WO2023213269A1

WO2023213269A1 - Amide-substituted heterocyclic compounds as kras g12d modulators and uses thereof

Info

Publication number: WO2023213269A1
Application number: PCT/CN2023/092022
Authority: WO
Inventors: Lanqi Jia; Yong He; Wenting Wu; Shangfeng LI; Yugui Gu
Original assignee: Zai Lab Shanghai Co Ltd
Current assignee: Zai Lab Shanghai Co Ltd
Priority date: 2022-05-06
Filing date: 2023-05-04
Publication date: 2023-11-09
Anticipated expiration: 2024-11-06

Abstract

Disclosed are compounds of formula (I), which can be inhibitors of KRAS G12D, and their uses.

Description

AMIDE-SUBSTITUTED HETEROCYCLIC COMPOUNDS AS KRAS G12D MODULATORS AND USES THEREOF

FIELD

This application relates to KRAS G12D modulators, their preparation and uses thereof.

BACKGROUND

Rat sarcoma (RAS) , encoded by the proto-oncogenes HRAS, NRAS and KRAS, is a GTP-binding protein that is in an activated state when binding with GTP, and is in an inactive state when binding with GDP. RAS is distributed on the inner surface of the cell membrane and is activated when it binds to GTP and inactivated when it binds to GDP. The upstream of RAS is receptor tyrosine kinase (RTK) , which regulates downstream signaling pathways such as PI3K and RAF after activation, thereby regulating cell growth, survival, migration and differentiation functions. Since RAS proteins are central to the axis of many important cellular signaling networks, and these signals are associated with multiple tumor markers, overactive RAS signaling may ultimately lead to tumorigenesis.

Among RAS family members, oncogenic mutations are most commonly found in KRAS (85%) , and aberrant expression of KRAS accounts for up to 20%of all cancers, with G12D mutations accounting for 25%of pancreatic cancer (PDAC) , colon cancer (CRC) 13.3%, rectal cancer (RC) 10.1%, non-small cell lung cancer (NSCLC) 4.1%.

Although there is a high clinical need, so far there is no drug that directly targets the KRAS G12D mutation on the market. There are two main difficulties in the development of KRAS G12D inhibitors. On the one hand, the RAS protein has a smooth structure and there is no obvious pocket on the surface for small molecules to bind to; on the other hand, the affinity of KRAS protein for GTP is as high as picomolar level andendogenous GTP levels are high. Thus, it is difficult for small-molecule drugs to block the binding of the two. At present, no targeted drug for KRAS G12D mutation has entered the clinical research stage, and there is a large unmet clinical need.

SUMMARY

Provided is a compound of formula (I) or a pharmaceutically acceptable salt thereof:

wherein

Q¹, Q², and Q³ are independently selected from N, CH, C-CF₃, C-OH, C-Cl, C-F, C-CH₃, C-CH (CH₃) ₂, C-OCH₃, C-SCF₃, C-OCF₃, and C-CN;

Z is

ring A is a 4-10 membered heterocyclyl or a8-10 membered fused heteroaryl, provided when

ring A is a 8-10 membered fused heteroaryl, R⁶-L₁ is optionally present; n is 0, 1, or 2;

ring B is a 4, 5, 6, or 7 membered heterocycle and together with ring A forms a spiral bicyclic

wherein ring B contains a moiety selected from ring B is further optionally substituted with one or more W;

R¹ is selected from L-5-12 membered heterocyclyl and L-C_3-8 cycloalkyl, wherein each of the 5-12 membered heterocyclyland C_3-8 cycloalkylis optionally substituted with one or more W;

L and L₁ are independently selected from -CH₂-, -CH (CH₃) -, CH₂-CH₂-, and a bond;

Y is selected from a bond and -O-;

R² is H or C_1-3 alkyl, R⁵ is C_1-6alkyl substituted with R⁶;

R⁶ is selected from

R³ is selected from aryl and heteroaryl, wherein the aryl and heteroaryl is optionally substituted with one or more W;

R⁴ is selected from H, -CF₃, -OH, OMe, OEt, -CH₃, F, and Cl;

R⁷ is selected from H, C_1-6 alkyl optionally substituted with W, aryl optionally substituted with one or more W, cycloalkyl optionally substituted with one or more W, and 5, 6, or 7 membered heterocyclyl optionally substituted with one or more W, provided that the point of attachment of the heterocyclyl to the carbonyl carbon or sulfone sulfur is a carbon atom;

R⁸ is selected from H and C_1-6alkyl;

R⁹ is selected from H and C_1-6 alkyl; or R⁷ and R⁹, together with the nitrogen to which they are attached, form a 5, 6, or 7 membered heterocyclyl optionally substituted with one or more W; and

W is selected from OH, CN, halo, C_2-4 alkynyl, NH₂, mono or di-C_1-6 alkyl substituted amino, C_2- ₄ alkenyl, C_3-8 cycloalkyl optionally substituted with 1-5 F or Cl, C_1-6 alkoxy optionally substituted with 1-5 F or Cl, and C_1-6 alkyl optionally substituted with one or more groups selected from F, Cl, -OC (O) NR^7’ R^9’ , and -NR^7’ R^9’ , with R^7’a nd R^9’ independently being selected from H and C_1-6 alkyl, or R^7’a nd R^9’ , together with the nitrogen to which they are attached, form a 5, 6, or 7 membered heterocyclyl optionally substituted with one or more groups selected from F, Cl, C_1-6 alkyl.

In some embodiments, Q¹ is N, one of Q² and Q³ is N, the other is not. In some embodiments, Q¹ is N, Q² is N, and Q³ is not N. In some embodiments, Q¹ is N, Q² is N, and Q³ is C-H or C-F. In some embodiments, Q¹ is N, Q² is selected from N, CH, C-CF₃, C-OH, C-Cl, C-F, C-CH₃, C-CH (CH₃) ₂, C-OCH₃, C-SCF₃, C-OCF₃, and C-CN, and Q³ is not N. In some embodiments, Q¹ is N, Q² is selected from N, CH, C-CF₃, C-OH, C-Cl, C-F, C-CH₃, C-CH (CH₃) ₂, C-OCH₃, C-SCF₃, C-OCF₃, and C-CN, and Q³ is C-H or C-F. In some embodiments, Q¹ is N, Q² is selected from CH, C-CF₃, C-OH, C-Cl, C-F, C-CH₃, C-CH (CH₃) ₂, C-OCH₃, C-SCF₃, C-OCF₃, and C-CN, and Q³ is C-H or C-F.

In some embodiments, R³ is phenyl or naphthyl optionally substituted with one or more W. In some embodiments, R³ is pyridinyl, 1H-indazolyl, or 1H-indolyl, each of which is optionally substituted with one or more W. In some embodiments, R³ is phenyl, naphthyl, pyridinyl, benzothiazolyl, benzothiophenyl, 1H-indenyl, 1H-indazolyl, or 1H-indolyl, each of which is optionally substituted with one or more W. In some embodiments, R³ is phenyl, naphthyl, pyridinyl, benzothiazolyl, benzothiophenyl, 1H-indenyl, 1H-indazolyl, or 1H-indolyl, each of which is optionally substituted with one or more groups selected from F, Cl, methyl, ethyl, ethenyl, ethynyl, CF₃, cyclopropyl, iso-propyl, -NH₂, -CN, and OH.

In some embodiments, R³ is selected from

In some embodiments, Y is O.

In some embodiments, R¹ is selected from -L-fused 7-10 membered bicyclic heterocyclyl comprising at least one nitrogen, and -L-5-7 membered monocyclic heterocyclyl comprising at least one nitrogen, each of the fused 7-10 membered bicyclic heterocyclyl comprising at least one nitrogen and 5-7 membered monocyclic heterocyclyl comprising at least one nitrogen is optionally substituted with one or more W.

In some embodiments, each of the fused 7-10 membered bicyclic heterocyclyl comprising at least one nitrogen and 5-7 membered monocyclic heterocyclyl comprising at least one nitrogen is optionally substituted with one or more group selected from F and C_1-3 alkyloptionally substituted with -OC (O) NR^7’ R^9’ or -NR^7’ R^9’ , with R^7’a nd R^9’ independently being selected from H and C_1-6 alkyl, or R^7’a nd R^9’ , together with the nitrogen to which they are attached, form a 5, 6, or 7 membered heterocyclyl optionally substituted with one or more groups selected from F, Cl, C_1-6 alkyl. In some embodiments, R^7’a nd R^9’a re methyl or R^7’a nd R^9’ , together with the nitrogen to which they are attached, form a morpholine.

In some embodiments, R¹ is L-C_3-8 cycloalkyl optionally substituted with one or more W. In some embodiments, R¹ isL-C_3-8 cycloalkyl optionally substituted with methyl, wherein the methyl is optionally substituted with -OC (O) NR^7’ R^9’ or -NR^7’ R^9’ , with R^7’a nd R^9’ independently being selected from H and C_1-6 alkyl, or R^7’a nd R^9’ , together with the nitrogen to which they are attached, form a 5, 6, or 7 membered heterocyclyl optionally substituted with one or more groups selected from F, Cl, C_1-6 alkyl. In some embodiments, R¹is L-C_3-8 cycloalkyl optionally substituted with methyl, wherein the methyl is optionally substituted with -O (CO) N (CH₃) ₂, -O (CO) -morpholine, -N (CH₃) ₂, or morpholine.

In some embodiments, R¹ is

In some embodiments, L is CH₂or CH (CH₃) .

In some embodiments, Z iswherein R² is H or C_1-3 alkyl, R⁵ is C_3-6 alkyl substituted with R⁶.

In some embodiments, Z iswherein R² is H, R⁵ is propyl substituted with R⁶.

In some embodiments, Z is

In some embodiments, Z iswherein ring A is a 4-10 membered heterocyclylhaving only one atom on the ring is a heteroatom selected from N, O, and S or ring A is a 8-10 membered fused bi-cyclic heteroaryl with R⁶-L₁ being optionally present

In some embodiments, ring A is a 4-10 membered heterocyclyl wherein the only one heteroatom on the ring is N.

In some embodiments, ring A is a 4-10 membered heterocyclyl wherein the only one heteroatom on the ring is N and the N is the point of attachment as indicated by

In some embodiments, ring A is

In some embodiments, ring A is a 8-10 membered fused bi-cyclic heteroaryl wherein one ring is aromatic and the other is not aromatic. In some embodiments, ring A is a 8-10 membered fused bi-cyclic heteroaryl wherein one ring is aromatic, the other is not aromatic and is the point of attachment as indicated by

In some embodiments, ring A is

In some embodiments, L₁ is a bond or CH₂.

In some embodiments, n is 0. In some embodiments, n is 1 and W is selected from OH, methyl, and ethyl.

In some embodiments, Z iswherein ring A is a 4-10 membered heterocyclyl having only one atom on the ring is a heteroatom selected from N, O, and S.

In some embodiments, ring A is

In some embodiments, n is 0. In some embodiments, n is 1 and W is methyl or OH.

In some embodiments, ring B is a 5 or 6 membered heterocycle containing a moiety selected from

In some embodiments, ring B is a 5 or 6 membered heterocycle containing a moiety selected from wherein R⁸ and R⁹ are independently selected from H, methyl, and ethyl. In some embodiments, ring B is a 5 membered heterocycle containing wherein R⁸ and R⁹ are independently selected from H, methyl, and ethyl. In some embodiments, ring B is a 4 membered heterocycle containing

In some embodiments, ring B is a 5 or 6 membered heterocycle containingwherein R⁸is selected from H, methyl, and ethyl.

In some embodiments, Z is

In some embodiments, R⁶ is selected from In some embodiments, R⁶ is selected from wherein R⁷ is H, methyl optionally substituted with cyclopropyl, ethyl, propyl, iso-propyl, cyclopropyl, tetrahydrofuranyl, phenyl optionally substituted with one or more groups selected from methyl, ethyl, F, Cl, and CF₃, pyrrolidinyl optionally substituted with one or more groups selected from methyl, ethyl, F, Cl, and CF₃ provided that pyrrolidinyl is attached to the carbonyl carbon or sulfone sulfur via carbon atom, or tetrahydro-furanyl or pyranyl optionally substituted with one or more groups selected from methyl, ethyl, F, Cl, and CF₃, R⁸ is H, methyl, or ethyl, R⁹ is H, methyl, or ethyl; or R⁷ and R⁹, together with the nitrogen to which they are attached, form a pyrrolidine, piperazine, piperidine or morpholine each of which is optionally substituted with one or more groups selected from methyl, ethyl, F, Cl, and CF_3.

In some embodiments, R⁶ is selected from

In some embodiments, the compound or a pharmaceutically acceptable salt thereof as disclosed herein is selected from or a pharmaceutically acceptable salt thereof.

Also provided is a pharmaceutical composition comprising a compound or a pharmaceutically acceptable salt thereof as disclosed herein and a pharmaceutically acceptable excipient.

Also provided is method of inhibiting KRAS G12D activity in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of a composition as disclosed herein.

Also provided is a method of treating a disease or disorder associated with KRAS G12D mutation in a subject in need thereof, comprising administering to the patient a therapeutically effective amount of a composition as disclosed herein.

In some embodiments, the disease or disorder associated with KRAS G12D mutation is cancer. In some embodiments, the cancer is selected from carcinoma, squamous carcinoma, pancreatic cancer, prostate cancer, rectal cancer, colon cancer, colorectal cancer, non-small cell lung cancer, prostate cancer, small intestine cancer, sarcoma, leukemia, melanoma, and lymphoma.

In some embodiments, the method further comprises administering to the subject in need thereof an additional known anti-cancer agent.

DETAILED DESCRIPTION

I. Definitions

The substituents as disclosed herein intend to result in a chemical structure that is stable. Any substitution pattern that will render a compound known to be chemically unstable to a skilled artisan is not contemplated herein.

A dash ( "-" ) at the left hand side of a substituent is used to indicate a point of attachment for a substituent. For example, -CONH₂ is attached through the carbon atom.

The term "alkyl" herein refers to a straight or branched hydrocarbon chain containing 1-14 carbons. The symbol of C subscripted with a number range that precedes the term “alkyl” stands for the number of carbons in the alkyl. For example, C_1-5 alkyl represents an alkyl containing 1, 2, 3, 4, or 5 carbon atoms. Examples of C_1-5 alkyl groups include, but are not limited to, methyl, ethyl, n-propyl, i-propyl, n-butyl, i-butyl, t-butyl, and pentyl.

The term “alkenyl” herein refers to an unsaturated branched or straight hydrocarbon chain containing at least one carbon-carbon double bond. The group may be in either the cis or trans configuration about the double bond. The symbol of C subscripted with a number range that precedes the term “alkenyl” stands for the number of carbons in the alkenyl. For example, C_2-8 alkenyl represents an alkenyl containing 2, 3, 4, 5, 6, 7, or 8 carbon atoms. Exemplary alkenyl includes, but are not limited to, ethylenyl; propenyls such as prop-1-en-1-yl, prop-1-en-2-yl, prop-2-en-1-yl (allyl) , prop-2-en-2-yl, cycloprop-1-en-1-yl; cycloprop-2-en-1-yl; butenyls such as but-1-en-1-yl, but-1-en-2-yl, 2-methyl-prop-1-en-1-yl, but-2-en-1-yl, but-2-en-1-yl, but-2-en-2-yl, buta-1, 3-dien-1-yl, buta-1, 3-dien-2-yl, cyclobut-1-en-1-yl, cyclobut-1-en-3-yl, cyclobuta-1, 3-dien-1-yl; and the like. In certain embodiments, an alkenyl group has from 2 to 10 carbon atoms and in other embodiments, from 2 to 6 carbon atoms containing one carbon-carbon double bond.

The term “alkynyl” herein refers to an unsaturated branched or straight hydrocarbon chain containing at least one carbon-carbon triple bond. The symbol of C subscripted with a number range that precedes the term “alkynyl” stands for the number of carbons in the alkynyl. For example, C_2-8 alkynyl represents an alkynyl containing 2, 3, 4, 5, 6, 7, or 8 carbon atoms. Exemplary alkynylincludes, but are not limited to, ethynyl; propynyls such as prop-1-yn-1-yl, prop-2-yn-1-yl; butynyls such as but-1-yn-1-yl, but-1-yn-3-yl, but-3-yn-1-yl; and the like. In certain embodiments, an alkynyl group has from 2 to 10 carbon atoms and in other embodiments, from 2 to 6 carbon atoms containing one carbon-carbon triple bond.

The term “alkoxyl” or “alkoxy” herein refers to -O-alkyl. The symbol of C subscripted with a number range that precedes the term “alkoxy” stands for the number of carbons in the alkoxy. For example, C_1-5 alkoxy represents an alkoxy containing 1, 2, 3, 4, or 5 carbon atoms. Examples of C_1-5 alkoxy groups include, but are not limited to, methoxy, ethoxy, propyloxy, butoxy, and pentoxy.

The term “aryl” refers to a 6-10 ring membered monocyclic aromatic hydrocarbon ring, such as phenyl. Aryl also refers to a 8-14 ring membered spiro, fused, or bridged bi-, or multi-cyclic ring system, wherein at least one of the cyclics or rings is aromatic and does not comprise a heteroatom selected from O, S, and N as ring atom, the remaining cyclic (s) or ring (s) may be saturated, partially saturated, or aromatic, provided (1) when the remaining cyclic (s) or ring (s) is aromatic, it does not comprise a heteroatom selected from O, S, and N as ring atom, and (2) when the remaining cyclic (s) or ring (s) is not aromatic, it may or may not comprise a heteroatom selected from O, S, and N as ring atom. The point of attachment can be any ring atom. For example, are aryls.

The term "cycloalkyl" herein refers to a 3-14 ring membered saturated or partially unsaturated mono-cyclic, or spiro, fused, or bridged bi-, or multi-cyclic hydrocarbon group only having carbon atom as the ring atom. The symbol of C subscripted with a number range that precedes the term “cycloalkyl” stands for the carbon ring numbers in the cycloalkyl. For example, C_3-5 cycloalkyl represents a cycloalkyl containing 3, 4, or 5 carbon ring atoms, i.e., cyclopropyl, cyclobutyl, or cyclopentyl. The ring may be saturated or have one or more double bonds (i.e. partially unsaturated) , but not fully conjugated. When cycloalkyl is spiro, fused, or bridged bi-, or multi-cyclic, none of the cycles or rings is aromatic.

The term "heteroaryl" refers to 5-14 ring membered, such as 5 or 6 ring membered, mono-cyclic aromatic ring containing one or more, for example, from 1 to 4, or, in some embodiments, from 1 to 3, heteroatoms selected from N, O, and S, with the remaining ring atoms being carbon. Heteroaryl also refers to 7-14 ring membered spiro, fused, or bridged bi-, or multi-cyclic ring system, wherein at least one of the cyclics or rings is aromatic containing one or more, for example, from 1 to 4, or, in some embodiments, from 1 to 3, heteroatoms selected from N, O, and S as ring atoms, the remaining cyclic (s) or ring (s) (1) may or may not contain heteroatoms selected from N, O, and S and (2) may be saturated, partially saturated, or aromatic. The point of the attachment can be any ring atom. For example, are heteroaryls.

Further exemplary heteroaryl include, but are not limited to, pyridinyl, pyrazinyl, pyrazinyl, pyrimidinyl, pyrazolyl, imidazolinyl, isoxazolyl, oxazolyl, thiazolyl, thiadiazolyl, tetrazolyl, thienyl, benzothienyl, furyl, benzofuryl, benzoimidazolinyl, indolinyl, pyridizinyl, triazolyl, quinolinyl, pyrazolyl, and 5, 6, 7, 8-tetrahydroisoquinoline.

The term “heterocyclyl" herein refers to a 5 to 14 ring membered, saturated or partially unsaturated mono-cyclic ring, or fused, spiro, or bridged bicyclic or multicyclic ring, containing one or more, for example, from 1 to 4, or, in some embodiments, from 1 to 3, heteroatoms selected from N, O, and S, with the remaining ring atoms being carbon. The point of the attachment can be any ring atom. When heterocyclyl is spiro, fused, or bridged bi-, or multi-cyclic, none of the cycles or rings is aromatic.

Exemplary heterocyclyl includes but are not limited to pyrrolidinyl, piperidinyl, morpholinyl, piperazinyl, tetrahydro-furanyl, 5, 6, 7, 8-tetrahydroimidazo [1, 2-a] pyrazinyl, tetrahydro-2H-pyranyl, 8-oxa-3-azabicyclo [3.2.1] octanyl, 3-oxa-9-azaspiro [5.5] undecanyl, 7-oxa-2-azaspiro [3.5] nonanyl, and 2-oxa-7-azaspiro [3.5] nonanyl, azepanyl, 1, 2, 5-triazepanyl,6, 7, 8, 9-tetrahydro-1H, 5H- [1, 2, 4] triazolo [1, 2-a] [1, 2, 5] triazepinyl, diazepanyl, 1, 2, 5-oxadiazepanyl.

“Halo” refers to F. Cl, Br or I.

“Pharmaceutically acceptable salt” refers to a salt form of a compound (e.g., a drug) having at least one group capable for salt formation that causes no significant adverse toxicological effects to the subject. Pharmaceutically acceptable salts include, for example, salts prepared by reaction with an inorganic acid, organic acid, or a base depending on the nature of the compound (e.g., drug) . The inorganic acid can be hydrochloric acid, hydrobromic acid, carbonic acid, sulfuric acid, phosphoric acid, and the like; the organic acid can be fumaric acid, maleic acid, succinic acid, acetic acid, citric acid, tartaric acid, methanesulfonic acid and the like. The base that can form a salt with an acid drug can be an amine containing compound or inorganic base such as sodium hydroxide, sodium carbonate, and the like. Suitable pharmaceutically acceptable salt forms can e found in, for example, Handbook of Pharmaceutical Salts: Properties, Selection and Use, Weinheim/Zürich: Wiley-VCH/VHCA, 2002; P.H. Stahl and C.G. Wermuth, Eds.

The terms “treating” , “treatment” , or “treat” (of) a disease refers to slowing or arresting the development of a disease, providing relief from the symptoms or side-effects of the disease, and/or causing regression of the disease. The terms also refer to reduction of the occurrence of the disease in the subject when compared with a subject without the treatment.

A "pharmaceutically acceptable excipient" means an excipient that is useful in preparing a pharmaceutical composition that is generally safe, non-toxic and neither biologically nor otherwise undesirable, and includes a carrier or an excipient that is acceptable for veterinary use as well as human pharmaceutical use. "A pharmaceutically acceptable excipient" as used in the specification and claims includes both one and more than one such excipient.

The term “subject” as used herein refers to animal (such as mammal) or human.

Compound of formula (I) or a pharmaceutically acceptable salt thereof as described herein include, but are not limited to, their solvates, optical isomers, racemates, and other mixtures thereof. In those situations, the single enantiomers or diastereomers, i.e., optically active forms, can be obtained by asymmetric synthesis or by resolution of the racemates or mixtures of diastereomers. Resolution of the racemates or mixtures of diastereomers can be accomplished, for example, by conventional methods such as crystallization in the presence of a resolving agent, or chromatography, using, for example a chiral high-pressure liquid chromatography (HPLC) column. Where compounds described herein exist in various tautomeric forms, the term "compound" is intended to include all tautomeric forms of the compound. Moreover, compound of formula (I) or a pharmaceutically acceptable salt thereof as described herein also include compounds of formula (I) or a pharmaceutically acceptable salt thereof wherein certain atoms in formula (I) are replaced with their corresponding isotopes, such as certain H is replaced by D (deuterium) .

II. Compounds and uses thereof

Compounds disclosed herein (the term “compound (s) disclosed herein” includes pharmaceutically acceptable salt thereof) will be administered in a therapeutically effective amount by any of the accepted administration modes for agents in the form of a pharmaceutical composition that serve similar utilities. Therapeutically effective amount of the compounds disclosed herein may range from 0.01 to 500 mg per kg subject body weight, which can be administered in single or multiple doses per day. For oral administration, the pharmaceutical compositions can be provided in the form of tablets or capsules containing 1.0 to 1000 mg of the compound disclosed herein, such as, 1.0, 5.0, 10, 15, 20, 25, 50, 75, 100, 150, 200, 250, 300, 350, 400, 450, 500, 550, 600, 650, 700, 750, 800, 850, 900, and 1000 mg of the compound disclosed herein.

In addition to oral administration, the compound disclosed herein can also be administered as pharmaceutical compositions by, for example, transdermal, intranasal, suppository, intramuscular, intravenous or subcutaneous administration.

Thus, also provided is a pharmaceutical composition comprising the compound disclosed herein and a pharmaceutically acceptable excipient. When prepared for unit dosage form, the pharmaceutical compositions can comprise from 1 mg to 1000 mg of the compound disclosed herein.

Exemplary solid pharmaceutical excipient includes starch, cellulose, talc, glucose, lactose, sucrose, gelatin, malt, rice, flour, chalk, silica gel, magnesium stearate, sodium stearate, glycerol monostearate, sodium chloride, dried skim milk and the like. Liquid and semisolid excipients may be selected from glycerol, propylene glycol, water, ethanol and various oils, including those of petroleum, animal, vegetable or synthetic origin, e g, peanut oil, soybean oil, mineral oil, sesame oil, etc. Preferred liquid excipients, particularly for injectable solutions, include water, saline, aqueous dextrose, and glycols.

Other suitable pharmaceutical excipients and their formulations are described in Remington's Pharmaceutical Sciences, edited by E. W. Martin (Mack Publishing Company, 20th ed., 2000) .

Further provided is a method of inhibiting KRAS G12D activity in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of the compound described herein.

Further provided is a method of treating a disease or disorder associated with KRAS G12D mutation in a subject in need thereof, comprising administering to the patient a therapeutically effective amount of the compound described herein.

The disease or disorder associated with KRAS G12D mutation can be cancer. The cancer includes but not limited to carcinoma, squamous carcinoma, pancreatic cancer, prostate cancer, rectal cancer, colon cancer, colorectal cancer, non-small cell lung cancer, prostate cancer, small intestine cancer, sarcoma, leukemia, melanoma, and lymphoma.

The compound disclosed herein may be administered in combination with other anti-cancer agents, or in combination with radiation therapy or surgery. Other anti-cancer agents can be Paclitaxel, cisplatin, carboplatin and oxaliplatin, PARP inhibitor (such as niraparib, Olaparib) , anti-PD-1 antibody, anti-PD-L1 antibody, anti-CTLA-4 antibody, mTOR inhibitor, IGF1R inhibitor, HADC inhibitor, EGFR inhibitor, for example, anti-EGFR antibody (such as panitumumab) , HIF-1 inhibitor, VEGF/VEGFR inhibitors (such as sorafenib, bevacizumab) .

EXAMPLES

The following examples are illustrative in nature and are in no way intended to be limiting.

Example 1: Synthesis of N- ( (R) -1- (7- (8-ethyl-7-fluoro-3-hydroxynaphthalen-1-yl) -8-fluoro-2- ( ( (2R, 7aS) -2-fluorotetrahydro-1H-pyrrolizin-7a (5H) -yl) methoxy) pyrido [4, 3-d] pyrimidin-4-yl) piperidin-3-yl) acetamide (Compound 1)

Step 1: Synthesis of 2- (8-ethynyl-7-fluoro-3- (methoxymethoxy) naphthalen-1-yl) -4, 4, 5, 5-tetramethyl-1, 3, 2-dioxaborolane

To a solution of ( (2-fluoro-6- (methoxymethoxy) -8- (4, 4, 5, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) naphthalen-1-yl) ethynyl) triisopropylsilane (1.0 g, 1.95 mmol) in DMF (5 mL) was added CsF (3.0 g, 19.86 mmol) . The mixture was stirred for 1 h at 50℃. The reaction was allowed to cool down to room temperature and poured into H₂O (100 mL) . Large amount of white solid precipitated out. It was stirred for another 10 min, filtered, and the cake was washed with H₂O. The cake was collected and dried to afford the title compound (1-1) (650 mg, 93%yield) .

Step 2: Synthesis of 2- (8-ethyl-7-fluoro-3- (methoxymethoxy) naphthalen-1-yl) -4, 4, 5, 5-tetramethyl-1, 3, 2-dioxaborolane

To a solution of 2- (8-ethynyl-7-fluoro-3- (methoxymethoxy) naphthalen-1-yl) -4, 4, 5, 5-tetramethyl-1, 3, 2-dioxaborolane (650 mg, 1.82 mmol) in MeOH (50 mL) was added 10%Pd/C (100 mg) . The mixture was stirred under H₂ atmospherefor 1 h at 35 ℃. The reaction mixture was allowed to cool down to room temperature, filtered through a short pad of celite, and washed with MeOH (50 mL) . The filtrate was concentrated to afford the title compound (1-2) (550 mg, 83%yield) .

Step 3: Synthesis of 2, 4, 7-trichloro-8-fluoropyrido [4, 3-d] pyrimidine

To a solution of 7-chloro-8-fluoropyrido [4, 3-d] pyrimidine-2, 4 (1H, 3H) -dione (1-4) (20.0 g, 93.4 mmol) in phosphorus chloride (200 mL) at 0 ℃ was added N, N-diisopropylethylamine (76 mL, 468 mmol) . The reaction solution was heated at 110 ℃ for ～2 hours till completion determined by TLC. The reaction liquid was cooled to room temperature and concentrated under vacuum to remove most of the phosphorus oxychloride. The resulting mixture was quenched with ice water (120 mL) , and then adjusted the pH to neutral with saturated aqueous sodium bicarbonate solution. The mixture was extracted with ethyl acetate (150 mL x2) . The combined organics were washed with brine (150 mL) , dried over anhydrous sodium sulfate dried, and concentrated. The crude product was purified by silica gel column chromatography (petroleum ether/ethyl acetate = 3/1) to provide the yellow solid compound 2, 4, 7-trichloro-8-fluoropyrido [4, 3-d] pyrimidine (1-5) (13.6 g, 58%yield) .

Step 4: Synthesis of 4- (benzyloxy) -2, 7-dichloro-8-fluoropyrido [4, 3-d] pyrimidine

To a solution of 2, 4, 7-trichloro-8-fluoropyrido [4, 3-d] pyrimidine (1-5) (10.7 g, 42.38 mmol) and N, N-diisopropylethylamine (16.41 g, 127.15 mmol) in dioxane (50 mL) at room temperature was added benzyl alcohol (5.04 g, 46.62 mmol) . The reaction solution was slowly heated to 60 ℃ and stirred for ～3 hours till completion determined by TLC. The reaction solution was cooled to room temperature, quenched with water (100 mL) , and extracted with ethyl acetate (150 mL x2) . The combined organics were dried by anhydrous sodium sulfate, and concentrated. The crude product was purified by silica gel column chromatography (petroleum ether/ethyl acetate = 6%-20%) to provide a white solid compound 4- (benzyloxy) -2, 7-dichloro-8-fluoropyrido [4, 3-d] pyrimidine (1-6) (9.7 g, 70%yield) .

Step 5: Synthesis of 4- (benzyloxy) -7-chloro-8-fluoro-2- ( ( (2R, 7aS) -2-fluorotetrahydro-1H-pyrrolizin-7a (5H) -yl) methoxy) pyrido [4, 3-d] pyrimidine

A mixture of 4- (benzyloxy) -2, 7-dichloro-8-fluoropyrido [4, 3-d] pyrimidine (1-6) (5.1 g, 15.74 mol) , cesium carbonate (12.79 g, 39.35 mmol) , and ( (2R, 7aS) -2-fluorotetrahydro-1H-pyrrolizin-7a (5H) -yl) methanol (1-7) (3.51 g, 22.07 mmol) in anhydrous dioxane (75 mL) was heated at 80 ℃ under nitrogen protection for ～6 hours till completion determined by TLC. The reaction solution was then concentrated under vacuum, quenched with water, and extracted with ethyl acetate (100 mL x3) . The combined organics were dried over anhydrous sodium sulfate, and concentrated. The crude product was purified by silica gel column chromatography (methanol/dichloromethane = 0-10%) to obtain a white solid compound 4- (benzyloxy) -7-chloro-8-fluoro-2- ( ( (2R, 7aS) -2-fluorotetrahydro-1H-pyrrolizin-7a (5H) -yl) methoxy) pyrido [4, 3-d] pyrimidine (1-8) (3.8 g, 54%yield) . LC-MS: m/z = 447.2 [M+H] ⁺.

Step 6: Synthesis of 4- (benzyloxy) -7- (8-ethyl-7-fluoro-3- (methoxymethoxy) naphthalen-1-yl) -8-fluoro-2- ( ( (2R, 7aS) -2-fluorotetrahydro-1H-pyrrolizin-7a (5H) -yl) methoxy) pyrido [4, 3-d] pyrimidine

A mixture of 4- (benzyloxy) -7-chloro-8-fluoro-2- ( ( (2R, 7aS) -2-fluorotetrahydro-1H-pyrrolizin-7a (5H) -yl) methoxy) pyrido [4, 3-d] pyrimidine (1-8) (300 mg, 0.67 mmol) , 2- (8-ethyl-7-fluoro-3- (methoxymethoxy) naphthalen-1-yl) -4, 4, 5, 5-tetramethyl-1, 3, 2-dioxaborolane (1-3) (500 mg, 1.38 mmol) , K₃PO₄ (456 mg, 2.15 mmol) , (2-dicyclohexylphosphino-2′, 6′-diisopropoxy-1, 1′-biphenyl) [2- (2′-amino-1, 1′-biphenyl) ] palladium (II) methanesulfonate (104 mg, 0.14 mmol) in 1, 4-dioxane (6 mL) and H₂O (2 mL) was heated under N₂ atmosphere for 1.5 h at 90℃. The reaction mixture was allowed to cool down to room temperature, diluted with H₂O (100 mL) , and extracted with EtOAc (3 x 200 mL) . The combined organic phases were washed with brine, dried over anhydrous Na₂SO₄, and filtered. The filtrate was concentrated to afford the crude title compound 1-9as yellow oil (450 mg) , which was used for next step without further purification. LC-MS: m/z=645.3 [M+H] ⁺

Step 7: Synthesis of 7- (8-ethyl-7-fluoro-3- (methoxymethoxy) naphthalen-1-yl) -8-fluoro-2- ( ( (2R, 7aS) -2-fluorotetrahydro-1H-pyrrolizin-7a (5H) -yl) methoxy) pyrido [4, 3-d] pyrimidin-4-ol

To a solution of above crude product 1-9in EtOAc (50 mL) was added 10%Pd/C (100 mg) . The mixture was stirred under H₂ atmosphere for 2 hrs at 35 ℃. The reaction mixture was allowed to cool down to room temperature and filtered through a short pad of celite and washed with MeOH (50 mL) . The filtrate was concentrated and the residue was purified by silica gel chromatography, eluted with gradian 0-10%MeOH/DCM to afford the title compound (1-10) (190 mg, yield 51%for two steps) . LC-MS: m/z=555.2 [M+H] ⁺

Step8: Synthesis ofN- ( (R) -1- (7- (8-ethyl-7-fluoro-3- (methoxymethoxy) naphthalen-1-yl) -8-fluoro-2- ( ( (2R, 7aS) -2-fluorotetrahydro-1H-pyrrolizin-7a (5H) -yl) methoxy) pyrido [4, 3-d] pyrimidin-4-yl) piperidin-3-yl) acetamide

To a solution of 7- (8-ethyl-7-fluoro-3- (methoxymethoxy) naphthalen-1-yl) -8-fluoro-2- ( ( (2R, 7aS) -2-fluorotetrahydro-1H-pyrrolizin-7a (5H) -yl) methoxy) pyrido [4, 3-d] pyrimidin-4-ol (1-10) (155 mg, 0.28 mmol) in DMA (10 mL) was added DIPEA (616 mg, 1.62 mmol) , HATU (410 mg, 1.08 mmol) and (R) -N- (piperidin-3-yl) acetamide (1-11) (235 mg, 1.65 mmol) . The mixture was stirred for 12 h at 30℃ before it was quenched by H₂O (100 mL) . The resulting mixture was extracted with EtOAc (2 x 100 mL) . The combined organic layers were washed with brine, dried over anhydrous Na₂SO₄ and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by silica gel chromatography and eluted with DCM/MeOH (20/1) to afford titled compound (1-12) as a white solid (125 mg, 68%yield) . LC-MS: m/z = 679.3 [M+H] ⁺ _.

Step9: Synthesis ofN- ( (R) -1- (7- (8-ethyl-7-fluoro-3-hydroxynaphthalen-1-yl) -8-fluoro-2- ( ( (2R, 7aS) -2-fluorotetrahydro-1H-pyrrolizin-7a (5H) -yl) methoxy) pyrido [4, 3-d] pyrimidin-4-yl) piperidin-3-yl) acetamide

To a solution of N- ( (R) -1- (7- (8-ethyl-7-fluoro-3- (methoxymethoxy) naphthalen-1-yl) -8-fluoro-2- ( ( (2R, 7aS) -2-fluorotetrahydro-1H-pyrrolizin-7a (5H) -yl) methoxy) pyrido [4, 3-d] pyrimidin-4-yl) piperidin-3-yl) acetamide (1-12) (125 mg, 0.184 mmol) in DCM (3 mL) was added TFA (3 mL) . After stirring for 2 hrs at room temperature, the reaction mixture was diluted with H₂O (10 mL) and extracted with DCM (2 x 10 mL) . The combined organic layers were washed with brine, dried over anhydrous Na₂SO₄ and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by prepared TLC to afford the titled compound (1) as a white solid (80 mg, 68%yield) . LC-MS: m/z = 635.3 [M+H] ⁺; ¹H NMR (400 MHz, CD₃OD) δ 9.15 (d, J = 2.8 Hz, 1H) , 7.71-7.63 (m, 1H) , 7.30 (d, J = 2.4 Hz, 1H) , 7.25 (t, J = 9.2 Hz, 1H) , 7.05 (t, J = 2.8 Hz, 1H) , 5.46-5.31 (m, 1H) , 4.57-4.45 (m, 2H) , 4.42-4.30 (m, 2H) , 4.12-4.03 (m, 1H) , 3.76-3.66 (m, 1H) , 3.61-3.47 (m, 2H) , 3.44-3.36 (m, 2H) , 3.17-3.10 (m, 1H) , 2.51-1.93 (m, 13H) , 1.88-1.72 (m, 2H) , 0.87-0.73 (m, 3H) ; ¹⁹FNMR (377 MHz, CD₃OD) δ -121.18, -139.19, -173.75.

Example 2: Synthesis of methyl ( (R) -1- (7- (8-ethynyl-7-fluoro-3-hydroxynaphthalen-1-yl) -8-fluoro-2- ( ( (2R, 7aS) -2-fluorotetrahydro-1H-pyrrolizin-7a (5H) -yl) methoxy) pyrido [4, 3-d] pyrimidin-4-yl) piperidin-3-yl) carbamate (Compound 2)

Step 1: Synthesis of8-fluoro-7- (7-fluoro-3- (methoxymethoxy) -8- ( (triisopropylsilyl) ethynyl) naphthalen-1-yl) -2- ( ( (2R, 7aS) -2-fluorotetrahydro-1H-pyrrolizin-7a (5H) -yl) methoxy) pyrido [4, 3-d] pyrimidin-4-ol

A mixture of 4- (benzyloxy) -7-chloro-8-fluoro-2- ( ( (2R, 7aS) -2-fluorotetrahydro-1H-pyrrolizin-7a (5H) -yl) methoxy) pyrido [4, 3-d] pyrimidine (1-8) (3.0 g, 6.72 mmol) , ( (2-fluoro-6- (methoxymethoxy) -8- (4, 4, 5, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) naphthalen-1-yl) ethynyl) triisopropylsilane (1-1) (4.13 g, 8.06 mmol) , Cs₂CO₃ (5.47 g, 16.8 mmol) and (2-dicyclohexylphosphino-2′, 6′-diisopropoxy-1, 1′-biphenyl) [2- (2′-amino-1, 1′-biphenyl) ] palladium (II) methanesulfonate (0.84 g, 1.01 mmol) in EtOH (60 mL) and H₂O (15 mL) was stirred under N₂ atmosphere at 100℃ for 2.5 hrs. The reaction mixture was cool down to room temperature, diluted with H₂O (100 mL) , extracted with EtOAc (2 x 200 mL) . The combined organic layers were washed with brine (200 mL) , dried over anhydrous Na₂SO₄, and filtered. The filtrated was concentrated andthe residue was purified by silica gel chromatography with gradient elution DCM/MeOH 50/1～20/1) to affordthe title compound (2-1) as a yellow solid (2.24 g, 47%yield) . LC-MS: m/z=707.3 [M+H] ⁺ _.

Step 2: Synthesis of7- (8-ethynyl-7-fluoro-3- (methoxymethoxy) naphthalen-1-yl) -8-fluoro-2- ( ( (2R, 7aS) -2-fluorotetrahydro-1H-pyrrolizin-7a (5H) -yl) methoxy) pyrido [4, 3-d] pyrimidin-4-ol

A mixture of 8-fluoro-7- (7-fluoro-3- (methoxymethoxy) -8- ( (triisopropylsilyl) ethynyl) naphthalen-1-yl) -2- ( ( (2R, 7aS) -2-fluorotetrahydro-1H-pyrrolizin-7a (5H) -yl) methoxy) pyrido [4, 3-d] pyrimidin-4-ol (2-1) (2.2 g, 3.11 mmol) and CsF (9.5 g, 62.2 mmol) in DMF (20 mL) was stirred 3 hrs at room temperature. The reaction mixture was dilute with H₂O (100 mL) , extracted with EtOAc (2 x 100 mL) . The combined organic layers were washed with brine, dried over anhydrous Na₂SO₄, and filtered. The filtrate was concentrated andthe residue was stirred with EtOAc (30 mL) . The resulting solid was collected via filtration and dried to afford the title compound (2-3) as a white solid (1.35 g, 79%yield) . LC-MS: m/z=551.2 [M+H] ⁺; ¹H NMR (400 MHz, DMSO-d₆) δ 8.93 (s, 1H) , 8.10 (dd, J = 9.2, 6.0 Hz, 1H) , 7.74 (d, J = 2.4 Hz, 1H) , 7.55 (t, J = 8.8 Hz, 1H) , 7.36 (d, J = 2.4 Hz, 1H) , 5.38 (s, 2H) , 4.25-4.07 (m, 3H) , 3.44 (s, 3H) , 3.24-2.98 (m, 4H) , 2.91-2.76 (m, 1H) , 2.23-1.99 (m, 3H) , 1.88-1.73 (m, 3H) .

Step 3: Synthesis ofmethyl ( (R) -1- (7- (8-ethynyl-7-fluoro-3- (methoxymethoxy) naphthalen-1-yl) -8-fluoro-2- ( ( (2R, 7aS) -2-fluorotetrahydro-1H-pyrrolizin-7a (5H) -yl) methoxy) pyrido [4, 3-d] pyrimidin-4-yl) piperidin-3-yl) carbamate

The title compound (2-4) (90 mg, 66%yield) was prepared following the procedure described in Example 1 step 8, using 7- (8-ethynyl-7-fluoro-3- (methoxymethoxy) naphthalen-1-yl) -8-fluoro-2- ( ( (2R, 7aS) -2-fluorotetrahydro-1H-pyrrolizin-7a (5H) -yl) methoxy) pyrido [4, 3-d] pyrimidin-4-ol (2-2) (100 mg, 0.197 mmol) to replace (1-10) , methyl (R) -piperidin-3-ylcarbamate TFA salt (2-3) to replace (1-11) . LC-MS: m/z=691.3 [M+H] ⁺.

Step 4: Synthesis ofmethyl ( (R) -1- (7- (8-ethynyl-7-fluoro-3-hydroxynaphthalen-1-yl) -8-fluoro-2- ( ( (2R, 7aS) -2-fluorotetrahydro-1H-pyrrolizin-7a (5H) -yl) methoxy) pyrido [4, 3-d] pyrimidin-4-yl) piperidin-3-yl) carbamate

Methyl ( (R) -1- (7- (8-ethynyl-7-fluoro-3- (methoxymethoxy) naphthalen-1-yl) -8-fluoro-2- ( ( (2R, 7aS) -2-fluorotetrahydro-1H-pyrrolizin-7a (5H) -yl) methoxy) pyrido [4, 3-d] pyrimidin-4-yl) piperidin-3-yl) carbamate (2-4) (90 mg, 0.13 mmol) was dissolved in dioxane (5 mL) and TFA (10 mL) . The mixture was stirred at room temperature for 4 hrs. It was neutralized with saturated NaHCO₃ aqueous solution to pH = 8-9, and thenextracted with EtOAc (2 x 20 mL) . The combined organic phases were washed with H₂O (20 mL) , brine (10 mL) , dried over anhydrous Na₂SO₄, and filtered. The filtrate was concentrated andthe residue was purified by silica gel chromatography to afford the title Compound 2 as a white solid (35mg, 41.6%yield) . LC-MS: m/z = 647.3 [M+H] ⁺; ¹H NMR (400 MHz, CD₃OD) δ 9.10 (d, J = 8.0 Hz, 1H) , 7.88-7.84 (m, 1H) , 7.36-7.30 (m, 2H) , 7.23 (dd, J = 1.6, 2.4 Hz, 1H) , 5.52-5.39 (m, 1H) , 4.72-4.63 (m, 2H) , 4.47-4.43 (m, 2H) , 3.88-3.53 (s, 8H) , 3.48-3.36 (m, 2H) , 3.29-3.21 (m, 1H) , 2.60-2.28 (m, 3H) , 2.23-1.92 (m, 5H) , 1.83-1.66 (m, 2H) ; ¹⁹F NMR (377 MHz, CD₃OD) δ -111.66, -140.05, -173.83.

The following compounds weresynthesized using similar procedure as described above for Example 1.

The following compounds were synthesized using similar procedure as described above for Example 2.

The following compounds can be synthesized using similar chemistry and procedure as described above for Example 1 and 2.

pharmaceutically acceptable salt thereof.

Example 3: In vitro fluorescence polarization (FP) binding assay

Assay Principle: after the fluorescently labeled cyclic peptide ligand binds to KRAS G12D protein, when excited by polarized light, the fluorescent molecule remains stationary, and will emit light with a fixed polarization plane (the emission light still maintains polarization) . In contrast to the fast rotation or flipping of the cyclic peptide molecules in the unbound state, the emitted light will be depolarized relative to the plane of the excitation light. The compound competes for the binding site of the ligand to KRAS G12D, and the binding efficiency of the compound to KRAS G12D is obtained by detecting the change in the polarization degree of the emitted light.

Assay Procedures: prepared1x reaction buffer (50 mM Tris 7.5 Sinopharm, Cat. No. 30188360) . KRAS G12D (Purified in house) was added to 1x reaction buffer to make a 3x enzyme solution (final concentration of KRAS G12D was 25 nM) , and 20 μL of this solution was transferred to different concentrations of compound (1%final concentration in DMSO) placed in 384-well microplates (Corning 384-well Polystyrene Microplates, Cat. No. 3575) . The microplates were incubated at room temperature for 15 minutes. The cyclic peptide ligand that specifically binds to KRAS G12D was added to 1x reaction buffer to prepare a 3x substrate solution, then 20 μL of substrate solution (final concentration of cyclic peptide ligand 5nM) was added to the wells of the microplates. The reaction was carried out at room temperature for a period of time. Then, the microplates were placed on the EnVision instrument to read the mp. After obtaining the mp raw data of different concentrations of the compound, the data were standardized according to the formula Inh%=100- (Sample-Min) / (Max-Min) *100%to obtain the enzyme activity inhibition rate Inh%of each concentration point (wherein Max is the mp value of the enzyme-positive wells, Min is the mp value of the enzyme-free negative wells, and Sample is the mp value of the compound-treated sample wells) , and then the inhibition rate Inh%(Y) corresponding to each concentration (X) was entered in EXCEL, andGraphpad plugin was used to calculate the half-inhibitory concentration IC₅₀ value of each compound according to the built-in four-parameter fitting formula Y=Bottom + (Top-Bottom) / (1+ (IC₅₀/X) *HillSlope) .

Example 4: Cell Based p-ERK Assay

This Example illustrates that exemplary compounds disclosed herein inhibit the intracellular phosphorylation of ERK downstream of KRAS G12D.

This experiment directly tested the inhibition of compounds on KRAS G12D at the cellular level by detesting the endogenous phosphorylation level of ERK1/2. After activation of

antibodies (one labeled with Eu3+-Cryptate (donor) and the other with D2 (acceptor) to recognize phosphorylated ERK1/2 (Thr202/Tyr204) site and the ERK1/2 protein itself, respectively. When phosphorylation occurs, the two dyes are approached by a light source (laser or flashlight) to excite fluorescence resonance energy transfer (FRET) from the donor to the acceptor, which emits a fluorescence wavelength (665 nm) at a specific location. The specific signal is proportional to the level of phospho-ERK1/2 (Thr202/Tyr204) .

Procedure:

GP5d cells expressing KRAS G12D mutation (ECACC, Cat. No. 95090715) were cultured in DMEM medium containing 10%FBS. Seed the cells in the 3D assay plate and cultured at 37℃ for 3 days. Treated with a series of compounds at a final concentration of 0.5%DMSO. After incubation for 2 hours, remove the supernatant, add 1*lysate to lyse the cells, transfer the lysate to a new assay plate, add the mixed antibody solution (Cisbio, Cat. No. 64AERPEH) was incubated overnight at room temperature. The microplate was placed on the EnVision instrument to read the Em665/620 fluorescence signal. According to the formula Inh %=100- (Sample-Min) / (Max-Min) *100%, the data is normalized to obtain the enzyme activity inhibition rate Inh %of each concentration point (wherein Max is the Em665/620 value containing enzyme-positive wells, Min is the Em665/620 value of the enzyme-free negative wells, Sample is the Em665/620 value of the compound-treated sample wells) , enter the inhibition rate Inh% (Y) corresponding to each concentration (X) in EXCEL, IC50 was calculated with Graphpad Prism based on the four-parameter fitting formula Y=Bottom + (Top-Bottom) / (1+ (IC50/X) *HillSlope)

The IC₅₀ data were listed in the table below: “+++” represents IC₅₀< 1μM, “++” represents 1 μM ≤IC₅₀< 5μM, “+” represents IC₅₀ ≥ 5 μM.

Claims

A compound of formula I or a pharmaceutically acceptable salt thereof:

wherein

Q¹, Q², and Q³ are independently selected from N, CH, C-CF₃, C-OH, C-Cl, C-F, C-CH₃, C-CH (CH₃) ₂, C-OCH₃, C-SCF₃, C-OCF₃, and C-CN;

Z is

ring A is a 4-10 membered heterocyclyl or a 8-10 membered fused heteroaryl, provided when ring A is a 8-10 membered fused heteroaryl, R⁶-L₁ is optionally present; n is 0, 1, or 2;

ring B is a 4, 5, 6, or 7 membered heterocycle and together with ring A forms a spiral bicyclic wherein ring B contains a moiety selected from ring B is further optionally substituted with one or more W;

R¹ is selected from L-5-12 membered heterocyclyl and L-C_3-8 cycloalkyl, wherein each of the 5-12 membered heterocyclyl and C_3-8 cycloalkyl is optionally substituted with one or more W;

L and L₁ are independently selected from -CH₂-, -CH (CH₃) -, CH₂-CH₂-, and a bond;

Y is selected from a bond and -O-;

R² is H or C_1-3 alkyl, R⁵ is C_1-6alkyl substituted with R⁶;

R⁶ is selected from

R³ is selected from aryl and heteroaryl, wherein the aryl and heteroaryl is optionally substituted with one or more W;

R⁴ is selected from H, -CF₃, -OH, OMe, OEt, -CH₃, F, and Cl;

R⁷ is selected from H, C_1-6 alkyl optionally substituted with one or more W, aryl optionally substituted with one or more W, cycloalkyl optionally substituted with one or more W, and 5, 6, or 7 membered heterocyclyl optionally substituted with one or more W, provided that the point of attachment of the heterocyclyl to the carbonyl carbon or sulfone sulfur is a carbon atom;

R⁸ is selected from H and C_1-6 alkyl; or R⁷ and R⁸, together with the atom (s) to which they are attached, form a 5, 6, or 7 membered heterocyclyl optionally substituted with one or more W;

R⁹ is selected from H and C_1-6 alkyl; or R⁷ and R⁹, together with the nitrogen to which they are attached, form a 5, 6, or 7 membered heterocyclyl optionally substituted with one or more W; and

W is selected from OH, CN, halo, C_2-4 alkynyl, NH₂, mono or di-C_1-6 alkyl substituted amino, C_2- ₄ alkenyl, C_3-8 cycloalkyl optionally substituted with 1-5 F or Cl, C_1-6 alkoxy optionally substituted with 1-5 F or Cl, and C_1-6 alkyl optionally substituted with one or more groups selected from F, Cl, -OC (O) NR^7’R^9’, and -NR^7’R^9’, with R^7’ and R^9’ independently being selected from H and C_1-6 alkyl, or R^7’ and R^9’, together with the nitrogen to which they are attached, form a 5, 6, or 7 membered heterocyclyl optionally substituted with one or more groups selected from F, Cl, C_1-6 alkyl..
The compound or a pharmaceutically acceptable salt thereof of claim 1, wherein Q¹ is N, one of Q² and Q³ is N, and the other is not.
The compound or a pharmaceutically acceptable salt thereof of claim 1, wherein Q¹ is N, Q² is selected from N, CH, C-CF₃, C-OH, C-Cl, C-F, C-CH₃, C-CH (CH₃) ₂, C-OCH₃, C-SCF₃, C-OCF₃, and C-CN, and Q³ is not N.
The compound or a pharmaceutically acceptable salt thereof of claim 1, wherein Q¹ is N, Q² is selected from N, CH, C-CF₃, C-OH, C-Cl, C-F, C-CH₃, C-CH (CH₃) ₂, C-OCH₃, C-SCF₃, C-OCF₃, and C-CN, and Q³ is C-H or C-F.
The compound or a pharmaceutically acceptable salt thereof of any of claims 1-4, wherein R³ is phenyl, naphthyl, pyridinyl, benzothiazolyl, benzothiophenyl, 1H-indenyl, 1H-indazolyl, or 1H-indolyl, each of which is optionally substituted with one or more W.
The compound or a pharmaceutically acceptable salt thereof of claim 5, wherein R³ is phenyl, naphthyl, pyridinyl, benzothiazolyl, benzothiophenyl, 1H-indenyl, 1H-indazolyl, or 1H-indolyl, each of which is optionally substituted with one or more groups selected from F, Cl, methyl, ethyl, ethenyl, ethynyl, CF₃, cyclopropyl, iso-propyl, -NH₂, -CN, and OH.
The compound or a pharmaceutically acceptable salt thereof of claim 6, wherein R³ is selected from
The compound or a pharmaceutically acceptable salt thereof of claim 7, wherein , R³ is selected from
The compound or a pharmaceutically acceptable salt thereof of any of claims 1-8, wherein Y is O.
The compound or a pharmaceutically acceptable salt thereof of any of claims 1-9, wherein R¹ is selected from -L-fused 7-10 membered bicyclic heterocyclyl comprising at least one nitrogen, and -L-5-7 membered monocyclic heterocyclyl comprising at least one nitrogen, each of the fused 7-10 membered bicyclic heterocyclyl comprising at least one nitrogen and 5-7 membered monocyclic heterocyclyl comprising at least one nitrogen is optionally substituted with one or more W.
The compound or a pharmaceutically acceptable salt thereof of claim 10, wherein each of the fused 7-10 membered bicyclic heterocyclyl comprising at least one nitrogen and 5-7 membered monocyclic heterocyclyl comprising at least one nitrogen is optionally substituted with one or more group selected from F and C_1-3 alkyl optionally substituted with -OC (O) NR^7’R^9’ or -NR^7’R^9’, with R^7’ and R^9’ independently being selected from H and C_1-6 alkyl, or R^7’ and R^9’, together with the nitrogen to which they are attached, form a 5, 6, or 7 membered heterocyclyl optionally substituted with one or more groups selected from F, Cl, C_1-6 alkyl.
The compound or a pharmaceutically acceptable salt thereof of claim 11, wherein R^7’ and R^9’ are methyl or R^7’ and R^9’, together with the nitrogen to which they are attached, form a morpholine.
The compound or a pharmaceutically acceptable salt thereof of any of claims 1-9, wherein R¹ is L-C_3-8 cycloalkyl optionally substituted with one or more W.
The compound or a pharmaceutically acceptable salt thereof of claim 13, wherein R¹ is L-C_3-8 cycloalkyl optionally substituted with methyl, wherein the methyl is optionally substituted with -OC (O) NR^7’R^9’ or -NR^7’R^9’, with R^7’ and R^9’ independently being selected from H and C_1-6 alkyl, or R^7’ and R^9’, together with the nitrogen to which they are attached, form a 5, 6, or 7 membered heterocyclyl optionally substituted with one or more groups selected from F, Cl, C_1-6 alkyl.
The compound or a pharmaceutically acceptable salt thereof of claim 14, wherein R¹is L-C_3-8 cycloalkyl optionally substituted with methyl, wherein the methyl is optionally substituted with -O (CO) N (CH₃) ₂, -O (CO) -morpholine, -N (CH₃) ₂, or morpholine.
The compound or a pharmaceutically acceptable salt thereof of any of claims1-9, wherein R¹ is
The compound or a pharmaceutically acceptable salt thereof of any of claims 1-16, wherein L is CH₂ or CH (CH₃) .
The compound or a pharmaceutically acceptable salt thereof of any of claims 1-17, wherein Z isin which ring A is a 4-10 membered heterocyclyl having only one atom on the ring is a heteroatom selected from N, O, and S, ring B, W, and n are as defined in claim 1.
The compound or a pharmaceutically acceptable salt thereof of claim 18, wherein ring A is a 4-10 membered heterocyclyl wherein the only one heteroatom on the ring is N.
The compound or a pharmaceutically acceptable salt thereof of claim 19, wherein ring A is a 4-10 membered heterocyclyl wherein the only one heteroatom on the ring is N and the N is the point of attachment as indicated by
The compound or a pharmaceutically acceptable salt thereof of claim 19, wherein ring A is
The compound or a pharmaceutically acceptable salt thereof of claim 21, wherein ring A is
The compound or a pharmaceutically acceptable salt thereof of claim 22, wherein ring A is
The compound or a pharmaceutically acceptable salt thereof of any of claims 18-23, wherein ring B is a 5 or 6 membered heterocycle containing a moiety selected from in which R⁸ and R⁹ are as defined in claim 1.
The compound or a pharmaceutically acceptable salt thereof of claim 24, wherein R⁸ and R⁹ are independently selected from H, methyl, and ethyl.
The compound or a pharmaceutically acceptable salt thereof of claim 24, wherein ring B is a 5 membered heterocycle containing wherein R⁸ and R⁹ are independently selected from H, methyl, and ethyl.
The compound or a pharmaceutically acceptable salt thereof of claim 24, wherein ring B is a 5 or 6 membered heterocycle containingwherein R⁸is selected from H, methyl, and ethyl.
The compound or a pharmaceutically acceptable salt thereof of any of claims 18-23, wherein ring B is a 4 membered heterocycle containing
The compound or a pharmaceutically acceptable salt thereof of claim 24, Z is
The compound or a pharmaceutically acceptable salt thereof of any of claims 18-29, wherein n is 0.
The compound or a pharmaceutically acceptable salt thereof of any of claims 18-29, wherein n is 1 and W is methyl or OH.
The compound or a pharmaceutically acceptable salt thereof of any of claims 1-17, wherein Z iswherein R² is H or C_1-3 alkyl, R⁵ is C_3-6 alkyl substituted with R⁶ as defined in claim 1.
The compound or a pharmaceutically acceptable salt thereof of claim 32, wherein R⁵ is propyl substituted with R⁶ as defined in claim 1.
The compound or a pharmaceutically acceptable salt thereof of claim 32, wherein Z isand R⁶ is as defined in claim 1.
The compound or a pharmaceutically acceptable salt thereof of any of claims 1-17, wherein Z iswherein ring A is a 4-10 membered heterocyclyl having only one atom on the ring is a heteroatom selected from N, O, and S or ring A is a 8-10 membered fused bi-cyclic heteroaryl with R⁶-L₁ being optionally present, L₁, R⁶, W, and n are as defined in claim 1.
The compound or a pharmaceutically acceptable salt thereof of claim 35, wherein ring A is a 4-10 membered heterocyclyl wherein the only one heteroatom on the ring is N.
The compound or a pharmaceutically acceptable salt thereof of claim 35, wherein ring A is a 4-10 membered heterocyclyl wherein the only one heteroatom on the ring is N and the N is the point of attachment as indicated by
The compound or a pharmaceutically acceptable salt thereof of claim 36, wherein ring A is
The compound or a pharmaceutically acceptable salt thereof of claim 38, wherein ring A is
The compound or a pharmaceutically acceptable salt thereof of claim 39, wherein ring A is
The compound or a pharmaceutically acceptable salt thereof of claim 35, wherein ring A is a 8-10 membered fused bi-cyclic heteroaryl wherein one ring is aromatic and the other is not aromatic.
The compound or a pharmaceutically acceptable salt thereof of claim 41, wherein ring A is a 8-10 membered fused bi-cyclic heteroaryl wherein one ring is aromatic, the other is not aromatic and is the point of attachment as indicated by
The compound or a pharmaceutically acceptable salt thereof of claim 42, wherein ring A is
The compound or a pharmaceutically acceptable salt thereof of claim 43, wherein ring A is
The compound or a pharmaceutically acceptable salt thereof of any of claims 35-44, wherein L₁ is a bond or CH₂.
The compound or a pharmaceutically acceptable salt thereof of any of claims 35-45, wherein n is 0.
The compound or a pharmaceutically acceptable salt thereof of any of claims 35-45, wherein n is 1 and W is selected from OH, methyl, and ethyl.
The compound or a pharmaceutically acceptable salt thereof of any of claims 32-47, wherein R⁶ is selected from wherein R⁷, R⁸, and R⁹ are as defined in claim 1.
The compound or a pharmaceutically acceptable salt thereof of any of claims 32-47 4, wherein R⁶ is selected from

R⁷ is H, methyl optionally substituted with cyclopropyl, ethyl, propyl, iso-propyl, cyclopropyl, tetrahydrofuranyl, phenyl optionally substituted with one or more groups selected from methyl, ethyl, F, Cl, and CF₃, pyrrolidinyl optionally substituted with one or more groups selected from methyl, ethyl, F, Cl, and CF₃ provided that pyrrolidinyl is attached to the carbonyl carbon or sulfone sulfur via carbon atom, or tetrahydro-furanyl or pyranyl optionally substituted with one or more groups selected from methyl, ethyl, F, Cl, and CF₃,

R⁸ is H, methyl, or ethyl,

R⁹ is H, methyl, or ethyl, or R⁷ and R⁹, together with the nitrogen to which they are attached, form a pyrrolidine, piperazine, piperidine or morpholine each of which is optionally substituted with one or more groups selected from methyl, ethyl, F, Cl, and CF₃.
The compound or a pharmaceutically acceptable salt thereof of claim 38, wherein R⁶ is selected from
The compound or a pharmaceutically acceptable salt thereof of claim 1, which is selected from

or a pharmaceutically acceptable salt thereof.
A pharmaceutical composition comprising a compound of any of claims 1-451 or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable excipient.
A method of inhibiting KRAS G12D activity in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of a composition of claim 52.
A method of treating a disease or disorder associated with KRAS G12D mutation in a subject in need thereof, comprising administering to the patient a therapeutically effective amount of a composition of claim 52.
The method of claim 54, wherein the disease or disorder associated with KRAS G12D mutation is cancer.
The method of claim 55, wherein the cancer is selected from carcinoma, squamous carcinoma, pancreatic cancer, prostate cancer, rectal cancer, colon cancer, colorectal cancer, non-small cell lung cancer, prostate cancer, small intestine cancer, sarcoma, leukemia, melanoma, and lymphoma.
The method of any of claims53-56, further comprising administering to the subject in need thereof an additional known anti-cancer agent.