CN112480116A

CN112480116A - PKB inhibitors

Info

Publication number: CN112480116A
Application number: CN201910858374.XA
Authority: CN
Inventors: 马昌友; 张林林; 吴有智; 吴舰; 徐丹; 朱春霞; 田舟山
Original assignee: Chia Tai Tianqing Pharmaceutical Group Co Ltd
Current assignee: Chia Tai Tianqing Pharmaceutical Group Co Ltd
Priority date: 2019-09-11
Filing date: 2019-09-11
Publication date: 2021-03-12
Anticipated expiration: 2039-09-11
Also published as: CN112480116B

Abstract

The invention provides a PKB inhibitor, in particular to a compound shown as a formula I or a pharmaceutically acceptable salt thereof, and also provides a preparation method thereof.

Description

PKB inhibitors

Technical Field

The invention belongs to the field of pharmaceutical chemistry, and particularly relates to a PKB inhibitor, a preparation method and a medical application thereof.

Background

The PI3K/AKT/mTOR pathway composed of phosphatidylinositol 3-kinase (PI3K) and its downstream protein AKT (also called protein kinase B, PKB) and mammalian target of rapamycin (mTOR) is used as a very important signal transduction pathway in cells, and plays an extremely important biological function in the processes of cell growth, survival, proliferation, apoptosis, angiogenesis, autophagy and the like. Abnormal activation of this pathway can lead to a range of diseases including cancer, neuropathy, autoimmune diseases and haemo-lymphatic diseases.

AKT has three subtypes: AKT1, AKT2, and AKT 3. As a typical protein kinase, each subtype consists of an amino-terminal PH domain (Pleckstrin homology domain), a kinase domain that binds ATP centrally, and a carboxy-terminal regulatory domain. The amino acid sequences of about 80% of the 3 subtypes are homologous and vary greatly only at the junction of the PH domain and the kinase domain.

Currently, the targeted drugs for the PI3K/AKT/mTOR signaling pathway are mainly PI3K inhibitors and mTOR inhibitors, while AKT is in the core part of the signaling pathway. Inhibition of AKT activity can avoid both the severe side effects caused by inhibition of upstream PI3K and the effects of negative feedback mechanisms caused by inhibition of downstream mTOR. Therefore, finding potent and selective AKT inhibitors is an important direction for the development of current tumor-targeted drugs.

Disclosure of Invention

In one aspect, the invention provides a compound of formula I or a pharmaceutically acceptable salt thereof,

wherein:

R¹and R²Each independently selected from H or C1-C6 alkyl;

m is selected from 0 or 1;

a is optionally substituted by 1-5R³Substituted phenyl, wherein each R is³Each independently selected from halogen or C1-C6 alkyl, wherein C1-C6 alkyl may be optionally substituted with halogen;

is selected from

G is selected from 5-6 membered heteroaryl or phenyl, wherein said heteroaryl or phenyl is optionally substituted with R⁴Is substituted, and R⁴Selected from halogen or C1-C6 alkyl;

l is

A group wherein:

Y¹is selected from CY¹¹Y¹²Or NY¹³，Y²Selected from O, CY²¹Y²²Or a bond, and satisfies when Y²Selected from O, Y¹Is selected from CY¹¹Y¹²And, in addition,

Y¹¹and Y²¹Each independently selected from H, OH, halogen or C1-C6 alkyl, wherein C1-C6 alkyl may be optionally substituted with halogen, OH, C1-C3 alkoxy or CN,

Y¹²and Y²²Each independently selected from H or C1-C6 alkyl;

Y¹³selected from H or C1-C6 alkyl, wherein C1-C6 alkyl may be optionally substituted with halogen, OH, C1-C3 alkoxy or CN.

In some embodiments, R¹And R²Each independently selected from H, methyl, ethyl or isopropyl; preferably, R¹Selected from H, R²Selected from H, methyl, ethyl or isopropyl; more preferably, R¹Selected from H, R²Selected from H or isopropyl; most preferably, R¹Selected from H, R²Is selected from H.

In some embodiments, m is selected from 1.

In some embodiments, A is substituted with 1 or 2R³A substituted phenyl group.

In some embodiments, a is selected from the group consisting of:

in some embodiments, R³Independently selected from F, Cl, Br, I, CH₃、C₂H₅Or CF₃(ii) a In some typical embodiments, R³Selected from F, Cl or CF₃(ii) a In some more typical embodiments, R³Selected from F or Cl; in some of the most typical embodiments, R³Is selected from F.

In some embodiments, a is selected from the group consisting of:

in some typical embodiments, a is selected from the group consisting of:

in some typical embodiments, a is selected from the group consisting of:

in some more typical embodiments, A is selected from

In some embodiments of the present invention, the substrate is,

is selected from

In some embodiments, G is selected from: a five-membered heteroaryl, pyridyl or phenyl group, wherein the five-membered heteroaryl, pyridyl or phenyl group is optionally substituted with R⁴Is substituted, and R⁴Selected from halogensOr C1-C6 alkyl.

In some embodiments, R⁴Is selected from CH₃F or Cl; in some typical embodiments, R⁴Is selected from CH₃Or F.

In some embodiments, G is selected from optionally substituted with R⁴Substituted of the following groups:

wherein,

represents the position where G is attached to Q by a chemical bond,

represents the position at which G is attached to L by a chemical bond;

R⁴as defined above.

In some typical embodiments, G is selected from optionally substituted with R⁴Substituted of the following groups:

wherein R is⁴As defined above.

In some more typical embodiments, G is selected from optionally substituted with R⁴Substituted of the following groups:

wherein R is⁴As defined above.

In some more typical embodiments, G is selected from the following groups:

in some more typical embodiments, G is selected from the following groups:

in some more typical embodiments, G is selected from the following groups:

in some embodiments, Y is¹Is selected from CY¹¹Y¹²Wherein Y is¹¹Selected from H, OH or C1-C6 alkyl, wherein C1-C6 alkyl may be optionally substituted with F; in some exemplary embodiments, Y¹¹Selected from H, CH₃Or CF₃(ii) a In some more typical embodiments, Y¹¹Is selected from CH₃Or CF₃(ii) a In some of the most typical embodiments, Y¹¹Is selected from CH₃。

In some embodiments, Y is¹²Is selected from H or CH₃。

In some embodiments, Y is¹Is selected from NY¹³Wherein Y is¹³Selected from H, CH₃、C₂H₅、CH₂CH₂CH₃、CHCH₃CH₃、CH₂OH、C₂H₄OH、C₃H₆OH、CH₂OCH₃、CH₂OC₂H₅、C₂H₄OCH₃、CH₂CN or C₂H₄CN; preferably, Y is¹³Is selected from CH₃、C₂H₄OH or CH₂CN; in some more typical embodiments, Y¹³Is selected from CH₃。

In some embodiments, Y is²Selected from O, CY²¹Y²²Or a bond, wherein Y²¹And Y²²Are all H; in some exemplary embodiments, Y²Is selected from CH₂Or a key.

In some embodiments, L is selected from the group consisting of:

in some typical embodiments, G is selected from the following groups:

in some more typical embodiments, G is selected from the following groups:

in another aspect, the invention provides a compound of formula II or a pharmaceutically acceptable salt thereof,

wherein, A, G, L, R¹And R²Are as defined for compounds of formula I.

In another aspect, the invention provides a compound of formula III-1 or a pharmaceutically acceptable salt thereof,

wherein, A, R¹、R²And G is as defined for the compound of formula I, R³¹A compound Y of the same formula I¹¹，R³²A compound Y of the same formula I¹²。

In another aspect, the invention provides a compound of formula III-2 or a pharmaceutically acceptable salt thereof,

wherein, A, R¹、R²And G is as defined for the compound of formula I, R³³A compound Y of the same formula I¹³The definition of (1).

In another aspect, the invention provides a compound of formula III-3 or a pharmaceutically acceptable salt thereof,

wherein, A, R¹、R²And G is as defined for the compound of formula I, R³⁴A compound Y of the same formula I¹¹，R³⁵A compound Y of the same formula I¹²。

In another aspect, the invention provides a compound of formula III-4 or a pharmaceutically acceptable salt thereof,

wherein, A, R¹、R²And G is as defined for the compound of formula I, R³⁶A compound Y of the same formula I¹³The definition of (1).

In another aspect, the present invention provides the following compounds or pharmaceutically acceptable salts thereof:

in another aspect, the present invention provides a method for the prevention and/or treatment of AKT protein kinase mediated diseases or conditions, comprising administering to a subject in need thereof a compound of formula I of the present invention or a pharmaceutically acceptable salt thereof.

In another aspect, the present invention also provides a compound of formula I of the present invention or a pharmaceutically acceptable salt thereof for use in the prevention and/or treatment of AKT protein kinase mediated diseases or conditions.

In yet another aspect, the present invention provides a process for preparing compounds of formula I, including, but not limited to, the following synthetic schemes:

wherein G, A is as previously defined; y is¹、Y²As defined above or a derivative substituted with a protecting group; x is a leaving group such as halogen; p¹Is C1-C6 alkyl, preferably methyl, ethyl or isopropyl; p²、P³Selected from H or together with the atoms to which they are attached form a boronic ester; "" indicates that the compound may be in optically pure form or a mixture of two or more optically pure forms.

A compound of formula 1-1 is reacted with a compound of formula 1-2 in the presence of a base (e.g., cesium carbonate), a catalyst (e.g., Pd (dppf) Cl₂) And a solvent (e.g., 1, 4-dioxane) to produce a compound of formula 1-3; preparing a compound shown in a formula 1-4 by a hydrolysis reaction of a compound shown in a formula 1-3; reacting a compound of formulae 1-4 with a compound of formulae 1-5 in the presence of a base (e.g., N-diisopropylethylamine), a condensing agent (e.g., BOP), and a solvent (e.g., DMF) to produce a compound of formulae 1-6; the compound of formula 1-6 is reacted under hydrazine hydrate conditions to produce the compound of formula 1-7.

Detailed Description

Correlation definition

Unless specifically stated otherwise, the following terms used in the specification and claims have the following meanings:

the term "compound" as used herein includes all stereoisomers and tautomers.

The compounds of the invention may be asymmetric, e.g., having one or more stereoisomers. Unless otherwise indicated, all stereoisomers include, for example, enantiomers and diastereomers. The compounds of the invention containing asymmetric carbon atoms can be isolated in optically active pure form or in racemic form. The optically active pure form can be resolved from a racemic mixture or synthesized by using chiral starting materials or chiral reagents. Racemates, diastereomers, enantiomers are included within the scope of the present invention.

The compounds of the invention also include tautomeric forms. Tautomeric forms result from the exchange of one single bond with an adjacent double bond and the concomitant migration of one proton.

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

The term "selected from" means that the subsequently described event or circumstance must occur.

Numerical ranges herein refer to each integer in the given range. For example, "C1-C6" means that the group may have 1 carbon atom, 2 carbon atoms, 3 carbon atoms, 4 carbon atoms, 5 carbon atoms, or 6 carbon atoms.

The term "substituted" means that any one or more hydrogen atoms on a particular atom or group is replaced with a substituent, so long as the valency of the particular atom or group is normal and the substituted compound is stable. Unless otherwise specified, the kind and number of substituents may be arbitrary on the basis that they can be chemically achieved.

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 1-5R, the group may optionally be substituted with up to 5R, 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.

The term "alkyl" refers to a saturated aliphatic hydrocarbon group, including straight or branched chain saturated hydrocarbon groups having the indicated number of carbon atoms. For example, the term "C1-C6 alkyl" includes C1 alkyl, C2 alkyl, C3 alkyl, C4 alkyl, C5 alkyl, C6 alkyl, examples include, but are not limited to, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, n-pentyl, 2-pentyl, 3-pentyl, n-hexyl, 2-hexyl, 3-hexyl and the like. It may be divalent, e.g. methylene, ethylene.

The term "alkoxy" refers to a group having the structure-O-alkyl, alkyl being a saturated monovalent hydrocarbon group including straight or branched chains. For example, "C1-C3 alkoxy" includes-OCH₃、-OC₂H₅、-OC₂H₄CH₃and-OCH (CH)₃)₂。

The term "5-6 membered heteroaryl" refers to an aryl group comprising at least one 5-or 6-membered ring independently selected from nitrogen, oxygen and sulfur heteroatoms. Examples of heteroaryl groups include, but are not limited to, imidazolyl, pyrazolyl, thiazolyl, isothiazolyl, furyl, pyrrolyl, thienyl, pyridyl, oxazolyl, pyrimidinyl, pyridazinyl, pyrazinyl, 1,2, 4-triazinyl, 1,3, 5-triazinyl, 1,2, 3-triazinyl.

The term "member" refers to the number of backbone atoms that make up the ring. For example, "5-6 membered" means that the number of backbone atoms constituting the ring is 5 or 6.

All refer to the chemical bond connection part, and both have the same chemical meaning, if no special description,

and

only to distinguish the position or order of the connections.

The term "pharmaceutically acceptable salt" refers to salts that retain the biological potency of the free acid and base of the particular compound without biological adverse effects. Such as acid (including organic and inorganic) addition salts or base addition salts (including organic and inorganic bases).

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

M：mol/L

mM：mmol/L

nM：nmol/L

h: hour(s)

min: is divided into

¹H NMR: hydrogen spectrum of nuclear magnetic resonance

LC/MS: liquid phase/mass spectrometry combination

The DIAD: diisopropyl azodicarboxylate

DCM: methylene dichloride

BOP: benzotriazol-1-yloxytris (dimethylamino) phosphonium hexafluorophosphates

Boc: tert-butyloxycarbonyl radical

The preparation method comprises the following steps:

the following more specifically describes the preparation of the compounds of the present invention, but these specific preparation methods do not set any limit to the scope of the present invention. In addition, reaction conditions such as reactants, solvents, bases, amounts of compounds used, reaction temperature, reaction time and the like are not limited to the following examples.

The compounds of the present invention may also be conveniently prepared by optionally combining various synthetic methods described in the present specification or known in the art, and such combinations may be readily carried out by those skilled in the art.

Preparation example 1

Synthesis of methyl 5- (4,4,5, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) thiophene-3-carboxylate

2.0g of 5-bromothiophene-3-carboxylic acid methyl ester, 3.0g of pinacol diboron ester,0.34g of [1,1' -bis (diphenylphosphino) ferrocene]Dissolving palladium dichloride and 1.8g of potassium acetate in 20mL of dioxane, heating to reflux under the protection of nitrogen, stopping heating after 4 hours, decompressing and evaporating reaction liquid, and performing column chromatography to obtain 1.6g of white solid.¹H NMR(400MHz,CDCl₃)δ8.33(d,J＝1.1Hz,1H),8.04(d,J＝1.1Hz,1H),3.86(s,3H),1.35(s,12H).

Preparation example 2

Synthesis of 4-bromo-5-methylthiophene-2-carboxylic acid

Dissolving 2.2g of liquid bromine in 5mL of acetic acid, dropwise adding the solution into 30mL of acetic acid solution containing 2.0g of 5-methylthiophene-2-formic acid and 0.46g of ferric trichloride at room temperature, stirring at room temperature for 5 hours, pouring ice water, separating out solids, carrying out suction filtration, washing a filter cake with the ice water, and drying to obtain 2.66g of yellow solids. LC/MS (ESI +):221.0(M + H)

Preparation example 3

Synthesis of 4-bromo-5-methylthiophene-2-carboxylic acid methyl ester

At room temperature, 2.65g of 4-bromo-5-methylthiophene-2-carboxylic acid is dissolved in 50mL of methanol, 3mL of concentrated sulfuric acid is added, the temperature is heated to 50 ℃, the reaction solution is cooled to room temperature after 12 hours, the reaction solution is poured into ice water, the pH value is adjusted to 10-11 by using a 2M NaOH solution, DCM is used for extraction, an organic phase is taken, dried by using anhydrous sodium sulfate and evaporated to dryness under reduced pressure, and 2.7g of brown solid is obtained. LC/MS (ESI +):234.9(M + H).¹H NMR(400MHz,DMSO-d₆)δ7.71(d,J＝1.4Hz,1H),3.82(s,3H),2.43(s,3H).

Preparation example 4

Synthesis of methyl 5-methyl-4- (4,4,5, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) thiophene-2-carboxylate

2.0g of methyl 4-bromo-5-methylthiophene-2-carboxylate and 2.8g of pinacol diboron, 0.3g of [1,1' -bis (diphenylphosphino) ferrocene]Dissolving palladium dichloride and 1.7g of potassium acetate in 30ml of dioxane, heating to reflux under the protection of nitrogen, stopping heating after 4 hours, decompressing and evaporating reaction liquid, and performing column chromatography to obtain 1.9g of white solid. LC/MS (ESI +):283.1(M + H).¹H NMR(400MHz,CDCl₃)δ7.95(s,1H),3.86(s,3H),2.72(s,3H),1.34(s,12H).

Preparation example 5

Synthesis of methyl 2-methyl-5- (4,4,5, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) thiophene-3-carboxylate

0.9g of methyl 5-bromo-2-methylthiophene-3-carboxylate, 1.46g of pinacol diboron, 0.28g of [1,1' -bis (diphenylphosphino) ferrocene]Dissolving palladium dichloride and 0.74g of potassium acetate in 20mL of dioxane, heating to reflux under the protection of nitrogen, stopping heating after 4 hours, decompressing and evaporating reaction liquid, and performing column chromatography to obtain 0.65g of white solid.¹H NMR(400MHz,DMSO-d₆)δ7.72(s,1H),3.78(s,3H),2.71(s,3H),1.28(s,12H).

Preparation example 6

Synthesis of methyl 3-fluoro-4- (4,4,5, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) thiophene-2-carboxylate

4-bromo-3-fluorothiophene-2-carboxylic acid methyl ester (2.0g) and pinacol diboron (2.8g), palladium [1,1' -bis (diphenylphosphino) ferrocene ] dichloride (0.3g) and potassium acetate (1.7g) are dissolved in dioxane, heated to reflux under the protection of nitrogen, the heating is stopped after 4h, and the reaction solution is subjected to reduced pressure, evaporation and distillation and then subjected to column chromatography to obtain a white solid (1.7 g). LC/MS (ESI +):287.1(M + H).

Preparation example 7

Synthesis of methyl 3-methyl-5- (4,4,5, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) thiophene-2-carboxylate

4-bromo-3-methylthiophene-2-carboxylic acid methyl ester (2.1g) and pinacol diboron (2.9g), palladium [1,1' -bis (diphenylphosphino) ferrocene ] dichloride (0.3g) and potassium acetate (1.7g) are dissolved in dioxane, heated to reflux under the protection of nitrogen, the heating is stopped after 4h, the reaction solution is decompressed and evaporated to dryness, and then column chromatography is carried out to obtain a white solid (2.0 g). LC/MS (ESI +):283.1(M + H).

Preparation example 8

Synthesis of (R) -4-chloro-5-methyl-5, 8-dihydropyrido [2,3-d ] pyrimidin-7 (6H) -one

Reaction conditions are as follows: a) ethyl crotonate, sodium methoxide methanol solution (30% wt), methanol; b) disodium hydrogen phosphate, deionized water, hydrochloric acid, a lipase (candida rugosa), and sodium hydroxide; c) formamidine acetate, sodium methoxide, methanol; d) phosphorus oxychloride, diisopropylethylamine, acetonitrile; e) ammonia (25-28% wt);

a) 2-methylpropane-1, 1, 3-tricarboxylic acid trimethyl ester

Under the protection of nitrogen, a methanol solution of sodium methoxide (30% wt, 50.32g) was added to methanol (900mL) at 20 ℃, then the temperature was raised to 70 ℃, dimethyl malonate (461.12g) and ethyl crotonate (349.46g) were uniformly mixed, and the mixture was added dropwise to the methanol solution of sodium methoxide and reacted at 70 ℃ for 3 hours. After completion of the reaction, the solvent was distilled off under reduced pressure, ethyl acetate (1L) was added, the pH was adjusted to 7 to 8 with 4M hydrochloric acid, and then 500mL of water was added, followed by liquid separation, and the organic phase was distilled off under reduced pressure to obtain 777.68g of a yellow liquid.¹H NMR(400MHz，DMSO-d₆)δ(ppm)3.67(s，3H)，3.65(s，3H)，3.59(s，3H)，3.56(d，J＝6.8Hz，1H)，2.45-2.58(m，2H)，2.23-2.29(m，1H)，0.93(d，J＝6.8Hz，3H)。

b) (R) -2-methylpropane-1, 1, 3-tricarboxylic acid trimethyl ester

Dissolving disodium hydrogen phosphate (4.5g) in 1.5L deionized water at 25 deg.C, adjusting pH to 7.05 with 2N hydrochloric acid, adding 2-methylpropane-1, 1, 3-tricarboxylic acid trimethyl ester (150.46g) and lipase (Candida rugosa, 40g added in 6 days), adjusting pH to 7.0-7.6 with 2N sodium hydroxide solution, reacting at 35 deg.C for 6 days, and detecting chiral ee%>98% of chiral detection conditions (chiral column AS, n-hexane: ethanol ═ 9:1, 0.2% dimethylethylamine by volume). The reaction solution was cooled to 10 ℃, pH was adjusted to 3-4 with 3M hydrochloric acid, 500mL of ethyl acetate was added, suction filtration was performed, the filter cake was washed with ethyl acetate (600mL), liquid separation was performed, saturated aqueous sodium bicarbonate solution (100mL) was added, liquid separation was performed, and the organic phase was concentrated to obtain 26.89g of a pale yellow liquid.¹H NMR(400MHz，CDCl₃)δ(ppm)3.74(s，6H)，3.68(s，3H)，3.46(d，J＝7.2Hz，1H)，2.71-2.79(m，1H)，2.54(dd，J＝15.6、4.8Hz，1H)，2.32(dd，J＝16.0、8.4Hz，1H)，1.06(d，J＝6.8Hz，3H)。

c) (R) -3- (4, 6-dihydroxypyrimidin-5-yl) butanoic acid methyl ester

Under the protection of nitrogen, formamidine acetate (11.33g) is dissolved in methanol (200mL) at 20 ℃, cooled to 0 ℃, sodium methoxide methanol solution (30%, 55.62g) is added dropwise, reaction is carried out at 0 ℃ for 60min, methanol (60mL) solution of (R) -2-methylpropane-1, 1, 3-tricarboxylic acid trimethyl ester (24.07g) is added dropwise, and the temperature is naturally raised to 20 ℃ for reaction for 10 hours. After completion of the reaction, the reaction solution was cooled to 0 ℃, 3N hydrochloric acid was added to adjust pH to 5 to 6, the solvent was evaporated under reduced pressure, followed by cooling to 0 ℃, 3N hydrochloric acid was added to adjust pH to 3, a solid was precipitated, the solid was collected by suction filtration, the filter cake was washed with ice water (100mL), and the filter cake was dried under vacuum to give 18.79g of a white solid which was used directly in the next step.

d) (R) -3- (4, 6-dichloropyrimidin-5-yl) butanoic acid methyl ester

Under the protection of nitrogen, methyl (R) -3- (4, 6-dihydroxypyrimidin-5-yl) butyrate (14.63g) is dispersed in acetonitrile (70mL) at 22 ℃, phosphorus oxychloride (26.42g) and diisopropylethylamine (12.51g) are added dropwise, the system is obvious in heat release, then the temperature is raised to 60 ℃, the solid is gradually dissolved and cleared, and the reaction is continued for 18 hours. After completion of the reaction, the reaction solution was cooled to 0 ℃, 100mL of ethyl acetate was added, the pH was adjusted to 7 to 8 with a saturated sodium bicarbonate solution, extraction was performed with ethyl acetate (50 mL. times.3), and the organic phase was distilled off under reduced pressure to obtain 13.89g of a yellow solid which was used in the next step as it was.

e) (R) -4-chloro-5-methyl-5, 8-dihydropyrido [2,3-d ] pyrimidin-7 (6H) -one

Methyl (R) -3- (4, 6-dichloropyrimidin-5-yl) butyrate (13.89g) and ammonia (25-28% wt, 70mL) were added to a 100mL autoclave at 20 ℃ and the temperature was raised to 50 ℃ to react for 18 hours. After completion of the reaction, the reaction mixture was cooled to 0 ℃, filtered, and the filter cake was slurried with 30mL of (petroleum ether: ethyl acetate: 10:1, volume ratio) to obtain 7.32g of a pale yellow solid. LC-MS (ESI) M/z:198(M + H).¹H NMR(300MHz，CDCl₃)δ(ppm)1.30(d，J＝7.2Hz，3H)，2.65-2.69(m，1H)，2.86-2.92(m，1H)，3.47-3.54(m，1H)，8.64(s，1H)，10.10(s，1H)。

Preparation example 9

Synthesis of 4-chloro-5-methyl-5, 8-dihydropteridine-7 (6H) -ketone

Reaction conditions are as follows: a) ethyl bromoacetate, sodium hydride, tetrabutylammonium iodide, tetrahydrofuran; b)2, 4-dimethoxybenzylamine, triethylamine and isopropanol; c) methyl iodide, sodium hydride, N dimethylformamide; d) trifluoroacetic acid;

a)2- ((4, 6-dichloropyrimidin-5-yl) amino) acetic acid ethyl ester

To a solution of 4, 6-dichloro-5-aminopyrimidine (10.0g) in tetrahydrofuran (100mL) at 0 deg.C was added sodium hydride (2.93 g). The reaction mixture was stirred for 2 min. After naturally warming to 20 ℃, ethyl bromoacetate (12.22g) was added, and tetrabutylammonium iodide (27.03g) was further added. The reaction mixture was stirred at 20 ℃ for 16 hours. After completion of the reaction, the reaction mixture was poured into water (100mL), stirred for 30min, and extracted with ethyl acetate (100 mL). The organic phase was washed with saturated sodium chloride solution (100mL × 3), dried over anhydrous sodium sulfate and the solvent was evaporated under reduced pressure. The product was purified by silica gel column chromatography (petroleum ether: ethyl acetate 20:1 by volume) to give 6.5g of a colorless oil, which was used directly in the next step.

b) 4-chloro-8- (2, 4-dimethoxybenzyl) -5, 8-dihydropteridin-7 (6H) -one

To a solution of ethyl 2- ((4, 6-dichloropyrimidin-5-yl) amino) acetate (5g) in isopropanol (150mL) at 20 ℃ was added 2, 4-dimethoxybenzylamine (3.67g), followed by triethylamine (4.45 g). The reaction mixture was stirred at 80 ℃ for 18 hours. After the reaction was completed, the reaction solution was filtered, and the filter cake was washed with ethanol and dried under reduced pressure to obtain 5.0g of a near-white solid, which was used directly in the next step.

c) 4-chloro-8- (2, 4-dimethoxybenzyl) -5-methyl-5, 8-dihydropteridin-7 (6H) -one

Methyl iodide (1.68g) was added to a solution of 4-chloro-8- (2, 4-dimethoxybenzyl) -5, 6-dihydropteridin-7 (8H) -one (3.3g) in N, N-dimethylformamide (30mL) at 0 ℃ and the reaction mixture was stirred for 20 min. The temperature was maintained at 0 ℃ and sodium hydride (0.47g) was added. The reaction mixture was stirred for a further 3 hours at 0 ℃. After completion of the reaction, the reaction mixture was poured into water (100mL), and ethyl acetate (100mL) was added thereto for extraction. The organic phase was washed with saturated sodium chloride solution (100mL × 3), dried over anhydrous sodium sulfate, and the solvent was evaporated under reduced pressure. The product was purified by silica gel column chromatography (petroleum ether: ethyl acetate 5:1 by volume) to give 1.5g of the product as a white solid, which was used directly in the next step.

d) 4-chloro-5-methyl-5, 8-dihydropteridin-7 (6H) -one

Trifluoroacetic acid (20mL) was added to 4-chloro-8- (2, 4-dimethoxybenzyl) -5-methyl-5, 6-dihydropteridin-7 (8H) -one (5g) at 20 ℃. The reaction mixture was stirred at 60 ℃ for 16 hours. After completion of the reaction, the solvent was distilled off under reduced pressure to give a near-violet solid (1.0 g). LC-MS (ESI) M/z 199.1(M + H).¹H NMR(300MHz,DMSO-d₆)δ(ppm)11.60(s,1H),8.35(s,1H),3.77(s,2H),2.85(s,3H).

Preparation example 10

Synthesis of 2- (2-amino-3- (3, 4-difluorophenyl) propyl) isoindoline-1, 3-dione

The title compound is prepared starting from tert-butyl (1- (3, 4-difluorophenyl) -3-hydroxypropan-2-yl) carbamate, as described in WO2010093885A1 preparation example 1.

Preparation example 11

Synthesis of 5-chloro-4-methyl-1, 4-dihydro-2H-pyrimido [4,5-d ] [1,3] oxazin-2-one

a)1- (4-amino-6-chloropyrimidin-5-yl) ethanones

1- (4, 6-dichloropyrimidin-5-yl) ethanone (2.5g) was dissolved in tetrahydrofuran (15mL) at 20 ℃, then ammonia (9g) was added, the reaction was stirred at 20 ℃ for 5 hours, then concentrated and diluted with a small amount of water, filtered to give a white solid, and dried under vacuum to give 2g of the white solid, which was used directly in the next step.

b)1- (4-amino-6-chloropyrimidin-5-yl) ethane-1-alcohol is dissolved in methanol (15mL) at the temperature of 20 ℃, 1- (4-amino-6-chloropyrimidin-5-yl) ethanone (1.5g) is cooled to-10 ℃, sodium borohydride (1g) is added in batches, and after the reaction is finished, the temperature of the reaction liquid is slowly raised to 20 ℃ and stirring is continued for 3 hours. After completion of the reaction, the reaction solution was quenched with a saturated aqueous ammonium chloride solution. The reaction was then concentrated and slurried with ethyl acetate (20mL x 2). The mother liquor is concentrated to obtain oily crude product. The crude product was separated by column chromatography to give 400mg of a white oily product. LC-MS (ESI +):174.1(M + H).

c) 5-chloro-4-methyl-1, 4-dihydro-2H-pyrimidine [4,5-d ] [1,3] oxazin-2-one

1- (4-amino-6-chloropyrimidin-5-yl) ethan-1-ol (300mg), N-diisopropylethylamine (282mg) was dissolved in tetrahydrofuran (3mL) at 20 ℃ and then cooled to-5 ℃ and bis (trichloromethyl) carbonate (300mg) was slowly added, and stirred at-5 ℃ for 0.5 hour. Then slowly warmed to 18 ℃ and stirred for 1.5 hours. After completion of the reaction, the reaction was quenched with aqueous sodium bicarbonate and extracted with ethyl acetate (10mL × 3), the organic phases were combined and dried over anhydrous sodium sulfate, filtered and concentrated to give crude oil. The crude product was isolated and purified by column chromatography to give 108mg of a white solid). LC/MS (ESI +): 200.1(M + H).

Preparation example 12

Synthesis of 2- (4-chloro-7-oxo-7, 8-dihydropteridin-5 (6H) -yl) acetonitrile

a)2- (4-chloro-8- (3, 4-dimethoxybenzyl) -7-oxo-7, 8-dihydropteridin-5 (6H) -yl) acetonitrile

To a solution of 4-chloro-8- (2, 4-dimethoxybenzyl) -5, 8-dihydropteridin-7 (6H) -one (2g) in N, N-dimethylformamide (20mL) at 0 deg.C was added sodium hydride (60%, 0.48g), after no bubbles were released bromoacetonitrile (1g) was slowly added dropwise, and the reaction mixture was stirred at 0 deg.C for 3 hours. After completion of the reaction, the reaction mixture was poured into water (100mL), and ethyl acetate (100mL) was added thereto for extraction. The organic phase was washed with saturated sodium chloride solution (100mL × 3), dried over anhydrous sodium sulfate, and the solvent was evaporated under reduced pressure. The product was purified by silica gel column chromatography to give 0.6g of a yellow solid which was used directly in the next step. LC/MS (ESI +):342.1(M + H)

b)2- (4-chloro-7-oxo-7, 8-dihydropteridin-5 (6H) -yl) acetonitrile

2- (4-chloro-8- (3, 4-dimethoxybenzyl) -7-oxo-7, 8-dihydropteridin-5 (6H) -yl) acetonitrile (0.6g) was dissolved in trifluoroacetic acid (15ml) and reacted at 55 ℃ for 4 hours. After the reaction was completed, the pH was adjusted to 8 with a saturated sodium bicarbonate solution, extracted with ethyl acetate (15ml × 4), the organic layers were combined, dried over anhydrous sodium sulfate, the solvent was removed by rotary evaporation under reduced pressure, and purified by column chromatography to obtain a white solid (0.3 g). LC/MS (ESI +):224.1(M + H).

Preparation example 13

Synthesis of 4-chloro-5- (2-methoxyethyl) -5, 8-dihydropteridine-7 (6H) -ketone

a) 4-chloro-8- (2, 4-dimethoxybenzyl) -5- (2-methoxyethyl) -5, 8-dihydropteridin-7 (6H) -one

To a solution of 4-chloro-8- (2, 4-dimethoxybenzyl) -5, 8-dihydropteridin-7 (6H) -one (1g) in N, N-dimethylformamide (10mL) at 0 deg.C was added sodium hydride (60%, 0.24g), 1-iodo-2-methoxyethane (0.8g) was slowly added dropwise after no bubble was released, and the reaction mixture was stirred at 0 deg.C for 3 hours. After completion of the reaction, the reaction mixture was poured into water (100mL), and ethyl acetate (100mL) was added thereto for extraction. The organic phase was washed with saturated sodium chloride solution (100mL × 3), dried over anhydrous sodium sulfate, and the solvent was evaporated under reduced pressure. The product was purified by silica gel column chromatography to give 1.2g of a yellow solid which was used directly in the next step. LC/MS (ESI +):361.2(M + H).

b) 4-chloro-5- (2-methoxyethyl) -5, 8-dihydropteridin-7 (6H) -one

4-chloro-8- (2, 4-dimethoxybenzyl) -5- (2-methoxyethyl) -5, 8-dihydropteridin-7 (6H) -one (1g) was dissolved in trifluoroacetic acid (15ml) and reacted at 55 ℃ for 4 hours. After the reaction was completed, the pH was adjusted to 8 with a saturated sodium bicarbonate solution, extracted with ethyl acetate (15ml × 4), the organic layers were combined, dried over anhydrous sodium sulfate, the solvent was removed by rotary evaporation under reduced pressure, and purified by column chromatography to obtain a white solid (0.5 g).

LC/MS(ESI+):243.2(M+H)。

Example 1(S) -2- (4-chlorophenyl) -3- (isopropylamino) -N- (3- (5-methyl-7-oxo-5, 6,7, 8-tetrahydropteridin-4-yl) phenyl) propionamide

a) Tert-butyl (3- (5-methyl-7-oxo-5, 6,7, 8-tetrahydropteridin-4-yl) phenyl) carbamate

Dissolving 300mg of 4-chloro-5-methyl-5, 8-dihydropteridine-7 (6H) -ketone, 540mg of (3- ((tert-butoxycarbonyl) amino) phenyl) boric acid, 105mg of [1,1' -bis (diphenylphosphino) ferrocene ] palladium dichloride and 980mg of cesium carbonate in a mixed solution of 20mL of dioxane and 2mL of water, heating to reflux under the protection of nitrogen, stopping heating after 5H, and performing column chromatography after decompression drying of a reaction solution to obtain 210mg of light yellow solid;

b)4- (3-aminophenyl) -5-methyl-5, 8-dihydropteridin-7 (6H) -one

Dissolving the tert-butyl (3- (5-methyl-7-oxo-5, 6,7, 8-tetrahydropteridine-4-yl) phenyl) carbamate prepared in the last step in 10mL of trifluoroacetic acid, refluxing for 1h, then evaporating to dryness under reduced pressure, neutralizing with 10mL of saturated sodium bicarbonate solution, and evaporating to dryness and carrying out column chromatography to obtain 110mg of yellow solid;

c) tert-butyl (S) - (2- (4-chlorophenyl) -3- ((3- (5-methyl-7-oxo-5, 6,7, 8-tetrahydropteridin-4-yl) phenyl) amino) -3-oxopropyl) (isopropyl) carbamate

Dissolving 110mg of 4- (3-aminophenyl) -5-methyl-5, 8-dihydropteridine-7 (6H) -one trifluoroacetate, 150mg of (S) -3- ((tert-butoxycarbonyl) (isopropyl) amino) -2- (4-chlorophenyl) propionic acid and 200mg of N, N-diisopropylethylamine in 10mL of dichloromethane, adding 400mg of benzotriazol-1-yloxytris (dimethylamino) phosphonium hexafluorophosphate at 25 ℃, reacting for 1H with stirring, evaporating to dryness under reduced pressure, and performing column chromatography to obtain 197mg of pale yellow solid;

d) (S) -2- (4-chlorophenyl) -3- (isopropylamino) -N- (3- (5-methyl-7-oxo-5, 6,7, 8-tetrahydropteridin-4-yl) phenyl) propanamide

Tert-butyl (S) - (2- (4-chlorophenyl) -3- ((3- (5-methyl-7-oxo-5, 6,7, 8-tetrahydropteridin-4-yl) phenyl) amino) -3-oxopropyl) (isopropyl) carbamate (197mg) was dissolved in trifluoroacetic acid (10ml) and evaporated to dryness under reduced pressure after refluxing for 1h to give a yellow solid (50 mg).

LC/MS(ESI+):479.2(M+H).

¹H NMR(400MHz,CDCl₃)δ10.90(s,1H),8.59(d,J＝1.9Hz,1H),8.16(s,1H),7.72(s,1H),7.56(s,1H),7.46–7.31(m,4H),4.25(s,1H),3.93(d,J＝17.2Hz,1H),3.83(d,J＝16.9Hz,1H),3.72(qd,J＝7.1,1.1Hz,1H),3.49(d,J＝1.6Hz,2H),3.16–2.93(m,2H),2.47(s,3H),1.23(ddd,J＝16.4,8.6,6.1Hz,7H).

Example 2(S) -2- (4-chlorophenyl) -3- (isopropylamino) -N- (4- (5-methyl-7-oxo-5, 6,7, 8-tetrahydropteridin-4-yl) phenyl) propionamide

a) Tert-butyl (4- (5-methyl-7-oxo-5, 6,7, 8-tetrahydropteridin-4-yl) phenyl) carbamate

With reference to the procedure of step a of example 1, the synthesis was carried out starting from 4-chloro-5-methyl-5, 8-dihydropteridin-7 (6H) -one and (4- ((tert-butoxycarbonyl) amino) phenyl) boronic acid.

b)4- (4-aminophenyl) -5-methyl-5, 8-dihydropteridin-7 (6H) -one

With reference to the procedure of step b) of example 1, synthesis from tert-butyl (4- (5-methyl-7-oxo-5, 6,7, 8-tetrahydropteridin-4-yl) phenyl) carbamate.

c) Tert-butyl (S) - (2- (4-chlorophenyl) -3- ((4- (5-methyl-7-oxo-5, 6,7, 8-tetrahydropteridin-4-yl) phenyl) amino) -3-oxopropyl) (isopropyl) carbamate

With reference to the procedure of example 1, step c), was synthesized from 4- (4-aminophenyl) -5-methyl-5, 8-dihydropteridin-7 (6H) -one trifluoroacetate salt and (S) -3- ((tert-butoxycarbonyl) (isopropyl) amino) -2- (4-chlorophenyl) propionic acid.

d) (S) -2- (4-chlorophenyl) -3- (isopropylamino) -N- (4- (5-methyl-7-oxo-5, 6,7, 8-tetrahydropteridin-4-yl) phenyl) propanamide

With reference to the procedure of example 1, step d), was synthesized from tert-butyl (S) - (2- (4-chlorophenyl) -3- ((4- (5-methyl-7-oxo-5, 6,7, 8-tetrahydropteridin-4-yl) phenyl) amino) -3-oxopropyl) (isopropyl) carbamate.

LC/MS(ESI+):479.2(M+H).

¹H NMR(400MHz,DMSO)δ10.60(s,1H),8.51(s,1H),8.32(s,1H),8.06(d,J＝8.5Hz,2H),7.72(d,J＝8.5Hz,2H),7.42(s,4H),3.94(s,1H),3.81(s,2H),3.33-3.28(m,1H),2.87-2.83(m,2H),2.42(s,3H),1.03(s,3H),1.02(s,3H).

Example 3(S) -2- (4-chlorophenyl) -3- (isopropylamino) -N- (4- ((R) -5-methyl-7-oxo-5, 6,7, 8-tetrahydropyrido [2,3-d ] pyrimidin-4-yl) phenyl) propanamide

a) (R) - (4- (5-methyl-7-oxo-5, 6,7, 8-tetrahydropyrido [2,3-d ] pyrimidin-4-yl) phenyl) carbamic acid tert-butyl ester

With reference to the procedure of step a of example 1, synthesis was performed starting from (R) -4-chloro-5-methyl-5, 8-dihydropyrido [2,3-d ] pyrimidin-7 (6H) -one and (4- ((tert-butoxycarbonyl) amino) phenyl) boronic acid.

b) (R) -4- (4-aminophenyl) -5-methyl-5, 8-dihydropyrido [2,3-d ] pyrimidin-7 (6H) -one

Prepared from the product of step a, according to the procedure of step b of example 1.

c) Tert-butyl ((S) -2- (4-chlorophenyl) -3- ((4- ((R) -5-methyl-7-oxo-5, 6,7, 8-tetrahydropyrido [2,3-d ] ] pyrimidin-4-yl) phenyl) amino) -3-oxopropyl) (isopropyl) carbamate

Synthesized according to the procedure of step c of example 1.

d) (S) -2- (4-chlorophenyl) -3- (isopropylamino) -N- (4- ((R) -5-methyl-7-oxo-5, 6,7, 8-tetrahydropyrido [2,3-d ] pyrimidin-4-yl) phenyl) propanamide

Prepared from the product of step c, according to the method of step d of example 1.

LC/MS(ESI+):478.2(M+H).

¹H NMR(400MHz,DMSO)δ11.17(s,1H),10.88(s,1H),9.18(s,1H),8.75(s,1H),8.58(s,1H),7.81(d,J＝8.6Hz,2H),7.60–7.43(m,6H),4.41(d,J＝4.6Hz,1H),3.71(m,1H),3.39(s,1H),3.38–3.24(m,2H),3.14–3.02(m,1H),2.95(dd,J＝16.3,6.0Hz,1H),2.32(d,J＝15.7Hz,1H),1.27(s,3H),1.26(s,3H),0.94(d,J＝7.0Hz,3H).

Example 4(S) -N- (1-amino-3- (3-fluorophenyl) propan-2-yl) -5- (5-methyl-7-oxo-5, 6,7, 8-tetrahydropyridin-4-yl) thiophene-3-carboxamide

a)5- (5-methyl-7-oxo-5, 6,7, 8-tetrahydropteridin-4-yl) thiophene-3-carboxylic acid methyl ester

Methyl 5- (4,4,5, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) thiophene-3-carboxylate (485mg), 4-chloro-5-methyl-5, 8-dihydropteridin-7 (6H) -one (180mg), bis (triphenylphosphine) palladium dichloride (63mg) and cesium carbonate (586mg) were dissolved in a mixed solution of dioxane (20mL) and water (2mL), heated to reflux under nitrogen protection, heating was stopped after 8H, the reaction solution was evaporated to dryness under reduced pressure, and column chromatography was performed to obtain a pale yellow solid (240 mg).

LC/MS(ESI+):305.1(M+H).

¹H NMR(400MHz,CDCl₃)δ9.03(s,1H),8.64(s,1H),8.60(d,J＝1.3Hz,1H),8.29(d,J＝1.4Hz,1H),3.92(s,3H),3.82(m,2H),2.77(s,3H).

b)5- (5-methyl-7-oxo-5, 6,7, 8-tetrahydropteridin-4-yl) thiophene-3-carboxylic acid

Dissolving 150mg of 5- (5-methyl-7-oxo-5, 6,7, 8-tetrahydropteridine-4-yl) thiophene-3-carboxylic acid methyl ester in 10mL of methanol, adding 10mL of 2M lithium hydroxide solution, stirring at room temperature overnight, removing the methanol by rotary evaporation, adding 2M hydrochloric acid to adjust the pH value to 4-5, extracting with ethyl acetate, taking an organic phase, drying with anhydrous sodium sulfate, and performing column chromatography to obtain 55mg of a light yellow solid.

c) (S) -N- (1- (1, 3-dioxoisoindolin-2-yl) -3- (3-fluorophenyl) propan-2-yl) -5- (5-methyl-7-oxo-5, 6,7, 8-tetrahydropteridin-4-yl) thiophene-3-carboxamide

50mg of 5- (5-methyl-7-oxo-5, 6,7, 8-tetrahydropteridin-4-yl) thiophene-3-carboxylic acid, 70mg of (S) -2- (2-amino-3- (3-fluorophenyl) propyl) isoindoline-1, 3-dione, 113mg of benzotriazol-1-yloxytris (dimethylamino) phosphonium hexafluorophosphate and 52mg of N, N-diisopropylethylamine were dissolved in 5mL of N, N-dimethylformamide and stirred at room temperature overnight. Pouring the reaction solution into water, separating out solid, filtering, drying, and purifying by column chromatography to obtain 52mg of light yellow solid.

d) (S) -N- (1-amino-3- (3-fluorophenyl) propan-2-yl) -5- (5-methyl-7-oxo-5, 6,7, 8-tetrahydrodishridin-4-yl) thiophene-3-carboxamide

Dissolving 50mg of the product obtained in the step c in 5mL of methanol, adding 44mg of hydrazine hydrate, stirring overnight at room temperature, evaporating the reaction solution under reduced pressure, and performing column chromatography to obtain 14mg of off-white solid.

LC/MS(ESI+):441.1(M+H).

¹H NMR(400MHz,DMSO-d₆)δ8.57(s,1H),8.44(d,J＝1.5Hz,1H),8.33(d,J＝1.3Hz,1H),8.30(s,1H),7.36–7.23(m,2H),7.10(d,J＝8.7Hz,2H),4.19(s,2H),3.77(m,2H),2.97–2.92(m,1H),2.79(m,2H),2.61(s,3H).

Example 5N- ((S) -1-amino-3- (3-fluorophenyl) propan-2-yl) -4- ((R) -5-methyl-7-oxo-5, 6,7, 8-tetrahydropyrido [2,3-d ] pyrimidin-4-yl) thiophene-2-carboxamide

a) (R) -4- (5-methyl-7-oxo-5, 6,7, 8-tetrahydropyrido [2,3-d ] pyrimidin-4-yl) thiophene-2-carboxylic acid methyl ester

With reference to the procedure of example 4, step a, starting from methyl 4- (4,4,5, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) thiophene-2-carboxylate and (R) -4-chloro-5-methyl-5, 8-dihydropyrido [2,3-d ] pyrimidin-7 (6H) -one.

b) (R) -4- (5-methyl-7-oxo-5, 6,7, 8-tetrahydropyrido [2,3-d ] pyrimidin-4-yl) thiophene-2-carboxylic acid

Prepared from the product of step a, according to the procedure of step b of example 4.

c) Synthesis of (S) -N- (1- (1, 3-dioxoisoindolin-2-yl) -3- (3-fluorophenyl) propan-2-yl) -4- ((R) -5-methyl-7-oxo-5, 6,7, 8-tetrahydropyrido [2,3-d ] pyrimidin-4-yl) thiophene-2-carboxamide

Prepared by reacting the product of step b with (S) -2- (2-amino-3- (3-fluorophenyl) propyl) isoindoline-1, 3-dione, according to the method of step c of example 4. LC/MS (ESI +):570.2(M + H)

d) Synthesis of N- ((S) -1-amino-3- (3-fluorophenyl) propan-2-yl) -4- ((R) -5-methyl-7-oxo-5, 6,7, 8-tetrahydropyrido [2,3-d ] pyrimidin-4-yl) thiophene-2-carboxamide

Prepared from the product of step c, according to the method of step d of example 4.

LC/MS(ESI+):440.2(M+H)

¹H NMR(400MHz,DMSO-d₆)δ8.72(s,1H),8.43(d,J＝8.8Hz,1H),8.19(d,J＝1.3Hz,1H),8.15(d,J＝1.3Hz,1H),7.33–7.25(m,1H),7.12–7.03(m,2H),6.99(td,J＝8.3,1.8Hz,1H),4.06(d,J＝5.5Hz,1H),3.51(d,J＝5.8Hz,1H),2.93(m,3H),2.79(m,1H),2.68(d,J＝6.1Hz,2H),2.39(d,J＝1.7Hz,1H),2.35(d,J＝1.6Hz,1H),1.13(d,J＝7.1Hz,3H).

Example 6(S) -N- (1-amino-3- (3-fluorophenyl) propan-2-yl) -4- (5-methyl-7-oxo-5, 6,7, 8-tetrahydropyridin-4-yl) furan-2-carboxamide

With reference to the procedure of example 4, starting from 4-chloro-5-methyl-5, 8-dihydropteridin-7 (6H) -one and methyl 4- (4,4,5, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) furan-2-carboxylate.

LC/MS(ESI+):425.2(M+H).

¹H NMR(400MHz,DMSO-d₆)δ8.66(s,2H),8.50(d,J＝8.6Hz,1H),7.90(s,1H),7.36(td,J＝7.9,6.1Hz,1H),7.21–7.13(m,2H),7.05(td,J＝8.9,8.4,2.1Hz,1H),4.18(d,J＝6.6Hz,2H),3.79(s,3H),3.01(dd,J＝13.7,5.3Hz,1H),2.95–2.70(m,3H),2.65(s,3H).

Example 7(S) -N- (1-amino-3- (3-fluorophenyl) propan-2-yl) -4- (5-methyl-7-oxo-5, 6,7, 8-tetrahydropyridin-4-yl) thiophene-2-carboxamide

a) Synthesis of methyl 4- (5-methyl-7-oxo-5, 6,7, 8-tetrahydropteridin-4-yl) thiophene-2-carboxylate

With reference to the procedure of example 4, step a, starting from methyl 4- (4,4,5, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) thiophene-2-carboxylate and 4-chloro-5-methyl-5, 8-dihydropteridin-7 (6H) -one.

b) Synthesis of 4- (5-methyl-7-oxo-5, 6,7, 8-tetrahydropteridin-4-yl) thiophene-2-carboxylic acid

c) Synthesis of (S) -N- (1- (1, 3-dioxoisoindolin-2-yl) -3- (3-fluorophenyl) propan-2-yl) -4- (5-methyl-7-oxo-5, 6,7, 8-tetrahydropteridin-4-yl) thiophene-2-carboxamide

Prepared by reacting the product of step b with (S) -2- (2-amino-3- (3-fluorophenyl) propyl) isoindoline-1, 3-dione, according to the method of step c of example 4.

d) Synthesis of (S) -N- (1-amino-3- (3-fluorophenyl) propan-2-yl) -4- (5-methyl-7-oxo-5, 6,7, 8-tetrahydronaphthyridin-4-yl) thiophene-2-carboxamide

LC/MS(ESI+):441.1(M+H).

¹H NMR(400MHz,DMSO-d₆)δ8.76(m,1H),8.66(m,1H),8.60(m,1H),8.55(m,1H),7.30(m,1H),7.10(m,2H),7.05(m,1H),4.33(m,2H),3.77(s,2H),2.98–2.84(m,4H),2.68(s,3H).

Example 8(S) -N- (1-amino-3- (3-fluorophenyl) propan-2-yl) -5-methyl-4- (5-methyl-7-oxo-5, 6,7, 8-tetrahydrodishridin-4-yl) thiophene-2-carboxamide

a) Synthesis of methyl 5-methyl-4- (5-methyl-7-oxo-5, 6,7, 8-tetrahydropteridin-4-yl) thiophene-2-carboxylate

Referring to the procedure of step a of example 4, prepared from methyl 5-methyl-4- (4,4,5, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) thiophene-2-carboxylate synthesized in preparation 4 by reaction with 4-chloro-5-methyl-5, 8-dihydropteridin-7 (6H) -one.

b) Synthesis of 5-methyl-4- (5-methyl-7-oxo-5, 6,7, 8-tetrahydropteridin-4-yl) thiophene-2-carboxylic acid

c) Synthesis of (S) -N- (1- (1, 3-dioxoisoindolin-2-yl) -3- (3-fluorophenyl) propan-2-yl) -5-methyl-4- (5-methyl-7-oxo-5, 6,7, 8-tetrahydropteridin-4-yl) thiophene-2-carboxamide

d) (S) -N- (1-amino-3- (3-fluorophenyl) propan-2-yl) -5-methyl-4- (5-methyl-7-oxo-5, 6,7, 8-tetrahydronaphthyridin-4-yl) thiophene-2-carboxamide

LC/MS(ESI+):455.1(M+H).

¹H NMR(400MHz,DMSO-d₆)δ8.43(s,1H),8.30(d,J＝8.4Hz,1H),7.86(s,1H),7.30(td,J＝8.0,6.2Hz,1H),7.13–7.03(m,2H),7.00(td,J＝8.6,2.5Hz,1H),4.13(t,J＝6.9Hz,1H),3.88(d,J＝1.4Hz,2H),2.93(dd,J＝13.6,5.4Hz,1H),2.86–2.66(m,3H),2.40(s,3H),2.36(s,3H),1.32–1.21(m,2H).

Example 9(S) -N- (1-amino-3- (3-fluorophenyl) propan-2-yl) -3-methyl-4- (5-methyl-7-oxo-5, 6,7, 8-tetrahydrodishridin-4-yl) thiophene-2-carboxamide

Detailed description of the inventionthe preparation of the compound of formula i according to example 4 is carried out starting from 4-chloro-5-methyl-5, 8-dihydropteridin-7 (6H) -one and 3-methyl-4- (4,4,5, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) thiophene-2-carboxylate.

LC/MS(ESI+):455.1(M+H)..

¹H NMR(400MHz,Chloroform-d)δ8.52(s,1H),7.57(s,1H),7.11–6.90(m,3H),6.52(d,J＝8.0Hz,1H),4.34(d,J＝7.0Hz,1H),3.87(s,2H),3.03(dd,J＝13.8,6.5Hz,1H),2.97–2.79(m,3H),2.51(s,3H),2.38(s,3H).

Example 10(S) -N- (1-amino-3- (3-fluorophenyl) propan-2-yl) -3-fluoro-4- (5-methyl-7-oxo-5, 6,7, 8-tetrahydrodishridin-4-yl) thiophene-2-carboxamide

Detailed description of the inventionthe preparation of the compound of formula i according to example 4 is carried out starting from 4-chloro-5-methyl-5, 8-dihydropteridin-7 (6H) -one and 3-fluoro-4- (4,4,5, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) thiophene-2-carboxylic acid methyl ester.

LC/MS(ESI+):459.2(M+H).

¹H NMR(400MHz,DMSO-d₆)δ8.43(s,1H),8.36(s,1H),8.13(d,J＝4.0Hz,1H),7.88(s,2H),7.32(q,J＝7.5Hz,1H),7.15–6.94(m,3H),4.19(s,1H),3.86(s,2H),3.00–2.65(m,5H),2.51(s,3H).

Example 11(S) -N- (1-amino-3- (3-fluorophenyl) propan-2-yl) -5- (5-methyl-7-oxo-5, 6,7, 8-tetrahydropyridin-4-yl) thiophene-2-carboxamide

a) Synthesis of methyl 5- (5-methyl-7-oxo-5, 6,7, 8-tetrahydropteridin-4-yl) thiophene-2-carboxylate

With reference to the procedure of example 4, step a, prepared from methyl 5- (4,4,5, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) thiophene-2-carboxylate by reaction with 4-chloro-5-methyl-5, 8-dihydropteridin-7 (6H) -one.

b) Synthesis of 5- (5-methyl-7-oxo-5, 6,7, 8-tetrahydropteridin-4-yl) thiophene-2-carboxylic acid

c) Synthesis of (S) -N- (1- (1, 3-dioxoisoindolin-2-yl) -3- (3-fluorophenyl) propan-2-yl) -5- (5-methyl-7-oxo-5, 6,7, 8-tetrahydropteridin-4-yl) thiophene-2-carboxamide

Prepared by reacting the product of step b with (S) -2- (2-amino-3- (3-fluorophenyl) propyl) isoindoline-1, 3-dione, according to the method of step c of example 4. LC/MS (ESI +):571.2(M + H).

d) Synthesis of (S) -N- (1-amino-3- (3-fluorophenyl) propan-2-yl) -5- (5-methyl-7-oxo-5, 6,7, 8-tetrahydronaphthyridin-4-yl) thiophene-2-carboxamide

LC/MS(ESI+):441.2(M+H).

¹H NMR(400MHz,DMSO-d₆)δ8.57(s,1H),8.49(s,1H),8.36(s,1H),8.13(d,J＝3.9Hz,1H),7.83(d,J＝4.1Hz,1H),7.31(q,J＝7.4Hz,1H),7.10(d,J＝8.1Hz,2H),7.05–6.95(m,1H),4.13(s,1H),3.77(s,2H),3.00–2.92(m,1H),2.79(dd,J＝35.3,7.7Hz,3H),2.62(s,3H).

EXAMPLE 12 Synthesis of (S) -N- (1-amino-3- (3-fluorophenyl) propan-2-yl) -4-methyl-5- (5-methyl-7-oxo-5, 6,7, 8-tetrahydrodishridin-4-yl) thiophene-3-carboxamide

Specific preparation method referring to example 4, prepared from 4-methyl-5- (4,4,5, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) thiophene-3-carboxylic acid methyl ester and 4-chloro-5-methyl-5, 8-dihydropteridin-7 (6H) -one as starting materials.

LC/MS(ESI+):455.1(M+H).

¹H NMR(400MHz,DMSO-d₆)δ8.40(d,J＝16.0Hz,3H),8.03(s,1H),7.33(td,J＝8.0,6.2Hz,1H),7.20–6.93(m,3H),4.26(d,J＝9.4Hz,1H),3.87(s,2H),3.01–2.71(m,4H),2.42(s,3H),2.12(s,3H).

Example 13(S) -N- (1-amino-3- (3-fluorophenyl) propan-2-yl) -3-methyl-5- (5-methyl-7-oxo-5, 6,7, 8-tetrahydrodishridin-4-yl) thiophene-2-carboxamide

a) Synthesis of methyl 3-methyl-5- (5-methyl-7-oxo-5, 6,7, 8-tetrahydropteridin-4-yl) thiophene-2-carboxylate

With reference to the procedure of example 4, step a, starting from methyl 3-methyl-5- (4,4,5, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) thiophene-2-carboxylate and 4-chloro-5-methyl-5, 8-dihydropteridin-7 (6H) -one.

b) Synthesis of 3-methyl-5- (5-methyl-7-oxo-5, 6,7, 8-tetrahydropteridin-4-yl) thiophene-2-carboxylic acid

c) Synthesis of (S) -N- (1- (1, 3-dioxoisoindolin-2-yl) -3- (3-fluorophenyl) propan-2-yl) -3-methyl-5- (5-methyl-7-oxo-5, 6,7, 8-tetrahydropteridin-4-yl) thiophene-2-carboxamide

Prepared by reacting the product of step b with (S) -2- (2-amino-3- (3-fluorophenyl) propyl) isoindoline-1, 3-dione, according to the method of step c of example 4. LC/MS (ESI +):585.2(M + H).

d) Synthesis of (S) -N- (1-amino-3- (3-fluorophenyl) propan-2-yl) -3-methyl-5- (5-methyl-7-oxo-5, 6,7, 8-tetrahydronaphthyridin-4-yl) thiophene-2-carboxamide

LC/MS(ESI+):455.1(M+H).

¹H NMR(400MHz,CDCl₃)δ8.43(s,1H),7.82(s,1H),7.42–7.29(m,1H),7.12–6.93(m,3H),6.66(d,J＝8.1Hz,1H),4.40(q,J＝6.7Hz,1H),3.89–3.57(m,2H),3.14–2.81(m,4H),2.63(s,3H),2.43(s,3H).

Example 14(S) -N- (1-amino-3- (3-fluorophenyl) propan-2-yl) -4-methyl-5- (5-methyl-7-oxo-5, 6,7, 8-tetrahydrodishridin-4-yl) thiophene-2-carboxamide

a) Synthesis of methyl 4-methyl-5- (5-methyl-7-oxo-5, 6,7, 8-tetrahydropteridin-4-yl) thiophene-2-carboxylate

With reference to the procedure of example 4, step a, starting from methyl 4-methyl-5- (4,4,5, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) thiophene-2-carboxylate and 4-chloro-5-methyl-5, 8-dihydropteridin-7 (6H) -one.

b) Synthesis of 4-methyl-5- (5-methyl-7-oxo-5, 6,7, 8-tetrahydropteridin-4-yl) thiophene-2-carboxylic acid

c) Synthesis of (S) -N- (1- (1, 3-dioxoisoindolin-2-yl) -3- (3-fluorophenyl) propan-2-yl) -4-methyl-5- (5-methyl-7-oxo-5, 6,7, 8-tetrahydropteridin-4-yl) thiophene-2-carboxamide

d) (S) -N- (1-amino-3- (3-fluorophenyl) propan-2-yl) -4-methyl-5- (5-methyl-7-oxo-5, 6,7, 8-tetrahydronaphthyridin-4-yl) thiophene-2-carboxamide

LC/MS(ESI+):455.1(M+H)

¹H NMR(400MHz,DMSO-d₆)δ8.62(s,1H),8.47(s,1H),8.39(s,1H),7.65(s,1H),7.35-7.29(m,1H),7.11-7.09(m,2H),7.01(t,J＝8.2Hz,1H),4.25-4.11(m,1H),3.86(s,2H),3.01-2.90(m,2H),2.90-2.74(m,4H),2.47(s,3H),2.28(s,3H).

Example 15(S) -N- (1-amino-3- (3-fluorophenyl) propan-2-yl) -2-methyl-5- (5-methyl-7-oxo-5, 6,7, 8-tetrahydrodishridin-4-yl) thiophene-3-carboxamide

a) Synthesis of methyl 2-methyl-5- (5-methyl-7-oxo-5, 6,7, 8-tetrahydropteridin-4-yl) thiophene-3-carboxylate

With reference to the procedure of example 4, step a, prepared from methyl 2-methyl-5- (4,4,5, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) thiophene-3-carboxylate by reaction with 4-chloro-5-methyl-5, 8-dihydropteridin-7 (6H) -one.

b) 2-methyl-5- (5-methyl-7-oxo-5, 6,7, 8-tetrahydropteridin-4-yl) thiophene-3-carboxylic acid

c) Synthesis of (S) -N- (1- (1, 3-dioxoisoindolin-2-yl) -3- (3-fluorophenyl) propan-2-yl) -2-methyl-5- (5-methyl-7-oxo-5, 6,7, 8-tetrahydropteridin-4-yl) thiophene-3-carboxamide

d) (S) -N- (1-amino-3- (3-fluorophenyl) propan-2-yl) -2-methyl-5- (5-methyl-7-oxo-5, 6,7, 8-tetrahydronaphthyridin-4-yl) thiophene-3-carboxamide

LC/MS(ESI+):445.1(M+H).

¹H NMR(400MHz,DMSO-d₆)δ8.55(s,1H),8.21(s,1H),8.09(t,J＝9.8Hz,1H),7.31(q,J＝7.4Hz,1H),7.13–7.04(m,2H),7.01(t,J＝8.5Hz,1H),6.26(s,2H),4.10(d,J＝7.8Hz,1H),3.74(s,2H),3.17(s,1H),2.96(dd,J＝13.8,5.1Hz,1H),2.84–2.66(m,3H),2.56(s,3H).

Example 16(S) -N- (1-amino-3- (3-fluorophenyl) propan-2-yl) -1-methyl-3- (5-methyl-7-oxo-5, 6,7, 8-tetrahydrodishridin-4-yl) -1H-pyrazole-5-carboxamide

Specific preparation method referring to example 4, starting from methyl 1-methyl-3- (4,4,5, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) -1H-pyrazole-5-carboxylate and 4-chloro-5-methyl-5, 8-dihydropteridin-7 (6H) -one.

LC/MS(ESI+):439.2(M+H)

Example 17(S) -N- (1-amino-3- (3-fluorophenyl) propan-2-yl) -4- (5-methyl-7-oxo-5, 6,7, 8-tetrahydropyridin-4-yl) benzamide

a) Synthesis of methyl 4- (5-methyl-7-oxo-5, 6,7, 8-tetrahydropteridin-4-yl) benzoate

With reference to the procedure of step a of example 4, prepared from 4-chloro-5-methyl-5, 8-dihydropteridin-7 (6H) -one by reaction with (4- (methoxycarbonyl) phenyl) boronic acid.

b) Synthesis of 4- (5-methyl-7-oxo-5, 6,7, 8-tetrahydropteridin-4-yl) benzoic acid

c) Synthesis of (S) -N- (1- (1, 3-dioxoisoindolin-2-yl) -3- (3-fluorophenyl) propan-2-yl) -4- (5-methyl-7-oxo-5, 6,7, 8-tetrahydropteridin-4-yl) benzamide

d) Synthesis of (S) -N- (1-amino-3- (3-fluorophenyl) propan-2-yl) -4- (5-methyl-7-oxo-5, 6,7, 8-tetrahydronaphthyridin-4-yl) benzamide

LC/MS(ESI+):435.2(M+H)

¹H NMR(400MHz,DMSO-d₆)δ8.71(d,J＝8.3Hz,1H),8.53(s,1H),8.39(s,1H),8.01(d,J＝8.2Hz,2H),7.91(d,J＝8.2Hz,2H),7.32(dd,J＝14.4,7.6Hz,1H),7.13(d,J＝7.7Hz,2H),7.01(t,J＝8.4Hz,1H),4.33(d,J＝6.7Hz,1H),3.88(s,2H),3.00–2.84(m,4H),2.40(s,3H).

Example 18(S) -N- (1-amino-3- (3-fluorophenyl) propan-2-yl) -3- (5-methyl-7-oxo-5, 6,7, 8-tetrahydropyridin-4-yl) benzamide

a) Synthesis of methyl 3- (5-methyl-7-oxo-5, 6,7, 8-tetrahydropteridin-4-yl) benzoate

With reference to the procedure of step a of example 4, prepared by reacting 4-chloro-5-methyl-5, 8-dihydropteridin-7 (6H) -one with (3- (methoxycarbonyl) phenyl) boronic acid as starting material.

b) Synthesis of 3- (5-methyl-7-oxo-5, 6,7, 8-tetrahydropteridin-4-yl) benzoic acid

Prepared from the product of step a, according to the procedure of step a of example 4.

c) Synthesis of (S) -N- (1- (1, 3-dioxoisoindolin-2-yl) -3- (3-fluorophenyl) propan-2-yl) -3- (5-methyl-7-oxo-5, 6,7, 8-tetrahydropteridin-4-yl) benzamide

Prepared by reacting the product of step b with (S) -2- (2-amino-3- (3-fluorophenyl) propyl) isoindoline-1, 3-dione, according to the method of step c of example 4. LC/MS (ESI +):565.2(M + H).

d) Synthesis of (S) -N- (1-amino-3- (3-fluorophenyl) propan-2-yl) -3- (5-methyl-7-oxo-5, 6,7, 8-tetrahydronaphthyridin-4-yl) benzamide

LC/MS(ESI+):435.1(M+H).

¹H NMR(400MHz,DMSO-d₆)δ8.55(d,J＝12.3Hz,2H),8.34(d,J＝19.9Hz,2H),8.10(d,J＝7.8Hz,1H),7.88(d,J＝7.8Hz,1H),7.56(t,J＝7.7Hz,1H),7.36–7.26(m,1H),7.14–7.06(m,2H),7.00(td,J＝8.7,2.6Hz,1H),4.27(s,1H),3.94–3.79(m,2H),3.00–2.85(m,1H),2.84(s,2H),2.38(s,3H).

Example 19(S) -N- (1-amino-3- (3-fluorophenyl) propan-2-yl) -3-methyl-5- (5-methyl-7-oxo-5, 6,7, 8-tetrahydrodishridin-4-yl) benzamide

a) Synthesis of methyl 3-methyl-5- (5-methyl-7-oxo-5, 6,7, 8-tetrahydropteridin-4-yl) benzoate

With reference to the procedure of step a of example 4, prepared by reacting 4-chloro-5-methyl-5, 8-dihydropteridin-7 (6H) -one with (3- (methoxycarbonyl) -5-methylphenyl) boronic acid as starting material.

b) Synthesis of 3-methyl-5- (5-methyl-7-oxo-5, 6,7, 8-tetrahydropteridin-4-yl) benzoic acid

c) Synthesis of (S) -N- (1- (1, 3-dioxoisoindolin-2-yl) -3- (3-fluorophenyl) propan-2-yl) -3-methyl-5- (5-methyl-7-oxo-5, 6,7, 8-tetrahydropteridin-4-yl) benzamide

Prepared by reacting the product of step b with (S) -2- (2-amino-3- (3-fluorophenyl) propyl) isoindoline-1, 3-dione, according to the method of step c of example 4. LC/MS (ESI +):579.2(M + H).

d) Synthesis of (S) -N- (1-amino-3- (3-fluorophenyl) propan-2-yl) -3-methyl-5- (5-methyl-7-oxo-5, 6,7, 8-tetrahydronaphthyridin-4-yl) benzamide

¹H NMR(400MHz,DMSO-d₆)δ8.57(d,J＝8.1Hz,1H),8.51(s,1H),8.37(s,1H),8.11(d,J＝1.7Hz,1H),7.87(d,J＝1.7Hz,1H),7.71(d,J＝1.9Hz,1H),7.32(td,J＝8.1,6.3Hz,1H),7.10(dt,J＝8.7,2.3Hz,2H),7.07–6.96(m,1H),4.32(d,J＝7.8Hz,2H),3.98–3.79(m,2H),3.05–2.77(m,3H),2.42(s,3H),2.38(s,3H).LC/MS(ESI+):449.1(M+H).

EXAMPLE 20(S) -N- (1-amino-3- (3-fluorophenyl) propan-2-yl) -2-fluoro-5- (5-methyl-7-oxo-5, 6,7, 8-tetrahydrodishridin-4-yl) benzamide

a) Synthesis of methyl 2-fluoro-5- (5-methyl-7-oxo-5, 6,7, 8-tetrahydropteridin-4-yl) benzoate

With reference to the procedure of step a of example 4, prepared by reacting 4-chloro-5-methyl-5, 8-dihydropteridin-7 (6H) -one with (4-fluoro-3- (methoxycarbonyl) phenyl) boronic acid as starting material.

b) Synthesis of 2-fluoro-5- (5-methyl-7-oxo-5, 6,7, 8-tetrahydropteridin-4-yl) benzoic acid

c) Synthesis of (S) -N- (1- (1, 3-dioxoisoindolin-2-yl) -3- (3-fluorophenyl) propan-2-yl) -2-fluoro-5- (5-methyl-7-oxo-5, 6,7, 8-tetrahydropteridin-4-yl) benzamide

Prepared by reacting the product of step b with (S) -2- (2-amino-3- (3-fluorophenyl) propyl) isoindoline-1, 3-dione, according to the method of step c of example 4. LC/MS (ESI +):583.2(M + H).

d) (S) -N- (1-amino-3- (3-fluorophenyl) propan-2-yl) -2-fluoro-5- (5-methyl-7-oxo-5, 6,7, 8-tetrahydronaphthyridin-4-yl) benzamide

¹H NMR(400MHz,DMSO-d₆)δ9.26(s,1H),8.58(s,1H),8.48(d,J＝8Hz,1H),8.11-7.99(d,J＝8Hz,1H),7.80-7.77(m,1H),7.66-7.57(m,4H),7.24-7.22(d,J＝8Hz,1H),6.90-6.88(d,J＝8Hz,2H),4.15-4.10(m,2H),3.20-3.10(br,3H),2.89-3.05(br,3H).LC/MS(ESI+):453.2(M+H).

Example 21(S) -N- (1-amino-3- (3-fluorophenyl) propan-2-yl) -3-fluoro-5- (5-methyl-7-oxo-5, 6,7, 8-tetrahydrodishridin-4-yl) benzamide

a) Synthesis of methyl 3-fluoro-5- (5-methyl-7-oxo-5, 6,7, 8-tetrahydropteridin-4-yl) benzoate

With reference to the procedure of step a of example 4, prepared by reacting 4-chloro-5-methyl-5, 8-dihydropteridin-7 (6H) -one with (3-fluoro-5- (methoxycarbonyl) phenyl) boronic acid as starting material.

b) Synthesis of 3-fluoro-5- (5-methyl-7-oxo-5, 6,7, 8-tetrahydropteridin-4-yl) benzoic acid

c) Synthesis of (S) -N- (1- (1, 3-dioxoisoindolin-2-yl) -3- (3-fluorophenyl) propan-2-yl) -3-fluoro-5- (5-methyl-7-oxo-5, 6,7, 8-tetrahydropteridin-4-yl) benzamide

d) Synthesis of (S) -N- (1-amino-3- (3-fluorophenyl) propan-2-yl) -3-fluoro-5- (5-methyl-7-oxo-5, 6,7, 8-tetrahydronaphthyridin-4-yl) benzamide

LC/MS(ESI+):453.2(M+H)

¹H NMR(400MHz,DMSO-d₆)δ9.44(s,1H),8.64(s,1H),8.21-8.19(d,J＝8Hz,1H),8.10-7.99(d,J＝8Hz,1H),7.80-7.77(m,1H),7.66-7.57(m,5H),7.24-7.22(d,J＝8Hz,1H),6.90-6.88(d,J＝8Hz,2H),4.15-4.10(m,2H),3.20-3.10(br,3H),2.89-3.05(br,3H).

Example 22(S) -N- (1-amino-3- (3-fluorophenyl) propan-2-yl) -4-fluoro-5- (5-methyl-7-oxo-5, 6,7, 8-tetrahydrodishridin-4-yl) benzamide

The preparation method is characterized by referring to example 4, and the compound is prepared by using 4-chloro-5-methyl-5, 8-dihydropteridine-7 (6H) -ketone and (2-fluoro-5- (methoxycarbonyl) phenyl) boric acid as starting materials.

LC/MS(ESI+):453.2(M+H)

¹H NMR(400MHz,DMSO)δ8.43(s,1H),8.29(d,J＝8.3Hz,1H),8.02(dt,J＝9.2,4.6Hz,1H),7.97–7.89(m,1H),7.41(t,J＝9.1Hz,1H),7.29(dd,J＝14.3,7.8Hz,1H),7.11–7.04(m,2H),6.98(t,J＝8.6Hz,1H),4.10(d,J＝5.8Hz,1H),3.87(d,J＝17.0Hz,2H),2.95(dd,J＝13.6,5.3Hz,1H),2.84–2.74(m,1H),2.67(d,J＝6.0Hz,2H),2.37(s,4H).

Example 23N- ((S) -1-amino-3- (3-fluorophenyl) propan-2-yl) -5- ((R) -5-methyl-7-oxo-5, 6,7, 8-tetrahydropyrido [2,3-d ] pyrimidin-4-yl) nicotinamide

The preparation method is that (R) -4-chloro-5-methyl-5, 8-dihydropyrido [2,3-d ] pyrimidin-7 (6H) -one and (5- (methoxycarbonyl) pyridin-3-yl) boric acid are used as starting materials for reaction, and the preparation method is that according to example 4.

LC/MS(ESI+):435.1(M+H).

¹H NMR(400MHz,DMSO-d₆)δ9.05(s,1H),8.91(s,1H),8.80(s,1H),8.51(d,J＝8.0Hz,1H),8.31(s,1H),7.29(q,J＝7.4Hz,1H),7.16–7.04(m,2H),6.99(t,J＝8.3Hz,1H),4.13(s,1H),3.29–3.09(m,3H),2.98(dq,J＝13.9,4.8Hz,2H),2.80(dd,J＝13.6,8.9Hz,1H),2.70(d,J＝6.1Hz,2H),2.35(d,J＝16.2Hz,1H),1.00(d,J＝7.0Hz,3H).

Example 24N- ((S) -1-amino-3- (3-fluorophenyl) propan-2-yl) -2-fluoro-5- ((R) -5-methyl-7-oxo-5, 6,7, 8-tetrahydropyrido [2,3-d ] pyrimidin-4-yl) benzamide

a) Synthesis of methyl (R) -2-fluoro-5- (5-methyl-7-oxo-5, 6,7, 8-tetrahydropyrido [2,3-d ] pyrimidin-4-yl) benzoate

With reference to the procedure of step a of example 4, synthesis was performed starting from (4-fluoro-3- (methoxycarbonyl) phenyl) boronic acid and (R) -4-chloro-5-methyl-5, 8-dihydropyrido [2,3-d ] pyrimidin-7 (6H) -one.

b) Synthesis of (R) -2-fluoro-5- (5-methyl-7-oxo-5, 6,7, 8-tetrahydropyrido [2,3-d ] pyrimidin-4-yl) benzoic acid

c) Synthesis of N- ((S) -1- (1, 3-dioxoisoindolin-2-yl) -3- (3-fluorophenyl) propan-2-yl) -2-fluoro-5- ((R) -5-methyl-7-oxo-5, 6,7, 8-tetrahydropyrido [2,3-d ] pyrimidin-4-yl) benzamide

d) N- ((S) -1-amino-3- (3-fluorophenyl) propan-2-yl) -2-fluoro-5- ((R) -5-methyl-7-oxo-5, 6,7, 8-tetrahydropyrido [2,3-d ] pyrimidin-4-yl) benzamide

LC/MS(ESI+):452.1(M+H)

¹H NMR(400MHz,DMSO-d₆)δ8.73(s,1H),8.28(d,J＝8.4Hz,1H),7.75-7.68(m,1H),7.66(d,J＝6.4Hz,1H),7.47-7.40(m,1H),7.33-7.26(m,1H),7.12-7.05(m,2H),7.00(t,J＝8.5Hz,1H),4.35-4.17(m,1H),4.10(d,J＝6.0Hz,1H),3.28-3.24(m,1H),3.02-2.96(m,1H),2.93(s,2H),2.78-2.72(m,2H),2.67(d,J＝5.8Hz,2H),2.34(d,J＝15.9Hz,1H),0.97(d,J＝7.0Hz,3H).

Example 25(S) -N- (1-amino-3- (3-fluorophenyl) propan-2-yl) -5- (5-methyl-7-oxo-5, 6,7, 8-tetrahydrodishridin-4-yl) nicotinamide

a) Synthesis of methyl 5- (5-methyl-7-oxo-5, 6,7, 8-tetrahydropteridin-4-yl) nicotinate

With reference to the procedure of step a of example 4, prepared by reacting 4-chloro-5-methyl-5, 8-dihydropteridin-7 (6H) -one with (5- (methoxycarbonyl) pyridin-3-yl) boronic acid as starting material.

b)5- (5-methyl-7-oxo-5, 6,7, 8-tetrahydropteridin-4-yl) nicotinic acid

Prepared by reacting the product of step a according to the procedure of step b of example 4. LC/MS (ESI +) 286.1[ M + H ] +.

c) Synthesis of (S) -N- (1- (1, 3-dioxoisoindolin-2-yl) -3- (3-fluorophenyl) propan-2-yl) -5- (5-methyl-7-oxo-5, 6,7, 8-tetrahydropteridin-4-yl) nicotinamide

Prepared by reacting the product of step b with (S) -2- (2-amino-3- (3-fluorophenyl) propyl) isoindoline-1, 3-dione, according to the method of step c of example 4. LC/MS (ESI +):566.2[ M + H ] +.

d) (S) -N- (1-amino-3- (3-fluorophenyl) propan-2-yl) -5- (5-methyl-7-oxo-5, 6,7, 8-tetrahydrodishridin-4-yl) nicotinamide

LC/MS(ESI+):436.2(M+H).

Example 26(S) -N- (1-amino-3- (3-fluorophenyl) propan-2-yl) -3- (5- (2-methoxyethyl) -7-oxo-5, 6,7, 8-tetrahydrodishrid-4-yl) benzamide

Detailed description of the inventionthe title compound was prepared starting with 4-chloro-5- (2-methoxyethyl) -5, 8-dihydropteridin-7 (6H) -one prepared in preparation 13 and (3- (methoxycarbonyl) phenyl) boronic acid in reference to example 4.

LC/MS(ESI+):479.1(M+H)

Example 27N- ((S) -1-amino-3- (3-fluorophenyl) propan-2-yl) -3- (4-methyl-2-oxo-1, 4-dihydro-2H-pyrimido [4,5-d ] [1,3] oxazin-5-yl) benzamide

Detailed description of the inventionthe title compound was prepared by starting with 5-chloro-4-methyl-1, 4-dihydro-2H-pyrimido [4,5-d ] [1,3] oxazin-2-one prepared in preparation 11 and (3- (methoxycarbonyl) phenyl) boronic acid in accordance with example 4.

LC/MS(ESI+):436.1(M+H).

¹H NMR(400MHz,DMSO-d₆)δ8.85(s,1H),8.31(d,J＝8.4Hz,1H),8.01–7.87(m,2H),7.79–7.68(m,1H),7.61(t,J＝7.7Hz,1H),7.35–7.21(m,1H),7.15–7.03(m,2H),6.97(tt,J＝7.9,3.3Hz,1H),5.74(dq,J＝10.2,6.7Hz,1H),4.12(td,J＝8.1,5.0Hz,1H),3.06–2.74(m,2H),2.68(dd,J＝6.1,2.1Hz,2H),1.33(dd,J＝6.7,4.2Hz,3H).

Example 28(S) -N- (1-amino-3- (3-fluorophenyl) propan-2-yl) -3- (5- (2-hydroxyethyl) -7-oxo-5, 6,7, 8-tetrahydrodishrid-4-yl) benzamide

a)5- (2- ((tert-butyldimethylsilyloxy) ethyl) -4-chloro-8- (2, 4-dimethoxybenzyl) -5, 8-dihydropteridin-7 (6H) -one

To a solution of 4-chloro-8- (2, 4-dimethoxybenzyl) -5, 8-dihydropteridin-7 (6H) -one (1g) in N, N-dimethylformamide (10mL) was added sodium hydride (60%, 0.24g) at 0 deg.C, after no bubble was released, (2-bromoethoxy) (tert-butyl) dimethylsilane (0.7g) was slowly added dropwise, and the reaction mixture was stirred at 0 deg.C for 3 hours. After completion of the reaction, the reaction mixture was poured into water (100mL), and ethyl acetate (100mL) was added thereto for extraction. The organic phase was washed with saturated sodium chloride solution (100mL × 3), dried over anhydrous sodium sulfate, and the solvent was evaporated under reduced pressure. The product was purified by silica gel column chromatography to give 0.3g of a yellow solid which was used directly in the next step.

b)3- (5- (2- ((tert-Butyldimethylsilanyloxy) ethyl) -8- (2, 4-dimethoxybenzyl) -7-oxo-5, 6,7, 8-tetrahydronaphthyridin-4-yl) benzoic acid methyl ester

Dissolving the product (0.3g) obtained in the step a, 3- (methoxycarbonyl) phenyl) boric acid (0.2g), palladium [1,1' -bis (diphenylphosphino) ferrocene ] dichloride (47mg) and cesium carbonate (0.4g) in a mixed solution of dioxane (20mL) and water (2mL), heating to reflux under the protection of nitrogen, stopping heating after 8h, decompressing and evaporating the reaction solution, and performing column chromatography to obtain a light yellow solid (240 mg). .

c)3- (5- (2-hydroxyethyl) -7-oxo-5, 6,7, 8-tetrahydronaphthyridin-4-yl) benzoic acid methyl ester

The product of step b (260mg) was dissolved in trifluoroacetic acid (5ml) and reacted at 55 ℃ for 4 hours. After the reaction was completed, the pH was adjusted to 8 with a saturated sodium bicarbonate solution, extracted with ethyl acetate (5ml × 4), the organic layers were combined, dried over anhydrous sodium sulfate, the solvent was removed by rotary evaporation under reduced pressure, and purified by column chromatography to obtain a white solid product (98 mg).

d)3- (5- (2-hydroxyethyl) -7-oxo-5, 6,7, 8-tetrahydropydrin-4-yl) benzoic acid

Dissolving 98mg of the product obtained in the step c in 10mL of methanol, adding 10mL of 2M lithium hydroxide solution, stirring at room temperature overnight, performing rotary evaporation to remove the methanol, adding 2M hydrochloric acid to adjust the pH value to 4-5, extracting with EA, taking an organic phase, drying with anhydrous sodium sulfate, and performing column chromatography to obtain 55mg of a light yellow solid.

e) (S) -N- (1- (1, 3-dioxoisoindolin-2-yl) -3- (3-fluorophenyl) propan-2-yl) -3- (5- (2-hydroxyethyl) -7-oxo-5, 6,7, 8-tetrahydrodishridin-4-yl) benzamide

50mg of 3- (5- (2-hydroxyethyl) -7-oxo-5, 6,7, 8-tetrahydrodishrid-4-yl) benzoic acid, 70mg of (S) -2- (2-amino-3- (3-fluorophenyl) propyl) isoindoline-1, 3-dione, 113mg of benzotriazol-1-yloxytris (dimethylamino) phosphonium hexafluorophosphate and 52mg of N, N-diisopropylethylamine were dissolved in 5mL of N, N-dimethylformamide and stirred at room temperature overnight. Pouring the reaction solution into water, separating out solid, filtering, drying, and purifying by column chromatography to obtain 52mg of light yellow solid.

f) (S) -N- (1-amino-3- (3-fluorophenyl) propan-2-yl) -3- (5- (2-hydroxyethyl) -7-oxo-5, 6,7, 8-tetrahydrodishridin-4-yl) benzamide

Dissolving 50mg of the product obtained in the step e in 5mL of methanol, adding 44mg of hydrazine hydrate, stirring at room temperature overnight, evaporating the reaction solution under reduced pressure, and performing column chromatography to obtain 14mg of off-white solid.

LC/MS(ESI+):465.2(M+H)

¹H NMR(400MHz,DMSO-d₆)δ8.53(d,J＝13.5Hz,1H),8.36(d,J＝6.4Hz,1H),8.28(d,J＝8.3Hz,1H),8.14(t,J＝7.3Hz,1H),7.86(d,J＝7.8Hz,1H),7.55(t,J＝7.8Hz,1H),7.30(dd,J＝14.3,7.9Hz,1H),7.15–7.05(m,2H),6.99(dd,J＝11.9,5.4Hz,1H),4.53(s,1H),3.89(d,J＝17.4Hz,2H),3.24–3.09(m,4H),2.69(ddd,J＝21.0,12.9,5.7Hz,4H).

Example 29(S) -N- (1-amino-3- (3-fluorophenyl) propan-2-yl) -3- (5- (cyanomethyl) -7-oxo-5, 6,7, 8-tetrahydronaphthyridin-4-yl) benzamide

Detailed description of the inventionthe title compound was prepared starting from 2- (4-chloro-7-oxo-7, 8-dihydropteridin-5 (6H) -yl) acetonitrile and (3- (methoxycarbonyl) phenyl) boronic acid in accordance with example 4.

LC/MS(ESI+):460.2(M+H)

¹H NMR(400MHz,DMSO-d₆)δ8.52(s,1H),8.39(d,J＝8.4Hz,1H),8.26(s,1H),8.03(d,J＝7.8Hz,1H),7.90(d,J＝7.8Hz,1H),7.55(t,J＝7.7Hz,1H),7.30(dd,J＝14.3,8.1Hz,1H),7.10(d,J＝8.1Hz,2H),6.99(t,J＝8.0Hz,1H),4.17(s,2H),4.06–3.80(m,3H),2.83(dd,J＝26.7,12.9Hz,2H),2.72(t,J＝14.3Hz,2H).

Example 30(S) -N- (1-amino-3- (3-fluorophenyl) propan-2-yl) -3- (7-methyl-8-oxo-8, 9-dihydro-7H-purin-6-yl) benzamide

Detailed description of the inventionthe title compound was prepared starting with 6-chloro-7-methyl-7, 9-dihydro-8H-purin-8-one and (3- (methoxycarbonyl) phenyl) boronic acid, according to example 4.

LC/MS(ESI+):421.1(M+H)

Example 31(S) -N- (1-amino-3- (3-fluorophenyl) propan-2-yl) -3- (5, 5-dimethyl-6-oxo-6, 7-dihydro-5H-pyrrolo [2,3-d ] pyrimidin-4-yl) benzamide

Detailed description of the inventionthe title compound was prepared starting with 4-chloro-5, 5-dimethyl-5, 7-dihydro-6H-pyrrolo [2,3-d ] pyrimidin-6-one and (3- (methoxycarbonyl) phenyl) boronic acid as in example 4.

LC/MS(ESI+):434.2(M+H)

¹H NMR(400MHz,DMSO-d₆)δ8.73(s,1H),8.30(d,J＝8.3Hz,1H),7.93(dd,J＝9.4,5.9Hz,2H),7.66–7.55(m,2H),7.27(dd,J＝14.4,7.8Hz,1H),7.07(t,J＝6.6Hz,2H),6.97(t,J＝8.6Hz,1H),4.17–4.06(m,1H),2.87–2.73(m,2H),2.65(dd,J＝13.5,6.0Hz,2H),1.12(d,J＝8.3Hz,6H).

Example 32N- ((S) -1-amino-3- (3-fluorophenyl) propan-2-yl) -3- (5-methyl-6-oxo-6, 7-dihydro-5H-pyrrolo [2,3-d ] pyrimidin-4-yl) benzamide

Detailed description of the inventionthe title compound was prepared starting with 4-chloro-5-methyl-5, 7-dihydro-6H-pyrrolo [2,3-d ] pyrimidin-6-one and (3- (methoxycarbonyl) phenyl) boronic acid as in example 4.

LC/MS(ESI+):420.2(M+H)

¹H NMR(400MHz,DMSO-d₆)δ8.81(d,J＝1.0Hz,1H),8.53(dd,J＝9.6,6.3Hz,2H),8.29(d,J＝8.7Hz,1H),7.91(d,J＝7.8Hz,1H),7.60(t,J＝7.7Hz,1H),7.39–7.24(m,1H),7.10(q,J＝7.6Hz,2H),7.07–6.87(m,1H),6.76(s,1H),3.03–2.88(m,2H),2.81(dd,J＝13.4,9.0Hz,2H),2.68(d,J＝6.1Hz,2H),1.24(s,3H).

Example 33N- ((S) -1-amino-3- (3-fluorophenyl) propan-2-yl) -3- (5-hydroxy-5-methyl-6-oxo-6, 7-dihydro-5H-pyrrolo [2,3-d ] pyrimidin-4-yl) benzamide

a)3- (5-methyl-6-oxo-6, 7-dihydro-5H-pyrrolo [2,3-d ] pyrimidin-4-yl) benzoic acid methyl ester

250mg of 4-chloro-5-methyl-5, 7-dihydro-6H-pyrrolo [2,3-d ] pyrimidin-6-one, 347mg of (3- (methoxycarbonyl) phenyl) boronic acid, 50mg of [1,1' -bis (diphenylphosphino) ferrocene ] palladium dichloride and 886mg of cesium carbonate are dissolved in a mixed solution of 10mL of dioxane and 1mL of water, the mixture is heated to reflux under the protection of nitrogen, the heating is stopped after 5H, and the reaction solution is evaporated to dryness under reduced pressure and subjected to column chromatography to obtain 200mg of light yellow solid.

LC/MS(ESI+):284.1(M+H).

¹H NMR(400MHz,DMSO-d₆)δ11.63(s,1H),8.80(s,1H),8.53(s,1H),8.20(d,J＝7.8Hz,1H),8.11(d,J＝7.7Hz,1H),7.71(t,J＝7.8Hz,1H),4.26(q,J＝7.6Hz,1H),3.91(s,3H),1.12(d,J＝7.6Hz,3H).

b)3- (5-hydroxy-5-methyl-6-oxo-6, 7-dihydro-5H-pyrrolo [2,3-d ] pyrimidin-4-yl) benzoic acid

Dissolving 200mg of methyl 3- (5-methyl-6-oxo-6, 7-dihydro-5H-pyrrolo [2,3-d ] pyrimidin-4-yl) benzoate in 20mL of methanol, adding 6mL of 2M lithium hydroxide solution, stirring at room temperature for 6H, adding 2M hydrochloric acid to adjust the pH value to 6-7, decompressing and evaporating the reaction solution, and performing column chromatography to obtain 160mg of off-white solid.

LC/MS(ESI+):286.1(M+H).

¹H NMR(400MHz,DMSO-d₆)δ11.73(s,1H),8.81(d,J＝5.2Hz,2H),8.58(d,J＝7.8Hz,1H),8.08(d,J＝7.7Hz,1H),7.65(t,J＝7.8Hz,1H),7.00(s,1H),3.15(s,1H),1.24(s,3H).

c) N- ((S) -1- (1, 3-dioxoisoindolin-2-yl) -3- (3-fluorophenyl) propan-2-yl) -3- (5-hydroxy-5-methyl-6-oxo) -6, 7-dihydro-5H-pyrrolo [2,3-d ] pyrimidin-4-yl) benzamide

160mg of 3- (5-hydroxy-5-methyl-6-oxo-6, 7-dihydro-5H-pyrrolo [2,3-d ] pyrimidin-4-yl) benzoic acid, 240mg of (S) -2- (2-amino-3- (3-fluorophenyl) propyl) isoindoline-1, 3-dione, 391mg of benzotriazol-1-yloxytris (dimethylamino) phosphonium hexafluorophosphate and 119mg of N, N-diisopropylethylamine were dissolved in 10mL of N, N-dimethylformamide and stirred at room temperature overnight. Pouring the reaction solution into ethyl acetate, washing with water, taking an organic phase, evaporating to dryness under reduced pressure, and purifying by column chromatography to obtain 180mg of off-white solid. LC/MS (ESI +):566.1(M + H).

d) N- ((S) -1-amino-3- (3-fluorophenyl) propan-2-yl) -3- (5-hydroxy-5-methyl-6-oxo-6, 7-dihydro-5H-pyrrolo [2,3-d ] pyrimidin-4-yl) benzamide

180mg of N- ((S) -1- (1, 3-dioxoisoindolin-2-yl) -3- (3-fluorophenyl) propan-2-yl) -3- (5-hydroxy-5-methyl-6-oxo) -6, 7-dihydro-5H-pyrrolo [2,3-d ] pyrimidin-4-yl) benzamide are dissolved in 20mL of methanol, 328mg of 80% hydrazine hydrate are added, the mixture is stirred overnight at room temperature, the reaction mixture is evaporated to dryness under reduced pressure, and column chromatography is carried out to obtain 80mg of an off-white solid.

LC/MS(ESI+):436.1(M+H).

¹H NMR(400MHz,DMSO-d₆)δ8.82(s,1H),8.54(ddd,J＝9.9,7.0,2.2Hz,2H),8.31(d,J＝8.4Hz,1H),7.97–7.83(m,1H),7.60(t,J＝7.7Hz,1H),7.30(ddd,J＝14.4,8.0,6.5Hz,1H),7.10(ddd,J＝10.1,7.6,5.5Hz,2H),6.98(tt,J＝8.5,3.1Hz,1H),4.14(tt,J＝10.9,8.5,4.1Hz,1H),2.96(ddd,J＝13.7,5.5,2.0Hz,1H),2.90–2.75(m,1H),2.77–2.56(m,2H),2.48(s,2H),1.24(s,3H).

Example 34N- ((S) -1-amino-3- (3-fluorophenyl) propan-2-yl) -3- ((R) -5-hydroxy-5-methyl-6-oxo-6, 7-dihydro-5H-pyrrolo [2,3-d ] pyrimidin-4-yl) benzamide and N- ((S) -1-amino-3- (3-fluorophenyl) propan-2-yl) -3- ((S) -5-hydroxy-5-methyl-6-oxo-6, 7-dihydro-5H-pyrrolo [2,3-d ] pyrimidin-4-yl) benzamide

The product of example 33 was resolved on a chiral column to give the title compound.

Chiral resolution conditions:

the instrument comprises the following steps: waters SFC; a chiral column, Daicel Chiralcel AD,250 × 30mm I.D.,5 μm; the mobile phase A is carbon dioxide, and the phase B is ethanol (containing 0.1% ammonia water); a: b is 60: 40 (volume ratio); the flow rate is 60 mL/min; the column temperature was 38 ℃. The product was collected with retention time 13-18min as isomer 1(24mg) and the product was collected with retention time 27-41min as isomer 2(35 mg).

Conditions for HPLC analysis:

the instrument comprises the following steps: waters UPCC, chiral column: daicel Chiralcel AD,2.1 × 150mm i.d.,3 μm, mobile phase: phase A is CO₂Phase B ethanol (containing 0.1% diethanolamine), gradient elution: 0-6.5min, phase B: 5-40% (volume ratio), flow rate: 1mL/min, column temperature: at 40 ℃.

Isomer 1: RT 5.3min, LC/MS (ESI +):436.1(M + H)

Isomer 2: RT 6.5min, LC/MS (ESI +):436.1(M + H)

Example 35(S) -N- (1-amino-3- (3, 5-difluorophenyl) propan-2-yl) -3- (5-methyl-7-oxo-5, 6,7, 8-tetrahydrodishridin-4-yl) benzamide

Specifically, referring to example 4, the title compound was prepared by using 4-chloro-5-methyl-5, 8-dihydropteridin-7 (6H) -one and (3-methoxycarbonyl) phenyl) boronic acid as raw materials to prepare methyl 3- (5-methyl-7-oxo-5, 6,7, 8-tetrahydropteridin-4-yl) benzoate, hydrolyzing the methyl 3- (5-methyl-7-oxo-5, 6,7, 8-tetrahydropteridin-4-yl) benzoate, reacting the hydrolyzed methyl with (S) -2- (2-amino-3- (3, 5-difluorophenyl) propyl) isoindoline-1, 3-dione, and deprotecting the resulting product.

LC/MS(ESI+):453.2(M+H).

¹H NMR(400MHz,CDCl₃)δ8.61(d,J＝3.3Hz,1H),8.40(d,J＝24.0Hz,1H),8.18(d,J＝8.0Hz,1H),7.94–7.72(m,2H),7.55(dd,J＝15.4,7.8Hz,2H),6.92–6.74(m,2H),6.68(t,J＝9.0Hz,1H),4.36(d,J＝6.6Hz,1H),3.89(s,2H),3.04(m,1H),2.96–2.79(m,2H),2.54(s,3H),2.27–2.15(m,1H),2.02(m,1H).

Example 36N- (1-amino-3- (3, 4-difluorophenyl) propan-2-yl) -3- (5-methyl-7-oxo-5, 6,7, 8-tetrahydrodishridin-4-yl) benzamide

Specifically, referring to example 4, the title compound was prepared by using 4-chloro-5-methyl-5, 8-dihydropteridin-7 (6H) -one and (3-methoxycarbonyl) phenyl) boronic acid as starting materials to prepare methyl 3- (5-methyl-7-oxo-5, 6,7, 8-tetrahydropteridin-4-yl) benzoate, hydrolyzing the methyl 3- (5-amino-3- (3, 5-difluorophenyl) propyl) isoindoline-1, 3-dione, and deprotecting the reaction.

LC/MS(ESI+):453.2(M+H)

Test example 1 in vitro enzyme Activity test

1. Experimental procedure

A Mobility shift assay method is adopted to establish an AKT1, AKT2 and AKT3 kinase activity detection platform, and the activity of the compound is determined. Starting from 100. mu.M, the compound was reacted with 100% DMSO (purchased from Sigma, Cat: D8418-1L) was diluted 3-fold in gradient (10 concentrations in total) and 250nL of compound was transferred to the destination plate OptiPlate-384F (purchased from Perkinelmer, Cat: 6007290) using a dispenser Echo 550 for each concentration. Positive and negative control wells were loaded with 250nL DMSO. Add 10. mu.L of AKT kinase solution (purchased from Carna, AKT1 Cat:01-101, AKT2 Cat:01-102, AKT3 Cat: 01-103; AKT1 kinase final concentration 0.5nM, AKT2 kinase final concentration 0.05nM, AKT3 kinase final concentration 0.25nM) to the compound wells and the positive control wells, respectively; mu.L of 1 XKinase buffer (containing 50mM HEPES, 10mM MgCl. RTM.) was added to the negative control wells₂2mM DTT). Then, the mixture was centrifuged at 1000rpm for 30 seconds, shaken and mixed, and then incubated at room temperature for 10 minutes. A mixture of 15. mu.L ATP (purchased from Sigma, Cat. A7699-5G, Lot. SLBT6850, AKT1 at a final ATP concentration of 120. mu.M, AKT2 at a final ATP concentration of 500. mu.M, and AKT3 at a final ATP concentration of 29.3. mu.M) and Caliper substrate 6 (purchased from GL, Cat: 116368 at a final substrate concentration of 3. mu.M) was added to initiate a reaction, and the total reaction volume was 25. mu.L. And (3) centrifuging the 384-well plate at 1000rpm for 30 seconds, shaking and uniformly mixing, and then respectively incubating at room temperature for corresponding time (60 min for AKT1 detection, 30min for AKT2 detection and 10min for AKT3 detection). The kinase reaction was stopped by adding 30. mu.L of a termination test solution (termination test solution containing 50mM EDTA), and the mixture was centrifuged at 1000rpm for 30 seconds and shaken and mixed. The conversion was read at Caliper EZ Reader II (equipment parameters: -1.2PSI, upstream voltage-500, downstream voltage-2250, reading interval 45 s). The IC of the compound was obtained by fitting a dose-response curve to GraphPad Prism 5 software log (inhibitor) vs. response-Variable slope₅₀The value is obtained.

Inhibition rate calculation formula:

wherein:

conversion% _ sample: is the conversion reading for the sample;

conversion% _ min: negative control well mean, representing conversion readings without enzyme live wells;

conversion% _ max: positive control well mean, representing conversion readings for wells without compound inhibition.

2. Results of the experiment

The results of the experiments are shown in the following table:

Claims

1. a compound of formula I or a pharmaceutically acceptable salt thereof,

wherein:

R¹and R²Each independently selected from H or C1-C6 alkyl;

m is selected from 0 or 1;

is selected from

l is

A group wherein:

Y¹¹and Y²¹Each independently selected from H, OH, halogen or C1-C6 alkyl, wherein C1-C6 alkyl may be optionally substituted with halogen, OH, C1-C3 alkoxy or CN;

Y¹²and Y²²Each independently selected from H or C1-C6 alkyl;

2. A compound of formula I according to claim 1, wherein:

R¹and R²Each independently selected from H, methyl, ethyl or isopropyl; preferably, R¹Selected from H, R²Selected from H, methyl, ethyl or isopropyl;

more preferably, R¹Selected from H, R²Selected from H or isopropyl; most preferably, R¹Selected from H, R²Is selected from H;

m is selected from 1;

a is substituted by 1 or 2R³Substituted phenyl; preferably, a is selected from the following groups:

R³independently selected from F, Cl, Br, I, CH₃、C₂H₅Or CF₃(ii) a Preferably, R³Selected from F, Cl or CF₃(ii) a More preferablyEarth, R³Selected from F or Cl; most preferably, R³Is selected from F.

3. A compound of formula I according to claim 1, wherein:

a is selected from the following groups:

preferably, a is selected from the following groups:

more preferably, a is selected from the following groups:

most preferably, A is selected from

4. A compound of formula I according to claim 1, wherein:

is selected from

5. A compound of formula I according to claim 1, wherein:

g is selected from: a five-membered heteroaryl, pyridyl or phenyl group, wherein the five-membered heteroaryl, pyridyl or phenyl group is optionally substituted with R⁴Is substituted, and R⁴Selected from halogen or C1-C6 alkyl; preferably, R⁴Is selected from CH₃F or Cl; more preferably, R⁴Is selected from CH₃Or F;

preferably, G is selected from optionally substituted with R⁴Substituted of the following groups:

wherein,

represents the position where G is attached to Q by a chemical bond,

represents the position at which G is attached to L by a chemical bond;

wherein R is⁴As defined above.

6. The compound of formula I according to claim 5, wherein:

g is selected from optionally substituted with R⁴Substituted of the following groups:

more preferably, G is selected from the following groups:

further preferably, G is selected from the following groups:

most preferably, G is selected from the following groups:

7. a compound of formula I according to claim 1, wherein:

Y¹is selected from (a): CY¹¹Y¹²Wherein Y is¹¹Selected from H, OH or C1-C6 alkyl, wherein C1-C6 alkyl may be optionally substituted with F; preferably, Y¹¹Selected from H, CH₃Or CF₃(ii) a More preferably, Y¹¹Is selected from CH₃Or CF₃(ii) a Most preferably, Y¹¹Is selected from CH₃；，Y¹²Is selected from H or CH₃；

Y¹Is selected from (b): NY¹³Wherein Y is¹³Selected from H, CH₃、C₂H₅、CH₂CH₂CH₃、CHCH₃CH₃、CH₂OH、C₂H₄OH、C₃H₆OH、CH₂OCH₃、CH₂OC₂H₅、C₂H₄OCH₃、CH₂CN or C₂H₄CN; preferably, Y¹³Is selected from CH₃、C₂H₄OH or CH₂CN; more preferably, Y¹³Is selected from CH₃；

Y²Selected from O, CY²¹Y²²Or a bond, wherein Y²¹And Y²²Are all H; preferably, Y²Is selected from CH₂Or a key.

8. A compound of formula I according to claim 7, L is selected from the following groups:

preferably, G is selected from the following groups:

more preferably, G is selected from the following groups:

9. the present invention provides the following compounds or pharmaceutically acceptable salts thereof: