CN115960094A - Preparation and application of aromatic ring substituted methylamine derivative - Google Patents
Preparation and application of aromatic ring substituted methylamine derivative Download PDFInfo
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- CN115960094A CN115960094A CN202211077205.0A CN202211077205A CN115960094A CN 115960094 A CN115960094 A CN 115960094A CN 202211077205 A CN202211077205 A CN 202211077205A CN 115960094 A CN115960094 A CN 115960094A
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Abstract
The invention discloses a preparation method and application of aromatic ring substituted methylamine derivatives. The aromatic ring substituted methylamine derivative has a structure shown in a formula I, and is expected to be used for treating tumor patients carrying p 53Y 220C mutation.
Description
Technical Field
The invention mainly relates to preparation and application of aromatic ring substituted methylamine derivatives.
Background
The TP53 gene, also known as P53, encodes a protein with a molecular weight of 53kDa, P53 is an anti-cancer gene, which is mutated in more than 50% of all malignancies, and the protein encoded by this gene is a transcription factor that controls the initiation of the cell cycle and plays a crucial role from the beginning of cell division (biomoles 2020,10,303 doi. In 1979 scientists discovered the P53 protein. The P53 protein was first recognized by scientists as a cellular chaperone for the large T antigen of monkey kidney virus (SV) 40 and as an oncoprotein of SV virus (naturereviews | CanCer FeBRUARy 2009, volumeme 9). Over 10 years of research shows that the P53 protein is not oncoprotein but oncostatin, and if the cell is damaged and cannot be repaired, the P53 protein participates in the initiation process, so that the cell dies in apoptosis. The human P53 gene is a very important cancer suppressor gene, which is expressed in normal cells to a low degree and in malignant tumors to a high degree. The cancer suppressor gene 'P53' in the cell can judge the degree of DNA variation, if the variation is small, the gene can promote cell self-repair, and if the DNA variation is large, the 'P53' can induce cell apoptosis. P53 is actually a transcription factor that is induced to be expressed when cells are under stress, thereby promoting the cells to enter the arrest phase of the cell cycle, and thus regulating the apoptotic or senescence signaling pathway. The P53 protein translated from the P53 gene is an important regulator of cell growth, proliferation and injury repair. When the DNA of the cell is damaged, the P53 protein prevents the cell from stopping in the G1/S phase, repairs the damage, and promotes the apoptosis if the damage cannot be repaired. Cells with P53 mutations continue to divide under adverse conditions to form tumor cells. Mutations of P53 in tumor cells are very common, and the main goal today is to develop anti-cancer drugs based on P53 muteins.
At present, small molecules aiming at P53 are mainly PPI (protein-protein interaction) inhibitors, such as MDM2-P53 inhibitors, and at present, stable activators directly acting on P53 mutant protein can benefit patients with P53 specific mutation, and the mutation of P53-Y220 accounts for 1.0-1.5% of all tumors, so that no drug is approved at present.
Disclosure of Invention
The invention aims to solve the technical problem that no effective medicine is approved for a cancer patient with P53 mutation at present, so that the invention provides preparation and application of aromatic ring substituted methylamine derivatives. The compound is expected to be used for treating tumor patients carrying p 53Y 220C mutation.
The present invention provides a compound represented by formula I, or an isotopic derivative thereof, or a pharmaceutically acceptable salt of any one of the foregoing (i.e., the compound represented by formula I or the isotopic derivative), or a solvate of any one of the foregoing (i.e., the compound represented by formula I, the isotopic derivative, or the pharmaceutically acceptable salt):
wherein ring D is
(# and # are used to identify the connection location);
ring D 1 And ring D 2 In each A 1 、A 2 、A 3 And A 4 Each independently is N or CH;
ring D 3 In (B) 1 、B 3 And B 4 Each independently is CH, N, NH, O or S; b is 2 And B 5 Each independently is C or N;
each R a Independently F, cl, br, I, -CN, C 1 -C 4 Alkyl radical, C 1 -C 4 Alkoxy or halo C 1 -C 4 An alkyl group;
m is
Z 1 Is C (R) 3 ) N, O or N (R) 4 )；
Z 2 Is C (R) 5 ) N, O or N (R) 6 )；
Z 3 Is CH or N;
Z 4 is CH or N;
each L 2 Independently is-NH-, -CH 2 -, -O-or-S-;
ring Q 1 And ring Q 2 Each independently is a saturated or partially unsaturated 5-to 10-membered carbocyclic ring, a saturated or partially unsaturated 5-to 10-membered heterocyclic ring, a 6-to 10-membered aromatic ring, a 5-to 10-membered heteroaromatic ring, or an 8-to 10-membered bicyclic fused ring; one ring in the 8-10 membered bicyclic fused ring is a saturated or partially unsaturated 5-6 membered carbocyclic ring or a saturated or partially unsaturated 5-6 membered heterocyclic ring, and the other ring is a benzene ring or a 5-6 membered heteroaromatic ring;
ring D 3 And in ring C, the- -bond is a double bond or a single bond, provided that ring D 3 And ring C is an aromatic ring;
R 3 、R 4 、R 5 and R 6 Each independently is H, C 1 -C 4 Alkyl or halo C 1 -C 4 An alkyl group;
R A and R B Independently H, halogen or C 1 -C 4 An alkyl group;
L 1 is absent or-C (O) -;
ring B is a saturated or partially unsaturated 3-to 10-membered carbocyclic ring, a saturated or partially unsaturated 3-to 10-membered heterocyclic ring, a 6-to 10-membered aromatic ring, a 5-to 10-membered heteroaromatic ring, or a 6-to 10-membered bicyclic fused ring; one ring in the 6-10 membered bicyclic fused ring is a saturated or partially unsaturated 5-6 membered carbocyclic ring or a saturated or partially unsaturated 5-6 membered heterocyclic ring, and the other ring is a benzene ring or a 5-6 membered heteroaromatic ring;
each R 2 Independently F, cl, br, I, = O, -CN, -NH 2 、-NHR d 、-NR d R e 、C 1 -C 4 Alkyl, by 1,2 or 3R c Substituted C 1 -C 4 Alkyl radical, C 1 -C 4 Alkoxy, by 1,2 or 3R c Substituted C 1 -C 4 Alkoxy, halo C 1 -C 4 Alkyl, -C (O) NHR d 、-C(O)NR d R e 、-C(O)OR d or-SO 2 R d ；
Each R 1 Independently F, cl, br, I, -NH 2 、＝O、-NHR d 、-NR d R e 、C 1 -C 4 Alkyl, by 1,2 or 3R h Substituted C 1 -C 4 Alkyl radical, C 1 -C 4 Alkoxy, by 1,2 or 3R h Substituted C 1 -C 4 Alkoxy, halo C 1 -C 4 Alkyl, 3-to 10-membered cycloalkyl or 3-to 10-membered heterocycloalkyl (e.g.
)；
Each R d And R e Each independently is H, C 1 -C 4 Alkyl or by 1,2 or 3R b Substituted C 1 -C 4 An alkyl group;
each R c 、R b And R h Each independently is-OH, -OR f 、-NH 2 、-NHR f or-NR f R g ；
Each R f And R g Each independently is C 1 -C 4 Alkyl or-Boc;
n is 0,1, 2,3 or 4;
m is 0,1, 2,3 or 4;
p is 0,1, 2,3 or 4;
the number of heteroatoms in the heterocycle, heterocycloalkyl, heteroaryl ring and heteroaryl group is 1,2 or 3, and each heteroatom is independently N, O or S.
In some embodiments, in a compound of formula I, an isotopic derivative thereof, or a pharmaceutically acceptable salt of any of the foregoing, or a solvate of any of the foregoing (i.e., a compound of formula I, an isotopic derivative, or a pharmaceutically acceptable salt of the foregoing), ring D is
(# and # are used to identify the connection location);
ring D 1 And ring D 2 In each A 1 、A 2 、A 3 And A 4 Each independently is N or CH;
ring D 3 In (B) 1 、B 3 And B 4 Each independently is CH, N, NH, O or S; b 2 And B 5 Each independently is C or N;
each R a Independently F or-CN;
m is
Z 1 Is N (R) 4 )；
Z 2 Is C (R) 5 )；
Z 3 Is N;
Z 4 is CH;
each L 2 Independently of one anotheris-NH-;
ring Q 1 And ring Q 2 Each independently is a saturated or partially unsaturated 5-to 10-membered carbocyclic ring, a saturated or partially unsaturated 5-to 10-membered heterocyclic ring, a 6-to 10-membered aromatic ring, a 5-to 10-membered heteroaromatic ring, or an 8-to 10-membered bicyclic fused ring; one ring in the 8-10 membered bicyclic fused ring is a saturated or partially unsaturated 5-6 membered carbocyclic ring or a saturated or partially unsaturated 5-6 membered heterocyclic ring, and the other ring is a benzene ring or a 5-6 membered heteroaromatic ring;
ring D 3 And in ring C, the- -bond is a double bond or a single bond, provided that ring D 3 And ring C is an aromatic ring;
R 4 and R 5 Each independently of the other is H or halogeno-C 1 -C 4 An alkyl group;
R A is H or C 1 -C 4 An alkyl group;
R B is H or halogen;
L 1 is absent or-C (O) -;
ring B is a saturated or partially unsaturated 3-to 10-membered carbocyclic ring, a saturated or partially unsaturated 3-to 10-membered heterocyclic ring, a 6-to 10-membered aromatic ring, a 5-to 10-membered heteroaromatic ring, or a 6-to 10-membered bicyclic fused ring; one ring in the 6-10 membered bicyclic fused ring is a saturated or partially unsaturated 5-6 membered carbocyclic ring or a saturated or partially unsaturated 5-6 membered heterocyclic ring, and the other ring is a benzene ring or a 5-6 membered heteroaromatic ring;
each R 2 Independently F, = O, -CN, -NH 2 、-NHR d 、C 1 -C 4 Alkyl, by 1,2 or 3R c Substituted C 1 -C 4 Alkyl radical, C 1 -C 4 Alkoxy, halo C 1 -C 4 Alkyl, -C (O) NHR d or-SO 2 R d ；
Each R 1 Independently F, = O, -NR d R e 、C 1 -C 4 Alkyl, by 1,2 or 3R h Substituted C 1 -C 4 Alkyl or 3-10 membered heterocycloalkyl;
each R d And R e Each independently is H or C 1 -C 4 An alkyl group;
each one of whichR c 、R b And R h Each independently is-OH, -OR f 、-NH 2 、-NHR f or-NR f R g ；
Each R f And R g Each independently is C 1 -C 4 Alkyl or-Boc;
n is 0,1 or 2;
m is 0,1 or 2;
p is 0 or 1;
the number of heteroatoms in the heterocycle, heterocycloalkyl, heteroaryl ring and heteroaryl group is 1,2 or 3, and each heteroatom is independently N, O or S.
In some embodiments, the structure of formula I is as shown in formula II:
wherein ring D is
Ring D 1 And ring D 2 In each A 1 、A 2 、A 3 And A 4 Each independently is N or CH;
ring D 3 In (B) 1 、B 3 And B 4 Each independently is CH, N, NH, O or S; b is 2 And B 5 Each independently is C or N;
each R a Independently is F, cl, br, I, C 1 -C 4 Alkyl radical, C 1 -C 4 Alkoxy or halo C 1 -C 4 An alkyl group;
m is
Z 1 Is C (R) 3 ) N, O or N (R) 4 )；
Z 2 Is C (R) 5 ) N, O or N (R) 6 )；
Z 3 Is CH or N;
Z 4 is CH or N;
each L 2 Independently is-NH-, -CH 2 -, -O-or-S-;
ring Q 1 And ring Q 2 Each independently is a saturated or partially unsaturated 5-10 membered carbocyclic ring, a saturated or partially unsaturated 5-10 membered heterocyclic ring, a 6-10 membered aromatic ring, a 5-10 membered heteroaromatic ring, or an 8-10 membered bicyclic fused ring; one ring in the 8-10 membered bicyclic fused ring is a saturated or partially unsaturated 5-6 membered carbocyclic ring or a saturated or partially unsaturated 5-6 membered heterocyclic ring, and the other ring is a benzene ring or a 5-6 membered heteroaromatic ring;
ring D 3 And in ring C, the- -bond is a double bond or a single bond, provided that ring D 3 And ring C is an aromatic ring;
R 3 、R 4 、R 5 and R 6 Each independently is H, C 1 -C 4 Alkyl or halo C 1 -C 4 An alkyl group;
L 1 is absent or-C (O) -;
ring B is a saturated or partially unsaturated 3-to 10-membered carbocyclic ring, a saturated or partially unsaturated 3-to 10-membered heterocyclic ring, a 6-to 10-membered aromatic ring, a 5-to 10-membered heteroaromatic ring, or a 6-to 10-membered bicyclic fused ring; one ring in the 6-10 membered bicyclic fused ring is a saturated or partially unsaturated 5-6 membered carbocyclic ring or a saturated or partially unsaturated 5-6 membered heterocyclic ring, and the other ring is a benzene ring or a 5-6 membered heteroaromatic ring;
each R 2 Independently F, cl, br, I, -NH 2 、-NHR d 、-NR d R e 、C 1 -C 4 Alkyl, by 1,2 or 3R c Substituted C 1 -C 4 Alkyl radical, C 1 -C 4 Alkoxy, with 1,2 or 3R c Substituted C 1 -C 4 Alkoxy, halo C 1 -C 4 Alkyl, -C (O) NHR d 、-C(O)NR d R e 、-C(O)OR d or-SO 2 R d ；
Each R 1 Independently F, cl, br, I, -NH 2 、＝O、-NHR d 、-NR d R e 、C 1 -C 4 Alkyl, by 1,2 or 3R h Substituted C 1 -C 4 Alkyl radical, C 1 -C 4 Alkoxy, by 1,2 or 3R h Substituted C 1 -C 4 Alkoxy, halo C 1 -C 4 Alkyl, 3-10 membered cycloalkyl or 3-10 membered heterocycloalkyl;
each R d And R e Each independently is C 1 -C 4 Alkyl or by 1,2 or 3R b Substituted C 1 -C 4 An alkyl group;
each R c 、R b And R h Each independently is-OH, -OR f 、-NH 2 、-NHR f or-NR f R g ；
Each R f And R g Each independently is C 1 -C 4 An alkyl group;
n is 0,1, 2,3 or 4;
m is 0,1, 2,3 or 4;
p is 0,1, 2,3 or 4;
the number of heteroatoms in the heterocycle, heterocycloalkyl, heteroaryl ring and heteroaryl group is 1,2 or 3, and each heteroatom is independently N, O or S.
In some embodiments, when ring Q 1 And ring Q 2 When each is independently a saturated or partially unsaturated 5-to 10-membered carbocyclic ring, the saturated 5-to 10-membered carbocyclic ring is a 5-to 6-membered carbocyclic ring, for example, cyclohexane; the unsaturated 5-10 membered carbocycle is a benzene ring.
In some embodiments, when ring Q 1 And ring Q 2 When each is independently a saturated or partially unsaturated 5-to 10-membered heterocyclic ring, said saturated 5-to 10-membered heterocyclic ring is
In some embodiments, when ring Q 1 And ring Q 2 When each is independently a 6-to 10-membered aromatic ring, the 6-to 10-membered aromatic ring is a benzene ring.
In some embodiments, when ring Q 1 And ring Q 2 When each is independently an 8-10 membered bicyclic fused ring, one of the 8-10 membered bicyclic fused rings is a partially unsaturated 5-6 membered heterocyclic ring and the other ring is a benzene ring, for example,
in some embodiments, ring D 1 Is composed of
In some embodiments, ring D 2 Is composed of
In some embodiments, ring D 3 Is composed of
In some embodiments, ring D is
In some embodiments of the present invention, the substrate is,
is composed of
In some embodiments, Z 4 Is CH.
In some embodiments, Z 1 Is C (R) 3 ) Or N (R) 4 )。
In some embodiments, Z 2 Is C (R) 5 ) N or N (R) 6 )。
In some embodiments of the present invention, the substrate is,
has the structure of
For example
In some embodiments, M is
In some embodiments, Z 3 Is N.
In some embodiments, M is
In some embodiments, M is
And ring D is->
In some embodiments, L 1 Is absent.
In some embodiments, L is 1 is-C (O) -.
In some embodiments, R 3 Is halo C 1 -C 4 Alkyl radicals, e.g. fluoro C 1 -C 4 Alkyl radicals, e.g. fluoro C 2 Alkyl radicals, e.g. -CH 2 CF 3 。
In some embodiments, R 4 Is halo C 1 -C 4 Alkyl radicals, e.g. fluoro C 1 -C 4 Alkyl radicals, e.g. fluoro C 2 Alkyl radicals, e.g. -CH 2 CF 3 。
In some embodiments, R 5 Is H.
In some embodiments, R 6 Is H.
In some embodiments, p is 0 or 1.
In some embodiments, R a Is F or-CN.
In some embodiments of the present invention, the substrate is,
has the structure of
In some embodiments of the present invention, the substrate is,
is structurally characterized as->
In some embodiments of the present invention, the substrate is,
is->
In some embodiments of the present invention, the substrate is,
is->
In some embodiments of the present invention, the substrate is,
is->
In some embodiments, each L is 2 Independently is-NH-.
In some embodiments, ring Q 1 And Q 2 Each independently is
In some embodiments, ring Q 1 And Q 2 Each independently is
(/>
Is represented by L 2 The location of the connection).
In some embodiments, ring Q 1 And Q 2 Each independently is
In some embodiments, each R is 1 Independently F, = O, CH 3 、
In some embodiments of the present invention, the substrate is,
are each independently->
m is 0,1 or 2 i Is R 1 ，R j Is H or R 1 ，R 1 As defined herein.
In some embodiments of the present invention, the substrate is,
are each independently->
m is 0,1 or 2 i Is R 1 ，R j Is H or R 1 ，R 1 As defined herein.
In some embodiments of the present invention, the substrate is,
can be structured as->
In some embodiments, R i Is F, cl, br, I or C 1 -C 4 Alkyl, such as F or methyl.
In some embodiments, R j Is H, -NH 2 、-NHR d 、-NR d R e 、C 1 -C 4 Alkyl, by 1,2 or 3R h Substituted C 1 -C 4 Alkyl radical, C 1 -C 4 Alkoxy, by 1,2 or 3R h Substituted C 1 -C 4 Alkoxy, halo C 1 -C 4 Alkyl, 3-10 membered cycloalkyl or 3-10 membered heterocycloalkyl.
In some embodiments, each R is d And R e Each independently is C 1 -C 4 Alkyl groups, such as methyl.
In some embodiments, ring Q 1 And ring Q 2 Carbon atom of (1) with L 2 And (4) connecting.
In some embodiments of the present invention, the substrate is,
are each independently>
/>
In some embodiments of the present invention, the substrate is,
can be structured as->
(i.e. trans structure) or
(i.e., cis-structure).
In some embodiments of the present invention, the substrate is,
can be structured as>
In some embodiments of the present invention, the substrate is,
are each independently->
In some embodimentsAnd ring B is
(e.g.. Based on:>
)、
(e.g., based on a predetermined condition>
)
(e.g.. Sup. Ml)>
) Or->
(e.g.. Sup. Ml)>
) E.g. based on>
In some embodiments of the present invention, the substrate is,
is composed of
/>
In some embodiments, each R is independently selected from R, and R 2 Independently F, -NH 2 、-CH 3 、-NHCH 3 、-SO 2 CH 3 、-OCH 3 、-CF 3 、-C(O)NH 2 、＝O、-CN、-CH 2 NHBoc、-CH 2 NH 2 or-C (O) NHCH 3 。
In some embodiments, n is 0,1, or 2.
In some embodiments of the present invention, the substrate is,
is->
In some embodiments of the present invention, the substrate is,
is->
/>
In some embodiments of the present invention, the substrate is,
is composed of
In some embodiments, the compound of formula I has any one of the following structures:
the variables are defined as described herein.
In some embodiments, the compound has any one of the following structures:
in some embodiments, the compound is any one of the following stereoisomers:
the invention also provides a preparation method of the compound shown in the formula I, which can be a method 1 or a method 2:
the method 1 comprises the following steps: in a solvent (for example, a mixed solvent of N, N-dimethylformamide and water), a compound represented by the formula II and a compound represented by the formula III are reacted in the presence of a palladium catalyst (for example, pd (PPh) 3 ) 4 ) And a base (e.g., potassium carbonate) to provide a compound of formula I; wherein X is halogen (e.g. I), [ B ]]Is boric acid (e.g.
) Or a borate ester (e.g.. Based on->
) (ii) a M is->
The definition of each variable is as described above;
the method 2 comprises the following steps: removing Boc from the compound shown in the formula IV in the presence of acid to obtain a compound shown in the formula I; in the compounds of the formula I M is
The definition of each variable is as described above;
the preparation method of the compound shown in the formula IV can comprise the following steps: in a solvent (e.g., a mixed solvent of acetic acid and 1, 2-dichloroethane), a compound represented by the formula IV-1 is mixed with
Carrying out reductive amination reaction in the presence of a reducing agent (such as sodium triacetoxyborohydride) to obtain a compound shown as a formula IV; wherein, L in the compound shown as the formula IV 2 is-NH-, ring Q 2 Carbon atom of (1) with L 2 Connecting; the definition of each variable is as described above;
the preparation method of the compound represented by the formula IV-1 may comprise the following steps: reducing the nitro group of the compound shown in the formula IV-2 (such as Fe powder reduction) to obtain a compound shown in the formula IV-1; the definition of each variable is as described above;
the preparation method of the compound represented by the formula IV-2 may comprise the steps of: carrying out the following coupling reaction of the compound represented by the formula IV-3 and the compound represented by the formula IV-4 in a solvent (such as N, N-dimethylformamide) in the presence of a catalyst (such as cuprous iodide) and a base (such as potassium carbonate) to obtain a compound represented by the formula IV-2; wherein Hal is halogen (e.g. Br or I), and Z in the compound of formula IV-2 3 Is N; the definition of each variable is as described above;
the invention also provides compounds of formula II, III, IV-1, IV-2 or IV-4:
wherein the definitions of the variables are as described above.
The invention also provides a pharmaceutical composition, which comprises the compound, the isotope derivative, the pharmaceutically acceptable salt or the solvate, and at least one pharmaceutic adjuvant.
The invention also provides application of the compound, the isotope derivative, the pharmaceutically acceptable salt or the solvate or the pharmaceutical composition in preparing medicines for treating tumor patients carrying p 53Y 220C mutation.
The present invention also provides a method of treating a patient having a tumor bearing a p 53Y 220C mutation comprising administering to the patient a therapeutically effective amount of a compound, isotopic derivative, pharmaceutically acceptable salt or solvate as described above.
In some embodiments, the tumor may be gastric cancer or liver cancer.
Definitions and explanations
The following terms and phrases used herein are intended to have the following meanings unless otherwise indicated. A particular term or phrase, unless otherwise specifically defined, should not be considered as indefinite or unclear, but rather construed according to ordinary meaning. When a trade name appears herein, it is intended to refer to its corresponding commodity or its active ingredient.
In the present invention, the term "substituted" or "substituent" is that a hydrogen atom in a group is replaced by the specified group. When the position of substitution is not indicated, the substitution may be at any position, but formation of a stable or chemically feasible chemical is permissible. Examples are as follows:
the structure represents that the hydrogen atom on the ring A is substituted by m R 1 And (4) substitution.
When any variable (e.g., R) occurs more than one time in the composition or structure of a compound, its definition in each case is independent. Thus, for example, if a group is substituted with 0-2R, the group may optionally be substituted with up to two R, and there are separate options for R in each case. Furthermore, combinations of substituents and/or variants thereof are permissible only if such combinations result in stable compounds.
When the number of one linking group is absent, it means that the linking group is a single bond, for example, in A-L-Z, L is absent, the structure formed is A-Z.
In the present invention, the term "alkyl" refers to a saturated straight or branched chain monovalent hydrocarbon group. C 1 -C 4 Alkyl means an alkyl group having 1 to 4 carbon atoms, which is specifically methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl or tert-butyl.
In the present invention, the term "haloalkyl" refers to a group formed by substituting one or more (e.g., 2,3,4, 5, or 6) hydrogen atoms in an alkyl group with a halogen, wherein each halogen is independently F, cl, br, or I. Halogen substituted C 1 -C 4 Alkyl means C substituted by one or more halogens 1 -C 4 Alkyl radical, wherein C 1 -C 4 The alkyl group is as defined above. In some embodiments, halo C 1 -C 4 Alkyl being fluoro C 1 -C 4 An alkyl group.
In the present invention, the term "alkoxy" refers to an-O-alkyl group, wherein alkyl is as previously defined. C 1 -C 4 Alkoxy means-O- (C) 1 -C 4 Alkyl) in which C 1 -C 4 The alkyl group is as defined above.
In the present invention, the term "carbocyclic ring" refers to a saturated, partially unsaturated, or aromatic monocyclic or polycyclic (e.g., fused, spiro, or bridged) cyclic group formed by carbon atoms. In a saturated carbocycle, each carbon atom in the ring is saturated, and examples of saturated carbocycles include, but are not limited to
In the aromatic carbocyclic ring, each ring is aromatic, and examples of the aromatic carbocyclic ring include, but are not limited to
In a partially unsaturated carbocyclic ring, at least one carbon atom in the ring is saturated and at least one carbon atom in the ring is unsaturated, examples of partially unsaturated carbocycles include, but are not limited to:>
the 3-to 10-membered carbocyclic ring may specifically be a 3-, 4-, 5-, 6-, 7-, 8-, 9-or 10-membered carbocyclic ring. The 5-to 10-membered carbocyclic ring may specifically be a 5-, 6-, 7-, 8-, 9-or 10-membered carbocyclic ring.
In the present invention, the term "heterocycle" refers to a saturated, partially unsaturated, or aromatic monocyclic or polycyclic (e.g., bicyclic, spirocyclic, or bridged) cyclic group formed by a carbon atom and at least one heteroatom independently selected from N, O, and S. In saturated heterocycles, both the carbon and the heteroatom of the ring are saturated, examples of saturated heterocycles include, but are not limited to
In the aromatic heterocyclic ring, each ring is aromatic, and examples of the aromatic heterocyclic ring include, but are not limited to
Examples of partially unsaturated heterocycles, in which at least one atom in the ring is saturated and at least one atom in the ring is unsaturated, include, but are not limited to>
The 3-to 10-membered heterocyclic ring may specifically be 3,4, 5,6.7, 8, 9 or 10 membered heterocyclic ring. The 5-to 10-membered heterocyclic ring may specifically be a 5-, 6-, 7-, 8-, 9-or 10-membered heterocyclic ring.
In the present invention, the term "aromatic ring" refers to an aromatic carbocyclic ring, each of which is aromatic. The 6-to 10-membered aromatic ring may specifically be a benzene ring or a naphthalene ring.
In the present invention, the term "heteroaromatic ring" refers to an aromatic heterocyclic ring, wherein each ring is aromatic. Examples of heteroaromatic rings include, but are not limited to
The 5-10 membered heteroaromatic ring may specifically be a 5,6,7,8, 9 or 10 membered heteroaromatic ring.
In the present invention, the term "bicyclic fused ring" means a fused ring composed of two single rings, and the linking site thereof to other structures may be located on either single ring. The 6-10 membered bicyclic fused ring may specifically be a 6,7,8, 9 or 10 membered bicyclic fused ring. The 8-10 membered bicyclic fused ring may specifically be an 8, 9 or 10 membered bicyclic fused ring.
In the present invention, the term "cycloalkyl" refers to a monocyclic or polycyclic (e.g., fused, spiro, or bridged) monovalent hydrocarbon group in which each carbon atom is saturated. The 3-to 10-membered cycloalkyl group may specifically be a 3-, 4-, 5-, 6-, 7-, 8-, 9-or 10-membered cycloalkyl group, including cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl and the like. Specific examples of cycloalkyl groups include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl,
In the present invention, the term "heterocycloalkyl group" refers to a group formed by replacing at least one carbon atom in a cycloalkyl group with a heteroatom selected from N, O and S. The 3-10 membered heterocycloalkyl group may specifically be a 3,4, 5,6,7,8, 9 or 10 membered heterocycloalkyl group. Heterocyclic alkyl radicalsExamples include, but are not limited to
The compounds of the present invention and their structures are also meant to include all isomeric (including stereoisomers and tautomers, where stereoisomers such as enantiomers, diastereomers, geometric isomers (e.g., cis-trans isomers) and conformational isomers) forms. They can be defined as (R) -/(S) -or (D) -/(L) -or (R, R) -/(R, S) -/(S, S) -, according to the absolute stereochemistry for amino acids. The present invention includes all of these possible isomers, as well as their racemic, enantiomerically enriched, and optionally pure forms. Optically active (+) and (-), (R) -and (S) -and (R, R) -/(R, S) -/(S, S) -or (D) -and (L) -isomers can be prepared using chiral starting material synthesis, chiral resolution, or can be resolved using conventional techniques such as, but not limited to, high Performance Liquid Chromatography (HPLC) using a chiral column. When the compounds described herein contain an alkenyl double bond or other geometrically asymmetric center, the compounds include both E and Z geometric isomers unless otherwise specified. In the chemical structure, a bond
No configuration is specified, i.e. if there is configurational isomerism in the chemical structure, the bond->
Can be->
Or->
Or at the same time comprise
And &>
Two configurations. Likewise, all tautomeric forms are also included.
In the present invention, the term "tautomer" refers to a proton that moves from one atom of a molecule to another in the same molecule from the original position. The invention includes tautomers of any of the compounds.
In the present invention, the term "isotopic derivative" refers to a compound which differs in structure only in the presence of one or more isotopically enriched atoms. For example, having the structure of the invention except that "deuterium" or "tritium" is used in place of hydrogen, or 18 F-fluorine labeling: ( 18 Isotope of F) instead of fluorine, or with 11 C-， 13 C-, or 14 C-enriched carbon (C) 11 C-， 13 C-, or 14 C-carbon labeling; 11 C-， 13 c-, or 14 C-isotopes) instead of carbon atoms are within the scope of the invention. Such compounds are useful as analytical tools or probes in, for example, biological assays, or may be used as tracers for in vivo diagnostic imaging of disease, or as tracers for pharmacodynamic, pharmacokinetic or receptor studies. In the present invention, the isotopic derivative is, for example, a deuteron.
In the present invention, the term "pharmaceutically acceptable" refers to a substance (e.g., carrier or diluent) that does not affect the biological activity or properties of the compounds of the present invention and is relatively non-toxic, i.e., the substance can be administered to an individual without causing an adverse biological response or interacting in an adverse manner with any of the components contained in the composition.
In the present invention, the term "pharmaceutically acceptable salt" means a salt formed from a suitable non-toxic organic acid, inorganic acid, organic base or inorganic base and a compound, which retains the biological activity of the compound. The organic acid may be any of various organic acids capable of forming a salt, which are conventional in the art, and is preferably one or more of methanesulfonic acid, p-toluenesulfonic acid, maleic acid, fumaric acid, citric acid, tartaric acid, malic acid, lactic acid, formic acid, acetic acid, propionic acid, trifluoroacetic acid, oxalic acid, succinic acid, benzoic acid, isethionic acid, naphthalenesulfonic acid, and salicylic acid. The inorganic acid may be any of various inorganic acids capable of forming a salt, which are conventional in the art, and preferably one or more of hydrochloric acid, sulfuric acid and phosphoric acid. The organic base can be various organic bases which are conventional in the field and can form salts, and one or more of pyridine, imidazole, pyrazine, indole, purine, tertiary amine and aniline is/are preferable. The tertiary amine organic base is preferably triethylamine and/or N, N-diisopropylethylamine. The aniline organic base is preferably N, N-dimethylaniline. The pyridine organic base is preferably one or more of pyridine, picoline, 4-dimethylamino pyridine and 2-methyl-5-ethyl pyridine. The inorganic base may be any of various inorganic bases capable of forming a salt, which are conventional in the art, and preferably one or more of alkali metal hydride, alkali metal hydroxide, alkali metal alkoxide, potassium carbonate, sodium carbonate, lithium carbonate, cesium carbonate, potassium hydrogen carbonate and sodium hydrogen carbonate. The alkali metal hydride is preferably sodium hydride and/or potassium hydride. The alkali metal hydroxide is preferably one or more of sodium hydroxide, potassium hydroxide and lithium hydroxide. The alkoxide of the alkali metal is preferably one or more of sodium methoxide, sodium ethoxide, potassium tert-butoxide and sodium tert-butoxide.
In the present invention, the term "solvate" means a substance formed by a compound or a salt thereof and a suitable solvent. The solvent is preferably water or an organic solvent.
In the present invention, the term "patient" includes any animal, preferably a mammal, more preferably a human.
The above preferred conditions can be arbitrarily combined to obtain preferred embodiments of the present invention without departing from the common general knowledge in the art.
The reagents and starting materials used in the present invention are commercially available.
The positive progress effects of the invention are as follows: the compound of the invention has inhibitory activity on NUGC3 cells and is expected to be used for treating patients with tumors carrying p 53Y 220C mutations.
Detailed Description
The invention is further illustrated by the following examples, which are not intended to limit the scope of the invention. The experimental methods without specifying specific conditions in the following examples were selected according to the conventional methods and conditions, or according to the commercial instructions.
Abbreviations referred to in the examples and their full names are as follows:
HOBT 1-hydroxybenzotriazole
EDCI 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride
DMF N, N-dimethylformamide
Pd(PPh 3 ) 4 Tetrakis (triphenylphosphine) palladium
Pd 2 (dba) 3 Tris (dibenzylideneacetone) dipalladium
Pd(dppf)Cl 2 1,1' -bis-diphenylphosphino ferrocene palladium dichloride
EDTA ethylene diamine tetraacetic acid
HATU 2- (7-azabenzotriazole) -N, N, N ', N' -tetramethyluronium hexafluorophosphate
DIEA N, N-diisopropylethylamine
S-Phos 2-dicyclohexylphosphine-2 ',6' -dimethoxybiphenyl
LDA lithium diisopropylamide
In the following examples, when the purification was carried out using an automatic column chromatography (Biotage), the stationary phase used was a silica gel column (40-63. Mu.M).
Synthesis of intermediate A
Step 1: synthesis of Compound A-1
To the reaction flask were added 3-methoxy-4-nitrobenzoic acid (10.00g, 50.72mmol), dichloromethane (100 mL), methylamine hydrochloride (6.85g, 101.45mmol), HOBT (10.28g, 76.09mmol), EDCI (14.59g, 76.09mmol), triethylamine (20.53g, 202.90mmol). The mixture was stirred at room temperature overnight. After the reaction is finished, the reaction solution is diluted by dichloromethane and water. Separation of organic substancesPhase, aqueous phase dichloromethane extraction. The organic phases were combined, washed with saturated aqueous sodium chloride solution, dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and the crude product obtained was purified by recrystallization from anhydrous ethanol to give the title compound A-1 (6.50g, 61%) as a pale yellowish brown solid. LC-MS (ESI) M/z 211.1 (M + H) + .
And 2, step: synthesis of Compound A
To a reaction flask was added A-1 (6.50g, 30.92mmol), ethanol (60 mL), saturated NH 4 Aqueous Cl solution (20 mL), iron powder (8.63g, 154.62mmol). After the mixture was stirred at 90 ℃ for 1 hour, the reaction solution was concentrated under reduced pressure to remove most of the ethanol. Concentrate was diluted with ethyl acetate and water. The organic phase was separated and the aqueous phase was extracted with ethyl acetate. The organic phases were combined, washed with saturated aqueous sodium chloride solution, dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to give the title compound a as a pale yellow solid (4.70g, 84%). LC-MS (ESI) M/z 181.2 (M + H) + 。 1 H NMR (400 MHz, deuterated chloroform) δ 7.34 (d, J =1.9hz, 1h), 7.13 (dd, J =8.1,1.9hz, 1h), 6.62 (d, J =8.1hz, 1h), 6.29 (s, 1H), 4.09 (s, 2H), 3.84 (s, 3H), 2.95 (d, J =4.8hz, 3h).
Synthesis of intermediate B
Step 1: synthesis of Compound B-1
To the flask was added 2-iodo-4-nitro-1H-indole (1.00g, 3.47mmol), anhydrous tetrahydrofuran (20 mL). The mixture was cooled to 0 ℃ under nitrogen and sodium hydride (690mg, 17.36mmol) was added. After the addition, the temperature is raised to room temperature and the mixture is stirred for 0.5 hour, CF is added 3 CH 2 OTf (3.22g, 13.89mmol). After the reaction mixture was stirred at room temperature for 2 hours, the reaction mixture was cooled to 0 ℃. Slowly adding water to quench the reaction, and then adding ethyl acetate and water to dilute. The organic phase was separated and the aqueous phase was extracted with ethyl acetate. The organic phases were combined, washed with saturated aqueous sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to give the title compound B-1 (1.50 g,>99％)。LC-MS(ESI):m/z371.0(M+H) + .
step 2: synthesis of Compound B-2
To the reaction flask were added B-1 (1.28g, 3.46mmol), acetic acid (10 mL), and iron powder (1.16g, 20.75mmol). The mixture was stirred at 70 ℃ for 4 hours and then cooled to room temperature. Add ethyl acetate and water to dilute. The organic phase was separated and the aqueous phase was extracted with ethyl acetate. The organic phases were combined, washed with saturated aqueous sodium chloride solution, concentrated under reduced pressure, and the crude product was purified by an automatic column chromatography (Biotage) (mobile phase: petroleum ether/ethyl acetate 100/0 to 80/20) to give the target compound B-2 (820 mg, 69%) as a reddish brown solid. LC-MS (ESI) M/z 341.0 (M + H) + .
And 3, step 3: compound B
A reaction flask was charged with B-2 (820mg, 2.41mmol), anhydrous ethanol (30 mL), 4-dimethylaminocyclohexanone (1.7g, 12.06mmol), and ethyl titanate (2.75g, 12.06mmol). The mixture was stirred at 50 ℃ for 5 hours under nitrogen and then cooled to 0 ℃. Sodium cyanoborohydride (760mg, 12.06mmol) was added, and the mixture was stirred at 0 ℃ for 5 minutes, then, it was raised to 50 ℃ and stirred for 1 hour. After the reaction, the reaction solution was directly concentrated under reduced pressure, and the crude product was purified by column chromatography (Biotage) (mobile phase: dichloromethane/methanol 100/0 to 88/12) to give a white solid as intermediate B, LC-MS (ESI): M/z 466.09 (M + H) + .
And (3) synthesis of an intermediate C:
step 1: synthesis of Compound C
To a reaction flask was added 2-methoxy-4-methylsulfonyl-aniline (189mg, 0.94mmol), (3-bromomethylphenyl) boronic acid pinacol ester (866mg, 2.91mmol), tetrahydrofuran (5 mL), cesium carbonate (987 mg, 3.00mmol), and the mixture was heated to reflux and stirred overnight. The next day, the reaction solution was cooled to room temperature and diluted with ethyl acetate and water. The organic phase was separated and the aqueous phase was extracted with ethyl acetate. The combined organic phases were washed with saturated aqueous sodium chloride, dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure and purified by an automatic column chromatography (Biotage) (mobile phase: petroleum ether/ethyl acetate 100/0 to 76/24) to give compound C as a white solid (57mg, 55%). 1 H NMR (400 MHz, deuterated chloroform) δ 7.86-7.71 (m, 3H), 7.54-7.28 (m, 3H), 7.29-7.16 (m, 1H), 6.59 (d, J =8.4hz, 1h), 4.55 (s, 0.8H), 4.40 (s, 2H), 3.91 (s, 3H), 3.01 (s, 3H), 1.35 (d, J =2.7hz, 12h).
Synthesis of intermediate D:
step 1: synthesis of Compound D-1
4-Nitroindole (10.00g, 61.67mmol), anhydrous tetrahydrofuran (100 mL) was added to the reaction flask. The mixture was cooled to 0 ℃ under nitrogen and NaH (7.40g, 185.02mmol) was added in three portions. After the addition, the mixture was stirred at 0 ℃ for 0.5 hour, and PhSO was added 2 Cl (21.78g, 123.34mmol). After the reaction mixture was stirred at 0 ℃ for 1.5 hours, the reaction mixture was cooled to 0 ℃. Slowly adding water to quench the reaction, and then adding ethyl acetate and water to dilute. The organic phase was separated and the aqueous phase was extracted with ethyl acetate. Combining the organic phases, washing with saturated aqueous NaCl solution and anhydrous Na 2 SO 4 Drying, filtration and concentration under reduced pressure gave the title compound D-1 (17.50g, 94%) as a grey solid. LC-MS (ESI) M/z 303.0 (M + H) + .
Step 2: synthesis of Compound D-2
D-1 (10.00g, 33.08mmol), anhydrous tetrahydrofuran (200 mL) was added to the reaction flask. The mixture was cooled to-78 ℃ under nitrogen and LDA (49.62mL, 99.24mmol) was added dropwise slowly. After dropping, stirring at-78 deg.C for 1 hr, slowly dropping I 2 (12.59g, 49.62mmol) in tetrahydrofuran (40 mL). After the reaction mixture was stirred at-78 ℃ for 1 hour, the reaction was quenched by slowly adding saturated aqueous ammonium chloride. After the reaction solution was warmed to room temperature, ethyl acetate and water were added to dilute the reaction solution. The organic phase was separated and the aqueous phase was extracted with ethyl acetate. Combining the organic phases, washing with saturated aqueous NaCl solution and anhydrous Na 2 SO 4 Drying, filtering, and concentrating under reduced pressure to obtain crude target compound D-2 as a reddish brown solid (17.00 g,>99%) and the crude solid was used directly in the next reaction.
And step 3: synthesis of Compound D-3
To the reaction flask was added D-2 (14.16 g (theory), 33.08 mmol), methanol (100 mL), tetrahydrofuran (50 mL), potassium carbonate (13.71mL, 99.21mmol). The mixture was stirred at 80 ℃ for 2 hours, then cooled to room temperature and concentrated under reduced pressure to give the crude product. The crude product was diluted with ethyl acetate and water. The organic phase was separated and the aqueous phase was extracted with ethyl acetate. The combined organic phases were washed with saturated aqueous NaCl solution and concentrated under reduced pressure, and the crude product was purified by automatic column chromatography (Biotage) (mobile phase: petroleum ether/ethyl acetate 100/0 to 80/20) to give the title compound D-3 as a brown solid (1.49g, 16%). LC-MS (ESI) M/z 288.9 (M + H) + .
Step 4 Synthesis of Compound D-4
D-3 (1.49g, 5.17mmol), anhydrous tetrahydrofuran (30 mL) was added to the reaction flask. The mixture was cooled to 0 ℃ under nitrogen and NaH (1.03g, 25.86mmol) was added. After the addition, the temperature is raised to room temperature and the mixture is stirred for 0.5 hour, CF is added 3 CH 2 OTf (4.80g, 20.69mmol). After the reaction mixture was stirred at room temperature for 2 hours, the reaction mixture was cooled to 0 ℃. Slowly adding water to quench the reaction, and then adding ethyl acetate and water to dilute. The organic phase was separated and the aqueous phase was extracted with ethyl acetate. Combining the organic phases, washing with saturated aqueous NaCl solution and anhydrous Na 2 SO 4 Drying, filtration and concentration under reduced pressure gave the title compound D-4 (1.16g, 61%) as a brown solid. LC-MS (ESI): M/z371.0 (M + H) + .
Step 5 Synthesis of Compound D-5
Add D-4 (1.16g, 3.13mmol), ethanol (45 mL), saturated NH to the reaction flask 4 Aqueous Cl (15 mL), iron powder (1.05g, 18.81mmol). The mixture was stirred at 70 ℃ for 1 hour and then cooled to room temperature. After most of ethanol was removed by concentration under reduced pressure, ethyl acetate and water were added to dilute. The organic phase was separated and the aqueous phase was extracted with ethyl acetate. The combined organic phases were washed with saturated aqueous NaCl solution and concentrated under reduced pressure, and the crude product was purified by automatic column chromatography (Biotage) (mobile phase: petroleum ether/ethyl acetate 100/0 to 86/14) to give the title compound D-5 (0.71g, 66%) as a brown solid. LC-MS (ESI) M/z 341.0 (M + H) + .
Step 6 Synthesis of Compound D-6
Into a reaction flaskD-5 (850mg, 2.50mmol), tert-butyl 3-fluoro-4-oxopiperidine-1-carboxylate (2.71g, 12.50mmol), acetic acid (15 mL), 1, 2-dichloroethane (5 mL), sodium triacetoxyborohydride (1.32g, 6.25mmol) were added. The mixture was stirred at 40 ℃ for 2 hours under nitrogen and then cooled to room temperature. The mixture was quenched with water, adjusted to pH 9-10 with 2M aqueous sodium hydroxide, and diluted with ethyl acetate and water. The organic phase was separated and the aqueous phase was extracted with ethyl acetate. The combined organic phases were washed with saturated aqueous NaCl solution and concentrated under reduced pressure, and the crude product was purified by an automatic column chromatography (Biotage) (mobile phase: petroleum ether/ethyl acetate 100/0 to 90/10) to give the desired compound D-6 (1.18g, 87%) as a brown solid. LC-MS (ESI): M/z 542.0 (M + H) + .
Step 7 Synthesis of Compound D-7
To the reaction flask were added D-6 (1.10g, 2.03mmol), dichloromethane (10 mL), and trifluoroacetic acid (5 mL). After the mixture was stirred at room temperature for 1 hour, the reaction was quenched by slowly adding saturated aqueous sodium carbonate solution. The resulting mixture was diluted with ethyl acetate and water, the organic phase was separated and the aqueous phase was extracted with ethyl acetate. Mixing the organic phases, washing with saturated aqueous NaCl solution, and removing anhydrous Na 2 SO 4 Drying, filtration and concentration under reduced pressure gave compound D-7 as a pale yellow solid (870mg, 97%). LC-MS (ESI) M/z 441.9 (M + H) + .
Step 8 Synthesis of Compound D
To the reaction flask were added D-7 (850mg, 1.93mmol), paraformaldehyde (289mg, 9.63mmol), methanol (20 mL), acetic acid (116mg, 1.93mmol), and sodium cyanoborohydride (242mg, 3.85mmol). The mixture was stirred at 50 ℃ for 1 hour under nitrogen and then cooled to room temperature. The mixture was slowly added to a saturated aqueous sodium carbonate solution and stirred for 0.5 hours. Ethyl acetate and water were added. The organic phase was separated and the aqueous phase was extracted with ethyl acetate. The combined organic phases were washed with saturated aqueous NaCl and concentrated under reduced pressure, and the crude product was purified by automatic column chromatography (Biotage) (mobile phase: petroleum ether/ethyl acetate 100/0 to 0/100) to give an off-white solid (756mg, 86%). The crude product was purified by Pre-TLC to give intermediate D (developer: petroleum ether/tetrahydrofuran 1/1), LC-MS (ESI): M/z 456.0 (M + H) + 。 1 H NMR(400MHz,DMSO-d 6 )δ7.26(s,1H),6.97–6.80(m,2H),6.24(d,J＝7.7Hz,1H),5.41(d,J＝8.7Hz,1H),5.00(q,J＝9.0Hz,2H),4.81(d,J＝49.3Hz,1H),3.67–3.51(m,1H),3.12–2.99(m,1H),2.81(d,J＝11.3Hz,1H),2.31–2.15(m,4H),2.14–2.05(m,1H),1.99–1.86(m,1H),1.76–1.66(m,1H).
Synthetic route to example 1
Step 1: synthesis of Compound 1-1
To the reaction flask was added 2-methoxy-4-methylsulphonyl-aniline (1.12g, 5.57mmol), (3-bromomethylphenyl) boronic acid pinacol ester (3.31g, 11.13mmol), DMF (15 mL), caesium carbonate (5.44g, 16.70mmol) and heated to 100 ℃ and stirred overnight. The next day, the reaction solution was cooled to room temperature and diluted with ethyl acetate and water. The organic phase was separated and the aqueous phase was extracted with ethyl acetate. Combining the organic phases, washing with saturated aqueous NaCl solution and anhydrous Na 2 SO 4 Drying, filtration, concentration under reduced pressure and purification by autosampler (Biotage) (mobile phase: petroleum ether/ethyl acetate 100/0 to 80/20) afforded compound 1-1 as a white solid (440mg, 19%). LC-MS (ESI) M/z 418.0 (M + H) + 。 1 H NMR (400 MHz, deuterated chloroform) δ 7.79 (s, 1H), 7.75 (dt, J =7.3,1.3hz, 1h), 7.47-7.39 (m, 2H), 7.37 (t, J =7.4hz, 1h), 7.23 (d, J =2.0hz, 1h), 6.60 (d, J =8.4hz, 1h), 4.40 (s, 2H), 3.91 (s, 3H), 3.01 (s, 3H), 1.35 (s, 12H).
Step 2: synthesis of Compound 1
Add intermediate B (156mg, 335. Mu. Mol), compound 1-1 (215mg, 515. Mu. Mol), DMF (10 mL), water (2 mL), potassium carbonate (105mg, 759. Mu. Mol) to the reaction flask, replace 3 to 5 times with nitrogen, add Pd (PPh) 3 ) 4 (18mg, 0.015mmol), and the nitrogen gas was replaced 3 to 5 times. The mixture was stirred at 100 ℃ for 2 hours under nitrogen and then cooled to room temperature. After adding distilled water and stirring at room temperature for an additional 1 hour, ethyl acetate and water were added for dilution. The organic phase was separated and the aqueous phase was extracted with ethyl acetate. Combining the organic phases, washing with saturated aqueous NaCl solution and anhydrous Na 2 SO 4 Drying, filtering, concentrating under reduced pressure to obtain crude product of compound 1,compound 1 was isolated and purified to give target compound 1-p1 (8mg, 5%) as a white solid and target compound 1-p2 (31mg, 14%) as a white solid. LC-MS (ESI): M/z629.1 (M + H) + . The preparation method comprises the following steps: boston pHlex ODS, 21.2X 250mm,10um, water (0.05% trifluoroacetic acid)/acetonitrile, flow rate of 30mL/min, column temperature of 25 ℃, detection wavelength of 254 nm).
Compound 1-p1 (8 mg, 5%) spectral data: 1 H NMR(400MHz,DMSO-d 6 )δ7.47–7.42(m,2H),7.40–7.33(m,2H),7.26–7.17(m,3H),6.97(t,J＝8.0Hz,1H),6.83(d,J＝8.1Hz,2H),6.73(t,J＝6.4Hz,1H),6.54(d,J＝8.4Hz,1H),6.19(d,J＝7.7Hz,1H),5.40–5.30(m,2H),5.02–4.94(m,2H),4.50(d,J＝6.3Hz,2H),3.90(s,3H),3.04(s,3H),2.21(s,6H),2.08(d,J＝12.1Hz,2H),2.00(q,J＝7.0,6.5Hz,2H),1.85(d,J＝12.1Hz,2H).
compound 1-p2 (31mg, 14%) spectral data: 1 h NMR (400 MHz, deuterated chloroform) δ 7.48-7.42 (m, 2H), 7.41-7.33 (m, 2H), 7.24-7.18 (m, 2H), 7.00-6.93 (m, 2H), 6.83 (d, J =8.3hz, 1h), 6.73 (t, J =6.4hz, 1h), 6.54 (d, J =8.4hz, 1h), 6.20 (d, J =7.8hz, 1h), 5.31 (d, J =7.7hz, 1h), 4.97 (q, J =8.9hz, 2h), 4.50 (d, J =6.3hz, 2h), 3.90 (s, 3H), 3.04 (s, 3H), 2.16 (s, 6H), 2.00 (dd, J =18.0,10.4hz, 2h), 1.77 (d, J =11.5hz, 4h), 1.64 (s, 2H).
EXAMPLE 2 Synthesis of Compound 2
Step 1: synthesis of Compound 2-1
To the reaction flask was added 4-bromomethylphenylboronic acid pinacol ester (700mg, 2.36mmol), 2-methoxy-4-methylsulfonyl-aniline (569mg, 2.83mmol), DMF (10 mL), cesium carbonate (920mg, 2.83mmol), and the mixture was heated to 100 ℃ and stirred overnight. The next day, the reaction solution was cooled to room temperature and diluted with ethyl acetate and water. The organic phase was separated and the aqueous phase was extracted with ethyl acetate. The combined organic phases were washed with saturated aqueous sodium chloride solution, concentrated under reduced pressure and purified by an automatic column chromatography (Biotage) (mobile phase: petroleum ether/ethyl acetate 100/0 to 80/20) to give compound 2-1 as a white solid (145 mg,15％)。LC-MS(ESI):m/z 418.2(M+H) + .
step 2: synthesis of Compound 2
Into a reaction flask were charged compound B (150mg, 0.32mmol), compound 2-1 (270mg, 0.39mmol), potassium carbonate (89mg, 0.64mmol), DMF (10 mL), water (2 mL), and Pd (PPh) was added after 3 to 5 nitrogen replacements 3 ) 4 (37mg, 0.03mmol), and the nitrogen was further replaced 3 to 5 times. The mixture was stirred at 100 ℃ for 2 hours under nitrogen. After cooling to room temperature, ethyl acetate and water were added for dilution. The organic phase was separated and the aqueous phase was extracted with ethyl acetate. And (3) combining organic phases, washing with saturated sodium chloride aqueous solution, drying with anhydrous sodium sulfate, filtering, concentrating under reduced pressure, and preparing, separating and purifying an obtained crude product to obtain a white solid, namely a compound 2:
compound 2 was purified to give the single isomer (syn or anti) target compound 2-p1 (17mg, 8%) and the white solid single isomer (syn or anti) target compound 2-p2 (22mg, 11%).
The preparation method comprises the following steps: boston pHlex ODS, 21.2X 250mm,10um, water (0.05% trifluoroacetic acid)/acetonitrile, flow rate of 30mL/min, column temperature of 25 ℃, detection wavelength of 254 nm).
Compound 2-p1 spectral data: LC-MS (ESI): M/z629.1 (M + H) + 。 1 H NMR(400MHz,DMSO-d 6 )δ7.44(d,J＝1.2Hz,4H),7.29–7.20(m,2H),6.97(t,J＝8.0Hz,1H),6.89–6.80(m,2H),6.71(t,J＝6.3Hz,1H),6.56(d,J＝8.5Hz,1H),6.19(d,J＝7.8Hz,1H),5.33(d,J＝7.6Hz,1H),5.00(q,J＝8.7Hz,2H),4.50(d,J＝6.3Hz,2H),3.93(s,3H),3.22(s,1H),3.07(s,3H),2.21(s,6H),2.08(d,J＝12.1Hz,2H),2.00(q,J＝7.0,6.6Hz,1H),1.85(d,J＝11.9Hz,2H),1.30(t,J＝10.8Hz,4H).
Compound 2-p2 spectral data: LC-MS (ESI) M/z629.1 (M + H) + 。 1 H NMR(400MHz,DMSO-d 6 )δ7.45(s,4H),7.32–7.18(m,2H),7.05–6.89(m,2H),6.84(t,J＝9.8Hz,1H),6.71(t,J＝6.3Hz,1H),6.56(d,J＝8.5Hz,1H),6.20(d,J＝7.7Hz,1H),5.26(d,J＝7.5Hz,1H),5.01(q,J＝9.0Hz,2H),4.50(d,J＝6.3Hz,2H),3.93(s,3H),3.52(d,J＝12.9Hz,1H),3.07(s,3H),2.18(s,6H),2.08(s,1H),1.77(d,J＝9.5Hz,4H),1.64(s,2H),1.50(s,2H).
EXAMPLE 3 Synthesis of Compound 3
Synthesis of Compound 3
Add intermediate D (70mg, 0.15mmol), intermediate 1-1 (95mg, 0.23mmol), DMF (5 mL), water (1 mL), potassium carbonate (43mg, 0.31mmol) to the reaction flask, replace 3 to 5 times with nitrogen, add Pd (PPh) 3 ) 4 (18mg, 0.015mmol), and the nitrogen gas was replaced 3 to 5 times. The mixture was stirred at 100 ℃ for 2 hours under nitrogen and then cooled to room temperature. After addition of saturated aqueous EDTA and stirring at room temperature for a further 1 hour, it is diluted with ethyl acetate and water. The organic phase was separated and the aqueous phase was extracted with ethyl acetate. The organic phases were combined, washed with saturated aqueous sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure, and the crude product was isolated and purified to give the title compound 3 as a pale yellow solid, and the compound 3 was prepared to give the compound 3-p1 (60mg, 63%) and 3-p2 (12mg, 29%).
The preparation method comprises the following steps: boston pHlex ODS, 21.2X 250mm,10um, water (0.05% trifluoroacetic acid)/acetonitrile, flow rate of 30mL/min, column temperature of 25 ℃, detection wavelength of 254 nm),
compound 3-p1 spectral data: LC-MS (ESI) M/z 619.2 (M + H) + 。 1 H NMR(400MHz,DMSO-d 6 )δ7.50–7.41(m,2H),7.41–7.34(m,2H),7.25–7.17(m,2H),6.99(t,J＝7.9Hz,1H),6.96–6.87(m,2H),6.73(t,J＝6.4Hz,1H),6.54(d,J＝8.4Hz,1H),6.28(d,J＝7.6Hz,1H),5.38(d,J＝8.7Hz,1H),4.99(q,J＝9.0Hz,2H),4.84(d,J＝49.4Hz,1H),4.51(d,J＝6.4Hz,2H),3.91(s,3H),3.69–3.49(m,1H),3.11–2.99(m,4H),2.81(d,J＝11.2Hz,1H),2.31–2.16(m,4H),2.14–2.06(m,1H),1.99–1.86(m,1H),1.77–1.68(m,1H).
Compound 3-p2 spectral data: LC-MS (ESI): M/z 619.2 (M + H) + 。 1 H NMR(400MHz,DMSO-d 6 )δ7.50–7.42(m,2H),7.42–7.33(m,2H),7.25–7.17(m,2H),6.97(t,J＝7.9Hz,1H),6.90–6.81(m,2H),6.74(t,J＝6.4Hz,1H),6.54(d,J＝8.4Hz,1H),6.27(d,J＝7.7Hz,1H),5.64(d,J＝8.3Hz,1H),4.98(q,J＝9.0Hz,2H),4.68–4.45(m,3H),3.91(s,3H),3.61–3.46(m,1H),3.15–2.99(m,4H),2.75–2.64(m,1H),2.24(s,3H),2.15–2.08(m,1H),2.07–1.98(m,2H),1.52–1.43(m,1H).
Example 4: synthetic route to Compound 4
Step 1: synthesis of Compound 4-1
To the reaction flask was added 2-methoxy-4-methylsulfonyl-aniline (2.00g, 9.94mmol), di-tert-butyl dicarbonate (13.01g, 59.63mmol), 4-dimethylaminopyridine (1.46g, 11.93mmol), 1,4-dioxane (30 mL). The mixture was heated to 110 ℃ and stirred overnight. The next day, the reaction solution was cooled to room temperature and diluted with ethyl acetate and water. The organic phase was separated and the aqueous phase was extracted with ethyl acetate. And (3) combining organic phases, washing with saturated sodium chloride aqueous solution, drying with anhydrous sodium sulfate, filtering, and concentrating under reduced pressure to obtain a crude product. To the crude product was added potassium carbonate (4.12g, 29.82mmol), methanol (30 mL) and the mixture was heated to 40 ℃ and stirred for 6 hours. After cooling to room temperature, the mixture is concentrated under reduced pressure, the concentrate obtained is extracted with ethyl acetate and water, the organic phase is separated and the aqueous phase is extracted with ethyl acetate. The combined organic phases were washed with saturated aqueous sodium chloride and the crude product was purified by automatic column chromatography (Biotage) (mobile phase: petroleum ether/ethyl acetate 100/0 to 50/50) to give compound 4-1 as a white solid (2.15g, 72%). 1 H NMR (400 MHz, deuterated chloroform) δ 8.30 (d, J =8.6hz, 1h), 7.53 (dd, J =8.6,2.0hz, 1h), 7.36 (d, J =2.0hz, 1h), 7.30 (s, 1H), 3.95 (s, 3H), 3.03 (s, 3H), 1.53 (s, 9H).
Step 2: synthesis of Compound 4-2
To the reaction flask were added compound 4-1 (500mg, 1.66mmol), 3-bromobenzyl bromide (0.83g, 3.32mmol), cesium carbonate (1.62g, 4.98mmol), and DMF (8 mL). After the mixture was heated to 50 ℃ and stirred for 2 hours, the reaction solution was cooled to room temperature and diluted with ethyl acetate and water. The organic phase was separated and the aqueous phase was extracted with ethyl acetate. Mixing organic phases, washing with saturated sodium chloride aqueous solution, concentrating under reduced pressure, and subjecting the obtained crude product to automatic column chromatography (Bi)otage) purification (mobile phase: petroleum ether/ethyl acetate 100/0 to 65/35) to give compound 4-2 (780 mg, 100%) as a white solid. 1 H NMR (400 MHz, deuterated chloroform) delta 7.50-7.39 (m, 3H), 7.38-7.31 (m, 1H), 7.24-7.07 (m, 3H), 4.67 (s, 2H), 3.87 (s, 3H), 3.04 (s, 3H), 1.40 (s, 9H).
And 3, step 3: synthesis of Compound 4-3
2-Nitrocarbazole (110mg, 0.518mmol), compound 4-2 (293mg, 0.622mmol), potassium carbonate (215mg, 1.56mmol), cuprous iodide (99mg, 0.518mmol), DMF (10 mL) were charged into a reaction flask, and after 3-5 nitrogen replacements, N' -dimethylethylenediamine (5mg, 0.052mmol) was added, and nitrogen replacements were further performed 3-5 times. After the mixture was heated to 130 ℃ and stirred overnight, the reaction solution was cooled to room temperature and diluted with ethyl acetate and water. The organic phase was separated and the aqueous phase was extracted with ethyl acetate. The combined organic phases were washed with saturated aqueous sodium chloride solution and concentrated under reduced pressure, and the crude product was purified by automatic column chromatography (Biotage) (mobile phase: petroleum ether/ethyl acetate 100/0 to 60/40) to give compound 4-3 as a yellow solid (189mg, 61%). LC-MS (ESI) at M/z 619.3 (M + NH) 4 ) + .
And 4, step 4: synthesis of Compound 4-4
To the reaction flask were added compound 4-3 (180mg, 0.299mmol), ethanol (12 mL), saturated aqueous ammonium chloride (4 mL), and iron powder (100mg, 1.79mmol). The mixture was heated to 70 ℃ and stirred for 1 hour, then cooled to room temperature and diluted with ethyl acetate and water. The organic phase was separated and the aqueous phase was extracted with ethyl acetate. The combined organic phases were washed with saturated aqueous sodium chloride and concentrated under reduced pressure, and the crude product was purified by autosampler (Biotage) (mobile phase: petroleum ether/ethyl acetate 100/0 to 50/50) to give compound 4-4 as a yellow solid (120mg, 70%). LC-MS (ESI) M/z 572.2 (M + H) + .
And 5: synthesis of Compounds 4 to 5
To a reaction flask, compound 4-4 (60mg, 0.105mmol), 3-fluoro-4-oxopiperidine-1-carboxylic acid tert-butyl ester (114mg, 0.524mmol), acetic acid (1.5 mL), 1, 2-dichloroethane (0.5 mL), sodium triacetoxyborohydride (56mg, 0.262mmol) were added. The mixture was stirred at 40 ℃ for 2 hours under nitrogen and then cooled to room temperature. The mixture was quenched with water and dissolved in 2M sodium hydroxideAdjusting pH to 9-10, adding ethyl acetate and water to dilute. The organic phase was separated and the aqueous phase was extracted with ethyl acetate. The combined organic phases were washed with saturated aqueous sodium chloride solution and concentrated under reduced pressure, and the crude product was purified by automatic column chromatography (Biotage) (mobile phase: petroleum ether/ethyl acetate 100/0 to 60/40) to give the title compound 4-5 (60mg, 74%) as a white solid. LC-MS (ESI) M/z 773.1 (M + H) + .
Step 6: synthesis of Compounds 4-6
To the reaction flask were added 4-5 (60mg, 0.077mmol), dichloromethane (6 mL), trifluoroacetic acid (3 mL). After the mixture was stirred at room temperature for 1 hour, the reaction was quenched by slowly adding saturated aqueous sodium carbonate solution. The resulting mixture was diluted with ethyl acetate and water, the organic phase was separated and the aqueous phase was extracted with ethyl acetate. The combined organic phases were washed with saturated aqueous sodium chloride solution, dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to give compound 4-6 as a pale yellow solid (43mg, 97%). LC-MS (ESI) M/z 573.1 (M + H) + .
And 7: synthesis of Compound 4
To the reaction flask were added compound 4-6 (43mg, 0.075mmol), paraformaldehyde (11mg, 0.373mmol), methanol (5 mL), acetic acid (5mg, 0.075mmol), sodium cyanoborohydride (14mg, 0.225mmol). The mixture was stirred at 50 ℃ for 1 hour under nitrogen and then cooled to room temperature. The mixture was slowly added to a saturated aqueous sodium carbonate solution and stirred for 0.5 hours. Ethyl acetate and water were added. The organic phase was separated and the aqueous phase was extracted with ethyl acetate. And combining organic phases, washing with saturated sodium chloride aqueous solution, and concentrating under reduced pressure to obtain a crude product, namely a compound 4, wherein the preparation method for purifying the compound 4 comprises the following steps: boston pHlex ODS, 21.2X 250mm,10um, water (0.05% trifluoroacetic acid)/acetonitrile, flow rate 30mL/min, column temperature 25 ℃, detection wavelength 254 nm), yielded a white solid single isomer (R or S) target compound 4-p1 (15mg, 34%) and a white solid single isomer (R or S) target compound 4-p2 (5mg, 11%).
Compound 4-p1: LC-MS (ESI) M/z 587.3 (M + H) + 。 1 H NMR(400MHz,DMSO-d 6 )δ7.98–7.91(m,1H),7.86(d,J＝8.4Hz,1H),7.60(t,J＝7.7Hz,1H),7.51(s,1H),7.44(d,J＝7.7Hz,2H),7.29(dd,J＝8.3,2.0Hz,1H),7.21(s,1H),7.17–7.10(m,2H),7.10–7.03(m,1H),6.78(t,J＝6.4Hz,1H),6.70(d,J＝8.2Hz,1H),6.62(d,J＝6.7Hz,2H),5.73(d,J＝8.8Hz,1H),4.88–4.67(m,1H),4.57(d,J＝6.4Hz,2H),3.88(s,3H),3.55–3.42(m,1H),3.09–2.94(m,4H),2.74(d,J＝11.1Hz,1H),2.29–2.13(m,4H),2.09–2.00(m,1H),1.78–1.65(m,2H)。
Compound 4-p2: LC-MS (ESI) M/z 587.3 (M + H) + 。 1 H NMR(400MHz,DMSO-d 6 )δ7.98–7.89(m,1H),7.85(d,J＝8.4Hz,1H),7.60(t,J＝7.8Hz,1H),7.50(s,1H),7.47–7.39(m,2H),7.29(d,J＝8.4Hz,1H),7.21(s,1H),7.17–7.09(m,2H),7.08–7.01(m,1H),6.78(t,J＝6.7Hz,1H),6.63(dd,J＝15.0,8.4Hz,2H),6.56(s,1H),5.89(d,J＝8.1Hz,1H),4.57(d,J＝6.4Hz,2H),4.52–4.29(m,1H),3.88(s,3H),3.45–3.41(m,1H),3.06(s,3H),3.02–2.94(m,1H),2.67–2.60(m,1H),2.20(s,3H),2.17–2.11(m,1H),2.05–1.95(m,2H),1.46–1.37(m,1H)。
EXAMPLE 5 synthetic route to Compound 5
Step 1: synthesis of Compound 5-1
To the reaction flask were added compound 4-4 (43mg, 0.075mmol), absolute ethanol (3 mL), tetrahydrothiopyran-4-one 1, 1-dioxide (33mg, 0.225mmol), ethyl titanate (86mg, 0.376 mmol). After stirring the mixture overnight at 50 ℃ under nitrogen, sodium cyanoborohydride (47mg, 0.752mmol) was added and the mixture was warmed to 80 ℃ and stirred for 2 hours. After the reaction, ethyl acetate and water were added to dilute. The organic phase was separated and the aqueous phase was extracted with ethyl acetate. The combined organic phases were washed with saturated aqueous sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to give the desired compound 5-1 (73 mg,>99％)。LC-MS(ESI):m/z 704.2(M+H) + .
step 2: synthesis of Compound 5
Add 5-1 (53mg, 0.075mmol), dichloromethane (4.5 mL), trifluoroacetic acid (2 mL) to the reaction flask. After the mixture was stirred at room temperature for 1 hour, the reaction was quenched by slowly adding saturated aqueous sodium carbonate solution. What is needed isThe resulting mixture was diluted with ethyl acetate and water, the organic phase was separated and the aqueous phase was extracted with ethyl acetate. The combined organic phases were washed with saturated aqueous sodium chloride, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to afford the crude product as a crude purified compound 5 as a white solid (25mg, 55%). LC-MS (ESI): M/z 604.2 (M + H) + 。 1 H NMR(400MHz,DMSO-d 6 )δ7.97–7.92(m,1H),7.89(d,J＝8.4Hz,1H),7.61(t,J＝7.7Hz,1H),7.52–7.41(m,3H),7.29(dd,J＝8.4,2.0Hz,1H),7.20(d,J＝2.0Hz,1H),7.18–7.10(m,2H),7.09–7.03(m,1H),6.78(t,J＝6.4Hz,1H),6.67–6.59(m,2H),6.55(d,J＝1.9Hz,1H),5.92(d,J＝8.6Hz,1H),4.57(d,J＝6.4Hz,2H),3.87(s,3H),3.77–3.64(m,1H),3.25–3.11(m,4H),3.06(s,3H),2.23–2.13(m,2H),1.98–1.86(m,2H).
EXAMPLE 6 synthetic route to Compound 6
Step 1: synthesis of Compound 6-1
To a reaction flask was added (N-tert-butoxycarbonyl-N-methylamino) aniline (833mg, 4mmol), potassium carbonate (533mg, 4mmol), DMF (10 mL), 4-bromomethylbenzeneboronic acid pinacol ester (594mg, 2mmol). N for the mixture 2 The mixture was replaced and stirred at 70 ℃ overnight. The next day the reaction was cooled to room temperature and diluted with ethyl acetate and water. The organic phase was separated and the aqueous phase was extracted with ethyl acetate. The organic phases were combined, washed with saturated aqueous sodium chloride solution and concentrated under reduced pressure. The crude product obtained was purified by automatic column chromatography (Biotage) (mobile phase: petroleum ether/ethyl acetate 100/0 to 90/10) to yield the desired compound 6-1 as a white solid (425mg, 48%). LC-MS (ESI) M/z 425.4 (M + H) + .
And 2, step: synthesis of Compound 6
Add Compound B (173mg, 0.37mmol), compound 6-1 (190mg, 0.45mmol), DMF (5 mL), water (1 mL), potassium carbonate (103mg, 0.75mmol) to the reaction flask, replace 3 to 5 times with nitrogen, add Pd (PPh) 3 ) 4 (124mg, 0.11mmol), and the nitrogen gas was further replaced 3 to 5 times. The mixture was stirred at room temperature for 2 hours under nitrogen, and then addedSaturated aqueous EDTA was added and stirring continued at room temperature for 1 hour. Add ethyl acetate and water to dilute. The organic phase was separated and the aqueous phase was extracted with ethyl acetate. The combined organic phases were washed with saturated aqueous sodium chloride solution and concentrated under reduced pressure, and the resulting crude product was purified by an automatic column chromatography (Biotage) (mobile phase: dichloromethane/methanol 100/0 to 90/10) to give a yellow solid compound (252mg, 78%), which was dissolved in trifluoroacetic acid (2 mL) and dichloromethane (1 mL), stirred at room temperature for 2 hours, neutralized to weak alkalinity with saturated sodium bicarbonate, the aqueous phase was extracted with ethyl acetate, the combined organic phases were spin-dried to give a crude product, compound 6, which was prepared by the following method after purification by prep-HPLC (106 mg): boston pHlex ODS,21.2 x 250mm,10um, water (0.05% trifluoroacetic acid)/acetonitrile, flow rate 30mL/min, column temperature 25 ℃, detection wavelength 254 nm), and the preparation was basified with saturated sodium bicarbonate to give a single isomer (Syn or Anti) compound 6-p1 (10mg, 7%) and a single isomer (Syn or Anti) compound 6-p2 (10mg, 7%).
Compound 6-p1: LC-MS (ESI) M/z 536.0 (M + H) + 。 1 H NMR(400MHz,DMSO-d 6 )δ7.52–7.26(m,4H),7.03–6.94(m,1H),6.83(d,J＝9.6Hz,2H),6.68(t,J＝7.9Hz,1H),6.19(d,J＝7.9Hz,1H),5.94(t,J＝6.2Hz,1H),5.87(t,J＝2.1Hz,1H),5.86–5.71(m,2H),5.55–5.26(m,1H),5.06–4.88(m,2H),4.68(s,2H),4.26(d,J＝6.2Hz,2H),2.18(s6H),1.46–1.02(m,10H).
Compound 6-p2: LC-MS (ESI): M/z 536.0 (M + H) + 。 1 H NMR(400MHz,DMSO-d 6 )δ7.64–7.16(m,4H),7.09–6.88(m,2H),6.84(s,1H),6.67(d,J＝7.8Hz,1H),6.20(d,J＝7.8Hz,1H),5.93(t,J＝6.2Hz,1H),5.86(d,J＝2.1Hz,1H),5.81(td,J＝7.7,2.0Hz,2H),5.31(d,J＝7.8Hz,1H),4.96(s,2H),4.68(s,2H),4.27(d,J＝6.1Hz,2H),2.17(s,6H),2.09–1.89(m,2H),1.77(d,J＝9.5Hz,4H),1.65(dd,J＝8.8,4.0Hz,2H),1.55–1.37(m,2H).
EXAMPLE 7 synthetic route to Compound 7
Step 1: synthesis of Compound 7-1
To the reaction flask were added 3- (N-tert-butoxycarbonyl-N-methylamino) aniline (222mg, 2mmol), potassium carbonate (276 mg, 2mmol), DMF (10 mL), 4-bromomethylbenzeneboronic acid pinacol ester (297 mg, 1mmol). From the mixture with N 2 The mixture was replaced and stirred at 70 ℃ overnight. The next day the reaction was cooled to room temperature and diluted with ethyl acetate and water. The organic phase was separated and the aqueous phase was extracted with ethyl acetate. The organic phases were combined, washed with saturated aqueous sodium chloride solution and concentrated under reduced pressure. The resulting crude product was purified by automated column chromatography (Biotage) (mobile phase: petroleum ether/ethyl acetate 100/0 to 90/10) to afford the title compound 7-1 as a white solid (188mg, 43%). LC-MS (ESI): M/z 439.5 (M + H) + .
Step 2: synthesis of Compound 7
Add intermediate B (210mg, 0.48mmol), compound 7-1 (184mg, 0.43mmol), DMF (5 mL), water (1 mL), potassium carbonate (103mg, 0.75mmol) to the reaction flask, replace 3 to 5 times with nitrogen, add Pd (PPh) 3 ) 4 (124mg, 0.11mmol), and the nitrogen gas was further replaced 3 to 5 times. After stirring the mixture at room temperature for 2 hours under nitrogen, saturated aqueous EDTA solution was added and stirring was continued at room temperature for 1 hour. Add ethyl acetate and water to dilute. The organic phase was separated and the aqueous phase was extracted with ethyl acetate. The combined organic phases were washed with saturated aqueous sodium chloride solution and concentrated under reduced pressure, and the crude product was purified by an automatic column chromatography (Biotage) (mobile phase: dichloromethane/methanol 100/0 to 90/10) to give a yellow solid compound (110mg, 78%), which was dissolved in trifluoroacetic acid (2 mL) and dichloromethane (1 mL), stirred at room temperature for 2 hours, neutralized to weak alkalinity with saturated sodium bicarbonate, the aqueous phase was extracted with ethyl acetate, and the combined organic phases were purified by the following method as compound 7, prep-HPLC (106 mg): the preparation method comprises the following steps: boston pHlex ODS,21.2 x 250mm,10um, water (0.05% trifluoroacetic acid)/acetonitrile, flow rate of 30mL/min, column temperature 25 ℃, detection wavelength 254 nm).
Basification of the preparation with saturated sodium bicarbonate yielded the single isomers (Syn or Anti) compound 7-p1 (10mg, 7%) and compound 7-p2 (10mg, 7%).
Compound 7-p1: LC-MS (ESI) M/z 550.0 (M + H) + 。 1 H NMR(400MHz,DMSO-d 6 )δ7.48–7.22(m,4H),6.97(t,J＝7.9Hz,1H),6.83(d,J＝8.2Hz,2H),6.74(t,J＝7.9Hz,1H),6.19(d,J＝7.8Hz,1H),5.99(t,J＝6.2Hz,1H),5.89–5.73(m,3H),5.46–5.29(m,1H),5.24(q,J＝5.0Hz,1H),4.97(q,J＝9.0Hz,2H),4.29(d,J＝6.1Hz,2H)2.56(d,J＝5.0Hz,3H),2.18(s,6H),2.09(s,3H),1.84(d,J＝11.5Hz,2H),1.51–1.24(m,5H).
Compound 7-p2: LC-MS (ESI): M/z 550.0 (M + H) + 。 1 H NMR(400MHz,DMSO-d 6 )δ7.53–7.29(m,4H),7.13–6.91(m,2H),6.84(d,J＝8.2Hz,1H),6.74(t,J＝7.8Hz,1H),6.20(d,J＝7.7Hz,1H),5.99(t,J＝6.1Hz,1H),5.92–5.66(m,3H),5.31(d,J＝7.4Hz,1H),5.24(d,J＝5.3Hz,1H),4.97(q,J＝9.2Hz,2H),4.29(d,J＝6.1Hz,2H),2.56(d,J＝4.6Hz,3H),2.18(s,6H),2.13–1.94(m,2H),1.79(dd,J＝11.7,6.8Hz,4H),1.64(s,4H),1.56–1.41(m,2H).
EXAMPLE 8 Synthesis of Compound 8
Step 1: synthesis of Compound 8-1
To the reaction flask were added intermediate B-2 (200mg, 0.588mmol), anhydrous ethanol (5 mL), tetrahydrothiopyran-4-one 1, 1-dioxide (261mg, 1.76mmol), and ethyl titanate (670mg, 2.94mmol). After stirring the mixture overnight at 50 ℃ under nitrogen, sodium cyanoborohydride (369mg, 5.88mmol) was added and the mixture was warmed to 80 ℃ and stirred for 2 hours. After the reaction, ethyl acetate and water were added to dilute the reaction mixture. The organic phase was separated and the aqueous phase was extracted with ethyl acetate. The combined organic phases were washed with saturated aqueous sodium chloride solution and concentrated under reduced pressure, and the crude product was purified by an automatic column chromatography (Biotage) (mobile phase: petroleum ether/ethyl acetate 100/0 to 50/50) to give the title compound 8-1 (126mg, 45%) as a yellow solid. LC-MS (ESI) M/z 473.0 (M + H) + .
And 2, step: synthesis of Compound 8
Add intermediate 8-1 (60mg, 0.127mmol), intermediate C (64mg, 0.152mmol), DMF (5 mL), water (1 mL), potassium carbonate (35mg, 0.254mmol), nitrogen to the reaction flaskAfter 3 to 5 replacements, pd (PPh) was added 3 ) 4 (15mg, 0.012mmol), and nitrogen was further substituted 3 to 5 times. The mixture was stirred at 100 ℃ for 2 hours under nitrogen and then cooled to room temperature. After addition of saturated aqueous EDTA and stirring at room temperature for a further 1 hour, it is diluted with ethyl acetate and water. The organic phase was separated and the aqueous phase was extracted with ethyl acetate. The combined organic phases were washed with saturated aqueous sodium chloride, dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure and the crude product was purified preparative (Welch Xtimate C18, 21.2 x 250mm,10um, water (10 mM ammonium bicarbonate)/acetonitrile, flow rate 30mL/min, column temperature 25 ℃, detection wavelength 254 nm) to give target compound 8 (25mg, 31%) as a white solid. LC-MS (ESI) M/z 636.1 (M + H) + 。 1 H NMR(400MHz,DMSO-d 6 )δ7.48–7.42(m,2H),7.42–7.33(m,2H),7.24–7.18(m,2H),7.01(t,J＝7.9Hz,1H),6.90(d,J＝8.3Hz,1H),6.87(s,1H),6.73(t,J＝6.4Hz,1H),6.54(d,J＝8.3Hz,1H),6.27(d,J＝7.7Hz,1H),5.65(d,J＝8.6Hz,1H),4.99(q,J＝9.0Hz,2H),4.51(d,J＝6.3Hz,2H),3.91(s,3H),3.88–3.77(m,1H),3.31–3.25(m,2H),3.21–3.11(m,2H),3.04(s,3H),2.29–2.18(m,2H),2.06–1.97(m,2H).
EXAMPLE 9 synthetic route to Compound 9
Step 1: synthesis of Compound 9-1
To the reaction flask, intermediate B-2 (200mg, 0.588mmol), dehydrated ethanol (5 mL), tetrahydro-4H-pyran-4-one (177mg, 1.76mmol), and ethyl titanate (670mg, 2.94mmol) were added. After stirring the mixture overnight at 50 ℃ under nitrogen, sodium cyanoborohydride (369mg, 5.88mmol) was added and the mixture was warmed to 80 ℃ and stirred for 2 hours. After the reaction, ethyl acetate and water were added to dilute. The organic phase was separated and the aqueous phase was extracted with ethyl acetate. The combined organic phases were washed with saturated aqueous sodium chloride solution and concentrated under reduced pressure, and the crude product was purified by an automatic column chromatography (Biotage) (mobile phase: petroleum ether/ethyl acetate 100/0 to 80/20) to give the title compound 9-1 as an off-white solid (203mg, 81%). LC-MS (ESI) M/z 425.0 (M + H) + .
And 2, step: synthesis of Compound 9
To a reaction flask, compound 9-1 (70mg, 0.165mmol), intermediate C (103mg, 0.247mmol), DMF (5 mL), water (1 mL), potassium carbonate (46mg, 0.330mmol) were added, replaced with nitrogen 3 to 5 times, and Pd (PPh) was added 3 ) 4 (19mg, 0.016mmol), and nitrogen was further substituted 3 to 5 times. The mixture was stirred at 100 ℃ for 2 hours under nitrogen and then cooled to room temperature. After addition of saturated aqueous EDTA and stirring at room temperature for a further 1 hour, dilution is carried out with ethyl acetate and water. The organic phase was separated and the aqueous phase was extracted with ethyl acetate. The combined organic phases were washed with saturated aqueous sodium chloride, dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure and the crude product was purified by column autosampler (Biotage) (mobile phase: petroleum ether/ethyl acetate 100/0 to 50/50) to afford title compound 9 (26mg, 27%) as a white solid. LC-MS (ESI): M/z 588.2 (M + H) + . 1 H NMR (400 MHz, deuterated chloroform) δ 7.48-7.42 (m, 2H), 7.41-7.33 (m, 2H), 7.25-7.17 (m, 2H), 6.98 (t, J =8.0hz, 1h), 6.86 (d, J =6.5hz, 2h), 6.73 (t, J =6.3hz, 1h), 6.54 (d, J =8.4hz, 1h), 6.26 (d, J =7.7hz, 1h), 5.48 (d, J =8.1hz, 1h), 4.98 (q, J =9.0hz, 2h), 4.51 (d, J =6.3hz, 2h), 3.96-3.85 (m, 5H), 3.65-3.54 (m, 1H), 3.44 (td, J =11.7, 2H, 3.3.7, 2H), 3.96-3.85 (m, 5H), 3.65-3.54 (m, 1H), 3.44 (td, J =11.7, 11.3.3.3, 2H), 2H, 1H, 2 m-55H).
Example 10: synthetic route to Compound 10
Step 1: synthesis of Compound 10-1
To the reaction flask was added cyclopropanecarboxylic acid (500mg, 5.81mmol), DMF (15 mL), 3-aminomethylphenylboronic acid pinacol ester hydrochloride (1.25g, 4.65mmol), DIEA (2.25g, 17.42mmol), HATU (3.31g, 8.71mmol). The mixture was stirred at room temperature overnight. After the reaction, ethyl acetate and water were added to dilute. The organic phase was separated and the aqueous phase was extracted with ethyl acetate. The combined organic phases were washed with saturated aqueous sodium chloride solution and concentrated under reduced pressure, and the crude product was purified by an automatic column chromatography (Biotage) (mobile phase: petroleum ether/ethyl acetate 100/0 to 80/20) to give the desired compound 10-1 (159) as a yellow solidmg，11％)。LC-MS(ESI):m/z 302.2(M+H) + .
And 2, step: synthesis of Compound 10
Add intermediate 10-1 (100mg, 0.214mmol), intermediate B (78mg, 0.257mmol), DMF (5 mL), water (1 mL), potassium carbonate (59mg, 0.429mmol) to the reaction flask, replace 3 to 5 times with nitrogen, add Pd (PPh) 3 ) 4 (25mg, 0.021mmol), and the nitrogen was replaced 3 to 5 times. The mixture was stirred at 100 ℃ for 2 hours under nitrogen and then cooled to room temperature. After addition of saturated aqueous EDTA and stirring at room temperature for a further 1 hour, it is diluted with ethyl acetate and water. The organic phase was separated and the aqueous phase was extracted with ethyl acetate. The organic phases were combined, washed with saturated aqueous sodium chloride, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to give crude compound 10, which was purified by compound 10 (Boston pHlex ODS, 21.2X 250mm,10um, water (0.05% trifluoroacetic acid)/acetonitrile, flow rate 30mL/min, column temperature 25 ℃, detection wavelength 254 nm) to give target compound 10-p1 (15mg, 14%) as a white solid and target compound 10-p2 (25mg, 23%) as a white solid.
Compound 10-p1: LC-MS (ESI) M/z 513.0 (M + H) + 。 1 H NMR(400MHz,DMSO-d 6 )δ8.64(t,J＝5.9Hz,1H),7.46(t,J＝7.9Hz,1H),7.37(d,J＝6.8Hz,2H),7.30(d,J＝7.6Hz,1H),6.98(t,J＝7.9Hz,1H),6.84(d,J＝9.7Hz,2H),6.20(d,J＝7.8Hz,1H),5.41(d,J＝8.0Hz,1H),5.03(q,J＝9.0Hz,2H),4.36(d,J＝5.9Hz,2H),3.45–3.41(m,1H),2.27–2.20(m,7H),2.13–2.06(m,2H),1.88–1.82(m,2H),1.65–1.58(m,1H),1.37–1.28(m,4H),0.72–0.65(m,4H).
Compound 10-p2: LC-MS (ESI) M/z 513.0 (M + H) + 。 1 H NMR(400MHz,DMSO-d 6 )δ8.64(t,J＝5.9Hz,1H),7.46(t,J＝7.9Hz,1H),7.38(dd,J＝6.7,1.5Hz,2H),7.30(dd,J＝7.5,1.6Hz,1H),7.01–6.95(m,2H),6.85(d,J＝8.2Hz,1H),6.21(d,J＝7.7Hz,1H),5.33(d,J＝7.3Hz,1H),5.03(q,J＝9.0Hz,2H),4.36(d,J＝5.9Hz,2H),3.59–3.51(m,1H),2.19(s,6H),2.14–2.05(m,1H),1.87–1.73(m,4H),1.69–1.58(m,3H),1.56–1.46(m,2H),0.74–0.63(m,4H).
EXAMPLE 11 Synthesis of Compound 11
Step 1: synthesis of Compound 11-1
To the reaction flask was added 1-methyl-1H-pyrrole-3-carboxylic acid (300mg, 2.40mmol), DMF (10 mL), 3-aminomethylphenylboronic acid pinacol ester hydrochloride (517mg, 1.92mmol), DIEA (929mg, 7.19mmol), HATU (1.37g, 3.60mmol). The mixture was stirred at room temperature overnight. After the reaction, ethyl acetate and water were added to dilute the reaction mixture. The organic phase was separated and the aqueous phase was extracted with ethyl acetate. The combined organic phases were washed with saturated aqueous sodium chloride solution and concentrated under reduced pressure, and the crude product was purified by an automatic column chromatography (Biotage) (mobile phase: petroleum ether/ethyl acetate 100/0 to 70/30) to give the title compound 11-1 (263mg, 40%) as a yellow solid. LC-MS (ESI): M/z 341.2 (M + H) + .
Step 2: synthesis of Compound 11
Add intermediate 11-1 (120mg, 0.257mmol), intermediate B (105mg, 0.309mmol), DMF (5 mL), water (1 mL), potassium carbonate (71mg, 0.515mmol) to the reaction flask, replace 3 to 5 times with nitrogen, add Pd (PPh) 3 ) 4 (30mg, 0.025mmol), and the nitrogen gas was further replaced 3 to 5 times. The mixture was stirred at 100 ℃ for 2 hours under nitrogen and then cooled to room temperature. After addition of saturated aqueous EDTA and stirring at room temperature for a further 1 hour, it is diluted with ethyl acetate and water. The organic phase was separated and the aqueous phase was extracted with ethyl acetate. The organic phases are combined, washed with saturated aqueous sodium chloride solution, dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to obtain the crude compound 11.
Compound 11 was purified by preparative method as follows: (Boston pHlex ODS, 21.2X 250mm,10um, water (0.05% trifluoroacetic acid)/acetonitrile, flow rate of 30mL/min, column temperature of 25 ℃, detection wavelength of 254 nm), to give target compound 11-p1 (13mg, 9%) as a white solid and target compound 11-p2 (15mg, 11%) as a white solid.
Compound 11-p1 spectral data: LC-MS (ESI) M/z 552.0 (M + H) + 1H NMR (400 MHz, deuterated chloroform) δ 8.36 (t, J =6.1hz, 1h), 7.44 (t, J =7.6hz, 1h), 7.39-7.30 (m, 3H), 7.27 (t, J =2.0hz, 1h), 6.97 (t, J =8.0hz, 1h)),6.87–6.80(m,2H),6.69(t,J＝2.5Hz,1H),6.47(dd,J＝2.7,1.8Hz,1H),6.19(d,J＝7.8Hz,1H),5.41(d,J＝8.0Hz,1H),5.01(q,J＝9.0Hz,2H),4.47(d,J＝6.1Hz,2H),3.62(s,3H),3.28–3.26(m,1H),2.36–2.18(m,7H),2.14–2.04(m,2H),1.91–1.81(m,2H),1.38–1.28(m,4H).
Compound 11-p2 spectral data: LC-MS (ESI) M/z 552.0 (M + H) + 。 1 H NMR(400MHz,DMSO-d 6 )δ8.36(t,J＝6.1Hz,1H),7.44(t,J＝7.6Hz,1H),7.40–7.30(m,3H),7.27(t,J＝2.0Hz,1H),7.01–6.92(m,2H),6.85(d,J＝8.3Hz,1H),6.69(t,J＝2.5Hz,1H),6.47(dd,J＝2.7,1.8Hz,1H),6.20(d,J＝7.8Hz,1H),5.32(d,J＝7.2Hz,1H),5.02(q,J＝9.0Hz,2H),4.47(d,J＝6.0Hz,2H),3.62(s,3H),3.59–3.54(m,1H),2.30–2.20(m,7H),1.86–1.76(m,4H),1.69–1.60(m,2H),1.58–1.50(m,2H).
Example 12: synthetic route to compound 12
Step 1: synthesis of Compound 12-1
To the reaction flask was added 2-fluoro-4-methylsulphonylaniline (500mg, 2.64mmol), DMF (10 mL), pinacol ester of 3-bromomethylphenyl) boronic acid (1.57g, 5.29mmol), cesium carbonate (2.58g, 7.93mmol). The mixture was stirred at 100 ℃ overnight. After the reaction was completed, the reaction was cooled to room temperature, and then ethyl acetate and water were added to dilute the reaction solution. The organic phase was separated and the aqueous phase was extracted with ethyl acetate. The combined organic phases were washed with saturated aqueous sodium chloride solution and concentrated under reduced pressure, and the crude product was purified by automatic column chromatography (Biotage) (mobile phase: petroleum ether/ethyl acetate 100/0 to 80/20) to give the title compound 12-1 as a brown oil (400mg, 37%). LC-MS (ESI): M/z 428.1 (M + Na) + .
And 2, step: synthesis of Compound 12
Add intermediate B (170mg, 0.365mmol), intermediate 12-1 (193mg, 0.475mmol), DMF (5 mL), water (1 mL), potassium carbonate (101mg, 0.730mmol) to the reaction flask, replace 3 to 5 times with nitrogen, add Pd (PPh) 3 ) 4 (42mg, 0.036mmol), and the nitrogen was further replaced 3 to 5 times. Mixture ofStirring for 2 hours at 100 ℃ under the protection of nitrogen, and then cooling to room temperature. After addition of saturated aqueous EDTA and stirring at room temperature for a further 1 hour, dilution is carried out with ethyl acetate and water. The organic phase was separated and the aqueous phase was extracted with ethyl acetate. Mixing organic phases, washing with saturated sodium chloride aqueous solution, drying with anhydrous sodium sulfate, filtering, concentrating under reduced pressure to obtain crude product of compound 12, and purifying compound 12
The preparation method comprises the following steps: a water (10 mmol NH) 4 HCO 3 ) And B, acetonitrile, wherein the preparation column comprises: welch Xtimate C18, 21.2 × 250mm,10um, gradient: 10% of B phase 2.5min, after 10min the B phase increased from 34% to 44%, retention was 44% B3 min, and 95% B retention was 6min, to give the target compound 12-p1 (15mg, 7%) as a white solid and 12-p2 (20mg, 9%) as a white solid.
12-p1 spectrum data: LC-MS (ESI) M/z 617.0 (M + H) + 。 1 H NMR(400MHz,DMSO-d 6 )δ7.51(dd,J＝11.4,2.1Hz,1H),7.49–7.43(m,2H),7.43–7.34(m,3H),7.32(td,J＝6.3,2.4Hz,1H),6.97(t,J＝7.9Hz,1H),6.83(d,J＝9.6Hz,2H),6.76(t,J＝8.5Hz,1H),6.19(d,J＝7.8Hz,1H),5.38(d,J＝8.0Hz,1H),4.97(q,J＝9.1Hz,2H),4.53(d,J＝6.2Hz,2H),3.30–3.27(m,1H),3.07(s,3H),2.23–2.17(m,7H),2.12–2.05(m,2H),1.88–1.81(m,2H),1.39–1.27(m,4H).
12-p2 spectrum data: LC-MS (ESI) M/z 617.0 (M + H) + 。 1 H NMR(400MHz,DMSO-d 6 )δ7.52(dd,J＝11.5,2.1Hz,1H),7.49–7.44(m,2H),7.44–7.35(m,3H),7.32(td,J＝6.3,2.4Hz,1H),7.02–6.94(m,2H),6.87(d,J＝8.2Hz,1H),6.76(t,J＝8.5Hz,1H),6.21(d,J＝7.7Hz,1H),5.28(d,J＝7.0Hz,1H),4.99(q,J＝9.0Hz,2H),4.53(d,J＝6.2Hz,2H),3.66–3.57(m,1H),3.07(s,3H),2.48–2.20(m,7H),1.91–1.76(m,4H),1.69–1.57(m,4H).
Example 13: synthetic route to compound 13
Step 1: synthesis of Compound 13-1
Adding into a reaction flask6-bromopyridylaldehyde (924mg, 4.97mmol), 2-methoxy-4- (methylsulfonyl) aniline (500mg, 2.48mmol) and methanol (10 mL). After stirring at room temperature for 30 minutes, 1.8mL of acetic acid was added to the reaction solution, followed by addition of sodium cyanoborohydride (186mg, 3.00mmol). Stirred at room temperature for 5 hours. The reaction was quenched with saturated ammonium chloride and diluted with ethyl acetate and water. The organic phase was separated and the aqueous phase was extracted with ethyl acetate. Combining the organic phases, washing with water, washing with saturated NaCl, anhydrous Na 2 SO 4 Drying, filtration, evaporation to dryness and purification by column (mobile phase: petroleum ether/ethyl acetate 3/1 to 2/1) gave compound 13-1 (950mg, 95%). LC-MS (ESI) m/z371.0;373.0 (M + H) + .
Step 2: synthesis of Compound 13-2
Add intermediate B (500mg, 1.08mmol), pinacolborane (963mg, 7.53mmol), potassium acetate (316mg, 3.23mmol), S-Phos (88mg, 0.22mmol), pd to the reaction flask 2 (dba) 3 (98mg, 0.11mmol), toluene (20 mL). The mixture was purged with nitrogen and stirred at 60 ℃ for 3 hours. The reaction solution was cooled to room temperature and diluted with ethyl acetate and water. The organic phase was separated and the aqueous phase was extracted with ethyl acetate. Combining the organic phases, washing with water, washing with saturated NaCl, anhydrous Na 2 SO 4 Dried, filtered, evaporated to dryness and purified by column chromatography (mobile phase: dichloromethane/methanol 15/1 to 10/1) to give compound 13-2 (370mg, 74%). LC-MS (ESI): M/z 466.3 (M + H) + .
And 3, step 3: synthesis of Compound 13
13-1 (170mg, 0.37mmol), 13-2 (136mg, 0.37mmol), potassium carbonate (101mg, 0.74mmol), pd (PPh) were added to the reaction flask 3 ) 4 (43mg, 0.04mmol), N, N-dimethylformamide (5 mL) and water (1 mL). N for reaction solution 2 The mixture was replaced and stirred at 100 ℃ for 2 hours. Dilute with ethyl acetate and water. The organic phase was separated and the aqueous phase was extracted with ethyl acetate. Combining the organic phases, washing with water, washing with saturated NaCl, anhydrous Na 2 SO 4 Drying, filtering, and steamingDry to give crude compound 13, compound 13 was purified by silica gel column) (mobile phase: dichloromethane/methanol 12/1 to 10/1) to give compound 13-p1 (51 mg) and compound 13-p2 (74 mg) as white solids.
Compound 13-p1 spectral data: LC-MS (ESI) M/z 630.3 (M + H) +; 1 H NMR(400MHz,DMSO-d 6 ):δ7.83(1H,t,J＝7.8Hz),7.67(1H,d,J＝7.9Hz),7.44(1H,s),7.28–7.20(3H,m),7.03(1H,t,J＝8.0Hz),6.84(1H,d,J＝8.3Hz),6.75(1H,t,J＝6.2Hz),6.52(1H,d,J＝8.9Hz),6.19(1H,d,J＝7.9Hz),5.84(2H,d,J＝8.1Hz),5.52(1H,d,J＝8.0Hz),4.56(2H,d,J＝6.2Hz),3.94(3H,s),3.06(3H,s),2.20(6H,s),2.11(2H,d,J＝11.5Hz),1.85(2H,d,J＝11.6Hz),1.32(6H,dd,J＝18.3,10.2Hz).
compound 13-p2 spectral data: LC-MS (ESI) M/z 630.4 (M + H) + ； 1 H NMR(400MHz,DMSO-d 6 ):δ7.83(1H,t,J＝7.8Hz),7.70(1H,d,J＝7.9Hz),7.57(1H,s),7.23(3H,dd,J＝5.2,3.0Hz),7.03(1H,t,J＝8.0Hz),6.85(1H,d,J＝8.4Hz),6.75(1H,t,J＝6.3Hz),6.53(1H,d,J＝8.9Hz),6.20(1H,d,J＝7.8Hz),5.85(2H,d,J＝9.4Hz),5.44(1H,d,J＝7.5Hz),4.56(2H,d,J＝6.2Hz),3.94(3H,s),3.56(1H,s),3.06(3H,s),2.19(6H,s,6H),2.07(1H,s),1.89–1.73(4H,m),1.67(2H,dd,J＝9.1,4.2Hz,2H),1.59–1.46(2H,m).
Example 14: synthetic route to Compound 14
Step 1: synthesis of Compound 14-1
To the reaction flask were added intermediate B-2 (200mg, 0.59mmol), 1, 4-cyclohexanedione monoethylene ketal (470mg, 3mmol), stannous chloride (27mg, 0.14mmol), polymethylhydrosiloxane (268mg, 1.2mmol), and methanol (20 mL). The mixture was stirred for 2 hours at 70 ℃ under nitrogen. Add ethyl acetate and water to dilute. The organic phase was separated and the aqueous phase was extracted with ethyl acetate. Mixing organic phases, washing with saturated sodium chloride aqueous solution, concentrating under reduced pressure, and purifying the crude product with automatic column chromatography (Biotage) (mobile phase: petroleum ether/ethyl acetate 100/0-60/40) to obtain yellow solidCompound 14-1 (302mg, 93%) was targeted. LC-MS (ESI) M/z 481.1 (M + H) + .
And 2, step: synthesis of Compound 14-2
Add intermediate C (152mg, 0.31mmol), intermediate 14-1 (198mg, 0.47mmol), palladium tetrakistriphenylphosphine (37mg, 0.03mmol), potassium carbonate (87mg, 0.63mmol), N, N-dimethylformamide (5 mL), water (1 mL) to the reaction flask. The nitrogen was replaced 3 to 5 times. The mixture was stirred at 100 ℃ for 2 hours under nitrogen. After the reaction, ethyl acetate and brine were added to dilute the reaction mixture. The organic phase was separated and the aqueous phase was extracted with ethyl acetate. The combined organic phases were washed with saturated aqueous sodium chloride solution and concentrated under reduced pressure, and the crude product was purified by automatic column chromatography (Biotage) (mobile phase: petroleum ether/ethyl acetate 100/0 to 20/80) to give intermediate 14-2 (130mg, 65%) as a brown solid. LC-MS (ESI) M/z 644.1 (M + H) + .
And 3, step 3: synthesis of Compound 14-3
Intermediate 14-2 (130mg, 0.2mmol), 4N diluted hydrochloric acid (4 mL), tetrahydrofuran (2 mL), trifluoroacetic acid (3 mL) was added to the reaction flask. Stirred at 50 ℃ overnight. The next day, tetrahydrofuran and trifluoroacetic acid were distilled off. The pH was adjusted to 8-9 with saturated aqueous sodium bicarbonate. Ethyl acetate and water. The organic phase was separated and the aqueous phase was extracted with ethyl acetate. The combined organic phases were washed with saturated sodium chloride, dried over anhydrous sodium sulfate, filtered and evaporated to dryness to give compound 14-3 (65mg, 54%). LC-MS (ESI): M/z 600.1 (M + H) + .
And 4, step 4: synthesis of Compound 14
Adding 2-oxazole-7-azaspiro [3.5 ] into a reaction bottle]Nonane hemioxalate (110mg, 0.32mmol), triethylamine (70mg, 0.69mmol), methanol (3 mL) were stirred at 25 ℃ for 0.5h. Compound 14-3 (65mg, 0.11mmol), tetraisopropyl titanate (4 mL) was added. Reaction solution is introduced with N 2 Balloon, stir overnight at 55 ℃. The next day, methanol (2 mL), sodium borohydride acetate (70mg, 0.33mmol) was added and stirred at 25 ℃ for 3h. The reaction solution was quenched with saturated aqueous sodium bicarbonate. Ethyl acetate and water. The organic phase was separated and the aqueous phase was extracted with ethyl acetate. Mixing organic phases, washing with saturated sodium chloride, drying with anhydrous sodium sulfate, filtering, evaporating to obtain crude productAnd (4) an object 14. Preparation and purification of Compound 14 (Welch XTimate C18, 21.2X 250mm,10um, water (10 mM ammonium bicarbonate)/acetonitrile, flow rate 30mL/min, column temperature 25 ℃, detection wavelength 254 nm) gave the target compound 14-p1 (1mg, 1%), 14-p2 (2mg, 2%) as a white solid.
14-p1 spectral data: LC-MS (ESI) M/z 711.1 (M + H) +. 1 H NMR (400 MHz, deuterated chloroform) delta 7.47 (1H, t, J = 7.5Hz), 7.42-7.34 (3H, m), 7.30-7.27 (1H, m), 7.24 (1H, d, J = 2.0Hz), 7.15 (1H, t, J = 8.0Hz), 6.82 (1H, d, J = 8.3Hz), 6.54 (1H, d, J = 8.4Hz), 6.43 (1H, d, J = 0.8Hz), 6.33 (1H, d, J = 7.7Hz), 5.40-5.29 (2H, m), 4.60-4.41 (7H, m), 3.93 (3H, s), 3.53-3.38 (3H, m), 3.09 (1H, s), 3.01 (3H, s), 2.72-2.61 (3H, m), 2.41 (3H, d, J = 11.9Hz), 2.33-2.24 (2H, m), 2.08-1.93 (2H, m), 1.76 (2H, d, J = 12.1Hz), 1.34-1.31 (3H, m).
14-p2 spectral data: LC-MS (ESI) M/z 711.1 (M + H) +. 1 H NMR (400 MHz, deuterated chloroform) delta 7.47 (1H, t, J = 7.5Hz), 7.42-7.34 (3H, m), 7.30-7.27 (1H, m), 7.24 (1H, d, J = 2.0Hz), 7.15 (1H, t, J = 8.0Hz), 6.82 (1H, d, J = 8.3Hz), 6.54 (1H, d, J = 8.4Hz), 6.43 (1H, d, J = 0.8Hz), 6.33 (1H, d, J = 7.7Hz), 5.40-5.29 (2H, m), 4.60-4.41 (7H, m), 3.93 (3H, s), 3.53-3.38 (3H, m), 3.09 (1H, s), 3.01 (3H, s), 2.72-2.61 (3H, m), 2.41 (3H, d, J = 11.9Hz), 2.33-2.24 (2H, m), 2.08-1.93 (2H, m), 1.76 (2H, d, J = 12.1Hz), 1.34-1.31 (3H, m).
Example 15: synthetic route to Compound 15
Step 1: synthesis of Compound 15-1
To the reaction flask was added (3-bromomethylphenyl) boronic acid pinacol ester (400mg, 1.35mmol), DMF (10 mL), (2-aminobenzyl) -carbamic acid tert-butyl ester (449mg, 2.02mmol), potassium carbonate (558mg, 4.04mmol). The mixture was stirred at 70 ℃ for 3 hours. After the reaction, ethyl acetate and water were added to dilute. The organic phase was separated and the aqueous phase was extracted with ethyl acetate. The organic phases are combined, washed with saturated aqueous sodium chloride solution, concentrated under reduced pressure and the crude product is purified by means of an automatic column chromatography (Biotage) (mobile phase: petroleum ether)Ethyl acetate 100/0 to 88/12) to give the title compound 15-1 (387mg, 66%) as a yellow solid. LC-MS (ESI): M/z 439.0 (M + H) + .
Step 2: synthesis of Compound 15-2
To a reaction flask were added 15-1 (150mg, 0.322mmol), compound B (198mg, 0.451mmol), DMF (5 mL), water (1 mL), potassium carbonate (89mg, 0.644mmol), nitrogen-exchanged 3 to 5 times, and Pd (PPh) was added 3 ) 4 (37mg, 0.032mmol), and then replaced with nitrogen 3 to 5 times. The mixture was stirred at 100 ℃ for 2 hours under nitrogen and then cooled to room temperature. After addition of saturated aqueous EDTA and stirring at room temperature for a further 1 hour, dilution is carried out with ethyl acetate and water. The organic phase was separated and the aqueous phase was extracted with ethyl acetate. The combined organic phases were washed with saturated aqueous sodium chloride and concentrated under reduced pressure, and the crude product was purified by an automatic column chromatography (Biotage) (mobile phase: dichloromethane/0.5M ammonia in methanol 100/0 to 90/10) to give target compound 15-2 (186mg, 89%) as an orange solid.
And step 3: synthesis of Compound 15
Add intermediate 15-2 (180mg, 0.277mmol), dichloromethane (8 mL), trifluoroacetic acid (4 mL) to the reaction flask. The mixture was stirred at room temperature for 1 hour. After the reaction, the reaction mixture was slowly poured into a saturated aqueous sodium bicarbonate solution, and ethyl acetate was added. The organic phase was separated and the aqueous phase was extracted with ethyl acetate. The organic phases were combined, washed with saturated aqueous sodium chloride, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to give crude compound 15, which was purified by preparative purification (Boston pHlex ODS, 21.2X 250mm,10um, water (0.05% trifluoroacetic acid)/acetonitrile, flow rate 30mL/min, column temperature 25 ℃, detection wavelength 254 nm) to give target compound 15-p1 (5mg, 3%) as a white solid and target compound 15-p2 (15mg, 10%) as a white solid.
Compound 15-p1 spectral data: LC-MS (ESI) M/z 550.3 (M + H) + 。 1 H NMR(400MHz,DMSO-d 6 )δ7.50–7.40(m,3H),7.34(dt,J＝6.7,2.1Hz,1H),7.04(dd,J＝7.4,1.6Hz,1H),7.00–6.92(m,2H),6.84(s,1H),6.81(d,J＝8.3Hz,1H),6.66(t,J＝6.1Hz,1H),6.51(t,J＝7.3Hz,1H),6.47(d,J＝8.0Hz,1H),6.19(d,J＝7.8Hz,1H),5.36(d,J＝8.0Hz,1H),4.95(q,J＝9.0Hz,2H),4.43(d,J＝5.2Hz,2H),3.76(s,2H),3.28–3.27(m,1H),2.19–2.12(m,7H),2.11–2.03(m,2H),1.88–1.79(m,2H),1.39–1.24(m,4H).
Compound 15-p2 spectral data: LC-MS (ESI) M/z 550.3 (M + H) + 。 1 H NMR(400MHz,DMSO-d 6 )δ7.49–7.40(m,3H),7.35(dt,J＝6.7,2.1Hz,1H),7.04(dd,J＝7.4,1.6Hz,1H),7.00–6.92(m,3H),6.82(d,J＝8.2Hz,1H),6.72–6.60(m,1H),6.52(td,J＝7.4,1.1Hz,1H),6.47(d,J＝8.0Hz,1H),6.20(d,J＝7.8Hz,1H),5.30(d,J＝7.5Hz,1H),4.95(q,J＝9.0Hz,2H),4.43(d,J＝4.8Hz,2H),3.77(s,2H),3.59–3.47(m,1H),2.16(s,6H),2.07–2.02(m,1H),1.85–1.71(m,4H),1.69–1.57(m,2H),1.54–1.44(m,2H).
Example 16: synthetic route to compound 16
Step 1 Synthesis of Compound 16-1
To the reaction flask was added N- (4-amino-3-methoxyphenyl) methanesulfonamide (500mg, 2.48mmol), 5-bromothiophene-2-carbaldehyde (949mg, 4.97mmol), methanol (20 mL), acetic acid (1.85 mL), naBH 3 CN (187mg, 2.98mmol). The mixture was stirred at 50 ℃ overnight. After the reaction, saturated aqueous sodium bicarbonate was added and stirred for 15 minutes, followed by addition of ethyl acetate. The organic phase was separated and the aqueous phase was extracted with ethyl acetate. The combined organic phases were washed with saturated aqueous sodium chloride solution and concentrated under reduced pressure, and the crude product was purified by automatic column chromatography (Biotage) (mobile phase: petroleum ether/ethyl acetate 100/0 to 55/45) to give the title compound 16-1 (410mg, 44%) as a yellow solid. LC-MS (ESI) M/z 375.8 (M + H) + .
Step 2: synthesis of Compound 16
Add intermediate 13-2 (120mg, 0.318mmol), intermediate 16-1 (178mg, 0.382mmol), DMF (5 mL), water (1 mL), potassium carbonate (88mg, 0.637mmol) to the reaction flask, replace 3 to 5 times with nitrogen, and add Pd (PPh) 3 ) 4 (37mg, 0.032mmol), and then replaced with nitrogen 3 to 5 times. The mixture was stirred at 100 ℃ for 2 hours under nitrogen and then cooled to room temperature. Adding intoAfter saturating the aqueous EDTA solution and stirring at room temperature for 1 hour, add ethyl acetate and water to dilute. The organic phase was separated and the aqueous phase was extracted with ethyl acetate. The organic phases were combined, washed with saturated aqueous sodium chloride, concentrated under reduced pressure, and the resulting crude product was purified by automatic column chromatography (Biotage) (mobile phase: dichloromethane/0.5M ammonia in methanol 100/0 to 90/10) to give compound 16, which was further subjected to preparative purification (Boston pHlex ODS, 21.2X 250mm,10um, water (0.05% trifluoroacetic acid)/acetonitrile, flow rate of 30mL/min, column temperature 25 ℃ C., detection wavelength 254 nm) to give 16-p1 (47mg, 23%) as a white solid target compound and 16-p2 (75mg, 37%) as a white solid target compound.
Compound 16-p1 spectral data: LC-MS (ESI) M/z 635.0 (M + H) + 。 1 H NMR(400MHz,DMSO-d 6 )δ7.31(dd,J＝8.4,2.0Hz,1H),7.23(d,J＝2.0Hz,1H),7.15–7.08(m,2H),7.00–6.93(m,2H),6.79–6.70(m,3H),6.16(d,J＝7.8Hz,1H),5.46(d,J＝8.0Hz,1H),5.08(q,J＝9.0Hz,2H),4.64(d,J＝6.2Hz,2H),3.91(s,3H),3.29–3.23(m,1H),3.09(s,3H),2.18(s,7H),2.09–2.02(m,2H),1.86–1.77(m,2H),1.30–1.21(m,4H).
Compound 16-p2 spectral data: LC-MS (ESI): M/z 635.0 (M + H) + 。 1 H NMR(400MHz,DMSO-d 6 )δ7.31(dd,J＝8.4,2.0Hz,1H),7.23(d,J＝2.0Hz,1H),7.15–7.10(m,2H),7.07(s,1H),6.97(t,J＝8.0Hz,1H),6.81–6.69(m,3H),6.17(d,J＝7.9Hz,1H),5.40(d,J＝7.4Hz,1H),5.08(q,J＝9.0Hz,2H),4.64(d,J＝6.2Hz,2H),3.91(s,3H),3.60–3.53(m,1H),3.09(s,3H),2.18(s,6H),2.12–2.05(m,1H),1.85–1.70(m,4H),1.66–1.57(m,2H),1.53–1.44(m,2H).
Example 17: synthetic route to Compound 17
Step 1: synthesis of Compound 17-1
Into a reaction flask were charged N- (4-amino-3-methoxyphenyl) methanesulfonamide (322mg, 1.6 mmol), 4-bromomethyl-3-fluorophenylboronic acid pinacol ester (1g, 3.2mmol), cesium carbonate (1.6 g, 4.8mmol),n, N-dimethylformamide (15 mL). The mixture was stirred overnight at 100 ℃ under nitrogen. Add ethyl acetate and water to dilute. The organic phase was separated and the aqueous phase was extracted with ethyl acetate. The combined organic phases were washed with saturated aqueous sodium chloride and concentrated under reduced pressure, and the crude product was purified by autosampler (Biotage) (mobile phase: dichloromethane/methanol 100/0 to 70/30) to give the title compound 17-1 as a yellow solid (175mg, 25%). LC-MS (ESI): M/z 436.1 (M + H) + .
And 2, step: synthesis of Compound 17
To a reaction flask, compound 17-1 (175mg, 0.4mmol), intermediate B (130mg, 0.28mol), palladium tetrakistriphenylphosphine (35mg, 0.03mmol), potassium carbonate (78mg, 0.56mmol), N, N-dimethylformamide (5 mL), and water (1 mL) were added. The nitrogen was purged 3 to 5 times. The mixture was stirred at 100 ℃ for 1.5 hours under nitrogen. After the reaction, ethyl acetate and brine were added to dilute the reaction mixture. The organic phase was separated and the aqueous phase was extracted with ethyl acetate. The combined organic phases were washed with saturated aqueous sodium chloride solution and concentrated under reduced pressure, and the crude product was purified by automatic column chromatography (Biotage) (mobile phase: dichloromethane/methanol 100/0 to 80/20) to give compound 17 as a brown solid, and compound 17 was purified by preparative chromatography (Welch Xtimate C18, 21.2X 250mm,10um, water (10 mM ammonium bicarbonate)/acetonitrile, flow rate 30mL/min, column temperature 25 ℃ C., detection wavelength 254 nm) to give target compound 17-p1 (21mg, 12%) and 17-p2 (6mg, 3%) as white solids.
17-p1 spectrum data: LC-MS (ESI): M/z 647.1 (M + H) + 。 1 H NMR(400MHz,DMSO-d 6 )δ7.42–7.34(2H,m),7.29(2H,td,J＝8.0,1.8Hz),7.23(1H,d,J＝2.0Hz),7.06–6.96(2H,m),6.92(1H,d,J＝8.2Hz),6.64(1H,t,J＝6.4Hz),6.56(1H,d,J＝8.5Hz),6.22(1H,d,J＝7.8Hz),5.21(1H,d,J＝6.3Hz),5.07(2H,q,J＝9.0Hz),4.54(2H,d,J＝6.4Hz),3.93(3H,s),3.75–3.70(1H,m),3.08(3H,s),2.67(6H,s),2.51(1H,s),2.00(2H,d,J＝14.2Hz),1.79(4H,d,J＝12.5Hz),1.65(2H,d,J＝13.4Hz).
17-p2 spectrum data: LC-MS (ESI): M/z 647.1 (M + H) + 。 1 H NMR(400MHz,DMSO-d 6 )δ7.42–7.33(2H,m),7.33–7.25(2H,m),7.24(1H,d,J＝2.0Hz),6.98(1H,t,J＝8.0Hz),6.90(1H,s),6.84(1H,d,J＝8.3Hz),6.65–6.55(2H,m),6.20(1H,d,J＝7.8Hz),5.37(1H,d,J＝8.0Hz),5.05(2H,q,J＝9.0Hz),4.54(2H,d,J＝6.3Hz),3.93(3H,s),3.43(1H,s),3.08(3H,s),2.52(1H,s),2.19(6H,s),2.08(2H,d,J＝11.6Hz),2.03–1.94(1H,m),1.84(2H,d,J＝11.8Hz),1.33(3H,dd,J＝23.8,10.8Hz).
EXAMPLE 18 synthetic route to Compound 18
Step 1: synthesis of Compound 18-1
To the reaction flask were added N- (4-amino-3-methoxyphenyl) methanesulfonamide (261mg, 1.3mmol), 3-bromomethyl-4-fluorobenzeneboronic acid pinacol ester (0.8g, 2.5mmol), cesium carbonate (1.3g, 4mmol), and N, N-dimethylformamide (15 mL). The mixture was stirred overnight at 100 ℃ under nitrogen. Add ethyl acetate and water to dilute. The organic phase was separated and the aqueous phase was extracted with ethyl acetate. The organic phases were combined, washed with saturated aqueous sodium chloride solution, concentrated under reduced pressure and the resulting crude product was purified by an automatic column chromatography (Biotage) (mobile phase: dichloromethane/methanol 100/0 to 70/30) to give target compound 18-1 (192 mg, crude product) as a yellow solid and boric acid (116 mg, crude product) as a hydrolysate of 18-1. LC-MS (ESI): M/z 436.1 (M + H) + .
And 2, step: synthesis of Compound 18
Add 18-1 and its hydrolyzate (ca. 200mg, crude), and palladium tetrakistriphenylphosphine (35mg, 0.03mmol), potassium carbonate (78mg, 0.56mmol), N, N-dimethylformamide (5 mL), water (1 mL) to the reaction flask. The nitrogen was replaced 3 to 5 times. The mixture was stirred at 100 ℃ for 1 hour under nitrogen. After the reaction, ethyl acetate and brine were added to dilute the reaction mixture. The organic phase was separated and the aqueous phase was extracted with ethyl acetate. The combined organic phases were washed with saturated aqueous sodium chloride solution and concentrated under reduced pressure, and the crude product was purified by automatic column chromatography (Biotage) (mobile phase: dichloromethane/methanol 100/0 to 80/20) to give the desired product 18 as a brown solid. Purification of the solid obtained from Compound 18 (Welch Xtimate C18, 21.2X 250mm,10um, water (10 mM ammonium bicarbonate)/acetonitrile, flow rate 30mL/min, column temperature 25 ℃, detection wavelength 254 nm) gave the target compound 18-p1 (35mg, 30%), 18-p2 (13mg, 11%) as a white solid.
Compound 18-p1 spectral data: LC-MS (ESI) M/z 647.1 (M + H) + 。 1 H NMR(400MHz,DMSO-d 6 )δ7.43–7.34(3H,m),7.25(1H,dd,J＝8.4,2.0Hz),7.20(1H,d,J＝2.0Hz),6.97(1H,dd,J＝6.8,2.7Hz),6.89(1H,s),6.84(1H,d,J＝10.5Hz),6.59–6.54(1H,m),6.22–6.17(1H,m),5.25(2H,dd,J＝25.6,8.1Hz),4.87(2H,q,J＝9.0Hz),4.55(2H,d,J＝6.3Hz),3.89(3H,s),3.56(1H,dd,J＝5.8,2.8Hz),3.04(3H,s),2.55(1H,dd,J＝5.3,1.3Hz),2.25(6H,s),1.81(4H,dt,J＝11.3,5.1Hz),1.68–1.62(2H,m),1.57–1.50(2H,m).
Compound 18-p2 spectral data: LC-MS (ESI): M/z 647.1 (M + H) + 。 1 H NMR(400MHz,DMSO-d 6 )δ7.43–7.34(3H,m),7.25(1H,dd,J＝8.3,2.0Hz),7.20(1H,d,J＝2.0Hz),6.95(1H,d,J＝8.0Hz),6.84(1H,s),6.79(1H,d,J＝5.8Hz),6.60(1H,t,J＝6.3Hz),6.55(1H,d,J＝8.4Hz),6.21–6.16(1H,m),5.37(1H,d,J＝7.5Hz),4.87(2H,q,J＝8.8Hz),4.54(2H,d,J＝6.4Hz),3.89(3H,s),3.71–3.61(1H,m),3.04(3H,s),2.34–2.30(1H,m),2.26(6H,s),2.12–2.02(3H,m),1.91–1.80(3H,m),1.34(2H,d,J＝8.5Hz).
Example 19: synthetic route to compound 19
Step 1: synthesis of Compound 19-1
To the reaction flask were added N- (4-amino-3-methoxyphenyl) methanesulfonamide (312mg, 1.5 mmol), 3-bromomethyl-2-fluorophenylboronic acid pinacol ester (2.4g, 7.6 mmol), cesium carbonate (1g, 3mmol), and N, N-dimethylformamide (20 mL). The mixture was stirred overnight at 70 ℃ under nitrogen. Add ethyl acetate and water to dilute. The organic phase was separated and the aqueous phase was extracted with ethyl acetate. The combined organic phases were washed with saturated aqueous sodium chloride solution and concentrated under reduced pressure, and the resulting crude product was purified by an automatic column chromatography (Biotage) (mobile phase: dichloromethane/methanol 100/0 to 65/35) to give the title compound 19-1 (636 mg, crude) as a yellow solid. LC-MS (ESI) M/z 436.1 (M + H) + .
And 2, step: synthesis of Compound 19
Add 19-1 (210 mg, crude), intermediate B (160mg, 0.34mol), tetrakistriphenylphosphine palladium (40mg, 0.03mmol), potassium carbonate (90mg, 0.65mmol), N, N-dimethylformamide (5 mL), water (1 mL) to the reaction flask. The nitrogen was purged 3 to 5 times. The mixture was stirred at 100 ℃ for 1 hour under nitrogen. After the reaction, ethyl acetate and brine were added to dilute the reaction mixture. The organic phase was separated and the aqueous phase was extracted with ethyl acetate. The combined organic phases were washed with saturated aqueous sodium chloride solution, concentrated under reduced pressure and the crude product purified by column autosampler (Biotage) (mobile phase: dichloromethane/methanol 100/0 to 80/20) to give compound 19 as a brown solid. Compound 19 was purified by preparative purification (Welch Xtimate C18, 21.2X 250mm,10um, water (10 mM ammonium bicarbonate)/acetonitrile, flow rate 30mL/min, column temperature 25 ℃, detection wavelength 254 nm) to give the title compound 19-p1 (6 mg, 3%), 19-p2 (20mg, 9%) as a white solid.
Compound 19-p1 spectral data: LC-MS (ESI) M/z 647.1 (M + H) + 。1H NMR(400MHz,DMSO-d 6 )δ7.41–7.34(2H,m),7.31–7.19(3H,m),7.05–6.96(2H,m),6.88(1H,d,J＝8.2Hz),6.66(1H,t,J＝6.4Hz),6.56(1H,d,J＝8.4Hz),6.22(1H,d,J＝7.8Hz),5.34(1H,d,J＝7.2Hz),4.92(2H,q,J＝9.1Hz),4.55(2H,d,J＝6.4Hz),3.93(3H,s),3.65–3.57(1H,m),3.08(3H,s),2.51(1H,s),2.39–2.16(6H,m),1.85(4H,q,J＝15.3,11.6Hz),1.70–1.55(4H,m).
Compound 19-p2 spectral data: LC-MS (ESI) M/z 647.1 (M + H) + 。1H NMR(400MHz,DMSO-d 6 )δ7.37(2H,q,J＝7.3Hz),7.31–7.19(3H,m),7.01(1H,t,J＝8.0Hz),6.91(1H,s),6.85(1H,d,J＝8.2Hz),6.65(1H,t,J＝6.4Hz),6.56(1H,d,J＝8.4Hz),6.21(1H,d,J＝7.9Hz),5.43(1H,d,J＝8.1Hz),4.92(2H,q,J＝9.0Hz),4.55(2H,d,J＝6.3Hz),3.93(3H,s),3.25(1H,d,J＝9.3Hz),3.08(3H,s),2.51(1H,s),2.20(6H,s),2.09(2H,d,J＝12.2Hz),1.85(2H,d,J＝11.9Hz),1.37–1.25(4H,m).
Example 20: synthetic route to compound 20
Step 1: synthesis of Compound 15-1
To the reaction flask was added (3-bromomethylphenyl) boronic acid pinacol ester (200mg, 0.673mmol), DMF (10 mL), (2-aminobenzyl) -carbamic acid tert-butyl ester (224mg, 1.01mmol), potassium carbonate (279mg, 2.02mmol). The mixture was stirred at 70 ℃ for 4 hours. After the reaction, ethyl acetate and water were added to dilute the reaction mixture. The organic phase was separated and the aqueous phase was extracted with ethyl acetate. The combined organic phases were washed with saturated aqueous sodium chloride solution and concentrated under reduced pressure, and the crude product was purified by an automatic column chromatography (Biotage) (mobile phase: petroleum ether/ethyl acetate 100/0 to 85/15) to give the desired compound 15-1 (189mg, 64%) as a yellow solid. LC-MS (ESI) M/z 439.0 (M + H) + .
Step 2: synthesis of Compound 20
Add 15-1 (140mg, 0.300mmol), intermediate B (158mg, 0.361mmol), DMF (5 mL), water (1 mL), potassium carbonate (83mg, 0.601mmol), nitrogen exchange 3-5 times, add Pd (PPh) 3 ) 4 (35mg, 0.030mmol), nitrogen is replaced 3 to 5 times. The mixture was stirred at 100 ℃ for 2 hours under nitrogen and then cooled to room temperature. After addition of saturated aqueous EDTA and stirring at room temperature for a further 1 hour, it is diluted with ethyl acetate and water. The organic phase was separated and the aqueous phase was extracted with ethyl acetate. The combined organic phases were washed with saturated aqueous sodium chloride solution and concentrated under reduced pressure, and the crude product was purified by an automatic column chromatography (Biotage) (mobile phase: dichloromethane/0.5M ammonia in methanol 100/0 to 90/10) to give target compound 20 as an orange solid (141mg, 72%). LC-MS (ESI) M/z 650.5 (M + H) + .
EXAMPLE 21 Synthesis of Compound 21
Step 1: synthesis of Compound 21-1
To a solution of 1H-pyrazole-3-carboxylic acid (45.28mg, 0.404mmol) in DMF (3 mL) were added DIPEA (94.91mg, 0.734mmol) and HATU (209.44mg, 0.551mmol) and the reaction was stirred at room temperature for 0.5H. Then 2-methylpropan-2-yl ({ 2- [3- (aminomethyl) benzene)Base of]-1- (2, 2-trifluoroethyl) indol-4-yl } [4- (dimethylamino) cyclohexyl]Carbamate (200mg, 0.367mmol) was stirred at room temperature for 1 hour. The reaction solution was diluted with ethyl acetate and water. The organic phase was separated and the aqueous phase was extracted with ethyl acetate. The organic phases were combined, washed with water, saturated NaCl, anhydrous Na 2 SO 4 Drying, filtration, evaporation to dryness and purification on column (mobile phase: dichloromethane/0.5M amine methanol 0/1 to 10/1) gave compound 21-1 (120mg, 51.2%). LC-MS (ESI) M/z 639.6 (M + H) + .
And 2, step: synthesis of Compound 21
Add compound 21-1 (120mg, 0.43mmol), DCM (3 mL), TFA (0.6 mL, 7.835mmol) to the reaction flask and stir at room temperature for 1h. The reaction solution was spin-dried, and subjected to prep-HPLC to obtain compound 21 (32mg, 32%). LC-MS (ESI) M/z 539.3 (M + H) + ； 1 H NMR(400MHz,DMSO-d6)δ13.28(s,1H),8.82(s,1H),7.77(s,1H),7.50–7.41(m,2H),7.40–7.32(m,2H),7.02–6.93(m,2H),6.84(d,J＝8.3Hz,1H),6.68(s,1H),6.20(d,J＝7.8Hz,1H),5.34(d,J＝7.5Hz,1H),5.02(q,J＝9.0Hz,2H),4.51(d,J＝6.3Hz,2H),3.53(s,1H),2.18(s,6H),1.79(dd,J＝11.5,7.5Hz,4H),1.65(dq,J＝11.3,7.4,5.9Hz,2H),1.57–1.43(m,2H).
EXAMPLE 22 synthetic route to Compound 22
Step 1: synthesis of Compound 22-1
To a solution of 5-methyl-1H-pyrazole-3-carboxylic acid (50.94mg, 0.404mmol) in DMF (3 mL) were added DIPEA (94.91mg, 0.734mmol) and HATU (209.44mg, 0.551mmol), and the reaction was stirred at room temperature for 0.5H. Then 2-methylpropan-2-yl ({ 2- [3- (aminomethyl) phenyl)]-1- (2, 2-trifluoroethyl) indol-4-yl } [4- (dimethylamino) cyclohexyl]Carbamate (200mg, 0.367mmol) was stirred at room temperature for 1 hour. The reaction solution was diluted with ethyl acetate and water. The organic phase was separated and the aqueous phase was extracted with ethyl acetate. Mixing organic phases, washing with water, washing with saturated NaCl, drying with anhydrous Na2SO4, filtering, evaporating to dryness, purifying with columnThe resulting mixture was reacted (mobile phase: dichloromethane/0.5M amine methanol 0/1 to 10/1) to obtain compound 22-1 (85mg, 35.5%). LC-MS (ESI): M/z 653.6 (M + H) + .
Step 2: synthesis of Compound 22
To the reaction flask was added 2-methylpropan-2-yl ({ 2- [3- (aminomethyl) phenyl)]-1- (2, 2-trifluoroethyl) indol-4-yl } [4- (dimethylamino) cyclohexyl]Amino) formate (85mg, 0.13mmol), DCM (3 mL), TFA (0.6mL, 7.835mmol), was stirred at room temperature for 1 hour. The reaction mixture was spin-dried, and subjected to prep-HPLC to obtain Compound 23 (20mg, 28.6%). LC-MS (ESI) M/z553.3 (M + H) + .1H NMR(400MHz,DMSO-d6)δ12.92(s,1H),8.70(s,1H),7.43(dd,J＝15.0,7.5Hz,2H),7.35(t,J＝8.5Hz,2H),7.01–6.92(m,2H),6.84(d,J＝8.2Hz,1H),6.38(s,1H),6.20(d,J＝7.8Hz,1H),5.34(d,J＝7.5Hz,1H),5.03(q,J＝9.0Hz,2H),4.48(d,J＝6.3Hz,2H),3.52(d,J＝11.3Hz,1H),2.24(s,3H),2.18(s,6H),1.78(d,J＝9.8Hz,4H),1.64(dd,J＝8.7,4.2Hz,2H),1.51(d,J＝13.1Hz,3H).
EXAMPLE 23 synthetic route to Compound 23
Step 1: synthesis of Compound 23-1
To the reaction flask were added (3-bromomethylphenyl) boronic acid pinacol ester (1g, 3.37mmol), DMF (20 mL), 2-aminobenzamide (917mg, 6.73mmol), potassium carbonate (1.4g, 10.10mmol). The mixture was stirred at 70 ℃ for 3 hours. After the reaction, ethyl acetate and water were added to dilute the reaction mixture. The organic phase was separated and the aqueous phase was extracted with ethyl acetate. The combined organic phases were washed with saturated aqueous NaCl solution and concentrated under reduced pressure, and the crude product was purified by automatic column chromatography (Biotage) (mobile phase: petroleum ether/ethyl acetate 100/0 to 40/60) to give the title compound 23-1 as a white solid (902mg, 76%). LC-MS (ESI) M/z 353.1 (M + H) + .
Step 2: synthesis of Compound 23
Add intermediate B (50mg, 0.107mmol), compound 23-1 (57mg, 0.161mmol), DMF (3 mL), water (0.6 mL), potassium carbonate (30mg, 0.214mmol)) Replacing 3-5 times with nitrogen, and adding Pd (PPh) 3 ) 4 (12mg, 0.010mmol), and the nitrogen gas was further replaced 3 to 5 times. The mixture was stirred at 100 ℃ for 2 hours under nitrogen and then cooled to room temperature. Add ethyl acetate and water to dilute. The organic phase was separated and the aqueous phase was extracted with ethyl acetate. The combined organic phases were washed with saturated aqueous NaCl and concentrated under reduced pressure, and the crude product was purified by automatic column chromatography (Biotage) (mobile phase: dichloromethane/0.5M ammonia in methanol 100/0 to 90/10) to give the title compound 23 as a white solid (24mg, 40%). LC-MS (ESI) M/z 564.0 (M + H) + 。 1 H NMR(400MHz,DMSO-d 6 )δ8.66(t,J＝5.9Hz,1H),7.86(s,1H),7.61(dd,J＝7.9,1.5Hz,1H),7.50–7.42(m,2H),7.41–7.34(m,2H),7.19(ddd,J＝8.5,7.0,1.6Hz,2H),7.02–6.93(m,2H),6.83(d,J＝8.2Hz,1H),6.63(d,J＝8.4Hz,1H),6.57–6.48(m,1H),6.20(d,J＝7.8Hz,1H),5.31(d,J＝7.4Hz,1H),4.96(q,J＝9.0Hz,2H),4.49(d,J＝5.9Hz,2H),3.61–3.52(m,1H),2.25–2.11(m,7H),1.86–1.74(m,4H),1.69–1.59(m,2H),1.57–1.47(m,2H).
EXAMPLE 24 synthetic route to Compound 24
Step 1: synthesis of Compound 24-1
To a solution of 5- (trifluoromethyl) -1H-pyrazole-3-carboxylic acid (72.74mg, 0.404mmol) in DMF (3 mL) were added DIPEA (94.91mg, 0.734mmol) and HATU (209.44mg, 0.551mmol) and the reaction was stirred at room temperature for 0.5H. Then 2-methylpropan-2-yl ({ 2- [3- (aminomethyl) phenyl)]-1- (2, 2-trifluoroethyl) indol-4-yl } [4- (dimethylamino) cyclohexyl]Carbamate (200mg, 0.367mmol), and the reaction stirred at room temperature for 1 hour. The reaction solution was diluted with ethyl acetate and water. The organic phase was separated and the aqueous phase was extracted with ethyl acetate. The combined organic phases were washed with water, saturated NaCl, dried over anhydrous Na2SO4, filtered, evaporated to dryness and purified by column chromatography (mobile phase: dichloromethane/0.5M amine methanol 0/1 to 10/1) to give compound 24-1 (90mg, 34.7%). LC-MS (ESI) M/z 707.4 (M + H) + .
Step 2: synthesis of Compound 24
To a reaction flask, compound 24-1 (90mg, 0.13mmol), DCM (3 mL), and TFA (0.6mL, 7.835mmol) were added, and the mixture was stirred at room temperature for 1 hour. The reaction solution was spin-dried, and subjected to prep-HPLC to obtain compound 24 (41mg, 51.3%). LC-MS (ESI) M/z 607.3 (M + H) + 。1H NMR(400MHz,DMSO-d6)δ9.29(t,J＝6.0Hz,1H),7.52–7.44(m,2H),7.39(dd,J＝12.5,7.6Hz,2H),7.32(s,1H),7.01–6.94(m,2H),6.84(d,J＝8.3Hz,1H),6.20(d,J＝7.8Hz,1H),5.34(d,J＝7.7Hz,1H),5.02(q,J＝9.1Hz,2H),4.58(d,J＝6.0Hz,2H),3.54(s,1H),2.19(s,6H),2.11(s,1H),1.78(d,J＝10.0Hz,4H),1.65(dd,J＝9.4,4.2Hz,2H),1.50(s,2H).
EXAMPLE 25 synthetic route to Compound 25
Step 1: synthesis of Compound 25-1
To a solution of 7-amino-2, 3-dihydro-1-indanone (0.32g, 2.174mmol) in DMF (20 mL) was added 2- [3- (bromomethyl) phenyl]-4, 5-tetramethyl-1, 3, 2-dioxaborane (4.52g, 15.219mmol), and the reaction was stirred at 80 ℃ for 2 hours. The reaction solution was diluted with ethyl acetate and water. The organic phase was separated and the aqueous phase was extracted with ethyl acetate. The organic phases were combined, washed with water, saturated NaCl, anhydrous Na 2 SO 4 Drying, filtering, evaporating to dryness, and purifying by column chromatography (mobile phase: petroleum ether/ethyl acetate 10/1 to 5/1) to obtain compound 25-1 (700mg, 88.6%). LC-MS (ESI) M/z 364.6 (M + H) +. (ii) a 1H NMR (400mhz, dmso-d 6) δ 7.75 (d, J =6.6hz, 1h), 7.67 (d, J =1.7hz, 1h), 7.55 (dt, J =7.2,1.3hz, 1h), 7.45 (dt, J =7.8,1.6hz, 1h), 7.38-7.26 (m, 2H), 6.60 (d, J =7.2hz, 1h), 6.41 (d, J =8.2hz, 1h), 4.48 (d, J =5.2hz, 2h), 2.99-2.91 (m, 2H), 2.64-2.54 (m, 2H), 1.28 (s, 12H).
Step 2: synthesis of Compound 25
To the reaction flask were added compound 25-1 (200mg, 0.43mmol), potassium carbonate (120mg, 0.86mmol), intermediate B (235mg, 0.645mmol), tetrakis (triphenylphosphine) palladium (50mg, 0.043mmol) and DMF (5 mL), water (1 mL). N for reaction solution 2 Replacement, stirring at 100 ℃Stirring for 1 hour. The reaction solution was diluted with ethyl acetate and water. The organic phase was separated and the aqueous phase was extracted with ethyl acetate. The organic phases were combined, washed with water, saturated NaCl, anhydrous Na 2 SO 4 Drying, filtration, evaporation to dryness and purification on column (mobile phase: dichloromethane/0.5M amine methanol 1/0 to 10/1) gave compound 25 (15mg, 88.6%). LC-MS (ESI) M/z 575.3 (M + H) + .1H NMR(400MHz,DMSO-d6)δ7.81(t,J＝6.3Hz,1H),7.50–7.43(m,2H),7.43–7.37(m,2H),7.29(t,J＝7.8Hz,1H),7.03–6.96(m,2H),6.90(d,J＝8.2Hz,1H),6.60(d,J＝7.3Hz,1H),6.45(d,J＝8.3Hz,1H),6.21(d,J＝7.7Hz,1H),5.24(d,J＝6.4Hz,1H),5.00(d,J＝9.1Hz,2H),4.58(d,J＝6.3Hz,2H),3.70(s,1H),2.94(t,J＝5.9Hz,2H),2.67–2.54(m,7H),2.04–1.93(m,2H),1.80(dd,J＝24.3,12.7Hz,4H),1.65(d,J＝12.7Hz,2H),1.23(s,2H).
EXAMPLE 26 synthetic route to Compound 26-p1 and Compound 26-p2
Step 1: synthesis of Compound 26-1
Add intermediate 1-1 (200mg, 0.479mmol), paraformaldehyde (72mg, 2.40mmol), methanol (5 mL), acetic acid (1 mL), naBH3CN (60mg, 0.958mmol) to the reaction flask. The mixture was stirred at 50 ℃ overnight. After the reaction, ethyl acetate and water were added to dilute. The organic phase was separated and the aqueous phase was extracted with ethyl acetate. The combined organic phases were washed with saturated aqueous NaCl and concentrated under reduced pressure to give crude compound 26-1 (186mg, 90%) which was used directly in the next reaction. LC-MS (ESI): M/z 432.1 (M + H) + 。
Synthesis of Compound 26-p1 and Compound 26-p2
Add Compound 26-1 (120mg, 0.257mmol), intermediate B (167mg, 0.386mmol), DMF (5 mL), water (1 mL), potassium carbonate (71mg, 0.515mmol) to the reaction flask, replace 3 to 5 times with nitrogen, add Pd (PPh) 3 ) 4 (30mg, 0.025mmol), and the nitrogen gas was further replaced 3 to 5 times. The mixture was stirred at 100 ℃ for 2 hours under nitrogen and then cooled to room temperature. Adding saturated EDTA aqueous solution and stirring at room temperature for 1 hourAnd adding ethyl acetate and water for dilution. The organic phase was separated and the aqueous phase was extracted with ethyl acetate. The organic phases were combined, washed with saturated aqueous NaCl solution, and concentrated under reduced pressure to afford crude product purification (Boston pHlex ODS, 21.2X 250mm,10um, water (0.05% trifluoroacetic acid)/acetonitrile, flow rate 30mL/min, column temperature 25 ℃, detection wavelength 254 nm) to afford the title compound 26-p1 as a white solid (13mg, 8%) and the title compound 26-p2 as a white solid (24mg, 14%). Compound 26-p1: LC-MS (ESI) M/z 643.0 (M + H) + 。 1 H NMR(400MHz,DMSO-d 6 ) δ 7.46 (t, J =7.6hz, 1h), 7.43-7.35 (m, 4H), 7.33 (d, J =7.5hz, 1h), 7.04-6.93 (m, 2H), 6.83 (d, J =9.6hz, 2h), 6.21 (d, J =7.8hz, 1h), 5.42 (d, J =8.1hz, 1h), 4.96 (q, J =9.0hz, 2h), 4.45 (s, 2H), 3.88 (s, 3H), 3.28 (s, 1H), 3.15 (s, 3H), 2.76 (s, 3H), 2.45-2.21 (m, 7H), 2.11 (d, J =12.3hz, 2h), 1.92 (d, J =12.0hz, 2h), 1.43 (q = 12.26H), 1.26H, 26H: LC-MS (ESI) M/z 643.0 (M + H) + 。 1 H NMR(400MHz,DMSO-d 6 )δ7.50–7.43(m,2H),7.42–7.36(m,3H),7.34(d,J＝7.5Hz,1H),7.05–6.96(m,3H),6.89(d,J＝8.3Hz,1H),6.22(d,J＝7.7Hz,1H),5.25(d,J＝6.5Hz,1H),4.98(q,J＝9.0Hz,2H),4.46(s,2H),3.89(s,3H),3.76–3.66(m,1H),3.15(s,3H),3.06–2.88(m,1H),2.77(s,3H),2.59(s,6H),2.04–1.95(m,2H),1.91–1.72(m,4H),1.69–1.59(m,2H).
EXAMPLE 27 synthetic route to Compound 27-p1 and Compound 27-p2
Step 1: synthesis of Compound 27-1
Add 4-nitroindole (3.00g, 18.50mmol), THF (50 mL) to the reaction flask. The mixture was cooled to 0 ℃ under nitrogen and NaH (1.48g, 37.00mmol) was added. After the addition, the temperature is raised to room temperature and the mixture is stirred for 0.5 hour, CF is added 3 CH 2 OTf (8.59g, 37.00mmol). After the reaction mixture was stirred at room temperature for 2 hours, the reaction mixture was cooled to 0 ℃. Slowly adding water to quench the reaction, and then adding ethyl acetate and water to dilute. The organic phase was separated and the aqueous phase was extracted with ethyl acetate. MergingOrganic phase, washing with saturated aqueous NaCl solution, anhydrous Na 2 SO 4 Drying, filtration and concentration under reduced pressure gave the title compound 27-1 (4.06g, 90%) as a yellow solid. 1 H NMR(400MHz,Chloroform-d)δ8.19(d,J＝8.2Hz,1H),8.13(dd,J＝8.0,0.8Hz,1H),7.82(d,J＝3.2Hz,1H),7.45(t,J＝8.1Hz,1H),7.17(dd,J＝3.2,0.9Hz,1H),5.41(q,J＝9.3Hz,2H).
Step 2: synthesis of Compound 27-2
To a reaction flask were added compound 27-1 (600mg, 2.46mmol), acetonitrile (10 mL), 4-imidazolecarboxaldehyde (472mg, 4.91mmol), TBHP (682. Mu.L, 4.91 mmol), and iodine (125mg, 0.491mmol). The mixture was stirred in a microwave reactor at 100 ℃ for 15 hours and then cooled to room temperature. Ethyl acetate and water were added for dilution. The organic phase was separated and the aqueous phase was extracted with ethyl acetate. The combined organic phases were washed with saturated aqueous NaCl solution and concentrated under reduced pressure, and the crude product was purified by automatic column chromatography (Biotage) (mobile phase: petroleum ether/ethyl acetate 100/0 to 50/50) to give the title compound 27-2 as a grey solid (40mg, 5%). LC-MS (ESI) M/z 338.7 (M + H) + .1H NMR(400MHz,DMSO-d 6 )δ9.88(s,1H),8.57(d,J＝1.2Hz,1H),8.38(d,J＝8.3Hz,1H),8.34(d,J＝1.2Hz,1H),8.26(dd,J＝8.1,0.7Hz,1H),7.63(t,J＝8.1Hz,1H),7.48(d,J＝0.8Hz,1H),5.31(q,J＝9.0Hz,2H).
And 3, step 3: synthesis of Compound 27-3
To the reaction flask were added compound 27-2 (100mg, 0.295mmol), anhydrous methanol (6 mL), N- (4-amino-3-methoxyphenyl) methanesulfonamide (119mg, 0.591mmol), acetic acid (220. Mu.L, 3.84 mmol), sodium cyanoborohydride (37mg, 0.591mmol). The mixture was stirred at 50 ℃ for 4 hours under nitrogen and then cooled to room temperature. After the reaction, the reaction solution is directly decompressed and concentrated to remove most of methanol, and ethyl acetate and water are added for dilution. The organic phase was separated and the aqueous phase was extracted with ethyl acetate. The combined organic phases were washed with saturated aqueous NaCl solution and concentrated under reduced pressure, and the crude product was purified by automatic column chromatography (Biotage) (mobile phase: petroleum ether/ethyl acetate 100/0 to 20/80) to give the title compound 27-3 (138mg, 64%) as a yellow solid. LC-MS (ESI) M/z 524.1 (M + H) + .
And 4, step 4: synthesis of Compound 27-4
To a reaction flask, compound 27-3 (120mg, 0.229 mmol), ethanol (9 mL), saturated aqueous ammonium chloride (3 mL), and iron powder (77mg, 1.38mmol) were added. After the mixture was stirred at 70 ℃ for 1 hour, the reaction solution was cooled to room temperature. The reaction solution was diluted with ethyl acetate and water. The organic phase was separated and the aqueous phase was extracted with ethyl acetate. The combined organic phases were washed with saturated aqueous NaCl, concentrated under reduced pressure and purified by an automatic column chromatography (Biotage) (mobile phase: petroleum ether/ethyl acetate 100/0 to 0/100) to give the title compound 27-4 (78mg, 69%) as a pale yellow solid. LC-MS (ESI): M/z 494.0 (M + H) + .
And 5: compound 27-p1 and compound 27-p2
To a reaction flask were added compound 27-4 (50mg, 0.101mmol), absolute ethanol (3 mL), tetrahydrofuran (3 mL), 4-dimethylaminocyclohexanone (71mg, 0.506 mmol), ethyl titanate (116mg, 0.506 mmol). The mixture was stirred at 50 ℃ for 5 hours under nitrogen and then cooled to 0 ℃. Sodium cyanoborohydride (32mg, 0.506mmol) was added and the mixture was stirred at 0 ℃ for 5 minutes, then warmed to 50 ℃ and stirred for 1 hour. After the reaction is finished, the reaction liquid is directly decompressed and concentrated to remove most of ethanol, and ethyl acetate and water are added for dilution. The organic phase was separated and the aqueous phase was extracted with ethyl acetate. The organic phases were combined, washed with saturated aqueous NaCl solution, and concentrated under reduced pressure to purify the crude product (Boston pHlex ODS, 21.2X 250mm,10um, water (0.05% trifluoroacetic acid)/acetonitrile, flow rate 30mL/min, column temperature 25 ℃ C., detection wavelength 254 nm) to obtain the title compound 27-p1 (13mg, 15%) as a white solid and the title compound 27-p2 (16mg, 18%) as a white solid.
Compound 27-p1 spectral data: LC-MS (ESI) M/z 619.0 (M + H) + 。 1 H NMR(400MHz,DMSO-d 6 )δ7.86(d,J＝1.3Hz,1H),7.33(d,J＝1.3Hz,1H),7.30(dd,J＝8.4,2.0Hz,1H),7.20(d,J＝2.1Hz,1H),7.05(t,J＝8.0Hz,1H),6.94(s,1H),6.81(d,J＝8.3Hz,1H),6.75(d,J＝8.4Hz,1H),6.27–6.20(m,2H),5.46(d,J＝8.1Hz,1H),4.77(q,J＝9.1Hz,2H),4.35(d,J＝5.7Hz,2H),3.89(s,3H),3.30–3.28(m,1H),3.07(s,3H),2.25–2.12(m,7H),2.10–2.02(m,2H),1.88–1.78(m,2H),1.35–1.27(m,4H).
Compound 27-p1 spectral data: LC-MS (ESI) M/z 619.0 (M + H) + 。 1 H NMR(400MHz,DMSO-d 6 )δ7.86(d,J＝1.3Hz,1H),7.34(d,J＝1.3Hz,1H),7.30(dd,J＝8.4,2.0Hz,1H),7.20(d,J＝2.0Hz,1H),7.08–7.02(m,2H),6.81(d,J＝8.3Hz,1H),6.75(d,J＝8.5Hz,1H),6.29–6.19(m,2H),5.40(d,J＝7.7Hz,1H),4.77(q,J＝9.2Hz,2H),4.36(d,J＝5.8Hz,2H),3.89(s,3H),3.60–3.48(m,1H),3.07(s,3H),2.16(s,6H),2.07–2.01(m,1H),1.85–1.69(m,4H),1.68–1.58(m,2H),1.52–1.44(m,2H).
EXAMPLE 28 synthetic route to Compound 28
Step 1: synthesis of Compound 28-1
To the reaction flask was added 7-amino-2, 3-dihydro-1-indanone (600mg, 4.08mmol), 1,4-dioxane (3 mL), tert-butylsulfinamide (1.48g, 12.23mmol), ethyl titanate (4.65g, 20.38mmol). The mixture was stirred at 115 ℃ overnight. After cooling to room temperature, ethyl acetate and water were added for dilution. The organic phase was separated and the aqueous phase was extracted with ethyl acetate. The combined organic phases were washed with saturated aqueous NaCl and concentrated under reduced pressure, and the crude product was purified by autosampler (Biotage) (mobile phase: petroleum ether/ethyl acetate 100/0 to 40/60) to give target compound 28-1 as a brown solid (107mg, 10%). LC-MS (ESI) M/z 251.1 (M + H) + .
Step 2: synthesis of Compound 28-2
Add 28-1 (100mg, 0.399mmol), THF (10 mL) to the flask. After the mixture was cooled to-78 deg.C, DIBAL-H (2.4mL, 2.4mmol) was added dropwise. After dropping, the mixture was slowly warmed to room temperature and stirred. After the reaction, water quenching is added to quench the reaction, and ethyl acetate is used for dilution. The organic phase was separated and the aqueous phase was extracted with ethyl acetate. The combined organic phases were washed with saturated aqueous NaCl solution and concentrated under reduced pressure, and the crude product was purified by automatic column chromatography (Biotage) (mobile phase: petroleum ether/ethyl acetate 100/0 to 40/60) to give the title compound 28-2 as a brown oil (65mg, 64%). LC-MS (ESI) M/z 253.1 (M + H) + .
And step 3: synthesis of Compound 28-3
Adding (3-bromomethyl) into a reaction bottlePhenylphenyl) boronic acid pinacol ester (70mg, 0.235mmol), DMF (5 mL), compound 28-2 (65mg, 0.259mmol), potassium carbonate (98mg, 0.707mmol). The mixture was stirred at 70 ℃ for 2 hours. After the reaction, ethyl acetate and water were added to dilute the reaction mixture. The organic phase was separated and the aqueous phase was extracted with ethyl acetate. The organic phases were combined, washed with saturated aqueous NaCl solution, concentrated under reduced pressure to give crude compound 28-3 (111 mg,>99%) was used in the next reaction without purification. LC-MS (ESI) M/z 469.3 (M + H) + .
And 4, step 4: synthesis of Compound 28-4
Add compound 28-3 (60mg, 0.128mmol), intermediate B (109mg, 0.232mmol), DMF (5 mL), water (1 mL), potassium carbonate (36mg, 0.257mmol) to the reaction flask, replace 3 to 5 times with nitrogen, add Pd (PPh) 3 ) 4 (15mg, 0.012mmol), and nitrogen was further substituted 3 to 5 times. The mixture was stirred at 100 ℃ for 2 hours under nitrogen and then cooled to room temperature. Add ethyl acetate and water to dilute. The organic phase was separated and the aqueous phase was extracted with ethyl acetate. The combined organic phases were washed with saturated aqueous NaCl and concentrated under reduced pressure, and the crude product was purified by automatic column chromatography (Biotage) (mobile phase: dichloromethane/0.5M ammonia in methanol 100/0 to 90/10) to give target compound 28-4 (51mg, 58%) as a white solid. LC-MS (ESI) M/z 680.3 (M + H) + .
And 5: synthesis of Compound 28
To a reaction flask, compound 28-4 (50mg, 0.073mmol), 1,4-dioxane (2.5 mL), methanol (2.5 mL), hydrogen chloride-1, 4-dioxane (0.5mL, 4M) were added. After the mixture was stirred at room temperature for 3 hours, a saturated aqueous sodium hydrogen carbonate solution was added to neutralize the reaction, and the reaction mixture was diluted with ethyl acetate. The organic phase was separated and the aqueous phase was extracted with ethyl acetate. The combined organic phases were washed with saturated aqueous NaCl solution and concentrated under reduced pressure to afford crude purified (Boston pHlex ODS, 21.2X 250mm,10um, water (0.05% trifluoroacetic acid)/acetonitrile, flow rate 30mL/min, column temperature 25 ℃ C., detection wavelength 254 nm) to afford target compound 28 (12mg, 28%) as a white solid. LC-MS (ESI) M/z 576.3 (M + H) + .1H NMR(400MHz,DMSO-d 6 )δ8.28(s,2H),7.57–7.48(m,2H),7.45(t,J＝7.5Hz,1H),7.37(dt,J＝7.4,1.6Hz,1H),7.04–6.97(m,3H),6.91(d,J＝8.2Hz,1H),6.76(t,J＝6.0Hz,1H),6.51(d,J＝7.3Hz,1H),6.28(d,J＝8.1Hz,1H),6.22(d,J＝7.7Hz,1H),5.19(d,J＝6.5Hz,1H),5.02(h,J＝8.2Hz,2H),4.79(d,J＝7.4Hz,1H),4.53–4.37(m,2H),3.80–3.70(m,1H),3.21–3.05(m,2H),2.68(s,7H),2.42–2.29(m,1H),2.12(dd,J＝14.1,7.8Hz,1H),2.05–1.98(m,2H),1.91–1.79(m,4H),1.69–1.59(m,2H).
EXAMPLE 29 synthetic route to Compound 29-p1 and Compound 29-p2
Step 1: synthesis of Compound 29-1
To the reaction flask were added (3-bromomethylphenyl) boronic acid pinacol ester (1 g, 3.37mmol), DMF (20 mL), 2-aminobenzonitrile (796 mg, 6.73mmol), potassium carbonate (1.4g, 10.10mmol). The mixture was stirred at 70 ℃ for 3 hours. After the reaction, ethyl acetate and water were added to dilute the reaction mixture. The organic phase was separated and the aqueous phase was extracted with ethyl acetate. The combined organic phases were washed with saturated aqueous NaCl solution and concentrated under reduced pressure, and the crude product was purified by automatic column chromatography (Biotage) (mobile phase: petroleum ether/ethyl acetate 100/0 to 85/15) to give the title compound 29-1 as a white solid (125mg, 11%). LC-MS (ESI) M/z 335.1 (M + H) + .
Step 2: synthesis of Compounds 29-p1 and 29-p2
To a reaction flask, intermediate B (110mg, 0.236 mmol), compound 29-1 (119mg, 0.354mmol), DMF (1 mL), water (1 mL), potassium carbonate (65mg, 0.472mmol) were added, and replacement with nitrogen was carried out 3 to 5 times, followed by addition of Pd (PPh) 3 ) 4 (27mg, 0.023mmol), nitrogen was replaced 3 to 5 times. The mixture was stirred at 100 ℃ for 2 hours under nitrogen and then cooled to room temperature. Add ethyl acetate and water to dilute. The organic phase was separated and the aqueous phase was extracted with ethyl acetate. The organic phases were combined, washed with saturated aqueous NaCl, concentrated under reduced pressure and the crude product purified (Boston pHlex ODS, 21.2X 250mm,10um, water (0.05% trifluoroacetic acid)/acetonitrile, flow rate 30mL/min, column temperature 25 ℃, detection wavelength 254 nm) to give the title compound 29-p1 (24mg, 19%) and the title compound 29-p2 (37mg, 29%) as white solids.
29-p1：LC-MS(ESI):m/z 546.3(M+H) + 。 1 H NMR(400MHz,DMSO-d 6 )δ7.49–7.43(m,3H),7.41(dt,J＝7.8,1.6Hz,1H),7.35(dt,J＝7.3,1.7Hz,1H),7.32–7.26(m,1H),7.02–6.90(m,2H),6.84(d,J＝7.8Hz,2H),6.66–6.58(m,2H),6.20(d,J＝7.8Hz,1H),5.40(d,J＝8.0Hz,1H),4.96(q,J＝9.0Hz,2H),4.51(d,J＝6.1Hz,2H),3.31–3.28(m,1H),2.42–2.25(m,7H),2.14–2.04(m,2H),1.94–1.83(m,2H),1.44–1.25(m,4H).
29-p2：LC-MS(ESI):m/z 546.3(M+H) + 。 1 H NMR(400MHz,DMSO-d 6 )δ7.50–7.43(m,3H),7.41(dt,J＝7.7,1.6Hz,1H),7.36(dt,J＝7.3,1.7Hz,1H),7.33–7.26(m,1H),7.01–6.90(m,3H),6.85(d,J＝8.2Hz,1H),6.68–6.58(m,2H),6.20(d,J＝7.7Hz,1H),5.30(d,J＝7.3Hz,1H),4.97(q,J＝9.0Hz,2H),4.51(d,J＝6.1Hz,2H),3.60–3.53(m,1H),2.25(s,7H),1.89–1.73(m,4H),1.69–1.60(m,2H),1.59–1.49(m,2H).
EXAMPLE 30 synthetic route to Compound 30-p1 and Compound 30-p2
Step 1: synthesis of Compound 30-1
To a reaction flask, compound 4-4 (90mg, 0.157mmol), 4-dimethylaminocyclohexanone (67mg, 0.472mmol), ethanol (5 mL), ethyl titanate (180mg, 0.787 mmol) were added, and the mixture was stirred overnight at 50 ℃ under nitrogen and then cooled to room temperature. Sodium cyanoborohydride (99mg, 1.570 mmol) was added. The mixture was stirred at 80 ℃ for 1 hour under nitrogen and then cooled to room temperature. Add ethyl acetate and water to dilute. The organic phase was separated and the aqueous phase was extracted with ethyl acetate. The combined organic phases were washed with saturated aqueous NaCl solution and concentrated under reduced pressure, and the crude product was purified by an automatic column chromatography (Biotage) (mobile phase: dichloromethane/0.5M ammonia in methanol 100/0 to 90/10) to give the title compound 30-1 as a white solid (50mg, 46%). LC-MS (ESI) M/z 697.3 (M + H) + .
Step 2: synthesis of Compound 30-p1 and Compound 30-p2
To a reaction flask were added compound 30-1 (50mg, 0.071mmol), dichloromethane (6 mL), and trifluoroacetic acid (3 mL), and the mixture was stirred at room temperature for 1 hour, then neutralized with saturated aqueous sodium bicarbonate solution, and diluted with dichloromethane. The organic phase was separated and the aqueous phase was extracted with dichloromethane. The organic phases were combined, washed with saturated aqueous NaCl solution, and concentrated under reduced pressure to obtain crude product purified (Boston pHlex ODS, 21.2X 250mm,10um, water (0.05% trifluoroacetic acid)/acetonitrile, flow rate 30mL/min, column temperature 25 ℃ C., detection wavelength 254 nm) to obtain target compound 30-p1 (11mg, 26%) as a white solid and target compound 30-p2 (15mg, 35%) as a white solid.
Compound 30-p1: LC-MS (ESI) M/z 597.3 (M + H) + 。 1 H NMR(400MHz,DMSO-d 6 )δ7.94–7.88(m,1H),7.83(d,J＝8.4Hz,1H),7.60(t,J＝7.8Hz,1H),7.49(d,J＝1.9Hz,1H),7.43(dt,J＝7.8,2.0Hz,2H),7.29(dd,J＝8.4,2.0Hz,1H),7.21(d,J＝2.0Hz,1H),7.11(dt,J＝7.2,3.7Hz,2H),7.07–7.02(m,1H),6.79(t,J＝6.4Hz,1H),6.64–6.55(m,2H),6.47(d,J＝1.9Hz,1H),5.65(d,J＝7.9Hz,1H),4.57(d,J＝6.4Hz,2H),3.88(s,3H),3.18–3.11(m,1H),3.06(s,3H),2.37–2.21(m,7H),2.08–2.01(m,2H),1.89–1.80(m,2H),1.36–1.29(m,2H),1.19–1.09(m,2H).
Compound 30-p2: LC-MS (ESI) M/z 597.3 (M + H) + 。 1 H NMR(400MHz,DMSO-d 6 )δ7.94–7.88(m,1H),7.83(d,J＝8.5Hz,1H),7.60(t,J＝7.7Hz,1H),7.50(d,J＝1.8Hz,1H),7.48–7.41(m,2H),7.28(dd,J＝8.4,2.0Hz,1H),7.21(d,J＝2.0Hz,1H),7.16–7.09(m,2H),7.05(dt,J＝5.5,3.1Hz,1H),6.80(t,J＝6.4Hz,1H),6.65(dd,J＝8.5,1.9Hz,1H),6.61(d,J＝8.5Hz,1H),6.47(d,J＝1.9Hz,1H),5.75(d,J＝7.4Hz,1H),4.56(d,J＝6.4Hz,2H),3.88(s,3H),3.06(s,3H),2.28(s,7H),1.77–1.62(m,4H),1.56–1.46(m,4H).
Example 31: synthetic routes to Compound 31-p1 and Compound 31-p2
Step 1: synthesis of Compound 31-1
Adding (3-Bromomethylphenyl) boronic acid pinacol ester (700mg, 2.357mmol), DMF (10 mL), 8-aminoisoquinoline (509.73mg, 3.535mmol), potassium carbonate (977.17mg, 7.071mmol). The mixture was stirred at 70 ℃ for 2 hours. After the reaction, ethyl acetate and water were added to dilute the reaction mixture. The organic phase was separated and the aqueous phase was extracted with ethyl acetate. The combined organic phases were washed with saturated aqueous NaCl solution and concentrated under reduced pressure to give crude compound 31-1 (866 mg,>99%) was used in the next reaction without purification. LC-MS (ESI) M/z 361.2 (M + H) + .
Step 2: synthesis of Compound 31-p1 and Compound 31-p2
Add 31-1 (200mg, 0.430mmol), intermediate B (464.55mg, 1.289mmol), DMF (5 mL), water (1 mL), potassium carbonate (118.81mg, 0.860mmol), nitrogen exchange 3-5 times, add Pd (PPh 3) 4 (49.69mg, 0.043mmol), and nitrogen exchange 3-5 times. The mixture was stirred at 100 ℃ for 2 hours under nitrogen and then cooled to room temperature. Add ethyl acetate and water to dilute. The organic phase was separated and the aqueous phase was extracted with ethyl acetate. The organic phases were combined, washed with saturated aqueous NaCl solution, and concentrated under reduced pressure to obtain crude product purified (Boston pHlex ODS, 21.2X 250mm,10um, water (0.05% trifluoroacetic acid)/acetonitrile, flow rate 30mL/min, column temperature 25 ℃ C., detection wavelength 254 nm) to obtain 31-p1 (10mg, 4%) as a white solid and 31-p2 (18mg, 7%) as a white solid.
Compound 31-p1: LC-MS (ESI) M/z 572.4 (M + H) + 。 1 H NMR(400MHz,DMSO-d 6 )δ9.64(s,1H),8.40(d,J＝5.6Hz,1H),7.66–7.59(m,2H),7.54(s,1H),7.51–7.43(m,2H),7.42–7.34(m,2H),7.04(d,J＝8.0Hz,1H),6.96(t,J＝7.9Hz,1H),6.87–6.80(m,2H),6.51(d,J＝7.8Hz,1H),6.19(d,J＝7.8Hz,1H),5.39(d,J＝8.1Hz,1H),4.95(q,J＝9.1Hz,2H),4.61(d,J＝5.9Hz,2H),2.33(s,7H),2.11–2.05(m,2H),1.92–1.85(m,2H),1.49–1.34(m,4H).
Compound 31-p2: LC-MS (ESI) M/z 572.3 (M + H) + 。 1 H NMR(400MHz,DMSO-d 6 )δ9.64(s,1H),8.40(d,J＝5.7Hz,1H),7.66–7.58(m,2H),7.54(s,1H),7.49–7.43(m,2H),7.41–7.35(m,2H),7.04(d,J＝8.1Hz,1H),6.99–6.92(m,2H),6.82(d,J＝8.3Hz,1H),6.52(d,J＝7.8Hz,1H),6.19(d,J＝7.7Hz,1H),5.29(d,J＝7.4Hz,1H),4.95(q,J＝9.0Hz,2H),4.61(d,J＝5.8Hz,2H),3.59–3.50(m,1H),2.28–2.15(m,7H),1.83–1.75(m,4H),1.67–1.60(m,2H),1.55–1.48(m,2H).
EXAMPLE 32 synthetic route to Compound 32-p1 and Compound 32-p2
Step 1: synthesis of Compound 32-1
Into the reaction flask was added 8-bromoisochromane (200mg, 0.939mmol), 1,4-dioxane (10 mL), tert-butyl carbamate (219.93mg, 1.877mmol), cs2CO3 (611.67mg, 1.877mmol), XPHOS (89.50mg, 0.188mmol). The mixture was purged 3 to 5 times with nitrogen, pd (OAc) 2 (21.07mg, 0.094mmol) was added, and the mixture was purged 3 to 5 times with nitrogen. The mixture was stirred at 100 ℃ for 2 hours and then cooled to room temperature. After the reaction, ethyl acetate and water were added to dilute the reaction mixture. The organic phase was separated and the aqueous phase was extracted with ethyl acetate. The combined organic phases were washed with saturated aqueous NaCl solution and concentrated under reduced pressure, and the crude product was purified by automatic column chromatography (Biotage) (mobile phase: petroleum ether/ethyl acetate 100/0 to 89/11) to give the title compound 32-1 as a brown solid (225mg, 77%). LC-MS (ESI) M/z 194.1 (M-tBu) + .
Step 2: synthesis of Compound 32-2
To the reaction flask was added (3-bromomethylphenyl) boronic acid pinacol ester (238.26mg, 0.802mmol), DMF (15 mL), compound 32-1 (100mg, 0.401mmol), cs 2 CO 3 (391.96mg, 1.203mmol). The mixture was stirred at 50 ℃ for 2 hours. After the reaction, ethyl acetate and water were added to dilute the reaction mixture. The organic phase was separated and the aqueous phase was extracted with ethyl acetate. The combined organic phases were washed with saturated aqueous NaCl and concentrated under reduced pressure, and the crude product was purified by autosampler (Biotage) (mobile phase: petroleum ether/ethyl acetate 100/0 to 82/18) to give the title compound 32-2 as a brown solid (146mg, 78%). LC-MS (ESI): M/z 466.2 (M + H) + .
And step 3: synthesis of Compound 32-3
Adding the intermediate B (110mg, 0.236 mmol) and the compound 32-2 (143.03mg, 0.307mmol) into a reaction bottle,DMF (5 mL), water (1 mL), potassium carbonate (65.34mg, 0.473mmol), displacement with nitrogen 3 to 5 times, addition of Pd (PPh) 3 ) 4 (27.32mg, 0.024mmol), nitrogen was replaced 3 to 5 times. The mixture was stirred at 100 ℃ for 2 hours under nitrogen and then cooled to room temperature. Add ethyl acetate and water to dilute. The organic phase was separated and the aqueous phase was extracted with ethyl acetate. The combined organic phases were washed with saturated aqueous NaCl and concentrated under reduced pressure, and the crude product was purified by automatic column chromatography (Biotage) (mobile phase: dichloromethane/0.5M ammonia in methanol 100/0 to 88/12) to give the title compound 32-3 (132mg, 82%) as a brown oil. LC-MS (ESI) M/z 677.4 (M + H) + .
And 4, step 4: synthesis of Compound 32-p1 and Compound 32-p2
Add compound 32-3 (130mg, 0.192mmol), dichloromethane (10 mL), trifluoroacetic acid (5 mL) to the reaction flask. After the mixture was stirred at room temperature for 1 hour, a saturated aqueous sodium bicarbonate solution was added to neutralize the reaction, and the reaction mixture was diluted with dichloromethane. The organic phase was separated and the aqueous phase was extracted with dichloromethane. The organic phases were combined, washed with saturated aqueous NaCl solution, and concentrated under reduced pressure to purify the crude product (Boston pHlex ODS, 21.2X 250mm,10um, water (0.05% trifluoroacetic acid)/acetonitrile, flow rate 30mL/min, column temperature 25 ℃, detection wavelength 254 nm) to obtain the target compound 32-p1 (28mg, 25%) as a white solid and the target compound 32-p2 (38mg, 34%) as a white solid.
Compound 32-p1: LC-MS (ESI) M/z 577.3 (M + H) + 。 1 H NMR(400MHz,DMSO-d 6 )δ7.46–7.37(m,3H),7.33(dt,J＝7.4,1.6Hz,1H),6.97(t,J＝8.0Hz,1H),6.88–6.80(m,3H),6.35(d,J＝7.5Hz,1H),6.27(d,J＝8.1Hz,1H),6.19(d,J＝7.8Hz,1H),5.59(t,J＝6.0Hz,1H),5.40(d,J＝8.1Hz,1H),4.95(q,J＝9.0Hz,2H),4.58(s,2H),4.40(d,J＝5.9Hz,2H),3.80(t,J＝5.6Hz,2H),3.30–3.28(m,1H),2.68(t,J＝5.6Hz,2H),2.30(s,7H),2.14–2.05(m,2H),1.93–1.84(m,2H),1.44–1.26(m,4H).
Compound 32-p2: LC-MS (ESI) M/z 577.3 (M + H) + 。 1 H NMR(400MHz,DMSO-d 6 )δ7.46–7.37(m,3H),7.33(dt,J＝7.3,1.7Hz,1H),7.01–6.92(m,2H),6.84(t,J＝7.7Hz,2H),6.35(d,J＝7.5Hz,1H),6.27(d,J＝8.0Hz,1H),6.20(d,J＝7.7Hz,1H),5.59(t,J＝6.0Hz,1H),5.29(d,J＝7.3Hz,1H),4.96(q,J＝9.0Hz,2H),4.59(s,2H),4.40(d,J＝5.9Hz,2H),3.80(t,J＝5.6Hz,2H),3.62–3.52(m,1H),2.68(t,J＝5.6Hz,2H),2.38–2.15(m,7H),1.89–1.74(m,4H),1.68–1.50(m,4H).
Example 33: synthetic routes to Compound 33-p1 and Compound 33-p2
Step 1: synthesis of Compound 33-1
Into the reaction flask were charged N-BOC-8-bromo-1, 2,3, 4-tetrahydroisoquinoline (1.6g, 5.125mmol), 1,4-dioxane (30 mL), t-butyl carbamate (1.20g, 10.250mmol), cs 2 CO 3 (3.34g, 10.250mmol), XPHOS (0.49g, 1.025mmol). The mixture was purged with nitrogen 3 to 5 times, and Pd (OAc) was added 2 (0.12g, 0.512mmol), and the mixture was further purged with nitrogen 3 to 5 times. The mixture was stirred at 100 ℃ for 2 hours and then cooled to room temperature. After the reaction, ethyl acetate and water were added to dilute the reaction mixture. The organic phase was separated and the aqueous phase was extracted with ethyl acetate. The combined organic phases were washed with saturated aqueous NaCl solution and concentrated under reduced pressure, and the crude product was purified by automatic column chromatography (Biotage) (mobile phase: petroleum ether/ethyl acetate 100/0 to 82/18) to give the title compound 33-1 as a brown solid (1.418g, 79%). LC-MS (ESI) M/z 349.3 (M + H) + .
Step 2: synthesis of Compound 33-2
To a reaction flask were added compound 33-1 (400mg, 1.148mmol), DMF (10 mL), 3-bromobenzyl bromide (573.83mg, 2.296 mmol), cs 2 CO 3 (1122.10mg, 3.444mmol). The mixture was stirred at 50 ℃ for 3 hours. After the reaction, ethyl acetate and water were added to dilute. The organic phase was separated and the aqueous phase was extracted with ethyl acetate. The combined organic phases were washed with saturated aqueous NaCl solution and concentrated under reduced pressure, and the crude product was purified by automatic column chromatography (Biotage) (mobile phase: petroleum ether/ethyl acetate 100/0 to 82/18) to give the title compound 33-2 (552mg, 93%) as a colorless oil. LC-MS (ESI): M/z 517.0 (M + H) + .
And step 3: synthesis of Compound 33-3
Adding into a reaction flaskN, N' -dimethylethylenediamine (83.04mg, 0.942mmol), DMF (15 mL), K 2 CO 3 (390.75mg, 2.827mmol), nitrogen substitution of the mixture 3 to 5 times, cuI (179.49mg, 0.942mmol), stirring the mixture at room temperature for 15 minutes, addition of 2-nitrocarbazole (200mg, 0.942mmol), compound 33-2 (536.46mg, 1.037mmol), and nitrogen substitution of the mixture 3 to 5 times. The mixture was stirred overnight at 140 ℃ under nitrogen and allowed to cool to room temperature. Add ethyl acetate and water to dilute. The organic phase was separated and the aqueous phase was extracted with ethyl acetate. The combined organic phases were washed with saturated aqueous NaCl solution and concentrated under reduced pressure, and the crude product was purified by automatic column chromatography (Biotage) (mobile phase: petroleum ether/ethyl acetate 100/0 to 80/20) to give the desired compound 33-3 as a yellow oil (132mg, 21.59%). LC-MS (ESI) M/z 649.3 (M + H) + .
And 4, step 4: synthesis of Compound 33-4
Add Compound 33-3 (130mg, 0.200mmol), etOH (6 mL), NH to the reaction flask 4 Cl (aq.) (2 mL), fe (67.14mg, 1.202mmol). The mixture was stirred at 70 ℃ for 1 hour. After the reaction, the reaction solution was concentrated under reduced pressure and diluted with ethyl acetate and water. The organic phase was separated and the aqueous phase was extracted with ethyl acetate. The combined organic phases were washed with saturated aqueous NaCl solution and concentrated under reduced pressure, and the crude product was purified by automatic column chromatography (Biotage) (mobile phase: petroleum ether/ethyl acetate 100/0 to 40/60) to give the title compound 33-4 (86mg, 0.139mmol, 69.36%) as a yellow oil. LC-MS (ESI) M/z 619.8 (M + H) + .
And 5: synthesis of Compound 33-5
To the reaction flask were added compound 33-4 (80mg, 0.129mmol), etOH (5 mL), 4-dimethylaminocyclohexanone (91.08mg, 0.645mmol), ti (OEt) 4 (147.46mg, 0.646 mmol). The mixture was stirred at 50 ℃ overnight. The next day, the mixture was cooled to room temperature and NaBH3CN (81.24mg, 1.293mmol) was added and the mixture was warmed to 80 ℃ and stirred for 1 hour. After the reaction, ethyl acetate and water were added to dilute the reaction mixture. The organic phase was separated and the aqueous phase was extracted with ethyl acetate. The organic phases were combined, washed with saturated aqueous NaCl solution, concentrated under reduced pressure and the crude product was purified by automatic column chromatography (Biotage) (mobile phase: dichloromethane/0.5M ammonia in methanol 100/0 to 88/12) to give the title compound 33-5 (71mg, 0.095mmol, 73.81%) as a yellow oil. L is a radical of an alcoholC-MS(ESI):m/z 745.4(M+H) + .
Step 6: synthesis of Compounds 33-p1 and 33-p2
To a reaction flask, compound 33-5 (70mg, 0.094mmol), dichloromethane (6 mL), trifluoroacetic acid (2 mL) were added. After the mixture was stirred at room temperature for 1 hour, a saturated aqueous sodium bicarbonate solution was added to neutralize the reaction, and the reaction mixture was diluted with dichloromethane. The organic phase was separated and the aqueous phase was extracted with dichloromethane. The organic phases were combined, washed with saturated aqueous NaCl solution, and concentrated under reduced pressure to purify the crude product (Boston pHlex ODS, 21.2X 250mm,10um, water (0.05% trifluoroacetic acid)/acetonitrile, flow rate 30mL/min, column temperature 25 ℃, detection wavelength 254 nm) to obtain the target compound 33-p1 (15mg, 29%) as a white solid and the target compound 33-p2 (28mg, 55%) as a white solid.
Compound 33-p1: LC-MS (ESI) M/z 544.3 (M + H) + 。 1 H NMR(400MHz,DMSO-d 6 )δ7.94–7.88(m,1H),7.83(d,J＝8.4Hz,1H),7.59(t,J＝7.7Hz,1H),7.50(t,J＝1.8Hz,1H),7.46(d,J＝7.7Hz,1H),7.40(dt,J＝8.1,1.3Hz,1H),7.14–7.09(m,2H),7.09–7.04(m,1H),6.91(t,J＝7.8Hz,1H),6.58(dd,J＝8.5,1.9Hz,1H),6.44(d,J＝1.9Hz,1H),6.36(t,J＝7.8Hz,2H),5.71(t,J＝6.0Hz,1H),5.61(d,J＝8.0Hz,1H),4.47(d,J＝5.8Hz,2H),3.78(s,2H),3.16–3.09(m,1H),2.99(t,J＝5.9Hz,2H),2.69(t,J＝5.7Hz,2H),2.22–2.16(m,7H),2.06–2.00(m,2H),1.85–1.77(m,2H),1.34–1.26(m,2H),1.18–1.09(m,2H).
Compound 33-p2: LC-MS (ESI) M/z 544.3 (M + H) + 。 1 H NMR(400MHz,DMSO-d 6 )δ7.94–7.87(m,1H),7.82(d,J＝8.5Hz,1H),7.59(t,J＝7.7Hz,1H),7.52–7.44(m,2H),7.40(dt,J＝8.0,1.3Hz,1H),7.15–7.09(m,2H),7.09–7.03(m,1H),6.91(t,J＝7.8Hz,1H),6.64(dd,J＝8.5,1.9Hz,1H),6.47(d,J＝1.9Hz,1H),6.36(t,J＝8.7Hz,2H),5.79–5.65(m,2H),4.47(d,J＝5.8Hz,2H),3.78(s,2H),3.39–3.38(m,1H),2.99(t,J＝5.8Hz,2H),2.69(t,J＝5.8Hz,2H),2.19(s,6H),2.15–2.09(m,1H),1.74–1.60(m,4H),1.56–1.46(m,4H).
Example 34: synthetic routes to Compound 34-p1 and Compound 34-p2
Step 1: synthesis of Compound 34-1
Add intermediate B (140mg, 0.301mmol), compound 33-2 (509.57mg, 0.903mmol), DMF (5 mL), H to the flask 2 O(1mL)，K 2 CO 3 (83.20mg, 0.602mmol), the mixture was purged with nitrogen 3 to 5 times, and Pd (PPh) was added 3 ) 4 (34.77mg, 0.030mmol), the mixture was replaced with nitrogen 3 to 5 times. The mixture was stirred at 100 ℃ for 2 hours under nitrogen and then cooled to room temperature. Add ethyl acetate and water to dilute. The organic phase was separated and the aqueous phase was extracted with ethyl acetate. The combined organic phases were washed with saturated aqueous NaCl solution and concentrated under reduced pressure, and the crude product was purified by automatic column chromatography (Biotage) (mobile phase: dichloromethane/0.5M ammonia in methanol 100/0 to 90/10) to give target compound 34-1 (210mg, 90%) as a brown solid. LC-MS (ESI) M/z 776.4 (M + H) + .
Step 2: synthesis of Compound 34-p1 and Compound 34-p2
To the reaction flask were added compound 34-1 (190mg, 0.2450 mmol), dichloromethane (10 mL), trifluoroacetic acid (5 mL). After the mixture was stirred at room temperature for 1 hour, a saturated aqueous sodium bicarbonate solution was added to neutralize the reaction, and the reaction solution was diluted with dichloromethane. The organic phase was separated and the aqueous phase was extracted with dichloromethane. The organic phases were combined, washed with saturated aqueous NaCl solution, and concentrated under reduced pressure to purify the crude product (Boston pHlex ODS, 21.2X 250mm,10um, water (0.05% trifluoroacetic acid)/acetonitrile, flow rate 30mL/min, column temperature 25 ℃ C., detection wavelength 254 nm) to obtain target compound 34-p1 (30mg, 21%) as a white solid and target compound 34-p2 (54mg, 38%) as a white solid.
Compound 34-p1: LC-MS (ESI) M/z 576.3 (M + H) + 。 1 H NMR(400MHz,DMSO-d 6 )δ7.45–7.37(m,3H),7.33(dd,J＝7.1,1.9Hz,1H),6.97(t,J＝8.0Hz,1H),6.87–6.78(m,3H),6.32(d,J＝7.5Hz,1H),6.25(d,J＝8.1Hz,1H),6.19(d,J＝7.8Hz,1H),5.65(t,J＝6.0Hz,1H),5.37(d,J＝8.1Hz,1H),4.95(q,J＝9.0Hz,2H),4.41(d,J＝5.9Hz,2H),3.77(s,2H),3.29–3.24(m,2H),2.98(t,J＝5.8Hz,2H),2.66(t,J＝5.9Hz,2H),2.24–2.18(m,7H),2.11–2.05(m,2H),1.88–1.81(m,2H),1.40–1.25(m,4H).
Compound 34-p2: LC-MS (ESI) M/z 576.3 (M + H) + 。 1 H NMR(400MHz,DMSO-d 6 )δ7.46–7.37(m,3H),7.33(dt,J＝7.4,1.7Hz,1H),7.00–6.92(m,2H),6.87–6.78(m,2H),6.32(d,J＝7.5Hz,1H),6.25(d,J＝8.0Hz,1H),6.20(d,J＝7.8Hz,1H),5.64(t,J＝6.0Hz,1H),5.30(d,J＝7.5Hz,1H),4.95(q,J＝9.0Hz,2H),4.41(d,J＝5.9Hz,2H),3.76(s,2H),3.57–3.51(m,2H),2.96(t,J＝5.7Hz,2H),2.65(t,J＝5.8Hz,2H),2.18(s,6H),2.11–2.04(m,1H),1.84–1.72(m,4H),1.68–1.60(m,2H),1.54–1.46(m,2H).
Example 35: synthetic route to compound 35
Step 1: synthesis of Compound 35-1
Into the reaction flask was added N-BOC-8-bromo-1, 2,3, 4-tetrahydroisoquinoline (3g, 9.609mmol), 1,4-dioxane hydrochloride (30 mL). The mixture was stirred at room temperature for 1 hour. After the reaction, the reaction mixture was directly filtered under reduced pressure, and the obtained solid was further dried under reduced pressure to obtain the desired compound 35-1 (2.2g, 8.851mmol, 92.12%) as a white solid. LC-MS (ESI) M/z 212.0 (M + H) + .
And 2, step: synthesis of Compound 35-2
To a reaction flask was added compound 35-1 (2.2g, 8.851mmol), DMF (40 mL), K 2 CO 3 (3.67g, 26.554mmol). After the mixture was stirred at 0 ℃ for 5 minutes, methyl iodide (1.077mL, 13.277mmol) was slowly added dropwise, and after the addition, the mixture was slowly warmed to room temperature and stirred for 2 hours. After the reaction, water and ethyl acetate were added to dilute the reaction mixture. The organic phase was separated and the aqueous phase was extracted with ethyl acetate. The organic phases were combined, washed with a saturated aqueous NaCl solution, and concentrated under reduced pressure to give 35-2 (750mg, 3.317mmol, 37.47%) of the objective compound as a colorless liquid. LC-MS (ESI): M/z 226.0 (M + H) + .
And step 3: synthesis of Compound 35-3
Add Compound 35-2 (100mg, 0.184mmol), 2-methylpropan-2-yl ({ 2- [3 ] to the reaction flask- (aminomethyl) phenyl]-1- (2, 2-trifluoroethyl) indol-4-yl } [4- (dimethylamino) cyclohexyl]Amino) formate (41.52mg, 0.184mmol), toluene (5 mL), john Phos (21.96mg, 0.074mmol), tBuONa (26.47mg, 0.275mmol), nitrogen substitution of the mixture 3 to 5 times, addition of Pd 2 (dba) 3 (33.70mg, 0.037 mmol), and the mixture was further purged with nitrogen 3 to 5 times. The mixture was stirred overnight at 100 ℃ under nitrogen and allowed to cool to room temperature. The organic phase was separated and the aqueous phase was extracted with ethyl acetate. The organic phases were combined, washed with saturated aqueous NaCl solution, concentrated under reduced pressure and the crude product purified by automatic column chromatography (Biotage) (mobile phase: dichloromethane/0.5M ammonia in methanol 100/0 to 88/12) to give the title compound 35-3 as a brown oil (110mg, 25% (total yield)). LC-MS (ESI): M/z 690.5 (M + H) + .
And 4, step 4: synthesis of Compound 35
To the reaction flask were added compound 35-3 (100mg, 0.145mmol), dichloromethane (6 mL), trifluoroacetic acid (3 mL). After the mixture was stirred at room temperature for 1 hour, a saturated aqueous sodium bicarbonate solution was added to neutralize the reaction, and the reaction solution was diluted with dichloromethane. The organic phase was separated and the aqueous phase was extracted with dichloromethane. The organic phases were combined, washed with saturated aqueous NaCl, concentrated under reduced pressure and the crude product was purified (Boston pHlex ODS,21.2 x 250mm,10um, water (0.05% trifluoroacetic acid)/acetonitrile, flow rate 30mL/min, column temperature 25 ℃ C., detection wavelength 254 nm) to give target compound 35 (39mg, 46%) as a white solid. LC-MS (ESI) M/z 590.4 (M + H) + .
Compound 35 1 H NMR(400MHz,DMSO-d 6 )δ7.45–7.38(m,3H),7.33(dt,J＝7.1,1.8Hz,1H),6.97(t,J＝8.0Hz,1H),6.94(s,1H),6.85(d,J＝8.2Hz,1H),6.80(t,J＝7.8Hz,1H),6.33(d,J＝7.5Hz,1H),6.25(d,J＝8.1Hz,1H),6.20(d,J＝7.8Hz,1H),5.61(t,J＝6.0Hz,1H),5.29(d,J＝7.3Hz,1H),4.95(q,J＝9.1Hz,2H),4.40(d,J＝5.9Hz,2H),3.60–3.54(m,1H),3.35(s,2H),2.73(t,J＝5.8Hz,2H),2.55(t,J＝5.7Hz,2H),2.40(s,3H),2.26(s,7H),1.86–1.76(m,4H),1.67–1.59(m,2H),1.58–1.50(m,2H).
Example 36: synthetic route to compound 36
Step 1: synthesis of Compound 36-1
2-Methylpropan-2-yl 4-chloro-5, 6,7, 8-tetrahydropyrido [4,3-d ] was added to the reaction flask]Pyrimidine-6-carboxylic acid ester (300mg, 1.112mmol), DMF (6 mL), 3-aminomethylphenylboronic acid pinacol ester hydrochloride (359.80mg, 1.335mmol), cs 2 CO 3 (1087.15mg, 3.337mmol). The mixture was stirred at 100 ℃ for 3 hours with a microwave. After the reaction, ethyl acetate and water were added to dilute the reaction mixture. The organic phase was separated and the aqueous phase was extracted with ethyl acetate. The combined organic phases were washed with saturated aqueous NaCl solution and concentrated under reduced pressure, and the crude product was purified by automatic column chromatography (Biotage) (mobile phase: petroleum ether/ethyl acetate 100/0 to 60/40) to give target compound 36-1 (50mg, 0.107mmol, 9.64%) as a colorless oil. LC-MS (ESI) M/z 467.6 (M + H) + .
And 2, step: synthesis of Compound 36-2
Add Compound 36-1 (120.33mg, 0.258mmol), intermediate B (100mg, 0.215mmol), DMF (5 mL), water (1 mL), K to the reaction flask 2 CO 3 (59.43mg, 0.430mmol), nitrogen substitution 3 to 5 times, and addition of Pd (PPh) 3 ) 4 (24.85mg, 0.022mmol), and the nitrogen was further replaced 3 to 5 times. The mixture was stirred at 100 ℃ for 2 hours under nitrogen and then cooled to room temperature. Ethyl acetate and water were added for dilution. The organic phase was separated and the aqueous phase was extracted with ethyl acetate. The combined organic phases were washed with saturated aqueous NaCl solution and concentrated under reduced pressure, and the crude product was purified by column chromatography (Biotage) automatically (mobile phase: dichloromethane/0.5M ammonia in methanol 100/0 to 88/12) to give the title compound 36-2 as a brown oil (80mg, 0.118mmol, 54.92%). LC-MS (ESI) M/z 678.9 (M + H) + .
And 3, step 3: synthesis of Compound 36
Add compound 36-2 (100mg, 0.145mmol), dichloromethane (6 mL), trifluoroacetic acid (3 mL) to the reaction flask. After the mixture was stirred at room temperature for 1 hour, a saturated aqueous sodium bicarbonate solution was added to neutralize the reaction, and the reaction mixture was diluted with dichloromethane. The organic phase was separated and the aqueous phase was extracted with dichloromethane. The combined organic phases were washed with saturated aqueous NaCl solution and concentrated under reduced pressure to give a crude product which was purified (Boston pHlex ODS, 21.2X 250mm)10um, water (0.05% trifluoroacetic acid)/acetonitrile, flow rate 30mL/min, column temperature 25 ℃, detection wavelength 254 nm) to afford target compound 36 (22mg, 32%) as a white solid. LC-MS (ESI): M/z 578.4 (M + H) + .
Compound 36 1 H NMR(400MHz,DMSO-d 6 )δ8.20(s,1H),7.43(t,J＝7.5Hz,1H),7.38(s,1H),7.34(d,J＝7.5Hz,2H),7.26(t,J＝6.1Hz,1H),7.04–6.90(m,2H),6.83(d,J＝8.2Hz,1H),6.20(d,J＝7.8Hz,1H),5.32(d,J＝7.4Hz,1H),4.99(q,J＝9.0Hz,2H),4.67(d,J＝6.1Hz,2H),3.62–3.51(m,3H),2.94(t,J＝5.8Hz,2H),2.55–2.51(m,2H),2.20(s,6H),2.14–2.08(m,1H),1.86–1.73(m,4H),1.69–1.59(m,2H),1.55–1.45(m,2H).
Example 37: synthetic routes to Compounds 37-p1 and 37-p2
Step 1: synthesis of Compound 37-1
Add 8-bromo-5-fluoroisoquinoline (260mg, 1.150mmol), acOH (5 mL), naBH to the reaction flask 4 (152.29mg, 4.026mmol). The mixture was stirred at room temperature for 3 hours. After the reaction, the reaction mixture was concentrated under reduced pressure, and a saturated aqueous sodium bicarbonate solution and methylene chloride were added. The organic phase was separated and the aqueous phase was extracted with dichloromethane. The organic phases were combined, washed with a saturated aqueous NaCl solution, and concentrated under reduced pressure to give the desired compound 37-1 as a colorless oil (262mg, 1.139mmol, 99.00%). LC-MS (ESI) M/z 230.0 (M + H) + .
Step 2: synthesis of Compound 37-2
Add compound 37-1 (260mg, 1.130mmol), DCM (6 mL), DIEA (438.17mg, 3.390mmol), boc to the reaction flask 2 O (320.62mg, 1.469mmol). The mixture was stirred at 25 ℃ for 1.5 hours. Add ethyl acetate and water to dilute. The organic phase was separated and the aqueous phase was extracted with ethyl acetate. The combined organic phases were washed with saturated aqueous NaCl solution and concentrated under reduced pressure, and the crude product was purified by automatic column chromatography (Biotage) (mobile phase: petroleum ether/ethyl acetate 100/0 to 90/10) to give the desired compound 37-2 (306mg, 0.927mmol, 82.01%) as a colorless oil. LC-MS (ESI): M/z 276.0 (M-tBu) + .
And step 3: synthesis of Compound 37-3
To the reaction flask was added 2-methylpropan-2-yl ({ 2- [3- (aminomethyl) phenyl)]-1- (2, 2-trifluoroethyl) indol-4-yl } [4- (dimethylamino) cyclohexyl]Carbamate (175mg, 0.321mmol), compound 37-2 (137.79mg, 0.417mmol), cs 2 CO 3 (209.18mg,0.642mmol)，Pd 2 (dba) 3 (29.39mg, 0.032mmol), XPHOS (30.61mg, 0.064mmol), 1,4-dioxane (5 mL). After the mixture was purged with nitrogen 3 to 5 times, it was stirred at 100 ℃ overnight. After the reaction, ethyl acetate and water were added to dilute. The organic phase was separated and the aqueous phase was extracted with ethyl acetate. The combined organic phases were washed with saturated aqueous NaCl and concentrated under reduced pressure, and the crude product was purified by column chromatography (Biotage) (mobile phase: dichloromethane/0.5M ammonia in methanol 100/0 to 88/12) to give the title compound 37-3 as a pale yellow solid (192mg, 0.242mmol, 75.27%). LC-MS (ESI) M/z 794.4 (M + H) + .
And 4, step 4: synthesis of Compound 37-P1 and Compound 37-P2
Add compound 37-3 (210mg, 0.265mmol), DCM (12 mL), TFA (4 mL) to the reaction flask. After the mixture was stirred at room temperature for 1 hour, a saturated aqueous sodium bicarbonate solution was added to neutralize the reaction, and the reaction mixture was diluted with dichloromethane. The organic phase was separated and the aqueous phase was extracted with dichloromethane. The organic phases were combined, washed with saturated aqueous NaCl solution, and concentrated under reduced pressure to purify the crude product (Boston pHlex ODS, 21.2X 250mm,10um, water (0.05% trifluoroacetic acid)/acetonitrile, flow rate 30mL/min, column temperature 25 ℃ C., detection wavelength 254 nm) to obtain 37-p1 (33mg, 21%) as a white solid and 37-p2 (102mg, 65%) as a white solid.
Compound 37-p1: LC-MS (ESI) M/z 594.3 (M + H) + 。 1 H NMR(400MHz,DMSO-d 6 )δ7.46–7.37(m,3H),7.32(dt,J＝7.1,1.7Hz,1H),6.97(t,J＝8.0Hz,1H),6.86–6.80(m,2H),6.68(t,J＝9.1Hz,1H),6.25–6.16(m,2H),5.53(t,J＝6.0Hz,1H),5.38(d,J＝8.1Hz,1H),4.94(q,J＝9.0Hz,2H),4.39(d,J＝5.9Hz,2H),3.74(s,2H),3.32–3.25(m,2H),2.95(t,J＝5.8Hz,2H),2.57(t,J＝5.9Hz,2H),2.31–2.21(m,7H),2.13–2.05(m,2H),1.91–1.82(m,2H),1.42–1.25(m,4H).
Compound 37-p2: LC-MS (ESI) M/z 594.3 (M + H) + 。 1 H NMR(400MHz,DMSO-d 6 )δ7.46–7.37(m,3H),7.33(dt,J＝7.0,1.7Hz,1H),7.01–6.91(m,2H),6.84(d,J＝8.2Hz,1H),6.69(t,J＝9.1Hz,1H),6.25–6.18(m,2H),5.55(t,J＝6.0Hz,1H),5.30(d,J＝7.5Hz,1H),4.94(q,J＝9.0Hz,2H),4.39(d,J＝5.8Hz,2H),3.76(s,2H),3.58–3.53(m,2H),2.97(t,J＝5.8Hz,2H),2.59(t,J＝5.9Hz,2H),2.23–2.13(m,7H),1.85–1.74(m,4H),1.68–1.60(m,2H),1.56–1.48(m,2H).
Synthesis of intermediate Compound 38-5
Step 1: synthesis of Compound 38-5a
To the reaction flask was added phthalimide (5.18g, 35.210mmol), DMF (65 mL), K 2 CO 3 (13.27g, 96.027mmol), 3-bromobenzyl bromide (8g, 32.009mmol). The mixture was stirred at room temperature overnight. After the reaction, the reaction solution was added to 300mL of water, stirred for half an hour and then filtered, the obtained solid was dried, then slurried with 30mL PE/EA (10/1) mixed solvent, filtered and dried to obtain the target compound 38-5a (9.41g, 92.99%) as a white solid. 1 H NMR(400MHz,Chloroform-d)δ7.85(dd,J＝5.5,3.1Hz,2H),7.71(dd,J＝5.5,3.1Hz,2H),7.56(t,J＝1.8Hz,1H),7.42–7.33(m,2H),7.18(t,J＝7.8Hz,1H),4.80(s,2H).
And 2, step: synthesis of Compound 38-5b
2-Nitrocarbazole (3.2g, 15.079mmol), compound 38-5a (7.15g, 22.619mmol) and NaHCO are added into a reaction bottle 3 (3.80g, 45.238mmol), cuI (2.87g, 15.079mmol), DMF (35 mL), (1R, 2R) - (-) -N, N' -dimethyl-1, 2-cyclohexanediamine (2.15g, 15.079mmol). After the mixture was further purged with nitrogen 3 to 5 times, it was stirred at 140 ℃ overnight. After the reaction, dichloromethane and water are added for dilution. The organic phase was separated and the aqueous phase was extracted with dichloromethane. The combined organic phases were washed with saturated aqueous NaCl solution, concentrated under reduced pressure, and the crude product was purified by column chromatography (Biotage) (mobile phase: petroleum ether/dichloromethane 100/0 to 0/100), under reduced pressureAfter concentration, the resulting solid was slurried with 100mL of a mixed solvent of petroleum ether/ethyl acetate (10/1), filtered under reduced pressure, and the solid was dried to obtain 38-5b (2.78g, 41.20%) as a pale yellow solid, the objective compound. LC-MS (ESI) M/z 448.1 (M + H) + .
And step 3: synthesis of Compound 38-5
The compound 38-5b (1.8g, 4.02mmol), ethanol (150 mL), hydrazine hydrate (75 mL) were charged into a reaction flask, and after the reaction was completed, the reaction flask was stirred at 80 ℃ for 2 hours, cooled to room temperature, and diluted with ethyl acetate and water. The organic phase was separated and the aqueous phase was extracted with ethyl acetate. Combining the organic phases, washing with saturated aqueous NaCl solution and anhydrous Na 2 SO 4 Drying, filtration and concentration under reduced pressure gave compound 38-5 as a yellow solid (1.2g, 96%). LC-MS (ESI) M/z 318.1 (M + H) + .
EXAMPLE 38 synthetic route to Compound 38-p1 and Compound 38-p2
Step 1: synthesis of Compound 38-1
To a reaction flask was added 8-bromo-5-fluoroisoquinoline (1g, 4.424mmol), acOH (10 mL), naBH 4 (0.59g, 15.484mmol). The mixture was stirred at room temperature for 1 hour. After the reaction, the reaction mixture was concentrated under reduced pressure, and a saturated aqueous sodium bicarbonate solution and methylene chloride were added. The organic phase was separated and the aqueous phase was extracted with dichloromethane. The organic phases were combined, washed with a saturated aqueous NaCl solution, and concentrated under reduced pressure to give the desired compound 38-1 as a colorless oil (1.04 g,>99％)。LC-MS(ESI):m/z 230.0(M+H) + .
step 2: synthesis of Compound 38-2
To a reaction flask was added compound 38-1 (1.01g, 4.390mmol), DCM (15 mL), DIEA EthylisopyllaMine (1.70g, 13.170mmol), BOC 2 O (1.44g, 6.585mmol). The mixture was stirred at 25 ℃ for 1 hour. The reaction mixture was directly concentrated under reduced pressure, and the resulting crude product was purified by an automatic column chromatography (Biotage) (mobile phase: petroleum ether/ethyl acetate 100/0 to 95/5) to obtain the objective compound 38-2 (1.21g, 83.48%) as a colorless oil. LC-MS (ESI) M/z 274.0 (M-tBu) + .
And step 3: synthesis of Compound 38-3
To a reaction flask, compound 38-2 (1.2g, 3.634mmol), DMF (20 mL), sodium thiomethoxide (0.31g, 4.361mmol) was added. The mixture was stirred at 100 ℃ for 1 hour with a microwave. After the reaction, ethyl acetate and water were added to dilute. The organic phase was separated and the aqueous phase was extracted with ethyl acetate. The combined organic phases were washed with saturated aqueous NaCl solution and concentrated under reduced pressure, and the crude product was purified by column chromatography (Biotage) (mobile phase: petroleum ether/ethyl acetate 100/0 to 95/5) to give the title compound 38-3 (569mg, 43.70%) as a colorless oil. LC-MS (ESI): M/z 302.0 (M-tBu) + .
And 4, step 4: synthesis of Compound 38-4
Add compound 38-3 (560mg, 1.5631 mmol), DCM (10 mL), M-CPBA (809.13mg, 4.689mmol) to the flask. The mixture was stirred at 25 ℃ for 2 hours. After the reaction, the reaction mixture was concentrated under reduced pressure, and the resulting concentrate was stirred with saturated sodium bicarbonate for 0.5 hour. Add ethyl acetate and water to dilute. The organic phase was separated and the aqueous phase was extracted with ethyl acetate. The combined organic phases were washed with saturated aqueous NaCl solution and concentrated under reduced pressure, and the crude product was purified by column chromatography (Biotage) (mobile phase: petroleum ether/ethyl acetate 100/0 to 70/30) to give the title compound 38-4 (559mg, 91.64%) as a colorless oil. LC-MS (ESI) M/z 334.0 (M-tBu) + .
And 5: synthesis of Compound 38-6
To the reaction flask were added compound 38-4 (140mg, 0.441mmol), compound 38-5 (258.27mg, 0.662mmol), pd2 (dba) 3 (40.40mg, 0.044mmol), XPHOS (42.06mg, 0.088mmol), cs2CO3 (287.47mg, 0.882mmol), 1,4-dioxane (6 mL). After the mixture was purged with nitrogen 3 to 5 times, it was stirred at 100 ℃ overnight. After the reaction, ethyl acetate and water were added to dilute the reaction mixture. The organic phase was separated and the aqueous phase was extracted with ethyl acetate. The combined organic phases were washed with saturated aqueous NaCl and concentrated under reduced pressure, and the crude product was purified by automatic column chromatography (Biotage) (mobile phase: petroleum ether/ethyl acetate 100/0 to 52/48) to give the title compound 38-6 (223mg, 80.66%) as a pale yellow solid. LC-MS (ESI) of M/z 644.4 (M + NH) 4 ) + .
Step 6: synthesis of Compound 38-7
Add Compound 38-6 (220mg, 0.351mmol), etOH (30 mL), NH to the reaction flask 4 Cl aq. (10 mL), fe (117.60mg, 2.106mmol). The mixture was stirred at 70 ℃ for 1 hour. After the reaction, ethyl acetate and water were added to dilute. The organic phase was separated and the aqueous phase was extracted with ethyl acetate. The combined organic phases were washed with saturated aqueous NaCl solution and concentrated under reduced pressure, and the crude product was purified by automatic column chromatography (Biotage) (mobile phase: petroleum ether/ethyl acetate 100/0 to 50/50) to give the title compound 38-7 as a pale yellow solid (150mg, 71.61%). LC-MS (ESI) M/z 597.3 (M + H) + 。
And 7: synthesis of Compound 38-8
To the reaction flask was added compound 38-7 (145mg, 0.243mmol), etOH (6 mL), THF (2 mL), 4-dimethylaminocyclohexanone (171.56mg, 1.215mmol), ti (OEt) 4 (277.15mg, 1.215mmol). The mixture was stirred at 50 ℃ overnight. The next day, the mixture was cooled to room temperature and NaBH was added 3 CN (152.70mg, 2.430mmol), the mixture is stirred at 80 ℃ for 1 hour. After the reaction, ethyl acetate and water were added to dilute. The organic phase was separated and the aqueous phase was extracted with ethyl acetate. The combined organic phases were washed with saturated aqueous NaCl and concentrated under reduced pressure, and the crude product was purified by automatic column chromatography (Biotage) (mobile phase: dichloromethane/0.5M ammonia in methanol 100/0 to 88/12) to give the title compound 38-8 (136mg, 77.53%) as a yellow oil. LC-MS (ESI) M/z 722.9 (M + H) + 。
And 8: synthesis of Compounds 38-p1 and 38-p2
To the reaction flask was added compound 38-8 (135mg, 0.187mmol), dichloromethane (12 mL), trifluoroacetic acid (4 mL). After the mixture was stirred at room temperature for 1 hour, a saturated aqueous sodium bicarbonate solution was added to neutralize the reaction, and the reaction mixture was diluted with dichloromethane. The organic phase was separated and the aqueous phase was extracted with dichloromethane. The organic phases were combined, washed with saturated aqueous NaCl solution, and concentrated under reduced pressure to purify the crude preparation (Boston pHlex ODS, 21.2X 250mm,10um, water (0.05% trifluoroacetic acid)/acetonitrile, flow rate 30mL/min, column temperature 25 ℃, detection wavelength 254 nm) to obtain 38-p1 (10 mg, 9%) as a white solid and 38-p2 (10 mg, 12%) as a white solid.
Compound 38-p1: LC-MS (ESI) M/z 622.3 (M + H) + 。 1 H NMR(400MHz,DMSO-d 6 )δ7.94–7.88(m,1H),7.83(d,J＝8.5Hz,1H),7.60(t,J＝7.8Hz,1H),7.55(d,J＝8.8Hz,1H),7.50(d,J＝1.8Hz,1H),7.47–7.40(m,2H),7.16–7.10(m,2H),7.09–7.03(m,1H),6.58(dd,J＝8.5,1.9Hz,1H),6.55–6.45(m,3H),5.64(d,J＝7.9Hz,1H),4.57(d,J＝5.9Hz,2H),3.69(s,2H),3.17–3.10(m,1H),3.03(s,3H),3.00–2.94(m,2H),2.88(t,J＝5.6Hz,2H),2.18–2.12(m,7H),2.06–2.00(m,2H),1.85–1.75(m,2H),1.31–1.25(m,2H),1.19–1.08(m,2H).
Compound 38-p2: LC-MS (ESI) M/z 622.3 (M + H) + 。 1 H NMR(400MHz,DMSO-d 6 )δ7.94–7.88(m,1H),7.83(d,J＝8.5Hz,1H),7.60(t,J＝7.7Hz,1H),7.54(d,J＝8.8Hz,1H),7.51(s,1H),7.48–7.40(m,2H),7.15–7.09(m,2H),7.09–7.04(m,1H),6.65(dd,J＝8.5,1.9Hz,1H),6.57–6.47(m,3H),5.73(d,J＝7.5Hz,1H),4.56(d,J＝6.0Hz,2H),3.69(s,2H),3.41–3.38(m,1H),3.03(s,3H),3.00–2.95(m,2H),2.89(t,J＝5.6Hz,2H),2.16(s,6H),2.10–2.04(m,1H),1.74–1.60(m,4H),1.56–1.45(m,4H).
Example 39: synthetic route to compound 39
Step 1: synthesis of Compound 39-1
To the reaction flask was added 2-bromo-1-chloro-4-fluorobenzene (5g, 23.873mmol), DMF (30 mL), sodium thiomethoxide (2.01g, 28.648mmol). The mixture was stirred at 100 ℃ for 1 hour. After the reaction, the reaction solution was added to a saturated aqueous ammonium chloride solution, and then diluted with dichloromethane and water. The organic phase was separated and the aqueous phase was extracted with dichloromethane. The combined organic phases were washed with saturated aqueous NaCl solution and concentrated under reduced pressure to give the title compound 39-1 (8.17 g, > 99%) as a pale yellow liquid, which was used directly in the next reaction.
Step 2: synthesis of Compound 39-2
Add compound 39-1 (5.67g, 23.870mmol) and DCM (80 mL) to the reaction flask, and slowly add m-CPBA (12.36g, 71.609mmol). The mixture was stirred at 25 ℃ for 1 hour. After the reaction is finished, the reaction liquid is decompressed and concentrated to obtainThe concentrate was stirred for 0.5 hour with saturated sodium bicarbonate. Add ethyl acetate and water to dilute. The organic phase was separated and the aqueous phase was extracted with ethyl acetate. The combined organic phases were washed with saturated aqueous NaCl solution and concentrated under reduced pressure, and the crude product was purified by automatic column chromatography (Biotage) (mobile phase: petroleum ether/ethyl acetate 100/0 to 70/30) to give the title compound 39-2 as a white solid (1.76g, 27.36%). 1 H NMR(400MHz,Chloroform-d)δ8.20(d,J＝2.1Hz,1H),7.82(dd,J＝8.3,2.1Hz,1H),7.66(d,J＝8.4Hz,1H),3.07(s,3H).
And 3, step 3: synthesis of Compound 39-3
Add compound 39-2 (600mg, 2.226mmol), zn (CN) to the flask 2 (313.65mg,2.671mmol)，Pd(PPh 3 ) 4 (257.23mg, 0.223mmol), DMF (6 mL). After the mixture was purged with nitrogen 3 to 5 times, it was stirred at 100 ℃ overnight. After the reaction, ethyl acetate and water were added to dilute the reaction mixture. The organic phase was separated and the aqueous phase was extracted with ethyl acetate. The combined organic phases were washed with saturated aqueous NaCl and concentrated under reduced pressure, and the crude product was purified by automatic column chromatography (Biotage) (mobile phase: petroleum ether/ethyl acetate 100/0 to 70/30) to give the title compound 39-3 as a white solid (106mg, 22.08%). 1 H NMR(400MHz,Chloroform-d)δ8.26(d,J＝2.2Hz,1H),8.10(dd,J＝8.5,2.2Hz,1H),7.76(d,J＝8.5Hz,1H),3.10(s,3H).
And 4, step 4: synthesis of Compound 39-4
Intermediate 4-4 (100mg, 0.184mmol), compound 39-3 (47.51mg, 0.220mmol), ruPhos Pd G3 (15.37mg, 0.018mmol), ruPhos (17.14mg, 0.037mmol), cs2CO3 (119.64mg, 0.367mmol), 1,4-dioxane (8 mL) were added to the reaction flask. After the mixture was purged with nitrogen 3 to 5 times, it was stirred at 100 ℃ overnight. After the reaction, ethyl acetate and water were added to dilute. The organic phase was separated and the aqueous phase was extracted with ethyl acetate. The combined organic phases were washed with saturated aqueous NaCl solution, concentrated under reduced pressure and the crude product was purified by column chromatography (Biotage) (mobile phase: dichloromethane/0.5M methanolic ammonia 100/0 to 90/10) to give the title compound 39-4 as a pale yellow solid (103mg, 77.50%). LC-MS (ESI) M/z 725.4 (M + H) + 。
And 5: synthesis of Compound 39-5
Adding the compound into a reaction flask39-4 (100mg, 0.138mmol), meOH (5 mL), 7M NH3-MeOH (1 mL), RANEY NICKEL (100mg, 0.458mmol). The mixture was purged with nitrogen 3 to 5 times, then purged with hydrogen 3 to 5 times, and stirred at 25 ℃ overnight for 1 hour. After the reaction, the reaction mixture was filtered, the filtrate was concentrated under reduced pressure, and the crude product was purified by an automatic column chromatography (Biotage) (mobile phase: dichloromethane/0.5M methanolic ammonia 100/0 to 90/10) to obtain the objective compound 39-5 (70mg, 69.61%) as a pale yellow solid. LC-MS (ESI) M/z 728.9 (M + H) + 。
Step 6: synthesis of Compound 39
To the reaction flask were added 39-5 (70mg, 0.096 mmol), dichloromethane (12 mL), and trifluoroacetic acid (3 mL). After the mixture was stirred at room temperature for 1 hour, a saturated aqueous sodium bicarbonate solution was added to neutralize the reaction, and the reaction mixture was diluted with dichloromethane. The organic phase was separated and the aqueous phase was extracted with dichloromethane. The organic phases were combined, washed with saturated aqueous NaCl, concentrated under reduced pressure and the crude product purified (Boston pHlex ODS,21.2 x 250mm,10um, water (0.05% trifluoroacetic acid)/acetonitrile, flow rate 30mL/min, column temperature 25 ℃, detection wavelength 254 nm) to give target compound 39 (36mg, 59.63%) as a white solid.
Compound 39: LC-MS (ESI): M/z 628.3 (M + H) + 。 1 H NMR(400MHz,DMSO-d 6 )δ7.60(d,J＝2.3Hz,1H),7.52–7.34(m,6H),7.02–6.93(m,2H),6.83(d,J＝8.2Hz,1H),6.59(d,J＝8.6Hz,1H),6.20(d,J＝7.8Hz,1H),5.31(d,J＝7.6Hz,1H),4.98(q,J＝8.9Hz,2H),4.55(d,J＝5.9Hz,2H),3.81(s,2H),3.58–3.49(m,1H),3.01(s,3H),2.17(s,6H),2.08–2.01(m,1H),1.84–1.72(m,4H),1.69–1.59(m,2H),1.54–1.45(m,2H).
Example 40: synthetic route to compound 40
Step 1: synthesis of Compound 40-1
Intermediate B (230mg, 0.68mmol), N-t-butoxycarbonyl-4-piperidone (677mg, 3.4 mmol), tetraethyltitanate (775mg, 3.4 mmol), and ethanol (20 mL) were added to the reaction flask. The mixture was stirred overnight at 50 ℃ under nitrogenSodium cyanoborohydride (214mg, 3.4 mmol) was added and stirring continued at room temperature for 2 hours. The reaction was quenched by addition of saturated aqueous sodium bicarbonate. Filtering, and adding ethyl acetate and water to dilute the filtrate. The organic phase was separated and the aqueous phase was extracted with ethyl acetate. The combined organic phases were washed with saturated aqueous NaCl and concentrated under reduced pressure, and the crude product was purified by autosampler (Biotage) (mobile phase: petroleum ether/ethyl acetate 100/0 to 40/60) to give the title compound 40-1 as a brown solid (300mg, 85%). LC-MS (ESI): M/z 524.1 (M + H) + 。
Step 2 Synthesis of Compound 40-2
Add compound 40-1 (150mg, 0.29mmol), intermediate 1-1 (183mg, 0.44mmol), palladium tetrakistriphenylphosphine (35mg, 0.03mmol), potassium carbonate (80mg, 0.58mmol), N, N-dimethylformamide (5 mL), water (1 mL) to the reaction flask. The nitrogen was replaced 3 to 5 times. The mixture was stirred at 100 ℃ for 2 hours under nitrogen. After the reaction, ethyl acetate and brine were added to dilute the reaction mixture. The organic phase was separated and the aqueous phase was extracted with ethyl acetate. The combined organic phases were washed with saturated aqueous NaCl solution, concentrated under reduced pressure and the crude product was purified by automatic column chromatography (Biotage) (mobile phase: petroleum ether/ethyl acetate 100/0 to 20/80) to give target compound 40-2 as a brown solid (311 mg, crude product). LC-MS (ESI) M/z 687.1 (M + H) + .
And step 3: synthesis of Compound 40-3
To a reaction flask was added compound 40-2 (160 mg, crude), dioxane hydrochloride (4 mL) and stirred at 25 ℃ for one hour. The reaction solvent was distilled off to leave 40-3 (170 mg, crude product) as a solid target compound. LC-MS (ESI) M/z 587.1 (M + H) + .
And 4, step 4: synthesis of Compound 40
To the reaction flask was added compound 40-3 (140 mg, crude), (methoxymethyl) oxirane (110mg, 1.3mmol), potassium carbonate (94mg, 0.68mmol), N, N-dimethylformamide (2 mL). Reaction solution is introduced with N 2 Balloon, stir at 50 ℃ overnight. The next day, the reaction was diluted with ethyl acetate and water. The organic phase was separated and the aqueous phase was extracted with ethyl acetate. The organic phases were combined, washed with saturated NaCl and anhydrous Na 2 SO 4 Drying, filtering and evaporating to obtain a crude product. Purification of the crude preparation obtained (Welch Xt)imate C18, 21.2 × 250mm,10um, water (10 mM ammonium bicarbonate)/acetonitrile, flow rate of 30mL/min, column temperature 25 ℃, detection wavelength 254 nm) to give white solid target compound 40: LC-MS (ESI) M/z 675.1 (M + H) +. 1 H NMR(400MHz,DMSO-d 6 )δ7.50–7.42(2H,m),7.41–7.31(2H,m),7.25–7.16(2H,m),6.97(1H,t,J＝8.0Hz),6.84(2H,d,J＝7.6Hz),6.74(1H,t,J＝6.4Hz),6.54(1H,d,J＝8.4Hz),6.20(1H,d,J＝7.8Hz),5.40(1H,d,J＝8.0Hz),4.97(2H,q,J＝9.0Hz),4.50(3H,d,J＝5.9Hz),3.91(3H,s),3.73(1H,d,J＝5.7Hz),3.32–3.28(2H,m),3.25(3H,s),3.24–3.18(1H,m),3.04(3H,s),2.88(2H,t,J＝13.3Hz),2.34–2.23(2H,m),2.11(2H,q,J＝10.3,9.5Hz),1.94(2H,d,J＝12.3Hz),1.48(2H,dd,J＝16.1,6.6Hz).
Example 41: synthetic routes to Compounds 41-p1 and 41-p2
Step 1: synthesis of Compound 41-1
4- { [4- (dimethylamino) cyclohexyl ] was added to a 50mL three-necked flask in this order]Amino } -2-iodo- (2, 2-trifluoroethyl) indole (576 mg, 1.238mmol), 3-aminomethylphenylboronic acid pinacol ester hydrochloride (400.46mg, 1.485mmol), potassium carbonate (598.82mg, 4.333mmol), tetrakistriphenylphosphine palladium (143.06mg, 0.124mmol), N, N-dimethylformamide (10 mL), and water (2 mL). And reacting for 2 hours at 100 ℃ under the protection of nitrogen. The reaction was run by TLC and water and ethyl acetate were added to the reaction. The organic phase was separated and the aqueous phase was extracted with ethyl acetate. Combining the organic phases, washing with water, washing with saturated NaCl, anhydrous Na 2 SO 4 Drying, filtration, evaporation to dryness, and column purification (mobile phase: dichloromethane/methanol 10/1 to 5/1) to give compound 41-1[ 2- (4- { [4- (dimethylamino) cyclohexyl ]]Amino } -1- (2, 2 trifluoroethyl) indol-2-yl) phenyl]Methylamine (330mg, 0.742mmol, 59.97%). LC-MS (ESI): M/z 445.2 (M + H) + .
Step 2: synthesis of Compound 41-2
To a 50mL three-necked flask, compound 41-1 (330mg, 0.742mmol), 2-methylpropan-2-yl 2-chloro-5, 6,7, 8-tetrahydro-were added in this orderPyrido [4,3-b ]]Pyridine-6-carboxylate (199.49mg, 0.742mmol), 2-dicyclohexanylphosphino-2' - (N, N-dimethylamine) -biphenyl (58.43mg, 0.148mmol), sodium tert-butoxide (107.01mg, 1.113mmol), allylpalladium (II) chloride dimer (27.16mg, 0.074mmol), and toluene (10 mL). And reacting for 18h at 100 ℃ under the protection of nitrogen. The reaction was completed by TLC, and water and ethyl acetate were added to the reaction. The organic phase was separated and the aqueous phase was extracted with ethyl acetate. Combining the organic phases, washing with water, washing with saturated NaCl, anhydrous Na 2 SO 4 Drying, filtering, evaporating to dryness, and purifying by column chromatography (mobile phase: dichloromethane/methanol 15/1 to 10/1) to obtain compound (41-2) 2-methylpropan-2-yl 2- ({ [3- (4- { [4- (dimethylamino) cyclohexyl) amino)]Amino } -1 (2, 2-trifluoroethyl) indol-2-yl) phenyl]Methyl } amino) -5,6,7,8 tetrahydropyrido [4,3-b]Pyridine-6-carboxylate (150mg, 0.222mmol, 29.86%). LC-MS (ESI) M/z 677.5 (M + H) + .
And step 3: synthesis of Compound 41-p1 and Compound 41-p2
To a 50mL three-necked flask were added sequentially compound 41-2 (130mg, 0.192mmol), DCM (10 mL), followed by TFA (2 mL) at 0 ℃. The reaction was carried out at room temperature for 1 hour. The completion of the reaction was checked by TLC, and water and dichloromethane were added to the reaction. The organic phase was separated and the aqueous phase was extracted with dichloromethane. The organic phases were combined, washed with water, saturated NaCl, anhydrous Na 2 SO 4 Drying, filtering, evaporating to dryness, purifying by column (mobile phase: dichloromethane/methanol 10/1 to 5/1), and then preparing compound 41-p1 (25 mg) and compound 41-p2 (25 mg) as white solids by an acid method.
Compound 41-p1: LC-MS (ESI) M/z 577.3 (M + H) + ； 1 H NMR(400MHz,DMSO-d 6 ):δ7.47–7.38(m,2H),7.41–7.33(m,1H),7.37–7.29(m,1H),7.06–6.96(m,1H),6.96(d,J＝8.0Hz,1H),6.84(q,J＝4.6,3.8Hz,3H),6.29(d,J＝8.3Hz,1H),6.19(d,J＝7.7Hz,1H),5.38(d,J＝8.1Hz,1H),4.98(d,J＝9.0Hz,2H),4.50(d,J＝6.1Hz,2H),3.65(s,2H),3.38(s,1H),2.94(t,J＝5.9Hz,2H),2.54(d,J＝5.9Hz,1H),2.18(s,6H),2.08(d,J＝11.7Hz,2H),1.84(d,J＝12.0Hz,2H),1.37–1.21(m,6H).
Compound 41-p1: LC-MS (ESI) M/z 577.3 (M + H) + ； 1 H NMR(400MHz,DMSO-d 6 ):δ7.46-7.40(m,2H),7.36(ddd,J＝14.8,7.6,1.7Hz,2H),7.02(d,J＝8.3Hz,1H),6.99–6.92(m,2H),6.83(dd,J＝7.3,5.1Hz,2H),6.29(d,J＝8.3Hz,1H),6.20(d,J＝7.8Hz,1H),5.32(d,J＝7.5Hz,1H),4.98(q,J＝9.0Hz,2H),4.51(d,J＝6.2Hz,2H),3.64(s,2H),3.53(d,J＝7.5Hz,1H),2.93(t,J＝5.9Hz,2H),2.53(d,J＝5.9Hz,1H),2.17(s,6H),2.05(dt,J＝7.0,3.5Hz,1H),1.89-1.72(m,4H),1.65(dd,J＝8.6,4.1Hz,2H),1.56–1.41(m,3H).
EXAMPLE 42 synthetic route to Compound 42-p1 and Compound 42-p2
Step 1: synthesis of Compound 42-1
To a 50mL three-necked flask were added 4-chloro-5-fluoro-1H-indole (300mg, 1.769mmol) and tetrahydrofuran (10 mL) in that order, followed by sodium hydrogen (106.14mg, 2.654mmol) at 0 deg.C. The reaction was continued at 0 ℃ for 1h. Benzenesulfonyl chloride (624.87mg, 3.538mmol) was then added to the reaction. The reaction was continued at 0 ℃ for 1h. The reaction was completed by TLC, and saturated aqueous ammonium chloride solution and ethyl acetate were added to the reaction. The organic phase was separated and the aqueous phase was extracted with ethyl acetate. The combined organic phases were washed with water, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, evaporated to dryness and purified by column chromatography (mobile phase: petroleum ether/ethyl acetate 25/1 to 20/1) to give 4-chloro-1- (dioxophenyl-. Lambda.6-sulfanyl) -5-fluoroindole (490mg, 1.503mmol, 84.95%). LC-MS (ESI): M/z 310.0 (M + H) + .
Step 2: synthesis of Compound 42-2
To a 50mL three-necked flask, compound 42-1 (487mg, 1.572mmol) and tetrahydrofuran (20 mL) were added in this order, followed by lithium diisopropylamide (2.358 mL) at-70 ℃. The reaction was continued at-78 ℃ for 1h. Elemental iodine (598.59mg, 2.358mmol) was then added to the reaction. The reaction was continued at-78 ℃ for 1h. The completion of the reaction was checked by TLC, and saturated aqueous ammonium chloride solution and ethyl acetate were added to the reaction. The organic phase was separated and the aqueous phase was extracted with ethyl acetate. Mixing organic phases, washing with water, washing with saturated salt solution, drying with anhydrous sodium sulfate, filtering, evaporating,purification by column chromatography (mobile phase: petroleum ether/ethyl acetate 20/1 to 10/1) gave the compound 4-chloro-1- (dioxophenyl-. Lamda.6-sulfanyl) -5-fluoro-2-iodoindole (280mg, 0.643mmol, 40.88%). LC-MS (ESI): M/z 435.8 (M + H) + .
And step 3: synthesis of Compound 42-3
To a 50mL three-necked flask, compound 42-2 (274mg, 0.629mmol), potassium carbonate (260.77mg, 1.887mmol) and methanol (20 mL) were sequentially added, followed by reaction at 70 ℃ for 2h. The reaction was completed by TLC, and water and ethyl acetate were added to the reaction. The organic phase was separated and the aqueous phase was extracted with ethyl acetate. The organic phases were combined, washed with water, saturated NaCl, anhydrous Na 2 SO 4 Drying, filtering, evaporating to dryness and purifying by column chromatography (mobile phase: petroleum ether/ethyl acetate 20/1 to 10/1) to give the compound 4-chloro-5-fluoro-2-iodo-1H-indole (135mg, 0.457mmol, 72.64%). LC-MS (ESI): M/z 295.9 (M + H) + .
And 4, step 4: synthesis of Compound 42-4
To a 50mL three-necked flask, compound 42-3 (133mg, 0.450mmol) and tetrahydrofuran (10 mL) were added in this order, followed by addition of sodium hydrogen (54.01mg, 1.350mmol) at 0 ℃. The reaction was then continued at 0 ℃ for 1h. 2, 2-trifluoroethyl trifluoromethanesulfonate (208.94mg, 0.900mmol) was then added at 0 ℃. The reaction was then continued at 0 ℃ for 1h. The reaction was completed by TLC, and water and ethyl acetate were added to the reaction. The organic phase was separated and the aqueous phase was extracted with ethyl acetate. Combining the organic phases, washing with water, washing with saturated NaCl, anhydrous Na 2 SO 4 Drying, filtering, evaporating to dryness and purifying by column (mobile phase: petroleum ether/ethyl acetate 20/1 to 10/1) to obtain the compound 4-chloro-5-fluoro-2-iodo-1- (2, 2-trifluoroethyl) indole (30mg, 0.079mmol, 17.66%). 91mg of the starting material was recovered. LC-MS (ESI) M/z 377.8 (M + H) + .
And 5: synthesis of Compound 42-5
A50 mL three-necked flask was charged with the compound 42-4 (150mg, 0.3972 mmol), [ 2-methoxy-4- (methyldioxy-. Lambda.6-sulfanyl) phenyl group]{ [3- (4, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) phenyl]Methyl } amine (248.73mg, 0.596mmol), potassium carbonate (109.82mg, 0.795 mmol), tetrakistriphenylphosphine palladium (45.92mg, 0.040mmol), N, N-dimethylformamide(10 mL) and water (2 mL). The reaction was carried out at 100 ℃ for 2h. The reaction was run by TLC and water and ethyl acetate were added to the reaction. The organic phase was separated and the aqueous phase was extracted with ethyl acetate. The organic phases were combined, washed with water, saturated NaCl, anhydrous Na 2 SO 4 Drying, filtering, evaporating to dryness, and purifying with column (mobile phase: dichloromethane/methanol 20/1 to 15/1) to obtain 4-chloro-5-fluoro-2- [3- ({ [ 2-methoxy-4- (methyldioxo-lambda 6-sulfanyl) phenyl) compound]Amino } methyl) phenyl]-1- (2, 2-trifluoroethyl) indole (140mg, 0.259mmol, 65.13%). LC-MS (ESI) M/z 541.1 (M + H) + .
Step 6: synthesis of Compound 42-p1 and Compound 42-p2
Into a 50mL three-necked flask were charged, in order, compound 42-5 (100mg, 0.185mmol), 4- (dimethylamino) cyclohex-1-amine (105.18mg, 0.739mmol), sodium tert-butoxide (26.65mg, 0.277mmol), 2- (di-tert-butylphosphino) biphenyl (11.03mg, 0.037mmol), tris [ dibenzylideneacetone]Dipalladium (16.93mg, 0.018mmol), toluene (5 mL). The reaction is carried out for 18h at 110 ℃. The reaction was not completely completed by TLC detection, and water and ethyl acetate were added to the reaction. The organic phase was separated and the aqueous phase was extracted with ethyl acetate. The organic phases were combined, washed with water, saturated NaCl, anhydrous Na 2 SO 4 Drying, filtering, evaporating to dryness, purifying by column (mobile phase: dichloromethane/methanol 101 to 5/1), and then preparing compound 42-p1 (37 mg) and compound 42-p2 (25 mg) as white solids by an acid method.
Compound 42-p1: LC-MS (ESI): M/z 647.3 (M + H) + ； 1 H NMR(400MHz,DMSO-d 6 ):δ7.49–7.33(m,4H),7.22(dd,J＝8.3,2.0Hz,1H),7.19(d,J＝2.0Hz,1H),6.95(d,J＝10.7Hz,2H),6.85(s,1H),6.74(t,J＝6.4Hz,1H),6.55(d,J＝8.4Hz,1H),5.01(q,J＝9.0Hz,2H),4.62(dd,J＝9.9,2.9Hz,1H),4.51(d,J＝6.4Hz,2H),3.91(s,3H),3.49(s,1H),3.04(s,3H),2.15(s,7H),1.97(d,J＝8.8Hz,2H),1.78(d,J＝9.5Hz,2H),1.39–1.22(m,4H).
Compound 42-p2: LC-MS (ESI) M/z 647.3 (M + H) + ； 1 H NMR(400MHz,DMSO-d 6 ):δ7.50–7.35(m,4H),7.22(dd,J＝8.3,2.1Hz,1H),7.20(d,J＝2.0Hz,1H),7.07–6.91(m,2H),6.87(s,1H),6.74(t,J＝6.4Hz,1H),6.55(d,J＝8.3Hz,1H),5.02(q,J＝9.0Hz,2H),4.60(dd,J＝9.5,2.9Hz,1H),4.51(d,J＝6.4Hz,2H),3.91(s,3H),3.75(s,1H),3.05(s,3H),2.15(s,6H),2.01(d,J＝4.0Hz,1H),1.80–1.64(m,4H),1.59(dd,J＝8.6,4.0Hz,2H),1.44(dd,J＝12.6,8.9Hz,2H).
Synthetic route of intermediate compound 43-2a
Step 1: synthesis of Compound 43-1a
Into the reaction flask were charged N-Boc-8-bromo-1, 2,3, 4-tetrahydroisoquinoline (5.00g, 16.02mmol), tert-butyl carbamate (3.75g, 32.03mmol), palladium acetate (180mg, 0.8mmol), XPHOS (0.76g, 1.601mmol), cesium carbonate (15.65g, 9.22mmol), 1,4-dioxane (50 mL). N for the mixture 2 The mixture was replaced and stirred at 100 ℃ for 2 hours. The reaction solution was cooled to room temperature and diluted with ethyl acetate and water. The organic phase was separated and the aqueous phase was extracted with ethyl acetate. Combining the organic phases, washing with water, washing with saturated NaCl, anhydrous Na 2 SO 4 Drying, filtration, evaporation to dryness and purification by column chromatography (mobile phase: ethyl acetate/petroleum ether 5% to 20%) gave compound 43-1a (5.25g, 75%). LC-MS (ESI) M/z 349.2 (M + H) + ；1H NMR(400MHz,CDCl 3 ):δ7.65–7.57(m,1H),7.22–7.12(m,1H),6.92(s,1H),6.12(s,1H),4.47(s,2H),3.62(t,J＝5.9Hz,2H),2.83(t,J＝5.6Hz,2H),1.51(s,9H),1.50(s,9H).
And 2, step: synthesis of Compound 43-2a
43-1a (410mg, 1.18mmol), 3-bromo-5-cyanobenzyl bromide (308mg, 1.12mmol), cesium carbonate (864mg, 2.65mmol), and DMF (5 mL) were added to the reaction flask. The mixture was stirred at 50 ℃ for 2 hours. The reaction solution was cooled to room temperature and diluted with ethyl acetate and water. The organic phase was separated and the aqueous phase was extracted with ethyl acetate. Combining the organic phases, washing with water, washing with saturated NaCl, anhydrous Na 2 SO 4 Drying, filtration and evaporation to dryness gave crude product 43-2a (770 mg, 99%). LC-MS (ESI) M/z 544.1 (M + H) +.
Example 43: synthetic route to compound 43
Step 1: synthesis of Compound 43-1
Add intermediate B (620mg, 1.33mmol), pd to the reaction flask 2 (dba) 3 (122mg, 0.13mmol), S-Phos (109mg, 0.27mmol), potassium acetate (392mg, 4.00mmol), toluene (20 mL) in N 2 Stirring was carried out at room temperature for 10 minutes under protection, then pinacolborane (1.19g, 9.33mmol) was added and the mixture was stirred with N 2 The mixture was replaced and stirred at 60 ℃ for 3 hours. The reaction solution was cooled to room temperature, evaporated to dryness and purified by column chromatography (mobile phase: 0.7M methanolic ammonia/dichloromethane 5% to 10%) to give compound 43-1 (183mg, 20%). LC-MS (ESI) M/z 466.0 (M + H) +.
Step 2: synthesis of Compound 43-2
To the reaction flask were added 43-1 (182mg, 0.27mmol), compound 43-2a (293mg, 0.54mmol), tetrakis (triphenylphosphine) palladium (31mg, 27. Mu. Mol), potassium carbonate (112mg, 0.81mmol), DMF (50 mL) and water (1 mL). N for the mixture 2 The mixture was replaced and stirred at 100 ℃ for 2 hours. The reaction solution was cooled to room temperature and diluted with ethyl acetate and water. The organic phase was separated and the aqueous phase was extracted with ethyl acetate. Combining the organic phases, washing with water, washing with saturated NaCl, anhydrous Na 2 SO 4 Drying, filtration, evaporation to dryness and purification on column (mobile phase: 0.7M methanolic ammonia/dichloromethane 5% to 10%) gave compound 43-2 (64mg, 28%). LC-MS (ESI) M/z 802.0 (M + H) +.
And 3, step 3: synthesis of Compound 43
To the reaction flask was added compound 43-2 (64mg, 76. Mu. Mol), DCM (5 mL), TFA (1 mL). After the mixture was stirred at room temperature for 1 hour, a saturated aqueous sodium bicarbonate solution was added to neutralize the reaction, and the reaction mixture was diluted with dichloromethane. The organic phase was separated and the aqueous phase was extracted with dichloromethane. The organic phases were combined, washed with saturated aqueous NaCl solution, and concentrated under reduced pressure to afford crude product purification (Boston pHlex ODS, 21.2X 250mm,10um, water (0.05% trifluoroacetic acid)/acetonitrile, flow rate 30mL/min, column temperature 25 ℃, detection wavelength 254 nm) to afford title compound 43 (35mg, 76%) as a white solid. LC-MS (ESI): M/z 601.3 (M + H) + .1H NMR(400MHz,DMSO-d6)：δ7.87–7.76(m,3H),7.08(s,1H),7.00(t,J＝8.0Hz,1H),6.88(d,J＝8.2Hz,1H),6.81(t,J＝7.8Hz,1H),6.33(d,J＝7.5Hz,1H),6.22(t,J＝7.2Hz,2H),5.70(s,1H),5.35(d,J＝7.5Hz,1H),5.03(q,J＝8.8Hz,2H),4.46(d,J＝5.8Hz,2H),3.75(s,2H),3.62–3.46(m,2H),2.93(t,J＝5.5Hz,2H),2.69–2.59(m,2H),2.18(s,6H),2.10–2.02(m,1H),1.87–1.70(m,4H),1.69–1.58(m,2H),1.56–1.43(m,2H).
Example 44: synthetic route to compound 44
Step 1: synthesis of Compound 44-1
To the reaction flask were added intermediate 4-4 (130mg, 0.24mmol), 2-bromo-5-fluorobenzonitrile (71mg, 0.36mmol), palladium chloride (. Pi. -cinnamyl) dimer (6 mg,12. Mu. Mol), t-Bu-BippyPhos (12mg, 24. Mu. Mol), cesium carbonate (155mg, 0.48mmol), 1,4-dioxane (2 mL). N for the mixture 2 The mixture was replaced and stirred at 100 ℃ for 20 hours. The reaction solution was cooled to room temperature, evaporated to dryness and purified by column chromatography (mobile phase: 0.7M methanolic ammonia/dichloromethane 0% to 10%) to give compound 44-1 (68mg, 42%). LC-MS (ESI) M/z 664.3 (M + H) + .
Step 2: synthesis of Compound 44-2
To a solution of compound 44-1 (68mg, 0.10 mmol) in tetrahydrofuran (5 mL) were added 28% aqueous ammonia (0.3 mL,0.10 mmol) and Raney nickel (100mg, 0.46mmol), and the mixture was washed with H 2 The mixture was replaced and stirred at room temperature for 1 hour. The reaction mixture was filtered and evaporated to dryness to give compound 44-2 (60mg, 67%). LC-MS (ESI): M/z 668.5 (M + H) + .
And 3, step 3: synthesis of Compound 44
Add compound 44-2 (60mg, 0.07mmol), DCM (2 mL), TFA (0.5 mL) to the reaction flask. After the mixture was stirred at room temperature for 1 hour, a saturated aqueous sodium bicarbonate solution was added to neutralize the reaction, and the reaction mixture was diluted with dichloromethane. The organic phase was separated and the aqueous phase was extracted with dichloromethane. The combined organic phases were washed with saturated aqueous NaCl solution and concentrated under reduced pressure to give a crude product which was purified (Boston pHlex ODS, 21.2X 250mm,10um, water (0.05% trifluoroacetic acid)Acetonitrile/acetonitrile flow rate 30mL/min, column temperature 25 ℃, detection wavelength 254 nm) to give target compound 44 (25mg, 63%) as a white solid. LC-MS (ESI): M/z 568.3 (M + H) + .1H NMR(400MHz,DMSO-d6)：δ7.50–7.39(m,3H),7.35(d,J＝6.9Hz,1H),6.96(dd,J＝9.8,6.1Hz,3H),6.82(d,J＝8.2Hz,1H),6.77(td,J＝8.7,3.0Hz,1H),6.41(dd,J＝8.8,4.8Hz,1H),6.35(t,J＝5.5Hz,1H),6.20(d,J＝7.7Hz,1H),5.30(d,J＝7.5Hz,1H),4.94(q,J＝8.7Hz,2H),4.41(d,J＝5.0Hz,2H),3.75(s,2H),3.53(s,1H),2.17(s,6H),2.11–2.02(m,1H),1.86–1.71(m,4H),1.69–1.58(m,2H),1.54–1.42(m,2H).
Biological activity assay
The method comprises the following steps: the CTG method detects the inhibition effect of the candidate compound on the proliferation capacity of 5 cell lines.
Reagents and materials:
Cell Titer-Glo luminescent cell viability assay(Promega,Cat.No.G7573,Lot.No.0000453220)
FBS (from Biological Industries, cat.No.04-002-1A, lot.No. 1841929)
Penicilin-Streptomyces solution (from Invitrogen, cat. No.15140-122, lot. No. 2257222)
RPMI 1640 (from Invitrogen, cat.No.11875-093, lot.No. 2274537)
DMEM (Low glucose) (available from Invitrogen, cat.No.11885-084, lot.No. 2217385)
F-12K (purchased from Invitrogen, cat.No.21127-022, lot.No. 2239709)
0.25% Trypsine-EDTA (from Invitrogen, cat. No.25200-072, lot. No. 2276965)
Dimethyl sulfoxide (Sigma, cat. No.276855-1L, lot. No. SHBL5610)
96-well plates (available from Corning, cat. No. CLS3903; lot. No. 36520038)
Examples biological activity assays:
the CTG method is used for detecting the proliferation inhibition experiment of the NUGC3 cell line in the embodiment of the application; nucc 3 was a P53Y 220C-specific mutant cell line, and the inhibitory effect of examples on P53Y 220C-specific mutant tumor cells was evaluated by examining the proliferation inhibitory activity of examples on nucc 3.
The specific operation method of the experiment is as follows:
1. cell plating
1) Preparing complete culture medium, and mixing completely.
2) Recovering the cells, and selecting cell strains with good growth state after two generations.
3) The cell culture flask was removed from the incubator and the cell name and media type marked on the flask were checked.
4) Adherent cells: the medium was aspirated off, washed once with pancreatin, the waste liquid was discarded, and 3mL of fresh pancreatin was added to the flask for digestion. When the cells loose and are about to separate from the bottle wall, 9mL of complete medium is added to stop the pancreatin digestion, and the mixture is gently mixed. The cell suspension was pipetted into a centrifuge tube and centrifuged at 1000rpm for 5 minutes.
5) The supernatant was discarded.
6) Add the appropriate volume of medium to the centrifuge tube and gently blow it to resuspend the cells evenly.
7) Counting was performed using a Vi-Cell XR cytometer.
8) The cell suspension was adjusted to the appropriate concentration.
9) The cell suspension was added to a 96-well plate at 100. Mu.L/well. Marking cell name, plate density, date and other detailed information, and placing the culture plate in CO 2 The incubator was overnight.
2. Preparation and addition of compound plates:
1) Preparing a compound to be tested:
compounds were formulated in DMSO as 10mM stock solutions. Diluted to 4mM working concentration in DMSO.
2) Staurosporine compound plate formulation:
staurosporine was formulated in DMSO as a 2mM stock. Diluted to 0.4mM working concentration in DMSO.
3. Incubation for 72 hours in a carbon dioxide incubator for reagent preparation and detection
1) CellTiter-Glo Buffer was thawed at room temperature. The lyophilized CellTiter Glo substrate was equilibrated to room temperature.
2) CellTiter-Glo Buffer was added to CellTiter Glo substrate and mixed well.
3) The cell plate was removed and equilibrated to room temperature.
4) Add 100. Mu.L of the mixed CellTiter Glo reagent to each well, shake for 10min in the dark, incubate for 10min.
5) Placing the culture plate into an EnVision reading plate, and recording the luminousnce reading result; the inhibition rate was calculated according to the following formula: inhibition (%) = (1- (RLU test-RLU blank)/(RLU solvent-RLU blank)) × 100%.
6) XLFit was used to plot drug inhibition curves and calculate IC 50 The value is obtained. Using ase:Sub>A 4-parameter model [ fit = (A + ((B-A)/(1 + ((C/x) ^ D))))].
Result + represents IC 50 <=1 μ M + + represents 1<IC 50 <＝10μM+++IC 50 >10μM
N.d. indicates not tested.
Claims (14)
1. A compound of formula I, or an isotopic derivative thereof, or a pharmaceutically acceptable salt of any of the foregoing, or a solvate of any of the foregoing:
wherein ring D is
(# and # are used to identify the connection location);
ring D 1 And ring D 2 In each A 1 、A 2 、A 3 And A 4 Each independently is N or CH;
ring D 3 In (B) 1 、B 3 And B 4 Each independently CH, N, NH, O or S; b is 2 And B 5 Each independently is C or N;
each R a Independently F, cl, br, I, -CN, C 1 -C 4 Alkyl radical, C 1 -C 4 Alkoxy or halo C 1 -C 4 An alkyl group;
m is
Z 1 Is C (R) 3 ) N, O or N (R) 4 )；
Z 2 Is C (R) 5 ) N, O or N (R) 6 )；
Z 3 Is CH or N;
Z 4 is CH or N;
each L 2 Independently of one another is-NH-, -CH 2 -, -O-or-S-;
ring Q 1 And ring Q 2 Each independently is a saturated or partially unsaturated 5-to 10-membered carbocyclic ring, a saturated or partially unsaturated 5-to 10-membered heterocyclic ring, a 6-to 10-membered aromatic ring, a 5-to 10-membered heteroaromatic ring, or an 8-to 10-membered bicyclic fused ring; one ring in the 8-10 membered bicyclic fused ring is a saturated or partially unsaturated 5-6 membered carbocyclic ring or a saturated or partially unsaturated 5-6 membered heterocyclic ring, and the other ring is a benzene ring or a 5-6 membered heteroaromatic ring;
ring D 3 And in ring C, the- -bond is a double bond or a single bond, provided that ring D 3 And ring C is an aromatic ring;
R 3 、R 4 、R 5 and R 6 Each independently is H, C 1 -C 4 Alkyl or halo C 1 -C 4 An alkyl group;
R A and R B Independently is H, halogen or C 1 -C 4 An alkyl group;
L 1 is absent or-C (O) -;
ring B is a saturated or partially unsaturated 3-to 10-membered carbocyclic ring, a saturated or partially unsaturated 3-to 10-membered heterocyclic ring, a 6-to 10-membered aromatic ring, a 5-to 10-membered heteroaromatic ring, or a 6-to 10-membered bicyclic fused ring; one ring in the 6-10 membered bicyclic fused ring is a saturated or partially unsaturated 5-6 membered carbocyclic ring or a saturated or partially unsaturated 5-6 membered heterocyclic ring, and the other ring is a benzene ring or a 5-6 membered heteroaromatic ring;
each R 2 Independently F, cl, br, I, = O, -CN, -NH 2 、-NHR d 、-NR d R e 、C 1 -C 4 Alkyl, by 1,2 or 3R c Substituted C 1 -C 4 Alkyl radical, C 1 -C 4 Alkoxy, with 1,2 or 3R c Substituted C 1 -C 4 Alkoxy, halo C 1 -C 4 Alkyl, -C (O) NHR d 、-C(O)NR d R e 、-C(O)OR d or-SO 2 R d ；
Each R 1 Independently is F, cl, br, I, -NH 2 、＝O、-NHR d 、-NR d R e 、C 1 -C 4 Alkyl, by 1,2 or 3R h Substituted C 1 -C 4 Alkyl radical, C 1 -C 4 Alkoxy, by 1,2 or 3R h Substituted C 1 -C 4 Alkoxy, halo C 1 -C 4 Alkyl, 3-10 membered cycloalkyl or 3-10 membered heterocycloalkyl (e.g.
)；
Each R d And R e Each independently is H, C 1 -C 4 Alkyl or by 1,2 or 3R b Substituted C 1 -C 4 An alkyl group;
each R c 、R b And R h Each independently is-OH, -OR f 、-NH 2 、-NHR f or-NR f R g ；
Each R f And R g Each independently is C 1 -C 4 Alkyl or-Boc;
n is 0,1, 2,3 or 4;
m is 0,1, 2,3 or 4;
p is 0,1, 2,3 or 4;
the number of heteroatoms in the heterocycle, heterocycloalkyl, heteroaryl ring and heteroaryl group is 1,2 or 3, and each heteroatom is independently N, O or S.
2. The compound, isotopic derivative, pharmaceutically acceptable salt or solvate of claim 1, wherein ring D is
(# and # are used to identify the connection location);
ring D 1 And ring D 2 In each A 1 、A 2 、A 3 And A 4 Each independently is N or CH;
ring D 3 In (B) 1 、B 3 And B 4 Each independently CH, N, NH, O or S; b 2 And B 5 Each independently is C or N;
each R a Independently F or-CN;
m is
Z 1 Is N (R) 4 )；
Z 2 Is C (R) 5 )；
Z 3 Is N;
Z 4 is CH;
each L 2 Independently is-NH-;
ring Q 1 And ring Q 2 Each independently is a saturated or partially unsaturated 5-10 membered carbocyclic ring, a saturated or partially unsaturated 5-10 membered heterocyclic ring, a 6-10 membered aromatic ring, a 5-10 membered heteroaromatic ring, or an 8-10 membered bicyclic fused ring; one ring in the 8-10 membered bicyclic fused ring is a saturated or partially unsaturated 5-6 membered carbocyclic ring or a saturated or partially unsaturated 5-6 membered heterocyclic ring, and the other ring is a benzene ring or a 5-6 membered heteroaromatic ring;
ring D 3 And in ring C, the- -bond is a double bond or a single bond, provided that ring D 3 And ring C is an aromatic ring;
R 4 and R 5 Each independently is H or halo C 1 -C 4 An alkyl group;
R A is H or C 1 -C 4 An alkyl group;
R B is H or halogen;
L 1 is absent or-C (O) -;
ring B is a saturated or partially unsaturated 3-to 10-membered carbocyclic ring, a saturated or partially unsaturated 3-to 10-membered heterocyclic ring, a 6-to 10-membered aromatic ring, a 5-to 10-membered heteroaromatic ring, or a 6-to 10-membered bicyclic fused ring; one ring in the 6-10 membered bicyclic fused ring is a saturated or partially unsaturated 5-6 membered carbocyclic ring or a saturated or partially unsaturated 5-6 membered heterocyclic ring, and the other ring is a benzene ring or a 5-6 membered heteroaromatic ring;
each R 2 Independently F, = O, -CN, -NH 2 、-NHR d 、C 1 -C 4 Alkyl, by 1,2 or 3R c Substituted C 1 -C 4 Alkyl radical, C 1 -C 4 Alkoxy, halo C 1 -C 4 Alkyl, -C (O) NHR d or-SO 2 R d ；
Each R 1 Independently F, = O, -NR d R e 、C 1 -C 4 Alkyl, by 1,2 or 3R h Substituted C 1 -C 4 Alkyl or 3-10 membered heterocycloalkyl;
each R d And R e Each independently is H or C 1 -C 4 An alkyl group;
each R c 、R b And R h Each independently is-OH, -OR f 、-NH 2 、-NHR f or-NR f R g ；
Each R f And R g Each independently is C 1 -C 4 Alkyl or-Boc;
n is 0,1 or 2;
m is 0,1 or 2;
p is 0 or 1;
the number of heteroatoms in the heterocycle, heterocycloalkyl, heteroaryl ring and heteroaryl group is 1,2 or 3, and each heteroatom is independently N, O or S.
3. The compound, isotopic derivative, pharmaceutically acceptable salt or solvate of claim 1 or 2, wherein at R A 、R B 、R a 、R 3 、R 4 、R 5 、R 6 、R 2 、R 1 、R d 、R e 、R f And R g In the definition of (1), said C 1 -C 4 Alkyl is independently at each occurrence methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, or tert-butyl;
and/or, at R B 、R a 、R 3 、R 4 、R 5 、R 6 、R 2 And R 1 In the definition of (1), said halogeno C 1 -C 4 C in alkyl 1 -C 4 Alkyl is independently methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl or tert-butyl;
and/or, at R a 、R 3 、R 4 、R 5 、R 6 、R 2 And R 1 In the definition of (1), said halogeno C 1 -C 4 Halo in the alkyl is independently fluoro;
and/or at R a 、R 2 And R 1 In the definition of (1), said C 1 -C 4 Alkoxy is independently at each occurrence methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, isobutoxy, sec-butoxy, or tert-butoxy;
and/or in ring Q 1 And ring Q 2 The 5-10 membered carbocyclic ring is independently a 5,6,7,8, 9 or 10 membered carbocyclic ring;
and/or in ring Q 1 And ring Q 2 In the definition of (1), the 5-10 membered heterocyclic ring is independently a 5,6,7,8, 9 or 10 membered heterocyclic ring;
and/or in ring Q 1 And ring Q 2 In the definition of (1), the 6-to 10-membered aromatic ring is independently a benzene ring or a naphthalene ring;
and/or in ring Q 1 And ring Q 2 In the definition of (1), the 5-10 membered heteroaromatic ring is independently a 5,6,7,8, 9 or 10 membered heteroaromatic ring;
and/or in ring Q 1 And ring Q 2 The 8-10 membered bicyclic fused ring is independently an 8, 9 or 10 membered bicyclic fused ring;
and/or, in the definition of ring B, the 3-10 membered carbocyclic ring is a 3,4, 5,6,7,8, 9 or 10 membered carbocyclic ring;
and/or, in the definition of ring B, the 3-10 membered heterocyclic ring is a 3,4, 5,6,7,8, 9 or 10 membered heterocyclic ring;
and/or, in the definition of ring B, the 6-10 membered aromatic ring is a benzene ring or a naphthalene ring;
and/or, in the definition of ring B, the 5-10 membered heteroaromatic ring is a 5,6,7,8, 9 or 10 membered heteroaromatic ring;
and/or, in the definition of ring B, the 6-10 membered bicyclic fused ring is a 6,7,8, 9 or 10 membered bicyclic fused ring;
and/or, at R 1 In the definition of (1), the 3-to 10-membered cycloalkyl group is a 3,4, 5,6,7,8, 9 or 10-membered cycloalkyl group;
and/or at R 1 In the definition of (1), the 3-10 membered heterocycloalkyl is a 3,4, 5,6,7,8, 9 or 10 membered heterocycloalkyl;
and/or, ring Q 1 And ring Q 2 Carbon atom of (1) with L 2 And (4) connecting.
4. The compound, isotopic derivative, pharmaceutically acceptable salt or solvate of claim 1 or 2, wherein ring D is 1 Is composed of
And/or, ring D 2 Is composed of
And/or, ring D 3 Is composed of
And/or, Z 1 Is C (R) 3 ) Or N (R) 4 )；
And/or, Z 3 Is N;
and/or, R 3 Is halo C 1 -C 4 An alkyl group;
and/or, R 4 Is halo C 1 -C 4 An alkyl group;
and/or, R 5 Is H;
and/or, R 6 Is H;
and/or, p is 0 or 1;
and/or, R a Is F or-CN;
and/or, each L 2 Independently is-NH-;
and/or, ring Q 1 And Q 2 Each independently is
And/or, each R d And R e Each independently is C 1 -C 4 An alkyl group;
and/or, ring B is
5. The compound, isotopic derivative, pharmaceutically acceptable salt or solvate of any one of claims 1 or 2, wherein ring D is
/>
And/or the presence of a gas in the atmosphere,
is structurally characterized as->
E.g. based on->
And/or, R 3 Is fluoro C 2 An alkyl group;
and/or, R 4 Is fluoro C 2 An alkyl group;
and/or, R a Is F or-CN;
and/or, ring Q 1 And Q 2 Each independently is
And/or, each R d And R e Each independently is methyl;
and/or the presence of a gas in the gas,
is->
6. The compound, isotopic derivative, pharmaceutically acceptable salt or solvate of any one of claims 1 to 5,
And/or the presence of a gas in the gas,
is structurally characterized as>
And/or, R 3 is-CH 2 CF 3 ；
And/or, R 4 is-CH 2 CF 3 ；
And/or the presence of a gas in the atmosphere,
each independently is
m is 0,1 or 2 i Is R 1 ，R j Is H or R 1 ；
And/or the presence of a gas in the gas,
is->
/>
And/or, each R 1 Independently F, = O, CH 3 、
And/or, each R 2 Independently F, -NH 2 、-CH 3 、-NHCH 3 、-SO 2 CH 3 、-OCH 3 、-CF 3 、-C(O)NH 2 、＝O、-CN、-CH 2 NHBoc、-CH 2 NH 2 or-C (O) NHCH 3 ；
And/or n is 0,1 or 2.
7. The compound, isotopic derivative, pharmaceutically acceptable salt or solvate of any one of claims 1 to 6,
each independently is
And/or the presence of a gas in the gas,
is->
/>
8. The compound, isotopic derivative, pharmaceutically acceptable salt or solvate of claim 1, wherein said compound has any one of the following structures:
wherein R is A 、R B 、R 1 、m、R 4 、R 5 Ring Q 1 Ring Q 2 、L 2 、R a P, ring B, R 2 And n is as defined in claim 1.
9. The compound, isotopic derivative, pharmaceutically acceptable salt or solvate of claim 1, wherein the compound has any one of the following structures:
10. the compound, isotopic derivative, pharmaceutically acceptable salt or solvate of claim 9, wherein the compound is any one of the following stereoisomers:
11. a pharmaceutical composition comprising a compound, isotopic derivative, pharmaceutically acceptable salt or solvate of any one of claims 1-10, and at least one pharmaceutically acceptable adjuvant.
12. Use of a compound, isotopic derivative, pharmaceutically acceptable salt or solvate of any one of claims 1 to 10, or a pharmaceutical composition of claim 11, in the manufacture of a medicament for use in the treatment of a patient suffering from a neoplasm which carries a p 53Y 220C mutation.
13. The use of claim 12, wherein the tumor is gastric cancer or liver cancer.
14. The compound, isotopic derivative, pharmaceutically acceptable salt or solvate of claim 1, wherein formula I has the structure according to formula II:
wherein ring D is
Ring D 1 And ring D 2 In each A 1 、A 2 、A 3 And A 4 Each independently is N or CH;
ring D 3 In (B) 1 、B 3 And B 4 Each independently CH, N, NH, O or S; b is 2 And B 5 Each independently is C or N;
each R a Independently F, cl, br, I, C 1 -C 4 Alkyl radical, C 1 -C 4 Alkoxy or halo C 1 -C 4 An alkyl group;
m is
/>
Z 1 Is C (R) 3 ) N, O or N (R) 4 )；
Z 2 Is C (R) 5 ) N, O or N (R) 6 )；
Z 3 Is CH or N;
Z 4 is CH or N;
each L 2 Independently of one another is-NH-, -CH 2 -, -O-or-S-;
ring Q 1 And ring Q 2 Each independently is a saturated or partially unsaturated 5-to 10-membered carbocyclic ring, a saturated or partially unsaturated 5-to 10-membered heterocyclic ring, a 6-to 10-membered aromatic ring, a 5-to 10-membered heteroaromatic ring, or an 8-to 10-membered bicyclic fused ring; one ring of the 8-10 membered bicyclic ring is a saturated or partially unsaturated 5-6 membered carbocyclic ring, or a saturated or partially unsaturated 5-6 membered heterocyclic ring, and the other ring isIs a phenyl ring or a 5-6 membered heteroaromatic ring;
ring D 3 And in ring C, the- -bond is a double bond or a single bond, provided that ring D 3 And ring C is an aromatic ring;
R 3 、R 4 、R 5 and R 6 Each independently is H, C 1 -C 4 Alkyl or halo C 1 -C 4 An alkyl group;
L 1 is absent or-C (O) -;
ring B is a saturated or partially unsaturated 3-to 10-membered carbocyclic ring, a saturated or partially unsaturated 3-to 10-membered heterocyclic ring, a 6-to 10-membered aromatic ring, a 5-to 10-membered heteroaromatic ring, or a 6-to 10-membered bicyclic fused ring; one ring in the 6-10 membered bicyclic fused ring is a saturated or partially unsaturated 5-6 membered carbocyclic ring or a saturated or partially unsaturated 5-6 membered heterocyclic ring, and the other ring is a benzene ring or a 5-6 membered heteroaromatic ring;
each R 2 Independently is F, cl, br, I, -NH 2 、-NHR d 、-NR d R e 、C 1 -C 4 Alkyl, by 1,2 or 3R c Substituted C 1 -C 4 Alkyl radical, C 1 -C 4 Alkoxy, by 1,2 or 3R c Substituted C 1 -C 4 Alkoxy, halo C 1 -C 4 Alkyl, -C (O) NHR d 、-C(O)NR d R e 、-C(O)OR d or-SO 2 R d ；
Each R 1 Independently F, cl, br, I, -NH 2 、＝O、-NHR d 、-NR d R e 、C 1 -C 4 Alkyl, by 1,2 or 3R h Substituted C 1 -C 4 Alkyl radical, C 1 -C 4 Alkoxy, with 1,2 or 3R h Substituted C 1 -C 4 Alkoxy, halo C 1 -C 4 Alkyl, 3-10 membered cycloalkyl or 3-10 membered heterocycloalkyl;
each R d And R e Each independently is C 1 -C 4 Alkyl or by 1,2 or 3R b Substituted C 1 -C 4 An alkyl group;
each R c 、R b And R h Each independently is-OH, -OR f 、-NH 2 、-NHR f or-NR f R g ；
Each R f And R g Each independently is C 1 -C 4 An alkyl group;
n is 0,1, 2,3 or 4;
m is 0,1, 2,3 or 4;
p is 0,1, 2,3 or 4;
the number of heteroatoms in the heterocycle, heterocycloalkyl, heteroaryl ring and heteroaryl group is 1,2 or 3, and each heteroatom is independently N, O or S.
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