WO2022171143A1 - 5,6,7,8-tetrahydropyridine[3,4-d] pyrimidine compound - Google Patents

Abstract

A class of 5,6,7,8-tetrahydropyridine [3,4-d] pyrimidine compounds, specifically a compound as shown in formula (II) or a pharmaceutically acceptable salt thereof.

Description

5,6,7,8-Tetrahydropyridine[3,4-d]pyrimidine compounds

The present invention claims the following priority

CN202110182356.1, application date: February 9, 2021;

CN202110252391.6, application date: March 08, 2021;

CN202110704068.8, application date: June 24, 2021.

technical field

The present invention relates to a class of 5,6,7,8-tetrahydropyridine[3,4-d]pyrimidine compounds, in particular to a compound represented by formula (II) or a pharmaceutically acceptable salt thereof.

Background technique

RAS oncogene mutations are the most common mutations in human cancers, with NRAS, HRAS and KRAS mutations in the RAS family causing nearly a quarter of all human cancers, making it one of the most common genetic mutations associated with cancer . Covers almost all cancer types and causes 1 million deaths worldwide each year. Of these, KRAS is the most common oncogene (85% of all RAS mutations), present in 90% of pancreatic cancers, 30-40% of colon cancers, and 15-20% of lung cancers (mostly non-small cell lung cancers). ). Depending on the specific mutation present, G12C, G12D, and G12R are the most common KRAS mutations. In addition, there are G12A, G12S, G12V and so on.

As one of the most common oncogene mutations in cancer patients, KRAS gene mutation is also a well-known "undruggable" target. The full name of KRAS gene is Kirsten rats arcomaviral oncogene homolog, which is "Kirsten rat sarcoma virus oncogene homolog". The protein encoded by the KARS gene is a small GTPase, which belongs to the RAS superprotein family. KRAS is like a molecular switch, which can control the path of regulating cell growth when it is normal; after KRAS gene mutation, it can independently transmit growth and proliferation signals to downstream pathways independently of upstream growth factor receptor signals, resulting in uncontrolled cell growth and tumor progression. At the same time, whether the KRAS gene has mutation is also an important indicator of tumor prognosis.

Currently, small molecules that directly target KRAS mutations are mainly concentrated in the field of KRAS ^G12C . Among them, Amgen's AMG510 and Mirati Therapeutics' MRTX849 have shown good therapeutic effects on KRAS ^G12C- mutated tumor patients in clinical studies. With the unremitting efforts of scientists and drug R&D personnel, people have also witnessed the milestone of breaking through KRAS's "undruggable" step by step. In the future, more KRAS-targeting therapies will be developed, such as KRAS ^G12D , to save the lives of more cancer patients.

SUMMARY OF THE INVENTION

The present invention provides a compound represented by formula (II) or a pharmaceutically acceptable salt thereof

in,

is selected from single bond or double bond;

T ₁ is selected from CR ₇ R ₈ , NR ₉ and O;

when

is selected from single bond, T is selected from _CH and N;

when

is selected from _double bonds, and T is selected from C;

L ₁ is selected from -CH ₂ - and a bond;

R ₁ , R ₂ , R ₃ , R ₄ and R ₅ are each independently selected from H and C _1-3 alkyl optionally substituted with ₁ , 2 or 3 _Ra ;

R ₆ is selected from C _6-10 aryl and 5-10 membered heteroaryl, the C _6-10 aryl and 5-10 membered heteroaryl are optionally surrounded by 1, 2, 3, 4 or 5 R _b replace;

R ₇ and R ₈ are each independently selected from H, CH ₃ and NH ₂ ;

R ₉ is selected from H and CH ₃ ;

R ₁₀ is selected from H, C _1-3 alkyl, C _1-3 alkoxy and cyclopropyl, the C _1-3 alkyl, C _1-3 alkoxy and cyclopropyl are optionally replaced by 1, 2 or 3 R _c substitutions;

R ₁₁ is selected from 4-8 membered heterocycloalkyl and

The 4-8 membered heterocycloalkyl and

optionally substituted with 1, 2 or 3 _Rd ;

Structural units

selected from 5-6 membered heterocycloalkenyl;

requirement is,

1) R ₁ and R ₂ form a ring with the connected atoms, making the structural unit

form

2) Alternatively, R ₁ and R ₂ form a ring with the connected atoms, making the structural unit

form

3) Alternatively, R ₁ and R ₄ form a ring with the connected atoms, making the structural unit

form

4) Alternatively, R ₄ and R ₅ form a ring with the connected atoms, making the structural unit

form

5) Alternatively, R ₂ and R ₇ form tetrahydropyrrolidinyl with the attached atoms;

6) Alternatively, R ₂ and R ₃ and the connected atom form a C _3-5 membered cycloalkyl;

7) Alternatively, R ₇ and R ₈ form a 4-5 membered heterocycloalkyl with the attached atom;

m is selected from 0, 1 or 2;

n is selected from 0, 1 or 2;

p is selected from 0, 1 or 2;

q is selected from 1, 2 or 3;

r is selected from 1 or 2;

s is selected from 1, 2 or 3;

each R _a is independently selected from F, Cl, Br and I;

Each R _b is independently selected from F, Cl, Br, I, OH, NH ₂ , CN, C _1-3 alkyl, C _1-3 alkoxy, C _2-3 alkynyl, C _2-3 alkene and C _3-5 cycloalkyl, the C _1-3 alkyl, C _1-3 alkoxy, C _2-3 alkynyl, C _2-3 alkenyl and C _3-5 cycloalkyl are optional replaced by 1, 2 or 3 Rs;

each R _c is independently selected from F, Cl, Br and I;

Each R _d is independently selected from H, F, Cl, Br, OH, CN, C _1-3 alkyl, C _1-3 alkoxy and -C _1-3 alkyl-OC(=O)-C _1-3 Alkylamino; each R is independently selected from F, Cl and Br.

In some aspects of the invention, the structural unit

selected from

Other variables are as defined in the present invention.

In some aspects of the invention, the structural unit

selected from

Other variables are as defined in the present invention.

In some aspects of the invention, the structural unit

selected from

Other variables are as defined in the present invention.

In some aspects of the invention, the structural unit

selected from

Other variables are as defined in the present invention.

In some aspects of the invention, the R ₂ and R ₇ form with the attached atoms

Other variables are as defined in the present invention.

In some aspects of the invention, the R ₂ and R ₃ form with the attached atoms

Other variables are as defined in the present invention.

In some aspects of the invention, the _R7 and _R8 form with the attached atom

Other variables are as defined in the present invention.

In some aspects of the invention, the structural unit

selected from

Other variables are as defined in the present invention.

In some aspects of the invention, the structural unit

selected from

Other variables are as defined in the present invention.

In some aspects of the present invention, each R _b is independently selected from F, Cl, Br, I, OH, NH ₂ , CN, CH ₃ , CH ₂ CH ₃ , OCH ₃ , OCH ₂ CH ₃ , - CH=CH ₂ , -CH ₂ -CH=CH ₂ , -C≡CH and cyclopropyl, the CH ₃ , CH ₂ CH ₃ , OCH ₃ , OCH ₂ CH ₃ , -CH=CH ₂ , -CH ₂ -CH= _CH2 , -C≡CH and cyclopropyl are optionally substituted with 1, 2 or 3 R, other variables are as defined in the present invention.

In some aspects of the invention, each R _b is independently selected from F, Cl, OH, NH ₂ , CN, CH ₃ , CF ₃ , CH ₂ CH ₃ and , -C≡CH and cyclopropyl, Other variables are as defined in the present invention.

In some aspects of the present invention, each R _b is independently selected from F, OH, NH ₂ , CH ₃ , CF ₃ , CH ₂ CH ₃ and -C≡CH, and other variables are as defined herein.

In some embodiments of the present invention, the R ₆ is selected from phenyl, pyridyl, naphthyl, indolyl and indazolyl, and said phenyl, pyridyl, naphthyl, indolyl and indazolyl are any Optionally substituted with 1, 2, 3, 4 or 5 R _b , other variables are as defined in the present invention.

In some aspects of the invention, the R ₆ is selected from

Other variables are as defined in the present invention.

In some aspects of the invention, the R ₆ is selected from

Other variables are as defined in the present invention.

In some aspects of the invention, the R ₆ is selected from

Other variables are as defined in the present invention.

In some aspects of the invention, the R ₆ is selected from

Other variables are as defined in the present invention.

In some aspects of the invention, the R ₆ is selected from

Other variables are as defined in the present invention.

In some aspects of the invention, the R ₆ is selected from

Other variables are as defined in the present invention.

In some embodiments of the present invention, said R ₁₀ is selected from H, CH ₃ , OCH ₃ and cyclopropyl, said CH ₃ , OCH ₃ and cyclopropyl are optionally substituted with 1, 2 or 3 R _c , Other variables are as defined in the present invention.

In some aspects of the present invention, the R ₁₀ is selected from H and CH ₃ , and other variables are as defined herein.

In some aspects of the invention, each R _d is independently selected from H, F, Cl, Br, OH, CN, CH ₃ , CH ₂ CH ₃ , CH ₂ CF ₃ , OCH ₃ , OCF ₃ and

Other variables are as defined in the present invention.

In some aspects of the present invention, each R _d is independently selected from H, F, Cl, Br, OH, CN, CH ₃ , CH ₂ CF ₃ , OCH ₃

Other variables are as defined in the present invention.

In some embodiments of the present invention, said R ₁₁ is selected from tetrahydropyrrolyl and hexahydro-1H-pyrrolizinyl, said tetrahydropyrrolyl and hexahydro-1H-pyrrolizinyl being separated by 1, 2 or 3 R _d substitutions, other variables are as defined in the present invention.

In some aspects of the invention, the R ₁₁ is selected from

Other variables are as defined in the present invention.

The present invention provides a compound represented by formula (I) or a pharmaceutically acceptable salt thereof

in,

is selected from single bond or double bond;

T ₁ is selected from CR ₇ R ₈ and NR ₉ ;

when

is selected from single bond, T is selected from _CH and N;

when

is selected from _double bonds, and T is selected from C;

R ₁ , R ₂ , R ₃ , R ₄ and R ₅ are each independently selected from H and C _1-3 alkyl optionally substituted with ₁ , 2 or 3 _Ra ;

R is selected from phenyl and naphthyl optionally substituted with ₁ , ₂ , 3, 4 or 5 R;

R ₇ and R ₈ are each independently selected from H, CH ₃ and NH ₂ ;

R ₉ is selected from H and CH ₃ ;

R ₁₀ is selected from H, C _1-3 alkyl, C _1-3 alkoxy and cyclopropyl, the C _1-3 alkyl, C _1-3 alkoxy and cyclopropyl are optionally replaced by 1, 2 or 3 R _c substitutions;

R ₁₁ is selected from tetrahydropyrrolyl and hexahydro-1H-pyrrolizinyl substituted with 1, 2 or 3 R _d ;

requirement is,

1) R ₁ and R ₂ form a ring with the connected atoms, making the structural unit

form

2) Or, R ₁ and R ₄ are connected with the atoms to form a ring to make the structural unit

form

3) Alternatively, R ₄ and R ₅ form a ring with the connected atoms, making the structural unit

form

4) Alternatively, R ₂ and R ₇ form tetrahydropyrrolidinyl with the attached atom;

5) Alternatively, R ₂ and R ₃ and the connected atoms form a C _3-5 membered cycloalkyl;

6) Alternatively, R ₇ and R ₈ form a 4-5 membered heterocycloalkyl with the attached atom;

m is selected from 0, 1 or 2;

n is selected from 0, 1 or 2;

p is selected from 1 or 2;

q is selected from 1, 2 or 3;

r is selected from 1 or 2;

s is selected from 1, 2 or 3;

each R _a is independently selected from F, Cl, Br and I;

each R _b is independently selected from F, Cl, Br, I, OH, NH ₂ , CN, CH ₃ , CF ₃ and OCH ₃ ;

each R _c is independently selected from F, Cl, Br and I;

Each _Rd is independently selected from H, F, Cl, Br and _CH3 .

In some aspects of the invention, the structural unit

selected from

Other variables are as defined in the present invention.

In some aspects of the invention, the structural unit

selected from

Other variables are as defined in the present invention.

In some aspects of the invention, the structural unit

selected from

Other variables are as defined in the present invention.

In some aspects of the invention, the R ₂ and R ₇ form with the attached atoms

Other variables are as defined in the present invention.

In some aspects of the invention, the R ₂ and R ₃ form with the attached atoms

Other variables are as defined in the present invention.

In some aspects of the invention, the _R7 and _R8 form with the attached atom

Other variables are as defined in the present invention.

In some aspects of the invention, the structural unit

selected from

Other variables are as defined in the present invention.

In some aspects of the invention, the R ₆ is selected from

Other variables are as defined in the present invention.

In some embodiments of the present invention, said R ₁₀ is selected from H, CH ₃ , OCH ₃ and cyclopropyl, said CH ₃ , OCH ₃ and cyclopropyl are optionally substituted with 1, 2 or 3 R _c , Other variables are as defined in the present invention.

In some aspects of the present invention, the R ₁₀ is selected from H and CH ₃ , and other variables are as defined herein.

In some aspects of the invention, the R ₁₁ is selected from

Other variables are as defined in the present invention.

In some embodiments of the invention, the compound, or a pharmaceutically acceptable salt thereof, is selected from

in,

is selected from single bond or double bond;

T ₂ and R ₆ are as defined in the present invention.

In some embodiments of the invention, the compound, or a pharmaceutically acceptable salt thereof, is selected from

in,

T2 is as defined in the present invention _;

R _b1 , R _b2 , R _b3 , R _b4 , R _b5 , R _b6 and R _b7 are each independently selected from H, F, Cl, Br, I, OH, NH ₂ , CN, C _1-3 alkyl, C _1-3 alkoxy, C _2-3 alkynyl, C _2-3 alkenyl and C _3-5 cycloalkyl, the C _1-3 alkyl, C _1-3 alkoxy, C _2-3 Alkynyl, _C2-3alkenyl and _{C3-5cycloalkyl} are optionally substituted with 1, 2 or 3 R;

R is as defined in the present invention.

In some aspects of the invention, the R _b1 , R _b2 , R _b3 , R _b4 , R _b5 , R _b6 and R _b7 are each independently selected from H, F, Cl, Br, I, OH, NH ₂ , CN, _CH3 , _CH2CH3 , _OCH3 , _OCH2CH3 , -CH _{= CH2} , _-CH2 - _CH = _CH2 , _-C≡CH and cyclopropyl, the _CH3 , CH2CH ₃ , OCH ₃ , OCH ₂ CH ₃ , -CH=CH ₂ , -CH ₂ -CH=CH ₂ , -CH=CH ₂ and cyclopropyl are optionally substituted by 1, 2 or 3 R, other variables are as in invention is defined.

In some aspects of the invention, the R _b1 , R _b2 , R _b3 , R _b4 , R _b5 , R _b6 and R _b7 are each independently selected from H, F, Cl, OH, NH ₂ , CN, CH ₃ , _CF3 , _CH2CH3 , _-C≡CH and cyclopropyl, other variables are as defined in the present invention.

There are also some solutions of the present invention that are formed by any combination of the above variables.

The present invention also provides a compound represented by the following formula or a pharmaceutically acceptable salt thereof, wherein the compound is selected from the group consisting of

In some aspects of the invention, the compound or a pharmaceutically acceptable salt thereof, the compound is selected from,

In some embodiments of the invention, the compound, or a pharmaceutically acceptable salt thereof, is selected from

The present invention also provides following synthetic method:

method 1:

in,

R ₆ and R ₁₁ are as defined in the present invention.

Method 2:

The present invention also provides the following test methods:

Method 2. Anti-proliferative effects of compounds in tumor cell lines AsPC-1 and GP2D

Research purposes

In this experiment, the inhibitory effect of the compound on cell proliferation was studied by detecting the effect of the compound on the cell viability in vitro in the tumor cell lines AsPC-1 and GP2D.

Experimental Materials

Table 1. Experimental Materials

cell line	tumor type	Growth characteristics	Cultivation method
AsPC-1	Pancreatic cancer	adherent growth	RPMI 1640+10% FBS
GP2D	colon cancer	adherent growth	DMEM+10%FBS+2mM L-glutamine

Ultra Low Cluster-96 well plate (Corning-7007)

Greiner CELLSTAR 96-well plate (#655090)

Promega CellTiter-Glo 3D Luminescence Cell Viability Assay Kit (Promega-G9683)

2104-10 EnVision Plate Reader, PerkinElmer

RPMI 1640, DMEM, PBS (phosphate buffered saline), FBS (fetal bovine serum), Antibiotic-antimycotic (antibiotic-antifungal), L-glutamine (L-glutamine), DMSO (dimethyl sulfoxide)

Experimental methods and steps

cell culture

The tumor cell lines were cultured in a 37°C, 5% CO2 incubator according to the culture conditions indicated in the culture method. Periodically passaged, cells in logarithmic growth phase were taken for plating.

Cell plating

Cells were stained with trypan blue and viable cells were counted.

Adjust the cell concentration to an appropriate concentration.

Table 2. Cell Concentrations

cell line	Density (per hole)
AsPC-1	7000 cells
GP2D	8000 cells

135 μL of cell suspension was added to each well of the ULA culture plate, and the same volume of culture medium without cells was added to the blank control air.

Immediately after plating, the ULA plates were centrifuged at 1000 rpm for 10 minutes at room temperature. NOTE: After centrifugation, be careful not to cause unnecessary shaking.

The plates were incubated overnight in an incubator at 37°C, 5% _CO2 , and 100% relative humidity.

Preparation of 10X Compound Working Solution and Compound Treatment of Cells (Day 1)

After preparing the 10X compound working solution (DMSO 10X working solution), add 15 μL of 10X compound working solution to the ULA culture plate respectively, and add 15 μL of DMSO-cell culture solution mixture to the vehicle control and blank control.

Return the 96-well cell plate to the incubator for 120 hours.

Cell spheroidization was observed every day until the end of the experiment.

CellTiter-Glo Luminescence Assay for Cell Viability (Day 5)

The following steps were performed according to the instructions of the Promega CellTiter-Glo 3D Luminescence Cell Viability Assay Kit (Promega#G9683).

Add 150 μL (equivalent to the volume of cell culture medium in each well) of CellTiter-Glo 3D reagent to each well. Wrap the cell plate in aluminum foil to protect from light.

Shake the plate on an orbital shaker for 5 minutes.

Mix the air mixture carefully by pipetting up and down 10 times. Make sure the spheroids are sufficiently detached before proceeding to the next step.

The solution in the ULA plate was then transferred to a black bottom plate (#655090) and left at room temperature for 25 minutes to stabilize the luminescent signal. Luminescent signals were detected on a 2104 EnVision plate reader.

data analysis

The inhibition rate (IR) of the tested compound was calculated by the following formula: IR(%)=(1−(RLU compound−RLU blank control)/(RLU vehicle control−RLU blank control))*100%. The inhibition rates of different concentrations of compounds were calculated in Excel, and then the GraphPad Prism software was used to plot the inhibition curves and calculate the relevant parameters, including the minimum inhibition rate, the maximum inhibition rate and IC ₅₀ .

Method 3. In vivo pharmacokinetic experiments

Purpose:

The purpose of this experiment is to investigate the pharmacokinetic characteristics of the compounds of the present invention under oral and intravenous injection in SD mice.

experimental method:

The test compound was mixed with 10% dimethyl sulfoxide/60% polyethylene glycol 400/30% aqueous solution, vortexed and sonicated to prepare a clear solution of about 1 mg/mL, which was filtered through a microporous membrane for use. Male SD mice aged 7 to 10 weeks were selected, and the candidate compound solution was administered intravenously at a dose of about 3 mg/kg. Candidate compound solutions are administered orally at a dose of approximately 30 mg/kg. Whole blood was collected for a certain period of time, plasma was prepared, drug concentration was analyzed by LC-MS/MS method, and pharmacokinetic parameters were calculated by Phoenix WinNonlin software (Pharsight, USA).

Method 4. In vivo pharmacodynamic experiments

Purpose:

In vivo pharmacodynamics study of human colorectal cancer GP2D cells in nude mice subcutaneously transplanted in Balb/c Nude mouse model

experimental method:

Cell culture: Human colorectal cancer GP2D cells were cultured in monolayer in vitro, and the culture conditions were DMEM/F12 medium with 20% fetal bovine serum, 1% double antibody, and a 37°C 5% carbon dioxide incubator. Conventional digestion with trypsin-EDTA was performed twice a week for passage. When the cell saturation is 80%-90% and the number reaches the requirement, collect the cells, count them, resuspend in an appropriate amount of PBS, and add Matrigel 1:1 to obtain a cell suspension with a cell density of 25x 10 ⁶ cells/mL.

Cell seeding: 0.2 mL (5×10 ⁶ cells/mouse) Mia PaCa-2 cells (plus Matrigel, 1:1 by volume) were subcutaneously seeded on the right back of each mouse.

Experimental operation: When the average tumor volume reached ^190mm3 , random grouping was conducted according to the tumor volume, 6 rats in each group, the blank group was given a dose of 0, and the test group was given a dose gradient, orally, for 22 days, twice a day. .

Tumor measurements and experimental indicators:

Tumor diameters were measured with vernier calipers twice a week. The calculation formula of tumor volume is: V=0.5a×b ² , a and b represent the long and short diameters of the tumor, respectively.

The antitumor efficacy of the compounds was evaluated by TGI (%) or relative tumor proliferation rate T/C (%). Relative tumor proliferation rate T/C (%)=TRTV/CRTV×100% (TRTV: RTV of treatment group; CRTV: RTV of negative control group). The relative tumor volume (RTV) is calculated according to the results of tumor measurement, and the calculation formula is RTV=V _t /V ₀ , where V ₀ is the average tumor volume measured during group administration (ie, D ₀ ), V _t The mean tumor volume at one measurement, TRTV and CRTV take the same day data.

TGI (%), reflecting tumor growth inhibition rate. TGI(%)=[(1-(average tumor volume at the end of administration of a certain treatment group-average tumor volume at the beginning of administration of this treatment group))/(average tumor volume at the end of treatment in the solvent control group-the start of treatment in the solvent control group time average tumor volume)] × 100%.

technical effect

The compound of the present invention has good binding effect and inhibitory effect on KRAS ^G12D protein, has good cell proliferation inhibitory activity on KRAS ^G12D mutant cells, and has excellent tumor inhibitory effect. In addition, the compounds of the present invention have better pharmacokinetic characteristics.

Related Definitions

Unless otherwise specified, the following terms and phrases used herein are intended to have the following meanings. A particular term or phrase should not be considered indeterminate or unclear without specific definitions, but should be understood in its ordinary meaning. When a trade name appears herein, it is intended to refer to its corresponding commercial product or its active ingredient.

As used herein, the term "pharmaceutically acceptable" refers to those compounds, materials, compositions and/or dosage forms that, within the scope of sound medical judgment, are suitable for use in contact with human and animal tissue , without excessive toxicity, irritation, allergic reactions or other problems or complications, commensurate with a reasonable benefit/risk ratio.

The term "pharmaceutically acceptable salts" refers to salts of the compounds of the present invention, prepared from compounds with specific substituents discovered by the present invention and relatively non-toxic acids or bases. When compounds of the present invention contain relatively acidic functional groups, base addition salts can be obtained by contacting such compounds with a sufficient amount of base in neat solution or in a suitable inert solvent. Pharmaceutically acceptable base addition salts include sodium, potassium, calcium, ammonium, organic amine or magnesium salts or similar salts. When compounds of the present invention contain relatively basic functional groups, acid addition salts can be obtained by contacting such compounds with a sufficient amount of acid in neat solution or in a suitable inert solvent. Examples of pharmaceutically acceptable acid addition salts include inorganic acid salts including, for example, hydrochloric acid, hydrobromic acid, nitric acid, carbonic acid, bicarbonate, phosphoric acid, monohydrogen phosphate, dihydrogen phosphate, sulfuric acid, Hydrogen sulfate, hydroiodic acid, phosphorous acid, etc.; and organic acid salts including, for example, acetic acid, propionic acid, isobutyric acid, maleic acid, malonic acid, benzoic acid, succinic acid, suberic acid, Similar acids such as fumaric, lactic, mandelic, phthalic, benzenesulfonic, p-toluenesulfonic, citric, tartaric, and methanesulfonic acids; also include salts of amino acids such as arginine, etc. , and salts of organic acids such as glucuronic acid. Certain specific compounds of the present invention contain both basic and acidic functional groups and thus can be converted into either base or acid addition salts.

The pharmaceutically acceptable salts of the present invention can be synthesized from the acid or base containing parent compound by conventional chemical methods. Generally, such salts are prepared by reacting the free acid or base form of these compounds with a stoichiometric amount of the appropriate base or acid in water or an organic solvent or a mixture of the two.

The compounds of the present invention may exist in specific geometric or stereoisomeric forms. The present invention contemplates all such compounds, including cis and trans isomers, (-)- and (+)-enantiomers, (R)- and (S)-enantiomers, diastereomers isomers, (D)-isomers, (L)-isomers, and racemic mixtures thereof and other mixtures, such as enantiomerically or diastereomerically enriched mixtures, all of which belong to this within the scope of the invention. Additional asymmetric carbon atoms may be present in substituents such as alkyl groups. All such isomers, as well as mixtures thereof, are included within the scope of the present invention.

The compounds of the present invention may contain unnatural proportions of atomic isotopes at one or more of the atoms that constitute the compound. For example, compounds can be labeled with radioisotopes, such as tritium ( ³ H), iodine-125 ( ¹²⁵ I) or C-14 ( ¹⁴ C). For another example, deuterated drugs can be formed by replacing hydrogen with deuterium, and the bond formed by deuterium and carbon is stronger than the bond formed by ordinary hydrogen and carbon. Compared with undeuterated drugs, deuterated drugs can reduce toxic side effects and increase drug stability. , enhance the efficacy, prolong the biological half-life of drugs and other advantages. All transformations of the isotopic composition of the compounds of the present invention, whether radioactive or not, are included within the scope of the present invention.

The terms "optional" or "optionally" mean that the subsequently described event or circumstance may, but need not, occur, and that the description includes instances where said event or circumstance occurs and instances in which it does not. .

The term "substituted" means that any one or more hydrogen atoms on a specified atom are replaced by a substituent, which may include deuterium and hydrogen variants, as long as the valence of the specified atom is normal and the substituted compound is stable. When the substituent is oxygen (ie =O), it means that two hydrogen atoms are substituted. Oxygen substitution does not occur on aromatic groups. The term "optionally substituted" means that it may or may not be substituted, and unless otherwise specified, the type and number of substituents may be arbitrary on a chemically achievable basis.

When any variable (eg, R) occurs more than once in the composition or structure of a compound, its definition in each case is independent. Thus, for example, if a group is substituted with 0-2 Rs, the group may optionally be substituted with up to two Rs, with independent 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 a linking group is 0, such as -(CRR) ₀ -, it means that the linking group is a single bond.

When one of the variables is selected from a single bond, it means that the two groups connected thereto are directly connected, for example, when L in A-L-Z represents a single bond, it means that the structure is actually A-Z.

When the listed linking group does not indicate its direction of attachment, the direction of attachment is arbitrary, for example,

The linking group L in the middle is -MW-, at this time -MW- can connect ring A and ring B in the same direction as the reading order from left to right.

It is also possible to connect ring A and ring B in the opposite direction to the reading order from left to right.

Combinations of the linking groups, substituents and/or variants thereof are permissible only if such combinations result in stable compounds.

Unless otherwise specified, when a group has one or more attachable sites, any one or more sites in the group can be linked to other groups by chemical bonds. When the connection method of the chemical bond is not positioned and there is an H atom at the connectable site, when the chemical bond is connected, the number of H atoms at the site will decrease correspondingly with the number of chemical bonds connected to the corresponding valence. the group. The chemical bond connecting the site to other groups can be represented by straight solid line bonds

straight dotted key

or wavy lines

express. For example, a straight solid bond in -OCH ₃ indicates that it is connected to other groups through the oxygen atom in this group;

The straight dashed bond in the group indicates that it is connected to other groups through the two ends of the nitrogen atom in the group;

The wavy line in the phenyl group indicates that it is connected to other groups through the 1 and 2 carbon atoms in the phenyl group;

Indicates that any linkable site on the piperidinyl group can be connected to other groups through a chemical bond, including at least

These 4 connection methods, even if the H atom is drawn on -N-, but

still includes

The group in this connection method is only that when one chemical bond is connected, the H at the site will be correspondingly reduced by one to become the corresponding monovalent piperidinyl group.

Use solid wedge keys unless otherwise specified

and wedge-dotted keys

Indicate the absolute configuration of a stereocenter, using a straight solid key

and straight dashed keys

Indicate the relative configuration of the stereocenter, with a wavy line

Represents a solid wedge key

or wedge-dotted key

or with wavy lines

Represents a straight solid key

or straight dashed key

Unless otherwise specified, the term "C _1-3 alkyl" is used to denote a straight or branched chain saturated hydrocarbon group consisting of 1 to 3 carbon atoms. The C _1-3 alkyl group includes C _1-2 and C _2-3 alkyl groups, etc.; it can be monovalent (eg methyl), divalent (eg methylene) or multivalent (eg methine) . Examples of _C1-3 alkyl groups include, but are not limited to, methyl (Me), ethyl (Et), propyl (including n-propyl and isopropyl), and the like.

Unless otherwise specified, the term " _C1-3alkoxy " refers to those alkyl groups containing 1 to 3 carbon atoms attached to the remainder of the molecule through an oxygen atom. The C _1-3 alkoxy group includes C _1-2 , C _2-3 , C ₃ and C ₂ alkoxy and the like. Examples of C _1-3 alkoxy groups include, but are not limited to, methoxy, ethoxy, propoxy (including n-propoxy and isopropoxy), and the like.

Unless otherwise specified, the term "C _1-3 alkylamino" refers to those alkyl groups containing 1 to 3 carbon atoms attached to the remainder of the molecule through an amino group. The C _1-3 alkylamino groups include C _1-2 , C ₃ and C ₂ alkylamino groups and the like. Examples of C _1-3 alkylamino include, but are not limited to, -NHCH ₃ , -N(CH ₃ ) ₂ , -NHCH ₂ CH ₃ , -N(CH ₃ )CH ₂ CH ₃ , -NHCH ₂ CH ₂ CH ₃ , - NHCH ₂ (CH ₃ ) ₂ and the like.

Unless otherwise specified, "C _2-3 alkenyl" is used to denote a straight or branched chain hydrocarbon group consisting of 2 to 3 carbon atoms containing at least one carbon-carbon double bond, a carbon-carbon double bond can be located anywhere in the group. The C _2-3 alkenyl group includes C ₃ and C ₂ alkenyl groups; the C _2-3 alkenyl group may be monovalent, divalent or multivalent. Examples of C _2-3 alkenyl groups include, but are not limited to, vinyl, propenyl, and the like.

Unless otherwise specified, "C _2-3 alkynyl" is used to denote a straight or branched chain hydrocarbon group consisting of 2 to 3 carbon atoms containing at least one carbon-carbon triple bond, a carbon-carbon triple bond can be located anywhere in the group. It can be monovalent, bivalent or multivalent. The C _2-3 alkynyl groups include C ₃ and C ₂ alkynyl groups. Examples of _C2-3alkynyl groups include, but are not limited to, ethynyl, propynyl, and the like.

Unless otherwise specified, the term "4-5 membered heterocycloalkyl" by itself or in combination with other terms denotes a saturated monocyclic group consisting of 4 to 5 ring atoms, 1, 2, 3 or 4 ring atoms, respectively are heteroatoms independently selected from O, S, and N, and the remainder are carbon atoms, where the nitrogen atom is optionally quaternized, and the nitrogen and sulfur heteroatoms are optionally oxidized (ie, NO and S(O) _p , p is 1 or 2). Furthermore, with respect to the "4-5 membered heterocycloalkyl", a heteroatom may occupy the position of attachment of the heterocycloalkyl to the remainder of the molecule. The 4-5 membered heterocycloalkyl includes 4 and 5 membered heterocycloalkyl. Examples of 4-5 membered heterocycloalkyl include, but are not limited to, azetidinyl, oxetanyl, thietanyl, pyrrolidinyl, pyrazolidinyl, imidazolidinyl, tetrahydrothienyl ( Including tetrahydrothiophen-2-yl and tetrahydrothiophen-3-yl, etc.) or tetrahydrofuranyl (including tetrahydrofuran-2-yl, etc.) and the like.

Unless otherwise specified, "C _3-5 cycloalkyl" means a saturated cyclic hydrocarbon group consisting of 3 to 5 carbon atoms, which is a monocyclic ring system, said C _3-5 cycloalkyl including C _{3 -4} and C _4-5 cycloalkyl, etc.; it may be monovalent, divalent or polyvalent. Examples of _C3-5 cycloalkyl include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, and the like.

Unless otherwise specified, the terms "C _6-10 aryl ring" and "C _6-10 aryl group" can be used interchangeably in the present invention, and the term "C _6-10 aryl ring" or "C _6-10 aryl group" means by A cyclic hydrocarbon group composed of 6 to 10 carbon atoms with a conjugated π-electron system, which may be a monocyclic, fused bicyclic or fused tricyclic system, wherein each ring is aromatic. It may be monovalent, divalent or polyvalent, and _C6-10 aryl groups include _C6-9 , _C9 , _C10 and _C6 aryl groups and the like. Examples of _C6-10 aryl groups include, but are not limited to, phenyl, naphthyl (including 1-naphthyl and 2-naphthyl, and the like).

Unless otherwise specified, the terms "5-10-membered heteroaryl ring" and "5-10-membered heteroaryl" can be used interchangeably in the present invention, and the term "5-10-membered heteroaryl" refers to a ring consisting of 5 to 10 rings. A cyclic group composed of atoms with a conjugated π-electron system, wherein 1, 2, 3 or 4 ring atoms are heteroatoms independently selected from O, S and N, and the rest are carbon atoms. It can be a monocyclic, fused bicyclic or fused tricyclic ring system, wherein each ring is aromatic. Where the nitrogen atom is optionally quaternized, the nitrogen and sulfur heteroatoms may be optionally oxidized (ie, NO and S(O) _p , p is 1 or 2). A 5-10 membered heteroaryl group can be attached to the rest of the molecule through a heteroatom or a carbon atom. The 5-10-membered heteroaryl groups include 10-membered, 9-membered, 9-10-membered, 5-8-membered, 5-7-membered, 5-6-membered, 5-membered, and 6-membered heteroaryl groups, and the like. Examples of the 5-10 membered heteroaryl group include, but are not limited to, pyrrolyl (including N-pyrrolyl, 2-pyrrolyl and 3-pyrrolyl, etc.), pyrazolyl (including 2-pyrazolyl and 3-pyrrolyl, etc.) azolyl, etc.), imidazolyl (including N-imidazolyl, 2-imidazolyl, 4-imidazolyl and 5-imidazolyl, etc.), oxazolyl (including 2-oxazolyl, 4-oxazolyl and 5- oxazolyl, etc.), triazolyl (1H-1,2,3-triazolyl, 2H-1,2,3-triazolyl, 1H-1,2,4-triazolyl and 4H-1, 2,4-triazolyl, etc.), tetrazolyl, isoxazolyl (3-isoxazolyl, 4-isoxazolyl and 5-isoxazolyl, etc.), thiazolyl (including 2-thiazolyl , 4-thiazolyl and 5-thiazolyl, etc.), furyl (including 2-furyl and 3-furyl, etc.), thienyl (including 2-thienyl and 3-thienyl, etc.), pyridyl (including 2- -pyridyl, 3-pyridyl and 4-pyridyl, etc.), pyrazinyl, pyrimidinyl (including 2-pyrimidinyl and 4-pyrimidinyl, etc.), benzothiazolyl (including 5-benzothiazolyl, etc.) , purinyl, benzimidazolyl (including 2-benzimidazolyl, etc.), benzoxazolyl, indolyl (including 5-indolyl, etc.), isoquinolinyl (including 1-isoquinolinyl, etc.) and 5-isoquinolinyl, etc.), quinoxalinyl (including 2-quinoxalinyl and 5-quinoxalinyl, etc.) or quinolinyl (including 3-quinolinyl and 6-quinolinyl, etc.) .

Unless otherwise specified, the term "4-8 membered heterocycloalkyl" by itself or in combination with other terms denotes a saturated cyclic group consisting of 4 to 8 ring atoms, respectively, of which 1, 2, 3 or 4 ring atoms are heteroatoms independently selected from O, S, and N, and the remainder are carbon atoms, where the nitrogen atom is optionally quaternized, and the nitrogen and sulfur heteroatoms are optionally oxidized (ie, NO and S(O) _p , p is 1 or 2). It includes monocyclic and bicyclic ring systems, wherein bicyclic ring systems include spiro, paracyclic and bridged rings. Additionally, with respect to the "4-8 membered heterocycloalkyl", a heteroatom may occupy the position of attachment of the heterocycloalkyl to the remainder of the molecule. The 4-8 membered heterocycloalkyl includes 4-6 membered, 5-6 membered, 4 membered, 5 membered and 6 membered heterocycloalkyl and the like. Examples of 4-8 membered heterocycloalkyl include, but are not limited to, azetidinyl, oxetanyl, thietanyl, pyrrolidinyl, pyrazolidinyl, imidazolidinyl, tetrahydrothienyl ( Including tetrahydrothiophen-2-yl and tetrahydrothiophen-3-yl, etc.), tetrahydrofuranyl (including tetrahydrofuran-2-yl, etc.), tetrahydropyranyl, piperidinyl (including 1-piperidinyl, 2- piperidinyl and 3-piperidyl, etc.), piperazinyl (including 1-piperazinyl and 2-piperazinyl, etc.), morpholinyl (including 3-morpholinyl and 4-morpholinyl, etc.), Dioxanyl, dithianyl, isoxazolidinyl, isothiazolidinyl, 1,2-oxazinyl, 1,2-thiazinyl, hexahydropyridazinyl, homopiperazinyl, homopiperyl pyridyl or dioxepanyl, etc.

Unless otherwise specified, the term "5-6 membered heterocycloalkenyl" by itself or in combination with other terms respectively denotes a partially unsaturated cyclic group consisting of 5 to 6 ring atoms containing at least one carbon-carbon double bond , whose 1, 2, 3 or 4 ring atoms are heteroatoms independently selected from O, S and N, and the rest are carbon atoms, wherein the nitrogen atom is optionally quaternized, and the nitrogen and sulfur heteroatoms may optionally be Oxidation (ie NO and S(O) _p , p is 1 or 2). It includes monocyclic and bicyclic ring systems, wherein bicyclic ring systems include spiro, paracyclic and bridged rings, any ring of this system is non-aromatic. Furthermore, in the case of the "5-6 membered heterocycloalkenyl", a heteroatom may occupy the position of attachment of the heterocycloalkenyl to the rest of the molecule. The 5-6 membered heterocyclenyl includes 5-membered and 6-membered heterocyclenyl and the like. Examples of 5-6 membered heterocycloalkenyl include but are not limited to

Unless otherwise specified, _Cn-n+m or _Cn - _Cn+m includes any particular instance of n to n+ _m carbons, eg _C1-12 includes _C1 , _C2 , C3, C ₄ , C ₅ , C ₆ , C ₇ , C ₈ , C ₉ , C ₁₀ , C ₁₁ , and C ₁₂ , also including any range from n to n+ _m , eg C _1-12 includes C _1-3 , C _1-6 , C _1-9 , C _3-6 , C _3-9 , C _3-12 , C _6-9 , C _6-12 , and C _9-12 , etc.; in the same way, n yuan to n +m-membered means that the number of atoms in the ring is from n to n+m, for example, 3-12-membered ring includes 3-membered ring, 4-membered ring, 5-membered ring, 6-membered ring, 7-membered ring, 8-membered ring, 9-membered ring , 10-membered ring, 11-membered ring, and 12-membered ring, also including any one range from n to n+m, for example, 3-12-membered ring includes 3-6 membered ring, 3-9 membered ring, 5-6 membered ring ring, 5-7 membered ring, 6-7 membered ring, 6-8 membered ring, and 6-10 membered ring, etc.

The compounds of the present invention can be prepared by a variety of synthetic methods well known to those skilled in the art, including the specific embodiments enumerated below, embodiments formed in combination with other chemical synthesis methods, and those well known to those skilled in the art Equivalent to alternatives, preferred embodiments include, but are not limited to, the embodiments of the present invention.

The structure of the compound of the present invention can be confirmed by conventional methods well known to those skilled in the art. If the present invention relates to the absolute configuration of the compound, the absolute configuration can be confirmed by conventional technical means in the art. For example, single crystal X-ray diffraction method (SXRD), the cultured single crystal is collected by Bruker D8 venture diffractometer, the light source is CuKα radiation, and the scanning mode is:

After scanning and collecting relevant data, the crystal structure was further analyzed by the direct method (Shelxs97), and the absolute configuration could be confirmed.

The solvent used in the present invention is commercially available.

Compounds are named according to conventional nomenclature in the art or are used

Software naming, commercially available compounds use supplier catalog names.

Description of drawings

Figure 1. Binding pattern of compound A and KRAS ^G12D protein;

Figure 2. Binding pattern of compound B and KRAS ^G12D protein;

Figure 3. Binding pattern of Compound C and KRAS ^G12D protein.

Detailed ways

The present invention will be described in detail by the following examples, but it does not mean any unfavorable limitation of the present invention. The present invention has been described in detail herein, and specific embodiments thereof have also been disclosed. For those skilled in the art, various changes and modifications can be made to the specific embodiments of the present invention without departing from the spirit and scope of the invention. will be obvious.

Calculation example 1

The molecular docking process was performed by using Maestro (

Glide SP[1] and default options in version 2017-2). The crystal structure PDB:6UT0 of KRAS_G12C in the PDB database was selected, Cys12 was simulated and mutated to Asp12, and after energy optimization, it was used as the docking template. To prepare the protein, hydrogen atoms were added using the Protein Preparation Wizard module of Maestro [2] and the OPLS3 force field was used. For ligand preparation, the 3D structure of the molecule was generated using LigPrep and energy minimization was performed [3], and the small molecule conformation was sampled using the confgen module. Using the ligand of 6UT0 as the centroid, the side length is

The cube docking grid. Place reference compounds during molecular docking. The interaction types between protein receptors and ligands are analyzed, and the interaction types between protein receptors and ligands are analyzed, and then a reasonable docking conformation is selected and saved according to the calculated docking scrore and binding mode.

[1]Glide,

LLC, New York, NY, 2017.

[2] Maestro,

LLC, New York, NY, 2017.

[3]LigPrep,

LLC, New York, NY, 2017.

Conclusion: The compound of the present invention has good binding with KRAS G12D.

Example 1

synthetic route:

Step 1: Synthesis of Compounds 1-2

Under nitrogen protection, 1-1 (72g, 405.34mmol, 1eq), DMAP (49.52g, 405.34mmol, 1eq) and 1-1a (395.31g, 2.03mol, 5eq) were dissolved in TEA (261.72g, 2.59mol, 6.38 eq), the reaction solution was stirred at 80 °C for 12 h. EtOAc (800 mL*2) and H ₂ O (800 mL) were added to the reaction solution for extraction and separation, the organic phase was washed with saturated brine (500 mL), the organic phase was dried, filtered and concentrated to obtain crude product 1-2. MS m/z: 292.1[M+1] ⁺ .

Step 2: Synthesis of Compounds 1-3

Under nitrogen protection, 1-2a (390.64g, 2.34mol, 13.65eq), KI (71.12g, 428.42mmol, 2.5eq) and 1-2 (50g, 171.37mmol, 1eq) _were dissolved in K2CO3 ( _59.21g ) , 428.42mmol, 2.5eq), the reaction solution was stirred at 80°C for 12h. H ₂ O (600 mL) and EtOAc (600 mL*2) were added to the reaction solution for extraction and separation, the organic phase was washed with saturated brine (500 mL), and the organic phase was dried, filtered and concentrated to obtain compound 1-3. MS m/z: 378.1[M+1] ⁺ .

Step 3: Synthesis of Compounds 1-4

Under nitrogen protection, compound 1-3 (10 g, 26.46 mmol, 1 eq) was dissolved in THF (200 mL) at -65 °C, and then LDA (2M, 27.79 mL, 2.1 eq) was slowly added dropwise, and the reaction was performed at -65 °C for 1 hour. A saturated ammonium chloride solution (100 mL) was added to the reaction system, the aqueous phase was extracted with methyl tertiary ether (100 mL*2), and the layers were separated. The organic phases were combined, washed with saturated brine (100 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain the crude product. The crude product was separated by a column machine (mobile phase: ethyl acetate/petroleum ether=0-0.5%), and purified to obtain compound 1-4. ¹ H NMR (400 MHz, CDCl ₃ ) δ=12.08(s, 1H), 7.74-7.73(m, 1H), 7.63-7.58(m, 1H), 7.55-7.53(m, 1H), 7.43(t, J =7.6Hz,1H),7.35-7.33(m,1H),7.22(dd,J=7.6,1.2Hz,1H),4.28(q,J=7.2Hz,2H),3.92-3.88(m,1H) , 3.53-3.48(m,1H),3.46-3.40(m,1H),3.10-3.03(m,1H),2.75-2.68(m,1H),2.38-2.32(m,1H),1.35(t, J=6.8Hz,3H).MS m/z:332.1[M+1] ⁺ .

Step 4: Synthesis of Compounds 1-5

Under nitrogen protection, compound 1-4 (5.9 g, 11.56 mmol, 1 eq) was dissolved in MeOH (59 mL), and compound 1-4a (3.26 g, 17.34 mmol, 1.5 eq) and CH ₃ ONa (2.87 were added in three batches at 0°C) g, 53.17 mmol, 4.6 eq), during which the temperature was maintained at 0-15 °C, and then reacted at 20 °C for 21 hours. 2N hydrochloric acid solution was used to adjust the pH of the system to 5, a solid was precipitated, filtered, the filter cake was washed with methanol (500 mL) and methyl tertiary ether (500 mL) successively, and the filter cake was dried under reduced pressure to obtain the crude product of compound 1-5. The reaction system was directly used in the next step without purification. MS m/z: 358.1[M+1] ⁺ .

Step 5: Synthesis of Compounds 1-6

Under nitrogen protection, compound 1-5 (0.98g, 2.74mmol, 1eq) was dissolved in dichloromethane (10mL), N,N-diisopropylethylamine (1.06g, 8.22mmol, 1.43mL, 3eq) was added, Trifluoromethanesulfonic anhydride (1.55g, 5.48mmol, 903.68μL, 2eq) was slowly added dropwise at 0°C, maintaining the temperature at 0-8°C during the period, and after the dropwise addition was completed, the temperature was slowly raised to 20°C for reaction for 1 hour. Water (50 mL) was added to the reaction system, and the layers were separated. The aqueous phase was extracted with dichloromethane (50 mL*2), and the layers were separated. The organic phases were combined, washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain compound 1-6. This compound was not purified and used directly in the next reaction. MS m/z: 489.9[M+1] ⁺ .

Step 6: Synthesis of Compounds 1-7

Under nitrogen protection, compound 1-6 (3g, 1.84mmol, 1eq) and compound 1-6a (397.78mg, 1.87mmol, 1.02eq) were dissolved in acetonitrile (18mL), and potassium carbonate (507.79mg, 3.67mmol, 2eq) was added. , 10 ℃ reaction for 13 hours. Water (50 mL) was added to the reaction system, the aqueous phase was extracted with methyl tertiary ether (60 mL*2), and the layers were separated. The organic phases were combined, washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain the crude product. The crude product was separated by column chromatography (mobile phase: petroleum ether: ethyl acetate=30:1-10:1), and purified to obtain compound 1-7. ¹ H NMR (400 MHz, CDCl ₃ ) δ 7.74 (d, J=8.1 Hz, 1H), 7.59 (d, J=8.1 Hz, 1H), 7.51 (d, J=7.4 Hz, 1H), 7.43 (t , J＝7.8Hz, 1H), 7.32(t, J＝7.8Hz, 1H), 7.21(d, J＝7.5Hz, 1H), 4.42(d, J＝17.6Hz, 1H), 4.37–4.20(m ,2H),4.08(d,J＝12.6Hz,1H),3.85(d,J＝17.8Hz,1H),3.69–3.59(m,1H),3.57–3.48(m,1H),3.47–3.30( m, 1H), 3.20–3.02 (m, 3H), 2.51 (s, 3H), 2.49 (s, 1H), 2.01–1.87 (m, 3H), 1.81–1.70 (m, 1H), 1.49 (s, 9H). MS m/z: 552.1[M+1] ⁺ .

Step 7: Synthesis of Compounds 1-8

Under nitrogen protection, compound 1-7 (0.74g, 1.34mmol, 1eq) was dissolved in dichloromethane (25mL), m-chloroperoxybenzoic acid (326.53mg, 1.61mmol, 1.2eq) was added at 0°C, and then the temperature was slowly raised to The reaction was carried out at 20°C for 2 hours. Saturated sodium thiosulfate solution (10 mL) was added to the reaction system, and the solution was separated. The aqueous phase was extracted with dichloromethane (20 mL*2), and the layers were separated. The organic phases were combined, washed with saturated sodium bicarbonate solution (10 mL), and separated. The aqueous phase was extracted with dichloromethane (10 mL*2), and the layers were separated. The organic phases were combined, washed with saturated brine (10 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain compound 1-8. This compound was not purified and used directly in the next reaction. MS m/z: 568.2[M+1] ⁺ .

Step 8: Synthesis of Compounds 1-9

Under nitrogen protection, compound 1-8 (0.15g, 264.02μmol, 1eq) and compound 1-8a (50.44mg, 316.83μmol, 1.2eq) were dissolved in toluene (10mL), and sodium tert-butoxide (38.06mg, 0 ℃) was added. 396.04μmol, 1.5eq), and then react at 0°C for 1 hour. Water (5 mL) was added to the reaction system, the aqueous phase was extracted with ethyl acetate (10 mL*3), and the layers were separated. The organic phases were combined, washed with saturated brine (10 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain compound 1-9. This compound was not purified and used directly in the next reaction. MS m/z: 663.2[M+1] ⁺ .

Step 9: Synthesis of Compound 1

Under nitrogen protection, compound 1-9 (0.32 g, 482.49 μmol) was dissolved in methanol (5 mL), hydrochloric acid/methanol (4 M, 15.71 mL, ) was added, and the reaction was carried out at 20° C. for 2 hours. The reaction system was directly concentrated under reduced pressure to obtain the crude product. The crude product was separated by high performance liquid chromatography (column: Waters Xbridge Prep OBD C18 150*40mm*10μm; mobile phase: A (acetonitrile) and B (water, containing 0.05% ammonia and 10 mM ammonium bicarbonate); gradient: B% : 35%-65%, 8 min), purified to give compound 1. ¹ H NMR (400 MHz, CDCl ₃ ) δ=7.74 (d, J=8.0 Hz, 1H), 7.59 (d, J=8.0 Hz, 1H), 7.51 (d, J=7.2 Hz, 1H), 7.43 (t ,J=7.6Hz,1H),7.32(t,J=7.6Hz,1H),7.20(d,J=7.6Hz,1H),5.32-5.19(m,1H),4.40(d,J=16.8Hz ,1H),4.15-4.07(m,2H),4.00-3.92(m,1H),3.86(dd,J=17.6,5.6Hz,1H),3.69(d,J=13.2Hz,1H),3.58( s, 2H), 3.53-3.50(m, 1H), 3.32(d, J=12.0Hz, 1H), 3.27-3.22(m, 2H), 3.16-3.03(m, 4H), 2.99-2.93(m, 1H), 2.50 (d, J=13.2Hz, 1H), 2.32-2.12 (m, 3H), 2.01-1.87 (m, 4H), 1.80 (s, 4H). MS m/z: 563.3[M+1] ⁺ .

Example 2

Step 1: Synthesis of Compound 2-2

Intermediate 2-1 (5.53 g, 16.35 mmol) and Intermediate 1-6a (3.47 g, 16.35 mmol) were dissolved in tetrahydrofuran (50 mL), followed by the addition of N,N-diisopropylethylamine (2.32 g, 17.99 mmol), it was raised to 35 °C and reacted for 1 hour. The reaction was continued for 1 hour. Additional N,N-diisopropylethylamine (2.32 g, 17.99 mmol) was added, and the reaction was continued for 5 hours. The reaction solution was cooled to 25°C, diluted with ethyl acetate (50 mL), washed with water (2×50 mL), washed with saturated sodium chloride solution (50 mL), dried over anhydrous sodium sulfate, and concentrated. The crude product was separated by column chromatography (mobile phase: petroleum ether: ethyl acetate=10:1-2:1) to obtain Intermediate 2-2. MS: MS(ESI) m/z: 514.1 [M+1] ⁺ .

Step 2: Synthesis of Compounds 2-3

Intermediate 2-2 (2 g, 3.89 mmol), intermediate 1-8a (929.16 mg, 5.84 mmol), tris(dibenzylideneacetone)dipalladium (356.30 mg, 389.09 μmol), 1,1′-dipalladium Naphthalene-2,2'-bisdiphenylphosphine (484.56 mg, 778.19 μmol) and cesium carbonate (2.54 g, 7.78 mmol) were dissolved in dioxane (30 mL) and reacted at 95° C. for 16 hours under nitrogen protection. The reaction solution was filtered, and the mother liquor was concentrated to dryness. The crude product was separated by column chromatography (mobile phase: dichloromethane: methanol=100:1～50:1) to obtain Intermediate 2-3. MS(ESI) m/z: 637.3[M+1] ⁺ .

Step 3: Synthesis of Compounds 2-4

Intermediate 2-3 (2.3 g, 3.61 mmol) was dissolved in tetrahydrofuran (20 mL) and methanol (10 mL), then palladium hydroxide (345.00 mg, 491.33 μmol) was added, and the reaction was carried out at 25°C under hydrogen (15 psi) atmosphere. 18 hours. The reaction solution was filtered and concentrated. The crude product was separated by column chromatography (mobile phase: dichloromethane: methanol=50:1～20:1) to obtain Intermediate 2-4. MS(ESI) m/z: 503.3 [M+1] ⁺ .

Step 4: Synthesis of Compounds 2-5

Intermediate 2-4 (200 mg, 397.91 μmol), Intermediate 2-4a (184.69 mg, 795.82 μmol), tris(dibenzylideneacetone)dipalladium (72.88 mg, 79.58 μmol), 2-bicyclohexylphosphine -2',6'-diisopropoxybiphenyl (74.27 mg, 159.16 μmol) and cesium carbonate (324.12 mg, 994.78 μmol) were dissolved in toluene (4 mL) and reacted at 90° C. for 19 hours under nitrogen protection. The reaction solution was cooled to 25°C, diluted with ethyl acetate (20 mL), insolubles were filtered off, and concentrated. The crude product was separated by column chromatography (mobile phase: dichloromethane: methanol=50:1～20:1) to obtain Intermediate 2-5. LCMS: MS(ESI) m/z: 654.3 [M+1] ⁺ .

Step 5: Synthesis of the formate salt of compound 2

Intermediate 2-5 (129 mg, 197.31 μmol) was dissolved in dichloromethane (2 mL), trifluoroacetic acid (770.00 mg, 6.75 mmol) was added at 25° C., and the reaction was continued for 0.5 hours. The reaction solution was concentrated to dryness, dissolved in dichloromethane (10 mL), sodium bicarbonate solid was added to neutralize excess trifluoroacetic acid, ethyl acetate (20 mL) was added to dilute, insolubles were filtered off, and concentrated. The crude product was separated by preparative HPLC (chromatographic column: Welch Xtimate C18 150*25mm*5μm; mobile phase: [water (formic acid)-acetonitrile]; acetonitrile%: 14%-44%, 6min) to obtain the formate salt of compound 2. MS(ESI) m/z: 554.5[M+1] ⁺ . ¹ H NMR (400MHz, CD ₃ OD) δ 8.23 (br d, J=8.28Hz, 1H), 8.03 (br d, J=7.03Hz, 1H), 7.87 (br d, J=8.03Hz, 1H) ,7.55-7.74(m,3H),5.43-5.66(m,1H),4.40-4.56(m,3H),4.12-4.32(m,3H),3.72-4.05(m,5H),3.50-3.70( m,2H),3.32-3.47(m,4H),3.27-3.31(m,1H),2.62-2.73(m,1H),2.48-2.59(m,1H),2.22-2.42(m,4H), 2.00-2.20 (m, 4H).

Example 3

Step 1: Synthesis of Compound 3-1

Intermediate 2-4 (50 mg, 99.48 μmol) and Intermediate 2-4b (20.80 mg, 99.48 μmol) were dissolved in N,N-dimethylformamide (5 mL) followed by the addition of N,N-diisopropyl Ethylamine (38.57 mg, 298.43 μmol), reacted at 80° C. for 12 hours. Washed with water (5 mL*2), extracted with ethyl acetate (10 mL*2), washed with saturated brine (5 mL), dried over anhydrous sodium sulfate, concentrated under reduced pressure to remove the organic solvent. The crude product was separated by column chromatography (mobile phase: dichloromethane: methanol=50:1～20:1) to obtain Intermediate 3-1. MS(ESI) m/z: 692.3[M+1] ⁺ .

Step 2: Synthesis of Compound 3-2

Intermediate 3-1 (45 mg, 65.06 μmol) was dissolved in methanol (5 mL), purged with nitrogen, palladium on carbon (4.5 mg, 10% pure) was added, purged with nitrogen, and then under an atmosphere of hydrogen (15 psi) , and reacted at 25°C for 4 hours. The palladium/carbon was removed by filtration, and the filtrate was concentrated under reduced pressure to remove the organic solvent to obtain the crude intermediate 3-2. MS(ESI) m/z: 662.3[M+1] ⁺ .

Step 3: Synthesis of the hydrochloride salt of compound 3

Intermediate 3-2 (40 mg, 57.29 μmol) was dissolved in methanol (1 mL), followed by adding hydrogen chloride/dioxane (4 M, 4 mL), and reacted at 25° C. for 4 hours. Directly concentrate under reduced pressure to remove the organic solvent, beat with DCM, and filter to obtain the hydrochloride of compound 3. MS(ESI) m/z: 562.3[M+1] ⁺ . ¹ H NMR (400 MHz, CD ₃ OD) δ 7.67 (s, 1H), 7.53 (d, J=8.0 Hz, 1H), 7.34 (d, J=8.3 Hz, 1H), 5.70-5.55 (m, 1H) ), δ=4.29(br s, 2H), 4.23(s, 2H), 4.02-3.97(m, 3H), 3.82-3.80(m, 3H), 3.51-3.47(m, 2H), 3.29-3.27( m,3H),3.09-3.06(m,3H),2.83-2.74(m,1H),2.74-2.62(m,3H),2.55-2.45(m,2H),2.40-2.35(m,4H), 2.29-2.24 (m, 2H).

Example 4

Step 1: Synthesis of Compounds 4-1a and 4-1b

Intermediate 2-4 (300 mg, 596.87 μmol), 2-dicyclohexylphosphino-2'-(N,N-dimethylamine)-biphenyl (46.98 mg, 119.37 μmol) and intermediate 2-4c ( 478.66 mg, 895.30 μmol) was added to dioxane (3 mL), followed by lithium bistrimethylsilylamide (1 M, 1.19 mL) and tris(dibenzylideneacetone)dipalladium (54.66 mg, 59.69 μmol). After the obtained reaction liquid was replaced with nitrogen, it was placed in an oil bath at 75° C. to stir and react for 6 hours. The reaction solution was spin-dried to obtain crude product. The crude product was purified by column (methanol/dichloromethane=0-6%) to obtain the crude product. The crude product was directly separated by preparative high performance liquid chromatography (chromatographic column: Welch Xtimate C18 100*40mm*3μm; mobile phase: [water (0.075% trifluoroacetic acid)-acetonitrile]; acetonitrile%: 52%-82%, 8min) to obtain Compounds 4-1a and 4-1b. The retention time of 4-1a in LCMS (column: ChromCore 120 C18 3μm 3.0*30mm, mobile phase: [water (0.075% trifluoroacetic acid)-acetonitrile]; acetonitrile %: 30%-90%, 7 min) was 5.102 min , the retention time of 4-1b was 5.038min. 4-1a: MS m/z=887.6[M+H] ⁺ , 4-1b: MS m/z=887.6[M+H] ⁺

Step 2: Synthesis of Compound 4-2a

A methanol solution of hydrogen chloride (4M, 1 mL) was added to Intermediate 4-1a (100 mg, 112.72 μmol), and the reaction was stirred at 20° C. for 2 hours. The reaction solution was spin-dried at 40°C to obtain the crude product of intermediate 4-2a, which was directly used in the next reaction. Retention time of 4-2a in LCMS (column: Merck Chromolith Flash RP-18e 25-3 mm, mobile phase: [water (0.075% trifluoroacetic acid)-acetonitrile]; acetonitrile %: 5%-95%, 1.5 min) is 0.864, MS m/z=743.2[M+H] ⁺ .

Step 3: Synthesis of Compound 4-2b

A methanol solution of hydrogen chloride (4M, 1 mL) was added to Intermediate 4-1b (98 mg, 110.46 μmol), and the reaction was stirred at 20° C. for 2 hours. The reaction solution was spin-dried to obtain the crude product of intermediate 4-2b, and the crude product was directly used in the next reaction. Retention time of 4-2b in LCMS (column: Merck Chromolith Flash RP-18e 25-3 mm, mobile phase: [water (0.075% trifluoroacetic acid)-acetonitrile]; acetonitrile %: 5%-95%, 1.5 min) is 0.970, MS m/z=743.2[M+H] ⁺

Step 4: Synthesis of Compound 4a

Intermediate 4-2a (53 mg, 71.33 μmol) was added to N,N-dimethylformamide (0.5 mL) followed by tetramethylammonium fluoride tetrahydrate (35.35 mg, 213.99 μmol, 3 eq), The obtained reaction liquid was heated to 55°C under nitrogen protection and stirred for 2 hours. The reaction solution was directly separated by high performance liquid chromatography (chromatographic column: Phenomenex C18 80*40mm*3μm; mobile phase: [water (0.05% ammonia water)-acetonitrile]; acetonitrile %: 47%-77%, 8min) to obtain compound 4a. The retention time of 4-a in LCMS (chromatographic column: ChromCore 120 C18 3μm 3.0*30mm, mobile phase: [water (0.075% trifluoroacetic acid)-acetonitrile]; acetonitrile %: 10%-80%, 7min) was 2.422, MS m/z=587.3[M+H] ⁺ .

Step 5: Synthesis of Compound 4b

Intermediate 4-2b (70 mg, 94.21 μmol) was dissolved in N,N-dimethylformamide (1 mL), then tetramethylammonium fluoride tetrahydrate (46.69 mg, 282.63 μmol) was added, and the resulting reaction solution The temperature was raised to 55°C and the reaction was stirred for 2 hours. The reaction solution was spin-dried to obtain crude product. The crude product was dissolved in acetonitrile and separated by high performance liquid chromatography (chromatographic column: Phenomenex C18 80*40mm*3μm; mobile phase: [water (0.05% ammonia water)-acetonitrile]; acetonitrile%: 38%-68%, 8min) to obtain Compound 4b. The retention time of 4-b in LCMS (column: ChromCore 120 C18 3μm 3.0*30mm, mobile phase: [water (0.075% trifluoroacetic acid)-acetonitrile]; acetonitrile %: 10%-80%, 7min) was 2.724, MS m/z=587.3[M+H] ⁺

Example 5

Step 1: Synthesis of Compound 5-1

Intermediate 2-4 (127.13 mg, 397.91 μmol) was dissolved in toluene (5 mL), followed by addition of Intermediate 2-4d (49.55 mg, 79.58 μmol), palladium acetate (8.93 mg, 39.79 μmol) and cesium carbonate (388.94 mg, 1.19 mmol), nitrogen was purged three times, and the reaction was carried out at 100° C. for 5 hours. The insolubles were removed by filtration, and the filtrate was directly concentrated under reduced pressure to obtain crude product 5-1, which was directly used in the next step. MS m/z=741.2[M+H] ⁺

Step 2: Synthesis of Compound 5

Intermediate 5-1 (100 mg, 134.91 μmol) was dissolved in methanol (2 mL), followed by adding hydrogen chloride/methanol solution (4 M, 2 mL), and reacted at 25° C. for 3 hours. The organic solvent was removed by concentration under reduced pressure, and the crude product was separated by preparative HPLC (chromatographic column: Xtimate C18 150*40mm*5μm; mobile phase: [water (0.225% formic acid)-acetonitrile]; acetonitrile%: 10%-50%, 8min) to obtain Compound 5. MS m/z=597.2[M+H] ^{+ .1H} NMR (400MHz, _CD3OD ) 6.77(d, J=2.4Hz, 1H), 6.73(d, J=2.4Hz, 1H), 5.43(d , J=53.2Hz, 1H), 4.95–4.90 (m, 1H), 4.82–4.73 (m, 1H), 4.38–4.15 (m, 3H), 4.09 (s, 2H), 4.01 (s, 2H), 3.60–3.48 (m, 3H), 3.48–3.36 (m, 3H), 3.28–3.17 (m, 3H), 2.87–2.82 (m, 2H), 2.51–2.33 (m, 2H), 2.28–2.04 (m ,8H).

Example 6

Step 1: Synthesis of Compound 6-1a-2

Intermediate 6-1a-1 (20 g, 88.78 mmol) was dissolved in tetrahydrofuran (100 mL) under nitrogen protection, dissolved at 25 °C, then cooled to -78 °C, and lithium diisopropylamide (2M, 57.71 mL), stirred at -78 °C for 0.5 h, then 2-[N,n-bis(trifluoromethanesulfonyl)amino]-5-chloropyridine (46.37 g, 118.07 mmol) was dissolved in tetrahydrofuran ( 50 mL), the solution was then transferred dropwise to the above system, the temperature was raised to 25°C, and the reaction was continued for 2.5 hours. Add 20mL saturated ammonium chloride solution to quench the reaction, spin off most of the organic solvent, wash with water (20mL*2), extract with ethyl acetate (50mL*2), wash with saturated brine (10mL), dry over anhydrous sodium sulfate, spin dry The organic solvent was removed. The crude product was separated by column chromatography (mobile phase: petroleum ether/ethyl acetate=15:1-10:1) to obtain compound 6-1a-2.

Step 2: Synthesis of Compound 6-1a

Intermediate 6-1a-2 (35 g, 97.94 mmol) and bispinacol boronate (29.85 g, 117.53 mmol) were dissolved in dioxane (50 mL), purged with nitrogen, followed by the addition of potassium acetate (28.84 g, 293.83 mmol), [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride (14.33 g, 19.59 mmol), and then purged with nitrogen, and reacted at 80° C. for 12 hours. The insolubles were removed by filtration, concentrated under reduced pressure to remove most of the solvent, then washed with water (50 mL*2), extracted with ethyl acetate (200 mL*2), washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, and concentrated under reduced pressure to remove the solvent. The crude product was separated by column chromatography (mobile phase: petroleum ether/ethyl acetate=10:1-5:1) to obtain compound 6-1a. MS m/z=336.2 [M+H] ⁺ .

Step 3: Synthesis of Compound 6-2

Intermediates 6-1 (3 g, 10.20 mmol) and 6-1a (3.42 g, 10.20 mmol) were dissolved in a mixed solvent of dioxane (15 mL) and water (2 mL), purged with nitrogen, followed by adding carbonic acid in sequence Potassium (4.23g, 30.59mmol), [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride (746.19mg, 1.02mmol), then purged with nitrogen, and reacted at 50°C under nitrogen atmosphere for 5 Hour. The insoluble matter was removed by filtration, and the organic solvent was removed by concentration under reduced pressure. The crude product was separated by column chromatography (mobile phase: petroleum ether/ethyl acetate=10:1-5:1) to obtain compound 6-2. MS m/z=467.2 [M+H] ⁺ .

Step 4: Synthesis of Compound 6-3

Intermediates 6-2 (2 g, 4.28 mmol), 1-8a (1.02 g, 6.42 mmol), palladium acetate (96.15 mg, 428.26 μmol), cesium carbonate (4.19 g, 12.85 mmol), 1,1′-bivalent Naphthalene-2,2'-bisdiphenylphosphine (533.33 mg, 856.53 μmol) was dissolved in toluene (20 mL), purged with nitrogen, and reacted at 100° C. for 12 hours. The insoluble matter was removed by filtration, and the organic solvent was removed by concentration under reduced pressure. The crude product was separated by column chromatography (mobile phase: dichloromethane/methanol=50:1～20:1) to obtain compound 6-3. MS m/z=590.3 [M+H] ⁺ .

Step 5: Synthesis of Compounds 6-4

Intermediate 6-3 (1 g, 1.70 mmol) was dissolved in methanol (10 mL), purged with nitrogen, followed by addition of palladium hydroxide (0.2 g, purity 10%), purged with nitrogen, then purged with hydrogen, under a hydrogen atmosphere The reaction was carried out at 65°C for 24 hours. The solid was removed by filtration, and the filtrate was concentrated under reduced pressure to obtain crude product 6-4. MS m/z=502.2 [M+H] ⁺ .

Step 6: Synthesis of Compounds 6-5

Intermediates 6-4 (300 mg, 598.04 μmol), 2-4d (191.08 mg, 598.04 μmol) were dissolved in dioxane (5 mL), purged with nitrogen, followed by addition of tetrakis(triphenylphosphine)palladium (138.22 mg, 119.61 μmol), 1,1'-binaphthyl-2,2'-bisdiphenylphosphine (148.95 mg, 239.22 μmol) and cesium carbonate (584.56 mg, 1.79 mmol), purged with nitrogen, and then warmed to 90 ℃, the reaction was carried out for 12 hours. The insoluble matter was removed by filtration, and the organic solvent was removed by concentration under reduced pressure. The crude product was separated by preparative HPLC (chromatographic column: Phenomenex C18 80*40mm*3μm; mobile phase: [water (0.05% ammonia water)-acetonitrile]; acetonitrile%: 74%-100%, 8min) to obtain compound 6-5. MS m/z=740.1 [M+H] ⁺ .

Step 7: Synthesis of Compounds 6a and 6b

Intermediate 6-5 (200 mg, 216.15 μmol) was dissolved in methanol (5 mL), then hydrogen chloride/methanol (4 M, 1.08 mL) was added, and the reaction was carried out at 25° C. for 4 hours. The organic solvent was removed by concentration under reduced pressure, and the crude product was separated by SFC (chromatographic column: DAICEL CHIRALPAK IG (250mm*30mm, 10μm); mobile phase: [0.1% ammonia water-ethanol]; % ethanol: 50%-50%, 8min) to obtain the compound 6a and 6b. The retention time of 6a in LCMS (column: ChromCore 120 C18 3μm 3.0*30mm, mobile phase: [water (0.075% trifluoroacetic acid)-acetonitrile]; acetonitrile %: 10%-80%, 7min) was 2.810, MS m /z=596.4[M+H] ⁺ , the retention time of 6b is 3.035, MS m/z=596.3[M+H] ⁺ , the NOESY spectrum identifies that the hydrogen at the 4-position of piperidine in 6b is related to the hydrogen on the bridge ring. NOE correlation, the hydrogen at position 4 of 6a piperidine and the hydrogen on the bridge ring have no NOE correlation, so the two structures can be distinguished.

Experimental example 1. KRAS ^G12D inhibitory activity test

1. Experimental purpose:

Through the TR-FRET method, compounds that can effectively inhibit the binding of KRAS ^G12D to GTP were screened.

2. Consumables and instruments:

Table 3. Consumables and Instruments

3. Reagent preparation:

a. Storage reagents:

1) KRAS Nucleotide Exchange Buffer

Take 20mL of 1000mM HEPES, 20mL of 500mM EDTA, 10mL of 5M sodium chloride, 100% 0.1mL of Tween 20, and 949.9mL of water to prepare a 1L solution, sterilized by filtration, and stored at 4°C.

2) KRAS assay buffer

Take 20mL of 1000mM HEPES, 10mL of 1000mM magnesium chloride, 30mL of 5M sodium chloride, 100% 0.05mL Tween 20, and 939.95mL of water to prepare a 1L solution, sterilized by filtration, and stored at 4°C.

3) KRAS/Bodipy GDP/Tb-SA mixture

Take 9.5μL of 95μM KRAS ^G12D protein, mixed with 440.5μL of KRAS nucleotide exchange buffer, and incubated at room temperature for 1 hour, then mixed with 8.4μL of 17.9μM Tb-SA, 1.8μL of 5mM Bodipy GDP, and 9539.8μL of KRAS experimental buffer. 1L of the solution, after mixing, let it stand at room temperature for 6 hours, and store it at -80°C.

b. Experimental reagents:

1) KRAS enzyme solution

Take 73.3 μL KRAS/Bodipy GDP/Tb-SA mixture and 2126.7 μL KRAS experimental buffer to prepare a 2200 μL solution.

2) SOS/GTP mixture

Take 1.59 μL of 166 μM SOS protein, 198 μL of 100 mM GTP, and 2000.41 μL of KRAS experimental buffer to prepare a 2200 μL solution.

4. Experimental process:

1) The concentration of the control compound stock solution is 1 mM, and the concentration of the test compound stock solution is 10 mM. Transfer 9 μL of control compound and test compound to 384-LDV plate;

2) Use Bravo to make 10:3 dilutions of the compounds on the LDV plate;

3) Use ECHO to transfer 9nL of the compound on the LDV plate to the experimental plate;

4) Add 3 μL of 3nM Kras/0.5nM TB-SA/30nM BodipyGDP mixture and 3 μL of Ras buffer to each well of the experimental plate in turn using a Dragonfly automatic sampler, and centrifuge the experimental plate for 1 minute at 1000rpm/min;

5) Incubate the experimental plate for 1 hour at room temperature;

6) Add 3 μL of 120nM SOS/9mM GTP mixture to each well of the experimental plate using a Dragonfly automatic sampler, and centrifuge the experimental plate for 1 minute at 1000rpm/min;

7) Incubate the experimental plate for 1 hour at room temperature;

8) Use Envision to read the plate and record the data;

9) Use Excel and Xlfit for data analysis to calculate the IC ₅₀ of the test compound.

5. Experimental results:

The results are shown in Table 4.

Table 4 _IC50 values of compounds for KRAS ^G12D enzyme inhibition

Compound number	KRAS G12D IC 50 (nM)
Compound 1	43
Formate salt of compound 2	149

6. Experimental conclusion:

The compounds of the present invention have a significant inhibitory effect on KRAS ^G12D enzyme.

Claims (20)

1. The compound represented by formula (II) or a pharmaceutically acceptable salt thereof

   

   in,

   

   is selected from single bond or double bond;

   T ₁ is selected from CR ₇ R ₈ , NR ₉ and O;

   when

   

   is selected from single bond, T is selected from _CH and N;

   when

   

   is selected from _double bonds, and T is selected from C;

   L ₁ is selected from -CH ₂ - and a bond;

   R ₁ , R ₂ , R ₃ , R ₄ and R ₅ are each independently selected from H and C _1-3 alkyl optionally substituted with ₁ , 2 or 3 _Ra ;

   R ₆ is selected from C _6-10 aryl and 5-10 membered heteroaryl, the C _6-10 aryl and 5-10 membered heteroaryl are optionally surrounded by 1, 2, 3, 4 or 5 R _b replace;

   R ₇ and R ₈ are each independently selected from H, CH ₃ and NH ₂ ;

   R ₉ is selected from H and CH ₃ ;

   R ₁₀ is selected from H, C _1-3 alkyl, C _1-3 alkoxy and cyclopropyl, the C _1-3 alkyl, C _1-3 alkoxy and cyclopropyl are optionally replaced by 1, 2 or 3 R _c substitutions;

   R ₁₁ is selected from 4-8 membered heterocycloalkyl and

   

   The 4-8 membered heterocycloalkyl and

   

   optionally substituted with 1, 2 or 3 _Rd ;

   Structural units

   

   selected from 5-6 membered heterocycloalkenyl;

   requirement is,

   1) R ₁ and R ₂ form a ring with the connected atoms, making the structural unit

   

   form

   

   2) Alternatively, R ₁ and R ₂ form a ring with the connected atoms, making the structural unit

   

   form

   

   3) Alternatively, R ₁ and R ₄ form a ring with the connected atoms, making the structural unit

   

   form

   

   4) Alternatively, R ₄ and R ₅ form a ring with the connected atoms, making the structural unit

   

   form

   

   5) Alternatively, R ₂ and R ₇ form tetrahydropyrrolidinyl with the attached atoms;

   6) Alternatively, R ₂ and R ₃ and the connected atom form a C _3-5 membered cycloalkyl;

   7) Alternatively, R ₇ and R ₈ form a 4-5 membered heterocycloalkyl with the attached atom;

   m is selected from 0, 1 or 2;

   n is selected from 0, 1 or 2;

   p is selected from 0, 1 or 2;

   q is selected from 1, 2 or 3;

   r is selected from 1 or 2;

   s is selected from 1, 2 or 3;

   each R _a is independently selected from F, Cl, Br and I;

   Each R _b is independently selected from F, Cl, Br, I, OH, NH ₂ , CN, C _1-3 alkyl, C _1-3 alkoxy, C _2-3 alkynyl, C _2-3 alkene and C _3-5 cycloalkyl, the C _1-3 alkyl, C _1-3 alkoxy, C _2-3 alkynyl, C _2-3 alkenyl and C _3-5 cycloalkyl are optional replaced by 1, 2 or 3 Rs;

   each R _c is independently selected from F, Cl, Br and I;

   Each R _d is independently selected from H, F, Cl, Br, OH, CN, C _1-3 alkyl, C _1-3 alkoxy and -C _1-3 alkyl-OC(=O)-C _1-3 alkylamino;

   Each R is independently selected from F, Cl and Br.
2. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein the structural unit

   

   selected from

   

   
3. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein the structural unit

   

   selected from

   
4. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein the structural unit

   

   selected from

   

   
5. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein the structural unit

   

   selected from

   
6. The compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein R ₂ and R ₇ form with the attached atom

   
7. The compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein R ₂ and R ₃ form with the atoms to which they are attached

   
8. The compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein R ₇ and R ₈ form with the attached atom

   
9. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein the structural unit

   

   selected from

   

   
10. The compound of claim 1 or a pharmaceutically acceptable salt thereof, wherein each R _b is independently selected from F, Cl, Br, I, OH, NH ₂ , CN, CH ₃ , CH ₂ CH ₃ , OCH ₃ , OCH ₂ CH ₃ , -CH=CH ₂ , -CH ₂ -CH=CH ₂ , -C≡CH and cyclopropyl, the CH ₃ , CH ₂ CH ₃ , OCH ₃ , OCH ₂ CH ₃ , -CH = _CH2 , -CH2-CH= _CH2 , -CH= _CH2 and cyclopropyl are optionally substituted with 1, ₂ or 3 Rs.
11. The compound of claim 10 or a pharmaceutically acceptable salt thereof, wherein each R _b is independently selected from F, Cl, OH, NH ₂ , CN, CH ₃ , CF ₃ , CH ₂ CH ₃ , -C≡ CH and cyclopropyl.
12. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein R ₆ is selected from phenyl, pyridyl, naphthyl, indolyl and indazolyl, the phenyl, pyridyl, naphthyl, Indolyl and indazolyl are optionally substituted with 1, 2, 3, 4 or 5 R _b .
13. The compound according to claim 12 or a pharmaceutically acceptable salt thereof, wherein R ₆ is selected from

    

    
14. The compound according to any one of claims 1, 12 or 13, or a pharmaceutically acceptable salt thereof, wherein R ₆ is selected from

    

    
15. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein R ₁₀ is selected from H, CH ₃ , OCH ₃ and cyclopropyl, wherein said CH ₃ , OCH ₃ and cyclopropyl are optionally 2 or 3 R _c substitutions.
16. The compound of claim 1 or 15, or a pharmaceutically acceptable salt thereof, wherein R ₁₀ is selected from H and CH ₃ .
17. The compound of claim 1 or a pharmaceutically acceptable salt thereof, wherein each R _d is independently selected from H, F, Cl, Br, OH, CN, CH ₃ , CH ₂ CF ₃ , OCH ₃ and

    
18. The compound of claim 1 or a pharmaceutically acceptable salt thereof, wherein R ₁₁ is selected from tetrahydropyrrolyl and hexahydro-1H-pyrrolizinyl, said tetrahydropyrrolyl and hexahydro-1H-pyrrole Rizinyl is substituted with 1, 2 or 3 _Rd .
19. The compound according to any one of claims 1, 17 or 18, or a pharmaceutically acceptable salt thereof, wherein R ₁₁ is selected from

    

    
20. The compound represented by the following formula or its pharmaceutically acceptable salt, its compound is selected from,

    

    

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