CN113620931B - Androgen receptor inhibitor and application thereof - Google Patents

Androgen receptor inhibitor and application thereof Download PDF

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CN113620931B
CN113620931B CN202111067653.8A CN202111067653A CN113620931B CN 113620931 B CN113620931 B CN 113620931B CN 202111067653 A CN202111067653 A CN 202111067653A CN 113620931 B CN113620931 B CN 113620931B
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CN113620931A (en
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秦冲
陈怡铭
钟腾江
张赛
葛玲
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Ocean University of China
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    • 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
    • C07D401/14—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 three or more hetero rings
    • 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
    • A61P13/08—Drugs for disorders of the urinary system of the prostate
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P15/00—Drugs for genital or sexual disorders; Contraceptives
    • 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
    • A61P17/08—Antiseborrheics
    • 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
    • A61P17/10—Anti-acne agents
    • 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
    • A61P17/14—Drugs for dermatological disorders for baldness or alopecia
    • 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
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents

Abstract

The invention relates to the field of pharmacy, and in particular discloses an Androgen Receptor (AR) regulator combined with an E3 ubiquitin ligase ligand or salt, stereoisomer and a pharmaceutical composition containing the same.

Description

Androgen receptor inhibitor and application thereof
Technical Field
The invention relates to the field of pharmacy, and in particular discloses an Androgen Receptor (AR) regulator combined with an E3 ubiquitin ligase ligand or salt, stereoisomer and a pharmaceutical composition containing the same.
Background
Proteolys-targeting chimera (PROTAC) is a novel strategy for selective knockdown of target proteins by small molecules (Sakamoto KM et al Proc Natl Acad Sci, 2001,98:8554-9.; sakamoto KM et al Methods enzymol 2005; 399:833-847.). PROTAC targets specific proteins and induces their degradation in cells using the ubiquitin-protease system (Zhou P.et al., mol cell.2000;6 (3): 751-756;Neklesa TK et al., pharmacol Ther.2017;174:138-144; lu M.et al., eur J Med chem.2018; 146:251-259;). The normal physiological function of the ubiquitin protease system is responsible for eliminating denatured, mutated or detrimental proteins in the cells. The normal physiological function of the ubiquitin protease system is responsible for eliminating denatured, mutated or detrimental proteins in the cells. Ubiquitin-proteasome system (UPS), also known as ubiquitin-proteasome pathway (UPP), is a common posttranslational regulatory mechanism responsible for protein degradation under normal and pathological conditions (Ardley h. Et al, esays biochem.2005,41,15-30;Komander D.et al, biochem.2012,81,203-229;Grice GL et al, cell rep.2015,12,545-553;Swatek KN et al, cell res.2016,26, 399-422). Ubiquitin is highly conserved in eukaryotic cells, a modified molecule consisting of 76 amino acids, covalently binds and labels a target substrate by a cascade of enzymatic reactions involving the E1, E2 and E3 enzymes. The modified substrate is then recognized by the 26S proteasome complex for ubiquitination-mediated degradation. So far, two E1 enzymes, called UBA1 and UBA6, respectively, have been found. On the other hand, there are approximately 40E 2 enzymes and more than 600E 3 enzymes that provide functional diversity to control the activity of many downstream protein substrates. However, only a limited number of E3 ubiquitin ligases were successfully hijacked for use by the small molecule PROTAC technology: von Hippel-Lindau tumor suppressor protein (VHL), mouse two-minute 2 homolog (MDM 2), apoptosis inhibitor (cIAP) and cereblon (Philipp o.et al., chem. Biol.2017,12, 2570-2578).
Bifunctional compounds consisting of a target protein binding moiety and an E3 ubiquitin ligase binding moiety have been shown to induce proteasome-mediated degradation of selected proteins. Such drug molecules offer the possibility of time-controlled protein expression and are useful in the treatment of related diseases. In recent years, this newly developed method has been widely used in antitumor studies (Lu J.et al, chem biol.2015;22 (6): 755-763;Ottis P.et al, chem biol.2017;12 (4): 892-898; crews CM et al, J Med chem.2018;61 (2): 403-404; neklesa TK et al, pharmacol Ther.2017, 174:138-144K; cermakova et al, molecular, 2018.23 (8); an S. Et al, EBiomedicine,2018.; lebraud H et al, essars biochem.2017;61 (5): 517-527.; sun YH et al, cell Res.2018;28:779-81; toure M.et al, angew Chem Int Ed Engl.2016;55 (6); 1976); 1973). And have been disclosed or discussed in the patent publications, e.g. US20160045607, US20170008904, US20180050021, US20180072711, WO2002020740, WO2014108452, WO2016146985, WO2016149668, WO2016149989, WO2016197032, WO2016197114, WO2017011590, WO2017030814, WO2017079267, WO2017182418, WO2017197036, WO2017197046, WO2017197051, WO2017197056, WO2017201449, WO2017211924, WO2018033556, WO2018071606.
Prostate cancer (PCa) remains the leading cause of cancer-related death in north american men (S.M.Green, mol.Cell.Endocrinol.360 (2012) 3-13). The main driving factor for this disease is the Androgen Receptor (AR), a steroid-induced transcription factor, which regulates many genes that promote tumor growth (N.Lallous, int.J.Mol.Sci.14 (2013), 12496-12519). AR is part of the nuclear receptor family, consisting of an N-terminal domain (NTD), followed by a DNA Binding Domain (DBD) and a C-terminal Ligand Binding Domain (LBD) (G.Jenster, mol.Endocrinol.5 (1991) 1396-1404). The binding signaling nuclear localization of steroids to LBD followed by DBD-mediated AR dimerization and binding of activated receptor dimers to Androgen Response Elements (ARE) on the target gene. Currently used AR antagonists, such as enzalutamide, block AR activation by interfering with the binding of steroids to the androgen binding pocket on LBD (C.E.Bohl, J.Biol.Chem.280 (2005) 37747-37754). Resistance occurs when mutations in the binding pocket render these drugs ineffective, or by expressing constitutively active splice variants (e.g., AR-V7, 6-9) that lack LBD entirely. Despite the improvement in medical treatment over the past 30 years, prostate cancer (PCa) remains an important cause of cancer-related death, next to lung cancer in men in developed countries (Hamdy, F.C.; N.Engl. J.Med.2016,375,1415-1424; litwin, M.S.; J.Am. Med. Assoc.2017,317, 2532-2542). In addition to surgery and radiation therapy, androgen Deprivation Therapy (ADTs) is a first line treatment for patients with high risk local disease of prostate cancer, and second generation antiandrogens such as abiraterone and enzalutamide have been shown to be beneficial in advanced prostate cancer patients (Karantanos, T.; oncogene 2013,32,5501-5511; harris, WP, nat. Clin. Practice. Neurol.2009,6, 76-85.). However, patients who progress to metastatic castration-resistant prostate cancer (mCRPC), a hormone-resistant disease, are subject to high mortality, and there is currently no cure (Narayanan, r.; oncoscience 2017,4,175-177; croder, cm; endocrinology 2018,159,980-993). Androgen Receptor (AR) and its downstream signaling play a critical role in the development and progression of localized and metastatic prostate cancer. (sundecn, h.; previous strategies for successful targeting of AR signaling have focused mainly on blocking androgen synthesis by drugs (such as abiraterone) and on blocking androgen synthesis by AR antagonists (such as enzalutamide and apaluramide (ARN-509)) (Oksala, r.; steroid biochem.mol.biol.2018; watson, pa., nat.Rev.cancer 2015,15,701-711; guo, C.; J.Med.chem.2011,54,7693-7704.; guerrini, A.; J.Med.chem.2014,57,7263-7279.Jung, M.E, J.Med.chem.2010,53,2779-2796, yamamoto, S.; bioorg. Medical chemistry 2012,20,2338-2352.) however, these drugs became ineffective in advanced prostate cancer with AR gene amplification, mutation and alternate splicing (Lottrup, G.; J.Clin.Endocrinol.2013, 98, 2223-9) however, it was apparent that in most CRPC patients, AR protein continued expression, the tumor still relied on the AR signal and thus the target for AR signaling (CRP.21, 35, 35.g.; mu.35, 35).
The compounds of the invention are more potent new AR antagonists than known AR antagonists and antagonize AR by alternative strategies, for example, by degradation of AR. The present application addresses this need.
Disclosure of Invention
It is an object of the present invention to provide a proteolytic targeting chimeric (PROTAC) compound, which functions to recruit a protein of interest to E3 ubiquitin ligase for degradation, by binding an AR antagonist to an E3 ligase ligand, and to provide a method of preparation and use thereof. In particular, the present disclosure provides a PROTAC compound having formula I.
A compound represented by the general formula (I) or a pharmaceutically acceptable salt, stereoisomer thereof,
Figure BDA0003259005860000031
wherein Q is Linker and A is E3 ubiquitin ligase moiety;
r1, R3, R4 are independently selected from H, halogen, -C1-8 alkyl, -C1-8 alkenyl, -C1-8 alkynyl, cycloalkyl, heterocyclyl, aryl, heteroaryl, said-C1-8 alkyl, -C1-8 alkenyl, -C1-8 alkynyl, cycloalkyl, heterocyclyl, aryl, heteroaryl being substituted with a member selected from halogen, hydroxy, -C1-8 alkoxy, cycloalkyl, heterocyclyl, aryl or heteroaryl;
r2 is selected from C, O and S;
R5-R8 are each independently selected from CRd, N, O, S;
q is a bond or a divalent linking group;
A is a targeted E3 ubiquitin ligase ligand.
R1, R3, R4 are further selected from-C1-8 alkyl.
A is selected from
Figure BDA0003259005860000041
/>
Figure BDA0003259005860000042
Wherein R9 is independently selected from hydrogen, halogen, -C1-8 alkyl, -C1-8 alkenyl, -C1-8 alkynyl, cycloalkyl, heterocyclyl, aryl, heteroaryl, -CN, -NO2, -OR5a, -SO2R5a, -COR5a, -CO2R5a, -CONR5aR5b, -C (=NR 5 a) NR5bR5C, -NR5aR5b, -NR5aCOR5b, -NR5aCONR5bR5C, -NR5aCO2R5b, -NR5aSONR5bR5C, -NR5aSO2NR5bR5C, -NR5aSO2R5b, said-C1-8 alkyl, -C1-8 alkenyl, -C1-8 alkynyl, cycloalkyl, heterocyclyl, aryl, heteroaryl each substituted with a member selected from halogen, hydroxy, -C1-8 alkoxy, cycloalkyl, heterocyclyl, aryl, heteroaryl;
r5a, R5b, R5C are each independently selected from hydroxy, -C1-8 alkyl, -C2-8 alkenyl, -C2-8 alkynyl, cycloalkyl, heterocyclyl, aryl, heteroaryl;
n is 0,1,2;
* Is the junction of Q and A.
Wherein A is further selected from
Figure BDA0003259005860000051
Q is selected from- (L1) z- (L2) r- (L3) Q-;
wherein L1 is selected from
Figure BDA0003259005860000052
L2 is selected from
Figure BDA0003259005860000053
/>
Figure BDA0003259005860000054
L3 is selected from
Figure BDA0003259005860000055
Figure BDA0003259005860000056
z is 1;
r is 0 or 1;
q is 0 or 1
* The joint of the left side and the right side of the Q.
Specific Q is
Figure BDA0003259005860000061
A compound of formula (I) or a pharmaceutically acceptable salt, stereoisomer thereof, the compound of formula (I) being selected from table 1:
Table 1:
Figure BDA0003259005860000062
/>
Figure BDA0003259005860000071
/>
Figure BDA0003259005860000081
/>
Figure BDA0003259005860000091
/>
Figure BDA0003259005860000101
/>
Figure BDA0003259005860000111
/>
Figure BDA0003259005860000121
/>
Figure BDA0003259005860000131
/>
Figure BDA0003259005860000141
/>
Figure BDA0003259005860000151
/>
Figure BDA0003259005860000161
in another aspect, the invention also discloses a pharmaceutical composition comprising a compound of formula (I) or a pharmaceutically acceptable salt, stereoisomer, and at least one pharmaceutically acceptable carrier or excipient.
The dosage forms of the medicine can be oral preparations and injection preparations.
In another aspect, the invention discloses methods of inhibiting AR activity comprising administering to a subject a compound disclosed herein, or a pharmaceutically acceptable salt thereof, including a compound of formula (I) or a specific compound exemplified herein.
In another aspect, the invention features a method of treating a disease or condition in a patient, comprising administering to the patient a therapeutically effective amount of a compound of the invention, or a pharmaceutically acceptable salt thereof, as an AR antagonist, wherein the compound "disclosed herein comprises a compound of formula (I) or a specific compound exemplified herein. In some embodiments, the disease or disorder associated with antagonism of AR, such as a disease or disorder is cancer, metastatic bone disease, prostatic hypertrophy, acne vulgaris, seborrhea, hirsutism, androgenic alopecia, precocious puberty, or maleation. Preferably the cancer is selected from prostate cancer, breast cancer, ovarian cancer, bladder cancer, uterine cancer, pancreatic cancer or hepatocellular cancer. Preferably, the disease or disorder is cancer. More preferably, the disease or condition is prostate cancer. Even more preferably, the disease or disorder is castration-resistant prostate cancer (CRPC). Even more preferably, the disease or disorder is metastatic castration-resistant prostate cancer (mCRPC).
In another aspect, the invention discloses the use of a compound of formula 1, or a pharmaceutically acceptable salt, stereoisomer, or pharmaceutical composition containing the compound, in the manufacture of a medicament for use in an anti-androgenic agent.
Unless specifically defined, the compounds and salts provided herein may also contain all isotopes of atoms present in the intermediates or final compounds. Isotopes include those atoms having the same atomic number but different mass numbers.
Those skilled in the art will appreciate that the methods described are not exclusive means by which the compounds provided herein can be synthesized, and that a broad set of synthetic organic reactions can be obtained for potential use in synthesizing the compounds provided herein. Those skilled in the art know how to select and implement the appropriate synthetic route. Suitable synthetic methods for starting materials, intermediates and products can be determined by reference to documents including, for example, the following references: advances in heterocyclic chemistry, volumes 1-107 (Elsevier, 1963-2012); heterocyclic journal of chemistry, volumes 1-49 (heterocyclic journal of chemistry, 1964-2012); carreira et al, eds., synthesis science, volumes 1-48 (2001-2010); katritzky et al (incorporated), comprehensive organofunctional transformations (Pergamon Press, 1996); katritzky et al (ed); integrated organofunctional conversion II (Elsevier, 2 nd edition, 2004); katritzky et al (incorporated), comprehensive heterocyclic chemistry (Pergamon Press, 1984); smith et al, higher organic chemistry: reactions, mechanisms, and structures, 6 th edition (Wiley, 2007); trost et al (eds.), comprehensive organic synthesis (Pergamon Press, 1991).
The preparation of the compounds described herein may involve protection and deprotection of various chemical groups. The need for protection and deprotection and the selection of appropriate protecting groups can be readily determined by one skilled in the art. The chemistry of protecting groups can be found, for example, in t.w. greene and p.g. m.wuts, protecting groups in organic synthesis, 3 rd edition, wiley & Sons, inc.
The reaction may be monitored according to any suitable method known in the art. For example, product formation may be monitored by: spectroscopic means, such as nuclear magnetic resonance spectroscopy (e.g., 1 h or 13 C) Infrared spectroscopy, spectrophotometry (e.g., UV-visible), mass spectrometry, or chromatographic methods such as High Performance Liquid Chromatography (HPLC), liquid chromatography-mass spectrometry (LCMS), or Thin Layer Chromatography (TLC). The compounds may be purified by a wide variety of methods including High Performance Liquid Chromatography (HPLC) and normal phase silica gel chromatography by one skilled in the art.
As used herein, the phrase "optionally substituted" means unsubstituted or substituted. As used herein, the term "substituted" means that a hydrogen atom is removed and replaced by a substituent. It is understood that substitution at a given atom is limited by valence.
Throughout all definitions, the term "C n-m "means a range including endpoints, where n and m are integers and represent carbon numbers. Examples include C 1-14 And C 2-14 Etc.
The term "alkyl" refers to a hydrocarbon group selected from straight and branched chain saturated hydrocarbon groups comprising 1 to 18, such as 1 to 12, further such as 1 to 10, still further such as 1 to 8, or 1 to 6, or 1 to 4 carbon atoms. Examples of alkyl groups containing 1 to 6 carbon atoms (i.e., C1-6 alkyl groups) include, but are not limited to, methyl, ethyl, 1-propyl or n-propyl ("n-Pr"), 2-propyl or isopropyl ("i-Pr"), 1-butyl or n-butyl ("n-Bu"), 2-methyl-1-propyl or isobutyl ("i-Bu"), 1-methylpropyl or sec-butyl ("s-Bu"), 1-dimethylethyl or tert-butyl ("t-Bu"), 1-pentyl, 2-pentyl, 3-pentyl, 2-methyl-2-butyl, 3-methyl-1-butyl, 2-methyl-1-butyl, 1-hexyl, 2-hexyl, 3-hexyl, 2-methyl-2-pentyl, 3-methyl-2-pentyl, 4-methyl-2-pentyl, 3-methyl-3-pentyl, 2, 3-dimethyl-2-butyl, and 3, 3-dimethyl-2-butyl.
As used herein, the term "C", alone or in combination with other terms n-m Alkyl "refers to a saturated hydrocarbon group having n to m carbons that may be straight or branched. Examples of alkyl moieties include, but are not limited to, chemical groups such as: methyl, ethyl, n-propyl, isopropyl, n-butyl, t-butyl, isobutyl, sec-butyl; higher homologs such as 2-methyl-1-butyl, n-pentyl, 3-pentyl, n-hexyl, and 1, 2-trimethylpropyl, and the like. In some embodiments, the alkyl group comprises 1 to 14 carbon atoms, 1 to 13 carbon atoms, 1 to 12 carbon atoms, 1 to 11 carbon atoms, 1 to 10 carbon atoms, 1 to 9 carbon atoms, 1 to 8 carbon atoms, 1 to 7 carbon atoms, 1 to 6 carbon atoms, 1 to 5 carbon atoms, 1 to 4 carbon atoms, 1 to 3 carbon atoms, 1 to 2 carbon atoms.
The term "C", as used herein, alone or in combination with other terms n-m Alkoxy "refers to a group of the formula-O-alkyl, wherein the alkyl has n to m carbons. Examples of alkoxy groups include methoxyEthoxy, propoxy (e.g., n-propoxy and isopropoxy), and tert-butoxy, and the like. In some embodiments, the alkyl group has 1 to 6, 1 to 4, or 1 to 3 carbon atoms.
"halogen" as used herein refers to F, cl, br or I. In some embodiments, the halogen is F, cl or Br. In some embodiments, halogen is F. In some embodiments, the halogen is Cl. In some embodiments, the halogen is Br. In some embodiments, halogen is I.
The term "haloalkyl" as used herein refers to an alkyl group in which one or more hydrogens are replaced with one or more halogen atoms, such as fluorine, chlorine, bromine and iodine. Examples of haloalkyl include, but are not limited to, halogenated C1-8 alkyl, halogenated C1-6 alkyl, or halogenated C1-4 alkyl, such as-CF 3, -CH2Cl, -CH2CF3, -CHCl2, -CF3, and the like.
The term "alkenyl" refers to a hydrocarbon group selected from the group consisting of straight and branched hydrocarbon groups containing at least one c=c double bond and 2 to 18, for example 2 to 8, further for example 2 to 6 carbon atoms. Examples of alkenyl groups such as C2-6 alkenyl include, but are not limited to, vinyl or vinyl, prop-1-enyl, prop-2-enyl, 2-methylpropan-1-enyl, but-2-enyl, but-3-enyl, but-1, 3-dienyl, 2-methylbut-1, 3-diene, 1-hexene, 2-hexene, 3-hexene, 4-hexene, and 1, 3-dioxin.
The term "alkynyl" refers to a hydrocarbon group selected from straight and branched hydrocarbon groups comprising at least one c≡c triple bond and from 2 to 18, for example from 2 to 8, further for example from 2 to 6 carbon atoms. Examples of alkynyl groups such as C2-6 alkynyl include, but are not limited to, ethynyl, 1-propynyl, 2-propynyl (propargyl), 1-butynyl, 2-butynyl, and 3-butynyl.
The term "cycloalkyl" refers to a hydrocarbon group selected from saturated cyclic hydrocarbon groups, including monocyclic and polycyclic (e.g., bicyclic and tricyclic) groups, including fused, bridged or spirocycloalkyl groups.
For example, cycloalkyl groups may contain 3 to 12, such as 3 to 10, further such as 3 to 8, further such as 3 to 6, 3 to 5 or 3 to 4 carbon atoms. Still further, for example, the cycloalkyl group may be selected from the group consisting of 3 to 12, such as 3 to 10Further for example 3 to 8, 3 to 6 carbon atoms. Examples of monocyclic cycloalkyl groups include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, cyclononyl, cyclodecyl, cycloundecyl and cyclododecyl. In particular, saturated monocyclic cycloalkyl groups such as C 3-8 Examples of cycloalkyl groups include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, and cyclooctyl. In a preferred intercalation, the cycloalkyl group is a monocyclic ring containing 3 to 6 carbon atoms (abbreviated to C 3-6 Cycloalkyl) including, but not limited to, cyclopropyl, cyclobutyl, cyclopentyl, and cyclohexyl. Examples of bicyclic cycloalkyl groups include those having 7 to 12 ring atoms arranged as a fused bicyclic ring selected from [4,4 ]]、[4,5]、[5,5]、[5,6]And [6,6 ]]Ring systems, or as compounds selected from bicyclo [2.2.1 ] ]Heptane, bicyclo [2.2.2]Octane and bicyclo [3.2.2]Bridged bicyclic rings of nonane. Other examples of bicyclic cycloalkyl groups include those arranged to be selected from [5,6 ]]And [6,6 ]]Those of the bicyclic ring system.
The term "spirocycloalkyl" refers to a cyclic structure containing carbon atoms and formed by at least two rings sharing one atom. The term "7-to 12-membered spirocycloalkyl" refers to a cyclic structure containing 7 to 12 carbon atoms and formed by at least two rings sharing one atom.
The term "fused cycloalkyl" refers to a bicyclic cycloalkyl as defined herein, which is saturated and formed from two or more rings sharing two adjacent atoms.
The term "bridged cycloalkyl" refers to a cyclic structure containing carbon atoms and formed from two rings sharing two atoms that are not adjacent to each other. The term "7-to 10-membered bridged cycloalkyl" refers to a cyclic structure containing 7 to 12 carbon atoms and formed from two rings sharing two atoms that are not adjacent to each other.
The term "cycloalkenyl" refers to a non-aromatic cyclic alkyl group of 3 to 10 carbon atoms having a single ring or multiple rings and having at least one double bond, preferably 1 to 2 double bonds. In one embodiment, the cycloalkenyl group is cyclopentenyl or cyclohexenyl, 1-cyclopent-1-enyl, 1-cyclopent-2-enyl, 1-cyclopent-3-enyl, 1-cyclohex-1-enyl, 1-cyclohex-2-enyl, 1-cyclohex-3-enyl, cyclohexenyl, preferably cyclohexenyl.
The term "fused cycloalkenyl" refers to a bicyclic cycloalkyl as defined herein, which contains at least one double bond and is formed from two or more rings sharing two adjacent atoms.
The term "cycloalkynyl" refers to a non-aromatic cycloalkyl of 5 to 10 carbon atoms having a single ring or multiple rings and having at least one triple bond.
The term "fused cycloalkynyl" refers to a bicyclic cycloalkyl as defined herein, which contains at least one triple bond and is formed from two or more rings sharing two adjacent atoms.
The term "benzofused cycloalkyl" is a bicyclic fused cycloalkyl in which a 4-to 8-membered monocyclic cycloalkyl ring is fused to a benzene ring. For example, benzofused cycloalkyl is where the wavy line indicates the point of attachment.
The term "benzofused cycloalkenyl" is a bicyclic fused cycloalkenyl in which a 4-to 8-membered monocyclic cycloalkenyl ring is fused to a benzene ring.
The term "benzo-fused cycloalkynyl" is a bicyclic fused cycloalkynyl group in which a 4-to 8-membered monocyclic cycloalkynyl ring is fused to a benzene ring.
Examples of fused cycloalkyl, fused cycloalkenyl, or fused cycloalkynyl include, but are not limited to, bicyclo [1.1.0] butyl, bicyclo [2.1.0] pentyl, bicyclo [3.1.0] hexyl, bicyclo [4.1.0] heptyl, bicyclo [3.3.0] octyl, bicyclo [4.2.0] octyl, decalin, and benzo3-to 8-membered cycloalkyl, benzoC 4-6 cycloalkenyl, 2, 3-dihydro-1H-indenyl, 1, 2,3, 4-tetraaryl, 1, 4-dihydronaphthyl, and the like. Preferred embodiments are 8 to 9 membered fused rings, in the examples described above referring to cyclic structures containing 8 to 9 ring atoms.
As used herein, the term "aryl" refers to an aromatic hydrocarbon group, which may be monocyclic or polycyclic (e.g., having 2 fused rings). The term "C n-m Aryl "refers to aryl groups having n to m ring carbon atoms. Aryl groups include, for example, phenyl, naphthyl, and the like. In some embodiments, aryl groups have 6 to 10 carbon atoms. In some embodiments, aryl is substituted or unsubstituted phenyl.
As used hereinAs used herein, "cycloalkyl" refers to non-aromatic cyclic hydrocarbons including cyclized alkyl and/or alkenyl groups. Cycloalkyl groups may include monocyclic or polycyclic (e.g., having 2 fused rings) groups. Cycloalkyl groups can have 3, 4, 5, 6 ring carbons (i.e., C 3-6 Cycloalkyl). The ring-forming carbon atom of the cycloalkyl group may be optionally substituted with an oxy group (oxo) or a thio group (sulfodo) (e.g., C (=o) or C (=s)). Examples of cycloalkyl groups include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclopentenyl, cyclohexenyl, or cyclohexadienyl, and the like. In some embodiments, the cycloalkyl is selected from the group consisting of cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl. In some embodiments, cycloalkyl groups have 3 to 6 ring-forming carbon atoms (i.e., C 3-6 Cycloalkyl).
As used herein, "heterocycloalkyl" refers to a non-aromatic, monocyclic or polycyclic heterocycle having one or more ring-forming heteroatoms selected from O, N or S. Included in heterocycloalkyl groups are monocyclic 4-, 5-and 6-membered heterocycloalkyl groups. Examples of heterocycloalkyl groups include pyrrolidin-2-one, 1, 3-isoxazolidin-2-one, pyranyl, tetrahydropyranyl, oxetanyl, azetidinyl, morpholino, thiomorpholino, piperazinyl, tetrahydrofuranyl, tetrahydrothienyl, piperidinyl, pyrrolidinyl, isoxazolidinyl, isothiazolidinyl, pyrazolidinyl, oxazolidinyl, thiazolidinyl, imidazolidinyl, azaalkyl, and the like. The ring-forming carbon atoms and heteroatoms of the heterocycloalkyl group may be optionally substituted by oxo (=o). Heterocycloalkyl groups can be attached through a ring-forming carbon atom or a ring-forming heteroatom. In some embodiments, the heterocycloalkyl group contains 0 to 3 double bonds.
The compounds disclosed herein may contain asymmetric centers and thus may exist as enantiomers. "enantiomer" refers to two stereoisomers of a compound that are non-superimposable mirror images of each other. When the compounds disclosed herein have two or more asymmetric centers, they may also exist as diastereomers. Enantiomers and diastereomers belong to a broader class of stereoisomers. All such possible stereoisomers are intended to be included, such as substantially pure resolved enantiomers, racemic mixtures thereof and mixtures of diastereomers. It is intended to include all stereoisomers of the compounds disclosed herein and/or pharmaceutically acceptable salts thereof. Unless specifically mentioned otherwise, reference to one isomer applies to any possible isomer. When the isomer composition is not specified, all possible isomers are included.
As used herein, the term "substantially pure" means that the target stereoisomer contains no more than 35%, such as no more than 30%, further such as no more than 25%, even further such as no more than 20%, by weight of any other stereoisomer. In some embodiments, the term "substantially pure" means that the target stereoisomer comprises no more than 10 wt%, such as no more than 5 wt%, such as no more than 1 wt% of any other stereoisomer.
When the compounds disclosed herein contain olefinic double bonds, such double bonds are intended to include E and Z geometric isomers unless specified otherwise.
When the compounds disclosed herein comprise a disubstituted ring system, the substituents found on such ring systems may take on both cis and trans structures. Cis-formation means that both substituents are located on the upper side of the 2 substituent positions on the carbon, while trans-form means that they are located on opposite sides. For example, the disubstituted ring system may be a cyclohexyl or cyclobutyl ring.
It may be advantageous to separate the reaction products from each other and/or from the starting materials. The desired product of each step or series of steps is isolated and/or purified (hereinafter isolated) to the desired degree of homogeneity by techniques commonly used in the art. Typically, such separations involve multiphase extraction, crystallization from a solvent or solvent mixture, distillation, sublimation, or chromatography. Chromatography may involve a variety of methods, such as: reversed and normal phases; size exclusion; ion exchange; high, medium and low pressure liquid chromatography methods and apparatus; small-scale analysis; simulated moving bed ("SMB") and preparative thin-layer or thick-layer chromatography, and small-scale thin-layer and flash chromatography techniques. Those skilled in the art will apply techniques most likely to achieve the desired separation.
"diastereoisomers" refers to stereoisomers of a compound having two or more chiral centers that are not mirror images of each other. Mixtures of diastereomers can be separated into their individual diastereomers based on their physical or chemical differences by methods well known to those skilled in the art, such as by chromatography and/or fractional crystallization. Enantiomers can be separated into the corresponding pure enantiomers by reaction with a suitable optically active compound (e.g., a chiral auxiliary such as a chiral alcohol or Mosher acid chloride) to convert the enantiomeric mixture into a diastereomeric mixture, separate the diastereomers, and convert (e.g., hydrolyze) the individual diastereomers. Enantiomers and diastereomers may also be separated by use of a chiral HPLC column.
Single stereoisomers, such as substantially pure enantiomers, may be obtained by resolution of the racemic mixture using methods such as the formation of diastereomers using optical resolution agents (Eliel, e. And Wilen, s.stephaochemistry of Organic compounds, new york: john Wiley & Sons, inc., 1994; lochmiller, CH et al "chromatographic resolution of enantiomers: selectivity review". J.chromatogrj., 113 (3) (1975): pages 283-302) the racemic mixture of the chiral compounds of the invention may be separated and isolated by any suitable method, including: (1) forming ions, diastereomeric salts with chiral compounds and separating by fractional crystallization or other methods, (2) forming diastereomeric compound derivatizing reagents with chiral compounds, separation of diastereomers and conversion to pure stereoisomers, and (3) directly separating the substantially pure or enriched stereoisomers under chiral conditions. See: wei Na, euro W. Stereochemistry of drug: analytical methods and pharmacology. New York: marcel Dekker, inc. All compounds and pharmaceutically acceptable salts thereof may be found with other substances such as water and solvents (e.g., hydrates and solvates), or may be isolated.
In some embodiments, the preparation of the compounds may involve the addition of an acid or base, thereby affecting, for example, the catalysis of the desired reaction or the formation of salt forms, such as acid addition salts.
Examples of the acid may be an inorganic acid or an organic acid and include, but are not limited to, strong acids and weak acids. Some examples of acids include hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, p-toluenesulfonic acid, 4-nitrobenzoic acid, methanesulfonic acid, benzenesulfonic acid, trifluoroacetic acid and nitric acid. Some weak acids include, but are not limited to, acetic acid, propionic acid, butyric acid, benzoic acid, pyroglutamic acid, tartaric acid, valeric acid, caproic acid, enanthic acid, caprylic acid, pelargonic acid, and capric acid (which may be added as may be continued depending on the development).
Examples of the base include lithium hydroxide, sodium hydroxide, potassium hydroxide, lithium carbonate, sodium carbonate, potassium carbonate, and sodium bicarbonate. Some examples of strong bases include, but are not limited to, hydroxides, alkoxides, metal amides, metal hydrides, metal dialkylamides, and arylamines, wherein alkoxides include lithium, sodium, and potassium salts of methyl, ethyl, and t-butyl oxides; metal amides include sodium amide, potassium amide and lithium amide; metal hydrides include sodium hydride, potassium hydride and lithium hydride; and metal dialkylamides include lithium, sodium and potassium salts of methyl, ethyl, n-propyl, isopropyl, n-butyl, t-butyl, trimethylsilyl and cyclohexyl substituted amides.
The phrase "pharmaceutically acceptable" is used herein to refer to those compounds, materials, compositions, and/or dosage forms which are, within the scope of sound medical judgment, suitable for use in contact with the tissues of human beings and animals without excessive toxicity, irritation, allergic response, or other problem or complication, commensurate with a reasonable benefit/risk ratio.
The present application also includes pharmaceutically acceptable salts of the compounds described herein. As used herein, "pharmaceutically acceptable salts" refers to derivatives of the disclosed compounds wherein the parent compound is modified by converting an existing acid or base moiety to its salt form. Examples of pharmaceutically acceptable salts include, but are not limited to, inorganic or organic acid salts of basic residues such as amines; and alkali metal salts or organic salts of acidic residues such as carboxylic acids. Pharmaceutically acceptable salts of the present application include, for example, conventional non-toxic salts of the parent compound formed from non-toxic inorganic or organic acids, including primarily inorganic acid salts such as sulfuric acid, nitric acid, hydrobromic acid, phosphoric acid, hydrochloric acid, boric acid, sulfamic acid and the like; or organic acids such as acetic acid, propionic acid, butyric acid, camphoric acid, caproic acid, caprylic acid, carbonic acid, cinnamic acid, glycolic acid, trifluoroacetic acid, adipic acid, alginic acid, 2-hydroxypropionic acid, 2-oxopropionic acid, stearic acid, lactic acid, citric acid, oxalic acid, malonic acid, succinic acid, pyroglutamic acid, ascorbic acid, aspartic acid, phenylacetic acid, glutamic acid, benzoic acid, salicylic acid, hydroxymaleic acid, palmitic acid, cinnamic acid, isobutyric acid, lauric acid, mandelic acid, maleic acid, fumaric acid, malic acid, tartaric acid, sulfanilic acid, 2-acetoxy-benzoic acid, 2-hydroxy-1, 2, 3-propanetrioic acid, suberic acid, gluconic acid, glucuronic acid, glutamic acid, glutaric acid, formic acid, fumaric acid, mucic acid, gentisic acid, pyruvic acid, salicylic acid, methanesulfonic acid, ethanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid, cyclohexylsulfinic acid, hydroxyethanesulfonic acid, ethanedisulfonic acid, 4- (methoxycarbonyl) butyric acid, dichloroacetic acid, 1, 2-ethanedisulfonic acid, camphor-10-sulfonic acid, 2, 4-dihydroxybenzoic acid, 2-hydroxybenzoic acid, 2-hydroxy-alpha-valeric acid, 2-hydroxy-naphtalene, and the like. Pharmaceutically acceptable salts of the present application can be synthesized from the parent compound containing a basic or acidic moiety by conventional chemical methods. In general, such salts can be prepared by reacting the free acid or base forms of these compounds with stoichiometric amounts of the appropriate base or acid in water or in an organic solvent or in a mixture of both; in general, non-aqueous media such as ether, ethyl acetate, alcohols (e.g., methanol, ethanol, isopropanol, or butanol) or acetonitrile (MeCN) are preferred.
In some embodiments, the compounds and salts provided herein are substantially isolated. By "substantially isolated" is meant that the compound is at least partially or substantially separated from the environment in which it is formed or detected. Partial isolation may include, for example, a composition enriched in the compounds provided herein. Substantial isolation may include compositions comprising at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 90%, at least about 95%, at least about 97%, or at least about 99% by weight of a compound provided herein or a salt thereof. Methods for separating compounds and their salts are conventional in the art.
Pharmaceutical compositions comprising the compounds disclosed herein may be administered to a subject in need thereof by oral, inhalation, rectal, parenteral or topical routes. For oral administration, the pharmaceutical composition may be a conventional solid preparation such as a tablet, powder, granule, capsule, etc., a liquid preparation such as an aqueous or oily suspension or other liquid preparation such as syrup, solution, suspension, etc.; for parenteral administration, the pharmaceutical composition may be: tablets, pills, powders, troches, sachets, cachets, elixirs, suspensions, emulsions, solutions, syrups, aerosols (as a solid or in a liquid medium), ointments, soft and hard gelatin capsules, suppositories, sterile injectable solutions and sterile packaged powders. Preferably, the formulation of the pharmaceutical composition is selected from the group consisting of tablets, coated tablets, capsules, suppositories, nasal sprays or injections, more preferably tablets or capsules. The pharmaceutical composition may be administered in a single unit with an accurate dosage. In preparing the compositions provided herein, the active ingredient is typically admixed with an excipient, diluted by an excipient, or enclosed within such a carrier in the form of, for example, a capsule, sachet, paper, or other container. When the excipient serves as a diluent, it may be a solid, semi-solid, or liquid material, which acts as a vehicle, carrier, or medium for the active ingredient. In addition, the pharmaceutical composition may further comprise additional active ingredients.
All formulations of the pharmaceutical compositions disclosed herein can be produced by methods conventional in the pharmaceutical arts. For example, the active ingredient may be mixed with one or more excipients and then formulated into the desired formulation. By "pharmaceutically acceptable excipient" is meant a conventional pharmaceutical carrier suitable for the desired pharmaceutical formulation, such as: diluents, carriers such as water, various organic solvents, etc., fillers such as starch, sucrose, etc., binders such as cellulose derivatives, alginates, gelatin, and polyvinylpyrrolidone (PVP); wetting agents such as glycerin; disintegrating agents such as agar, calcium carbonate, and sodium bicarbonate; absorption accelerators such as quaternary ammonium compounds; surfactants such as cetyl alcohol; absorption carriers such as kaolin and bentonite; talc powder, calcium stearate, magnesium stearate, polyethylene glycol and other lubricants. In addition, the pharmaceutical composition also comprises dispersing agents, stabilizing agents, thickening agents, complexing agents, buffering agents, penetrating agents and other pharmaceutically acceptable excipients. Enhancers, polymers, fragrances, sweeteners, dyes, and the like.
The term "disease" refers to any disease, disorder, condition, symptom, or indication, and is interchangeable with the term "disorder" or "condition.
The disease or disorder is associated with antagonism of AR, such as cancer, metastatic bone disease, prostatic hypertrophy, acne vulgaris, seborrhea, hirsutism, androgenic alopecia, precocious puberty or maleation. Preferably the cancer is selected from prostate cancer, breast cancer, ovarian cancer, bladder cancer, uterine cancer, pancreatic cancer or hepatocellular cancer. Preferably, the disease or disorder is cancer. More preferably, the disease or condition is prostate cancer. Even more preferably, the disease or disorder is castration-resistant prostate cancer (CRPC). Even more preferably, the disease or disorder is metastatic castration-resistant prostate cancer (mCRPC).
The compounds of the present invention may be used alone or in combination with other therapeutic agents for the treatment of the diseases or conditions of the present invention. The compounds of the invention are combined with other antitumor agents. Such anti-neoplastic agents include, but are not limited to: cyclophosphamide, nitrogen mustard, marflange, bufanin, carmustine, metal platins such as carboplatin, cisplatin, oxaliplatin, camptothecin, irinotecan, daunorubicin, doxorubicin, bleomycin, plicamycin, paclitaxel, vinorelbine, docetaxel, doxorubicin, fluorouracil, methotrexate, cytarabine, gemcitabine, EGFR inhibitors, VEGFR inhibitors, ALK inhibitors, BTK inhibitors, mTOR inhibitors, HDAC inhibitors.
The active ingredient may be effective over a wide dosage range and is generally administered in a pharmaceutically effective amount. However, it will be appreciated that the amount of the compound actually administered will generally be determined by a physician, in light of the relevant circumstances, including the condition to be treated, the chosen route of administration, the actual compound administered, the age, weight and response of the individual subject, the severity of the symptoms of the subject, and the like.
The invention has the beneficial effects of providing a proteolytic targeting chimeric (PROTAC) compound by combining an AR antagonist with an E3 ligase ligand, thereby providing a novel chemical entity for the treatment of various cancers related to AR, and thus converting the current cancer treatment method.
Drawings
FIG. 1 shows the effect of a compound on the reduction of Androgen Receptor (AR) expression level in LNCaP cells.
FIG. 2 shows the effect of a compound on the reduction of Androgen Receptor (AR) expression in VCaP cells.
FIG. 3 shows the effect of a compound on the reduction of Androgen Receptor (AR) expression level in 22RV1 cells.
FIG. 4, proliferation assay of LNCaP cells.
Detailed Description
The present invention will be described in more detail by way of specific examples. The following examples are provided for illustrative purposes and are not intended to limit the invention in any way. Those skilled in the art will readily recognize a wide variety of non-critical parameters that may be altered or modified to achieve substantially the same result.
The examples provided below further illustrate and exemplify the compounds of the present invention and methods of making the same. It should be understood that the scope of the following examples and preparations is not intended to limit the scope of the present invention in any way.
The following synthetic routes describe the preparation of the compounds of the present invention, all starting materials being prepared by the methods described in these routes, by methods well known to those of ordinary skill in the art of organic chemistry, or are commercially available. All of the final compounds of the present invention are prepared by the methods described in these schemes or by methods analogous thereto, which are well known to those of ordinary skill in the art of organic chemistry. All variable factors applied in these routes are as defined below or as defined in the claims.
The present invention will be described in detail by way of specific examples, but the purpose and purpose of these exemplary embodiments are merely to illustrate the present invention, and are not intended to limit the actual scope of the present invention in any way.
Example 1:
Figure BDA0003259005860000251
Figure BDA0003259005860000261
raw material 1 (1.5 g,1.0 eq.) and raw material 2 (1.8 g,1.2 eq.) were dissolved in 40mL methanol, 1 drop of concentrated hydrochloric acid was added, the temperature was raised to 90 ℃ and reacted for 6h, tlc detected complete reaction of raw materials, the reaction solution was concentrated, and purified by column chromatography (V Petroleum ether /V Acetic acid ethyl ester =3/1) to give 2.3g of a white solid in 85% yield. UPLC-MS calculated for C 16 H 24 N 4 O 4 [M+H] + :337.18,found:337.19.UPLC-retention time:6.9min. 1 H NMR(400MHz,Chloroform-d)δ8.84(s,2H),4.34(q,J＝7.1Hz,2H),4.11(s,2H),3.92(s,4H),2.71(t,J＝13.1Hz,2H),2.47(s,3H),2.22(s,1H),1.71(dd,J＝50.0,18.9Hz,4H),1.46(s,9H),1.41–1.26(m,4H),1.11(d,J＝11.7Hz,2H).
Figure BDA0003259005860000262
Intermediate 3 (2.3 g,1.0 eq.) was dissolved in 60mL dichloromethane, 15mL 4m dioxane solution of hydrochloric acid was added, stirred overnight at room temperature, the starting material was detected by TLC, the reaction was complete, filtered, the solid was dissolved in 30mL water, pH was adjusted to 8-9 with aqueous ammonia, extracted with dichloromethane, dried over anhydrous magnesium sulfate, filtered and concentrated to a white solid 1.5g, yield 95%. UPLC-MS calculated for C 11 H 16 N 4 O 2 [M+H] + :237.13,found:237.14.UPLC-retention time:4.2min.
Figure BDA0003259005860000263
Intermediate 4 (1.7 g,1.0 eq.) and intermediate 5 (1.9 g,1.2 eq.) were dissolved in 50mL 1, 2-dichloroethane, 2 drops of acetic acid were added, stirred at room temperature for 0.5h, sodium triacetylborohydride (3.0 g,2.0 eq.) was added, reacted overnight at room temperature, TLC detected complete starting material reaction, quenched with 20mL methanol, the reaction mixture concentrated, and purified by column chromatography (V Dichloromethane (dichloromethane) /V Methanol =20/1) to give 2.7g of a white solid in 86% yield. UPLC-MS calculated for C 22 H 35 N 5 O 4 [M+H] + :434.27,found:434.28.UPLC-retention time:6.1min. 1 H NMR(400MHz,CDCl 3 )δ8.84(s,2H),4.34(q,J＝7.1Hz,2H),4.11(s,2H),3.92(s,4H),2.71(t,J＝13.1Hz,2H),2.47(s,3H),2.22(s,1H),1.71(dd,J＝50.0,18.9Hz,4H),1.46(s,9H),1.41–1.33(m,3H),1.26(s,1H),1.11(d,J＝11.7Hz,2H).
Figure BDA0003259005860000271
Intermediate 6 (300 mg,1.0 eq.) was dissolved in 20mL of a mixed solution of tetrahydrofuran and water (V Tetrahydrofuran (THF) /V Water and its preparation method =3/1), heating to 35 ℃ and reacting overnight, TLC detecting that the raw materials are completely reacted, adjusting the pH of the reaction liquid to about 5.0 by using 1M HCl, extracting by using dichloromethane, and concentrating the organic phase by drying to obtain 270mg of white solid with the yield of 96%. UPLC-MS calculated for C 20 H 31 N 5 O 4 [M+H] + :406.24,found:406.25.UPLC-retention time:3.7min.
Figure BDA0003259005860000272
Intermediate 7 (170 mg,1.0 eq.) raw material 8 (102 mg,1.2 eq.) HATU (316 mg,1.2 eq.) was dissolved in 10mL anhydrous N, N-dimethylformamide, argonGas protection, addition of N, N-diisopropylethylamine (360. Mu.L, 3.0 eq.) and reaction at room temperature overnight, TLC detection, complete reaction of the starting material, addition of 20mL of water, extraction with ethyl acetate, drying the organic phase with anhydrous sodium sulfate, concentration, column chromatography purification (V Petroleum ether /V Acetic acid ethyl ester =3/1) to give 190mg of a white solid, yield 89%. UPLC-MS calculated for C 27 H 38 N 6 O 4 [M+H] + :511.30,found:511.31.UPLC-retention time:5.5min.
Figure BDA0003259005860000273
Intermediate 9 (190 mg,1.0 eq.) was dissolved in 8mL anhydrous N, N-dimethylformamide, 60% nah (23 mg,1.5 eq.) was added under ice-bath conditions, after stirring for 0.5h, starting material 10 (70 mg,1.2 eq.) was added, the reaction was allowed to proceed overnight at room temperature, TLC detection, starting material reaction was complete, 20mL water was added to the reaction solution, extraction was performed with ethyl acetate, the organic phase was dried over anhydrous sodium sulfate, concentrated, and purified by column chromatography (V Dichloromethane (dichloromethane) /V Methanol =40/1-20/1) to yield 180mg of white solid in 75%. UPLC-MS calculated for C 34 H 40 ClN 7 O 4 [M+H] + :646.28,found:646.00.UPLC-retention time:5.2min. 1 H NMR(400MHz,CDCl 3 )δ8.80(s,2H),8.23(d,J＝22.4Hz,1H),7.64–7.44(m,3H),6.93(d,J＝13.4Hz,2H),6.74(dd,J＝58.9,8.5Hz,1H),4.07(d,J＝12.7Hz,2H),3.90(s,4H),2.70(d,J＝13.2Hz,2H),2.45(s,4H),2.20(d,J＝6.9Hz,2H),2.11(s,3H),1.79–1.57(m,3H),1.43(s,9H),,1.07(q,J＝12.1Hz,2H).
Figure BDA0003259005860000281
Dissolving intermediate 11 (180 mg,1.0 eq.) in 5mL of dichloromethane, adding 1mL of dioxane hydrochloride, stirring overnight at room temperature, detecting complete reaction of raw materials by TLC, filtering, dissolving solid in 10mL of water, adjusting pH to 8-9 with ammonia water, extracting with dichloromethane, adding anhydrous magnesium sulfate, drying, filtering, concentrating to obtain white solid Body 146mg, yield 96%. UPLC-MS calculated for C 29 H 32 ClN 7 O 2 [M+H] + :546.23,found:545.92.UPLC-retention time:4.4min.
Figure BDA0003259005860000282
Intermediate 12 (66 mg,1.0 eq.) and intermediate 15 (40 mg,1.2 eq.) were dissolved in 5mL anhydrous N, N-dimethylformamide, protected with argon, N-diisopropylethylamine (40 μl,2.0 eq.) was added, the reaction was allowed to proceed to 90 ℃ for 8h, tlc detected complete reaction of starting materials, the reaction concentrated, and purified by column chromatography (V Dichloromethane (dichloromethane) /V Methanol =80/1-30/1), 63mg of fluorescent green solid was obtained in 65% yield. UPLC-MS calculated for C 42 H 40 ClN 9 O 6 [M+H] + :802.28found:802.05.
UPLC-retention time:5.9min. 1 H NMR(400MHz,DMSO-d 6 )δ11.06(s,1H),10.23(s,1H),8.95(s,2H),7.91(d,J＝8.6Hz,1H),7.73(d,J＝1.8Hz,2H),7.65(d,J＝8.8Hz,1H),7.34(s,1H),7.16(s,1H),7.10(d,J＝8.6Hz,1H),6.91(dd,J＝8.4,2.2Hz,1H),5.09(d,J＝11.7Hz,1H),4.09(d,J＝12.8Hz,2H),3.04(m,8H),2.97(m,1H),2.58-2.47(m,5H),1.97(m,2H),1.82(d,J＝12.8Hz,2H),1.26(m,2H),1.20(m,4H).
Figure BDA0003259005860000283
Raw material 13 (3.0 g,1.0 eq.) was dissolved in 60mL acetic acid, raw material 14 (3.27 g,1.1 eq.) and potassium acetate (5.5 g,3.1 eq.) were added sequentially, heated to 90 ℃ and stirred for 14h, TLC detected complete consumption of raw material, the reaction solution cooled to room temperature, poured into water, extracted with dichloromethane, dried and concentrated, purified by column chromatography (V Dichloromethane (dichloromethane) /V Methanol =40/1-20/1) to give 4.5g of a white solid in 95% yield. UPLC-MS calculated for C 13 H 9 FN 2 O 4 [M+H] + :277.05found:277.15.UPLC-retention time:5.0min. 1 H NMR(400MHz,DMSO-d 6 )δ11.12(s,1H),7.98(dd,J＝8.3,4.5Hz,1H),7.82(dd,J＝7.0,1.7Hz,1H),7.69(td,J＝9.3,8.6,2.2Hz,1H),5.17–5.08(m,1H),2.94–2.78(m,1H),2.67–2.46(m,2H),2.09–1.96(m,1H).
Example 2:
Figure BDA0003259005860000291
intermediate 12 (66 mg,1.0 eq.) and intermediate 16 (43 mg,1.2 eq.) were dissolved in 5mL anhydrous N, N-dimethylformamide, protected with argon, N-diisopropylethylamine (44 μl,2.0 eq.) was added, the reaction was allowed to proceed to 90 ℃ for 5h, tlc detected complete reaction of starting materials, the reaction concentrated, and purified by column chromatography (V Dichloromethane (dichloromethane) /V Methanol =80/1-30/1), 79mg of fluorescent green solid was obtained in 80% yield. UPLC-MS calculated for C 42 H 39 ClFN 9 O 6 [M+H] + :820.27found:819.93.
UPLC-retention time:5.8min. 1 H NMR(400MHz,DMSO-d 6 )δ11.09(s,1H),10.23(s,1H),8.95(s,2H),7.91(dd,J＝8.7,1.3Hz,2H),7.72(d,J＝11.3Hz,2H),7.46(s,1H),7.16(s,1H),6.91(d,J＝11.1Hz,1H),4.93(dd,J＝121.4,13.3Hz,2H),3.63(d,J＝12.0Hz,4H),3.10(m,4H),2.96–2.83(m,2H),2.56(d,J＝18.2Hz,1H),2.08(s,3H),2.11-1.86(m,8H),1.38(d,J＝13.0Hz,2H),1.20(m,1H).
Figure BDA0003259005860000292
Raw material 15 (3.3 g,1.0 eq.) was dissolved in 60mL acetic acid, raw material 14 (3.27 g,1.1 eq.) and potassium acetate (5.5 g,3.1 eq.) were added sequentially, heated to 90 ℃ and stirred for 14h, TLC detected complete consumption of raw material, the reaction solution cooled to room temperature, poured into water, extracted with dichloromethane, dried and concentrated, purified by column chromatography (V Dichloromethane (dichloromethane) /V Methanol =40/1-20/1), 5.0g of white solid was obtained in 95% yield. UPLC-MS calculated for C 13 H 8 F 2 N 2 O 4 [M+H] + :295.05found:295.06.UPLC-retention time:5.0min. 1 H NMR(400MHz,DMSO-d 6 )δ11.13(s,1H),8.13(t,J＝7.8Hz,2H),5.19–5.09(m,1H),2.85(t,J＝13.7Hz,1H),2.62–2.47(m,2H),2.07–1.99(m,1H).
Example 3:
Figure BDA0003259005860000301
raw material 18 (5.0 g,1.0 eq.) and intermediate 2 (3.8 g,1.0 eq.) were dissolved in 15mL 1, 2-dichloroethane, 2 drops of acetic acid were added, stirred at room temperature for 0.5h, then sodium triacetylborohydride (8.6 g,2.0 eq.) was added, reacted overnight at room temperature, TLC detected complete raw material reaction, quenched with 10mL methanol, the reaction mixture was concentrated, and purified by column chromatography (V Dichloromethane (dichloromethane) /V Methanol =30/1) to give 7.5g of a white solid, yield 89%. UPLC-MS calculated for C 23 H 35 N 3 O 4 [M+H] + :418.27,found:418.15.UPLC-retention time:2.5min. 1 H NMR(400MHz,DMSO-d 6 )δ7.31(s,5H),6.02(s,2H),3.96(d,J＝12.0Hz,2H),2.22(t,J＝4.0Hz,4H),2.07(t,J＝4.0Hz,2H),1.66(s,1H),1.63(s,2H),1.36(s,9H),0.97-0.87(m,2H).
Figure BDA0003259005860000311
Intermediate 19 (7.5 g) was dissolved in 150mL methanol, 10% pd (0.75 g) was added, hydrogen was displaced 3 times, the temperature was raised to 50 ℃, the reaction was carried out for 4h, tlc monitored the starting material was complete, 10% pd was removed by suction filtration, and the filtrate was concentrated to give 4.9g of a white solid in 100% yield. UPLC-MS calculated for C 15 H 29 N 3 O 2 [M+H] + :284.22,found:284.02.UPLC-retention time:1.8min.
Figure BDA0003259005860000312
Intermediate 20 (1.3 g,1.2 eq.) and raw material 21 (0.7 g,1.0 eq.) were dissolved in 40mL of isopropanol, 1 drop of concentrated hydrochloric acid was added, and the temperature was raised toThe reaction was carried out at 90℃for 6h, the TLC detection starting material was complete, the reaction solution was concentrated, and purified by column chromatography (V Petroleum ether /V Acetic acid ethyl ester =3/1) to give 1.5g of a white solid, yield 89%. UPLC-MS calculated for C 22 H 35 N 5 O 4 [M+H] + :434.27,found:434.10.UPLC-retention time:6.2min. 1 H NMR(400MHz,DMSO-d 6 )δ7.88(s,2H),4.50(q,J＝8.0Hz,2H),4.23(d,J＝8.0Hz,1H),3.63(d,J＝8.0Hz,1H),2.82(t,J＝8.0Hz,3H),2.79(t,J＝8.0Hz,1H),2.59(d,J＝8.0Hz,1H),2.29-2.26(m,4H),1.98-1.1.91(m,2H),1.39(dd,J＝8.0Hz,4.0,2H),1.37(s,2H),1.36(s,9H),1.19(m,2H),0.97-0.87(m,2H).
Figure BDA0003259005860000313
Intermediate 22 (500 mg,1.0 eq.) was dissolved in 20mL of a mixed solution of tetrahydrofuran and water (V Tetrahydrofuran (THF) /V Water and its preparation method =3/1), heating to 35 ℃ and reacting overnight, TLC detecting that the raw materials are completely reacted, adjusting the pH of the reaction liquid to about 5.0 by using 1M HCl, extracting by using dichloromethane, and concentrating the organic phase by drying to obtain 420mg of white solid with the yield of 90%. UPLC-MS calculated for C 20 H 31 N 5 O 4 [M+H] + :406.24,found:406.10.UPLC-retention time:1.8min.
Figure BDA0003259005860000314
Intermediate 23 (270 mg,1.0 eq.) and HATU (304 mg,1.2 eq.) were dissolved in 10mL of anhydrous N, N-dimethylformamide under argon, N-diisopropylethylamine (350 μl,2.0 eq.) was added, stirred at room temperature for 0.5h, intermediate 26 (216 mg,1.1 eq.) was added, reacted at room temperature for 6h, tlc was detected, the starting material was complete, 20mL of water was added, extracted with ethyl acetate, the organic phase was dried over anhydrous sodium sulfate, concentrated, and purified by column chromatography (V Petroleum ether /V Acetic acid ethyl ester =3/1) to give 357mg of white solid in 83% yield. UPLC-MS calculated for C 34 H 40 ClN 7 O 4 [M+H] + :646.28,found:646.29.UPLC-retention time:6.7min. 1 H NMR(400MHz,Chloroform-d)δ8.78(s,2H),7.72(s,1H),7.61(s,1H),7.56(d,J＝8.8Hz,1H),7.47(d,J＝10.9Hz,1H),6.99–6.91(m,2H),6.82(dd,J＝8.7,2.4Hz,1H).4.87(d,J＝13.1Hz,2H),3.44-3.48(m，4H),2.94(t,J＝12.8Hz,2H),2.40-2.31(m,4H),2.14(s,3H),1.90-1.88(m,4H),1.45(s,9H),1.33–1.06(m,4H).
Figure BDA0003259005860000321
Intermediate 27 (200 mg,1.0 eq.) was dissolved in 5mL dichloromethane, added with 1mL dioxane hydrochloride, stirred overnight at room temperature, TLC detected complete reaction of the starting material, filtered, the solid dissolved in 10mL water, pH adjusted to 8-9 with aqueous ammonia, extracted with dichloromethane, dried over anhydrous magnesium sulfate, filtered and concentrated to give 162mg white solid in 96% yield. UPLC-MS calculated for C 29 H 32 ClN 7 O 2 [M+H] + :546.23,found:545.95.UPLC-retention time:4.5min.
Figure BDA0003259005860000322
Intermediate 28 (162 mg,1.0 eq.) and intermediate 15 (98 mg,1.2 eq.) were dissolved in 5mL anhydrous N, N-dimethylformamide, protected with argon, N-diisopropylethylamine (104 μl,2.0 eq.) was added, and reacted overnight at 90 ℃ with TLC detection, complete reaction of starting material, concentrated reaction solution, purified by column chromatography (V Dichloromethane (dichloromethane) /V Methanol =80/1-30/1), 169mg of fluorescent green solid was obtained in 71% yield. UPLC-MS calculated for C 42 H 40 ClN 9 O 6 [M+H] + :802.28found:802.03.UPLC-retention time:5.6min. 1 H NMR(400MHz,DMSO-d 6 )δ11.12(s,1H),10.18(s,1H),8.90(s,2H),7.94(d,J＝9.0Hz,1H),7.78(d,J＝10.4Hz,2H),7.68(d,J＝10.0Hz,1H),7.51(s,1H),7.39(t,J＝9.6Hz,1H),7.20(s,1H),7.12(d,J＝8.6Hz,1H),6.97–6.91(m,1H),4.78(d,J＝13.0Hz,2H),4.23(d,J＝13.5Hz,2H),3.64(d,J＝12.0Hz,2H),3.35(t,J＝13.0Hz,2H),3.25–3.01(m,4H),2.93-8.84(m,2H),2.34-2.17(m,2H),2.11(s,3H),2.03(d,J＝12.1Hz,1H),1.89(d,J＝12.7Hz,2H),1.26-1.12(m,4H).
Figure BDA0003259005860000323
Intermediate 24 (1.8 g,1.0 eq.) was dissolved in 40mL anhydrous N, N-dimethylformamide, 60% nah (0.5 g,1.5 eq.) was added under ice-bath conditions, after stirring for 0.5h, starting material 10 (1.5 g,1.2 eq.) was added, the reaction was carried out at room temperature overnight, TLC detection, starting material reaction was complete, 60mL water was added to the reaction solution, extraction was performed with ethyl acetate, the organic phase was added to anhydrous sodium sulfate, dried and concentrated, and purified by column chromatography (V Petroleum ether /V Acetic acid ethyl ester =10/1-3/1) to give 2.7g of white solid in 94% yield. UPLC-MS calculated for C 19 H 19 ClN 2 O 3 [M+H] + :359.11,found:359.12.UPLC-retention time:6.9min. 1 H NMR(400MHz,CDCl 3 )δ7.56(t,J＝8.7Hz,1H),7.38(s,1H),7.18(d,J＝8.5Hz,1H),6.91(d,J＝7.5Hz,2H),6.80(d,J＝9.1Hz,1H),2.11(s,3H)1.52(s,9H).
Figure BDA0003259005860000331
Intermediate 25 (2.7 g,1.0 eq.) was dissolved in 60mL dichloromethane, added with dioxane hydrochloride 20mL and stirred overnight at room temperature, TLC detected complete reaction of starting material and filtered to give 2.2g of white solid in 97% yield. UPLC-MS calculated for C 14 H 11 ClN 2 O[M+H] + :259.06,found:258.95.UPLC-retention time:3.5min.
Example 4:
Figure BDA0003259005860000332
intermediate 28 (66 mg,1.0 eq.) and intermediate 16 (43 mg,1.2 eq.) were dissolved in 5mL anhydrous N, N-dimethylformamide under argon, and addedN, N-diisopropylethylamine (44. Mu.L, 2.0 eq.) was reacted at 90℃for 5h, the reaction mixture was concentrated and purified by column chromatography (V Dichloromethane (dichloromethane) /V Methanol =80/1-30/1), 79mg of fluorescent green solid was obtained in 80% yield. UPLC-MS calculated for C 42 H 39 ClFN 9 O 6 [M+H] + :820.27found:819.99.UPLC-retention time:5.6min. 1 H NMR(400MHz,DMSO-d 6 )δ11.10(s,1H),10.14(s,1H),8.87(s,2H),7.90(d,J＝9.9Hz,1H),7.80(d,J＝11.0Hz,1H),7.72(s,1H),7.64(d,J＝9.3Hz,1H),7.58(d,J＝7.8Hz,1H),7.16(s,1H),7.08(d,J＝8.7Hz,1H),6.91(d,J＝10.2Hz,1H),5.09(d,J＝11.7Hz,1H),4.75(d,J＝13.2Hz,2H),3.76(d,J＝13.2Hz,2H),3.61(d,J＝10.9Hz,2H),3.38–3.19(m,4H),3.10(s,2H),3.02(t,J＝12.5Hz,2H),2.86(s,1H),2.57(dd,J＝34.6,17.0Hz,2H),2.19(s,1H),2.07(s,3H),2.03(s,1H),1.85(d,J＝12.7Hz,2H),1.18(d,J＝12.4Hz,2H).
Example 5:
Figure BDA0003259005860000341
raw material 29 (127 mg,1.2 eq.) and HATU (340 mg,1.2 eq.) were dissolved in 15mL of anhydrous N, N-dimethylformamide under argon, N-diisopropylethylamine (390 μl,3.0 eq.) was added, stirred at room temperature for 0.5h, intermediate 26 (220 mg,1.0 eq.) was added, reaction was carried out for 6h, tlc detection, raw material reaction was complete, 20mL of water was added to the reaction solution, extraction was carried out with ethyl acetate, the organic phase was added to anhydrous sodium sulfate, dried, concentrated, and purified by column chromatography (V Petroleum ether /V Acetic acid ethyl ester =3/1) to give 250mg of a white solid in 87% yield. UPLC-MS calculated for C 20 H 13 ClFN 3 O 2 [M+H] + :382.07,found:381.89.UPLC-retention time:5.8min. 1 H NMR(400MHz,DMSO-d 6 )δ10.49(s,1H),8.78(s,1H),8.46(t,J＝9.3Hz,1H),7.90(d,J＝9.1Hz,1H),7.76(s,1H),7.66(d,J＝9.0Hz,1H),7.35(d,J＝8.6Hz,1H),7.18(s,1H),7.11(d,J＝8.8Hz,1H),6.91(d,J＝8.7Hz,1H),2.09(s,3H).
Figure BDA0003259005860000342
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Intermediate 30 (236 mg,1.0 eq.), raw material 2 (127 mg,1.1 eq.), potassium carbonate (171 mg,2.0 eq.) were dissolved in 10mL of anhydrous N, N-dimethylformamide, and reacted overnight at 85 ℃ under argon, the reaction was detected by TLC, 20mL of water was added to the reaction solution, extracted with ethyl acetate, and the organic phase was dried over anhydrous sodium sulfate, concentrated, purified by column chromatography (V Petroleum ether /V Acetic acid ethyl ester =3/1) to give 250mg of a white solid in 87% yield. UPLC-MS calculated for C 29 H 30 ClN 5 O 4 [M+H] + :548.20,found:547.87.UPLC-retention time:6.9min. 1 H NMR(400MHz,DMSO-d 6 )δ10.06(s,1H),8.71(s,1H),8.06(dd,J＝9.0,2.4Hz,1H),7.90(d,J＝8.7Hz,1H),7.76(d,J＝2.2Hz,1H),7.66(dd,J＝8.8,2.4Hz,1H),7.17(d,J＝2.4Hz,1H),7.07(d,J＝8.7Hz,1H),6.90(dd,J＝8.8,2.6Hz,2H),3.66–3.58(m,4H),3.45–3.36(m,4H),2.07(s,3H),1.39(s,9H).
Figure BDA0003259005860000351
Intermediate 31 (200 mg,1.0 eq.) was dissolved in 5mL dichloromethane, 1mL dioxane hydrochloride was added, stirred overnight at room temperature, TLC detected complete reaction of the starting material, filtered, the solid dissolved in 10mL water, pH adjusted to 8-9 with aqueous ammonia, extracted with dichloromethane, dried over anhydrous magnesium sulfate, filtered and concentrated to give 157mg white solid in 96% yield. UPLC-MS calculated for C 24 H 22 ClN 5 O 2 [M+H] + :448.15,found:447.97.UPLC-retention time:4.9min.
Figure BDA0003259005860000352
Intermediate 32 (100 mg,1.0 eq.) and intermediate 5 (57 mg,1.2 eq.) were dissolved in 10ml 1, 2-dichloroethane, 1 drop acetic acid was added, and after stirring at room temperature for 0.5h, sodium triacetylborohydride (95 mg) was added2.0 eq.) overnight at room temperature, TLC detected complete reaction of starting material, quenched by addition of 5mL methanol, concentrated, purified by column chromatography (V Dichloromethane (dichloromethane) /V Methanol =20/1) to yield 122mg of white solid in 85% yield. UPLC-MS calculated for C 35 H 41 ClN 6 O 4 [M+H] + :645.29,found:644.95.UPLC-retention time:6.0min.
Figure BDA0003259005860000353
Intermediate 33 (100 mg,1.0 eq.) was dissolved in 5mL dichloromethane, 1mL dioxane hydrochloride was added, stirred overnight at room temperature, TLC detected complete reaction of the starting material, filtered, the solid dissolved in 10mL water, pH adjusted to 8-9 with aqueous ammonia, extracted with dichloromethane, dried over anhydrous magnesium sulfate, filtered and concentrated to give 82mg white solid in 97% yield. UPLC-MS calculated for C 30 H 33 ClN 6 O 2 [M+H] + :545.24,found:545.25.UPLC-retention time:3.8min.
Figure BDA0003259005860000354
Intermediate 28 (66 mg,1.0 eq.) and intermediate 15 (40 mg,1.2 eq.) were dissolved in 5mL anhydrous N, N-dimethylformamide, protected with argon, N-diisopropylethylamine (42 μl,2.0 eq.) was added, the reaction was allowed to proceed to 90 ℃ for 5h, tlc detected complete reaction of starting materials, the reaction concentrated, and purified by column chromatography (V Dichloromethane (dichloromethane) /V Methanol =80/1-30/1), 68mg of fluorescent green solid was obtained in 70% yield. UPLC-MS calculated for C 43 H 41 ClN 8 O 6 [M+H] + :801.28found:801.29.UPLC-retention time:5.2min. 1 H NMR(400MHz,DMSO-d 6 )δ11.06(s,1H),10.14(s,1H),8.77(s,1H),8.21–8.05(m,1H),7.91(d,J＝9.1Hz,1H),7.79–7.62(m,3H),7.36–6.88(m,5H),5.05(s,1H),4.51(d,J＝14.0Hz,2H),4.08(d,J＝13.1Hz,2H),3.61(d,J＝11.9Hz,2H),3.36(s,2H),3.02(d,J＝34.8Hz,5H),2.71–2.50(m,4H),2.07(m,8H),1.27–1.25(m,2H).
Example 6:
Figure BDA0003259005860000361
intermediate 28 (66 mg,1.0 eq.) and intermediate 17 (39 mg,1.2 eq.) were dissolved in 5mL anhydrous N, N-dimethylformamide, protected with argon, N-diisopropylethylamine (42 μl,2.0 eq.) was added, the reaction was warmed to 90 ℃ and allowed to react for 5h, tlc detected complete starting material reaction, the reaction concentrated, and purified by column chromatography (V Dichloromethane (dichloromethane) /V Methanol =80/1-30/1), to give 82mg of fluorescent green solid in 83% yield. UPLC-MS calculated for C 42 H 39 ClFN 9 O 6 [M+H] + :819.27found:819.15.UPLC-retention time:5.4min. 1 H NMR(400MHz,DMSO-d 6 )δ11.09(s,1H),10.13(s,1H),8.83–8.70(m,1H),8.21–8.08(m,1H),7.91(d,J＝8.7Hz,1H),7.77–7.64(m,3H),7.45(d,J＝7.4Hz,1H),7.16(d,J＝2.4Hz,1H),7.07(dd,J＝13.7,8.9Hz,2H),6.91(dd,J＝8.8,2.4Hz,1H),5.08(dd,J＝12.8,5.4Hz,1H),4.52(d,J＝13.4Hz,3H),3.62(d,J＝10.7Hz,5H),3.33(t,J＝13.0Hz,2H),2.97–2.53(m,6H),2.14–1.81(m,8H),1.40–1.38(m,2H).
Example 7:
Figure BDA0003259005860000362
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Figure BDA0003259005860000371
raw material 35 (150 mg,1.0 eq.), intermediate 20 (383 mg,1.1 eq.) and potassium carbonate (340 mg,2.0 eq.) were dissolved in 12mL anhydrous N, N-dimethylformamide, and reacted overnight at 85 ℃ under argon protection, TLC was detected, the raw material was reacted completely, 20mL of water was added to the reaction solution, extracted with ethyl acetate, and the organic phase was dried over anhydrous sodium sulfate, concentrated, purified by column chromatography (V Petroleum ether /V Acetic acid ethyl ester =3/1-1/1) to give 429mg of white solid in 91% yield.UPLC-MS calculated for C 22 H 32 N 4 O 2 [M+H] + :385.25,found:385.05.UPLC-retention time:3.5min. 1 H NMR(400MHz,Chloroform-d)δ7.47(d,J＝8.7Hz,2H),6.84(d,J＝8.8Hz,2H),3.85(m,2H),3.42(m,4H),2.85(m,2H),2.35(m,4H),2.20(m,2H),1.91–1.67(m,3H),1.46(s,9H),1.31–1.19(m,2H).
Figure BDA0003259005860000372
Intermediate 36 (200 mg,1.0 eq.) was dissolved in 10mL ethanol/water (V Ethanol /V Water and its preparation method In the mixed solution of=5/1), the temperature is raised and the reflux is carried out overnight, TLC detection is carried out, the raw materials are reacted completely, the reaction solution is concentrated, 5mL of water is added to dissolve solid, the pH is regulated to about 4.5 by 1M hydrochloric acid, ethyl acetate is used for extraction, and the organic phase is added to be dried by anhydrous sodium sulfate and then concentrated, so that white solid 195mg is obtained, and the yield is 93%. UPLC-MS calculated for C 22 H 33 N 3 O 4 [M+H] + :404.25,found:404.26.UPLC-retention time:4.0min.
Figure BDA0003259005860000381
Intermediate 37 (100 mg,1.0 eq.) and HATU (113 mg,1.2 eq.) were dissolved in 5mL of anhydrous N, N-dimethylformamide under argon, N-diisopropylethylamine (130 μl,2.0 eq.) was added, stirred at room temperature for 0.5h, intermediate 26 (81 mg,1.1 eq.) was added, reaction was carried out for 6h, tlc detection, starting material reaction was complete, 20mL of water was added to the reaction solution, extracted with ethyl acetate, the organic phase was dried over anhydrous sodium sulfate, concentrated, and purified by column chromatography (V Petroleum ether /V Acetic acid ethyl ester =3/1) to give 136mg of a white solid in 85% yield. UPLC-MS calculated for C 36 H 42 ClN 5 O 4 [M+H] + :644.29,found:644.30.UPLC-retention time:6.7min.
Figure BDA0003259005860000382
Intermediate 38 (100 mg,1.0 eq.) was dissolved in 5mL dichloromethane, added 1mL dioxane hydrochloride, stirred overnight at room temperature, TLC detected complete reaction of the starting material, filtered, the solid dissolved in 10mL water, pH adjusted to 8-9 with aqueous ammonia, extracted with dichloromethane, dried over anhydrous magnesium sulfate, filtered and concentrated to give 81mg white solid in 96% yield. UPLC-MS calculated for C 31 H 34 ClN 5 O 2 [M+H] + :544.24,found:544.25.UPLC-retention time:4.0min.
Figure BDA0003259005860000383
Intermediate 39 (50 mg,1.0 eq.) and intermediate 17 (30 mg,1.2 eq.) were dissolved in 5mL anhydrous N, N-dimethylformamide, protected with argon, N-diisopropylethylamine (32 μl,2.0 eq.) was added, the reaction was warmed to 90 ℃ and allowed to react for 5h, tlc detected complete starting material reaction, the reaction concentrated, and purified by column chromatography (V Dichloromethane (dichloromethane) /V Methanol =80/1-30/1), 54mg of fluorescent green solid was obtained in 73% yield. UPLC-MS calculated for C 44 H 42 ClN 7 O 6 [M+H] + :800.29found:800.30.UPLC-retention time:5.3min. 1 H NMR(400MHz,DMSO-d 6 )δ11.09(s,1H),10.06(s,1H),7.91(d,J＝8.0Hz,3H),7.79(s,1H),7.72-7.68(m,2H),7.45(d,J＝8.0Hz,1H),7.16(s,1H),7.11-7.06(m,3H),6.91(d,J＝8.0Hz,1H),5.08(dd,J＝8.0Hz,4Hz,1H),3.99(s,2H),3.61(s,6H),3.16-3.13(m,6H),2.96-2.83(m,3H),2.07(s,3H),2.01(s,2H),1.88(d,J＝8.0Hz,2H),1.43-1.35(m,2H).
Example 8:
Figure BDA0003259005860000391
intermediate 39 (50 mg,1.0 eq.) and intermediate 17 (30 mg,1.2 eq.) were dissolved in 5mL anhydrous N, N-dimethylformamide, protected with argon, N-diisopropylethylamine (32 μl,2.0 eq.) was added, the reaction was allowed to proceed to 90℃for 5h, TLC detection of complete starting material reaction was performed, and the reaction solution was concentratedCondensation, column chromatography purification (V Dichloromethane (dichloromethane) /V Methanol =80/1-30/1), 64mg of fluorescent green solid was obtained in 85% yield. UPLC-MS calculated for C 44 H 41 ClFN 7 O 6 [M+H] + :818.28found:818.29.UPLC-retention time:5.3min.UPLC-MS calculated for C 42 H 39 ClFN 9 O 6 [M+H] + :819.27found:819.15.UPLC-retention time:5.4min. 1 H NMR(400MHz,DMSO-d 6 )δ11.09(s,1H),10.06(s,1H),7.94-7.90(m,3H),7.79(s,1H),7.72-7.68(m,2H),7.45(d,J＝8.0Hz,1H),7.16(s,1H),7.11-7.06(m,3H),6.91(d,J＝8.0Hz,1H),5.08(dd,J＝8.0Hz,4Hz,1H),3.99(s,2H),3.61(s,6H),3.16-3.13(m,6H),2.96-2.83(m,3H),2.07-2.01(m,4H),1.96(d,J＝8.0Hz,2H),1.40-1.35(m,2H).
Example 9:
Figure BDA0003259005860000392
Figure BDA0003259005860000401
raw material 40 (1.0 g,1.0 eq.) was dissolved in 20mL of anhydrous N, N-dimethylformamide, 60% nah (0.3 g,1.5 eq.) was added under ice-bath conditions, after stirring for 0.5h, raw material 10 (0.9 g,1.2 eq.) was added, reacted overnight at room temperature, TLC detection was completed, 30mL of water was added to the reaction solution, extracted with ethyl acetate, the organic phase was added to anhydrous sodium sulfate, dried and concentrated, and purified by column chromatography (V Petroleum ether /V Acetic acid ethyl ester =10/1-3/1) to give 1.5g of white solid in 94% yield. UPLC-MS calculated for C 18 H 23 ClN 2 O 3 [M+H] + :351.14,found:351.15.UPLC-retention time:6.9min.
Figure BDA0003259005860000402
Intermediate 41 (1.5 g,1.0 eq.) was dissolved in 30mL of dichloromethane,dioxahexacyclic hydrochloride (10 mL) is added, stirred at room temperature overnight, TLC (thin layer chromatography) detects complete reaction of raw materials, and white solid (146 mg) is obtained through filtration, and the yield is 96%. UPLC-MS calculated for C 13 H 15 ClN 2 O[M+H] + :251.09,found:250.94.UPLC-retention time:2.0min.
Figure BDA0003259005860000403
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Intermediate 23 (140 mg,1.0 eq.) and HATU (158 mg,1.2 eq.) were dissolved in 10mL of anhydrous N, N-dimethylformamide under argon, N-diisopropylethylamine (270 μl,3.0 eq.) were added, stirred at room temperature for 0.5h, intermediate 42 (109 mg,1.0 eq.) was added, reaction was carried out for 6h, tlc detection, starting material reaction was complete, 20mL of water was added to the reaction solution, extracted with ethyl acetate, the organic phase was added to anhydrous sodium sulfate, dried, concentrated, and purified by column chromatography (V Petroleum ether /V Acetic acid ethyl ester =3/1) to yield 185mg of white solid in 84% yield. UPLC-MS calculated for C 33 H 44 ClN 7 O 4 [M+H] + :638.31,found:638.10.UPLC-retention time:4.2min. 1 H NMR(400MHz,DMSO-d 6 )δ8.70(s,2H),8.08(d,J＝7.4Hz,1H),7.83(d,J＝8.8Hz,1H),7.36(d,J＝2.3Hz,1H),7.11(dd,J＝8.8,2.3Hz,1H),4.72–4.62(m,2H),4.56–4.47(m,1H),3.76(s,1H),2.89(t,J＝11.8Hz,2H),2.27(m,5H),2.14–2.00(m,4H),1.90–1.73(m,4H),1.46(t,J＝9.8Hz,4H),1.35(s,9H),1.22-1.20(m,4H),1.07–0.94(m,2H).
Figure BDA0003259005860000411
Intermediate 43 (126 mg,1.0 eq.) was dissolved in 5mL dichloromethane, dioxane hydrochloride 1mL was added, stirred overnight at room temperature, TLC detected complete reaction of the starting material, filtered, the solid dissolved in 10mL water, pH adjusted to 8-9 with aqueous ammonia, extracted with dichloromethane, dried over anhydrous magnesium sulfate, filtered and concentrated to a white solid 102mg, 96% yield. UPLC-MS calculated for C 28 H 36 ClN 7 O 2 [M+H] + :538.26,found:537.96.UPLC-retention time:5.2min.
Figure BDA0003259005860000412
Intermediate 44 (50 mg,1.0 eq.) and intermediate 17 (30 mg,1.2 eq.) were dissolved in 5mL anhydrous N, N-dimethylformamide, protected with argon, N-diisopropylethylamine (32 μl,2.0 eq.) was added, the reaction was warmed to 90 ℃ and allowed to react for 5h, tlc detected complete starting material reaction, the reaction concentrated, and purified by column chromatography (V Dichloromethane (dichloromethane) /V Methanol =80/1-30/1), 63mg of fluorescent green solid was obtained in 84% yield. UPLC-MS calculated for C 41 H 43 ClFN 9 O 6 [M+H] + :812.30found:812.28.UPLC-retention time:4.5min. 1 H NMR(400MHz,DMSO-d 6 )δ11.10(s,1H),8.73(s,2H),8.13(d,J＝7.4Hz,1H),7.89–7.74(m,2H),7.58(d,J＝7.3Hz,1H),7.35(d,J＝2.3Hz,1H),7.11(dd,J＝8.8,2.4Hz,1H),5.10(dd,J＝12.8,5.3Hz,1H),4.70(d,J＝12.9Hz,2H),4.51(s,2H),3.68(m,6H),3.34(t,J＝11.9Hz,2H),3.04–2.76(m,4H),1.94(m,8H),1.47(t,J＝10.3Hz,5H),1.23–1.09(m,2H).
Example 10:
Figure BDA0003259005860000421
intermediate 45 (200 mg,1.0 eq.) and HATU (575 mg,1.2 eq.) were dissolved in 15mL of anhydrous N, N-dimethylformamide under argon, N-diisopropylethylamine (660 μl,3.0 eq.) was added, stirred at room temperature for 0.5h, intermediate 42 (399 mg,1.1 eq.) was added, reaction was performed for 6h, tlc detection, starting material reaction was complete, 20mL of water was added to the reaction solution, extraction was performed with ethyl acetate, the organic phase was added to anhydrous sodium sulfate, dried, concentrated, and purified by column chromatography (V) Petroleum ether /V Acetic acid ethyl ester =3/1) to yield 419mg of white solid in 85% yield. UPLC-MS calculated for C 18 H 16 Cl 2 N 4 O 2 [M+H] + :391.07,found:390.87.UPLC-retention time:4.9min. 1 H NMR(400MHz,Chloroform-d)δ8.29(d,J＝8.6Hz,1H),7.98(d,J＝8.4Hz,1H),7.71(d,J＝8.9Hz,1H),7.58(d,J＝8.7Hz,1H),7.01(s,1H),6.86(d,J＝10.5Hz,1H),4.35(s,1H),4.10(d,J＝11.3Hz,1H),2.32–2.13(m,4H),1.79–1.48(m,4H).
Figure BDA0003259005860000431
Intermediate 46 (340 mg,1.0 eq.), intermediate 20 (295 mg,1.2 eq.), potassium carbonate (240 mg,2.0 eq.) were dissolved in 10mL anhydrous N, N-dimethylformamide, protected with argon, reacted overnight at 85 ℃, TLC detected that the starting material was complete, 20mL water was added to the reaction solution, extracted with ethyl acetate, the organic phase was dried over anhydrous sodium sulfate, concentrated, purified by column chromatography (V Petroleum ether /V Acetic acid ethyl ester =3/1-1/1) to give 429mg of white solid in 91% yield. UPLC-MS calculated for C 33 H 44 ClN 7 O 4 [M+H] + :638.31,found:638.04.UPLC-retention time:4.3min. 1 H NMR(400MHz,DMSO-d 6 )δ8.57(d,J＝8.2Hz,1H),7.82(d,J＝8.8Hz,1H),7.76(d,J＝9.6Hz,1H),7.35(d,J＝2.3Hz,1H),7.29(d,J＝9.7Hz,1H),7.10(dd,J＝8.8,2.4Hz,1H),4.55–4.33(m,3H),3.81(tt,J＝11.1,8.2,5.8Hz,1H),2.95(t,J＝11.9Hz,2H),2.33–2.18(m,4H),2.15–2.00(m,4H),2.00–1.71(m,6H),1.53(dq,J＝51.3,11.9Hz,5H),1.35(s,9H),1.25–0.99(m,4H).
Figure BDA0003259005860000432
Intermediate 11 (200 mg,1.0 eq.) was dissolved in 5mL dichloromethane, added with 1mL dioxane hydrochloride, stirred overnight at room temperature, TLC detected complete reaction of the starting material, filtered, the solid dissolved in 10mL water, pH adjusted to 8-9 with aqueous ammonia, extracted with dichloromethane, dried over anhydrous magnesium sulfate, filtered and concentrated to yield 162mg white solid in 96%. UPLC-MS calculated for C 28 H 36 ClN 7 O 2 [M+H] + :538.26,found:538.27.UPLC-retention time:4.0min.
Figure BDA0003259005860000433
Intermediate 48 (90 mg,1.0 eq.) and intermediate 17 (55 mg,1.2 eq.) were dissolved in 5mL anhydrous N, N-dimethylformamide, protected with argon, N-diisopropylethylamine (60 μl,2.0 eq.) was added, heated to 90 ℃ and reacted for 5h, tlc detected complete reaction of starting material, reaction concentrated, purified by column chromatography (V Dichloromethane (dichloromethane) /V Methanol =80/1-30/1), to give 82mg of fluorescent green solid in 83% yield. UPLC-MS calculated for C 41 H 44 ClN 9 O 6 [M+H] + :794.31found:794.17.
UPLC-retention time:4.6min. 1 H NMR(400MHz,DMSO-d 6 )δ11.08(s,1H),8.58(d,J＝8.2Hz,1H),7.85–7.70(m,3H),7.50–7.29(m,4H),7.10(d,J＝11.2Hz,1H),5.06(dd,J＝12.8,5.4Hz,1H),4.49(t,J＝12.3Hz,3H),4.19(dd,J＝8.9,5.6Hz,2H),3.89–3.78(m,1H),3.66–3.53(m,2H),3.22–2.77(m,8H),2.34–1.78(m,10H),1.67–1.39(m,5H),1.27–1.14(m,2H).
Example 11:
Figure BDA0003259005860000441
raw material 10 (5.0 g,1.0 eq.), raw material 48 (5.5 g,1.0 eq.), potassium carbonate (6.7 g,1.5 eq.) were dissolved in 10mL anhydrous N, N-dimethylformamide under argon protection, and the temperature was raised to 90 ℃ for reaction overnight, TLC detection of the raw material reaction was complete, the reaction solution was concentrated, and column chromatography was performed (V Petroleum ether /V Acetic acid ethyl ester =10/1) to give 4.7g of a white solid, yield 48%. UPLC-MS calculated for C 15 H 11 ClN 2 O 3 [M+H] + :304.04,found:303.95.UPLC-retention time:6.3min. 1 H NMR(400MHz,DMSO-d 6 )δ8.13(s,2H),7.91(d,J＝8.0Hz,1H),7.26(s,1H),6.90(dd,J＝8.0Hz,2.4Hz,1H),2.13(s,6H).
Figure BDA0003259005860000442
Intermediate 49 (4.7 g,1.0 eq.) was dissolved in 50mL methanol, reduced iron powder (2.6 g,3.0 eq.) and 10mL HCl were added, the temperature was raised to 50 ℃ for 4h, tlc detected complete reaction of starting material, celite suction filtration, filtrate concentration, pH adjustment to neutral using 3N NaOH, solvent spin drying, column chromatography purification (V Petroleum ether /V Acetic acid ethyl ester =4/1) to give 4.7g of an off-white solid in 48% yield. UPLC-MS calculated for C 15 H 13 ClN 2 O[M+H] + :273.08,found:272.95.UPLC-retention time:3.0min. 1 H NMR(400MHz,DMSO-d 6 )δ7.85(d,J＝8.0Hz,1H),7.03(s,1H),6.80(d,J＝8.0Hz,1H),6.47(s,1H),6.31(s,1H),4.99(s,2H),2.13(s,6H).
Figure BDA0003259005860000451
Intermediate 7 (400 mg,1.0 eq.), intermediate 50 (324 mg,1.2 eq.), HATU (375 mg,1.2 eq.) were dissolved in 10mL anhydrous N, N-dimethylformamide, protected with argon, N-diisopropylethylamine (520 μl,3.0 eq.) was added, reacted overnight at room temperature, TLC detection, starting material reaction was complete, the reaction solution was added with 20mL water, extracted with ethyl acetate, the organic phase was dried over anhydrous sodium sulfate, concentrated, and purified by column chromatography (V Petroleum ether /V Acetic acid ethyl ester =2/1) to give 550mg of a white solid in 84% yield. UPLC-MS calculated for C 35 H 42 ClN 7 O 4 [M+H] + :660.30,found:660.09.UPLC-retention time:5.7min. 1 H NMR(400MHz,DMSO-d 6 )δ10.03(s,1H),8.84(s,2H),7.88(d,J＝8.0Hz,2H),7.83(s,2H),7.12(s,1H),6.82(d,J＝8.0Hz,4H),3.90(d,J＝8.0Hz,2H),3.80(s,4H),2.37(s,4H),2.13(d,J＝4.0Hz,2H),2.01(s,6H),1.88-1.85(s,3H),1.35(s,2H),1.19(s,9H),0.95-0.88(m,2H).
Figure BDA0003259005860000452
Intermediate 51 (460 mg,1.0 eq.) was dissolved in 20mL dichloromethane and 5mL 4M hydrochloric acid was addedThe dioxane solution is stirred overnight at room temperature, TLC detection is carried out, the raw materials are completely reacted, the filtration is carried out, the solid is dissolved in 5mL of water, the pH is adjusted to 8-9 by ammonia water, the dichloromethane extraction is carried out, anhydrous magnesium sulfate is added for drying, 355mg of white solid is obtained by filtration and concentration, and the yield is 91%. UPLC-MS calculated for C 30 H 34 ClN 7 O 2 [M+H] + :560.25,found:560.02.UPLC-retention time:3.1min.
Figure BDA0003259005860000453
Intermediate 90 (100 mg,1.0 eq.) and intermediate 15 (60 mg,1.2 eq.) were dissolved in 5mL anhydrous N, N-dimethylformamide, protected with argon, N-diisopropylethylamine (62 μl,2.0 eq.) was added, the reaction was allowed to proceed to 90 ℃ for 8h, tlc detected complete reaction of starting materials, the reaction concentrated, and purified by column chromatography (V Dichloromethane (dichloromethane) /V Methanol =80/1-30/1), 90mg of fluorescent green solid was obtained in 62% yield. UPLC-MS calculated for C 43 H 42 ClN 9 O 6 [M+H] + :816.30,found:815.99.UPLC-retention time:5.0min. 1 H NMR(400MHz,DMSO-d 6 )δ11.06(s,1H),10.18(s,1H),8.96(s,2H),7.89(d,J＝8.0Hz,1H),7.65(d,J＝8.0Hz,1H),7.67(s,2H),7.34(s,1H),7.25(d,J＝8.0Hz,1H),7.11(s,1H),6.83(dd,J＝8.0Hz,4.0Hz,1H),5.06-5.02(m,1H),4.79-4.76(m,2H),4.10-4.07(m,2H),3.60(s,4H),3.49-3.43(m,2H),3.06(s,4H),3.00-2.94(m,2H),2.89-2.82(m,1H),2.15(s,1H),2.02(s,6H),1.83(d,J＝12.0Hz,2H),1.27(d,J＝8.0Hz,2H),1.19(s,1H).
Example 12:
Figure BDA0003259005860000461
intermediate 90 (50 mg,1.0 eq.) and intermediate 15 (32 mg,1.2 eq.) were dissolved in 5mL anhydrous N, N-dimethylformamide, protected with argon, N-diisopropylethylamine (31 μl,2.0 eq.) was added, the reaction was allowed to proceed to 90 ℃ for 4h, tlc detected complete reaction of starting materials, the reaction concentrated, and purified by column chromatography (V Dichloromethane (dichloromethane) /V Methanol =80/1-30/1), 59mg of fluorescent green solid was obtained in 79% yield. UPLC-MS calculated for C 43 H 41 ClFN 9 O 6 [M+H] + :834.29,found:834.06.UPLC-retention time:6.5min. 1 H NMR(400MHz,DMSO-d 6 )δ11.09(s,1H),10.20(s,1H),8.96(s,2H),7.89(d,J＝8.0Hz,1H),7.70(d,J＝8.0Hz,1H),7.67(s,2H),7.34(s,1H),7.11(d,J＝2.0Hz,1H),6.83(dd,J＝8.0Hz,4.0Hz,1H),5.10-5.06(m,1H),4.80-4.76(m,2H),4.09(m,6H),3.64-3.61(m,4H),3.09(s,4H),2.93-2.85(m,3H),2.01(s,6H),1.89-1.86(m,2H),1.39(q,J＝12.0Hz,2H).
Example 13:
Figure BDA0003259005860000462
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Figure BDA0003259005860000471
intermediate 23 (400 mg,1.0 eq.), intermediate 50 (323 mg,1.2 eq.), HATU (560 mg,1.2 eq.) were dissolved in 10mL anhydrous N, N-dimethylformamide, protected with argon, N-diisopropylethylamine (515 μl,3.0 eq.) was added, reacted overnight at room temperature, TLC detection, starting material reaction was complete, the reaction solution was added with 20mL water, extracted with ethyl acetate, the organic phase was dried over anhydrous sodium sulfate, concentrated, and purified by column chromatography (V Petroleum ether /V Acetic acid ethyl ester =3/1) to give 550mg of a white solid in 85% yield. UPLC-MS calculated for C 35 H 42 ClN 7 O 4 [M+H] + :660.30,found:660.09.UPLC-retention time:5.7min. 1 H NMR(400MHz,DMSO-d 6 )δ10.03(s,1H),8.83(s,2H),7.88(d,J＝8.0Hz,1H),7.54(s,2H),7.11(s,1H),6.82(d,J＝8.0Hz,1H),4.92(d,J＝12.0Hz,2H),3.29(s,2H),2.27(s,2H),2.12(d,J＝8.0Hz,2H),2.01(s,6H),1.81-1.75(m,3H),1.35(s,9H),1.18(m,2H),1.01-0.98(m,2H).
Figure BDA0003259005860000472
Intermediate 53 (500 mg,1.0 eq.) was dissolved in 20mL dichloromethane, 5mL 4m dioxane hydrochloride solution was added, stirred overnight at room temperature, TLC detection, complete reaction of the starting material, filtration, dissolution of the solid in 5mL water, pH adjustment to 8-9 with aqueous ammonia, extraction with dichloromethane, drying with anhydrous magnesium sulfate, filtration and concentration to give 400mg white solid with 94% yield. UPLC-MS calculated for C 30 H 34 ClN 7 O 2 [M+H] + :560.25,found:560.02.UPLC-retention time:3.2min.
Figure BDA0003259005860000481
Intermediate 90 (100 mg,1.0 eq.) and intermediate 15 (60 mg,1.2 eq.) were dissolved in 5mL anhydrous N, N-dimethylformamide, protected with argon, N-diisopropylethylamine (62 μl,2.0 eq.) was added, the reaction was allowed to proceed to 90 ℃ for 8h, tlc detected complete reaction of starting materials, the reaction concentrated, and purified by column chromatography (V Dichloromethane (dichloromethane) /V Methanol =80/1-30/1), 85mg of fluorescent green solid was obtained in 58% yield. UPLC-MS calculated for C 43 H 42 ClN 9 O 6 [M+H] + :816.30,found:816.07.UPLC-retention time:5.0min. 1 H NMR(400MHz,DMSO-d 6 )δ11.07(s,1H),10.12(s,1H),8.87(s,2H),7.89(d,J＝8.0Hz,1H),7.73(d,J＝8.0Hz,1H),7.67(s,2H),7.46(s,1H),7.33(d,J＝8.0Hz,1H),7.12(d,J＝4.0Hz,1H),6.83(dd,J＝6.4Hz,2.0Hz,1H),5.10-5.04(m,1H),4.76-4.72(m,2H),4.19-4.16(m,2H),3.66-3.42(m,8H),3.13-2.99(m,4H),2.89-2.81(m,1H),2.20(s,1H),2.01(s,6H),1.92-1.89(m,2H),1.39(m,3H).
Example 14:
Figure BDA0003259005860000482
intermediate 90 (50 mg,1.0 eq.) and intermediate 15 (32 mg,1.2 eq.) were dissolved in 5mL anhydrous N, N-dimethylformamide under argon, N-diisopropylethylamine (31. Mu.L, 2.0 eq.) was added and warmed to 90℃for reactionThe reaction was completed by 4h, TLC detection of the starting material, concentration of the reaction solution, purification by column chromatography (V Dichloromethane (dichloromethane) /V Methanol =80/1-30/1), 55mg of fluorescent green solid was obtained in 74% yield. UPLC-MS calculated for C 43 H 41 ClFN 9 O 6 [M+H] + :834.29,found:834.09.UPLC-retention time:5.5min. 1 H NMR(400MHz,DMSO-d 6 )δ11.10(s,1H),10.08(s,1H),8.86(s,2H),7.89(d,J＝8.0Hz,1H),7.80(d,J＝12.0Hz,1H),7.68(s,1H),7.66(s,2H),7.11(d,J＝2.0Hz,1H),6.83(q,J＝4.0Hz,1H),5.11-5.08(m,1H),4.76-4.73(m,2H),4.09(s,6H),3.64-3.61(m,4H),3.09(s,4H),2.93-2.85(m,3H),2.01(s,6H),1.89-1.86(m,2H),1.39(m,2H).
Example 15:
Figure BDA0003259005860000491
raw material 29 (200 mg,1.0 eq.), intermediate 50 (460 mg,1.2 eq.), HATU (650 mg,1.2 eq.) were dissolved in 10mL of anhydrous N, N-dimethylformamide, protected with argon, N-diisopropylethylamine (740 μl,3.0 eq.) was added, and reacted overnight at room temperature, TLC detection, raw material reaction was completed, the reaction solution was added with 20mL of water, extracted with ethyl acetate, and the organic phase was dried over anhydrous sodium sulfate, concentrated, and purified by column chromatography (V Petroleum ether /V Acetic acid ethyl ester =2/1) to give 490mg of white solid in 87% yield. UPLC-MS calculated for C 21 H 15 ClFN 3 O 2 [M+H] + :396.09,found:395.96.UPLC-retention time:6.8min. 1 H NMR(400MHz,DMSO-d 6 )δ10.42(s,1H),8.78(s,1H),8.78-8.43(s,1H),7.88(d,J＝8.0Hz,1H),7.08(s,2H),7.36-7.33(m,1H),7.12(d,J＝4.0Hz,1H),6.83(q,J＝4.0Hz,1H),2.02(s,6H).
Figure BDA0003259005860000492
Intermediate 55 (490 mg,1.0 eq.) raw material 2 (230 mg,1.0 eq.) and potassium carbonate (256 mg,1.5 eq.) were dissolved in 10mL anhydrous N, N-dimethylformamide, argon shield, liter The reaction was carried out overnight at 90℃and TLC showed complete reaction of starting material, concentrated solution, purified by column chromatography (V Petroleum ether /V Acetic acid ethyl ester =4/1) to give 542mg of a white solid, yield 78%. UPLC-MS calculated for C 30 H 32 ClN 5 O 4 [M+H] + :562.22,found:561.90.UPLC-retention time:6.5min. 1 H NMR(400MHz,DMSO-d 6 )δ9.98(s,1H),8.71(s,1H),8.08-8.06(s,1H),7.88(d,J＝8.0Hz,1H),7.68(s,2H),7.12(d,J＝4.0Hz,1H),6.89(d,J＝8.0Hz,1H),6.83(q,J＝4.0Hz,1H),3.62-3.60(m,4H),3.41-3.39(s,1H),2.01(s,6H),1.39(s,9H).
Figure BDA0003259005860000501
Intermediate 56 (500 mg,1.0 eq.) was dissolved in 20mL dichloromethane, 5mL 4m dioxane hydrochloride solution was added, stirred overnight at room temperature, TLC detection, complete reaction of starting material, filtration, dissolution of the solid in 5mL water, pH adjustment to 8-9 with aqueous ammonia, extraction with dichloromethane, drying with anhydrous magnesium sulfate, filtration and concentration to yield 375mg white solid with 91% yield. UPLC-MS calculated for C 25 H 24 ClN 5 O 2 [M+H] + :462.17,found:462.05.UPLC-retention time:3.2min.
Figure BDA0003259005860000502
Intermediate 57 (300 mg,1.0 eq.) and intermediate 5 (165 mg,1.2 eq.) were dissolved in 15mL 1, 2-dichloroethane, 2 drops of acetic acid were added, stirred at room temperature for 0.5h, sodium triacetylborohydride (275 mg,2.0 eq.) was added, reacted overnight at room temperature, TLC detected complete starting material reaction, quenched with 10mL methanol, the reaction concentrated, and purified by column chromatography (V Dichloromethane (dichloromethane) /V Methanol =20/1) to give 380mg of a white solid in 89% yield. UPLC-MS calculated for C 36 H 43 ClN 6 O 4 [M+H] + :659.31,found:659.06.UPLC-retention time:5.1min. 1 H NMR(400MHz,DMSO-d 6 )δ9.96(s,1H),8.03(d,J＝12Hz,2H),7.67(s,2H),7.16(s,1H),6.87-6.80(m,2H),4.45(d,J＝12.0Hz,2H),3.92(s,4H),3.69(s,4H),2.38(s,1H),2.12(d,J＝4.0,2H),2.00(s,6H),1.65(d,J＝8.0,2H),1.57(d,J＝12.0Hz,2H),1.34(s,9H),1.17(s,1H),0.96-0.89(m,2H).
Figure BDA0003259005860000503
Intermediate 58 (350 mg,1.0 eq.) was dissolved in 20mL dichloromethane, 5mL 4m dioxane hydrochloride solution was added, stirred overnight at room temperature, TLC detection, complete reaction of starting material, filtration, dissolution of the solid in 5mL water, pH adjustment to 8-9 with aqueous ammonia, extraction with dichloromethane, drying with anhydrous magnesium sulfate, filtration and concentration to give 280mg white solid with 95% yield. UPLC-MS calculated for C 31 H 35 ClN 6 O 2 [M+H] + :559.25,found:560.02.UPLC-retention time:2.9min.
Figure BDA0003259005860000504
Intermediate 59 (100 mg,1.0 eq.) and intermediate 15 (60 mg,1.2 eq.) were dissolved in 5mL anhydrous N, N-dimethylformamide, protected with argon, N-diisopropylethylamine (62 μl,2.0 eq.) was added, the reaction was allowed to proceed to 90 ℃ for 8h, tlc detected complete reaction of starting materials, the reaction concentrated, and purified by column chromatography (V Dichloromethane (dichloromethane) /V Methanol =80/1-30/1), 99mg of fluorescent green solid was obtained in 68% yield. UPLC-MS calculated for C 44 H 43 ClN 8 O 6 [M+H] + :815.30,found:815.11.UPLC-retention time:6.0min. 1 H NMR(400MHz,DMSO-d 6 )δ10.60(s,1H),9.60(s,1H),8.30(s,1H),7.68(dd,J＝8.0Hz,1.6Hz,1H),7.43(d,J＝8.0Hz,1H),7.19(d,J＝8.0Hz,1H),7.13(s,2H),6.88(s,2H),6.79(d,J＝8.0Hz,1H),6.65(d,J＝2.0Hz,1H),6.58(d,J＝8.0Hz,1H),6.37(dd,J＝4.0Hz,2.4Hz,1H),4.58(dd,J＝8.0Hz,5.6Hz,1H),4.05(d,J＝12.0Hz,2H),3.62(d,J＝16.0Hz,1H),3.15(d,J＝12.0Hz,2H),2.97(s,10H),2.54(t,J＝8.0Hz,2H),2.54-2.48(m,1H),1.35(t,J＝16Hz,2H),0.83-0.73(m,2H).
Example 16:
Figure BDA0003259005860000511
intermediate 90 (50 mg,1.0 eq.) and intermediate 15 (32 mg,1.2 eq.) were dissolved in 5mL anhydrous N, N-dimethylformamide, protected with argon, N-diisopropylethylamine (31 μl,2.0 eq.) was added, the reaction was allowed to proceed to 90 ℃ for 4h, tlc detected complete reaction of starting materials, the reaction concentrated, and purified by column chromatography (V Dichloromethane (dichloromethane) /V Methanol =80/1-30/1), 59mg of fluorescent green solid was obtained in 79% yield. UPLC-MS calculated for C 44 H 42 ClFN 8 O 6 [M+H] + :833.29,found:833.02.UPLC-retention time:6.2min. 1 H NMR(400MHz,DMSO-d 6 )δ11.09(s,1H),10.06(s,1H),8.76(d,J＝4.0Hz,1H),8.15(dd,J＝8.0Hz,2.4Hz,1H),7.89(d,J＝8.0Hz,1H),7.71(d,J＝12.0Hz,1H),7.59(s,2H),7.45(d,J＝4.0Hz,1H),7.12(d,J＝4.0Hz,1H),7.05(d,J＝12.0Hz,1H),7.83(dd,J＝8.0Hz,2.4Hz,1H),5.08(dd,J＝18.0Hz,5.2Hz,1H),4.53(d,J＝16.0Hz,2H),3.62(d,J＝8.0Hz,8H),3.11(s,2H),2.91(t,J＝12.0Hz,2H),2.56(d,J＝16.0Hz,2H),2.07(t,J＝8.0Hz,1H),1.88(d,J＝12.0Hz,2H),1.42-1.34(m,2H).
Example 17:
Figure BDA0003259005860000521
intermediate 55 (400 mg,1.0 eq.) raw material 2 (287 mg,1.0 eq.) and potassium carbonate (210 mg,1.5 eq.) were dissolved in 10mL anhydrous N, N-dimethylformamide under argon, reacted overnight at 90 ℃ with TLC to detect complete reaction of raw material, the reaction solution was concentrated and purified by column chromatography (V Petroleum ether /V Acetic acid ethyl ester =4/1) to give 500mg of a white solid with a yield of 75%. UPLC-MS calculated for C 36 H 43 ClN 6 O 4 [M+H] + :659.31,found:659.03.UPLC-retention time:5.3min. 1 H NMR(400MHz,DMSO-d 6 )δ9.93(s,1H),8.68(s,1H),8.00(dd,J＝8.0Hz,4.0Hz,1H),7.89(d,J＝12.0Hz,1H),7.69(s,2H),7.12(d,J＝2.0Hz,1H),6.86(d,J＝8.0Hz,1H),6.83(q,J＝4.0Hz,1H),4.40(d,J＝12.0Hz,2H),3.27(s,4H),2.86(t,J＝12.0Hz,2H),2.26(s,4H),2.11(d,J＝4.0,2H),2.01(s,6H),1.75(d,J＝8.0Hz,2H),1.36(s,9H),1.19(s,1H),1.03(q,J＝12.0Hz,2H).
Figure BDA0003259005860000522
Intermediate 60 (400 mg,1.0 eq.) was dissolved in 20mL dichloromethane, 5mL 4m dioxane hydrochloride solution was added, stirred overnight at room temperature, TLC detection, complete reaction of the starting material, filtration, dissolution of the solid in 5mL water, pH adjustment to 8-9 with aqueous ammonia, extraction with dichloromethane, drying with anhydrous magnesium sulfate, filtration and concentration to give 320mg white solid with 95% yield. UPLC-MS calculated for C 31 H 35 ClN 6 O 2 [M+H] + :559.25,found:559.09.UPLC-retention time:3.8min.
Figure BDA0003259005860000523
Intermediate 61 (100 mg,1.0 eq.) and intermediate 15 (60 mg,1.2 eq.) were dissolved in 5mL anhydrous N, N-dimethylformamide, protected with argon, N-diisopropylethylamine (62 μl,2.0 eq.) was added, the reaction was warmed to 90 ℃ and allowed to react for 8h, tlc detected complete starting material reaction, the reaction concentrated, and purified by column chromatography (V Dichloromethane (dichloromethane) /V Methanol =80/1-30/1), 92mg of fluorescent green solid was obtained in 63% yield. UPLC-MS calculated for C 44 H 43 ClN 8 O 6 [M+H] + :815.30,found:815.11.UPLC-retention time:5.6min. 1 H NMR(400MHz,DMSO-d 6 )δ11.07(s,1H),9.97(s,1H),8.07(s,2H),8.05(dd,J＝8.0Hz,2.4Hz,1H),7.89(d,J＝8.0Hz,1H),7.75(d,J＝12.0Hz,1H),7.68(s,2H),7.47(s,1H),7.35(dd,J＝8.0Hz,2.0Hz,1H),7.12(d,J＝4.0Hz,1H),6.95(d,J＝12.0Hz,1H),6.83(d,J＝12.0Hz,1H),5.09-5.04(m,1H),4.45(d,J＝12.0Hz,2H),4.19(d,J＝12.0Hz,2H),3.80(s,6H),3.61(s,2H),3.15(s,2H),3.09(s,2H),2.98-2.92(m,1H),2.16(s,1H),2.01(s,6H),1.83(d,J＝12.0Hz,2H),1.22-1.16(m,3H).
Example 18:
Figure BDA0003259005860000531
intermediate 90 (50 mg,1.0 eq.) and intermediate 15 (32 mg,1.2 eq.) were dissolved in 5mL anhydrous N, N-dimethylformamide, protected with argon, N-diisopropylethylamine (31 μl,2.0 eq.) was added, the reaction was allowed to proceed to 90 ℃ for 4h, tlc detected complete reaction of starting materials, the reaction concentrated, and purified by column chromatography (V Dichloromethane (dichloromethane) /V Methanol =80/1-30/1) to yield 57mg of fluorescent green solid in 76%. UPLC-MS calculated for C 44 H 42 ClFN 8 O 6 [M+H] + :833.29,found:833.05.UPLC-retention time:4.7min. 1 H NMR(400MHz,DMSO-d 6 )δ11.09(s,1H),9.99(s,1H),8.69(s,2H),8.05(d,J＝16Hz,1H),7.89(d,J＝8.0Hz,1H),7.80(d,J＝12.0Hz,1H),7.58(s,3H),7.11(s,1H),7.97-6.94(m,1H),6.83(d,J＝12.0Hz,1H),5.12-5.09(m,1H),4.45(d,J＝12.0Hz,2H),4.19(d,J＝12.0Hz,2H),3.89(s,8H),3.15(s,2H),3.09(s,2H),2.98-2.92(m,1H),2.16(s,1H),2.01(s,6H),1.83(d,J＝12.0Hz,2H),1.22-1.16(m,3H).
Example 19:
Figure BDA0003259005860000541
raw material 62 (200 mg,1.0 eq.), intermediate 50 (213 mg,1.2 eq.), HATU (300 mg,1.2 eq.) were dissolved in 10mL anhydrous N, N-dimethylformamide, protected with argon, N-diisopropylethylamine (340 μl,3.0 eq.) was added, the reaction was allowed to proceed overnight at room temperature, TLC was checked, the raw material reaction was complete, the reaction solution was added with 20mL water, extracted with ethyl acetate, the organic phase was dried over anhydrous sodium sulfate, concentrated, and purified by column chromatography (V Petroleum ether /V Acetic acid ethyl ester =2/1) to give 265mg of white solid in 72% yield. UPLC-MS calculated for C 31 H 33 ClFN 4 O 4 [M+H] + :561.22,found:561.02.UPLC-retention time:7.6min.
Figure BDA0003259005860000542
Intermediate 63 (200 mg,1.0 eq.) was dissolved in 20mL dichloromethane, 5mL 4m dioxane hydrochloride solution was added, stirred overnight at room temperature, TLC detection, complete reaction of starting material, filtration, dissolution of the solid in 5mL water, pH adjustment to 8-9 with aqueous ammonia, extraction with dichloromethane, drying with anhydrous magnesium sulfate, filtration and concentration to give 142mg white solid, yield 87%. UPLC-MS calculated for C 26 H 25 ClN 4 O 2 [M+H] + :461.17,found:460.98.UPLC-retention time:2.9min。
Figure BDA0003259005860000543
Intermediate 64 (120 mg,1.0 eq.) and intermediate 5 (67 mg,1.2 eq.) were dissolved in 15mL 1, 2-dichloroethane, 2 drops of acetic acid were added, stirred at room temperature for 0.5h, sodium triacetylborohydride (275 mg,2.0 eq.) was added, reacted overnight at room temperature, TLC detected complete starting material reaction, quenched with 10mL methanol, the reaction concentrated, and purified by column chromatography (V Dichloromethane (dichloromethane) /V Methanol =20/1) to yield 150mg of a white solid in 86% yield. UPLC-MS calculated for C 37 H 44 ClN 5 O 4 [M+H] + :658.31,found:658.13.UPLC-retention time:7.0min. 1 H NMR(400MHz,DMSO-d 6 )δ9.91(s,1H),7.88(d,J＝8.0Hz,2H),7.83(d,J＝8.0Hz,2H),7.60(s,2H),7.12-7.11(m,2H),6.98-6.96(m,1H),6.83(dd,J＝8.0Hz,2.4Hz,1H),3.87(s,6H),2.37(s,4H),2.13(d,J＝4.0Hz,2H),2.01(s,6H),1.79-1.64(m,3H),1.35(s,9H),1.19(s,2H),0.93-0.90(m,2H)
Figure BDA0003259005860000551
Intermediate 65 (100 mg,1.0 eq.) was dissolved in 20mL dichloromethane and 5mL 4m dioxane hydrochloride was addedThe solution was stirred overnight at room temperature, checked by TLC, the starting material reacted completely, filtered, the solid was dissolved in 5mL of water, pH was adjusted to 8-9 with aqueous ammonia, extracted with dichloromethane, dried over anhydrous magnesium sulfate, filtered and concentrated to a white solid 80mg, yield 93%. UPLC-MS calculated for C 32 H 36 ClN 5 O 2 [M+H] + :558.26,found:558.03.UPLC-retention time:3.9min.
Figure BDA0003259005860000552
Intermediate 66 (50 mg,1.0 eq.) and intermediate 15 (30 mg,1.2 eq.) were dissolved in 5mL anhydrous N, N-dimethylformamide, protected with argon, N-diisopropylethylamine (30 μl,2.0 eq.) was added, the reaction was allowed to proceed to 90 ℃ for 8h, tlc detected complete reaction of starting materials, the reaction concentrated, and purified by column chromatography (V Dichloromethane (dichloromethane) /V Methanol =80/1-30/1), 43mg of fluorescent green solid was obtained in 59% yield. UPLC-MS calculated for C 45 H 44 ClN 7 O 6 [M+H] + :814.31,found:814.21.UPLC-retention time:6.7min. 1 H NMR(400MHz,DMSO-d 6 )δ11.05(s,1H),9.98(s,1H),7.90(dd,J＝8.0Hz,2.0Hz,4H),7.65(d,J＝8.0Hz,1H),7.61(s,2H),7.34(s,1H),7.25(d,J＝8.0Hz,1H),7.12(d,J＝4.0Hz,1H),7.09(d,J＝8.0Hz,2H),6.83(dd,J＝8.0Hz,4.0Hz,1H),5.04(dd,J＝8.0Hz,1.2Hz,1H),4.09(d,J＝16.0Hz,2H),4.01(d,J＝16.0Hz,2H),3.61(d,J＝8.0Hz,2H),3.48(s,8H),3.20-3.11(m,3H),2.99(t,J＝8.0Hz,2H),1.83(d,J＝8.0Hz,2H),1.29-1.20(m,2H).
Example 20:
Figure BDA0003259005860000561
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intermediate 66 (30 mg,1.0 eq.) and intermediate 15 (20 mg,1.2 eq.) were dissolved in 5mL anhydrous N, N-dimethylformamide, protected with argon, N-diisopropylethylamine (20 μl,2.0 eq.) was added, the reaction was allowed to proceed to 90 ℃ for 4h, tlc detected complete reaction of starting material, the reaction concentrated, and purified by column chromatography (V Dichloromethane (dichloromethane) /V Methanol =80/1-30/1) to yield 27mg of fluorescent green solid in 61%. UPLC-MS calculated for C 45 H 43 ClFN 7 O 6 [M+H] + :832.29,found:832.17.UPLC-retention time:5.7min. 1 H NMR(400MHz,DMSO-d 6 )δ11.09(s,1H),9.92(s,1H),8.12(s,1H),7.86(dd,J＝18.0,8.8Hz,3H),7.71–7.59(m,3H),7.41(d,J＝7.3Hz,1H),7.15–6.78(m,3H),5.07(dd,J＝12.8,5.3Hz,1H),3.58(d,J＝11.8Hz,4H),2.99–2.51(m,6H),2.33–1.62(m,13H),1.27–1.25(m,2H).
Example 21:
Figure BDA0003259005860000562
Figure BDA0003259005860000571
the raw material 67 (2.0 g) was dissolved in 40mL of methanol, heated to 75 ℃ and refluxed for 6h, TLC detection of the reaction of the raw material was completed, the reaction solution was dried by spin to remove the solvent, ethyl acetate, saturated aqueous sodium bicarbonate extraction, the organic layer was dried, and the solvent was removed by spin evaporation to give 2.0g of a white oily product, yield 95%. UPLC-MS calculated for C 8 H 7 FO 2 [M+H] + :155.04,found:154.85.UPLC-retention time:3.9min. 1 H NMR(400MHz,DMSO-d 6 )δ8.00-7.97(m,2H),7.34-7.29(m,2H),3.81(s,3H).
Figure BDA0003259005860000572
Intermediate 68 (1.0 g,1.0 eq.), intermediate 20 (1.8 g,1.0 eq.), potassium carbonate (1.3 g,1.5 eq.) were dissolved in 10mL anhydrous N, N-dimethylformamide, protected with argon, and the reaction was allowed to warm to 90 ℃ overnight, TLC detected complete starting material reaction, reaction concentrated, column chromatography purified (V Petroleum ether /V Acetic acid ethyl ester =4/1) to give 1.1g of a white solid, yield 39%. UPLC-MS calculated for C 23 H 35 N 3 O 4 [M+H] + :418.26,found:418.08.UPLC-retention time:6.3min.
Figure BDA0003259005860000573
Intermediate 69 (1.0 g,1.0 eq.) was dissolved in 20mL of a mixed solution of tetrahydrofuran and water (V Tetrahydrofuran (THF) /V Water and its preparation method =3/1), heating to 35 ℃ and reacting overnight, TLC detecting that the raw materials are completely reacted, adjusting the pH of the reaction liquid to about 5.0 by using 1M HCl, extracting by using dichloromethane, and concentrating the organic phase by drying to obtain white solid 950mg with the yield of 98%. UPLC-MS calculated for C 22 H 33 N 3 O 4 [M+H] + :405.25,found:405.21.UPLC-retention time:3.5min.
Figure BDA0003259005860000574
Intermediate 70 (500 mg,1.0 eq.), intermediate 50 (400 mg,1.2 eq.), HATU (560 mg,1.2 eq.) were dissolved in 10mL anhydrous N, N-dimethylformamide, protected with argon, N-diisopropylethylamine (650 μl,3.0 eq.) was added, reacted overnight at room temperature, TLC detection, the starting material was complete, the reaction solution was added with 20mL water, extracted with ethyl acetate, the organic phase was dried over anhydrous sodium sulfate, concentrated, and purified by column chromatography (V Petroleum ether /V Acetic acid ethyl ester =3/1) to give 550mg of a white solid, yield 68%. UPLC-MS calculated for C 35 H 42 ClN 7 O 4 [M+H] + :658.31,found:658.21.UPLC-retention time:5.5min. 1 H NMR(400MHz,DMSO-d 6 )δ9.88(s,1H),7.99(d,J＝8.0Hz,1H),7.92-7.87(m,1H),7.82(d,J＝8.0Hz,1H),7.60(d,J＝8.0Hz,2H),7.12(d,J＝4.0Hz,1H),7.07(d,J＝8.0Hz,1H),6.96(d,J＝8.0Hz,1H),6.83(dd,J＝8.0Hz,2.4Hz,1H),4.06-3.84(m,2H),2.85(s,2H),2.69(s,2H),2.26(m,4H),2.13(d,J＝8.0Hz,2H),2.01(s,6H),1.95(s,1H),1.76(d,J＝12.0Hz,2H),1.36(s,9H),1.19(s,2H),1.14(t,J＝4.0Hz,2H).
Figure BDA0003259005860000581
Intermediate 71 (500 mg,1.0 eq.) was dissolved in 60mL dichloromethane, 15mL 4m dioxane hydrochloride solution was added, stirred overnight at room temperature, TLC detection, complete reaction of the starting material, filtration, dissolution of the solid in 30mL water, pH adjustment to 8-9 with aqueous ammonia, extraction with dichloromethane, drying with anhydrous magnesium sulfate, filtration and concentration to give 400mg white solid with 95% yield. UPLC-MS calculated for C 32 H 36 ClN 5 O 2 [M+H] + :558.26,found:558.07.UPLC-retention time:4.2min.
Figure BDA0003259005860000582
Intermediate 72 (100 mg,1.0 eq.) and intermediate 15 (60 mg,1.2 eq.) were dissolved in 5mL anhydrous N, N-dimethylformamide, protected with argon, N-diisopropylethylamine (60 μl,2.0 eq.) was added, the reaction was allowed to proceed to 90 ℃ for 8h, tlc detected complete reaction of starting materials, the reaction concentrated, and purified by column chromatography (V Dichloromethane (dichloromethane) /V Methanol =80/1-30/1), to give 101mg of fluorescent green solid in 69% yield. UPLC-MS calculated for C 45 H 44 ClN 7 O 6 [M+H] + :814.31,found:814.11.UPLC-retention time:5.3min. 1 H NMR(400MHz,DMSO-d 6 )δ11.07(s,1H),9.90(s,1H),7.89(d,J＝8.0Hz,2H),7.85(d,J＝8.0Hz,2H),7.75(d,J＝12.0Hz,1H),7.61(s,2H),7.48(s,1H),7.35(d,J＝12Hz,1H),7.12(s,1H),7.02-6.99(m,1H),6.86-6.82(m,1H),5.07(dd,J＝7.6Hz,5.2Hz,1H),4.19(d,J＝16.0Hz,2H),3.92(d,J＝16.0Hz,1H),3.58(s,8H),3.15-3.09(s,4H),2.84(t,J＝8.0Hz,2H),2.08(s,1H),2.01(s,6H),1.82(d,J＝12.0Hz,2H),1.22-1.16(m,2H).
Example 22:
Figure BDA0003259005860000583
intermediate 72 (50 mg,1.0 eq.) and intermediate 17 (30 mg,1.2 eq.) were dissolved in 5In anhydrous N, N-dimethylformamide (mL), N-diisopropylethylamine (30. Mu.L, 2.0 eq.) was added under argon protection, the reaction was allowed to proceed at 90℃for 4h, the reaction mixture was concentrated, and purified by column chromatography (V Dichloromethane (dichloromethane) /V Methanol =80/1-30/1), 48mg of fluorescent green solid was obtained in 65% yield. UPLC-MS calculated for C 45 H 43 ClFN 7 O 6 [M+H] + :832.30,found:832.16.UPLC-retention time:5.8min. 1 H NMR(400MHz,DMSO-d 6 )δ11.09(s,1H),9.89(s,1H),7.85(dd,J＝24.8,8.8Hz,3H),7.74–7.58(m,3H),7.42(d,J＝7.4Hz,1H),7.12(d,J＝2.3Hz,1H),6.97(d,J＝9.0Hz,2H),6.83(dd,J＝8.7,2.4Hz,1H),5.08(dd,J＝12.8,5.3Hz,1H),3.87(d,J＝12.5Hz,2H),2.93–2.49(m,10H),2.31–1.68(m,15H),1.20–1.16(m,2H).
Conclusion of YMA327 experiments
Figure BDA0003259005860000591
Raw material 73 (600 mg,1.0 eq.) and intermediate 20 (1.2 g,1.2 eq.) were dissolved in 40mL isopropanol, 1 drop of concentrated hydrochloric acid was added, the temperature was raised to 90 ℃ and reacted for 6h, tlc detected complete reaction of the raw material, the reaction solution was concentrated and purified by column chromatography (V Petroleum ether /V Acetic acid ethyl ester =3/1) to give 1.2g of a white solid, yield 80%. UPLC-MS calculated for C 21 H 33 N 5 O 4 [M+H] + :420.03,found:420.26.UPLC-retention time:5.7min. 1 H NMR(400MHz,DMSO-d 6 )δ7.77(d,J＝8.0Hz,1H),7.23(d,J＝8.0Hz,1H),4.48(d,J＝8.0Hz,2H),3.93(s,1H),3.82(s,3H),2.97(t,J＝12.0Hz,1H),2.63(d,J＝8.0Hz,1H),2.29-2.26(m,4H),2.11(d,J＝8.0Hz,1H),1.84-1.76(m,4.0Hz,2H),1.37(s,2H),1.36(s,9H),1.20(m,2H),1.09-1.00(m,2H).
Figure BDA0003259005860000601
Intermediate 74 (1.0 g,1.0 eq.) was dissolved in 20mL of a mixed solution of tetrahydrofuran and water (V Tetrahydrofuran (THF) /V Water and its preparation method =3/1), heating to 35 ℃ and reacting overnight, TLC detecting that the raw materials are completely reacted, adjusting the pH of the reaction liquid to about 5.0 by using 1M HCl, extracting by using dichloromethane, and concentrating the organic phase by drying to obtain white solid 820mg with the yield of 85%. UPLC-MS calculated for C 20 H 31 N 5 O 4 [M+H] + :406.24,found:406.25.UPLC-retention time:2.5min.
Figure BDA0003259005860000602
Intermediate 75 (800 mg,1.0 eq.), intermediate 50 (650 mg,1.2 eq.), HATU (900 mg,1.2 eq.) were dissolved in 10mL anhydrous N, N-dimethylformamide, protected with argon, N-diisopropylethylamine (1000 μl,3.0 eq.) was added, reacted overnight at room temperature, TLC detection, starting material reaction was complete, the reaction solution was added with 20mL water, extracted with ethyl acetate, the organic phase was dried over anhydrous sodium sulfate, concentrated, and purified by column chromatography (V Petroleum ether /V Acetic acid ethyl ester =3/1) to give 1.1g of a white solid in 86% yield. UPLC-MS calculated for C 35 H 42 ClN 7 O 4 [M+H] + :660.30,found:660.27.UPLC-retention time:5.5min.
Figure BDA0003259005860000603
Intermediate 76 (1.0 g,1.0 eq.) was dissolved in 5mL dichloromethane, dioxane hydrochloride 1mL was added, stirred overnight at room temperature, TLC detected complete reaction of the starting material, filtered, the solid dissolved in 10mL water, pH adjusted to 8-9 with aqueous ammonia, extracted with dichloromethane, dried over anhydrous magnesium sulfate, filtered and concentrated to give 790mg of white solid, yield 93%. UPLC-MS calculated for C 30 H 34 ClN 7 O 2 [M+H] + :560.25,found:560.24.UPLC-retention time:4.0min.
Figure BDA0003259005860000604
Intermediate 77 (100 mg,1.0 eq.) and intermediate 15 (60 mg,1.2 eq.) were dissolved in 5mL anhydrous N, N-dimethylformamide, protected with argon, N-diisopropylethylamine (60 μl,2.0 eq.) was added, the reaction was allowed to proceed to 90 ℃ for 8h, tlc detected complete reaction of starting materials, the reaction concentrated, and purified by column chromatography (V Dichloromethane (dichloromethane) /V Methanol =80/1-30/1), to give 82mg of fluorescent green solid in 56% yield. UPLC-MS calculated for C 43 H 42 ClN 9 O 5 [M+H] + :816.30,found:815.96.UPLC-retention time:5.9min. 1 H NMR(400MHz,DMSO-d 6 )δ11.08(s,1H),10.66(s,1H),7.93-7.88(m,2H),7.74(d,J＝4.0Hz,1H),7.46(t,J＝8.0Hz,2H),7.35(d,J＝8.0Hz,1H),7.14-.7.13(m,2H),6.84(q,J＝4.0Hz,2H),5.07(q,J＝8.0Hz,1H),4.53(d,J＝20.0Hz,2H),4.18(d,J＝16.0Hz,2H),3.62(d,J＝16.0Hz,4H),3.36-3.28(m,2H),3.12-3.09(m,4H),3.00-2.94(m,2H),2.89-2.82(m,1H),2.15(s,1H),2.02(s,6H),1.86(d,J＝8.0Hz,2H),1.23(d,J＝12.0Hz,2H),1.19(s,1H).
Example 23:
Figure BDA0003259005860000611
intermediate 77 (50 mg,1.0 eq.) and intermediate 17 (30 mg,1.2 eq.) were dissolved in 5mL anhydrous N, N-dimethylformamide, protected with argon, N-diisopropylethylamine (30 μl,2.0 eq.) was added, heated to 90 ℃ and reacted for 4h, tlc detected complete reaction of starting material, concentrated reaction solution, purified by column chromatography (V Dichloromethane (dichloromethane) /V Methanol =80/1-30/1), 52mg of fluorescent green solid was obtained in 70% yield. UPLC-MS calculated for C 43 H 41 ClFN 9 O 6 [M+H] + :834.29,found:834.14.UPLC-retention time:6.0min. 1 H NMR(400MHz,DMSO-d 6 )δ11.11(s,1H),10.66(s,1H),7.93-7.88(m,2H),7.81(d,J＝8.0Hz,1H),7.76(s,2H),7.69(d,J＝8.0Hz,1H),7.44(d,J＝8.0Hz,1H),7.14-.7.13(m,1H),6.84(q,J＝4.0Hz,1H),5.10(dd,J＝8.0Hz,4.0Hz,1H),4.53(d,J＝8.0Hz,2H),3.78(d,J＝16.0Hz,4H),3.64(m,6H),3.24(m,2H),3.00-2.94(m,2H),2.89-2.82(m,1H),2.15(s,1H),2.02(s,6H),1.87(d,J＝8.0Hz,2H),1.25(d,J＝12.0Hz,2H),1.19(s,1H).
Conclusion of YMA329 experiment
Figure BDA0003259005860000612
Figure BDA0003259005860000621
Raw material 73 (600 mg,1.0 eq.) and raw material 2 (780 mg,1.2 eq.) were dissolved in 40mL isopropanol, 1 drop of concentrated hydrochloric acid was added, the temperature was raised to 90 ℃ and reacted for 6h, tlc detected complete reaction of the raw materials, the reaction solution was concentrated, and purified by column chromatography (V Petroleum ether /V Acetic acid ethyl ester =3/1) to yield 850mg of white solid in 76% yield. UPLC-MS calculated for C 15 H 22 N 4 O 4 [M+H] + :323.17,found:322.96.UPLC-retention time:3.5min. 1 H NMR(400MHz,DMSO-d 6 )δ7.83(d,J＝8.0Hz,1H),7.25(d,J＝4.0Hz,1H),3.84(m,3H),3.73(t,J＝2.4Hz,4H),3.45(t,J＝2.4Hz,4H),1.38(s,9H).
Figure BDA0003259005860000622
Intermediate 74 (800 mg,1.0 eq.) was dissolved in 20mL of a mixed solution of tetrahydrofuran and water (V Tetrahydrofuran (THF) /V Water and its preparation method =3/1), the temperature is raised to 35 ℃ and the reaction is carried out overnight, TLC detects that the raw materials are completely reacted, the pH of the reaction liquid is regulated to about 5.0 by using 1M HCl, the reaction liquid is extracted by using dichloromethane, and the organic phase is dried and concentrated to obtain 790mg of white solid with the yield of 79 percent. UPLC-MS calculated for C 14 H 20 N 4 O 4 [M+H] + :406.24,found:406.25.UPLC-retention time:2.5min.
Figure BDA0003259005860000623
Intermediate 79 (700 mg,1.0 eq.) intermediate 50 (560 mg,1.2 eq.) and HATU @79mg, 1.2 eq.) was dissolved in 10mL anhydrous N, N-dimethylformamide, protected with argon, N-diisopropylethylamine (900 μl,3.0 eq.) was added, reacted overnight at room temperature, TLC detected that the starting material was complete, the reaction solution was added with 20mL water, extracted with ethyl acetate, the organic phase was added to anhydrous sodium sulfate, dried and concentrated, and purified by column chromatography (V Petroleum ether /V Acetic acid ethyl ester =3/1) to give 800mg of white solid in 82% yield. UPLC-MS calculated for C 29 H 31 ClN 6 O 4 [M+H] + :563.21,found:562.98.UPLC-retention time:7.6min.UPLC-MS calculated for C 35 H 42 ClN 7 O 4 [M+H] + :660.30,found:660.29.UPLC-retention time:5.7min. 1 H NMR(400MHz,DMSO-d 6 )δ10.69(s,1H),7.94(d,J＝16.0Hz,1H),7.88(d,J＝16.0Hz,1H),7.74(s,2H),7.39(d,J＝12.0Hz,1H),7.14(d,J＝4.0Hz,1H),6.83(d,J＝4.0Hz,1H),3.73(t,J＝4.0Hz,4H),3.46(s,4H),2.01(s,6H),1.40(s,9H).
Figure BDA0003259005860000631
Intermediate 80 (700 mg,1.0 eq.) was dissolved in 5mL dichloromethane, dioxane hydrochloride 1mL was added, stirred overnight at room temperature, TLC detected complete reaction of the starting material, filtered, the solid dissolved in 10mL water, pH adjusted to 8-9 with aqueous ammonia, extracted with dichloromethane, dried over anhydrous magnesium sulfate, filtered and concentrated to give 510mg of white solid in 89% yield. UPLC-MS calculated for C 24 H 23 ClN 6 O 2 [M+H] + :463.16,found:462.95.UPLC-retention time:5.4min.
Figure BDA0003259005860000632
Intermediate 81 (500 mg,1.0 eq.) and starting material 5 (280 mg,1.2 eq.) were dissolved in 10mL 1, 2-dichloroethane, 1 drop acetic acid was added, and after stirring at room temperature for 0.5h, sodium triacetylborohydride (95 mg,2.0 eq.) was added and reacted overnight at room temperature, TLC detected complete starting material reaction and 5mL methanol was added to quench the reaction The solution was concentrated, purified by column chromatography (V Dichloromethane (dichloromethane) /V Methanol =20/1) to give 584mg of a white solid in 82% yield. UPLC-MS calculated for C 35 H 42 ClN 7 O 4 [M+H] + :660.30,found:660.29.UPLC-retention time:5.7min. 1 H NMR(400MHz,DMSO-d 6 )δ10.53(s,1H),7.89(t,J＝8.0Hz,2H),7.74(s,2H),7.37(d,J＝12.0Hz,1H),7.12(s,1H),6.82(d,J＝8.0Hz,1H),3.90(s,2H),3.70(s,4H),3.12(s,2H),2.42(s,4H),2.01(s,6H),1.69-1.60(m,3H),1.35(s,9H),0.97-0.91(m,4H).
Figure BDA0003259005860000633
Intermediate 82 (500 mg,1.0 eq.) was dissolved in 5mL dichloromethane, added 1mL dioxane hydrochloride, stirred overnight at room temperature, TLC detected complete reaction of the starting material, filtered, the solid dissolved in 10mL water, pH adjusted to 8-9 with aqueous ammonia, extracted with dichloromethane, dried over anhydrous magnesium sulfate, filtered and concentrated to 360mg white solid with 84% yield. UPLC-MS calculated for C 30 H 34 ClN 7 O 2 [M+H] + :560.25,found:560.24.UPLC-retention time:3.2min.
Figure BDA0003259005860000641
Intermediate 83 (100 mg,1.0 eq.) and intermediate 15 (60 mg,1.2 eq.) were dissolved in 5mL anhydrous N, N-dimethylformamide, protected with argon, N-diisopropylethylamine (60 μl,2.0 eq.) was added, the reaction was allowed to proceed to 90 ℃ for 8h, tlc detected complete reaction of starting materials, the reaction concentrated, and purified by column chromatography (V Dichloromethane (dichloromethane) /V Methanol =80/1-30/1), 86mg of fluorescent green solid was obtained in 59% yield. UPLC-MS calculated for C 43 H 42 ClN 9 O 6 [M+H] + :816.30,found:816.15.UPLC-retention time:5.6min. 1 H NMR(400MHz,DMSO-d 6 )δ11.06(s,1H),10.72(s,1H),7.96(dd,J＝53.9,8.9Hz,2H),7.69(d,J＝44.9Hz,4H),7.37–7.12(m,3H),6.86–6.81(m,1H),5.04(d,J＝18.2Hz,1H),4.59(d,J＝9.9Hz,1H),4.08(d,J＝12.8Hz,2H),3.78–3.43(m,4H),3.24–2.54(m,8H),2.02(s,12H),1.31–1.16(m,2H).
Example 24:
Figure BDA0003259005860000642
intermediate 83 (50 mg,1.0 eq.) and intermediate 17 (30 mg,1.2 eq.) were dissolved in 5mL anhydrous N, N-dimethylformamide, protected with argon, N-diisopropylethylamine (30 μl,2.0 eq.) was added, the reaction was allowed to warm to 90 ℃ for 4h, tlc detected complete reaction of starting materials, the reaction concentrated, and purified by column chromatography (V Dichloromethane (dichloromethane) /V Methanol =80/1-30/1) to yield 54mg of fluorescent green solid in 72% yield. UPLC-MS calculated for C 43 H 41 ClFN 9 O 6 [M+H] + :834.29,found:834.14.UPLC-retention time:5.5min. 1 H NMR(400MHz,DMSO-d 6 )δ11.09(s,1H),10.67(s,1H),7.90(t,J＝9.5Hz,2H),7.70(d,J＝31.5Hz,3H),7.40(t,J＝8.5Hz,2H),7.14(s,1H),6.83(d,J＝8.7Hz,1H),5.07(dd,J＝12.8,5.4Hz,1H),3.79–3.55(m,8H),2.92–2.79(m,4H),2.26–1.71(m,15H),1.29–1.26(m,2H).
Example 25:
Figure BDA0003259005860000651
raw material 10 (500 mg,1.0 eq.), raw material 84 (640 mg,1.0 eq.), potassium carbonate (640 mg,1.5 eq.) were dissolved in 10mL anhydrous N, N-dimethylformamide, and reacted overnight at 90℃under argon, and TLC detected that the raw material was completely reacted, the reaction liquid was concentrated, and purified by column chromatography (V) Petroleum ether /V Acetic acid ethyl ester =3/1) to yield 710mg of a white solid in 64% yield. UPLC-MS calculated for C 18 H 17 ClN 2 O 3 [M+H] + :345.10,found:344,96.UPLC-retention time:6.3min. 1 H NMR(400MHz,DMSO-d 6 )δ7.93(d,J＝12.0Hz,1H),7.47(d,J＝8.0Hz,2H),7.31(d,J＝4.0Hz,1H),7.26(d,J＝8.0Hz,1H),7.03(q,J＝4.0Hz,1H),1.35(s,9H).
Figure BDA0003259005860000652
Intermediate 85 (710 mg,1.0 eq.) was dissolved in 20mL dichloromethane, 5mL 4m dioxane hydrochloride solution was added, stirred overnight at room temperature, the starting material was complete by TLC, filtered, the solid was dissolved in 5mL water, pH was adjusted to 8-9 with aqueous ammonia, extracted with dichloromethane, dried over anhydrous magnesium sulfate, filtered and concentrated to a white solid 485mg, 96% yield. UPLC-MS calculated for C 13 H 9 ClN 2 O[M+H] + :245.04,found:244.92.UPLC-retention time:3.1min. 1 H NMR(400MHz,DMSO-d 6 )δ7.93(d,J＝12.0Hz,1H),7.47(d,J＝8.0Hz,2H),7.31(d,J＝4.0Hz,1H),7.26(d,J＝8.0Hz,1H),7.03(q,J＝4.0Hz,1H),5.71(s,2H).
Figure BDA0003259005860000661
Intermediate 86 (481mg, 1.2 eq.), starting 62 (505 mg,1.0 eq.), HATU (753 mg,1.2 eq.) were dissolved in 10mL anhydrous N, N-dimethylformamide, protected with argon, N-diisopropylethylamine (863 μl,3.0 eq.) was added, reacted overnight at room temperature, TLC detected, starting was complete, the reaction mixture was added with 20mL water, extracted with ethyl acetate, the organic phase was dried over anhydrous sodium sulfate, concentrated, and purified by column chromatography (V Petroleum ether /V Acetic acid ethyl ester =3/1) to yield 712mg of a white solid in 81%. UPLC-MS calculated for C 29 H 29 ClN 4 O 4 [M+H] + :533.19,found:533.31.UPLC-retention time:6.6min. 1 H NMR(400MHz,DMSO-d 6 )δ10.07(s,1H),7.91(d,J＝8.0Hz,1H),7.85(d,J＝12.0Hz,1H),7.24(s,1H),7.16-7.13(m,2H),7.00(d,J＝12.0Hz,3H),3.43(s,4H),3.26(s,4H),1.39(s,9H).
Figure BDA0003259005860000662
Intermediate 87 (710 mg,1.0 eq.) was dissolved in 20mL dichloromethane, 5mL 4m dioxane hydrochloride solution was added, stirred overnight at room temperature, the starting material was complete by TLC, filtered, the solid was dissolved in 10mL water, pH was adjusted to 8-9 with aqueous ammonia, extracted with dichloromethane, dried over anhydrous magnesium sulfate, filtered and concentrated to a white solid 554mg, 96% yield. UPLC-MS calculated for C 24 H 21 ClN 4 O 2 [M+H] + :432.90,found:432.95.UPLC-retention time:3.1min.
Figure BDA0003259005860000663
Intermediate 88 (554 mg,1.0 eq.) and intermediate 5 (327 mg,1.2 eq.) were dissolved in 15mL 1, 2-dichloroethane, 2 drops of acetic acid were added, stirred at room temperature for 0.5h, sodium triacetylborohydride (552 mg,2.0 eq.) was added, reacted overnight at room temperature, TLC detected complete starting material reaction, quenched with 10mL methanol, concentrated, purified by column chromatography (V Dichloromethane (dichloromethane) /V Methanol =20/1) to yield 660mg of a white solid in 82% yield. UPLC-MS calculated for C 35 H 40 ClN 5 O 4 [M+H] + :630.28,found:630.15.UPLC-retention time:5.9min.
Figure BDA0003259005860000664
Intermediate 89 (660 mg,1.0 eq.) was dissolved in 20mL dichloromethane, 5mL 4m dioxane hcl solution was added, stirred overnight at room temperature, the starting material was complete by TLC, filtered, the solid was dissolved in 10mL water, pH adjusted to 8-9 with aqueous ammonia, extracted with dichloromethane, dried over anhydrous magnesium sulfate, filtered and concentrated to give 511mg white solid with 92% yield. UPLC-MS calculated for C 24 H 21 ClN 4 O 2 [M+H] + :530.23,found:529.92.UPLC-retention time:4.0min.
Figure BDA0003259005860000671
Intermediate 90 (50 mg,1.0 eq.) and intermediate 15 (31 mg,1.2 eq.) were dissolved in 5mL anhydrous N, N-dimethylformamide, protected with argon, N-diisopropylethylamine (33 μl,2.0 eq.) was added, the reaction was warmed to 90 ℃ and allowed to react for 8h, tlc detected complete starting material reaction, the reaction concentrated, and purified by column chromatography (V Dichloromethane (dichloromethane) /V Methanol =80/1-30/1), 46mg of fluorescent green solid was obtained in 62% yield. UPLC-MS calculated for C 43 H 40 ClN 7 O 6 [M+H] + :786.27found:786.00.UPLC-retention time:5.8min. 1 H NMR(400MHz,DMSO-d 6 )δ11.06(s,1H),10.06(s,1H),7.92(d,J＝12.0Hz,1H),7.86(d,J＝8.0Hz,4H),7.62(d,J＝8.0Hz,1H),7.29(s,1H),7.25(d,J＝8.0Hz,1H),7.21(d,J＝8.0Hz,1H),7.15(d,J＝12.0Hz,3H),7.02(d,J＝2.0Hz,1H),6.99(d,J＝2.0Hz,1H),5.03(dd,J＝8.0Hz,4.0Hz,1H),4.03(d,J＝16.0Hz,6H),3.36-3.29(m,3H),2.95(t,J＝12.0Hz,2H),2.63-2.59(m,4H),2.93(s,2H),1.99-1.96(m,2H),1.81(d,J＝12.0Hz,2H),1.20-1.13(m,2H).
Example 26:
Figure BDA0003259005860000672
raw material 91 (500 mg,1.0 eq.), raw material 84 (460 mg,1.0 eq.), potassium carbonate (685 mg,1.5 eq.) were dissolved in 10mL anhydrous N, N-dimethylformamide, and reacted overnight at 90℃under argon, and TLC detection of the completion of the raw material reaction, concentration of the reaction solution, purification by column chromatography (V) Petroleum ether /V Acetic acid ethyl ester =3/1) to yield 788mg of white solid in 70%. UPLC-MS calculated for C 19 H 20 N 2 O 4 [M+H] + :341.15,found:340.94.UPLC-retention time:5.7min. 1 H NMR(400MHz,DMSO-d 6 )δ7.96(d,J＝12.0Hz,1H),7.47(d,J＝8.0Hz,2H),7.31(d,J＝4.0Hz,1H),7.26(d,J＝8.0Hz,1H),7.03(q,J＝4.0Hz,1H),3.83(s,9H),1.35(s,9H).
Figure BDA0003259005860000681
Intermediate 92 (788 mg,1.0 eq.) was dissolved in 20mL dichloromethane, 5mL 4m dioxane hydrochloride solution was added, stirred overnight at room temperature, the starting material was complete by TLC, filtered, the solid was dissolved in 5mL water, pH was adjusted to 8-9 with aqueous ammonia, extracted with dichloromethane, dried over anhydrous magnesium sulfate, filtered and concentrated to give 534mg white solid with 96% yield. UPLC-MS calculated for C 12 H 12 N 2 O 2 [M+H] + :241.09,found:241.13.UPLC-retention time:3.0min. 1 H NMR(400MHz,DMSO-d 6 )δ7.96(d,J＝12.0Hz,1H),7.47(d,J＝8.0Hz,2H),7.31(d,J＝4.0Hz,1H),7.26(d,J＝8.0Hz,1H),7.03(q,J＝4.0Hz,1H),4.78(s,9H),3.83(s,9H).
Figure BDA0003259005860000682
Intermediate 93 (378 mg,1.2 eq.) raw 62 (400 mg,1.0 eq.) HATU (596 mg,1.2 eq.) was dissolved in 10mL anhydrous N, N-dimethylformamide, protected with argon, N-diisopropylethylamine (810 μl,3.0 eq.) was added, reacted overnight at room temperature, TLC detected that the raw materials were completely reacted, the reaction mixture was added with 20mL water, extracted with ethyl acetate, the organic phase was dried over anhydrous sodium sulfate, concentrated, and purified by column chromatography (V Petroleum ether /V Acetic acid ethyl ester =2/1) to yield 580mg of white solid in 84% yield. UPLC-MS calculated for C 30 H 32 N 4 O 5 [M+H] + :529.24,found:592.31.UPLC-retention time:6.2min.
Figure BDA0003259005860000683
Intermediate 94 (580 mg,1.0 eq.) was dissolved in 20mL of dichloromethane, 5mL of 4M dioxane hydrochloride solution was added, stirring was continued overnight at room temperature, TLC was checked, the starting material was complete, filtration was performed, the solid was dissolved in 5mL of water, pH was adjusted to 8-9 with ammonia, dichloromethane extraction was performed, anhydrous magnesium sulfate was added, drying was performed, filtration was performed, and concentration was performed451mg of white solid was obtained in 96% yield. UPLC-MS calculated for C 25 H 24 N 4 O 3 [M+H] + :429.18,found:428.98.UPLC-retention time:3.8min.
Figure BDA0003259005860000691
Intermediate 95 (457mg, 1.0 eq.) and intermediate 5 (268 mg,1.2 eq.) were dissolved in 15mL of 1, 2-dichloroethane, 2 drops of acetic acid were added, stirred at room temperature for 0.5h, sodium triacetylborohydride (4476 mg,2.0 eq.) was added, reacted overnight at room temperature, TLC detected complete starting material reaction, quenched with 10mL of methanol, concentrated, and purified by column chromatography (V Dichloromethane (dichloromethane) /V Methanol =20/1) to give 552mg of a white solid in 84% yield. UPLC-MS calculated for C 36 H 43 N 5 O 5 [M+H] + :626.33,found:626.28.UPLC-retention time:5.9min.
Figure BDA0003259005860000692
Intermediate 95 (552 mg,1.0 eq.) was dissolved in 20mL dichloromethane, 5mL 4m dioxane hydrochloride solution was added, stirred overnight at room temperature, the starting material was detected by TLC, the reaction was complete, filtered, the solid was dissolved in 5mL water, pH was adjusted to 8-9 with aqueous ammonia, extracted with dichloromethane, dried over anhydrous magnesium sulfate, filtered and concentrated to a white solid 445mg, 96% yield. UPLC-MS calculated for C 31 H 35 N 5 O 3 [M+H] + :526.27,found:526.28.UPLC-retention time:3.8min.
Figure BDA0003259005860000693
Intermediate 97 (50 mg,1.0 eq.) and intermediate 15 (31 mg,1.2 eq.) were dissolved in 5mL anhydrous N, N-dimethylformamide under argon, N-diisopropylethylamine (33. Mu.L, 2.0 eq.) was added and reacted for 8h at 90℃with TLC detection of starting material reactionThe reaction solution was concentrated and purified by column chromatography (V Dichloromethane (dichloromethane) /V Methanol =80/1-30/1), 48mg of fluorescent green solid was obtained in 65% yield. UPLC-MS calculated for C 44 H 43 N 7 O 7 [M+H] + :782.32,found:782.00.UPLC-retention time:5.9min. 1 H NMR(400MHz,DMSO-d 6 )δ11.07(s,1H),10.12(s,1H),7.91(d,J＝8.0Hz,2H),7.84(d,J＝8.0Hz,2H),7.65(d,J＝8.0Hz,2H),7.33(s,1H),7.24(d,J＝8.0Hz,1H),7.13-7.07(m,4H),6.83(d,J＝4.0Hz,1H),6.46(dd,J＝8.0Hz,2.0Hz,1H),6.46(dd,J＝8.0Hz,2.0Hz,1H),5.04(dd,J＝8.0Hz,5.4Hz,1H),4.07(d,J＝8.0Hz,2H),3.99(d,J＝8.0Hz,2H),3.87(s,3H),3.66-3.64(m,4H),3.25(t,J＝12.0Hz,2H),3.14-3.08(m,4H),2.98(t,J＝12.0Hz,2H),2.85(s,1H),2.17(s,1H),1.99-1.96(m,1H),1.87(d,J＝12.0Hz,2H),1.30-1.19(m,2H).
Example 27:
Figure BDA0003259005860000701
raw material 8 (240 mg,1.2 eq.), raw material 62 (500 mg,1.0 eq.), HATU (750 mg,1.2 eq.) were dissolved in 10mL of anhydrous N, N-dimethylformamide, protected with argon, N-diisopropylethylamine (860 μl,3.0 eq.) was added, the reaction was allowed to proceed overnight at room temperature, TLC detection, the raw material reaction was completed, the reaction solution was added with 20mL of water, extracted with ethyl acetate, the organic phase was dried over anhydrous sodium sulfate, concentrated, and purified by column chromatography (V Petroleum ether /V Acetic acid ethyl ester =3/1) to give 500mg of a white solid with a yield of 75%. UPLC-MS calculated for C 23 H 29 N 3 O 4 [M+H] + :412.22,found:412.01.UPLC-retention time:4.8min.
Figure BDA0003259005860000702
Intermediate 98 (500 mg,1.0 eq.) raw material 10 (190 mg,1.0 eq.) and potassium carbonate (250 mg,1.5 eq.) were dissolved in 10mL anhydrous N, N-dimethylformamide under argon, reacted overnight at 90℃under elevated temperature, and the reaction was detected by TLCTo completion, the reaction solution was concentrated, and purified by column chromatography (V Petroleum ether /V Acetic acid ethyl ester =3/1) to give 553mg of a white solid in 83% yield. UPLC-MS calculated for C 30 H 21 ClN 4 O 4 [M+H] + :547.21,found:547.02.UPLC-retention time:7.2min.
Figure BDA0003259005860000703
Intermediate 99 (553 mg,1.0 eq.) was dissolved in 20mL dichloromethane, 5mL 4m dioxane hydrochloride solution was added, stirred overnight at room temperature, the starting material was detected by TLC, complete reaction, filtered, the solid was dissolved in 5mL water, pH was adjusted to 8-9 with aqueous ammonia, extracted with dichloromethane, dried over anhydrous magnesium sulfate, filtered and concentrated to a white solid 434mg, 96% yield. UPLC-MS calculated for C 25 H 23 ClN 4 O 2 [M+H] + :448.15,found:448.02.UPLC-retention time:3.8min.
Figure BDA0003259005860000711
Intermediate 100 (400 mg,1.0 eq.) and starting material 5 (230 mg,1.2 eq.) were dissolved in 15mL 1, 2-dichloroethane, 2 drops of acetic acid were added, stirred at room temperature for 0.5h, sodium triacetylborohydride (380 mg,2.0 eq.) was added, reacted overnight at room temperature, TLC detected complete starting material reaction, 10mL methanol quench reaction was added, the reaction solution was concentrated, and column chromatography was performed to purify (V Dichloromethane (dichloromethane) /V Methanol =20/1) to yield 420mg of a white solid in 73% yield. UPLC-MS calculated for C 36 H 42 ClN 5 O 4 [M+H] + :644.30found:644.00.UPLC-retention time:5.3min.
Figure BDA0003259005860000712
Intermediate 101 (420 mg,1.0 eq.) was dissolved in 20mL dichloromethane and 5mL 4m dioxane hydrochloride solution was added at room temperatureStirring overnight, TLC detection, complete reaction of raw materials, filtering, dissolving the solid in 5mL of water, adjusting pH to 8-9 with ammonia water, extracting with dichloromethane, adding anhydrous magnesium sulfate for drying, filtering, concentrating to obtain white solid 337mg, and yield 95%. UPLC-MS calculated for C 31 H 34 ClN 5 O 2 [M+H] + :543.97,found:544.24.UPLC-retention time:2.6min.
Figure BDA0003259005860000713
Intermediate 102 (100 mg,1.0 eq.) and intermediate 15 (61 mg,1.2 eq.) were dissolved in 5mL anhydrous N, N-dimethylformamide, protected with argon, N-diisopropylethylamine (65 μl,2.0 eq.) was added, the reaction was allowed to proceed to 90 ℃ for 8h, tlc detected complete reaction of starting materials, the reaction concentrated, and purified by column chromatography (V Dichloromethane (dichloromethane) /V Methanol =80/1-30/1), 89mg of fluorescent green solid was obtained in 61% yield. UPLC-MS calculated for C 44 H 43 ClN 7 O 6 [M+H] + :800.29,found:800.04.UPLC-retention time:5.9min. 1 H NMR(400MHz,DMSO-d 6 )δ11.06(s,1H),10.12(s,1H),7.90(d,J＝8.0Hz,2H),7.80(d,J＝8.0Hz,2H),7.70(d,J＝12.0Hz,1H),7.63(d,J＝12.0Hz,2H),7.31(s,1H),7.23(d,J＝12.0Hz,1H),7.16(s,1H),7.07(d,J＝4.0Hz,2H),6.92-6.89(m,1H),5.04(dd,J＝8.0Hz,5.4Hz,1H),4.05(d,J＝8.0Hz,2H),3.93(s,2H),3.66-3.64(m,4H),3.25(t,J＝12.0Hz,2H),3.14-3.08(m,4H),2.97(t,J＝12.0Hz,2H),2.63-2.57(m,2H),2.06(s,3H),1.99-1.95(m,1H),1.90(d,J＝4.0Hz,2H),1.24-1.19(m,2H).
Example 28:
Figure BDA0003259005860000721
/>
intermediate 102 (50 mg,1.0 eq.) and intermediate 17 (32 mg,1.2 eq.) were dissolved in 5mL anhydrous N, N-dimethylformamide, protected with argon, N-diisopropylethylamine (33. Mu.L, 2.0 eq.) was added, the reaction was allowed to proceed to 90℃for 4h, TLC detection of starting material reaction was complete, the reaction solution was concentrated, and column chromatography was pure Formation (V) Dichloromethane (dichloromethane) /V Methanol =80/1-30/1), 60mg of fluorescent green solid was obtained in 80% yield. UPLC-MS calculated for C 44 H 41 ClFN 7 O 6 [M+H] + :818.28,found:818.14.UPLC-retention time:5.3min. 1 H NMR(400MHz,DMSO-d 6 )δ11.09(s,1H),10.06(s,1H),7.91(d,J＝8.0Hz,3H),7.79(s,1H),7.72-7.68(m,2H),7.45(d,J＝8.0Hz,1H),7.16(s,1H),7.11-7.06(m,3H),6.91(d,J＝8.0Hz,1H),5.08(dd,J＝8.0Hz,4Hz,1H),3.99(s,2H),3.61(s,6H),3.16-3.13(m,6H),2.96-2.83(m,3H),2.07(s,3H),2.01(s,2H),1.88(d,J＝8.0Hz,2H),1.43-1.35(m,2H).
< biological evaluation test >
Example 29: evaluation of Androgen Receptor (AR) expression level reduction effect
AR positive human prostate cancer cells LNCaP, 22RV1 and VCaP were respectively treated in RPMI 1640 or DMEM (hereinafter referred to as evaluation medium) containing 10% FBS to reach 3X 10 5 Amount per well was inoculated into 6-well microwell plates (Corning) and incubated overnight. To this culture, an evaluation medium containing the compound of the example was added so that the final concentration of the compound of the example reached 0.1 and 1. Mu. Mol/L, and the culture was continued for 24 hours. Then removing the culture medium, washing cells by using PBS, adding a RIPA lysate containing 1%Protease Inhibitor Cocktail, and obtaining a total protein extract after lysis and centrifugation, wherein the protein concentration in the extract is detected by a BCA method; protein electrophoresis was performed by SDS-PAGE, after which 200mA constant current was transferred for 90min to transfer the protein to PVDF (MilliporeSigma IPVH 00010) membrane; placing the PVDF film in skimmed milk containing 5%, and sealing for 1h at room temperature; using Anti-Androgen Receptor antibody [ EPR1535 (2)](HRP) (abcam) for immune response; and (3) dripping ECL luminous liquid after washing the film, and exposing. The strips were greyscale analyzed using software Image J. Each sample was tested simultaneously for GAPDH, alpha-tublin or beta-Actin protein bands as internal controls. The degradation rate of the compound of the example on the AR protein is calculated according to the protein band gray scale, and is shown in the figures 1-3
The results of LNCaP, VCaP and 22RV1 cells are shown in Table 2 below. The decrease in AR expression was 50% or more and was indicated as "decrease". Among the compounds of the present invention, the example compounds YMA-301, YMA-302, YMA-303, YMA-304, YMA-305, YMA-306, YMA-307, YMA-308, TYA-309, TYA-310, YMA-315, YMA-316, TYA-317, TYA-318, YMA-323, YMA-324 and YMA-325 exhibited a certain AR expression reducing effect on three prostate cancer cells of LNCaP, VCaP and 22RV 1.
Reduction of Androgen Receptor (AR) expression level by Compounds of Table 2
Figure BDA0003259005860000731
EXAMPLE 30 androgen-dependent prostate cancer cell proliferation inhibitory Activity
Androgen receptor positive human prostate cancer cells LNCaP were cultured in RPMI 1640 medium containing 5% carbon-adsorbed serum (CCS) (hereinafter referred to as evaluation medium) to achieve 5X 10 3 The amount of/well was inoculated into a transparent bottom 96-well microplate (Corning) and cultured for 48 hours. To this culture were added an evaluation medium containing R1881 (final concentration of R1881: 0.1nmo 1/L) and an evaluation medium containing the compound of example or comparative example (Enzalutamide: enzalutamide). (the final concentrations of the compounds of this example or comparative example were 3.810395, 11.43, 34.29, 102.88, 308.64, 925.9, 2777.78, 8333.33 and 25000nmo 1/L), and after 96 hours of culture, the number of living cells was determined. Viable cell count was determined using WST-1 (Roche). The proliferation activity value of R1881 at 0.1nmol/L was set to 100%, the proliferation activity of the cells in the evaluation medium alone was set to 0%, and the 50% proliferation Inhibition Concentration (IC) was calculated from the number of living cells measured by logistic regression 50 Values).
The results of cell proliferation inhibition are shown in FIG. 4, which shows that compound YMA-325 exhibits androgen-dependent prostate cancer cell proliferation inhibition activity, with a maximum half inhibition concentration (IC 50 ) 106.9nmol/L. Enzalutamide as positive control, IC 50 36.6nmo1/L.
The foregoing description is only of the preferred embodiments of the present application and is not intended to limit the same, but rather, various modifications and variations may be made by those skilled in the art. Any modification, equivalent replacement, improvement, etc. made within the spirit and principles of the present application should be included in the protection scope of the present application.

Claims (6)

1. A compound represented by the general formula (I) or a pharmaceutically acceptable salt, stereoisomer thereof,
Figure QLYQS_1
general formula (I)
Wherein Q is Linker and A is E3 ubiquitin ligase moiety;
r1, R3, R4 are independently selected from H, halogen, -C 1－8 An alkyl group;
r2 is selected from O;
the ring of R5-R8 is phenyl,
q is a divalent linking group, and has the following structure:
Figure QLYQS_2
* Junction of left and right sides of Q
A is a targeted E3 ubiquitin ligase ligand, and the structure is as follows:
Figure QLYQS_3
wherein R9 is independently selected from hydrogen.
2. The compound represented by the general formula (I) or a pharmaceutically acceptable salt, stereoisomer thereof according to claim 1, wherein the compound represented by the general formula (I) is selected from:
Figure QLYQS_4
3. A pharmaceutical composition comprising a compound of general formula (I) according to any one of claims 1-2, or a pharmaceutically acceptable salt, stereoisomer thereof, and at least one pharmaceutically acceptable carrier or excipient.
4. The pharmaceutical composition according to claim 3, wherein the pharmaceutical dosage form is an oral preparation or an injection preparation.
5. Use of a compound of general formula (I) according to any one of claims 1-2 or a pharmaceutically acceptable salt, stereoisomer thereof, or a pharmaceutical composition according to claim 3 for the manufacture of a medicament for an anti-androgenic agent.
6. Use of a compound of general formula (I) according to any one of claims 1-2, or a pharmaceutically acceptable salt, stereoisomer thereof, for the manufacture of a medicament for a disease or disorder associated with Androgen Receptor (AR) expression, said disease or disorder being cancer, metastatic bone disease, prostatic hyperplasia, acne vulgaris, seborrhea, hirsutism, androgenic alopecia, precocious puberty or andropathy.
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CN110506039A (en) * 2016-10-11 2019-11-26 阿尔维纳斯股份有限公司 Compounds and methods for for androgen receptor targeting degradation
CN111825657A (en) * 2019-04-18 2020-10-27 成都海创药业有限公司 Dual-function chimeric heterocyclic compound for targeted degradation of androgen receptor and application thereof
WO2021081108A1 (en) * 2019-10-22 2021-04-29 Arvinas Operations, Inc. Methods of treating prostate cancer

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* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN110506039A (en) * 2016-10-11 2019-11-26 阿尔维纳斯股份有限公司 Compounds and methods for for androgen receptor targeting degradation
CN111825657A (en) * 2019-04-18 2020-10-27 成都海创药业有限公司 Dual-function chimeric heterocyclic compound for targeted degradation of androgen receptor and application thereof
WO2021081108A1 (en) * 2019-10-22 2021-04-29 Arvinas Operations, Inc. Methods of treating prostate cancer

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