CN116157396A - Pyridazinone derivatives and their applications in medicine - Google Patents

Abstract

Pyridazinone derivatives are provided, as are PARP inhibitors, such as PARP-7 inhibitors, and their use in medicine.

Description

Pyridazinone derivatives and their application in medicine technical field

The present invention relates to pyridazinone derivatives or their stereoisomers or pharmaceutically acceptable salts and their application in medicine.

Background technique

Adenosine diphosphate-ribosylation (ADP-ribosylation, ADP-ribosylation) is a protein post-transcriptional modification process, in which single or multiple adenosine diphosphate-ribose (ADP-ribose) groups are embedded in amino acid residues of proteins. ADP-ribosylation is a reversible process involved in physiological regulation of cell signal transduction, DNA damage repair, transcription, regulation of gene expression, and apoptosis. ADP-ribose is derived from a redox cofactor: Nicotinamide adenine dinucleotide (NAD+), and the enzyme that mediates the insertion modification of ADP-ribose is ADP-ribosylase. In this physiological response regulation, the N-glycosidic bond of NAD+ connecting the ADP-ribose molecule and the nicotinamide group is cleaved, and then it is captured and bonded with the corresponding amino acid residue of the target protein. ADP-ribosylases can perform two types of modifications: mono-ADP ribosylation and poly-ADP ribosylation. When DNA is damaged or cells are stressed, PARP is activated, resulting in an increase in the amount of poly ADP-ribose and a decrease in the amount of NAD+. For more than a decade, PARP1 was thought to be the only poly-ADP-ribose polymerase in mammalian cells and thus the most studied enzyme. To date, scientists have identified 17 different PARPs. MonoPARP occupies the majority of the PARP family and mediates important biological functions and various stress responses, such as: unfolded protein response, NF-κB signaling, antiviral response, and cytokine signaling. 2,3,7,8-Tetrachlorodibenzo-p-dioxin (TCDD)-inducible poly(ADP-ribose) polymerase (PARP-7) is a member of the MonoPARP family, and its expression is activated by TCDD. Regulation of AHR (AHR), a ligand-activated transcription factor that mediates the toxic activity of many environmental xenobiotics. AHR up-regulates the expression of PARP-7, and PARP-7 interacts with the kinase TBK1 and makes it ADP-ribosylated, leading to the inhibition of TBK1 activity and the down-regulation of IFN-I (type I interferon) response, which in turn leads to the body's antiviral and tumor immune responses are suppressed.

Contents of the invention

It is an object of one or more embodiments of the present application to provide novel PARP inhibitors.

One or more embodiments of the present application aim to provide novel PARP-7 inhibitors.

One or more embodiments of the present application provide a compound of general formula (I), or a stereoisomer or a pharmaceutically acceptable salt thereof:

in

R, R', R" are each independently H or C _1-6 alkyl; or

R and R' or R and R" form a 4 to 8-membered heterocycloalkyl with the connected atoms;

R ₁ and R ₂ are each independently H or C _1-6 alkyl;

L ₁ is a bond or C=O;

L is a bond or NH;

C ₁ is 6-membered, 7-membered or 8-membered heterocyclic group;

R ₃ and R ₄ are each independently H, halogen or C _1-6 alkyl, wherein the C _1-6 alkyl is optionally further substituted by 1 or more substituents selected from OH or CN;

C is a 6-membered heteroaryl, and the 6-membered heteroaryl is optionally further substituted by 1 or more substituents selected from CN or C _1-6 alkyl optionally substituted by 1 or more halogens ;

C ₂ is C _3-6 carbocyclic ring;

_Each R is independently H, halogen, CN or OH;

n is 1 or 2;

m is 0, 1, 2, 3, 4 or 5.

In one or more embodiments, _C is a 6-, 7- or 8-membered heterocyclyl comprising 1, 2 or 3 N atoms.

In one or more embodiments, C _is

In one or more embodiments, C is a 6-membered heteroaryl group containing 1, 2 or 3 N atoms, and the 6-membered heteroaryl group is optionally further selected from CN or any is optionally substituted with 1 or more halogen-substituted C _1-6 alkyl substituents.

其任选地进一步被1或多个选自CN或者任选地被1或多个卤素取代的C _1-6烷基的取代基所取代。 In one or more embodiments, C is

It is optionally further substituted with 1 or more substituents selected from CN or C _1-6 alkyl optionally substituted with 1 or more halogen.

In one or more embodiments, the heterocycloalkyl group is a 4-membered, 5-membered or 6-membered heterocycloalkyl group containing 1 N atom.

In one or more embodiments, R is H.

In one or more embodiments, R', R" are each independently H or C _1-6 alkyl;

In one or more embodiments, _R and _R are H.

In one or more embodiments, L ₁ is C=O.

In one or more embodiments, L is a bond.

In one or more embodiments, _L1 is C=O and L is NH.

In one or more embodiments, L ₁ is C=O and L is a bond.

In one or more embodiments, C _is

In one or more embodiments, each of R ₃ and R ₄ is independently halogen or C _1-6 alkyl, wherein the C _1-6 alkyl is optionally further substituted with 1 or more OH.

In one or more embodiments, C is

In one or more embodiments, _C2 is a _C3 carbocycle.

In one or more embodiments, R ₅ is halogen.

In one or more embodiments, n is 2.

In one or more embodiments, m is zero.

One or more embodiments of the present application provide a compound of general formula (II), or a stereoisomer or a pharmaceutically acceptable salt thereof:

in

R ₁ and R ₂ are each independently selected from H or C _1-6 alkyl;

R ₃ and R ₄ are each independently selected from H, halogen or C _1-6 alkyl, wherein the C _1-6 alkyl is optionally further substituted by 1 or more substituents selected from OH or CN;

_Each R is independently selected from H, halogen, CN or OH;

L is a bond or -NH-;

C ₁ for

C ₂ is C _3-5 carbocyclic ring;

n is 1 or 2;

m is 0, 1, 2, 3, 4 or 5.

In one or more embodiments, _R and _R are H.

In one or more embodiments, L is a bond.

In one or more embodiments, each of R ₃ and R ₄ is independently halogen or C _1-6 alkyl, wherein the C _1-6 alkyl is optionally further substituted with 1 or more OH.

In one or more embodiments, _C2 is a _C3 carbocycle.

In one or more embodiments, R ₅ is halogen.

In one or more embodiments, n is 2.

In one or more embodiments, m is zero.

One or more embodiments of the present application provide a compound or a stereoisomer or a pharmaceutically acceptable salt thereof, which is selected from the following structures:

One or more embodiments of the present application provide intermediate compounds or stereoisomers or pharmaceutically acceptable salts thereof for the preparation of compounds of general formula (I), (II) or their stereoisomers or pharmaceutically acceptable salts Acceptable salts, which have compounds represented by general formula (III), (IV), (V) or (VI):

in

R, R', R" are each independently H or C _1-6 alkyl; or

R and R' or R and R" form a 4 to 8-membered heterocycloalkyl with the connected atoms;

R ₁ and R ₂ are each independently H or C _1-6 alkyl;

L ₁ is a bond or C=O;

L is a bond or NH;

C is ₆ to 8 membered heterocyclic groups;

R ₃ and R ₄ are each independently H, halogen or C _1-6 alkyl, wherein the C _1-6 alkyl is optionally further substituted by 1 or more substituents selected from OH or CN;

C is a 6-membered heteroaryl group, and the 6-membered heteroaryl group is optionally further substituted by 1 or more substituents selected from CN or C _1-6 alkyl, and the C _1-6 alkyl is optionally is further substituted by 1 or more halogens;

C ₂ is C _3-6 carbocyclic ring;

_Each R is independently H, halogen, CN or OH;

C ₃ is a 6-8 membered heterocyclic group;

X ₁ is H or NH ₂ ;

n is 1 or 2;

m is 0, 1, 2, 3, 4 or 5.

In one or more embodiments, the heterocycloalkyl group is a 4-membered, 5-membered or 6-membered heterocycloalkyl group containing 1 N atom.

In one or more embodiments, _C is a 6-, 7- or 8-membered heterocyclyl comprising 1, 2 or 3 N atoms.

In one or more embodiments, C is a 6-membered heteroaryl group comprising 1, 2 or 3 N atoms.

In one or more embodiments, C _is

其任选地进一步被1或多个选自CN或C _1-6烷基的取代基所取代，所述C _1-6烷基任选地进一步被1或多个卤素所取代。 In one or more embodiments, C is

It is optionally further substituted by 1 or more substituents selected from CN or C _1-6 alkyl, and said C _1-6 alkyl is optionally further substituted by 1 or more halogen.

In one or more embodiments, _C3 is

In one or more embodiments, the intermediate compound or its stereoisomer or pharmaceutically acceptable salt has the following structure or its stereoisomer or pharmaceutically acceptable salt:

One or more embodiments of the present application provide a pharmaceutical composition comprising:

(1) The compound of the present application, or its stereoisomer or pharmaceutically acceptable salt;

(2) Optional one or more other active ingredients; and

(3) Pharmaceutically acceptable carriers and/or excipients.

One or more embodiments of the present application provide the use of the compound of the present application or its stereoisomer or pharmaceutically acceptable salt or the pharmaceutical composition of the present application in the preparation of antitumor drugs.

In one or more embodiments, said tumor formation is associated with PARP.

In one or more embodiments, the PARP is PARP-7.

One or more embodiments of the present application provide the use of the compound of the present application or its stereoisomer or pharmaceutically acceptable salt or the pharmaceutical composition of the present application in the preparation of a PARP inhibitor.

In one or more embodiments, the PARP is PARP-7.

One or more embodiments of the present application provide the compound of the present application or its stereoisomer or pharmaceutically acceptable salt or the pharmaceutical composition of the present application for use as a medicament.

One or more embodiments of the present application provide the compound of the present application or its stereoisomer or pharmaceutically acceptable salt or the pharmaceutical composition of the present application, which is used in the method of treating tumor. In one or more embodiments, said tumor formation is associated with PARP. In one or more embodiments, the PARP is PARP-7.

One or more embodiments of the present application provide the compound of the present application or its stereoisomer or pharmaceutically acceptable salt or the pharmaceutical composition of the present application, which is used as a PARP inhibitor, such as a PARP-7 inhibitor.

One or more embodiments of the present application provide a method for treating tumors, which includes administering the compound of the present application or its stereoisomer or pharmaceutically acceptable salt or the pharmaceutical composition of the present application to a subject in need thereof.

One or more embodiments of the present application provide a method for inhibiting PARP, which includes administering the compound of the present application or its stereoisomer or pharmaceutically acceptable salt or the pharmaceutical composition of the present application. In one or more embodiments, the PARP is PARP-7.

Detailed description of the invention

Unless stated to the contrary, the terms used in the specification and claims have the following meanings.

The carbon, hydrogen, oxygen, sulfur, nitrogen or F, Cl, Br, and I involved in the groups and compounds of the present invention include their isotopes, and the carbon involved in the groups and compounds of the present invention , hydrogen, oxygen, sulfur or nitrogen are optionally further replaced by one or more of their corresponding isotopes, wherein the isotopes of carbon include ¹² C, ¹³ C and ¹⁴ C, and the isotopes of hydrogen include protium (H), deuterium (D, Also called heavy hydrogen), tritium (T, also called super heavy hydrogen), oxygen isotopes include ¹⁶ O, ¹⁷ O and ¹⁸ O, sulfur isotopes include ³² S, ³³ S, ³⁴ S and ³⁶ S, nitrogen isotopes include ¹⁴ N and ¹⁵ N, the isotopes of fluorine include ¹⁷ F and ¹⁹ F, the isotopes of chlorine include ³⁵ Cl and ³⁷ Cl, and the isotopes of bromine include ⁷⁹ Br and ⁸¹ Br.

"Alkyl" means 1 to 20 carbon atoms (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19 or 20 carbon atoms) straight-chain or branched saturated aliphatic hydrocarbon group, preferably an alkyl group of 1 to 8 carbon atoms, more preferably an alkyl group of 1 to 6 carbon atoms, further preferably 1 to 4 Alkyl group of carbon atoms. Non-limiting examples include methyl, ethyl, n-propyl, isopropyl, n-butyl, sec-butyl, neobutyl, tert-butyl, n-pentyl, isopentyl, neopentyl, n-hexyl And its various branched isomers; when the alkyl group is substituted, it may be optionally further substituted by one or more substituents.

"Aryl" refers to a substituted or unsubstituted aromatic ring, which can be a 5- to 8-membered (eg 5, 6, 7, 8-membered) monocyclic ring, a 5- to 12-membered (eg 5, 6, 7-membered) , 8, 9, 10, 11, 12 member) bicyclic ring or 10 to 15 member (eg 10, 11, 12, 13, 14, 15 member) tricyclic ring system, which can be a bridge ring or a spiro ring, non-limiting implementation Examples include phenyl, naphthyl. The aryl group may be optionally further substituted by one or more substituents.

"Heteroaryl" refers to a substituted or unsubstituted aromatic ring, which can be a 5 to 8 membered (eg 5, 6, 7, 8 membered) monocyclic ring, a 5 to 12 membered (eg 5, 6, 7, 8, 9, 10, 11, 12 member) bicyclic ring or 10 to 15 member (eg 10, 11, 12, 13, 14, 15 member) tricyclic ring system, and contains 1 to 6 (eg 1, 2, 3, 4, 5, 6) heteroatoms selected from N, O or S, preferably 3 to 8-membered (eg 3, 4, 5, 6, 7, 8-membered) heterocyclic group, the heterocyclic group is optional in the ring Substituted N and S can be oxidized into various oxidation states. The heterocyclic group can be connected to a heteroatom or a carbon atom, and the heterocyclic group can be a bridge ring or a spiro ring. Non-limiting examples include cyclopyridyl, furyl, thienyl, pyryl, pyrrolyl, pyrimidinyl, Pyrazinyl, pyridazinyl, imidazolyl, piperidylbenzimidazolyl, benzopyridyl, pyrrolopyridyl. When heteroaryl is substituted, it may be optionally further substituted with one or more substituents.

氮杂二环[5.2.0]壬烷基、氧杂三环[5.3.1.1]十二烷基、氮杂金刚烷基和氧杂螺[3.3]庚烷基。所述的“杂环基”或“杂环”可以任选进一步被0个或者多个取代基所取代。 "Heterocyclic group" or "heterocyclic ring" refers to a saturated or unsaturated heteroaromatic or non-heteroaromatic ring. When selected from a heteroaromatic ring, its definition is the same as the definition of "heteroaryl"above; In the case of an aromatic ring, it may be a monocyclic ring of 3 to 10 members (for example, 3, 4, 5, 6, 7, 8, 9, 10 members), a 4 to 12 member (for example, 4, 5, 6, 7, 8, 9, 10, 11, 12) bicyclic or 10 to 15 (e.g. 10, 11, 12, 13, 14, 15) tricyclic ring system, and contains 1 to 4 (e.g. 1, 2, 3, 4 ) is a heteroatom selected from N, O or S, preferably a 3- to 8-membered heterocyclic group. The selectively substituted N and S in the ring of "heterocyclic group" or "heterocyclic ring" can be oxidized to various oxidation states; "heterocyclic group" or "heterocyclic ring" can be connected to a heteroatom or a carbon atom; " A "heterocyclyl" or "heterocycle" may be a bridged ring or a spiro ring. Non-limiting examples of "heterocyclyl" or "heterocycle" include oxiranyl, epoxypropyl, aziridyl, oxetanyl, azetidinyl, thietanyl , 1,3-dioxolanyl, 1,4-dioxolanyl, 1,3-dioxanyl, azepanyl, oxepyl, thiepanyl, oxygen Azepine, diazepine, thiazepine, pyridyl, piperidyl, homopiperidyl, furyl, thienyl, pyryl, N-alkylpyrrolyl, pyrimidyl, pyrimidyl Azinyl, pyridazinyl, piperazinyl, homopiperazinyl, imidazolyl, piperidinyl, morpholinyl, thiomorpholinyl, thioxanyl, 1,3-dithianyl, dihydrofuran base, dithiapentyl, tetrahydrofuranyl, tetrahydrothiophenyl, tetrahydropyranyl, tetrahydrothiopyranyl, tetrahydropyrrolyl, tetrahydroimidazolyl, tetrahydrothiazolyl, tetrahydropyranyl, benzene Imidazolyl, benzopyridyl, pyrrolopyridyl, chromanyl, 2-pyrrolinyl, 3-pyrrolinyl, indolinyl, 2H-pyranyl, 4H-pyranyl , Dioxanyl, 1,3-dioxolyl, pyrazolinyl, dithianyl, dithianyl, dihydrothienyl, pyrazolidinyl, imidazolinyl, imidazolidinyl, 1,2,3,4-tetrahydroisoquinolinyl, 3-azabicyclo[3.1.0]hexyl, 3-azabicyclo[4.1.0]heptyl, azabicyclo[2.2.2]hexyl, 3H-indolylquinazinyl, N-pyridylurea, 1,1-dioxothiomorpholinyl, azabicyclo[3.2.1]octyl,

Azabicyclo[5.2.0]nonyl, oxatricyclo[5.3.1.1]dodecyl, azaadamantyl and oxaspiro[3.3]heptanyl. The "heterocyclic group" or "heterocycle" may be optionally further substituted by 0 or more substituents.

"Heterocycloalkyl" refers to a substituted or unsubstituted saturated non-aromatic ring group, which may be 3 to 8 membered (eg 3, 4, 5, 6, 7 or 8 membered) monocyclic, 4 to 12 membered (e.g. 4, 5, 6, 7, 8, 9, 10, 11, 12 members) bicyclic rings or 10 to 15 membered (e.g. 10, 11, 12, 13, 14, 15 members) tricyclic ring systems, and comprising 1, 2 or 3 heteroatoms selected from N, O or S, eg 3 to 8 membered heterocyclyl. The N and S selectively substituted in the ring of "heterocycloalkyl" can be oxidized into various oxidation states; "heterocycloalkyl" can be connected to a heteroatom or carbon atom; "heterocycloalkyl" can be a bridge ring or spiral. Non-limiting examples of "heterocycloalkyl" include oxiranyl, aziridinyl, oxetanyl, azetidinyl, 1,3-dioxolanyl, 1,4-di Oxolanyl, 1,3-dioxanyl, azepanyl, piperidinyl, piperidinyl, morpholinyl, thiomorpholinyl, 1,3-dithianyl, tetrahydrofuranyl , tetrahydropyrrolyl, tetrahydroimidazolyl, tetrahydrothiazolyl, tetrahydropyranyl, azabicyclo[3.2.1]octanyl, azabicyclo[5.2.0]nonyl, oxa Tricyclo[5.3.1.1]dodecyl, azaadamantyl and oxaspiro[3.3]heptanyl.

所述的“碳环基”或“碳环”可以任选进一步被1个或者多个取代基所取代。 "Carbocyclyl" or "carbocycle" refers to a saturated or unsaturated aromatic or non-aromatic ring. When selected from an aromatic ring, its definition is the same as the definition of "aryl"above; when selected from a non-aromatic ring, it can be a 3-10-membered monocyclic ring, a 4-12-membered bicyclic ring, or a 10-15-membered tricyclic ring system , can be bridged or spiro, non-limiting examples include cyclopropyl, cyclobutyl, cyclopentyl, 1-cyclopentyl-1-enyl, 1-cyclopentyl-2-enyl, 1 -Cyclopentyl-3-enyl, cyclohexyl, 1-cyclohexyl-2-enyl, 1-cyclohexyl-3-enyl, cyclohexenyl, cyclohexadienyl, cycloheptyl, cyclooctyl base, cyclononyl, cyclodecyl, cycloundecyl, cyclododecyl,

The "carbocyclyl" or "carbocycle" may be optionally further substituted by one or more substituents.

When the above-mentioned "alkyl", "aryl", "heteroaryl", "heterocycle", "heterocyclyl", "heterocycloalkyl", "carbocycle", "carbocyclyl" When substituted, it can optionally be further 0, 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 selected from F, Cl, Br, I, hydroxyl, mercapto, nitro, cyano , amino, C _1-6 alkylamino, =O, C _1-6 alkyl, C _1-6 alkoxy, C _2-6 alkenyl, C _2-6 alkynyl, -NR ^q4 R ^q5 , = NR ^q6 , -C(=O)OC _1-6 alkyl, -OC(=O)C _1-6 alkyl, -C(=O)NR ^q4 R ^q5 , C _3-8 cycloalkyl, C _{3 -8} heterocycloalkyl, C _6-10 aryl, C _5-10 heteroaryl, -C(=O)OC _6-10 aryl, -OC(=O)C _6-10 aryl, -OC (=O)C _5-10 heteroaryl, -C(=O)OC _5-10 heteroaryl, -OC(=O)C _3-8 heterocycloalkyl, -C(=O)OC _{3- 8} Heterocycloalkyl, -OC(=O)C _3-8 Cycloalkyl, -C(=O)OC _3-8 Cycloalkyl, -NHC(=O)C _3-8 Heterocycloalkyl, - NHC(=O)C _6-10 aryl, -NHC(=O)C _5-10 heteroaryl, -NHC(=O)C _3-8 cycloalkyl, -NHC(=O)C _3-8 Heterocycloalkyl, -NHC (=O) C _2-6 alkenyl or -NHC (= O) C _2-6 alkynyl is substituted by substituents, and wherein the substituent C _1-6 alkyl, C _1-6 alkoxy, C _2-6 alkenyl, C _2-6 alkynyl, C _3-8 cycloalkyl, C _3-8 heterocycloalkyl, C _6-10 aryl, C _5-10 Heteroaryl, -NHC(=O)C _6-10 aryl, -NHC(=O)C _5-10 heteroaryl, -NHC(=O)C _3-8 heterocycloalkyl or -NHC(= O) C _3-8 cycloalkyl is optionally further replaced by 1 to 3 selected from OH, F, Cl, Br, I, C _1-6 alkyl, C _1-6 alkoxy, -NR ^q4 R ^q5 or =O substituent; R ^q1 is selected from C _1-6 alkyl, C _1-6 alkoxy or C _6-10 aryl; R ^q2 , R ^q3 are selected from H or C _1-6 alkyl; R ^q4 and R ^q5 are selected from H, C _1-6 alkyl, -NH(C=NR ^q1 )NR ^q2 R ^q3 , -S(=O) ₂ NR ^q2 R ^q3 , -C(=O)R ^q1 or -C(=O)NR ^q2 R ^q3 , wherein the C _1-6 alkyl is optionally further replaced by one or more selected from OH, F, Cl, Br, I, C _1-6 alkyl, C Substituents of _1-6 alkoxy, C _6-10 aryl, C _5-10 heteroaryl, C _3-8 cycloalkyl or C _3-8 heterocycloalkyl; or R ^q4 and R ^q5 and N atoms form a 3-8 membered heterocyclic ring, and the ring may contain 1 or more heteroatoms selected from N, O or S.

"Pharmaceutical composition" refers to a mixture of one or more compounds described in the present invention, their pharmaceutically acceptable salts or prodrugs and other active components, wherein "other active components" refers to pharmaceutically acceptable acceptable carrier, excipient and/or one or more other therapeutic agents.

"Carrier" refers to a material that does not produce significant irritation to an organism and that does not abrogate the biological activity and properties of the administered compound.

"Excipient" refers to an inert substance added to a pharmaceutical composition to facilitate administration of the compound. Non-limiting examples include calcium carbonate, calcium phosphate, sugars, starches, cellulose derivatives (including microcrystalline cellulose), gelatin, vegetable oils, polyethylene glycols, diluents, granulating agents, lubricants, binders agents and disintegrants.

"Stereoisomer" refers to isomers produced by different arrangements of atoms in a molecule in space, including cis-trans isomers, enantiomers and conformational isomers.

"Pharmaceutically acceptable salt" or "pharmaceutically acceptable salt thereof" means that the compound of the present invention maintains the biological effectiveness and characteristics of free acid or free base, and the free acid is mixed with a non-toxic inorganic base or Organic base, the salt obtained by reacting the free base with a non-toxic inorganic acid or organic acid.

"Optional" or "optionally" or "optional" or "optionally" means that the subsequently described event or circumstance may, but need not, occur, and that the description includes the circumstances in which the event or circumstance occurs and the circumstances in which it is not what happened. For example, "heterocyclyl optionally substituted with an alkyl group" means that the alkyl group may but need not be present, and the description includes cases where the heterocyclyl group is substituted with an alkyl group, and cases where the heterocyclyl group is not substituted with an alkyl group. Condition.

Detailed ways

The following examples illustrate the technical solution of the present invention in detail, but the protection scope of the present invention includes but is not limited thereto.

PMB: means p-methoxybenzyl.

Intermediate 1

(S)-3-(2-((6-Oxo-5-(trifluoromethyl)-1,6-dihydropyridazin-4-yl)amino)propoxy)propanoic acid (Intermediate 1 )

(S)-3-(2-((6-oxo-5-(trifluoromethyl)-1,6-dihydropyridazin-4-yl)amino)propoxy)propanoic acid

first step:

4,5-Dibromo-2-(4-methoxybenzyl)pyridazin-3(2H)-one (1b)

4,5-dibromo-2-(4-methoxybenzyl)pyridazin-3(2H)-one

4,5-Dibromo-2,3-dihydropyridazin-3-one (1a, 50g, 196.94mmol, 1.0equiv) in N,N-dimethylformamide (500mL ) solution, sodium hydrogen (11.82g, 295.41mmol, 1.5equiv, 60%) was added in batches, and 1-(chloromethyl)-4-methoxybenzene (46.06g, 294.11mmol, 1.49 equiv). After the addition was complete, the reaction mixture was stirred at room temperature for 3 h. After the reaction was complete, the reaction solution was slowly poured into 1.0L ice-water mixture to quench, and extracted with 2×500mL dichloromethane. The organic layers were combined and concentrated. The solid was washed with methanol (500 mL×2) to obtain compound 1b as a yellow solid (48.4 g, yield 66%).

LC-MS m/z (ESI) = 375.00 [M+1].

Step two:

4-Bromo-5-methoxy-2-(4-methoxybenzyl)pyridazin-3(2H)-one (1c)

4-bromo-5-methoxy-2-(4-methoxybenzyl)pyridazin-3(2H)-one

Compound 1b (48.4g, 129.40mmol, 1.0equiv), potassium hydroxide (21.78g, 388.30mmol, 3.00equiv) was dissolved in methanol (417mL), and the reaction solution was stirred at room temperature for 2h. The resulting reaction mixture was concentrated to 80 mL and filtered to obtain a crude product. The obtained filter cake was slurried in water (160 mL) for 1 h and filtered to obtain compound 1c as a white solid (38.72 g, 92% yield).

LC-MS m/z (ESI) = 326.30 [M+1].

third step:

5-methoxy-2-(4-methoxybenzyl)-4-(trifluoromethyl)pyridazin-3(2H)-one (1d)

5-methoxy-2-(4-methoxybenzyl)-4-(trifluoromethyl)pyridazin-3(2H)-one

Weigh compound 1c (14g, 43.04mmol, 1.0equiv) and cuprous iodide (4.10g, 21.52mmol, 0.50equiv) and in a 250mL reaction flask, dissolve with N-methylpyrrolidone (72mL), then slowly add 2, Methyl 2-difluoro-2-(fluorosulfonyl)acetate (16.4 mL, 129.11 mmol, 3.0 equiv). After the addition was complete, the reaction was placed in an oil bath at 100°C and stirred for 3h. After the reaction was complete, 90 mL of water was added to the reaction solution to quench it. The resulting solution was extracted with 3 x 60 mL of dichloromethane. The organic layers were combined, dried over anhydrous sodium sulfate, concentrated in vacuo, and the residue was purified by column chromatography (petroleum ether: ethyl acetate = 1:1) to obtain compound 1d as a white solid (12.1 g, yield 89%).

LC-MS m/z (ESI) = 315.10 [M+1].

the fourth step:

5-Hydroxy-2-(4-methoxybenzyl)-4-(trifluoromethyl)pyridazin-3(2H)-one (1e)

5-hydroxy-2-(4-methoxybenzyl)-4-(trifluoromethyl)pyridazin-3(2H)-one

To a solution of compound 1d (12.1 g, 38.52 mmol, 1.0 equiv) in N,N-dimethylformamide (60 mL) was added dropwise iodotrimethylsilane (9.97 g, 50.07 mmol, 1.3 equiv ). The resulting reaction solution was stirred at 85 °C for 20 h. After the reaction was completed, 60 mL of water was added to the reaction mixture to quench the reaction, and then the resulting solution was extracted with 3×60 mL of dichloromethane. The organic phases were combined, dried over anhydrous sodium sulfate, concentrated in vacuo, and the residue was purified by column chromatography (petroleum ether: ethyl acetate = 1:1) to obtain compound 1e as a white solid (10.4 g, yield 90%).

LC-MS m/z (ESI) = 301.07 [M+1].

the fifth step:

5-Chloro-2-(4-methoxybenzyl)-4-(trifluoromethyl)pyridazin-3(2H)-one (1f)

5-chloro-2-(4-methoxybenzyl)-4-(trifluoromethyl)pyridazin-3(2H)-one

To a solution of compound 1e (10.4 g, 34.66 mmol, 1.0 equiv) in DMF (52 mL) was slowly added dropwise oxalyl dichloride (8.79 g, 69.32 mmol, 2.0 equiv) at 0°C. After the addition was complete, the reaction mixture was stirred at room temperature for 8 h. After the reaction was complete, 550 mL of water was added to the reaction solution to quench it. The mixture was filtered to obtain compound 1f as a white solid (11.04 g, 99%).

LC-MS m/z (ESI) = 319.68 [M+1].

Step six:

(S)-5-((1-hydroxypropan-2-yl)amino)-2-(4-methoxybenzyl)-4-(trifluoromethyl)pyridazin-3(2H)-one ( 1g)

(S)-5-((1-hydroxypropan-2-yl)amino)-2-(4-methoxybenzyl)-4-(trifluoromethyl)pyridazin-3(2H)-one

Weigh compound 1f (5.0 g, 15.69 mmol, 1.0 equiv) into a 100 mL reaction flask, and add ethanol (40 mL) to dissolve it. Subsequently, triethylamine (4.4mL, 31.38mmol, 2.0equiv) and (2S)-2-aminopropan-1-ol (1.19g, 15.69mmol, 1.0equiv) were added successively, and the mixture was stirred at 60°C for 4h . After the reaction was complete, it was concentrated in vacuo, and the residue was purified by column chromatography (petroleum ether:ethyl acetate=1:1.5) to obtain compound 1g as a white solid (5.31g, yield 95%).

LC-MS m/z (ESI) = 358.43 [M+1].

Step seven:

(S)-3-(2-(((1-(4-methoxybenzyl)-6-oxo-5-(trifluoromethyl)-1,6-dihydropyridazin-4-yl )amino)propoxy)methyl propionate (1h)

Methyl-(S)-3-(2-((1-(4-methoxybenzyl)-6-oxo-5-(trifluoromethyl)-1,6-dihydropyridazin-4-yl)amino)propoxy)propanoate

Weigh compound 1g (5.31g, 14.86mmol, 1.0equiv), cesium carbonate (4.73g, 14.86mmol, 1.0equiv) in a 100mL reaction flask. Subsequently, acetonitrile (50 mL) was added for dissolution, and methyl prop-2-enoate (13.38 g, 148.6 mmol, 10 equiv) was added to the mixture. After the addition was complete, the reaction mixture was stirred at 30 °C for 4 h. After the reaction was complete, it was concentrated in vacuo, and the residue was purified by column chromatography (petroleum ether:ethyl acetate=1:1) to obtain compound 1h as a white solid (3.40 g, yield 51%).

LC-MS m/z (ESI) = 444.50 [M+1].

Step eight:

(S)-methyl 3-(2-((6-oxo-5-(trifluoromethyl)-1,6-dihydropyridazin-4-yl)amino)propoxy)propanoate (1i )

Methyl-(S)-3-(2-((6-oxo-5-(trifluoromethyl)-1,6-dihydropyridazin-4-yl)amino)propoxy)propanoate

To a 25 mL reaction flask weighing compound 1h (3.40 g, 7.67 mmol, 1.0 equiv) was added trifluoroacetic acid (21.42 mL) and trifluoromethanesulfonic acid (2.7 mL, 30.68 mmol, 4.0 equiv) sequentially. After the addition was complete, the reaction was stirred at 25 °C for 2 h. Subsequently, 85 mL of water was added to the reaction solution to quench it. The resulting solution was extracted with 3 x 60 mL ethyl acetate. The pH of the organic layer was adjusted to 8-9 by aqueous potassium carbonate. The organic layers were combined and concentrated in vacuo, and the residue was purified by column chromatography (petroleum ether:ethyl acetate=1:3) to obtain compound 1i as a white solid (2.06 g, yield 83%).

LC-MS m/z (ESI) = 324.30 [M+1].

Step Nine:

(S)-3-(2-((6-Oxo-5-(trifluoromethyl)-1,6-dihydropyridazin-4-yl)amino)propoxy)propanoic acid (Intermediate 1 )

(S)-3-(2-((6-oxo-5-(trifluoromethyl)-1,6-dihydropyridazin-4-yl)amino)propoxy)propanoic acid

Compound 1i (2.06g, 6.37mmol, 1.0equiv) and lithium hydroxide monohydrate (1.34g, 31.85mmol, 5.0equiv) were weighed and added to a 100mL reaction flask. Subsequently, methanol (14 mL) and water (14 mL) were added sequentially, and the reaction solution was stirred at 25° C. for 1 h. After the reaction was complete, the pH value of the reaction mixture was adjusted to 6 with trifluoroacetic acid. The reaction mixture was concentrated in vacuo, and the residue was purified by C18 reverse phase chromatography (water:acetonitrile=6:1) to obtain Intermediate 1 as a pale yellow solid (1.32 g, yield 67%).

LC-MS m/z (ESI) = 310.32 [M+1].

Intermediate 2

5-Cyclopropyl-2-(piperazin-1-yl)pyrimidine (Intermediate 2)

5-cyclopropyl-2-(piperazin-1-yl)pyrimidine

first step:

tert-butyl 4-(5-bromopyrimidin-2-yl)piperazine-1-carboxylate (2b)

tert-butyl 4-(5-bromopyrimidin-2-yl)piperazine-1-carboxylate

Weigh 2-chloro-5-bromopyrimidine (2a, 12.0g, 62.04mmol, 1.05equiv), piperazine-1-carboxylate tert-butyl ester (11.0g, 59.08mmol, 1.0equiv) and potassium carbonate (16.33g, 124.08mmol, 2.0equiv) in a 250mL reaction flask. Subsequently, N-methylpyrrolidone (90 mL) was added to the mixture to dissolve, and the reaction was heated and stirred in an oil bath at 80° C. for 1 h. After the reaction was complete, the reaction solution was poured into 280 mL of ice-water mixture, and the suspension was filtered to obtain compound 2b as a white solid (16.4 g, yield 81%).

LC-MS m/z (ESI) = 344.90 [M+1].

Step two:

5-Bromo-2-(piperazin-1-yl)pyrimidine (2c)

5-bromo-2-(piperazin-1-yl)pyrimidine

Compound 2b (16.4 g, 47.78 mmol, 1.0 equiv) was weighed and dissolved in a solution of hydrochloric acid in dioxane (4M, 67 mL). The reaction was stirred at room temperature for 1 h. When the reaction was complete, the suspension was filtered, the filter cake was washed with dioxane, and dried to obtain compound 2c as a white solid (11.3 g, yield 97%).

LC-MS m/z (ESI) = 244.90 [M+1].

third step:

5-Cyclopropyl-2-(piperazin-1-yl)pyrimidine (Intermediate 2)

5-cyclopropyl-2-(piperazin-1-yl)pyrimidine

In a 25mL reaction flask, add compound 2c (1.0g, 4.10mmol, 1.0equiv), potassium carbonate (2.83g, 20.50mmol, 5.0equiv), cyclopropaneboronic acid (1.06g, 12.30mmol, 3.0equiv) respectively ) and [1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium dichloromethane complex (344.8 mg, 0.41 mmol, 0.1 equiv). Then THF/H ₂ O (35.2/4.4 mL) mixture was added, under N ₂ protection, and the reaction was heated and stirred in an oil bath at 66° C. for 3 h. After the reaction was complete, the reaction solution was cooled to room temperature, concentrated in vacuo, and purified by column chromatography (dichloromethane:methanol=15:1) to obtain intermediate 2 as a white solid (0.66 g, yield 78%).

LC-MS m/z (ESI) = 205.30 [M+1].

Intermediate 3

5-Cyclobutyl-2-(piperazin-1-yl)pyrimidine (Intermediate 3)

5-cyclobutyl-2-(piperazin-1-yl)pyrimidine

first step:

tert-butyl 4-(5-(1-(hydroxycyclobutyl)pyrimidin-2-yl)piperazine-1-carboxylate (3a)

tert-butyl 4-(5-(1-hydroxycyclobutyl)pyrimidin-2-yl)piperazine-1-carboxylate

Compound 2b (7.8 g, 22.66 mmol, 1.0 equiv) was dissolved in anhydrous THF (62 mL), and cooled to -78°C. Subsequently, 23.8 mL of n-butyl lithium (1.0 M n-hexane solution, 23.79 mmol, 1.05 equiv) was added dropwise. After the addition was completed, after stirring at this temperature for 10 min, the temperature was naturally raised to ambient temperature and stirring was continued for 2 h. The reaction solution was cooled to -20°C, and cyclobutanone (1.86 mL, 24.93 mmol, 1.1 equiv) was slowly added dropwise. After the addition was complete, the reaction mixture was allowed to warm to ambient temperature and stirred for 4 h. After the reaction was complete, the reaction was quenched with 10 mL of saturated ammonium chloride solution. The reaction mixture was extracted with 3 x 120 mL of ethyl acetate. The organic layers were combined, dried over anhydrous sodium sulfate, and concentrated in vacuo to obtain crude product 3a, 6.4 g of yellow oil. The crude product 3a was used directly in the next reaction without further purification.

LC-MS m/z (ESI) = 335.40 [M+1].

Step two:

5-Cyclobutyl-2-(piperazin-1-yl)pyrimidine (Intermediate 3)

5-cyclobutyl-2-(piperazin-1-yl)pyrimidine

The dried compound 3a (about 5.7 mmol) was transferred to a 20 mL reaction flask, and dichloromethane (6 mL) was dissolved. Subsequently, triethylsilane (2.74 mL, 17.1 mmol, 3 equiv) and a dichloromethane solution of trifluoroacetic acid (3.0 mL, TFA:CH ₂ Cl ₂ =1:2, volume ratio) were added thereto. After the addition was complete, the reaction mixture was stirred overnight at room temperature. The reaction solution was concentrated in vacuo to obtain an oil. The residue was purified by reverse-phase C18 column chromatography (water:acetonitrile=1:3) to obtain Intermediate 3 as a white solid (1.35 g, 27%).

LC-MS m/z (ESI) = 219.20 [M+1].

Intermediate 4

5-(1-fluorocyclopropyl)-2-(piperazin-1-yl)pyrimidine (intermediate 4)

5-(1-fluorocyclopropyl)-2-(piperazin-1-yl)pyrimidine

first step:

Methyl 2-(4-(tert-butoxycarbonyl)piperazin-1-yl)pyrimidine-5-carboxylate (4b)

methyl 2-(4-(tert-butoxycarbonyl)piperazin-1-yl)pyrimidine-5-carboxylate

Weigh 2-chloro-5-carboxylic acid methyl ester 4a (17.2g, 0.1mol, 1.0equiv), piperazine-1-carboxylic acid tert-butyl ester (21.0g, 0.11mol, 1.1equiv) and potassium carbonate (28.0g , 0.2mol, 2.0equiv) in a 250mL reaction flask. Subsequently, N-methylpyrrolidone (100 mL) was added to the mixture to dissolve, and the reaction was heated and stirred in an oil bath at 80 °C for 1 h. After the reaction was complete, the reaction solution was poured into 280 mL of ice-water mixture, and the suspension was filtered to obtain compound 4b as a white solid (26.2 g, yield 81%).

¹ H NMR (400 MHz, DMSO-d ₆ ) δ 8.81 (s, 2H), 3.92-3.69 (m, 7H), 3.47-3.37 (m, 4H), 1.42 (s, 9H).

LC-MS m/z (ESI) = 323.20 [M+1].

Step two:

tert-butyl 4-(5-(1-(hydroxycyclopropyl)pyrimidin-2-yl)piperazine-1-carboxylate (4c)

tert-butyl 4-(5-(1-hydroxycyclopropyl)pyrimidin-2-yl)piperazine-1-carboxylate

Weigh compound 4b (5.0g, 15.5mmol, 1.0equiv), tetraisopropyl titanate (6.4g, 21.7mmol, 1.4equiv) dissolved in 50mL of anhydrous tetrahydrofuran, add ethyl bromide dropwise in an ice-water bath under nitrogen protection Magnesium chloride solution (43.5 mL, 43.5 mol, 2.8 equiv) was added dropwise and reacted at room temperature for 2 h. After the reaction was complete, 40 mL of ethyl acetate and 20 mL of water were added to the reaction liquid, extracted three times with ethyl acetate, concentrated under reduced pressure to remove the solvent, and the crude product was subjected to column chromatography (n-hexane:ethyl acetate=5:1) to obtain compound 4c , as a white solid (1.5 g, 30% yield).

LC-MS m/z (ESI) = 321.20 [M+1].

third step:

tert-butyl 4-(5-(1-(((trimethylsilyl)oxy)cyclopropyl)pyrimidin-2-yl)piperazine-1-carboxylate (4d)

tert-butyl-4-(5-(1-((trimethylsilyl)oxy)cyclopropyl)pyrimidin-2-yl)piperazine-1-carboxylate

Weigh compound 4c (500 mg, 1.5 mmol, 1.0 equiv) and dissolve it in 5 mL of dimethylformamide, add imidazole (204 mg, 3 mol, 2 equiv) and trimethylchlorosilane (285 μL, 2.25 mmol, 1.5 equiv) successively, and react at room temperature 1h. After the reaction was complete, it was quenched by adding 5 mL of water, extracted with ethyl acetate, and the organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure to obtain compound 4d, which was directly used in the next reaction.

the fourth step:

tert-butyl 4-(5-(1-fluorocyclopropyl)pyrimidin-2-yl)piperazine-1-carboxylate (4e)

tert-butyl 4-(5-(1-fluorocyclopropyl)pyrimidin-2-yl)piperazine-1-carboxylate

Compound 4d (1.5 mmol, 1.0 equiv) was dissolved in 5 mL of dichloromethane, diethylaminosulfur trifluoride (396 μL, 3 mol, 2.0 equiv) was added, and reacted at room temperature for 1 h. After the reaction was complete, it was directly spin-dried and passed through the column (n-hexane:ethyl acetate=3:1) to obtain compound 4e as a white solid (290 mg, yield 60%).

¹ H NMR (400MHz,DMSO-d ₆ )δ8.44(d,2H),3.77–3.71(m,4H),3.40(dd,4H),1.42(s,9H),1.39–1.31(m,2H ), 1.12–1.01(m,2H).

the fifth step:

5-(1-fluorocyclopropyl)-2-(piperazin-1-yl)pyrimidine (intermediate 4)

5-(1-fluorocyclopropyl)-2-(piperazin-1-yl)pyrimidine

Referring to the synthesis method of compound 2c, intermediate 4 was prepared as a white solid, and intermediate 4 could be directly used in the next reaction without further purification.

LC-MS m/z(ESI)=223.20[M+1]

Intermediate 5

5-Cyclopropylpyrimidine-2-((3S,5R)-3,5-dimethylpiperazin-1-yl)pyrimidine (Intermediate 5)

5-cyclopropyl-2-((3S,5R)-3,5-dimethylpiperazin-1-yl)pyrimidine

first step:

(2S,6R)-4-(5-Bromopyrimidin-2-yl)-2,6-dimethylpiperazine-1-carboxylic acid tert-butyl ester (5b)

tert-butyl(2S,6R)-4-(5-bromopyrimidin-2-yl)-2,6-dimethylpiperazine-1-carboxylate

Weigh 2-chloro-5-bromopyrimidine 2a (1.5g, 7.7mmol, 1.0equiv), (2S, 6R)-2,6-dimethylpiperazine-1-carboxylic acid tert-butyl ester (2.0g, 9.0mmol , 1.2equiv) and potassium carbonate (2.1g, 15.4mmol, 2.0equiv) in a 100mL reaction flask. Subsequently, nitrogen methylpyrrolidone (20 mL) was added to the mixture to dissolve, and the reaction was heated and stirred in an oil bath at 80° C. for 1 h. After the reaction was complete, the reaction solution was poured into 80 mL of ice-water mixture, and the suspension was filtered to obtain compound 5b as a white solid (2.7 g, yield 98%).

LC-MS m/z (ESI) = 371.20 [M+1].

Step two:

(2S,6R)-4-(5-cyclopropylpyrimidin-2-yl)-2,6-dimethylpiperazine-1-carboxylic acid tert-butyl ester (5c)

tert-butyl(2S,6R)-4-(5-cyclopropylpyrimidin-2-yl)-2,6-dimethylpiperazine-1-carboxy-late

Referring to the synthesis method of Intermediate 2, Compound 5c was prepared as a white solid (1.8 g, yield 70%).

¹ H NMR (400MHz,DMSO-d ₆ )δ8.17(s,2H),4.52(s,1H),4.49(s,1H),4.19–4.08(m,2H),2.99(dd,2H), 1.76–1.64(m,1H), 1.42(s,9H), 1.09(d,6H), 0.89–0.83(m,2H), 0.67–0.59(m,2H).

LC-MS m/z (ESI) = 333.23 [M+1].

third step:

5-Cyclopropylpyrimidine-2-((3S,5R)-3,5-dimethylpiperazin-1-yl)pyrimidine (Intermediate 5)

5-cyclopropyl-2-((3S,5R)-3,5-dimethylpiperazin-1-yl)pyrimidine

Referring to the synthesis method of compound 2c, intermediate 5 was prepared as a white solid, and intermediate 5 was directly used in the next reaction without further purification.

Intermediate 6

1-(5-Cyclopropylpyrimidin-2-yl)-3,3-difluoropiperidin-4-amine (Intermediate 6)

1-(5-cyclopropylpyrimidin-2-yl)-3,3-difluoropiperidin-4-amine

first step:

(1-(5-Bromopyrimidin-2-yl)-3,3-difluoropiperidin-4-yl)carbamate tert-butyl ester (6b)

tert-butyl(1-(5-bromopyrimidin-2-yl)-3,3-difluoropiperidin-4-yl)carbamate

Weigh 2-chloro-5-bromopyrimidine 2a (682 mg, 3.5 mmol, 1.0 equiv), (3,3-difluoropiperidin-4-yl) tert-butyl carbamate (1.0 g, 4.2 mmol, 1.2 equiv) and potassium carbonate (0.97g, 7.0mmol, 2.0equiv) in a 50mL reaction flask. Subsequently, nitrogen methylpyrrolidone (10 mL) was added to the mixture to dissolve, and the reaction was heated and stirred in an oil bath at 80° C. for 1 h. After the reaction was complete, the reaction solution was poured into 80 mL of ice-water mixture, and the suspension was filtered to obtain compound 6b as a white solid (1.02 g, yield 75%).

¹ H NMR (400MHz,DMSO-d ₆ )δ8.50(s,2H),7.24(d,1H),4.76–4.68(m,1H),4.45(d,1H),4.23–4.04(m,1H ), 3.56(dd,1H), 3.30–3.23(m,1H), 1.83–1.82(m,1H), 1.67–1.54(m,1H), 1.40(s,9H).

LC-MS m/z (ESI) = 394.25 [M+1].

Step two:

(1-(5-Cyclopropylpyrimidin-2-yl)-3,3-difluoropiperidin-4-yl)carbamate tert-butyl ester (6c)

tert-butyl(1-(5-cyclopropylpyrimidin-2-yl)-3,3-difluoropiperidin-4-yl)carbamate

Referring to the synthesis method of Intermediate 2, Intermediate 6c was prepared as a white solid (770 mg, yield 87%).

¹ H NMR (600MHz,DMSO-d ₆ )δ8.19(s,2H),7.20(d,1H),4.78–4.72(m,1H),4.47(d,1H),4.16–4.02(m,1H ),3.50–3.43(m,1H),3.21–3.16(m,1H),1.79–1.74(m,2H),1.62–1.56(m,1H),1.40(s,9H),0.93–0.83(m ,2H),0.68–0.61(m,2H).

LC-MS m/z (ESI) = 355.40 [M+1].

third step:

1-(5-Cyclopropylpyrimidin-2-yl)-3,3-difluoropiperidin-4-amine (Intermediate 6)

1-(5-cyclopropylpyrimidin-2-yl)-3,3-difluoropiperidin-4-amine

Referring to the synthesis method of compound 2c, intermediate 6 was prepared as a white solid, and intermediate 6 was directly used in the next reaction without further purification.

Intermediate 7:

7-(5-Cyclopropylpyrimidin-2-yl)-4,7-diazaspiro[2.5]octane (Intermediate 7)

7-(5-cyclopropylpyrimidin-2-yl)-4,7-diazaspiro[2.5]octane

first step:

2-Chloro-5-cyclopropylpyrimidine (7a)

2-chloro-5-cyclopropylpyrimidine

Referring to the synthesis method of Intermediate 2, Compound 7a was prepared as a white solid (1.32 g, yield 83%).

¹ H NMR (400 MHz, DMSO-d ₆ ) δ 8.55 (s, 2H), 2.00–1.93 (m, 1H), 1.09–1.02 (m, 2H), 0.90–0.84 (m, 2H).

LC-MS m/z (ESI) = 155.03 [M+1].

Step two:

tert-butyl 7-(5-cyclopropylpyrimidin-2-yl)-4,7-diazaspiro[2.5]octane-4-carboxylate (7b)

tert-butyl 7-(5-cyclopropylpyrimidin-2-yl)-4,7-diazaspiro[2.5]octane-4-carboxylate

Weigh 2-chloro-5-cyclopropylpyrimidine 7a (300mg, 2mmol, 1.0equiv), 4,7-diazaspiro[2.5]octane-4-carboxylic acid tert-butyl ester (640mg, 3mmol, 1.2equiv ) and potassium carbonate (0.55g, 4.0mmol, 2.0equiv) in a 50mL reaction flask. Subsequently, nitrogen methylpyrrolidone (10 mL) was added to the mixture to dissolve, and the reaction was heated and stirred in an oil bath at 80° C. for 1 h. After the reaction was complete, the reaction solution was poured into 80 mL of ice-water mixture, and the suspension was filtered to obtain compound 7b as a white solid (350 mg, yield 53%).

LC-MS m/z (ESI) = 331.30 [M+1].

third step:

Referring to the synthesis method of compound 2c, intermediate 7 was prepared as a white solid; intermediate 7 could be directly used in the next reaction without further purification.

Intermediate 8:

(1R,5S)-3-(5-Cyclopropylpyrimidin-2-yl)-1,5-dimethyl-3-azabicyclo[3.1.0]hexan-6-amine (Intermediate 8)

(1R,5S)-3-(5-cyclopropylpyrimidin-2-yl)-1,5-dimethyl-3-azabicyclo[3.1.0]hexan-6-amine

Referring to the synthesis method of Intermediate 7, Intermediate 8 was prepared as a white solid (1.8 g, yield 70%).

LC-MS m/z(ESI)=344.50[M+1]

Intermediate 9:

(S)-2-(2-(((1-(4-methoxybenzyl)-6-oxo-5-(trifluoromethyl)-1,6-dihydropyridazin-4-yl )amino)propoxy)acetic acid (Intermediate 9)

(S)-2-(2-((1-(4-methoxybenzyl)-6-oxo-5-(trifluoromethyl)-1,6-dihydropyridazin-4-yl)amino)propoxy)acetic acid

first step:

(S)-2-(2-(((1-(4-methoxybenzyl)-6-oxo-5-(trifluoromethyl)-1,6-dihydropyridazin-4-yl ) amino) propoxy) tert-butyl acetate (9a)

tert-butyl(S)-2-(2-((1-(4-methoxybenzyl)-6-oxo-5-(trifluoromethyl)-1,6-dihydro pyridazin-4-yl)amino)propoxy)acetate

Weigh compound 1g (1.0g, 2.79mmol, 1.0equiv), tert-butyl bromoacetate (1.65g, 8.37mmol, 3.0equiv) and potassium tert-butoxide (0.94g, 8.37mmol, 3.0equiv) in 11.0mL di In methylformamide, react at room temperature for 3 h, add 30 mL of water to quench the reaction, then extract twice with 30 mL of ethyl acetate, dry the organic phase with anhydrous sodium sulfate, concentrate, and column chromatography of the residue (petroleum ether: ethyl acetate =2:1) Compound 9a was obtained as a yellow solid (0.33 g, yield 25%).

LC-MS m/z (ESI) = 472.48 [M+1].

Step two:

(S)-2-(2-(((1-(4-methoxybenzyl)-6-oxo-5-(trifluoromethyl)-1,6-dihydropyridazin-4-yl )amino)propoxy)acetic acid (Intermediate 9)

(S)-2-(2-((1-(4-methoxybenzyl)-6-oxo-5-(trifluoromethyl)-1,6-dihydropyridazin-4-yl)amino)propoxy)acetic acid

In a 50 mL single-necked bottle, compound 9a (330 mg, 0.7 mmol, 1 equiv) was dissolved in 2.5 mL of dichloromethane, then 2.5 mL of trifluoroacetic acid was added, reacted at room temperature for 2 h, and the reaction solution was spin-dried to obtain intermediate 9 (220 mg, yield Yield 76%), carry out next step reaction directly.

LC-MS m/z (ESI) = 416.37 [M+1].

Intermediate 10

1-(2-(Piperazin-1-yl)pyrimidin-5-yl)cyclopropane-1-carbonitrile (Intermediate 10)

1-(2-(piperazin-1-yl)pyrimidin-5-yl)cyclopropane-1-carbonitrile

first step:

tert-butyl 4-(5-(1-(cyanocyclopropyl)pyrimidin-2-yl)piperazine-1-carboxylate (10a)

tert-butyl 4-(5-(1-cyanocyclopropyl)pyrimidin-2-yl)piperazine-1-carboxylate

To the reaction flask was added 1,1'-binaphthyl-2,2'-bisdiphenylphosphine (124.5 mg, 0.2 mmol, 0.1 equiv), tris(dibenzylideneacetone) dipalladium(0):1,1 '-binaphthyl-2,2'-bisdiphenylphosphine (91.6 mg, 0.1 mmol, 0.05 equiv) and tetrahydrofuran (2.0 ml) were pumped into nitrogen, and the suspension was stirred under nitrogen for 20 min. Take another reaction bottle, add compound 2b (686mg, 2.0mmol, 1.0equiv) and cyclopropanecarbonitrile (267μL, 3.0mmol, 1.5equiv), add cyclopentyl methyl ether (4.0mL) to dissolve, and replace into nitrogen protection. Subsequently, the prepared catalyst suspension was added to the reaction, followed by dropwise addition of lithium bistrimethylsilylamide (3.0 mL, 1 M in THF, 3.0 mmol, 1.5 euqiv). The reaction mixture was heated to 80° C. and stirred for 3 h. After the reaction was complete, the reaction solution was cooled to room temperature and diluted with ethyl acetate and saturated ammonium chloride solution. The reaction mixture was extracted with 3 x 60 mL of ethyl acetate. The organic layers were combined, dried over anhydrous sodium sulfate, concentrated in vacuo, and purified by column chromatography (ethyl acetate:petroleum ether=1:10) to obtain compound 10a as a white solid (482 mg, yield 73%).

LC-MS m/z (ESI) = 330.30 [M+1].

Step two:

1-(2-(Piperazin-1-yl)pyrimidin-5-yl)cyclopropane-1-carbonitrile (Intermediate 10)

1-(2-(piperazin-1-yl)pyrimidin-5-yl)cyclopropane-1-carbonitrile

Referring to the synthesis method of compound 2c, the crude product of intermediate 10 was prepared as 327 mg of white solid. Intermediate 10 can be directly used in the next reaction without further purification.

LC-MS m/z (ESI) = 230.20 [M+1].

Intermediate 11

5-(2,2-Difluorocyclopropyl)-2-(piperazin-1-yl)pyrimidine (Intermediate 11)

5-(2,2-difluorocyclopropyl)-2-(piperazin-1-yl)pyrimidine

first step:

tert-butyl 4-(5-vinylpyrimidin-2-yl)piperazine-1-carboxylate (11b)

tert-butyl 4-(5-vinylpyrimidin-2-yl)piperazine-1-carboxylate

Referring to the synthetic method of compound 2b, compound 11b was prepared as a white solid (1.2 g, yield 83%).

LC-MS m/z (ESI) = 290.4 [M+1].

Step two:

tert-butyl 4-(5-(2,2-difluorocyclopropyl)pyrimidin-2-yl)piperazine-1-carboxylate (11c)

tert-butyl 4-(5-(2,2-difluorocyclopropyl)pyrimidin-2-yl)piperazine-1-carboxylate

Weigh respectively 11b (634mg, 2.18mmol, 1.0eq), sodium iodide (729.7mg, 4.87mmol, 2.23equiv) in a 15mL high-pressure reaction tube, add dioxane (2.0mL) and ethylene glycol dimethyl ether (30 μL) to dissolve. Pumping into nitrogen protection, followed by dropwise addition of methyl fluorosulfonyl difluoroacetate (556μL, 4.37mmol, 2.0equiv) and trimethylchlorosilane (474.3mg, 4.37mmol, 2.0equiv), after addition, the reaction was transferred to Stir the reaction in an oil bath at 120°C for 3h. After the reaction was complete, the reaction solution was cooled to room temperature, 30 mL of ethyl acetate and 30 mL of water were added, and the mixture was stirred for 30 min. The organic phase was separated, concentrated in vacuo, and purified by column chromatography (ethyl acetate:petroleum ether=1:15) to obtain compound 11c as a white solid (220 mg, yield 30%).

LC-MS m/z (ESI) = 341.30 [M+1].

third step:

5-(2,2-Difluorocyclopropyl)-2-(piperazin-1-yl)pyrimidine (Intermediate 11)

5-(2,2-difluorocyclopropyl)-2-(piperazin-1-yl)pyrimidine

Referring to the synthesis method of compound 2c, the crude product of intermediate 11 was prepared as a white solid (169 mg). Intermediate 11 can be directly used in the next reaction without further purification.

LC-MS m/z (ESI) = 241.29 [M+1].

Intermediate 12

2-(Piperazin-1-yl)-5-(2,2,3-trifluorocyclopropyl)pyrimidine (Intermediate 12)

2-(piperazin-1-yl)-5-(2,2,3-trifluorocyclopropyl)pyrimidine

first step:

tert-butyl 4-(5-(2-bromo-1-fluoroethyl)pyrimidin-2-yl)piperazine-1-carboxylate (12a)

tert-butyl 4-(5-(2-bromo-1-fluoroethyl)pyrimidin-2-yl)piperazine-1-carboxylate

Compound 11b (420 mg, 1.4 mmol, 1.0 equiv) was weighed into a 25 mL reaction flask, and 10 mL of dichloromethane was added to dissolve it. The reaction was lowered to 0°C, then a solution of triethylamine in hydrogen trifluoride (338 mg, 2.1 mmol, 1.5 equiv) was added dropwise, and bromosuccinimide (300 mg, 1.68 mmol, 1.2 equiv) was added to the reaction solution. The reaction was allowed to rise to room temperature naturally, and stirring was continued for 1 h. After the reaction was complete, pour the reaction into 20 mL of ice water, adjust the aqueous phase to be alkaline with concentrated ammonia, extract with dichloromethane (3×40 mL), separate the organic phase, concentrate in vacuo, and dry to obtain compound 12a as 344 mg of light yellow oil. 12a was directly used in the next reaction without further purification.

LC-MS m/z (ESI) = 390.20 [M+1].

Step two:

tert-butyl 4-(5-(1-fluorovinyl)pyrimidin-2-yl)piperazine-1-carboxylate (12b)

tert-butyl 4-(5-(1-fluorovinyl)pyrimidin-2-yl)piperazine-1-carboxylate

Weigh compound 12a (344mg, 0.88mmol, 1.0equiv) and potassium tert-butoxide (188mg, 1.68mmol, 1.2equiv) respectively in a 25mL reaction flask, add tetrahydrofuran (10mL) to dissolve, then stir the reaction at 50°C . After the reaction was complete, the reaction solution was naturally cooled to room temperature, then slowly added to 30 mL of ice-water mixture, dichloromethane (3 × 40 mL) was extracted, the organic phase was separated, concentrated in vacuo, and column chromatography (ethyl acetate:petroleum ether = 1:20) to obtain compound 12b as a white solid (128 mg, yield 47%).

¹ H NMR (400MHz,DMSO-d ₆ )δ8.62(s,2H),5.25(dd,1H),4.82(dd,1H),3.87–3.72(m,4H),3.41(t,4H), 1.42(s,9H).

LC-MS m/z (ESI) = 309.30 [M+1].

third step:

tert-butyl 4-(5-(2,2,3-trifluorocyclopropyl)pyrimidin-2-yl)piperazine-1-carboxylate (12c)

tert-butyl 4-(5-(2,2,3-trifluorocyclopropyl)pyrimidin-2-yl)piperazine-1-carboxylate

Compound 12b (64 mg, 0.2 mmol, 1.0 equiv) and anhydrous sodium iodide (6 mg, 0.04 mmol, 0.2 eq) were weighed and dissolved in 5 mL of anhydrous tetrahydrofuran. Under nitrogen, (trifluoromethyl)trimethylsilane (74 μL, 0.5 mmol, 2.5 equiv) was added to the reaction. After the addition, the reaction was stirred at 65°C for 2h. After the reaction was complete, the reaction solution was concentrated in vacuo and purified by column chromatography (ethyl acetate:petroleum ether=1:15) to obtain compound 12c as a colorless oil (54 mg, yield 75%).

LC-MS m/z (ESI) = 359.33 [M+1].

the fourth step:

2-(Piperazin-1-yl)-5-(2,2,3-trifluorocyclopropyl)pyrimidine (Intermediate 12)

2-(piperazin-1-yl)-5-(2,2,3-trifluorocyclopropyl)pyrimidine

Referring to the synthesis method of compound 2c, the crude intermediate 12 was prepared as a white solid 47 mg. Intermediate 12 can be directly used in the next reaction without further purification.

LC-MS m/z (ESI) = 259.30 [M+1].

Intermediate 13

3-(5-cyclopropylpyrimidin-2-yl)-3,6-diazabicyclo[3.1.1]heptane (intermediate 13)

3-(5-cyclopropylpyrimidin-2-yl)-3,6-diazabicyclo[3.1.1]heptane

first step:

tert-butyl 3-(5-cyclopropylpyrimidin-2-yl)-3,6-diazacyclo[3.1.1]heptane-6-carboxylate (13a)

tert-butyl 3-(5-cyclopropylpyrimidin-2-yl)-3,6-diazabicyclo[3.1.1]heptane-6-carboxylate

Weigh 2-chloro-5-cyclopropylpyrimidine 7a (300mg, 2mmol, 1.0equiv) and dissolve it in N,N-dimethylformamide (10mL), add 3,6-diazacyclo[3.1.1 ] tert-butyl heptane-6-carboxylate (460 mg, 2.3 mmol, 1.2 equiv) and DIPEA (1.6 mL, 9.7 mmol, 5.0 equiv), and the reaction was heated and stirred in an oil bath at 120° C. for 1 h. The reaction solution was cooled to room temperature and distilled under reduced pressure. Compound 13a was obtained by column chromatography (petroleum ether: ethyl acetate = 3:1) as a yellow oil (400 mg, yield 65%).

¹ H NMR (400MHz,DMSO-d ₆ )δ8.20(s,2H),4.21–4.11(m,2H),4.12–3.96(m,2H),3.44(d,2H),2.56–2.51(m ,1H), 1.78–1.76(m,1H), 1.43(d,1H), 1.25(s,9H), 0.91–0.79(m,2H), 0.69–0.57(m,2H).

LC-MS m/z (ESI) = 317.20 [M+1].

Step two:

3-(5-cyclopropylpyrimidin-2-yl)-3,6-diazabicyclo[3.1.1]heptane (intermediate 13)

3-(5-cyclopropylpyrimidin-2-yl)-3,6-diazabicyclo[3.1.1]heptane

Referring to the synthesis method of compound 2c, intermediate 13 was prepared as a white solid. Intermediate 13 can be directly used in the next reaction without further purification.

LC-MS m/z (ESI) = 217.20 [M+1].

Intermediate 14

8-(5-cyclopropylpyrimidin-2-yl)-3,8-diazacyclo[3.2.1]octane (intermediate 14)

8-(5-cyclopropylpyrimidin-2-yl)-3,8-diazabicyclo[3.2.1]octane

Referring to the synthesis method of intermediate 13, the crude product of intermediate 14 was prepared as a white solid. Intermediate 14 can be directly used in the next reaction without further purification.

LC-MS m/z (ESI) = 231.20 [M+1].

Intermediate 15

5-((2S)-1-(3-(3,8-diazacyclo[3.2.1]octane-8-yl)-3-oxopropoxy)propan-2-yl)amino)- 4-(Trifluoromethyl)pyridazin-3(2H)-one (Intermediate 15)

5-(((2S)-1-(3-(3,8-diazabicyclo[3.2.1]octan-8-yl)-3-oxopropoxy)propan-2-yl)amino)-4-(trifluoromethyl)pyridazin -3(2H)-one

first step:

tert-butyl 8-(3-((S)-2-((6-oxo-5-(trifluoromethyl)-1,6-dihydropyridazin-4-yl)amino)propoxy) Propionyl)-3,8-diazacyclo[3.2.1]octane-3-carboxylate (15b)

tert-butyl8-(3-((S)-2-((6-oxo-5-(trifluoromethyl)-1,6-dihydropyridazin-4-yl)amino)-propoxy)-propanoyl)-3,8-diazabicyclo [3.2.1] octane-3-carboxylate

Weigh 15a (212mg, 1.0mmol, 1.0equiv), intermediate 1 (401mg, 1.3mmol, 1.3equiv), N,N,N',N'-tetramethyl-O-(7-azabenzo Triazol-1-yl)urea hexafluorophosphate (380 mg, 1.0 mmol, 1.0 equiv) was dissolved in a 10 mL reaction flask with N,N-dimethylformamide (4.0 mL). N,N-diisopropylethylamine (0.65 mL, 4.0 mmol, 4 equiv) was added to the mixture, and the reaction was stirred at 25 °C for 1 h. After the reaction was complete, the reaction mixture was concentrated in vacuo. The residue was purified by C18 reverse phase chromatography (water:acetonitrile=1:5) to give 15b as a white solid (307 mg, yield 61%).

LC-MS m/z (ESI) = 505.20 [M+1].

Step two:

5-((2S)-1-(3-(3,8-diazacyclo[3.2.1]octane-8-yl)-3-oxopropoxy)propan-2-yl)amino)- 4-(Trifluoromethyl)pyridazin-3(2H)-one (Intermediate 15)

5-(((2S)-1-(3-(3,8-diazabicyclo[3.2.1]octan-8-yl)-3-oxopropoxy)propan-2-yl)amino)-4-(trifluoromethyl)pyridazin -3(2H)-one

Referring to the synthesis method of compound 2c, the crude intermediate 15 was prepared as a white solid 100 mg. Intermediate 15 can be directly used in the next reaction without further purification.

LC-MS m/z (ESI) = 405.20 [M+1].

Intermediate 16

5-Cyclopropyl-2-((2S,5R)-2,5-dimethylpiperazin-1-yl)pyrimidine (Intermediate 16)

5-cyclopropyl-2-((2S,5R)-2,5-dimethylpiperazin-1-yl)pyrimidine

Referring to the synthesis method of intermediate 13, the crude product of intermediate 16 was prepared as a white solid. Intermediate 16 can be directly used in the next reaction without further purification.

LC-MS m/z (ESI) = 233.20 [M+1].

Intermediate 17

5-Cyclopropyl-2-((2S,5R)-2,5-dimethylpiperazin-1-yl)pyrimidine (Intermediate 17)

5-cyclopropyl-2-((2S,5R)-2,5-dimethylpiperazin-1-yl)pyrimidine

Referring to the synthesis method of intermediate 13, the crude product of intermediate 17 was prepared as a white solid. Intermediate 17 can be directly used in the next reaction without further purification.

LC-MS m/z (ESI) = 217.20 [M+1].

Intermediate 18:

2-((S)-2-((1-(4-methoxybenzyl)-6-oxo-5-(trifluoromethyl)-1,6-dihydropyridazin-4-yl) Amino)propoxy)propionic acid (Intermediate 18)

2-((S)-2-((1-(4-methoxybenzyl)-6-oxo-5-(trifluoromethyl)-1,6-dihydropyridazin-4-yl)amino)propoxy)propanoic acid

first step:

tert-butyl 2-((S)-2-((1-(4-methoxybenzyl)-6-oxo-5-(trifluoromethyl)-1,6-dihydropyridazine-4 -yl)amino)propoxy)propionate (18a)

tert-butyl 2-((S)-2-((1-(4-methoxybenzyl)-6-oxo-5-(trifluoromethyl)-1,6-dihydropyridazin-4-yl)amino)propoxy)propanoate

Weigh compound 1g (550mg, 1.539mmol, 1.0equiv), tert-butyl 2-bromopropionate (479mg, 2.309mmol, 1.5equiv) and potassium tert-butoxide (259mg, 2.309mmol, 1.5equiv) in 11.0mL DMF , react at room temperature for 3 h, add 30 mL of water to quench the reaction, then extract twice with 30 mL of ethyl acetate, dry the organic phase with anhydrous sodium sulfate, concentrate, and column chromatography of the residue (petroleum ether: ethyl acetate=3:1 ) afforded compound 18a as a yellow solid (360 mg, 48% yield).

LC-MS m/z (ESI) = 486.5 [M+1].

Step two:

2-((S)-2-((1-(4-methoxybenzyl)-6-oxo-5-(trifluoromethyl)-1,6-dihydropyridazin-4-yl) Amino)propoxy)propionic acid (Intermediate 18)

2-((S)-2-((1-(4-methoxybenzyl)-6-oxo-5-(trifluoromethyl)-1,6-dihydropyridazin-4-yl)amino)propoxy)propanoic acid

In a 20 mL single-necked bottle, compound 18a (300 mg, 0.618 mmol, 1 equiv) was dissolved in 5 mL of DCM, then 2 mL of trifluoroacetic acid was added, and reacted at room temperature for 2 h. After the reaction was complete, the reaction mixture was concentrated in vacuo. The residue was purified by C18 reverse phase chromatography (water:acetonitrile=3:2) to obtain intermediate 18 as a white solid (250 mg, yield 94%).

LC-MS m/z (ESI) = 430.4 [M+1].

Intermediate 19

3-((1-(1-(1-(4-methoxybenzyl)-6-oxo-5-(trifluoromethyl)-1,6-dihydropyridazin-4-yl)pyrrole Alk-2--2-yl)methoxy)propanoic acid (Intermediate 19)

3-((1-(1-(4-methoxybenzyl)-6-oxo-5-(trifluoromethyl)-1,6-dihydropyridazin-4-yl)pyrrolidin-2-yl)methoxy)propanoic acid

first step:

5-(2-(Hydroxymethyl)pyrrolidin-1-yl)-2-(4-methoxybenzyl)-4-(trifluoromethyl)pyridazin-3(2H)-one (19a)

5-(2-(hydroxymethyl)pyrrolidin-1-yl)-2-(4-methoxybenzyl)-4-(trifluoromethyl)pyridazin-3(2H)-one

Weigh 1f (1.5g, 4.72mmol, 1.0equiv) into a 100mL reaction flask, add N,N-dimethylformamide (15mL) to dissolve. Subsequently, diisopropylethylamine (3.3 mL, 18.87 mmol, 4.0 equiv) and pyrrol-2-ylmethanol (0.53 g, 5.19 mmol, 1.1 equiv) were added sequentially. The mixture was stirred at 100 °C for 2 h. After the reaction was complete, it was concentrated in vacuo, and the residue was purified by column chromatography (petroleum ether: ethyl acetate = 2:1) to obtain compound 19a as a white solid (1.59 g, yield 88%).

LC-MS m/z (ESI) = 384.15 [M+1].

Step two:

Pyrrolidin-2-ylmethanolmethyl 3-((1-(1-(4-methoxybenzyl)-6-oxo-5-(trifluoromethyl)-1,6-dihydropyridazine -4-yl)pyrrolidin-2-yl)methoxy)propionate (19b)

Methyl 3-((1-(1-(4-methoxybenzyl)-6-oxo-5-(trifluoromethyl)-1,6-dihydropyridazin-4-yl)pyrrolidin-2-yl )methoxy)propanoate

Weigh compound 19a (1.59g, 4.16mmol, 1.0equiv), cesium carbonate (1.35g, 4.16mmol, 1.0equiv) in a 50mL reaction flask, add acetonitrile (14mL) to dissolve. Methyl prop-2-enoate (3.6 g, 41.55 mmol, 10.0 equiv) was slowly added dropwise to the mixture. After the addition was complete, the reaction mixture was stirred at 30 °C for 4 h. After the reaction was complete, it was concentrated in vacuo, and the residue was purified by column chromatography (petroleum ether: ethyl acetate = 1:1) to obtain compound 19b as a white solid (1.04 g, yield 53%).

LC-MS m/z (ESI) = 470.18 [M+1].

third step:

3-((1-(1-(4-methoxybenzyl)-6-oxo-5-(trifluoromethyl)-1,6-dihydropyridazin-4-yl)pyrrolidine-2 -yl)methoxy)propionic acid (Intermediate 19)

3-((1-(1-(4-methoxybenzyl)-6-oxo-5-(trifluoromethyl)-1,6-dihydropyridazin-4-yl)pyrrolidin-2-yl)methoxy)propanoic acid

Weigh compound 19b (1.04g, 2.217mmol, 1.0equiv) and lithium hydroxide monohydrate (265mg, 11.09mmol, 5.0equiv), respectively, in a 50mL reaction flask. Subsequently, methanol (5 mL) and water (5 mL) were added sequentially, and the reaction solution was stirred at 25° C. for 1 h. After the reaction was complete, the pH of the reaction mixture was adjusted to 6 with trifluoroacetic acid. The reaction mixture was concentrated in vacuo, and the residue was purified by C18 reverse phase chromatography (water:acetonitrile=5:1) to obtain Intermediate 19 as a pale yellow solid (830 mg, yield 82%).

LC-MS m/z (ESI) = 456.17 [M+1].

Intermediate 20

1-(5-Cyclopropyl-3-(trifluoromethyl)pyridin-2-yl)piperazine (Intermediate 20)

1-(5-cyclopropyl-3-(trifluoromethyl)pyridin-2-yl)piperazine

first step:

tert-butyl 4-(5-bromo-3-(trifluoromethyl)pyridin-2-yl)piperazine-1-carboxylate (20b)

tert-butyl 4-(5-bromo-3-(trifluoromethyl)pyridin-2-yl)piperazine-1-carboxylate

Weigh 20a (3.9g, 15.00mmol, 1.0equiv), piperazine-1-carboxylate tert-butyl ester (5.54g, 22.5mmol, 1.5equiv) and potassium carbonate (7.3g, 52.5mmol, 3.5equiv) in 250mL in the reaction vial. Subsequently, acetonitrile (40 mL) was added to the mixture to dissolve, and the reaction was heated and stirred in an oil bath at 80° C. for 2 h. After the reaction was complete, the reaction solution was cooled to room temperature, concentrated in vacuo, and subjected to column chromatography (petroleum ether: ethyl acetate = 5:1) to obtain intermediate 20b as a white solid (1.4 g, yield 22.67%).

LC-MS m/z (ESI) = 409.06 [M+1].

Step two:

tert-butyl 4-(5-cyclopropyl-3-(trifluoromethyl)pyridin-2-yl)piperazine-1-carboxylate (20c)

tert-butyl-4-(5-cyclopropyl-3-(trifluoromethyl)pyridin-2-yl)piperazine-1-carboxylate

In the 25mL reaction bottle, add the weighed intermediate 20b (1.4g, 3.40mmol, 1.0equiv), cesium carbonate (2.78g, 8.50mmol, 2.5equiv), cyclopropaneboronic acid (442mg, 5.1mmol, 1.5equiv ) and [1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium dichloromethane complex (277 mg, 0.34 mmol, 0.1 equiv). Then Toulune/H ₂ O (20/2 mL) mixture was added, pumped into nitrogen protection, and the reaction was heated and stirred in an oil bath at 95° C. for 2 h. After the reaction was complete, the reaction solution was cooled to room temperature, concentrated in vacuo, and column chromatographed (petroleum ether: ethyl acetate = 5:1) to obtain intermediate 20c as a white solid (1.02 g, yield 79.41%).

LC-MS m/z(ESI)=371.18[M+1]

third step:

1-(5-Cyclopropyl-3-(trifluoromethyl)pyridin-2-yl)piperazine (Intermediate 20)

1-(5-cyclopropyl-3-(trifluoromethyl)pyridin-2-yl)piperazine

Referring to the synthetic method of compound 2c, intermediate 20 was prepared as a white solid (800 mg, yield 97%).

LC-MS m/z (ESI) = 271.18 [M+1].

Intermediate 21

5-Cyclopropyl-2-(piperazin-1-yl)nicotinonitrile (Intermediate 21)

5-cyclopropyl-2-(piperazin-1-yl)nicotinonitrile

first step:

2-Chloro-5-cyclopropylnicotinonitrile (21b)

2-chloro-5-cyclopropylnicotinonitrile

In a 25mL reaction flask, add weighed compound 21a (2.17g, 10mmol, 1.0equiv), cesium carbonate (8.2g, 25mmol, 2.5equiv), cyclopropaneboronic acid (1.72g, 20mmol, 2.0equiv) and [ 1,1'-Bis(diphenylphosphino)ferrocene]dichloropalladium dichloromethane complex (816 mg, 1 mmol, 0.1 equiv). Then Toulune/H ₂ O (20/2 mL) mixture was added, nitrogen protection was replaced by pumping, and the reaction was heated and stirred in an oil bath at 95° C. for 2 h. After the reaction was complete, the reaction solution was cooled to room temperature, concentrated in vacuo, and subjected to column chromatography (petroleum ether: ethyl acetate = 5:1) to obtain intermediate 21b as a white solid (670 mg, yield 38.40%).

LC-MS m/z(ESI)=178.03[M+1]

Step two:

tert-butyl 4-(3-cyano-5-cyclopropylpyridin-2-yl)piperazine-1-carboxylate (21c)

tert-butyl 4-(3-cyano-5-cyclopropylpyridin-2-yl)piperazine-1-carboxylate

Referring to the synthesis method of intermediate 2b, intermediate 21c was purified by column chromatography (petroleum ether: ethyl acetate = 3:1) as a white solid (0.8 g, yield 63.80%).

LC-MS m/z (ESI) = 328.19 [M+1].

third step:

5-Cyclopropyl-2-(piperazin-1-yl)nicotinonitrile (Intermediate 21)

5-cyclopropyl-2-(piperazin-1-yl)nicotinonitrile

Referring to the synthesis method of compound 2c, intermediate 21 was prepared as a white solid. Intermediate 21 can be directly used in the next reaction without further purification.

LC-MS m/z (ESI) = 228.19 [M+1].

Intermediate 22

2-Cyclopropyl-5-(piperazin-1-yl)pyrazine (Intermediate 22)

2-cyclopropyl-5-(piperazin-1-yl)pyrazine

first step:

tert-butyl 4-(5-chloropyrazin-2-yl)piperazine-1-carboxylate (22b)

tert-butyl 4-(5-chloropyrazin-2-yl)piperazine-1-carboxylate

Referring to the synthesis method of intermediate 2b, intermediate 22b was purified by column chromatography (petroleum ether:ethyl acetate=3:1) as a pale yellow solid (2.0 g, yield 50%).

LC-MS m/z (ESI) = 298.12 [M+1].

Step two:

tert-butyl 4-(5-cyclopropylpyrazin-2-yl)piperazine-1-carboxylate (22c)

tert-butyl 4-(5-cyclopropylpyrazin-2-yl)piperazine-1-carboxylate

Referring to the synthesis method of Intermediate 2, it was purified by column chromatography (petroleum ether:ethyl acetate=3:1) to obtain Intermediate 22c as a white solid (0.9 g, yield 44%).

LC-MS m/z(ESI)=304.19[M+1]

third step:

2-Cyclopropyl-5-(piperazin-1-yl)pyrazine (Intermediate 22)

2-cyclopropyl-5-(piperazin-1-yl)pyrazine

Referring to the synthesis method of compound 2c, intermediate 22 was prepared as a white solid. Intermediate 22 can be directly used in the next reaction without further purification.

LC-MS m/z (ESI) = 204.19 [M+1].

Intermediate 23

3-((S)-2-Aminopropoxy)-1-(4-(5-cyclopyrimidin-2-yl)piperazin-1-yl)butanone (Intermediate 23)

3-((S)-2-aminopropoxy)-1-(4-(5-cyclopropylpyrimidin-2-yl)piperazin-1-yl)butan-1-one

first step:

1-(4-(5-cyclopropylpyrimidin-2-yl)piperazin-1-yl)-3-hydroxybutane-1-one (23a)

1-(4-(5-cyclopropylpyrimidin-2-yl)piperazin-1-yl)-3-hydroxybutan-1-one

Weigh intermediate 2 (139mg, 0.50mmol, 1.0equiv), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (144mg, 0.75mmol, 1.5equiv), 1 -Hydroxybenzotriazole (101 mg, 0.75 mmol, 1.5 equiv) was dissolved in a 10 mL reaction flask with N,N-dimethylformamide (2.0 mL). N,N-diisopropylethylamine (331 μL, 2.0 mmol, 4 equiv) and 3-hydroxybutyric acid (57 mg, 0.55 mmol, 1.1 equiv) were sequentially added dropwise to the mixture, and the reaction was stirred at 25° C. for 12 h. After the reaction was complete, the reaction mixture was concentrated in vacuo. The residue was purified by C18 reverse phase chromatography (water:acetonitrile=1:5) to obtain intermediate 23a as a pale yellow oil (135 mg, yield 60%).

LC-MS m/z (ESI) = 290.3 [M+1].

Step two:

((2S)-1-((4-(4-(5-cyclopropylpyrimidin-2-yl)piperazin-1-yl)-4-oxobutan-2-yl)oxy)propan-2 -yl) tert-butyl carbamate (23b)

tert-butyl ((2S)-1-((4-(4-(5-cyclopropylpyrimidin-2-yl)piperazin-1-yl)-4-oxobutan-2-yl)oxy)propan-2-yl)carbamate

Into a 50 mL reaction bottle, weigh intermediate 23a (130 mg, 0.45 mmol, 1.0 equiv) and dissolve in anhydrous N,N-dimethylformamide (4 mL). Under nitrogen protection, sodium hydride (54 mg, 1.35 mmol, 3.0 equiv) was added in batches at 0° C., after the addition was complete, stirring was continued at this temperature for 10 min. Subsequently, (S)-4-methyl-1,2,3-oxathiazolidine-3-carboxylic acid tert-butyl ester 2,2-dioxide (160mg, 0.68mmol, 1.5 equiv) in N,N-dimethylformamide solution (7 mL), keep the dropwise addition process temperature at 0° C. and continue to stir for 2 h. After the reaction was complete, the reaction system was adjusted to pH=3 with hydrochloric acid solution (2M), and stirred at room temperature for 0.5 h. The reaction mixture was extracted with ethyl acetate (3 x 120 mL). The organic layers were combined, dried over anhydrous sodium sulfate, and concentrated in vacuo to obtain a crude product, which was purified by column chromatography (dichloromethane:methanol=20:1) to obtain intermediate 23b as a colorless oil (40 mg, yield 40%) .

LC-MS m/z (ESI) = 448.6 [M+1].

third step:

3-((S)-2-Aminopropoxy)-1-(4-(5-cyclopyrimidin-2-yl)piperazin-1-yl)butanone (Intermediate 23)

3-((S)-2-aminopropoxy)-1-(4-(5-cyclopropylpyrimidin-2-yl)piperazin-1-yl)butan-1-one

Referring to the synthetic method of compound 2c, the crude product of intermediate 23 was prepared as a white solid. Intermediate 23 can be directly used in the next reaction without further purification.

LC-MS m/z (ESI) = 348.5 [M+1].

Intermediate 24

2-(Piperazin-1-yl)-5-(2,2,3,3-tetrafluorocyclopropyl)pyrimidine (Intermediate 24)

2-(piperazin-1-yl)-5-(2,2,3,3-tetrafluorocyclopropyl)pyrimidine

first step:

tert-butyl 4-(5-formylpyrimidin-2-yl)piperazine-1-carboxylate (24c)

tert-butyl 4-(5-formylpyrimidin-2-yl)piperazine-1-carboxylate

Referring to the synthesis method of intermediate 2b, the crude intermediate 24c was obtained as a white solid. Intermediate 24c was directly used in the next reaction without further purification.

LC-MS m/z (ESI) = 293.40 [M+1].

Step two:

tert-butyl 4-(5-(2,2-difluorovinyl)pyrimidin-2-yl)piperazine-1-carboxylate (24d)

tert-butyl 4-(5-(2,2-difluorovinyl)pyrimidin-2-yl)piperazine-1-carboxylate

Weigh intermediate 24c (2.83g, 12mmol, 1.0equiv), triphenylphosphine (4.72g, 18.0mmol, 1.5equiv) in a 250mL reaction flask, dissolve with N,N-dimethylformamide (70mL), Sodium difluorobromoacetate was added in batches under nitrogen protection (completely added within 10 min); after the addition, the reaction was stirred at 90°C for 2 h. After the reaction was complete, the reaction solution was cooled to room temperature and distilled under reduced pressure. The intermediate 24d was obtained as a yellow solid (2.1 g, yield 73%) by column chromatography (petroleum ether: ethyl acetate = 3:1).

LC-MS m/z (ESI) = 327.40 [M+1].

third step:

tert-butyl 4-(5-(2,2,3,3-tetrafluorocyclopropyl)pyrimidin-2-yl)piperazine-1-carboxylate (24e)

tert-butyl 4-(5-(2,2,3,3-tetrafluorocyclopropyl)pyrimidin-2-yl)piperazine-1-carboxylate

Weigh intermediate 24d (1.0 g, 3.1 mmol, 1.0 equiv) into a 100 mL reaction flask, add diethylene glycol dimethyl ether to dissolve, and heat the reaction solution to 180°C under nitrogen protection. When the temperature rises to the specified temperature, sodium chlorodifluoroacetate (1.65 g, 10.8 mmol, 3.5 equiv) is added in batches under nitrogen bubbling; after the addition is complete, the reaction is stirred for 4 h. After the reaction was complete, the residue was purified by C18 reverse phase chromatography (water:acetonitrile=1:5) to obtain intermediate 24e as a white solid (354.4 mg, yield 30%).

LC-MS m/z (ESI) = 377.4 [M+1].

the fourth step:

2-(Piperazin-1-yl)-5-(2,2,3,3-tetrafluorocyclopropyl)pyrimidine (Intermediate 24)

2-(piperazin-1-yl)-5-(2,2,3,3-tetrafluorocyclopropyl)pyrimidine

Referring to the synthesis method of compound 2c, intermediate 24 was prepared as a white solid. Intermediate 24 can be directly used in the next reaction without further purification.

LC-MS m/z (ESI) = 277.4 [M+1].

Example 1

(S)-5-((1-(3-(4-(5-cyclopropylpyrimidin-2-yl)piperazin-1-yl)-3-oxopropoxy)propan-2-yl) Amino)-4-(trifluoromethyl)pyridazin 3(2H)-one (Compound 1)

(S)-5-((1-(3-(4-(5-cyclopropylpyrimidin-2-yl)piperazin-1-yl)-3-oxopropoxy)propan-2-yl)amino)-4-(trifluoromethyl) pyridazin-3(2H)-one

Weigh intermediate 1 (303mg, 0.979mmol, 1.0equiv), intermediate 2 (200mg, 0.979mmol, 1.0equiv), N,N,N',N'-tetramethyl-O-(7-aza Benzotriazol-1-yl)urea hexafluorophosphate (372 mg, 0.979 mmol, 1.0 equiv) was dissolved in N,N-dimethylformamide (4.0 mL) in a 10 mL reaction flask. N,N-diisopropylethylamine (0.65 mL, 3.916 mmol, 4 equiv) was added to the mixture, and the addition was completed, and the reaction was stirred at 25° C. for 1 h. After the reaction was complete, it was concentrated in vacuo, and the residue was purified by C18 reverse phase chromatography (water:acetonitrile=1:5) to obtain compound 1 as a white solid (125 mg, yield 25%).

¹ H NMR (400MHz, Chloroform-d) δ10.33(s,1H),8.21(s,2H),7.66(s,1H),5.78(s,1H),3.97–3.80(m,6H),3.72 –3.71(m,2H),3.64(dd,1H),3.61–3.44(m,3H),2.62(t,2H),1.79–1.73(m,1H),1.31–1.25(m,3H),0.98 –0.95(m,2H),0.66–0.63(m,2H).

LCMS m/z = 496.5 [M+l].

Example 2

(S)-5-((1-(3-(4-(5-cyclobutylpyrimidin-2-yl)piperazin-1-yl)-3-oxopropoxy)propan-2-yl) Amino)-4-(trifluoromethyl)pyridazin 3(2H)-one (compound 2)

(S)-5-((1-(3-(4-(5-cyclobutylpyrimidin-2-yl)piperazin-1-yl)-3-oxopropoxy)propan-2-yl)amino)-4-(trifluoromethyl) pyridazin-3(2H)-one

Referring to the synthetic method of compound 1, compound 2 was prepared as a white solid (157 mg, yield 34%).

¹ H NMR (400MHz, Chloroform-d) δ10.36(s,1H),8.29(s,2H),7.66(s,1H),5.78(d,1H),3.99–3.79(m,6H),3.78 –3.68(m,2H),3.68–3.54(m,3H),3.52–3.35(m,2H),2.62(t,2H),2.42–2.28(m,2H),2.17–1.86(m,5H) ,1.31–1.25(m,3H).

LCMS m/z = 510.6 [M+l].

Example 3

(S)-5-((1-(3-(4-(5-(1-fluorocyclopropyl)pyrimidin-2-yl)piperazin-1-yl)-3-oxopropoxy)propane -2-yl)amino)-4-(trifluoromethyl)pyridazin-3(2H)-one (compound 3)

(S)-5-((1-(3-(4-(5-(1-fluorocyclopropyl)pyrimidin-2-yl)piperazin-1-yl)-3-oxopropoxy)propan-2-yl)amino)- 4-(trifluoromethyl)pyridazin-3(2H)-one

Referring to the synthetic method of compound 1, compound 3 was prepared as a white solid (70 mg, yield 42%).

¹ H NMR (600MHz,DMSO-d ₆ )δ12.46(s,1H),8.45(d,2H),7.91(s,1H),6.30–6.26(m,1H),4.17–4.13(m,1H ),3.76(t,2H),3.72–3.65(m,4H),3.53–3.50(m,4H),3.48(d,2H),2.59(t,2H),1.40–1.32(m,2H), 1.15(d,3H),1.09–1.05(m,2H).

LCMS m/z = 514.4 [M+l].

Example 4

(S)-5-((1-(3-(4-(5-(1-hydroxycyclobutyl)pyrimidin-2-yl)piperazin-1-yl)-3-oxopropoxy)propane -2-yl)amino)-4-(trifluoromethyl)pyridazin-3(2H)-one (compound 4)

(S)-5-((1-(3-(4-(5-(1-hydroxycyclobutyl)pyrimidin-2-yl)piperazin-1-yl)-3-oxopropoxy)-propan-2-yl)amino) -4-(trifluoromethyl)pyridazin-3(2H)-one

first step:

1-(2-(piperazin-1-yl)pyrimidin-5-yl)cyclobutanol (4A)

1-(2-(piperazin-1-yl)pyrimidin-5-yl)cyclobutan-1-ol

Referring to the synthesis method of intermediate 2c, the crude product of compound 4A was obtained as a white solid. Compound 4A was directly used in the next reaction without further purification.

LCMS m/z = 235.2 [M+1].

Step two:

(S)-5-((1-(3-(4-(5-(1-hydroxycyclobutyl)pyrimidin-2-yl)piperazin-1-yl)-3-oxopropoxy)propane -2-yl)amino)-4-(trifluoromethyl)pyridazin-3(2H)-one (compound 4)

(S)-5-((1-(3-(4-(5-(1-hydroxycyclobutyl)pyrimidin-2-yl)piperazin-1-yl)-3-oxopropoxy)-propan-2-yl)amino) -4-(trifluoromethyl)pyridazin-3(2H)-one

Referring to the synthetic method of compound 1, compound 4 was prepared as a white solid (70 mg, yield 26%).

¹ H NMR (400MHz,DMSO-d ₆ )δ12.46(s,1H),8.47(s,2H),7.91(s,1H),6.31–6.27(m,1H),5.59(s,1H), 4.23–4.03(m,1H),3.74–3.64(m,6H),3.52–3.47(m,6H),2.58(t,2H),2.40–2.34(m,2H),2.27–2.20(m,2H ), 1.86–1.77(m,1H), 1.61–1.49(m,1H), 1.14(d,3H).

LCMS m/z = 526.3 [M+1].

Example 5

5-(((S)-1-(3-((2R,6S)-4-(5-cyclopropylpyrimidin-2-yl)-2,6-dimethylpiperazin-1-yl)- 3-oxopropoxy)propan-2yl)amino)-4-(trifluoromethyl)pyridazin-3(2H)-one (Compound 5)

5-(((S)-1-(3-((2R,6S)-4-(5-cyclopropylpyrimidin-2-yl)-2,6-dimethylpiperazin-1-yl)-3-oxopropoxy)propan-2 -yl)amino)-4-(trifluoromethyl)pyridazin-3(2H)-one

Referring to the synthetic method of compound 1, compound 5 was prepared as a white solid (280 mg, yield 53%).

¹ H NMR (400MHz,DMSO-d ₆ )δ12.45(s,1H),8.18(s,2H),7.91(s,1H),6.28(dd,1H),4.50(d,3H),4.23– 4.06(m,2H),3.69(s,2H),3.48(d,2H),3.03–2.85(m,2H),2.72–2.57(m,1H),2.42(s,2H),1.80–1.72( m,1H), 1.18–1.00(m,8H), 0.96–0.76(m,2H), 0.71–0.53(m,2H).

LCMS m/z = 524.3 [M+l].

Example 6

5-(((S)-1-(3-((2R,6S)-2,6-dimethyl-4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1- Carbonyl)cyclobutoxy)propan-2-yl)amino)-2-(4-methoxybenzyl)-4-(trifluoromethyl)pyridazin-3(2H)-one (compound 6)

N-(1-(5-cyclopropylpyrimidin-2-yl)-3,3-difluoropiperidin-4-yl)-3-((S)-3-((6-oxo-5-(trifluoromethyl)-1,6 -dihydropyridazin-4-yl)amino)butoxy)propanamide

Referring to the synthetic method of compound 1, compound 6 was prepared as a white solid (60 mg, yield 82%).

¹ H NMR (400MHz,DMSO-d ₆ )δ12.47(s,1H),8.20(s,2H),8.13(d,1H),7.90(s,1H),6.26(d,1H),4.84– 4.74(m,1H),4.47(dd,3H),4.12(s,2H),3.68–3.56(m,2H),3.48–3.42(m,2H),3.16(t,1H),2.40–2.37( m,2H),1.80–1.73(m,2H),1.60–1.49(m,1H),1.15(s,1H),1.13(s,1H),0.93–0.83(m,2H),0.67–0.61( m,2H).

LCMS m/z = 560.54 [M+l].

Example 7

(S)-5-((1-(3-(4-(5-(1-hydroxycyclopropyl)pyrimidin-2-yl)piperazin-1-yl)-3-oxopropoxy)propane -2-yl)amino)-4-(trifluoromethyl)pyridazin-3(2H)-one (compound 7)

(S)-5-((1-(3-(4-(5-(1-hydroxycyclopropyl)pyrimidin-2-yl)piperazin-1-yl)-3-oxopropoxy)propan-2-yl)amino)- 4-(trifluoromethyl)pyridazin-3(2H)-one

first step:

1-(2-(piperazin-1-yl)pyrimidin-5-yl)cyclopropan-1-ol (7A)

1-(2-(piperazin-1-yl)pyrimidin-5-yl)cyclopropan-1-ol

Referring to the synthesis method of intermediate 2c, the crude product of compound 7A was prepared, and compound 7A could be directly used in the next reaction without further purification.

LCMS m/z = 221.3 [M+l].

Step two:

(S)-5-((1-(3-(4-(5-(1-hydroxycyclopropyl)pyrimidin-2-yl)piperazin-1-yl)-3-oxopropoxy)propane -2-yl)amino)-4-(trifluoromethyl)pyridazin-3(2H)-one (compound 7)

(S)-5-((1-(3-(4-(5-(1-hydroxycyclopropyl)pyrimidin-2-yl)piperazin-1-yl)-3-oxopropoxy)propan-2-yl)amino)- 4-(trifluoromethyl)pyridazin-3(2H)-one

Referring to the synthetic method of compound 1, compound 7 was prepared as a white solid (16 mg, yield 6%).

¹ H NMR (400MHz,DMSO-d ₆ )δ12.46(s,1H),8.29(s,2H),7.91(s,1H),6.31–6.26(m1H),5.96(s,1H),4.20– 4.11(m,1H),3.85(d,1H),3.71–3.64(m,5H),3.54(d,1H),3.51–3.46(m,5H),2.58(t,2H),1.14(d, 3H), 1.02–0.96(m,2H), 0.90–0.85(m,2H).

LCMS m/z = 512.3 [M+l].

Example 8

(S)-5-((1-(3-(7-(5-cyclopropylpyrimidin-2-yl)-4,7-diazaspiro[2.5]sindan-4-yl)-3- Oxopropoxy)propan-2-yl)amino)-4-(trifluoromethyl)pyridazin-3(2H)-one (Compound 8)

(S)-5-((1-(3-(7-(5-cyclopropylpyrimidin-2-yl)-4,7-diazaspiro[2.5]octan-4-yl)-3-oxopropoxy)propan-2-yl )amino)-4-(trifluoromethyl)pyridazin-3(2H)-one

Referring to the synthetic method of compound 1, compound 8 was prepared as a white solid (40 mg, yield 25%).

¹ H NMR (400MHz,DMSO-d ₆ )δ12.44(s,1H),8.14(s,2H),7.89(s,1H),6.26–6.23(m,1H),4.13(t,1H), 3.81–3.40(m,9H),2.71(t,2H),2.52(s,1H),1.78–1.71(m,1H),1.11(d,3H),1.06–0.70(m,6H),0.65– 0.57(m,2H).

LCMS m/z = 522.3 [M+l].

Example 9

N-((1R, 5S)-3-(5-cyclopropylpyrimidin-2-yl)-3-azabicyclo[3.1.0]hexyl 6-yl)-3-((S)-2-( (6-oxo-5-(trifluoromethyl)-1,6-dihydropyridazin-4-yl)amino)propoxy)propionamide (Compound 9)

N-((1R,5S)-3-(5-cyclopropylpyrimidin-2-yl)-3-azabicyclo[3.1.0]hexan-6-yl)-3-((S)-2-((6-oxo -5-(trifluoromethyl)-1,6-dihydropyridazin-4-yl)amino)propoxy)propanamide

Referring to the synthesis method of compound 1, compound 9 was prepared as a white solid (76 mg, yield 50%).

¹ H NMR (600MHz,DMSO-d ₆ )δ12.47(s,1H),8.13(s,2H),8.02(d,1H),7.90(s,1H),6.27(dd,1H),4.13( t,1H),3.75(dd,2H),3.68–3.56(m,2H),3.45–3.41(m,4H),2.52(d,1H),2.33–2.31(m,1H),2.25(t, 2H), 1.80–1.66(m,3H), 1.14(d,3H), 0.92–0.80(m,2H), 0.65–0.54(m,2H).

LCMS m/z = 508.3 [M+l].

Example 10

(S)-5-((1-(2-(4-(5-cyclopropylpyrimidin-2-yl)piperazin-1-yl)-2-oxyethoxy)propan-2-yl)amino )-4-(trifluoromethyl)pyridazin 3(2H)-one (compound 10)

(S)-5-((1-(2-(4-(5-cyclopropylpyrimidin-2-yl)piperazin-1-yl)-2-oxoethoxy)propan-2-yl)amino)-4-(trifluoromethyl) pyridazin-3(2H)-one

first step:

(S)-5-((1-(2-(4-(5-cyclopropylpyrimidin-2-yl)piperazin-1-yl)-2-oxoethoxy)propan-2-yl) Amino)-2-(4-methoxybenzyl)-4-(trifluoromethyl)pyridazin-3(2H)-one (10A)

(S)-5-((1-(2-(4-(5-cyclopropylpyrimidin-2-yl)piperazin-1-yl)-2-oxoethoxy)propan-2-yl)amino)-2-(4- methoxybenzyl)-4-(trifluoromethyl)pyridazin-3(2H)-one

Referring to the synthetic method of compound 1, compound 10A was prepared as a white solid product (200 mg, yield 63%).

LC-MS m/z (ESI) = 602.6 [M+1].

Step two:

(S)-5-((1-(2-(4-(5-cyclopropylpyrimidin-2-yl)piperazin-1-yl)-2-oxyethoxy)propan-2-yl)amino )-4-(trifluoromethyl)pyridazin 3(2H)-one (compound 10)

(S)-5-((1-(2-(4-(5-cyclopropylpyrimidin-2-yl)piperazin-1-yl)-2-oxoethoxy)propan-2-yl)amino)-4-(trifluoromethyl) pyridazin-3(2H)-one

To a 25 mL reaction flask weighing compound 10A (200 mg, 0.33 mmol, 1.0 equiv) was added trifluoroacetic acid (1 mL) followed by trifluoromethanesulfonic acid (0.2 mL, 2.64 mmol, 8.0 equiv). After the addition was complete, the reaction was stirred at 25 °C for 2 h. Subsequently, 15 mL of water was added to the reaction solution to quench it. Extract with ethyl acetate (3×20 mL), and adjust the pH value of the organic layer to 8-9 with potassium carbonate aqueous solution. The organic layers were combined and concentrated in vacuo, and the residue was purified by reverse phase (water:acetonitrile=3:2) to obtain compound 10 as a white solid (105 mg, yield 62%).

¹ H NMR (600MHz,DMSO)δ12.47(s,1H),8.19(s,2H),7.94(s,1H),6.66(dd,1H),4.41–4.09(m,3H),3.76–3.61 (m,4H),3.54(d,2H),3.50(dd,2H),3.40(t,2H),1.82–1.71(m,1H),1.17(d,3H),0.93–0.81(m,2H ), 0.68–0.59(m,2H).

LC-MS m/z (ESI) = 482.2 [M+1].

Example 11

5-((S)-1-(3-((S)-4-(5-cyclopropylpyrimidin-2-yl)-2-(hydroxymethyl)piperazin-1-yl)-3-oxo Propoxy)propan-2-yl)amino)-4-(trifluoromethyl)pyridazin-3(2H)-one (compound 11)

5-(((S)-1-(3-((S)-4-(5-cyclopropylpyrimidin-2-yl)-2-(hydroxymethyl)piperazin-1-yl)-3-oxopropoxy)propan-2- yl)amino)-4-(trifluoromethyl)pyridazin-3(2H)-one

first step:

(S)-3-(Hydroxymethyl)-4-(3-((S)-2-((6-oxo-5-(trifluoromethyl)-1,6-dihydropyridazine-4 -yl)amino)propoxy)propionyl)piperazine-1-carboxylic acid tert-butyl ester (11B)

tert-butyl(S)-3-(hydroxymethyl)-4-(3-((S)-2-((6-oxo-5-(trifluoromethyl)-1,6-dihydropyridazin-4-yl)amino)propoxy )propanoyl)piperazine-1-carboxylate

Referring to the synthetic method of compound 1, compound 11B was prepared as a white solid (320 mg, 97%).

LC-MS m/z (ESI) = 508.3 [M+1].

Step two:

5-((S)-1-(3-((S)-2-(Hydroxymethyl)piperazin-1-yl)-3-oxopropoxy)propan-2-yl)amino)-4- (Trifluoromethyl)pyridazin-3(2H)-one (11C)

5-(((S)-1-(3-((S)-2-(hydroxymethyl)piperazin-1-yl)-3-oxopropoxy)propan-2-yl)amino)-4-(trifluoromethyl)pyridazin- 3(2H)-one

Referring to the synthesis method of intermediate 2c, the crude compound 11C was prepared. 11C can be directly used in the next reaction without further purification.

LC-MS m/z (ESI) = 408.3 [M+1].

third step:

5-((S)-1-(3-((S)-4-(5-cyclopropylpyrimidin-2-yl)-2-(hydroxymethyl)piperazin-1-yl)-3-oxo Propoxy)propan-2-yl)amino)-4-(trifluoromethyl)pyridazin-3(2H)-one (compound 11)

5-(((S)-1-(3-((S)-4-(5-cyclopropylpyrimidin-2-yl)-2-(hydroxymethyl)piperazin-1-yl)-3-oxopropoxy)propan-2- yl)amino)-4-(trifluoromethyl)pyridazin-3(2H)-one

Compound 11C (200 mg, 0.416 mmol, 1.0 equiv) was dissolved in N'N-dimethylacetamide (2.0 mL), and intermediate 7a (70.7 mg, 0.46 mmol, 1.1 equiv), N, N-di Isopropylethylamine (0.36 mL, 2.08 mmol, 5.0 equiv). After the addition was complete, the reaction was stirred at 120° C. for 5 h. After the reaction was complete, the reaction mixture was concentrated in vacuo. The residue was purified by C18 reverse phase chromatography (water:acetonitrile=1:1.5) to obtain compound 11 as a white solid (30 mg, 14%).

¹ H NMR(600MHz,DMSO-d6)δ12.47(d,1H),8.18(d,2H),7.92(s,1H),6.29–6.27(m,1H),4.63–4.42(m,2H) ,4.32–4.25(m,1H),4.17–4.12(m,1H),4.03(d,J＝9.6Hz,1H),3.67(t,2H),3.49(d,1H),3.44–3.34(m ,4H),3.30–3.20(m,1H),3.01–2.99(m,1H),2.80–2.69(m,2H),2.65–2.57(m,1H),1.78–1.74(m,1H),1.15 (d,3H), 0.95–0.79(m,2H), 0.69–0.57(m,2H).

LC-MS m/z (ESI) = 526.23 [M+1].

Example 12

(S)-1-(2-(4-(3-(2-(((6-oxo-5-(trifluoromethyl)-1,6-dihydropyridazin-4-yl)amino) Propoxy)propionyl)piperazine]-1-yl)pyrimidin-5-yl)cyclopropane-1-carbonitrile (compound 12)

(S)-1-(2-(4-(3-(2-((6-oxo-5-(trifluoromethyl)-1,6-dihydropyridazin-4-yl)amino)propoxy)propanoyl)piperazin-1- yl)pyrimidin-5-yl)cyclopropane-1-carbonitrile

Referring to the synthetic method of compound 1, compound 12 was prepared as a white solid (98 mg, yield 34%).

¹ H NMR (400MHz,DMSO-d ₆ )δ12.46(s,1H),8.30(s,2H),7.90(s,1H),6.34–6.28(m1H),5.96(s,1H),4.19– 4.10(m,1H),3.87(d,1H),3.73–3.62(m,5H),3.54–3.52(m,2H),3.50–3.44(m,4H),2.44(t,2H),1.20( d,3H), 1.00–0.95(m,2H), 0.90–0.87(m,2H).

LC-MS m/z = 521.50 [M+l].

Example 13

5-(((S)-1-(3-(4-(5-((R)-2,2-difluorocyclopropyl)]pyrimidin-2-yl)piperazin-1-yl)-3 -Oxopropoxy)propan-2-yl)amino)-4-(trifluoromethyl)pyridazin-3(2H)-one (compound 13-1)

5-(((S)-1-(3-(4-(5-((R)-2,2-difluorocyclopropyl)pyrimidin-2-yl)piperazin-1-yl)-3-oxopropoxy)propan-2 -yl)amino)-4-(trifluoromethyl)pyridazin-3(2H)-one

5-(((S)-1-(3-(4-(5-((S)-2,2-difluorocyclopropyl))pyrimidin-2-yl)piperidin-1-yl)-3 -propoxy)propan-2-yl)amino)-4-(trifluoromethyl)pyridazin-3(2H)-one (compound 13-2)

5-(((S)-1-(3-(4-(5-((S)-2,2-difluorocyclopropyl)pyrimidin-2-yl)piperazin-1-yl)-3-oxopropoxy)propan-2 -yl)amino)-4-(trifluoromethyl)pyridazin-3(2H)-one

first step:

5-(((S)-1-(3-(4-(5-(2,2-difluorocyclopropyl)]pyrimidin-2-yl)piperazin-1-yl)-3-oxopropane Oxy)propan-2-yl)amino)-4-(trifluoromethyl)pyridazin-3(2H)-one (compound 13)

5-(((S)-1-(3-(4-(5-(2,2-difluorocyclopropyl)pyrimidin-2-yl)piperazin-1-yl)-3-oxopropoxy)propan-2-yl)amino )-4-(trifluoromethyl)pyridazin-3(2H)-one

Referring to the synthetic method of compound 1, compound 13 was prepared as a white solid (98 mg, yield 34%).

LCMS m/z = 521.50 [M+l].

Step two:

5-(((S)-1-(3-(4-(5-((R)-2,2-difluorocyclopropyl)]pyrimidin-2-yl)piperazin-1-yl)-3 -Oxopropoxy)propan-2-yl)amino)-4-(trifluoromethyl)pyridazin-3(2H)-one (compound 13-1)

5-(((S)-1-(3-(4-(5-((R)-2,2-difluorocyclopropyl)pyrimidin-2-yl)piperazin-1-yl)-3-oxopropoxy)propan-2 -yl)amino)-4-(trifluoromethyl)pyridazin-3(2H)-one

5-(((S)-1-(3-(4-(5-((S)-2,2-difluorocyclopropyl))pyrimidin-2-yl)piperidin-1-yl)-3 -propoxy)propan-2-yl)amino)-4-(trifluoromethyl)pyridazin-3(2H)-one (compound 13-2)

5-(((S)-1-(3-(4-(5-((S)-2,2-difluorocyclopropyl)pyrimidin-2-yl)piperazin-1-yl)-3-oxopropoxy)propan-2 -yl)amino)-4-(trifluoromethyl)pyridazin-3(2H)-one

Racemate 13 was resolved by SFC to obtain compound 13-1 (27mg, ee%: 100%, chiral HPLC (OX-3); mobile phase: n-hexane:ethanol=70:30; column temperature: 35; column pressure : 80bar; flow rate: 2mL/min; detector signal channel: 214nm@4.8nm; diode array detector start wavelength: 200nm; diode array detector stop wavelength: 400nm; RT = 11.738min) and compound 13-2 (24mg , ee%: 97.18%, chiral HPLC (OX-3); Mobile phase: n-hexane: ethanol=70:30; Column temperature: 35; Column pressure: 80bar; Flow rate: 2mL/min; Detector signal channel: 214nm @4.8nm; diode array detector start wavelength: 200nm; diode array detector stop wavelength: 400nm; RT = 18.869min).

Compound 13-1: ¹ H NMR (400MHz, DMSO-d ₆ ) δ8.80(s,2H),7.87(s,1H),6.23(dd,1H),6.05(s,1H),5.00(s, 2H),4.45(dd,1H),4.36–4.26(m,2H),4.15–4.09(m,2H),3.33–3.18(m,2H),1.33(d,3H),1.12(d,3H) .

Compound 13-2: ¹ H NMR (400MHz, DMSO-d6) δ8.79(s, 2H), 7.86(s, 1H), 6.25(dd, 1H), 6.10(s, 1H), 5.08(s, 2H ), 4.43(dd,1H), 4.31–4.24(m,2H), 4.14–4.06(m,2H), 3.39–3.24(m,2H), 1.31(d,3H), 1.13(d,3H).

Example 14

5-(((S)-1-(3-oxo-3-(4-(5-((R)-1,2,2-trifluorocyclopropyl)pyrimidin-2-yl)piperazine- 1-yl)propoxy)propan-2-yl)amino)-4-(trifluoromethyl)pyridazine-3(2H)-one (compound 14-1)

5-(((S)-1-(3-oxo-3-(4-(5-((S)-1,2,2-trifluorocyclopropyl)pyrimidin-2-yl)piperazin-1-yl)propoxy) propan-2-yl)amino)-4-(trifluoromethyl)pyridazin-3(2H)-one

5-(((S)-1-(3-oxo-3-(4-(5-((S)-1,2,2-trifluorocyclopropyl)pyrimidin-2-yl)piperazine- 1-yl)propoxy)propan-2-yl)amino)-4-(trifluoromethyl)pyridazine-3(2H)-one (compound 14-2)

5-(((S)-1-(3-oxo-3-(4-(5-((R)-1,2,2-trifluorocyclopropyl)pyrimidin-2-yl)piperazin-1-yl)propoxy) propan-2-yl)amino)-4-(trifluoromethyl)pyridazin-3(2H)-one

first step:

5-(((S)-1-(3-oxo-3-(4-(5-(1,2,2-trifluorocyclopropyl)pyrimidin-2-yl)piperazin-1-yl) Propoxy)prop-2-yl)amino)-4-(trifluoromethyl)pyridazine-3(2H)-one (Compound 14)

5-(((2S)-1-(3-oxo-3-(4-(5-(1,2,2-trifluorocyclopropyl)pyrimidin-2-yl)piperazin-1-yl)propoxy)propan-2- yl)amino)-4-(trifluoromethyl)pyridazin-3(2H)-one

Referring to the synthetic method of compound 1, compound 14 was prepared as a white solid (51 mg, yield 46%).

LCMS m/z = 550.40 [M+l].

Step two:

5-(((S)-1-(3-oxo-3-(4-(5-((R)-1,2,2-trifluorocyclopropyl)pyrimidin-2-yl)piperazine- 1-yl)propoxy)propan-2-yl)amino)-4-(trifluoromethyl)pyridazine-3(2H)-one (compound 14-1)

5-(((S)-1-(3-oxo-3-(4-(5-((S)-1,2,2-trifluorocyclopropyl)pyrimidin-2-yl)piperazin-1-yl)propoxy) propan-2-yl)amino)-4-(trifluoromethyl)pyridazin-3(2H)-one

5-(((S)-1-(3-oxo-3-(4-(5-((S)-1,2,2-trifluorocyclopropyl)pyrimidin-2-yl)piperazine- 1-yl)propoxy)propan-2-yl)amino)-4-(trifluoromethyl)pyridazine-3(2H)-one (compound 14-2)

5-(((S)-1-(3-oxo-3-(4-(5-((R)-1,2,2-trifluorocyclopropyl)pyrimidin-2-yl)piperazin-1-yl)propoxy) propan-2-yl)amino)-4-(trifluoromethyl)pyridazin-3(2H)-one

Racemate 14 was resolved by SFC to obtain compound 14-1 (30mg, ee%: 100%, chiral HPLC (OX-3); mobile phase: n-hexane:ethanol=70:30; column temperature: 35; column pressure : 80bar; flow rate: 2mL/min; detector signal channel: 214nm@4.8nm; diode array detector start wavelength: 200nm; diode array detector stop wavelength: 400nm; RT = 15.850min) and compound 14-2 (29mg , ee%: 100%, chiral HPLC (OX-3); Mobile phase: n-hexane: ethanol=70:30; Column temperature: 35; Column pressure: 80bar; Flow velocity: 2mL/min; Detector signal channel: 214nm @4.8nm; diode array detector start wavelength: 200nm; diode array detector stop wavelength: 400nm; RT=16.936min).

Compound 14-1: ¹ H NMR (600MHz, DMSO-d ₆ ) δ12.46(s, 1H), 8.59(d, 2H), 7.92(s, 1H), 6.28(dd, 1H), 4.17-4.13( m,1H),3.80(t,2H),3.76(dd,2H),3.71–3.64(m,2H),3.53(t,4H),3.49(d,2H),2.59(t,3H),2.47 –2.38(m,1H),1.15(d,3H).

Compound 14-2: ¹ H NMR (600MHz, DMSO-d ₆ ) δ12.46(s,1H),8.59(d,2H),7.91(s,1H),6.28(dd,1H),4.15(t, 1H),3.80(t,2H),3.76(t,2H),3.72–3.63(m,2H),3.57–3.46(m,6H),2.67–2.57(m,3H),2.47–2.37(m, 1H), 1.15(d, 3H).

Example 15

5-((2S)-1-(3-(3-(5-cyclopropylpyrimidin-2-yl)-3,6-diazacyclo[3.1.1]heptane-6-yl)-3 -Oxypropoxy)propan-2-yl)amino)-4-(trifluoromethyl)pyridazin-3(2H)-one (compound 15)

5-(((2S)-1-(3-(3-(5-cyclopropylpyrimidin-2-yl)-3,6-diazabicyclo[3.1.1]heptan-6-yl)-3-oxopr opoxy)propan- 2-yl)amino)-4-(trifluoromethyl)pyridazin-3(2H)-one

Referring to the synthetic method of compound 1, compound 15 was prepared as a white solid (270 mg, yield 44%).

¹ H NMR (400MHz,DMSO-d ₆ )δ12.48(s,1H),8.18–8.14(m,2H),7.81(d,1H),6.20(s,1H),4.61(s,1H), 4.38–4.36(m,1H),4.00–3.96(m,1H),3.90–3.79(m,2H),3.70–3.65(m,1H),3.61–3.56(m,3H),3.41–3.73(m ,1H),2.59(d,1H),2.42–2.31(m,1H),2.29–2.20(m,1H),1.77–1.79(m,1H),1.53(d,1H),1.03–1.01(m ,3H), 0.87–0.82(m,2H),0.61–0.55(m,2H).

LCMS m/z = 508.10 [M+l].

Example 16

5-((2S)-1-(3-(8-(5-cyclopropylpyrimidin-2-yl)-3,8-diazacyclo[3.2.1]octane-3-yl)-3 -Oxypropoxy)propan-2-yl)amino)-4-(trifluoromethyl)pyridazin-3(2H)-one (compound 16)

5-(((2S)-1-(3-(8-(5-cyclopropylpyrimidin-2-yl)-3,8-diazabicyclo[3.2.1]octan-3-yl)-3-oxopropoxy)propan-2 -yl)amino)-4-(trifluoromethyl)pyridazin-3(2H)-one

Referring to the synthetic method of compound 1, compound 16 was prepared as a white solid (128 mg, yield 20%).

¹ H NMR (400MHz,DMSO-d ₆ )δ12.44(s,1H),8.17(d,2H),7.91(s,1H),6.28(s,1H),4.99–4.55(m,2H), 4.34–4.06(m,3H),3.68(t,2H),3.48(t,2H),3.20–3.08(m,1H),2.95(dd,1H),2.78–2.64(m,1H),1.81– 1.70(m,1H),1.14(d,3H),1.10(d,1H),1.02(dd,3H),0.96(d,1H),0.89–0.82(m,2H),0.65–0.59(m, 2H).

LCMS m/z = 522.10 [M+l].

Example 17

5-((2S)-1-(3-(3-(5-cyclopropylpyrimidin-2-yl)-3,8-diazacyclo[3.2.1]octane-8-yl)-3 -Oxypropoxy)propan-2-yl)amino)-4-(trifluoromethyl)pyridazin-3(2H)-one (compound 17)

5-(((2S)-1-(3-(3-(5-cyclopropylpyrimidin-2-yl)-3,8-diazabicyclo[3.2.1]octan-8-yl)-3-oxopropoxy)propan-2 -yl)amino)-4-(trifluoromethyl)pyridazin-3(2H)-one

Weigh 2-chloro-5-cyclopropylpyrimidine 7a (92mg, 0.6mmol, 1.0equiv) and dissolve it in N,N-dimethylformamide (10mL), add intermediate 15 (240mg, 0.6mmol, 1.0equiv ) and N,N-diisopropylethylamine (0.5mL, 3mmol, 5.0equiv), and the reaction was heated and stirred in an oil bath at 120°C for 1h. The reaction solution was cooled to room temperature and distilled under reduced pressure. The residue was purified by C18 reverse phase chromatography (water:acetonitrile=1:5) to obtain compound 17 as a white solid (78 mg, yield 25%).

¹ H NMR (400MHz, DMSO-d6) δ12.46(s,1H),8.15(d,2H),7.89(d,1H),6.26(dd,1H),4.59(d,1H),4.46–4.08 (m,6H),3.71–3.63(m,2H),3.48(d,2H),2.98(dd,1H),2.94–2.82(m,1H),2.62–2.52(m,1H),1.90–1.81 (m,1H), 1.67(d,1H), 1.65–1.45(m,2H), 1.12(dd,3H), 0.90–0.79(m,2H), 0.68–0.54(m,2H).

LC-MS m/z (ESI) = 522.20 [M+1].

Example 18

5-((S)-1-(3-((2R,5S)-4-(5-cyclopropylpyrimidin-2-yl)-2,5-dimethylpiperazin-1-yl)-3 -Oxypropoxy)propan-2-yl)amino)-4-(trifluoromethyl)pyridazin-3(2H)-one (compound 18)

5-(((S)-1-(3-((2R,5S)-4-(5-cyclopropylpyrimidin-2-yl)-2,5-dimethylpiperazin-1-yl)-3-oxopropoxy)propan-2 -yl)amino)-4-(trifluoromethyl)pyridazin-3(2H)-one

Referring to the synthesis method of compound 1, compound 18 was prepared as a white solid (40 mg, yield 50%).

¹ H NMR (400MHz,DMSO-d ₆ )δ12.44(s,1H),8.17(d,2H),7.91(s,1H),6.28(s,1H),4.89–4.59(m,2H), 4.26(d,1H),4.24–4.06(m,2H),3.68(t,2H),3.48(t,2H),3.44–3.40(m,1H),3.20–3.16(m,1H),3.12– 3.01(m,1H),2.98–2.93(m,1H),1.85–1.67(m,1H),1.14(d,3H),δ1.12–0.94(m,6H),0.90–0.82(m,2H ),0.67–0.56(m,2H).

LC-MS m/z (ESI) = 524.20 [M+1].

Example 19

5-((2S)-1-(3-(5-(5-cyclopropylpyrimidin-2-yl)-2,5-diazacyclo[2.2.1]heptane-2-yl)-3 -Oxypropoxy)propan-2-yl)amino)-4-(trifluoromethyl)pyridazin-3(2H)-one (compound 19)

5-(((2S)-1-(3-(5-(5-cyclopropylpyrimidin-2-yl)-2,5-diazabicyclo[2.2.1]heptan-2-yl)-3-oxopropoxy)propan-2 -yl)amino)-4-(trifluoromethyl)pyridazin-3(2H)-one

Referring to the synthetic method of compound 1, compound 19 was prepared as a white solid (80 mg, yield 31%).

¹ H NMR (400MHz,DMSO-d ₆ )δ12.47(s,1H),8.13(s,2H),7.85(d,1H),6.28–6.21(m,1H),4.86–4.69(m,2H ),4.06(d,1H),3.69–3.56(m,2H),3.53(d,1H),3.49–3.44(m,2H),3.39(d,2H),3.31–3.27(m,1H), 2.65–2.53(m,2H),2.36–2.25(m,1H),1.89(d,1H),1.77–1.70(m,1H),1.13–1.01(m,3H),0.86–0.83(m,2H ), 0.63–0.51(m,2H).

LC-MS m/z (ESI) = 508.20 [M+1].

Example 20

5-((S)-1-(3-((S)-4-(5-cyclopropylpyrimidin-2-yl)-2-methylpiperazin-1-yl)-3-oxopropoxy )propan-2-yl)amino)-4-(trifluoromethyl)pyridazin-3(2H)-one (compound 20-1)

5-(((S)-1-(3-((S)-4-(5-cyclopropylpyrimidin-2-yl)-2-methylpiperazin-1-yl)-3-oxopropoxy)propan-2-yl)amino )-4-(trifluoromethyl)pyridazin-3(2H)-one

5-((S)-1-(3-((R)-4-(5-cyclopropylpyrimidin-2-yl)-2-methylpiperazin-1-yl)-3-oxopropoxy )propan-2-yl)amino)-4-(trifluoromethyl)pyridazin-3(2H)-one (compound 20-2)

5-(((S)-1-(3-((R)-4-(5-cyclopropylpyrimidin-2-yl)-2-methylpiperazin-1-yl)-3-oxopropoxy)propan-2-yl)amino )-4-(trifluoromethyl)pyridazin-3(2H)-one

first step:

tert-butyl 3-methyl-4-(3-((S)-2-((6-oxo-5-(trifluoromethyl)-1,6-dihydropyridazin-4-yl)amino )propoxy)piperazine-1-carboxylate (20B)

tert-butyl 3-methyl-4-(3-((S)-2-((6-oxo-5-(trifluoromethyl)-1,6-dihydropyridazin-4-yl)amino)propoxy)propanoyl)piperazine-1 -carboxylate

Referring to the synthetic method of compound 1, compound 20B was prepared as a white solid (230 mg, yield 72%).

LC-MS m/z (ESI) = 492.51 [M+1].

Step two:

5-((2S)-1-(3-(2-Methylpiperazin-1-yl)-3-oxopropoxy)propan-2-yl)amino)-4-(trifluoromethyl)pyridium Azin-3(2H)-one (20C)

5-(((2S)-1-(3-(2-methylpiperazin-1-yl)-3-oxopropoxy)propan-2-yl)amino)-4-(trifluoromethyl)pyridazin-3(2H)-one

Referring to the synthesis method of intermediate 2c, the crude compound 20C was prepared as a white solid. Compound 20C was directly used in the next reaction without further purification.

LC-MS m/z (ESI) = 392.40 [M+1].

third step:

5-((2S)-1-(3-(4-(5-cyclopropylpyrimidin-2-yl)-2-methylpiperazin-1-yl)-3-oxopropoxy)propane-2 -yl)amino)-4-(trifluoromethyl)pyridazin-3(2H)-one (compound 20)

5-(((2S)-1-(3-(4-(5-cyclopropylpyrimidin-2-yl)-2-methylpiperazin-1-yl)-3-oxopropoxy)propan-2-yl)amino)-4- (trifluoromethyl)pyridazin-3(2H)-one

Referring to the synthetic method of compound 19, compound 20 was prepared as a white solid (140 mg, yield 40%).

LC-MS m/z (ESI) = 510.53 [M+1].

the fourth step:

5-((S)-1-(3-((S)-4-(5-cyclopropylpyrimidin-2-yl)-2-methylpiperazin-1-yl)-3-oxopropoxy )propan-2-yl)amino)-4-(trifluoromethyl)pyridazin-3(2H)-one (compound 20-1)

5-(((S)-1-(3-((S)-4-(5-cyclopropylpyrimidin-2-yl)-2-methylpiperazin-1-yl)-3-oxopropoxy)propan-2-yl)amino )-4-(trifluoromethyl)pyridazin-3(2H)-one

5-((S)-1-(3-((R)-4-(5-cyclopropylpyrimidin-2-yl)-2-methylpiperazin-1-yl)-3-oxopropoxy )propan-2-yl)amino)-4-(trifluoromethyl)pyridazin-3(2H)-one (compound 20-2)

5-(((S)-1-(3-((R)-4-(5-cyclopropylpyrimidin-2-yl)-2-methylpiperazin-1-yl)-3-oxopropoxy)propan-2-yl)amino )-4-(trifluoromethyl)pyridazin-3(2H)-one

Racemate compound 20 was resolved by SFC to obtain compound 20-1 (35 mg, ee%: 99.74%, chiral HPLC (OX-3); mobile phase: n-hexane:isopropanol=80:20; column temperature: 35 ; Column pressure: 80bar; Flow rate: 2mL/min; Detector signal channel: 214nm@4.8nm; Diode array detector start wavelength: 200nm; Diode array detector stop wavelength: 400nm; RT=8.157min) and compound 20- 2(29mg, ee%: 98.80%, chiral HPLC (OX-3); mobile phase: n-hexane:isopropanol=80:20; column temperature: 35; column pressure: 80bar; flow rate: 2mL/min; detection signal channel: 214nm@4.8nm; diode array detector start wavelength: 200nm; diode array detector stop wavelength: 400nm; RT=6.596min).

Compound 20-1: ¹ H NMR (400MHz, DMSO-d ₆ ) δ12.46(s, 1H), 8.17(s, 2H), 7.91(s, 1H), 6.29–6.26(m, 1H), 4.61( s,1H),4.48(d,1H),4.35(d,2H),4.23(d,1H),4.15(t,2H),3.76(d,1H),3.66(t,2H),3.47(d ,2H),3.05(t,1H),3.00–2.89(m,1H),2.83(q,1H),1.79–1.72(m,1H),1.14(d,3H),1.07(d,1H), 0.98(d,1H), 0.89–0.83(m,2H), 0.65–0.60(m,2H).

LC-MS m/z (ESI) = 510.60 [M+1].

Compound 20-2: ¹ H NMR (400MHz, DMSO-d ₆ ) δ12.46(s, 1H), 8.17(s, 2H), 7.91(s, 1H), 6.29–6.26(m, 1H), 4.62( s,1H),4.48(d,1H),4.37(t,,2H),4.23(d,,1H),4.14(t,2H),3.75(d,1H),3.66(d,2H),3.47 (d,2H),3.04(s,1H),2.94(s,1H),2.81(t,1H),1.79–1.72(m,1H),1.14(d,3H),1.07(d,1H), 0.98(d,1H), 0.89–0.83(m,2H), 0.66–0.60(m,2H).

Example 21

5-((S)-1-(3-((2S,5R)-4-(5-cyclopropylpyrimidin-2-yl)-2,5-dimethylpiperazin-1-yl)-3 -Oxypropoxy)propan-2-yl)amino)-4-(trifluoromethyl)pyridazin-3(2H)-one (compound 21)

5-(((S)-1-(3-((2S,5R)-4-(5-cyclopropylpyrimidin-2-yl)-2,5-dimethylpiperazin-1-yl)-3-oxopropoxy)propan-2 -yl)amino)-4-(trifluoromethyl)pyridazin-3(2H)-one

first step:

tert-butyl(2R,5S)-2,5-dimethyl-4-(3-((S)-2-((6-oxo-5-(trifluoromethyl)-1,6-di Hydropyridazin-4-yl)amino)propoxy)propionyl)piperazine-1-carboxylate (21B)

tert-butyl(2R,5S)-2,5-dimethyl-4-(3-((S)-2-((6-oxo-5-(trifluoromethyl)-1,6-dihydropyridazin-4-yl)amino )propoxy)propanoyl)piperazine-1-carboxylate

Referring to the synthetic method of compound 1, compound 21B was prepared as a white solid (120 mg, yield 61%).

LC-MS m/z (ESI) = 506.54 [M+1].

Step two:

5-((S)-1-(3-((2S,5R)-2,5-dimethylpiperazin-1-yl)-3-oxopropoxy)propan-2-yl)amino)- 4-(Trifluoromethyl)pyridazin-3(2H)-one (21C)

5-(((S)-1-(3-((2S,5R)-2,5-dimethylpiperazin-1-yl)-3-oxopropoxy)propan-2-yl)amino)-4-(trifluoromethyl)pyridazin -3(2H)-one

Referring to the synthesis method of intermediate 2c, the crude compound 21C was prepared as a white solid. Compound 21C can be directly used in the next reaction without further purification.

LC-MS m/z (ESI) = 406.42 [M+1].

third step:

5-((S)-1-(3-((2S,5R)-4-(5-cyclopropylpyrimidin-2-yl)-2,5-dimethylpiperazin-1-yl)-3 -Oxypropoxy)propan-2-yl)amino)-4-(trifluoromethyl)pyridazin-3(2H)-one (compound 21)

5-(((S)-1-(3-((2S,5R)-4-(5-cyclopropylpyrimidin-2-yl)-2,5-dimethylpiperazin-1-yl)-3-oxopropoxy)propan-2 -yl)amino)-4-(trifluoromethyl)pyridazin-3(2H)-one

Referring to the synthetic method of compound 19, compound 21 was prepared as a white solid (40 mg, yield 23%).

¹ H NMR(400MHz,DMSO-d6)δ12.45(s,1H),8.17(d,2H),7.91(s,1H),6.32–6.25(m,1H),4.85–4.66(m,2H) ,4.25(s,1H),4.24–4.11(m,2H),3.74–3.63(m,2H),3.48(t,2H),3.41(m,1H),3.22–3.15(m,1H),3.10 (m,1H),2.95(m,1H),1.74(m,1H),1.14(m,3H),1.11–0.94(m,6H),0.86(m,2H),0.66–0.59(m,2H ).

LC-MS m/z (ESI) = 524.56 [M+1].

Example 22

N-((3R,4R)-1-(5-cyclopropylpyrimidin-2-yl)-3-hydroxypiperidin-4-yl)-2-((S)-2-((6-oxy -5-(trifluoromethyl)-1,6-dihydropyridazin-4-yl)amino)propionamide (compound 22)

N-((3R,4R)-1-(5-cyclopropylpyrimidin-2-yl)-3-hydroxypiperidin-4-yl)-2-((S)-2-((6-oxo-5-(trifluoromethyl) -1,6-dihydropyridazin-4-yl)amino)propoxy)propanamide

first step:

tert-butyl(3R,4R)-3-hydroxy-4-(2-((S)-2-((1-(4-methoxybenzyl)-6-oxo-5-(trifluoromethyl base)-1,6-dihydropyridazin-4-yl)amino)propoxy)propionamide)piperidine-1-carboxylate (22B)

tert-butyl(3R,4R)-3-hydroxy-4-(2-((S)-2-((1-(4-methoxybenzyl)-6-oxo-5-(trifluorometh-yl)-1,6 -dihydropyridazin-4-yl)amino)propoxy)propanamido)piperidine-1-carboxylate

Referring to the synthetic method of compound 1, compound 22B was prepared as a white solid (220 mg, yield 60%).

LC-MS m/z (ESI) = 628.66 [M+1].

Step two:

N-((3R,4R)-3-hydroxypiperidin-4-yl)-2-((S)-2-((1-(4-methoxybenzyl)-6-oxyl-5- (Trifluoromethyl)-1,6-dihydropyridazin-4-yl)amino)propoxy)amide (22C)

N-((3R,4R)-3-hydroxypiperidin-4-yl)-2-((S)-2-((1-(4-methoxybenzyl)-6-oxo-5-(trifluoromethyl)-1,6 -dihydropyridazin-4-yl)amino)propoxy)propanamide

Referring to the synthetic method of compound 2c, the crude compound 22C was prepared as a white solid. Compound 22C can be directly used in the next reaction without further purification.

LC-MS m/z (ESI) = 528.55 [M+1].

third step:

N-((3R,4R)-1-(5-cyclopropylpyrimidin-2-yl)-3-hydroxypiperidin-4-yl)-2-((S)-2-((1-(4 -methoxybenzyl)-6-oxyl-5-(trifluoromethyl)-1,6-dihydropyridazin-4-yl)amino)propoxy)propylamine (22D)

N-((3R,4R)-1-(5-cyclopropylpyrimidin-2-yl)-3-hydroxypiperidin-4-yl)-2-((S)-2-((1-(4-methoxybenzyl)-6 -oxo-5-(trifluoromethyl)-1,6-dihydropyridazin-4-yl)amino)propoxy)propanamide

Referring to the synthetic method of compound 19, compound 22D was prepared as a white solid (110 mg, yield 45%).

LC-MS m/z (ESI) = 646.68 [M+1].

the fourth step:

N-((3R,4R)-1-(5-cyclopropylpyrimidin-2-yl)-3-hydroxypiperidin-4-yl)-2-((S)-2-((6-oxy -5-(trifluoromethyl)-1,6-dihydropyridazin-4-yl)amino)propionamide (compound 22)

N-((3R,4R)-1-(5-cyclopropylpyrimidin-2-yl)-3-hydroxypiperidin-4-yl)-2-((S)-2-((6-oxo-5-(trifluoromethyl) -1,6-dihydropyridazin-4-yl)amino)propoxy)propanamide

Referring to the synthesis method of intermediate 1i, compound 22 was prepared as a white solid (41 mg, yield 45%).

¹ H NMR (400MHz, DMSO-d6) δ12.45 (d, J = 24.8Hz, 1H), 8.16 (s, 2H), 7.95 (d, 1H), 7.55–7.52 (m, 1H), 6.49–6.29 (m,1H),4.68–4.43(m,2H),4.16(s,1H),3.85–3.78(m,1H),3.72–3.64(m,1H),3.46–3.29(m,5H),2.78 (d,2H), 1.79–1.73(m,2H), 1.20–1.14(m,6H), 0.90–0.81(m,2H), 0.65–0.57(m,2H).

LC-MS m/z (ESI) = 526.53 [M+1].

Example 23

(R)-5-(2-((3-(4-(5-cyclopropylpyrimidin-2-yl)piperazin-1-yl)-3-oxopropoxy)methyl)pyrrolidine-1 -yl)-4-(trifluoromethyl)pyridazin-3(2H)-one (compound 23-1)

(R)-5-(2-((3-(4-(5-cyclopropylpyrimidin-2-yl)piperazin-1-yl)-3-oxopropoxy)methyl)pyrrolidin-1-yl)-4-(trifluoromethyl) pyridazin-3(2H)-one

(S)-5-(2-((3-(4-(5-cyclopropylpyrimidin-2-yl)piperazin-1-yl)-3-oxopropoxy)methyl)pyrrolidine-1 -yl)-4-(trifluoromethyl)pyridazin-3(2H)-one (compound 23-2)

(S)-5-(2-((3-(4-(5-cyclopropylpyrimidin-2-yl)piperazin-1-yl)-3-oxopropoxy)methyl)pyrrolidin-1-yl)-4-(trifluoromethyl) pyridazin-3(2H)-one

first step:

2-(4-methoxybenzyl)-5-(2-((3-oxo-3-(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazin-1-yl )propoxy)methyl)pyrrolidin-1-yl)-4-(trifluoromethyl)pyridazin-3(2H)-one (23A)

2-(4-methoxybenzyl)-5-(2-((3-oxo-3-(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazin-1-yl)propoxy)methyl)pyrrolidin-1- yl)-4-(trifluoromethyl)pyridazin-3(2H)-one

Referring to the synthetic method of compound 1, compound 23A was prepared as a white solid (810 mg, yield 66%).

LC-MS m/z (ESI) = 670.25 [M+1].

Step two:

5-(2-((3-oxo-3-(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazin-1-yl)propoxy)methyl)pyrrolidine-1 -yl)-4-(trifluoromethyl)pyridazin-3(2H)-one (compound 23)

5-(2-((3-oxo-3-(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazin-1-yl)propoxy)methyl)pyrrolidin-1-yl)-4-(trifluoromethyl) pyridazin-3(2H)-one

Referring to the synthesis method of intermediate 1i, compound 23 was prepared as a white solid (485 mg, yield 73%).

LC-MS m/z (ESI) = 550.19 [M+1].

third step:

(R)-5-(2-((3-(4-(5-cyclopropylpyrimidin-2-yl)piperazin-1-yl)-3-oxopropoxy)methyl)pyrrolidine-1 -yl)-4-(trifluoromethyl)pyridazin-3(2H)-one (compound 23-1)

(R)-5-(2-((3-(4-(5-cyclopropylpyrimidin-2-yl)piperazin-1-yl)-3-oxopropoxy)methyl)pyrrolidin-1-yl)-4-(trifluoromethyl) pyridazin-3(2H)-one

(S)-5-(2-((3-(4-(5-cyclopropylpyrimidin-2-yl)piperazin-1-yl)-3-oxopropoxy)methyl)pyrrolidine-1 -yl)-4-(trifluoromethyl)pyridazin-3(2H)-one (compound 23-2)

(S)-5-(2-((3-(4-(5-cyclopropylpyrimidin-2-yl)piperazin-1-yl)-3-oxopropoxy)methyl)pyrrolidin-1-yl)-4-(trifluoromethyl) pyridazin-3(2H)-one

Racemate compound 23 was resolved by SFC to obtain compound 23-1 (54mg, ee%: 100%, chiral HPLC (OX-3); mobile phase: n-hexane:ethanol=70:30; column temperature: 35; Pressure: 80bar; Flow rate: 2mL/min; Detector signal channel: 214nm@4.8nm; Diode array detector start wavelength: 200nm; Diode array detector stop wavelength: 400nm; RT=11.738min) and compound 23-2( 56mg, ee%: 97.18%, chiral HPLC (OX-3); Mobile phase: n-hexane: ethanol=70:30; Column temperature: 35; Column pressure: 80bar; Flow rate: 2mL/min; Detector signal channel: 214nm@4.8nm; diode array detector start wavelength: 200nm; diode array detector stop wavelength: 400nm; RT=18.869min).

Compound 23-1: ¹ H NMR (400MHz, DMSO-d6) δ12.36(s,1H),8.19(s,2H),8.01(s,1H),4.56–4.47(m,1H),3.69–3.58 (m,6H),3.52–3.44(m,7H),3.22–3.16(m,1H),2.55–2.52(m,2H),2.06(q,J＝6.4Hz,1H),1.87(d,J =5.1Hz, 1H), 1.76(d, J=5.2Hz, 1H), 1.62(d, J=3.8Hz, 2H), 0.90–0.83(m, 2H), 0.66–0.60(m, 2H).

LC-MS m/z (ESI) = 550.19 [M+1].

Compound 23-2: ¹ H NMR (400MHz, DMSO-d6) δ12.36(s,1H),8.19(s,2H),8.01(s,1H),4.53–4.50(m,1H),3.70–3.57 (m,6H),3.52–3.44(m,7H),3.22–3.18(m,1H),2.55–2.51(m,2H),2.10–2.03(m,1H),1.87(d,J＝5.0Hz , 1H), 1.82–1.72(m, 1H), 1.62(t, J=5.2Hz, 2H), 0.90–0.83(m, 2H), 0.66–0.60(m, 2H).

LC-MS m/z (ESI) = 550.19 [M+1].

Example 24

5-((R)-2-((3-((2S,6R)-4-(5-cyclopropylpyrimidin-2-yl)-2,6-dimethylpiperazin-1-yl)- 3-oxopropoxy)methyl)pyrrolidin-1-yl)-4-(trifluoromethyl)pyridazin-3(2H)-one (compound 24-1)

5-((R)-2-((3-((2S,6R)-4-(5-cyclopropylpyrimidin-2-yl)-2,6-dimethylpiperazin-1-yl)-3-oxopropoxy)methyl)pyrrolidin -1-yl)-4-(trifluoromethyl)pyridazin-3(2H)-one

5-((S)-2-((3-((2S,6R)-4-(5-cyclopropylpyrimidin-2-yl)-2,6-dimethylpiperazin-1-yl)- 3-oxopropoxy)methyl)pyrrolidin-1-yl)-4-(trifluoromethyl)pyridazin-3(2H)-one (compound 24-2)

5-((S)-2-((3-((2S,6R)-4-(5-cyclopropylpyrimidin-2-yl)-2,6-dimethylpiperazin-1-yl)-3-oxopropoxy)methyl)pyrrolidin -1-yl)-4-(trifluoromethyl)pyridazin-3(2H)-one

first step:

5-(2-((3-((2R,6S)-4-(5-cyclopropylpyrimidin-2-yl)-2,6-dimethylpiperazin-1-yl)-3-oxopropane Oxy)methyl)pyrrolidin-1-yl)-2-(4-methoxybenzyl)-4-(trifluoromethyl)pyridazin-3(2H)-one (24A)

5-(2-((3-((2R,6S)-4-(5-cyclopropylpyrimidin-2-yl)-2,6-dimethylpiperazin-1-yl)-3-oxopropoxy)methyl)pyrrolidin-1-yl )-2-(4-methoxybenzyl)-4-(trifluoromethyl)pyridazin-3(2H)-one

Referring to the synthetic method of compound 1, compound 24A was prepared as a yellow oil (300 mg, yield 45%).

LCMS m/z = 670.33 [M+l].

Step two:

5-(2-((3-((2R,6S)-4-(5-cyclopropylpyrimidin-2-yl)-2,6-dimethylpiperazin-1-yl)-3-oxopropane Oxy)methyl)pyrrolidin-1-yl)-4-(trifluoromethyl)pyridazin-3(2H)-one (compound 24)

5-(2-((3-((2R,6S)-4-(5-cyclopropylpyrimidin-2-yl)-2,6-dimethylpiperazin-1-yl)-3-oxopropoxy)methyl)pyrrolidin-1-yl )-4-(trifluoromethyl)pyridazin-3(2H)-one

Referring to the synthesis method of intermediate 1i, compound 24 was prepared as a white solid (180 mg, yield 73%).

LC-MS m/z (ESI) = 550.27 [M+1].

third step:

5-((R)-2-((3-((2S,6R)-4-(5-cyclopropylpyrimidin-2-yl)-2,6-dimethylpiperazin-1-yl)- 3-oxopropoxy)methyl)pyrrolidin-1-yl)-4-(trifluoromethyl)pyridazin-3(2H)-one (compound 24-1)

5-((R)-2-((3-((2S,6R)-4-(5-cyclopropylpyrimidin-2-yl)-2,6-dimethylpiperazin-1-yl)-3-oxopropoxy)methyl)pyrrolidin -1-yl)-4-(trifluoromethyl)pyridazin-3(2H)-one

5-((S)-2-((3-((2S,6R)-4-(5-cyclopropylpyrimidin-2-yl)-2,6-dimethylpiperazin-1-yl)- 3-oxopropoxy)methyl)pyrrolidin-1-yl)-4-(trifluoromethyl)pyridazin-3(2H)-one (compound 24-2)

5-((S)-2-((3-((2S,6R)-4-(5-cyclopropylpyrimidin-2-yl)-2,6-dimethylpiperazin-1-yl)-3-oxopropoxy)methyl)pyrrolidin -1-yl)-4-(trifluoromethyl)pyridazin-3(2H)-one

Racemate compound 24 was resolved by SFC to obtain compound 24-1 (54mg, ee%: 100%, chiral HPLC (OX-3); mobile phase: n-hexane:ethanol=70:30; column temperature: 35; Pressure: 80bar; Flow rate: 2mL/min; Detector signal channel: 214nm@4.8nm; Diode array detector start wavelength: 200nm; Diode array detector stop wavelength: 400nm; RT=5.05min) and compound 24-2 ( 56mg, ee%: 97.18%, chiral HPLC (OX-3); Mobile phase: n-hexane: ethanol=70:30; Column temperature: 35; Column pressure: 80bar; Flow rate: 2mL/min; Detector signal channel: 214nm@4.8nm; diode array detector start wavelength: 200nm; diode array detector stop wavelength: 400nm; RT=7.49min).

Compound 24-1: ¹ H NMR (400MHz, DMSO-d ₆ ) δ12.34(s,1H),8.17(s,2H),8.01(s,1H),4.50(d,3H),4.19–4.03( m,1H),3.67(s,2H),3.52–3.49(m,2H),3.40–3.54(m,2H),3.21(s,1H),2.95(s,2H),2.45–2.37(m, 2H),2.07(s,1H),1.87(s,1H),1.79–1.72(m,1H),1.63(d,2H),1.08(d,6H),0.90–0.83(m,2H),0.66 –0.58(m,2H).

Compound 24-1: ¹ H NMR (400MHz, DMSO-d ₆ )δ12.34(s,1H),8.17(s,2H),8.01(s,1H),4.50(d,3H),4.09(s, 1H),3.66(s,2H),3.51(d,2H),3.43–3.34(m,2H),3.20(s,1H),2.96(s,2H),2.45–2.34(m,2H),2.07 (s,1H),1.87(s,1H),1.81–1.70(m,1H),1.62(s,2H),1.08(d,6H),0.90–0.82(m,2H),0.66–0.62(m ,2H).

Example 25

(S)-5-((1-(3-(4-(5-cyclopropylpyrimidin-2-yl)piperazin-1-yl)-3-oxopropoxy)propan-2-yl) Amino)-4-(trifluoromethyl)pyridazin 3(2H)-one (compound 25)

(S)-5-((1-(3-(4-(5-cyclopropylpyrimidin-2-yl)piperazin-1-yl)-3-oxopropoxy)propan-2-yl)amino)-4-(trifluoromethyl) pyridazin-3(2H)-one

Referring to the synthetic method of compound 1, compound 25 was prepared as a white solid (120 mg, yield 43%).

¹ H NMR (400MHz,DMSO-d ₆ )δ12.46(s,1H),8.36(s,1H),7.91(s,1H),7.72(s,1H),6.27(dd,1H),3.70- 3.65(m,2H),3.56–353(m,4H),3.48(d,2H),3.01-2.98(m,4H),2.58(t,2H),2.07–1.95(m,1H),1.15( d,3H), 0.98-0.90(m,2H), 0.84–0.73(m,2H).

LC-MS m/z (ESI) = 563.20 [M+1].

Example 26

(S)-5-cyclopropyl-2-(4-(3-(2-((6-oxo-5-(trifluoromethyl)-1,6-dihydropyridazin-4-yl) Amino)propoxy)propyl)piperazin-1-yl)nicotinonitrile (compound 26)

(S)-5-cyclopropyl-2-(4-(3-(2-((6-oxo-5-(trifluoromethyl)-1,6-dihydropyridazin-4-yl)amino)propoxy)propanoyl)piperazin-1 -yl)nicotinonitrole

Referring to the synthetic method of compound 1, compound 26 was prepared as a white solid (68 mg, yield 26%).

¹ H NMR (400MHz,DMSO-d ₆ )δ12.45(s,1H),8.29(s,1H),7.91(s,1H),7.79(s,1H),6.28(dd,1H),4.21– 4.09(m,2H),3.68-3.61(m,4H),3.58(d,2H),3.49-3.37(m,4H),2.59(t,2H),1.96-1.92(m,1H),1.15( d,3H), 0.98–0.88(m,2H), 0.77–0.66(m,2H).

LC-MS m/z (ESI) = 520.22 [M+1].

Example 27

(S)-5-((1-(3-(4-(5-cyclopropylpyrazin-2-yl)piperazin-1-yl)-3-oxopropoxy)propan-2-yl) Amino)-4-(trifluoromethyl)pyridazin-3(2H)-one (compound 27)

(S)-5-((1-(3-(4-(5-cyclopropylpyrazin-2-yl)piperazin-1-yl)-3-oxopropoxy)propan-2-yl)amino)-4-(trifluoromethyl) pyridazin-3(2H)-one

Referring to the synthetic method of compound 1, compound 27 was prepared as a white solid (76 mg, yield 32%).

¹ H NMR (400MHz,DMSO-d ₆ )δ12.43(s,1H),8.26(s,1H),7.65(s,1H),7.58(s,1H),6.24(dd,1H),4.33– 4.26(m,2H),3.74–3.63(m,4H),3.52(d,2H),3.55-3.46(m,4H),2.59(t,2H),1.99-1.90(m,1H),1.15( d,3H), 0.96–0.84(m,2H), 0.79–0.63(m,2H).

LC-MS m/z (ESI) = 496.20 [M+1].

Example 28

5-(((2S)-1-((4-(4-(5-cyclopropylpyrimidin-2-yl)piperazin-1-yl)-4-oxobutan-2-yl)oxy) Propan-2-yl)amino)-4-(trifluoromethyl)pyridazin-3(2H)-one (compound 28)

5-(((2S)-1-((4-(4-(5-cyclopropylpyrimidin-2-yl)piperazin-1-yl)-4-oxobutan-2-yl)oxy)propan-2-yl)amino )-4-(trifluoromethyl)pyridazin-3(2H)-one

first step:

5-(((2S)-1-((4-(4-(5-cyclopropylpyrimidin-2-yl)piperazin-1-yl)-4-oxobutan-2-yl)oxy) Propan-2-yl)amino)-2-(4-methoxybenzyl)-4-(trifluoromethyl)pyridazin-3(2H)-one (28A)

5-(((2S)-1-((4-(4-(5-cyclopropylpyrimidin-2-yl)piperazin-1-yl)-4-oxobutan-2-yl)oxy)propan-2-yl)amino )-2-(4-methoxybenzyl)-4-(trifluoromethyl)pyridazin-3(2H)-one

Referring to the synthesis method of intermediate 1g, compound 28A was prepared as a white solid (34 mg, yield 64%)

¹ H NMR (400MHz,DMSO-d ₆ )δ12.46(s,1H),8.18(d,2H),7.92(s,1H),6.29(s,1H),4.90–4.57(m,2H), 4.26(t,1H),4.24–4.11(m,2H),3.65(t,2H),3.43(t,2H),3.44–3.40(m,1H),3.20–3.16(m,1H),3.12– 3.01(m,1H),2.88–2.73(m,1H),1.84–1.66(m,1H),1.12(d,3H),δ1.10–0.94(m,3H),0.91–0.80(m,2H ),0.62–0.55(m,2H).

LC-MS m/z (ESI) = 630.70 [M+1].

Step two:

5-(((2S)-1-((4-(4-(5-cyclopropylpyrimidin-2-yl)piperazin-1-yl)-4-oxobutan-2-yl)oxy) Propan-2-yl)amino)-4-(trifluoromethyl)pyridazin-3(2H)-one (compound 28)

5-(((2S)-1-((4-(4-(5-cyclopropylpyrimidin-2-yl)piperazin-1-yl)-4-oxobutan-2-yl)oxy)propan-2-yl)amino )-4-(trifluoromethyl)pyridazin-3(2H)-one

Referring to the synthetic method of compound 15, compound 28 was prepared as a white solid (22 mg, 80% yield)

¹ H NMR (400MHz,DMSO-d ₆ )δ12.43(s,1H),8.26(s,1H),7.65(s,1H),7.58(s,1H),6.24(dd,1H),4.33– 4.26(m,2H),3.74–3.63(m,4H),3.52(d,2H),3.55–3.46(m,4H),2.59–2.55(m,2H),1.99-1.90(m,1H), 1.15(d,3H), 0.96–0.84(m,2H), 0.79–0.63(m,2H).

LC-MS m/z (ESI) = 510.20 [M+1].

Example 29

(S)-5-((1-(3-oxo-3-(4-(5-(2,2,3,3-tetrafluorocyclopropyl)pyrimidin-2-yl)piperazine-1- Base) propoxy) propan-2-yl) amino)-4-(trifluoromethyl)pyridazin-3(2H)-one (compound 29)

(S)-5-((1-(3-oxo-3-(4-(5-(2,2,3,3-tetrafluorocyclopropyl)pyrimidin-2-yl)piperazin-1-yl)propoxy)propan- 2-yl)amino)-4-(trifluoromethyl)pyridazin-3(2H)-one

Referring to the synthetic method of compound 1, compound 29 was prepared as a white solid (66 mg, yield 43%).

¹ H NMR (400MHz,DMSO-d ₆ )δ12.45(s,1H),8.65(s,1H),7.91(s,1H),6.30–6.26(m,1H),4.17–4.12(m,1H ), 3.85–3.65(m,6H), 3.64–3.46(m,7H), 2.59(t,3H), 1.15(d,3H).

LC-MS m/z (ESI) = 568.20 [M+1].

biological test

PARP Enzyme Biochemical Test Protocol:

1. Coating: 1×histone mixture (25 μL/well) for overnight coating.

2. Blocking: add Blocking buffer 100 μL/well, block for 90 minutes.

3. Compound dilution: Dilute the compound to 8 concentrations at a ratio of 1:3, and the initial concentration is 1000nM.

4. To the 12.5 μL mixture containing 10×PARP buffer, 10×PARP Assay mixture, 5×Activated DNA, and distilled water, add 2.5 μL of different concentrations of compounds and 10 μL of enzyme (1-2ng/μL), and incubate at room temperature for 1 hour.

5. Add diluted Streptavidin-HRP and incubate for 30 minutes, then add color developing solution for color development.

6. Perform chemiluminescence readings with a microplate reader.

IC ₅₀ : refers to the compound concentration at which PARP enzyme activity is inhibited by 50%.

Table 1. Compounds have inhibitory activity on PARP-7 enzyme

Conclusion: the compound of the present invention has significant biological inhibitory activity on PARP-7 target protein.

Table 2. Selectivity of compounds for PARP-1 and PARP-7 subtypes

Conclusion: Compared with the reference substance, the compound of the present invention has stronger PARP-7 selectivity.

NCI-H1373 Cell Proliferation Inhibition Protocol

1. Inoculate NCI-H1373 cells in a 96-well cell culture plate (1500 cells/well), with a volume of 80 μL per well. Incubate overnight in a 37°C, 5% CO ₂ incubator.

2. The compound was prepared into 10 mM stock solution with DMSO, and diluted with 1640 medium to 8 concentrations (1:5), the final concentrations were 10000, 2000, 400, 80, 16, 3.2, 0.64, 0.128nM.

3. The cells without drug treatment were used as the control group, and the remaining cells were treated with different drug concentrations respectively. At the same time, culture wells without cells were set up as the blank zero-adjustment group. Two parallel wells were set up for each group, and placed in 37 ° C, 5% _CO2 incubator culture.

4. After culturing for 6 days, take out the 96-well plate from the incubator, add 100 μL Cell Titer Blue working solution to each well, shake for 2 minutes, and incubate for 10 minutes.

5. Perform chemiluminescence readings with a microplate reader.

6. Calculate cell viability = (fluorescence intensity of treatment group/fluorescence intensity of control group) × 100%, and calculate half maximal inhibitory concentration (IC ₅₀ ) by curve fitting.

IC ₅₀ : refers to the compound concentration at which cell proliferation is inhibited by 50%.

Table 3. Compounds inhibit the proliferation activity of NCI-H1373 cells

Conclusion: the compound of the present invention has a significant inhibitory effect on the proliferation of NCI-H1373 cells.

Induction of drug-metabolizing enzymes CYP450

1. Incubate adherent primary human hepatocytes (3 donors), the experimental conditions are as follows: cell density 0.7×10 ⁶ cell/mL, temperature 37.0° C., 5% CO ₂ .

2. After incubating the compound with the cells for 2-3 days, test the enzyme activity of CYP450.

3, test data processing: % relative positive control activity (%of PC)=(give compound group sample activity-blank control group sample activity)/(positive control sample activity-blank control group sample activity) * 100%

Positive control group: hepatocytes were incubated with inducers of CYP450 (CYP2B6 subtype corresponds to phenobarbital 1000 μM, CYP3A4 subtype corresponds to rifampicin 25 μM).

Blank control group: cells were incubated with medium containing the same content (v/v) of organic solvent.

Table 4. Induction experiments of compounds on metabolic enzyme CYP450

Conclusion: The compound of the present invention has no potential induction effect on the activity of metabolic enzyme CYP450, while the reference substance has obvious induction activity.

The description of the present invention has described specific implementations in detail, and those skilled in the art should recognize that the above-mentioned implementations are exemplary and cannot be construed as limitations on the present invention. Under the premise of the present invention, several improvements and modifications are made to the present invention, and the technical solutions obtained by these improvements and modifications also fall within the scope of protection of the claims of the present invention.

Claims (33)

1. A compound of formula (I), or a stereoisomer or pharmaceutically acceptable salt thereof:

   

   wherein the method comprises the steps of

   R, R ', R' are each independently H or C _1-6 An alkyl group; or alternatively

   R and R 'or R and R' form, with the atom to which they are attached, a 4-to 8-membered heterocycloalkyl group;

   R ₁ 、R ₂ each independently is H or C _1-6 An alkyl group;

   L ₁ is a bond or c=o;

   l is a bond or NH;

   C ₁ is a 6-, 7-or 8-membered heterocyclic group, preferably a 6-, 7-or 8-membered heterocyclic group containing 1, 2 or 3N atoms, more preferably

   

   

   R ₃ 、R ₄ Each independently is H, halogen or C _1-6 Alkyl, wherein the C _1-6 The alkyl group is optionally further substituted with 1 or more substituents selected from OH or CN;

   c is a 6 membered heteroaryl, preferably a 6 membered heteroaryl containing 1, 2 or 3N atoms, more preferably

   

   

   The 6 membered heteroaryl optionally being further substituted with 1 or more C selected from CN or optionally with 1 or more halogen _1-6 Substituted by alkyl;

   C ₂ is C _3-6 A carbocycle;

   R ₅ each independently is H, halogen, CN or OH;

   n is 1 or 2;

   m is 0, 1, 2, 3, 4 or 5.
2. The compound of claim 1, or a stereoisomer or pharmaceutically acceptable salt thereof, wherein the heterocycloalkyl is a 4-, 5-, or 6-membered heterocycloalkyl containing 1N atom.
3. The compound of claim 1, or a stereoisomer or pharmaceutically acceptable salt thereof, wherein R is H.
4. The compound of claim 1, or a stereoisomer or pharmaceutically acceptable salt thereof, wherein R', R "are each independently H or C _1-6 An alkyl group;
5. the compound of claim 1, or a stereoisomer or pharmaceutically acceptable salt thereof, wherein R ₁ And R is ₂ H.
6. The compound of claim 1, or a stereoisomer or pharmaceutically acceptable salt thereof, wherein L ₁ C=o.
7. The compound of claim 1, or a stereoisomer or pharmaceutically acceptable salt thereof, wherein L is a bond.
8. The compound of claim 1, or a stereoisomer or pharmaceutically acceptable salt thereof, wherein L ₁ C=o, L is NH.
9. The compound of claim 1, or a stereoisomer or pharmaceutically acceptable salt thereof, wherein L ₁ C=o, L is a bond.
10. The compound of claim 1, or a stereoisomer or pharmaceutically acceptable salt thereof, wherein C ₁ Is that

    
11. The compound of claim 1, or a stereoisomer or pharmaceutically acceptable salt thereof, wherein R ₃ 、R ₄ Each independently is halogen or C _1-6 Alkyl, wherein the C _1-6 The alkyl group is optionally further substituted with 1 or more OH groups.
12. The compound of claim 1, or a stereoisomer or pharmaceutically acceptable salt thereof, wherein C is

    
13. The compound of claim 1, or a stereoisomer or pharmaceutically acceptable salt thereof, wherein C ₂ Is C ₃ Carbocycles.
14. The compound of claim 1, or a stereoisomer or pharmaceutically acceptable salt thereof, wherein R ₅ Is halogen.
15. The compound of claim 1, or a stereoisomer or pharmaceutically acceptable salt thereof, wherein n is 2.
16. The compound of claim 1, or a stereoisomer or pharmaceutically acceptable salt thereof, wherein m is 0.
17. A compound of formula (II), or a stereoisomer or pharmaceutically acceptable salt thereof:

    

    wherein the method comprises the steps of

    R ₁ 、R ₂ Each independently selected from H or C _1-6 An alkyl group;

    R ₃ 、R ₄ each independently selected from H, halogen or C _1-6 Alkyl, wherein the C _1-6 The alkyl group is optionally further substituted with 1 or more substituents selected from OH or CN;

    R ₅ each independently selected from H, halogen, CN or OH;

    l is a bond or-NH-;

    C ₁ is that

    

    C ₂ Is C _3-5 A carbocycle;

    n is 1 or 2;

    m is 0, 1, 2, 3, 4 or 5.
18. The compound of claim 1, or a stereoisomer or pharmaceutically acceptable salt thereof, wherein R ₁ And R is ₂ H.
19. The compound of claim 1, or a stereoisomer or pharmaceutically acceptable salt thereof, wherein L is a bond.
20. The compound of claim 1, or a stereoisomer or pharmaceutically acceptable salt thereof, wherein R ₃ 、R ₄ Each independently is halogen or C _1-6 Alkyl, wherein the C _1-6 The alkyl group is optionally further substituted with 1 or more OH groups.
21. The compound of claim 1, or a stereoisomer or pharmaceutically acceptable salt thereof, wherein C ₂ Is C ₃ Carbocycles.
22. The compound of claim 1, or a stereoisomer or pharmaceutically acceptable salt thereof, wherein R ₅ Is halogen.
23. The compound of claim 1, or a stereoisomer or pharmaceutically acceptable salt thereof, wherein n is 2.
24. The compound of claim 1, or a stereoisomer or pharmaceutically acceptable salt thereof, wherein m is 0.
25. A compound, or a stereoisomer or pharmaceutically acceptable salt thereof, selected from the following structures:

    

    

    
26. an intermediate compound for preparing a compound of general formula (I), (II) or a stereoisomer or a pharmaceutically acceptable salt thereof, having a compound of general formula (III), (IV), (V) or (VI):

    

    Wherein the method comprises the steps of

    R, R ', R' are each independently H or C _1-6 An alkyl group; or alternatively

    R and R 'or R and R' form, with the atom to which they are attached, a 4-to 8-membered heterocycloalkyl group;

    R ₁ 、R ₂ each independently is H or C _1-6 An alkyl group;

    L ₁ is a bond or c=o;

    l is a bond or NH;

    C ₁ is a 6-to 8-membered heterocyclic group, preferably

    

    

    R ₃ 、R ₄ Each independently is H, halogen or C _1-6 Alkyl, wherein the C _1-6 The alkyl group is optionally further substituted with 1 or more substituents selected from OH or CN;

    c is a 6 membered heteroaryl, preferably

    

    The 6 membered heteroaryl is optionally further substituted with 1 or more groups selected from CN or C _1-6 Substituted by substituents of alkyl groups, said C _1-6 Alkyl is optionally further substituted with 1 or more halogens;

    C ₂ is C _3-6 A carbocycle;

    R ₅ each independently is H, halogen, CN or OH;

    C ₃ is a 6-to 8-membered heterocyclic group, preferably

    

    

    X ₁ Is H or NH ₂ ；

    n is 1 or 2;

    m is 0, 1, 2, 3, 4 or 5.
27. The compound of claim 26, or a stereoisomer or pharmaceutically acceptable salt thereof, wherein the heterocycloalkyl is a 4-, 5-, or 6-membered heterocycloalkyl containing 1N atom.
28. The compound of claim 26, or a stereoisomer or pharmaceutically acceptable salt thereof, wherein C ₁ Is a 6-, 7-or 8-membered heterocyclic group containing 1, 2 or 3N atoms.
29. The compound of claim 26, or a stereoisomer or pharmaceutically acceptable salt thereof, wherein C is a 6 membered heteroaryl comprising 1, 2 or 3N atoms.
30. An intermediate compound according to claim 26, or a stereoisomer or pharmaceutically acceptable salt thereof, which is of the structure:

    

    
31. a pharmaceutical composition comprising:

    (1) A compound of any one of claims 1 to 25, or a stereoisomer or pharmaceutically acceptable salt thereof;

    (2) Optionally one or more other active ingredients; and

    (3) Pharmaceutically acceptable carriers and/or excipients.
32. Use of a compound according to any one of claims 1 to 25, or a stereoisomer or pharmaceutically acceptable salt thereof, or a pharmaceutical composition according to claim 31, in the manufacture of an anti-tumour medicament; preferably, the tumor formation is related to PARP; more preferably, the PARP is PARP-7.
33. Use of a compound according to any one of claims 1 to 25, or a stereoisomer or pharmaceutically acceptable salt thereof, or a pharmaceutical composition according to claim 31, for the preparation of a PARP inhibitor; preferably, the PARP is PARP-7.