WO2021175196A1 - 一种吡咯烷类整合素调节剂及其用途 - Google Patents

一种吡咯烷类整合素调节剂及其用途 Download PDF

Info

Publication number
WO2021175196A1
WO2021175196A1 PCT/CN2021/078537 CN2021078537W WO2021175196A1 WO 2021175196 A1 WO2021175196 A1 WO 2021175196A1 CN 2021078537 W CN2021078537 W CN 2021078537W WO 2021175196 A1 WO2021175196 A1 WO 2021175196A1
Authority
WO
WIPO (PCT)
Prior art keywords
compound
formula
methyl
esi
butyl
Prior art date
Application number
PCT/CN2021/078537
Other languages
English (en)
French (fr)
Inventor
李小林
谢永华
石峰
李子忠
吴奕钦
马晓芸
张士猛
吴钎
Original Assignee
清药同创(北京)药物研发中心有限公司
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by 清药同创(北京)药物研发中心有限公司 filed Critical 清药同创(北京)药物研发中心有限公司
Publication of WO2021175196A1 publication Critical patent/WO2021175196A1/zh

Links

Classifications

    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D471/00—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, at least one ring being a six-membered ring with one nitrogen atom, not provided for by groups C07D451/00 - C07D463/00
    • C07D471/02—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, at least one ring being a six-membered ring with one nitrogen atom, not provided for by groups C07D451/00 - C07D463/00 in which the condensed system contains two hetero rings
    • C07D471/04—Ortho-condensed systems
    • 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
    • A61K31/435—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom
    • A61K31/4353—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom ortho- or peri-condensed with heterocyclic ring systems
    • A61K31/4375—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom ortho- or peri-condensed with heterocyclic ring systems the heterocyclic ring system containing a six-membered ring having nitrogen as a ring heteroatom, e.g. quinolizines, naphthyridines, berberine, vincamine
    • 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
    • A61K31/435—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom
    • A61K31/44—Non condensed pyridines; Hydrogenated derivatives thereof
    • A61K31/4427—Non condensed pyridines; Hydrogenated derivatives thereof containing further heterocyclic ring systems
    • A61K31/444—Non condensed pyridines; Hydrogenated derivatives thereof containing further heterocyclic ring systems containing a six-membered ring with nitrogen as a ring heteroatom, e.g. amrinone
    • 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
    • A61K31/435—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom
    • A61K31/47—Quinolines; Isoquinolines
    • A61K31/4709—Non-condensed quinolines and containing further heterocyclic rings
    • 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
    • A61K31/495—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with two or more nitrogen atoms as the only ring heteroatoms, e.g. piperazine or tetrazines
    • A61K31/505—Pyrimidines; Hydrogenated pyrimidines, e.g. trimethoprim
    • A61K31/506—Pyrimidines; Hydrogenated pyrimidines, e.g. trimethoprim not condensed and containing further heterocyclic rings
    • 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
    • A61K31/5375—1,4-Oxazines, e.g. morpholine
    • A61K31/5377—1,4-Oxazines, e.g. morpholine not condensed and containing further heterocyclic rings, e.g. timolol
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
    • A61P1/16—Drugs for disorders of the alimentary tract or the digestive system for liver or gallbladder disorders, e.g. hepatoprotective agents, cholagogues, litholytics
    • 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
    • A61P13/00—Drugs for disorders of the urinary system
    • A61P13/10—Drugs for disorders of the urinary system of the bladder
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P13/00—Drugs for disorders of the urinary system
    • A61P13/12—Drugs for disorders of the urinary system of the kidneys
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P17/00—Drugs for dermatological disorders
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P17/00—Drugs for dermatological disorders
    • A61P17/06—Antipsoriatics
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P19/00—Drugs for skeletal disorders
    • A61P19/08—Drugs for skeletal disorders for bone diseases, e.g. rachitism, Paget's disease
    • A61P19/10—Drugs for skeletal disorders for bone diseases, e.g. rachitism, Paget's disease for osteoporosis
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P25/00—Drugs for disorders of the nervous system
    • A61P25/02—Drugs for disorders of the nervous system for peripheral neuropathies
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P25/00—Drugs for disorders of the nervous system
    • A61P25/04—Centrally acting analgesics, e.g. opioids
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P27/00—Drugs for disorders of the senses
    • A61P27/02—Ophthalmic agents
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P27/00—Drugs for disorders of the senses
    • A61P27/02—Ophthalmic agents
    • A61P27/06—Antiglaucoma agents or miotics
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P29/00—Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P3/00—Drugs for disorders of the metabolism
    • A61P3/08—Drugs for disorders of the metabolism for glucose homeostasis
    • A61P3/10—Drugs for disorders of the metabolism for glucose homeostasis for hyperglycaemia, e.g. antidiabetics
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • A61P35/02—Antineoplastic agents specific for leukemia
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P9/00—Drugs for disorders of the cardiovascular system
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P9/00—Drugs for disorders of the cardiovascular system
    • A61P9/04—Inotropic agents, i.e. stimulants of cardiac contraction; Drugs for heart failure
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P9/00—Drugs for disorders of the cardiovascular system
    • A61P9/10—Drugs for disorders of the cardiovascular system for treating ischaemic or atherosclerotic diseases, e.g. antianginal drugs, coronary vasodilators, drugs for myocardial infarction, retinopathy, cerebrovascula insufficiency, renal arteriosclerosis
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D401/00—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom
    • C07D401/02—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom containing two hetero rings
    • C07D401/12—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom containing two hetero rings linked by a chain containing hetero atoms as chain links
    • 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 invention belongs to the field of pharmaceutical compounds, and specifically relates to a pyrrolidine integrin regulator and its use.
  • the integrin family is a type of widely distributed transmembrane glycoproteins that connect the extracellular matrix environment with intracellular signal transduction. Integrins consist of ⁇ subunits and ⁇ subunits connected by non-covalent bonds to form heterodimers. At present, 18 ⁇ subunits and 8 ⁇ subunits have been found, which can be combined into at least 24 integrin dimers.
  • the integrin family of cell adhesion molecules is the main connection material between extracellular matrix, inflammatory cells, fibroblasts and parenchymal cells, and is closely related to the occurrence, maintenance and development of tissue fibrosis. The process mediates many key cell-cell and cell-extracellular matrix interactions. (Manninen et al,Proteomics,2017,17(3-4):1600022.).
  • the ⁇ V family is mainly involved in the fibrosis process of body tissues.
  • the ⁇ V family of integrins includes 5 subtypes ( ⁇ V ⁇ 1, ⁇ V ⁇ 3, ⁇ V ⁇ 5, ⁇ V ⁇ 6, and ⁇ V ⁇ 8), which are lowly expressed in normal cells of various tissues, and highly expressed in fibrous tissue cells.
  • TGF- ⁇ is an important factor involved in the formation of tissue fibrosis, especially TGF- ⁇ 1. In inflammation and fibrosis, TGF- ⁇ 1 is involved in physiological repair and collagen accumulation.
  • Integrin ⁇ V family proteins participate in the activation of latent TGF- ⁇ molecules, and induce excessive autoimmune and inflammatory responses by activating TGF- ⁇ , and promote tissue fibrosis.
  • Integrin ⁇ v ⁇ 6 and ⁇ v ⁇ 1 play a major role in renal and lung fibrosis.
  • ⁇ v ⁇ 3/5 is more common in cardiac fibrosis, while ⁇ v ⁇ 1 accounts for liver fibrosis.
  • Integrin ⁇ v ⁇ 1 is a low-affinity fibronectin receptor, which is highly expressed in basal epithelial cells and can promote the migration of keratinocytes on the underlying fibronectin EDA. Blocking the interaction of integrin ⁇ v ⁇ 1 and TGF- ⁇ 1 helps to inhibit the activity of TGF- ⁇ 1 and block the process of fibrosis.
  • integrin ⁇ v ⁇ 1 can also participate in the synthesis of gingival fibroblasts by activating latent TGF ⁇ 1 (Jakhu et al, Journal of oral biology and craniofacial research, 2018, 8(2): 122.).
  • TGF ⁇ 1 latent TGF ⁇ 1
  • integrin ⁇ v ⁇ 6 the research on ⁇ v ⁇ 6 is more in-depth. It has been reported that the expression of integrin ⁇ v ⁇ 6 is very low in normal lung tissue, but when inflammation and fibrosis occur in lung injury, ⁇ v ⁇ 6 is rapidly expressed (Hatley et al, Angewandte Chemie International Edition, 2018, 57(13): 3298.).
  • integrin ⁇ V ⁇ 6 In patients with primary biliary cirrhosis (PBC), alcoholic fatty liver, hepatitis B, hepatitis C and other diseases, the mRNA expression of integrin ⁇ V ⁇ 6 is increased.
  • the expression of integrin ⁇ V ⁇ 6 in chronic inflammation and fibrotic diseases related to kidney disease is significantly higher than that of normal kidney tissue.
  • integrin ⁇ V ⁇ 6 is significantly highly expressed in biopsy samples of patients with diabetes, pulmonary hemorrhagic nephritis syndrome, Alport syndrome, and lupus nephritis (Koivisto et al, The international journal of biochemistry & cell biology, 2018, 99: 186.).
  • Tissue fibrosis can occur in a variety of organs. It is a relatively common fibrotic disease including idiopathic pulmonary fibrosis (IPF), non-alcoholic fatty liver (NASH), liver cirrhosis, renal fibrosis, scleroderma, Myocardial fibrosis and so on. Tissue damage and inflammation are important causes of fibrosis. Inflammation leads to necrosis of organ parenchymal cells, local immune cells are activated, and a variety of blood cells enter the injury site. The activated immune cells produce a large number of highly biologically active cytokines and chemokines, leading to the local activation of mesenchymal cells.
  • ECM extracellular matrix
  • chemokines chemokines
  • angiogenic factors angiogenic factors
  • pathological changes and tissue fibrosis Rost-Blum et al, Matrix Biology, 2018, 68: 122.
  • the main feature of fibrosis is the formation and deposition of excessive fibrous connective tissue. Chronic damage with fibrosis will destroy the tissue structure and cause organ dysfunction, eventually leading to organ failure.
  • the present invention provides a compound represented by formula I and its racemates, stereoisomers, tautomers, isotopic labels, nitrogen oxides, solvates, poly Crystal form, metabolite, ester, prodrug or pharmaceutically acceptable salt thereof:
  • the W is selected from -O-, -NR 2 -, -S-;
  • the R 2 and R 3 are each independently selected from H, C 1 -C 12 aliphatic hydrocarbon group, C 3-12 cycloalkyl, C 3-12 cycloalkyl-C 1 -C 12 aliphatic hydrocarbon group, -L 5- Ar, and at least one of R 2 and R 3 is selected from -L 5 -Ar;
  • the R 4 is independently selected from H, C 1 -C 12 aliphatic hydrocarbon group
  • L 3 is selected from the bond, -CH 2 -;
  • L 4 is selected from the bond,- CH 2 -;
  • L 5 is selected from the bond, -CH 2 -;
  • the "halogen” is selected from F, Cl, Br, and I;
  • the "aliphatic hydrocarbon group” is selected from alkyl, alkenyl, and alkynyl;
  • the R 1 and R 5 are each independently selected from halogen, C 1 -C 6 aliphatic hydrocarbon group, for example, selected from F, Cl, Br, I, methyl, ethyl, N-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, n-pentyl, isopentyl, neopentyl, n-hexyl.
  • the R 2 is selected from H, C 1 -C 6 aliphatic hydrocarbon group or C 3-8 cycloalkyl-C 1 -C 6 aliphatic hydrocarbon group
  • R 3 is selected from -L 5 -Ar or the R 2 is selected from -L 5 -Ar and R 3 is selected from H, C 1 -C 6 aliphatic hydrocarbon group or C 3-8 cycloalkyl-C 1 -C 6 aliphatic hydrocarbon group; more preferably,
  • One of R 2 or R 3 is selected from H, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, n-pentyl, isopentyl, neopentyl, N-hexyl, cyclopropylmethyl, cyclobutylmethyl, cyclopentylmethyl, cyclohexylmethyl.
  • the Ar is selected from the following groups optionally substituted by one, two or more R b : C 6 -C 14 aryl, 5-10 membered heterocyclic group, 5-6 Heteroaryl groups, preferably, Ar is selected from phenyl optionally substituted with one, two or more R b , naphthyl, 2,3-dihydrobenzofuranyl, benzofuranyl, benzofuranyl Pyranyl, 3,4-dihydro-2H-1-benzopyranyl (chromanyl), 2,3-dihydrobenzo[b][1,4]dioxanyl, pyridyl, Pyrimidyl, indazolyl (1H-indazolyl, 2H-indazolyl), indolyl, isoindolyl, quinolinyl, isoquinolinyl, quinazolinyl, benzoxazolyl;
  • the R b is selected from H, halogen, CN, or the following groups optionally substituted by one, two or more R c : C 1 -C 6 aliphatic hydrocarbon group, C 1 -C 6 aliphatic hydrocarbon group oxy, C 1 -C 6 aliphatic hydrocarbon group -SO 2 -, C 1 -C 6 aliphatic hydrocarbon group -NH-, two (C 1 -C 6 aliphatic hydrocarbon group) N-, C 6-10 Aryl, C 6-10 aryloxy, C 6-10 aryl-SO 2 -, C 6-10 aryl-NH-, 5-6 membered heteroaryl, 5-6 membered heteroaryloxy , 5-6 membered heteroaryl-SO 2 -, 5-6 membered heteroaryl-NH-, 5-6 membered heterocyclyl, 5-6 membered heterocyclyloxy, 5-6 membered heterocyclyl- SO 2 -, 5-6 member
  • the R 4 is independently selected from H, C 1 -C 6 aliphatic hydrocarbon groups, preferably, selected from methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl Base, tert-butyl, n-pentyl, isopentyl, neopentyl, n-hexyl;
  • the Ar is selected from the following groups:
  • L 5 (not a bond), R b , and R c are selected from the definitions described in Formula I.
  • one of R 2 or R 3 is selected from the following groups:
  • configuration at *S 1 can be selected from:
  • the configuration at *S 2 can be selected from:
  • the formula I may be selected from a specific stereo configuration including one chiral center, or a compound structure of a combination of two or more chiral centers with different specific stereo configurations.
  • formula II the structure of formula I may be selected from formula II:
  • R 1 , R 2 , R 3 , R 4 , R 5 , L 1 , L 2 , L 3 , L 4 and other chiral centers are as defined in Formula I above.
  • the structure of the formula II is further represented by the following formula IIa-IIt:
  • the compound of formula I is further preferably the following formula III (formula IIIa, formula IIIb), formula IV (formula IVa, formula IVb):
  • the compound of formula I is further preferably the following formula V to formula XVII:
  • R 1 , R 2 , R 3 , R 4 , R 5 , n, m and other chiral centers are as defined in Formula I above.
  • the compound represented by formula I (including formula II to formula XVII) and its racemates, stereoisomers, tautomers, isotope labels, nitrogen oxides, and solvates Among the polymorphs, metabolites, esters, prodrugs, or pharmaceutically acceptable salts thereof, illustrative, non-limiting specific examples of the compound of formula I are as follows:
  • the mark with an asterisk indicates that the structure has chirality at the carbon position indicated by the asterisk, so it can exist in the form of a mixture of isomers with opposite conformations at the marked position, and can be used by hand
  • Two isomers with the described structure can be obtained by sexual resolution (see Example 3, for example, the epimer mixture form of compound 5 can be obtained by chiral resolution to obtain compounds 5a and 5b with different asterisk positions and configurations) .
  • the stereoisomers of the compound of formula I may be further selected from, for example, the following structures:
  • the present invention also provides the compounds represented by formula I (including formula II to formula XVII) and their racemates, stereoisomers, tautomers, isotopic labels, nitrogen oxides, solvates, and polymorphs
  • the compounds of the present invention can be synthesized by combining the following methods with synthetic methods known in the field of synthetic organic chemistry or related modification methods recognized by those skilled in the art. Those skilled in the art know that according to the specific target structure, one or more of the following schemes can be optionally used to combine, or any step of one or more of the schemes can be combined to obtain a synthetic scheme.
  • the preparation method of the compound of the present invention includes: reacting a raw material containing a naphthyridine ring structure with a raw material containing a pyrrolidine structure in a suitable reagent under suitable conditions, and optionally, under suitable conditions, Carry out protective group, deprotection, substitution, condensation, reductive amination or hydrolysis steps. Specifically, it can be synthesized with reference to the following scheme.
  • the preparation of the compound of the present invention includes one or more of the following steps:
  • the R 1 , R 3 , R 4 (R 4 ⁇ H), R 5 , L 1 , L 2 , L 3 , L 4 are as defined in the aforementioned formula I;
  • the Lx is selected from L 1 -X 1 , where X 1 is a leaving group, selected from, for example, OTs, OMs, OTf, halogen (Cl, Br, I), or the Lx is selected from the L 1 group (when there is a terminal -CH 2-
  • the terminal -CH 2 - is replaced by a group of CHO or COOH, for example, selected from -(CH 2 ) m-1 -CHO or -(CH 2 ) m-1 -COOH, the m is as described in the aforementioned formula I Definition;
  • the PG 1 is a protecting group on N, selected from Boc- etc.;
  • compound A-3 is synthesized by using compounds A-1 and A-2, and the reaction conditions can be selected from (i) NaBH(OAc) 3
  • R 2 , R 3 , R 4 , and m are as defined in the aforementioned formula I, wherein m is not 0; the X 1 and PG 1 are as defined above;
  • the preparation of the compound of the present invention further includes one or more of the following steps;
  • the R 3 , R 4 , R 5 , L 4 are as defined in the aforementioned formula I; the PG 1 is as defined before; the compounds B-1 and B-2 are used in reducing agents, acids, organic Compound B-3 is obtained in the presence of a solvent, the reducing agent is selected from NaBH 3 CN, the acid is selected from acetic acid, and the organic solvent is selected from alcohol reagents such as methanol and ethanol; the compound B-3 and the compound B-5 is hydrolyzed in an organic solvent under acidic conditions to obtain B-4 and B-6, the acidic conditions are selected from TFA, and the organic solvent is selected from DCM; the B-3 is obtained by NH methylation reaction For compound B-5, the methylation reaction conditions are paraformaldehyde, NaBH(OAC) 3 , NaBH 3 CN, and carried out in an organic solvent, and the organic solvent is selected from alcohol reagents, such as methanol and ethanol.
  • the preparation of the compound of the present invention includes the following steps:
  • compound C-1 can be resolved into compounds C-2 and C-3, and the adopted resolution conditions include the use of a chiral chromatography column.
  • the R 1 , R 2 , R 3 , R 4 , R 5 , L 1 , and L 3 are as defined in the aforementioned formula I.
  • the preparation of the compound of the present invention includes one or more of the following steps;
  • X 2 is a leaving group, selected from halogens, such as Cl, Br, I; R 1 , R 2 , R 3 , R 4 , R 5 are as defined in the aforementioned formula I, and the PG 1 Same as the aforementioned definition.
  • the compound D-1 is reacted in the presence of a basic reagent, a catalyst and an organic reagent to obtain compound D-2
  • the basic reagent is selected from alkali metal or alkaline earth metal carbonates, such as K 2 CO 3 , Na 2 CO 3
  • the catalyst is an iodide, such as NaI
  • the organic reagent is selected from acetonitrile, DMF, and DMSO; the compound D-2 undergoes NH methylation to obtain the compound D-3, the methyl group
  • the chemical reaction conditions are paraformaldehyde, NaBH(OAC) 3 , NaBH 3 CN, and an organic solvent.
  • the organic solvent is selected from alcohol reagents, such as methanol and ethanol; the compound D-4 is in H 2 SO 4 , Compound D-5 is obtained in the presence of an alkyl alcohol reagent, and the alkyl alcohol reagent can be selected from methanol; the compound D-5 is reacted in the presence of a halogenated reagent, an organic solvent, and an initiator to obtain compound D-1.
  • alcohol reagents such as methanol and ethanol
  • the compound D-4 is in H 2 SO 4
  • Compound D-5 is obtained in the presence of an alkyl alcohol reagent, and the alkyl alcohol reagent can be selected from methanol; the compound D-5 is reacted in the presence of a halogenated reagent, an organic solvent, and an initiator to obtain compound D-1.
  • the halogenated reagent is selected from NBS, CBr 4 , the initiator is selected from AIBN, and the organic solvent is selected from carbon tetrachloride, dichloromethane, chloroform, 1,4-dioxane, and tetrahydrofuran;
  • the D-1 and D-6 produce D-7 in the presence of an alkaline reagent and an organic solvent, the alkaline reagent is selected from alkali metal or alkaline earth metal carbonates, and the organic solvent is selected from dichloromethane, Trichloromethane, carbon tetrachloride.
  • the preparation of the compound of the present invention includes one or more of the following steps:
  • the R 2 , R 4 , and R 5 are as defined in the aforementioned formula I, and the R 2 is selected from -L 5 -Ar; the PG 1 is the same as the aforementioned definition; the compound E-1 and The organic amine reagent reacts to obtain E-2, the reaction is carried out in the presence of a reducing reagent, an acidic reagent, and an organic solvent, the reducing reagent is selected from BH 3 ⁇ THF, the acidic reagent is selected from acetic acid, and the organic solvent is selected From THF; The compound E-2 reacts with E-3 to obtain E-4, and the reaction is carried out in the presence of a basic reagent, a catalyst, and an organic solvent.
  • the basic reagent is selected from the group consisting of alkali metal or alkaline earth metal carbon
  • the acid salt is, for example, K 2 CO 3 , Na 2 CO 3
  • the catalyst is iodide, for example, NaI
  • the organic reagent is selected from acetonitrile, DMF, and DMSO.
  • the preparation of the compound of the present invention includes the following steps:
  • the p, q, R b , R 4 , R 5 are as defined in the aforementioned formula I; the PG 1 is the same as the aforementioned definition; the compound F-1 is reacted with F-2 to obtain F-3,
  • the reaction is carried out in the presence of a reducing reagent, an acidic reagent, and an organic solvent, the reducing reagent is selected from sodium triacetoxyborohydride, sodium cyanoborohydride, the acidic reagent is selected from acetic acid, and the organic solvent is selected Self-alcoholic reagents, such as methanol, ethanol.
  • the preparation of the compound of the present invention includes one or more of the following steps:
  • the R 4 is as defined in the aforementioned formula I, and the X 3 is selected from halogens, such as F, Cl, Br, I; the compound G-1 is reacted with a halogenated alkane reagent to produce the compound G-2,
  • the reaction is carried out under the conditions of an alkaline reagent and an organic solvent.
  • the alkaline reagent is selected from alkali metal or alkaline earth metal carbonates, such as K 2 CO 3 , Na 2 CO 3 , and the organic reagent is selected from acetonitrile and DMF.
  • the compound G-2 reacts with G-3 to produce G-4, and the reaction is carried out under the conditions of an alkaline reagent and an organic solvent, the alkaline reagent is selected from NaH, LiH, KH, and the organic solvent is selected from Toluene, benzene.
  • the preparation of the compound of the present invention includes the following steps:
  • the R b is as defined in the aforementioned formula I
  • the compound H-1 (hydrochloride) is reacted to obtain H-2 and H-3
  • the reaction is carried out in the presence of an acidic reagent, water, and an organic solvent
  • the acidic reagent is selected from hydrochloric acid
  • the organic solvent is selected from alcohol reagents, such as methanol and ethanol.
  • the reaction is preferably carried out under microwave conditions.
  • the preparation of the compound of the present invention includes one or more of the following steps:
  • the R 4 and R b are as defined in the aforementioned formula I, and the X 4 is selected from halogens, such as F, Cl, Br, I; the compound I-1 is reacted to form I-2, and the reaction is
  • the organic solvent, Pb(OAc) 2 is carried out in the presence of an alkaline reagent, the alkaline reagent is selected from triethylamine, pyridine, etc., and the organic solvent is selected from 1,4-dioxane.
  • the I-2 generates I-3 in the presence of an alkaline agent, and the alkaline agent is selected from NaH, KH, and LiH.
  • the preparation of the compound of the present invention includes the following steps:
  • the R 4 and R b are as defined in the aforementioned formula I, the compound J-1 and J-2 react to form J-3, and the reaction is carried out in the presence of thionyl chloride, magnesium chloride, and an organic solvent.
  • the organic solvent is selected from tetrahydrofuran, dichloromethane, and chloroform.
  • the preparation of the compound of the present invention includes one or more of the following steps:
  • the preparation of the compound of the present invention includes the following steps:
  • R 1 , m are as defined in the aforementioned formula I, and the PG 1 is the same as the aforementioned definition; the compound L-1 is hydrolyzed to L-2, and the reaction is carried out under alkaline reagent, organic solvent/water reaction conditions
  • the alkaline reagent is selected from alkali metal or alkaline earth metal hydroxides, for example, NaOH, KOH, LiOH, and the organic solvent is selected from alcohol reagents, for example, methanol, ethanol; the compound L-3
  • the reaction obtains L-4, which is reacted in the presence of a reducing reagent and an organic solvent, the reducing reagent is selected from DIBAL-H, and the organic solvent is selected from THF, dichloromethane, chloroform and the like.
  • the preparation of the compound of the present invention includes one or more of the following steps:
  • R b , R 3 , R 4 , and R 5 are as defined in the aforementioned formula I, and the PG 1 is the same as the aforementioned definition.
  • the preparation of the compound of the present invention includes the following steps:
  • the preparation of the compound of the present invention includes one or more of the following steps:
  • the R 4 and R 5 are as defined in the aforementioned formula I
  • the PG 1 is the same as the aforementioned definition
  • the X 5 is selected from halogen, such as F, Cl, Br, I.
  • the preparation of the compound of the present invention includes one or more of the following steps:
  • R 4 is defined as the aforementioned formula I
  • the PG 1 is the same as the aforementioned definition
  • the X 6 is selected from halogens, such as F, Cl, Br, I.
  • the preparation of the compound of the present invention includes one or more of the following steps:
  • R b is selected from C 1 -C 12 alkoxy, C 3 -C 8 cycloalkyloxy or 3-12 membered heterocyclyloxy as defined in the previous formula I.
  • the compound P-1 obtains the compound P-2 in the presence of an alkyl alcohol and an acidic reagent, the alkyl alcohol is selected from methanol, ethanol, and the acidic reagent is selected from sulfuric acid; the compound P-2 is in the presence of a halogenated aliphatic hydrocarbon Or halogenated alicyclic hydrocarbon reagent, organic solvent, and alkaline reagent are reacted to obtain compound P-3.
  • the halogenated aliphatic hydrocarbon or halogenated alicyclic hydrocarbon reagent can be selected from halogenated (C 1 -C 12 )alkanes , Halogenated (C 3 -C 8 ) cycloalkanes or halogenated 3-12 membered heterocycles, for example selected from 2-iodopropane, 3-iodooxetane; the basic reagent is selected from alkali metals or Alkaline earth metal carbonates, such as potassium carbonate, sodium carbonate, cesium carbonate, and the organic solvent is selected from DMF, DMSO, and acetonitrile.
  • the preparation of the compound of the present invention includes one or more of the following steps:
  • X 7 is halogen, selected from F, Cl, Br, and I.
  • the preparation of the compound of the present invention includes the following steps:
  • R 4 is as defined in the aforementioned formula I
  • X 8 is halogen, selected from F, Cl, Br, and I.
  • the preparation of the compound of the present invention includes the following steps:
  • R b is as defined in Formula I above.
  • the preparation of the compound of the present invention includes the following steps:
  • R b is as defined in the aforementioned formula I
  • X 9 is halogen, selected from F, Cl, Br, I
  • the reaction conditions are in the presence of alkaline reagents, organic solvents and alkyl alcohol reagents.
  • the alkaline reagent is selected from alkali metal or alkaline earth metal hydroxides, such as KOH, NaOH
  • the organic solvent is selected from DMSO, DMF, acetonitrile
  • the alkyl alcohol reagent is selected from methanol, ethanol, and the like.
  • the preparation of the compound of the present invention includes the following steps:
  • R 4 is as defined in the aforementioned formula I
  • X 10 is halogen, selected from F, Cl, Br, and I.
  • the present invention further provides a pharmaceutical composition
  • a pharmaceutical composition comprising the compound of formula I of the present invention and its racemates, stereoisomers, tautomers, isotope markers, nitrogen oxides, solvates, and polymorphs. Forms, metabolites, esters, prodrugs or pharmaceutically acceptable salts thereof.
  • the pharmaceutical composition of the present invention further comprises a therapeutically effective amount of the compound of formula I of the present invention and its racemates, stereoisomers, tautomers, isotope markers, nitroxides Substances, solvates, polymorphs, metabolites, esters, prodrugs or pharmaceutically acceptable salts thereof and pharmaceutically acceptable carriers.
  • the carrier in the pharmaceutical composition is "acceptable", which is compatible with the active ingredient of the composition (and preferably capable of stabilizing the active ingredient) and is not harmful to the subject being treated.
  • One or more solubilizers can be used as pharmaceutical excipients for the delivery of active compounds.
  • the present invention further provides the compound of formula I and its racemates, stereoisomers, tautomers, isotope markers, nitrogen oxides, solvates, polymorphs, metabolites, esters, prodrugs or The application of the pharmaceutically acceptable salt or the pharmaceutical composition in the preparation of an integrin modulator.
  • the present invention further provides the compound of formula I and its racemates, stereoisomers, tautomers, isotope markers, nitrogen oxides, solvates, polymorphs, metabolites, esters, prodrugs or Use of the pharmaceutically acceptable salt or the pharmaceutical composition in the preparation of a medicament for preventing, regulating or treating diseases or disorders related to integrin activity.
  • the present invention further provides the compound of formula I and its racemates, stereoisomers, tautomers, isotope markers, nitrogen oxides, solvates, polymorphs, metabolites, esters, prodrugs or Use of the pharmaceutically acceptable salt or the pharmaceutical composition in the preparation of a medicament for treating fibrotic diseases, inflammatory diseases or cell proliferative diseases.
  • the present invention further provides the compound of formula I and its racemates, stereoisomers, tautomers, isotope markers, nitrogen oxides, solvates, polymorphs, metabolites, esters, prodrugs or Use of the pharmaceutically acceptable salt or the pharmaceutical composition in the preparation of a medicament for inhibiting the activation of TGF- ⁇ in cells.
  • the present invention also provides a method for modulating the activity of at least one integrin in a subject, the method comprising administering the compound of the present invention as a therapeutic agent.
  • the compound's modulating effect on integrin is to any one of ⁇ V ⁇ 1, ⁇ V ⁇ 3, ⁇ V ⁇ 5, ⁇ V ⁇ 6, and ⁇ V ⁇ 8, ⁇ 5 ⁇ 1, ⁇ 8 ⁇ 1 or ⁇ V ⁇ 1, ⁇ V ⁇ 3, ⁇ V ⁇ 5, ⁇ V ⁇ 6, and ⁇ V ⁇ 8, ⁇ 5 ⁇ 1, ⁇ 8 ⁇ 1 Or a combination of multiple adjustments.
  • the modulating effect is manifested as an inhibitory effect.
  • the inhibitory effect may be an inhibitory effect on ⁇ V ⁇ 1, ⁇ V ⁇ 3, ⁇ V ⁇ 5, ⁇ V ⁇ 6, and ⁇ V ⁇ 8, ⁇ 5 ⁇ 1, and ⁇ 8 ⁇ 1; in other embodiments, the inhibitory effect may be on ⁇ V ⁇ 1, ⁇ V ⁇ 3, ⁇ V ⁇ 5, One of ⁇ V ⁇ 6, and ⁇ V ⁇ 8, ⁇ 5 ⁇ 1, and ⁇ 8 ⁇ 1; in other embodiments, the inhibitory effect includes an inhibitory effect on ⁇ 8 ⁇ 1 and ⁇ V ⁇ 1; in other embodiments, the inhibitory effect includes an inhibitory effect on ⁇ 8 ⁇ 1 and ⁇ 5 ⁇ 1.
  • the inhibitory effect includes an inhibitory effect on ⁇ v ⁇ 3 and ⁇ v ⁇ 5; in some embodiments, the inhibitory effect includes an inhibitory effect on ⁇ 8 ⁇ 1, ⁇ v ⁇ 3, and ⁇ v ⁇ 5; in some embodiments, The inhibitory effect includes an inhibitory effect on ⁇ 8 ⁇ 1, ⁇ v ⁇ 1, and ⁇ 5 ⁇ 1; in some embodiments, the inhibitory effect includes an inhibitory effect on ⁇ 8 ⁇ 1, ⁇ v ⁇ 1, ⁇ v ⁇ 3, and ⁇ v ⁇ 5.
  • the integrin includes a combination of one or more of ⁇ V ⁇ 1, ⁇ V ⁇ 3, ⁇ V ⁇ 5, ⁇ V ⁇ 6, and ⁇ V ⁇ 8, ⁇ 5 ⁇ 1, and ⁇ 8 ⁇ 1.
  • the present invention also provides a method for treating a disease or condition, the method comprising administering to a patient in need of such treatment a therapeutically effective amount of at least one compound of the present invention alone, or optionally, in combination with another of the present invention.
  • a therapeutically effective amount of at least one compound of the present invention alone, or optionally, in combination with another of the present invention.
  • One compound and/or at least one other type of therapeutic agent combination is also provided.
  • the present invention also provides a method for inhibiting the activation of TGF- ⁇ in a cell, the method comprising administering a compound of formula I and its racemates, stereoisomers, tautomers, isotope markers, nitrogen oxides to the cell Compounds, solvates, polymorphs, metabolites, esters, prodrugs, or pharmaceutically acceptable salts thereof, or the pharmaceutical composition.
  • the disease or condition is associated with fibrosis, including lung, liver, kidney, heart, skin, eye, and pancreatic fibrosis.
  • the disease or disorder is related to a cell proliferative disorder such as cancer.
  • the cancer includes solid tumor growth or neoplasia.
  • the cancer includes tumor metastasis.
  • the cancer is bladder cancer, blood cancer, bone cancer, brain cancer, breast cancer, central nervous system cancer, cervical cancer, colon cancer, endometrial cancer, esophageal cancer, gallbladder cancer, genital cancer, urinary cancer Reproductive tract cancer, head cancer, kidney cancer, laryngeal cancer, liver cancer, lung cancer, muscle cancer.
  • the cancer is sarcoma, lymphoma, leukemia, melanoma, mesothelioma, multiple myeloma or seminoma.
  • diseases, disorders or conditions related to ⁇ V integrin activity include but are not limited to transplantation injections, fibrotic disorders (such as idiopathic pulmonary fibrosis, interstitial lung disease, liver Fibrosis, non-alcoholic fatty liver, primary sclerosing cholangitis (PSC), renal fibrosis, skin fibrosis, myocardial fibrosis, systemic sclerosis), inflammatory diseases (such as acute hepatitis, chronic hepatitis,, Psoriasis, irritable bowel syndrome (IBS), inflammatory bowel disease (IBD)), osteoporosis and cell proliferative diseases (such as cancer, myeloma, fibroma, liver cancer, leukemia, Kaposi's sarcoma, solid tumor).
  • fibrotic disorders such as idiopathic pulmonary fibrosis, interstitial lung disease, liver Fibrosis, non-alcoholic fatty liver, primary sclerosing cholangitis (PS
  • fibrotic diseases including but not limited to idiopathic pulmonary fibrosis (IPF), interstitial lung disease, non-specific interstitial pneumonia (NSIP) , Conventional interstitial pneumonia (UIP), radiation-induced fibrosis, familial pulmonary fibrosis, airway fibrosis, chronic obstructive pulmonary disease (COPD), diabetic nephropathy, focal segmental glomerulosclerosis , IgA nephropathy, nephropathy caused by drugs or transplantation, autoimmune nephropathy, lupus nephritis, liver fibrosis, renal fibrosis, chronic kidney disease (CKD), diabetic nephropathy (DKD), skin fibrosis, scars, systemic sclerosis , Scleroderma, viral fibrosis, non-alcoholic fatty liver disease (NAFLD), alcoholic or non-alcoholic stea
  • the present invention also provides a method for treating fibrotic diseases, inflammatory diseases or cell proliferative diseases, comprising administering a therapeutically effective amount of at least one compound to a patient in need of such treatment.
  • the "antioxidant" of the present invention alone or optionally in combination with another compound of the present invention and/or at least one other type of therapeutic agent.
  • the invention provides compounds of the invention for use in therapy.
  • the compounds of the present invention may be used in combination with additional therapeutic agents, such as one or more anti-fibrotic and/or anti-inflammatory therapeutic agents.
  • the present invention further provides a method for treating fibrotic diseases, inflammatory diseases or cell proliferative diseases, the method comprising administering to a patient in need a therapeutically effective amount of first and second therapeutic agents, wherein the first The therapeutic agent is the compound of the present invention.
  • the present invention provides a combined preparation of the compound of the present invention and another therapeutic agent for simultaneous, separate or sequential use in treatment.
  • halogen refers to F, Cl, Br, and I. In other words, F, Cl, Br, and I can be described as “halogen" in this specification.
  • aliphatic hydrocarbon group includes saturated or unsaturated, linear or branched chain or cyclic hydrocarbon groups.
  • the type of the aliphatic hydrocarbon group can be selected from alkyl, alkenyl, alkynyl, etc.
  • the carbon atoms of the aliphatic hydrocarbon group The number is preferably 1-12, and can also be 1-10, and a further preferred range is 1-6, specifically including but not limited to the following groups: methyl, ethyl, n-propyl, isopropyl, n-butyl , Isobutyl, tert-butyl, n-pentyl, isopentyl, neopentyl, n-hexyl, vinyl, 1-propenyl, 2-propenyl, 1-methylvinyl, 1-butenyl, 1-ethylvinyl, 1-methyl-2-propenyl, 2-butenyl, 3-butenyl, 2-methyl-1-propenyl, 2-
  • C 3-12 cycloalkyl should be understood to mean a saturated or unsaturated monovalent monocyclic or bicyclic ring, which has 3-12 carbon atoms, preferably C 3-8 cycloalkyl, more preferably C 3 -6 cycloalkyl.
  • a C 3-8 cycloalkyl group should be understood to mean a saturated or unsaturated monovalent monocyclic or bicyclic ring, which has 3, 4, 5, 6, 7 or 8 carbon atoms.
  • the C 3-12 cycloalkyl group may be a monocyclic hydrocarbon group, such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, cyclononyl or cyclodecyl, or a bicyclic group. Hydrocarbon groups such as tetralin or decalin.
  • 3-12 membered heterocyclic group means a saturated or unsaturated monovalent monocyclic or bicyclic ring, which contains 1-5 heteroatoms independently selected from N, O and S, and heteroatom-containing groups do not have Aromatic, the 3-12 membered heterocyclic group is preferably a 3-10 membered heterocyclic group.
  • 3-12 membered heterocyclic group means a saturated monovalent monocyclic or bicyclic ring, which contains 1-5, preferably 1-3 heteroatoms selected from N, O and S.
  • the heterocyclic group may be connected to the rest of the molecule through any one of the carbon atoms or a nitrogen atom (if present).
  • the heterocyclic group may include but is not limited to: 4-membered ring, such as azetidinyl, oxetanyl; 5-membered ring, such as tetrahydrofuranyl, tetrahydrothienyl, dioxane Pentenyl, pyrrolidinyl, imidazolidinyl, pyrazolidinyl, pyrrolinyl; or 6-membered ring, such as tetrahydropyranyl, piperidinyl, morpholinyl, dithiaalkyl, thiomorpholine Group, piperazinyl, or trithiaalkyl; or 7-membered ring, such as diazeppanyl.
  • 4-membered ring such as azetidinyl, oxetanyl
  • 5-membered ring such as tetrahydrofuranyl, tetrahydrothienyl, dioxane Penteny
  • the heterocyclic group may be benzo-fused.
  • the heterocyclic group may be bicyclic, such as but not limited to a 5, 5-membered ring, such as hexahydrocyclopenta[c]pyrrole-2(1H)-yl ring, or a 5, 6-membered bicyclic ring, such as hexahydropyrrole And [1,2-a]pyrazine-2(1H)-yl ring.
  • the ring containing the nitrogen atom may be partially unsaturated, that is, it may contain one or more double bonds, such as but not limited to 2,5-dihydro-1H-pyrrolyl, 4H-[1,3,4]thiadi Azinyl, 4,5-dihydrooxazolyl or 4H-[1,4]thiazinyl, or it may be benzo-fused, such as but not limited to dihydroisoquinolinyl, 2,3- Dihydrobenzofuranyl, 3,4-dihydro-2H-1-benzopyranyl (chromanyl), 2,3-dihydrobenzo[b][1,4]dioxanyl.
  • the 3-12 membered heterocyclic group can also be selected from, for example, the following groups:
  • C 6-20 aryl should be understood to preferably mean a monovalent aromatic or partially aromatic monocyclic, bicyclic or tricyclic hydrocarbon ring having 6-20 carbon atoms, preferably “C 6-10 aryl” .
  • the term C 6-20 aryl should be understood to preferably mean monovalent aromaticity with 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20 carbon atoms Or a partially aromatic monocyclic, bicyclic or tricyclic hydrocarbon ring, especially a ring with 6 carbon atoms (“C 6 aryl”), such as phenyl; or biphenyl, or one with 9 carbon atoms Ring (“C 9 aryl”), such as indanyl or indenyl, or a ring with 10 carbon atoms (“C 10 aryl”), such as tetrahydronaphthyl, dihydronaphthyl or naphthyl, Either a ring having 13 carbon atoms (“C 13 aryl”), such as fluor
  • 5-14 membered heteroaryl should be understood to include monovalent monocyclic, bicyclic or tricyclic aromatic ring systems having 5, 6, 7, 8, 9, 10, 11, 12, 13 or 14 ring atoms, especially 5 or 6 or 9 or 10 carbon atoms, and it contains 1 to 5, preferably 1 to 3 heteroatoms independently selected from N, O and S, and, in addition, in each In the case it can be benzo-fused.
  • the heteroaryl group is selected from thienyl, furyl, pyrrolyl, oxazolyl, thiazolyl, imidazolyl, pyrazolyl, isoxazolyl, isothiazolyl, oxadiazolyl, triazolyl, thiol Diazolyl, thio-4H-pyrazolyl, etc.
  • heterocyclic groups or heteroaryl groups include all possible isomeric forms thereof, such as positional isomers thereof. Therefore, for some illustrative non-limiting examples, pyridinyl or pyridinylene includes pyridin-2-yl, pyridin-2-yl, pyridin-3-yl, pyridin-3-yl, pyridin-4-yl And pyridin-4-yl; thienyl or thienylene includes thiophen-2-yl, thiophen-2-yl, thiophen-3-yl and thiophen-3-yl.
  • the compound of the present invention may be chiral, and therefore may exist in various enantiomeric forms. Therefore, these compounds may exist in racemate form or optically active form.
  • the compounds of the present invention or intermediates thereof can be separated into enantiomeric compounds by chemical or physical methods known to those skilled in the art, or used in synthesis in this form. In the case of racemic amines, diastereomers are prepared from the mixture by reaction with optically active resolving reagents.
  • Suitable resolution reagents are optically active acids such as R and S forms of tartaric acid, diacetyl tartaric acid, dibenzoyl tartaric acid, mandelic acid, malic acid, lactic acid, appropriate N-protected amino acids (e.g., N- Benzoyl proline or N-benzenesulfonyl proline) or various optically active camphor sulfonic acids.
  • optically active resolving reagents such as dinitrobenzoylphenylglycine, cellulose triacetate or other carbohydrate derivatives or chiral derivatized methacrylate polymers
  • Suitable eluents for this purpose are aqueous or alcohol-containing solvent mixtures, for example, hexane/isopropanol/acetonitrile.
  • N-oxides since nitrogen needs to have available lone pairs of electrons for being oxidized to nitrogen oxides, not all nitrogen-containing heterocycles can form N-oxides; those skilled in the art will recognize that N-oxides can be formed. -Nitrogen-containing heterocycles of oxides. Those skilled in the art will also recognize that tertiary amines can form N-oxides.
  • the synthetic methods for preparing heterocyclic and tertiary amine N-oxides are well known to those skilled in the art, and the synthetic methods include the use of peroxyacids such as peroxyacetic acid and m-chloroperoxybenzoic acid (MCPBA), peroxy Hydrogen oxide, alkyl hydroperoxides such as tert-butyl hydroperoxide, sodium perborate, and dioxirane such as dimethyldioxirane oxidize heterocycles and tertiary amines.
  • MCPBA m-chloroperoxybenzoic acid
  • alkyl hydroperoxides such as tert-butyl hydroperoxide
  • sodium perborate sodium perborate
  • dioxirane such as dimethyldioxirane oxidize heterocycles and tertiary amines.
  • the pharmaceutically acceptable salt may be, for example, an acid addition salt of the compound of the present invention that has a nitrogen atom in the chain or ring and is sufficiently basic, for example, an acid addition salt formed with the following inorganic acids: for example, hydrochloric acid, hydrofluorine Acid, hydrobromic acid, hydroiodic acid, sulfuric acid, pyrosulfuric acid, phosphoric acid or nitric acid, or hydrogen sulfate, or acid addition salts formed with the following organic acids: for example, formic acid, acetic acid, acetoacetic acid, pyruvic acid, trifluoroacetic acid , Propionic acid, butyric acid, caproic acid, heptanoic acid, undecanoic acid, lauric acid, benzoic acid, salicylic acid, 2-(4-hydroxybenzoyl)benzoic acid, camphor acid, cinnamic acid, cyclopentane Propionic acid, digluconic acid, 3-hydroxy-2-nap
  • an alkali metal salt such as a sodium salt or potassium salt
  • an alkaline earth metal salt such as a calcium salt or a magnesium salt
  • an ammonium salt or salts formed with organic bases that provide physiologically acceptable cations, such as salts formed with sodium ions, potassium ions, N-methylglucamine, dimethylglucamine, ethylglucosamine, Lysine, dicyclohexylamine, 1,6-hexanediamine, ethanolamine, glucosamine, meglumine, sarcosine, serinol, trihydroxymethylaminomethane, aminopropanediol, 1-amino-2 ,3,4-Butanetriol.
  • an alkali metal salt such as a sodium salt or potassium salt
  • an alkaline earth metal salt such as a calcium salt or a magnesium salt
  • an ammonium salt or salts formed with organic bases that provide physiologically acceptable cations, such as salts formed with sodium ions, potassium ions, N-methyl
  • the pharmaceutically acceptable salt includes the salt formed by the group -COOH with the following substances: sodium ion, potassium ion, calcium ion, magnesium ion, N-methylglucamine, dimethylglucamine, Ethyl glucosamine, lysine, dicyclohexylamine, 1,6-hexanediamine, ethanolamine, glucosamine, meglumine, sarcosine, serinol, trishydroxymethylaminomethane, aminopropanediol , 1-Amino-2,3,4-butanetriol.
  • basic nitrogen-containing groups can be quaternized with the following reagents: lower alkyl halides, such as methyl, ethyl, propyl, and butyl chlorides, bromides, and iodides; dialkyl sulfates, such as sulfuric acid Dimethyl, diethyl sulfate, dibutyl sulfate and dipentyl sulfate; long chain halides such as decyl, lauryl, myristyl and stearyl chlorides, bromides and iodides; aralkyl Halides such as benzyl and phenethyl bromide.
  • lower alkyl halides such as methyl, ethyl, propyl, and butyl chlorides, bromides, and iodides
  • dialkyl sulfates such as sulfuric acid Dimethyl, diethyl sulfate, dibutyl sulfate and dipentyl
  • pharmaceutically acceptable salts include hydrochloride, sulfate, nitrate, bisulfate, hydrobromide, acetate, oxalate, citrate, methanesulfonate, formate, or Meglumine salt and so on.
  • the pharmaceutically acceptable salt includes not only the salt formed on one of the salt-forming sites of the compound of the present invention, but also two, three, or all of them.
  • the salt formed on the site for this reason, the molar ratio of the compound of formula (I) to the acid radical ion (anion) or base cation required for salt formation in the pharmaceutically acceptable salt can be varied within a relatively large range, for example, it can be 4:1. ⁇ 1:4, such as 3:1, 2:1, 1:1, 1:2, 1:3, etc.
  • the compounds of the present invention may also contain one or more asymmetric centers.
  • Asymmetric carbon atoms can exist in (R) or (S) configuration. When there is only one asymmetric center, a racemic mixture is produced, and when multiple asymmetric centers are contained, a diastereomeric mixture is obtained. In some cases, there may be asymmetry due to hindered rotation around a specific bond, for example, the central bond connects two substituted aromatic rings of a specific compound.
  • the substituents may also exist in cis or trans isomeric forms.
  • the compounds of the present invention also include all possible stereoisomers of each, which is a single stereoisomer or the stereoisomer (for example, R-isomer or S-isomer, or E-isomer or Z-isomer) in the form of any mixture in any ratio.
  • a single stereoisomer (e.g., single enantiomer or single diastereomer) of the compound of the present invention can be achieved by any suitable prior art method (e.g., chromatography, especially, e.g., chiral chromatography) Separation.
  • tautomer refers to an isomer of a functional group resulting from the rapid movement of an atom in a molecule at two positions.
  • the compounds of the present invention may exhibit tautomerism.
  • Tautomeric compounds can exist in two or more mutually convertible species.
  • Proton shift tautomers result from the migration of covalently bonded hydrogen atoms between two atoms.
  • Tautomers generally exist in an equilibrium form, and an attempt to separate a single tautomer usually produces a mixture whose physical and chemical properties are consistent with a mixture of compounds. The position of equilibrium depends on the chemical properties of the molecule.
  • the ketone type is dominant; in phenol, the enol type is dominant.
  • the present invention encompasses all tautomeric forms of the compound.
  • the involved compounds also include isotopically-labeled compounds.
  • the isotopically-labeled compounds are the same as those shown in Formula I, but one or more of the atoms are different from the usual atomic mass or mass number. Naturally occurring atomic mass or mass number atomic substitution.
  • isotopes that can be incorporated into the compounds of the present invention include isotopes of H, C, N, O, S, F, and Cl, such as 2 H, 3 H, 13 C, 11 C, 14 C, 15 N, 18 O, respectively. , 17 O, 32 P, 35 S, 18 F and 36 Cl.
  • the compounds of the present invention containing the above-mentioned isotopes and/or other isotopes of other atoms, prodrugs thereof, or pharmaceutically acceptable salts of the compounds or prodrugs are within the scope of the present invention.
  • the replacement of heavier isotopes can provide certain therapeutic advantages derived from higher metabolic stability (for example, increased in vivo half-life or reduced dosage requirements), and therefore can be used in certain Some cases are preferred.
  • the compounds of the present invention as claimed in the claims can be specifically defined to be substituted with deuterium or tritium.
  • deuterium or tritium is not separately listed for the hydrogen in the substituent does not mean that deuterium or tritium is excluded, but deuterium or tritium may also be included in the same way.
  • the term "effective amount” or “therapeutically effective amount” refers to the amount of the compound of the present invention sufficient to achieve the intended application (including but not limited to the treatment of diseases as defined below).
  • the therapeutically effective amount may vary depending on the following factors: the intended application (in vitro or in vivo), or the subject to be treated and the disease condition such as the weight and age of the subject, the severity of the disease condition and the mode of administration, etc. It can be easily determined by a person of ordinary skill in the art.
  • the specific dosage will vary depending on the following factors: the particular compound selected, the dosing regimen on which it is based, whether it is administered in combination with other compounds, the timing of administration, the tissue to be administered, and the physical delivery system carried.
  • solvate refers to those forms of the compound of the present invention, which form a complex by coordination with solvent molecules in a solid or liquid state. Hydrates are a specific form of solvates in which the coordination is carried out with water. In the present invention, the preferred solvate is a hydrate. Further, the pharmaceutically acceptable solvate (hydrate) of the compound of the general formula I of the present invention refers to the co-crystal and clathrate formed by the compound I and one or more stoichiometric molecules of water or other solvents. Solvents that can be used for solvates include, but are not limited to: water, methanol, ethanol, ethylene glycol, and acetic acid.
  • prodrug represents the conversion of a compound into a compound represented by the aforementioned general formula or specific compound in vivo. Such conversion is affected by the hydrolysis of the prodrug in the blood or the enzymatic conversion of the prodrug into the maternal structure in the blood or tissues.
  • the prodrugs of the present invention can be esters.
  • esters can be used as prodrugs including phenyl esters, aliphatic esters, acyloxymethyl esters, carbonates, carbamates and amino acid esters.
  • a compound in the present invention contains a hydroxyl/carboxyl group, that is, it can be acylated to obtain a compound in the form of a prodrug.
  • Other prodrug forms include phosphate esters, such as these phosphate ester compounds are obtained by phosphorylation of the parent hydroxyl group.
  • the present invention provides a novel compound of general formula I, which has a good inhibitory effect on integrins;
  • the compounds of the present invention have inhibitory effects on multiple integrin subtypes, some of which have obvious inhibitory effects on one or more of ⁇ V ⁇ 1, ⁇ V ⁇ 3, ⁇ V ⁇ 5, ⁇ V ⁇ 6, and ⁇ V ⁇ 8, ⁇ 5 ⁇ 1, and ⁇ 8 ⁇ 1.
  • Deuterated reagents (DMSO-d 6 , CDCl 3 , CD 3 OD) were used to record 1 H and 19 F NMR on a Bruker Ascend 400 spectrometer. Deuterated solvent or TMS is used as internal standard. The chemical shift is expressed in ppm, the coupling constant (J) is expressed in Hz, and the splitting methods are s (single peak), d (double peak), t (triplet), q (quartet), m (multiple peak), br (broad peak).
  • LC-MS uses Agilent 1260-6120 system with Waters Cortecs C18, 2.7 ⁇ m, 4.6 ⁇ 30mm column; HPLC uses Waters Acquity UPLC H-class instrument with Acquity BEH C18, 1.7 ⁇ m, 50 ⁇ 2.1mm column.
  • Step 1 7-Methyl-1,2,3,4-tetrahydro-1,8-naphthyridine
  • Step 2 7-Methyl-3,4-dihydro-1,8-naphthyridine-1(2H)-carboxylic acid tert-butyl ester
  • Step 3 7-(2-Methoxy-2-oxoethyl)-3,4-dihydro-1,8-naphthyridine-1(2H)-carboxylic acid tert-butyl ester
  • Step 4 7-(2-hydroxyethyl)-3,4-dihydro-1,8-naphthyridine-1(2H)-carboxylic acid tert-butyl ester
  • Step 5 7-(2-(Tosyloxy)ethyl)-3,4-dihydro-1,8-naphthyridine-1(2H)-carboxylic acid tert-butyl ester
  • Step 4 7-(5-Methoxy-5-oxopentyl)-3,4-dihydro-1,8-naphthyridine-1(2H)-carboxylic acid tert-butyl ester
  • Step 5 5-(8-(tert-Butoxycarbonyl)-5,6,7,8-tetrahydro-1,8-naphthyridin-2-yl)pentanoic acid
  • Step 1 7-(5-(Methoxy(methyl)amino)-5-oxopentyl)-3,4-dihydro-1,8-naphthyridine-1(2H)-carboxylic acid tert-butyl ester
  • Step 2 7-(5-oxapentyl)-3,4-dihydro-1,8-naphthyridine-1(2H)-carboxylic acid tert-butyl ester
  • Step 1 (S)-3-(((Benzyloxy)carbonyl)amino)pyrrolidine-1-carboxylic acid tert-butyl ester
  • Step 2 Synthesis of (S)-3-(((benzyloxy)carbonyl)(methyl)amino)pyrrolidine-1-carboxylic acid tert-butyl ester
  • Step 4 (S)-7-(5-(3-((((benzyloxy)carbonyl)(methyl)amino)pyrrolidin-1-yl)pentyl)-3,4-dihydro-1, 8-naphthyridine-1(2H)-tert-butyl carboxylate
  • Step 5 (S)-7-(5-(3-(methylamino)pyrrolidin-1-yl)pentyl)-3,4-dihydro-1,8-naphthyridine-1(2H)- Tert-butyl carboxylate
  • the intermediate Int-F is synthesized:
  • the intermediate Int-G is synthesized:
  • the intermediate Int-H is synthesized:
  • Step 1 tert-Butyl 3-(((R)-2-methoxy-2-oxo-1-phenethyl)amino)pyrrolidin-1-carboxylate
  • Step 2 3-(((R)-2-methoxy-2-oxo-1-phenylethyl)(methyl)amino)pyrrolidine-1-carboxylic acid tert-butyl ester
  • Step 1 7-(3-(3-(((R)-2-methoxy-2-oxo-1-phenylethyl)amino)pyrrolidin-1-yl)propyl)-3,4 -Dihydro-1,8-naphthyridine-1(2H)-carboxylic acid tert-butyl ester
  • Step 1 (3S)-3-((2-Methoxy-2-oxo-1-phenylethyl)amino)pyrrolidine-1-carboxylic acid tert-butyl ester
  • Step 2 (3S)-3-((2-Methoxy-2-oxo-1-phenylethyl)(methyl)amino)pyrrolidine-1-carboxylate tert-butyl ester
  • Step 4 7-(3-((3S)-3-((2-methoxy-2-oxo-1-phenethyl)(methyl)amino)pyrrolidin-1-yl)propyl)- 3,4-Dihydro-1,8-naphthyridine-1(2H)-carboxylic acid tert-butyl ester
  • Step 5 2-(Methyl((S)-1-(3-(5,6,7,8-tetrahydro-1,8-naphthyridin-2-yl)propyl)pyrrolidin-3-yl )Amino)-2-phenylacetic acid
  • Example 4 The following compounds can be obtained by referring to the synthetic methods of compound 1 and compound 5:
  • the compound 10 was prepared, and the diastereomeric mixture was separated by preparing chiral SFC under the following conditions: [Column: Chiralpak AD-3 (50 ⁇ 4.6mm), mobile phase: 40% ethanol in CO 2 solution (containing 0.05% DEA), flow rate: 3 mL/min] to obtain compound 10a and compound 10b as white solids, respectively.
  • the compound 11 was prepared, and the mixture of diastereomers was separated by preparing chiral SFC under the following conditions: [Column: (S, S) Whelk-O1 (100 ⁇ 4.6mm); mobile phase: 40% methanol in carbon dioxide Solution (containing 0.05% DEA); flow rate: 3 mL/min] to obtain compound 11a and compound 11b, respectively, as white solids.
  • the compound 12 was prepared. By preparing chiral SFC, the mixture of diastereomers was separated under the following conditions [column: Chiralpak IC-3 (50 ⁇ 4.6mm); mobile phase: 40% methanol in carbon dioxide (containing 0.05%) DEA); flow rate: 3 mL/min], compound 12a and compound 12b were obtained as white solids, respectively.
  • the compound 13 was prepared, and the crude residue was separated by preparing chiral SFC under the following conditions [column: Diacel ChiralPak IG (250 ⁇ 30 mm, 10 ⁇ m). Mobile phase: 60% methanol in carbon dioxide solution (containing 0.1% NH 3 .H 2 O); flow rate: 70 g/min] to obtain compound 13a and compound 13b, respectively, as light yellow solids.
  • the compound 14 was prepared, and the crude residue was separated by preparing chiral SFC under the following conditions: [Column: Daicel ChiralPak IG (250 ⁇ 30 mm, 10 ⁇ m); column chromatography. Mobile phase: 60% methanol in carbon dioxide solution (containing 0.1% NH 3 .H 2 O) flow rate: 70 g/min] to obtain compound 14a and compound 14b, respectively, as light yellow solids.
  • Step 1 7-(3-(3-(((R)-2-methoxy-2-oxo-1-phenylethyl)(methyl)amino)pyrrolidin-1-yl)-3- (Oxypropyl)-3,4-dihydro-1,8-naphthyridine-1(2H)-carboxylic acid tert-butyl ester
  • Step 2 (2R)-2-(Methyl(1-(3-(5,6,7,8-tetrahydro-1,8-naphthyridin-2-yl)propionyl)pyrrolidin-3-yl )Amino)-2-phenylmethyl acetate
  • Step 3 (2R)-2-(Methyl(1-(3-(5,6,7,8-tetrahydro-1,8-naphthyridin-2-yl)propionyl)pyrrolidin-3-yl )Amino)-2-phenylacetic acid
  • Step 1 tert-Butyl 3-(phenylamino)pyrrolidine-1-carboxylate
  • Step 2 tert-Butyl 3-((2-methoxy-2-oxoethyl)(phenyl)amino)pyrrolidine-1-carboxylate
  • Step 4 7-(3-(3-((2-methoxy-2-oxoethyl)(phenyl)amino)pyrrolidin-1-yl)propyl)-3,4-dihydro-1 ,8-naphthyridine-1(2H)-carboxylic acid tert-butyl ester
  • Step 5 N-Phenyl-N-(1-(3-(5,6,7,8-tetrahydro-1,8-naphthyridin-2-yl)propyl)pyrrolidin-3-yl) Glycine Methyl Ester
  • Step 6 N-Phenyl-N-(1-(3-(5,6,7,8-tetrahydro-1,8-naphthyridin-2-yl)propyl)pyrrolidin-3-yl)glycine
  • Step 1 tert-Butyl 3-(benzyl(2-ethoxy-2-oxyethyl)amino)pyrrolidine-1-carboxylate
  • Step 1 (3R)-3-((1-(3-chlorophenyl)-3-methoxy-3-oxopropyl)amino)pyrrolidine-1-carboxylic acid tert-butyl ester
  • Step 3 7-(2-((3R)-3-((1-(3-chlorophenyl)-3-methoxy-3-oxopropyl)amino)pyrrolidin-1-yl)ethyl )-3,4-dihydro-1,8-naphthyridine-1(2H)-carboxylic acid tert-butyl ester
  • Step 4 3-(3-Chlorophenyl)-3-(((R)-1-(2-(5,6,7,8-tetrahydro-1,8-naphthyridin-2-yl)ethyl (Yl)pyrrolidone-3-yl)amino)methyl propionate
  • Step 5 3-(3-Chlorophenyl)-3-(((R)-1-(2-(5,6,7,8-tetrahydro-1,8-naphthyridin-2-yl)ethyl (Yl)pyrrolidone 3-yl)amino)propionic acid
  • the crude product was purified by preparative HPLC (column: Xbridge 5u C18 150 ⁇ 19mm; mobile phase: acetonitrile-water (0.05% formic acid); gradient: 5-15% acetonitrile; flow rate: 20 mL/min) to obtain the title compound (0.12g , 32%), is a yellow solid.
  • Compound 27 was prepared, and the crude product was purified by preparative HPLC (column: Kromasil-C18 100 ⁇ 21.2mm 5um; mobile phase: acetonitrile-water (0.1% formic acid); gradient: 10-20%) to obtain compound 27a and compound 27b.
  • the compound 28 was prepared, and the crude product was purified by preparative HPLC (column: Kromasil-C18 100 ⁇ 21.2mm 5um; mobile phase: acetonitrile-water (0.1% formic acid); gradient: 10-20%) to obtain the compound as a yellow solid 28a and compound 28b.
  • Step 3-7 Refer to steps 1-5 of the synthesis of compound 23, and chiral separation of 3-(3-iso) by SFC (column: chiralpak-IG; mobile phase: CO 2 -methanol (containing 0.1% DEA)) Propoxyphenyl)-3-(((R)-1-(2-(5,6,7,8-tetrahydro-1,8-naphthyridin-2-yl)ethyl)pyrrolidine-3 -Yl)amino)propionic acid (30 mg, 0.07 mmol) to give compound 29a and compound 29b.
  • the compound 33 was chiral separated by SFC (column: Chiralpak-IG; mobile phase: CO 2 -methanol (0.1% DEA)) to give compound 33a and compound 33b.
  • Compound 39 was purified by preparative HPLC (column: Xtimate 10u C18 250 ⁇ 30; mobile phase: acetonitrile-water (0.1% formic acid); gradient: 10-20%, 9 minutes; flow rate: 50 mL/min) to obtain Compound 39a and Compound 39b.
  • Compound 46 was prepared. The crude product was purified by preparative HPLC (column: Xbridge 5u C18 150 ⁇ 19mm; mobile phase: acetonitrile-water (0.05% formic acid); gradient: 5-15% acetonitrile, flow rate: 20 mL/min) to obtain Compound 46a and Compound 46b.
  • Compound 49 was prepared and purified by preparative HPLC (column: Kromasil-C18 100 ⁇ 21.2mm 5um; mobile phase: acetonitrile-water (0.1% formic acid); gradient: 10-20% acetonitrile) to obtain compound 49a and compound 49b .
  • Compound 51 was prepared and purified by preparative HPLC (column: Xbridge 5u C18 150 ⁇ 19mm; mobile phase: acetonitrile-water (0.05% formic acid); gradient: 5-15% acetonitrile, flow rate: 20 mL/min).
  • Chromatin-based Chromatography columnar phase: aqueous quaternary ammonium salt
  • Compound 51b was prepared and purified by preparative HPLC (column: Xbridge 5u C18 150 ⁇ 19mm; mobile phase: acetonitrile-water (0.05% formic acid); gradient: 5-15% acetonitrile, flow rate: 20 mL/min).
  • Compound 51a and Compound 51b were prepared and purified by preparative HPLC (column: Xbridge 5u C18 150 ⁇ 19mm; mobile phase: acetonitrile-water (0.05% formic acid); gradient: 5-15% acetonitrile, flow rate: 20 mL/min).
  • Compound 55 was prepared and purified by preparative HPLC (column: Kromasil-C18 100 ⁇ 21.2mm 5um; mobile phase: acetonitrile-water (0.1% formic acid); gradient: 2-8% acetonitrile) to obtain compound 55a and compound 55b .
  • Step 1 1-(3-Bromophenyl)-3-methyl-1H-pyrazole (a) and 1-(3-bromophenyl)-5-methyl-1H-pyrazole (b)
  • Step 4-8 follow steps 1-5 of compound 23 to synthesize to obtain compound 63 (formate):
  • Example 14 With reference to the synthetic method of compound 63, the following compounds were prepared:
  • Compound 70 was prepared, and the mixture was purified by preparative HPLC (column: Kromasil C18 5um 100 ⁇ 21.5mm; mobile phase: acetonitrile-water (0.1% formic acid); gradient: 89-99% acetonitrile, 6.5 minutes; flow rate: 25 mL /min) to obtain compound 70a and compound 70b.
  • Triethylamine (7.5 mL, 54 mmol) was added to the solution at 25°C, and then stirred for 1 hour.
  • Compound 91 was prepared and purified by preparative HPLC (column: Kromasil-C18 100 ⁇ 21.2mm 5um; mobile phase: acetonitrile-water (0.1% trifluoroacetic acid); gradient: 25-35%) to obtain compound 91a and compound 91b.
  • Step 5 (S)-3-(((R)-tert-butylsulfinyl)amino)-3-(3-(3,5-dimethyl-1H-pyrazol-1-yl)phenyl ) Ethyl Propionate
  • reaction mixture was cooled to 0°C, and N-(3-(3,5-dimethyl-1H-pyrazol-1-yl)benzylidene)-2-methylpropane-2-sulfenamide (A solution of 2.0 g 6.7 mmol) in tetrahydrofuran (10 mL) was added dropwise to the reaction solution. After stirring at 0°C for 3 hours, the reaction solution was filtered through Celite. The filtrate was washed with 1N hydrochloric acid aqueous solution, NaHCO 3 aqueous solution, brine successively, dried over sodium sulfate and filtered, and the filtrate was removed in vacuo to obtain the crude compound (1.9 g, 76%) as a yellow oil.
  • Step 7 3-(((S)-1-(3-(3,5-Dimethyl-1H-pyrazol-1-yl)phenyl)-3-ethoxy-3-oxopropyl) Amino)pyrrolidine-1-tert-butyl carboxylate
  • Step 8 (3S)-3-(3-(3,5-Dimethyl-1H-pyrazol-1-yl)phenyl)-3-(pyrrolidin-3-ylamino)propionic acid ethyl ester
  • Step 9 7-(2-(3-(((S)-1-(3-(3,5-dimethyl-1H-pyrazol-1-yl)phenyl)phenyl)-3-ethyl (Oxy-3-oxopropyl)amino)pyrrolidin-1-yl)ethyl)-3,4-dihydro-1,8-naphthyridine-1(2H)-carboxylic acid tert-butyl ester
  • Steps 10-11 (3S)-3-(3-(3,5-dimethyl-1H-pyrazol-1-yl)phenyl)-3-((1-(2-(5,6, 7,8-Tetrahydro-1,8-naphthyridin-2-yl)ethyl)pyrrolidin-3-yl)amino)propionic acid
  • Step 2 (3S)-3-((1-(3-(3,5-dimethyl-1H-pyrazol-1-yl)phenyl)-3-methoxy-3-oxopropyl) Amino) pyrrolidine-1-carboxylic acid tert-butyl ester
  • Step 3 Methyl 3-(3-(3,5-dimethyl-1H-pyrazol-1-yl)phenyl)-3-(((S)-pyrrolidin-3-yl)amino)propionate ester
  • Step 4 7-(2-((3S)-3-((1-(3-(3,5-dimethyl-1H-pyrazol-1-yl)phenyl)-3--3-methyl (Oxy)amino)pyrrolidin-1-yl)ethyl)-3,4-dihydro-1,8-naphthyridine-1(2H)-carboxylic acid tert-butyl ester
  • Step 5 3-(3-(3,5-Dimethyl-1H-pyrazol-1-yl)phenyl)-3-(((S)-1-(2-(5,6,7, 8-tetrahydro-1,8-naphthyridin-2-yl)ethylpyrrolidin-3-yl)amino)propionic acid
  • Compound 105 was prepared, and the crude product was separated by preparing chiral SFC under the following conditions: [Column: Daicel ChiralPak IG (250 ⁇ 30mm, 10 ⁇ m); Mobile phase: 60% methanol in carbon dioxide solution (0.1% NH 3 .H 2 O); flow rate: 70 g/min], compound 105a and compound 105b were obtained as white solids, respectively.
  • Formaldehyde (2.1g, 70mmol) and potassium tert-butoxide (1.5g, 13.33mmol) were added to a solution of methyl 2-phenylacetate (10.0g, 66.6mmol) in DMSO (150mL), and the reaction solution was reacted at room temperature for 16 After hours, ice water (100mL) was poured into the reaction solution, 1N hydrochloric acid was added dropwise to adjust pH ⁇ 5, the resulting mixture was diluted with water (100mL), extracted with ethyl acetate (150mL ⁇ 2), and the combined organic phase was saturated with NaHCO 3 (20 mL) and brine, and dried over sodium sulfate.
  • Step 3 (3R)-3-((3-Methoxy-3-oxo-2-phenylpropyl)amino)pyrrolidine-1-carboxylic acid tert-butyl ester
  • Steps 4-7 were carried out with reference to the synthesis method of compound 97 steps 9-11, and compound 108 (formate) was prepared: 1 HNMR (400MHz, CD 3 OD) ⁇ 7.33-7.28(m,5H),7.26-7.24( m, 1H), 6.51-6.49 (m, 1H), 3.79-3.77 (m, 1H), 3.74-3.71 (m, 1H), 3.47-3.42 (m, 2H), 3.40-3.33 (m, 2H), 3.32-3.30 (m, 2H), 3.27-3.11 (m, 2H), 3.04-3.01 (m, 2H), 2.87-2.58 (m, 4H), 2.31-2.30 (m, 1H), 2.27-2.03 (m ,1H),1.96-1.85(m,2H).
  • LC-MS ESI m/z 395.1[M+H] + , C 23 H 30 N 4 O 2. Calculated value 394.24.
  • Example 21 With reference to the synthetic method of compound 108, the following compounds were prepared:
  • Step 1 tert-Butyl 3-((3-ethoxy-3-oxoprop-1-en-1-yl)oxy)pyrrolidine-1-carboxylate
  • Step 2 3-((1-(3-(3,5-Dimethyl-1H-pyrazol-1-yl)phenyl)-3-ethoxy-3-oxoprop-1-ene- 1-yl)oxy)pyrrolidine-1-carboxylic acid tert-butyl ester
  • Step 3 3-(1-(3-(3,5-Dimethyl-1H-pyrazol-1-yl)phenyl)-3-ethoxy-3-oxopropoxy)pyrrolidine-1 -Tert-butyl formate
  • Step 4-7 Refer to the synthesis of compound 23 (Step 2-5) to prepare compound 112: 1 H NMR (400MHz, DMSO-d 6 , 80°C) ⁇ 7.48–7.42(m,2H), 7.39– 7.33(m,2H),7.03-6.97(m,1H),6.31-6.23(m,1H),6.05(s,1H),5.94(brs,1H),4.84-4.76(m,1H),3.95( s, 1H), 3.31-3.22(m, 2H), 2.77-2.53(m, 10H), 2.47-2.33(m, 2H), 2.29(s, 3H), 2.19(s, 3H), 1.86-1.74( m,3H),1.59–1.57(m,1H).
  • LC-MS ESI m/z 490.3[M+H] + ; C 28 H 35 N 5 O 3 calculated value 489.27.
  • HPLC purity 100.0% (214nm) , 100.0% (254
  • Step 2 3-((3-Methoxy-3-oxo-1-phenylprop-1-en-1-yl)amino)-3-methylpyrrolidine-1-carboxylic acid tert-butyl ester
  • Step 3 3-((3-Methoxy-3-oxo-1-phenylpropyl)amino)-3-methylpyrrolidine-1-carboxylic acid tert-butyl ester
  • Step 4 3-((3-Methylpyrrolidin-3-yl)amino)-3-phenylpropionic acid methyl ester
  • Step 5 7-(2-(3-((3-methoxy-3-oxo-1-phenylpropyl)amino)-3-methylpyrrolidin-1-yl)ethyl)-3 ,4-Dihydro-1,8-naphthyridine-1(2H)-carboxylic acid tert-butyl ester
  • Step 6 Methyl 3-((3-methyl-1-(2-(5,6,7,8-tetrahydro-1,8-naphthyridin-2-yl)ethyl)pyrrolidine-3- (Yl)amino)-3-phenylpropionate
  • Step 7 3-((3-methyl-1-(2-(5,6,7,8-tetrahydro-1,8-naphthyridin-2-yl)ethyl)pyrrolidin-3-yl) Amino)-3-phenylpropionic acid
  • the crude product was purified by preparative HPLC (column: Xbridge 5u C18 150 ⁇ 19mm; mobile phase: acetonitrile-water (0.05% NH 3 ⁇ H 2 O); gradient: 5-15% acetonitrile; flow rate: 20 mL/min) to obtain the title
  • the compound (6 mg, 4.3%) was a yellow solid.
  • Step 1 tert-Butyl 3-(((R)-2-methoxy-2-oxo-1-phenylethyl)carbamoyl)pyrrolidine-1-carboxylate
  • Step 3 7-(2-(3-(((R)-2-methoxy-2-oxo-1-phenylethyl)carbamoyl)pyrrolidin-1-yl)-2-oxy (Ethyl)-3,4-dihydro-1,8-naphthyridine-1(2H)-carboxylic acid tert-butyl ester
  • Step 4 (2R)-2-Phenyl-2-(1-(2-(5,6,7,8-tetrahydro-1,8-naphthyridin-2-yl)acetyl)pyrrolidine-3 -Carboxamido) methyl acetate hydrochloride
  • Step 5 (2R)-2-Phenyl-2-(1-(2-(5,6,7,8-tetrahydro-1,8-naphthyridin-2-yl)acetyl)pyrrolidine-3 -Carboxamido) acetic acid
  • the aqueous phase was lyophilized, and the crude product obtained was purified by preparative HPLC (column: Kromasil-C18 100 ⁇ 21.2mm 5um; mobile phase: acetonitrile-water (0.05% NH 3 .H 2 O); gradient: 10-20%; flow rate: 20 mL/min) to obtain the title compound (8.9 mg, 2%) as a white solid.
  • Example 25 With reference to the synthesis of compound 114, compound 23, and compound 16, the following compounds can be prepared respectively:
  • Step 1 tert-Butyl 3-((((R)-2-methoxy-2-oxo-1-phenylethyl)amino)methyl)pyrrolidine-1-carboxylate
  • Step 2 3-((((R)-2-methoxy-2-oxo-1-phenylethyl)(methyl)amino)methyl)pyrrolidine-1-carboxylic acid tert-butyl
  • Step 4-6 Refer to step 3-5 of compound 114 to synthesize and prepare compound 125 (ammonium salt).
  • 1 H NMR 400MHz, DMSO-d 6, 80°C) ⁇ 8.13(s, 1H), 7.37-7.28( m,5H), 7.03--7.00(m,1H), 6.29--6.27(m,1H), 5.96(brs,1H), 4.19--4.14(m,1H), 3.43--3.28(m,2H), 3.26--- 2.88 (m, 2H), 2.68-2.54 (m, 5H), 2.48-2.31 (m, 6H), 2.20-2.18 (m, 3H), 1.93-1.88 (m, 1H), 1.79-1.74 (m, 2H) ),1.62–1.46(m,1H).
  • LC-MS (ESI)m/z 437.4[M+H] + ; C 25 H 32 N 4 O 3 calculated value 436.25.
  • HPLC Purity 99.7% (254
  • Example 27 With reference to the synthesis of compound 125, the following compounds were prepared:
  • the compound 134 mixture was purified by preparative HPLC (column: Kromasil-C18 100 ⁇ 21.2mm 5um, mobile phase: acetonitrile-water (0.1% formic acid); gradient: 10-20%) to obtain peak 1 compound (18 mg, 28 %) and peak 2 compound (19.5 mg, 30%).
  • Step 2 tert-Butyl 3-((((R)-2-methoxy-2-oxo-1-phenethyl)(methyl)carbamoyl)pyrrolidine-1-carboxylate
  • Step 4-6 Refer to the step 3-5 of compound 114 to synthesize and prepare compound 140 (formate).
  • LC-MS (ESI) m/z 451.3[M+H] + ; C 25 H 30 N 4 O 4 calculated value 450.23.
  • HPLC purity 98.6% (214nm), 98.6% (254nm) .
  • Example 29 With reference to the synthesis of compound 140, the following compounds were prepared:
  • Step 3 (3S)-3-((2-Methoxy-2-oxo-1-(2-(trifluoromethyl)phenyl)ethyl)amino)pyrrolidine-1-carboxylic acid tert-butyl ester
  • Step 4-7 Referring to the synthesis method of step 2-5 of compound 5, compound 146 (formate) (29.8 mg, 13%) was prepared as a white solid.
  • 1 H NMR 400MHz, CD 3 OD
  • 7.34(d,J 7.3Hz,1H)
  • 6.44(d,J 7.3Hz,1H)
  • Example 31 With reference to the synthesis method of compound 146, the following compounds were prepared:
  • Step 2 7-(5-((3S)-3-((1-(3-chlorophenyl)-2-methoxy-2-oxoethyl)(methyl)amino)pyrrolidine-1- (Yl)pentyl)-3,4-dihydro-1,8-naphthyridine-1(2H)-carboxylic acid tert-butyl ester
  • Step 3 2-(3-Chlorophenyl)-2-(methyl((S)-1-(5-(5,6,7,8-tetrahydro-1,8-naphthyridin-2-yl )Amyl)pyrrolidone-3-yl)amino)acetic acid
  • Example 33 With reference to the synthesis method of compound 149, the following compounds were prepared:
  • Compound 158 was purified by preparative HPLC under the following conditions [column: Kromasil 100-5-C18, 30 ⁇ 150mm; mobile phase: 1-100% acetonitrile in water (containing 0.1% formic acid) within 14 minutes] to obtain compound 158a And compound 158b, a white solid.
  • the diastereoisomer mixture compound 159 (30mg, 0.063mmol) was prepared by SFC chiral separation (Chirex S-VAL, R-NEA, 250 ⁇ 4.6mm), using 40% methanol in carbon dioxide solution (0.05% DEA) Eluted and concentrated at a rate of 3mL/min to give compound 159a and compound 159b.
  • compound 166 (20mg, 0.04mmol) was separated by preparative chiral SFC [column: Daicel ChiralPak IG (250 ⁇ 30mm, 10um); mobile phase: 0.1% NH 3 .H 2 O in methanol; gradient : 60% methanol, 6.5 min; flow rate: 70 g/min] to obtain compound 166a and compound 166b as yellow solids, respectively.
  • the epimer mixture compound 177 (15.8mg, 0.032mmol) was separated by preparative chiral SFC [Column: Daicel ChiralPak IG (250 ⁇ 30mm, 10um); Mobile phase: 60% methanol Carbon dioxide solution (containing 0.1% NH 3 .H 2 O); flow rate: 70 g/min] to obtain compound 177a and compound 177b, respectively, as yellow solids.
  • Step 5 7-(5-((3S)-3-((2-methoxy-1-(6-methoxypyridin-3-yl)-2-oxoethyl)(methyl)amino )Pyrrolidin-1-yl)pentyl)-3,4-dihydro-1,8-naphthyridine-1(2H)-carboxylic acid tert-butyl ester
  • Step 6 Compound 224 and Compound 225
  • Example 35 With reference to the synthesis method of compound 224, the following compounds were prepared:
  • Step 3 tert-Butyl 3-((3-(3,5-dimethyl-1H-pyrazol-1-yl)phenyl)carbamoyl)pyrrolidine-1-carboxylate
  • Step 4 3-((3-(3,5-Dimethyl-1H-pyrazol-1-yl)phenyl)(2-methoxy-2-oxoethyl)carbamoyl)pyrrolidine- Tert-Butyl 1-formate
  • Steps 5-8 refer to the synthesis method of compound 97 (steps 8-11) to prepare compound 230:
  • Example 37 Resolution of compound 230 by preparative chiral SFC under the following conditions: [Column: Daicel ChiralPak IG (250*30mm, 10um); Mobile phase: 0.1% NH 3 .H 2 O in methanol; Gradient: 60% methanol, 6.5min; flow rate: 70g/min] to obtain compound 230a and compound 230b respectively, and refer to the synthesis method of compound 230 to prepare compound 231, compound 231a, compound 231b, compound 232, compound 232a, compound 232b
  • the racemic mixture compound 231 (41.14mg, 0.080mmol) was separated by chiral preparation SFC under the following conditions [column: Daicel ChiralPak IG (250*30mm, 10um); mobile phase: containing 0.1% NH 3 ⁇ H 2 O Methanol solution; gradient: 60% methanol, 6.5 min; flow rate: 70 g/min] to obtain compound 231a and compound 231b, respectively, as white solids.
  • the racemic mixture of compound 232 was separated by chiral preparation SFC under the following conditions [column: Daicel ChiralPak IG (250 ⁇ 30mm, 10um); mobile phase: containing 0.1% NH 3 ⁇ H 2 O Methanol solution; gradient: 60% methanol, 6.5 min; flow rate: 70 g/min] to obtain compound 232a and compound 232b, respectively, as white solids.
  • Step 1 (3-(3,5-Dimethyl-1H-pyrazol-1-yl)phenyl)glycine methyl ester
  • Step 2 3-(((3-(3,5-Dimethyl-1H-pyrazol-1-yl)phenyl)(2-methoxy-2-oxoethyl)amino)methyl)pyrrole Tert-butyl alkane-1-carboxylate
  • Steps 3-6 are carried out with reference to the synthesis method of compound 97 (steps 8-11) to prepare compound 238 (formate):
  • Example 39 With reference to the synthesis method of compound 238, compound 239 (formate) was prepared:
  • Step 1 3-((3-(3,5-Dimethyl-1H-pyrazol-1-yl)phenyl)(3-ethoxy-3-oxopropyl)carbamoyl)pyrrolidine- Tert-Butyl 1-formate
  • Step 2 3-(((3-(3,5-Dimethyl-1H-pyrazol-1-yl)phenyl)(2-methoxy-2-oxoethyl)amino)methyl)pyrrole Tert-butyl alkane-1-carboxylate
  • Steps 3-5 refer to the synthetic method of compound 97 (steps 9-11) to prepare compound 240 (formates):
  • Step 1 (S)-3-((2-Methoxyphenyl)carbamoyl)pyrrolidine-1-carboxylic acid tert-butyl ester
  • Step 2 (S)-3-((2-Methoxy-2-oxoethyl)(2-methoxyphenyl)carbamoyl)pyrrolidine-1-carboxylic acid tert-butyl ester
  • Step 4 (S)-7-(3-(3-((2-methoxy-2-oxoethyl)(2-methoxyphenyl)carbamoyl)pyrrolidin-1-yl) Propyl)-3,4-dihydro-1,8-naphthyridine-1(2H)-carboxylic acid tert-butyl ester

Landscapes

  • Health & Medical Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Organic Chemistry (AREA)
  • Medicinal Chemistry (AREA)
  • Veterinary Medicine (AREA)
  • Public Health (AREA)
  • General Health & Medical Sciences (AREA)
  • Animal Behavior & Ethology (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Pharmacology & Pharmacy (AREA)
  • General Chemical & Material Sciences (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Engineering & Computer Science (AREA)
  • Cardiology (AREA)
  • Physical Education & Sports Medicine (AREA)
  • Diabetes (AREA)
  • Urology & Nephrology (AREA)
  • Rheumatology (AREA)
  • Ophthalmology & Optometry (AREA)
  • Heart & Thoracic Surgery (AREA)
  • Epidemiology (AREA)
  • Pain & Pain Management (AREA)
  • Neurology (AREA)
  • Orthopedic Medicine & Surgery (AREA)
  • Hematology (AREA)
  • Biomedical Technology (AREA)
  • Dermatology (AREA)
  • Neurosurgery (AREA)
  • Gastroenterology & Hepatology (AREA)
  • Hospice & Palliative Care (AREA)
  • Oncology (AREA)
  • Vascular Medicine (AREA)
  • Pulmonology (AREA)
  • Emergency Medicine (AREA)
  • Endocrinology (AREA)
  • Obesity (AREA)
  • Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)

Abstract

一种式I所示的化合物及其消旋体、立体异构体、互变异构体、同位素标记物、氮氧化物、溶剂化物、多晶型物、代谢产物、酯、前药或其药学上可接受的盐，以及包含其的药物组合物，其制备方法，及其医药用途，所述式I结构如下：

Description

一种吡咯烷类整合素调节剂及其用途
本申请要求2020年3月6日向中国国家知识产权局提交的，专利申请号为202010153014.2，发明名称为“一种吡咯烷类整合素调节剂及其用途”在先申请的优先权。该申请的全文通过引用的方式结合于本申请中。
技术领域
本发明属于药物化合物领域，具体涉及一种吡咯烷类整合素调节剂及其用途。
背景技术
整合素家族是一类分布广泛的跨膜糖蛋白，是连接细胞外基质环境与细胞内信号传导的纽带。整合素由α亚单位和β亚单位通过非共价键连接成异二聚体，目前已发现18种α亚基和8种β亚基，可组合成至少24种整合素二聚体。作为跨膜蛋白，细胞黏附分子整合素家族是细胞外基质、炎症细胞、成纤维细胞和实质细胞之间的主要连接物质，与组织纤维化的发生、维持和发展密切相关，在组织细胞纤维化过程中介导许多关键的细胞-细胞和细胞-细胞外基质相互作用。(Manninen et al,Proteomics,2017,17(3-4):1600022.)。
在整合素家族中，αV家族主要参与机体组织的纤维化进程。整合素αV家族包括5种亚型(αVβ1、αVβ3、αVβ5、αVβ6、αVβ8)，在多种组织的正常细胞中低表达，而在纤维性组织细胞中高表达。TGF-β是参与组织纤维化形成的重要因子，尤其是TGF-β1。在炎症和纤维化中，TGF-β1参与生理修复和胶原蛋白的积累。它可以促进上皮细胞和间质细胞的胶原和纤维连接素的生成，导致在创伤修复和纤维化过程中细胞外基质(ECM)积累(Weiskirchen et al,Molecular aspects of medicine,2019,65:2.)。整合素αV家族蛋白参与激活潜藏的TGF-β分子，通过活化TGF-β诱导过度的自身免疫反应和炎症反应，促进组织纤维化进程。
不同亚型的整合素参与机体不同部位的纤维化进展，在肾组织和肺组织纤维化中αvβ6及αvβ1发挥主要作用，在心脏纤维化中αvβ3/5较常见，而在肝纤维化中αvβ1占主导作用。整合素αvβ1是一种低亲和力的纤连蛋白受体，在基底上皮细胞中高表达，具有促进角化细胞在底层纤维连接蛋白EDA上迁移的作用。阻断整合素αvβ1与TGF-β1相互作用有助于抑制TGF-β1活性，阻断纤维化进程。此外整合素αvβ1通过激活潜伏的TGFβ1，还可参与牙龈成纤维细胞的合成(Jakhu et al,Journal of oral biology and craniofacial research,2018,8(2):122.)。在众多αv家组蛋白中，针对αvβ6的研究较为深入。有报道称，在正常的肺组织中整合素αvβ6表达量很低，但是当肺损伤发生炎症和纤维化时，αvβ6快速高表达(Hatley et al,Angewandte Chemie International Edition,2018,57(13):3298.)。在患有原发性胆汁性肝硬化(PBC)、酒精性脂肪肝、乙肝、丙肝等疾病的患者中，整合素αVβ6的mRNA表达量升高。在肾疾病相关的慢性炎症和纤维化疾病中整合素αVβ6的表达较于正常肾组织显著提高。此外，在糖尿病、肺出血肾炎综合症、Alport综合症、狼疮性肾炎等病人活检样品中整合素αVβ6都显著高表达(Koivisto et al,The international journal of biochemistry&cell biology,2018,99:186.)。
组织纤维化可发生于多种器官，是一种较为常见的纤维化疾病包括特发性肺纤维化(IPF)、非酒精性脂肪肝(NASH)、肝硬化、肾纤维化、硬皮症、心肌纤维化等。组织损 伤和炎症是纤维化的重要诱因。由于炎症导致器官实质细胞发生坏死，局部免疫细胞被激活，多种血细胞进入损伤部位。被激活的免疫细胞产生大量具有高度生物活性的细胞因子和趋化因子，导致间充质细胞的局部激活，这些细胞产生细胞外基质(ECM)，破坏细胞外微环境，并进一步增加促炎细胞因子、趋化因子和血管生成因子的产生。最终，细胞外基质异常增多和过度沉积而发生病变致使组织纤维化(Ricard-Blum et al,Matrix Biology,2018,68:122.)。纤维化的主要特征是形成和沉积过多的纤维结缔组织。慢性的伴纤维化损伤会使组织结构被破坏、器官功能发生障碍最终导致器官衰竭。
目前获批上市的整合素抑制剂尚为数不多，亟需开发一类结构新颖的，活性好且更加安全有效的整合素调节剂。
发明内容
为解决现有技术中存在的问题，本发明提供一种式I所示的化合物及其消旋体、立体异构体、互变异构体、同位素标记物、氮氧化物、溶剂化物、多晶型物、代谢产物、酯、前药或其药学上可接受的盐：
Figure PCTCN2021078537-appb-000001
其中，所述R 1、R 5各自独立的选自H，OH，卤素，CN，SH，NH 2，COOH，任选被一个、两个或更多个R a取代的如下基团：C 1-C 12脂肪烃基，C 1-C 12脂肪烃基氧基；所述R a选自H，＝O，卤素，OH，CN，SH，NH 2，COOH；每一个R 1可以相同或不同；每一个R 5可以相同或不同；
所述W选自-O-，-NR 2-，-S-；
所述R 2、R 3各自独立的选自H，C 1-C 12脂肪烃基，C 3-12环烷基，C 3-12环烷基-C 1-C 12脂肪烃基，-L 5-Ar，并且R 2、R 3中至少一个选自-L 5-Ar；
所述Ar选自任选被一个、两个或更多个R b取代的如下基团：C 3-12环烷基，3-12元杂环基，C 6-20芳基或5-14元杂芳基；所述R b选自H，＝O，卤素，OH，CN，SH，NH 2，COOH，或任选被一个、两个或更多个R c取代的如下基团：C 1-C 12脂肪烃基，C 1-C 12脂肪烃基氧基，C 1-C 12脂肪烃基-SO 2-，C 1-C 12脂肪烃基-NH-，二(C 1-C 12脂肪烃基)N-，C 3-12环烷基，C 3-12环烷基氧基，C 3-12环烷基-SO 2-，C 3-12环烷基-NH-，C 3-12环烷基-C 1-C 12脂肪烃基氧基，C 3-12环烷基-C 1-C 12脂肪烃基-SO 2-，C 3-12环烷基-C 1-C 12脂肪烃基-NH-，3-12元杂环基，3-12元杂环基氧基，3-12元杂环基-SO 2-，3-12元杂环基-NH-，C 6-20芳基，C 6-20芳基氧基，C 6-20芳基-SO 2-，C 6-20芳基-NH-，5-14元杂芳基，5-14元杂芳基氧基，5-14元杂芳基-SO 2-，5-14元杂芳基-NH-；所述R c选自H，＝O，卤素，OH，CN，SH，NH 2，COOH，C 1-C 12脂肪烃基，C 1-C 12脂肪烃基氧基，C 1-C 12脂肪烃基-SO 2-，C 1-C 12脂肪烃基-NH-，二(C 1-C 12脂肪烃基)N-；
所述R 4独立的选自H，C 1-C 12脂肪烃基；
所述L 1、L 2、L 3、L 4、L 5各自独立的选自-(CH 2) n-C(＝X)-或-C(＝X)-(CH 2) n-，-(CH 2) m-(当m＝0时，代表键)，-(CH 2) n-C(＝O)-NH-，-(CH 2) n-NH-C(＝O)-，-C(＝O)-NH-(CH 2) n-，-NH-C(＝O)- (CH 2) n-，-(CH 2) n-C(＝O)-NH-(CH 2) n-，-(CH 2) n-NH-C(＝O)-(CH 2) n-；所述X选自O，NH；所述n，m各自独立的选自0-6；
根据本发明的实施方案，所述L 1、L 2、L 4各自独立的选自-(CH 2) n-C(＝X)-或-C(＝X)-(CH 2) n-，-(CH 2) m-(当m＝0时，代表键)，所述L 3选自-(CH 2) m-(当m＝0时，代表键)；所述L 5选自-NH-C(＝O)-,-C(＝O)-NH-，-(CH 2) m-(当m＝0时，代表键)；所述X选自O，NH；所述n，m各自独立的选自0，1，2，3，4，5，6；
根据本发明的实施方案，优选的，所述L 1选自-(CH 2) 2-，-(CH 2) 3-，-(CH 2) 4-，-(CH 2) 5-，-CH 2-C(＝O)-，-(CH 2) 2-C(＝O)-，-(CH 2) 3-C(＝O)-，-(CH 2) 4-C(＝O)-；L 2选自键，-CH 2-C(＝O)-，-CH 2-，，-C(＝O)-；L 3选自键，-CH 2-；L 4选自键，-CH 2-；L 5选自键，-CH 2-；
根据本发明的实施方案，所述“卤素”选自F、Cl、Br、I；所述“脂肪烃基”选自烷基、烯基、炔基；
根据本发明的实施方案，优选的，所述R 1、R 5各自独立的选自卤素，C 1-C 6脂肪烃基，例如，选自F、Cl、Br、I，甲基，乙基，正丙基，异丙基，正丁基，异丁基，叔丁基，正戊基，异戊基，新戊基，正己基。
根据本发明的实施方案，优选的，所述R 2选自H，C 1-C 6脂肪烃基或C 3-8环烷基-C 1-C 6脂肪烃基，且R 3选自-L 5-Ar或所述R 2选自-L 5-Ar且R 3选自H，C 1-C 6脂肪烃基或C 3-8环烷基-C 1-C 6脂肪烃基；更优选的，所述R 2或R 3之一选自H，甲基，乙基，正丙基，异丙基，正丁基，异丁基，叔丁基，正戊基，异戊基，新戊基，正己基，环丙基甲基，环丁基甲基，环戊基甲基，环己基甲基。
根据本发明的实施方案，所述Ar选自任选被一个、两个或更多个R b取代的如下基团：C 6-C 14芳基，5-10元杂环基，5-6元杂芳基，优选的，Ar选自任选被一个、两个或更多个R b取代的苯基，萘基，2,3-二氢苯并呋喃基，苯并呋喃基，苯并吡喃基，3,4-二氢-2H-1-苯并吡喃基(色烷基)，2,3-二氢苯并[b][1,4]二噁烷基，吡啶基，嘧啶基，吲唑基(1H-吲唑基，2H-吲唑基)，吲哚基，异吲哚基，喹啉基，异喹啉基，喹唑啉基，苯并噁唑基；
根据本发明的实施方案，优选的，所述R b选自H，卤素，CN，或任选被一个、两个或更多个R c取代的如下基团：C 1-C 6脂肪烃基，C 1-C 6脂肪烃基氧基，C 1-C 6脂肪烃基-SO 2-，C 1-C 6脂肪烃基-NH-，二(C 1-C 6脂肪烃基)N-，C 6-10芳基，C 6-10芳基氧基，C 6-10芳基-SO 2-，C 6-10芳基-NH-，5-6元杂芳基，5-6元杂芳基氧基，5-6元杂芳基-SO 2-，5-6元杂芳基-NH-，5-6元杂环基，5-6元杂环基氧基，5-6元杂环基-SO 2-，5-6元杂环基-NH-，C 3-8环烷基，C 3-8环烷基氧基，C 3-8环烷基-SO 2-，C 3-8环烷基-NH-，C 3-8环烷基-C 1-C 6脂肪烃基氧基，C 3-8环烷基-C 1-C 6脂肪烃基-SO 2-，C 3-8环烷基-C 1-C 6脂肪烃基-NH-；更优选的，所述R b选自H，卤素，CN或任选被一个、两个或更多个R c取代的C 1-C 6烷基，C 1-C 6烷氧基，C 1-C 6烷基-SO 2-，C 1-C 6烷基-NH-，二(C 1-C 6烷基)N-，5-6元杂芳基-SO 2-，5-6元杂芳基-NH-，5-6元杂环基，5-6元杂环基氧基，5-6元杂环基-SO 2-，5-6元杂环基-NH-，C 3-6环烷基，C 3-6环烷基氧基，C 3-6环烷基-SO 2-，C 3-6环烷基-NH-，C 3-6环烷基-C 1-C 6烷氧基，C 3-6环烷基-C 1-C 6烷基-SO 2-，C 3-6环烷基-C 1-C 6烷基-NH-；进一步优选的，所述R b选自H，卤素，CN或任选被一个、两个或更多个R c取代的甲基，乙基，正丙基，异丙基，正丁基，异丁基，叔丁基，正戊基，异戊基，新戊基，正己基，乙烯基，1-丙烯基，2-丙烯基，1-甲基乙烯基，1-丁烯基，1-乙基乙烯基，1-甲基-2-丙烯基，2-丁烯基，3-丁烯基、2-甲基-1-丙烯基，2-甲基-2-丙烯基，1-戊烯基、1- 己烯基，乙炔基，1-丙炔基，2-丙炔基，1-丁炔基，1-甲基-2-丙炔基，3-丁炔基，1-戊炔基，1-己炔基，甲氧基，乙氧基，正丙氧基，异丙氧基，异丁氧基，正丁氧基，叔丁氧基，戊氧基，己氧基，N,N-二甲基氨基，N,N-二乙基氨基，甲基-NH-，乙基-NH-，正丙基-NH-，异丙基-NH-，正丁基-NH-，异丁基-NH-，叔丁基-NH-，正戊基-NH-，异戊基-NH-，新戊基-NH-，正己基-NH-，甲基-SO 2-，乙基-SO 2-，正丙基-SO 2-，异丙基-SO 2-，正丁基-SO 2-，异丁基-SO 2-，叔丁基-SO 2-，正戊基-SO 2-，异戊基-SO 2-，新戊基-SO 2-，正己基-SO 2-，环丙基，环丁基，环戊基，环己基，环丙基氧基，环丁基氧基，环戊基氧基，环己基氧基，环丙基，环丁基，环戊基，环己基，环丙基-SO 2-，环丁基-SO 2-，环戊基-SO 2-，环己基-SO 2-，环丙基-NH-，环丁基-NH-，环戊基-NH-，环己基-NH-，环丙基甲基氧基，环丁基甲基氧基，环戊基甲基氧基，环己基甲基氧基，环丙基甲基-SO 2-，环丁基甲基-SO 2-，环戊基甲基-SO 2-，环己基甲基-SO 2-，环丙基甲基-NH-，环丁基甲基-NH-，环戊基甲基-NH-，环己基甲基-NH-，苯基，苯基-SO 2-，苯基-NH-，萘基，萘基-SO 2-，萘基-NH，氧杂环丁烷，吡喃基，四氢吡喃基，呋喃基，四氢呋喃基，四氢吡咯基，哌啶基，吡啶基，吡嗪基，吡咯基，咪唑基，吡唑基，三氮唑，噁唑基，异噁唑基，吗啉基；例如，R b选自F，Cl，Br，I，CN，甲基，乙基，正丙基，异丙基，正丁基，异丁基，叔丁基，新戊基，甲基-SO 2-，甲氧基，乙氧基，正丙氧基，异丙氧基，异丁氧基，N,N-二甲基氨基，异丙基-NH-，环丙基氧基，环丁基氧基，环戊基氧基，环丙基甲基氧基，环丙基氨基，CF 3，CHF 2，CH 2F，CF 3O，CHF 2O，CH 2FO，苯基，3,5-二甲基吡唑-1-基，咪唑基，吗啉基，1,2,4-三氮唑，氧杂环丁烷，四氢呋喃基，四氢吡喃基，四氢吡咯基；
根据本发明的实施方案，优选的，所述R c选自H，＝O，卤素，OH，CN，SH，NH 2，COOH，C 1-C 6烷基，C 1-C 6烷氧基，C 1-C 6烷基-SO 2-，二(C 1-C 6烷基)N-，C 1-C 6烷基-NH-；例如选自H，＝O，F，Cl，Br，I，OH，CN，SH，NH 2，COOH，甲基，乙基，正丙基，异丙基，正丁基，异丁基，叔丁基，正戊基，异戊基，新戊基，正己基，甲氧基，乙氧基，正丙氧基，异丙氧基，异丁氧基，正丁氧基，叔丁氧基，戊氧基，己氧基，N,N-二甲基氨基，N,N-二乙基氨基，甲基-NH-，乙基-NH-，正丙基-NH-，异丙基-NH-，正丁基-NH-，异丁基-NH-，叔丁基-NH-，正戊基-NH-，异戊基-NH-，新戊基-NH-，正己基-NH-，甲基-SO 2-，乙基-SO 2-，正丙基-SO 2-，异丙基-SO 2-，正丁基-SO 2-，异丁基-SO 2-，叔丁基-SO 2-，正戊基-SO 2-，异戊基-SO 2-，新戊基-SO 2-，正己基-SO 2-；
根据本发明的实施方案，所述R 4独立的选自H，C 1-C 6脂肪烃基，优选的，选自甲基，乙基，正丙基，异丙基，正丁基，异丁基，叔丁基，正戊基，异戊基，新戊基，正己基；
根据本发明的实施方案，优选的，所述Ar选自如下基团：
Figure PCTCN2021078537-appb-000002
Figure PCTCN2021078537-appb-000003
所述结构中，L 5(不为键)，R b，R c选自式I中所述定义。
根据本发明的实施方案，更优选的，所述R 2或R 3中的一个选自如下基团：
Figure PCTCN2021078537-appb-000004
Figure PCTCN2021078537-appb-000005
Figure PCTCN2021078537-appb-000006
根据本发明的实施方案，所述式I结构中存在多个手性中心，例如包括如下手性中心(如下式所示，分别标记为*S 1、*S 2)：
Figure PCTCN2021078537-appb-000007
其中，所述*S 1处的构型可选自：
Figure PCTCN2021078537-appb-000008
所述*S 2处的构型可选自：
Figure PCTCN2021078537-appb-000009
根据本发明的实施方案，所述式I可以选自包括一个手性中心的特定立体构型，或两个以上手性中心不同特定立体构型的组合的化合物结构。
根据本发明的实施方案，所述式I结构可以选自式II：
Figure PCTCN2021078537-appb-000010
式II所述式II结构中，R 1、R 2、R 3、R 4、R 5、L 1、L 2、L 3、L 4以及其他手性中心如上前述式I所定义。例如，所述式II结构进一步如下式IIa-IIt所示：
Figure PCTCN2021078537-appb-000011
Figure PCTCN2021078537-appb-000012
Figure PCTCN2021078537-appb-000013
Figure PCTCN2021078537-appb-000014
根据本发明的实施方案，所述式I化合物进一步优选为如下式III(式IIIa、式IIIb)、式IV(式IVa、式IVb)：
Figure PCTCN2021078537-appb-000015
Figure PCTCN2021078537-appb-000016
根据本发明的实施方案，所述式III(式IIIa、式IIIb)、式IV(式IVa、式IVb)结构中，R 1、R 3、R 4、R 5、L 1、L 3以及其他手性中心如上前述式I所定义。
根据本发明的实施方案，所述式I化合物进一步优选为如下式V至式XVII：
Figure PCTCN2021078537-appb-000017
Figure PCTCN2021078537-appb-000018
Figure PCTCN2021078537-appb-000019
根据本发明的实施方案，所述式V至式XVII结构中，R 1、R 2、R 3、R 4、R 5、n、m以及其他手性中心如上前述式I所定义。
根据本发明的实施方案，所述式I(包括式II至式XVII)所示的化合物及其消旋体、立体异构体、互变异构体、同位素标记物、氮氧化物、溶剂化物、多晶型物、代谢产物、酯、前药或其药学上可接受的盐中，式I化合物的举例性的、非限制性的具体实例如下所示：
Figure PCTCN2021078537-appb-000020
Figure PCTCN2021078537-appb-000021
Figure PCTCN2021078537-appb-000022
Figure PCTCN2021078537-appb-000023
Figure PCTCN2021078537-appb-000024
Figure PCTCN2021078537-appb-000025
Figure PCTCN2021078537-appb-000026
Figure PCTCN2021078537-appb-000027
Figure PCTCN2021078537-appb-000028
所述结构式中，以星号(“*”)标记表明所述结构在星号所示碳位具有手性，因而可以以所述标记位置具有相反构象的异构体混合物形式存在，并通过手性拆分得到具有所述结构的两种异构体(参见实施例3，例如化合物5的差向异构体混合物形式可以通过手性拆分得到星号位置构型不同的化合物5a和5b)。根据本发明的实施方案，所述式I化合物的立体异构体还可以进一步选自例如如下结构：
Figure PCTCN2021078537-appb-000029
Figure PCTCN2021078537-appb-000030
本发明还提供所述式I所示的化合物(包括式II至式XVII)及其消旋体、立体异构体、互变异构体、同位素标记物、氮氧化物、溶剂化物、多晶型物、代谢产物、酯、前药或其药学上可接受的盐的制备方法，但不仅限于以下描述的方法。所有的原料都是根据符合通式规律的目标分子的基团特征，并通过这些路线中的方案、有机化学领域普通技术人员熟知的方法制备或者直接购买的。可将用下述方法和合成有机化学领域中已知的合成方法或本领域技术人员意识到的有关改变方法结合在一起，合成本发明化合物。本领域技术人员可知，根据特定的目标结构，可以任选采用下述一种或几种方案进行结合，或者一种或几种方案中的任意步骤进行组合得到合成方案。
本发明所述化合物的制备方法包括：在合适的条件下，将含萘啶环结构的原料与含吡咯 烷结构的原料在合适的试剂中进行反应，并任选的，在合适的条件下，进行上保护基，脱保护基，取代，缩合，还原胺化或水解步骤。具体的，可以参照如下方案进行合成。
在第一种方案中，本发明的化合物的制备包括如下步骤中的一步或几步：
Figure PCTCN2021078537-appb-000031
所述方案中，所述R 1，R 3，R 4(R 4≠H)，R 5，L 1，L 2，L 3，L 4如前述式I所定义；所述Lx选自L 1-X 1，其中，X 1为离去基团，选自例如OTs，OMs，OTf，卤素(Cl，Br，I)，或者所述Lx选自L 1基团(当存在末端-CH 2-时)的末端-CH 2-替代为CHO或COOH的基团，例如选自-(CH 2) m-1-CHO或-(CH 2) m-1-COOH，所述m如前述式I所定义；所述PG 1为N上保护基，选自Boc-等；所述第一种方案中，采用化合物A-1和A-2合成得到化合物A-3，反应条件可选自(i)NaBH(OAc) 3，NaBH 3CN，HOAc,甲醇；或(ii)HBTU，TEA，DMF和或任选的加入酸性试剂；或(iii)DIPEA，乙腈；(iv)K 2CO 3，NaI，乙腈；所述脱保护基条件包括在酸性试剂，第一种有机溶剂下进行，所述酸性试剂选自三氟乙酸(TFA)，盐酸，所述第一种有机溶剂选自二氯甲烷，1，4-二氧六环，醇类试剂中的一种或多种，所述醇类试剂选自甲醇，乙醇；所述水解条件包括在碱性条件，水+第二种有机溶剂条件下进行，所述碱性条件选自碱金属或碱土金属的氢氧化物，例如选自LiOH，NaOH，所述第二种有机溶剂选自醇类试剂，例如选自甲醇、乙醇。
根据第一种方案，其可以进一步选自如下方案1a：
Figure PCTCN2021078537-appb-000032
所述方案中，所述R 2，R 3，R 4，m如前述式I所定义，其中m不为0；所述X 1，PG 1如前定义；
根据第一种方案，其可以进一步选自如下方案1b：
Figure PCTCN2021078537-appb-000033
所述方案中，所述R 2，R 3，R 4(R 4≠H)，n如前述式I所定义；所述X 1，PG 1如前定义；
根据第一种方案，其可以进一步选自如下方案1c：
Figure PCTCN2021078537-appb-000034
所述方案中，所述R 2，R 3，R 4(R 4≠H)，n如前述式I所定义；所述PG 1如前定义；
根据第一种方案，其可以进一步选自如下方案1d：
Figure PCTCN2021078537-appb-000035
所述方案中，所述R 2，R 3，R 4(R 4≠H)，m如前述式I所定义；所述PG 1如前定义；
根据第一种方案，其可以进一步选自如下方案1e：
Figure PCTCN2021078537-appb-000036
所述方案中，所述R 2，R 3，R 4(R 4≠H)，m如前述式I所定义；所述PG 1如前定义；
上述合成方案中，若需要合成含有
Figure PCTCN2021078537-appb-000037
特定立体构型的反应产物，可以替代原料为相应的含有
Figure PCTCN2021078537-appb-000038
的构型；
在第二种方案中，本发明的化合物的制备还包括如下步骤中的一步或几步；
Figure PCTCN2021078537-appb-000039
所述方案中，所述R 3，R 4，R 5，L 4如前述式I所定义；所述PG 1如前定义；所述化合物B-1和B-2在还原剂，酸，有机溶剂存在条件下得到化合物B-3，所述还原剂选自NaBH 3CN，所述酸选自乙酸，所述有机溶剂选自醇类试剂，例如甲醇，乙醇；所述化合物B-3和化合物B-5分别在酸性条件下，有机溶剂中水解得到B-4和B-6，所述酸性条件选自TFA，所述有机溶剂选自DCM；所述B-3经过NH甲基化反应得到化合物B-5，所述甲基化反应条件为多聚甲醛，NaBH(OAC) 3，NaBH 3CN，有机溶剂中进行，所述有机溶剂选自醇类试剂，例如甲醇，乙醇。
在第三种方案中，本发明的化合物的制备包括如下步骤：
Figure PCTCN2021078537-appb-000040
根据本发明的实施方案，化合物C-1可拆分为化合物C-2，C-3，所采用的拆分条件包括使用手性色谱柱，所述方案中，所述R 1，R 2，R 3，R 4，R 5，L 1，L 3如前述式I所定义。
在第四种方案中，本发明的化合物的制备包括如下步骤中的一步或几步；
Figure PCTCN2021078537-appb-000041
Figure PCTCN2021078537-appb-000042
所述方案中，X 2为离去基团，选自卤素，例如Cl，Br，I；所述R 1，R 2，R 3，R 4，R 5如前述式I所定义，所述PG 1同前述定义。所述化合物D-1在碱性试剂、催化剂和有机试剂存在条件下进行反应得到化合物D-2，所述碱性试剂选自碱金属或碱土金属的碳酸盐，例如为K 2CO 3，Na 2CO 3，所述催化剂为碘化物，例如为NaI，所述有机试剂选自乙腈，DMF，DMSO；所述化合物D-2经过NH甲基化反应得到化合物D-3，所述甲基化反应条件为多聚甲醛，NaBH(OAC) 3，NaBH 3CN，有机溶剂中进行，所述有机溶剂选自醇类试剂，例如甲醇，乙醇；所述化合物D-4在H 2SO 4，烷基醇试剂存在下得到化合物D-5，所述烷基醇试剂可选自甲醇；所述化合物D-5在卤代试剂，有机溶剂，引发剂存在条件下反应得到化合物D-1，所述卤代试剂选自NBS，CBr 4，所述引发剂选自AIBN，所述有机溶剂选自四氯化碳，二氯甲烷，三氯甲烷，1，4-二氧六环，四氢呋喃；所述D-1和D-6在碱性试剂，有机溶剂存在条件下生成D-7，所述碱性试剂选自碱金属或碱土金属的碳酸盐，所述有机溶剂选自二氯甲烷，三氯甲烷，四氯化碳。
根据第四种方案，其可以进一步选自如下方案4a：
Figure PCTCN2021078537-appb-000043
上述合成方案中，可以替代
Figure PCTCN2021078537-appb-000044
为
Figure PCTCN2021078537-appb-000045
得到不同构型的产物；
在第五种方案中，本发明的化合物的制备包括如下步骤中的一步或几步：
Figure PCTCN2021078537-appb-000046
所述方案中，所述R 2，R 4，R 5如前述式I所定义，且所述R 2选自-L 5-Ar；所述PG 1同前述定义；所述化合物E-1与有机胺试剂反应得到E-2，所述反应在还原试剂，酸性试剂，有机溶剂存在条件下进行，所述还原试剂选自BH 3·THF，所述酸性试剂选自乙酸，所述有机溶剂选自THF；所述化合物E-2与E-3反应得到E-4，所述反应在碱性试剂，催化剂，有机溶剂存在条件下进行，所述碱性试剂选自碱金属或碱土金属的碳酸盐，例如为K 2CO 3，Na 2CO 3，所述催化剂为碘化物，例如为NaI，所述有机试剂选自乙腈，DMF，DMSO。
在第六种方案中，本发明的化合物的制备包括如下步骤：
Figure PCTCN2021078537-appb-000047
所述方案中，所述p，q，R b，R 4，R 5如前述式I所定义；所述PG 1同前述定义；所述化合物F-1与F-2反应得到F-3，所述反应在还原试剂，酸性试剂，有机溶剂存在条件下进行，所述还原试剂选自三乙酰氧基硼氢化钠，氰基硼氢化钠，所述酸性试剂选自乙酸，所述有机溶剂选自醇类试剂，例如甲醇，乙醇。
根据第六种方案，其可以进一步选自如下方案6-a：
Figure PCTCN2021078537-appb-000048
在第七种方案中，本发明的化合物的制备包括如下步骤中的一步或几步：
Figure PCTCN2021078537-appb-000049
所述方案中，所述R 4如前述式I所定义，所述X 3选自卤素，例如F，Cl，Br，I；所述化合物G-1与卤代烷烃试剂反应生产化合物G-2，反应在碱性试剂，有机溶剂条件下进行，所述碱性试剂选自碱金属或碱土金属的碳酸盐，例如为K 2CO 3，Na 2CO 3，所述有机试剂选自乙腈，DMF，DMSO；所述化合物G-2与G-3反应产生G-4，反应在碱性试剂，有机溶剂条件下进行，所述碱性试剂选自NaH，LiH，KH，所述有机溶剂选自甲苯，苯。
在第八种方案中，本发明的化合物的制备包括如下步骤：
Figure PCTCN2021078537-appb-000050
所述方案中，所述R b如前述式I所定义，所述化合物H-1(盐酸盐)反应得到H-2和H-3，反应在酸性试剂，水，有机溶剂存在条件下进行，所述酸性试剂选自盐酸，所述有机溶剂选自醇类试剂，例如甲醇，乙醇，所述反应优选在微波条件下进行。
在第九种方案中，本发明的化合物的制备包括如下步骤中的一步或几步：
Figure PCTCN2021078537-appb-000051
所述方案中，所述R 4、R b如前述式I所定义，所述X 4选自卤素，例如F，Cl，Br，I；所述化合物I-1反应生成I-2，反应在有机溶剂，Pb(OAc) 2，碱性试剂存在条件下进行，所述碱性试剂选自三乙胺，吡啶等，所述有机溶剂选自1，4-二氧六环。所述I-2在碱性试剂存在条件下生成I-3，所述碱性剂选自NaH，KH，LiH。
在第十种方案中，本发明的化合物的制备包括如下步骤：
Figure PCTCN2021078537-appb-000052
所述方案中，所述R 4、R b如前述式I所定义，所述化合物J-1和J-2反应生成J-3，反应在二氯亚砜，氯化镁，有机溶剂存在条件下进行，所述有机溶剂选自四氢呋喃，二氯甲烷，三氯甲烷。
在第十一种方案中，本发明的化合物的制备包括如下步骤中的一步或几步：
Figure PCTCN2021078537-appb-000053
Figure PCTCN2021078537-appb-000054
在第十二种方案中，本发明的化合物的制备包括如下步骤：
Figure PCTCN2021078537-appb-000055
所述方案中，R 1，m如前述式I所定义，所述PG 1同前述定义；所述化合物L-1水解为L-2，反应在碱性试剂，有机溶剂/水反应条件下进行，所述碱性试剂选自碱金属或碱土金属的氢氧化物，例如选自NaOH，KOH，LiOH，所述有机溶剂选自醇类试剂，例如选自甲醇，乙醇；所述化合物L-3反应得到L-4，在还原试剂、有机溶剂存在条件下反应，所述还原试剂选自DIBAL-H，所述有机溶剂选自THF，二氯甲烷，三氯甲烷等。
在第十三种方案中，本发明的化合物的制备包括如下步骤中的一步或几步：
Figure PCTCN2021078537-appb-000056
所述方案中，所述R b，R 3，R 4，R 5如前述式I所定义，所述PG 1同前述定义。
在第十四种方案中，本发明的化合物的制备包括如下步骤：
Figure PCTCN2021078537-appb-000057
在第十五种方案中，本发明的化合物的制备包括如下步骤中的一步或几步：
Figure PCTCN2021078537-appb-000058
所述方案中，所述R 4，R 5如前述式I所定义，所述PG 1同前述定义，所述X 5选自卤素，例如F，Cl，Br，I。
在第十六种方案中，本发明的化合物的制备包括如下步骤中的一步或几步：
Figure PCTCN2021078537-appb-000059
所述方案中，R 4如前述式I所定义，所述PG 1同前述定义，所述X 6选自卤素，例如F，Cl，Br，I。
在第十六种方案中，本发明的化合物的制备包括如下步骤中的一步或几步：
Figure PCTCN2021078537-appb-000060
所述方案中，R b选自如前式I定义中的C 1-C 12烷氧基，C 3-C 8环烷基氧基或3-12元杂环基氧基。所述化合物P-1在烷基醇，酸性试剂存在条件下得到化合物P-2，所述烷基醇选自甲醇，乙醇，所述酸性试剂选自硫酸；化合物P-2在卤代脂肪烃或卤代脂环烃试剂，有机溶剂，碱性试剂存在条件下反应得到化合物P-3，所述卤代脂肪烃或卤代脂环烃试剂可以选自卤代(C 1-C 12)烷烃，卤代(C 3-C 8)环烷烃或卤代3-12元杂环，例如选自2-碘代丙烷，3-碘氧杂环丁烷；所述碱性试剂选自碱金属或碱土金属的碳酸盐，例如碳酸钾，碳酸钠，碳酸铯，所述有机溶剂选自DMF，DMSO，乙腈。
在第十七种方案中，本发明的化合物的制备包括如下步骤中的一步或几步：
Figure PCTCN2021078537-appb-000061
所述方案中，X 7为卤素，选自F，Cl，Br，I。
在第十八种方案中，本发明的化合物的制备包括如下步骤：
Figure PCTCN2021078537-appb-000062
所述方案中，R 4如前述式I所定义，X 8为卤素，选自F，Cl，Br，I。
在第十九种方案中，本发明的化合物的制备包括如下步骤：
Figure PCTCN2021078537-appb-000063
所述方案中，R b如前述式I所定义。
在第二十种方案中，本发明的化合物的制备包括如下步骤：
Figure PCTCN2021078537-appb-000064
所述方案中，R b如前述式I所定义，X 9为卤素，选自F，Cl，Br，I，所述反应条件为碱性试剂，有机溶剂以及烷基醇试剂存在下，所述碱性试剂选自碱金属或碱土金属的氢氧化物，例如KOH，NaOH，所述有机溶剂选自DMSO，DMF，乙腈，所述烷基醇试剂选自甲醇，乙醇等。
在第二十一种方案中，本发明的化合物的制备包括如下步骤：
Figure PCTCN2021078537-appb-000065
所述方案中，R 4如前述式I所定义，X 10为卤素，选自F，Cl，Br，I。
本发明进一步提供一种药物组合物，其包含本发明所述的式I化合物及其消旋体、立体异构体、互变异构体、同位素标记物、氮氧化物、溶剂化物、多晶型物、代谢产物、酯、前药或其药学上可接受的盐。
在一些实施方案中，本发明所述的药物组合物进一步包含治疗有效量的本发明所述式I化合物及其消旋体、立体异构体、互变异构体、同位素标记物、氮氧化物、溶剂化物、多晶型物、代谢产物、酯、前药或其药学上可接受的盐和药学上可接受的载体。
所述药物组合物中的载体为“可接受的”，其可与组合物的活性成分相容(并且优选地，能够稳定活性成分)并且对被治疗的受试者不是有害的。可以使用一种或多种增溶剂作为药物赋形剂用于递送活性化合物。
本发明进一步提供所述式I化合物及其消旋体、立体异构体、互变异构体、同位素标记物、氮氧化物、溶剂化物、多晶型物、代谢产物、酯、前药或其药学上可接受的盐或所述药物组合物在制备整合素调节剂中的应用。
本发明进一步提供所述式I化合物及其消旋体、立体异构体、互变异构体、同位素标记物、氮氧化物、溶剂化物、多晶型物、代谢产物、酯、前药或其药学上可接受的盐或所述药物组合物在制备预防，调节或治疗与整合素活性有关的疾病或病症的药物中的用途。
本发明进一步提供所述式I化合物及其消旋体、立体异构体、互变异构体、同位素标记物、 氮氧化物、溶剂化物、多晶型物、代谢产物、酯、前药或其药学上可接受的盐或所述药物组合物在制备用于治疗纤维化性疾病，炎性疾病或细胞增殖性疾病的药物中的用途。
本发明进一步提供所述式I化合物及其消旋体、立体异构体、互变异构体、同位素标记物、氮氧化物、溶剂化物、多晶型物、代谢产物、酯、前药或其药学上可接受的盐或所述药物组合物在制备用于抑制细胞中TGF-β活化的药物中的用途。
本发明还提供了一种在受试者中调节至少一种整合素活性的方法，该方法包括给予本发明所述化合物作为治疗剂。
本发明中，化合物对整合素的调节作用是对αVβ1，αVβ3，αVβ5，αVβ6，和αVβ8，α5β1，α8β1中的任一种或αVβ1，αVβ3，αVβ5，αVβ6，和αVβ8，α5β1，α8β1中的一个或多个的组合的调节作用。在一些实施方式中，所述调节作用表现为抑制作用。在一些实施方式中，所述抑制作用可以是对αVβ1，αVβ3，αVβ5，αVβ6，和αVβ8，α5β1，α8β1的抑制作用；在另一些实施方式中，所述抑制作用是对αVβ1，αVβ3，αVβ5，αVβ6，和αVβ8，α5β1，α8β1中的一种；在另一些实施方式中，所述抑制作用包括对α8β1和αVβ1的抑制作用；在另一些实施方式中，所述抑制作用包括对α8β1和α5β1的抑制作用；在另一些实施方式中，所述抑制作用包括对αvβ3和αvβ5的抑制作用；在一些实施方式中，所述抑制作用包括对α8β1，αvβ3和αvβ5的抑制作用；在一些实施方式中，所述抑制作用包括对α8β1，αvβ1和α5β1的抑制作用；在一些实施方式中，所述抑制作用包括对α8β1，αvβ1，αvβ3和αvβ5的抑制作用。根据本发明的实施方案，所述整合素包括αVβ1，αVβ3，αVβ5，αVβ6，和αVβ8，α5β1，α8β1中的一个或多个的组合。本发明还提供了一种用于治疗疾病或病症的方法，该方法包括向需要这种治疗的患者单独施用治疗有效量的至少一种本发明的化合物，或任选地，与本发明的另一种化合物和/或至少一种其他类型的治疗剂组合。
本发明还提供了一种抑制细胞中TGF-β活化的方法，该方法包括向该细胞施用式I化合物及其消旋体、立体异构体、互变异构体、同位素标记物、氮氧化物、溶剂化物、多晶型物、代谢产物、酯、前药或其药学上可接受的盐或所述药物组合物。
在一些实施方案中，所述疾病或病症与纤维化有关，包括肺，肝，肾，心脏，皮肤，眼和胰腺纤维化。
根据本发明的实施方案，所述疾病或病症与细胞增殖性病症例如癌症有关。在一些实施方案中，癌症包括实体瘤生长或瘤形成。在其他实施方案中，癌症包括肿瘤转移。在一些实施方案中，癌症是膀胱癌，血液癌，骨癌，脑癌，乳腺癌，中枢神经系统癌，子宫颈癌，结肠癌，子宫内膜癌，食道癌，胆囊癌，生殖器癌，泌尿生殖道癌，头癌，肾癌，喉癌，肝癌，肺癌，肌肉癌。组织，颈部，口腔或鼻粘膜，卵巢，胰腺，前列腺，皮肤，脾脏，小肠，大肠，胃，睾丸或甲状腺。在其他实施方案中，癌症是肉瘤，淋巴瘤，白血病，黑素瘤，间皮瘤，多发性骨髓瘤或精原细胞瘤。
根据本发明可以预防，调节或治疗的与αV整合素活性有关的疾病，病症或病状的实例包括但不限于移植注射，纤维化病症(例如特发性肺纤维化，间质性肺疾病，肝纤维化，非酒精性脂肪性肝，原发性硬化性胆管炎(PSC)，肾纤维化，皮肤纤维化，心肌纤维化，系统性硬化)，炎症性疾病(例如急性肝炎，慢性肝炎，，牛皮癣，肠易激综合征(IBS)，炎症性肠病(IBD))，骨质疏松症以及细胞增殖性疾病(例如癌症，骨髓瘤，纤维瘤，肝癌，白血病，卡波西氏肉瘤，实体瘤)。
适用于通过本发明化合物预防或治疗的纤维化疾病，炎性疾病以及细胞增殖性疾病包括但不限于特发性肺纤维化(IPF)，间质性肺病，非特异性间质性肺炎(NSIP)，常规间质性肺炎(UIP)，辐射诱发性纤维化，家族性肺纤维化，气道纤维化，慢性阻塞性肺疾病(COPD)，糖尿病性肾病，局灶性节段性肾小球硬化，IgA肾病，药物或移植引起的肾病，自身免疫性肾病，狼疮性肾炎，肝纤维化，肾脏纤维化，慢性肾脏病(CKD)，糖尿病肾病(DKD)，皮肤纤维化，瘢痕，全身性硬化，硬皮病，病毒性纤维化，非酒精性脂肪肝病(NAFLD)，酒精性或非酒精性脂肪性肝炎(NASH)，急性肝炎，慢性肝炎，肝硬化，原发性硬化性胆管炎，药物性肝炎，胆汁性肝硬化，门脉高压，再生衰竭，肝功能不全，肝血流异常，肾病，肺炎，牛皮癣，肠易激综合征罗马(IBS)，炎性肠病(IBD)，胰腺分泌异常，前列腺增生，神经性膀胱疾病，脊髓肿瘤，椎间盘疝，椎管狭窄，心力衰竭，心脏纤维化，血管纤维化，血管周纤维化，足口疾病，癌症，骨髓瘤，纤维瘤，肝癌，白血病，慢性淋巴细胞性白血病，卡波济肉瘤，实体瘤，脑梗死，脑出血，神经性疼痛，周围神经病，年龄相关性黄斑变性(AMD)，青光眼，眼纤维化，角膜瘢痕形成，糖尿病性视网膜病变，增生性玻璃体视网膜病变(PVR)，瘢痕性天疱疮性青光眼滤过手术瘢痕，克罗恩病或系统性红斑狼疮；伤口愈合异常导致瘢痕形成；器官移植后发生纤维化，骨髓纤维化和肌瘤。本发明还提供了一种用于治疗纤维化疾病，炎性疾病或细胞增殖性疾病的方法，包括向需要这种治疗的患者施用治疗有效量的至少一种化合物。单独地或任选地与本发明的另一种化合物和/或至少一种其他类型的治疗剂组合的本发明的“抗氧化剂”。在另一些实施方案中，本发明提供了用于治疗的本发明化合物。
本发明的化合物可以与另外的治疗剂例如一种或多种抗纤维化和/或抗炎治疗剂组合使用。
本发明进一步提供了一种用于治疗纤维化疾病，炎性疾病或细胞增殖性疾病的方法，所述方法包括向有需要的患者施用治疗有效量的第一和第二治疗剂，其中第一治疗剂是本发明的化合物。在一些实施方案中，本发明提供了本发明所述化合物与另外的治疗剂的组合制剂，用于在治疗中同时，分开或依次使用。
术语解释：
除非另有说明，本申请说明书和权利要求书中记载的基团和术语定义，包括其作为实例的定义、示例性的定义、优选的定义、表格中记载的定义、实施例中具体化合物的定义等，可以彼此之间任意组合和结合。这样的组合和结合后的基团定义及化合物结构，应当属于本申请说明书记载的范围。
本申请说明书和权利要求书记载的数值范围，当该数值范围被定义为“整数”时或按照本领域常规理解通常为“整数”时，应当理解为记载了该范围的两个端点以及该范围内的每一个整数。例如，“0～6”表示碳数时，应当理解为记载了0、1、2、3、4、5和6的每一个整数。“更多个”表示三个或三个以上。
术语“卤素”指F、Cl、Br和I。换言之，F、Cl、Br和I在本说明书中可描述为“卤素”。
本文所述任选的被取代基所取代的情形涵盖了无取代以及被一个或多个取代基所取代的情形，例如“任选被一个、两个或更多个R取代”意味着可以不被R取代(无取代)或被一个、两个或更多个R取代。
术语“脂肪烃基”包括饱和或不饱和，直链或支链的链状或环状烃基，所述脂肪烃基的类 型可选自烷基、烯基、炔基等，所述脂肪烃基的碳原子数优选为1-12，还可以为1-10，进一步的优选范围为1-6，具体可包括但不限于如下基团：甲基、乙基、正丙基、异丙基、正丁基、异丁基、叔丁基、正戊基、异戊基、新戊基、正己基、乙烯基、1-丙烯基、2-丙烯基、1-甲基乙烯基、1-丁烯基、1-乙基乙烯基、1-甲基-2-丙烯基、2-丁烯基、3-丁烯基、2-甲基-1-丙烯基、2-甲基-2-丙烯基、1-戊烯基、1-己烯基、乙炔基，1-丙炔基，2-丙炔基，1-丁炔基，1-甲基-2-丙炔基，3-丁炔基，1-戊炔基、1-己炔基、环丙基、环丁基、环戊基和环己基；
术语“C 3-12环烷基”应理解为表示饱和或不饱和的一价单环或双环，其具有3-12个碳原子，优选C 3-8环烷基，更优选的为C 3-6环烷基。例如C 3-8环烷基应理解为表示饱和或不饱和的一价单环或双环，其具有3、4、5、6、7或8个碳原子。所述C 3-12环烷基可以是单环烃基，如环丙基、环丁基、环戊基、环己基、环庚基、环辛基、环壬基或环癸基，或者是双环烃基如四氢化萘或十氢化萘。
术语“3-12元杂环基”意指饱和或不饱和的一价单环或双环，其包含1-5个独立选自N、O和S的杂原子，含杂原子的基团不具有芳香性，所述3-12元杂环基，优选3-10元杂环基。术语3-12元杂环基意指饱和的一价单环或双环，其包含1-5个，优选1-3个选自N、O和S的杂原子。所述杂环基可以通过所述碳原子中的任一个或氮原子(如果存在的话)与分子的其余部分连接。特别地，所述杂环基可以包括但不限于：4元环，如氮杂环丁烷基、氧杂环丁烷基；5元环，如四氢呋喃基、四氢噻吩基、二氧杂环戊烯基、吡咯烷基、咪唑烷基、吡唑烷基、吡咯啉基；或6元环，如四氢吡喃基、哌啶基、吗啉基、二噻烷基、硫代吗啉基、哌嗪基或三噻烷基；或7元环，如二氮杂环庚烷基。任选地，所述杂环基可以是苯并稠合的。所述杂环基可以是双环的，例如但不限于5,5元环，如六氢环戊并[c]吡咯-2(1H)-基环，或者5,6元双环，如六氢吡咯并[1,2-a]吡嗪-2(1H)-基环。含氮原子的环可以是部分不饱和的，即它可以包含一个或多个双键，例如但不限于2,5-二氢-1H-吡咯基、4H-[1,3,4]噻二嗪基、4,5-二氢噁唑基或4H-[1,4]噻嗪基，或者，它可以是苯并稠合的，例如但不限于二氢异喹啉基，2,3-二氢苯并呋喃基，3,4-二氢-2H-1-苯并吡喃基(色烷基)，2,3-二氢苯并[b][1,4]二噁烷基。所述3-12元杂环基还可以选自例如如下基团：
Figure PCTCN2021078537-appb-000066
术语“C 6-20芳基”应理解为优选表示具有6-20个碳原子的一价芳香性或部分芳香性的单环、双环或三环烃环，优选“C 6-10芳基”。术语C 6-20芳基应理解为优选表示具有6、7、8、9、10、11、12、13、14、15、16、17、18、19、20个碳原子的一价芳香性或部分芳香性的单环、双环或三环烃环，特别是具有6个碳原子的环(“C 6芳基”)，例如苯基；或联苯基，或者是具有9个碳原子的环(“C 9芳基”)，例如茚满基或茚基，或者是具有10个碳原子的环(“C 10芳基”)，例如四氢化萘基、二氢萘基或萘基，或者是具有13个碳原子的环(“C 13芳基”)，例如芴基，或者是具有14个碳原子的环(“C 14芳基”)，例如蒽基。
术语“5-14元杂芳基”应理解为包括这样的一价单环、双环或三环芳族环系：其具有5、6、7、8、9、10、11、12、13或14个环原子，特别是5或6或9或10个碳原子，且其包含1-5 个，优选1-3个独立选自N、O和S的杂原子，并且，另外在每一种情况下可为苯并稠合的。特别地，杂芳基选自噻吩基、呋喃基、吡咯基、噁唑基、噻唑基、咪唑基、吡唑基、异噁唑基、异噻唑基、噁二唑基、三唑基、噻二唑基、噻-4H-吡唑基等以及它们的苯并衍生物，例如苯并呋喃基、苯并噻吩基、苯并噁唑基、苯并异噁唑基、苯并咪唑基、苯并三唑基、吲唑基、吲哚基、异吲哚基等；或吡啶基、哒嗪基、嘧啶基、吡嗪基、三嗪基等，以及它们的苯并衍生物，例如喹啉基、喹唑啉基、异喹啉基等；或吖辛因基、吲嗪基、嘌呤基等以及它们的苯并衍生物；或噌啉基、酞嗪基、喹唑啉基、喹喔啉基、萘啶基、蝶啶基、咔唑基、吖啶基、吩嗪基、吩噻嗪基、吩噁嗪基等。
除非另有说明，杂环基或杂芳基包括其所有可能的异构形式，例如其位置异构体。因此，对于一些说明性的非限制性实例，吡啶基或亚吡啶基包括吡啶-2-基、亚吡啶-2-基、吡啶-3-基、亚吡啶-3-基、吡啶-4-基和亚吡啶-4-基；噻吩基或亚噻吩基包括噻吩-2-基、亚噻吩-2-基、噻吩-3-基和亚噻吩-3-基。
根据其分子结构，本发明的化合物可以是手性的，因此可能存在各种对映异构体形式。因而这些化合物可以以消旋体形式或光学活性形式存在。本发明的化合物或其中间体可以通过本领域技术人员公知的化学或物理方法分离为对映异构体化合物，或者以此形式用于合成。在外消旋的胺的情况中，通过与光学活性的拆分试剂反应，从混合物制得非对映异构体。适当的拆分试剂的示例是光学活性的酸，例如R和S形式的酒石酸、二乙酰酒石酸、二苯甲酰酒石酸、扁桃酸、苹果酸、乳酸、适当的N-保护的氨基酸(例如N-苯甲酰脯氨酸或N-苯磺酰基脯氨酸)或各种光学活性的樟脑磺酸。借助光学活性的拆分试剂(例如固定在硅胶上的二硝基苯甲酰基苯基甘氨酸、三乙酸纤维素或其它碳水化合物的衍生物或手性衍生化的异丁烯酸酯聚合物)，也可有利地进行色谱对映体拆分。用于此目的的适当的洗脱剂是含水或含醇的溶剂混合物，例如，己烷/异丙醇/乙腈。
本领域技术人员将理解，由于氮需要具有可用的孤对电子用于被氧化为氮氧化物，因此并非所有的含氮杂环都可以形成N-氧化物；本领域技术人员将识别能够形成N-氧化物的含氮杂环。本领域技术人员还将认识到叔胺能够形成N-氧化物。制备杂环和叔胺的N-氧化物的合成方法对于本领域技术人员而言是熟知的，所述合成方法包括用过氧酸如过氧乙酸和间氯过氧苯甲酸(MCPBA)、过氧化氢、烷基氢过氧化物如叔丁基氢过氧化物、过硼酸钠和双环氧乙烷(dioxirane)如二甲基双环氧乙烷氧化杂环和叔胺。这些制备N-氧化物的方法已在文献中广泛地描述和综述。
药学上可接受的盐可以是例如在链或环中具有氮原子的具有足够碱性的本发明的化合物的酸加成盐，例如与如下无机酸形成的酸加成盐：例如盐酸、氢氟酸、氢溴酸、氢碘酸、硫酸、焦硫酸、磷酸或硝酸，或硫酸氢盐、或者与如下有机酸形成的酸加成盐：例如甲酸、乙酸、乙酰乙酸、丙酮酸、三氟乙酸、丙酸、丁酸、己酸、庚酸、十一烷酸、月桂酸、苯甲酸、水杨酸、2-(4-羟基苯甲酰基)苯甲酸、樟脑酸、肉桂酸、环戊烷丙酸、二葡糖酸、3-羟基-2-萘甲酸、烟酸、扑酸、果胶酯酸、过硫酸、3-苯基丙酸、苦味酸、特戊酸、2-羟基乙磺酸、衣康酸、氨基磺酸、三氟甲磺酸、十二烷基硫酸、乙磺酸、苯磺酸、对甲苯磺酸、甲磺酸、2-萘磺酸、萘二磺酸、樟脑磺酸、柠檬酸、酒石酸、硬脂酸、乳酸、草酸、丙二酸、琥珀酸、苹果酸、己二酸、藻酸、马来酸、富马酸、D-葡糖酸、扁桃酸、抗坏血酸、葡庚酸、甘油磷酸、天冬氨酸、磺基水杨酸、半硫酸或硫氰酸。
另外，具有足够酸性的本发明的化合物的另一种适合的药学上可接受的盐是碱金属盐(例如钠盐或钾盐)、碱土金属盐(例如钙盐或镁盐)、铵盐，或与提供生理学可接受的阳离子的有机碱形成的盐，例如与如下物质形成的盐：钠离子、钾离子、N-甲基葡糖胺、二甲基葡糖胺、乙基葡糖胺、赖氨酸、二环己基胺、1,6-己二胺、乙醇胺、葡糖胺、葡甲胺、肌氨酸、丝氨醇、三羟基甲基氨基甲烷、氨基丙二醇、1-氨基-2,3,4-丁三醇。作为实例，所述药学上可接受的盐包括基团-COOH与如下物质形成的盐：钠离子、钾离子、钙离子、镁离子、N-甲基葡糖胺、二甲基葡糖胺、乙基葡糖胺、赖氨酸、二环己基胺、1,6-己二胺、乙醇胺、葡糖胺、葡甲胺、肌氨酸、丝氨醇、三羟基甲基氨基甲烷、氨基丙二醇、1-氨基-2,3,4-丁三醇。
另外，碱性含氮基团可用如下试剂季铵化：低级烷基卤化物，例如甲基、乙基、丙基和丁基氯化物、溴化物和碘化物；硫酸二烷基酯，例如硫酸二甲酯、硫酸二乙酯、硫酸二丁酯和硫酸二戊酯；长链卤化物，例如癸基、月桂基、肉豆蔻基和硬脂基氯化物、溴化物和碘化物；芳烷基卤化物如苄基和苯乙基溴化物等。作为实例，药学上可接受的盐包括盐酸盐、硫酸盐、硝酸盐、硫酸氢盐、氢溴酸盐、醋酸盐、草酸盐、柠檬酸盐、甲磺酸盐、甲酸盐或葡甲胺盐等。
由于本发明的化合物可存在多个成盐位点，所述药学上可接受的盐不仅包括本发明化合物其中1个成盐位点上形成的盐，而且还包括其中2、3或全部成盐位点上形成的盐。为此，所述药学上可接受的盐中式(I)化合物与成盐所需的酸的根离子(阴离子)或碱的阳离子摩尔比可以在较大的范围内变化，例如可以是4:1～1:4，如3:1、2:1、1:1、1:2、1:3等。
根据不同取代基的位置和性质，本发明的化合物还可以包含一个或多个不对称中心。不对称碳原子可以(R)或(S)构型存在，仅有一个不对称中心时，产生外消旋混合物，含有多个不对称中心时，得到非对映异构体混合物。在某些情况下，由于围绕特定键的旋转受阻还可能存在不对称性，例如该中心键连接特定化合物的两个被取代的芳族环。并且，取代基还可以顺式或反式异构的形式存在。
本发明化合物还包括其各自所有可能的立体异构体，其是单一立体异构体或所述立体异构体(例如R-异构体或S-异构体，或者E-异构体或Z-异构体)的任意比例的任意混合物的形式。可通过任意适合的现有技术方法(例如色谱法，特别是例如手性色谱法)实现本发明的化合物的单一立体异构体(例如单一对映异构体或单一非对映异构体)的分离。
术语“互变异构体”是指因分子中某一原子在两个位置迅速移动而产生的官能团异构体。本发明化合物可表现出互变异构现象。互变异构的化合物可以存在两种或多种可相互转化的种类。质子移变互变异构体来自两个原子之间共价键合的氢原子的迁移。互变异构体一般以平衡形式存在，尝试分离单一互变异构体时通常产生一种混合物，其理化性质与化合物的混合物是一致的。平衡的位置取决于分子内的化学特性。例如，在很多脂族醛和酮如乙醛中，酮型占优势；而在酚中，烯醇型占优势。本发明包含化合物的所有互变异构形式。
在本发明中，所涉及的化合物亦包括经同位素标记的化合物，所述经同位素标记的化合物与式I中所示的那些相同，但是其中一或多个原子被原子质量或质量数不同于通常天然存在的原子质量或质量数的原子替代。可掺入本发明的化合物的同位素的实例包括H、C、N、O、S、F及Cl的同位素，分别诸如 2H、 3H、 13C、 11C、 14C、 15N、 18O、 17O、 32P、 35S、 18F及 36Cl。含有上述同位素和/或其他原子的其他同位素的本发明的化合物、其前药、或者所述化合物或所述前药的药学上可接受的盐在本发明的范围内。本发明的某些经同位素标记的化 合物，例如掺入放射性同位素(诸如 3H和 14C)的化合物可用于药物和/或底物组织分布测定。氚(即 3H)和碳14(即 14C)同位素因易于制备和可检测性而成为特别优选的。再者，以较重的同位素(诸如氘，即 2H)替代可提供源自更高的代谢稳定性的某些治疗优势(例如增加的体内半衰期或减少的剂量需求)，并因此可在某些情况下是优选的。如权利要求所请求保护的本发明化合物可特别地限定以氘或氚替代。此外，取代基中出现的氢未单独列明术语氘或氚并不表示排除氘或氚，而是同样也可以包含氘或氚。
术语“有效量”或者“治疗有效量”是指足以实现预期应用(包括但不限于如下定义的疾病治疗)的本发明所述化合物的量。治疗有效量可以取决于以下因素而改变：预期应用(体外或者体内)，或者所治疗的受试者和疾病病症如受试者的重量和年龄、疾病病症的严重性和给药方式等，其可以由本领域普通技术人员容易地确定。具体剂量将取决于以下因素而改变：所选择的特定化合物、所依据的给药方案、是否与其它化合物组合给药、给药的时间安排、所给药的组织和所承载的物理递送系统。
术语“溶剂化物”是本发明的化合物的那些形式，其以固体或液体的状态通过与溶剂分子的配位作用形成配合物。水合物是溶剂化物的特定形式，其中配位作用是与水进行。在本发明中，优选的溶剂化物是水合物。进一步的，本发明通式I化合物的药学上可接受的溶剂化物(水合物)是指化合物I与化学计量学的一个或多个分子的水或其他溶剂形成的共晶和包合物。可用于溶剂化物的溶剂包括但不限于：水、甲醇、乙醇、乙二醇和醋酸。
术语“前药”或称为“药物前体”，代表化合物在体内转化为前述通式或具体化合物所示的化合物。这样的转化受前体药物在血液中水解或在血液或组织中经酶转化为母体结构的影响。本发明前药可以是酯，在本发明中酯可以作为前药的有苯酯类，脂肪族酯类，酰氧基甲基酯类，碳酸酯，氨基甲酸酯类和氨基酸酯类。例如本发明里的一个化合物包含羟基/羧基，即可以将其酰化得到前体药物形式的化合物。其他的前药形式包括磷酸酯，如这些磷酸酯类化合物是经母体上的羟基磷酸化得到的。
有益效果
1)本发明提供了新颖的通式I化合物，具有对整合素的良好抑制作用；
2)本发明化合物对多种整合素亚型均具有抑制作用，其中一些化合物对αVβ1，αVβ3，αVβ5，αVβ6，和αVβ8，α5β1，α8β1中的一种或多种具有明显的抑制作用。
具体实施方式
下文将结合具体实施例对本发明的技术方案做更进一步的详细说明。应当理解，下列实施例仅为示例性地说明和解释本发明，而不应被解释为对本发明保护范围的限制。凡基于本发明上述内容所实现的技术均涵盖在本发明旨在保护的范围内。
除非另有说明，以下实施例中使用的原料和试剂均为市售商品，或者可以通过已知方法制备。
仪器仪表和一般方法
采用氘代试剂(DMSO-d 6、CDCl 3、CD 3OD)在Bruker Ascend 400光谱仪上记录 1H和 19F NMR。氘代溶剂或TMS作为内标。化学位移以ppm表示，耦合常数(J)以Hz表示，裂分方式有s(单峰)，d(双峰)，t(三重峰)，q(四重峰)，m(多重峰)，br(宽峰)。LC-MS采用安 捷伦1260-6120系统配备Waters Cortecs C18，2.7μm,4.6×30mm柱子；HPLC采用Waters Acquity UPLC H-class仪器配备Acquity BEH C18,1.7μm,50×2.1mm柱子。采用制备型HPLC(Kromasil-C18(100×21.2mm，5μm)柱子、Xtimate-C18(250×30mm)柱子或者Xbridge-C18(150×19mm，5μm)柱子)、手性制备型SFC(Daicel ChiralPak IG(250mm×30mm,10μm)柱子、(S,S)Whelk-O1(100×4.6mm)柱子、Daicel Chiralpak IC-3(50×4.6mm)柱子、Daicel Chiralpak AD-H(250×4.6mm)柱子和Chirex S-VAL R-NEA柱子(250×4.6mm))纯化或者拆分最终化合物。
通用中间体的合成步骤示例：
中间体Int-A的合成：
Figure PCTCN2021078537-appb-000067
步骤1：7-甲基-1，2，3，4-四氢-1，8-萘啶
Figure PCTCN2021078537-appb-000068
在N 2气氛下，向2-甲基-1，8-萘啶(48.0g,333mmol)的乙醇(840mL)溶液中加入10％的钯碳(10g)。反应瓶抽真空，然后在1个大气压的氢气气氛下于25℃搅拌16小时。然后将钯碳通过硅藻土过滤并用乙醇(50mL)洗涤。滤液在真空下浓缩得到标题化合物(48.3g,97％)，为淡黄色固体。 1H NMR(400MHz,DMSO-d 6)δ7.00(d,J＝7.2Hz,1H),6.25(d,J＝7.1Hz,1H),6.24(br,1H),3.25–3.21(m,2H),2.59(t,J＝6.2Hz,2H),2.17(s,3H),1.77–1.71(m,2H).LC-MS:ESI m/z 149.11[M+H] +；C 9H 12N 2计算值148.10.
步骤2：7-甲基-3,4-二氢-1,8-萘啶-1(2H)-羧酸叔丁酯
Figure PCTCN2021078537-appb-000069
在N 2气氛下，7-甲基-1，2，3，4-四氢-1，8-萘啶(48.3g,324mmol)和Boc酸酐(200mL)的混合物在50℃搅拌16小时。溶剂在真空下除去，残留物用柱层析(0～100％乙酸乙酯的石油醚溶液)纯化得到题目化合物(42.6g,60％)，为淡黄色固体。 1H NMR(400MHz,DMSO-d 6)δ7.39(d,J＝7.6Hz,1H),6.89(d,J＝7.6Hz,1H),3.65–3.59(m,2H),2.68(t,J＝6.6Hz,2H),2.36(s,3H),1.84–1.78(m,2H),1.44(s,9H).LC-MS:ESI m/z 249.2[M+H] +；C 14H 20N 2O 2计算值248.15.
步骤3：7-(2-甲氧基-2-氧代乙基)-3,4-二氢-1,8-萘啶-1(2H)-羧酸叔丁酯
Figure PCTCN2021078537-appb-000070
在-78℃，N 2气氛下，向7-甲基-3,4-二氢-1,8-萘啶-1(2H)-羧酸叔丁酯(15.0g,60.4mmol)和碳酸二甲酯(19.0g,211mmol)的四氢呋喃(150mL)溶液中滴加LDA的四氢呋喃溶液(1.0N,90mL,90mmol)。得到的溶液在-78℃搅拌40分钟后，反应用饱和氯化铵水溶液(200mL)淬灭。水相用乙酸乙酯(150mL×3)萃取。合并的有机相用饱和食盐水洗涤，用硫酸钠干燥，过滤。滤液在真空下浓缩。残留物用柱层析(0～100％乙酸乙酯的石油醚溶液)纯化得到标题化合物(15.2g,82％)，为红色油状物。 1H NMR(400MHz,DMSO-d 6)δ7.47(d,J＝7.6Hz,1H),6.99(d,J＝7.6Hz,1H),3.71(s,2H),3.63–3.61(m,2H),3.60(s,3H),2.71(t,J＝6.6Hz,2H),1.87–1.75(m,2H),1.41(s,9H).LC-MS:ESI m/z 307.2[M+H] +；C 16H 22N 2O 4计算值306.16.
步骤4：7-(2-羟乙基)-3,4-二氢-1,8-萘啶-1(2H)-羧酸叔丁酯
Figure PCTCN2021078537-appb-000071
在N 2气氛下，向7-(2-甲氧基-2-氧代乙基)-3,4-二氢-1,8-萘啶-1(2H)-羧酸叔丁酯((15.2g,49.6mmol)的四氢呋喃(120mL)溶液中加入硼氢化锂的四氢呋喃溶液(2N,32mL,64mmol)。反应混合物在40℃搅拌2小时并用冰浴冷却后，反应用水淬灭(50mL)。水溶液用乙酸乙酯(100mL×3)萃取。合并的有机相用饱和食盐水洗涤，用硫酸钠干燥，过滤。滤液在减压下浓缩得到标题化合物(13.3g,86％)，为黄色油状物。 1H NMR(400MHz,DMSO-d 6)δ7.41(d,J＝7.6Hz,1H),6.90(d,J＝7.6Hz,1H),4.74(t,J＝5.5Hz,1H),3.74–3.69(m,2H),3.65–3.61(m,2H),2.78(t,J＝6.7Hz,2H),2.68(t,J＝6.6Hz,2H),1.83–1.77(m,2H),1.44(s,9H).LC-MS:ESI m/z 279.2[M+H] +；C 15H 22N 2O 3计算值278.16.
步骤5：7-(2-(甲苯磺酰氧基)乙基)-3,4-二氢-1,8-萘啶-1(2H)-羧酸叔丁酯
Figure PCTCN2021078537-appb-000072
在N 2气氛下，向7-(2-羟乙基)-3,4-二氢-1,8-萘啶-1(2H)-羧酸叔丁酯(13.3g,47.8mmol)和三乙胺(13.2mL)的1，2-二氯乙烷(130mL)溶液中加入对甲苯磺酰氯(10.9g,57.3mmol)和4-二甲氨基吡啶(291mg,2.39mmol)。得到的溶液在25℃搅拌16小时后，反应用水(150mL)淬灭。水相用二氯甲烷(50mL×3)萃取。合并的有机相用饱和食盐水洗涤，用硫酸钠干燥，过滤。滤液在真空下浓缩。残留物用柱层析(0～50％乙酸乙酯的石油醚溶液)纯化得到标题化合物(11.2g,54％)，为黄色油状物。LC-MS:ESI m/z 433.3[M+H] +；C 22H 28N 2O 5S计算值432.17.
中间体Int-B的合成：
Figure PCTCN2021078537-appb-000073
7-(2-碘乙基)-3,4-二氢-1,8-萘啶-1(2H)-羧酸叔丁酯
在0℃，N 2气氛下，向三苯基膦(3.8g，14mmol)，碘(3.6g，14mmol)，1H-咪唑(0.97g，14mmol)的四氢呋喃(10mL)溶液中滴加7-(2-羟乙基)-3,4-二氢-1,8-萘啶-1(2H)- 羧酸叔丁酯(4.0g，14mmol)的四氢呋喃溶液(20mL)。然后将反应在0℃下搅拌1小时。将该混合物倒入水(50mL)中，并用乙酸乙酯(30mL×2)萃取。合并的有机相用盐水洗涤，经硫酸钠干燥，过滤并浓缩至干。残留物通过硅胶色谱柱(石油醚：乙酸乙酯＝10：1-1：1)纯化粗产物，得到标题化合物(4.2g，76％)，为白色固体。 1H NMR(400MHz,CDCl 3)δ7.34(d,J＝7.6Hz,1H),6.83(d,J＝7.6Hz,1H),3.82–3.72(m,2H),3.53(t,J＝7.2Hz,2H),3.28(t,J＝7.2Hz,2H),2.74(t,J＝6.4Hz,2H),1.97–1.88(m,2H),1.52(s,9H).
中间体Int-C的合成：
Figure PCTCN2021078537-appb-000074
步骤1：5-乙酰戊酸甲酯
Figure PCTCN2021078537-appb-000075
在N 2气氛下，向5-乙酰戊酸(15.0g,104mmol)的甲醇(150mL)溶液中加入浓硫酸(98％,5mL)，得到的反应混合物在80℃搅拌14小时。在浓缩干燥后，残留物溶解在乙酸乙酯(200mL)中，有机溶液用水和饱和食盐水洗涤，用硫酸钠干燥，过滤。滤液在真空下浓缩得到标题化合物(16.5g,100％)，为黄色油状物，无需进一步纯化直接用于下一步反应。
1H NMR(400MHz,CDCl 3)δ3.67(s,3H),2.45(t,J＝6.8Hz,2H),2.33(t,J＝7.1Hz,2H),2.14(s,3H),1.65–1.58(m,4H).LC-MS:ESI m/z 159.1[M+H] +；C 8H 14O 3计算值158.09.
步骤2：5-(1,8-萘啶-2-基)戊酸甲酯
Figure PCTCN2021078537-appb-000076
在N 2气氛下，将5-乙酰戊酸甲酯(16.5g,104mmol),2-氨基-3-吡啶甲醛(13.5g,111mmol)和L-脯氨酸(6.0g,52mmol)加入到乙醇(150mL)中，得到的反应混合物在80℃搅拌16小时，然后浓缩干燥，残留物用柱层析(0-6％甲醇的二氯甲烷溶液)纯化得到标题化合物(11.2g,54％)，为黄色固体。 1H NMR(400MHz,CDCl 3)δ9.01(m,1H),8.09(dd,J＝8.1,2.0Hz,1H),8.03(d,J＝8.3Hz,1H),7.37(dd,J＝8.1,4.3Hz,1H),7.32(d,J＝8.3Hz,1H),3.59(s,3H),3.03–2.97(m,2H),2.32(t,J＝7.5Hz,2H),1.89(m,2H),1.73–1.65(m,2H).LC-MS:ESI m/z 245.2[M+H] +；C 14H 16N 2O 2计算值244.12.
步骤3：5-(5,6,7,8-四氢-1,8-萘啶-2-基)戊酸甲酯
Figure PCTCN2021078537-appb-000077
向5-(1,8-萘啶-2-基)戊酸甲酯(12.8g,52.4mmol)的甲醇(150mL)溶液中加入10％的钯碳(3.0g)，得到的混合物在1个大气压的氢气气氛下于25℃搅拌20小时。在通过硅藻土过滤并用甲醇(500mL)洗脱后，滤液在真空下浓缩得到粗品标题化合物(13.0g,100％)，为黄色油状物，得到的粗品直接用于下一步反应。LC-MS:ESI m/z 249.27[M+H] +；C 14H 20N 2O 2计算值248.15.
步骤4：7-(5-甲氧基-5-氧戊基)-3,4-二氢-1,8-萘啶-1(2H)-羧酸叔丁酯
Figure PCTCN2021078537-appb-000078
在N 2气氛下，5-(5,6,7,8-四氢-1,8-萘啶-2-基)戊酸甲酯(13.0g,52.3mmol)在Boc酸酐(60mL,262mmol)的混合物在50℃搅拌16小时。在冷却到室温后，残留物用柱层析(0-60％乙酸乙酯的石油醚溶液)纯化得到标题化合物(17.0g,93％)，为黄色油状物。
1H NMR(400MHz,CDCl 3)δ7.28(d,J＝7.6Hz,1H),6.80(d,J＝7.6Hz,1H),3.75(t,J＝6.6,2H),3.66(s,3H),2.76–2.67(m,4H),2.35(t,J＝7.3Hz,2H),1.95–1.87(m,2H),1.80–1.70(m,4H),1.51(s,9H).LC-MS:ESI m/z 349.3[M+H] +；C 19H 28N 2O 4计算值348.20.
步骤5：5-(8-(叔丁氧羰基)-5,6,7,8-四氢-1,8-萘啶-2-基)戊酸
Figure PCTCN2021078537-appb-000079
向7-(5-甲氧基-5-氧戊基)-3,4-二氢-1,8-萘啶-1(2H)-羧酸叔丁酯(6.7g,19mmol)在甲醇(60mL)和水(10mL)溶液中加入氢氧化锂(553mg,23.1mmol)。得到的混合物在25℃搅拌16小时，然后在真空下浓缩得到锂盐形式的粗品标题化合物(6.5g,100％)，为黄色固体，无需进一步纯化直接用于下一步反应。LC-MS:ESI m/z 335.3[M+H] +；C 18H 26N 2O 4计算值334.19.
中间体Int-D的合成：
Figure PCTCN2021078537-appb-000080
步骤1：7-(5-(甲氧基(甲基)氨基)-5-氧戊基)-3,4-二氢-1,8-萘啶-1(2H)-羧酸叔丁酯
Figure PCTCN2021078537-appb-000081
向5-(8-(叔丁氧基羰基)-5,6,7,8-四氢-1,8-萘啶-2-基)戊酸(6.5g,19mmol),N，O-二甲基羟胺盐酸盐(2.25g,23.1mmol)和三乙胺(11mL,77mmol)在二氯甲烷(60mL)的混合物中加入HBTU(11g,29mmol)，得到的混合物在N 2氛围下于25℃搅拌16小时。反应用饱和碳酸氢钠水溶液(50mL)淬灭后，水相用二氯甲烷(50mL×2)萃取。合并的有机相用饱和食盐水洗涤，用硫酸钠干燥，过滤。滤液在真空下浓缩。残留物用柱层析(0～80％乙酸乙酯的石油醚溶液)纯化得到标题化合物(6.0g,83％)，为淡黄色油状物。 1H NMR(400MHz,DMSO-d 6)δ7.40(d,J＝7.6Hz,1H),6.88(d,J＝7.6Hz,1H),3.66–3.57(m,2H),3.63(s,3H),3.06(s,3H),2.68–2.64(m,2H),2.61–2.57(m,2H),2.38(t,J＝7.3Hz,2H),1.85–1.76(m,2H),1.71–1.63(m,2H),1.57–1.49(m,2H),1.43(s,9H).LC-MS:ESI m/z 378.4[M+H] +；C 20H 31N 3O 4计算值377.23.
步骤2：7-(5-氧杂戊基)-3,4-二氢-1,8-萘啶-1(2H)-羧酸叔丁酯
Figure PCTCN2021078537-appb-000082
在-78℃，N 2气氛下，向7-(5-(甲氧基(甲基)氨基)-5-氧戊基)-3,4-二氢-1,8-萘啶-1(2H)-羧酸叔丁酯(1.0g,2.7mmol)的四氢呋喃(20mL)溶液中滴加入DIBAL-H的四氢呋喃溶液(4.0mL,4N,4.0mmol)，得到的混合物在-78℃搅拌3小时。反应用冰水(40mL)淬灭后，用乙酸乙酯(50mL×2)萃取。合并的有机相用饱和食盐水洗涤，用硫酸钠干燥，过滤。滤液在真空下浓缩得到标题化合物(744mg,87％)，为淡黄色油状物，得到的粗品直接用于下一步合成。 1H NMR(400MHz,CDCl 3)δ9.77(t,J＝1.8Hz,1H),7.29(d,J＝7.6Hz,1H),6.80(d,J＝7.6Hz,1H),3.75(t,J＝6.6,2H),2.76–2.70(m,4H),2.47(td,J＝7.2,1.8Hz,2H),1.96–1.88(m,2H),1.84–1.67(m,4H),1.51(s,9H).LC-MS:ESI m/z 319.26[M+H] +；C 18H 26N 2O 3计算值318.19.
中间体Int-E的合成：
Figure PCTCN2021078537-appb-000083
步骤1：(S)-3-(((苄氧基)羰基)氨基)吡咯烷-1-羧酸叔丁酯
Figure PCTCN2021078537-appb-000084
在0℃，N 2气氛下，向(S)-3-氨基吡咯烷-1-羧酸叔丁酯(10.0g,53.7mmol)和N,N'-二异丙基乙胺(17.8mL,107mmol)的二氯甲烷(150mL)溶液中滴加入CbzCl(11.0g,64.4mmol)，得到的混合物于25℃搅拌16小时。反应用饱和碳酸氢钠水溶液(100mL)淬灭后，得到的混合物用二氯甲烷(50mL×2)萃取。有机相用饱和食盐水洗涤，用硫酸钠干燥，过滤。滤液在真空下浓缩。残留物用柱层析(0～60％乙酸乙酯的石油醚溶液)纯化得到标题化合物(13.1g,77％)，为白色油状物。LC-MS:ESI m/z 321.3[M+H] +；C 17H 24N 2O 4计算值320.17.
步骤2：(S)-3-(((苄氧基)羰基)(甲基)氨基)吡咯烷-1-羧酸叔丁酯的合成
Figure PCTCN2021078537-appb-000085
在0℃，N 2气氛下，向(S)-3-(((苄氧基)羰基)氨基)吡咯烷-1-羧酸叔丁酯(13.1g,40.9mmol)的四氢呋喃(150mL)溶液中加入60％分布在矿物油中的氢化钠(2.45g,61.3mmol)，得到的混合物在0℃搅拌15分钟。此后，碘甲烷(5.1mL,82mmol)加入到溶液中，得到的混合物在25℃搅拌16小时。反应用冰的饱和氯化铵水溶液淬灭后，混合物用乙酸乙酯(50mL×2)萃取。有机相用饱和食盐水洗涤，用硫酸钠干燥，过滤。滤液在真空下浓缩。残留物用柱层析(0-40％乙酸乙酯的石油醚溶液)纯化得到标题化合物(12.8g,94％)，为白色油状物。LC-MS:ESI m/z 335.3[M+H] +；C 18H 26N 2O 4计算值334.19.
步骤3：(S)-甲基(吡咯烷基-3-基)氨基甲酸酯
Figure PCTCN2021078537-appb-000086
向(S)-3-(((苄氧羰基)甲基)氨基)吡咯烷-1-羧酸叔丁酯(5.7g,17mmol)的二氯甲烷(100mL)溶液中加入三氟乙酸(25mL)，得到的混合物在25℃搅拌16小时。溶剂在真空下移除，得到粗品标题化合物(4.0g,100％)，为黄色油状物。
步骤4：(S)-7-(5-(3-(((苄氧基)羰基)(甲基)氨基)吡咯烷-1-基)戊基)-3,4-二氢-1,8-萘啶-1(2H)-羧酸叔丁酯
Figure PCTCN2021078537-appb-000087
在N 2气氛下，向(S)-甲基(吡咯烷基-3-基)氨基甲酸酯(4.0g,17mmol)和7-(5-氧杂戊基)-3,4-二氢-1,8-萘啶-1(2H)-羧酸叔丁酯(8.15g,25.6mmol)的甲醇(100mL)溶液中加入三乙酰氧基硼氢化钠(10.9g,52.2mmol)，得到的混合物于25℃搅拌18小时。此后，氰基硼氢化钠(1.6g,26mmol)加入到溶液中，得到的混合物在25℃搅拌14小时。浓缩至干燥后，将残留物溶解在乙酸乙酯(80mL)中，饱和碳酸氢钠水溶液洗涤(80mL)。有机相用饱和食盐水洗涤，用硫酸钠干燥，过滤。滤液在真空下浓缩。残留物用柱层析(0-8％甲醇的二氯甲烷溶液)纯化得到标题化合物(12.8g,94％)，为黄色油状物。LC-MS:ESI m/z 537.3[M+H] +；C 31H 44N 4O 4.计算值536.34.
步骤5：(S)-7-(5-(3-(甲基氨基)吡咯烷-1-基)戊基)-3,4-二氢-1,8-萘啶-1(2H)-羧酸叔丁酯
Figure PCTCN2021078537-appb-000088
在N 2气氛下，向(S)-7-(5-(3-(((苄氧羰基)甲基)氨基)吡咯烷-1-基)戊基)-3,4-二氢-1,8-萘啶-1(2H)-羧酸叔丁酯(5.5g,10mmol)的甲醇(80mL)溶液中加入10％钯碳(1.0g)。N 2被氢气置换后，得到的混合物在25℃搅拌16小时。混合物通过硅藻土过滤后，滤液在真空下浓缩得到粗品标题化合物(4.1g,99％)，为黄色油状物，粗品未经纯化直接用于下一步反应。LC-MS:ESI m/z 403.2[M+H] +,C 23H 38N 4O 2计算值402.30.
按照与中间体Int-C相似的合成路线，合成中间体Int-F：
Figure PCTCN2021078537-appb-000089
向7-(3-乙氧基-3-氧丙基)-3,4-二氢-1,8-萘啶-1(2H)-羧酸叔丁酯(4.1g，12.2mmol)的甲醇(40mL)和水(10mL)溶液中加入LiOH(350mg，14.6mmol)，并将得到的混合物在25℃下搅拌16小时。真空浓缩得到浅黄色固体的锂盐形式的标题化合物(3.7g，100％)，其无需进一步纯化即直接用于下一步。LC-MS:ESI m/z 307.2[M+H] +,C 16H 22N 2O 4.计算值306.16.
按照与中间体Int-D相似的合成路线，合成中间体Int-G：
Figure PCTCN2021078537-appb-000090
在-78℃，N 2气氛下，将DIBAL-H(1M,17.1mL,17.1mmol)逐滴加入到7-(3-(甲氧基(甲基)氨基)-3-氧丙基)-3,4-二氢-1,8-萘啶-1(2H)-羧酸叔丁酯(2.0g，5.7mmol)的四氢呋喃(20mL)溶液中，得到的混合物在搅拌3小时。用冰水(30mL)淬灭后，将反应物用乙酸乙酯(20mL)萃取两次，并将合并的有机相用盐水洗涤，用硫酸钠干燥，过滤并浓缩，得到浅黄色的标题化合物的粗品(1.7g，100％)。 1H NMR(400MHz,CDCl 3)δ9.88(t,J＝1.2Hz,1H),7.29(d,J＝7.6Hz,1H),6.84(d,J＝7.6Hz,1H),3.76–3.73(m,2H),3.09–3.0(m,2H),2.94–2.91(m,2H),2.72(t,J＝6.6Hz,2H),1.95–1.88(m,2H),1.52(s,9H).LC-MS:ESI m/z 291.2[M+H] +；C 16H 22N 2O 3计算值290.16.
按照与中间体Int-E相似的合成路线，合成中间体Int-H：
Figure PCTCN2021078537-appb-000091
向(S)-7-(3-(3-(((苄氧基)羰基)(甲基)氨基)吡咯烷-1-基)-丙基)-3,4-二氢-1,8-萘啶-1(2H)-羧酸盐(2.4g，4.71mmol)的甲醇(50mL)溶液中加入10％的钯碳(250mg)。该混合物在H 2气氛下于40℃下反应16小时。将混合物过滤并用甲醇洗涤。真空浓缩滤液，得到黄色油状的标题化合物(1.4g，79％)。LC-MS:ESI m/z 375.2[M+H] +；C 21H 34N 4O 2计算值374.27.
中间体Int-I的合成：
Figure PCTCN2021078537-appb-000092
步骤1：3-(((R)-2-甲氧基-2-氧代-1-苯乙基)氨基)吡咯烷基-1-羧酸叔丁酯
Figure PCTCN2021078537-appb-000093
在N 2气氛下，向(R)-2-氨基-2-苯乙酸甲酯盐酸盐(5.0g，24.8mmol)和1-叔丁氧羰基-3-吡咯烷酮(5.5g，29.75mmol)的无水甲醇(60mL)溶液中，加入HOAc(5mL)，并将所得混合物于25℃搅拌16小时。然后再加入氰基硼氢化钠(2.34g，37.19mmol)，并将所得混 合物继续搅拌14小时。浓缩至干后，将残余物溶在水(50mL)中，并用乙酸乙酯(50mL)萃取。有机相用饱和碳酸氢钠(60mL)和盐水洗涤，经硫酸钠干燥，过滤并浓缩至干。残余物通过快速柱色谱法纯化(0-60％乙酸乙酯的石油醚溶液)，得到标题化合物(3.7g，45％)，为浅黄色油状物。LC-MS:ESI m/z 335.1[M+H] +,C 18H 26N 2O 4.计算值334.19.
步骤2：3-(((R)-2-甲氧基-2-氧-1-苯基乙基)(甲基)氨基)吡咯烷-1-羧酸叔丁酯
Figure PCTCN2021078537-appb-000094
在N 2气氛下，向3-((((R)-2-甲氧基-2-氧代-1-苯乙基)氨基)吡咯烷基-1-羧酸叔丁酯(2.2g，6.58mmol)和多聚甲醛(988mg，32.89mmol)的甲醇溶液(50mL)加入三乙酰氧基硼氢化钠(4.18g，19.74mmol)，将得到的混合物于25℃搅拌18小时。然后再加入氰基硼氢化钠(465mg，7.4mmol)，并将所得混合物继续搅拌14小时。浓缩至干后，将残余物溶解在乙酸乙酯(40mL)中，并用饱和碳酸氢钠(60mL)洗涤。有机相再用盐水洗涤，经硫酸钠干燥，过滤并浓缩至干。残余物通过快速柱色谱法纯化(0-60％乙酸乙酯的石油醚溶液)，得到标题化合物(1.58g，69％)，为浅黄色油状物。LC-MS:ESI m/z 349.2[M+H] +,C 19H 28N 2O 4.计算值348.20.
步骤3：(2R)-2-(甲基(吡咯烷-3-基)氨基)-2-苯乙酸甲酯
Figure PCTCN2021078537-appb-000095
向3-(((R)-2-甲氧基-2-氧代-1-苯基乙基)(甲基)氨基)吡咯烷-1-羧酸叔丁酯(1.58g，4.53mmol)的二氯甲烷(20mL)溶液中，加入盐酸/1，4-二氧六环(4N，4mL)溶液，并将所得混合物在25℃下搅拌16小时。真空除去溶剂，得到黄色固体形式的标题化合物(1.13g，100％)。LC-MS:ESI m/z 249.2[M+H] +,C 14H 20N 2O 2计算值248.15.
具体化合物的合成如下：
实施例1.化合物1的合成
Figure PCTCN2021078537-appb-000096
步骤1. 7-(3-(3-(((R)-2-甲氧基-2-氧-1-苯基乙基)氨基)吡咯烷-1-基)丙基)-3,4-二氢-1,8-萘啶-1(2H)-羧酸叔丁酯
Figure PCTCN2021078537-appb-000097
向(2R)-2-苯基-2-(吡咯烷-3-基氨基)乙酸甲酯盐酸盐(Int-I，140mg，0.6mmol)和7-(5-氧代戊基)-3,4-二氢-1,8-萘啶-1(2H)-羧酸叔丁酯(Int-G，203mg，0.7mmol)的甲醇(5mL)溶液中加入三乙酰氧基硼氢化钠(191mg，0.9mmol)，将得到的混合物在N 2气氛下于25℃搅拌18小时。然后再加入氰基硼氢化钠(57mg，0.9mmol)，并将得到的混合物在N 2气氛下在25℃继续搅拌14小时。浓缩至干后，将残余物用饱和NaHCO 3(15mL)稀释，并以乙酸乙酯(15mL×2)萃取。合并的有机相用盐水洗涤，经硫酸钠干燥，过滤并浓缩至干。残余物通过快速柱色谱法纯化(0-8％甲醇的二氯甲烷溶液)，得到标题化合物(120mg，39％)，为黄色油状物。LC-MS:ESI m/z 509.4[M+H] +,C 29H 40N 4O 4.计算值508.30.
步骤2.(2R)-2-苯基-2-((1-(3-(5,6,7,8-四氢-1,8-萘啶-2-基)丙基)吡咯烷-3-基)氨基)乙酸甲酯
Figure PCTCN2021078537-appb-000098
向7-(3-(3-(((R)-2-甲氧基-2-氧-1-苯基乙基)氨基)吡咯烷-1-基)丙基)-3,4-二氢-1,8-萘啶-1(2H)-羧酸叔丁酯(60mg，0.12mmol)的二氯甲烷(1mL)溶液中加入TFA(1mL)，并将所得混合物在25℃下搅拌16小时。真空除去溶剂，得到标题化合物(24mg，50％)，为浅黄色油状的TFA盐形式，其无需进一步纯化即可用于下一步。LC-MS:ESI m/z 409.2[M+H] +,C 24H 32N 4O 2.计算值408.25.
步骤3.(2R)-2-苯基-2-((1-(3-(5,6,7,8-四氢-1,8-萘啶-2-基)丙基)吡咯烷-3-基)氨基)乙酸
Figure PCTCN2021078537-appb-000099
将TFA盐形式的(2R)-2-苯基-2-((1-(3-(5,6,7,8-四氢-1,8-萘啶-2-基)丙基)吡咯烷-3-基)氨基)乙酸甲酯(24mg，0.06mmol)溶于甲醇(2mL)和水(0.5mL)中，加入NaOH(5mg，0.12mmol)，并将所得混合物在25℃下搅拌16小时。用1N盐酸(0.4mL)中和后，将混合物浓缩并将残余物通过制备HPLC在以下条件下纯化[柱：Kromasil 100-5-C18，30×150mm；流动相：1-100％乙腈的水溶液(含0.1％甲酸)，14分钟]，得到标题化合物(4.3mg，18％)，为白色固体。 1H NMR(400MHz,CD 3OD)δ7.56–7.19(m,6H),6.53(d,J＝7.3Hz,0.5H),6.42(d,J＝7.3Hz,0.5H),4.46(s,0.5H),4.41(s,0.5H),3.80–3.76(m,0.5H),3.61–3.57(m,0.5H),3.48–3.38(m,2H),3.18–3.09(m,1H),3.04–2.79(m,4H),2.77–2.64(m,3H),2.59–2.50(m,1H),2.33–2.25(m,1H),2.11–1.66(m,6H).LC-MS:ESI m/z 395.1[M+H] +；C 23H 30N 4O 2计算值394.24.
实施例2 参照化合物1的合成方法，可以得到以下化合物：
化合物2(甲酸盐)： 1H NMR(400MHz,CD 3OD)δ7.56–7.45(m,3H),7.44–7.31(m,3H),6.58(dd,J＝7.3,4.5Hz,1H),4.61(s,0.67H),4.52(s,0.33H),3.68–3.66(m,2H),3.53–3.45(m,2H),3.34–3.32(m,1H),3.27–3.10(m,4H),2.83–2.68(m,4H),2.40–2.20(m,2H),1.97–1.72(m,6H).LC-MS:ESI m/z 409.1[M+H] +；C 24H 32N 4O 2计算值408.25.
化合物3(甲酸盐)： 1H NMR(400MHz,CD 3OD)δ7.53–7.46(m,3H),7.42–7.30(m,3H),6.52(d,J＝7.2Hz,1H),4.45–4.42(m,1H),3.70–3.64(m,1H),3.50–3.32(m,4H),3.17–3.07(m,1H),3.00–2.61(m,7H),2.28–2.20(m,1H),2.08–1.98(m,1H),1.94–1.80(m,3H),1.73–1.53(m,4H),1.48–1.41(m,1H).LC-MS:ESI m/z 423.2[M+H] +；C 25H 34N 4O 2计算值422.27.
化合物4(甲酸盐)： 1H NMR(400MHz,CD 3OD)δ7.44–7.33(m,6H),6.52–6.47(m,1H),4.54–4.52(m,1H),3.41–3.32(m,4H),3.23–3.12(m,2H),2.92–2.72(m,6H),2.30–1.85(m,5H).LC-MS:ESI m/z 381.0[M+H] +；C 22H 28N 4O 2计算值380.22.
实施例3.化合物5的合成
Figure PCTCN2021078537-appb-000100
步骤1：(3S)-3-((2-甲氧基-2-氧代-1-苯基乙基)氨基)吡咯烷-1-羧酸叔丁酯
Figure PCTCN2021078537-appb-000101
在N 2气氛下，向(S)-3-氨基吡咯烷-1-羧酸叔丁酯(948mg，5.09mmol)和2-溴-2-苯基乙酸甲酯(1.4g，6.1mmol)的乙腈(30mL)溶液添加K 2CO 3(1.37g,12.7mmol),NaI(763mg，5.1mmol)。将所得混合物在60℃下搅拌16小时后，真空除去溶剂。残余物通过快速柱色谱法纯化(0-30％乙酸乙酯的石油醚溶液)，得到标题化合物(1.5g，74％)，为无色油状物。LC-MS:ESI m/z:335.4[M+H] +；C 18H 26N 2O 4计算值334.19.
步骤2：(3S)-3-((2-甲氧基-2-氧-1-苯基乙基)(甲基)氨基)吡咯烷-1-甲酸叔丁酯
Figure PCTCN2021078537-appb-000102
在N 2气氛下，向(3S)-3-((2-甲氧基-2-氧代-1-苯基乙基)氨基)吡咯烷-1-羧酸叔丁酯(1.5g，4.5mmol)和多聚甲醛(538mg，17.9mmol)的甲醇(30mL)溶液中加入三乙酰氧基硼氢化钠(1.9g，9.0mmol)。将混合物在25℃下搅拌16小时，然后再加入氰基硼氢化钠(422mg，6.72mmol)。将反应混合物在25℃下进一步搅拌4小时。真空除去溶剂。将残余物用饱和NaHCO 3水溶液(30mL)稀释，并用二氯甲烷(30mL×2)萃取。合并的有机相用盐水(30mL)洗涤，经硫酸钠干燥，并过滤。将滤液在减压下浓缩。残余物通过快速柱色谱法纯化(0-3％的甲醇的二氯甲烷溶液)，得到标题化合物(1.4g，89％)，为无色油状物。LC-MS:ESI m/z:349.3[M+H] +；C 19H 28N 2O 4计算值348.20.
步骤3：2-(甲基((S)-吡咯烷基-3-基)氨基)-2-苯基乙酸甲酯
Figure PCTCN2021078537-appb-000103
向(3S)-3-((2-甲氧基-2-氧代-1-苯基乙基)(甲基)氨基)吡咯烷-1-甲酸叔丁酯(1.4g，4.0mmol)的二氯甲烷(5mL)溶液中，加入盐酸/1，4-二氧六环(4N，10mL)溶液。所得溶液在25℃下搅拌3小时后，真空除去溶剂。将残余物溶解在甲醇和二氯甲烷的混合溶剂中[30mL，1∶1(v/v)]，然后加入固体NaHCO 3(1g)。将得到的悬浮液在25℃下搅拌1小时，然后过滤。减压浓缩滤液，得到标题化合物(950mg，95％)，为无色油状物。LC-MS:ESI m/z:249.3[M+H] +；C 14H 20N 2O 2计算值248.15.
步骤4：7-(3-((3S)-3-((2-甲氧基-2-氧-1-苯乙基)(甲基)氨基)吡咯烷-1-基)丙基)-3,4-二氢-1,8-萘啶-1(2H)-羧酸叔丁基酯
Figure PCTCN2021078537-appb-000104
在N 2气氛下，向2-(甲基((S)-吡咯烷-3-基)氨基)-2-苯基乙酸甲酯(500mg，2.0mmol)和7-(3-氧丙基)-3,4-二氢-1,8-萘啶-1(2H)-羧酸叔丁酯(639mg，2.2mmol)的甲醇(12mL)溶液中加入三乙酰氧基硼氢化钠(848mg，4.0mmol)。将混合物在25℃下搅拌16小时，然后再加入氰基硼氢化钠(188mg，3.0mmol)。将反应混合物在25℃下进一步搅拌4小时。真空除去溶剂，将残余物用饱和NaHCO 3水溶液(30mL)稀释，然后用二氯甲烷(30mL×2)萃取。合并的有机相用盐水(30mL)洗涤，经硫酸钠干燥，并过滤。将滤液在减压下浓缩。粗 残余物通过快速柱色谱法纯化(0-3％甲醇的二氯甲烷溶液)，得到标题化合物(400mg，38％)，为无色油状物。LC-MS:ESI m/z:523.2[M+H] +；C 30H 42N 4O 4计算值522.32.
步骤5：2-(甲基((S)-1-(3-(5,6,7,8-四氢-1,8-萘啶-2-基)丙基)吡咯烷-3-基)氨基)-2-苯基乙酸
Figure PCTCN2021078537-appb-000105
将7-(3-((3S)-3-((2-甲氧基-2-氧-1-苯基乙基)(甲基)氨基)吡咯烷-1-基)丙基)-3,4-二氢-1，8-萘啶-1(2H)-羧酸叔丁酯(400mg，0.76mmol)溶解在盐酸/1，4-二氧六环(10mL，4N，40mmol)溶液中。所得溶液在25℃下搅拌3小时后，真空除去溶剂。将残余物溶于甲醇(10mL)和H 2O(2.5mL)的混合溶剂中，然后加入LiOH(91mg，3.8mmol)。将混合物在40℃下搅拌4小时，然后用10％柠檬酸水溶液中和至pH≈7。真空浓缩溶剂。将残余物通过制备型反相HPLC在以下条件下纯化[柱：Kromasil Prep C18，30×150mm；流动相：含0.1％甲酸的1-40％乙腈水溶液，15分钟]，得到2-(甲基((S)-1-(3-(5,6,7,8-四氢-1,8-萘啶-2-基)丙基)吡咯烷-3-基)氨基)-2-苯基乙酸(71mg，23％)的差向异构体混合物。然后通过制备手性SFC，在以下条件下分离非对映异构体混合物[柱：Chiralpak AD-3(50×4.6mm)；流动相：40％乙醇的二氧化碳溶液(含0.05％DEA)；流速：3mL/min]，分别得到化合物5a和化合物5b，为白色固体。
化合物5a: 1H NMR(400MHz,CD 3OD)δ7.51–7.49(m,2H),7.42–7.37(m,4H),6.50(d,J＝7.3Hz,1H),4.48(s,1H),3.97–3.84(m,1H),3.45–3.36(m,2H),3.29–3.16(m,3H),2.87–2.60(m,7H),2.41(s,3H),2.23–2.18(m,2H),2.02–1.73(m,4H).LC-MS:RT＝0.68min,100％,ESI m/z:409.1[M+H] +；C 24H 32N 4O 2计算值408.25.采用分析型手性SFC(ChiralPak AD-3(50×4.6mm)柱子)，保留时间：2.16min，ee值：100.0％.
化合物5b： 1H NMR(400MHz,CD 3OD)δ7.52–7.49(m,2H),7.39–7.32(m,3H),7.16(d,J＝7.3Hz,1H),6.39(d,J＝7.3Hz,1H),4.27(s,1H),3.79–3.71(m,1H),3.31–3.28(m,2H),3.20–3.14(m,3H),2.78–2.61(m,7H),2.49(s,3H),2.14–2.03(m,2H),1.92–1.60(m,4H).LC-MS:ESI m/z:409.3[M+H] +；C 24H 32N 4O 2计算值408.25.采用分析型手性SFC(ChiralPak AD-3(50×4.6mm)柱子)，保留时间：2.86min，ee值：97.7％.
实施例4 可参照化合物1、化合物5的合成方法，得到以下化合物：
化合物6(甲酸盐)： 1H NMR(400MHz,CD 3OD)δ7.58–7.49(m,2H),7.48–7.36(m,4H),6.53(dd,J＝7.3,2.7Hz,1H),4.62–4.51(m,1H),3.91–3.81(m,1H),3.46–3.37(m,3H),3.28–3.18(m,2H),2.81–2.67(m,6H),2.62–2.49(m,4H),2.27–2.11(m,2H),1.95–1.85(m,4H).LC-MS:ESI m/z 409.1[M+H] +；C 24H 32N 4O 2计算值408.25.
化合物7(甲酸盐)： 1H NMR(400MHz,CD 3OD)δ7.58–7.29(m,6H),6.61–6.55(m,1H),4.65(s,0.67H),4.53(s,0.33H),3.82–3.80(m,1H),3.58–3.40(m,4H),3.33–3.31(m,1H),3.20–3.00(m,3H),2.78–2.72(m,4H),2.50(s,2H),2.38(s,1H),2.22–2.12(m,2H),1.98–1.71(m,6H).LC-MS:ESI m/z 423.3[M+H] +；C 25H 34N 4O 2计算值422.27.
化合物8(甲酸盐)： 1H NMR(400MHz,CD 3OD)δ7.56–7.43(m,3H),7.43–7.28(m,3H),6.55(d,J＝7.3Hz,1H),4.49(d,J＝28.0Hz,1H),3.88–3.75(m,1H),3.49–3.42(m,2H),3.42–3.31 (m,2H),3.17–3.09(m,1H),3.07–2.91(m,3H),2.79(t,J＝6.2Hz,2H),2.67(t,J＝7.8Hz,2H),2.40(d,J＝40.0Hz,3H),2.19(dd,J＝14.5,7.3Hz,1H),2.12(dd,J＝14.8,7.4Hz,1H),1.97–1.88(m,2H),1.80–1.61(m,4H),1.52–1.40(m,2H).LC-MS:ESI m/z 437.24[M+H] +,C 26H 36N 4O 2.计算值436.28.
化合物9(甲酸盐)： 1H NMR(400MHz,CD 3OD)δ7.51–7.42(m,6H),6.58–6.57(m,1H),4.61–4.60(m,1H),3.43–3.31(m,4H),3.04–2.75(m,8H),2.55–2.48(m,3H),223–1.94(m,3H),1.92–1.86(m,2H).LC-MS:ESI m/z 395.1[M+H] +；C 23H 30N 4O 2计算值394.24.
制备得到化合物10，通过制备手性SFC，在以下条件下分离非对映混合物：[色谱柱：Chiralpak AD-3(50×4.6mm)，流动相：40％乙醇的CO 2溶液(含0.05％DEA)，流速：3mL/min]得到化合物10a和化合物10b，分别为白色固体。
化合物10a： 1H NMR(400MHz,CD 3OD)δ7.51(d,J＝6.5Hz,2H),7.36–7.31(m,3H),7.13(d,J＝7.2Hz,1H),6.37(d,J＝7.3Hz,1H),4.29(s,1H),3.79–3.67(m,1H),3.37–3.34(m,2H),3.24–2.94(m,3H),2.86–2.66(m,5H),2.59(t,J＝7.0Hz,2H),2.45(s,3H),2.13–2.01(m,2H),1.97–1.74(m,4H).LC-MS ESI m/z:409.1[M+H] +；C 24H 32N 4O 2计算值408.25.采用分析型手性SFC(ChiralPak IG-3(50×4.6mm)柱子)，保留时间：2.07min，ee值：100.0％.
化合物10b： 1H NMR(400MHz,CD 3OD)δ7.52(dd,J＝7.7,1.6Hz,2H),7.38–7.33(m,3H),7.14(d,J＝7.3Hz,1H),6.37(d,J＝7.3Hz,1H),4.29(s,1H),3.91–3.83(m,1H),3.39–3.34(m,2H),3.22–2.98(m,3H),2.96–2.64(m,5H),2.59(t,J＝7.3Hz,2H),2.39(s,3H),2.16–2.08(m,2H),1.98–1.79(m,4H).LC-MS ESI m/z:409.1[M+H] +,C 24H 32N 4O 2计算值408.25.采用分析型手性SFC(ChiralPak AD-3(50×4.6mm)柱子)，保留时间：3.88min，ee值：100.0％.
制备得到化合物11，通过制备手性SFC，在以下条件下分离非对映异构体混合物：[柱：(S，S)Whelk-O1(100×4.6mm)；流动相：40％甲醇的二氧化碳溶液(含0.05％DEA)；流速：3mL/min]，分别得到化合物11a和化合物11b，为白色固体。
化合物11a： 1H NMR(400MHz,CD 3OD)δ7.42(d,J＝7.3Hz,2H),7.26–7.23(m,3H),7.01(d,J＝7.4Hz,1H),6.25(d,J＝7.3Hz,1H),4.14(s,1H),3.83–3.73(m,1H),3.30–3.22(m,2H),3.04–2.93(m,2H),2.87–2.64(m,4H),2.60–2.51(m,2H),2.41(t,J＝7.4Hz,2H),2.23(s,3H),2.02–1.96(m,2H),1.82–1.68(m,2H),1.61–1.38(m,4H),1.30–1.19(m,2H).LC-MS:RT＝0.35min,100％,ESI m/z:437.2[M+H] +；C 26H 36N 4O 2计算值436.28.采用分析型手性SFC((S,S)Whelk-O1(100×4.6mm)柱子)得到保留时间：3.21min，ee值：100.0％.
化合物11b： 1H NMR(400MHz,CD 3OD)δ7.53(d,J＝6.8Hz,2H),7.39–7.35(m,3H),7.19(d,J＝6.9Hz,1H),6.40(d,J＝7.0Hz,1H),4.34(s,1H),3.81–3.69(m,1H),3.43–3.37(m,2H),3.24–3.13(m,2H),3.04–2.77(m,4H),2.76–2.66(m,2H),2.62–2.50(m,2H),2.47(s,3H),2.15–2.02(m,2H),1.94–1.82(m,2H),1.76–1.54(m,4H),1.47–1.27(m,2H).LC-MS:ESI m/z:437.2[M+H] +；C 26H 36N 4O 2计算值436.28.采用分析型手性SFC((S,S)Whelk-O1(100×4.6mm)柱子)得到保留时间：4.43min，ee值：96.6％.
制备得到化合物12，通过制备手性SFC，在以下条件下分离非对映异构体混合物[柱：Chiralpak IC-3(50×4.6mm)；流动相：二氧化碳中的40％甲醇(含0.05％DEA)；流速：3mL/min]，分别得到化合物12a和化合物12b，为白色固体。
化合物12a： 1H NMR(400MHz,CD 3OD)δ7.63–7.31(m,6H),6.46(d,J＝6.6Hz,1H),4.40(s,1H),3.89–3.75(m,1H),3.52–3.38(m,2H),3.26–2.70(m,8H),2.69–2.57(m,2H),2.48(s,3H),2.22–2.03(m,2H),1.98–1.85(m,2H),1.84–1.53(m,4H),1.49–1.32(m,2H).LC-MS:ESI m/z:437.4[M+H] +；C 26H 36N 4O 2计算值436.28.采用分析型手性SFC(Chiralpak IC-3(50×4.6mm)柱子)得到保留时间：1.53min，ee值：97.8％.
化合物12b： 1H NMR(400MHz,CD 3OD)δ7.49–7.38(m,2H),7.34–7.17(m,3H),7.06(d,J＝7.1Hz,1H),6.27(d,J＝7.3Hz,1H),4.19(s,1H),3.86–3.77(m,1H),3.31–3.24(m,2H),3.13–2.99(m,2H),2.95–2.67(m,4H),2.62–2.53(m,2H),2.48–2.37(m,2H),2.24(s,3H),2.05–1.98(m,2H),1.83–1.71(m,2H),1.54–1.46(m,4H),1.34–1.24(m,2H).LC-MS:ESI m/z:437.4[M+H] +；C 26H 36N 4O 2计算值436.28.采用分析型手性SFC(Chiralpak IC-3(50×4.6mm)柱子)得到保留时间：2.13min，ee值：100.0％.
制备得到化合物13，通过制备手性SFC，在以下条件下分离粗残留物[柱：Diacel ChiralPak IG(250×30mm，10μm)。流动相：60％甲醇的二氧化碳溶液(含0.1％NH 3.H 2O)；流速：70g/min]，分别得到化合物13a和化合物13b，为浅黄色固体。
化合物13a： 1H NMR(400MHz,CD 3OD)δ7.41(d,J＝6.1Hz,2H),7.33–7.26(m,4H),6.40(d,J＝7.3Hz,1H),4.28(s,1H),3.82(br,1H),3.37–3.30(m,2H),3.13–2.82(m,6H),2.66(t,J＝6.1Hz,2H),2.56(t,J＝7.1Hz,2H),2.27(s,3H),2.09–2.07(m,2H),1.84–1.78(m,2H),1.62–1.59(m,4H).LC-MS:ESI m/z 423.2[M+H] +；C 25H 34N 4O 2计算值422.27.采用分析型手性SFC(Chiralpak IG(250×30mm)柱子)得到保留时间：2.713min，ee值：100.0％.
化合物13b： 1H NMR(400MHz,CD 3OD)δ7.52(d,J＝6.8Hz,2H),7.37–7.35(m,3H),7.25–7.21(m,1H),6.45–6.40(m,1H),4.38–4.32(m,1H),3.76(br,1H),3.45–3.36(m,2H),3.21(br,2H),3.00–2.89(m,4H),2.72(t,J＝5.7Hz,2H),2.58(br,2H),2.45(s,3H),2.08(br,2H),1.96–1.84(m,2H),1.67(br,4H).LC-MS:ESI m/z 423.2[M+H] +；C 25H 34N 4O 2计算值422.27.采用分析型手性SFC(Chiralpak IG(250×30mm)柱子)得到保留时间：3.364min，ee值：94.1％.
制备得到化合物14，通过制备手性SFC，在以下条件下分离粗残余物：[柱：Daicel ChiralPak IG(250×30mm，10μm)；柱状色谱。流动相：60％甲醇的二氧化碳溶液(含0.1％NH 3.H 2O)流速：70g/min]，分别得到化合物14a和化合物14b，为浅黄色固体。
化合物14a： 1H NMR(400MHz,CD 3OD)δ7.41(d,J＝6.8Hz,2H),7.28–7.18(m,3H),7.03(d,J＝8.0Hz,1H),6.26(d,J＝7.3Hz,1H),4.16(s,1H),3.66–3.63(m,1H),3.31–3.23(m,2H),3.10–3.03(m,2H),2.92–2.81(m,1H),2.80–2.66(m,3H),2.59(t,J＝6.2Hz,2H),2.45(t,J＝6.4Hz,2H),2.32(s,3H),1.99–1.94(m,2H),1.84–1.72(m,2H),1.65–1.46(m,4H).LC-MS:ESI m/z 423.2[M+H] +；C 25H 34N 4O 2计算值422.27.采用分析型手性SFC(Chiralpak IG(250×30mm)柱子)得到保留时间：2.870min，ee值：100.0％.
化合物14b： 1H NMR(400MHz,CD 3OD)δ7.53(d,J＝6.2Hz,2H),7.44–7.31(m,3H),7.19–7.11(m,1H),6.39–6.35(m,1H),4.30(s,1H),3.92–3.89(m,1H),3.43–3.35(m,2H),3.21–3.15(m,2H),3.02–2.98(m,1H),2.91–2.78(m,3H),2.77–2.67(m,2H),2.64–2.52(m,2H),2.37(s,3H),2.21–2.10(m,2H),1.96–1.84(m,2H),1.77–1.57(m,4H).LC-MS:ESI m/z 423.2[M+H] +；C 25H 34N 4O 2计算值422.27.采用分析型手性SFC(Chiralpak IG(250×30mm)柱子)得到保留时间：4.351min，ee值：97.0％.
实施例5 化合物15的合成
Figure PCTCN2021078537-appb-000106
步骤1：7-(3-(3-(((R)-2-甲氧基-2-氧-1-苯基乙基)(甲基)氨基)吡咯烷-1-基)-3-氧丙基)-3，4-二氢-1,8-萘啶-1(2H)-羧酸叔丁酯
Figure PCTCN2021078537-appb-000107
在N 2气氛下，向(2R)-2-(甲基(吡咯烷基-3-基)氨基)-2-苯基乙酸甲酯(60mg，0.24mmol)和3-(8-(叔丁氧羰基)-5,6,7,8-四氢-1,8-萘啶-2-基)丙酸(88mg，0.29mmol)的DMF(2mL)溶液中加入HBTU(136mg，0.36mmol)和三乙胺(0.15mL，1.2mmol)。在25℃下搅拌16小时后，将反应混合物用饱和NaHCO 3(15mL)稀释，并用乙酸乙酯(15mL×2)萃取。合并的有机相用盐水洗涤，经硫酸钠干燥，过滤并浓缩至干。残余物通过快速柱色谱法纯化(0-8％甲醇的二氯甲烷溶液)，得到标题化合物(38mg，56％)，为黄色油状物。LC-MS:ESI m/z 537.2[M+H] +；C 30H 40N 4O 5计算值536.30.
步骤2：(2R)-2-(甲基(1-(3-(5,6,7,8-四氢-1,8-萘啶-2-基)丙酰基)吡咯烷-3-基)氨基)-2-苯基乙酸甲酯
Figure PCTCN2021078537-appb-000108
向7-(3-(3-(((R)-2-甲氧基-2-氧-1-苯基乙基)(甲基)氨基)吡咯烷-1-基)-3-氧丙基)-3,4-二氢-1,8-萘啶-1(2H)-羧酸叔丁酯(38mg，0.07mmol)的二氯甲烷(1mL)溶液中加入TFA(1mL)，并将所得混合物在25℃下搅拌16小时。真空除去溶剂，得到标题化合物(30mg，99％)，为浅黄色油状的TFA盐形式，其无需进一步纯化直接用于下一步。LC-MS:ESI m/z 437.2[M+H] +；C 25H 32N 4O 3.计算值436.25.
步骤3：(2R)-2-(甲基(1-(3-(5,6,7,8-四氢-1,8-萘啶-2-基)丙酰基)吡咯烷-3-基)氨基)-2-苯乙酸
Figure PCTCN2021078537-appb-000109
向TFA盐形式的(2R)-2-(甲基(1-(3-(5,6,7,8-四氢-1,8-萘啶-2-基)丙酰基)吡咯烷-3-基)氨基)-2-苯基乙酸甲酯(30mg，0.07mmol)的甲醇(2mL)和水(0.5mL)溶液中加入NaOH(5.6mg，0.14mmol)，并将得到的混合物在25℃下搅拌16小时。用1N盐酸(0.2mL)中和后，将混合物浓缩并将残余物通过制备HPLC在以下条件下纯化[柱：Kromasil 100-5-C18，30×150mm；流动相：1-100％乙腈的水溶液(含0.1％甲酸)，14分钟]，得到标题化合物(26.9mg，49％)，为白色固体。 1H NMR(400MHz,CD 3OD)δ7.55–7.53(m,2H),7.46–7.37(m,4H),6.57–6.49(m,1H),4.64–4.50(m,1H),4.00–3.74(m,2H),3.74–3.31(m,6H),2.99–2.88(m,2H),2.78–2.70(m,3H),2.51–2.43(m,3H),2.35–2.00(m,2H),1.94–1.85(m,2H).LC-MS:ESI m/z 423.14[M+H] +；C 24H 30N 4O 3计算值422.23.
实施例6.参照化合物15的合成方法，并结合拆分手段，得到以下化合物：
制备得到化合物16，将混合物浓缩并将残余物通过制备HPLC在以下条件下纯化[柱：Kromasil 100-5-C18，30×150mm；流动相：1-100％乙腈的水溶液(含0.1％甲酸)，14分钟]，分别得到化合物16a和化合物16b。
化合物16a： 1H NMR(400MHz,CD 3OD)δ7.58–7.24(m,6H),6.53–6.45(m,1H),4.49–4.45(m,1H),3.73–3.58(m,3H),3.50–3.37(m,4H),2.90–2.82(m,2H),2.77–2.58(m,4H),2.25–1.94(m,2H),1.92–1.85(m,2H).LC-MS:ESI m/z 409.1[M+H] +；C 23H 28N 4O 3计算值408.22.采用分析型手性SFC(Chiralpak IG(250×30mm)柱子)得到保留时间：2.713min，ee值：100.0％.
化合物16b： 1H NMR(400MHz,CD 3OD)δ7.61–7.32(m,6H),6.61–6.55(m,1H),4.60–4.52(m,1H),4.00–3.90(m,2H),3.75–3.68(m,2H),3.62–3.49(m,1H),3.45–3.39(m,2H),3.04–2.93(m,2H),2.85–2.61(m,3H),2.48–2.39(m,1H),2.25–1.98(m,2H),1.97–1.88(m,2H).LC-MS:ESI m/z 409.1[M+H] +,C 23H 28N 4O 3.计算值408.22.
化合物17(甲酸盐)： 1H NMR(400MHz,CD 3OD)δ7.54–7.52(m,2H),7.45–7.37(m,4H),6.54–6.52(m,1H),4.54–4.51(m,1H),3.70–3.65(m,2H),3.57–3.53(m,1H),3.45–3.42(m,2H),2.78–2.77(m,2H),2.67–2.64(m,2H),2.43–2.02(m,6H),1.94–1.88(m,2H),1.68–1.63(m,4H).LC-MS:ESI m/z 437.2[M+H] +；C 25H 32N 4O 3计算值436.25.
化合物18(甲酸盐)： 1H NMR(400MHz,CD 3OD)δ7.53–7.40(m,6H),6.57–6.52(m,1H),4.61–4.54(m,1H),3.93–3.40(m,8H),2.79–2.75(m,2H),2.71–2.65(m,2H),2.47–2.34(m,6H),1.94–1.88(m,2H),1.75–1.66(m,4H).LC-MS:ESI m/z 451.1[M+H] +；C 26H 34N 4O 3计算值450.26.
化合物19(甲酸盐)： 1H NMR(400MHz,CD 3OD)δ7.61–7.52(m,3H),7.50–7.39(m,3H),6.65–6.57(m,1H),4.69–4.62(m,1H),3.96–3.62(m,6H),3.48–3.42(m,2H),2.81–2.80(m,2H),2.48–2.00(m,3H),1.97–1.89(m,2H).LC-MS:ESI m/z 395.1[M+H] +；C 22H 26N 4O 3计算值394.20.
化合物20(甲酸盐)： 1H NMR(400MHz,CD 3OD)δ7.56–7.42(m,5H),7.37–7.33(m,1H)6.53–6.48(m,1H),4.62–4.56(m,1H),4.01–3.36(m,8H),2.78–2.74(m,2H),2.57–2.49(m,3H),2.38–2.02(m,3H),1.93–1.86(m,2H).LC-MS:ESI m/z 409.1[M+H] +；C 23H 28N 4O 3计算值 408.22.
实施例7.化合物21的合成
Figure PCTCN2021078537-appb-000110
步骤1：3-(苯基氨基)吡咯烷-1-羧酸叔丁酯
Figure PCTCN2021078537-appb-000111
在25℃下，将3-氧吡咯烷-1-羧酸叔丁酯(6.0g，32.4mmol)和苯胺(3.0g，32.4mmol)溶解在四氢呋喃(100mL)和冰醋酸(25mL)的混合溶液，然后加入BH 3·THF(1N，120mL)。将反应混合物在25℃搅拌2小时，真空浓缩以除去四氢呋喃，将反应混合物用饱和NaHCO 3(30mL)稀释，并用乙酸乙酯(30mL×2)萃取。将合并的有机相用水和盐水洗涤，经硫酸钠干燥，并在减压下浓缩，得到的粗产物通过柱色谱法纯化(0～20％乙酸乙酯的石油醚溶液)，得到标题化合物(3.0g，35％产率)，为白色固体。LC-MS:ESI m/z 363.2[M+H] +；C 15H 22N 2O 2计算值262.17.
步骤2：3-((2-甲氧基-2-氧乙基)(苯基)氨基)吡咯烷-1-羧酸叔丁酯
Figure PCTCN2021078537-appb-000112
向3-(苯氨基)吡咯烷-1-羧酸叔丁酯(1.5g，5.7mmol)的乙腈(30mL)溶液，加入KHCO 3(1.7g，17.1mmol)，NaI(855mg，5.7mmol)和2-溴乙酸甲酯(1.7g，11.4mmol)。将反应混合物在80℃下搅拌30小时。将反应混合物用饱和NaHCO 3(30mL)稀释，并用乙酸乙酯(30mL×2)萃取。合并的有机相用盐水洗涤，经硫酸钠干燥并在减压下浓缩，得到的粗产物通过柱色谱法纯化(0～20％乙酸乙酯的石油醚溶液)，得到黄色油状的标题化合物(1.9g，98％产率)。LC-MS:ESI m/z 335.3[M+H] +；C 18H 26N 2O 4计算值334.19.
步骤3：N-苯基-N-(吡咯烷-3-基)甘氨酸甲酯
Figure PCTCN2021078537-appb-000113
向3-((2-甲氧基-2-氧乙基)(苯基)氨基)吡咯烷-1-羧酸叔丁酯(1.9g，5.7mmol)的二氯甲烷(10mL)溶液中加入盐酸/1，4-二氧六环溶液(4N，5mL)。将反应混合物在25℃下搅拌15小时。减压浓缩所得混合物，得到标题化合物(1.6g，粗品)，为黄色固体。LC-MS:ESI m/z 235.2[M+H] +；C 13H 18N 2O 2计算值234.14.
步骤4：7-(3-(3-((2-甲氧基-2-氧乙基)(苯基)氨基)吡咯烷-1-基)丙基)-3,4-二氢-1,8-萘啶-1(2H)-羧酸叔丁酯
Figure PCTCN2021078537-appb-000114
向N-苯基-N-(吡咯烷基-3-基)甘氨酸甲酯(200mg，0.85mmol)的1,2-二氯乙烷(5mL)的溶液中，加入氰基硼氢化钠(213mg，3.40mmol)，反应混合物在25℃搅拌2小时。将反应混合物用饱和NaHCO 3(15mL)稀释，并用二氯甲烷(15mL×2)萃取。合并的有机相用盐水洗涤，经硫酸钠干燥并在减压下浓缩以得到粗产物，将其通过柱色谱法纯化(0～10％甲醇的二氯甲烷溶液)，得到标题化合物(230mg，53％产率)，为黄色固体。LC-MS:ESI m/z 509.4[M+H] +；C 29H 40N 4O 4计算值508.30.
步骤5：N-苯基-N-(1-(3-(5,6,7,8-四氢-1,8-萘啶-2-基)丙基)吡咯烷基-3-基)甘氨酸甲酯
Figure PCTCN2021078537-appb-000115
向7-(3-(3-((2-甲氧基-2-氧乙基)(苯基)氨基)吡咯烷-1-基)丙基)-3,4-二氢-1,8-萘啶-1(2H)-羧酸叔丁酯(230mg，0.45mmol)的二氯甲烷(2mL)溶液中加入盐酸/1，4-二氧六环(3mL，12mmol)溶液。将反应混合物在25℃下搅拌15小时。真空减压浓缩，得到标题化合物(140mg，76％产率)，为白色固体。LC-MS:ESI m/z 409.3[M+H] +；C 24H 32N 4O 2计算值408.25.
步骤6：N-苯基-N-(1-(3-(5,6,7,8-四氢-1,8-萘啶-2-基)丙基)吡咯烷-3-基)甘氨酸
Figure PCTCN2021078537-appb-000116
向N-苯基-N-(1-(3-(5,6,7,8-四氢-1,8-萘啶-2-基)丙基)吡咯烷基-3-基)甘氨酸甲酯(130mg，0.32mmol)的甲醇(3mL)溶液中加入NaOH(4N，2mL)。将反应混合物在25℃下搅拌15小时。将反应混合物减压浓缩以除去甲醇，并将所得混合物用1N盐酸酸化至pH＝2～3。滤出粗产物，将其用制备HPLC纯化(柱：Gilson C18，流动相：50-100％乙腈的水溶液，含0.1％甲酸)，得到标题化合物(20mg，16％产率)，为白色固体。 1H NMR(400MHz,DMSO-d 6)δ7.20–7.09(m,2H),7.03(d,J＝7.2Hz,1H),6.70–6.61(m,3H),6.35(s,1H),6.26(d,J＝7.2Hz,1H),4.50–4.44(m,1H),4.14(d,J＝17.9Hz,1H),3.82(d,J＝17.9Hz,1H),3.26–3.21(m,2H), 3.14–3.08(m,1H),3.02–2.96(m,1H),2.65–2.52(m,5H),2.48–2.44(m,2H),2.41–2.36(m,1H),2.33–2.26(m,1H),1.86–1.71(m,5H).LC-MS:ESI m/z 395.2[M+H] +；C 23H 30N 4O 2计算值394.24.
实施例8.化合物22的合成
Figure PCTCN2021078537-appb-000117
步骤1：3-(苄基(2-乙氧基-2-氧乙基)氨基)吡咯烷-1-羧酸叔丁酯
Figure PCTCN2021078537-appb-000118
在N 2气氛下，向苄基甘氨酸乙酯(2g，10.34mmol)的3-氧杂吡咯烷-1-甲酸叔丁酯(1.9g，10.34mmol)的甲醇(20mL)溶液中添加三乙酰氧基硼氢化钠(6.5g，31.02mmol)，在40℃下搅拌15小时后，加入氰基硼氢化钠(1.9g，31.02mmol)，在40℃下再搅拌15小时。将反应混合物用水(20mL)淬灭，用乙酸乙酯(20mL×2)萃取。合并的有机相用盐水洗涤，经硫酸钠干燥并在减压下浓缩，得到粗产物通过柱色谱法纯化(0～30％乙酸乙酯的石油醚溶液)，得到无色油状的标题化合物(1.7g，45％产率)。
步骤2～5，参照化合物21的合成步骤3～6的相似条件，得到化合物22(甲酸盐)(20mg，27％产率)，为白色固体。 1H NMR(400MHz,CD 3OD)δ7.48(d,J＝7.3Hz,1H),7.44–7.39(m,2H),7.36–7.30(m,2H),7.29–7.23(m,1H),6.56(d,J＝7.3Hz,1H),4.07(d,J＝14.2Hz,1H),3.78–3.63(m,3H),3.61–3.55(m,1H),3.44–3.36(m,3H),3.32–3.31(m,1H),3.18–3.06(m,4H),2.88–2.73(m,4H),2.34–2.24(m,2H),2.18–2.07(m,1H),2.00–1.87(m,3H).
实施例9.化合物23的合成
Figure PCTCN2021078537-appb-000119
步骤1：(3R)-3-((1-(3-氯苯基)-3-甲氧基-3-氧丙基)氨基)吡咯烷-1-羧酸叔丁酯
Figure PCTCN2021078537-appb-000120
将3-(3-氯苯基)-3-氧代丙酸甲酯(2.0g，9.4mmol)，(R)-3-氨基吡咯烷-1-羧酸叔丁酯(2.6g，14mmol)和三乙酰氧基硼氢化钠(3.99g，19mmol)在甲醇(30mL)和冰醋酸(3.0mL)的混合物于25℃搅拌12小时。然后加入氰基硼氢化钠(1.8g，19mmol)，在25℃下再搅拌24小时。将混合物浓缩至干，并添加水(20mL)稀释，用二氯甲烷(30mL×3)萃取。合并的有机相用NaHCO 3(50mL)洗涤，经硫酸钠干燥并浓缩至干。粗产物通过FCC纯化(20～100％乙酸乙酯的石油醚溶液)，得到黄色胶状的标题化合物(1.2g，33％)。 1H NMR(400MHz,CDCl 3)δ7.37–7.19(m,4H),4.15–4.02(m,1H),3.67–3.66(m,3H),3.55–3.30(m,2H),3.29–3.14(m,1H),3.13–2.89(m,2H),2.68–2.53(m,2H),2.02–1.83(m,1H),1.82–1.55(m,1H),1.51–1.37(m,9H).LC-MS:ESI m/z 383.2,385.2[M+H,Cl] +；C 19H 27ClN 2O 4计算值382.17.
步骤2：3-(3-氯苯基)-3-(((R)-吡咯烷基-3-基)氨基)丙酸甲酯
Figure PCTCN2021078537-appb-000121
在25℃下，将(3R)-3-((1-(3-氯苯基)-3-甲氧基-3-氧丙基)氨基)吡咯烷-1-羧酸叔丁酯(0.60g，0.6mmol)的二氯甲烷(2mL)和盐酸/1，4-二氧六环溶液(5.9mL，23mmol)的混合物搅拌14小时。将混合物浓缩至干，得到标题化合物(0.52g，粗品)，其无需进一步纯化直接用于下一步。LC-MS:ESI m/z 283.0,285.2[M+H,Cl] +；C 14H 19ClN 2O 2计算值282.11.
步骤3：7-(2-((3R)-3-((1-(3-氯苯基)-3-甲氧基-3-氧丙基)氨基)吡咯烷-1-基)乙基)-3,4-二氢-1,8-萘啶-1(2H)-羧酸叔丁酯
Figure PCTCN2021078537-appb-000122
在25℃，N 2气氛下，向3-(3-氯苯基)-3-(((R)-吡咯烷基-3-基)氨基)丙酸甲酯(0.25g，1.3mmol)和7-(2-碘乙基)-3,4-二氢-1,8-萘啶-1(2H)-羧酸叔丁酯(0.52g，1.3mmol)的乙腈(10mL)溶液中分三批加入DIPEA(0.23g，1.8mmol)。将反应混合物在25℃下搅拌18小时。将该混合物倒入水(50mL)中，并用乙酸乙酯(30mL×2)萃取。合并的有机相用盐水洗涤，经硫酸钠干燥，过滤并浓缩，得到粗产物，将其通过硅胶柱色谱法(1～3.4％ 甲醇的二氯甲烷溶液)纯化，得到标题化合物(0.35g，52％)，为黄色油状物。LC-MS:ESI m/z 543.3,545.3[M+H,Cl] +；C 29H 39ClN 4O 4计算值542.27.
步骤4：3-(3-氯苯基)-3-(((R)-1-(2-(5,6,7,8-四氢-1,8-萘啶-2-基)乙基)吡咯烷酮-3-基)氨基)丙酸甲酯
Figure PCTCN2021078537-appb-000123
在N 2气氛下，向7-(2-((3R)-3-((1-(3-氯苯基)-3-甲氧基-3-氧丙基)氨基)吡咯烷-1-基)乙基)-3,4-二氢-1,8-萘啶-1(2H)-羧酸叔丁酯(0.30g，0.55mmol)的甲醇(5mL)溶液中加入盐酸/1，4-二氧六环(0.82mL，3.2mmol)，并在25℃下搅拌18小时。将混合物减压浓缩，得到红色固体的标题化合物(0.26g，粗品)，其无需进一步纯化直接用于下一步。LC-MS:ESI m/z 443.3,445.3[M+H,Cl] +；C 24H 31ClN 4O 2计算值442.21.
步骤5：3-(3-氯苯基)-3-(((R)-1-(2-(5,6,7,8-四氢-1,8-萘啶-2-基)乙基)吡咯烷酮3-基)氨基)丙酸
Figure PCTCN2021078537-appb-000124
向3-(3-氯苯基)-3-(((R)-1-(2-(5,6,7,8-四氢-1,8-萘啶-2-基)乙基)吡咯烷-3-基)氨基)丙酸甲酯(0.26g)的四氢呋喃(5mL)，水(3mL)和甲醇(5mL)溶液中分三批加入LiOH·H 2O(98mg，2.3mmol)，并在25℃下搅拌2小时。将混合物用盐酸(1N)酸化至pH＝4～5，并浓缩至干。粗产物通过制备型HPLC(柱：Xbridge 5u C18 150×19mm；流动相：乙腈-水(0.05％甲酸)；梯度：5～15％乙腈；流速：20mL/min)纯化，得到标题化合物(0.12g，32％)，为黄色固体。 1H NMR(400MHz,DMSO-d 6)δ7.42(s,1H),7.32–7.26(m,3H),7.02(dd,J＝10.8,7.2Hz,1H),6.27(dd,J＝14.4,7.2Hz,1H),3.99–3.96(m,1H),3.24–3.22(m,2H),3.02–2.98(m,1H),2.81–2.49(m,10H),2.39–2.23(m,2H),1.96–1.81(m,1H),1.81–1.70(m,2H),1.65–1.38(m,1H).LC-MS:ESI m/z 429.2,431.2[M+H,Cl] +；C 23H 29ClN 4O 2计算值428.20.HPLC纯度:97.2％(214nm),99.1％(254nm).
实施例10.
参照化合物23的合成方法，采用
Figure PCTCN2021078537-appb-000125
替代相应原料制备得到化合物24，混合物通过制备HPLC纯化(色谱柱：Kromasil-C18 100×21.2mm 5um；流动相：乙腈-水(0.1％甲酸)；梯度：10～20％)，得到化合物24a和化合物24b。
化合物24a: 1H NMR(400MHz,DMSO-d 6)δ7.47(s,1H),7.40–7.30(m,3H),7.05(d,J＝7.2Hz,1H),6.39(brs,1H),6.28(d,J＝7.3Hz,1H),4.04–4.00(m,1H),3.26–3.22(m,2H),3.03–3.01(m,1H),2.97–2.82(m,5H),2.73–2.52(m,6H),2.49–2.31(m,1H),2.00–1.90(m,1H),1.77–1.68(m,3H)；LC-MS:ESI m/z 429.2,431.2[M+H,Cl] +；C 23H 29ClN 4O 2计算值428.20.HPLC纯度:98.0％(214nm),97.5％(254nm).
化合物24b: 1H NMR(400MHz,DMSO-d 6)δ7.48(s,1H),7.39–7.29(m,3H),7.09(d,J＝7.2Hz,1H),6.62(brs,1H),6.32(d,J＝7.2Hz,1H),3.99–3.97(m,1H),3.28–3.22(m,2H),3.04–2.81(m,7H),2.73–2.71(m,2H),2.63–2.52(m,3H),2.40–2.32(m,1H),2.00–1.90(m,1H),1.79–1.72(m,2H),1.55–1.50(m,1H).LC-MS:ESI m/z 429.3,431.3[M+H,Cl] +；C 23H 29ClN 4O 2计算值428.20.HPLC纯度:99.1％(214nm),98.8％(254nm).
参照化合物23的合成方法，制备得到以下化合物：
化合物25(甲酸盐)： 1H NMR(400MHz,DMSO-d 6)δ7.63–7.61(m,2H),7.60–7.54(m,2H),7.08–7.04(m,1H),6.33–6.26(m,1H),4.15–4.06(m,1H),3.26–3.22(m,2H),3.09–2.87(m,6H),2.71–2.61(m,5H),2.46–2.39(m,2H),1.98–1.81(m,1H),1.73-1.71(m,2H),1.67–1.39(m,1H).LC-MS:ESI m/z 463.3[M+H] +；C 24H 29F 3N 4O 2计算值462.22.HPLC纯度:97.0％(214nm),97.3％(254nm).
化合物26(甲酸盐)： 1H NMR(400MHz,DMSO-d 6)δ7.37–7.26(m,4H),7.22–7.18(m,1H),7.00–6.98(m,1H),6.43(brs,1H),6.28–6.23(m,1H),3.93–3.87(m,1H),3.24–3.19(m,2H),2.91–2.82(m,1H),2.66–2.52(m,6H),2.49–2.24(m,4H),2.20–1.95(m,2H),1.85–1.70(m,3H),1.69–1.24(m,1H).LC-MS:ESI m/z 395.3[M+H] +；C 23H 30N 4O 2计算值394.24.HPLC纯度:100.0％(214nm),99.7％(254nm).
制备得到化合物27，并通过制备型HPLC(色谱柱：Kromasil-C18 100×21.2mm 5um；流动相：乙腈-水(0.1％甲酸)；梯度：10～20％)纯化粗品，得到化合物27a和化合物27b。
化合物27a： 1H NMR(400MHz,DMSO-d 6,80℃)δ8.06(s,1H),7.98(s,1H),7.67(s,2H),7.45(d,J＝7.2Hz,1H),6.55(d,J＝7.2Hz,1H),4.63–4.56(m,1H),4.05(s,3H),3.62–3.54(m,2H),3.32–3.28(m,1H),3.27–3.23(m,4H),3.20–2.99(m,4H),2.75–2.70(m,2H),2.39–2.19(m,4H),1.80–1.75(m,2H).LC-MS:ESI m/z 449.3[M+H] +；C 25H 32N 6O 2计算值448.26.HPLC纯度:100.0％(214nm),100.0％(254nm).
化合物27b： 1H NMR(400MHz,DMSO-d 6,80℃)δ7.96(s,1H),7.70(s,1H),7.55(d,J＝8.4Hz,1H),7.44(d,J＝8.4Hz,1H),7.02(d,J＝7.2Hz,1H),6.29(d,J＝7.2Hz,1H),4.11–4.08(m,1H),4.02(s,3H),3.29–3.23(m,2H),3.06–2.97(m,1H),2.77–2.68(m,3H),2.66–2.56(m,7H),2.44–2.42(m,2H),1.85–1.79(m,3H),1.48–1.38(m,1H).LC-MS:ESI m/z 449.3[M+H] +；C 25H 32N 6O 2计算值448.26.HPLC纯度:99.2％(214nm),99.7％(254nm).
制备得到化合物28，通过制备型HPLC(色谱柱：Kromasil-C18 100×21.2mm 5um；流动相：乙腈-水(0.1％甲酸)；梯度：10～20％)纯化粗品，得到呈黄色固体的化合物28a和化合物28b。化合物28a： 1H NMR(400MHz,DMSO-d 6,80℃)δ7.83(s,1H),7.75–7.69(m,2H),7.54(t,J＝7.6Hz,1H),7.08(d,J＝7.2Hz,1H),6.30(d,J＝7.2Hz,1H),6.22(brs,1H),4.11(t,J＝7.2Hz,1H),3.26–3.08(m,7H),2.87–2.79(m,1H),2.78(t,J＝7.2Hz,2H),2.73–2.58(m,4H),2.54(d,J＝6.4Hz,1H),2.05–2.03(m,1H),1.92–1.83(m,1H),1.81–1.74(m,2H).LC-MS:ESI m/z  420.2[M+H] +；C 24H 29N 5O 2计算值419.23.HPLC纯度:94.2％(214nm),93.3％(254nm).
化合物28b： 1H NMR(400MHz,DMSO-d 6,80℃)δ7.84(s,1H),7.74(d,J＝7.6Hz,2H),7.69(d,J＝7.6Hz,1H),7.54(t,J＝7.6Hz,1H),7.14(d,J＝7.2Hz,1H),6.46(brs,1H),6.37(d,J＝7.2Hz,1H),4.09(t,J＝7.2Hz,1H),3.42–3.21(m,7H),2.91(t,J＝7.2Hz,2H),2.75–2.61(m,4H),2.58–2.48(m,2H),2.06–1.90(m,1H),1.80–1.73(m,3H).LC-MS:ESI m/z 420.3[M+H] +；C 24H 29N 5O 2计算值419.23.HPLC纯度:99.4％(214nm),99.1％(254nm).
实施例11.化合物29的合成
Figure PCTCN2021078537-appb-000126
步骤1：1-(3-异丙氧基苯基)乙-1-酮
Figure PCTCN2021078537-appb-000127
将1-(3-羟苯基)乙酮(5.0g，37mmol)，2-溴丙烷(5.0g，40mmol)和K 2CO 3(7.6g，55mmol)在乙腈(60mL)中的混合物加热至70℃并搅拌16小时。浓缩混合物，倒入水(100mL)中，并用乙酸乙酯(120mL×3)萃取。合并的有机相用盐水(150mL)洗涤，经硫酸钠干燥，过滤并真空浓缩，得到残余物。残余物通过柱色谱法纯化(5～17％乙酸乙酯的石油醚溶液)，得到标题化合物(5.7g，83％)，为白色固体。 1H NMR(400MHz,DMSO-d 6)δ7.54–7.49(m,1H),7.45–7.39(m,2H),7.19–7.17(m,1H),4.72–4.66(m,1H),2.57(s,3H),1.28(d,J＝6.0Hz,6H).LC-MS:ESI m/z 179.2[M+H] +；C 11H 14O 2计算值178.10.
步骤2：3-(3-异丙氧基苯基)-3-氧代丙酸甲酯
Figure PCTCN2021078537-appb-000128
将1-(3-异丙氧基苯基)乙酮(2.0g，11mmol)和NaH(0.90g，22mmol)在碳酸二甲酯(15mL)中的混合物加热至50℃，搅拌3小时。将该混合物倒入NaHCO 3水溶液(50mL) 中，并用乙酸乙酯(60mL×3)萃取。合并的有机相用盐水(70mL×2)洗涤，经硫酸钠干燥，过滤并真空浓缩，得到黄色油状的标题化合物(2.1g，粗品)。 1H NMR(400MHz,DMSO-d 6)δ7.53–7.48(m,1H),7.47–7.40(m,2H),7.23–7.21(m,1H),4.72–4.66(m,1H),4.19(s,2H),3.65(s,3H),1.28(d,J＝6.0Hz,6H).LC-MS:ESI m/z 237.3[M+H] +；C 13H 16O 4计算值236.10.
步骤3-7：参照化合物23的合成步骤1-5进行，并且通过SFC(色谱柱：chiralpak-IG；流动相：CO 2-甲醇(含0.1％DEA))手性分离3-(3-异丙氧基苯基)-3-(((R)-1-(2-(5,6,7,8-四氢-1,8-萘啶-2-基)乙基)吡咯烷-3-基)氨基)丙酸(30mg，0.07mmol)，得到化合物29a和化合物29b。
化合物29a： 1H NMR(400MHz,DMSO-d 6)δ7.19(t,J＝8.0Hz,1H),7.00(d,J＝7.2Hz,1H),6.94(s,1H),6.89(d,J＝7.6Hz,1H),6.79(dd,J＝8.0,2.0Hz,1H),6.30–6.21(m,2H),4.63–4.57(m,1H),3.99–3.96(m,1H),3.31–3.19(m,2H),3.01–2.89(m,1H),2.68–2.52(m,7H),2.49–2.39(m,3H),2.38–2.25(m,2H),2.02–1.61(m,4H),1.26(d,J＝6.0Hz,6H).LC-MS:ESI m/z 453.3,455.3[M+H,Cl] +；C 26H 36N 4O 3计算值452.28.HPLC纯度:98.7％(214nm),96.2％(254nm).
化合物29b： 1H NMR(400MHz,DMSO-d 6)δ7.21(t,J＝8.0Hz,1H),7.03(d,J＝7.2Hz,1H),6.95(s,1H),6.90(d,J＝7.6Hz,1H),6.78(dd,J＝8.1,2.0Hz,1H),6.46(brs,1H),6.28(d,J＝7.3Hz,1H),4.63–4.57(m,1H),3.96–3.92(m,1H),3.26–3.20(m,2H),3.00–2.85(m,1H),2.71–2.52(m,9H),2.48–2.39(m,2H),2.35–2.26(m,1H),1.99–1.40(m,4H),1.26(d,J＝6.0Hz,6H).LC-MS:ESI m/z 453.3,455.3[M+H,Cl] +；C 26H 36N 4O 3计算值452.28.HPLC纯度:99.0％(214nm),99.3％(254nm).
实施例12.参照化合物29合成以下化合物：
化合物30(甲酸盐)： 1H NMR(400MHz,DMSO-d 6,80℃)δ7.34(d,J＝7.6Hz,1H),7.21(t,J＝7.2Hz,1H),7.06–6.95(m,2H),6.90(t,J＝7.2Hz,1H),6.30–6.23(m,1H),5.96(brs,1H),4.68–4.58(m,1H),4.40–4.28(m,1H),3.10–3.07(m,2H),2.80–2.66(m,5H),2.65–2.54(m,6H),2.43–2.35(m,2H),2.00–1.90(m,1H),1.79–1.71(m,2H),1.54–1.43(m,1H),1.31–1.29(m,6H).LC-MS:ESI m/z 453.3[M+H] +；C 26H 36N 4O 3计算值452.28.HPLC纯度:93.7％(214nm),92.3％(254nm).
化合物31(甲酸盐)： 1H NMR(400MHz,DMSO-d 6)δ7.29–7.26(m,2H),7.09–7.03(m,1H),6.86(d,J＝8.4Hz,2H),6.34–6.28(m,1H),4.65–4.53(m,1H),4.01–3.95(m,1H),3.28–3.25(m,2H),3.18–2.99(m,6H),2.80–2.53(m,6H),2.44-2.40(m,1H),2.05–1.52(m,4H),1.27(d,J＝6.0Hz,6H).LC-MS:ESI m/z 453.3[M+H] +；C 26H 36N 4O 3计算值452.28.HPLC纯度:96.8％(214nm),97.5％(254nm).
化合物32(甲酸盐)： 1H NMR(400MHz,DMSO-d 6)δ7.51–7.38(m,4H),7.08–7.00(m,1H),6.51(brs,1H),6.33–6.27(m,1H),4.05–3.98(m,1H),3.27–3.24(m,2H),2.91–2.74(m,6H),2.68–2.50(m,6H),2.44–2.41(m,1H),2.04–1.87(m,1H),1.81–1.50(m,3H).LC-MS:ESI m/z 429.3,430.2[M+H,Cl] +；C 23H 29ClN 4O 2计算值428.20.HPLC纯度:95.5％(214nm),94.7％(254nm).
化合物33(甲酸盐)： 1H NMR(400MHz,DMSO-d 6)δ7.54–7.37(m,3H),7.24(d,J＝8.0Hz,1H),7.08–7.02(m,1H),6.62(brs,1H),6.29–6.22(m,1H),4.07–4.03(m,1H),3.34–3.20(m,2H),2.98–2.94(m,1H),2.86–2.54(m,10H),2.46–2.34(m,2H),2.01–1.48(m,4H).LC-MS:ESI m/z 479.2[M+H] +；C 24H 29F 3N 4O 3计算值478.22.HPLC纯度:94.9％(214nm),95.2％(254 nm).
将化合物33通过SFC手性分离(色谱柱：Chiralpak-IG；流动相：CO 2-甲醇(0.1％DEA))得到化合物33a和化合物33b.
化合物33a： 1H NMR(400MHz,DMSO-d 6)δ7.49–7.36(m,3H),7.24(d,J＝8.0Hz,1H),7.00(d,J＝7.2Hz,1H),6.28(brs,1H),6.23(d,J＝7.2Hz,1H),4.07–4.03(m,1H),3.25–3.21(m,2H),2.92–2.85(m,1H),2.68–2.52(m,9H),2.44–2.31(m,2H),2.25–2.16(m,1H),2.03–1.60(m,4H)；LC-MS:ESI m/z 479.2[M+H] +；C 24H 29F 3N 4O 3计算值478.22.HPLC纯度:97.53％(214nm),97.05％(254nm).
化合物33b： 1H NMR(400MHz,DMSO-d 6)δ7.49–7.37(m,3H),7.23(d,J＝8.0Hz,1H),7.04(d,J＝7.2Hz,1H),6.60(brs,1H),6.28(d,J＝7.2Hz,1H),4.03–4.00(m,1H),3.25–3.21(m,2H),2.92–2.85(m,1H),2.67–2.52(m,9H),2.45–2.32(m,3H),2.02–1.37(m,4H).LC-MS:ESI m/z 479.2[M+H] +；C 24H 29F 3N 4O 3计算值478.22.HPLC纯度:95.6％(214nm),97.6％(254nm).
化合物34(甲酸盐)： 1H NMR(400MHz,DMSO-d 6,80℃)δ7.22(t,J＝7.7Hz,1H),6.98–6.91(m,3H),6.80(d,J＝8.0Hz,1H),6.25–6.24(m,1H),5.93(brs,1H),3.98(s,1H),3.75(s,3H),3.07–2.96(m,3H),2.62–2.54(m,8H),2.40–2.19(m,4H),1.87–1.60(m,4H).LC-MS:ESI m/z 425.3[M+H] +；C 24H 32N 4O 3计算值424.25.HPLC纯度:100.0％(214nm),99.8％(254nm).
化合物35(甲酸盐)： 1H NMR(400MHz,DMSO-d 6)δ7.24(t,J＝8.0Hz,1H),7.10–7.05(m,1H),6.98–6.90(m,2H),6.81(d,J＝8.0Hz,1H),6.40(brs,1H),6.33–6.28(m,1H),4.02–3.94(m,3H),3.27–3.23(m,2H),3.10–2.85(m,7H),2.75–2.54(m,5H),2.45–2.36(m,1H),2.11–1.80(m,1H),1.79–1.71(m,2H),1.62–1.58(m,1H),1.33(t,J＝7.6Hz,3H).LC-MS:ESI m/z 439.3[M+H] +；C 25H 34N 4O 3计算值438.26.HPLC纯度:98.2％(214nm),98.0％(254nm).
化合物36(甲酸盐)： 1H NMR(400MHz,DMSO-d 6,80℃)δ7.20–7.06(m,3H),7.04–6.91(m,2H),6.30–6.27(m,1H),3.97–3.91(m,1H),3.27–3.23(m,2H),3.04–2.96(m,1H),2.74–2.53(m,8H),2.42–2.23(m,4H),1.94–1.82(m,2H),1.79–1.74(m,2H),1.45–1.35(m,1H),0.96–0.88(m,2H),0.69–0.62(m,2H).LC-MS:ESI m/z 435.3[M+H] +；C 26H 34N 4O 2计算值434.27.HPLC纯度:95.7％(254nm),97.9％(214nm).
化合物37(甲酸盐)： 1H NMR(400MHz,DMSO-d 6)δ8.36(s,1H),7.64(s,1H),7.18(d,J＝7.6Hz,1H),7.01–6.96(m,1H),6.51(brs,1H),6.21(d,J＝7.2Hz,1H),3.94(s,1H),3.20–3.19(m,2H),2.85(s,1H),2.65–2.50(m,5H),2.45–2.10(m,10H),1.78–1.65(m,3H),1.56–1.33(m,1H).LC-MS:ESI m/z 410.2[M+H] +；C 23H 31N 5O 2计算值409.25.HPLC纯度:99.7％(214nm),99.8％(254nm).
化合物38(铵盐)： 1H NMR(400MHz,DMSO-d 6,80℃)δ8.08(s,1H),7.74–7.65(m,1H),7.00–6.97(m,1H),6.76(d,J＝8.4Hz,1H),6.29–6.23(m,1H),3.96–3.89(m,1H),3.80(s,3H),3.24–3.13(m,2H),2.89–2.87(m,1H),2.64–2.49(m,8H),2.47–2.29(m,3H),2.28–2.09(m,1H),1.91–1.77(m,1H),1.73–1.69(m,2H),1.62–1.30(m,1H).LC-MS:ESI m/z 426.3[M+H] +；C 23H 31N 5O 3计算值425.24.HPLC纯度:100.0％(214nm),100.0％(254nm).
化合物39(铵盐)： 1H NMR(400MHz,DMSO-d 6)δ7.56(s,1H),7.42(d,J＝7.6Hz,1H),7.41–7.35(m,1H),7.29–7.25(m,1H),7.03–6.98(m,1H),6.55(brs,1H),6.28–6.22(m,1H),3.99–3.88(m,1H),3.22–3.21(m,2H),2.98–2.80(m,1H),2.67–2.51(m,6H),2.44–2.12(m,6H),1.90–1.67(m,3H),1.56–1.34(m,1H).LC-MS:ESI m/z 473.1,475.1[M+H,Br] +；C 23H 29BrN 4O 2 计算值472.15.HPLC纯度:100.0％(214nm),99.6％(254nm).
将化合物39通过制备型HPLC进行纯化(色谱柱：Xtimate 10u C18 250×30；流动相：乙腈-水(0.1％甲酸)；梯度：10-20％，9分钟；流速：50mL/min)，得到化合物39a和化合物39b。
化合物39a： 1H NMR(400MHz,DMSO-d 6)δ7.60(d,J＝1.6Hz,1H),7.45(d,J＝7.6Hz,1H),7.38(d,J＝7.2Hz,1H),7.29(t,J＝7.6Hz,1H),7.05(d,J＝7.2Hz,1H),6.43(brs,1H),6.27(d,J＝7.2Hz,1H),4.00(t,J＝7.2Hz,1H),3.26–3.21(m,2H),3.03–2.78(m,6H),2.68–2.53(m,6H),2.44–2.38(m,1H),1.98–1.89(m,1H),1.75–1.74(m,3H).LC-MS:ESI m/z 473.2[M+H] +；C 23H 29BrN 4O 2计算值472.15.HPLC纯度:98.2％(214nm),97.9％(254nm).
化合物39b： 1H NMR(400MHz,DMSO-d 6)δ7.60(d,J＝1.6Hz,1H),7.45(d,J＝7.6Hz,1H),7.39(d,J＝7.6Hz,3H),7.29(t,J＝7.6Hz,1H),7.08(d,J＝7.2Hz,1H),6.63(brs,1H),6.31(d,J＝7.2Hz,1H),3.97(t,J＝7.2Hz,1H),3.27–3.22(m,2H),3.13–2.76(m,7H),2.75–2.68(m,2H),2.61(t,J＝6.0Hz,2H),2.57–2.52(m,1H),2.44–2.38(m,1H),1.98–1.88(m,1H),1.79–1.71(m,2H),1.55–1.49(m,1H).LC-MS:ESI m/z 473.2[M+H] +,C 23H 29BrN 4O 2计算值472.15.HPLC纯度:99.2％(214nm),99.0％(254nm).
化合物40(甲酸盐)： 1H NMR(400MHz,DMSO-d 6)δ7.12(d,J＝7.2Hz,1H),7.00(t,J＝7.2Hz,1H),6.82–6.76(m,2H),6.45(brs,1H),6.27–6.23(m,1H),4.49(t,J＝8.4Hz,2H),3.91–3.82(m,1H),3.24–3.20(m,2H),3.12(t,J＝8.4Hz,2H),2.99–2.79(m,1H),2.60–2.53(m,5H),2.47–2.27(m,5H),2.22–2.10(m,2H),1.87–1.77(m,1H),1.76–1.71(m,2H),1.67–1.31(m,1H).LC-MS:ESI m/z 437.3[M+H] +；C 25H 32N 4O 3计算值436.25.HPLC纯度:99.0％(214nm),98.4％(254nm).
化合物42(甲酸盐)： 1H NMR(400MHz,DMSO-d 6,80℃)δ7.90–7.82(m,4H),7.60–7.54(m,1H),7.54–7.46(m,2H),7.04–6.97(m,1H),6.29–6.21(m,1H),5.93(brs,1H),4.23–4.10(m,1H),3.03–2.98(m,2H),2.97–2.88(m,1H),2.70–2.55(m,7H),2.48–2.37(m,4H),2.25–2.22(m,1H),2.07–1.85(m,1H),1.78–1.76(m,2H),1.70–1.45(m,1H).LC-MS:ESI m/z 445.3[M+H] +；C 27H 32N 4O 2计算值445.25.HPLC纯度:92.7％(214nm),98.0％(254nm).
化合物43(甲酸盐)： 1H NMR(400MHz,DMSO-d 6)δ7.26–7.22(m,1H),7.00–6.95(m,3H),6.80(brs,1H),6.29–6.23(m,1H),4.00–3.95(m,1H),3.79–3.78(m,2H),3.23–3.21(m,3H),3.01(s,1H),2.80–2.54(m,7H),2.37–2.32(m,2H),2.06–1.86(m,2H),1.73–1.70(m,2H),1.52–1.22(m,2H),0.86–0.63(m,4H).LC-MS:ESI m/z 451.3[M+H] +；C 26H 34N 4O 3计算值450.26.HPLC纯度:95.5％(214nm),100.0％(254nm).
化合物45(甲酸盐)： 1H NMR(400MHz,DMSO-d 6,80℃)δ7.22(t,J＝8.0Hz,1H),7.06–6.90(m,3H),6.82(d,J＝8.0Hz,1H),6.30–6.25(m,1H),5.97(brs,1H),4.12–4.09(m,2H),3.97(s,1H),3.70–3.64(m,2H),3.24(s,3H),3.22–3.20(m,2H),3.02(s,1H),2.76–2.51(m,9H),2.45–2.19(m,3H),1.88–1.81(m,1H),1.79–1.72(m,2H),1.63–1.49(m,1H).LC-MS:ESI m/z 469.3[M+H] +；C 26H 36N 4O 4计算值468.2.HPLC纯度:95.5％(214nm),95.3％(254nm).
制备得到化合物46，粗产物通过制备型HPLC(柱：Xbridge 5u C18 150×19mm；流动相：乙腈-水(0.05％甲酸)；梯度：5～15％乙腈，流速：20mL/min)纯化，得到化合物46a和化合物46b.
化合物46a： 1H NMR(400MHz,DMSO-d 6,80℃)δ7.70(s,1H),7.66(d,J＝7.2Hz,1H),7.61–7.50(m,2H),7.00(d,J＝7.2Hz,1H),6.24(d,J＝7.2Hz,1H),5.80(brs,1H),4.12(t,J＝6.8Hz,1H),3.30–3.16(m,2H),3.08–3.00(m,1H),2.85–2.63(m,5H),2.61–2.59(m,5H),2.46–2.32(m,2H),1.96–1.87(m,1H),1.79–1.71(m,2H),1.70–1.56(m,1H).LC-MS:ESI m/z 463.3[M+H] +；C 24H 29F 3N 4O 2计算值462.22.HPLC纯度:93.9％(214nm),95.9％(254nm).
化合物46b： 1H NMR(400MHz,DMSO-d 6,80℃)δ7.71(s,1H),7.67(d,J＝7.2Hz,1H),7.59–7.51(m,2H),7.03(d,J＝7.2Hz,1H),6.29(d,J＝7.2Hz,1H),4.13–4.05(m,1H),3.30–3.21(m,2H),2.97–2.95(m,1H),2.82–2.66(m,3H),2.66–2.50(m,8H),2.46–2.42(m,1H),1.90–1.80(m,1H),1.80–1.71(m,2H),1.48–1.36(m,1H).LC-MS:ESI m/z 463.3[M+H] +；C 24H 29F 3N 4O 2计算值462.22.HPLC纯度:96.5％(214nm),96.6％(254nm).
制备得到化合物49，通过制备型HPLC(色谱柱：Kromasil-C18 100×21.2mm 5um；流动相：乙腈-水(0.1％甲酸)；梯度：10～20％乙腈)纯化，得到化合物49a和化合物49b。
化合物49a： 1H NMR(400MHz,DMSO-d 6)δ7.60(d,J＝7.1Hz,1H),7.39(d,J＝7.9Hz,1H),7.34(t,J＝7.1Hz,1H),7.28–7.22(m,1H),7.00(d,J＝7.3Hz,1H),6.28–6.23(m,2H),4.46(t,J＝6.7Hz,1H),3.22–3.19(m,2H),2.91–2.89(m,1H),2.69–2.51(m,9H),2.39–2.38(m,2H),2.29–2.28(m,1H),1.87–1.82(m,1H),1.75–1.61(m,3H)；LC-MS:ESI m/z 429.2,431.2[M+H,Cl] +；C 23H 29ClN 4O 2计算值428.20.HPLC纯度:98.9％(214nm),99.1％(254nm).
化合物49b： 1H NMR(400MHz,DMSO-d 6)δ7.61(d,J＝7.7Hz,1H),7.40–7.30(m,2H),7.26–7.24(m,1H),7.04(d,J＝7.3Hz,1H),6.71(brs,1H),6.28(d,J＝7.2Hz,1H),4.44–4.39(m,1H),3.23–3.20(m,2H),2.88–2.86(m,1H),2.73–2.55(m,9H),2.45–2.30(m,3H),1.96–1.84(m,1H),1.75–1.69(m,2H),1.42–1.39(m,1H).LC-MS:ESI m/z 429.2,431.2[M+H,Cl] +；C 23H 29ClN 4O 2计算值428.20.HPLC纯度:97.9％(214nm),96.3％(254nm).
制备得到化合物51，将其通过制备型HPLC纯化(色谱柱：Xbridge 5u C18 150×19mm；流动相：乙腈-水(0.05％甲酸)；梯度：5～15％乙腈，流速：20mL/min)得到化合物51a和化合物51b。
化合物51a： 1H NMR(400MHz,DMSO-d 6,80℃)δ7.21(t,J＝8.0Hz,1H),7.02(d,J＝7.2Hz,1H),6.96–6.88(m,2H),6.78(dd,J＝8.0,2.4Hz,1H),6.27(d,J＝7.2Hz,1H),4.61–4.55(m,1H),3.99(dd,J＝8.8,5.6Hz,1H),3.30–3.22(m,2H),3.11–3.08(m,1H),2.85–2.65(m,5H),2.64–2.52(m,5H),2.49–2.36(m,2H),1.98–1.88(m,1H),1.84–1.73(m,2H),1.72–1.64(m,1H),1.27(d,J＝6.0Hz,6H).LC-MS:ESI m/z 453.3[M+H] +；C 26H 36N 4O 3计算值452.28.HPLC纯度:94.9％(214nm),94.4％(254nm).
化合物51b： 1H NMR(400MHz,DMSO-d 6,80℃)δ7.21(t,J＝8.0Hz,1H),7.04(d,J＝7.2Hz,1H),6.96–6.86(m,2H),6.78(dd,J＝8.0,2.4Hz,1H),6.30(d,J＝7.2Hz,1H),4.61–4.55(m,1H),3.95(dd,J＝8.4,5.6Hz,1H),3.32–3.22(m,2H),3.11–3.08(m,1H),2.88–2.52(m,11H),2.42–2.38(m,1H),1.90–1.83(m,1H),1.82–1.73(m,2H),1.52–1.44(m,1H),1.27(d,J＝6.0Hz,6H).LC-MS:ESI m/z 453.3[M+H] +；C 26H 36N 4O 3计算值452.28.HPLC纯度:95.0％(214nm),94.1％(254nm).
制备得到化合物55，通过制备型HPLC(色谱柱：Kromasil-C18 100×21.2mm 5um；流动相： 乙腈-水(0.1％甲酸)；梯度：2～8％乙腈)纯化，得到化合物55a和化合物55b。
化合物55a： 1H NMR(400MHz,DMSO-d 6)δ8.53(d,J＝1.6Hz,1H),8.46(dd,J＝4.8,1.6Hz,1H),7.79(dt,J＝8.0,2.0Hz,1H),7.35(dd,J＝7.6,4.8Hz,1H),7.00(d,J＝7.2Hz,1H),6.29(brs,1H),6.24(d,J＝7.2Hz,1H),4.03–4.00(m,1H),3.24–3.19(m,2H),2.94–2.86(m,1H),2.63–2.52(m,8H),2.48–2.36(m,3H),2.20–2.17(m,1H),1.90–1.82(m,1H),1.76–1.69(m,2H),1.64–1.55(m,1H)；LC-MS:ESI m/z 396.3[M+H] +；C 22H 29N 5O 2计算值395.23.HPLC纯度:99.9％(214nm),99.9％(254nm).
化合物55b： 1H NMR(400MHz,DMSO-d 6)δ8.56(d,J＝1.6Hz,1H),8.46(dd,J＝4.8,1.6Hz,1H),8.23(s,2H),7.81(d,J＝8.0Hz,1H),7.36(dd,J＝7.6,4.8Hz,1H),7.07(d,J＝7.2Hz,1H),6.62(brs,1H),6.30(d,J＝7.2Hz,1H),4.00–3.96(m,1H),3.27–3.21(m,2H),2.97–2.59(m,11H),2.48–2.36(m,2H),1.90–1.82(m,1H),1.78–1.71(m,2H),1.50–1.13(m,1H).LC-MS:ESI m/z 396.3[M+H] +；C 22H 29N 5O 2计算值395.51.HPLC纯度:94.3％(214nm),96.2％(254nm).
化合物57(铵盐)： 1H NMR(400MHz,DMSO-d 6,80℃)δ8.29(s,1H),8.22(s,1H),7.51(s,1H),6.98–6.95(m,1H),6.27–6.19(m,1H),5.93(brs,1H),3.97–3.88(m,1H),3.26–3.20(m,2H),2.98–2.84(m,1H),2.70–2.48(m,8H),2.42–2.07(m,7H),1.92–1.69(m,3H),1.65–1.26(m,1H).LC-MS:ESI m/z 410.3[M+H] +；C 23H 31N 5O 2计算值409.25.HPLC纯度:99.8％(214nm),100.0％(254nm).
化合物58(甲酸盐)： 1H NMR(400MHz,DMSO-d 6)δ8.25(brs,1H),8.22–8.10(m,2H),7.43(d,J＝5.6Hz,1H),7.04(t,J＝8.0Hz,1H),6.36(brs,1H),6.30–6.22(m,1H),4.08–4.02(m,1H),3.89–3.77(m,3H),3.28–3.20(m,2H),3.02–2.96(m,1H),2.88–2.65(m,5H),2.64–2.54(m,5H),2.46–2.37(m,2H),2.00–1.88(m,1H),1.80–1.63(m,3H).LC-MS:ESI m/z 426.3[M+H] +；C 23H 31N 5O 3计算值425.24.HPLC纯度:98.8％(214nm),100.0％(254nm).
实施例13.化合物63的合成
Figure PCTCN2021078537-appb-000129
步骤1：1-(3-溴苯基)-3-甲基-1H-吡唑(a)和1-(3-溴苯基)-5-甲基-1H-吡唑(b)
Figure PCTCN2021078537-appb-000130
将(3-溴苯基)肼盐酸盐(3.0g，13mmol)，3-丁炔-2-酮(0.91g，13mmol)，浓盐酸(0.20mL)溶于甲醇(12mL)中，在120℃下微波反应30分钟。将反应混合物浓缩至干，并用二氯甲烷(10mL)和水(10mL)稀释。将水相进一步用二氯甲烷(5mL×2)萃取。浓缩合并的有机相，并通过FCC纯化(5～10％乙酸乙酯的石油醚溶液)，得到1-(3-溴苯基)-3-甲基-1H-吡唑(1.2g，36％)和1-(3(-溴苯基)-5-甲基-1H-吡唑(3.2g，64％)，为黄色油状物。1-(3-溴苯基)-3-甲基-1H-吡唑(a): 1H NMR(400MHz,CDCl 3)δ7.93–7.85(m,1H),7.79(d,J＝2.4Hz,1H),7.68–7.51(m,1H),7.42–7.34(m,1H),7.34–7.24(m,1H),6.26(d,J＝2.4Hz,1H),2.37(s,3H).
1-(3-溴苯基)-5-甲基-1H-吡唑(b): 1H NMR(400MHz,CDCl 3)δ7.66(t,J＝2.0Hz,1H),7.58(d,J＝1.6Hz,1H),7.53–7.47(m,1H),7.43–7.38(m,1H),7.33(t,J＝8.0Hz,1H),6.20(d,J＝0.8Hz,1H),2.37(s,3H).LC-MS:ESI m/z 237.1,239.1[M+H,Br] +；C 10H 9BrN 2计算值235.99.
步骤2：1-(3-(5-甲基-1H-吡唑-1-基)苯基)乙-1-酮
Figure PCTCN2021078537-appb-000131
将1-(3-溴苯基)-5-甲基吡唑(2.0g，8.4mmol)，1-(乙烯氧基)丁烷(3.4g，34mmol)，二乙酸钯(0.19g，0.80mmol)和三乙胺(2.5g，25mmol)在1，4-二氧六环(20mL)中的混合物用N 2脱气3次。将反应混合物在N 2气氛下于115℃回流反应24小时。将混合物倒入盐酸(50mL，1N)中，并在室温搅拌1小时。用乙酸乙酯(30mL×3)萃取混合物。合并的有机相经硫酸钠干燥，过滤并浓缩至干。粗产物通过硅胶柱色谱法(10～25％乙酸乙酯的石油醚溶液)纯化，得到1-(3-(5-甲基-1H-吡唑-1-基)苯基)乙-1-酮(1.0g，51％)，为黄色油状物。 1H NMR(400MHz,DMSO-d 6)δ8.01(t,J＝2.0Hz,1H),7.98–7.95(m,1H),7.79(ddd,J＝8.0,2.0,1.2Hz,1H),7.65(t,J＝8.0Hz,1H),7.58(d,J＝1.6Hz,1H),6.30(s,1H),2.61(s,3H),2.35(s,3H).LC-MS:ESI m/z 201.3[M+H] +；C 12H 12N 2O计算值200.09.
步骤3：3-(3-(5-甲基-1H-吡唑-1-基)苯基)-3-氧代丙酸甲酯
Figure PCTCN2021078537-appb-000132
将氢化钠(0.80g，20mmol)和1-(3-(5-甲基-1H-吡唑-1-基)苯基)乙-1-酮(1.1g，10mmol)在碳酸二乙酯(20mL)的反应液于60℃下搅拌反应2小时。将混合物用饱和NH 4Cl(100mL) 淬灭，并用二氯甲烷(30mL×2)萃取。合并的有机相用盐水(50mL)洗涤，经硫酸钠干燥并浓缩至干。粗产物通过硅胶柱色谱法纯化(10～25％乙酸乙酯的石油醚溶液)。得到标题化合物(0.90g，33％)，为黄色油状物。 1H NMR(400MHz,DMSO-d 6)δ8.03(s,1H),7.97(d,J＝8.0Hz,1H),7.84(d,J＝8.0Hz,1H),7.68(t,J＝8.0Hz,1H),7.60(d,J＝1.2Hz,1H),6.31(s,1H),4.27(s,2H),3.64(s,3H),2.36(s,3H).LC-MS:ESI m/z 259.1[M+H] +；C 14H 14N 2O 3计算值258.10.
步骤4-8：参照化合物23的步骤1-5进行合成，得到化合物63(甲酸盐)：
1HNMR(400MHz,DMSO-d 6)δ7.62–7.27(m,5H),7.10–6.89(m,1H),6.26–6.20(m,2H),4.08–4.01(m,1H),3.20–3.19(m,2H),3.02–2.88(m,1H),2.75–2.50(m,10H),2.44–2.21(m,5H),1.93–1.41(m,4H).LC-MS:ESI m/z 475.3[M+H] +；C 27H 34N 6O 2计算值474.27.HPLC纯度:97.7％(214nm),97.3％(254nm).
实施例14.参照化合物63的合成方法，制备得到以下化合物：
化合物65(甲酸盐)： 1H NMR(400MHz,DMSO-d 6)δ7.81(s,1H),7.66(d,J＝8.0Hz,1H),7.41(t,J＝8.0Hz,1H),7.31–7.23(m,1H),7.02–6.97(m,1H),6.53(brs,1H),6.34–6.21(m,2H),4.08–4.01(m,1H),3.25–3.21(m,2H),3.01–2.93(m,1H),2.68–2.52(m,8H),2.46–2.36(m,3H),2.27(s,3H),2.25–2.20(m,1H),1.89–1.79(m,1H),1.78–1.63(m,2H),1.62–1.39(m,1H).LC-MS:ESI m/z 475.3[M+H] +；C 27H 34N 6O 2计算值474.27.HPLC纯度:99.1％(254nm),99.8％(214nm).
制备得到化合物65，通过SFC(色谱柱：ChiralPak-AD-H，0.46cm×25cm，流动相：正己烷/乙醇/二乙胺＝70/30/0.1(V/V))进行纯化，得到化合物65a和化合物65b。
化合物65a： 1H NMR(400MHz,DMSO-d 6,80℃)δ7.77(s,1H),7.62(d,J＝8.0Hz,1H),7.39(t,J＝8.0Hz,1H),7.26(d,J＝7.6Hz,1H),7.01(d,J＝7.2Hz,1H),6.30–6.27(m,2H),6.13(brs,1H),4.06–4.03(m,1H),3.28–3.22(m,2H),3.07–2.96(m,1H),2.70–2.52(m,9H),2.48–2.38(m,3H),2.28(s,3H),1.85–1.74(m,3H),1.43–1.38(m,1H).LC-MS:ESI m/z 475.3[M+H] +；C 27H 34N 6O 2计算值474.27.HPLC纯度:96.9％(214nm),96.7％(254nm).
化合物65b： 1H NMR(400MHz,DMSO-d 6,80℃)δ7.76(s,1H),7.62(d,J＝8.0Hz,1H),7.39(t,J＝8.0Hz,1H),7.25(d,J＝7.6Hz,1H),6.98(d,J＝7.2Hz,1H),6.29(d,J＝2.2Hz,1H),6.23(d,J＝7.2Hz,1H),5.94(brs,1H),4.09–4.06(m,1H),3.23–3.22(m,2H),3.07–3.03(m,1H),2.72–2.50(m,10H),2.44–2.42(m,1H),2.28–2.27(m,4H),1.91–1.88(m,1H),1.75–1.72(m,2H),1.62–1.60(m,1H).LC-MS:ESI m/z 475.3[M+H] +；C 27H 34N 6O 2计算值474.27.HPLC纯度:99.1％(214nm),98.5％(254nm).
化合物68(甲酸盐)： 1H NMR(400MHz,DMSO-d 6,80℃)δ7.54–7.52(m,1H),7.50–7.43(m,2H),7.41–7.37(m,1H),7.02–6.98(m,1H),6.29–6.23(m,1H),5.95(brs,1H),4.10–4.07(m,1H),3.26–3.23(m,2H),3.03–3.02(m,1H),2.73–2.52(m,10H),2.48–2.44(m,2H),2.40(s,3H),2.27(s,3H),1.94–1.80(m,1H),1.79–1.71(m,2H),1.64–1.43(m,1H).LC-MS:ESI m/z 490.3[M+H] +；C 27H 35N 7O 2计算值489.29.HPLC纯度:99.6％(214nm),100.0％(254nm).
制备得到化合物70，将混合物通过制备型HPLC纯化(色谱柱：Kromasil C18 5um 100×21.5mm；流动相：乙腈-水(0.1％甲酸)；梯度：89-99％乙腈，6.5分钟；流速：25mL/min)，得 到化合物70a和化合物70b。
化合物70a： 1H NMR(400MHz,DMSO-d 6)δ8.29(s,1H),7.63(s,1H),7.56(d,J＝8.8Hz,1H),7.29(d,J＝8.8Hz,1H),7.02(d,J＝7.2Hz,1H),6.35(brs,1H),6.24(d,J＝7.2Hz,1H),4.15(s,3H),4.12–4.08(m,1H),3.24–3.20(m,2H),3.09–3.08(m,1H),2.87–2.74(m,4H),2.70–2.65(m,2H),2.64–2.50(m,5H),2.43–2.39(m,1H),1.97–1.91(m,1H),1.76–1.71(m,3H).LC-MS:ESI m/z 449.3[M+H] +；C 25H 32N 6O 2计算值448.26.HPLC纯度:99.2％(214nm),100.0％(254nm).
化合物70b： 1H NMR(400MHz,DMSO-d 6)δ8.29(s,1H),7.63(s,1H),7.57(d,J＝8.8Hz,1H),7.30(d,J＝8.8Hz,1H),7.09(d,J＝7.2Hz,1H),6.59(brs,1H),6.32(d,J＝7.2Hz,1H),4.15(s,3H),4.10–4.05(m,1H),3.27–3.22(m,2H),3.12–2.97(m,6H),2.93–2.79(m,1H),2.73(t,J＝7.2Hz,2H),2.67–2.60(m,3H),2.48–2.42(m,1H),1.92–1.87(m,1H),1.77–1.72(m,2H),1.71–1.51(m,1H).LC-MS:ESI m/z 449.3[M+H] +；C 25H 32N 6O 2计算值448.26.HPLC纯度:97.8％(214nm),97.7％(254nm).
实施例15.化合物72的合成
Figure PCTCN2021078537-appb-000133
步骤1：3-(3-吗啉代苯基)-3-氧代丙酸甲酯
Figure PCTCN2021078537-appb-000134
向3-吗啉代苯甲酸(3.8g，18mmol)的无水四氢呋喃(40mL)溶液中加入NaCl(52mg，0.90mmol)和亚硫酰氯(3.9mL，66mmol)。将混合物在N 2气氛下于75℃下搅拌2小时。将混合物浓缩至干。在25℃下，向丙二酸乙酯钾盐(3.0g，25mmol)的无水四氢呋喃(40mL)溶液中加入MgCl 2(5.6g，58mmol)。将混合物在25℃下搅拌0.5小时。在25℃下将三乙胺(7.5mL，54mmol)加入溶液中，然后搅拌1小时。将以上制备的3-吗啉代苯甲酰氯在无水四氢呋喃(40mL)中的混合物加入该溶液中。该混合物在80℃下搅拌2.5小时。将混合物浓缩至干。将残留物用水(100mL)稀释，并用乙酸乙酯(100mL×2)萃取。合并有机相并用盐水(100mL)洗涤，经硫酸钠干燥并浓缩至干。残余物通过快速色谱法纯化(石油醚：乙 酸乙酯＝1：1)，得到标题化合物(3.5g，66％)，为黄色油状物。 1H NMR(400MHz,DMSO-d 6)δ7.45–7.41(m,1H),7.41–7.34(m,2H),7.29–7.23(m,1H),4.20(s,2H),3.78–3.74(m,4H),3.65(s,3H),3.19–3.15(m,4H).LC-MS:ESI m/z 264.2[M+H] +；C 14H 17NO 4计算值263.12.
步骤2-6：参照前述化合物如化合物63等的合成方法由3-(3-吗啉代苯基)-3-氧代丙酸甲酯进一步制备得到化合物72(甲酸盐)： 1H NMR(400MHz,DMSO-d 6)δ7.61–7.54(m,1H),7.46–7.42(m,1H),7.27–7.24(m,1H),7.07–7.03(m,1H),6.95(d,J＝8.4Hz,1H),6.71–6.66(m,1H),4.54–4.48(m,1H),3.75–3.71(m,4H),3.71–3.56(m,6H),3.42–3.40(m,2H),3.35–3.24(m,2H),3.22–3.17(m,5H),3.15–3.02(m,2H),2.76–2.70(m,2H),2.37–2.23(m,2H),1.87–1.78(m,2H).LC-MS:ESI m/z 480.3[M+H] +；C 27H 37N 5O 3计算值479.29.HPLC纯度:95.5％(254nm),95.5％(214nm).
实施例16.参照化合物72的合成方法，制备以下化合物：
化合物73(甲酸盐)： 1H NMR(400MHz,DMSO-d 6)δ7.41(s,1H),7.27–7.11(m,3H),7.08–7.00(m,1H),6.34–6.27(m,1H),4.06–4.03(m,1H),3.28–3.24(m,2H),3.14(s,1H),3.05–2.76(m,6H),2.75–2.56(m,5H),2.47–2.43(m,1H),2.02–1.85(m,1H),1.77–1.75(m,2H),1.58–1.55(m,1H),1.28(s,9H).LC-MS:ESI m/z 451.3[M+H] +；C 27H 38N 4O 2计算值450.30.HPLC纯度:94.5％(214nm),94.7％(254nm).
化合物74(铵盐)： 1H NMR(400MHz,DMSO-d 6)δ8.07–8.06(m,1H),7.71–7.64(m,1H),7.04–6.95(m,1H),6.71(d,J＝8.4Hz,1H),6.49(s,1H),6.29–6.23(m,1H),4.29–4.24(m,2H),3.96–3.89(m,1H),3.24–3.21(m,2H),2.90–2.88(m,1H),2.67–2.54(m,7H),2.48–2.28(m,4H),1.89–1.73(m,3H),1.63–1.34(m,1H),1.33–1.26(m,3H).LC-MS:ESI m/z 440.3[M+H] +；C 24H 33N 5O 3计算值439.26.HPLC纯度:100.0％(214nm),99.8％(254nm).
化合物77(铵盐)： 1H NMR(400MHz,DMSO-d 6)δ8.68–8.67(m,1H),8.00–7.97(m,1H),7.73–7.71(m,1H),7.00–6.96(m,1H),6.27–6.22(m,1H),5.94(brs,1H),4.05–4.01(m,1H),3.20–3.18(m,2H),2.93–2.84(m,1H),2.67–2.51(m,7H),2.44–2.05(m,5H),1.89–1.71(m,3H),1.65–1.29(m,1H).LC-MS:ESI m/z 464.3[M+H] +；C 23H 28F 3N 5O 2计算值463.22.HPLC纯度:100.0％(254nm),100.0％(214nm).
化合物78(甲酸盐)： 1H NMR(400MHz,DMSO-d 6)δ8.08(s,1H),7.72–7.70(m,1H),7.07–7.04(m,1H),6.72(d,J＝8.0Hz,1H),6.60–6.41(m,1H),6.31–6.27(m,1H),5.25–5.19(m,1H),4.00–3.92(m,1H),3.24–3.22(m,2H),3.05–2.56(m,11H),2.48–2.29(m,1H),1.96–1.85(m,1H),1.77–1.70(m,2H),1.53–1.50(m,1H),1.27(d,J＝6.0Hz,6H).LC-MS:ESI m/z 454.3[M+H] +；C 25H 35N 5O 3计算值453.27.HPLC纯度:99.2％(214nm),98.9％(254nm).
化合物81(铵盐)： 1H NMR(400MHz,DMSO-d 6)δ9.07–9.06(m,1H),8.78–8.77(m,2H),7.05–6.98(m,1H),6.69(brs,1H),6.28–6.22(m,1H),4.01–3.95(m,1H),3.22–3.19(m,2H),2.93–2.85(m,1H),2.73–2.51(m,11H),2.18–2.16(m,1H),1.95–1.70(m,3H),1.60–1.35(m,1H).LC-MS:ESI m/z 397.3[M+H] +；C 21H 28N 6O 2计算值396.23.HPLC纯度:99.5％(214nm),99.6％(254nm).
化合物82(铵盐)： 1H NMR(400MHz,DMSO-d 6,80℃)δ8.59–8.58(m,2H),6.99(t,J＝7.2Hz,1H),6.28–6.23(m,1H),5.94(brs,1H),3.95–3.90(m,1H),3.26–3.23(m,2H),2.93–2.92(m,1H),2.67–2.58(m,4H),2.57(s,3H),2.55–2.50(m,2H),2.49–2.30(m,5H),2.30–2.08(m,1H),1.94–1.78(m,1H),1.80–1.74(m,2H),1.58–1.27(m,1H).LC-MS:ESI m/z 411.3[M+H] +；C 22H 30N 6O 2计算值410.24.HPLC纯度:100.0％(254nm),100.0％(214nm).
化合物86(铵盐)： 1H NMR(400MHz,DMSO-d 6,80℃)δ7.54(s,1H),7.39(d,J＝7.6Hz,1H),7.27(t,J＝7.6Hz,1H),7.15(d,J＝7.2Hz,1H),7.03–6.94(m,1H),6.28–6.23(m,1H),5.94(brs,1H),4.02–3.95(m,1H),3.26–3.23(m,2H),3.03–2.98(m,1H),2.70–2.54(m,6H),2.48–2.34(m,4H),2.31(s,3H),2.29–2.19(s,5H),1.80–1.74(m,3H),1.62–1.39(m,1H).LC-MS:ESI m/z 489.3[M+H] +；C 28H 36N 6O 2计算值488.29.HPLC纯度:98.8％(214nm),99.5％(254nm).
制备得到化合物91，通过制备型HPLC纯化(色谱柱：Kromasil-C18 100×21.2mm 5um；流动相：乙腈-水(0.1％三氟乙酸)；梯度：25～35％)，得到化合物91a和化合物91b。
化合物91a： 1H NMR(400MHz,DMSO-d 6,80℃)δ7.69(s,1H),7.52–7.47(m,2H),6.98(d,J＝7.2Hz,1H),6.22(d,J＝7.2Hz,1H),6.05(brs,1H),4.15–4.06(m,1H),3.25–3.20(m,2H),3.00–2.95(m,1H),2.72–2.62(m,3H),2.59–2.54(m,8H),2.32–2.26(m,1H),1.95–1.84(m,1H),1.77–1.70(m,2H),1.69–1.56(m,1H),1.30(s,9H).LC-MS:ESI m/z 519.3[M+H] +；C 28H 37F 3N 4O 2计算值518.29.HPLC纯度:99.8％(214nm),99.4％(254nm).
化合物91b： 1H NMR(400MHz,DMSO-d 6)δ7.70(s,1H),7.52–7.47(m,2H),7.01(d,J＝7.2Hz,1H),6.27(d,J＝7.2Hz,1H),3.93–3.90(m,1H),3.28–3.21(m,2H),3.0–2.92(m,1H),2.72–2.64(m,3H),2.62–2.52(m,7H),2.46–2.38(m,2H),1.84–1.71(m,3H),1.47–1.39(m,1H),1.31(s,9H).LC-MS:ESI m/z 519.3[M+H] +；C 28H 37F 3N 4O 2计算值518.29.HPLC纯度:97.6％(214nm),98.7％(254nm).
化合物95(铵盐)： 1H NMR(400MHz,DMSO-d 6,80℃)δ7.98(s,1H),7.68(s,1H),7.47(d,J＝8.4Hz,1H),7.36(d,J＝8.4Hz,1H),6.98–6.95(m,1H),6.27–6.20(m,1H),5.91(brs,1H),4.19–3.99(m,1H),3.25–3.23(m,2H),3.05–2.96(m,1H),2.70–2.50(m,8H),2.45–2.17(m,4H),1.94–1.77(m,1H),1.77–1.72(m,2H),1.65–1.36(m,1H).LC-MS:ESI m/z 435.2[M+H] +；C 24H 30N 6O 2计算值434.24.HPLC纯度:99.6％(254nm),99.3％(214nm).
化合物96(铵盐)： 1H NMR(400MHz,DMSO-d 6,80℃)δ8.85–8.86(m,2H),8.13(s,1H),7.05–6.94(m,1H),6.29–6.18(m,1H),5.92(brs,1H),4.16–4.14(m,1H),3.25–3.21(m,2H),2.94–2.93(m,1H),2.70–2.50(m,10H),2.44–2.12(m,2H),1.89–1.72(m,3H),1.58–1.33(m,1H).LC-MS:ESI m/z 464.3[M+H] +；C 23H 28F 3N 5O 2计算值463.22.HPLC纯度:100.0％(214nm),100.0％(254nm).
实施例17.化合物97的合成
Figure PCTCN2021078537-appb-000135
步骤1：3-(3,5-二甲基-1H-吡唑-1-基)苯甲酸
Figure PCTCN2021078537-appb-000136
在N 2气氛下，将3-肼基苯甲酸盐酸盐(5.0g，26.5mmol)加入到乙酸(50mL)和水(50mL)的混合溶剂中，加热直到固体完全溶解，然后逐滴加入2,4-戊二酮(2.6g，26.5mmol)。在70℃下搅拌3小时后，真空除去溶剂，并将冰水倒入反应液中。待白色固体沉淀析出，过滤收集固体，真空干燥，得到标题化合物，为浅黄色固体(4.1g，72％)。 1H NMR(400MHz,DMSO-d 6)δ13.23(s,1H),8.01(s,1H),7.93(d,J＝7.8Hz,1H),7.76(d,J＝1.1Hz,1H),7.63(d,J＝7.9Hz,1H),6.11(s,1H),2.33(s,3H),2.20(s,3H).LC-MS:ESI m/z 217.14[M+H] +；C 12H 12N 2O 2计算值216.09.
步骤2：(3-(3,5-二甲基-1H-吡唑-1-基)苯基)甲醇
Figure PCTCN2021078537-appb-000137
在N 2气氛下，向3-(3,5-二甲基-1H-吡唑-1-基)苯甲酸(2.0g，9.25mmol)的四氢呋喃(20mL)溶液中添加BH 3.THF(1N，34.6mL)。在75℃下搅拌16小时后，将反应混合物用1N盐酸水溶液淬灭，并用饱和NaHCO 3水溶液中和至pH≈8。反应混合物用二氯甲烷(20mL×3)萃取，合并的有机相依次用水(30mL)、盐水(30mL)洗涤，经硫酸钠干燥并过滤。真空浓缩滤液，得到黄色油状的粗品化合物(1.92g，100％)。LC-MS:ESI m/z 203.2[M+H] +；C 12H 14N 2O计算值202.11.
步骤3：3-(3,5-二甲基-1H-吡唑-1-基)苯甲醛
Figure PCTCN2021078537-appb-000138
在0℃，N 2气氛下，向(3-(3,5-二甲基-1H-吡唑-1-基)苯基)甲醇(1.92g，9.40mmol)的二氯甲烷(20mL)溶液中加入Dess-Martin高碘烷(11.0g，26.1mmol)。在25℃下搅拌16小时后，加入饱和Na 2S 2O 3和饱和NaHCO 3淬灭反应，并将所得混合物在25℃下搅拌直至不再放出气体。用水(20mL)稀释混合物，并用二氯甲烷(20mL×3)萃取。合并的有机相用盐水洗涤，硫酸钠干燥。滤液真空浓缩，得到标题化合物(1.9g，100％)，为黄色固体化合物。LC-MS:ESI m/z 201.2[M+H] +；C 12H 12N 2O计算值200.09.
步骤4.N-(3-(3,5-二甲基-1H-吡唑-1-基)亚苄基)-2-甲基丙烷-2-亚磺酰胺
Figure PCTCN2021078537-appb-000139
在N 2气氛下，向3-(3,5-二甲基-1H-吡唑-1-基)苯甲醛(1.9g，9.5mmol)和(R)-叔丁基亚磺酰胺(1.3g，11mmol)的DCE(20mL)溶液中加入CuSO 4(2.3g，14mmol)。在50℃下搅拌16小时后，将反应混合物冷却至室温，并通过硅藻土过滤，滤液用水和饱和食盐水洗涤。有机相经硫酸钠干燥并过滤，滤液减压浓缩后，通过层析柱纯化(0-60％乙酸乙酯的石油醚溶液)，得到黄色油状化合物(2.16g，75％)。 1H NMR(400MHz,DMSO-d 6)δ8.65(s,1H),8.04(s,1H),7.94(d,J＝7.6Hz,1H),7.73(d,J＝7.8Hz,1H),7.66(t,J＝7.8Hz,1H),6.11(s,1H),2.35(s,3H),2.19(s,3H),1.20(s,9H).LC-MS:ESI m/z 304.19[M+H] +；C 16H 21N 3OS计算值303.14.
步骤5：(S)-3-(((R)-叔丁基亚磺酰基)氨基)-3-(3-(3,5-二甲基-1H-吡唑-1-基)苯基)丙酸乙酯
Figure PCTCN2021078537-appb-000140
在N 2气氛下，将Zn(4.3g，66mmol)和CuCl(660mg，6.7mmol)加入到四氢呋喃(40mL)中，在75℃搅拌30分钟。然后移除加热浴，并缓慢加入溴乙酸乙酯(2mL，17mmol)的四氢呋喃(10mL)溶液。将反应混合物在25℃搅拌30分钟，然后升温到50℃搅拌30分钟。将反应混合物冷却至0℃，并将N-(3-(3,5-二甲基-1H-吡唑-1-基)亚苄基)-2-甲基丙烷-2-亚磺酰胺(2.0g 6.7mmol)的四氢呋喃(10mL)溶液滴加到反应液中。在0℃下搅拌3小时后，将反应液通过硅藻土过滤。滤液依次用1N盐酸水溶液，NaHCO 3水溶液，盐水洗涤，经硫酸钠干燥并过滤，真空除去滤液，得到呈黄色油状的粗品化合物(1.9g，76％)。 1H NMR(400MHz,DMSO-d 6)δ7.52–7.44(m,2H),7.42–7.35(m,2H),6.10(s,1H),5.72(d,J＝6.4Hz,1H),4.74(dd,J＝13.8,6.9Hz,1H),4.05(dd,J＝9.6,4.6Hz,2H),3.07(dd,J＝15.4,6.8Hz,1H),2.85(dd,J＝15.4,7.7Hz,1H),2.31(s,3H),2.21(s,3H),1.14(d,J＝7.2Hz,3H),1.11(s,9H).LC-MS:ESI m/z 392.3[M+H] +C 20H 29N 3O 3S计算值391.19.
步骤6：(S)-3-氨基-3-(3-(3,5-二甲基-1H-吡唑-1-基)苯基)丙酸乙酯
Figure PCTCN2021078537-appb-000141
将(S)-3-(((R)-叔丁基亚硫酰基)氨基)-3-(3-(3,5-二甲基-1H-吡唑-1-基)苯基)丙酸乙酯(1.9g 4.8mmol)溶于二氯甲烷(10mL)中，加入4N盐酸/1，4-二氧六环(5mL)。在25℃下搅拌16小时后，真空除去溶剂，得到黄色油状粗品标题化合物(1.6g，100％)。LC-MS:ESI m/z 288.2[M+H] +；C 16H 21N 3O 2计算值287.16.
步骤7：3-(((S)-1-(3-(3,5-二甲基-1H-吡唑-1-基)苯基)-3-乙氧基-3-氧丙基)氨基)吡咯烷-1-叔丁基羧酸酯
Figure PCTCN2021078537-appb-000142
在N 2气氛下，将(S)-3-氨基-3-(3-(3,5-二甲基-1H-吡唑-1-基)苯基)丙酸乙酯(1.3g，4.5mmol)和1-叔丁氧羰基-3-吡咯烷酮(1.7g，9.0mmol)溶于EtOH(20mL)中，加入AcOH(1.6mL)。在25℃下搅拌16小时后，将氰基硼氢化钠(567mg，9.0mmol)加入反应混合物中。在25℃下搅拌3小时后，真空除去溶剂。将残余物用饱和NaHCO 3水溶液稀释，然后用二氯甲烷(20mL×3)萃取。合并的有机相用盐水洗涤，经硫酸钠干燥并过滤。将滤液在减压下浓缩。将残余物通过柱层析纯化(0-60％乙酸乙酯的石油醚溶液)，得到白色油状化合物(960mg，80％)。LC-MS:ESI m/z 457.4[M+H] +；C 25H 36N 4O 4计算值456.27.
步骤8：(3S)-3-(3-(3,5-二甲基-1H-吡唑-1-基)苯基)-3-(吡咯烷-3-基氨基)丙酸乙酯
Figure PCTCN2021078537-appb-000143
将3-(((S)-1-(3-(3,5-二甲基-1H-吡唑-1-基)苯基)-3-乙氧基-3-氧丙基)氨基)吡咯烷-1-叔丁基羧酸酯(80mg，0.17mmol)溶于二氯甲烷(2mL)中，加入4N的盐酸/1，4-二氧六环(2mL)溶液。在25℃下搅拌16小时后，真空除去溶剂，得到黄色油状化合物(50mg)，其无需进一步纯化即可直接用于下一步。LC-MS:ESI m/z 357.2[M+H] +；C 20H 28N 4O 2计算值356.22.
步骤9：7-(2-(3-(((S)-1-(3-(3,5-二甲基-1H-吡唑-1-基)苯基)苯基)-3-乙氧基-3-氧丙基)氨基)吡咯烷-1-基)乙基)-3,4-二氢-1,8-萘啶-1(2H)-羧酸叔丁酯
Figure PCTCN2021078537-appb-000144
将(3S)-3-(3-(3,5-二甲基-1H-吡唑-1-基)苯基)-3-(吡咯烷-3-基氨基)丙酸乙酯(50mg，0.1mmol)和7-(2-(甲苯磺酰氧基)乙基)-3,4-二氢-1,8-萘啶-1(2H)-羧酸叔丁酯(52mg，0.12mmol)溶于乙腈(3mL)中，加入K 2CO 3(42mg，0.3mmol)和NaI(19mg,0.1mmol)。在N 2气氛下，于60℃搅拌16小时后，真空除去溶剂，并将粗残余物通过硅胶层析柱(0～30％甲醇的二氯甲烷溶液)，得到无色油状的标题化合物(24mg，48％)。LC-MS:ESI m/z 617.4[M+H] +；C 35H 48N 6O 4计算值616.37.
步骤10-11：(3S)-3-(3-(3,5-二甲基-1H-吡唑-1-基)苯基)-3-((1-(2-(5,6,7,8-四氢-1,8-萘啶-2-基)乙基)吡咯烷-3-基)氨基)丙酸
Figure PCTCN2021078537-appb-000145
将7-(2-(3-(((S)-1-(3-(3,5-二甲基-1H-吡唑-1-基)苯基)-3-乙氧基-3-氧丙基)氨基)吡咯烷-1-基)乙基)-3,4-二氢-1,8-萘啶-1(2H)-羧酸叔丁酯(24mg，0.04mmol)溶于甲醇(0.5mL)和H 2O(0.5mL)的混合溶剂中，加入LiOH(2mg，0.08mmol)。在25℃搅拌16小时后，真空除去溶剂，得到(3S)-3-((1-(2-(8-(叔丁氧羰基)-5,6,7,8-四氢-1,8-萘啶-2-基)乙基)吡咯烷-3-基)氨基)-3-(3-(3,5-二甲基-1H-吡唑-1-基)苯基)丙酸(23mg，98％)，为黄色固体。将(3S)-3-((1-(2-(8-(叔丁氧羰基)-5,6,7,8-四氢-1,8-萘啶-2-基)乙基)吡咯烷-3-基)氨基)-3-(3-(3,5-二甲基-1H-吡唑-1-基)苯基)丙酸(23mg，0.03mmol)溶于二氯甲烷(1mL)，加入4N的盐酸/1，4-二氧六环(1mL)。在25℃下搅拌3小时后，在真空下除去溶剂，将得到的粗品通过制备型反相HPLC纯化[色谱柱：Kromasil 100-5-C18，30×150mm；流动相：1-35％乙腈的水溶液(含0.1％甲酸)；时间：14分钟，]，得到白色固体的标题化合物(7.1mg，37％)。 1H NMR(400MHz,DMSO-d 6)δ7.57–7.41(m,4H),7.17–7.02(m,1H),6.37–6.29(m,1H),6.13(s,1H),4.17–4.11(m,1H),3.35–3.22(m,2H),3.16–3.03(m,1H),2.93–2.61(m,10H),2.52–2.43(m,2H),2.35(s,3H),2.24(s,3H),2.00–1.91(m,1H),1.85–1.71(m,3H).LC-MS:ESI m/z 489.42[M+H] +；C 28H 36N 6O 2计算值488.29.
参照化合物97的合成方法制备得到化合物98(甲酸盐)： 1H NMR(400MHz,DMSO-d 6)δ7.48–7.40(m,2H),7.36–7.31(m,1H),7.11(d,J＝7.9Hz,1H),7.07–6.95(m,1H),6.39–6.22(m,2H),6.07(s,1H),4.26(t,J＝7.5Hz,1H),3.24–3.20(m,2H),2.75–2.55(m,10H),2.28(d,J＝3.4Hz,5H),2.18(s,3H),2.07(s,3H),2.00–1.90(m,1H),1.80–1.66(m,3H).LC-MS:ESI m/z 503.33[M+H] +,C 29H 38N 6O 2计算值502.31
实施例18.化合物99的合成
Figure PCTCN2021078537-appb-000146
步骤1：3-(3-(3,5-二甲基-1H-吡唑-1-基)苯基)-3-氧代丙酸甲酯
Figure PCTCN2021078537-appb-000147
在N 2气氛下，向3-(3,5-二甲基-1H-吡唑-1-基)苯甲酸(10.0g，46.3mmol)的DMF(100mL)溶液中添加CDI(11.2g，69.4mmol)。在25℃下搅拌16小时后，将丙二酸单乙酯钾(18.0g，116mmol)和MgCl 2(8.8g，93mmol)加入反应混合物中。在50℃下搅拌16小时后，将反应混合物用水(300mL)稀释，并用乙酸乙酯(100mL×3)萃取。有机相用盐水洗涤，经硫酸钠干燥，并过滤。将滤液在真空下浓缩。残余物通过快速柱色谱纯化(0-30％乙酸乙酯的石油醚溶液)，得到标题化合物(3.3g，26％)，为黄色油状物。LC-MS:ESI m/z 273.16[M+H] +；C 15H 16N 2O 3计算值272.12.
步骤2：(3S)-3-((1-(3-(3,5-二甲基-1H-吡唑-1-基)苯基)-3-甲氧基-3-氧丙基)氨基)吡咯烷-1-羧酸叔丁基酯
Figure PCTCN2021078537-appb-000148
在N 2气氛下，向3-(3-(3,5-二甲基-1H-吡唑-1-基)苯基)-3-氧代丙酸甲酯(1.8g，6.6mmol)和(S)-3-氨基吡咯烷-1-羧酸叔丁酯(1.4g，7.9mmol)的甲醇(20mL)溶液中加入三乙酰氧基硼氢化钠(2.1g，9.9mmol)。在25℃下搅拌16小时后，将氰基硼氢化钠(624mg，9.9mmol)加入反应混合物中。在25℃下进一步搅拌16小时后，真空除去溶剂。将残余物用NaHCO 3(20mL)稀释，并用二氯甲烷(30mL×3)萃取。合并的有机相用盐水洗涤，经硫酸钠干燥，并过滤。将滤液在减压下浓缩。残余物通过快速柱色谱法纯化(0-100％乙酸乙酯的石油醚溶液)，得到标题化合物(873mg，30％)，为无色油状物。 1H NMR(400MHz,CDCl 3)δ7.47–7.41(m,2H),7.39–7.32(m,2H),6.01(s,1H),4.23(br,1H),3.67(s,3H),3.56–3.38(m,2H),3.36–3.19(m,2H),3.18–3.14(m,1H),2.71–2.68(m,2H),2.31(s,3H),2.29(s,3H),2.04–1.97(m,2H),1.48–1.41(m,9H).LC-MS:ESI m/z 443.24[M+H] +；C 24H 34N 4O 4计算值442.26.
步骤3：3-(3-(3,5-二甲基-1H-吡唑-1-基)苯基)-3-(((S)-吡咯烷-3-基)氨基)丙酸甲酯
Figure PCTCN2021078537-appb-000149
向(3S)-3-((1-(3-(3,5-二甲基-1H-吡唑-1-基)苯基)-3-甲氧基-3-氧丙基)氨基)吡咯烷-1-甲酸叔丁酯(873mg，1.9mmol)的二氯甲烷(8mL)溶液中加入TFA(2mL)。在25℃下搅拌2小时后，真空除去溶剂，得到标题化合物(892mg，粗品)，为无色油状的TFA盐形式，其无需进一步纯化即可用于下一步。LC-MS:ESI m/z 343.2[M+H] +；C 19H 26N 4O 2计算值342.21.
步骤4：7-(2-((3S)-3-((1-(3-(3,5-二甲基-1H-吡唑-1-基)苯基)-3--3-甲氧基)氨基)吡咯烷-1-基)乙基)-3,4-二氢-1,8-萘啶-1(2H)-羧酸叔丁基酯
Figure PCTCN2021078537-appb-000150
在N 2气氛下，向TFA盐形式的3-(3-(3,5-二甲基-1H-吡唑-1-基)苯基)-3-(((S)-吡咯烷-3-基)氨基)丙酸甲酯(892mg，1.9mmol)和7-(2-(甲苯磺酰氧基)乙基)-3,4-二氢-1,8-萘啶-1(2H)-羧酸叔丁酯(1.0g，2.4mmol)的乙腈(15mL)溶液中加入K 2CO 3(1.3g，9.5mmol)和NaI(353mg，1.9)。在70℃搅拌16小时后，真空除去溶剂，并将粗残余物通过快速柱色谱纯化(0～30％甲醇的二氯甲烷溶液)得到标题化合物(727mg，66％)，为白色固体。LC-MS:ESI m/z 603.4[M+H] +；C 34H 46N 6O 4计算值602.36.
步骤5：3-(3-(3,5-二甲基-1H-吡唑-1-基)苯基)-3-(((S)-1-(2-(5,6,7,8-四氢-1,8-萘啶-2-基)乙基吡咯烷-3-基)氨基)丙酸
Figure PCTCN2021078537-appb-000151
向7-(2-((3S)-3-((1-(3-(3,5-二甲基-1H-吡唑-1-基)苯基)-3-甲氧基-3-氧丙基)氨基)吡咯烷-1-基)乙基)-3,4-二氢-1,8-萘啶-1(2H)-羧酸叔丁基酯(727mg，1.2mmol)的二氯甲烷(6mL)溶液中加入TFA(2mL)。在25℃下搅拌2小时后，真空除去溶剂，得到TFA盐形式的标题化合物(735mg，粗品)。将TFA盐(735mg，1.2mmol)形式的甲基3-(3-(3,5-二甲基-1H-吡唑-1-基)苯基)-3-(((S)-1-(2-(5,6,7,8-四氢-1,8-萘啶-2-基)乙基吡咯烷-3-基)氨基)丙酸酯溶到甲醇(4mL)和H 2O(1mL)的混合溶剂中，后加入NaOH(96mg，2.4mmol)。在25℃下搅拌3小时后，在真空下除去溶剂，并在以下条件下通过制备的手性SFC 分离粗残余物[柱：Daicel Chiralpak IG(250mm×30mm，10μm)；流动相：60％甲醇的二氧化碳溶液(0.1％NH 3.H 2O)；流速：70g/min]，分别得到化合物99a和化合物99b，为白色固体。化合物99a：(85mg，14％)，白色固体。 1H NMR(400MHz,CD 3OD)δ7.51–7.43(m,3H),7.36–7.31(m,1H),7.15(d,J＝7.3Hz,1H),6.36(d,J＝7.3Hz,1H),6.06(s,1H),4.24(dd,J＝9.3,4.8Hz,1H),3.70–3.62(m,1H),3.49–3.35(m,5H),3.18–3.03(m,3H),2.88–2.78(m,2H),2.71(t,J＝6.2Hz,2H),2.60–2.49(m,2H),2.28(s,3H),2.24(s,3H),2.23–2.12(m,2H),1.92–1.85(m,2H).LC-MS:ESI m/z 489.3[M+H] +；C 28H 36N 6O 2计算值488.29.采用分析型手性SFC(Chiralpak IG-3(50×4.6mm)柱子)得到保留时间：2.2min，ee值：99.5％.
化合物99b：(80mg，13％)，白色固体。 1H NMR(400MHz,CD 3OD)δ7.53–7.45(m,3H),7.39–7.29(m,1H),7.20(d,J＝7.3Hz,1H),6.43(d,J＝7.3Hz,1H),6.09(s,1H),4.19(dd,J＝8.3,5.8Hz,1H),3.58(d,J＝8.0Hz,1H),3.50–3.46(m,1H),3.40–3.34(m,5H),3.16–3.05(m,2H),2.94(t,J＝6.4Hz,2H),2.72(t,J＝6.2Hz,2H),2.61–2.51(m,2H),2.29(s,3H),2.26(s,3H),2.22–2.15(m,1H),1.92–1.82(m,3H).LC-MS:ESI m/z 489.3[M+H] +；C 28H 36N 6O 2计算值488.29.采用分析型手性SFC(Chiralpak IG-3(50×4.6mm)柱子)得到保留时间：1.3min，ee值：100.0％.
实施例19.参照化合物99的合成，制备得到以下化合物：
化合物100(甲酸盐): 1H NMR(400MHz,CD 3OD)δ7.59–7.29(m,5H),6.61–6.53(m,0.5H),6.49–6.38(m,0.5H),6.09(s,1H),4.40–4.30(m,1H),4.17–3.97(m,1H),3.94–3.76(m,1H),3.70–3.52(m,3H),3.49–3.36(m,4H),2.83–2.54(m,4H),2.26(s,3H),2.25(s,3H),2.19–1.98(m,2H),1.97–1.84(m,2H).LC-MS:ESI m/z 503.2[M+H] +；C 28H 34N 6O 3.计算值502.27.化合物101(甲酸盐)： 1H NMR(400MHz,CD 3OD)δ7.62–7.47(m,5H),6.55(d,J＝7.3Hz,1H),6.10(s,1H),4.60(d,J＝10.1Hz,1H),3.77–3.68(m,1H),3.58–3.41(m,3H),3.37–3.32(m,1H),3.15–3.08(m,1H),2.98–2.41(m,11H),2.31(s,3H),2.24(s,3H),2.04–1.80(m,4H).LC-MS:ESI m/z 503.3[M+H] +；C 29H 38N 6O 2计算值502.31.
化合物102(甲酸盐)： 1H NMR(400MHz,CD 3OD)δ7.59–7.34(m,5H),6.63–6.46(m,1H),6.11–6.06(m,1H),4.47–4.28(m,1.5H),3.99(m,0.5H),3.76–3.65(m,1H),3.64–3.47(m,2H),3.46–3.33(m,4H),3.15–3.02(m,1H),2.98–2.82(m,2H),2.82–2.70(m,2H),2.69–2.56(m,2H),2.32–2.21(m,6H),2.15–1.99(m,2H),1.96–1.86(m,2H).LC-MS:ESI m/z 517.2[M+H] +；C 29H 36N 6O 3计算值516.28.
化合物103(甲酸盐)： 1H NMR(400MHz,CD 3OD)δ7.70–7.36(m,5H),6.51(d,J＝7.3Hz,1H),6.10(s,1H),4.56(d,J＝8.8Hz,1H),3.73(d,J＝11.3Hz,1H),3.50–3.35(m,4H),3.33-3.32(m,1H),3.02–2.63(m,8H),2.56–2.42(m,2H),2.31(s,3H),2.26(s,3H),2.16–2.06(m,1H),1.96–1.61(m,6H).LC-MS:ESI m/z 517.3[M+H] +；C 30H 40N 6O 2计算值516.32.
化合物104(甲酸盐)： 1H NMR(400MHz,CD 3OD)δ7.55–7.33(m,5H),6.54–6.45(m,1H),6.08(s,1H),4.28–4.15(m,1H),4.07–3.96(m,1H),3.85–3.75(m,1H),3.75–3.61(m,2H),3.54–3.38(m,4H),3.21–3.11(m,1H),2.82–2.67(m,5H),2.64–2.46(m,3H),2.26(s,3H),2.25(s,3H),1.99–1.84(m,4H).LC-MS:ESI m/z 531.1[M+H] +；C 30H 38N 6O 3.计算值530.30.
制备得到化合物105，并通过制备手性SFC，在以下条件下分离粗品：[柱：Daicel ChiralPak IG(250×30mm，10μm)；流动相：60％甲醇的二氧化碳溶液(0.1％NH 3.H 2O)；流速：70g/min]，分别得到化合物105a和化合物105b，为白色固体。
化合物105a： 1H NMR(400MHz,CD 3OD)δ7.48–7.42(m,3H),7.33–7.29(m,1H),7.13(d,J＝7.3Hz,1H),6.33(d,J＝7.3Hz,1H),6.04(s,1H),4.21(dd,J＝9.3,4.8Hz,1H),3.68–3.61(m,1H),3.47–3.32(m,5H),3.16–3.01(m,3H),2.85–2.77(m,2H),2.69(t,J＝6.2Hz,2H),2.58–2.47(m,2H),2.24(s,3H),2.23(s,3H),2.21–2.12(m,2H),1.89–1.82(m,2H).LC-MS:ESI m/z 489.1[M+H] +；C 28H 36N 6O 2计算值488.29.采用分析型手性SFC(Chiralpak IG-3(50×4.6mm)柱子)得到保留时间：1.3min，ee值：98.4％.
化合物105b： 1H NMR(400MHz,CD 3OD)δ7.52–7.43(m,3H),7.35–7.29(m,1H),7.18(d,J＝7.3Hz,1H),6.41(d,J＝7.3Hz,1H),6.06(s,1H),4.17(dd,J＝8.2,5.9Hz,1H),3.59(d,J＝8.0Hz,1H),3.52–3.45(m,1H),3.44–3.33(m,5H),3.14–3.02(m,2H),2.92(t,J＝6.3Hz,2H),2.70(t,J＝6.1Hz,2H),2.59–2.48(m,2H),2.27(s,3H),2.24(s,3H),2.21–2.13(m,1H),1.92–1.79(m,3H).LC-MS:ESI m/z 489.5[M+H] +；C 28H 36N 6O 2计算值488.29.采用分析型手性SFC(Chiralpak IG-3(50×4.6mm)柱子)得到保留时间：2.9min，ee值：100.0％.
化合物106(甲酸盐)： 1H NMR(400MHz,CD 3OD)δ7.54–7.34(m,5H),6.54–6.47(m,1H),6.08(s,1H),4.34–4.24(m,1H),3.56–3.53(m,1H),3.43–3.41(m,4H),3.13–3.04(m,4H),2.78–2.75(m,2H),2.61–2.55(m,4H),2.29(s,3H),2.25(s,3H),2.21–2.04(m,2H),1.92–1.90(m,2H),1.78–1.66(m,4H),1.48–1.42(m,2H).LC-MS:ESI m/z 531.3[M+H] +；C 31H 42N 6O 2计算值530.34.
化合物107(甲酸盐)： 1H NMR(400MHz,CD 3OD)δ7.57–7.33(m,5H),6.55–6.46(m,1H),6.08(s,1H),4.45–4.25(m,1.5H),3.98–3.90(m,0.5H),3.82–3.36(m,7H),3.29–3.12(m,3H),2.91–2.74(m,3H),2.71–2.42(m,5H),2.25(s,3H),2.22(s,3H),1.99–1.86(m,2H),1.80–1.64(m,2H).LC-MS:ESI m/z 545.3[M+H] +；C 31H 40N 6O 3计算值544.32.
实施例20 化合物108的合成
Figure PCTCN2021078537-appb-000152
步骤1：3-羟基-2-苯基丙酸甲酯
Figure PCTCN2021078537-appb-000153
向2-苯基乙酸甲酯(10.0g，66.6mmol)的DMSO(150mL)溶液中加入甲醛(2.1g，70mmol)和叔丁醇钾(1.5g，13.33mmol)，反应液于室温下反应16小时，后将冰水(100mL)倒入 反应液中，滴加1N盐酸调节pH≈5，所得混合物用水(100mL)稀释，乙酸乙酯(150mL×2)萃取，合并的有机相用饱和NaHCO 3(20mL)和盐水洗涤，经硫酸钠干燥，残余物通过快速柱色谱法纯化(0-15％乙酸乙酯的石油醚溶液)，得到标题化合物(7.0g，58％)，为黄色油状物。LC-MS:ESI m/z 181.2[M+H] +,C 10H 12O 3计算值180.08.
步骤2：2-苯基丙烯酸甲酯
Figure PCTCN2021078537-appb-000154
在0℃，N 2气氛下，向3-羟基-2-苯基丙酸甲酯(5.0g，27.8mmol)和三乙胺(7.0g，69.5mmol)的二氯甲烷(60mL)溶液中加入甲磺酰氯(3.5g，30.47mmol)，升温到室温下搅拌14小时。用饱和NaHCO 3(80mL)淬灭反应，二氯甲烷(80mL×2)萃取。合并的有机相用盐水洗涤，经硫酸钠干燥，过滤并浓缩至干。残余物通过快速柱色谱法纯化(0-40％乙酸乙酯的石油醚溶液)，得到标题化合物(2.7g，60％)，为黄色油状物。 1H NMR(400MHz,DMSO-d 6)δ7.49-7.33(m,5H),6.25(s,1H),6.03(s,1H),3.76(s,3H).LC-MS:ESI m/z 163.2[M+H] +,C 10H 10O 2计算值162.07.
步骤3：(3R)-3-((3-甲氧基-3-氧代-2-苯基丙基)氨基)吡咯烷-1-羧酸叔丁酯
Figure PCTCN2021078537-appb-000155
将2-苯基丙烯酸甲酯(800mg，4.9mmol)，(R)-3-氨基吡咯烷-1-甲酸叔丁酯(1.4g，7.35mmol)加入到四氢呋喃(8mL)中，反应液在25℃下搅拌16小时。真空除去溶剂。残余物用水稀释，并用乙酸乙酯(15mL×2)萃取。合并的有机相用盐水洗涤，经硫酸钠干燥，过滤并浓缩至干。残余物通过快速柱色谱法纯化(0-50％乙酸乙酯在石油醚中)，得到标题化合物(1.06g，62％)，为黄色固体。LC-MS:ESI m/z 349.5[M+H] +,C 19H 28N 2O 4计算值348.20.
步骤4-7参照化合物97步骤9-11的合成方法进行，制备得到化合物108(甲酸盐)： 1HNMR(400MHz,CD 3OD)δ7.33-7.28(m,5H),7.26-7.24(m,1H),6.51-6.49(m,1H),3.79-3.77(m,1H),3.74-3.71(m,1H),3.47-3.42(m,2H),3.40-3.33(m,2H),3.32-3.30(m,2H),3.27-3.11(m,2H),3.04-3.01(m,2H),2.87-2.58(m,4H),2.31–2.30(m,1H),2.27-2.03(m,1H),1.96-1.85(m,2H).LC-MS:ESI m/z 395.1[M+H] +,C 23H 30N 4O 2.计算值394.24.
实施例21.参照化合物108的合成方法，制备得到以下化合物：
化合物109(甲酸盐)： 1H NMR(400MHz,CD 3OD)δ7.36–7.32(m,1H),7.36-7.30(m,5H),6.59-6.58(m,1H),4.02-4.00(m,1H),3.76-3.73(m,2H),3.65-3.54(m,1H),3.43-3.30(m,2H),3.12-2.92(m,4H),2.91-2.78(m,4H),2.40-1.95(m,4H),1.93-1.88(m,2H).LC-MS:ESI m/z 395.1[M+H] +；C 23H 30N 4O 2.计算值394.24.
化合物110(甲酸盐)：1H NMR(400MHz,CD 3OD)δ7.87(t,J＝7.5Hz,2H),7.60–7.53(m,1H),7.52–7.45(m,3H),6.59–6.54(m,1H),4.61–4.55(m,1H),3.79–3.73(m,1H),3.59–3.32(m,5H),3.25–2.96(m,3H),2.91–2.88(m,2H),2.79–2.75(m,2H),2.61–2.48(m,2H),2.39–2.30(m,1H),2.03–1.94(m,1H),1.93–1.90(m,2H).LC-MS:ESI m/z 438.2[M+H] +； C 24H 31N 5O 3计算值437.24.
化合物111(甲酸盐)： 1H NMR(400MHz,CD 3OD)δ7.87(t,J＝7.5Hz,2H),7.60–7.55(m,1H),7.52–7.46(m,3H),6.59–6.56(m,1H),4.60–4.54(m,1H),3.79–3.75(m,1H),3.45–3.30(m,5H),3.23–2.93(m,3H),2.93–2.89(m,2H),2.82–2.76(m,2H),2.59–2.49(m,2H),2.43–2.33(m,1H),2.03–1.97(m,1H),1.93–1.89(m,2H).LC-MS:ESI m/z 438.2[M+H] +；C 24H 31N 5O 3计算值437.24.
实施例22.化合物112的合成
Figure PCTCN2021078537-appb-000156
步骤1：3-((3-乙氧基-3-氧代丙-1-烯-1-基)氧基)吡咯烷-1-甲酸叔丁酯
Figure PCTCN2021078537-appb-000157
在25℃下，向(3-羟基吡咯烷-1-基)甲酸叔丁酯(10g，53mmol)的无水二氯甲烷(15mL)溶液中加入N-甲基吗啉(5.9g，58mmol)和丙炔酸乙酯(5.7g，58mmol)。将反应混合物搅拌4小时。将混合物浓缩至干。将粗产物在硅胶柱上使用乙酸乙酯-己烷(1：1)纯化，得到呈黄色油状的标题化合物(15g，91％)。 1H NMR(400MHz,CDCl 3)δ7.51(d,J＝12.8Hz,1H),5.21(d,J＝12.8Hz,1H),4.62(s,1H),4.15(t,J＝7.2Hz,2H),3.57–3.40(m,4H),2.20–2.04(m,2H),1.47(s,9H),1.28(t,J＝7.2Hz,3H).LC-MS:ESI m/z 308.3[M+Na] +；C 14H 23NO 5计算值285.16.
步骤2：3-((1-(3-(3,5-二甲基-1H-吡唑-1-基)苯基)-3-乙氧基-3-氧代丙-1-烯-1-基)氧基)吡咯烷-1-羧酸叔丁酯
Figure PCTCN2021078537-appb-000158
在N 2气氛下，向1-(3-溴苯基)-3,5-二甲基吡唑(1.0g，4.0mmol)的1，4-二氧六环(10mL)溶液加入3-((3-乙氧基-3-氧代丙-1-烯-1-基)氧基)吡咯烷-1-羧酸叔丁酯(1.7g，6.0mmol)，LiCl(0.51g，12mmol)，N，N-二环己基甲胺(86mg，0.4mmol)，双(三叔丁基膦)钯(0) (0.20g，0.40mmol)，反应液在110℃下搅拌14小时。将该混合物倒入水(20mL)中淬灭，并用乙酸乙酯(20mL×3)萃取。合并的有机相用饱和食盐水洗涤，经硫酸钠干燥并浓缩至干。粗产物通过FCC(0～20％乙酸乙酯的石油醚溶液)纯化，得到棕色胶状的标题化合物(0.34g，17％)。 1H NMR(400MHz,CDCl 3)δ7.64–7.34(m,4H),6.08–5.94(m,1H),4.96–4.80(m,1H),4.28–3.97(m,3H),3.78–3.32(m,4H),2.37–2.28(m,6H),1.76–1.53(m,2H),1.52–1.46(m,9H),1.35–1.22(m,3H).LC-MS:ESI m/z 456.3[M+H] +；C 25H 33N 3O 5计算值455.24.
步骤3：3-(1-(3-(3,5-二甲基-1H-吡唑-1-基)苯基)-3-乙氧基-3-氧丙氧基)吡咯烷-1-甲酸叔丁酯
Figure PCTCN2021078537-appb-000159
向3-((1-(3-(3,5-二甲基-1H-吡唑-1-基)苯基)-3-乙氧基-3-氧代丙-1-烯-1-基)氧基)吡咯烷-1-羧酸叔丁酯(0.28g，0.66mmol)的乙酸乙酯(10mL)溶液中加入10％的Pd/C(28mg)，反应液在1大气压的H 2气氛下于80℃下搅拌5小时。过滤混合物并浓缩，得到3-(1-(3-(3,5-二甲基-1H-吡唑-1-基)苯基)-3-乙氧基-3-氧代丙氧基)吡咯烷-1-羧酸叔丁酯(0.30g，96％)，为黄色油状物，其无需进一步纯化即可用于下一步。 1H NMR(400MHz,CDCl 3)δ7.48–7.32(m,4H),6.01(s,1H),4.93–4.82(m,1H),4.18–3.98(m,3H),3.41–3.19(m,4H),2.79–2.74(m,1H),2.60–2.53(m,1H),2.31–2.30(m,6H),1.87–1.74(m,2H),1.48–1.42(m,9H),1.28–1.24(m,3H).LC-MS:ESI m/z 458.3[M+H] +；C 25H 35N 3O 5计算值457.26.
步骤4-7：参照化合物23的合成(步骤2-5)进行，制备得到化合物112： 1H NMR(400MHz,DMSO-d 6,80℃)δ7.48–7.42(m,2H),7.39–7.33(m,2H),7.03–6.97(m,1H),6.31–6.23(m,1H),6.05(s,1H),5.94(brs,1H),4.84–4.76(m,1H),3.95(s,1H),3.31–3.22(m,2H),2.77–2.53(m,10H),2.47–2.33(m,2H),2.29(s,3H),2.19(s,3H),1.86–1.74(m,3H),1.59–1.57(m,1H).LC-MS:ESI m/z 490.3[M+H] +；C 28H 35N 5O 3计算值489.27.HPLC纯度:100.0％(214nm),100.0％(254nm).
实施例23.化合物113的合成
Figure PCTCN2021078537-appb-000160
步骤1：3-氯-3-苯基丙烯酸甲酯
Figure PCTCN2021078537-appb-000161
向氯化铝(2.0g，16mmol)，二氯甲烷(10mL)和环己烷(2.6g，31mmol)的混合液中加入苯丙炔酸甲酯(0.30mmol，0.48g)。将反应混合物在25℃下搅拌15小时。将该混合物倒入冰水(50mL)中，并用CH 2Cl 2(50mL×2)萃取。合并的有机相用饱和食盐水(30mL×3)洗涤，用无水硫酸钠干燥并浓缩至干。通过硅胶上的柱色谱法(石油醚/乙酸乙酯＝25：1)纯化粗产物，得到3-氯-3-苯基丙烯酸甲酯(0.50g，77％)，为黄色油状物。 1H NMR(400MHz,CDCl 3)δ7.46–7.39(m,5H),6.35(s,1H),3.60(s,3H).LC-MS:ESI m/z 197.1,199.1[M+H,Cl] +；C 10H 9ClO 2计算值196.03.
步骤2：3-((3-甲氧基-3-氧代-1-苯基丙-1-烯-1-基)氨基)-3-甲基吡咯烷-1-羧酸叔丁酯
Figure PCTCN2021078537-appb-000162
将密封管中的3-氯-3-苯基丙烯酸甲酯(0.88g，4.5mmol)和3-氨基-3-甲基吡咯烷-1-羧酸叔丁酯(0.30g，1.5mmol)的混合物在150℃下搅拌3小时。将混合物浓缩至干，得到3-((3-甲氧基-3-氧代-1-苯基丙-1-烯-1-基)氨基)-3-甲基吡咯烷-1-甲酸叔丁酯(1.1g，粗品)，为红色油状物，其无需进一步纯化直接用于下一步。LC-MS:ESI m/z 361.1[M+H] +；C 20H 28N 2O 4计算值360.20.
步骤3：3-((3-甲氧基-3-氧代-1-苯基丙基)氨基)-3-甲基吡咯烷-1-羧酸叔丁酯
Figure PCTCN2021078537-appb-000163
在25℃下，向3-((3-甲氧基-3-氧代-1-苯基丙-1-烯-1-基)氨基)-3-甲基吡咯烷-1-羧酸叔丁酯(0.80g，粗品)和HOAc(0.13g，0.22mmol)的甲醇(20mL)溶液中分批加入氰基硼氢化钠(0.28g，4.4mmol)。将反应混合液在25℃下搅拌2小时。然后用饱和NaHCO 3(50mL)淬灭反应，并用二氯甲烷(30mL×2)萃取。合并的有机相用盐水(50mL)洗涤，经硫酸钠干燥并浓缩，得到3-((3-甲氧基-3-氧代-1-苯基丙基)氨基)-3-甲基吡咯烷-1-羧酸叔丁酯(粗品为0.90g，为红色油状物，无需进一步纯化即可用于下一步。LC-MS:ESI m/z 363.3[M+H] +；C 20H 30N 2O 4计算值362.22.
步骤4：3-((3-甲基吡咯烷-3-基)氨基)-3-苯基丙酸甲酯
Figure PCTCN2021078537-appb-000164
向3-((3-甲氧基-3-氧代-1-苯基丙基)氨基)-3-甲基吡咯烷-1-羧酸叔丁酯(0.80g，粗品)在甲醇(15mL)中的溶液中加入盐酸/1，4-二氧六环(5.4mL，22mmol)。将混合物在25℃下搅拌18小时。浓缩混合物，得到红色油状3-((3-甲基吡咯烷-3-基)氨基)-3-苯基丙酸甲酯(0.40g，粗品)，其无需进一步纯化即可用于下一步。LC-MS:ESI m/z 263.1[M+H] +；C 15H 22N 2O 2计算值262.17.
步骤5：7-(2-(3-((3-甲氧基-3-氧代-1-苯基丙基)氨基)-3-甲基吡咯烷-1-基)乙基)-3,4-二氢-1，8-萘啶-1(2H)-羧酸叔丁酯
Figure PCTCN2021078537-appb-000165
在25℃，N 2下，向7-(2-碘乙基)-3,4-二氢-1,8-萘啶-1(2H)-羧酸叔丁酯(0.20g，0.51mmol)和3-((3-甲基吡咯烷-3-基)氨基)-3-苯基丙酸甲酯(0.27g，粗品)在乙腈(5mL)中的溶液中分批加入DIPEA(0.33g，1.53mmol)。将反应混合物在25℃下搅拌18小时。用水(50mL)淬灭混合物，并用二氯甲烷(30mL×2)萃取。合并的有机相用盐水(50mL)洗涤，经硫酸钠干燥并浓缩至干。粗产物通过硅胶柱色谱法(1～5％甲醇的二氯甲烷溶液)纯化，得到白色固体状的7-(2-(3-((3-甲氧基-3-氧代-1-苯基丙基)氨基)-3-)甲基吡咯烷-1-基)乙基)-3,4-二氢-1,8-萘啶-1(2H)-羧酸叔丁酯(0.15g，42％)。LC-MS:ESI m/z 523.3[M+H] +；C 30H 42N 4O 4计算值522.32.
步骤6：甲基3-((3-甲基-1-(2-(5,6,7,8-四氢-1,8-萘啶-2-基)乙基)吡咯烷-3-基)氨基)-3-苯丙酸酯
Figure PCTCN2021078537-appb-000166
向7-(2-(3-((3-甲氧基-3-氧代-1-苯基丙基)氨基)-3-甲基吡咯烷-1-基)乙基)-3,4-二氢-1,8-萘啶-1(2H)-羧酸叔丁酯(0.15g，0.29mmol)在甲醇(5mL)中的溶液中加入盐酸/1，4-二氧六环(0.8mL，2.9mmol)。将混合物在25℃下搅拌18小时。浓缩混合物，得到甲基3-((3-甲基-1-(2-(5,6,7,8-四氢-1,8-萘啶-2-基)乙基)吡咯烷-3-基)氨基)-3-苯基丙酸酯(0.18g，粗品)，为红色固体，其无需进一步纯化即可用于下一步。LC-MS:ESI m/z 423.3[M+H] +；C 25H 34N 4O 2计算值422.27.
步骤7：3-((3-甲基-1-(2-(5,6,7,8-四氢-1,8-萘啶-2-基)乙基)吡咯烷-3-基)氨基)-3-苯丙酸
Figure PCTCN2021078537-appb-000167
向3-((3-甲基-1-(2-(5,6,7,8-四氢-1,8-萘啶-2-基)乙基)吡咯烷基-3-基)氨基)-3-苯基丙酸甲酯(0.32g，0.62mmol)的甲醇/H 2O(8mL，V/V＝10∶1)溶液中分批加入LiOH·H 2O(57mg，3.1mmol)。将混合物在25℃下搅拌18小时。然后用盐酸(1N)酸化至pH＝4～5，并浓缩至干。粗产物通过制备型HPLC(柱：Xbridge 5u C18 150×19mm；流动相：乙腈-水(0.05％NH 3·H 2O)；梯度：5～15％乙腈；流速：20mL/min)纯化得到标题化合物(6mg，4.3％)，为黄色固体。 1H NMR(400MHz,DMSO-d 6,80℃)δ7.46–7.34(m,2H),7.31–7.26(m,2H),7.25–7.19(m,1H),7.04–6.99(m,1H),6.28–6.24(m,1H),5.96(brs,1H),4.23–4.15(m,1H),3.27–3.25(m,2H),2.75–2.63(m,1H),2.62–2.54(m,4H),2.47–2.40(m,4H),2.34–2.05(m,2H),1.90(s,1H),1.84–1.37(m,4H),1.07(s,1.5H),1.05(s,1.5H).LC-MS:ESI m/z 409.3[M+H] +；C 24H 32N 4O 2计算值408.25.HPLC纯度:100.0％(214nm),100.0％(254nm).
实施例24.化合物114的合成
Figure PCTCN2021078537-appb-000168
步骤1：3-(((R)-2-甲氧基-2-氧代-1-苯基乙基)氨基甲酰基)吡咯烷-1-羧酸叔丁酯
Figure PCTCN2021078537-appb-000169
在25℃下，向1-(叔丁氧基羰基)吡咯烷-3-羧酸(3.5g，24mmol)，1-羟基苯并三唑(3.3g，16mmol)和N-(3-二甲基氨基丙基)-N'-乙基碳二亚胺盐酸盐(4.6g，24mmol)在二氯甲烷(40mL)的溶液中，加入三乙胺(4.9g，49mmol)和(R)-2-氨基-2-苯基乙酸甲酯(4.9g，24mmol)。将得到的混合物在25℃搅拌14小时。用水(60mL)淬灭反应，并用二氯甲烷(80mL×2)萃取。有机相用NH 4Cl水溶液(80mL×2)和盐水(100mL)洗涤，经硫酸钠干燥，过滤并浓缩。残余物通过硅胶柱色谱法纯化(二氯甲烷：甲醇＝60：1-20：1)，得到黄色油状物的标题化合物(6.0g，99％)。 1H NMR(400MHz,DMSO-d 6)δ8.88–8.81(m,1H),7.43–7.33(m,5H),5.41–5.39(d,J＝7.0Hz,1H),3.62(s,3H),3.49–3.35(m,2H),3.30–3.17(m,2H),3.13–3.09(m,1H),2.03–1.89(m,2H),1.40–1.38(m,9H).LC-MS:(ESI)m/z 307.2[M-55] +,C 19H 26N 2O 5计算值362.18.
步骤2：(2R)-2-苯基-2-(吡咯烷-3-甲酰胺基)乙酸甲酯盐酸盐
Figure PCTCN2021078537-appb-000170
向3-(((R)-2-甲氧基-2-氧代-1-苯基乙基)氨基甲酰基)吡咯烷-1-羧酸叔丁酯(6.0g，16mmol)的甲醇(15mL)溶液中加入4N的盐酸/1，4-二氧六环(15mL)溶液。将混合物在50℃下搅拌3小时。浓缩混合物，得到标题化合物(4.1g，94％)，为白色固体。LC-MS:(ESI)m/z263.3[M+1] +,C 14H 18N 2O 3计算值262.13.
步骤3：7-(2-(3-(((R)-2-甲氧基-2-氧代-1-苯基乙基)氨基甲酰基)吡咯烷-1-基)-2-氧代乙基)-3,4-二氢-1,8-萘啶-1(2H)-羧酸叔丁酯
Figure PCTCN2021078537-appb-000171
在25℃下，向2-(8-(叔丁氧羰基)-5,6,7,8-四氢-1,8-萘啶-2-基)乙酸(1.0g，3.4mmol)和1-羟基苯并三唑(0.68g，5.9mol)，N-(3-二甲基氨基丙基)-N’-乙基碳二亚胺盐酸盐(0.98g，5.9mmol)的DMF(20mL)溶液中加入三乙胺(1.0g，10mmol)和(R)-2-氨基-2-苯基乙酸甲酯(1.0g，4.0mmol)。将反应液在25℃搅14小时。用水(80mL)淬灭反应，并用二氯甲烷(100mL×2)萃取。有机相用NH 4Cl水溶液(50mL×2)和盐水(100mL)洗涤，经硫酸钠干燥，过滤并浓缩。将残余物通过硅胶柱色谱法(二氯甲烷：甲醇＝60:1-30:1)纯化，得到标题化合物(0.49g，11％)。LC-MS:(ESI)m/z 537.2[M+H] +,C 29H 36N 4O 6计算值536.26.
步骤4：(2R)-2-苯基-2-(1-(2-(5,6,7,8-四氢-1,8-萘啶-2-基)乙酰基)吡咯烷-3-甲酰胺基)乙酸甲酯盐酸盐
Figure PCTCN2021078537-appb-000172
向7-(2-(3-(((R)-2-甲氧基-2-氧-1-苯基乙基)氨基甲酰基)吡咯烷-1-基)-2-氧乙基)-3,4-二氢-1，8-萘啶-1(2H)-羧酸叔丁酯(0.49g，0.91mmol)的甲醇(2.0mL)溶液中加入4N的盐酸/1，4-二氧六环(6.0mL)溶液。将混合物在50℃下搅拌3小时。浓缩混合物得到呈黄色固体状标题化合物(0.56g，粗品)。LC-MS:(ESI)m/z 437.2[M+H] +,C 24H 28N 4O 4计算值436.21.
步骤5：(2R)-2-苯基-2-(1-(2-(5,6,7,8-四氢-1,8-萘啶-2-基)乙酰基)吡咯烷-3-甲酰胺基)乙酸
Figure PCTCN2021078537-appb-000173
在0℃下，向(2R)-2-苯基-2-(1-(2-(5-(6,7,8-四氢-1,8-萘啶-2-基)乙酰基)吡咯烷-3-甲酰胺基)乙酸甲酯盐酸盐(0.56g，1.2mmol)溶解于甲醇(6.0mL)和四氢呋喃(2.0mL)的溶液中，然后滴加2N的LiOH(3.3mL，6.5mmol)。将反应液升温到室温并反应2小时。将混合物用1N盐酸酸化直至pH＝7-8。将水相冻干，得到的粗品通过制备型HPLC纯化(色谱柱：Kromasil-C18 100×21.2mm 5um；流动相：乙腈-水(0.05％NH 3.H 2O)；梯度：10-20％；流速：20mL/min)得到白色固体状的标题化合物(8.9mg，2％)。 1H NMR(400MHz,DMSO-d 6,80℃)δ7.39–7.26(m,5H),7.05-7.03(m,1H),6.33–6.28(m,1H),6.15–5.87(brs,1H),5.29(s,1H),3.73–3.61(m,2H),3.62–3.50(m,2H),3.47–3.44(m,3H),3.18–3.06(m,2H),2.68–2.58(m,2H),2.05–1.91(m,2H),1.82–1.72(m,2H).LC-MS:(ESI)m/z 423.2[M+H] +；C 23H 26N 4O 4计算值422.20.HPLC纯度:95.8％(214nm),99.4％(254nm).
实施例25.可参照化合物114、化合物23、化合物16的合成，分别制备得到以下化合物：
化合物115(甲酸盐)： 1H NMR(400MHz,DMSO-d 6,80℃)δ8.15(s,1H),7.35–7.22(m,5H),7.06–7.02(m,1H),6.30–6.27(m,1H),5.19(s,1H),3.14–3.07(m,2H),3.01–2.91(m,4H),2.88–2.77(m,3H),2.70–2.66(m,2H),2.62–2.58(m,2H),2.07–1.86(m,2H),1.80–1.71(m,2H).LC-MS:ESI m/z 409.3[M+H] +；C 23H 28N 4O 3计算值408.22.HPLC纯度:98.9％(214nm),99.3％(254nm).
化合物116(甲酸盐)： 1H NMR(400MHz,DMSO-d 6,80℃)δ8.10–8.07(m,1H),7.34(d,J＝7.2Hz,2H),7.26(dd,J＝8.0,7.2Hz,2H),7.23–7.17(m,1H),7.10–7.04(m,1H),6.72(brs,1H),6.32–6.25(dd,J＝18.8,7.2Hz,1H),5.08(t,J＝7.6Hz,1H),3.29–3.21(m,2H),2.97–2.85(m,1H),2.62–2.58(m,7H),2.46–2.36(m,3H),2.03–1.84(m,2H),1.77–1.73(m,4H).LC-MS:(ESI)m/z 423.2[M+H] +；C 24H 30N 4O 3计算值422.23.HPLC纯度:99.4％(254nm),99.7％(214nm).
化合物117(甲酸盐)： 1H NMR(400MHz,DMSO-d 6,80℃)δ7.42–7.26(m,5H),7.01(t,J＝6.4Hz,1H),6.28(t,J＝6.4Hz,1H),5.99(brs,1H),5.25(d,J＝6.8Hz,1H),3.57–3.44(m,4H),3.37–3.22(m,2H),3.12–3.03(m,1H),2.69(d,J＝7.6Hz,2H),2.60(t,J＝5.2Hz,2H),2.56–2.51(m,2H),2.16–2.04(m,2H),1.81–1.72(m,2H).LC-MS:(ESI)m/z 437.2[M+H] +；C 24H 28N 4O 4计算值436.21.HPLC纯度:98.9％(214nm),99.2％(254nm).
化合物121(甲酸盐)： 1H NMR(400MHz,DMSO-d 6)δ8.89–8.82(m,1H),7.76(brs,1H),7.47–7.25(m,5H),7.19(t,J＝8.0Hz,1H),6.37–6.34(m,1H),5.21(d,J＝6.4Hz,1H),3.35–2.93(m,9H),2.68–2.58(m,2H),2.49–2.44(m,2H),2.28–2.12(m,1H),2.05–1.91(m,1H),1.80–1.50(m,6H).LC-MS:ESI m/z 437.3[M+H] +；C 25H 32N 4O 3计算值436.25.HPLC纯度:100.0％(214nm),100.0％(254nm).
化合物122(甲酸盐)： 1H NMR(400MHz,DMSO-d 6)δ8.69–8.37(m,1H),7.47–6.95(m,6H),6.46–6.18(m,1H),5.36–5.15(m,1H),3.77–3.33(m,3H),3.32–2.99(m,4H),2.71–2.52(m,2H),2.49–2.41(m,2H),2.34–2.14(m,2H),2.09–1.89(m,2H),1.84–1.60(m,4H).LC-MS:ESI m/z 451.2[M+H] +；C 25H 30N 4O 4计算值450.23.HPLC纯度:97.4％(214nm),96.5％(254nm).
实施例26 化合物125的合成
Figure PCTCN2021078537-appb-000174
步骤1：3-((((R)-2-甲氧基-2-氧代-1-苯基乙基)氨基)甲基)吡咯烷-1-甲酸叔丁酯
Figure PCTCN2021078537-appb-000175
在N 2气氛下，向3-甲酰基-1-吡咯烷甲酸叔丁酯(5.0g，25mmol)的二氯乙烷(30mL)溶液中，加入(2R)-2-氨基-2-苯基乙酸甲酯(5.0g，30mmol)和冰醋酸(75mg，1.3mmol)。反应液在25℃下搅拌0.5小时后，将三乙酰氧基硼氢化钠(11g，50mmol)加入到反应液中，继续反应14小时。用水淬灭反应，并用DCM(50mL×2)萃取。合并的有机相用饱和NaHCO 3溶液(50mL)和盐水(50mL)洗涤，经硫酸钠干燥，过滤并浓缩，得到黄色油状物，将其通过硅胶柱色谱法纯化(石油醚：乙酸乙酯＝8：1-5：1)，得到标题化合物(7.0g，80％)，为黄色油状物。 1H NMR(400MHz,CDCl 3)δ7.38–7.29(m,5H),4.36(d,J＝2.8Hz,1H),3.70(s,3H),3.54–3.42(m,4H),3.12–2.87(m,1H),2.63–2.42(m,2H),2.36–2.26(m,1H),1.53-1.60(m,2H)1.45(s,9H).LC-MS:(ESI)m/z 349.2[M+H] +,C 19H 28N 2O 4计算值348.20.
步骤2：3-((((R)-2-甲氧基-2-氧代-1-苯基乙基)(甲基)氨基)甲基)吡咯烷-1-甲酸叔丁基
Figure PCTCN2021078537-appb-000176
向3-((((R)-2-甲氧基-2-氧代-1-苯基乙基)氨基)甲基)吡咯烷-1-甲酸叔丁酯(3.7g，11mmol)在DCE(40mL)中的溶液中加入多甲醛(3.0g，32mmol)和冰醋酸(32mg，0.10mmol)。将反应液在25℃搅拌0.5小时后，加入三乙酰氧基硼氢化钠(3.4g，16mmol)。将混合物再在50℃下搅拌14小时。加入水(50mL)将反应淬灭，用DCM(30mL×2)萃取。合并的有机相用NaHCO 3溶液(50mL)洗涤，然后用盐水(50mL)洗涤，经硫酸钠干燥，过滤并浓缩至干。通过硅胶柱色谱法纯化(石油醚：乙酸乙酯＝8：1-3：1)，得到标 题化合物(3.8g，52％)，为无色油状物。LC-MS:(ESI)m/z 363.3[M+H] +,C 20H 30N 2O 4计算值362.22.
步骤3：(2R)-2-(甲基(吡咯烷-3-基甲基)氨基)-2-苯基乙酸甲酯
Figure PCTCN2021078537-appb-000177
向3-(((((R)-2-甲氧基-2-氧代-1-苯基乙基)(甲基)氨基)甲基)吡咯烷-1-甲酸叔丁酯(3.8g，10mmol)的甲醇溶液中加入4N的盐酸/1，4-二氧六环溶液(15mL)。在50℃下反应3小时后，将反应物浓缩，得到呈白色固体的标题化合物的粗品(3.9g，粗品)。LC-MS:(ESI)m/z 263.2[M+H] +,C 15H 22N 2O 2计算值262.17.
步骤4-6：参照化合物114的步骤3-5合成制备得到化合物125(铵盐). 1H NMR(400MHz,DMSO-d 6,80℃)δ8.13(s,1H),7.37–7.28(m,5H),7.03–7.00(m,1H),6.29–6.27(m,1H),5.96(brs,1H),4.19–4.14(m,1H),3.43–3.28(m,2H),3.26–2.88(m,2H),2.68–2.54(m,5H),2.48–2.31(m,6H),2.20–2.18(m,3H),1.93–1.88(m,1H),1.79–1.74(m,2H),1.62–1.46(m,1H).LC-MS:(ESI)m/z 437.4[M+H] +；C 25H 32N 4O 3计算值436.25.HPLC Purity:99.7％(254nm),99.9％(214nm).
实施例27.参照化合物125的合成，制备得到以下化合物：
化合物126(甲酸盐)： 1H NMR(400MHz,DMSO-d 6)δ7.43(t,J＝6.0Hz,2H),7.37–7.30(m,3H),7.09–7.04(m,1H),6.72–6.46(m,1H),6.28(t,J＝7.5Hz,1H),4.28–4.26(m,1H),3.35–3.24(m,2H),2.95–2.88(m,4H),2.79–2.69(m,4H),2.67–2.60(m,3H),2.58–2.50(m,2H),2.10–1.90(m,1H),1.80–1.65(m,2H),1.65–1.51(m,1H).LC-MS:(ESI)m/z 395.3[M+H] +；C 23H 30N 4O 2计算值394.24.HPLC纯度:99.2％(214nm),99.1％(254nm).
化合物127(甲酸盐)： 1H NMR(400MHz,DMSO-d 6,80℃)δ7.40–7.38(m,2H),7.34–7.25(m,3H),7.06–7.02(m,1H),6.31–6.27(m,1H),5.98(brs,1H),4.24(s,1H),3.64–3.42(m,4H),3.44–3.01(m,5H),2.65–2.52(m,3H),2.43–2.31(m,1H),2.02–1.88(m,2H),1.79–1.75(m,2H),1.65–1.53(m,1H).LC-MS:ESI m/z 409.3[M+H] +；C 23H 28N 4O 3计算值408.22.HPLC纯度:99.9％(214nm),99.9％(254nm).
化合物128(甲酸盐)： 1H NMR(400MHz,DMSO-d 6,80℃)δ7.41-7.39(m,2H),7.34–7.27(m,3H),7.06(dd,J＝7.2,2.4Hz,1H),6.30–6.28(m,1H),4.27(s,1H),3.27–3.24(m,2H),3.11-2.97(m,3H),2.87-2.81(m,3H),2.66–2.57(m,4H),2.53–2.47(m,3H),2.05-2.00(m,1H),1.92–1.87(m,2H),1.79–1.73(m,2H),1.61–1.58(m,1H).LC-MS:(ESI)m/z 409.4[M+H] +；C 24H 32N 4O 2计算值408.25.HPLC Purity:99.2％(214nm),98.8％(254nm).
化合物129(甲酸盐): 1H NMR(400MHz,DMSO-d 6)δ7.40(d,J＝7.6Hz,1H),7.34–7.25(m,4H),7.01(d,J＝7.2Hz,1H),6.27(d,J＝7.2Hz,1H),4.26(s,1H),3.54–3.42(m,4H),3.2-2.99(m,2H),2.68-2.58(m,8H),2.31–2.29(m,1H),1.97-1.89(m,1H),1.78–1.72(m,2H),1.67–1.45(m,1H).LC-MS:(ESI)m/z 423.3[M+H] +；C 24H 30N 4O 3计算值422.23.
化合物130(甲酸盐)： 1H NMR(400MHz,DMSO-d 6,80℃)δ7.45–7.23(m,5H),7.10–7.04(m,1H),6.31–6.26(m,1H),4.25(s,1H),3.28–3.24(m,2H),2.77–2.75(m,2H),2.64–2.55(m, 7H),2.49–2.35(m,4H),2.00–1.93(m,1H),1.78–1.76(m,2H),1.62–1.50(m,5H).LC-MS:ESI m/z 423.3[M+H] +；C 25H 34N 4O 2计算值422.27.HPLC纯度:98.4％(254nm),97.6％(214nm).
化合物131(甲酸盐)： 1H NMR(400MHz,DMSO-d 6,80℃)δ7.41–7.40(m,2H),7.37–7.23(m,3H),7.05–7.02(m,1H),6.27–6.25(m,1H),6.22–6.04(m,1H),4.26(s,1H),3.52–3.32(m,3H),3.28–3.01(m,3H),2.65–2.51(m,4H),2.47–2.30(m,3H),2.21–2.20(m,2H),2.08–1.91(m,1H),1.87–1.74(m,4H),1.63–1.52(m,1H).LC-MS:ESI m/z 437.3[M+H] +；C 25H 32N 4O 3计算值436.25.HPLC纯度:99.4％(254nm),98.0％(214nm).
化合物134(甲酸盐)： 1H NMR(400MHz,DMSO-d 6)δ7.42–7.25(m,5H),7.10(dd,J＝19.2,7.2Hz,1H),6.32(t,J＝7.2Hz,1H),4.23–4.14(m,1H),3.30–3.22(m,2H),3.12–2.79(m,6H),2.76–2.72(m,2H),2.65–2.60(m,2H),2.52–2.50(m,1H),2.48–2.44(m,2H),2.19–2.22(m,3H),2.04–1.90(m,1H),1.82–1.72(m,2H),1.48–1.55(m,1H).LC-MS:(ESI)m/z 409.3[M+H] +；C 24H 32N 4O 2计算值408.25.HPLC纯度:99.2％(214nm),98.9％(254nm).
将化合物134混合物通过制备型HPLC(色谱柱：Kromasil-C18 100×21.2mm 5um，流动相：乙腈-水(0.1％甲酸)；梯度：10～20％)纯化，得到峰1化合物(18mg，28％)和峰2化合物(19.5mg，30％)。峰1化合物(18mg)通过SFC纯化(色谱柱：Daicel ChiralPak IG-3，3.0×150mm，3um，流动相：CO 2/甲醇(0.1％DEA)＝55/45，流速：1.5mL/min)以提供化合物134a和化合物134b，将峰2化合物定为化合物134c。
化合物134a： 1H NMR(400MHz,CD 3OD)δ7.48(d,J＝6.8Hz,2H),7.35–7.22(m,3H),7.15(d,J＝7.2Hz,1H),6.39(d,J＝7.2Hz,1H),3.98(s,1H),3.40–3.36(m,3H),3.28–3.23(m,2H),3.21–3.15(m,2H),3.02–2.98(m,1H),2.87(t,J＝6.8Hz,2H),2.70(t,J＝6.4Hz,2H),2.64–2.56(m,1H),2.47–2.43(m,1H),2.30(s,3H),2.19–2.08(m,1H),1.90–1.82(m,2H),1.80–1.70(m,1H),1.60–1.52(m,1H).LC-MS:ESI m/z 409.3[M+H] +；C 24H 32N 4O 2计算值408.25.HPLC纯度:98.3％(214nm),99.4％(254nm).
化合物134b： 1H NMR(400MHz,CD 3OD)δ7.52–7.50(m,2H),7.44–7.37(m,4H),6.54(d,J＝7.2Hz,1H),3.97(s,1H),3.56–3.41(m,6H),3.38–3.32(m,3H),3.09–3.03(m,2H),2.98–2.90(m,1H),2.72–2.68(m,2H),2.62(s,3H),2.35–2.26(m,1H),1.98–1.48(m,4H).LC-MS:ESI m/z 409.3[M+H] +；C 24H 32N 4O 2计算值408.25.HPLC纯度:96.4％(214nm),99.0％(254nm).
化合物134c： 1H NMR(400MHz,DMSO-d 6,80℃)δ7.43–7.31(m,6H),6.50(d,J＝7.2Hz,1H),4.37(s,1H),3.40–3.38(m,2H),3.36–3.30(m,6H),3.27–3.21(m,2H),2.98–2.91(m,2H),2.71–2.69(m,3H),2.27(s,3H),2.16–2.14(m,1H),1.85–1.77(m,2H),1.74–1.63(m,1H).
化合物136(甲酸盐)： 1H NMR(400MHz,DMSO-d 6,80℃)δ7.39–7.23(m,5H),7.01(t,J＝7.2Hz,1H),6.27–6.24(m,1H),6.08(brs,1H),4.14(s,0.5H),4.09(s,0.5H),3.24–3.23(m,2H),2.67–2.49(m,8H),2.45–2.21(m,5H),2.20(s,1.5H),2.15(s,1.5H),1.75–1.70(m,5H),1.38–1.24(m,1H).LC-MS:(ESI)m/z 423.3[M+H] +；C 25H 34N 4O 2计算值422.27.HPLC纯度:98.5％(214nm),97.9％(254nm).
化合物138(甲酸盐)：1H NMR(400MHz,DMSO-d 6,80℃)δ7.43–7.27(m,5H),7.11–7.05(m,1H),6.30(t,J＝7.3Hz,1H),4.24(s,0.55H),4.09(s,0.45H),3.28–3.25(m,2H),3.00–2.78(m,3H),2.79–2.66(m,3H),2.64–2.61(m,2H),2.60–2.51(m,3H),2.49–2.31(m,3H),2.21(s,1.65H),2.14(s,1.35H),1.98–1.91(m,1H),1.79–1.75(m,2H),1.66–1.47(m,4H).LC-MS: (ESI)m/z 437.3[M+H] +；C 26H 36N 4O 2计算值436.28.HPLC纯度:97.9％(254nm),98.0％(214nm).
化合物139(甲酸盐)： 1H NMR(400MHz,DMSO-d 6,80℃)δ7.39–7.27(m,5H),7.07(t,J＝7.2Hz,1H),6.55(brs,1H),6.29–6.27(m,1H),4.21–4.17(m,1H),3.48–3.41(m,1H),3.37–3.24(m,3H),3.21–3.19(m,1H),3.10–3.02(m,1H),2.63–2.60(m,2H),2.49–2.34(m,5H),2.25–2.13(m,5H),1.94–1.72(m,5H),1.62–1.47(m,1H).LC-MS:(ESI)m/z 437.3[M+H] +；C 26H 34N 4O 3计算值451.3.HPLC纯度:99.4％(254nm),99.9％(214nm).
实施例28.化合物140的合成
Figure PCTCN2021078537-appb-000178
步骤1：(R)-2-(甲氨基)-2-苯乙酸甲酯
Figure PCTCN2021078537-appb-000179
在25℃下向(R)-(甲氨基)苯乙酸(5.0g，30mmol)的甲醇(100mL)溶液中加入氯化亚砜(5mL)，将反应混合物在75℃下搅拌3小时。浓缩混合物，得到(R)-2-(甲基氨基)-2-苯基乙酸甲酯(5.5g，90％)，为白色固体。 1H NMR(400MHz,CDCl 3)δ:7.64(d,J＝4.4Hz,2H),7.50–7.40(m,3H),4.97(d,J＝4.4Hz,1H),3.79(s,3H),2.63(t,J＝4.7Hz,3H).
步骤2：3-((((R)-2-甲氧基-2-氧代-1-苯乙基)(甲基)氨基甲酰基)吡咯烷-1-甲酸叔丁酯
Figure PCTCN2021078537-appb-000180
在N 2气氛下，将1-羟基苯并三唑(0.49g，3.6mmol)和EDCI(0.66g，3.6mmol)加入到1-(叔丁氧羰基)吡咯烷-3-羧酸(0.50g，2.3mmol)，(R)-2-(甲氨基)-2-苯乙酸甲酯(0.50g，2.7mmol)和三乙胺(0.47g，4.6mmol)的混合液中。该反应液在25℃反应18小时后，用水(50mL)淬灭，并用二氯甲烷(30mL×2)萃取。合并的有机相用HCl(0.5N，10mL)洗涤，然后用盐水(50mL)洗涤，经硫酸钠干燥并浓缩，得到标题化合物(0.85g，粗品)，为淡黄色油状物，其无需进一步纯化直接用于下一步。 1H NMR(400MHz,CDCl 3)δ7.31(d,J ＝6.8Hz,3H),7.17–7.09(m,2H),6.36(d,J＝5.6Hz,1H),5.23(s,1H),3.73(d,J＝14.0Hz,3H),3.58–3.13(m,5H),2.83–2.72(m,3H),2.19–1.89(m,3H),1.39(d,J＝2.4Hz,9H).LC-MS:(ESI)m/z 399.3[M+Na] +.C 20H 28N 2O 5计算值376.20.
步骤3：(2R)-2-(N-甲基-1-吡咯烷基-3-甲酰胺基)-2-苯乙酸甲酯
Figure PCTCN2021078537-appb-000181
在N 2气氛下，向3-((((R)-2-甲氧基-2-氧代-1-苯乙基)(甲基)氨基甲酰基)吡咯烷-1-甲酸叔丁酯(3.6g，10mmol)的甲醇(5mL)溶液中，加入4N的盐酸/1，4-二氧六环溶液(25mL，0.1mol)，并在25℃下搅拌3小时后，浓缩混合物，得到标题化合物(3.9g，粗品)，为红色油状物，其无需进一步纯化直接用于下一步。LC-MS:(ESI)m/z 277.2[M+H] +,C 15H 20N 2O 3计算值276.15.
步骤4-6：参照化合物114步骤3-5合成制备得到化合物140(甲酸盐). 1H NMR(400MHz,DMSO-d 6)δ7.40–7.31(m,5H),7.03(d,J＝6.8Hz,1H),6.31(d,J＝7.2Hz,1H),6.10–6.00(m,1H),3.63–3.45(m,6H),3.30–3.24(t,J＝4.0Hz,3H),2.84(s,2H),2.73–2.71(m,2H),2.63(t,J＝6.0Hz,2H),2.55–2.52(m,2H),2.08–1.95(m,2H),1.84–1.74(m,2H).LC-MS:(ESI)m/z 451.3[M+H] +；C 25H 30N 4O 4计算值450.23.HPLC纯度:98.6％(214nm),98.6％(254nm).
实施例29.参照化合物140的合成，制备得到以下化合物：
化合物141(甲酸盐)： 1H NMR(400MHz,DMSO-d 6,80℃)δ7.43–7.17(m,5H),7.03(t,J＝7.6Hz,1H),6.32–6.29(m,1H),6.06(brs,1H),3.32–3.24(m,4H),2.97–2.93(m,1H),2.84–2.56(m,12H),1.99–1.93(m,2H),1.77–1.73(m,2H).LC-MS:ESI m/z 423.3[M+H] +；C 24H 30N 4O 3计算值422.23.HPLC纯度:99.3％(214nm),99.8％(254nm).
化合物142(铵盐)： 1H NMR(400MHz,DMSO-d 6,80℃)δ7.39–7.20(m,5H),7.12–7.00(m,1H),6.37–6.26(m,1H),6.12–5.95(m,1H),3.66–3.57(m,2H),3.51–3.39(m,4H),3.33–3.19(m,5H),2.83–2.77(m,2H),2.64–2.58(m,2H),2.09–1.93(m,2H),1.80–1.70(m,2H).LC-MS:ESI m/z 437.3[M+H] +；C 24H 28N 4O 4计算值436.21.HPLC纯度:98.2％(214nm),97.9％(254nm).
化合物143(甲酸盐)： 1H NMR(400MHz,DMSO-d 6)δ7.33–7.27(m,5H),7.02–7.00(m,1H),6.26(t,J＝6.4Hz,1H),6.07(brs,1H),3.25(s,3H),2.75–2.55(m,8H),2.48–2.41(m,6H),2.08–1.86(m,2H),1.81–1.68(m,4H).LC-MS:(ESI)m/z 437.3[M+H] +；C 25H 32N 4O 3计算值436.25.HPLC纯度:95.3％(214nm),93.9％(254nm).
化合物144(甲酸盐)： 1H NMR(400MHz,DMSO-d 6)δ7.43–7.20(m,5H),7.13–6.99(m,1H),6.32–6.23(m,1H),6.08(brs,1H),3.39–3.22(m,3H),3.00–2.54(m,10H),2.49–2.42(m,4H),2.16–1.88(m,2H),1.81–1.42(m,6H).LC-MS:ESI m/z 451.3[M+H] +；C 26H 34N 4O 3计算值450.26.HPLC纯度:99.8％(214nm),100.0％(254nm).
化合物145(甲酸盐)： 1H NMR(400MHz,DMSO-d 6,80℃)δ7.43–6.99(m,6H),6.39–6.23(m,1H),6.13–5.97(m,1H),3.80–3.15(m,8H),2.90–2.69(m,3H),2.67–2.58(m,2H),2.48–2.44(m,2H),2.31–2.16(m,2H),2.14–1.94(m,2H),1.90–1.68(m,4H).LC-MS:ESI m/z 465.3[M+H] +；C 26H 32N 4O 4计算值464.24.HPLC纯度:99.4％(214nm),98.4％(254nm).
实施例30.化合物146的合成
Figure PCTCN2021078537-appb-000182
步骤1：2-(2-(三氟甲基)苯基)乙酸甲酯
Figure PCTCN2021078537-appb-000183
向2-(3-(三氟甲基)苯基)乙酸(2.0g，9.8mmol)的甲醇(20mL)溶液中加入浓H 2SO 4(2.0mL)，并将所得混合物在80℃下搅拌18小时。浓缩至干后，将残余物用水(20mL)稀释，并用乙酸乙酯(20mL×2)萃取。合并的有机相用饱和NaHCO 3(20mL)和盐水洗涤，用硫酸钠干燥，过滤并浓缩，得到标题化合物的粗产物(2.0g，95％)，为无色油状物。 1H NMR(400MHz,DMSO-d 6)δ7.72(d,J＝7.8Hz,1H),7.65(d,J＝7.3Hz,1H),7.52(d,J＝8.9Hz,2H),3.89(d,J＝1.2Hz,2H),3.62(s,3H).LC-MS:ESI m/z无离子峰[M+H] +；C 10H 9F 3O 2计算值218.06.
步骤2：2-溴-2-(2-(三氟甲基)苯基)乙酸甲酯
Figure PCTCN2021078537-appb-000184
在N 2气氛下，向2-(2-(三氟甲基)苯基)乙酸甲酯(2.0g，9.2mmol)在CCl 4(20mL)的溶液中添加AIBN(150mg，0.92mmol)和NBS(1.8g，10mmol)。在80℃下搅拌14小时后，将反应用饱和Na 2S 2O 3(10mL)和NaHCO 3(10mL)淬灭，水层用二氯甲烷(20mL)萃取。合并的有机相用盐水洗涤，经硫酸钠干燥，过滤并浓缩至干。残余物通过快速柱色谱法纯化(0-10％乙酸乙酯的石油醚溶液)，得到标题化合物(2.4g，89％)，为浅黄色油状物。 1H NMR(400MHz,DMSO-d 6)δ7.85–7.76(m,3H),7.62(t,J＝7.5Hz,1H),5.92(s,1H),3.74(s,3H).LC-MS:ESI m/z no ionic peak[M+H] +；C 10H 8BrF 3O 2计算值295.97.
步骤3：(3S)-3-((2-甲氧基-2-氧-1-(2-(三氟甲基)苯基)乙基)氨基)吡咯烷-1-羧酸叔丁酯
Figure PCTCN2021078537-appb-000185
将2-溴-2-(2-(三氟甲基)苯基)乙酸甲酯(1.4g，4.7mmol)，(S)-3-氨基吡咯烷-1-羧酸叔丁酯(1.0g，5.6mmol)和K 2CO 3(1.9g，14mmol)在乙腈(20mL)中的混合物在25℃下搅拌16小时。用水(20mL)将反应淬灭，并用乙酸乙酯(15mL×2)萃取。合并的有机相用盐水洗涤，经硫酸钠干燥，过滤并浓缩至干。残余物通过快速柱色谱法纯化(0-40％乙酸乙酯的石油醚溶液)，得到标题化合物(744mg，41％)，为黄色油状物。LC-MS:ESI m/z 403.1[M+H] +；C 19H 25F 3N 2O 4计算值402.18.
步骤4-7：参照化合物5的步骤2-5合成方法，制备得到白色固体的化合物146(甲酸盐)(29.8mg，13％)。 1H NMR(400MHz,CD 3OD)δ7.96(d,J＝7.9Hz,1H),7.70(d,J＝7.9Hz,1H),7.59(t,J＝7.6Hz,1H),7.46(t,J＝7.6Hz,1H),7.34(d,J＝7.3Hz,1H),6.44(d,J＝7.3Hz,1H),4.63–4.61(m,1H),3.97–3.87(m,1H),3.43–3.37(m,2H),3.36–3.32(m,2H),3.29–3.21(m,2H),3.13–3.00(m,2H),2.78–2.65(m,4H),2.31–2.19(m,3H),2.19–2.10(m,2H),2.06–1.94(m,2H),1.93–1.85(m,2H).LC-MS:ESI m/z 477.3[M+H] +；C 25H 31F 3N 4O 2计算值476.24.
实施例31.参照化合物146的合成方法，制备得到以下化合物：
Figure PCTCN2021078537-appb-000186
化合物147(甲酸盐)： 1H NMR(400MHz,CD 3OD)δ8.01–7.94(m,1H),7.67(d,J＝7.8Hz,1H),7.59–7.57(m,1H),7.43(t,J＝7.4Hz,1H),7.37–7.34(m,1H),6.45(t,J＝6.7Hz,1H),4.60(s,1H),4.00–3.83(m,1H),3.48–3.38(m,3H),3.36–3.31(m,2H),3.29–3.22(m,1H),3.15–3.04(m,2H),2.74(t,J＝6.1Hz,2H),2.58–2.55(m,2H),2.31–2.19(m,3H),2.17–2.06(m,2H),1.94–1.86(m,2H),1.77–1.62(m,4H),1.48–1.37(m,2H).LC-MS:ESI m/z 505.26[M+H] +；C 27H 35F 3N 4O 2计算值504.27.
实施例32.化合物149的合成
Figure PCTCN2021078537-appb-000187
步骤1：2-溴-2-(3-氯苯基)乙酸甲酯
Figure PCTCN2021078537-appb-000188
在N 2气氛下，向2-(3-氯苯基)乙酸甲酯(1.0g，5.41mmol)的CCl 4(20mL)溶液中加入NBS(1.06g，5.95mmol)和AIBN(89mg，0.54mmol)。将该混合物回流16小时。将混合物过滤并将滤液真空浓缩。将残余物通过快速柱色谱法纯化(0-15％乙酸乙酯的石油醚溶液)，得到标题化合物(1.0g，70％)，为无色油状物。 1H NMR(400MHz,CDCl 3)δ7.48(t,J＝1.7Hz,1H),7.36–7.33(m,1H),7.25–7.22(m,2H),5.22(s,1H),3.72(s,3H).
步骤2：7-(5-((3S)-3-((1-(3-氯苯基)-2-甲氧基-2-氧乙基)(甲基)氨基)吡咯烷-1-基)戊基)-3,4-二氢-1,8-萘啶-1(2H)-羧酸叔丁酯
Figure PCTCN2021078537-appb-000189
向2-溴-2-(3-氯苯基)乙酸甲酯(179mg，0.68mmol)在乙腈(8mL)中的溶液中加入(S)-7-(5-(3-(甲基氨基))吡咯烷-1-基)戊基)-3,4-二氢-1,8-萘啶-1(2H)-羧酸叔丁酯(250mg，0.62mmol)和K 2CO 3(171mg，1.24mmol)。将混合物在室温搅拌16小时。真空除去溶剂。通过在SiO 2(0-3％甲醇的二氯甲烷溶液)中的快速柱色谱法纯化残余物，得到标题化合物(120mg，33％)，为黄色油状物。LC-MS:ESI m/z:585.2.[M+H] +；C 32H 45ClN 4O4 require 584.31.
步骤3：2-(3-氯苯基)-2-(甲基((S)-1-(5-(5,6,7,8-四氢-1,8-萘啶-2-基)戊基)吡咯烷酮-3-基)氨基)乙酸
Figure PCTCN2021078537-appb-000190
在室温下将7-(5-((3S)-3-((1-(3-氯苯基)-2-甲氧基-2-氧乙基)(甲基)氨基)吡咯烷-1-基)戊基)-3,4-二氢-1,8-萘啶-1(2H)-羧酸叔丁酯(120mg，0.21mmol)的盐酸/1，4-二氧六环(4N，4mL)溶液搅拌3小时。真空浓缩溶剂。将残余物溶于甲醇(4mL)和H 2O(1mL)中，然后加入LiOH(25mg，1.05mmol)。将混合物在40℃下搅拌3小时。通过1N的甲酸将混合物调节至pH≈7。真空浓缩溶剂。残余物通过制备型HPLC在以下条件下纯化[柱：Kromasil Prep C18，30×150mm；流动相：1-40％乙腈-水溶液(含0.1％甲酸)；时间：15分钟]，得到化合物149(33.2mg，33％)，为无色油状物。 1H NMR(400MHz,CD 3OD)δ7.53(d,J＝9.7Hz,1H),7.46(dd,J＝7.3,3.7Hz,1H),7.40(dd,J＝7.0,1.5Hz,1H),7.36–7.23(m,2H),6.52(dd,J＝7.3,3.2Hz,1H),4.37(s,0.5H),4.28(s,0.5H),3.86–3.76(m,1H),3.48-3.33(m,5H), 3.14–3.06(m,3H),2.78(t,J＝5.5Hz,2H),2.69–2.59(m,2H),2.38(s,1.5H),2.26(s,1.5H),2.20–2.10(m,2H),1.96–1.88(m,2H),1.78–1.69(m,4H),1.52–1.43(m,2H).LC-MS:ESI m/z 471.3,473.2[M+H,Cl] +；C 26H 35ClN 4O 2计算值470.24.
实施例33.参照化合物149的合成方法，制备得到以下化合物：
化合物150(甲酸盐)： 1H NMR(400MHz,CD 3OD)δ7.67–7.60(m,1H),7.49–7.42(m,1H),7.40–7.25(m,3H),6.50–6.47(m,1H),4.95(s,0.5H),4.94(s,0.5H),3.97–3.87(m,1H),3.57–3.33(m,4H),3.09–2.88(m,4H),2.83–2.65(m,4H),2.47(s,1.5H),2.35(s,1.5H),2.29–2.19(m,2H),2.04–1.83(m,4H).LC-MS:ESI m/z 443.2,445.1[M+H,Cl] +；C 24H 31ClN 4O 2计算值442.21.
化合物151(甲酸盐)： 1H NMR(400MHz,CD 3OD)δ7.55(d,J＝7.0Hz,1H),7.43–7.34(m,4H),6.50(dd,J＝7.3,1.8Hz,1H),4.43(s,0.5H),4.34(s,0.5H),3.88–3.77(m,1H),3.46–3.32(m,4H),3.1–2.84(m,4H),2.78–2.69(m,4H),2.44(s,1.5H),2.32(s,1.5H),2.25–2.08(m,2H),2.04–1.86(m,4H).LC-MS:ESI m/z 443.2,445.3[M+H,Cl] +；C 24H 31ClN 4O 2计算值442.21.
化合物152(甲酸盐)： 1H NMR(400MHz,CD 3OD)δ7.48(d,J＝8.4Hz,2H),7.43–7.27(m,3H),6.48(d,J＝7.3Hz,1H),4.41(d,J＝37.4Hz,1H),3.88–3.74(m,1H),3.55–3.32(m,4H),3.06–2.82(m,4H),2.81–2.66(m,4H),2.39(d,J＝47.3Hz,3H),2.23–2.09(m,2H),2.01–1.79(m,4H).LC-MS:ESI m/z 443.2,445.3[M+H,Cl] +；C 24H 31ClN 4O 2计算值442.21.
化合物154(甲酸盐)： 1H NMR(400MHz,CD 3OD)δ7.86–7.80(m,1H),7.77–7.72(m,1H),7.66–7.61(m,1H),7.59–7.54(m,1H),7.41–7.35(m,1H),6.50(dd,J＝7.3,1.8Hz,1H),4.55(s,0.5H),4.42(s,0.5H),3.84–3.77(m,1H),3.61–3.48(m,1H),3.45–3.36(m,3H),3.16–3.07(m,1H),3.06–2.96(m,3H),2.81–2.69(m,4H),2.43(s,1.5H),2.27(s,1.5H),2.24–2.11(m,2H),2.06–1.94(m,2H),1.93–1.87(m,2H).LC-MS:ESI m/z 477.2[M+H] +；C 25H 31F 3N 4O 2计算值476.24.
化合物155(甲酸盐)： 1H NMR(400MHz,CD 3OD)δ7.73–7.56(m,4H),7.29–7.22(m,1H),6.44(d,J＝7.2Hz,1H),4.45–4.40(m,0.5H),4.35(s,0.5H),3.82–3.72(m,1H),3.51–3.41(m,1H),3.40–3.32(m,3H),3.24–3.15(m,1H),3.13–3.01(m,3H),2.77–2.63(m,4H),2.40(s,1.5H),2.28(s,1.5H),2.22–2.12(m,2H),2.03–1.93(m,2H),1.90–1.82(m,2H).LC-MS:ESI m/z 477.3[M+H] +,C 25H 31F 3N 4O 2.计算值476.24.
化合物156(甲酸盐)： 1H NMR(400MHz,CD 3OD)7.53-7.40(m,3H),7.14–7.10(m,1H),7.05–7.02(m,1H),6.60–6.57(m,1H),5.40(s,0.4H),5.16(s,0.6H),3.97–3.95(m,1H),3.90-3.89(m,3H),3.50–3.34(m,3H),3.27–3.21(m,1H),3.06–2.86(m,1H),2.82–2.52(m,6H),2.52–2.46(m,3H),2.43–2.32(m,1H),2.32–2.24(m,2H),1.95–1.76(m,4H).LC-MS:ESI m/z 439.3[M+H] +,C 25H 34N 4O 3计算值438.26.
化合物157(甲酸盐)： 1H NMR(400MHz,CD 3OD)δ7.48–7.46(m,1H),7.34–7.30(m,1H),7.10–6.94(m,3H),6.55–6.53(m,1H),4.57(s,0.5H),4.47(s,0.5H),3.91–3.84(m,1H),3.80–3.79(m,3H),3.45–3.33(m,4H),3.26–3.21(m,2H),2.83–2.73(m,6H),2.55–2.44(m,3H),2.25–2.13(m,2H),1.96–1.85(m,4H).LC-MS:ESI m/z 439.3[M+H] +,C 25H 34N 4O 3计算值438.26.
化合物158(甲酸盐)： 1H NMR(400MHz,CD 3OD)δ7.45–7.43(m,3H),6.98–6.95(m,2H),6.54–6.51(m,1H),4.61(s,0.5H),4.50(s,0.5H),4.00–3.81(m,1H),3.81–3.80(m,3H),3.45–3.33(m,3H),3.19–3.13(m,1H),2.92–2.78(m,1H),2.77–2.75(m,2H),2.73–2.60(m,4H), 2.58–2.56(m,1.5H),2.52–2.47(m,1H),2.46–2.43(m,1.5H),2.21–2.11(m,2H),1.92–1.80(m,4H).LC-MS:ESI m/z 439.3[M+H] +；C 25H 34N 4O 3计算值438.26.
通过制备HPLC在以下条件下纯化化合物158[柱：Kromasil 100-5-C18，30×150mm；流动相：在14分钟内在1-100％乙腈的水溶液(含0.1％的甲酸)]，得到化合物158a和化合物158b，为白色固体。
化合物158a： 1H NMR(400MHz,CD 3OD)δ7.35–7.33(m,2H),7.20–7.18(m,1H),6.85–6.83(m,2H),6.35–6.33(m,1H),4.64(s,1H),3.81–3.80(m,1H),3.69(s,3H),3.30–3.28(m,2H),3.25(s,1H),3.11–3.06(m,1H),2.64–2.61(m,6H),2.54–2.50(m,2H),2.32–2.28(m,3H),2.07–2.05(m,2H),1.81–1.78(m,4H).LC-MS:ESI m/z 439.2[M+H] +,C 25H 34N 4O 3.计算值438.26.
化合物158b： 1H NMR(400MHz,CD 3OD)δ7.33–7.32(m,2H),7.07–7.02(m,1H),6.82–6.81(m,2H),6.29–6.27(m,1H),4.99(s,0.5H),4.63(s,0.5H),4.27–4.13(m,1H),3.69(s,3H),3.62–3.56(m,1H),3.25(s,2H),3.13–2.99(m,1H),2.71–2.57(m,6H),2.53–2.48(m,2H),2.45–2.33(m,3H),2.05–1.92(m,2H),1.84–1.74(m,4H).LC-MS:ESI m/z 439.1[M+H] +,C 25H 34N 4O 3计算值438.26.
化合物159(甲酸盐)： 1H NMR(400MHz,CD 3OD)δ7.61(dd,J＝7.3,2.2Hz,1H),7.51–7.38(m,2H),7.31–7.27(m,2H),6.51(dd,J＝7.3,2.4Hz,1H),4.86(s,0.5H),4.78(s,0.5H),3.96–3.86(m,1H),3.54–3.34(m,5H),3.23–3.09(m,3H),2.77(t,J＝6.1Hz,2H),2.68–2.58(m,2H),2.44(s,1.5H),2.31(s,1.5H),2.21–2.11(m,2H),1.91–1.89(m,2H),1.77–1.69(m,4H),1.50–1.44(m,2H).LC-MS:ESI m/z 471.3,473.1[M+H,Cl] +；C 26H 35ClN 4O 2计算值470.24.
将非对映异构体混合物化合物159(30mg，0.063mmol)在制备SFC手性分离(Chirex S-VAL，R-NEA，250×4.6mm)，用40％甲醇的二氧化碳溶液(0.05％DEA)洗脱，以3mL/min的速度浓缩得到化合物159a和化合物159b.
化合物159a： 1HNMR(400MHz,CD 3OD)δ7.53(dd,J＝7.4,1.7Hz,1H),7.32–7.28(m,2H),7.23–7.16(m,2H),6.38(d,J＝7.3Hz,1H),4.67(s,1H),3.87–3.76(m,1H),3.48–3.23(m,5H),3.10–2.93(m,3H),2.65(t,J＝6.2Hz,2H),2.52(t,J＝7.1Hz,2H),2.34(s,3H),2.16–1.98(m,2H),1.86–1.76(m,2H),1.71–1.51(m,4H),1.44–1.30(m,2H).LC-MS:ESI m/z:471.2,473.3[M+H,Cl] +；C 26H 35ClN 4O 2计算值470.24.采用分析型手性SFC(Chirex S-VAL和R-NEA(250×4.6mm)柱子)得到保留时间：8.709min，ee值：100.0％.
化合物159b： 1H NMR(400MHz,CD 3OD)δ7.59–7.49(m,1H),7.36–7.26(m,2H),7.25–7.13(m,2H),6.37(d,J＝7.3Hz,1H),4.76(s,1H),3.89–3.75(m,1H),3.33–3.23(m,5H),3.17–3.10(m,1H),3.04–2.93(m,2H),2.67–2.64(m,2H),2.57–2.46(m,2H),2.21(s,3H),2.14–2.09(m,2H),1.83–1.77(m,2H),1.66–1.56(m,4H),1.41–1.28(m,2H).LC-MS:ESI m/z:471.2,473.3[M+H,Cl] +；C 26H 35ClN 4O 2计算值470.24.采用分析型手性SFC(Chirex S-VAL和R-NEA(250×4.6mm)柱子)得到保留时间：9.686min，ee值：100.0％.
化合物160(甲酸盐)： 1H NMR(400MHz,CD 3OD)δ7.47(dd,J＝8.4,4.3Hz,2H),7.40(t,J＝7.6Hz,1H),7.37–7.29(m,2H),6.49(dd,J＝7.3,4.9Hz,1H),4.37(s,0.5H),4.28(s,0.5H),3.85–3.73(m,1H),3.50–3.31(m,5H),3.14–2.96(m,3H),2.78–2.72(m,2H),2.66–2.56(m,2H),2.39(s,1.5H),2.28(s,1.5H),2.21–2.08(m,2H),1.95–1.86(m,2H),1.77–1.60(m,4H),1.51–1.42(m 2H).LC-MS:ESI m/z 471.3,473.2[M+H,Cl] +；C 26H 35ClN 4O 2计算值470.24.
化合物161(甲酸盐)： 1H NMR(400MHz,CD 3OD)δ7.88–7.79(m,1H),7.77–7.70(m,1H), 7.65–7.49(m,2H),7.41(t,J＝7.8Hz,1H),6.49(dd,J＝7.2,5.9Hz,1H),4.42(s,0.5H),4.33(s,0.5H),3.89–3.76(m,1H),3.52–3.41(m,3H),3.40–3.32(m,2H),3.27–3.16(m,1H),3.17–2.95(m,2H),2.82–2.71(m,2H),2.68–2.57(m,2H),2.37(s,1.5H),2.23(s,1.5H),2.21–2.01(m,2H),1.95–1.86(m,2H),1.81–1.63(m,4H),1.53–1.39(m,2H).LC-MS:ESI m/z 505.37[M+H] +；C 27H 35F 3N 4O 2计算值504.27.
化合物162(甲酸盐)： 1H NMR(400MHz,CD 3OD)δ7.53–7.52(m,1H),7.42–7.38(m,2H),7.10–7.07(m,1H),7.04–6.97(m,1H),6.57–6.55(m,1H),5.12–5.07(m,1H),3.96–3.90(m,1H),3.88–3.87(m,3H),3.47–3.44(m,2H),3.43–3.32(m,2H),3.13–2.83(m,4H),2.81–2.77(m,2H),2.70–2.65(m,2H),2.50–2.47(m,3H),2.26–2.17(m,2H),1.95–1.89(m,2H),1.75–1.69(m,4H),1.48–1.44(m,2H).LC-MS:ESI m/z 467.30[M+H] +,C 27H 38N 4O 3计算值466.29.
化合物166(甲酸盐)： 1H NMR(400MHz,CD 3OD)δ7.65(dt,J＝8.2,5.7Hz,4H),7.44(dd,J＝7.3,3.7Hz,1H),6.48(dd,J＝7.3,5.0Hz,1H),4.37(s,0.5H),4.32(s,0.5H),3.82(m,1H),3.50–3.40(m,3H),3.41–3.32(m,2H),3.28–3.18(m,1H),3.16–3.05(m,2H),2.77(t,J＝5.9Hz,2H),2.66–2.57(m,2H),2.35(s,1.5H),2.24(s,1.5H),2.21–2.05(m,2H),1.96–1.86(m,2H),1.78–1.62(m,4H),1.49–1.35(m,2H).LC-MS:ESI m/z 505.2[M+H] +；C 27H 35F 3N 4O 2计算值504.27.
在以下条件下，通过制备型手性SFC分离化合物166(20mg，0.04mmol)[柱：Daicel ChiralPak IG(250×30mm，10um)；流动相：0.1％NH 3.H 2O的甲醇溶液；梯度：60％甲醇，6.5min；流速：70g/min]，分别得到黄色固体状的化合物166a和化合物166b。
化合物166a： 1H NMR(400MHz,CD 3OD)δ7.61–7.48(m,5H),6.53(d,J＝7.3Hz,1H),4.57(s,1H),3.79–3.65(m,1H),3.49–3.36(m,3H),3.35–3.23(m,3H),3.16–3.05(m,2H),2.75–2.69(m,2H),2.67–2.59(m,2H),2.26(s,3H),2.20–2.07(m,2H),1.89–1.81(m,2H),1.74–1.60(m,4H),1.42–1.31(m,2H).LC-MS:ESI m/z 505.2[M+H] +；C 27H 35F 3N 4O 2计算值504.27.采用分析型手性SFC(ChiralPak IG(250×30mm)柱子)得到保留时间：1.347min，ee值：96.9％.
化合物166b： 1H NMR(400MHz,CD 3OD)δ7.62–7.54(m,4H),7.49(d,J＝7.4Hz,1H),6.53(d,J＝7.4Hz,1H),4.57(s,1H),3.64(s,2H),3.46–3.34(m,3H),3.25(s,1H),3.15–3.03(m,3H),2.72(t,J＝6.1Hz,2H),2.67–2.60(m,2H),2.33(s,3H),2.11–1.95(m,2H),1.89–1.82(m,2H),1.73–1.61(m,4H),1.44–1.31(m,2H).LC-MS:ESI m/z 505.2[M+H] +；C 27H 35F 3N 4O 2计算值504.27.采用分析型手性SFC(ChiralPak IG(250×30mm)柱子)得到保留时间：2.256min，ee值：99.1％.
化合物167(甲酸盐)： 1H NMR(400MHz,CD 3OD)δ7.44–7.41(m,1H),7.29–7.25(m,1H),7.10–7.05(m,2H),6.90–6.88(m,1H),6.50–6.47(m,1H),4.37(s,0.5H),4.30(s,0.5H),3.93–3.81(m,1H),3.78(s,3H),3.48–3.41(m,2H),3.26–3.23(m,2H),3.13–2.97(m,2H),2.92–2.86(m,2H),2.76–2.75(m,2H),2.63–2.57(m,2H),2.45(s,1.5H),2.35(s,1.5H),2.18–2.09(m,2H),1.94–1.87(m,2H),1.70–1.68(m,4H),1.45–1.42(m,2H).LC-MS:ESI m/z 467.40[M+H] +,C 27H 38N 4O 3计算值466.29.
化合物168(甲酸盐)： 1H NMR(400MHz,CD 3OD)δ7.49–7.47(m,1H),7.43–7.39(m,2H),6.97–6.90(m,2H),6.55–6.53(m,1H),4.58(s,0.6H),4.43(s,0.4H),3.99–3.80(m,1H),3.80–3.79(m,3H),3.46–3.42(m,2H),3.26–3.13(m,1H),2.95–2.87(m,1H),2.83–2.76(m,5H),2.71–2.54(m,3H),2.49(s,1.5H),2.40(s,1.5H),2.19–2.04(m,2H),1.93–1.90(m,2H),1.70– 1.64(m,4H),1.51–1.37(m,2H).LC-MS:ESI m/z 467.30[M+H] +,C 27H 38N 4O 3.计算值466.29.
化合物172(甲酸盐)： 1H NMR(400MHz,CD 3OD)δ7.50–7.34(m,3H),7.11–7.07(m,1H),6.98(t,J＝7.5Hz,1H),6.52(d,J＝7.3Hz,1H),5.17(s,0.5H),5.07(s,0.5H),4.78–4.71(m,1H),4.03–3.92(m,1H),3.45–3.40(m,2H),3.30–3.15(m,2H),2.84–2.57(m,8H),2.52(s,1.5H),2.49(s,1.5H),2.29–2.14(m,2H),1.93–1.87(m,2H),1.76–1.45(m,6H),1.42–1.34(m,6H).LC-MS:ESI m/z 495.4[M+H] +；C 29H 42N 4O 3计算值494.33.
化合物173(甲酸盐)： 1H NMR(400MHz,CD 3OD)δ7.47–7.41(m,1H),7.24(t,J＝7.9Hz,1H),7.12–6.96(m,2H),6.87–6.84(m,1H),6.50–6.47(m,1H),4.59(m,1H),4.28(s,0.5H),4.26(s,0.5H),3.96–3.77(m,1H),3.46–3.40(m,2H),3.30–3.09(m,3H),3.08–2.86(m,3H),2.78–2.73(m,2H),2.61(t,J＝7.5Hz,2H),2.43(s,1.5H),2.34(s,1.5H),2.21–2.06(m,2H),1.94–1.87(m,2H),1.74–1.59(m,4H),1.46–1.36(m,2H),1.29(d,J＝5.9Hz,6H).LC-MS:ESI m/z 495.4[M+H] +；C 29H 42N 4O 3计算值494.33.
化合物177(甲酸盐)： 1H NMR(400MHz,CD 3OD)δ7.47(d,J＝7.3Hz,1H),7.39(d,J＝8.6Hz,2H),6.90(d,J＝8.6Hz,2H),6.51(d,J＝7.2Hz,1H),4.64–4.55(m,1H),4.44(s,0.5H),4.38(s,0.5H),3.96–3.77(m,1H),3.47–3.40(m,2H),3.29–3.18(m,2H),3.06–2.97(m,1H),2.86(t,J＝7.4Hz,3H),2.78–2.74(m,2H),2.64(t,J＝7.7Hz,2H),2.49(s,1.5H),2.40(s,1.5H),2.22–2.07(m,2H),1.95–1.87(m,2H),1.75–1.59(m,4H),1.48–1.38(m,2H),1.29(d,J＝6.0Hz,6H).LC-MS:ESI m/z 495.4[M+H] +；C 29H 42N 4O 3计算值494.33.
在以下条件下，通过制备型手性SFC分离差向异构体的混合物化合物177(15.8mg，0.032mmol)[色谱柱：Daicel ChiralPak IG(250×30mm，10um)；流动相：60％甲醇的二氧化碳溶液(含0.1％NH 3.H 2O)；流速：70g/min]，分别得到化合物177a和化合物177b，为黄色固体。
化合物177a： 1H NMR(400MHz,CD 3OD)δ7.41(d,J＝8.6Hz,2H),7.15–7.08(m,1H),6.91(d,J＝8.7Hz,2H),6.42–6.34(m,1H),4.63–4.58(m,1H),4.27(s,1H),3.97–3.89(m,1H),3.49–3.43(m,1H),3.37–3.32(m,2H),3.18–2.99(m,3H),2.73–2.61(m,4H),2.58–2.51(m,2H),2.43(s,3H),2.17–2.07(m,2H),1.91–1.82(m,2H),1.71–1.55(m,4H),1.40–1.34(m,2H),1.30(d,J＝6.0Hz,6H).LC-MS:ESI m/z 495.32[M+H] +,C 29H 42N 4O 3.计算值494.33.采用分析型手性SFC(ChiralPak IG(250×30mm)柱子)得到保留时间：0.760min，ee值：85.2％.
化合物177b： 1H NMR(400MHz,CD 3OD)δ7.54(d,J＝7.4Hz,1H),7.37(d,J＝8.3Hz,2H),6.91(d,J＝8.4Hz,2H),6.59(d,J＝7.3Hz,1H),4.63–4.57(m,1H),4.54–4.45(m,1H),3.84–3.74(m,1H),3.53–3.32(m,4H),3.17–2.93(m,4H),2.85–2.73(m,2H),2.70–2.67(m,2H),2.48(s,3H),2.21–2.07(m,2H),1.98–1.88(m,2H),1.80–1.62(m,4H),1.51–1.40(m,2H),1.30(d,J＝6.0Hz,6H).LC-MS:ESI m/z 495.3[M+H] +,C 29H 42N 4O 3.计算值494.33.采用分析型手性SFC(ChiralPak IG(250×30mm)柱子)得到保留时间：1.720min，ee值：87.6％.
化合物178(甲酸盐)： 1H NMR(400MHz,CD 3OD)δ7.57(d,J＝7.2Hz,1H),7.28(t,J＝7.9Hz,1H),7.08(s,1H),6.96(m,2H),6.61(d,J＝7.3Hz,1H),4.48(s,1H),4.00–3.74(m,5H),3.74–3.54(m,1H),3.53–3.31(m,5H),3.23–3.09(m,6H),2.81(t,J＝5.9Hz,2H),2.75–2.67(m,2H),2.47(d,J＝22.4Hz,3H),2.30–2.11(m,2H),1.98–1.90(m,2H),1.81–1.69(m,4H),1.50–1.40(m,2H).LC-MS:ESI m/z 522.3[M+H] +；C 30H 43N 5O 3计算值521.34.
化合物182(甲酸盐)： 1H NMR(400MHz,CD 3OD)δ7.48–7.39(m,3H),7.34–7.30(m,1H),7.20–7.18(m,1H),6.43(t,J＝8.0Hz,1H),4.30(s,0.5H),4.24(s,0.5H),3.87–3.76(m,1H), 3.40–3.31(m,4H),3.24–3.21(m,2H),3.09–3.05(m,2H),2.75–2.72(m,2H),2.60–2.53(m,2H),2.35–2.22(m,3H),2.15–2.09(m,2H),1.93–1.88(m,2H),1.72–1.63(m,4H),1.46–1.39(m,2H).LC-MS:ESI m/z 521.40[M+H] +,C 27H 35F 3N 4O 3.计算值520.27.
化合物184(甲酸盐)： 1H NMR(400MHz,CD 3OD)δ7.49–7.42(m,1H),7.25(t,J＝7.9Hz,1H),7.12–6.99(m,2H),6.91–6.82(m,1H),6.50–6.48(m,1H),4.33–4.28(m,1H),4.02(q,J＝7.0Hz,2H),3.95–3.77(m,1H),3.49–3.40(m,2H),3.30–2.98(m,4H),2.98–2.88(m,2H),2.80–2.72(m,2H),2.61(t,J＝7.6Hz,2H),2.48–2.30(m,3H),2.22–2.07(m,2H),1.96–1.86(m,2H),1.77–1.57(m,4H),1.50–1.38(m,2H),1.38–1.33(m,3H).LC-MS:ESI m/z 481.3[M+H] +；C 28H 40N 4O 3计算值480.31.
化合物186(甲酸盐)： 1H NMR(400MHz,CD 3OD)δ7.80–7.72(m,1H),7.72–7.62(m,1H),7.55(dd,J＝7.7,1.1Hz,1H),7.42(td,J＝7.7,2.4Hz,1H),7.32–7.27(m,1H),6.39(t,J＝7.1Hz,1H),4.29(s,0.5H),4.19(s,0.5H),3.73–3.68(m,1H),3.38–3.21(m,5H),3.15–3.02(m,3H),2.68–2.64(m,2H),2.56–2.50(m,2H),2.30–1.95(m,5H),1.90–1.76(m,2H),1.72–1.59(m,4H),1.44–1.34(m,2H).LC-MS:ESI m/z 462.4[M+H] +；C 27H 35N 5O 2计算值461.28.
化合物188(甲酸盐)： 1H NMR(400MHz,CD 3OD)δ7.50(d,J＝7.3Hz,1H),7.27(t,J＝7.9Hz,1H),7.17(s,1H),7.05(t,J＝9.0Hz,2H),6.53(d,J＝7.3Hz,1H),4.41–4.31(m,1H),3.94–3.79(m,1H),3.78–3.73(m,1H),3.47–3.43(m,2H),3.38–3.32(m,1H),3.30–3.26(m,1H),3.25–3.18(m,1H),3.16–3.04(m,1H),2.99(t,J＝7.6Hz,2H),2.78(t,J＝6.1Hz,2H),2.67–2.60(m,2H),2.44–2.36(m,3H),2.22–2.08(m,2H),1.95–1.89(m,2H),1.73–1.64(m,4H),1.46–1.38(m,2H),0.83–0.73(m,2H),0.70–0.60(m,2H).LC-MS:ESI m/z 493.2[M+H] +；C 29H 40N 4O 3计算值492.31.
化合物189(甲酸盐)： 1H NMR(400MHz,CD 3OD)δ8.52(s,1H),8.36–8.34(m,1H),7.86(d,J＝6.2Hz,1H),7.4–7.29(m,2H),6.41(dd,J＝7.2,5.5Hz,1H),4.34(s,0.5H),4.24(s,0.5H),3.75–3.64(m,1H),3.52–3.22(m,5H),3.16–2.94(m,3H),2.75–2.62(m,2H),2.58–2.52(m,2H),2.33–1.94(m,5H),1.89–1.76(m,2H),1.75–1.50(m,4H),1.45–1.32(m,2H).LC-MS:ESI m/z 438.3[M+H] +；C 25H 35N 5O 2.计算值437.28.
化合物190(甲酸盐)： 1H NMR(400MHz,CD 3OD)δ8.76(s,1H),8.11(d,J＝8.2Hz,1H),7.78(dd,J＝8.1,2.4Hz,1H),7.30(m,1H),6.44(dd,J＝9.9,7.3Hz,1H),4.47(s,0.5H),4.38(s,0.5H),3.83-3.76(m,1H),3.64–3.33(m,5H),3.29–3.02(m,3H),2.76–2.71(m,2H),2.63–2.56(m,2H),2.41–2.02(m,5H),1.98–1.84(m,2H),1.83–1.59(m,4H),1.53–1.37(m,2H). 19F NMR(376MHz,CD 3OD)δ-69.14(d,J＝2.6Hz,1H).LC-MS:ESI m/z 506.3[M+H] +；C 26H 34F 3N 5O 2计算值505.27.
化合物193(甲酸盐)： 1H NMR(400MHz,CD 3OD)δ9.05(s,1H),8.85(d,J＝2.6Hz,2H),7.46–7.43(m,1H),6.53–6.50(m,1H),4.45–4.38(m,1H),3.87–3.76(m,1H),3.58–3.50(m,1H),3.47–3.35(m,4H),3.25–3.11(m,2H),2.77(t,J＝6.0Hz,2H),2.65–2.61(m,2H),2.35–2.26(m,3H),2.22–2.14(m,2H),1.96–1.86(m,2H),1.83–1.65(m,4H),1.55–1.40(m,2H),1.38–1.25(m,1H).LC-MS:ESI m/z 439.5[M+H] +；C 24H 34N 6O 2计算值438.27.
化合物194(甲酸盐)： 1H NMR(400MHz,CD 3OD)δ7.62–7.58(m,1H),7.39–7.33(m,3H),7.18–7.13(m,1H),6.41–6.38(m,1H),4.19(s,0.5H),4.12(s,0.5H),3.78–3.68(m,1H),3.36–3.31(m,2H),3.27–3.22(m,2H),3.17–3.04(m,2H),3.02–2.96(m,2H),2.69–2.66(m,2H),2.54–2.49(m,2H),2.25(s,1.5H),2.18(s,1.5H),2.09–2.02(m,2H),1.83–1.80(m,2H),1.62–1.58(m,4H),1.38–1.31(m,2H).LC-MS:ESI m/z 515.3,5.17.3[M+H,Br] +,C 26H 35BrN 4O 2.计 算值:514.19.
化合物195(甲酸盐)： 1H NMR(400MHz,CD 3OD)δ7.51(d,J＝8.1Hz,2H),7.42(dd,J＝8.1,3.6Hz,2H),7.35–7.28(m,1H),6.49–6.41(m,1H),4.31–4.24(m,1H),3.79–3.75(m,1H),3.42–3.40(m,2H),3.18–2.93(m,4H),2.76–2.74(m,2H),2.59–2.54(m,2H),2.38–2.28(m,3H),2.22–2.08(m,2H),1.96–1.85(m,2H),1.78–1.63(m,4H),1.53–1.39(m,2H),1.37–1.25(m,2H).LC-MS:ESI m/z 515.16,517.16[M+H,Br] +；C 26H 35BrN 4O 2计算值514.19.
通过制备型手性SFC分离化合物195(15mg，0.03mmol)[柱：Daicel ChiralPak IG(250×30mm，10um)；流动相：含0.1％NH 3.H 2O的甲醇；梯度：60％甲醇，6.5min；流速：70g/min]，分别得到白色固体状的化合物195a和化合物195b。
化合物195a： 1H NMR(400MHz,CD 3OD)δ7.48(d,J＝8.5Hz,2H),7.41(d,J＝8.5Hz,2H),7.12(d,J＝7.3Hz,1H),6.35(d,J＝7.3Hz,1H),4.17(s,1H),3.84–3.78(m,1H),3.39–3.33(m,2H),3.23–3.03(m,4H),2.95–2.86(m,2H),2.68(t,J＝6.2Hz,1H),2.52(t,J＝7.5Hz,1H),2.26(s,3H),2.11(dd,J＝14.7,7.2Hz,2H),1.89–1.82(m,2H),1.71–1.60(m,4H),1.43–1.30(m,4H).LC-MS:ESI m/z 515.2/517.2[M+H,Br] +C 26H 35BrN 4O 2计算值514.19.采用分析型手性SFC(ChiralPak IG(250×30mm)柱子)得到保留时间：2.669min，ee值：96.9％.
化合物195b： 1H NMR(400MHz,CD 3OD)δ7.49(d,J＝8.4Hz,2H),7.41(d,J＝8.5Hz,2H),7.16(d,J＝7.4Hz,1H),6.37(d,J＝7.3Hz,1H),4.22(s,1H),3.68(s,1H),3.40–3.34(m,2H),3.13–2.94(m,4H),2.69(t,J＝6.2Hz,2H),2.57–2.51(m,2H),2.37(s,3H),2.12–2.04(m,2H),1.91–1.83(m,2H),1.71–1.60(m,4H),1.44–1.30(m,4H).LC-MS:ESI m/z 515.2/517.2[M+H,Br] +C 26H 35BrN 4O 2计算值514.19.采用分析型手性SFC(ChiralPak IG(250×30mm)柱子)得到保留时间：4.507min，ee值：96.1％.
化合物196(甲酸盐)： 1H NMR(400MHz,CD 3OD)δ7.65–7.57(m,2H),7.46(dd,J＝7.3,0.9Hz,1H),7.33(t,J＝7.5Hz,1H),7.22–7.14(m,1H),6.50(d,J＝7.3Hz,1H),4.79–4.70(m,1H),3.99–3.85(m,1H),3.49–3.41(m,3H),3.39–3.32(m,2H),3.30–3.21(m,1H),3.16–3.06(m,2H),2.77(t,J＝6.1Hz,2H),2.63(t,J＝7.6Hz,2H),2.41–2.31(m,3H),2.27–2.19(m,1H),2.17–2.05(m,1H),1.95–1.86(m,2H),1.79–1.63(m,4H),1.49–1.37(m,2H).LC-MS:ESI m/z 515.2,517.0[M+H,Br] +；C 26H 35BrN 4O 2计算值514.19.
化合物200(甲酸盐)： 1H NMR(400MHz,CD 3OD)δ7.70–7.61(m,1H),7.40(t,J＝7.7Hz,1H),7.26–7.19(m,1H),7.13–7.03(m,1H),6.52–6.45(m,1H),4.76–4.67(m,1H),3.94–3.81(m,1H),3.50–3.40(m,3H),3.39–3.32(m,2H),3.27–3.19(m,1H),3.17–3.07(m,2H),2.80–2.72(m,2H),2.67–2.57(m,2H),2.41–2.28(m,3H),2.26–2.03(m,2H),1.95–1.86(m,2H),1.82–1.65(m,4H),1.54–1.38(m,2H).LC-MS:ESI m/z 489.2,491.2[M+H,Cl] +；C 26H 34ClFN 4O 2计算值488.24.
化合物201(甲酸盐)： 1H NMR(400MHz,CD 3OD)δ7.61–7.50(m,1H),7.46(m,1H),7.37–7.31(m,1H),7.18(t,J＝7.5Hz,1H),7.15–7.07(m,1H),6.51(d,J＝7.3Hz,1H),4.73(s,1H),3.79–3.74(m,1H),3.47–3.34(m,4H),3.30–3.02(m,4H),2.77(t,J＝6.2Hz,2H),2.65(t,J＝7.8Hz,2H),2.41(s,1.5H),2.32(s,1.5H),2.20–2.11(m,2H),1.95–1.85(m,2H),1.81–1.60(m,4H),1.48–1.43(m,2H). 19F NMR(376MHz,CD 3OD)δ-116.94,-117.70.LC-MS:ESI m/z 455.3[M+H] +；C 26H 35FN 4O 2计算值454.27.
化合物202(甲酸盐)： 1H NMR(400MHz,CD 3OD)δ7.72(m,2H),7.65–7.61(m,1H),7.48–7.37(m,2H),6.50(m,1H),4.71(s,0.5H),4.65(s,0.5H),3.92–3.84(m,1H),3.61–3.35(m,6H), 3.18(m,2H),2.79–2.73(m,2H),2.66–2.59(m,2H),2.40–2.26(m,5H),1.93–1.89(m,2H),1.83–1.63(m,4H),1.56–1.43(m,2H).LC-MS:ESI m/z 462.3[M+H] +；C 27H 35N 5O 2计算值461.28.
化合物203(甲酸盐)： 1H NMR(400MHz,CD 3OD)δ7.48(d,J＝7.3Hz,1H),7.45–7.35(m,3H),7.06–6.98(m,1H),6.52(d,J＝7.3Hz,1H),5.01–4.93(m,1H),3.95–3.84(m,2H),3.45–3.40(m,2H),3.38–3.31(m,1H),3.29–3.21(m,1H),2.95–2.72(m,6H),2.69–2.63(m,2H),2.49–2.46(m,3H),2.25–2.12(m,2H),1.94–1.88(m,2H),1.75–1.59(m,4H),1.51–1.39(m,2H),0.88–0.73(m,4H).LC-MS:ESI m/z 493.3[M+H] +；C 29H 40N 4O 3计算值492.31.
化合物204(甲酸盐)： 1H NMR(400MHz,CD 3OD)δ7.48(d,J＝7.3Hz,1H),7.46–7.36(m,2H),7.12–7.06(m,1H),7.06–6.99(m,1H),6.52(d,J＝7.3Hz,1H),5.14–5.13(m,1H),4.29–4.11(m,2H),4.09–3.94(m,1H),3.86–3.70(m,2H),3.46–3.42(m,2H),3.41–3.34(m,2H),3.34–3.20(m,3H),2.98–2.89(m,1H),2.85–2.63(m,7H),2.54–2.53(m,3H),2.32–2.15(m,2H),1.95–1.87(m,2H),1.76–1.59(m,4H),1.51–1.39(m,2H).LC-MS:ESI m/z 511.3[M+H] +；C 29H 42N 4O 4计算值510.32.
化合物205(甲酸盐)： 1H NMR(400MHz,CD 3OD)δ7.50(d,J＝7.3Hz,1H),7.45(d,J＝7.6Hz,1H),7.26(d,J＝4.1Hz,2H),7.19–7.15(m,1H),6.54(d,J＝7.3Hz,1H),4.68(s,1H),3.87–3.80(m,1H),3.46–3.38(m,4H),3.30–3.27(m,1H),3.06–3.01(m,3H),2.84–2.74(m,4H),2.68–2.65(m,2H),2.46(s,3H),2.11–1.89(m,4H),1.75–1.67(m,4H),1.48–1.41(m,2H),1.46(t,J＝7.5Hz,3H).LC-MS:ESI m/z 465.2[M+H] +；C 28H 40N 4O 2计算值464.32.
化合物207(甲酸盐)： 1H NMR(400MHz,CD 3OD)δ7.67–7.64(m,1H),7.45–7.29(m,4H),6.55-6.49(m,1H),4.74(s,0.6H),4.68(s,0.4H),3.90–3.35(m,6H),3.25–3.07(m,3H),2.78–2.74(m,2H),2.69–2.59(m,2H),2.45(s,1.2H),2.28(s,1.8H),2.22–2.03(m,2H),1.93–1.87(m,2H),1.76–1.65(m,4H),1.60–1.46(m,2H). 19F NMR(376MHz,CD 3OD)δ-57.57.LC-MS:ESI m/z 521.3[M+H] +；C 27H 35F 3N 4O 3计算值520.27.
化合物208(甲酸盐)： 1H NMR(400MHz,CD 3OD)δ7.47–7.36(m,3H),7.10–7.05(m,1H),7.03-6.99(m,1H),6.52–6.47(m,1H),5.23(s,0.5H),5.10(s,0.5H),4.04–3.96(m,1H),3.92–3.86(m,1H),3.83–3.78(m,1H),3.49–3.38(m,3H),3.21–3.12(m,1H),2.78–2.72(m,3H),2.71–2.60(m,4H),2.52(s,1.5H),2.50(s,1.5H),2.31–2.07(m,4H),1.92–1.86(m,2H),1.72–1.46(m,6H),1.08(t,J＝6.2Hz,6H).LC-MS:ESI m/z 509.4[M+H] +；C 30H 44N 4O 3计算值508.34.
化合物211(甲酸盐)： 1H NMR(400MHz,CD 3OD)δ7.47–7.36(m,3H),7.11–6.92(m,2H),6.52(d,J＝7.3Hz,1H),5.33(s,0.5H),5.21(s,0.5H),4.11–3.85(m,3H),3.49–3.32(m,3H),3.26–3.10(m,1H),2.80–2.73(m,3H),2.76(t,J＝6.0Hz,2H),2.65(t,J＝7.8Hz,2H),2.53(s,1.5H),2.52(s,1.5H),2.34–2.18(m,2H),1.93–1.87(m,2H),1.84–1.44(m,6H),1.43–1.24(m,2H),0.65–0.61(m,2H),0.42–0.38(m,2H).LC-MS:ESI m/z 507.3[M+H] +；C 30H 42N 4O 3计算值506.33.
化合物212(甲酸盐)： 1H NMR(400MHz,CD 3OD)δ7.98–7.83(m,2H),7.71–7.63(m,1H),7.60–7.52(m,1H),7.44–7.36(m,1H),6.54–6.48(m,1H),5.42–5.36(m,1H),4.06–3.97(m,1H),3.75–3.63(m,1H),3.46–3.40(m,3H),3.16–2.94(m,4H),2.88(s,3H),2.87(s,3H),2.79–2.73(m,2H),2.70–2.63(m,2H),2.49(s,1H),2.37(s,2H),2.25–2.18(m,1H),2.10–2.01(m,1H),1.95–1.87(m,2H),1.77–1.69(m,4H),1.59–1.40(m,2H).LC-MS:ESI m/z 544.2[M+H] +；C 28H 41N 5O 4S计算值543.29.
化合物213(甲酸盐)： 1H NMR(400MHz,CD 3OD)δ7.48(d,J＝7.3Hz,1H),7.44–7.32(m,2H),7.00(t,J＝7.6Hz,1H),6.94–6.89(m,1H),6.53(d,J＝7.3Hz,1H),5.22(s,0.5H),5.10(s,0.5H),4.80–4.73(m,1H),3.97–3.91(m,1H),3.74–3.57(m,1H),3.46–3.41(m,2H),3.37–3.31(m,1H),3.27–3.13(m,1H),2.86–2.56(m,8H),2.51–2.50(m,4H),2.25–2.17(m,4H),1.93–1.81(m,3H),1.74–1.46(m,7H).LC-MS:ESI m/z 507.3[M+H] +；C 30H 42N 4O 3计算值506.33.
化合物214(甲酸盐)： 1H NMR(400MHz,CD 3OD)δ7.54–7.32(m,3H),7.09-7.05(m,1H),7.01–6.96(m,1H),6.52(d,J＝7.3Hz,1H),5.20(s,0.4H),5.06(s,0.6H),4.96–4.92(m,1H),4.02–3.93(m,1H),3.58–3.32(m,3H),3.29–3.08(m,2H),2.82–2.61(m,7H),2.51(s,1.2H),2.50(s,1.8H),2.24–2.18(m,2H),2.05–1.75(m,8H),1.71–1.28(m,8H).LC-MS:ESI m/z 521.3[M+H] +；C 31H 44N 4O 3计算值520.34.
化合物215(甲酸盐)： 1H NMR(400MHz,CD 3OD)δ7.47–7.36(m,3H),7.08–7.01(m,2H),6.53–6.51(m,1H),5.15–5.04(m,2H),4.06–3.82(m,5H),3.49–3.42(m,3H),3.25–3.12(m,1H),2.83–2.65(m,10H),2.36–2.13(m,4H),1.94–1.88(m,2H),1.75–1.59(m,4H),1.54–1.50(m,2H),1.33–1.29(m,1H).LC-MS:ESI m/z 523.3[M+H] +；C 30H 42N 4O 4计算值522.32.
化合物216(甲酸盐)： 1H NMR(400MHz,CD 3OD)δ8.00–7.96(m,1H),7.86–7.80(m,1H),7.59–7.57(m,1H),7.48–7.43(m,1H),7.35–7.32(m,1H),6.43–6.40(m,1H),5.40(s,0.5H),5.35(s,0.5H),3.84–3.80(m,1H),3.48–3.38(m,1H),3.34–3.32(m,4H),3.28(s,3H),3.15–2.91(m,3H),2.66(t,J＝6.1Hz,2H),2.56–2.53(m,2H),2.31(s,1.5H),2.22(s,1.5H),2.14–1.94(m,2H),1.83–1.78(m,2H),1.66–1.59(m,4H),1.39–1.32(m,2H).LC-MS:ESI m/z 515.3[M+H] +；C 27H 38N 4O 4S计算值514.26.
化合物217(甲酸盐)： 1H NMR(400MHz,CD 3OD)δ7.60–7.55(m,1H),7.45–7.39(m,1H),7.36–7.30(m,2H),7.29–7.23(m,1H),7.223–7.07(m,2H),7.06–7.03(m,1H),7.01–6.98(m,1H),6.92–6.86(m,1H),6.51–6.44(m,1H),4.96(s,0.5H),4.89(s,0.5H),4.04–3.85(m,1H),3.63–3.31(m,3H),3.12–3.04(m,1H),2.98–2.83(m,3H),2.78–2.71(m,2H),2.68–2.58(m,2H),2.54(s,1.5H),2.46(s,1.5H),2.43–2.24(m,1H),2.20–2.02(m,2H),1.93–1.84(m,2H),1.79–1.54(m,4H),1.52–1.38(m,2H).LC-MS:ESI m/z 529.3[M+H] +；C 32H 40N 4O 3计算值528.31.
化合物221(甲酸盐)： 1H NMR(400MHz,CD 3OD)δ7.47–7.36(m,3H),7.08–7.01(m,2H),6.53–6.51(m,1H),5.15–5.04(m,2H),4.06–3.82(m,5H),3.49–3.42(m,3H),3.25–3.12(m,1H),2.83–2.65(m,10H),2.36–2.13(m,4H),1.94–1.88(m,2H),1.75–1.59(m,4H),1.54–1.50(m,2H),1.33–1.29(m,1H).LC-MS:ESI m/z 523.3[M+H] +；C 30H 42N 4O 4计算值522.32.
实施例34 化合物224和化合物225的合成
Figure PCTCN2021078537-appb-000191
步骤1：6-甲氧基烟酸
Figure PCTCN2021078537-appb-000192
向6-氯烟酸(2.0g，12.7mmol)在DMSO(25mL)的溶液中加入甲醇(1.03mL，25.4mmol)。将反应混合物在100℃加热18小时并冷却至室温。加入1N盐酸溶液调节pH＝3-4。过滤收集得到的沉淀，用水洗涤并真空干燥，得到标题化合物(1.06g，54％)，为白色固体。 1H NMR(400MHz,DMSO-d 6)δ13.04(s,1H),8.73(d,J＝2.2Hz,1H),8.14(dd,J＝8.7,2.4Hz,1H),6.91(d,J＝8.7Hz,1H),3.93(s,3H).
步骤2：2-重氮-1-(6-甲氧基吡啶-3-基)乙-1-酮
Figure PCTCN2021078537-appb-000193
在0℃下，向6-乙氧基烟酸(1.0g，6.53mmol)的二氯甲烷(20mL)溶液中加入草酰氯(5.6mL，65.3mmol)和两滴DMF。将混合物在室温搅拌2小时。真空除去溶剂。将残余物溶于二氯甲烷(20mL)。然后缓慢加入三甲基硅烷化重氮甲烷的己烷溶液(2N，6.55mL，13.1mmol)和三乙胺(1.82mL，13.1mmol)，并将所得溶液在0℃下搅拌3小时，后升到室温再搅拌12小时。然后将反应过滤并减压浓缩，得到标题化合物(780mg，67％)，为棕色固体。LC-MS:ESI m/z:178.2.[M+H] +；C 8H 7N 3O 2 require 177.05.
步骤3：2-(6-甲氧基吡啶-3-基)乙酸甲酯
Figure PCTCN2021078537-appb-000194
向2-重氮-1-(6-甲氧基吡啶-3-基)乙-1-酮(780mg，4.4mmol)的甲醇(20mL)溶液中加入Ag 2O(1.02g，4.4mmol)。将混合物在65℃加热16小时。然后将反应冷却至室温，过滤并减压浓缩。将残余物通过快速柱色谱纯化(0-30％乙酸乙酯的石油醚溶液)，得到标题化合物(300mg，37％)，为浅黄色油状物。LC-MS:ESI m/z:182.1.[M+H] +；C 9H 11NO 3 require 181.07.
步骤4：2-溴-2-(6-甲氧基吡啶-3-基)乙酸甲酯
Figure PCTCN2021078537-appb-000195
在N 2气氛下，向2-(6-甲氧基吡啶-3-基)乙酸甲酯(100mg，0.55mmol)在CCl 4(4mL)的溶液中加入NBS(103mg，0.58mmol)和AIBN(13mg，0.08mmol)。将混合物在80℃下加热16小时。将混合物过滤并将滤液真空浓缩。将残余物通过快速柱色谱法纯化(0-30％乙酸乙酯的石油醚溶液)，得到标题化合物(120mg，84％)，为黄色油状物。LC-MS:ESI m/z:260.1,262.1.[M+H,Br] +；C 9H 10BrNO 3 require 258.98.
步骤5：7-(5-((3S)-3-((2-甲氧基-1-(6-甲氧基吡啶-3-基)-2-氧代乙基)(甲基)氨基)吡咯烷-1-基)戊基)-3,4-二氢-1,8-萘啶-1(2H)-羧酸叔丁酯
Figure PCTCN2021078537-appb-000196
向2-溴-2-(6-甲氧基吡啶-3-基)乙酸甲酯(80mg，0.31mmol)的乙腈(4mL)溶液中加入(S)-7-(5-(3-(甲基氨基)吡咯烷-1-基)戊基)-3,4-二氢-1,8-萘啶-1(2H)-羧酸叔丁酯(125mg，0.31mmol)和K 2CO 3(86mg，0.62mmol)。将混合物在室温搅拌16小时。真空除去溶剂。通过快速柱色谱法纯化(0-4％甲醇的二氯甲烷溶液)，得到标题化合物(100mg，55％)，为黄色油状物。LC-MS:ESI m/z:582.6.[M+H] +；C 32H 47N 5O 5 require 581.36.
步骤6：化合物224和化合物225
在室温下将7-(5-((3S)-3-((2-甲氧基-1-(6-甲氧基吡啶-3-基)-2-氧代乙基)(甲基)氨基)吡咯烷-1-基)戊基)-3,4-二氢-1,8-萘啶-1(2H)-羧酸叔丁酯(100mg，0.18mmol)的盐酸/1，4-二氧六环(4N,4mL)溶液搅拌2小时。真空浓缩溶剂。将残余物溶于甲醇(4mL)和H 2O(1mL)中，然后加入LiOH(21mg，0.88mmol)。将混合物在40℃下搅拌2小时。通过甲酸将混合物调节至pH≈7，真空浓缩溶剂。将残余物通过制备型HPLC在以下条件下纯化[柱：Kromasil Prep C18，30×150mm；流动相：含0.1％甲酸的1-40％乙腈水溶液]，得到白色固体的化合物224(17.15mg，21％)。 1H NMR(400MHz,CD 3OD)δ8.17(s,1H),7.82–7.78(m,1H),7.5–7.42(m,1H),6.79(d,J＝8.6Hz,1H),6.50(dd,J＝7.3,3.4Hz,1H),4.38(s,0.5H),4.30(s,0.5H),3.89(s,3H),3.80–3.40(m,5H),3.28–3.01(m,4H),2.83–2.72(m,2H),2.66–2.62(m,2H),2.39(s,1.5H),2.28(s,1.5H),2.22–2.06(m,2H),1.94–1.90(m,2H),1.80–1.63(m,4H),1.51–1.42(m,2H).LC-MS:ESI m/z 468.4[M+H] +；C 26H 37N 5O 3计算值467.29. 以及得到化合物225(15.4mg，19％)为白色固体。 1H NMR(400MHz,CD 3OD)δ7.75–7.69(m,1H),7.48–7.44(m,2H),6.62–6.45(m,2H),4.17(s,0.5H),4.10(s,0.5H),3.83–3.75(m,1H),3.60–3.32(m,5H),3.26–3.04(m,3H),2.77(t,J＝5.8Hz,2H),2.71–2.61(m,2H),2.39–2.05(m,5H),1.96–1.86(m,2H),1.82–1.61(m,4H),1.50–1.43(m,2H).LC-MS:ESI m/z 454.3[M+H] +；C 25H 35N 5O 3计算值453.27.
实施例35.参照化合物224的合成方法制备得到以下化合物：
化合物226(甲酸盐)： 1H NMR(400MHz,CD 3OD)δ8.15(s,1H),7.85–7.72(m,1H),7.46(dd,J＝7.3,3.2Hz,1H),6.77(d,J＝8.6Hz,1H),6.51(dd,J＝7.3,2.8Hz,1H),4.44–4.20(m,3H),3.83–3.41(m,5H),3.28–2.93(m,4H),2.77(t,J＝6.0Hz,2H),2.65(t,J＝7.8Hz,2H),2.39(s,1.6H),2.29(s,1.4H),2.22–2.12(m,2H),1.98–1.85(m,2H),1.83–1.60(m,4H),1.50-1.43(m,2H),1.36(t,J＝8.0Hz,3H).LC-MS:ESI m/z 482.3[M+H] +；C 27H 39N 5O 3 require 481.31.
化合物227(甲酸盐)： 1H NMR(400MHz,CD 3OD)δ8.15(s,1H),7.79–7.74(m,1H),7.44–7.37(m,1H),6.75–6.70(m,1H),6.52–6.47(m,1H),5.23–5.17(m,1H),4.43–4.39(s,0.5H),4.35–4.30(s,0.5H),3.80–3.73(m,1H),3.42(m,3H),3.22–2.94(m,5H),2.79–2.72(m,2H),2.67–2.59(m,2H),2.42(s,1.5H),2.31(s,1.5H),2.21–2.10(m,2H),1.94–1.86(m,2H),1.78–1.63(m,4H),1.54–1.43(m,2H),1.31(d,J＝6.1Hz,6H).LC-MS:ESI m/z 496.4[M+H] +；C 28H 41N 5O 3计算值495.32.
化合物228(甲酸盐)： 1H NMR(400MHz,CD 3OD)δ7.63(t,J＝7.3Hz,1H),7.47(t,J＝7.0Hz,1H),7.27(d,J＝9.5Hz,2H),6.55–6.51(m,1H),4.96(s,0.5H),4.89(s,0.5H),4.83–4.77(m,1H),3.92–3.84(m,1H),3.58–3.39(m,4H),3.15–2.99(m,4H),2.78(t,J＝6.0Hz,2H),2.71–2.66(m,2H),2.51(s,1.5H),2.42(s,1.5H),2.32–2.12(m,2H),1.95–1.89(m,2H),1.83–1.68(m,4H),1.57–1.48(m,2H),1.44–1.36(m,6H).LC-MS:ESI m/z 563.2[M+H] +；C 30H 41F 3N 4O 3计算值562.31.
化合物229(甲酸盐)： 1H NMR(400MHz,CD 3OD)δ7.52–7.37(m,3H),7.21–7.13(m,1H),7.08–7.04(m,1H),6.52–6.49(m,1H),5.20–5.11(m,1H),4.73–4.71(m,1H),4.02–3.55(m,6H),3.44–3.41(m,2H),3.24–3.12(m,1H),2.91–2.48(m,10H),2.23–2.03(m,3H),1.93–1.89(m,2H),1.70–1.62(m,4H),1.50–1.44(m,2H).LC-MS:ESI m/z 545.3,547.2[M+H,Cl] +；C 29H 41ClN 4O 4计算值544.28.
实施例36.化合物230的合成
Figure PCTCN2021078537-appb-000197
步骤1：3,5-二甲基-1-(3-硝基苯基)-1H-吡唑
Figure PCTCN2021078537-appb-000198
在N 2气氛下，将3-硝基苯肼盐酸盐(25g，132mmol)和戊烷-2,4-二酮(8.8g，88mmol)加入到AcOH(90mL)溶液中，然后加入三乙胺(13.3g，132mmol)。将混合物在25℃下搅拌16小时。将混合物减压浓缩，并将残余物溶解在乙酸乙酯(300mL)中，并用水(400mL)，盐水洗涤并经硫酸钠干燥。真空除去溶剂，并通过快速柱色谱法纯化(0-80％乙酸乙酯的石油醚溶液)，得到标题化合物(17.7g，93％)，为黄色固体。 1H NMR(400MHz,DMSO-d 6)δ8.31(t,J＝2.1Hz,1H),8.22–8.18(m,1H),8.03–8.00(m,1H),7.78(t,J＝8.2Hz,1H),6.16(s,1H),2.40(s,3H),2.21(s,3H).LC-MS:ESI m/z 218.1[M+H] +；C 11H 11N 3O 2计算值217.09.
步骤2：3-(3,5-二甲基-1H-吡唑-1-基)苯胺
Figure PCTCN2021078537-appb-000199
在1大气压的H 2气氛下，向3，5-二甲基-1-(3-硝基苯基)-1H-吡唑(2.17g，1mmol)的乙酸乙酯(20mL)溶液中加入10％的Pd/C(200mg)。在25℃下搅拌16小时后，将混合物通过硅藻土过滤，并将硅藻土用乙酸乙酯洗涤。减压蒸发合并的有机物，得到标题化合物(1.88g，100％)，为黄色油状物。 1H NMR(400MHz,DMSO-d 6)δ7.10–7.06(m,1H),6.66(t,J＝2.1Hz,1H),6.56–6.52(m,2H),6.00(s,1H),2.25(s,3H),2.15(s,3H).LC-MS:ESI m/z 188.2[M+H] +；C 11H 13N 3计算值187.11.
步骤3：3-((3-(3,5-二甲基-1H-吡唑-1-基)苯基)氨基甲酰基)吡咯烷-1-甲酸叔丁酯
Figure PCTCN2021078537-appb-000200
在N 2气氛下，向3-(3,5-二甲基-1H-吡唑-1-基)苯胺(1.7g，9.1mmol)和1-(叔丁氧羰基)吡咯烷-3-羧酸(2.9g，13.6mmol)的二氯甲烷(30mL)溶液中加入三乙胺(2.7g，27.3mmol)和T3P(4.3g，13.6mmol)。在25℃下搅拌16小时后，将反应混合物用NaHCO 3(40mL)稀释，并用二氯甲烷(15mL×3)萃取。合并的有机相用盐水洗涤，经硫酸钠干燥，并过滤。将滤液在减压下浓缩。残余物通过快速柱色谱法纯化(0-70％乙酸乙酯的石油醚溶液)，得到标题化合物(3.38g，97％)，为白色固体。 1H NMR(400MHz,DMSO-d 6)δ10.23(s,1H),7.85(s,1H),7.53(d,J＝9.0Hz,1H),7.43–7.38(m,1H),7.17(d,J＝7.9Hz,1H),6.07(s,1H),3.55–3.50(m,1H),3.41–3.36(m,2H),3.29–3.23(m,1H),3.17–3.10(m,1H),2.30(s,3H),2.17(s,3H),2.14–2.08(m,1H),2.03–1.98(m,1H),1.40(s,9H).LC-MS:ESI m/z 385.3[M+H] +；C 21H 28N 4O 3计算值384.22
步骤4：3-((3-(3,5-二甲基-1H-吡唑-1-基)苯基)(2-甲氧基-2-氧乙基)氨基甲酰基)吡咯烷-1-甲酸叔丁酯
Figure PCTCN2021078537-appb-000201
在0℃，N 2气氛下，向NaH(484mg，在矿物油中的60％的分散液)在四氢呋喃(30mL)的悬浮液中加入3-((3-(3,5-二甲基-1H-吡唑-1-基)苯基)氨基甲酰基)吡咯烷-1-甲酸叔丁酯(3.1g，8.1mmol)。升到25℃搅拌15分钟后，将2-溴乙酸甲酯(1mL，8.9mmol)加入到反应混合物中。在25℃下进一步搅拌2小时后，将反应混合物用NH 4Cl(40mL)淬灭，并用二氯甲烷(15mL×3)萃取。合并的有机相用盐水洗涤，经硫酸钠干燥，并过滤。将滤液在减压下浓缩。残余物通过快速柱色谱法纯化(0-70％乙酸乙酯的石油醚溶液)，得到标题化合物(3.27g，86％)，为黄色油状物。 1H NMR(400MHz,DMSO-d 6)δ7.61–7.55(m,3H),7.44–7.39(m,1H),6.10(s,1H),4.39(s,2H),3.66(s,3H),3.31–3.27(m,1H),3.25(d,J＝7.3Hz,2H),3.12–3.03(m,2H),2.34(s,3H),2.18(s,3H),1.99–1.91(m,1H),1.82(s,1H),1.36(s,9H).LC-MS:ESI m/z 457.3[M+H] +；C 24H 32N 4O 5计算值456.24.
步骤5-8参照化合物97(步骤8-11)的合成方法，制备得到化合物230：
1H NMR(400MHz,DMSO-d 6)δ7.61–7.50(m,2H),7.39(d,J＝7.3Hz,1H),7.01(d,J＝7.2Hz,1H),6.48(s,1H),6.24(d,J＝7.1Hz,1H),6.09(s,1H),4.18(s,2H),3.22(s,2H),2.98–2.90(m,1H),2.71–2.51(m,8H),2.48–2.37(m,2H),2.31(s,3H),2.17(s,3H),1.99–1.91(m,1H),1.78–1.58(m,3H).LC-MS:ESI m/z 503.3[M+H] +；C 28H 34N 6O 3计算值502.27.
实施例37.在以下条件下通过制备型手性SFC拆分化合物230：[柱：Daicel ChiralPak IG(250*30mm，10um)；流动相：0.1％NH 3.H 2O的甲醇溶液；梯度：60％甲醇，6.5min；流动速度：70g/min]分别得到化合物230a和化合物230b，同时参照化合物230的合成方法，制备得到化合物231，化合物231a，化合物231b，化合物232，化合物232a，化合物232b
化合物230a： 1H NMR(400MHz,DMSO-d 6)δ7.63–7.45(m,2H),7.39(d,J＝6.9Hz,1H),7.02(d,J＝7.3Hz,1H),6.49(s,1H),6.24(d,J＝7.3Hz,1H),6.09(s,1H),4.21(s,2H),3.24–3.20(m,2H),3.01–2.87(m,2H),2.72–2.52(m,8H),2.46–2.40(m,1H),2.32(s,3H),2.18(s,3H),1.99–1.92(m,1H),1.77–1.69(m,2H),1.68–1.60(m,1H).LC-MS:ESI m/z 503.3[M+H] +；C 28H 34N 6O 3计算值502.27.
化合物230b： 1H NMR(400MHz,DMSO-d 6)δ7.63–7.57(m,3H),7.55–7.32(m,2H),6.53(s,1H),6.11(s,1H),4.33(q,J＝17.4Hz,2H),3.65–3.36(m,8H),3.12–2.96(m,2H),2.89(td,J＝14.4,7.3Hz,1H),2.72–2.65(m,2H),2.35(s,3H),2.18(s,3H),2.14–1.92(m,2H),1.83–1.75(m,2H).LC-MS:ESI m/z 503.3[M+H] +；C 28H 34N 6O 3计算值502.27.
化合物231(甲酸盐)： 1H NMR(400MHz,DMSO-d 6)δ7.66–7.54(m,3H),7.46–7.40(m,1H),7.23(br,1H),6.39(br,1H),6.11(s,1H),4.38–4.27(m,2H),3.39–3.20(m,7H),3.13–3.07(m,2H),2.64(t,J＝5.9Hz,2H),2.61–2.52(m,2H),2.34(s,3H),2.18(s,3H),2.09–2.01(m,1H),1.97–1.84(m,3H),1.80–1.73(m,2H).LC-MS:ESI m/z 517.3[M+H] +；C 29H 36N 6O 3计算值516.28.
将外消旋混合物化合物231(41.14mg，0.080mmol)在以下条件下通过手性制备SFC分离[柱：Daicel ChiralPak IG(250*30mm，10um)；流动相：含0.1％NH 3·H 2O的甲醇溶液；梯度：60％甲醇，6.5min；流速：70g/min]，分别得到化合物231a和化合物231b，为白色固体。
化合物231a： 1H NMR(400MHz,CD 3OD)δ7.68–7.57(m,2H),7.56–7.48(m,2H),7.24(d,J＝7.3Hz,1H),6.42(d,J＝7.3Hz,1H),6.09(s,1H),4.33(d,J＝16.7Hz,1H),4.18(d,J＝16.7Hz,1H),3.84(dd,J＝11.6,3.3Hz,1H),3.53–3.44(m,2H),3.44–3.38(m,2H),3.21–3.00(m,4H),2.80–2.67(m,4H),2.40–2.33(m,1H),2.31(s,3H),2.25(s,3H),2.20–2.13(m,1H),2.03–1.97(m,2H),1.92–1.83(m,2H).LC-MS:ESI m/z 517.3[M+H] +,C 29H 36N 6O 3.计算值516.28.
化合物231b： 1H NMR(400MHz,CD 3OD)δ7.67–7.58(m,2H),7.52(t,J＝8.9Hz,2H),7.27(d,J＝7.3Hz,1H),6.43(d,J＝7.3Hz,1H),6.10(s,1H),4.36(d,J＝16.7Hz,1H),4.17(d,J＝16.8Hz,1H),3.85(dd,J＝11.5,3.5Hz,1H),3.53–3.45(m,2H),3.45–3.39(m,2H),3.22–3.03(m,4H),2.74(dd,J＝13.6,6.8Hz,4H),2.40–2.33(m,1H),2.31(s,3H),2.25(s,3H),2.19–2.12(m,1H),2.05–1.98(m,2H),1.92–1.84(m,2H).LC-MS:ESI m/z 517.3[M+H] +,C 29H 36N 6O 3.计算值516.28.
化合物232(甲酸盐)： 1H NMR(400MHz,CD 3OD)δ7.68–7.58(m,2H),7.54–7.48(m,2H),7.39(d,J＝7.3Hz,1H),6.48(d,J＝7.3Hz,1H),6.10(s,1H),4.50(d,J＝16.8Hz,1H),3.98(d,J＝16.8Hz,1H),3.82–3.78(m,1H),3.59–3.50(m,2H),3.45–3.40(m,2H),3.33–3.32(m,1H),3.26–3.22(m,1H),3.18–3.11(m,2H),2.76(t,J＝6.1Hz,2H),2.70–2.63(m,2H),2.48–2.41(m,1H),2.32(s,3H),2.24(s,3H),2.13–2.07(m,1H),1.93–1.87(m,2H),1.84–1.62(m,4H).LC-MS:ESI m/z 531.1[M+H] +；C 30H 38N 6O 3计算值530.30.
将化合物232外消旋混合物(17.92mg，0.034mmol)在以下条件下通过手性制备SFC分离[柱：Daicel ChiralPak IG(250×30mm，10um)；流动相：含0.1％NH 3·H 2O的甲醇溶液；梯度：60％甲醇，6.5min；流速：70g/min]，分别得到化合物232a和化合物232b，为白色固体。
化合物232a： 1H NMR(400MHz,CD 3OD)δ7.62(dd,J＝15.9,8.2Hz,2H),7.51(dd,J＝17.0,8.0Hz,2H),7.25(d,J＝7.1Hz,1H),6.42(d,J＝7.3Hz,1H),6.09(s,1H),4.40(d,J＝16.7Hz,1H),4.04(d,J＝17.1Hz,1H),3.72(d,J＝11.5Hz,1H),3.49–3.43(m,2H),3.42–3.37(m,2H),3.17–3.13(m,1H),3.07–3.01(m,2H),2.75–2.69(m,2H),2.64–2.57(m,2H),2.40–2.36(m,1H),2.31(s,3H),2.24(s,3H),2.06–2.00(m,2H),1.91–1.85(m,2H),1.78–1.62(m,4H).LC-MS:ESI m/z 531.3[M+H] +,C 30H 38N 6O 3.计算值530.30.
化合物232b： 1H NMR(400MHz,CD 3OD)δ7.63(dd,J＝15.8,7.9Hz,2H),7.52(dd,J＝16.7,8.1Hz,2H),7.32(d,J＝7.3Hz,1H),6.45(d,J＝7.3Hz,1H),6.10(s,1H),4.46(d,J＝16.4Hz,1H),4.00(d,J＝16.8Hz,1H),3.76(d,J＝11.3Hz,1H),3.54–3.46(m,2H),3.44–3.38(m,2H),3.23–3.17(m,1H),3.13–3.06(m,2H),2.74(t,J＝6.3Hz,2H),2.68–2.59(m,2H),2.45–2.38(m,1H),2.32(s,3H),2.24(s,3H),2.13–1.99(m,2H),1.92–1.86(m,2H),1.84–1.63(m,4H).LC-MS:ESI m/z 531.3[M+H] +,C 30H 38N 6O 3.计算值530.30.
化合物233(甲酸盐)： 1H NMR(400MHz,CD 3OD)δ7.77–7.40(m,5H),6.49–6.46(m,1H),6.09(s,1H),4.81–4.76(m,1H),4.03–4.00(m,1H),3.79–3.68(m,1H),3.66–3.56(m,1H),3.48–3.39(m,4H),3.33–3.31(m,1H),3.29–3.22(m,1H),2.87–2.70(m,3H),2.68–2.60(m,1H),2.56–2.46(m,1H),2.32(s,3H),2.23(s,3H),2.26–2.06(m,2H),2.01–1.85(m,4H).LC-MS:ESI m/z 545.2[M+H] +；C 30H 36N 6O 4计算值544.28.
化合物234(甲酸盐)： 1H NMR(400MHz,CD 3OD)δ7.68–7.58(m,2H),7.58–7.49(m,2H),7.19(d,J＝7.3Hz,1H),6.36(d,J＝7.3Hz,1H),6.10(s,1H),4.27(s,2H),3.87(d,J＝11.0Hz,1H),3.79–3.72(m,1H),3.51–3.40(m,4H),3.27(d,J＝5.4Hz,1H),3.05–2.92(m,3H),2.88–2.82(m,1H),2.71(t,J＝6.2Hz,2H),2.37(dd,J＝13.3,6.5Hz,1H),2.32(s,3H),2.25(s,3H),2.22–2.15(m,1H),1.93–1.83(m,2H).LC-MS:ESI m/z 503.1[M+H] +；C 28H 34N 6O 3计算值502.27.
化合物235(甲酸盐)： 1H NMR(400MHz,CD 3OD)δ7.72–7.57(m,2H),7.56–7.46(m,2H),7.41(d,J＝7.3Hz,1H),6.50(d,J＝7.3Hz,1H),6.09(s,1H),4.41(d,J＝16.8Hz,1H),4.07(d,J＝16.7Hz,1H),3.68(dd,J＝11.7,4.1Hz,1H),3.55–3.40(m,4H),3.37–3.31(m,2H),3.21–3.10(m,2H),2.76(t,J＝6.1Hz,2H),2.70–2.59(m,2H),2.42–2.34(m,1H),2.31(s,3H),2.24(s,3H),2.09–2.00(m,1H),1.95–1.86(m,2H),1.82–1.65(m,4H),1.53–1.42(m,2H).LC-MS:ESI m/z 545.2[M+H] +；C 31H 40N 6O 3计算值544.32.
化合物236(甲酸盐)： 1H NMR(400MHz,CD 3OD)δ7.68–7.35(m,5H),6.55–6.49(m,1H),6.09(s,1H),4.59(d,J＝16.8Hz,1H),4.28(d,J＝7.9Hz,1H),4.01–3.57(m,4H),3.48–3.40(m,4H),3.33–3.32(m,1H),2.85–2.69(m,2H),2.30(s,3H),2.25(s,3H),2.21–1.73(m,4H).LC-MS:ESI m/z 517.1[M+H] +；C 28H 32N 6O 4计算值516.25.
化合物237(甲酸盐)： 1H NMR(400MHz,CD 3OD)δ7.66–7.35(m,5H),6.55–6.49(m,1H),6.08(s,1H),4.50(d,J＝16.5Hz,1H),4.25(s,1H),4.13–3.89(m,2H),3.75–3.54(m,2H),3.45–3.41(m,2H),3.34–3.32(m,1H),3.25–3.16(m,1H),2.95–2.87(m,2H),2.79–2.72(m,2H),2.66–2.57(m,1H),2.30(s,3H),2.23(s,3H),2.18–1.84(m,4H).LC-MS:ESI m/z 531.3[M+H] +；C 29H 34N 6O 4计算值530.26.
实施例38.化合物238的合成
Figure PCTCN2021078537-appb-000202
步骤1：(3-(3,5-二甲基-1H-吡唑-1-基)苯基)甘氨酸甲酯
Figure PCTCN2021078537-appb-000203
在N 2气氛下，向3-(3,5-二甲基-1H-吡唑-1-基)苯胺(1.8g，9.6mmol)的丙酮(20mL)溶液中加入K 2CO 3(1.9g，14mmol)。在60℃下搅拌1小时后，逐滴加入溴乙酸甲酯(1mL，15mmol)，并将混合物在60℃下搅拌2天。过滤混合物，并在真空下除去溶剂。残余物通过快速柱色谱法纯化(0-80％乙酸乙酯的石油醚溶液)，得到标题化合物(1.9g，76％)，为黄色油状物。 1H NMR(400MHz,DMSO-d 6)δ7.19–7.13(m,1H),6.64–6.63(m,2H),6.54(dd,J＝7.4,1.4Hz,1H),6.29(t,J＝6.4Hz,1H),6.01(s,1H),3.96(d,J＝6.4Hz,2H),3.65(s,3H),2.24(s,3H),2.15(s,3H).LC-MS:ESI m/z 260.24[M+H] +,C 14H 17N 3O 2计算值259.13.
步骤2：3-(((3-(3,5-二甲基-1H-吡唑-1-基)苯基)(2-甲氧基-2-氧乙基)氨基)甲基)吡咯烷-1-甲酸叔丁酯
Figure PCTCN2021078537-appb-000204
在N 2气氛下，向(3-(3,5-二甲基-1H-吡唑-1-基)苯基)甘氨酸甲酯(760mg，2.9mmol)和3-甲酰基吡咯烷-1-羧酸叔丁酯(696.5mg，3.5mmol)的甲醇(15mL)溶液中加入三乙酰氧基硼氢化钠(911.6mg，4.3mmol)。在25℃下搅拌16小时后，将氰基硼氢化钠(270.9mg，4.3mmol)加入到反应混合物中。在25℃下进一步搅拌16小时后，真空除去溶剂。将残余物用NaHCO 3(20mL)稀释，并用二氯甲烷(10mL×3)萃取。合并的有机相用盐水洗涤，经 硫酸钠干燥，并过滤。将滤液在减压下浓缩。残余物通过快速柱色谱法纯化(0-100％乙酸乙酯的石油醚溶液)，得到标题化合物(417mg，32％)，为浅黄色油状物。LC-MS:ESI m/z 443.4[M+H] +,C 24H 34N 4O 4计算值442.26.
步骤3-6参照化合物97的合成方法(步骤8-11)进行，制备得到化合物238(甲酸盐)：
1H NMR(400MHz,DMSO-d 6)δ7.47(d,J＝8.0Hz,1H),7.23(t,J＝8.1Hz,1H),7.11(d,J＝7.9Hz,1H),7.05(d,J＝7.2Hz,1H),6.70–6.64(m,2H),6.46(s,1H),6.28(d,J＝7.3Hz,1H),6.02(s,1H),4.06(s,2H),3.42(d,J＝7.3Hz,2H),3.25–3.22(m,2H),3.08–3.02(m,2H),2.93–2.85(m,2H),2.70(t,J＝7.4Hz,2H),2.60(t,J＝6.1Hz,2H),2.29(s,2H),2.25(s,3H),2.15(s,3H),2.05–1.99(m,2H),1.77–1.71(m,2H),1.66–1.60(m,1H).LC-MS:ESI m/z 489.3[M+H] +,C 28H 36N 6O 2计算值488.29.
实施例39.参照化合物238的合成方法，制备得到化合物239(甲酸盐)：
1H NMR(400MHz,DMSO-d 6)δ7.20(t,J＝8.1Hz,1H),7.02(d,J＝7.2Hz,1H),6.70–6.57(m,3H),6.43(brs,1H),6.23(d,J＝7.2Hz,1H),6.00(s,1H),3.95(s,2H),3.37(d,J＝6.9Hz,2H),3.25–3.20(m,2H),2.96–2.90(m,1H),2.80–2.75(m,1H),2.71–2.54(m,7H),2.46–2.40(m,2H),2.23(s,3H),2.15(s,3H),2.00–1.93(m,1H),1.81–1.68(m,4H),1.58–1.50(m,1H).LC-MS:ESI m/z 503.4[M+H] +；C 29H 38N 6O 2计算值502.31.
实施例40.化合物240的合成
Figure PCTCN2021078537-appb-000205
步骤1：3-((3-(3,5-二甲基-1H-吡唑-1-基)苯基)(3-乙氧基-3-氧丙基)氨基甲酰基)吡咯烷-1-甲酸叔丁酯
Figure PCTCN2021078537-appb-000206
在N 2气氛下，向3-((3-(3,5-二甲基-1H-吡唑-1-基)苯基)氨基甲酰基)吡咯烷-1-甲酸叔丁酯(650mg，1.7mmol)的原硅酸四乙酯(1.5mL)溶液中加入CsF(236mg，1.7mmol)和丙烯酸乙酯(0.25mL，2.5mmol)。在50℃下搅拌16小时后，用水(10mL)将反应淬灭，并用乙酸乙酯(8mL×3)萃取，将合并的有机相用盐水洗涤，用硫酸钠干燥，并过滤。将滤液在减压下浓缩。残余物通过快速柱色谱法纯化(0-80％乙酸乙酯的石油醚溶液)，得到标题化合物(89mg，29％)，为无色油状物。LC-MS:ESI m/z 485.4[M+H] +；C 26H 36N 4O 5计算值484.27.
步骤2：3-(((3-(3,5-二甲基-1H-吡唑-1-基)苯基)(2-甲氧基-2-氧乙基)氨基)甲基)吡咯烷-1-甲酸叔丁酯
Figure PCTCN2021078537-appb-000207
向3-((3-(3,5-二甲基-1H-吡唑-1-基)苯基)氨基甲酰基)吡咯烷-1-甲酸叔丁酯(89mg，0.2mmol)的二氯甲烷(1mL)溶液中加入盐酸/1，4-二氧六环(1mL)。在25℃下搅拌3小时后，真空除去溶剂，得到标题化合物(85mg，粗品)，为无色油状物，其无需进一步纯化直接用于下一步。LC-MS:ESI m/z 385.3[M+H] +；C 21H 28N 4O 3计算值384.22.
步骤3-5参照化合物97的合成方法(步骤9-11)，制备得到化合物240(甲酸盐)：
1H NMR(400MHz,DMSO-d 6)δ7.60–7.52(m,2H),7.48(s,1H),7.31(d,J＝7.3Hz,1H),6.99(d,J＝7.3Hz,1H),6.28(s,1H),6.22(d,J＝7.2Hz,1H),6.09(s,1H),3.88–3.83(m,2H),3.24–3.19(m,4H),2.86–2.69(m,2H),2.65–2.54(m,7H),2.42(t,J＝7.5Hz,2H),2.33(s,3H),2.18(s,3H),1.95–1.88(m,1H),1.76–1.70(m,2H),1.63–1.54(m,1H).LC-MS:ESI m/z 517.33[M+H] +；C 29H 36N 6O 3计算值516.28.
实施例41.化合物241的合成
Figure PCTCN2021078537-appb-000208
步骤1：(S)-3-((2-甲氧基苯基)氨基甲酰基)吡咯烷-1-甲酸叔丁酯
Figure PCTCN2021078537-appb-000209
在0℃下，向2-甲氧基苯胺(1.0g，8.1mmol)，(S)-1-(叔丁氧羰基)吡咯烷-3-羧酸(2.1g，9.7mmol)，HOBT(1.6g，12mmol)和EDCI(2.3g，12mmol)的二氯甲烷(30mL)溶液中，缓慢滴加DIPEA(2.1g，16mmol)。将反应混合物在0℃下搅拌3小时。然后将反应混合物倒入冰水(50mL)中，将有机相用饱和NaHCO 3水溶液(15mL×2)洗涤，然后用盐水(15mL)洗涤，用无水硫酸钠干燥并浓缩至干。将残余物通过FCC纯化(20％-50％乙酸乙酯的石油醚溶液)，得到(S)-3-((2-甲氧基苯基)氨基甲酰基)氨基吡咯烷-1-羧酸 叔丁酯(1.8g，67.4％)，为粉红色固体。 1H NMR(400MHz,DMSO-d 6)δ9.25(brs,1H),7.90(dd,J＝8.0,1.2Hz,1H),7.11–7.02(m,2H),6.92–6.86(m,1H),3.83(s,3H),3.50(t,J＝8.4Hz,1H),3.42–3.34(m,2H),3.25–3.16(m,2H),2.15–1.98(m,2H),1.40(s,9H).LC-MS:ESI m/z321.4[M+H] +；C 17H 24N 2O 4计算值320.39.
步骤2：(S)-3-((2-甲氧基-2-氧乙基)(2-甲氧基苯基)氨基甲酰基)吡咯烷-1-甲酸叔丁酯
Figure PCTCN2021078537-appb-000210
向(S)-3-((2-甲氧基苯基)氨基甲酰基)吡咯烷-1-羧酸叔丁酯(0.8g，2.5mmol)的四氢呋喃(20mL)溶液中加入氢化钠(0.20g，5.0mmol)。将悬浮液在0℃下搅拌30分钟，然后缓慢加入2-溴乙酸甲酯(0.76g，5.0mmol)。将反应混合物在0℃下搅拌2小时。用冰水(20mL)淬灭反应溶液，并用二氯甲烷(10mL×3)萃取。有机相用盐水(10mL)洗涤，用无水硫酸钠干燥并浓缩至干。残余物通过FCC纯化(35％乙酸乙酯的石油醚溶液)，得到(S)-3-((2-甲氧基-2-氧乙基)(2-甲氧基苯基)氨基甲酰基)吡咯烷-1-甲酸叔丁酯(0.72g，73.3％)黄色油状物。 1H NMR(400MHz,DMSO-d 6)δ7.44–7.34(m,2H),7.18(d,J＝8.4Hz,1H),7.03(t,J＝7.6Hz,1H),4.72–4.64(m,1H),3.83(d,J＝1.8Hz,3H),3.80–3.74(m,1H),3.65(t,J＝7.6Hz,3H),3.30(s,2H),3.27–3.14(m,2H),3.10–3.00(m,2H),2.98–2.80(m,1H),1.38(s,9H).LC-MS:ESI m/z 393.5[M+H] +；C 20H 28N 2O 6计算值392.45.
步骤3：(S)-N-(2-甲氧基苯基)-N-(吡咯烷-3-羰基)甘氨酸甲酯
Figure PCTCN2021078537-appb-000211
向(S)-3-((2-甲氧基-2-氧乙基)(2-甲氧基苯基)氨基甲酰基)吡咯烷-1-羧酸叔丁酯(0.30g，0.76mmol)的甲醇(1mL)溶液中加入盐酸/1，4-二氧六环(4N，1.5mL)。将反应混合物在25℃下搅拌16小时。浓缩反应混合物，得到黄色油状的(S)-N-(2-甲氧基苯基)-N-(吡咯烷-3-羰基)甘氨酸甲酯(0.23g，粗品)。 1H NMR(400MHz,DMSO-d 6)δ9.08(brs,2H),7.46–7.38(m,2H),7.22–7.18(m,1H),7.08–7.02(m,1H),4.70–4.63(m,1H),3.84(s,3H),3.70–3.60(m,3H),3.26–2.99(m,6H),2.03–1.80(m,2H).LC-MS:ESI m/z 293.3[M+H] +；C 15H 20N 2O 4计算值292.34.
步骤4：(S)-7-(3-(3-((2-甲氧基-2-氧代乙基)(2-甲氧基苯基)氨基甲酰基)吡咯烷-1-基)丙基)-3,4-二氢-1,8-萘啶-1(2H)-羧酸叔丁酯
Figure PCTCN2021078537-appb-000212
向(S)-N-(2-甲氧基苯基)-N-(吡咯烷-3-羰基)甘氨酸甲酯(0.23g，0.79mmol)，7-(3- 氧丙基)-3,4-二氢-1,8-萘啶-1(2H)-羧酸叔丁酯(0.23g，0.79mmol)和三乙酰氧基硼氢化钠(0.34g，1.6mmol)的DCE(5mL)溶液中滴加DIPEA(0.51g，4.0mmol)。将反应混合物在25℃下搅拌16小时。将反应混合物倒入冰水(10mL)中，并用二氯甲烷(5mL×3)萃取。合并的有机相用盐水(5mL)洗涤，经无水硫酸钠干燥并浓缩至干。残余物通过FCC纯化(5％甲醇的二氯甲烷溶液)，得到(S)-7-(3-(3-((2-甲氧基-2-氧代乙基)(2-甲氧基苯基)氨基甲酰基)吡咯烷-1-基)丙基)-3,4-二氢-1,8-萘啶-1(2H)-羧酸叔丁酯(0.11g，25.3％)，为无色油状物。 1H NMR(400MHz,DMSO-d 6)δ7.43–7.32(m,3H),7.15(d,J＝7.5Hz,1H),7.04–6.99(m,1H),6.85(dd,J＝7.6,3.6Hz,1H),4.70–4.64(m,1H),3.80(s,3H),3.76–3.70(m,1H),3.63(s,3H),3.60(d,J＝5.2Hz,2H),2.76–2.72(m,1H),2.67(t,J＝6.6Hz,2H),2.62–2.52(m,4H),2.49–2.20(m,6H),1.82–1.77(m,2H),1.77–1.68(m,2H),1.45–1.35(m,9H).LC-MS:ESI m/z 567.7[M+H] +；C 31H 42N 4O 6计算值566.70.
步骤5：(S)-N-(2-甲氧基苯基)-N-(1-(3-(5,6,7,8-四氢-1,8-萘啶-2-基)丙基)吡咯烷-3-羰基)甘氨酸甲酯
Figure PCTCN2021078537-appb-000213
向(S)-7-(3-(3-((2-甲氧基-2-氧乙基)(2-甲氧基苯基)氨基甲酰基)吡咯烷-1-基)丙基)-3,4-二氢-1,8-萘啶-1(2H)-羧酸叔丁酯(0.11g，0.20mmol)的甲醇(1.5mL)溶液中加入盐酸/1，4-二氧六环(4N，2mL)。将反应混合物在25℃下搅拌16小时。浓缩反应液，得到(S)-N-(2-甲氧基苯基)-N-(1-(3-(5,6,7,8-四氢-1,8-萘啶-2-基)丙基)吡咯烷-3-羰基)甘氨酸甲酯(0.12g，粗品)，为黄色油状物。 1H NMR(400MHz,DMSO-d 6)δ8.07–8.00(m,1H),7.62(d,J＝7.2Hz,1H),7.46–7.37(m,2H),7.20(d,J＝8.4Hz,1H),7.05(dd,J＝8.8,6.4Hz,1H),6.72–6.60(m,1H),4.72–4.61(m,2H),3.91–3.81(m,4H),3.65(d,J＝7.2Hz,3H),3.53–3.47(m,4H),3.21–3.15(m,2H),3.09–3.00(m,2H),2.77–2.69(m,4H),2.19–1.93(m,4H),1.82(d,J＝4.8Hz,2H).LC-MS:ESI m/z 467.6[M+H] +；C 26H 34N 4O 4计算值466.58.
步骤6：(S)-N-(2-甲氧基苯基)-N-(1-(3-(5,6,7,8-四氢-1,8-萘啶-2-基)丙基)吡咯烷-3-羰基)甘氨酸
Figure PCTCN2021078537-appb-000214
向(S)-N-(2-甲氧基苯基)-N-(1-(3-(5,6,7,8-四氢-1,8-萘啶-2-基)丙基)吡咯烷-3-羰基)甘氨酸甲酯(0.12g，0.26mmol)的甲醇(1mL)溶液中加入氢氧化锂水溶液(2N，1.0mL)。将反应混合物在25℃下搅拌16小时。浓缩反应混合物以除去甲醇。加入盐酸(1N)调节pH＝6-7，将反应物冻干并通过制备HPLC纯化(色谱柱：Kromasil-C18 100×21.2mm 5um；流动相：乙腈-水(0.1％甲酸)；梯度：5-15％)得到(S)-N-(2-甲氧基苯基)-N-(1-(3-(5,6,7,8- 四氢-1,8-萘啶-2-基)丙基)吡咯烷-3-羰基)甘氨酸(78mg，66％)，为黄色固体。 1H NMR(400MHz,DMSO-d 6)δ7.45–7.35(m,2H),7.15(dd,J＝7.6,4.4Hz,1H),7.06–7.00(m,2H),6.65(brs,1H),6.24–6.21(m,1H),4.66–4.62(m,1H),3.81(d,J＝0.8Hz,3H),3.47–3.42(m,1H),3.23(d,J＝4.4Hz,2H),2.84–2.70(m,2H),2.65–2.53(m,4H),2.45–2.32(m,5H),1.92–1.84(m,1H),1.76–1.62(m,4H),1.53–1.45(m,1H).LC-MS:ESI m/z 453.3[M+H] +；C 25H 32N 4O 4计算值452.26.HPLC纯度:100.0％(214nm),100.0％(254nm).实施例42.参照化合物241的合成，制备得到以下化合物：
化合物242(甲酸盐)： 1H NMR(400MHz,DMSO-d 6,80℃)δ7.48–7.38(m,5H),7.06(d,J＝7.2Hz,2H),6.68(brs,1H),6.26(d,J＝7.2Hz,1H),4.17–4.15(m,2H),3.25–3.23(m,2H),2.98–2.94(m,1H),2.81–2.70(m,3H),2.62–2.57(m,3H),2.46–2.39(m,3H),1.97–1.94(m,1H),1.78–1.63(m,5H).LC-MS:(ESI)m/z 423.3[M+H] +,C 24H 30N 4O 3计算值422.23.HPLC纯度:99.7％(254nm),99.6％(214nm).
化合物243(铵盐)： 1H NMR(400MHz,DMSO-d 6)δ7.48–7.35(m,4H),7.12(d,J＝7.2Hz,1H),6.27(d,J＝7.2Hz,1H),4.20–4.12(m,1H),4.07–4.00(m,1H),3.27–3.20(m,2H),3.00–2.85(m,1H),2.69–2.58(m,3H),2.48–2.12(m,7H),2.05–2.00(m,1H),1.79–1.70(m,2H),1.58–1.55(m,2H),1.39–1.35(m,3H).LC-MS:(ESI)m/z 471.3,473.3[M+H,Cl] +；C 25H 31ClN 4O 3计算值470.21.HPLC纯度:100.0％(214nm),100.0％(254nm).
化合物244(甲酸盐)： 1H NMR(400MHz,DMSO-d 6,80℃)δ7.46–7.40(m,2H),7.35–7.32(m,3H),7.04(d,J＝7.2Hz,1H),6.58(brs,1H),6.24(d,J＝7.2Hz,1H),4.24(d,J＝17.2Hz,1H),4.11(d,J＝17.2Hz,1H),3.29–3.22(m,2H),2.92(s,1H),2.67–2.49(m,5H),2.38–2.31(m,5H),1.97–1.94(m,1H),1.79–1.72(m,2H),1.67–1.56(m,2H),1.45-1.37(m,3H).LC-MS:(ESI)m/z 437.3[M+H] +；C 25H 32N 4O 3计算值436.25.HPLC纯度:97.9％(214nm),98.9％(254nm).
化合物245(甲酸盐)： 1H NMR(400MHz,DMSO-d 6)δ7.47(brs,1H),7.43–7.38(m,5H),7.14(d,J＝7.2Hz,1H),6.30(d,J＝7.2Hz,1H),4.32–4.24(m,1H),4.04–3.95(m,1H),3.30–3.25(m,2H),3.01(m,1H),2.86(m,1H),2.53–2.43(m,5H),2.52–2.43(m,4H),2.06–2.03(m,1H),1.83–1.64(m,4H),1.43(brs,3H).LC-MS:(ESI)m/z 437.3[M+H] +；C 25H 32N 4O 3计算值436.25.HPLC纯度:96.6％(214nm),95.7％(254nm).
化合物246(甲酸盐)： 1H NMR(400MHz,DMSO-d 6)δ8.05(brs,1H),7.50(q,J＝8.8Hz,4H),7.16(d,J＝7.2Hz,1H),6.30(d,J＝7.2Hz,1H),4.31–4.16(m,1H),3.99–3.88(m,1H),3.31–3.24(m,2H),3.02–2.97(m,1H),2.92–2.75(s,1H),2.60–2.52(m,5H),2.46–2.41(m,3H),2.39–2.30(m,1H),2.14–2.01(m,1H),1.77–1.73(m,2H),1.72–1.60(m,2H),1.48–1.33(m,3H).LC-MS:ESI m/z 471.2,473.2[M+H,Cl] +；C 25H 31ClN 4O 3计算值470.21.HPLC纯度:97.9％(214nm),97.9％(254nm).
化合物247(甲酸盐)： 1H NMR(400MHz,DMSO-d 6)δ8.20(brs,1H),7.43–7.36(m,2H),7.18–7.13(m,1H),7.08–6.99(m,2H),6.26(t,J＝7.2Hz,1H),4.67–4.60(m,1H),3.81(d,J＝1.6Hz,3H),3.60–3.40(m,1H),3.25–3.20(m,2H),2.92–2.78(m,4H),2.69–2.67(m,2H),2.61–2.59(m,3H),2.46–2.41(m,2H),1.97–1.85(m,1H),1.78–1.71(m,4H),1.65–1.51(m,1H).LC-MS:(ESI)m/z 453.3[M+H] +,C 25H 32N 4O 4计算值452.24.HPLC纯度:99.4％(254nm),97.3％(214nm).
化合物248(甲酸盐)： 1H NMR(400MHz,DMSO-d 6)δ8.18(brs,1H),7.70–7.61(m,2H),7.50–7.32(m,2H),7.12–7.00(m,1H),6.35–6.21(m,1H),4.68–4.60(m,1H),3.58–3.52(m,1H), 3.27–3.21(m,2H),2.72–2.68(m,2H),2.63–2.58(m,4H),2.49–2.32(m,5H),1.99–1.91(m,1H),1.80–1.57(m,5H).LC-MS:(ESI)m/z 457.2,459.2[M+H,Cl] +；C 24H 29ClN 4O 3计算值456.19.HPLC纯度:98.9％(254nm),99.1％(214nm).
实施例43 化合物249的合成
Figure PCTCN2021078537-appb-000215
步骤1:3-((3-(3,5-二甲基-1H-吡唑-1-基)苯基)氨基)丙酸甲酯
Figure PCTCN2021078537-appb-000216
在N 2气氛下，向3-(3,5-二甲基-1H-吡唑-1-基)苯胺(1.0g，5.3mmol)和3-溴丙酸甲酯(0.7mL，6.3mmol)的甲醇(5mL)溶液中添加三乙胺(5mL)。在80℃下搅拌1小时后，真空除去溶剂，并通过快速柱色谱法纯化(0-80％乙酸乙酯的石油醚溶液)，得到标题化合物(537mg，38％)，为黄色油状物。LC-MS:ESI m/z 274.36[M+H] +,C 15H 19N 3O 2计算值273.15.
步骤2：3-(((3-(3,5-二甲基-1H-吡唑-1-基)苯基)(3-甲氧基-3-氧丙基)氨基)甲基)吡咯烷-1-甲酸叔丁酯
Figure PCTCN2021078537-appb-000217
在N 2气氛下，向3-((3-(3,5-二甲基-1H-吡唑-1-基)苯基)氨基)丙酸甲酯(537mg，1.9mmol)和3-甲酰基吡咯烷-1-羧酸叔丁酯(498mg，2.5mmol)的甲醇(10mL)，冰醋酸(75mg，1.3mmol)溶液中加入三乙酰氧基硼氢化钠(604.2mg，2.8mmol)。在25℃下搅拌16小时后，将氰基硼氢化钠(176.4mg，2.8mmol)加入到反应混合物中。在25℃下进一步搅拌16小时后，真空除去溶剂。将残余物用NaHCO 3(20mL)稀释并用二氯甲烷(10mL×3)萃取。合并的有机相用盐水洗涤，经硫酸钠干燥，并过滤。将滤液在减压下浓缩。残余物通过快速柱色谱法纯化(0-100％乙酸乙酯的石油醚溶液)，得到标题化合物(402mg，75％)，为黄色油状物。LC-MS:ESI m/z 457.45[M+H] +,C 25H 36N 4O 4计算值456.27.
步骤3：3-((3-(3,5-二甲基-1H-吡唑-1-基)苯基)(吡咯烷-3-基甲基)氨基)丙酸甲酯
Figure PCTCN2021078537-appb-000218
向3-(((3-(3,5-二甲基-1H-吡唑-1-基)苯基)(3-甲氧基-3-氧丙基)氨基)甲基)吡咯烷-1-甲酸叔丁酯(284mg，0.6mmol)的二氯甲烷(3mL)溶液中加入盐酸/1，4-二氧六环(1mL)。在25℃搅拌2小时后，真空除去溶剂，得到盐酸盐形式的标题化合物(212mg，99％)，为黄色固体，其无需进一步纯化直接用于下一步。LC-MS:ESI m/z 357.32[M+H] +,C 20H 28N 4O 2计算值356.22.
步骤4：7-(2-(3-(((3-(3,5-二甲基-1H-吡唑-1-基)苯基)(3-甲氧基-3-氧丙基)氨基)甲基)吡咯烷丁酸叔丁基-1-基)乙基)-3,4-二氢-1,8-萘啶-1(2H)-羧酸叔丁酯
Figure PCTCN2021078537-appb-000219
在N 2气氛下，向盐酸盐形式的3-((3-(3,5-二甲基-1H-吡唑-1-基)苯基)(吡咯烷-3-基甲基)氨基)丙酸甲酯(212mg，0.6mmol)和7-(2-碘乙基)-3,4-二氢-1,8-萘啶-1(2H)-羧酸叔丁酯(359mg，0.9mmol)的乙腈(10mL)溶液中加入K 2CO 3(414mg，3mmol)。在25℃下搅拌16小时后，真空除去溶剂，并将粗残余物通过快速柱色谱纯化(0～6％甲醇的二氯甲烷溶液)得到标题化合物(353mg，95％)，为褐色固体。LC-MS:ESI m/z 617.50[M+H] +,C 35H 48N 6O 4计算值616.37.
步骤5：3-(((1-(2-(8-(叔丁氧羰基)-5,6,7,8-四氢-1,8-萘啶-2-基)乙基)吡咯烷-3-基)甲基)(3-(3,5-二甲基-1H-吡唑-1-基)苯基)氨基)丙酸
Figure PCTCN2021078537-appb-000220
向7-(2-(3-(((3-(3,5-二甲基-1H-吡唑-1-基)苯基)(3-甲氧基-3-氧丙基)氨基)甲基)吡咯烷-1-))-3,4-二氢-1,8-萘啶-1(2H)-羧酸叔丁酯(155mg，0.25mmol)在甲醇(2mL)和H 2O(0.5mL)的混合溶液中添加LiOH(7mg，0.27mmol)。在25℃下搅拌2小时后，真空除去溶剂，得到标题化合物(150mg，粗品)，为黄色固体锂盐形式，其无需进一步纯化即可用于下一步。LC-MS:ESI m/z 603.4[M+H] +,C 34H 46N 6O 4计算值602.36
步骤6：3-((3-(3,5-二甲基-1H-吡唑-1-基)苯基)((1-(2-(5,6,7,8-四氢-1,8-萘啶-2-基)乙基)吡咯烷-3-基)甲基)氨基)丙酸
Figure PCTCN2021078537-appb-000221
向锂盐(150mg，0.25mmol)形式的3-(((1-(2-(8-(叔丁氧羰基)-5,6,7,8-四氢-1,8-萘啶-2-基)乙基)吡咯烷-3-基)甲基)(3-(3,5-二甲基-1H-吡唑-1-基)苯基)氨基)丙酸在二氯甲烷(1mL)的溶液中加入盐酸/1，4-二氧六环(4N，1mL)。在25℃下搅拌3小时后，在真空下除去溶剂，并将粗制残留物通过制备型反相HPLC在以下条件下纯化[色谱柱：Kromasil100-5-C18，30×150mm；流动相：1-100％乙腈(含0.1％的甲酸)，30分钟]，得到浅黄色固体状的甲酸盐形式的化合物249(39.1mg，34％)。 1H NMR(400MHz,DMSO-d 6)δ7.28–7.19(m,1H),7.02(d,J＝7.3Hz,1H),6.74–6.72(m,2H),6.67(d,J＝8.0Hz,1H),6.33(s,1H),6.26(d,J＝7.3Hz,1H),6.02(s,1H),3.65–3.55(m,2H),3.32–3.26(m,2H),3.25–3.21(m,2H),2.84–2.72(m,3H),2.63–2.52(m,8H),2.45(t,J＝7.1Hz,2H),2.27(s,3H),2.15(s,3H),1.95–1.85(m,1H),1.78–1.70(m,2H),1.50–1.40(m,1H).LC-MS:ESI m/z 503.3[M+H] +；C 29H 38N 6O 2计算值502.31.
生物活性评价实验
实验材料：带有His标签的人类整合素蛋白购自Acro Biosystems公司，包括α5β1(IT1-H52W5)，α8β1(IT1-H52W9)，αvβ1(IT1-H52E1)，αvβ3(IT3-H52E3)，αvβ5(IT8-H52W5)，αvβ6(IT6-H52E1)，αvβ8(IT8-H52W4)。蛋白用无菌水溶解，分装后储存于-80℃。生物素标记的多肽由GenScript合成，DMSO溶解后分装存于-20℃。
Figure PCTCN2021078537-appb-000222
Hisdidine Detection Kit(Nickel Chelate)(6760619M)购自于PerkinElmer公司。Source板(PP-0200)购自于Labcyte公司，Proxiplate-384plus目标板购自于Perkin Elmer公司。反应缓冲液自行配制：25mM Tris pH7.4，150mM NaCl，0.1％BSA，1mM MgCl 2，1mM CaCl 2。
实验方法：将备选化合物用DMSO溶解，按一定浓度梯度加到Source板上，然后用ECHO550将化合物转移到目标板，使之形成11个浓度。按下表所示的整合素对应RGD多肽以及工作浓度，用反应缓冲液稀释，轻柔混匀，并用Multidrop以每孔10μl加到目标板中，室温孵育1个小时。每孔加入10μl Donor/Acceptor beads(终浓度15μg/mL)，室温孵育一个小时。最后用Envision Plate Reader读数。每个化合物的IC 50用XLfit拟合后获得。下表1为本发明化合物对不同整合素亚型抑制活性的效果。
Figure PCTCN2021078537-appb-000223
表1：本发明化合物对不同整合素亚型抑制活性的效果评价
注：A：<100nM；B：100-1000nM；C：>1000nM
  α5β1 α8β1 αvβ1 αvβ3 αvβ5 αvβ6 αvβ8
化合物1   C C C C C C
化合物2   C C C C C C
化合物3   B C C C C B
化合物4   C C C C C C
化合物5a C C C B B C C
化合物5b C B B A A A A
化合物6   C C A A B B
化合物7   C C B B C B
化合物8   B C B B B A
化合物9   C C C C C C
化合物10a C C C C C C C
化合物10b C C C A A B B
化合物11a C C C C C C B
化合物11b C A B B B A A
化合物12a C C C C C C C
化合物12b C C C B C C B
化合物13a C C C C C C C
化合物13b C B C B B B A
化合物14a C C C C C C C
化合物14b C C C B B B B
化合物15   C C B C C C
化合物16a   C C C C C C
化合物16b   C C C C C C
化合物17   B C B B C C
化合物18   C C B C C C
化合物19   C C C C C C
化合物20   C C C C C C
化合物21   B C C C C C
化合物22   B C C C C C
化合物23 B A B C B B C
化合物24a C B C C C C C
化合物24b B A C C B B C
化合物26 B A B C B B C
化合物27a A A A B B B B
化合物27b B B B C C C C
化合物28a A A B B A A B
化合物28b C C C C C C C
化合物29   A A C B B B
化合物29a A A A C B B B
化合物29b C B C C C C C
化合物30   B C C C C C
化合物31 B A B C B B C
化合物32   A B C B B C
化合物33   A A C B A B
化合物33a A A A C B B B
化合物33b C B C C C C C
化合物34 A A B C A B B
化合物35 B A B C B B C
化合物36 B A B C B B C
化合物37 B A B B B B C
化合物38 A A B B B B C
化合物39 B A B B B B B
化合物39a A A B B A A B
化合物39b C C C C C C C
化合物40 B A B B A B C
化合物42 B A B B B B B
化合物43 B A B C B B C
化合物45 B A B C B B C
化合物46a A A A C B A B
化合物46b   A C C C C C
化合物49a B A B C C C C
化合物49b C B C C C C C
化合物51a C B C C C C C
化合物51b B A B C C B B
化合物55a A A A A A B C
化合物55b C B C C C C C
化合物57 B A B A A B B
化合物58 A A A A A B B
化合物63 A A A C C A B
化合物65 A A B C B A B
化合物65a C B C C C B C
化合物65b A A A B B A B
化合物68 A A A C C B B
化合物70a A A A B B B B
化合物70b C C C C C C C
化合物72 B A B C C B C
化合物73 A A B C C B B
化合物74 A A B B B B C
化合物77 B A B B B B C
化合物78 A A B B B B C
化合物81 B A B A A B C
化合物82 B A B A A B C
化合物86 B A B C C A B
化合物91a A A A C C A A
化合物91b C B C C C C C
化合物95 A A B B B B B
化合物96 A A A A A A B
化合物97   A A C C A A
化合物98   A B C C B C
化合物99a C A C C C C C
化合物99b B A B C C B B
化合物100   B C C C C C
化合物101 A A B C C B B
化合物102   A B C C C B
化合物103 C A C C C B B
化合物104   A B C C C C
化合物105a A A A C C A A
化合物105b C B C C C C C
化合物106   A B C C B C
化合物107   A C C C C C
化合物108   C C C C C C
化合物109   B C B B B C
化合物110   A B B C B C
化合物111   A B C C B C
化合物112 C B C C C C C
化合物113 C B B C C C C
化合物114   C C C C C C
化合物115   C C C C C C
化合物116   C C C C C C
化合物117   C C C C C C
化合物121   C C C C C C
化合物122   C C C C C C
化合物125   B C B A C B
化合物126   C C C C C C
化合物127   C C C C C C
化合物128   C C C C C C
化合物129   C C C B C C
化合物130   B C C C C C
化合物131   B C C C C C
化合物134   C C B B C C
化合物134a C C C C C C C
化合物134b C C C A A C C
化合物134c   C C B B C C
化合物135   C C C B C C
化合物136   B C B B C B
化合物138   B C B B C B
化合物139   A B A B B B
化合物140   B C C C C C
化合物141   C C C C C C
化合物142   C C C C C C
化合物143   B C C C C C
化合物144   C C C C C C
化合物145   C C C C C C
化合物146 C B B C C C A
化合物147 B A A C C C A
化合物149 C A B B B A A
化合物150 C B B B B B A
化合物151 C B B A A A A
化合物152 C B C A B B A
化合物154 C B C A B A A
化合物155 C C C A B A A
化合物156 C B B B B A A
化合物157 C B B A A A A
化合物158 C B B A A A A
化合物158a C C C A B B B
化合物158b C C B A A A A
化合物159 C A B C C B A
化合物159a B B A B C A A
化合物159b C C C C C C B
化合物160 C B B B C A A
化合物161 C A B B C A A
化合物162 C A B C C A A
化合物166 C B C B C A A
化合物166a C C C C C C B
化合物166b C B B B C A A
化合物167 C A B A B A A
化合物168 C A B B C B A
化合物172 C A A C C A A
化合物173 C A B B B A A
化合物177 C B B B B A A
化合物177a C C C C C B B
化合物177b C B B A B A A
化合物178 C B B B C A A
化合物182 C A B B B A A
化合物184 C B B B B B A
化合物186 C B B A B A A
化合物188 C B B A B A A
化合物189 C B B B B B A
化合物190 C C C B C B A
化合物193 C C C B C C B
化合物194 C B B A B A A
化合物195 C A B B C A A
化合物195a C C C C C C B
化合物195b C B B A C A A
化合物196 B A A C C B A
化合物200 C A B C C B A
化合物201 C C B B B B A
化合物202 C B B B C B A
化合物203 C B B C C A A
化合物204 C B B C C A A
化合物205 B A A C C A A
化合物207 B B A C C A A
化合物208 C B B C C A A
化合物211 C B B C C A A
化合物212 C C C C C C A
化合物213 C B B C C A A
化合物214 C B B C C A A
化合物215 C B B C C A A
化合物216 C B B C C C A
化合物217 C C C C C A B
化合物221 C B B B C A A
化合物224 C B B A C B A
化合物225 C B B B C C B
化合物226 C B B A B B A
化合物227 C B B A B A A
化合物228 C B B C C A A
化合物229 C B A C C A A
化合物230   B C C C C C
化合物230a   C C C C C C
化合物230b   B C C C C C
化合物231   A C C C B B
化合物231a   A C C C B B
化合物231b   B C C C B B
化合物232   B C C C B B
化合物232a   B C C C C C
化合物232b   B C C C B B
化合物233   B C C C C B
化合物234   B C C C C C
化合物235   B C C C C B
化合物236   C C C C C C
化合物237   A C C C C B
化合物238   C C C C C C
化合物239   B C C C C C
化合物240   A B C C C C
化合物241   A C C C C C
化合物242   A C C C C B
化合物243   B C C C C B
化合物244   B C C C C C
化合物245   B C C C C C
化合物246   B C C C C C
化合物247   B C C C C C
化合物248   A C C C C C
化合物249   A B C C C C
上述实验结果表明：本发明化合物对不同整合素亚型均具有良好抑制作用。
以上对本发明的实施方式进行了说明。但是，本发明不限定于上述实施方式。凡在本发明的精神和原则之内，所做的任何修改、等同替换、改进等，均应包含在本发明的保护范围之内。

Claims (16)

  1. 一种式I所示的化合物及其消旋体、立体异构体、互变异构体、同位素标记物、氮氧化物、溶剂化物、多晶型物、代谢产物、酯、前药或其药学上可接受的盐：
    Figure PCTCN2021078537-appb-100001
    其中，所述R 1、R 5各自独立的选自H，OH，卤素，CN，SH，NH 2，COOH，任选被一个、两个或更多个R a取代的如下基团：C 1-C 12脂肪烃基，C 1-C 12脂肪烃基氧基；所述R a选自H，＝O，卤素，OH，CN，SH，NH 2，COOH；每一个R 1可以相同或不同；每一个R 5可以相同或不同；
    所述W选自-O-，-NR 2-，-S-；
    所述R 2、R 3各自独立的选自H，C 1-C 12脂肪烃基，C 3-12环烷基，C 3-12环烷基-C 1-C 12脂肪烃基，-L 5-Ar，并且R 2、R 3中至少一个选自-L 5-Ar；
    所述Ar选自任选被一个、两个或更多个R b取代的如下基团：C 3-12环烷基，3-12元杂环基，C 6-20芳基或5-14元杂芳基；所述R b选自H，＝O，卤素，OH，CN，SH，NH 2，COOH，或任选被一个、两个或更多个R c取代的如下基团：C 1-C 12脂肪烃基，C 1-C 12脂肪烃基氧基，C 1-C 12脂肪烃基-SO 2-，C 1-C 12脂肪烃基-NH-，二(C 1-C 12脂肪烃基)N-，C 3-12环烷基，C 3-12环烷基氧基，C 3-12环烷基-SO 2-，C 3-12环烷基-NH-，C 3-12环烷基-C 1-C 12脂肪烃基氧基，C 3-12环烷基-C 1-C 12脂肪烃基-SO 2-，C 3-12环烷基-C 1-C 12脂肪烃基-NH-，3-12元杂环基，3-12元杂环基氧基，3-12元杂环基-SO 2-，3-12元杂环基-NH-，C 6-20芳基，C 6-20芳基氧基，C 6-20芳基-SO 2-，C 6-20芳基-NH-，5-14元杂芳基，5-14元杂芳基氧基，5-14元杂芳基-SO 2-，5-14元杂芳基-NH-；所述R c选自H，＝O，卤素，OH，CN，SH，NH 2，COOH，C 1-C 12脂肪烃基，C 1-C 12脂肪烃基氧基，C 1-C 12脂肪烃基-SO 2-，C 1-C 12脂肪烃基-NH-，二(C 1-C 12脂肪烃基)N-；
    所述R 4独立的选自H，C 1-C 12脂肪烃基；
    所述L 1、L 2、L 3、L 4、L 5各自独立的选自-(CH 2) n-C(＝X)-或-C(＝X)-(CH 2) n-，-(CH 2) m-(当m＝0时，代表键)，-(CH 2) n-C(＝O)-NH-，-(CH 2) n-NH-C(＝O)-，-C(＝O)-NH-(CH 2) n-，-NH-C(＝O)-(CH 2) n-，-(CH 2) n-C(＝O)-NH-(CH 2) n-，-(CH 2) n-NH-C(＝O)-(CH 2) n-；所述X选自O，NH；所述n，m各自独立的选自0-6。
  2. 根据权利要求1所述的一种式I所示的化合物及其消旋体、立体异构体、互变异构体、同位素标记物、氮氧化物、溶剂化物、多晶型物、代谢产物、酯、前药或其药学上可接受的盐，其特征在于：
    所述L 1、L 2、L 4各自独立的选自-(CH 2) n-C(＝X)-或-C(＝X)-(CH 2) n-，-(CH 2) m-(当m＝0时，代表键)，所述L 3选自-(CH 2) m-(当m＝0时，代表键)；所述L 5选自-NH-C(＝O)-,-C(＝O)-NH-，-(CH 2) m-(当m＝0时，代表键)；所述X选自O，NH；所述n，m各自独立 的选自0，1，2，3，4，5，6；
    优选的，所述L 1选自-(CH 2) 2-，-(CH 2) 3-，-(CH 2) 4-，-(CH 2) 5-，-CH 2-C(＝O)-，-(CH 2) 2-C(＝O)-，-(CH 2) 3-C(＝O)-，-(CH 2) 4-C(＝O)-；L 2选自键，-CH 2-C(＝O)-，-CH 2-，，-C(＝O)-；L 3选自键，-CH 2-；L 4选自键，-CH 2-；L 5选自键，-CH 2-。
  3. 根据权利要求1-2任一项所述的一种式I所示的化合物及其消旋体、立体异构体、互变异构体、同位素标记物、氮氧化物、溶剂化物、多晶型物、代谢产物、酯、前药或其药学上可接受的盐，其特征在于：
    所述“卤素”选自F、Cl、Br、I；所述“脂肪烃基”选自烷基、烯基、炔基；
    优选的，所述R 1、R 5各自独立的选自卤素，C 1-C 6脂肪烃基，例如，选自F、Cl、Br、I，甲基，乙基，正丙基，异丙基，正丁基，异丁基，叔丁基，正戊基，异戊基，新戊基，正己基；
    优选的，所述R 2选自H，C 1-C 6脂肪烃基或C 3-8环烷基-C 1-C 6脂肪烃基，且R 3选自-L 5-Ar或所述R 2选自-L 5-Ar且R 3选自H，C 1-C 6脂肪烃基或C 3-8环烷基-C 1-C 6脂肪烃基；更优选的，所述R 2或R 3之一选自H，甲基，乙基，正丙基，异丙基，正丁基，异丁基，叔丁基，正戊基，异戊基，新戊基，正己基，环丙基甲基，环丁基甲基，环戊基甲基，环己基甲基；
    所述Ar选自任选被一个、两个或更多个R b取代的如下基团：C 6-C 14芳基，5-10元杂环基，5-6元杂芳基，优选的，Ar选自任选被一个、两个或更多个R b取代的苯基，萘基，2,3-二氢苯并呋喃基，苯并呋喃基，苯并吡喃基，3,4-二氢-2H-1-苯并吡喃基(色烷基)，2,3-二氢苯并[b][1,4]二噁烷基，吡啶基，嘧啶基，吲唑基(1H-吲唑基，2H-吲唑基)，吲哚基，异吲哚基，喹啉基，异喹啉基，喹唑啉基，苯并噁唑基；
    优选的，所述R b选自H，卤素，CN，或任选被一个、两个或更多个R c取代的如下基团：C 1-C 6脂肪烃基，C 1-C 6脂肪烃基氧基，C 1-C 6脂肪烃基-SO 2-，C 1-C 6脂肪烃基-NH-，二(C 1-C 6脂肪烃基)N-，C 6-10芳基，C 6-10芳基氧基，C 6-10芳基-SO 2-，C 6-10芳基-NH-，5-6元杂芳基，5-6元杂芳基氧基，5-6元杂芳基-SO 2-，5-6元杂芳基-NH-，5-6元杂环基，5-6元杂环基氧基，5-6元杂环基-SO 2-，5-6元杂环基-NH-，C 3-8环烷基，C 3-8环烷基氧基，C 3-8环烷基-SO 2-，C 3-8环烷基-NH-，C 3-8环烷基-C 1-C 6脂肪烃基氧基，C 3-8环烷基-C 1-C 6脂肪烃基-SO 2-，C 3-8环烷基-C 1-C 6脂肪烃基-NH-；更优选的，所述R b选自H，卤素，CN或任选被一个、两个或更多个R c取代的C 1-C 6烷基，C 1-C 6烷氧基，C 1-C 6烷基-SO 2-，C 1-C 6烷基-NH-，二(C 1-C 6烷基)N-，5-6元杂芳基-SO 2-，5-6元杂芳基-NH-，5-6元杂环基，5-6元杂环基氧基，5-6元杂环基-SO 2-，5-6元杂环基-NH-，C 3-6环烷基，C 3-6环烷基氧基，C 3-6环烷基-SO 2-，C 3-6环烷基-NH-，C 3-6环烷基-C 1-C 6烷氧基，C 3-6环烷基-C 1-C 6烷基-SO 2-，C 3-6环烷基-C 1-C 6烷基-NH-；进一步优选的，所述R b选自H，卤素，CN或任选被一个、两个或更多个R c取代的甲基，乙基，正丙基，异丙基，正丁基，异丁基，叔丁基，正戊基，异戊基，新戊基，正己基，乙烯基，1-丙烯基，2-丙烯基，1-甲基乙烯基，1-丁烯基，1-乙基乙烯基，1-甲基-2-丙烯基，2-丁烯基，3-丁烯基、2-甲基-1-丙烯基，2-甲基-2-丙烯基，1-戊烯基、1-己烯基，乙炔基，1-丙炔基，2-丙炔基，1-丁炔基，1-甲基-2-丙炔基，3-丁炔基，1-戊炔基，1-己炔基，甲氧基，乙氧基，正丙氧基，异丙氧基，异丁氧基，正丁氧基，叔丁氧基，戊氧基，己氧基，N,N-二甲基氨基，N,N-二乙基氨基，甲基-NH-，乙基-NH-，正丙基-NH-，异丙基-NH-，正丁基-NH-，异丁基-NH-，叔丁基-NH-，正戊基-NH-，异戊基-NH-，新戊基-NH-，正己基-NH-， 甲基-SO 2-，乙基-SO 2-，正丙基-SO 2-，异丙基-SO 2-，正丁基-SO 2-，异丁基-SO 2-，叔丁基-SO 2-，正戊基-SO 2-，异戊基-SO 2-，新戊基-SO 2-，正己基-SO 2-，环丙基，环丁基，环戊基，环己基，环丙基氧基，环丁基氧基，环戊基氧基，环己基氧基，环丙基，环丁基，环戊基，环己基，环丙基-SO 2-，环丁基-SO 2-，环戊基-SO 2-，环己基-SO 2-，环丙基-NH-，环丁基-NH-，环戊基-NH-，环己基-NH-，环丙基甲基氧基，环丁基甲基氧基，环戊基甲基氧基，环己基甲基氧基，环丙基甲基-SO 2-，环丁基甲基-SO 2-，环戊基甲基-SO 2-，环己基甲基-SO 2-，环丙基甲基-NH-，环丁基甲基-NH-，环戊基甲基-NH-，环己基甲基-NH-，苯基，苯基-SO 2-，苯基-NH-，萘基，萘基-SO 2-，萘基-NH，氧杂环丁烷，吡喃基，四氢吡喃基，呋喃基，四氢呋喃基，四氢吡咯基，哌啶基，吡啶基，吡嗪基，吡咯基，咪唑基，吡唑基，三氮唑，噁唑基，异噁唑基，吗啉基；例如，R b选自F，Cl，Br，I，CN，甲基，乙基，正丙基，异丙基，正丁基，异丁基，叔丁基，新戊基，甲基-SO 2-，甲氧基，乙氧基，正丙氧基，异丙氧基，异丁氧基，N,N-二甲基氨基，异丙基-NH-，环丙基氧基，环丁基氧基，环戊基氧基，环丙基甲基氧基，环丙基氨基，CF 3，CHF 2，CH 2F，CF 3O，CHF 2O，CH 2FO，苯基，3,5-二甲基吡唑-1-基，咪唑基，吗啉基，1,2,4-三氮唑，氧杂环丁烷，四氢呋喃基，四氢吡喃基，四氢吡咯基；
    优选的，所述R c选自H，＝O，卤素，OH，CN，SH，NH 2，COOH，C 1-C 6烷基，C 1-C 6烷氧基，C 1-C 6烷基-SO 2-，二(C 1-C 6烷基)N-，C 1-C 6烷基-NH-；例如选自H，＝O，F，Cl，Br，I，OH，CN，SH，NH 2，COOH，甲基，乙基，正丙基，异丙基，正丁基，异丁基，叔丁基，正戊基，异戊基，新戊基，正己基，甲氧基，乙氧基，正丙氧基，异丙氧基，异丁氧基，正丁氧基，叔丁氧基，戊氧基，己氧基，N,N-二甲基氨基，N,N-二乙基氨基，甲基-NH-，乙基-NH-，正丙基-NH-，异丙基-NH-，正丁基-NH-，异丁基-NH-，叔丁基-NH-，正戊基-NH-，异戊基-NH-，新戊基-NH-，正己基-NH-，甲基-SO 2-，乙基-SO 2-，正丙基-SO 2-，异丙基-SO 2-，正丁基-SO 2-，异丁基-SO 2-，叔丁基-SO 2-，正戊基-SO 2-，异戊基-SO 2-，新戊基-SO 2-，正己基-SO 2-；
    所述R 4独立的选自H，C 1-C 6脂肪烃基，优选的，选自甲基，乙基，正丙基，异丙基，正丁基，异丁基，叔丁基，正戊基，异戊基，新戊基，正己基。
  4. 根据权利要求1-3任一项所述的一种式I所示的化合物及其消旋体、立体异构体、互变异构体、同位素标记物、氮氧化物、溶剂化物、多晶型物、代谢产物、酯、前药或其药学上可接受的盐，其特征在于，所述Ar选自如下基团：
    Figure PCTCN2021078537-appb-100002
    Figure PCTCN2021078537-appb-100003
    所述结构中，L 5(不为键)，R b，R c选自式I中所述定义。
  5. 根据权利要求1-4任一项所述的一种式I所示的化合物及其消旋体、立体异构体、互变异构体、同位素标记物、氮氧化物、溶剂化物、多晶型物、代谢产物、酯、前药或其药学上可接受的盐，其特征在于，所述R 2或R 3中的一个选自如下基团：
    Figure PCTCN2021078537-appb-100004
    Figure PCTCN2021078537-appb-100005
    Figure PCTCN2021078537-appb-100006
  6. 根据权利要求1-5任一项所述的一种式I所示的化合物及其消旋体、立体异构体、互变异构体、同位素标记物、氮氧化物、溶剂化物、多晶型物、代谢产物、酯、前药或其药学上可接受的盐，其特征在于，所述式I结构可以选自式II：
    Figure PCTCN2021078537-appb-100007
    所述式II结构中，R 1、R 2、R 3、R 4、R 5、L 1、L 2、L 3、L 4以及其他手性中心如上前述式I所定义；
    优选的，所述式II结构进一步如下式IIa-IIt所示：
    Figure PCTCN2021078537-appb-100008
    Figure PCTCN2021078537-appb-100009
    Figure PCTCN2021078537-appb-100010
    Figure PCTCN2021078537-appb-100011
  7. 根据权利要求1-6任一项所述的一种式I所示的化合物及其消旋体、立体异构体、互变异构体、同位素标记物、氮氧化物、溶剂化物、多晶型物、代谢产物、酯、前药或其药学上可接受的盐，其特征在于，所述式I结构可以选自所述式I化合物选自如下式III(式IIIa、式IIIb)、式IV(式IVa、式IVb)：
    Figure PCTCN2021078537-appb-100012
    所述式III(式IIIa、式IIIb)、式IV(式IVa、式IVb)结构中，R 1、R 3、R 4、R 5、L 1、L 3以及其他手性中心如上前述式I所定义；
    优选的，所述式III(式IIIa、式IIIb)、式IV(式IVa、式IVb)结构中，所述L 3选自-(CH 2) m-，且m＝0(即为键)；
    优选的，所述式I结构可以选自如下式V至式XVII：
    Figure PCTCN2021078537-appb-100013
    Figure PCTCN2021078537-appb-100014
    Figure PCTCN2021078537-appb-100015
    所述式V至式XVII结构中，R 1、R 2、R 3、R 4、R 5、n、m如前述式I所定义。
  8. 根据权利要求1-7任一项所述的一种式I所示的化合物及其消旋体、立体异构体、互变异构体、同位素标记物、氮氧化物、溶剂化物、多晶型物、代谢产物、酯、前药或其药学上可 接受的盐，其特征在于，式I选自如下具体化合物：
    Figure PCTCN2021078537-appb-100016
    Figure PCTCN2021078537-appb-100017
    Figure PCTCN2021078537-appb-100018
    Figure PCTCN2021078537-appb-100019
    Figure PCTCN2021078537-appb-100020
    Figure PCTCN2021078537-appb-100021
    Figure PCTCN2021078537-appb-100022
    Figure PCTCN2021078537-appb-100023
    Figure PCTCN2021078537-appb-100024
  9. 根据权利要求1-7任一项所述的一种式I所示的化合物及其消旋体、立体异构体、互变异 构体、同位素标记物、氮氧化物、溶剂化物、多晶型物、代谢产物、酯、前药或其药学上可接受的盐，其特征在于，式I选自如下具体化合物：
    Figure PCTCN2021078537-appb-100025
    Figure PCTCN2021078537-appb-100026
  10. 根据权利要求1-9任一项所述化合物的制备方法，其特征在于，包括如下步骤：在合适的条件下，将含萘啶环结构的原料与含吡咯烷结构的原料在合适的试剂中进行反应，并任选的，在合适的条件下，进行上保护基，脱保护基，还原，胺化，缩合或水解步骤。
  11. 一种药物组合物，其包含权利要求1-9任一项所述的式I化合物及其消旋体、立体异构体、互变异构体、同位素标记物、氮氧化物、溶剂化物、多晶型物、代谢产物、酯、前药或其药学上可接受的盐；优选的，所述药物组合物包含治疗有效量的式I化合物及其消旋体、立体异构体、互变异构体、同位素标记物、氮氧化物、溶剂化物、多晶型物、代谢产物、酯、前药或其药学上可接受的盐和药学上可接受的载体。
  12. 根据权利要求1-9任一项所述的式I化合物及其消旋体、立体异构体、互变异构体、同位素标记物、氮氧化物、溶剂化物、多晶型物、代谢产物、酯、前药或其药学上可接受的盐或所述药物组合物在制备预防，调节或治疗与整合素活性有关的疾病或病症的药物中的用途。
  13. 根据权利要求12所述的用途，其特征在于，所述整合素选自αVβ1，αVβ3，αVβ5，αVβ6，和αVβ8，α5β1，α8β1中的任一种或αVβ1，αVβ3，αVβ5，αVβ6，和αVβ8，α5β1，α8β1中的一个或多个的组合。
  14. 根据权利要求1-9任一项所述式I化合物及其消旋体、立体异构体、互变异构体、同位素标记物、氮氧化物、溶剂化物、多晶型物、代谢产物、酯、前药或其药学上可接受的盐或所述药物组合物在用于抑制细胞中TGF-β活化的药物中的用途。
  15. 根据权利要求1-9任一项所述的式I化合物及其消旋体、立体异构体、互变异构体、同位素标记物、氮氧化物、溶剂化物、多晶型物、代谢产物、酯、前药或其药学上可接受的盐或所述药物组合物在制备用于治疗纤维化性疾病，炎性疾病或细胞增殖性疾病的药物中的用途。
  16. 根据权利要求12或13所述的用途，所述疾病或病症选自移植注射，纤维化病症(例如特发性肺纤维化，间质性肺疾病，肝纤维化，非酒精性脂肪性肝，原发性硬化性胆管炎(PSC)，肾纤维化，皮肤纤维化，心肌纤维化，系统性硬化)，炎症性疾病(例如急性肝炎，慢性肝炎，牛皮癣，肠易激综合征(IBS)，炎症性肠病(IBD)，骨质疏松症以及细胞增殖性疾病(例如 癌症，骨髓瘤，纤维瘤，肝癌，白血病，卡波西氏肉瘤，实体瘤)；
    所述疾病或病症进一步选自特发性肺纤维化(IPF)，间质性肺病，非特异性间质性肺炎(NSIP)，常规间质性肺炎(UIP)，辐射诱发性纤维化，家族性肺纤维化，气道纤维化，慢性阻塞性肺疾病(COPD)，糖尿病性肾病，局灶性节段性肾小球硬化，IgA肾病，药物或移植引起的肾病，自身免疫性肾病，狼疮性肾炎，肝纤维化，肾脏纤维化，慢性肾脏病(CKD)，糖尿病肾病(DKD)，皮肤纤维化，瘢痕，全身性硬化，硬皮病，病毒性纤维化，非酒精性脂肪肝病(NAFLD)，酒精性或非酒精性脂肪性肝炎(NASH)，急性肝炎，慢性肝炎，肝硬化，原发性硬化性胆管炎，药物性肝炎，胆汁性肝硬化，门脉高压，再生衰竭，肝功能不全，肝血流异常，肾病，肺炎，牛皮癣，肠易激综合征罗马(IBS)，炎性肠病(IBD)，胰腺分泌异常，前列腺增生，神经性膀胱疾病，脊髓肿瘤，椎间盘疝，椎管狭窄，心力衰竭，心脏纤维化，血管纤维化，血管周纤维化，足口疾病，癌症，骨髓瘤，纤维瘤，肝癌，白血病，慢性淋巴细胞性白血病，卡波济肉瘤，实体瘤，脑梗死，脑出血，神经性疼痛，周围神经病，年龄相关性黄斑变性(AMD)，青光眼，眼纤维化，角膜瘢痕形成，糖尿病性视网膜病变，增生性玻璃体视网膜病变(PVR)，瘢痕性天疱疮性青光眼滤过手术瘢痕，克罗恩病或系统性红斑狼疮；伤口愈合异常导致瘢痕形成；器官移植后发生纤维化，骨髓纤维化和肌瘤。
PCT/CN2021/078537 2020-03-06 2021-03-01 一种吡咯烷类整合素调节剂及其用途 WO2021175196A1 (zh)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
CN202010153014.2 2020-03-06
CN202010153014 2020-03-06

Publications (1)

Publication Number Publication Date
WO2021175196A1 true WO2021175196A1 (zh) 2021-09-10

Family

ID=77524736

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/CN2021/078537 WO2021175196A1 (zh) 2020-03-06 2021-03-01 一种吡咯烷类整合素调节剂及其用途

Country Status (3)

Country Link
CN (1) CN113354635B (zh)
TW (1) TW202144344A (zh)
WO (1) WO2021175196A1 (zh)

Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2000009503A1 (en) * 1998-08-13 2000-02-24 Merck & Co., Inc. Integrin receptor antagonists
WO2001024797A1 (en) * 1999-10-04 2001-04-12 Merck & Co., Inc. Integrin receptor antagonists
WO2018160522A1 (en) * 2017-02-28 2018-09-07 Lazuli, Inc. Inhibitors of (alpha-v)(beta-6) integrin
CN109996541A (zh) * 2016-09-07 2019-07-09 普利安特治疗公司 N-酰基氨基酸化合物及其使用方法
CN110167934A (zh) * 2016-11-08 2019-08-23 百时美施贵宝公司 作为αV整联蛋白抑制剂的含有环丁烷和含有氮杂环丁烷的单环和螺环化合物

Patent Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2000009503A1 (en) * 1998-08-13 2000-02-24 Merck & Co., Inc. Integrin receptor antagonists
WO2001024797A1 (en) * 1999-10-04 2001-04-12 Merck & Co., Inc. Integrin receptor antagonists
CN109996541A (zh) * 2016-09-07 2019-07-09 普利安特治疗公司 N-酰基氨基酸化合物及其使用方法
CN110167934A (zh) * 2016-11-08 2019-08-23 百时美施贵宝公司 作为αV整联蛋白抑制剂的含有环丁烷和含有氮杂环丁烷的单环和螺环化合物
WO2018160522A1 (en) * 2017-02-28 2018-09-07 Lazuli, Inc. Inhibitors of (alpha-v)(beta-6) integrin

Also Published As

Publication number Publication date
CN113354635B (zh) 2023-04-18
TW202144344A (zh) 2021-12-01
CN113354635A (zh) 2021-09-07

Similar Documents

Publication Publication Date Title
US11639353B2 (en) Cyclobutanes- and azetidine-containing mono and spirocyclic compounds as αV integrin inhibitors
KR102505629B1 (ko) 알파 v 인테그린 억제제로서의 3-치환된 프로피온산
CN106488915B (zh) 作为lsd1抑制剂的环丙胺
TWI768557B (zh) 作為ｈｄａｃ抑制劑之3-烷基-4-醯胺基-雙環[4,5,0]羥肟酸
AU2017359030A1 (en) Pyrrole amides as alpha v integrin inhibitors
WO2020216190A1 (zh) 一种喹唑啉化合物及其在医药上的应用
TW201533042A (zh) αvβ整合素拮抗劑
TW200948817A (en) Aminocyclohexane derivatives
TW200946528A (en) Piperidine derivatives
TW201311674A (zh) 吲唑－及吡咯并吡啶－衍生物及其醫藥用途
CN101602741A (zh) 作为γ-分泌酶抑制剂的取代的N-芳基磺酰基杂环胺
KR20200083529A (ko) 파르네소이드 x 수용체 조정제로서의 알켄 스피로시클릭 화합물
CN113677673B (zh) Trpc6的抑制剂
WO2022194269A1 (zh) 新型egfr降解剂
WO2021175196A1 (zh) 一种吡咯烷类整合素调节剂及其用途
CN113056455A (zh) 异噁唑甲酰胺化合物及其用途
WO2022223039A1 (zh) Sos1降解剂及其制备方法和应用
WO2022022364A1 (zh) 一种吡咯烷类整合素调节剂及其用途
WO2016075457A1 (en) Sulfonyl piperidine derivatives and their use for treating prokineticin mediated gastrointestinal disorders
TWI840348B (zh) G蛋白偶聯受體調節劑
WO2021051827A1 (zh) 一种细胞凋亡蛋白抑制剂及其制备方法和用途

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: 21763979

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: 21763979

Country of ref document: EP

Kind code of ref document: A1