TW202246261A - Compounds as anticancer agents - Google Patents

Abstract

The present invention is directed to compounds, pharmaceutically acceptable compositions comprising compounds of the invention and methods of using said compositions in the treatment of various disorders.

Description

Compounds as Anticancer Agents

The present invention relates to novel compounds (formulas I, II, III and IV) as covalent inhibitors of KRAS G12C, their synthesis and their use for the treatment of diseases.

RAS is one of the best known oncogenes. In humans, three RAS genes (HRAS, KRAS, and NRAS) encode four highly homologous RAS proteins (HRAS, KRAS-4A, KRAS-4B, and NRAS). RAS proteins are small GTPases that function as binary molecular switches involved in the transduction of extracellular growth and differentiation signals.

RAS normally cycles between a GDP-bound "off" state and a GTP-bound "on" state. This cycle is regulated by several factors. Guanine nucleotide exchange factors (GEFs), including SOS1 and SOS2, promote the exchange and formation of GTP-bound RAS. GTPase-activating proteins (GAPs), such as NF-1, promote the hydrolysis of GTP, thus switching RAS back to the GDP-bound inactive state (Kessler et al., PNAS, 2019, 116 (32): 15823-15829). Once bound to GTP, RAS initiates conformational changes in two specific regions, switch 1 and switch 2, which allow the engagement and initiation of downstream effector proteins to initiate intracellular signaling pathway cascades. These effectors include the RAF–MEK–ERK and PI3K-AKT-mTOR pathways, both of which have critical roles in regulating cell proliferation, differentiation and survival (Cox et al., Nature Reviews Drug Discovery, 2014, 13:828- 851).

RAS mutations have been identified in approximately 30% of human tumors. These mutations frequently occur as single-base missense mutations in codons 12, 13, or 61, resulting in stabilization of the initiated GTP-bound RAS form and constitutive initiation of RAS downstream signaling pathways. KRAS is the most frequently mutated RAS in cancer, accounting for 85% of all RAS-driven cancers, followed by NRAS (12%) and HRAS (3%). KRAS mutations have been detected in approximately 95% of pancreatic ductal adenocarcinomas, 50% of colorectal adenocarcinomas, and 30% of lung adenocarcinomas. Most KRAS mutations occur at residue 12, and the type of mutation varies in different cancers. In colon and pancreatic cancers, the predominant KRAS mutation is G12D (glycine to lysine), while in non-small cell lung cancer (NSCLC), nearly half of KRAS mutations are G12C (glycine to cysteamine acid) (Cox et al., Nature Reviews Drug Discovery, 2014, 13:828-851).

Based on the critical role of RAS in cell proliferation and its high mutation rate in human cancers, RAS has long been considered as a therapeutic target for many cancers. However, despite decades of research efforts, no anti-RAS small molecule has been clinically approved. The main reason is the lack of drug-targetable pockets on the RAS surface (Papke et al., Science, 2017, 355: 1158–1163). Recently, more and more studies have shown that RAS may be able to be targeted by small molecule drugs. Several inhibitors directly targeting KRAS G12C are under investigation (Patricelli et al., Cancer Discovery, 2016, 6(3); 316–29) (Fell et al., ACS Med. Chem. Lett. 2018, 9, 12, 1230-1234).

Therefore, there remains a need to provide new compounds useful as KRAS G12C inhibitors. The compounds of the present invention help to meet this need.

(I) 其中R ₁、R ₂、R ₃和R ₄各自獨立地選自氫、烷基和鹵素， 其中R ₅選自-NH ₂和-OH，並且R ₄和R ₅可以與它們所附接的基團一起形成五元或六元環， 其中X和Y各自獨立地選自N和C， 其中L選自H、N、O、SO ₂和S， 其中Q選自烷基和任選經取代的含氮五元環，條件是當L是H時，Q不存在， 其中

選自含有1或2個選自N的雜原子的6元雜芳基、或含氮雙環，它們各自任選地被-C _1-3烷基、鹵素、-CN、3,4,5,6-元碳環取代， 其中W選自-CN、任選經取代的2-丙烯-1-酮、或任選經取代的戊-2-烯-1,4-二酮，所述任選的取代基選自鹵素、烷基、-CN、3元碳環。 Aspect 1: A compound of formula I, a pharmaceutically acceptable salt thereof, or a stereoisomer thereof is provided:

(I) wherein R ₁ , R ₂ , R ₃ and R ₄ are each independently selected from hydrogen, alkyl and halogen, wherein R ₅ is selected from -NH ₂ and -OH, and R ₄ and R ₅ can be attached to them The connected groups together form a five-membered or six-membered ring, wherein X and Y are each independently selected from N and C, wherein L is selected from H, N, O, _SO and S, wherein Q is selected from alkyl and optionally Substituted nitrogen-containing five-membered rings, with the proviso that when L is H, Q is absent, wherein

selected from 6-membered heteroaryl groups containing 1 or 2 heteroatoms selected from N, or nitrogen-containing bicyclic rings, each of which is optionally replaced by -C _1-3 alkyl, halogen, -CN, 3,4,5, 6-membered carbocyclic ring substitution, wherein W is selected from -CN, optionally substituted 2-propene-1-one, or optionally substituted pent-2-ene-1,4-dione, the optionally The substituents for are selected from halogen, alkyl, -CN, and 3-membered carbocycles.

，條件是當L是H時，Q不存在， 其中

選自

、

、

、

、

、

、

和

， 其中W選自

、

、

、

、

、

、

、

和

。 Aspect 2: The compound of formula I according to Aspect 1, a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, wherein R ₁ , R ₂ , R ₃ and R ₄ are each independently selected from hydrogen, alkane and halogen, wherein R ₅ is selected from -NH ₂ and -OH, and R ₄ and R ₅ may form a five- or six-membered ring with the group to which they are attached, wherein X and Y are each independently selected from N and C, wherein L is selected from H, N, O, _SO and S, wherein Q is selected from methyl and

, provided that when L is H, Q does not exist, where

selected from

,

,

,

,

,

,

with

, where W is selected from

,

,

,

,

,

,

,

with

.

， 其中

選自

、

、

、

、

、

、

和

， 其中W選自

、

、

、

、

、

、

、

和

。 Aspect 3: A compound of formula I according to aspect 1 or 2, a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, wherein R ₁ and R ₂ are each independently selected from halogen, R ₃ and R ₄ Each is independently selected from hydrogen and alkyl, wherein R is _-OH , wherein X and Y are each C, wherein L is selected from H, O, and S, wherein Q is selected from

, in

selected from

,

,

,

,

,

,

with

, where W is selected from

,

,

,

,

,

,

,

with

.

， 其中

選自

、

、

、

和

， 其中W選自

、

、

、

、

、

和

。 Aspect 4: A compound of _formula I according to any one of aspects 1-3, a pharmaceutically acceptable salt thereof, or _a stereoisomer thereof, wherein R and R are each independently selected from F or Cl , R ₃ and R ₄ are each independently selected from hydrogen and C _1-3 alkyl, wherein R ₅ is -OH, wherein X and Y are each C, wherein L is selected from H, O and S, wherein Q is selected from

, in

selected from

,

,

,

with

, where W is selected from

,

,

,

,

,

with

.

。 Aspect 5: The compound of formula I according to any one of aspects 1-4, a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, wherein L is selected from O and S, and Q is selected from

.

，

選自

。 Aspect 6: A compound of formula I according to any one of aspects 1-5, a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, wherein L is selected from O and S, and Q is selected from

,

selected from

.

選自

。 Aspect 7. A compound of formula I, a pharmaceutically acceptable salt thereof, or a stereoisomer thereof according to any one of aspects 1-4, wherein L is H,

selected from

.

和

。 Aspect 8: A compound of formula I according to any one of aspects 1-7, a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, wherein W is selected from

with

.

選自

，W是

。 Aspect 9: A compound of formula I, a pharmaceutically acceptable salt thereof, or a stereoisomer thereof according to any one of aspects 1-8, wherein L is H, and

selected from

, W is

.

，並且

選自

，W選自

和

。 Aspect 10: A compound of formula I according to any one of aspects 1-9, a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, wherein L is selected from O and S, and Q is selected from

,and

selected from

, W selected from

with

.

(II) 其中R各自獨立地選自氫、C _1-6烷基和鹵素， R ₂選自-NH ₂和-OH， X是C， L選自H、N、O、SO ₂和S， Q選自烷基和任選經取代的含氮五元環，條件是當L是H時，Q不存在，

選自含有1或2個選自N的雜原子的6元雜芳基、或含氮雙環，它們各自任選地被氫、-C _1-3烷基、鹵素、-CN、3,4,5,6-元碳環取代，並且 R ₁和R ₃各自獨立地選自氫、-C _1-3烷基和鹵素。 Aspect 11: A compound of formula II, a pharmaceutically acceptable salt thereof, or a stereoisomer thereof is provided:

(II) wherein each R is independently selected from _hydrogen , C _1-6 alkyl and halogen, R is selected from -NH and _-OH , X is C, L is selected from H, N, O, _SO and S, Q is selected from alkyl and optionally substituted nitrogen-containing five-membered rings, with the proviso that when L is H, Q is absent,

selected from 6-membered heteroaryl groups containing 1 or 2 heteroatoms selected from N, or nitrogen-containing bicyclic rings, each of which is optionally replaced by hydrogen, -C _1-3 alkyl, halogen, -CN, 3,4, 5,6-membered carbocyclic ring substitution, and R ₁ and R ₃ are each independently selected from hydrogen, -C _1-3 alkyl and halogen.

選自

、

、

、

和

，並且 R ₁和R ₃各自獨立地選自氫、-C _1-3烷基和鹵素。 Aspect 12: There is provided a compound of formula II according to aspect 11, a pharmaceutically acceptable salt thereof, or a stereoisomer thereof: wherein each R is independently selected from hydrogen, C _1-3 alkyl and F, R2 is _-OH , X is C, L is H,

selected from

,

,

,

with

, and R ₁ and R ₃ are each independently selected from hydrogen, -C _1-3 alkyl and halogen.

選自

和

，並且 R ₁和R ₃各自獨立地選自氫、-C _1-3烷基和鹵素。 Aspect 13: There is provided a compound of formula II according to aspect 11 or 12, a pharmaceutically acceptable salt thereof, or a stereoisomer thereof: wherein each R is independently selected from hydrogen and F, R ₂ is -OH , X is C, L is H,

selected from

with

, and R ₁ and R ₃ are each independently selected from hydrogen, -C _1-3 alkyl and halogen.

(III) 其中R各自獨立地選自氫、C _1-6烷基和鹵素， R ₂選自-NH ₂和-OH， X是C， L選自H、N、O、SO ₂和S， Q選自烷基和任選經取代的含氮五元環，條件是當L是H時，Q不存在，

選自含有1或2個選自N的雜原子的6元雜芳基、或含氮雙環，它們各自任選地被氫、-C _1-3烷基、鹵素、-CN、3,4,5,6-元碳環取代，並且 R ₁和R ₃各自獨立地選自氫、-C _1-3烷基和鹵素。 Aspect 14: A compound of formula III, a pharmaceutically acceptable salt thereof, or a stereoisomer thereof is provided:

(III) wherein R is each independently selected from _hydrogen , C _1-6 alkyl and halogen, R is selected from -NH and _-OH , X is C, L is selected from H, N, O, _SO and S, Q is selected from alkyl and optionally substituted nitrogen-containing five-membered rings, with the proviso that when L is H, Q is absent,

selected from 6-membered heteroaryl groups containing 1 or 2 heteroatoms selected from N, or nitrogen-containing bicyclic rings, each of which is optionally replaced by hydrogen, -C _1-3 alkyl, halogen, -CN, 3,4, 5,6-membered carbocyclic ring substitution, and R ₁ and R ₃ are each independently selected from hydrogen, -C _1-3 alkyl and halogen.

選自

、

、

、

和

，並且 R ₁和R ₃各自獨立地選自氫、-C _1-3烷基和F。 Aspect 15: There is provided a compound of formula III according to aspect 14, a pharmaceutically acceptable salt thereof, or a stereoisomer thereof: wherein each R is independently selected from hydrogen, C _1-3 alkyl and F, R ₂ is -OH, X is C, L is selected from H, O, SO ₂ and S, Q is selected from C _1-6 alkyl, with the proviso that when L is H, Q is absent,

selected from

,

,

,

with

, and R ₁ and R ₃ are each independently selected from hydrogen, -C _1-3 alkyl and F.

是

，並且 R ₁和R ₃各自獨立地選自氫和-C _1-3烷基。 Aspect 16: There is provided a compound of formula III according to aspect 14 or 15, a pharmaceutically acceptable salt thereof, or a stereoisomer thereof: wherein each R is independently selected from hydrogen and F, R ₂ is -OH , X is C, L is selected from H and S, Q is selected from C _1-3 alkyl, with the proviso that when L is H, Q is absent,

yes

, and R ₁ and R ₃ are each independently selected from hydrogen and -C _1-3 alkyl.

(IV) 其中R各自獨立地選自氫、C _1-6烷基和鹵素， R ₂選自-NH ₂和-OH， X是C， L選自H、N、O、SO ₂和S， Q選自烷基和任選經取代的含氮五元環，條件是當L是H時，Q不存在，

選自含有1或2個選自N的雜原子的6元雜芳基、或含氮雙環，它們各自任選地被氫、-C _1-3烷基、鹵素、-CN、3,4,5,6-元碳環取代，並且 R ₁和R ₃各自獨立地選自氫、-C _1-3烷基和鹵素。 Aspect 17: A compound of formula IV, a pharmaceutically acceptable salt thereof, or a stereoisomer thereof is provided:

(IV) wherein R is each independently selected from _hydrogen , C _1-6 alkyl and halogen, R is selected from -NH and _-OH , X is C, L is selected from H, N, O, _SO and S, Q is selected from alkyl and optionally substituted nitrogen-containing five-membered rings, with the proviso that when L is H, Q is absent,

selected from 6-membered heteroaryl groups containing 1 or 2 heteroatoms selected from N, or nitrogen-containing bicyclic rings, each of which is optionally replaced by hydrogen, -C _1-3 alkyl, halogen, -CN, 3,4, 5,6-membered carbocyclic ring substitution, and R ₁ and R ₃ are each independently selected from hydrogen, -C _1-3 alkyl and halogen.

，條件是當L是H時，Q不存在，

選自

、

、

、

和

，並且 R ₁和R ₃各自獨立地選自氫、-C _1-3烷基和F。 Aspect 18: There is provided a compound of formula III according to aspect 17, a pharmaceutically acceptable salt thereof, or a stereoisomer thereof: wherein each R is independently selected from hydrogen, C _1-3 alkyl and F, R2 is _-OH , X is C, L is selected from H, O and S, Q is

, provided that when L is H, Q does not exist,

selected from

,

,

,

with

, and R ₁ and R ₃ are each independently selected from hydrogen, -C _1-3 alkyl and F.

，條件是當L是H時，Q不存在，

選自

、

、

、

和

，並且 R ₁和R ₃各自獨立地選自氫、-C _1-3烷基和F。 Aspect 19: There is provided a compound of formula III according to aspect 17, a pharmaceutically acceptable salt thereof, or a stereoisomer thereof: wherein each R is independently selected from hydrogen, C _1-3 alkyl and F, R2 is _-OH , X is C, L is selected from H, O and S, Q is

, provided that when L is H, Q does not exist,

selected from

,

,

,

with

, and R ₁ and R ₃ are each independently selected from hydrogen, -C _1-3 alkyl and F.

。 Aspect 20: The compound according to any one of Aspects 1-19, a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, wherein said compound is selected from the group consisting of:

.

Aspect 21: A pharmaceutical composition comprising the compound according to any one of Aspects 1-20, a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, and one or more pharmaceutically acceptable carriers , diluent or excipient.

Aspect 22: A method of treating various disorders comprising administering to a subject in need thereof a compound according to any one of aspects 1-20, a pharmaceutically acceptable salt thereof, or a stereoisomer thereof.

Aspect 23: The method of Aspect 22, wherein the disorder is cancer.

Unless explicitly defined elsewhere in this document, all other technical and scientific terms used herein have the meaning commonly understood by one of ordinary skill in the art to which this invention belongs.

The following terms have indicated meanings throughout the specification:

As used herein (including the appended claims), singular words such as "a," "an," and "the" include their corresponding plural referents unless the context clearly dictates otherwise.

The term "or" is used to mean and is used interchangeably with the term "and/or" unless the context clearly dictates otherwise.

The term "alkyl" refers to the group consisting of 1 to 18 (such as 1 to 12, further such as 1 to 10, further such as 1 to 8, or 1 to 6, or 1 to 4) carbon atoms The hydrocarbon groups in the straight chain and branched saturated hydrocarbon groups. Examples of alkyl groups containing 1 to 6 carbon atoms (i.e., C _1-6 alkyl groups) include, but are not limited to, methyl, ethyl, 1-propyl or n-propyl ("n-Pr"), 2- Propyl or isopropyl (“i-Pr”), 1-butyl or n-butyl (“n-Bu”), 2-methyl-1-propyl or isobutyl (“i-Bu”) , 1-methylpropyl or secondary butyl (“s-Bu”), 1,1-dimethylethyl or tertiary butyl (“t-Bu”), 1-pentyl, 2-pentyl Base, 3-pentyl, 2-methyl-2-butyl, 3-methyl-2-butyl, 3-methyl-1-butyl, 2-methyl-1-butyl, 1-hexyl , 2-hexyl, 3-hexyl, 2-methyl-2-pentyl, 3-methyl-2-pentyl, 4-methyl-2-pentyl, 3-methyl-3-pentyl, 2 -Methyl-3-pentyl, 2,3-dimethyl-2-butyl and 3,3-dimethyl-2-butyl. Alkyl as defined herein is optionally deuterated or tritiated.

The term "propyl" refers to 1-propyl or n-propyl ("n-Pr"), 2-propyl or isopropyl ("i-Pr").

The term "butyl" refers to 1-butyl or n-butyl ("n-Bu"), 2-methyl-1-propyl or isobutyl ("i-Bu"), 1-methylpropyl Or secondary butyl (“s-Bu”), 1,1-dimethylethyl or tertiary butyl (“t-Bu”).

The term "pentyl" refers to 1-pentyl, 2-pentyl, 3-pentyl, 2-methyl-2-butyl, 3-methyl-2-butyl, 3-methyl-1-butyl base, 2-methyl-1-butyl.

The term "hexyl" refers to 1-hexyl, 2-hexyl, 3-hexyl, 2-methyl-2-pentyl, 3-methyl-2-pentyl, 4-methyl-2-pentyl, 3- Methyl-3-pentyl, 2-methyl-3-pentyl, 2,3-dimethyl-2-butyl and 3,3-dimethyl-2-butyl.

The term "halogen" refers to fluorine (F), chlorine (Cl), bromine (Br) and iodine (I).

As used herein, unless otherwise indicated, the term "aryl" refers to an unsubstituted or substituted monocyclic or polycyclic aromatic ring system containing carbon ring atoms. Preferred aryl groups are monocyclic or bicyclic 6-10 membered aromatic ring systems. Phenyl and naphthyl are preferred aryl groups. The most preferred aryl is phenyl.

The term "carbocycle" refers to a substituted or unsubstituted monocyclic, bicyclic or polycyclic non-aromatic saturated ring optionally including an alkylene linker to which a cycloalkyl group can be attached. Exemplary "cycloalkyl" groups include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, and the like.

As referred to herein, the term "substituted" means that at least one hydrogen atom is replaced by a non-hydrogen group, provided that normal valences are maintained and that the substitution results in a stable compound. As used herein, a ring double bond is a double bond formed between two adjacent ring atoms (eg, C=C, C=N or N=N).

When any variable occurs more than one time in any constituent or formula of a compound, its definition on each occurrence is independent of its definition at every other occurrence. Thus, for example, if a group is shown to be substituted with 0-3 R groups, said group may optionally be substituted with up to three R groups, and R at each occurrence is independently selected from the definition of R. Also, combinations of substituents and/or variables are permissible only if such combinations result in stable compounds.

Where a bond to a substituent is shown to cross a bond connecting two atoms in a ring, then such substituent may be bonded to any atom on the ring. When a substituent is listed without indicating the atom to which such substituent is bonded to the rest of the compound of a given formula, then such substituent may be bonded via any atom in such substituent. Combinations of substituents and/or variables are permissible only if such combinations result in stable compounds.

Throughout the specification and appended claims, a given chemical formula or name shall include all stereo and optical isomers and racemates thereof, where such isomers exist. Unless otherwise stated, all chiral (enantiomers and diastereoisomers) and racemic forms are within the scope of the invention. Many geometric isomers of C=C double bonds, C=N double bonds, ring systems, etc. may also exist in compounds, and all such stable isomers are contemplated in the present invention. Cis- and trans- (or E- and Z-) geometric isomers of the compounds of the invention are described and may be isolated as a mixture of isomers or as isolated isomeric forms. The compounds of the invention can be isolated in optically active or racemic forms. Optically active forms can be prepared by resolution of racemic forms or by synthesis from optically active starting materials. All processes used to prepare the compounds of the invention and intermediates prepared therein are considered to be part of the invention. When enantiomeric or diastereomeric products are prepared, they may be separated by conventional methods, for example by chromatography or fractional crystallization. Depending on the process conditions, the end products of the invention are obtained in free (neutral) or salt form. Both the free forms and salts of these end products are within the scope of the present invention. A compound in one form can be converted to the other if so desired. A free base or acid can be converted into a salt; a salt can be converted into the free compound or another salt; a mixture of isomeric compounds of the invention can be separated into the individual isomers. The compounds of the invention (free form and their salts) can exist in various tautomeric forms in which the hydrogen atoms are transposed to other parts of the molecule and thus the chemical bonds between the atoms of the molecule are rearranged. It is to be understood that all tautomeric forms where they can exist are included in the present invention.

The invention includes that the compounds described may contain one or more asymmetric centers and thus may give rise to diastereoisomers and optical isomers. The present invention includes all such possible diastereoisomers as well as their racemic mixtures, their substantially pure resolved enantiomers, all possible geometric isomers, and their pharmaceutically acceptable of salt.

The present invention includes all stereoisomers of the compounds and pharmaceutically acceptable salts thereof. Also included are mixtures of stereoisomers as well as isolated specific stereoisomers. During the course of the synthetic procedures used to prepare such compounds or using racemization or epimerization procedures known to those skilled in the art, the products of such procedures may be mixtures of stereoisomers.

The term "stereoisomer" as used in the present invention refers to isomers in which atoms or groups of atoms in a molecule are linked to each other in the same order but differ in spatial arrangement, including configurational isomers and configurational isomers . Configuration isomers include geometric isomers and optical isomers, and optical isomers mainly include enantiomers and diastereomers.

When a substituent is referred to as "optionally substituted", the substituent is selected from, for example, substituents such as alkyl, cycloalkyl, aryl, heterocycle, halo, hydroxy, alkoxy, pendant Oxygen, alkanoyl, aryloxy, alkanoyloxy, amine, alkylamine, arylamine, arylalkylamine, disubstituted amine (where two amino substituents selected from alkyl, aryl or arylalkyl); alkanamido, arylamido, aralkanamido, substituted alkanamido, substituted arylamino, substituted aralkyl Amide, thiol, alkylthio, arylthio, arylalkylthio, alkylthiocarbonyl, arylthiocarbonyl, arylalkylthiocarbonyl, alkylsulfonyl, arylsulfonyl, Arylalkylsulfonyl, sulfonylamino (e.g. -SO2NH2), substituted sulfonylamido, nitro, cyano, carboxyl, carbamoyl (e.g. -CONH2), substituted carbamoyl (e.g. -CONHalkyl, -CONHaryl, -CONHarylalkyl, or where there are two substituents on the nitrogen selected from alkyl, aryl, or arylalkyl); alkoxycarbonyl , aryl, substituted aryl, guanidinyl, heterocyclic groups (such as indolyl, imidazolyl, furyl, thienyl, thiazolyl, pyrrolidyl (pyrrolidyl), pyridyl, pyrimidyl, pyrrolidyl ( pyrrolidinyl), piperidinyl, morpholinyl, piperazinyl, homopiperazinyl, etc.), and substituted heterocyclyl, unless otherwise defined.

For clarity and in accordance with standard practice in the art, symbols are used in formulas and tables to show bonds, which are points of attachment of moieties or substituents to the core/core of the structure.

Also, for clarity, when a substituent has a dash (-) that is not between two letters or symbols, this is used to indicate the point of attachment of the substituent. For example, -CONH2 is attached through a carbon atom.

Also, for clarity, when no substituents are shown at the end of a solid line, this indicates the presence of a methyl (CH3) group attached to a bond.

As used herein, "pharmaceutically acceptable salts" refers to derivatives of the disclosed compounds wherein the parent compound is modified by making acid or base salts thereof. Examples of pharmaceutically acceptable salts include, but are not limited to, mineral or organic acid salts of basic groups such as amines; and basic or organic salts of acidic groups such as carboxylic acids. Pharmaceutically acceptable salts include conventional non-toxic salts or quaternary ammonium salts of the parent compound formed, for example, from non-toxic inorganic or organic acids. Such conventional nontoxic salts include, for example, those derived from inorganic acids such as hydrochloric, hydrobromic, sulfuric, sulfamic, phosphoric, and nitric acids; and salts prepared from organic acids such as acetic, propionic, succinic, Acid, glycolic acid, stearic acid, lactic acid, malic acid, tartaric acid, citric acid, ascorbic acid, pamoic acid (pamoic), maleic acid, hydroxymaleic acid, phenylacetic acid, glutamic acid, benzoic acid, salicylic acid acid, sulfanilic acid, 2-acetyloxybenzoic acid, fumaric acid, toluenesulfonic acid, methanesulfonic acid, ethanedisulfonic acid, oxalic acid, and isethionic acid, etc.

The pharmaceutically acceptable salts of the present invention can be synthesized from the parent compound containing a basic or acidic moiety by conventional chemical methods. Generally, such salts can be obtained by reacting the free acid or base form of these compounds with a stoichiometric amount of the appropriate base or acid in water or in an organic solvent or a mixture of both (usually preferably a non-aqueous medium such as diethyl ether). , Ethyl acetate, ethanol, isopropanol or acetonitrile) prepared by reaction. A list of suitable salts is found in Remington: The Science and Practice of Pharmacy, 22nd Ed., Allen, L. V. Jr., ed.; Pharmaceutical Press, London, UK (2012), the disclosure of which is hereby incorporated by reference.

For therapeutic use, the salts of the compounds of the invention are considered pharmaceutically acceptable. However, salts of acids and bases which are not pharmaceutically acceptable may also find use, for example, in the preparation or purification of a pharmaceutically acceptable compound.

therapeutic application

Disclosed herein is a method of inhibiting KRAS G12C activity comprising administering to an individual a compound disclosed herein or a pharmaceutically acceptable salt thereof, including a compound of formula (I) or a specific compound exemplified herein.

Disclosed herein is a method of treating a disease or disorder in a patient comprising administering to the patient a therapeutically effective amount of a compound disclosed herein as a KRAS G12C inhibitor, or a pharmaceutically acceptable salt thereof, wherein the compound disclosed herein comprises the formula The compound of (I) or the specific compound exemplified herein. In some embodiments, the disease or disorder is associated with inhibition of KRAS G12C interaction. Preferably, the disease or disorder is cancer.

All formulations of the pharmaceutical compositions disclosed herein can be produced by conventional methods in the field of pharmacy. For example, the active ingredient can be mixed with one or more excipients, and thereby prepare the desired formulation.

The term "disease" refers to any disease, disorder, condition, symptom or indication, and is interchangeable with the terms "disorder" or "condition".

Preparation

The compounds of the present invention can be synthesized in a variety of ways described below and well known to those skilled in the art of organic synthesis, as well as synthetic methods known in the field of synthetic organic chemistry or variations thereof understood by those skilled in the art. Preferred methods are not limited to those described below. References cited herein are all incorporated by reference in their entirety.

The synthetic methods described below are intended to illustrate the invention without limiting its subject matter and the scope of the compounds claimed for these examples. Where the preparation of starting compounds is not described, they are either commercially available or can be prepared analogously to known compounds or processes described herein. Materials described in the literature were prepared according to published synthetic methods. Compounds of formulas I-IV can be synthesized by referring to the methods illustrated in the following schemes. As shown herein, the final compound is a product having the same structural formula as depicted in Formulas I-IV. It is understood that any of the compounds of formulas I-IV may be prepared by selection of reagents with appropriate substitutions. Solvents, temperature, pressure and other reaction conditions can be readily selected by one of ordinary skill in the art. Protecting groups were manipulated according to standard methods of organic synthesis (T.W. Green and P.G. M. Wuts (1999) Protective Groups in Organic Synthesis, 3rd edition, John Wiley & Sons). These groups are removed at some stage in the compound synthesis using methods clear to those skilled in the art.

Compounds of formula I-IV can be prepared by referring to the methods illustrated in the following schemes. As shown herein, the final product is a compound having the same structural formula as Formulas I-IV. It is understood that compounds of any of formulas I-IV may be produced by appropriate selection of protocols with appropriate substitution of reagents. Solvents, temperature, pressure and other reaction conditions can be readily selected by one of ordinary skill in the art. Starting materials are either commercially available or readily prepared by one of ordinary skill in the art. The constituents of a compound are as defined herein or elsewhere in the specification.

plan 1

When L is not hydrogen:

When L is hydrogen:

Scenario 2

Option 3

Option 4

example

The invention is further defined in the following examples. It should be understood that the examples are given by way of illustration only. From the above discussion and examples, one skilled in the art can ascertain the essential characteristics of this invention, and without departing from the spirit and scope thereof, can make various changes and modifications to adapt the invention to various usages and conditions. Accordingly, the present invention is not limited to the illustrative examples set forth herein below, but is instead defined by the claims appended hereto.

The following table shows some abbreviations of the present invention: aq Water-based EtOH ethanol NCS N-chlorosuccinimide g gram DCM Dichloromethane h Hour TEA Triethylamine HPLC HPLC DMF Dimethylformamide LC-MS Liquid Chromatography-Mass Spectrometry DMSO Dimethyridine Me methane Equiv equivalent ACN Acetonitrile Et ₃ N Triethylamine min minute THF Tetrahydrofuran mL Millimoles EtOAc ethyl acetate NH ₄ Cl ammonium chloride Pd(dppf)Cl ₂ [1,1'-Bis(diphenylphosphino)ferrocene]dichloropalladium(II) PE petroleum ether Prep-TLC preparative thin layer chromatography Na ₂ SO ₄ sodium sulfate

Intermediate preparation

Starting materials for the preparation of Intermediates and Examples are commercially available unless otherwise stated.

intermediate-1

6,7-Dichloropyrido[2,3-d]pyrimidin-4-ol

Step 1: 2-Amino-5,6-dichloronicotinic acid

To a solution of 2-amino-6-chloronicotinic acid (25.0 g, 0.14 mol) in DMF (125 mL) was added NCS (20.5 g, 0.15 mol). The reaction mixture was stirred at 70°C for 12 h. The reaction solution was cooled to 25 ºC and poured into water (500 mL). Filtration, and the filter cake was washed with water (50 mL x 2), then dried to afford the title product (26.0 g, 90% yield) as a white solid. LCMS: MS m/z (ESI): 206.9 [M+H] ⁺ .

Step 2: 2-Amino-5,6-dichloronicotinamide

To a solution of 2-amino-5,6-dichloronicotinic acid (26.0 g, 0.13 mol) in THF (260 mL) was added SOCl ₂ (30 mL). The mixture was stirred overnight at room temperature. The reaction mixture was concentrated. To the residue was added THF (260 mL) and ammonia (100 mL). The mixture was stirred at room temperature for 1 h. The reaction mixture was concentrated to afford the title product (26.0 g, 100% yield) as a white solid. LCMS: MS m/z (ESI): 205.9 [M+H] ⁺ .

Step 3: 6,7-Dichloropyrido[2,3-d]pyrimidin-4-ol

A solution of 2-amino-5,6-dichloronicotinamide (6 g, 29.1 mmol) in (EtO) ₃ CH (60 mL) was heated at reflux for 4 h. The reaction mixture was filtered, washed with petroleum ether (10 mL x 3). The filter cake was filtered to afford the title product (3.7 g, 60% yield) as a yellow solid. 1H NMR (400 MHz, DMSO): 13.08 (s, 1H), 8.65 (s, 1H), 8.39 (s, 1H).

Intermediate-2

6,7-Dichloropyrido[2,3-d]pyrimidine-2,4-diol

Step 1: 6,7-Dichloropyrido[2,3-d]pyrimidine-2,4-diol

To a solution of 2-amino-5,6-dichloronicotinamide (20.0 g, 97.0 mmol) in toluene (200 mL) was added oxalyl chloride (10 mL). The mixture was stirred at 110 ºC for 1 h. The mixture was cooled to room temperature, filtered, and washed with toluene (20 mL x 2). The filter cake was dried to afford the title product (10.0 g, 45% yield) as a light yellow solid. ¹ H NMR (400 MHz, CDCl3): δ 12.05 (d, J = 1.7 Hz, 1H), 11.69 (s, 1H), 8.39 (s, 1H).

Example 1

(S)-1-(4-(8-(2-fluoro-6-hydroxyphenyl)-9-isopropyl-9H-purin-6-yl)-3-methylpiperazin-1-yl) prop-2-en-1-one

Step 1: 6-Chloro-N ⁴ -isopropylpyrimidine-4,5-diamine

A mixture of 4,6-dichloropyrimidin-5-amine (10 g, 0.061 mol), propan-2-amine (17.49 g, 0.183 mol), TEA (92 g, 0.917 mol) and dry DMSO (200 ml) Stir at 120°C for 16 h. The mixture was cooled to room temperature, poured into EtOAc (900 mL), and washed with H ₂ O (400 mL x 3). The combined organics were washed with brine (200 ml x ₃ ), dried over _Na2SO4 , filtered and concentrated. The residue was purified by SGC (PE:EA = 2:1) to give the title product (8.1 g, 71%) as a white solid. LCMS: MS m/z (ESI): 187.1 [M+H] ⁺ .

Step 2: 6-Chloro-8-(2-fluoro-6-methoxyphenyl)-9-isopropyl-9H-purine

To a solution of 6-chloro- ^N4 -isopropylpyrimidine-4,5-diamine (420 mg, 2.26 mmol) in dry DMF (20 mL) was added 2-fluoro-6-methoxybenzaldehyde ( 417 mg, 2.71 mmol) and iron(III) chloride hexahydrate (732 mg, 2.71 mmol). Heat the dark brown solution to 90ºC in an open vessel for 14 h. The reaction mixture was cooled to room temperature, poured into water (20 mL), and extracted with EtOAc (20 mL x 3). The combined organics were washed with water (20 mL x 3) and brine (20 mL x 2), dried over Na ₂ SO ₄ , filtered and concentrated. The residue was purified by SGC (PE:EA = 5:1) to give the title product (400 mg, 56%) as a yellow solid. LCMS: MS m/z (ESI): 321.1 [M+H] ⁺ .

Step 3: (S)-4-(8-(2-fluoro-6-methoxyphenyl)-9-isopropyl-9H-purin-6-yl)-3-methylpiperazine-1- Tertiary butyl formate

6-Chloro-8-(2-fluoro-6-methoxyphenyl)-9-isopropyl-9H-purine (1 g, 3 mmol), (S)-3-methylpiperazine-1 - A mixture of tert-butyl formate (2 g, 10 mmol), TEA (5 g, 50 mmol) and dry DMSO (100 mL) was stirred at 110 ºC for 16 h. The reaction was cooled to room temperature and poured into water (200 mL), and extracted with EtOAc (200 mL x 3). The combined organics were washed with water (200 mL x 3) and brine (200 mL x 2), dried over Na ₂ SO ₄ , filtered and concentrated. The residue was purified by SGC (PE:EA = 4:1) to give the title product (1.0 g, 66.7%) as a yellow solid. LCMS: MS m/z (ESI): 484.6 [M+H] ⁺ .

Step 4: (S)-3-Fluoro-2-(9-isopropyl-6-(2-methylpiperazin-1-yl)-9H-purin-8-yl)phenol

At 0ºC, to (S)-4-(8-(2-fluoro-6-methoxyphenyl)-9-isopropyl-9H-purin-6-yl)-3-methylpiperazine- To a solution of tert-butyl 1-carboxylate (500 mg, 1.03 mmol) in DCM (15 mL) was added _BBr3 (2.6 g, 10.38 mmol) dropwise. The mixture was stirred at 25 ºC for 4 h. The mixture was diluted with H ₂ O (40 mL), and extracted with DCM (60 mL). The aqueous phase was adjusted to pH = 8 with NaHCO ₃ (40 mL, sat) and extracted with DCM (80 mL). The organic layer was concentrated to afford the title product (300 mg, 79%) as a white solid. LCMS: MS m/z (ESI): 371.1 [M+H] ⁺ .

Step 5: (S)-1-(4-(8-(2-fluoro-6-hydroxyphenyl)-9-isopropyl-9H-purin-6-yl)-3-methylpiperazine-1 -yl)prop-2-en-1-one

To (S)-3-fluoro-2-(9-isopropyl-6-(2-methylpiperazin-1-yl)-9H-purin-8-yl)phenol (150 mg, 0.40 mmol) and To a solution of acrylic acid (29 mg, 0.40 mmol) in DMF (2 mL) was added DIEA (155 mg, 1.20 mmol) and HATU (304 mg, 0.80 mmol). The mixture was stirred at 25 ºC for 2 h. The mixture was diluted with H ₂ O (30 mL), and extracted with EtOAc (50 mL). The combined organic layers were concentrated, and the residue was purified by preparative TLC (DCM:MeOH=20:1) to afford crude product (60 mg). The crude product was purified by preparative HPLC to afford the title product (15.06 mg, 9%) as a white solid. LCMS: MS m/z (ESI): 371.1 [M+H]+. ¹ H NMR (400 MHz, DMSO-d ₆ ): δ 8.30 (s, 1H), 7.42 (dd, J = 15.6, 8.2 Hz, 1H), 6.89-6.80 (m, 3H), 6.17 (d, J = 16.5 Hz, 1H), 5.72 (dd, J = 10.4, 2.0 Hz, 1H), 5.55-5.19 (m, 1H), 4.45-4.40 (m, 3H), 3.57-3.42 (m, 2H), 3.10-2.90 (m, 2H), 1.54 (s, 6H), 1.16 (s, 3H).

Example 2

(S)-1-(4-(2-(2-fluoro-6-hydroxyphenyl)-2H-pyrazolo[3,4-d]pyrimidin-4-yl)-3-methylpiperazine- 1-yl)prop-2-en-1-one

Step 1: (2-Fluoro-6-methoxyphenyl)hydrazine

A solution of 2-fluoro-6-methoxyaniline (10.0 g, 70.85 mmol) in HCl (100 mL) was cooled to -5 ºC and NaNO (4.89 _g , 70.87 mmol) was added dropwise with stirring over 1 h Solution in _H2O (20 mL). The reaction was stirred for 5 min, and a solution of SnCl ₂ (29.56 g, 155.89 mmol) in HCl (40 mL) was added at -5°C. The mixture was stirred at -5 ºC for 1 h. Aqueous 4 N NaOH was added to adjust pH=14. The reaction mixture was extracted with EtOAc (1500 mL). The combined organic layers were concentrated to afford the title product (10.0 g, 85%) as a brown solid. ¹ H NMR (400 MHz, DMSO-d6) δ 6.81 – 6.69 (m, 3H), 5.47 (s, 1H), 4.12 (s, 2H), 3.80 (s, 3H).

Step 2: 4-Chloro-5-(dimethoxymethyl)-6-(2-(2-fluoro-6-methoxyphenyl)hydrazino)-2-(methylthio)pyrimidine

To (2-fluoro-6-methoxyphenyl)hydrazine (5.2 g, 33.30 mmol) and 4,6-dichloro-5-(dimethoxymethyl)-2-(methylthio)pyrimidine (8.2 g, 30.46 mmol) in THF (240 mL) was added TEA (9.2 g, 91.09 mmol) and stirred at 80 ºC for 1 h. The mixture was diluted with H ₂ O (300 mL), and extracted with EtOAc (600 mL). The combined organic layers were concentrated to afford the title product (9.9 g, 100%) as a yellow solid. LCMS: MS m/z (ESI): 389.0.

Step 3: 4-Chloro-2-(2-fluoro-6-methoxyphenyl)-6-(methylthio)-2H-pyrazolo[3,4-d]pyrimidine

4-Chloro-5-(dimethoxymethyl)-6-(2-(2-fluoro-6-methoxyphenyl)hydrazino)-2-(methylthio)pyrimidine (9.9 g, 25.52 mmol) in acetone (14 mL) and _H2O (3.5 mL) was added p-TsOH (14.6 g, 76.75 mmol). The mixture was stirred at room temperature for 1 h. The mixture was concentrated and diluted with H ₂ O (100 mL), and then extracted with EtOAc (300 mL). The combined organic layers were concentrated, and the residue was purified by SGC (PE:EA = 2:1) to afford the title product (2.98 g, 48%) as a yellow solid. LCMS: MS m/z (ESI): 325.0.

Step 4: (S)-4-(2-(2-fluoro-6-methoxyphenyl)-6-(methylthio)-2H-pyrazolo[3,4-d]pyrimidine-4 -yl)-3-methylpiperazine-1-carboxylic acid tertiary butyl ester

To 4-chloro-2-(2-fluoro-6-methoxyphenyl)-6-(methylthio)-2H-pyrazolo[3,4-d]pyrimidine (2.98 g, 9.17 mmol) And to a solution of (S)-tert-butyl 3-methylpiperazine-1-carboxylate (3.67 g, 18.34 mmol) in THF (300 mL) was added DIPEA (3.56 g, 27.51 mmol). The mixture was stirred at 80 ºC for 16 h. The mixture was diluted with H ₂ O (100 mL), and then extracted with EtOAc (200 mL). The combined organic layers were concentrated, and the residue was purified by SGC (PE:EA = 1:1) to afford the title product (4.33 g, 97%) as a yellow solid. LCMS: MS m/z (ESI): 489.1.

Step 5: (S)-4-(2-(2-fluoro-6-methoxyphenyl)-2H-pyrazolo[3,4-d]pyrimidin-4-yl)-3-methylpiper Tertiary butyl oxazine-1-carboxylate

To (S)-4-(2-(2-fluoro-6-methoxyphenyl)-6-(methylthio)-2H-pyrazolo[3,4-d]pyrimidin-4-yl )-3-Methylpiperazine-1-carboxylic acid tert-butyl ester (1.0 g, 2.18 mmol) in EtOH (50 mL) was added Raney nickel (1.1 g). The mixture was stirred at 65 ºC under _H2 atmosphere for 24 h. The mixture was filtered, and the filtrate was concentrated to afford the title product (0.86 g, 86%) as a white solid. LCMS: MS m/z (ESI): 344.1.

Step 6: (S)-3-Fluoro-2-(4-(2-methylpiperazin-1-yl)-2H-pyrazolo[3,4-d]pyrimidin-2-yl)phenol

At 0ºC, to (S)-4-(2-(2-fluoro-6-methoxyphenyl)-2H-pyrazolo[3,4-d]pyrimidin-4-yl)-3-methanol To a solution of tert-butylpiperazine-1-carboxylate (800 mg, 2.04 mmol) in DCM (20 mL) was added _BBr3 /DCM (20 mL, 20.00 mmol) dropwise. The mixture was stirred at 25 ºC for 16 h. The mixture was quenched with MeOH (40 mL) and concentrated to afford the title product (636 mg, 64%) as a yellow solid. LCMS: MS m/z (ESI): 329.1.

Step 7: (S)-1-(4-(2-(2-fluoro-6-hydroxyphenyl)-2H-pyrazolo[3,4-d]pyrimidin-4-yl)-3-methyl piperazin-1-yl)prop-2-en-1-one

To (S)-3-fluoro-2-(4-(2-methylpiperazin-1-yl)-2H-pyrazolo[3,4-d]pyrimidin-2-yl)phenol (600 mg, 1.83 mmol) and acrylic acid (158 mg, 2.19 mmol) in DMF (5 mL) were added DIEA (2.36 g, 18.28 mmol) and HATU (1.39 g, 3.66 mmol). The mixture was stirred at 25 ºC for 2 h. The mixture was diluted with H ₂ O (50 mL), and extracted with EtOAc (150 mL). The combined organic layers were concentrated, and the residue was purified by prep-TLC (DCM/MeOH = 15/1) to afford crude product (140 mg). The crude product was purified by preparative HPLC to afford the title product (43.2 mg, 6%) as a white solid. LCMS: MS m/z (ESI): 383.0. ¹ H NMR (400 MHz, DMSO-d ₆ ): δ 8.95 (s, 1H), 7.32 (s, 1H), 7.47 – 7.39 (m, 1H), 6.95 – 6.80 (m, 3H), 6.18 (d, J = 16.8 Hz, 1H), 5.75 (dd, J = 10.4, 2.4 Hz, 1H), 5.22 – 4.67 (m, 2H), 4.59 – 3.70 (m, 3H ), 3.59 – 3.57 (m, 1H), 3.10 – 3.02 (m, 1H), 1.19 (d, J = 6.6 Hz, 3H).

Example 3

1-((S)-4-(2-(2-fluoro-6-hydroxyphenyl)-6-(((S)-1-methylpyrrolidin-2-yl)methoxy)-2H- Pyrazolo[3,4-d]pyrimidin-4-yl)-3-methylpiperazin-1-yl)prop-2-en-1-one

Step 1: 2-(4-Chloro-6-(methylthio)-2H-pyrazolo[3,4-d]pyrimidin-2-yl)-3-fluorophenol

At 0ºC, 4-chloro-2-(2-fluoro-6-methoxyphenyl)-6-(methylthio)-2H-pyrazolo[3,4-d]pyrimidine (2.0 g , 6.17 mmol) in DCM (40 mL) was added dropwise with BCl ₃ /DCM (15 mL, 15.00 mmol). The mixture was stirred at 0 to 25 ºC for 6 h. The mixture was concentrated to give a residue. The residue was diluted with H ₂ O (80 mL), and extracted with DCM (200 mL). The combined organic layers were concentrated to afford the title product (1.8 g, 95%) as a yellow solid. LCMS: MS m/z (ESI): 310.9.

Step 2: (S)-4-(2-(2-fluoro-6-hydroxyphenyl)-6-(methylthio)-2H-pyrazolo[3,4-d]pyrimidin-4-yl )-3-Methylpiperazine-1-carboxylic acid tertiary butyl ester

To 2-(4-chloro-6-(methylthio)-2H-pyrazolo[3,4-d]pyrimidin-2-yl)-3-fluorophenol (2.0 g, 6.45 mmol) and (S )-3-Methylpiperazine-1-carboxylic acid tert-butyl ester (2.58 g, 12.88 mmol) in DMSO (20 mL) was added DIPEA (2.50 g, 19.34 mmol). The mixture was stirred at 120 ºC for 2 h. The mixture was diluted with H ₂ O (100 mL), and then extracted with EtOAc (300 mL). The combined organic layers were concentrated to afford the title product (2.0 g, 66%) as a brown oil. LCMS: MS m/z (ESI): 475.1.

Step 3: (S)-4-(2-(2-fluoro-6-hydroxyphenyl)-6-(methylsulfonyl)-2H-pyrazolo[3,4-d]pyrimidine-4- Base)-3-Methylpiperazine-1-carboxylic acid tertiary butyl ester

To (S)-4-(2-(2-fluoro-6-hydroxyphenyl)-6-(methylthio)-2H-pyrazolo[3,4-d]pyrimidin-4-yl)- To a solution of tert-butyl 3-methylpiperazine-1-carboxylate (2.0 g, 4.21 mmol) in THF (40 mL) and H ₂ O (40 mL) was added H ₃ K ₅ O ₁₈ S ₄ (7.6 g, 12.36 mmol). The mixture was stirred at room temperature for 16 h. The mixture was quenched with Na ₂ SO ₃ (sat, 100 mL), diluted with H ₂ O (40 mL), and then extracted with EtOAc (300 mL). The combined organic layers were concentrated, and the residue was purified by SGC to afford the title product (2.13 g, 100%) as a white solid. LCMS: MS m/z (ESI): 507.0.

Step 4: (S)-4-(2-(2-Fluoro-6-hydroxyphenyl)-6-(((S)-1-methylpyrrolidin-2-yl)methoxy)-2H- Pyrazolo[3,4-d]pyrimidin-4-yl)-3-methylpiperazine-1-carboxylic acid tertiary butyl ester

To a solution of (S)-(1-methylpyrrolidin- ₂ -yl)methanol (0.97 g, 8.42 mmol) in THF (40 mL) was added NaH (0.842 g, 21.05 mmol). The mixture was stirred at 0°C for 20 min, and then (S)-4-(2-(2-fluoro-6-hydroxyphenyl)-6-(methylsulfonyl)-2H-pyrazolo[3, 4-d] A solution of pyrimidin-4-yl)-3-methylpiperazine-1-carboxylic acid tert-butyl ester (2.13 g, 4.21 mmol) in THF (10 mL). The mixture was stirred at 0 ºC for 20 min. The mixture was quenched with NH ₄ Cl (sat, 40 mL), diluted with H ₂ O (40 mL), and then extracted with EtOAc (200 mL). The combined organic layers were concentrated and the residue was purified by SGC to afford the title product (2.1 g, 92%) as a white solid. LCMS: MS m/z (ESI): 542.4.

Step 5: 3-Fluoro-2-(4-((S)-2-methylpiperazin-1-yl)-6-(((S)-1-methylpyrrolidin-2-yl)methoxy base)-2H-pyrazolo[3,4-d]pyrimidin-2-yl)phenol

At 0ºC, to (S)-4-(2-(2-fluoro-6-hydroxyphenyl)-6-(((S)-1-methylpyrrolidin-2-yl)methoxy)-2H -Pyrazolo[3,4-d]pyrimidin-4-yl)-3-methylpiperazine-1-carboxylic acid tert-butyl ester (800 mg, 1.48 mmol) in DCM (10 mL) was added TFA (5 mL). The mixture was stirred at room temperature for 0.5 h. The mixture was concentrated to afford the title product (620 mg, 95%) as a brown oil. LCMS: MS m/z (ESI): 442.3.

Step 6: 1-((S)-4-(2-(2-fluoro-6-hydroxyphenyl)-6-(((S)-1-methylpyrrolidin-2-yl)methoxy) -2H-pyrazolo[3,4-d]pyrimidin-4-yl)-3-methylpiperazin-1-yl)prop-2-en-1-one

At 0ºC, to 3-fluoro-2-(4-((S)-2-methylpiperazin-1-yl)-6-(((S)-1-methylpyrrolidin-2-yl) Methoxy)-2H-pyrazolo[3,4-d]pyrimidin-2-yl)phenol (620 mg, 1.41 mmol) and acrylic acid (122 mg, 1.69 mmol) in DMF (10 mL) DIEA (1.82 g, 14.08 mmol) and HATU (1.07 g, 2.82 mmol) were added. The mixture was stirred at room temperature for 30 min. The mixture was diluted with H ₂ O (50 mL), and extracted with EtOAc (150 mL). The combined organic layers were concentrated, and the residue was purified by preparative HPLC to afford the title product (8.1 mg, 1%) as a white solid. LCMS: MS m/z (ESI): 496.4. ¹ H NMR (400 MHz, DMSO-d ₆ ): δ 8.82 (s, 1H), 8.26 (s, 1H), 7.39 (q, J = 8.0 Hz, 1H), 6.96 – 6.76 (m, 3H), 6.18 (d, J = 15.2 Hz, 1H), 5.75 (dd, J = 10.6, 2.2 Hz, 1H), 5.17 – 4.33 (m, 2H), 4.43 – 3.83 (m, 7H), 3.02 – 2.94 (m, 1H ), 2.61 – 2.54 (m, 1H), 2.37 (s, 3H), 2.19 (q, J = 8.6 Hz, 1H), 1.99 – 1.91 (m, 1H), 1.74 – 1.56 (m, 3H), 1.18 ( d, J = 6.4 Hz, 3H).

Example 4

(S)-4-(4-acryloyl-2-methylpiperazin-1-yl)-6-(2-fluoro-6-hydroxyphenyl)-5,6-dihydro-7H-pyrrolo [3,4-d]pyrimidin-7-one

Step 1: N-((4,6-dichloro-2-(methylthio)pyrimidin-5-yl)methyl)-2-fluoro-6-methoxyaniline

To 4,6-dichloro-2-(methylthio)pyrimidine-5-carbaldehyde (10.0 g, 44.8 mmol) and 2-fluoro-6-methoxyaniline (7.59 g, 53.76 mmol) in DCM (300 mL) was added AcOH (2.69 g, 44.8 mmol). The mixture was stirred at 25°C for 1 h, then cooled to 0°C, and NaBH(OAc) ₃ (35.13 g, 165.76 mmol) was added portionwise. The mixture was stirred at 25 ºC for 16 h. The mixture was quenched with NH ₄ Cl (300 mL, saturated aq), diluted with H ₂ O (100 mL), and extracted with EtOAc (800 mL). The combined organic layers were concentrated, and the residue was purified by SGC (PE/EA = 20/1 ) to afford the title product (7.7 g, 50%) as a yellow solid. LCMS: MS m/z (ESI): 348.0 [M+H] ⁺ .

Step 2: (S)-4-(6-Chloro-5-(((2-fluoro-6-methoxyphenyl)amino)methyl)-2-(methylthio)pyrimidine-4- Base)-3-Methylpiperazine-1-carboxylic acid tertiary butyl ester

To N-((4,6-dichloro-2-(methylthio)pyrimidin-5-yl)methyl)-2-fluoro-6-methoxyaniline (13.3 g, 38.19 mmol) and (S )-3-Methylpiperazine-1-carboxylic acid tert-butyl ester (15.3 g, 76.40 mmol) in DMSO (150 mL) was added TEA (3.86 g, 38.15 mmol). The mixture was stirred at 120 ºC for 16 h. The mixture was cooled to room temperature. The mixture was diluted with H ₂ O (1500 mL), and extracted with EtOAc (2000 mL). The combined organic layers were concentrated, and the residue was purified by SGC (PE:EA = 20:1 ) to afford the title product (18.37 g, 94%) as a yellow solid. LCMS: MS m/z (ESI): 512.1 [M+H] ⁺ .

Step 3: (S)-4-(6-(2-fluoro-6-methoxyphenyl)-2-(methylthio)-7-oxo-6,7-dihydro-5H- Pyrrolo[3,4-d]pyrimidin-4-yl)-3-methylpiperazine-1-carboxylic acid tertiary butyl ester

(S)-4-(6-chloro-5-(((2-fluoro-6-methoxyphenyl)amino)methyl)-2-(methylthio)pyrimidin-4-yl) -3-Methylpiperazine-1-carboxylic acid tert-butyl ester (2.13 g, 4.15 mmol), Pd(dppf)Cl ₂ (339 mg, 0.42 mmol) and NaOAc (681 mg, 8.3 mmol) in EtOH (65 mL ) was stirred at 80 ºC under CO atmosphere for 16 h. The mixture was concentrated, and the residue was purified by SGC (PE:EA = 1:2) to afford the title product (2.14 g, 83%) as a yellow solid. LCMS: MS m/z (ESI): 504.1 [M+H] ⁺ .

Step 4: (S)-4-(6-(2-fluoro-6-methoxyphenyl)-7-oxo-6,7-dihydro-5H-pyrrolo[3,4-d] Pyrimidin-4-yl)-3-methylpiperazine-1-carboxylic acid tertiary butyl ester

To (S)-4-(6-(2-fluoro-6-methoxyphenyl)-2-(methylthio)-7-oxo-6,7-dihydro-5H-pyrrolo [3,4-d]Pyrimidin-4-yl)-3-methylpiperazine-1-carboxylic acid tert-butyl ester (1.1 g, 2.18 mmol) in EtOH (50 mL) was added Raney nickel ( 1.1g). The mixture was stirred at 65 ºC under _H2 atmosphere for 24 h. The mixture was filtered, and the filtrate was concentrated to afford the title product (0.86 g, 86%) as a white solid. LCMS: MS m/z (ESI): 458.2 [M+H] ⁺ .

Step 5: 6-(2-fluoro-6-hydroxyphenyl)-4-((3R)-3-methylpiperidin-4-yl)-5,6-dihydro-7H-pyrrolo[3, 4-d]pyrimidin-7-one

At 0ºC, to (S)-4-(6-(2-fluoro-6-methoxyphenyl)-7-oxo-6,7-dihydro-5H-pyrrolo[3,4- d] To a solution of pyrimidin-4-yl)-3-methylpiperazine-1-carboxylic acid tert-butyl ester (200 mg, 0.44 mmol) in DCM (5 mL) was added dropwise BBr ₃ (490 mg, 1.96 mmol). The mixture was stirred at 25 ºC for 4 h. The mixture was quenched with MeOH (20 mL) and concentrated to afford the title product (100 mg, 66%) as a yellow solid. LCMS: MS m/z (ESI): 344.1 [M+H] ⁺ .

Step 6: (S)-4-(4-Acryl-2-methylpiperazin-1-yl)-6-(2-fluoro-6-hydroxyphenyl)-5,6-dihydro-7H -pyrrolo[3,4-d]pyrimidin-7-one

To (S)-4-(6-(2-fluoro-6-methoxyphenyl)-7-oxo-6,7-dihydro-5H-pyrrolo[3,4-d]pyrimidine- To a solution of tert-butyl 4-yl)-3-methylpiperazine-1-carboxylate (70 mg, 0.20 mmol) and acrylic acid (15 mg, 76.40 mmol) in DMF (2 mL) was added DIEA (129 mg , 1.00 mmol) and HATU (152 mg, 0.40 mmol). The mixture was stirred at 25 ºC for 2 h. The mixture was diluted with H2O (30 mL), and extracted with EtOAc (50 mL). The combined organic layers were concentrated, and the residue was purified by prep-TLC and prep-HPLC to afford the title product (18.88 mg, 24%) as a white solid. LCMS: MS m/z (ESI): 398.1 [M+H]+. ¹ H NMR (400 MHz, DMSO-d ₆ ): δ 8.69 (s, 1H), 7.28 (td, J = 8.4, 6.8 Hz, 1H), 6.85 – 6.77 (m, 3H), 6.17 (d, J = 16.6 Hz, 1H), 5.73 (dd, J = 10.4, 2.4 Hz, 1H), 4.96 (dd, J = 38.8, 16.8 Hz, 2H), 4.72 (brs, 1H), 4.36 – 3.95 (m, 3H), 3.52 – 3.49 (m, 2H), 3.14 – 2.91 (m, 1H), 1.14 (d, J = 5.8 Hz, 3H).

Example 5

(S)-4-(4-acryl-2-methylpiperazin-1-yl)-6-(2-fluoro-6-hydroxyphenyl)-2-(methylsulfonyl)-5 ,6-Dihydro-7H-pyrrolo[3,4-d]pyrimidin-7-one

Step 1: (S)-4-(6-(2-fluoro-6-methoxyphenyl)-2-(methylsulfonyl)-7-oxo-6,7-dihydro-5H -Pyrrolo[3,4-d]pyrimidin-4-yl)-3-methylpiperazine-1-carboxylic acid tertiary butyl ester

To (S)-4-(6-(2-fluoro-6-methoxyphenyl)-2-(methylthio)-7-oxo-6,7-dihydro-5H-pyrrolo [3,4-d]pyrimidin-4-yl)-3-methylpiperazine-1-carboxylic acid tert-butyl ester (2.0 g, 3.98 mmol) in THF (20 mL) and _H2O (10 mL) A solution of H ₃ K ₅ O ₁₈ S ₄ (7.33 g, 11.93 mmol) was added. The mixture was stirred at room temperature for 1 h. The mixture was quenched with Na ₂ SO ₃ (sat, 100 mL), diluted with H ₂ O (40 mL), and then extracted with EtOAc (300 mL). The combined organic layers were concentrated, and the residue was purified by SGC to afford the title product (2.04 g, 98%) as a white solid. LCMS: MS m/z (ESI): 536.1

Step 2: (S)-6-(2-Fluoro-6-hydroxyphenyl)-4-(2-methylpiperazin-1-yl)-2-(methylsulfonyl)-5,6- Dihydro-7H-pyrrolo[3,4-d]pyrimidin-7-one

(S)-4-(6-(2-fluoro-6-methoxyphenyl)-2-(methylsulfonyl)-7-oxo-6,7-dihydro- A solution of tert-butyl 5H-pyrrolo[3,4-d]pyrimidin-4-yl)-3-methylpiperazine-1-carboxylate (2.0 g, 3.74 mmol) in DCM (20 mL) was gradually Add _PBr3 (5.51 g, 21.99 mmol) dropwise. The mixture was stirred at 25 ºC for 4 h. The mixture was concentrated to afford the title product (1.5 g, 96%) as a brown oil. LCMS: MS m/z (ESI): 422.0.

Step 3: (S)-4-(4-Acryl-2-methylpiperazin-1-yl)-6-(2-fluoro-6-hydroxyphenyl)-2-(methylsulfonyl )-5,6-dihydro-7H-pyrrolo[3,4-d]pyrimidin-7-one

At -70ºC, the compound (S)-6-(2-fluoro-6-hydroxyphenyl)-4-(2-methylpiperazin-1-yl)-2-(methylsulfonyl)- To a solution of 5,6-dihydro-7H-pyrrolo[3,4-d]pyrimidin-7-one (100 mg, 0.237 mmol) and DIPEA (61 mg, 0.474 mmol) in DCM (2.0 mL) was added Acryloyl chloride (17 mg, 0.189 mmol). The mixture was stirred at -70 ºC for 10 min. The reaction mixture was diluted with H ₂ O (10 mL), and extracted with EtOAc (10 mL x 3). The combined organic layers were concentrated, and the residue was purified by preparative HPLC to afford the title product (13.2 mg, 12% yield) as a white solid. LCMS: MS m/z (ESI): 476.2 [M+H] ⁺ . ¹ H NMR (400 MHz, CD ₃ OD): δ 7.31 - 7.26 (m, 1H), 6.93 - 6.67 (m, 3H), 6.28 (d, J = 16.2 Hz, 1H), 5.80 (dd, J = 10.6 , 1.8 Hz, 1H), 5.19 - 5.05 (m, 2H), 4.70 - 4.35 (m, 2H), 4.22 - 4.01 (m, 1H), 3.66 - 3.47 (m, 2H), 3.38 (s, 3H), 3.29 - 3.10 (m, 2H), 1.31 (d, J = 6.8 Hz, 3H).

Example 6

(S)-4-(6-(2-fluoro-6-hydroxyphenyl)-2-(methylsulfonyl)-7-oxo-6,7-dihydro-5H-pyrrolo[3 ,4-d]pyrimidin-4-yl)-3-methylpiperazine-1-carbonitrile

Step 1: (S)-4-(6-(2-fluoro-6-hydroxyphenyl)-2-(methylsulfonyl)-7-oxo-6,7-dihydro-5H-pyrrole And[3,4-d]pyrimidin-4-yl)-3-methylpiperazine-1-carbonitrile

(S)-6-(2-fluoro-6-hydroxyphenyl)-4-(2-methylpiperazin-1-yl)-2-(methylsulfonyl)-5,6 - To a solution of dihydro-7H-pyrrolo[3,4-d]pyrimidin-7-one (100 mg, 0.237 mmol) and NaHCO ₃ (47 mg, 0.474 mmol) in DMF (3 mL) was added BrCN ( 25 mg, 0.237 mmol), and the mixture was stirred at 0 ºC under N ₂ for 0.5 h. The mixture was diluted with H ₂ O (30 mL), and extracted with EtOAc (30 mL x 3). The combined organic layers were concentrated, and the residue was purified by preparative HPLC to afford the title product (1.78 mg, 1.7% yield) as a white solid. LCMS: MS m/z (ESI): 447.1 [M+H] ⁺ . ¹ H NMR (400 MHz, CD ₃ OD): δ 7.31 - 7.26 (m, 1H), 6.93 - 6.62 (m, 2H), 5.22 - 4.98 (m, 2H), 3.71 - 3.45 (m, 3H), 3.41 - 3.32 (m, 7H), 1.49 (d, J = 6.8 Hz, 3H).

Example 7

1-((3S)-4-(6-chloro-7-(2-fluoro-6-hydroxyphenyl)pyrido[2,3-d]pyrimidin-4-yl)-3-methylpiperazine- 1-yl)prop-2-en-1-one

Step 1: 4,6,7-Trichloropyrido[2,3-d]pyrimidine

To a solution of 6,7-dichloropyrido[2,3-d]pyrimidin-4-ol (100 mg, 0.46 mmol) and _POCl3 (213 mg, 1.39 mmol) in THF (5 ml) was added DIPEA (120 mg, 0.93 mmol). The mixture was stirred at 70 ºC for 1 h. The mixture was concentrated to afford the crude title product (110 mg) as a brown solid. LCMS: MS m/z (ESI): 233.9 [M+H] ⁺ .

Step 2: (S)-tert-butyl 4-(6,7-dichloropyrido[2,3-d]pyrimidin-4-yl)-3-methylpiperazine-1-carboxylate

4,6,7-trichloropyrido[2,3-d]pyrimidine (110 mg, 0.47 mmol), (S)-3-methylpiperazine-1-carboxylic acid tertiary butyl ester (73 mg, 0.56 mmol) and DIPEA (188 mg, 0.94 mmol) in NMP (5 mL) was heated at room temperature for 1 h. The mixture was diluted with H ₂ O (50 mL) and filtered. The filter cake was washed with H ₂ O (20 mL x 2), then dried to give the title product (130 mg, 69%) as a yellow solid. LCMS: MS m/z (ESI): 397.9 [M+H] ⁺ .

Step 3: (3S)-4-(6-Chloro-7-(2-fluoro-6-hydroxyphenyl)pyrido[2,3-d]pyrimidin-4-yl-3-methylpiperazine-1 -Tertiary butyl formate

(S)-4-(6,7-dichloropyrido[2,3-d]pyrimidin-4-yl)-3-methylpiperazine-1-carboxylic acid tertiary butyl ester (120 mg, 0.30 mmol ), ((2-fluoro-6-hydroxyphenyl)boronic acid (52 mg, 0.33 mmol), Pd(dppf)Cl ₂ (22 mg, 0.03 mmol), K ₂ CO ₃ (124 mg, 0.90 mmol) in 1 , a solution in 4-dioxane (5 mL) and H ₂ O (1 ml) was heated at 100 ºC for 2 h. The mixture was diluted with H ₂ O (50 mL) and extracted with EtOAc (50 mL x 3). The combined organic layers were concentrated and the residue was purified by column (DCM/MeOH = 10:1) to afford the title product (110 mg, 77%) as a yellow solid. LCMS: MS m/z (ESI): 474.0 [M+H] ⁺ .

Step 4: 2-(6-Chloro-4-((S)-2-methylpiperazin-1-yl)pyrido[2,3-d]pyrimidin-7-yl)-3-fluorophenol

(3S)-4-(6-chloro-7-(2-fluoro-6-hydroxyphenyl)pyrido[2,3-d]pyrimidin-4-yl-3-methylpiperazine-1-carboxylic acid A solution of tert-butyl ester (110 mg, 0.23 mmol) in DCM (1 mL) and TFA (0.3 mL) was stirred at room temperature for 2 h. Saturated NaHCO ₃ (30 mL) was added to the mixture and washed with (DCM : MeOH = 10:1, 30 mL x 5) extraction, the combined organics were dried over Na ₂ SO ₄ , filtered, and concentrated to give the crude title product (90 mg) as a yellow solid. LCMS: MS m/z (ESI): 373.9 [M+H] ⁺ .

Step 5: 1-((3S)-4-(6-chloro-7-(2-fluoro-6-hydroxyphenyl)pyrido[2,3-d]pyrimidin-4-yl)-3-methyl piperazin-1-yl)prop-2-en-1-one

At -78ºC, to 2-(6-chloro-4-((S)-2-methylpiperazin-1-yl)pyrido[2,3-d]pyrimidin-7-yl)-3-fluoro To a solution of phenol (50 mg, 0.134 mmol) and DIEA (35 mg, 0.268 mmol) in THF (3 mL) was added acryloyl chloride (12 mg, 0.134 mmol). The mixture was stirred at -78ºC for 10 min. The mixture was diluted with H ₂ O (10 mL), and extracted with EtOAc (20 x 3 mL). The combined organic layers were concentrated, and the residue was purified by preparative HPLC to afford the title product (1.7 mg, 3% yield) as a white solid. LCMS: MS m/z (ESI): 427.9 [M+H] ⁺ . ¹ H NMR (400 MHz, CDCl3): δ 9.69 (s, 1H), 8.86 (s, 1H), 8.29 (s, 1H), 7.37 (dd, J = 14.8, 7.8 Hz, 1H), 6.95 (d, J = 8.4 Hz, 1H), 6.76 (t, J = 9.2 Hz, 1H), 6.61 (s, 1H), 6.41 (d, J = 17.0 Hz, 1H), 5.81 (d, J = 10.6 Hz, 1H) , 4.93 - 4.47 (m, 2H), 4.29 (s, 1H), 4.05 - 3.87 (m, 1H), 3.70 - 3.50 (m, 2H), 3.21 - 3.06 (m, 1H), 1.49 (s, 3H) .

Example 8

(E)-1-((3S)-4-(6-chloro-7-(2-fluoro-6-hydroxyphenyl)pyrido[2,3-d]pyrimidin-4-yl)-3-methyl (Piperazin-1-yl)pent-2-ene-1,4-dione

Step 1: (E)-1-((3S)-4-(6-chloro-7-(2-fluoro-6-hydroxyphenyl)pyrido[2,3-d]pyrimidin-4-yl)- 3-Methylpiperazin-1-yl)pent-2-ene-1,4-dione

At 0ºC, 2-(6-chloro-4-((S)-2-methylpiperazin-1-yl)pyrido[2,3-d]pyrimidin-7-yl)-3-fluorophenol (40 mg, 0.11 mmol), DIEA (28 mg, 0.22 mmol) and (E)-4-oxopent-2-enoic acid (11 mg, 0.96 mmol) in DMF (2 mL) were added HATU (49 mg, 0.13 mmol). The mixture was stirred at 0 ºC for 20 min. The mixture was diluted with H ₂ O (10 mL), and extracted with EtOAc (20 x 3 mL). The combined organic layers were concentrated, and the residue was purified by preparative HPLC (FA) to afford the title product (7.07 mg, 14% yield) as a white solid. LCMS: MS m/z (ESI): 469.9 [M+H] ⁺ . ¹ H NMR (400 MHz, DMSO): δ 12.93 - 12.64 (m, 1H), 11.27 (s, 1H), 11.04 (d, J = 11.0 Hz, 1H), 10.82 (s, 1H), 10.07 - 9.73 ( m, 2H), 9.43 - 9.12 (m, 3H), 7.42 (s, 1H), 6.93 - 6.73 (m, 2H), 6.68 - 6.45 (m, 1H), 6.35 - 6.15 (m, 2H), 5.97 ( s, 1H), 4.88 (s, 3H), 3.83 (t, J = 6.6 Hz, 3H).

Example 9

(S)-1-(4-(7-(6-amino-4-methyl-3-(trifluoromethyl)pyridin-2-yl)-6-chloropyrido[2,3-d] Pyrimidin-4-yl)-3-methylpiperazin-1-yl)prop-2-en-1-one

Example 10

(E)-1-(7-(6-chloro-7-(2-fluoro-6-hydroxyphenyl)pyrido[2,3-d]pyrimidin-4-yl)-2,7-diazepine Spiro[3.5]non-2-yl)pent-2-ene-1,4-dione

Step 1: tertiary-butyl 7-(6,7-dichloropyrido[2,3-d]pyrimidin-4-yl)-2,7-diazaspiro-[3.5]nonane-2-carboxylate

To 4,6,7-trichloropyrido[2,3-d]pyrimidine (80 mg, 0.343 mmol) and (tert-butyl 2,7-diazaspiro[3.5]nonane-2-carboxylate) (94 mg, 0.412 mmol) in NMP (3 mL) was added DIPEA (133 mg, 1.03 mmol). The resulting mixture was stirred at room temperature for 2 h, the mixture was diluted with H ₂ O (50 mL) and filtered. The filter cake was washed with H ₂ O (20 mL x 2), then dried to give the title product (140 mg, 96% yield) as a light yellow solid. LCMS m/z (ESI): 424.0 [M+H] ⁺ .

Step 2: 7-(6-Chloro-7-(2-fluoro-6-hydroxyphenyl)pyrido[2,3-d]-pyrimidin-4-yl)-2,7-diazaspiro[3.5 ] Nonane-2-carboxylic acid tertiary butyl ester

To 7-(6,7-dichloropyrido[2,3-d]pyrimidin-4-yl)-2,7-diazaspiro-[3.5]nonane-2-carboxylic acid tertiary butyl ester (140 mg, 0.331 mmol) and (2-fluoro-6-hydroxyphenyl)boronic acid (156 mg, 0.496 mmol) in dioxane (5 mL) and H ₂ O (1 mL) were added K ₂ CO ₃ (92 mg, 0.662 mmol) and Pd(dppf)Cl ₂ (36 mg, 0.049 mmol). The reaction mixture was heated at 100 °C for 1 h under nitrogen atmosphere. The mixture was diluted with H ₂ O (50 mL), and extracted with EtOAc (50 mL x 3). The combined organic layers were concentrated and the residue was purified by column (DCM:MeOH=20:1) to afford the title product (100 mg, 60.6% yield) as a yellow solid. LCMS m/z (ESI): 500.1 [M+H] ⁺ .

Step 3. 2-(6-Chloro-4-(2,7-diazaspiro[3.5]non-7-yl)pyrido[2,3-d]pyrimidin-7-yl)-3-fluorophenol

To 7-(6-chloro-7-(2-fluoro-6-hydroxyphenyl)pyrido[2,3-d]-pyrimidin-4-yl)-2,7-diazaspiro[3.5]nonane To a solution of tert-butyl alkane-2-carboxylate (100 mg, 0.204 mmol) in DCM (1.0 mL) was added TFA (0.5 mL). The reaction mixture was stirred at room temperature for 1 h. The reaction was concentrated and the residue was purified by reverse phase column (H ₂ O : MeCN = 0% to 50%) to give the title product (70 mg, 88% yield) as a pale yellow solid. LCMS: MS m/z (ESI): 400.2 [M+H] ⁺ .

Step 4: (E)-1-(7-(6-chloro-7-(2-fluoro-6-hydroxyphenyl)pyrido[2,3-d]pyrimidin-4-yl)-2,7- Diazaspiro[3.5]non-2-yl)pent-2-ene-1,4-dione

At 0ºC, to 2-(6-chloro-4-(2,7-diazaspiro[3.5]non-7-yl)pyrido[2,3-d]pyrimidin-7-yl)-3- To a solution of fluorophenol (40 mg, 0.1 mmol) and (E)-4-oxopent-2-enoic acid (5.7 mg, 0.05 mmol) in DMF (1.5 mL) was added DIPEA (26 mg, 0.2 mmol 3eq) and HATU (57 mg, 0.15 mmol). The mixture was stirred at 0 ºC for 20 min. The mixture was diluted with H ₂ O (50 mL), and extracted with EtOAc (30 mL x 3). The combined organic layers were concentrated and the residue was purified by preparative HPLC to provide the title product (4.7 mg, 16.3% yield) as a white solid. LCMS: MS m/z (ESI): 496.1 [M+H] ⁺ . ¹ H NMR (400 MHz, CDCl ₃ ): δ 9.69 (s, 1H), 8.85 (s, 1H), 8.31 (s, 1H), 7.40 - 7.34 (m, 1H), 7.13 (d, J = 15.4 Hz , 1H), 6.95 (d, J = 8.4 Hz, 1H), 6.82 (d, J = 15.4 Hz, 1H), 6.78 - 6.72 (m, 1H), 4.11 (s, 2H), 3.96 (s, 2H) , 3.90 (t, J = 5.2 Hz, 2H), 3.86 - 3.78 (m, 2H), 2.36 (s, 3H), 2.08 - 1.99 (m, 4H).

Example 11

(S)-2-(4-(6-chloro-7-(8-chloronaphthalen-1-yl)pyrido[2,3-d]pyrimidin-4-yl)-1-(2-fluoroacryl Base) piperazin-2-yl) acetonitrile

Step 1: (S)-tert-butyl 2-(cyanomethyl)-4-(6,7-dichloropyrido[2,3-d]pyrimidin-4-yl)piperazine-1-carboxylate

To 4,6,7-trichloropyrido[2,3-d]pyrimidine (80 mg, 0.341 mmol) and (S)-2-(cyanomethyl)piperazine-1-carboxylic acid tertiary butyl ester ( 85 mg, 0.375 mmol) in NMP (3 mL) was added DIPEA (133 mg, 1.03 mmol). The resulting mixture was stirred at room temperature for 2 h, the mixture was diluted with H ₂ O (50 mL), and filtered. The filter cake was washed with H ₂ O (20 mL x 2), then dried to give the title product (70 mg, 48% yield) as a light yellow solid. LCMS m/z (ESI): 422.9 [M+H] ⁺ .

Step 2: (S)-4-(6-Chloro-7-(8-chloronaphthalen-1-yl)pyrido[2,3-d]pyrimidin-4-yl)-2-(cyanomethyl) tertiary butyl piperazine-1-carboxylate

To (S)-2-(cyanomethyl)-4-(6,7-dichloropyrido[2,3-d]pyrimidin-4-yl)piperazine-1-carboxylic acid tertiary butyl ester (70 mg, 0.165 mmol) and 2-(8-chloronaphthalen-1-yl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (72 mg, 0.248 mmol ) in 1,4-dioxane (5 mL) and H ₂ O (1 mL) were added K ₂ CO ₃ (46 mg, 0.330 mmol) and Pd(PPh ₃ ) ₂ Cl ₂ (12 mg, 0.017 mmol). The reaction mixture was heated at 100 °C for 1 h under nitrogen atmosphere. The mixture was diluted with H ₂ O (50 mL), and extracted with EtOAc (50 mL x 3). The combined organic layers were concentrated and the residue was purified by column (DCM:MeOH=20:1) to afford the title product (30 mg, 33% yield) as a yellow solid. LCMS m/z (ESI): 549.1 [M+H] ⁺ .

Step 3: (S)-2-(4-(6-chloro-7-(8-chloronaphthalen-1-yl)pyrido[2,3-d]pyrimidin-4-yl)piperazin-2-yl ) acetonitrile

To (S)-4-(6-chloro-7-(8-chloronaphthalen-1-yl)pyrido[2,3-d]pyrimidin-4-yl)-2-(cyanomethyl)piperazine - To a solution of tert-butyl-1-carboxylate (30 mg, 0.054 mmol) in DCM (1.0 mL) was added TFA (0.5 mL). The reaction mixture was stirred at room temperature for 1 h. The reaction was concentrated to give the title product (20 mg) as a brown gum. LCMS: MS m/z (ESI): 449.1 [M+H] ⁺

Step 4: (S)-2-(4-(6-Chloro-7-(8-chloronaphthalen-1-yl)pyrido[2,3-d]pyrimidin-4-yl)-1-(2- Fluoroacryl)piperazin-2-yl)acetonitrile

At 0ºC, to (S)-2-(4-(6-chloro-7-(8-chloronaphthalen-1-yl)pyrido[2,3-d]pyrimidin-4-yl)piperazine-2 -yl) acetonitrile (20 mg, 0.045 mmol) and 2-fluoroacrylic acid (2 mg, 0.022 eq) in DMF (2 mL) were added DIPEA (11 mg, 0.89 mmol) and HATU (17 mg, 0.045 mmol ). The mixture was stirred at 0 ºC for 20 min. The mixture was diluted with H ₂ O (5 mL), and extracted with EtOAc (10 mL). The combined organic layers were concentrated, and the residue was purified by prep-TLC (CH ₂ Cl ₂ :MeOH = 10:1) to afford the title product (0.98 mg, 4% yield) as a white solid. LCMS: MS m/z (ESI): 521.1 [M+H] ⁺ .

Example 12

1-((3S)-4-(6-chloro-7-(2-fluoro-6-hydroxyphenyl)-2-(((S)-1-methylpyrrolidin-2-yl)methoxy )pyrido[2,3-d]pyrimidin-4-yl)-3-methylpiperazin-1-yl)prop-2-en-1-one

Step 1: 4,6,7-Trichloropyrido[2,3-d]pyrimidine

To a solution of 6,7-dichloropyrido[2,3-d]pyrimidine-2,4-diol (500 mg, 2.15 mmol) in POCl ( ₁₀ mL) was added DIPEA (557 mg, 4.31 mmol ). The mixture was stirred at 100 ºC for 3 h. The mixture was concentrated and the residue was purified by column (petroleum ether:EtOAc=20:1) to afford the title product (440 mg, 76% yield) as a yellow solid. LCMS: MS m/z (ESI): 270.0 [M+H] ⁺ .

Step 2: (S)-tert-butyl 4-(6,7-dichloropyrido[2,3-d]pyrimidin-4-yl)-3-methyl-piperazine-1-carboxylate

4,6,7-trichloropyrido[2,3-d]pyrimidine (240 mg, 0.89 mmol), (S)-3-methylpiperazine-1-carboxylic acid tertiary butyl ester (197 mg, 0.98 mmol) and DIPEA (230 mg, 1.79 mmol) in NMP (5 mL) was heated at room temperature for 1 h. The mixture was diluted with H ₂ O (80 mL) and filtered. The filter cake was washed with H ₂ O (20 mL x 2), then dried to give the title product (280 mg, 72% yield) as a yellow solid. LCMS: MS m/z (ESI): 433.8 [M+H] ⁺ .

Step 3: (S)-4-(6,7-Dichloro-2-(((S)-1-methylpyrrolidin-2-yl)methoxy)pyrido[2,3-d]pyrimidine -4-yl)-3-methylpiperazine-1-carboxylic acid tertiary butyl ester

(S)-4-(6,7-dichloropyrido[2,3-d]pyrimidin-4-yl)-3-methylpiperazine-1-carboxylic acid tertiary butyl ester (100 mg, 0.232 mmol ), (S)-(1-methylpyrrolidin-2-yl)methanol (32 mg, 0.278 mmol) and DIPEA (59.86 mg, 0.464 mmol) in CH ₃ CN (4 mL) were heated at 80 ºC for 8 h. The mixture was diluted with H ₂ O (30 mL), and extracted with EtOAc (30 x 3 mL). The combined organic layers were concentrated and the residue was purified by column (CH ₂ Cl ₂ :MeOH = 20:1) to afford the title product (90 mg, 76% yield) as a yellow solid. ¹ H NMR (400 MHz, CDCl3): δ 8.03 (s, 1H), 4.64 - 4.53 (m, 1H), 4.57 - 4.33 (m, 2H), 4.05 (dd, J = 14.4, 7.2 Hz, 2H), 3.86 (s, 1H), 3.54 (s, 1H), 3.16 (s, 1H), 3.04 (t, J = 7.6 Hz, 2H), 2.62 (dd, J = 8.0, 6.2 Hz, 1H), 2.44 (s , 3H), 2.23 - 2.21 (m, 1H), 2.05 - 1.87 (m, 2H), 1.71 - 1.66 (m, 3H), 1.39 (s, 9H), 1.35 (d, J = 6.6 Hz, 3H), 1.19 (t, J = 7.2 Hz, 1H).

Step 4: (3S)-4-(6-Chloro-7-(2-fluoro-6-hydroxyphenyl)-2-(((S)-1-methylpyrrolidin-2-yl)methoxy )pyrido[2,3-d]pyrimidin-4-yl)-3-methylpiperazine-1-carboxylic acid tertiary butyl ester

(S)-4-(6,7-dichloro-2-(((S)-1-methylpyrrolidin-2-yl)methoxy)pyrido[2,3-d]pyrimidine-4 -yl)-3-methylpiperazine-1-carboxylic acid tertiary butyl ester (90 mg, 0.176 mmol), Pd(PPh ₃ ) ₂ Cl ₂ (12.3 mg, 0.017 mmol), K ₂ CO ₃ (48.2mg, 0.352 mmol) in 1,4-dioxane (5 mL) and H ₂ O (1 ml) was heated at 100 ºC for 2 h. The mixture was diluted with H ₂ O (10 mL), and extracted with EtOAc (30 mL x 3). The combined organic layers were concentrated and the residue was purified by column (DCM:MeOH=10:1) to afford the title product (80 mg, 74% yield) as a yellow solid.

Step 5: 2-(6-Chloro-4-((S)-2-methylpiperazin-1-yl)-2-(((S)-1-methylpyrrolidin-2-yl)methoxy Base) pyrido[2,3-d]pyrimidin-7-yl)-3-fluorophenol

(3S)-4-(6-chloro-7-(2-fluoro-6-hydroxyphenyl)-2-(((S)-1-methylpyrrolidin-2-yl)methoxy)pyridine And[2,3-d]pyrimidin-4-yl)-3-methylpiperazine-1-carboxylic acid tert-butyl ester (80 mg, 0.136 mmol) in 4 M HCl/1,4-dioxane (4 mL) was stirred at room temperature for 2 h. The mixture was concentrated to give the crude title product as a white solid, which was used directly in the next step without purification. LCMS: MS m/z (ESI): 487.0 [M+H] ⁺ .

Step 6: 1-((3S)-4-(6-Chloro-7-(2-fluoro-6-hydroxyphenyl)-2-(((S)-1-methylpyrrolidin-2-yl) Methoxy)pyrido[2,3-d]pyrimidin-4-yl)-3-methylpiperazin-1-yl)prop-2-en-1-one

At 0ºC, to 2-(6-chloro-4-((S)-2-methylpiperazin-1-yl)-2-(((S)-1-methylpyrrolidin-2-yl) A solution of methoxy)pyrido[2,3-d]pyrimidin-7-yl)-3-fluorophenol (80 mg, 0.16 mmol) and acrylic acid (12 mg, 0.17 mmol) in DMF (1.5 mL) DIEA (41 mg, 0.32 mmol) and HATU (91 mg, 0.24 mmol) were added. The mixture was stirred at 0°C for 30 min. The mixture was diluted with H ₂ O (50 mL), and extracted with EtOAc (150 mL). The combined organic layers were concentrated, and the residue was purified by preparative HPLC to afford the title product (32.23 mg, 37%) as a light yellow solid. LCMS: MS m/z (ESI): 541.3 [M+H] ⁺ . ¹ H NMR (400 MHz, DMSO-d ₆ ): δ 10.22 (brs, 1H), 8.44 (s, 1H), 7.33 (dd, J = 15.3, 8.0 Hz, 1H), 6.92 – 6.73 (m, 3H) , 6.21 – 6.15 (m, 1H), 5.76 – 5.73 (m, 1H), 4.82 (brs, 1H), 4.39 – 4.35 (m, 1H), 4.28 – 4.07 (m, 4H), 4.02 – 3.96 (m, 1H), 3.72 – 3.61 (m, 1H), 3.09 – 3.04 (m, 1H), 2.99 – 2.95 (m, 1H), 2.63 – 2.57 (m, 1H), 2.37 (s, 3H), 2.20 (q, J = 8.4 Hz, 1H), 1.99 – 1.92 (m, 1H), 1.73 – 1.57 (m, 3H), 1.29 (d, J = 5.8 Hz, 3H).

Example 13

(E)-1-((3S)-4-(6-chloro-7-(2-fluoro-6-hydroxyphenyl)-2-(((S)-1-methylpyrrolidin-2-yl )methoxy)pyrido[2,3-d]pyrimidin-4-yl)-3-methylpiperazin-1-yl)pent-2-ene-1,4-dione

Step 1: (E)-1-((3S)-4-(6-chloro-7-(2-fluoro-6-hydroxyphenyl)-2-(((S)-1-methylpyrrolidine- 2-yl)methoxy)pyrido[2,3-d]pyrimidin-4-yl)-3-methylpiperazin-1-yl)pent-2-ene-1,4-dione

At 0ºC, to 2-(6-chloro-4-((S)-2-methylpiperazin-1-yl)-2-(((S)-1-methylpyrrolidin-2-yl) Methoxy)pyrido[2,3-d]pyrimidin-7-yl)-3-fluorophenol (30 mg, 0.0616 mmol) and (E)-4-oxopent-2-enoic acid (7.03 mg , 0.061 mmol) in DMF (1.5 mL) were added DIEA (23.8 mg, 0.185 mmol) and HATU (35.1 mg, 0.092 mmol). The mixture was stirred at 0°C for 30 min. The mixture was diluted with H ₂ O (30 mL), and extracted with EtOAc (30 mL x 3). The combined organic layers were concentrated, and the residue was purified by preparative HPLC to afford the title product (3.3 mg, 10% yield) as a white solid. LCMS: MS m/z (ESI): 583.2 [M+H] ⁺ . ¹ H NMR (400 MHz, CD ₃ OD): δ 8.51 (s, 2H), 7.49 - 7.30 (m, 2H), 6.96 - 6-90 (m, 1H), 6.86 - 6.67 (m, 2H), 4.78 - 4.75 (m, 2H), 4.64 - 4.59 (m, 1H), 4.50 - 4.35 (m, 2H), 4.18 - 3.99 (m, 1H), 3.94 - 3.52 (m, 5H), 3.12 - 3.01 (m, 1H), 2.94 (s, 3H), 2.38 - 1.99 (m, 4H), 1.45 (s, 3H).

Example 14

1-((3S)-4-(6-chloro-7-(2-fluoro-6-hydroxyphenyl)-2-((((S)-1-methylpyrrolidin-2-yl)methyl )amino)pyrido[2,3-d]pyrimidin-4-yl)-3-methylpiperazin-1-yl)prop-2-en-1-one

Step 1: (S)-4-(6,7-Dichloro-2-((((S)-1-methylpyrrolidin-2-yl)methyl)amino)pyrido[2,3- d] pyrimidin-4-yl)-3-methylpiperazine-1-carboxylic acid tertiary butyl ester

(S)-4-(6,7-dichloropyrido[2,3-d]pyrimidin-4-yl)-3-methylpiperazine-1-carboxylic acid tertiary butyl ester (50 mg, 0.116 mmol ), (S)-(1-methylpyrrolidin-2-yl)methanamine (32 mg, 0.116 mmol) and DIEA (59.86 mg, 0.232 mmol) in CH ₃ CN (4 mL) were heated at 80 ºC 2 h. The mixture was diluted with H ₂ O (20 mL), and extracted with EtOAc (20 mL x 3). The combined organic layers were concentrated and the residue was purified by column (DCM:MeOH=20:1) to afford the title product (45 mg, 76%) as a yellow solid. LCMS: MS m/z (ESI): 510.3 M+H] ⁺ .

Step 2: (3S)-4-(6-Chloro-7-(2-fluoro-6-hydroxyphenyl)-2-((((S)-1-methylpyrrolidin-2-yl)methyl )amino)pyrido[2,3-d]pyrimidin-4-yl)-3-methylpiperazine-1-carboxylic acid tertiary butyl ester

To (S)-4-(6,7-dichloro-2-((((S)-1-methylpyrrolidin-2-yl)methyl)amino)pyrido[2,3-d] Pyrimidin-4-yl)-3-methylpiperazine-1-carboxylic acid tert-butyl ester (45 mg, 0.088 mmol), Pd(PPh ₃ ) ₂ Cl ₂ (12.3 mg, 0.0088 mmol) and K ₂ CO ₃ ( 48.2 mg, 0.176 mmol) in 1,4-dioxane (5 mL) and H ₂ O (1 ml) was heated at 100 ºC for 2 h. The mixture was diluted with H ₂ O (40 mL), and extracted with EtOAc (50 mL x 3). The combined organic layers were concentrated and the residue was purified by column (DCM/MeOH=10:1) to afford the title product (40 mg, 77% yield) as a yellow solid. LCMS: MS m/z (ESI): 586.4 M+H] ⁺ .

Step 3: 2(2-(6-Chloro-4-((S)-2-methylpiperazin-1-yl)-2-((((S)-1-methylpyrrolidin-2-yl )methyl)amino)pyrido[2,3-d]pyrimidin-7-yl)-3-fluorophenol

(3S)-4-(6-chloro-7-(2-fluoro-6-hydroxyphenyl)-2-((((S)-1-methylpyrrolidin-2-yl)methyl)amine yl)pyrido[2,3-d]pyrimidin-4-yl)-3-methylpiperazine-1-carboxylic acid tert-butyl ester (40 mg, 0.136 mmol) in 4 M HCl/1,4-diox The solution in alkanes (4 mL) was stirred at room temperature for 2 h. The mixture was concentrated to give the crude title product (30 mg) as a white solid, which was used directly in the next step without purification. LCMS: MS m/z (ESI): 486.2 [M+H] ⁺ .

Step 4: (E)-1-((3S)-4-(6-chloro-7-(2-fluoro-6-hydroxyphenyl)-2-(((S)-1-methylpyrrolidine- 2-yl)methoxy)pyrido[2,3-d]pyrimidin-4-yl)-3-methylpiperazin-1-yl)pent-2-ene-1,4-dione

At -70ºC, to 2(2-(6-chloro-4-((S)-2-methylpiperazin-1-yl)-2-((((S)-1-methylpyrrolidine- 2-yl)methyl)amino)pyrido[2,3-d]pyrimidin-7-yl)-3-fluorophenol (30 mg, 0.061 mmol) and DIEA (23.8 mg, 0.185 mmol) in THF (1.5 mL) was added acryloyl chloride (7.03 mg, 0.061 mmol). The mixture was stirred at -70 ºC for 10 min. The mixture was diluted with H ₂ O (20 mL) and extracted with EtOAc (20 mL x 3). The combined organic layers were concentrated and the residue was purified by preparative HPLC to afford the title product (3.3 mg, 11%) as a white solid. LCMS: MS m/z (ESI): 540.4 [M+H] ⁺ . ¹ H NMR (400 MHz, CD ₃ OD): δ 8.07 (s, 1H), 7.28 (td, J = 8.2, 6.0 Hz, 1H), 6.88 - 6.70 (m, 2H), 6.62 (dd, J = 11.8 , 8.0 Hz, 1H), 6.27 (dd, J = 16.0, 9.6 Hz, 1H), 5.79 (d, J = 10.4 Hz, 1H), 4.50 -3.90 (m, 4H), 3.72 - 3.47 (m, 4H) , 3.22 - 3.10 (m, 2H), 2.77 (s, 1H), 2.56 (s, 3H), 2.38 - 2.36 (m, 1H), 2.12 - 1.96 (m, 1H), 1.89 - 1.71 (m, 3H) , 1.41 (d, J = 6.7 Hz, 3H).

Example 15

1-(5-(6-chloro-7-(2-fluoro-6-hydroxyphenyl)-2-(((S)-1-methylpyrrolidin-2-yl)methoxy)pyrido[ 2,3-d]pyrimidin-4-yl)-2,5-diazabicyclo[4.1.0]hept-2-yl)prop-2-en-1-one

Step 1: 5-(2,6,7-Trichloropyrido[2,3-d]pyrimidin-4-yl)-2,5-diazabicyclo[4.1.0]heptan-2-carboxylic acid tertiary Butyl ester

To 2,4,6,7-tetrachloropyrido[2,3-d]pyrimidine (128 mg, 0.476 mmol) and (2,5-diazabicyclo[4.1.0]heptane-2-carboxylic acid tris To a solution of butyl ester) (40 mg, 0.571 mmol) in NMP (3 mL) was added DIEA (80 mg, 1.427 mmol). The resulting reaction was stirred at room temperature for 2 h. The mixture was diluted with H ₂ O (80 mL), and filtered. The filter cake was washed with H ₂ O (20 mL x 2), then dried to give the title product (90 mg, 44% yield) as a yellow solid. LCMS: m/z (ESI) 429.9 [M+H] ⁺ .

Step 2: 5-(6,7-Dichloro-2-(((S)-1-methylpyrrolidin-2-yl)methoxy)pyrido[2,3-d]pyrimidin-4-yl )-2,5-diazabicyclo[4.1.0]heptane-2-carboxylic acid tertiary butyl ester

5-(2,6,7-trichloropyrido[2,3-d]pyrimidin-4-yl)-2,5-diazabicyclo[4.1.0]heptane-2-carboxylic acid tertiary butane To a solution of ester (90 mg, 0.212 mmol), (S)-(1-methylpyrrolidin-2-yl)methanol (27 mg, 0.232 mmol, 1.2 eq) in CHCN ( ₅ mL) was added DIPEA (55 mg, 0.423 mmol, 2.0 eq). The reaction mixture was heated at 80 ºC for 12 h under nitrogen atmosphere. The reaction mixture was concentrated and the residue was purified by column (CH ₂ Cl ₂ :MeOH = 30:1) to give the title product (80 mg, 75.5% yield) as a yellow solid. LCMS m/z (ESI): 509.2 [M+H] ⁺ .

Step 3: 5-(6-chloro-7-(2-fluoro-6-hydroxyphenyl)-2-(((S)-1-methylpyrrolidin-2-yl)methoxy)pyrido[ 2,3-d]pyrimidin-4-yl)-2,5-diazabicyclo[4.1.0]heptane-2-carboxylic acid tertiary butyl ester

To 5-(6,7-dichloro-2-(((S)-1-methylpyrrolidin-2-yl)methoxy)pyrido[2,3-d]pyrimidin-4-yl)- 2,5-diazabicyclo[4.1.0]heptane-2-carboxylic acid tert-butyl ester (80 mg, 0.157 mmol), (2-fluoro-6-hydroxyphenyl)boronic acid (37 mg, 0.232 mmol, 1.5 eq) To a solution in dioxane (5 mL) and H ₂ O (1 mL) was added K ₂ CO ₃ (44 mg, 0.315 mmol) and Pd(dppf)Cl ₂ (18 mg, 0.015 mmol). The reaction mixture was heated at 100 °C for 2 h under nitrogen atmosphere. The reaction was concentrated and the residue was purified by column (CH ₂ Cl ₂ :MeOH = 10:1) to give the title product (20 mg, 22.3% yield) as a yellow solid. LCMS m/z (ESI): 585.2 [M+H] ⁺ .

Step 4: 2-(4-(2,5-Diazabicyclo[4.1.0]hept-2-yl)-6-chloro-2-(((S)-1-methylpyrrolidin-2- Base)methoxy)pyrido[2,3-d]pyrimidin-7-yl)-3-fluorophenol

To 5-(6-chloro-7-(2-fluoro-6-hydroxyphenyl)-2-(((S)-1-methylpyrrolidin-2-yl)methoxy)pyrido[2, 3-d]pyrimidin-4-yl)-2,5-diazabicyclo-[4.1.0]heptane-2-carboxylic acid tert-butyl ester (20 mg, 0.034 mmol) in DCM (0.5 mL) TFA (0.2 mL) was added to the solution. The reaction mixture was stirred at room temperature for 1 h. The reaction was concentrated to give the title product (15 mg) as a brown gum which was used in the next step without purification. LCMS: MS m/z (ESI): 485.1 [M+H] ⁺ .

Step 5: 1-(5-(6-Chloro-7-(2-fluoro-6-hydroxyphenyl)-2-(((S)-1-methylpyrrolidin-2-yl)methoxy) Pyrido[2,3-d]pyrimidin-4-yl)-2,5-diazabicyclo[4.1.0]hept-2-yl)prop-2-en-1-one

At -70ºC, the compound 2-(4-(2,5-diazabicyclo[4.1.0]hept-2-yl)-6-chloro-2-(((S)-1-methylpyrrole Pyridin-2-yl)methoxy)pyrido[2,3-d]pyrimidin-7-yl)-3-fluorophenol (15 mg, 0.03 mmol) and DIPEA (2 mg, 0.09 mmol) in DCM (2.0 mL) was added acryloyl chloride (1.5 mg, 0.015 mmol). The mixture was stirred at -70 ºC for 10 min. The reaction mixture was diluted with H ₂ O (10 mL), and extracted with EtOAc (10 mL x 3). The combined organic layers were concentrated and the residue was purified by preparative HPLC to afford the title product (1.03 mg, 4.5% yield) as a white solid. LCMS: MS m/z (ESI): 539.3 [M+H] ⁺ . ¹ H NMR (400 MHz, CDCl ₃ ): δ 8.88 (s, 1H), 7.40 - 7.30 (m, 2H), 6.90 - 6.90 (m, 1H), 6.88 - 6.66 (m, 2H), 6.45 (d, J = 16.0 Hz, 1H), 5.78 (d, J = 16.0 Hz, 1H), 4.88 - 4.58 (m, 2H), 4.04 - 3.85 (m, 1H), 3.83 - 3.25 (m, 7H), 2.92 (s , 3H), 2.33 - 1.96 (m, 5H), 0.88 - 0.79 (m, 2H).

Example 16

(E)-1-(7-(6-chloro-7-(2-fluoro-6-hydroxyphenyl)-2-(((S)-1-methylpyrrolidin-2-yl)methoxy )pyrido[2,3-d]pyrimidin-4-yl)-2,7-diazaspiro[3.5]non-2-yl)pent-2-ene-1,4-dione

Step 1: 7-(2,6,7-Trichloropyrido[2,3-d]pyrimidin-4-yl)-2,7-diazaspiro[3.5]nonane-2-carboxylic acid tert-butyl ester

To 2,4,6,7-tetrachloropyrido[2,3-d]pyrimidine (50 mg, 0.187 mmol) and 2,7-diazaspiro[3.5]nonane-2-carboxylic acid tertiary butyl To a solution of the ester (42.3 mg, 0.187) in NMP (3 ml) was added DIEA (48.3 mg, 0.374 mmol). The reaction was stirred at room temperature for 1 h. The mixture was diluted with H ₂ O (20 mL), and extracted with EtOAc (30 x 3 mL). The combined organic layers were concentrated and the residue was purified by column (DCM:MeOH=20:1) to afford the title product (60 mg, 70%) as a yellow solid.

LCMS: MS m/z (ESI): 458.1 [M+H] ⁺ .

Step 2: (S)-7-(6,7-Dichloro-2-((1-methylpyrrolidin-2-yl)methoxy)pyrido[2,3-d]pyrimidin-4-yl )-2,7-diazaspiro[3.5]nonane-2-carboxylate

7-(2,6,7-trichloropyrido[2,3-d]pyrimidin-4-yl)-2,7-diazaspiro[3.5]nonane-2-carboxylic acid tertiary butyl ester ( 60 mg, 0.130 mmol), (S)-(1-methylpyrrolidin-2-yl)methanol (18.7 mg, 0.157 mmol), DIEA (33.9 mg, 0.262 mmol) in CH ₃ CN (4 mL) The solution was heated at 80ºC for 8 h. The mixture was diluted with H ₂ O (50 mL), and extracted with EtOAc (50 x 3 mL). The combined organic layers were concentrated, and the residue was purified by column (DCM:MeOH=20:1) to afford the title product (50 mg, 71% yield) as a yellow solid. LCMS: MS m/z (ESI): 537.2 [M+H] ⁺ .

Step 3: 7-(6-Chloro-7-(2-fluoro-6-hydroxyphenyl)-2-(((S)-1-methylpyrrolidin-2-yl)methoxy)pyrido[ 2,3-d]pyrimidin-4-yl)-2,7-diazaspiro[3.5]nonane-2-carboxylic acid tertiary butyl ester

(S)-7-(6,7-dichloro-2-((1-methylpyrrolidin-2-yl)methoxy)pyrido[2,3-d]pyrimidin-4-yl)- 2,7-diazaspiro[3.5]nonane-2-carboxylate (50 mg, 0.093 mmol), Pd(PPh ₃ ) ₂ Cl ₂ (6.7 mg, 0.0093 mmol), K ₂ CO ₃ (25.6 mg , 0.186 mmol) in 1,4-dioxane (5 mL) and H ₂ O (1 ml) was heated at 100 ºC for 1 h. The mixture was diluted with H ₂ O (30 mL), and extracted with EtOAc (30 x 3 mL). The combined organic layers were concentrated and the residue was purified by column (DCM/MeOH=10:1) to provide the title product (40 mg, 74% yield). LCMS: MS m/z (ESI): 613.2 [M+H] ⁺ .

Step 4: 2-(6-Chloro-2-(((S)-1-methylpyrrolidin-2-yl)methoxy)-4-(2,7-diazaspiro[3.5]nonane- 7-yl)pyrido[2,3-d]pyrimidin-7-yl)-3-fluorophenol

7-(6-chloro-7-(2-fluoro-6-hydroxyphenyl)-2-(((S)-1-methylpyrrolidin-2-yl)methoxy)pyrido[2, 3-d]pyrimidin-4-yl)-2,7-diazaspiro[3.5]nonane-2-carboxylic acid tert-butyl ester (40 mg, 0.065 mmol) in 4 M HCl/1,4-diox The solution in alkanes (2 mL) was stirred at room temperature for 1 h. The mixture was concentrated to give the crude title product (20 mg) as a white solid, which was used directly in the next step without purification. LCMS: MS m/z (ESI): 513.0 [M+H] ⁺ .

Step 5: (E)-1-(7-(6-Chloro-7-(2-fluoro-6-hydroxyphenyl)-2-(((S)-1-methylpyrrolidin-2-yl) Methoxy)pyrido[2,3-d]pyrimidin-4-yl)-2,7-diazaspiro[3.5]non-2-yl)pent-2-ene-1,4-dione

At 0ºC, 2-(6-chloro-2-(((S)-1-methylpyrrolidin-2-yl)methoxy)-4-(2,7-diazaspiro[3.5] Non-7-yl)pyrido[2,3-d]pyrimidin-7-yl)-3-fluorophenol (20 mg, 0.039 mmol) and (E)-4-oxopent-2-enoic acid ( 4.4 mg, 0.014 mmol) in DMF (1.5 mL) were added DIEA (15.1 mg, 0.117 mmol) and HATU (22.2 mg, 0.058 mmol). The mixture was stirred at 0 ºC for 20 min. The mixture was diluted with H ₂ O (20 mL), and extracted with EtOAc (20 mL x 3). The combined organic layers were concentrated, and the residue was purified by preparative HPLC to afford the title product (1.3 mg, 5% yield) as a yellow solid. LCMS: MS m/z (ESI): 609.4 [M+H] ⁺ . ¹ H NMR (400 MHz, CD ₃ OD) δ 8.47 (s, 1H), 7.31 (s, 1H), 6.94 (d, J = 6.0 Hz, 1H), 6.85 - 6.63 (m, 3H), 4.52 - 4.43 (m, 2H), 4.26 - 4.18 (m, 2H), 4.03 - 3.82 (m, 9H), 2.52 (s, 3H), 2.37 (s, 3H), 2.08 - 1.99 (m, 3H), 1.88 - 1.83 (m, 3H), 1.36 - 1.23 (m, 4H).

Example 17

1-((S)-4-(7-(6-Amino-4-methyl-3-(trifluoromethyl)pyridin-2-yl)-6-chloro-2-(((S)- 1-methylpyrrolidin-2-yl)methoxy)pyrido[2,3-d]pyrimidin-4-yl)-3-methylpiperazin-1-yl)prop-2-en-1- ketone

Example 18

(3S)-4-(6-chloro-7-(2-fluoro-6-hydroxyphenyl)-2-(((S)-1-methylpyrrolidin-2-yl)methoxy)pyrido [2,3-d]pyrimidin-4-yl)-3-methylpiperazine-1-carbonitrile

Step 1: (3S)-4-(6-Chloro-7-(2-fluoro-6-hydroxyphenyl)-2-(((S)-1-methylpyrrolidin-2-yl)methoxy )pyrido[2,3-d]pyrimidin-4-yl)-3-methylpiperazine-1-carbonitrile

At 0ºC, to 2-(6-chloro-4-((S)-2-methylpiperazin-1-yl)-2-(((S)-1-methylpyrrolidin-2-yl) Methoxy)pyrido[2,3-d]pyrimidin-7-yl)-3-fluorophenol (30 mg, 0.061 mmol) and K2CO3 ( ₁₇ mg, 0.122 mmol) in DMF ( ₃ mL) To a solution of BrCN (6.5 mg, 0.061 mmol) was added, and the mixture was stirred at 0 ºC under N ₂ for 1 h. The mixture was diluted with H ₂ O (30 mL), and extracted with EtOAc (30 mL x 3). The combined organic layers were concentrated, and the residue was purified by preparative HPLC to afford the title product (1.7 mg, 5% yield) as a white solid. LCMS: MS m/z (ESI): 512.3 [M+H] ⁺ . ¹ H NMR (400 MHz, CD ₃ OD): δ 8.48 (m, 2H), 7.32 (dd, J = 15.2, 7.6 Hz, 1H), 6.80-6.60 (m, 2H), 4.75-4.51 (m, 3H ), 4.26 (d, J = 13.2 Hz, 1H), 3.88 (t, J = 11.2 Hz, 1H), 3.61-3.30 (m, 6H), 3.02-3.00 (m, 1H), 2.91 (s, 3H) , 2.33-2.30 (m, 1H), 2.17-1.88 (m, 4H), 1.60 (d, J = 6.6 Hz, 3H).

Example 19

(S, E)-1-(4-(7-(6-amino-4-methyl-3-(trifluoromethyl)pyridin-2-yl)-6-chloropyrido[2,3- d] pyrimidin-4-yl)-3-methylpiperazin-1-yl)pent-2-ene-1,4-dione

Example 20

(E)-1-((S)-4-(7-(6-amino-4-methyl-3-(trifluoromethyl)pyridin-2-yl)-6-chloro-2-(( (S)-1-methylpyrrolidin-2-yl)methoxy)pyrido[2,3-d]pyrimidin-4-yl)-3-methylpiperazin-1-yl)pent-2- ene-1,4-dione

Example 21

(E)-1-(5-(6-chloro-7-(2-fluoro-6-hydroxyphenyl)-2-(((S)-1-methylpyrrolidin-2-yl)methoxy )pyrido[2,3-d]pyrimidin-4-yl)-2,5-diazabicyclo[4.1.0]hept-2-yl)pent-2-ene-1,4-dione

Example 22

(E)-1-((3S)-4-(6-fluoro-7-(2-fluoro-6-hydroxyphenyl)-2-(((S)-1-methylpyrrolidin-2-yl )methoxy)pyrido[2,3-d]pyrimidin-4-yl)-3-methylpiperazin-1-yl)pent-2-ene-1,4-dione

Example 23

1-((3S)-4-(6-fluoro-7-(2-fluoro-6-hydroxyphenyl)-2-(((S)-1-methylpyrrolidin-2-yl)methoxy )pyrido[2,3-d]pyrimidin-4-yl)-3-methylpiperazin-1-yl)prop-2-en-1-one

Example 24

(E)-1-((3S)-4-(7-(2-amino-3,5-dichloro-6-fluorophenyl)-6-chloro-2-(((S)-1- Methylpyrrolidin-2-yl)methoxy)pyrido[2,3-d]pyrimidin-4-yl)-3-methylpiperazin-1-yl)pent-2-ene-1,4- diketone

Example 25

1-((3S)-4-(7-(2-amino-3,5-dichloro-6-fluorophenyl)-6-chloro-2-(((S)-1-methylpyrrolidine -2-yl)methoxy)pyrido[2,3-d]pyrimidin-4-yl)-3-methylpiperazin-1-yl)prop-2-en-1-one

pharmacological test

1. KRAS 4B-(G12C) modified LC-MS determination

KRAS 4B-(G12C) modification LC-MS assay was performed by Pharmaron.

1) Assay buffer: Low Mg2+ buffer 2* GDP loading buffer 10* incubation buffer 20 mM Hepes, pH 7.5 20 mM Hepes, pH 7.5 125 mM Hepes, pH 7.5 50 mM NaCl 50 mM NaCl 750 mM NaCl 0.5 mM MgCl2 0.5 mM MgCl2 10 mM MgCl ₂ 10 mM EDTA 2 mM DTT GDP: 20 times excess to KRAS

2) Procedure:

2.1) Load GDP into KRAS-4B-G12C protein:

a) Purify KRAS-4B-G12C in Pharmaron (batch number 20200304, stock solution 1.76 mg/ml, MW=20198, concentration 87 uM) in low Mg2+ buffer

b) Dilute the protein 2-fold to 43.5 uM

c) Add 1 mL of 2*GDP loading buffer to 1 mL of 43.5 uM KRAS-4B-G12C and mix gently.

d) Incubate 2 mL of the mixture at room temperature for 1.5 h.

e) Aliquot into 100 uL/tube, snap freeze in liquid nitrogen and store at -80ºC.

2.2) KRAS-4B-G12C modification assay:

a) Prepare the reaction mixture according to the following table:

b) Dilute KRAS-4B-G12C loaded with GDP to 20 uM with 1*incubation buffer and prepare the reaction mixture as shown in the table below: volume KRAS-4B-G12C (20 Um) loaded with GDP 5uL Compound (10% DMSO solution) 5uL 10*incubation buffer 5uL MilliQ H2O 35uL total 50uL

c) Incubate at room temperature for 5 min and 30 min, respectively.

d) Add 5 ul of 5% formic acid to quench the reaction.

2.3) LC-MS test:

A total of 55 uL of the reaction mixture was centrifuged at 15,000 rpm for 10 min before injection for LC-MS testing.

a) UPLC conditions: project condition instrument: Waters Acquity I Class column: Sepax Bio-C4, 2.1 x 50 mm, 3 μm Flow rate: 0.6 mL/min Load volume: 10uL Mobile phase: A: 0.1% formic acid in water B: 0.1% formic acid in ACN temperature: 60ºC Detection: TOF MS

b) Gradient schedule for LC: time (min) A(%) B(%) 0 95 5 0.75 95 5 1 75 25 6 50 50 6.25 0 100 7.5 0 100 7.75 95 5 9 95 5

c) TOF MS parameters: project condition instrument Xevo G2-XS Qtof Capillary voltage (kV) 2.5 Sampling cone voltage (V) 40 Source temperature (ºC) 120 Cone gas flow(L/h) 50 Desolvation gas flow(L/h) 600 interface type ES, Positive analyzer mode sensitivity scan range 500-2000m/z

3). Calculation of binding % of KRAS(G12C):

% bound to KRAS(G12C) = peak height of complex/[peak height of complex + peak height of unbound KRAS G12C] X 100.

The results of the KRAS G12C binding assay are in the table below.

Table 1: Results of KRAS G12C binding assay instance number % bound to KRAS(G12C) instance number % bound to KRAS(G12C) Example 1 + Example 11 + Example 2 + Example 12 +++ Example 3 + Example 13 +++ Example 4 + Example 14 + Example 5 + Example 15 ++ Example 6 + Example 16 + Example 7 + Example 17 + Example 8 +++ Example 18 + Example 10 + +++ means >85%, ++ means <85% and >40%, and + means <40%

none

none.

Claims (14)

A compound of formula I, a pharmaceutically acceptable salt thereof, or a stereoisomer thereof,

(I) wherein R ₁ , R ₂ , R ₃ and R ₄ are each independently selected from hydrogen, alkyl and halogen, wherein R ₅ is selected from -NH ₂ and -OH, and R ₄ and R ₅ can be attached to them The connected groups together form a five-membered or six-membered ring, wherein X and Y are each independently selected from N and C, wherein L is selected from H, N, O, _SO and S, wherein Q is selected from alkyl and optionally Substituted nitrogen-containing five-membered rings, with the proviso that when L is H, Q is absent, wherein

selected from 6-membered heteroaryl groups containing 1 or 2 heteroatoms selected from N, or nitrogen-containing bicyclic rings, each of which is optionally replaced by hydrogen, -C _1-3 alkyl, halogen, -CN, 3,4, 5,6-membered carbocyclic ring substitution, wherein W is selected from -CN, optionally substituted 2-propene-1-one, or optionally substituted pent-2-ene-1,4-dione, the Optional substituents are selected from halogen, alkyl, -CN, 3-membered carbocycles. The compound of formula I as described in Claim 1, its pharmaceutically acceptable salt or its stereoisomer, wherein R ₁ , R ₂ , R ₃ and R ₄ are each independently selected from hydrogen, alkyl and halogen, wherein R is selected from -NH and _-OH , and R and R may form a _five- or _{six -} membered ring with the group to which they are attached, wherein X and Y are each independently selected from N and C, wherein L is selected from H, N, O, SO ₂ and S, wherein Q is selected from methyl and

, provided that when L is H, Q does not exist, where

selected from

,

,

,

,

,

,

with

, where W is selected from

,

,

,

,

,

,

,

with

. The compound of formula I as described in claim 1 or 2, its pharmaceutically acceptable salt or its stereoisomer, wherein R ₁ and R ₂ are each independently selected from halogen, R ₃ and R ₄ are each independently selected from From hydrogen and alkyl, wherein R is _-OH , wherein X and Y are each C, wherein L is selected from H, O and S, wherein Q is selected from

, in

selected from

,

,

,

,

,

,

with

, where W is selected from

,

,

,

,

,

,

,

with

. The compound of formula I as described in any one of claims 1-3, its pharmaceutically acceptable salt or stereoisomer thereof, wherein R ₁ and R ₂ are each independently selected from F and Cl, R ₃ and R ₄ are each independently selected from hydrogen and C _1-3 alkyl, wherein R ₅ is -OH, wherein X and Y are each C, wherein L is selected from H, O and S, wherein Q is selected from

, in

selected from

,

,

,

with

, where W is selected from

,

,

,

,

,

with

. A compound of formula I as described in any one of claims 1-4, a pharmaceutically acceptable salt thereof or a stereoisomer thereof, wherein L is selected from O and S, and Q is selected from

. A compound of formula I as described in any one of claims 1-5, a pharmaceutically acceptable salt thereof or a stereoisomer thereof, wherein L is selected from O and S, and Q is selected from

,

selected from

. A compound of formula I as described in any one of claims 1-4, a pharmaceutically acceptable salt thereof or a stereoisomer thereof, wherein L is H,

selected from

. The compound of formula I as described in any one of claims 1-7, its pharmaceutically acceptable salt or its stereoisomer, wherein W is selected from

with

. The compound of formula I as described in any one of claims 1-8, its pharmaceutically acceptable salt or its stereoisomer, wherein when L is H and

selected from

, W is

. A compound of formula I as described in any one of claims 1-9, a pharmaceutically acceptable salt thereof or a stereoisomer thereof, wherein when L is selected from O and S, Q is selected from

,and

selected from

, W selected from

with

. A compound, a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, wherein the compound is selected from the group consisting of

. A pharmaceutical composition comprising a compound of formula I as described in any one of claim items 1-11, a pharmaceutically acceptable salt thereof or a stereoisomer thereof and one or more pharmaceutically acceptable carriers, diluent or excipient. A method of treating various disorders, comprising administering the compound of formula I, its pharmaceutically acceptable salt or its stereoisomer as described in any one of claims 1-11 to a subject in need. The method of claim 13, wherein the disorder is cancer.