,1H),4.47(d,2H),3.70–3.64(m,2H),3.63–3.53(m,4H),3.50–3.46(m,2H),3.44–3.38(m,4H),3.30(dd,3H),2.91(s,1H),2.81–2.74(m,3H),2.68–2.52(m,6H),2.45–2.39(m,2H),2.27(t,2H),1.72(d,2H),1.58–1.47(m,2H),1.28–1.20(m,2H).
LCMS m/z=353.1[M/2+1].
化合物77
1H NMR(400MHz,DMSO-d6)δ9.77(br,1H),7.89(d,1H),7.23-7.16(m,1H),7.11(dd,3H),6.60(d,1H),6.54(s,1H),6.42(dd,1H),6.29(dd,1H),4.47(d,2H),3.71–3.63(m,2H),3.63–3.53(m,4H),3.45–3.38(m,4H),3.34(s,2H),3.32–3.26(m,5H),2.79–2.74(m,3H),2.67–2.60(m,3H),2.58–2.53(m,2H),2.45–2.40(m,2H),2.28(t,2H),1.84-1.78(m,1H),1.77–1.69(m,2H),1.54(t,2H),1.25–1.22(m,2H),0.97–0.93(m,2H),0.76-0.72(m,2H).
LCMS m/z=364.3[M/2+1].
化合物78
将化合物73(0.27g,0.36mmol)溶于甲醇(10mL)中,加入钯碳(0.1g),氢气氛室温搅拌4小时。反应液用硅藻土过滤后减压浓缩,柱层析分离(DCM:甲醇(v/v)=1:0~8:1)得到黄色固体状的化合物78.
1H NMR(400MHz,CD3OD)δ8.39(d,1H),7.62(d,1H),7.47(dd,1H),7.41–7.31(m,4H),7.29(d,1H),7.22(dd,1H),7.02(d,1H),6.65(d,1H),5.38(dd,1H),4.31(s,2H),3.98–3.94(m,2H),3.79–3.62(m,6H),3.43–3.32(m,5H),3.28–3.21(m,6H),3.15(s,1H),3.09(s,2H),3.00–2.75(m,3H),2.66(t,2H),2.46(d,2H),1.81(t,2H),1.33–1.26(s,1H).
LCMS m/z=357.7[M/2+1].
化合物79
1H NMR(400MHz,CD3OD)δ8.27(d,1H),7.43–7.30(m,6H),7.00(s,1H),6.86(d,1H),5.17(dd,1H),4.55(d,1H),4.29(s,3H),4.14(dd,1H),3.86–3.81(m,3H),3.77–3.65(m,7H),3.63–3.52(dd,6H),3.50(dd,1H),3.34(s,1H),2.90(s,3H),2.63(dt,3H),2.23(d,3H),2.10–1.90(m,1H),1.77(t,3H).
LCMS m/z=386.2[M/2+1].
化合物80
化合物81
化合物82
化合物83
化合物84
化合物85
化合物86
化合物87
化合物88
化合物89
化合物90
化合物91
1H NMR(400MHz,CD3OD)δ8.29(d,2H),7.25–7.05(m,4H),6.67(d,1H),6.56(t,1H),6.44(d,1H),4.52(d,2H),4.33–4.07(m,1H),3.64-3.74(m,10H),3.48(d,2H),3.33(d,2H),,2.73-2.85(m,8H),2.56-2.65(m,2H),2.50(s,2H),2.43(d,2H),1.72-1.83(m,6H),1.66–1.43(m,6H).
LCMS m/z=371.7[M/2+1].
化合物92
1H NMR(400MHz,CD3OD)δ7.96-7.95(d,1H),7.20-7.13(m,4H),6.68-6.59(m,3H),6.45-6.43(d,1H),4.54-4.50(d,2H),3.78-3.68(m,6H),3.52-3.42(m,6H),2.84(s,2H),2.75-2.65(m,10H),2.59-2.54(m,3H),2.45-2.41(m,2H),1.92-1.89(m,2H),1.64-1.61(m,2H),1.23-1.21(m,6H).
LCMS m/z=365.3[M/2+1].
化合物93
1H NMR(400MHz,CD3OD)δ8.39-8.35(m,1H),8.06-8.05(d,1H),7.53-7.51(m,1H),7.25-7.13(m,5H),6.98-6.96(d,1H),6.77-6.75(d,1H),6.64-6.61(m,2H),5.21-5.18(m,1H),3.76-3.64(m,7H),3.48-3.36(m,8H),2.86-2.76(m,6H),2.58-2.53(m,4H),2.48-2.40(m,2H),1.90-1.88(m,2H),1.54-1.40(m,5H),1.35-1.26(m,2H),0.94-0.88(m,3H).
LCMS m/z=741.3[M+1].
化合物94
1H NMR(400MHz,CDCl3)δ8.00-7.98(m,1H),7.53-7.48(m,2H),7.39-7.35(m,2H),7.16-7.07(m,2H),6.98-6.95(m,2H),6.89-6.80(m,2H),6.52-6.45(m,1H),5.20-5.12(m,1H),3.75-3.14(m,15H),2.94-2.59(m,16H),1.92-1.89(m,2H),1.52-1.43(m,2H),1.29-1.26(m,4H),0.92-0.88(m,3H)
LCMS m/z=807.5[M+1].
化合物95
1H NMR(400MHz,CD3OD)δ7.74(d,1H),7.30–7.19(m,5H),7.12(s,1H),6.80(d,1H),6.57(d,1H),6.37(d,1H),4.33(s,2H),4.14(s,2H),3.83–3.80(m,2H),3.58–3.48(m,11H),3.42(t,3H),3.15–3.08(m,2H),3.02(t,2H),2.77(t,2H),2.68(t,2H),2.43(t,2H),2.35(s,3H),2.10(d,2H),1.98(dd,2H),1.81(d,2H),1.68(d,2H),1.59-1.46(m,5H).
LCMS m/z=378.3[M/2+1].
化合物96
1H NMR(400MHz,CD3OD)δ8.39-8.35(m,1H),7.96-7.95(d,1H),7.22-7.16(m,5H),6.98-6.96(d,1H),6.65-6.58(m,3H),5.20-5.17(m,1H),3.78-3.64(m,6H),3.51-3.35(m,9H),2.86-2.77(m,6H),2.60-2.52(m,4H),2.48-2.42(m,2H),1.93-1.89(m,2H),1.68-1.62(m,2H),1.40-1.38(m,4H),1.23-1.21(m,6H).
LCMS m/z=378.3[M/2+1].
化合物97
1H NMR(400MHz,CD3OD)δ7.97-7.96(d,1H),7.30-7.23(m,5H),6.75-6.72(d,1H),6.65-6.61(m,2H),
6.50-6.48(m,1H),4.54-4.50(m,2H),3.81-3.71(m,7H),3.52-3.43(m,6H),3.15-3.07(m,3H),2.87-2.67(m,10H),2.15-1.26(m,16H),1.24-1.22(m,6H).
LCMS m/z=783.5[M+1].
化合物98
化合物99
化合物100
化合物101
化合物102
化合物103
化合物104
化合物105
化合物106
化合物107
化合物108
化合物109
化合物110
化合物111
化合物112
1H NMR(400MHz,CDCl3)δ7.46(d,1H),7.21–7.04(m,4H),6.69(d,1H),6.54(s,1H),6.24(d,1H),4.54(d,2H),4.09(s,1H),3.98–3.88(m,3H),3.81–3.52(m,9H),3.47(s,3H),3.38(d,1H),3.31–3.23(m,1H),3.22–3.17(m,1H),2.91–2.65(m,8H),2.62–2.46(m,5H),2.38(s,1H),1.83(s,2H),1.71(s,1H),1.61-1.46(m,3H),1.32–1.18(m,3H),0.63(s,2H),0.35(s,2H).
LCMS m/z=393.8[M/2+1]
化合物113
1H NMR(400MHz,CD3OD)δ8.43(d,1H),7.93(d,1H),7.43–7.35(m,4H),7.33(d,1H),7.17(s,1H),7.07(d,1H),7.00(d,1H),6.69(d,1H),5.44(dd,1H),4.32(d,3H),3.99–3.94(m,2H),3.79(t,2H),3.71(dt,5H),3.61(d,3H),3.38(s,2H),3.27(d,4H),3.04(dt,1H),2.93(t,2H),2.77(t,2H),2.30–2.22(m,2H),2.06(d,1H),1.93(s,4H),1.79(s,2H),1.71–1.50(m,5H),1.38–1.25(m,7H).
LCMS m/z=398.3[M/2+1].
化合物114
化合物115
1H NMR(400MHz,DMSO-d6)δ10.14(br,1H),8.17(d,1H),7.94(d,1H),7.20(dd,2H),7.08–7.02(m,2H),6.91(d,1H),6.65(s,1H),6.48(dd,2H),5.02–4.97(m,1H),3.69–3.64(m,2H),3.62–3.54(m,4H),3.49(s,2H),3.43–3.39(m,2H),3.38–3.10(m,12H),2.81(t,2H),2.72–2.61(m,4H),2.32(t,2H),1.74(d,2H),1.53(t,2H),1.23(d,3H).
LCMS m/z=366.3[M/2+1].
化合物116
1H NMR(400MHz,DMSO-d6)δ8.17(dd,1H),7.81(d,1H),7.20(d,2H),7.05(t,2H),6.91(d,1H),6.56(s,1H),6.48(dd,1H),5.00(t,1H),4.01(d,2H),3.91(s,2H),3.70–3.52(m,8H),3.48(d,2H),3.43(s,1H),3.30(t,2H),2.91(s,1H),2.80(s,2H),2.77(s,1H),2.72–2.63(m,4H),2.53(d,1H),2.47(s,1H),2.32(s,4H),1.66(s,2H),1.50(s,2H),1.23(d,2H),0.53(d,2H),0.37(s,2H).
LCMS m/z=394.8[M/2+1].
化合物117
1H NMR(400MHz,CD3OD)δ8.39-8.34(m,1H),8.26-8.25(m,1H),7.19-7.13(m,5H),7.00-6.98(m,2H),6.81-6.80(d,1H),6.65-6.61(m,1H),5.21-5.16(m,1H),3.71-3.64(m,6H),3.55-3.46(m,8H),3.02(s,2H),2.84-2.75(m,7H),2.63-2.60(m,4H),2.46-2.41(m,2H),1.90-1.86(m,2H),1.63-1.61(m,2H).
LCMS m/z=384.3[M/2+1].
化合物118
1H NMR(400MHz,CD3OD)δ7.72(s,1H),7.23(dd,2H),7.08(t,1H),6.74(d,1H),6.62(d,1H),6.50(d,1H),4.62(s,1H),4.30–4.20(m,1H),4.03(d,2H),3.98(d,2H),3.80–3.57(m,11H),3.51(t,2H),3.12(dt,3H),2.95–2.85(m,4H),2.70–2.64(m,3H),2.56(d,3H),1.88(s,4H),1.81–1.55(m,8H),1.51(dd,2H),0.61(d,2H),0.40(d,2H).
LCMS m/z=415.8[M/2+1]。
化合物119
1H NMR(400MHz,DMSO-d6)δ10.21(br,1H),8.29(d,1H),8.18(dd,1H),7.18(dd,1H),7.13(s,1H),7.11–7.05(m,3H),6.92(dd,1H),6.83(d,1H),6.49(dd,1H),5.15(s,1H),5.03(dd,1H),3.70(d,2H),3.67–3.53(m,9H),3.41(d,2H),3.38–3.29(m,3H),2.92(s,1H),2.79–2.66(m,8H),2.64(d,1H),2.35(s,4H),1.67(d,2H),1.54(d,2H),1.36(s,6H).
LCMS m/z=379.8[M/2+1].
化合物120
1H NMR(400MHz,DMSO-d6)δ10.26(br,1H),8.42(d,1H),8.17(dd,1H),8.13(dd,2H),7.54–7.49(m,3H),7.21–7.16(m,2H),7.13–7.05(m,4H),6.92(dd,1H),6.48(dd,1H),5.02(s,1H),3.81(d,2H),3.76–3.67(m,4H),3.57(dt,5H),3.42(d,3H),3.30(t,6H),2.91(s,1H),2.80–2.65(m,5H),2.42–2.29(m,4H),1.72(d,2H),1.57(t,2H).
LCMS m/z=388.8[M/2+1]。
化合物121
化合物122
化合物123
化合物124
1H NMR(400MHz,DMSO-d6)δ9.81(br,1H),7.97(d,1H),7.19(d,1H),7.16–7.06(m,3H),6.87(d,1H),6.66(s,1H),6.51(dd,2H),4.83(d,1H),4.47(s,2H),3.68(s,2H),3.58(dd,4H),3.42(s,4H),3.37–3.17(m,12H),2.77(t,3H),2.65(s,3H),2.56(d,3H),2.43(d,2H),2.28(t,2H),1.75(d,3H),1.68–1.43(m,9H).
LCMS m/z=400.3[M/2+1]。
化合物125
1H NMR(400MHz,CD3OD)δ8.38-8.34(m,1H),7.88-7.86(d,1H),7.22-7.14(m,5H),6.98-6.96(d,1H),6.65-6.62(m,1H),6.52(s,1H),6.33-6.31(m,1H),5.20-5.16(m,1H),3.77-3.65(m,6H),3.53-3.40(m,8H),3.02(s,2H),2.84-2.75(m,7H),2.58-2.55(m,4H),2.46-2.41(m,2H),1.88-1.84(m,2H),1.67-1.64(m,2H),
1.04-1.02(m,2H),0.77-0.74(m,2H).
LCMS m/z=370.3[M/2+1]
化合物126
1H NMR(400MHz,CD3OD)δ8.38-8.34(m,1H),7.89-7.88(d,1H),7.22-7.11(m,5H),6.98-6.96(d,1H),6.81-6.75(m,2H),6.63-6.61(m,1H),5.21-5.16(m,1H),3.71-3.64(m,7H),3.52-3.26(m,6H),3.02(s,2H),2.84-2.75(m,8H),2.58-2.55(m,4H),2.39-2.36(m,2H),1.88-1.84(m,2H),1.64-1.56(m,2H).
LCMS m/z=733.3[M+1].
化合物127
1H NMR(400MHz,DMSO-d6)δ9.83(br,1H),7.96(d,1H),7.21–7.17(m,1H),7.15–7.07(m,3H),6.87(d,1H),6.66(s,1H),6.54–6.46(m,2H),4.86(s,1H),4.52–4.44(m,2H),3.67(d,2H),3.59(dd,5H),3.43(s,5H),3.36–3.17(m,8H),2.77(t,2H),2.68(d,4H),2.58(t,2H),2.43(d,2H),2.29(t,2H),1.75(d,3H),1.65(s,3H),1.53(dd,7H).
LCMS m/z=393.4[M/2+1]。
化合物128
1H NMR(400MHz,CD3OD)δ8.26-8.25(d,1H),7.23-7.13(m,4H),6.96-6.91(m,2H),6.81-6.80(d,1H),6.72-6.70(m,1H),4.76-4.72(m,1H),3.78-3.66(m,6H),3.57-3.53(m,8H),3.01(s,2H),2.84-2.75(m,7H),2.63-2.57(m,4H),2.45-2.40(m,2H),1.90-1.86(m,2H),1.63-1.61(m,2H).
LCMS m/z=387.3[M/2+1].
化合物129
1H NMR(400MHz,CD3OD)δ8.39-8.34(m,1H),8.04-8.02(m,1H),7.21-7.13(m,5H),6.98-6.93(d,1H),6.64-6.62(m,1H),6.54-6.40(m,2H),5.21-5.16(m,1H),3.75-3.66(m,6H),3.53-3.44(m,8H),3.02(s,2H),2.84-2.75(m,7H),2.63-2.60(m,4H),2.46-2.41(m,2H),1.89-1.85(m,2H),1.65-1.60(m,2H).
LCMS m/z=718.4[M+1].
化合物130
化合物131
化合物132
化合物133
化合物134
LCMS m/z=379.8[M+1]。
化合物135
化合物136
1H NMR(400MHz,CD3OD)δ8.35(dd,1H),8.28(d,2H),7.25–7.05(m,5H),6.96(d,1H),6.62(dd,1H),6.55(t,1H),5.16-5.21(m,1H),3.77–3.62(m,10H),3.56–3.41(m,4H),3.01(s,2H),2.92–2.73(m,7H),2.67–2.48(m,4H),2.43(t,2H),1.81(d,2H),1.70–1.53(m,2H).
LCMS m/z=351.0[M/2+1].
化合物137:7-[(1R)-2-[2-[3-氟-5-[[4-(4-甲基-2-吡啶基)-1-氧-4,9-二氮杂螺[5.5]十一烷-9-基]甲基]苯基]乙胺基]-1-羟基-乙基]-4-羟基-3H-1,3-苯并噻唑-2-酮
7-[(1R)-2-[2-[3-fluoro-5-[[4-(4-methyl-2-pyridyl)-1-oxa-4,9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethylamino]-1-hydroxy-ethyl]-4-hydroxy-3H-1,3-benzothiazol-2-one
第一步:2-[3-氟-5-[[4-(4-甲基-2-吡啶基)-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-基]甲基]苯基]乙醇(137C)
2-[3-fluoro-5-[[4-(4-methyl-2-pyridyl)-1-oxa-4,9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethanol
将137A(0.61g,2.5mmol)溶于乙腈(30mL)中,加入137B(0.63g,2.7mmol),碳酸钾(0.68g,4.9mmol)
和水(1mL),50℃反应2.5小时。加入水(100mL),用DCM(100mL×2)萃取,合并后的有机相用饱和食盐水洗(100mL),无水硫酸钠干燥,减压浓缩后柱层析分离(DCM/甲醇(v/v)=15:1)得到黄色液体状的137C(0.80g,产率81%)。
第二步:9-[[3-(2-溴乙基)-5-氟-苯基]甲基]-4-(4-甲基-2-吡啶基)-1-氧杂-4,9-二氮杂螺[5.5]十一烷(137D)
9-[[3-(2-bromoethyl)-5-fluoro-phenyl]methyl]-4-(4-methyl-2-pyridyl)-1-oxa-4,9-diazaspiro[5.5]undecane
将137C(0.80g,2.0mmol)溶于DCM(30mL)中,加入咪唑(0.2g,3.0mmol),四溴化碳(0.80g,2.4mmol),三苯基膦(0.79g,3.0mmol),室温反应5小时。加入水(100mL),萃取,水相用DCM(50mL×2)萃取,合并后的有机相用饱和食盐水(100mL)洗涤,无水硫酸钠干燥,减压浓缩后柱层析分离(DCM/甲醇(v/v)=20:1)得到黄色液体状的137D(0.45g,产率49%)。
第三步:7-[(1R)-1-[叔丁基(二甲基)硅基]氧-2-[2-[3-氟-5-[[4-(4-甲基-2-吡啶基)-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-基]甲基]苯基]乙胺基]乙基]-4-羟基-3H-1,3-苯并噻唑-2-酮(137E)
7-[(1R)-1-[tert-butyl(dimethyl)silyl]oxy-2-[2-[3-fluoro-5-[[4-(4-methyl-2-pyridyl)-1-oxa-4,9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethylamino]ethyl]-4-hydroxy-3H-1,3-benzothiazol-2-one
将137D(0.45g,0.97mmol)溶于DMF(30mL)中,加入7-[(1R)-2-氨基-1-[叔丁基(二甲基)硅基]氧乙基]-4-羟基-3H-1,3-苯并噻唑-2-酮(0.30g,0.88mmol),二异丙基乙胺(0.72g,5.6mmol),50℃反应40小时,反应液直接减压浓缩后柱层析分离(DCM/甲醇(v/v)=10:1)得到黄色固体状的137E(0.12g,产率19%)。
第四步:7-[(1R)-2-[2-[3-氟-5-[[4-(4-甲基-2-吡啶基)-1-氧杂-4,9-二氮杂螺[5.5]十一烷-9-基]甲基]苯基]乙胺基]-1-羟基-乙基]-4-羟基-3H-1,3-苯并噻唑-2-酮(化合物137)
7-[(1R)-2-[2-[3-fluoro-5-[[4-(4-methyl-2-pyridyl)-1-oxa-4,9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethylamino]-1-hydroxy-ethyl]-4-hydroxy-3H-1,3-benzothiazol-2-one
将137E(0.12g,0.17mmol)溶于THF(10mL)中,加入三乙胺三氢氟酸(0.5g,3.0mmol),室温搅拌12小时。依次滴加饱和碳酸氢钠水溶液(50mL),甲醇(5mL)淬灭反应,残余物用DCM(100mL×2)萃取,合并后的有机相用无水硫酸钠干燥,减压浓缩后柱层析分离(DCM:甲醇(v/v)=1:0~8:1)得到黄色固体状的化合物137(0.03g,产率30%)。
LCMS m/z=304.7[M/2+1].
化合物138
化合物139
1H NMR(400MHz,DMSO-d6)δ8.18(d,1H),7.22-7.17(td,1H),7.14(s,1H),7.08(dd,3H),6.91(d,1H),6.88–6.74(m,4H),6.48(d,1H),5.05–4.96(m,1H),3.75–3.69(m,2H),3.67(s,3H),3.63-3.54(m,4H),3.42(d,2H),3.31(dd,2H),2.93–2.88(m,3H),2.80(s,2H),2.78–2.75(m,3H),2.73–2.62(m,4H),2.54(d,1H),2.46(dd,3H),2.28(t,2H),1.84(d,2H),1.58(t,2H),1.23(s,2H).
LCMS m/z=364.8[M/2+1].
化合物140
化合物141
1H NMR(400MHz,DMSO-d6)δ8.40(dd,1H),8.17(d,1H),8.05(dd,1H),7.23-7.16(m,1H),7.14–7.04(m,4H),6.94–6.89(m,2H),6.48(dd,1H),5.00(dd,1H),3.77–3.72(m,2H),3.63–3.50(m,7H),3.47(s,2H),3.41(d,2H),3.38–3.25(m,4H),2.80–2.72(m,4H),2.71–2.57(m,4H),2.54(d,1H),2.47–2.42(m,2H),2.26(t,2H),1.84–1.76(m,2H),1.55(t,2H).
LCMS m/z=362.8[M/2+1]。
化合物142
1H NMR(400MHz,DMSO-d6)δ10.22(s,1H),8.17(d,1H),7.39(dd,1H),7.31–7.26(m,1H),7.22-7.16(m,1H),7.16–7.00(m,6H),6.91(d,1H),6.52–6.43(m,1H),5.00(dd,1H),3.79–3.72(m,2H),3.62-3.54(m,4H),3.41(d,2H),3.35-3.25(m,4H),2.92–2.88(m,3H),2.76(d,5H),2.73–2.60(m,4H),2.54(d,1H),2.48–2.40(m,3H),2.30(t,2H),1.92(d,2H),1.61(t,2H).
LCMS m/z=366.8[M/2+1]。
化合物143
化合物144
化合物145
化合物146
将化合物145(0.41g,0.5mmol)溶于甲醇(10mL)中,加入10%钯碳(100mg),氢气氛室温搅拌2小时。反应液减压浓缩反应液后柱层析分离(二氯甲烷/甲醇(v/v)=4:1)得到黄色固体状的化合物146(0.37g,产率50%).
化合物147
化合物148
采用与化合物137相似的合成方法(相同溶剂,温度,试剂和投料比例),将137B用代替,将7-[(1R)-2-氨基-1-[叔丁基(二甲基)硅基]氧乙基]-4-羟基-3H-1,3-苯并噻唑-2-酮用代替。
生物测试例
测试例1:对人毒蕈碱M3受体的抑制活性
稳定表达人毒蕈碱受体3(hM3)和apo-Aequorin的CHO细胞(PerkinElmer,ES-212-AF)培养于含10%胎牛血清(FBS)(Gibico 10099-141),400μg/mL G418(sigma G5013)和250μg/mL Zeocin(invivogenant-zn-5p)的Ham’S F12培养基(Invitrogen 12500-062)中,在37℃,5%CO2条件下培养,达到90-100%融合。以PBS/5mM EDTA冲洗分离细胞,离心收集,以含0.1%BSA(BOVOGEN BSAS 100)无酚红Ham’s F12培养基(Invitrogen 11039-021)重悬细胞并计数,调整细胞浓度至1×106cells/mL。将15ml细胞悬液加入50mL离心管,加入Coelenterazine-h(promegaS2011)至终浓度为5μM。用锡纸包裹避光,于旋转摇床20℃下孵育4小时。再以0.1%BSA/无酚红Ham’s F12培养基稀释细胞至终浓度为5.0×105cells/mL,将细胞置于旋转摇床上低速转动,室温下孵育至少1小时。实施例化合物用DMSO配制为10mM母液,0.1%BSA/无酚红Ham’s F12培养基梯度稀释(log(M):-7,-8,-9,-10,-11),加入96孔板,每孔50μL。每孔再加入50μL细胞悬液(25000细胞/孔),室温孵育15分钟。将96孔板放入酶标仪(Perkin Elmer,Envision),以酶标仪加样器每孔加入50μL氯化乙酰胆碱(Sigma A6625)溶液,其浓度为112.92nM(hM3),记录发光20秒,使用origin7.5计算和分析IC50。本发明化合物人毒蕈碱受体的抑制活性通过以上的实验进行测定,测得的IC50值见下表1。表1测试化合物对人毒蕈碱M3受体的抑制活性结果
结论:本发明对人毒蕈碱M3受体有显著抑制活性。
测试例2:对人肾上腺素能β2受体的激动活性
实施例化合物对人肾上腺素能受体的激动活性通过LANCE Ultra cAMP Assay测定。
稳定表达人肾上腺素能受体(hβ2)的CHO细胞(PerkinElmer,ES-034-CF)培养于含10%胎牛血清(FBS)(Gibico 10099-141)和250μg/mL Zeocin(InvivoGen ant-zn-5p)的MEM-alpha培养基(Invitrogen12561-056),在37℃,5%CO2条件下培养,达到90-100%融合后用LANCE Ultra cAMP Assay试剂盒(PerkinElmer TRF0263)检测实施例对cAMP的激动作用。以PBS/5mM EDTA分离细胞,离心收集,用Stimulation Buffer(1x HBSS,5mM HEPES,0.5mM IBMX,0.1%BSA,PH7.4)重悬细胞,调整细胞浓度至6×105cells/ml。实施例化合物用DMSO配制为10mM母液,以Stimulation Buffer梯度稀释后以每孔5μl加入384孔板。每孔再加入5μL细胞悬液(3000细胞/孔),室温孵育30分钟后,每孔加入5μl 4x Eu-cAMP tracer工作溶液,然后每孔加入5μl 4x Ulight-anti-cAMP工作溶液,并在室温下孵育1小时。384孔板用酶标仪(Perkin Elmer,Envision)检测TR-FRET,使用origin7.5计算和分析EC50。本发明化合物对人肾上腺素能受体的激动活性通过以上的实验进行测定,测得的EC50值见表2:
表2测试化合物对人肾上腺素能β2受体的激动活性结果
编号 | hβ2受体EC50(nM) | 编号 | hβ2受体EC50(nM) |
1 | 1.04 | 76 | 1.88 |
2 | 0.44 | 77 | 3.09 |
3 | 0.73 | 78 | 1.76 |
4 | 1.21 | 80 | 0.64 |
5 | 0.265 | 81 | 0.29 |
6 | 4.49 | 82 | 0.62 |
7 | 0.56 | 84 | 1.57 |
9 | 7.19 | 85 | 0.13 |
10 | 0.39 | 86 | 0.05 |
11 | 0.73 | 90 | 4.14 |
12 | 0.91 | 91 | 5.99 |
13 | 0.67 | 92 | 4.14 |
16 | 1.78 | 93 | 2.5 |
18 | 0.72 | 96 | 0.96 |
19 | 0.67 | 98 | 0.52 |
20 | 0.28 | 99 | 0.24 |
21 | 0.25 | 100 | 0.24 |
22 | 0.16 | 101 | 0.53 |
23 | 0.14 | 102 | 2.33 |
25 | 0.5 | 103 | 2.27 |
26 | 1.26 | 104 | 8.18 |
28 | 0.77 | 105 | 9.09 |
29 | 3.29 | 106 | 0.3 |
32 | 2.53 | 107 | 1.53 |
37 | 1.05 | 108 | 0.98 |
38 | 0.79 | 109 | 8.32 |
39 | 0.51 | 110 | 0.9 |
43 | 0.18 | 111 | 7.3 |
44 | 1.98 | 114 | 2.79 |
45 | 1.58 | 115 | 0.72 |
47 | 2.73 | 116 | 0.96 |
52 | 0.28 | 117 | 0.27 |
53 | 0.66 | 118 | 3.06 |
54 | 0.76 | 119 | 1.04 |
55 | 0.14 | 120 | 0.71 |
56 | 0.45 | 121 | 0.22 |
57 | 7.79 | 122 | 0.88 |
58 | 1.91 | 123 | 0.97 |
59 | 4.71 | 124 | 5.13 |
60 | 0.12 | 125 | 0.18 |
61 | 2.25 | 126 | 0.19 |
62 | 2.92 | 127 | 0.92 |
64 | 5.19 | 128 | 0.13 |
65 | 5.02 | 129 | 0.09 |
66 | 3.87 | 130 | 0.12 |
67 | 0.29 | 131 | 0.36 |
68 | 0.56 | 132 | 0.3 |
69 | 1.03 | 133 | 0.87 |
70 | 0.14 | 134 | 0.53 |
71 | 0.32 | 135 | 0.29 |
72 | 2.29 | 136 | 0.85 |
73 | 0.04 | 137 | 0.9 |
74 | 4.61 | 138 | 5.6 |
75 | 0.8 |
结论:本发明对β2肾上腺素能受体有显著激动活性。
测试例3:乙酰甲胆碱诱导的豚鼠支气管收缩抑制作用
8周龄全雄豚鼠购置于维通利华,适应3天后开始实验。待测用83%无水乙醇+17%吐温80配制成0.6mM储备液。于给药前用水稀释成所需浓度。给药前,使用小动物麻醉机(Matrx;VME2)给予5%异氟烷麻醉动物,麻醉时间为1.5~2分钟。待豚鼠麻醉后,将豚鼠固定于气管插管操作平台上,使用大鼠液体气溶胶给药套装(penn-century;MSA-250-R)气管内给药,每只豚鼠给药体积250μl。给药后,于4小时,24小时,使用全体积描计仪(DSI;GS220A12-R7B)测量豚鼠增强呼气间歇(enhanced pause;PenH)值。雾化给予3mg/ml乙酰甲胆碱(Mch),雾化时间36秒,记录时间7分钟。计算PenH平均值。(参考文献J Pharmacol Exp Ther 345:260-270.)。实验结果见表3。
PenH计算公式:PenH=PEP/PIP*Pause;Pause=(Te-Tr)/Tr
Te:呼气相时间(s)
Tr:松弛相时间(s)
PEP:呼气峰流速(ml/s)
PIP:吸气峰流速(ml/s)
表3化合物对乙酰甲胆碱诱导的豚鼠支气管收缩抑制作用结果
“-”代表未测试。
结论:本发明化合物对乙酰甲胆碱诱导的豚鼠支气管收缩具有明显的抑制,且在给药24小时后,部分化合物仍具有良好的支气管收缩抑制效果。
Claims (14)
- 一种通式(I)所示的化合物或其立体异构体、水合物、代谢产物、溶剂化物、药学上可接受的盐、共晶或前药:其中:R1选自C6-12碳环或者5至12元的杂环,所述的碳环或者杂环任选进一步被0至5个R1a取代,且所述的杂环含有1至4个选自N、O或S的杂原子;R1a选自F、Cl、Br、I、-(=O)、硝基、氰基、羟基、羧基、-C(=O)OC1-6烷基、氨基、C1-6烷基、C2-6炔基、C1-6烷氧基、C3-12碳环、5至12元的杂环、-O-C3-12碳环、-NH-C3-12碳环或-CH2-C3-6环烷基,所述的羟基、氨基、烷基、炔基、烷氧基、碳环、杂环或环烷基任选进一步被0至4个选自F、Cl、Br、I、氰基、羟基、C1-6烷基、C2-6炔基、C1-4烷氧基、C3-6环烷基或C3-10碳环或5至10元的杂环的取代基所取代,所述的杂环含有1至4个选自N、O或S的杂原子;W选自键或-C(=O)-;条件是W选自-C(=O)-时,R1被1至5个R1a取代,且至少有1个R1a选自C2-6炔基或-CH2-C3-6环烷基;R2、R3、R4、R5各自独立的选自C1-6亚烷基,所述的亚烷基任选进一步被0至4个选自F、Cl、Br、I、氰基、羟基、C1-6烷基、C1-6烷氧基、苯基或苯基-C1-4亚烷基的取代基所取代;A选自C3-12碳环或5至12元杂环,所述的碳环或杂环任选进一步被0至5个F、Cl、Br、I、氰基、羟基、氨基、C1-6烷基、C2-6炔基、C1-4烷氧基或C3-6环烷基的取代基取代,且所述的杂环含有1至3个选自N、O或S的杂原子;X1和X2各自独立的选自键、-O-、-C(=O)-、-C(=O)O-、-OC(=O)-、-S-、-S(=O)-、-S(=O)2-、-C(=O)NRx-、-NRxC(=O)-、-OC(=O)NRx-、-NRxC(=O)O-、-NRxC(=O)NRx、-NRxS(=O)2-、-S(=O)2NRx-、-NRxS(=O)2NRx或-NRx-;Rx各自独立选自H、C1-6烷基或C3-8环烷基,所述的烷基或环烷基任选进一步被0至5个选自F、Cl、Br、I、C1-4烷基、C3-6环烷基或C1-4烷氧基的取代基所取代;作为选择R3-X1-R4-X2-R5可为亚乙基;R6、R7各自独立的选自H或C1-4烷基;R8选自H或OH;
- 根据权利要求2所述的化合物或其立体异构体、水合物、代谢产物、溶剂化物、药学上可接受的盐、共晶或前药:R1选自C6-10碳环或者5至10元的杂环,所述的碳环或者杂环任选进一步被0至5个R1a取代,且所述的杂环含有1至4个选自N、O或S的杂原子;R1a选自F、Cl、Br、-(=O)、硝基、氰基、羟基、氨基、羧基、-C(=O)OC1-4烷基、C1-4烷基、C2-4炔基、C1-4烷氧基、C3-10碳环、5至10元的杂环、-O-C3-10碳环、-NH-C3-10碳环或-CH2-C3-6环烷基,所述的羟基、氨基、烷基、炔基、烷氧基、碳环、杂环或环烷基任选进一步被0至4个选自F、Cl、Br、氰基、羟基、C1-4烷基、C1-4烷氧基、C3-6环烷基、C3-10碳环或5至10元的杂环的取代基所取代,所述的杂环含有1至4个选自N、O或S的杂原子;W选自键或-C(=O)-;条件是W选自-C(=O)-时,R1被1至5个R1a取代,且至少有1个R1a选自C2-4炔基或-CH2-C3-6环烷基;R2、R5各自独立的选自C1-4亚烷基,所述的亚烷基任选进一步被0至4个选自F、甲基、乙基、甲氧基或乙氧基的取代基所取代;R3、R4各自独立的选自C1-4亚烷基,所述的亚烷基任选进一步被0至4个选自F、甲基、乙基、丙基、异丙基、丁基、甲氧基、乙氧基、苯基、苯基-亚甲基、苯基-亚乙基、苯基-亚丙基或苯基-亚丁基的取代基所取代;A选自C6-10碳环或5至10元杂环,所述的碳环或杂环任选进一步被0至5个F、Cl、Br、I、氰基、羟基、氨基、C1-6烷基、C2-6炔基、C1-4烷氧基或C3-6环烷基的取代基取代,且所述的杂环含有1至3个选自N、O或S的杂原子;X1和X2各自独立的选自键、-O-、-C(=O)-、-C(=O)O-、-OC(=O)-、-S-、-S(=O)-、-S(=O)2-、-C(=O)NRx-、-NRxC(=O)-、-OC(=O)NRx-、-NRxC(=O)O-、-NRxC(=O)NRx、-NRxS(=O)2-、-S(=O)2NRx-、-NRxS(=O)2NRx或-NRx-;Rx各自独立选自H、C1-6烷基或C3-8环烷基,所述的烷基或环烷基任选进一步被0至5个选自F、Cl、Br、I、甲基、乙基、环丙基、环丁基、环戊基、环己基、甲氧基或乙氧基的取代基所取代;作为选择R3-X1-R4-X2-R5可为亚乙基;R6、R7各自独立的选自H或C1-4烷基。
- 根据权利要求3所述的化合物或其立体异构体、水合物、代谢产物、溶剂化物、药学上可接受的盐、共晶或前药,其中:R1选自苯环、噻吩环、呋喃环、吡咯环、噻唑环、噁唑环、咪唑环、异噻唑环、异噁唑环、吡唑环、吡啶环、嘧啶环、吡嗪环、喹啉环、异喹啉环、苯并异噻唑环、吲唑环、1H-吡唑[4,3-b]吡啶环或所述的苯环、噻吩环、呋喃环、吡咯环、噻唑环、噁唑环、咪唑环、异噻唑环、异噁唑环、吡唑环、吡啶环、嘧啶环、吡嗪环、喹啉环、异喹啉环、苯并异噻唑环、吲唑环、1H-吡唑[4,3-b]吡啶环或任选进一步被0至5个R1a取代;R1a选自F、Cl、Br、硝基、氨基、氰基、羟基、羧基、-C(=O)OBn、-CF3、甲基、乙基、丙基、异丙基、丁基、异丁基、叔丁基、乙炔基、丙炔基、炔丙基、甲氧基、乙氧基、丙氧基、异丙氧基、羟甲基、羟乙基、-C(CH3)2OH、苯氧基、苯胺基、苯基、2-噻吩基、环丙基、环丁基、环戊基、环己基或-CH2-环丙基;W选自键或-C(=O)-;条件是W选自-C(=O)-时,R1被1至5个R1a取代,且至少有1个R1a选自乙炔基、丙炔基、炔丙基或-CH2-环丙基;R2、R5各自独立的选自亚甲基、亚乙基、亚丙基、-CH2CH(CH3)-、-CH(CH3)CH2-、-C(CH3)2CH2-、-CH2C(CH3)2-、亚丁基、-CH(CH3)CH2CH2-、-CH2CH(CH3)CH2-或-CH2CH(CH3)CH2-;R3、R4各自独立的选自亚甲基、亚乙基、亚丙基或亚丁基;X1和X2各自独立的选自键、-O-、-C(=O)-、-C(=O)O-、-OC(=O)-、-C(=O)NRx-、-NRxC(=O)-、-OC(=O)NRx-、-NRxC(=O)O-或-NRx-;Rx各自独立选自H、甲基、乙基、丙基、异丙基、丁基、异丁基、戊基、环丙基、环丁基、环戊基、环己基、-CH2-环丙基、-CH2-环丁基或-CH2-环戊基;作为选择R3-X1-R4-X2-R5可为亚乙基;R6、R7各自独立的选自H、甲基或乙基;
- 根据权利要求5所述的化合物或其立体异构体、水合物、代谢产物、溶剂化物、药学上可接受的盐、共晶或前药,其中:R1选自苯环、噻吩环、呋喃环、吡咯环、噻唑环、噁唑环、咪唑环、异噻唑环、异噁唑环、吡唑环、吡啶环、吡嗪环、嘧啶环、喹啉环、异喹啉环、苯并异噻唑环、吲唑环、1H-吡唑[4,3-b]吡啶环或所述的苯环、噻吩环、呋喃环、吡咯环、噻唑环、噁唑环、咪唑环、异噻唑环、异噁唑环、吡唑环、吡啶环、嘧啶环、吡嗪环、喹啉环、异喹啉环、苯并异噻唑环、吲唑环、1H-吡唑[4,3-b]吡啶环或任选进一步被0至5个R1a取代;R1a选自F、Cl、Br、硝基、氨基、氰基、羟基、羧基、-C(=O)OBn、-CF3、甲基、乙基、丙基、异丙基、丁基、异丁基、叔丁基、乙炔基、丙炔基、炔丙基、甲氧基、乙氧基、丙氧基、异丙氧基、羟甲基、羟乙基、-C(CH3)2OH、苯氧基、苯胺基、苯基、2-噻吩基、环丙基、环丁基、环戊基、环己基或-CH2-环丙基;Ra选自F、Cl、Br、I、甲基、乙基、甲氧基、乙氧基或环丙烷基;条件是W选自-C(=O)-时,R1被1至5个R1a取代,且至少有1个R1a选自乙炔基、丙炔基、炔丙基、或-CH2-环丙基;Rx选自H、甲基、乙基、丙基、异丙基、丁基、异丁基、戊基、环丙基、环丁基、环戊基、环己基、-CH2-环丙基、-CH2-环丁基或-CH2-环戊基;
- 一种药物组合物,所述的药物组合物含有治疗有效剂量的根据利要求1~8中任一项所述的化合物或其立体异构体、水合物、代谢产物、溶剂化物、药学上可接受的盐、共晶或前药,以及药学上可接受的载体、稀释剂、佐剂、媒介物或赋形剂;所述的组合物还可进一步包括一种或多种其他治疗剂;优选的,所述其他治疗剂选自PDE4抑制剂、M受体拮抗剂、皮质类固醇和β-肾上腺素受体激动剂中的一种或多种。
- 权利要求1~8中任一项所述的化合物或其立体异构体、水合物、代谢产物、溶剂化物、药学上可接受的盐、共晶或前药,或权利要求9所述的药物组合物在制备用于治疗气道阻塞性疾病的药物中的应用,所述的气道阻塞性疾病优选哮喘、慢性阻塞性肺疾病或支气管炎。
- 一种治疗气道阻塞性疾病的方法,所述方法包括给药权利要求1~8中任一项所述的化合物或其立体异构体、水合物、代谢产物、溶剂化物、药学上可接受的盐、共晶或前药,或权利要求9所述的药物组合物,所述的气道阻塞性疾病优选哮喘、慢性阻塞性肺疾病或支气管炎。
- 一种制备式(I)所示化合物的中间体,该中间体选自式(M-1)、式(M-2)或式(M-3)所示化合物或者其立体异构体和药学上可接受的盐:其中:R1、W、R2、A、R3、X1、R4的定义如权利要求1~8中任一项所述;Rm1选自氨基保护基或H;Rm2选自-COOH、-COOC1-6烷基、-X2-R5-OH、-X2-R5-离去基团、-X2-Rm3CHO、-X2-Rm3C(OC1-6烷基)2或所述烷基任选进一步被0至4个F、Cl、Br或C1-4烷基所取代;X2的定义如权利要求1~8中任一项所述;Rm3选自较R5少一个碳原子的亚烷基,R5的定义如权利要求1~8中任一项所述;离去基团选自氯、溴、碘、-OMs或OTs;n1选自1、2或3;
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN201780033350.XA CN109195975B (zh) | 2016-09-30 | 2017-09-29 | 一种二氮杂螺[5.5]十一碳烷衍生物及其用途 |
Applications Claiming Priority (2)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN201610867511.2 | 2016-09-30 | ||
CN201610867511 | 2016-09-30 |
Publications (1)
Publication Number | Publication Date |
---|---|
WO2018059537A1 true WO2018059537A1 (zh) | 2018-04-05 |
Family
ID=61763293
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
PCT/CN2017/104286 WO2018059537A1 (zh) | 2016-09-30 | 2017-09-29 | 一种二氮杂螺[5.5]十一碳烷衍生物及其用途 |
Country Status (2)
Country | Link |
---|---|
CN (1) | CN109195975B (zh) |
WO (1) | WO2018059537A1 (zh) |
Citations (6)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2011012897A1 (en) * | 2009-07-31 | 2011-02-03 | Astrazeneca Ab | New combinations for the treatment of asthma |
CN102083839A (zh) * | 2008-02-06 | 2011-06-01 | 阿斯利康(瑞典)有限公司 | 化合物 |
WO2012085582A1 (en) * | 2010-12-23 | 2012-06-28 | Astrazeneca Ab | Compound |
WO2012085583A1 (en) * | 2010-12-23 | 2012-06-28 | Astrazeneca Ab | New compound |
CN102625808A (zh) * | 2009-07-31 | 2012-08-01 | 阿斯利康(瑞典)有限公司 | 螺环酰胺衍生物 |
CN103249418A (zh) * | 2010-10-07 | 2013-08-14 | 阿斯利康(瑞典)有限公司 | 新的组合 |
-
2017
- 2017-09-29 CN CN201780033350.XA patent/CN109195975B/zh not_active Expired - Fee Related
- 2017-09-29 WO PCT/CN2017/104286 patent/WO2018059537A1/zh active Application Filing
Patent Citations (6)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN102083839A (zh) * | 2008-02-06 | 2011-06-01 | 阿斯利康(瑞典)有限公司 | 化合物 |
WO2011012897A1 (en) * | 2009-07-31 | 2011-02-03 | Astrazeneca Ab | New combinations for the treatment of asthma |
CN102625808A (zh) * | 2009-07-31 | 2012-08-01 | 阿斯利康(瑞典)有限公司 | 螺环酰胺衍生物 |
CN103249418A (zh) * | 2010-10-07 | 2013-08-14 | 阿斯利康(瑞典)有限公司 | 新的组合 |
WO2012085582A1 (en) * | 2010-12-23 | 2012-06-28 | Astrazeneca Ab | Compound |
WO2012085583A1 (en) * | 2010-12-23 | 2012-06-28 | Astrazeneca Ab | New compound |
Also Published As
Publication number | Publication date |
---|---|
CN109195975A (zh) | 2019-01-11 |
CN109195975B (zh) | 2022-01-04 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
JP6525437B2 (ja) | 抗線維性ピリジノン | |
US10479797B2 (en) | Modulators of the beta-3 adrenergic receptor useful for the treatment or prevention of disorders related thereto | |
AU2011208139B2 (en) | Piperazine compound having a PGDS inhibitory effect | |
WO2019205983A1 (zh) | 氧杂螺环类化合物及其制备方法和用途 | |
WO2014146493A1 (en) | Acyclic cyanoethylpyrazolo pyridones as janus kinase inhibitors | |
TWI665197B (zh) | 具有β2受體激動及M3受體拮抗活性的苯並環衍生物及其在醫藥上的用途 | |
TW201730150A (zh) | 氮雜環醯胺衍生物、其製備方法及醫藥用途 | |
KR20220141331A (ko) | P2x3 조정제 | |
CN106565674B (zh) | 一种八氢环戊烷并[c]吡咯衍生物及其制备方法和在医药上的用途 | |
WO2016180349A1 (zh) | 具有β2受体激动及M受体拮抗活性的联苯衍生物及其在医药上的用途 | |
CN107074816A (zh) | 一种杂环衍生物及其制备方法和在医药上的用途 | |
CN107531721B (zh) | 用于调节法尼醇x受体的稠合的三环吡唑衍生物 | |
WO2018059537A1 (zh) | 一种二氮杂螺[5.5]十一碳烷衍生物及其用途 | |
WO2021228217A1 (zh) | 可用作RORγ调节剂的苯胺类化合物 | |
CN110590776A (zh) | 一种联苯衍生物及其制备方法和在医药上的用途 | |
CN106336406B (zh) | 具有β2受体激动及M受体拮抗活性的八氢并环戊二烯衍生物及其在医药上的用途 | |
CN107849014B (zh) | 一种联苯衍生物及其制备方法和在医药上的用途 | |
WO2022012534A1 (zh) | 含氮杂环化合物、药物组合物和应用 | |
CN107849035B (zh) | 一种苯基杂环衍生物及其在医药上的用途 | |
WO2022184103A1 (zh) | 三并环化合物及其药物组合物和应用 | |
WO2023208165A1 (zh) | 一种含氮杂环衍生物及其组合物和药学上的应用 | |
WO2022166980A1 (zh) | 杂芳基并哌啶类衍生物及其药物组合物和应用 | |
TW202310830A (zh) | 用於治療疾病之β-內醯胺衍生物 |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
121 | Ep: the epo has been informed by wipo that ep was designated in this application |
Ref document number: 17854987 Country of ref document: EP Kind code of ref document: A1 |
|
NENP | Non-entry into the national phase |
Ref country code: DE |
|
122 | Ep: pct application non-entry in european phase |
Ref document number: 17854987 Country of ref document: EP Kind code of ref document: A1 |