CN113045565A

CN113045565A - Novel K-Ras G12C inhibitors

Info

Publication number: CN113045565A
Application number: CN201911386644.8A
Authority: CN
Inventors: 谢雨礼; 樊后兴; 曹刚; 钱立晖
Original assignee: Wigen Biomedicine Technology Shanghai Co Ltd
Current assignee: Wigen Biomedicine Technology Shanghai Co Ltd
Priority date: 2019-12-27
Filing date: 2019-12-27
Publication date: 2021-06-29
Also published as: WO2021129824A1; CN114901661A

Abstract

The invention relates to a compound shown in a formula (1) and a preparation method thereof, and application of the compound shown in the formula (1) and various optical isomers, various crystal forms and pharmaceutically acceptable salts thereof as an irreversible inhibitor of G12C mutant K-Ras protein in preparation of drugs for resisting Ras related diseases such as tumors and the like.

Description

Novel K-Ras G12C inhibitors

Technical Field

The invention belongs to the field of pharmaceutical chemistry, and particularly relates to a novel K-Ras G12C inhibitor, a preparation method thereof and a use method of the compound.

Background

Ras protein family is an important signal transduction transmission molecule in cells, and plays an important role in growth and development. Analysis and studies of a large number of in vitro tumor cells, animal models, and human tumor samples indicate that overactivation of Ras family proteins is an early event in human tumor development and is one of the important causes for the development and progression of a variety of cancers. Targeting and inhibition of Ras protein activity is therefore an important tool for the treatment of related tumors.

Ras proteins exist in two forms, which bind to GDP in an unactivated resting state; when cells receive signals such as growth factor stimulation, Ras proteins bind to GTP and are activated. Activated Ras proteins recruit a variety of signal-transfer proteins, promoting phosphorylation of downstream signaling molecules such as ERK, S6, thereby activating the Ras signaling pathway, regulating cell growth, survival, migration and differentiation. The Ras protein itself GTPase enzyme activity can be the hydrolysis of GTP back to GDP. The interaction of GTP enzyme activator proteins (GAPs) and Ras in cells greatly promotes the activity of Ras GTPase, so that the excessive activation of Ras proteins is prevented.

Mutations in the K-Ras, H-Ras and N-Ras proteins in the Ras protein family are one of the common genetic mutations in many tumors, and are the major factors that lead to the overactivation of Ras proteins in tumors. Compared with wild Ras protein, these mutations result in unregulated Ras protein activity, stable binding of GTP, and sustained activation, thereby promoting growth, migration, and differentiation of tumor cells. Among these, K-Ras protein mutations are the most common, accounting for 85% of all Ras mutations, while N-Ras (12%) and H-Ras (3%) are relatively rare. K-Ras mutations are very common in a variety of cancers: including pancreatic (95%), colorectal (45%), and lung (25%), among others, while relatively rare (< 2%) in breast, ovarian, and brain cancers. The K-Ras mutation site is mainly concentrated at position G12, with mutations at G12C being most common. For example, in non-small cell lung cancer (NSCLC), K-Ras G12C accounts for 50% of all K-Ras mutations, followed by G12V and G12D. Genomics research shows that K-Ras mutation in non-small cell lung cancer does not coexist with EGFR, ALK, ROS1, RET and BRAF mutation, but coexists with STK11, KEAP1, TP53 and other mutations, and suggests that the K-Ras mutation and the STK11, KEAP1, TP53 mutation and other synergistic effects are possibly involved in malignant transformation, proliferation and invasion of cells. In addition to tumors, abnormal activation of Ras protein is also involved in non-neoplastic diseases including diabetes, neurodegenerative diseases, and the like, and thus it can be seen that small molecule compounds targeting Ras protein can benefit a large number of cancer patients carrying specific genetic variations and non-cancer patients with overactivation of the Ras pathway.

Since forty years of discovery of Ras mutations in tumors, although we have made a more thorough understanding of the pathogenesis of the Ras pathway, no clinically effective therapeutic approach to targeting Ras proteins has been available for a large number of patients carrying Ras protein mutations and overactivation of the Ras pathway. Therefore, the development of a high-activity small-molecule inhibitor aiming at the Ras protein, in particular to the K-Ras G12C protein with high mutation frequency has important clinical significance.

Mirati reports in patents WO2017/201161, WO2019/099524 and US2019/270743 that K-Ras G12C inhibitors with tetrahydropyridopyrimidine as a parent nucleus have the disadvantages of poor or unstable activity, such as poor stability of compound A (Example 7) in US 2019/270743. In addition, some compounds have low activity, for Example, compound B (Example 494) in WO2019/099524 has antiproliferative activity on NCI-H358 cells of more than 1 μ M.

Therefore, there is a need to research and discover K-Ras G12C inhibitors with high activity and good stability.

Disclosure of Invention

The invention aims to provide a compound with a structural general formula shown in formula (1), or optical isomers, crystal forms, pharmaceutically acceptable salts, hydrates or solvates thereof:

in formula (1):

y is a bond or C1-C6 alkylene;

R¹is aryl or heteroaryl, which may be substituted with 1to 3 of the following groups: halogenA hydroxy group, an amino group, a C1-C3 alkyl group, a C2-C4 alkenyl group, a C3-C6 cycloalkyl group, a C1-C3 alkoxy group, a halogen-substituted C1-C3 alkyl group, or a halogen-substituted C1-C3 alkoxy group, and when substituted with a plurality of substituents, the substituents may be the same or different;

R²is aminoalkyl, cycloalkyl, heterocycloalkyl, aryl or heteroaryl, each of which may be substituted with 1to 3 of the following groups: H. halogen, CN, OH, C1-C3 alkyl, C1-C3 alkoxy, C3-C6 cycloalkyl, - (C1-C3 alkyl) -cyano, - (C1-C3 alkyl) - (C1-C3 alkoxy), - (C1-C3 alkyl) - (C3-C6 cycloalkyl), deuterated C1-C3 alkyl, halogen-substituted C1-C3 alkyl, or halogen-substituted C1-C3 alkoxy, which when substituted with multiple substituents, may be the same or different.

Q is

Wherein R is³And R⁴Independently H, D, halogen or C1-C3 alkyl;

is a 4-7 membered saturated heterocycloalkyl, partially saturated heterocycloalkyl or heteroaryl group having at least one N atom, which may be substituted with 1-3 of the following groups: H. d, halogen,

CN、NMe₂、NEt₂SMe, C1-C3 alkyl, C3-C6 cycloalkyl, -C (O) O- (C1-C3 alkyl), -C (O) NH- (C1-C3 alkyl), -C (O) NMe₂、-C(O)NEt₂- (C1-C3 alkyl) -NMe₂- (C1-C3 alkyl) -NEt₂Halogen-substituted C1-C3 alkyl or cyano-substituted C1-C3 alkyl, which when substituted with a plurality of substituents may be the same or different;

R⁵is C3-C6 cycloalkyl, - (C1-C3 alkyl) -cyano, - (C1-C3 alkyl) -hydroxy, - (C1-C3 alkyl) - (C3-C6 cycloalkyl), - (C1-C3 alkane- (C1-C3 alkoxy), - (C1-C3 alkyl) - (halogen-substituted C1-C3 alkoxy), heterocycloalkyl, partially saturated heterocycloalkyl, heteroaryl, - (C1-C3 alkyl) -heterocycloalkyl, - (C1-C3 alkyl) -heteroaryl or- (C1-C3 alkyl) -NR⁶R⁷The heterocycloalkyl, partially saturated heterocycloalkyl, heteroaryl, - (C1-C3 alkyl) -heterocycloalkyl or- (C1-C3 alkyl) -heteroaryl may be substituted with 1to 3 of the following groups: halogen, OH,

CN、NMe₂、NEt₂-C (O) O- (C1-C3 alkyl), -C (O) NH- (C1-C3 alkyl), -C (O) NMe₂、-C(O)NEt₂C1-C3 alkyl, C3-C6 cycloalkyl, halogen-substituted C1-C3 alkyl or cyano-substituted C1-C3 alkyl, which when substituted with multiple substituents may be the same or different, wherein R is⁶And R⁷Independently H, C1-C3 alkyl, C3-C6 cycloalkyl, or halogen substituted C1-C3 alkyl.

In another preferred embodiment, wherein in the general formula (1), Y is a bond, -CH₂-, -CH (Me) -or-CH₂CH₂-。

In another preferred embodiment, wherein in said general formula (1), R¹Comprises the following steps:

wherein R is^aAnd R^bIndependently H, halogen, hydroxyl, amino, C1-C3 alkyl, C2-C4 alkenyl, C3-C6 cycloalkyl, C1-C3 alkoxy, halogen substituted C1-C3 alkyl or halogen substituted C1-C3 alkoxy.

In another preferred embodiment, wherein in said general formula (1), R²Comprises the following steps:

wherein n is 1,2 or3，R^cAnd R^dIndependently H, halogen, CN, OH, C1-C3 alkyl, C3-C6 cycloalkyl, C1-C3 alkoxy, halogen substituted C1-C3 alkyl or halogen substituted C1-C3 alkoxy, R^eIs C1-C3 alkyl, C3-C6 cycloalkyl, deuterated C1-C3 alkyl, - (C1-C3 alkyl) -cyano, - (C1-C3 alkyl) - (C1-C3 alkoxy), - (C1-C3 alkyl) - (C3-C6 cycloalkyl), deuterated C1-C3 alkyl, halogen-substituted C1-C3 alkyl or halogen-substituted C1-C3 alkoxy, and when substituted by a plurality of substituents, the substituents may be the same or different.

In another preferred embodiment, wherein in said general formula (1), Q is

Wherein R is³And R⁴Independently is H or D;

is composed of

In another preferred embodiment, wherein in said general formula (1), Q is

Wherein R is⁵Is composed of

In various embodiments, representative compounds of the invention have one of the following structures:

another object of the present invention is to provide a pharmaceutical composition comprising a pharmaceutically acceptable excipient or carrier and, as an active ingredient, a compound of the general formula (1) of the present invention, or each of its optical isomers, a pharmaceutically acceptable inorganic or organic salt.

Still another object of the present invention is to provide the use of the above-mentioned compound of the present invention, or each optical isomer, pharmaceutically acceptable inorganic or organic salt thereof, for the preparation of a medicament for treating RAS-related diseases.

Through synthesis and careful study of various novel compounds with K-RAS G12C inhibiting effect, the inventors found that in the compounds of formula (1), when Q is enamine structure

In which

When the compound is heteroaryl or substituted heteroaryl, the K-RAS G12C inhibitory activity of the compound is greatly improved, and the compound has good stability, while the compound in which the aryl group is connected by a carbon-carbon single bond and acrylamide in the patent WO2019/099524 has poor activity, such as that the antiproliferative activity of the compound B on NCI-H358 cells is more than 1 mu M; in addition to this, the present invention is,

when the compound is heterocyclic alkyl or lacks electron-withdrawing groups such as CO beside an N atom, the stability of the compound is poor. On the other hand, the present inventors have found 2 (R) of acrylamide⁵Group) with a bulky or substituted heteroaryl group, the compounds also have good K-RAS G12C inhibitory activity and compound stability, whereas the 2-position of acrylamide in patent US2019/270743 with a smaller group has poor compound stability, such as compound a listed above.

It is to be understood that both the foregoing general description and the following detailed description of the present invention are exemplary and explanatory and are intended to provide further explanation of the invention as claimed.

Synthesis of Compounds

The following specifically describes the production process of the compound of the general formula (1) of the present invention, but these specific processes do not set any limit to the present invention.

The compounds of formula (1) described above may be synthesized using standard synthetic techniques or known techniques in combination with the methods described herein. In addition, the solvents, temperatures and other reaction conditions mentioned herein may vary. The starting materials for the synthesis of the compounds may be obtained synthetically or from commercial sources, e.g.,but are not limited to, Aldrich Chemical co. (Milwaukee, Wis.) or Sigma Chemical co. (st. The compounds described herein and other related compounds having various substituents can be synthesized using well-known techniques and starting materials, including those found in March, A_DVANCED O_RGANIC C_HEMISTRY4^thEd., (Wiley 1992); carey and Sundberg, A_DVANCED O_RGANIC C_HEMISTRY 4^thEd, Vols.A and B (Plenum2000, 2001), Green and Wuts, P_ROTECTIVE G_ROUPSIN O_RGANIC S_YNTHESIS 3^rdThe method in ed., (Wiley 1999). The general method of compound preparation may be varied by the use of appropriate reagents and conditions for introducing different groups into the formulae provided herein.

In one aspect, the compounds described herein are according to methods well known in the art. However, the conditions of the method, such as reactants, solvent, base, amount of the compound used, reaction temperature, time required for the reaction, and the like, are not limited to the following explanation. The compounds of the present invention may also be conveniently prepared by optionally combining various synthetic methods described in the present specification or known in the art, and such combinations may be readily carried out by those skilled in the art to which the present invention pertains. In one aspect, the present invention also provides a method for preparing the compound represented by the general formula (1), which is prepared by the following general reaction scheme 1, scheme 2 or scheme 3:

general reaction scheme 1

Embodiments of the compounds of formula (1) may be prepared according to general reaction scheme 1 (method A), wherein R¹、R²Y and Q are as defined above, PG and PG¹Represents different protecting groups of amine groups, and X represents Br, I, trifluoromethanesulfonate, boric acid, borate or potassium fluoroborate. As shown in general reaction scheme 1, the raw material A1 (synthesized in reference patent WO 2017/201161) undergoes oxidation reaction to obtain compound A2, compound A2 and compound R²-Y-OH under basic conditionsThe reaction is carried out to obtain a compound A3, the compound A3 removes the protecting group PG (e.g., Boc) to obtain a compound A4, a compound A4 and R¹Coupling with-X to give compound A5, Compound A5 deprotecting the protecting group PG¹(e.g., Cbz) to give compound A6, and condensation reaction of compound A6 with Q-COOH gave the title compound A7.

General reaction scheme 2

Embodiments of the compounds of formula (1) may be prepared according to general reaction scheme 2 (method B), wherein R¹、R²Y and R⁵W represents Br or I, K represents boric acid, boric acid ester or potassium fluoroborate, R is as defined above⁵In (2) directly bonded to K is a carbon atom. As shown in general reaction scheme 2, condensation reaction of intermediate A6 and compound B1 gives compound B2, compound B2 and R⁵The Suzuki coupling reaction of-K gives the end product B3.

General reaction scheme 3

Embodiments of the compounds of formula (1) may be prepared according to general reaction scheme 3 (method C), wherein R¹、R²Y and R⁵W represents Br or I, R, as defined hereinbefore⁵Directly attached to H in (b) is a nitrogen atom. Intermediate B2 and Compound R as shown in general reaction scheme 2⁵And carrying out Ullmann coupling reaction on the-H to obtain a final product C1.

Further forms of the compounds

"pharmaceutically acceptable" as used herein refers to a substance, such as a carrier or diluent, which does not diminish the biological activity or properties of the compound and which is relatively non-toxic, e.g., by being administered to an individual without causing unwanted biological effects or interacting in a deleterious manner with any of the components it contains.

The term "pharmaceutically acceptable salt" refers to a form of a compound that does not cause significant irritation to the organism to which it is administered and does not abrogate the biological activity and properties of the compound. In certain particular aspects, pharmaceutically acceptable salts are obtained by reacting a compound of formula (1) with an acid, such as an inorganic acid, e.g., hydrochloric acid, hydrobromic acid, hydrofluoric acid, sulfuric acid, phosphoric acid, nitric acid, phosphoric acid, etc., an organic acid, e.g., formic acid, acetic acid, propionic acid, oxalic acid, trifluoroacetic acid, malonic acid, succinic acid, fumaric acid, maleic acid, lactic acid, malic acid, tartaric acid, citric acid, picric acid, methanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid, etc., and an acidic amino acid, e.g., aspartic acid, glutamic acid, etc.

References to pharmaceutically acceptable salts are understood to include solvent addition forms or crystalline forms, especially solvates or polymorphs. Solvates contain either stoichiometric or non-stoichiometric amounts of solvent and are selectively formed during crystallization with pharmaceutically acceptable solvents such as water, ethanol, and the like. Hydrates are formed when the solvent is water, or alcoholates are formed when the solvent is ethanol. Solvates of the compounds of formula (1) are conveniently prepared or formed as described herein. For example, the hydrate of the compound of formula (1) is conveniently prepared by recrystallization from a mixed solvent of water/organic solvent, using an organic solvent including, but not limited to, tetrahydrofuran, acetone, ethanol or methanol. In addition, the compounds mentioned herein can exist in unsolvated as well as solvated forms. In general, the solvated forms are considered equivalent to unsolvated forms for purposes of the compounds and methods provided herein.

In other embodiments, the compound of formula (1) is prepared in different forms, including, but not limited to, amorphous, pulverized, and nano-sized forms. In addition, the compound of formula (1) includes crystalline forms, and may also be polymorphic forms. Polymorphs include different lattice arrangements of the same elemental composition of a compound. Polymorphs typically have different X-ray diffraction patterns, infrared spectra, melting points, densities, hardness, crystal forms, optical and electrical properties, stability and solubility. Different factors such as recrystallization solvent, crystallization rate and storage temperature may cause a single crystal form to dominate.

In another aspect, the compounds of formula (1) have one or more stereogenic centers and thus occur as racemates, racemic mixtures, single enantiomers, diastereomeric compounds and single diastereomers. Asymmetric centers that may be present depend on the nature of the various substituents on the molecule. Each such asymmetric center will independently produce two optical isomers and all possible optical isomers and diastereomeric mixtures and pure or partially pure compounds are included within the scope of the invention. The present invention is meant to include all such isomeric forms of these compounds.

Term(s) for

Unless otherwise defined, terms used in this application, including the specification and claims, are defined as follows. It must be noted that, in the specification and the appended claims, the singular forms "a," "an," and "the" include plural referents unless the context clearly dictates otherwise. Conventional methods of mass spectrometry, nuclear magnetism, HPLC, protein chemistry, biochemistry, recombinant DNA technology and pharmacology are used, if not otherwise stated. In this application, "or" and "means" and/or "are used unless otherwise stated.

"alkyl" refers to a saturated aliphatic hydrocarbon group, including straight and branched chain groups of 1to 6 carbon atoms. Lower alkyl groups having 1to 4 carbon atoms are preferred, such as methyl, ethyl, propyl, 2-propyl, n-butyl, isobutyl, tert-butyl. As used herein, "alkyl" includes unsubstituted and substituted alkyl groups, especially alkyl groups substituted with one or more halogens. Preferred alkyl groups are selected from CH₃、CH₃CH₂、CF₃、CHF₂、CF₃CH、ⁱPr、ⁿPr、ⁱBu、ⁿBu or^tBu。

"cycloalkyl" refers to a 3-to 6-membered all-carbon monocyclic aliphatic hydrocarbon group in which one or more rings may contain one or more double bonds, but none of the rings has a fully conjugated pi-electron system. For example, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexane, cyclohexadiene and the like.

"alkoxy" means a linkage to the rest of the molecule through an ether oxygen atomA partial alkyl group. Representative of alkoxy groups are alkoxy groups having 1to 6 carbon atoms, such as methoxy, ethoxy, propoxy, isopropoxy, butoxy, isobutoxy, sec-butoxy and tert-butoxy. As used herein, "alkoxy" includes unsubstituted and substituted alkoxy groups, especially alkoxy groups substituted with one or more halogens. Preferred alkoxy groups are selected from OCH₃、OCF₃、CHF₂O、CF₃CH₂O、^i-PrO、^n-PrO、^i-BuO、^n-BuO or^t-BuO。

"aryl" refers to a group having at least one aromatic ring structure, i.e., a carbocyclic aryl group having a conjugated pi-electron system, such as a benzene ring and a naphthalene ring.

"heteroaryl" refers to an aromatic group containing one or more heteroatoms (O, S or N), the heteroaryl group being monocyclic or polycyclic, e.g., a monocyclic heteroaryl ring fused to one or more carbocyclic aromatic groups or other monocyclic heterocyclic groups. Examples of heteroaryl groups include, but are not limited to, pyridyl, pyridazinyl, imidazolyl, pyrimidinyl, pyrazolyl, triazolyl, pyrazinyl, quinolinyl, isoquinolinyl, tetrazolyl, furyl, thienyl, isoxazolyl, thiazolyl, oxazolyl, isothiazolyl, pyrrolyl, indolyl, benzimidazolyl, benzofuryl, benzothiazolyl, benzothienyl, benzoxazolyl, benzopyridyl, and pyrrolopyrimidinyl.

"heterocycloalkyl" refers to a saturated or partially unsaturated ring system radical containing one or more heteroatoms (O, S or N) in which the nitrogen and sulfur atoms are optionally oxidized and the nitrogen atoms are optionally quaternized as ring atoms. Unless otherwise indicated, the ring system of "heterocycloalkyl" may be a monocyclic, bicyclic, spiro or polycyclic ring system. "heterocycloalkyl" can be attached to the rest of the molecule through more than one ring carbon or heteroatom. Examples of "heterocycloalkyl" include, but are not limited to, pyrrolidine, piperidine, N-methylpiperidine, tetrahydroimidazole, pyrazolidine, butyrolactam, valerolactam, imidazolidinone, hydantoin, dioxolane, phthalimide, piperidine, pyrimidine-2, 4(1H,3H) -dione, 1, 4-dioxane, morpholine, thiomorpholine-S-oxide, thiomorpholine-S, S-oxide, piperazine, pyran, pyridone, 3-pyrroline, thiopyran, pyrone, tetrahydrofuran, tetrahydrothiophene, quinuclidine, 2-azaspiro [3.3] heptane and the like.

"halo" (or halo) means fluoro, chloro, bromo, or iodo. The term "halogen substituted" (or "halo") as appearing before the radical name means that the radical is partially or fully halo, that is, substituted in any combination by F, Cl, Br or I, preferably by F or Cl.

The term "bond" or "single bond" refers to a chemical bond between two atoms or between two moieties when the atoms connected by the bond are considered part of a larger structure. In one aspect, when a group described herein is a bond, the absence of a reference group allows for the formation of a bond between the remaining defined groups.

The term "membered ring" includes any cyclic structure. The term "element" is intended to mean the number of backbone atoms constituting a ring. Thus, for example, cyclohexyl, pyridyl, pyranyl, thiopyranyl are six-membered rings and cyclopentyl, pyrrolyl, furanyl and thienyl are five-membered rings.

The term "fragment" refers to a specific part or functional group of a molecule. Chemical moieties are generally considered to be chemical entities contained in or attached to a molecule.

Specific pharmaceutical and medical terms

The term "acceptable", as used herein, means that a prescribed component or active ingredient does not unduly adversely affect the health of the general therapeutic target.

The terms "treat," "treatment process," or "therapy" as used herein include alleviating, inhibiting, or ameliorating a symptom or condition of a disease; inhibiting the generation of complications; ameliorating or preventing underlying metabolic syndrome; inhibiting the development of a disease or condition, such as controlling the development of a disease or condition; alleviating the disease or symptoms; regression of the disease or symptoms; alleviating a complication caused by the disease or symptom, or preventing or treating a symptom caused by the disease or symptom.

As used herein, a compound or pharmaceutical composition, when administered, can ameliorate a disease, symptom, or condition, particularly severity, delay onset, slow progression, or reduce duration of a condition. Whether fixed or temporary, continuous or intermittent, may be attributed to or associated with administration.

"active ingredient" means a compound represented by the general formula (1), and a pharmaceutically acceptable inorganic or organic salt of the compound of the general formula (1). The compounds of the present invention may contain one or more asymmetric centers and thus occur as racemates, racemic mixtures, single enantiomers, diastereomeric compounds and individual diastereomers. Asymmetric centers that may be present depend on the nature of the various substituents on the molecule. Each such asymmetric center will independently produce two optical isomers and all possible optical isomers and diastereomeric mixtures and pure or partially pure compounds are included within the scope of the invention. The present invention is meant to include all such isomeric forms of these compounds.

The terms "compound", "composition", "medicament" or "drug" are used interchangeably herein and refer to a compound or composition that, when administered to an individual (human or animal), is capable of inducing a desired pharmaceutical and/or physiological response through local and/or systemic action.

The term "administering" or "administration" as used herein refers to the direct administration of the compound or composition, or the administration of a prodrug (produg), derivative (derivative), or analog (analog) of the active compound, and the like.

Although numerical ranges and parameters setting forth the broad scope of the invention are approximate, the values set forth in the specific examples are presented as precisely as possible. Any numerical value, however, inherently contains certain standard deviations found in their respective testing measurements. As used herein, "about" generally means that the actual value is within plus or minus 10%, 5%, 1%, or 0.5% of a particular value or range. Alternatively, the term "about" means that the actual value falls within the acceptable standard error of the mean, as considered by those skilled in the art. Except in the experimental examples, or where otherwise expressly indicated, it is to be understood that all ranges, amounts, values and percentages herein used (e.g., to describe amounts of materials, length of time, temperature, operating conditions, quantitative ratios, and the like) are to be modified by the word "about". Accordingly, unless indicated to the contrary, the numerical parameters set forth in the specification and attached claims are approximations that may vary depending upon the desired properties sought to be obtained. At the very least, these numerical parameters are to be understood as meaning both the number of significant digits indicated and the number resulting from applying ordinary rounding techniques.

Unless defined otherwise herein, the scientific and technical terms used herein have the same meaning as commonly understood by one of ordinary skill in the art. Furthermore, as used herein, the singular tense of a noun, unless otherwise conflicting with context, encompasses the plural form of that noun; the use of plural nouns also covers the singular form of such nouns.

Therapeutic uses

The invention provides methods of treating diseases, including but not limited to conditions (e.g., cancer) involving G12C K-Ras, G12C H-Ras and/or G12C N-Ras mutations, using the compounds or pharmaceutical compositions of the invention.

In some embodiments, there is provided a method for the treatment of cancer, the method comprising administering to an individual in need thereof an effective amount of a pharmaceutical composition of any of the foregoing compounds of protective structure (1). In some embodiments, the cancer is mediated by K-Ras, H-Ras and/or G12C N-Ras mutations. In other embodiments, the cancer is lung cancer, pancreatic cancer, colon cancer, MYH-related polyposis, or colorectal cancer.

Route of administration

The compound and the pharmaceutically acceptable salt thereof can be prepared into various preparations, wherein the preparation comprises the compound or the pharmaceutically acceptable salt thereof in a safe and effective amount range and a pharmaceutically acceptable excipient or carrier. Wherein "safe, effective amount" means: the amount of the compound is sufficient to significantly improve the condition without causing serious side effects. The safe and effective amount of the compound is determined according to the age, condition, course of treatment and other specific conditions of a treated subject.

"pharmaceutically acceptable excipient or carrier" refers to: one or more compatible solid or liquid fillers or gel substances which are suitable for human use and must be of sufficient purity and sufficiently low toxicity. By "compatible" is meant herein that the components of the composition are capable of being blended with the compounds of the present invention and with each other without significantly diminishing the efficacy of the compounds. Examples of pharmaceutically acceptable excipients or carrier moieties are cellulose and its derivatives (e.g. sodium carboxymethylcellulose, sodium ethylcellulose, cellulose acetate etc.), gelatin, talc, solid lubricants (e.g. stearic acid, magnesium stearate), calcium sulfate, vegetable oils (e.g. soybean oil, sesame oil, peanut oil, olive oil etc.), polyols (e.g. propylene glycol, glycerol, mannitol, sorbitol etc.), emulsifiers (e.g. propylene glycol, glycerol, mannitol, sorbitol etc.)

) Wetting agents (e.g., sodium lauryl sulfate), coloring agents, flavoring agents, stabilizers, antioxidants, preservatives, pyrogen-free water, and the like.

When the compounds of the present invention are administered, they may be administered orally, rectally, parenterally (intravenously, intramuscularly or subcutaneously), topically.

Solid dosage forms for oral administration include capsules, tablets, pills, powders and granules. In these solid dosage forms, the active compound is mixed with at least one conventional inert excipient (or carrier), such as sodium citrate or dicalcium phosphate, or with the following ingredients: (a) fillers or extenders, for example, starch, lactose, sucrose, glucose, mannitol and silicic acid; (b) binders, for example, hydroxymethylcellulose, alginates, gelatin, polyvinylpyrrolidone, sucrose and acacia; (c) humectants, for example, glycerol; (d) disintegrating agents, for example, agar-agar, calcium carbonate, potato or tapioca starch, alginic acid, certain complex silicates, and sodium carbonate; (e) slow solvents, such as paraffin; (f) absorption accelerators, e.g., quaternary ammonium compounds; (g) wetting agents, such as cetyl alcohol and glycerol monostearate; (h) adsorbents, for example, kaolin; and (i) lubricants, for example, talc, calcium stearate, magnesium stearate, solid polyethylene glycols, sodium lauryl sulfate, or mixtures thereof. In capsules, tablets and pills, the dosage forms may also comprise buffering agents.

Solid dosage forms such as tablets, dragees, capsules, pills, and granules can be prepared using coatings and shells such as enteric coatings and other materials well known in the art. They may contain opacifying agents and the release of the active compound or compounds in such compositions may be delayed in release in a certain part of the digestive tract. Examples of embedding components which can be used are polymeric substances and wax-like substances. If desired, the active compound may also be in microencapsulated form with one or more of the above-mentioned excipients.

Liquid dosage forms for oral administration include pharmaceutically acceptable emulsions, solutions, suspensions, syrups or tinctures. In addition to the active compounds, the liquid dosage forms may contain inert diluents commonly employed in the art such as, for example, water or other solvents, solubilizing agents and emulsifiers such as ethyl alcohol, isopropyl alcohol, ethyl carbonate, ethyl acetate, propylene glycol, 1, 3-butylene glycol, dimethylformamide and oils, in particular, cottonseed, groundnut, corn germ, olive, castor and sesame oils or mixtures of such materials and the like.

In addition to these inert diluents, the compositions can also contain adjuvants such as wetting agents, emulsifying and suspending agents, sweetening, flavoring, and perfuming agents.

Suspensions, in addition to the active compounds, may contain suspending agents as, for example, ethoxylated isostearyl alcohols, polyoxyethylene sorbitol and sorbitan esters, microcrystalline cellulose, aluminum methoxide and agar, or mixtures of these substances, and the like.

Compositions for parenteral injection may comprise physiologically acceptable sterile aqueous or anhydrous solutions, dispersions, suspensions or emulsions, and sterile powders for reconstitution into sterile injectable solutions or dispersions. Suitable aqueous and nonaqueous carriers, diluents, solvents or vehicles include water, ethanol, polyols and suitable mixtures thereof.

Dosage forms for topical administration of the compounds of the present invention include ointments, powders, patches, sprays, and inhalants. The active ingredient is mixed under sterile conditions with a physiologically acceptable carrier and any preservatives, buffers, or propellants which may be required if necessary.

The compounds of the present invention may be administered alone or in combination with other pharmaceutically acceptable compounds. When the pharmaceutical composition is used, a safe and effective amount of the compound of the present invention is suitable for mammals (such as human beings) to be treated, wherein the administration dose is a pharmaceutically-considered effective administration dose, and for a human body with a weight of 60kg, the daily administration dose is usually 1to 2000mg, preferably 50 to 100 mg. Of course, the particular dosage will depend upon such factors as the route of administration, the health of the patient, and the like, and is within the skill of the skilled practitioner.

The features mentioned above with reference to the invention, or the features mentioned with reference to the embodiments, can be combined arbitrarily. All the features disclosed in this specification may be combined in any combination, and each feature disclosed in this specification may be replaced by alternative features serving the same, equivalent or similar purpose. Thus, unless expressly stated otherwise, the features disclosed are merely generic examples of equivalent or similar features.

The various specific aspects, features and advantages of the compounds, methods and pharmaceutical compositions described above are set forth in detail in the following description, which makes the present invention clear. It should be understood herein that the detailed description and examples, while indicating specific embodiments, are given by way of illustration only. After reading the description of the invention, one skilled in the art can make various changes or modifications to the invention, and such equivalents fall within the scope of the invention as defined in the application.

In all of the embodiments described herein, the first,¹H-NMR was recorded using a Vian Mercury 400 NMR spectrometer with chemical shifts expressed in delta (ppm); the silica gel used for separation is not illustrated to be 200-300 meshes, and the proportions of the eluents are volume ratios.

The invention employs the following abbreviations: ar represents argon; BFMO represents N1, N2-bis (furan-2-ylmethyl) oxyalkanamide; CDCl₃Represents deuterated chloroform; CH (CH)₃CN represents acetonitrile; cs₂CO₃Represents cesium carbonate; CuI represents cuprous iodide; DCM represents dichloromethane; DIPEA stands for diisopropylethylamine; dioxane represents 1, 4-Dioxane; DMF represents dimethylformamide; DMSO represents divaricate knotweed herb; EA represents ethyl acetate; EtOH stands for ethanol; h represents hour; NaOH represents sodium hydroxide; KOAc represents potassium acetate; LC-MS stands for liquid-mass spectrometry; m-CPBA represents m-chloroperoxybenzoic acid; MeOH represents methanol; min represents min; MS represents mass spectrum; na (Na)₂SO₄Represents sodium sulfate; NMR stands for nuclear magnetic resonance; pd₂(dba)₃Represents tris (dibenzylideneacetone) dipalladium; pd (dppf) Cl₂Represents 1,1' -bis (diphenylphosphino) ferrocene]Palladium dichloride; PE represents petroleum ether; ruphos stands for 2-bicyclohexylphosphine-2 ',6' -diisopropyloxybiphenyl;^t-BuOK for potassium tert-butoxide; T3P represents 1-propylphosphoric anhydride; TFA (CF)₃COOH) represents trifluoroacetic acid; THF represents tetrahydrofuran; toluene stands for Toluene.

The specific implementation mode is as follows:

preparation example Synthesis of- ((S) -4- (7- (8-chloronaphthalen-1-yl) -2- (((S) -1-methylpyrrolidin-2-yl) methoxy) -5,6,7, 8-tetrahydropyrido [3,4-d ] pyrimidin-4-yl) piperazin-2-yl) acetonitrile (A-6a)

(S) -4- (4- ((benzyloxy) carbonyl) -3- (cyanomethyl) piperazin-1-yl) -2- (methylsulfonyl) -5, 8-dihydropyrido [3,4-d ] pyrimidine-7 (6H) -carboxylic acid tert-butyl ester (A-2)

A-1(72.5g,134.6mmol) and DCM (725mL) were added to a 1L single-neck flask, cooled to 0-5 ℃ in an ice-salt bath under the protection of Ar, and m-CPBA (81.3g,471.1mmol) was added and the reaction was continued for 2 hours. After the completion of the reaction was monitored by LC-MS, the mixture was filtered, the filter cake was washed with DCM (50 mL. times.3), a saturated sodium bicarbonate solution (200mL) and a saturated sodium thiosulfate solution (300mL) were added to the filtrate, stirred at room temperature for 30min, the organic phase was separated, washed successively with a saturated sodium thiosulfate solution (200 mL. times.2) and a saturated sodium chloride solution (200mL), anhydrous Na₂SO₄Drying, filtering, and concentrating the filtrate to obtain crude extractProduct A-2(80g, 100% yield), ESI-MS M/z 571.3[ M + H ]]⁺。

4- ((S) -4- ((benzyloxy) carbonyl) -3- (cyanomethyl) piperazin-1-yl) -2- (((S) -1-methylpyrrolidin-2-yl) methoxy) -5, 8-dihydropyrido [3,4-d ] pyrimidine-7 (6H) -carboxylic acid tert-butyl ester (A-3a)

A-2(80g,134.6mmol), THF (600mL) and (S) - (1-methylpyrrolidin-2-yl) methanol (31g,269.2mmol) were added to a 1L single-neck flask, cooled to 0-5 ℃ in an ice-salt bath under Ar protection, then sodium tert-butoxide (32.3g,336.5mmol) was added and the reaction was continued for 2 h. After TLC and LC-MS to monitor the reaction was complete, the system was quenched with EA (300mL), saturated ammonium chloride solution (200mL), stirred, separated, the aqueous phase was extracted with EA (200mL), the organic phases were combined and washed with saturated sodium chloride solution (200 mL. times.2), concentrated, and the residue was purified by column chromatography (DCM/MeOH. 50/1to 30/1) to give product A-3a as a yellowish brown oil (60.5g, 74% yield), ESI-MS M/z:606.3[ M + H/M ])]⁺。

(S) -benzyl 2- (cyanomethyl) -4- (2- (((S) -1-methylpyrrolidin-2-yl) methoxy) -5,6,7, 8-tetrahydropyrido [3,4-d ] pyrimidin-4-yl) piperazine-1-carboxylate (A-4a)

Adding A-3a (60.5g,99.88mmol) and DCM (150mL) into a 1L single-neck bottle, cooling the mixed solution to 0-5 ℃ by using an ice salt bath under the protection of Ar, then adding TFA (228g,2mol), stirring and reacting for 2h at r.t., monitoring the reaction by LC-MS, concentrating the system, adding DCM (300mL) into the residue, adjusting the pH to 8-9 by using a saturated sodium carbonate solution, stirring, separating liquid, extracting the aqueous phase by using DCM (200mL 5), combining the organic phases, and using anhydrous Na for the organic phase₂SO₄Drying, filtering, concentrating the filtrate to obtain yellow foamy solid product A-4a (54.2g, yield 107%), ESI-MS M/z 506.3[ M + H ]]⁺。

(S) -benzyl 4- (7- (8-chloronaphthalen-1-yl) -2- (((S) -1-methylpyrrolidin-2-yl) methoxy) -5,6,7, 8-tetrahydropyrido [3,4-d ] pyrimidin-4-yl) -2- (cyanomethyl) piperazine-1-carboxylate (A-5a)

A1L single-neck bottle is charged with A-4a (45.5g,90.1mmol), 1-bromo-8-chloronaphthalene (32.6g,135mmol), cesium carbonate (73.3g,225mmol), Ruphos (8.4g,18mmol) and toluene (800mL), and after replacement protection of the mixed solution with Ar, Pd is added₂(dba)₃(16.5g,18mmol), heating to 90 ℃ under the protection of Ar, and stirring for reactionAnd 6 h. After the completion of the reaction was monitored by LC-MS, the system was filtered while hot, the filter cake was washed with DCM (100mL), the filtrate was concentrated and purified by column chromatography (DCM/MeOH 50/1to 20/1) to give the product A-5a as a yellowish brown foamy solid (36.6g, yield: 61%), ESI-MS M/z:666.4[ M + H ]: 666.4]⁺。

2- ((S) -4- (7- (8-chloronaphthalen-1-yl) -2- (((S) -1-methylpyrrolidin-2-yl) methoxy) -5,6,7, 8-tetrahydropyrido [3,4-d ] pyrimidin-4-yl) piperazin-2-yl) acetonitrile (A-6a)

A-5a (40.9g,61.39mmol), THF (100mL), MeOH (600mL), and 10% Pd/C (5g, wet% ═ 50%), H, were added to a 1L single-neck flask₂After three times of replacement, the mixture is stirred vigorously at room temperature and normal pressure for reaction for 20 hours. After the LC-MS monitoring reaction is completed, the system is added with ammonia methanol solution (20mL,7M), stirred for 15min and then filtered by diatomite, the filtrate is concentrated to dryness to obtain a yellow-brown foamy solid product A-6a (31.3g, yield: 95%), ESI-MS M/z:532.3[ M + H ], (yield: 95%) (]⁺。

Preparation examples 2-10 Synthesis of intermediates A-6b to A-6j

Different raw materials are utilized, and a synthesis method of the intermediate A-6a is adopted to obtain the target intermediates A-6b to A-6 j.

TABLE 1

2- ((S) -1- (2-bromoacryloyl) -4- (7- (8-chloronaphthalen-1-yl) -2- (((S) -1-methylpyrrolidin-2-yl) methoxy) -5,6,7, 8-tetrahydropyrido [3,4-d ] pyrimidin-4-yl) piperazin-2-yl) acetonitrile (B-2a)

A-6a (10.64g,20.0mmol) and 2-bromoacrylic acid (B-1, 9.06g,60mmol), DIPEA (25.8g,200mmol) and DMF (200mL) are added into a 100mL single-neck bottle, and T is slowly dropped at room temperature under the protection of mixed liquid Ar₃P (50.9g, 50% in DMF,80mmol) was added dropwise, the mixture was stirred at room temperature for 1H and quenched with water (200mL), EA (200 mL. times.2) was extracted, the organic phases were combined, concentrated, and the residue was purified by column chromatography (DCM/MeOH 50/1to 20/1 to give a light brown solid (5.45g, 41% yield), ESI-MS M/z 664.2/666.2[ M + H ] M + H]⁺。

Preparation examples 12-20 Synthesis of intermediates B-2B to B-2j

Different raw materials are utilized, and the target intermediates B-2B to B-2j are obtained by adopting a synthesis method of the intermediate B-2 a.

TABLE 2

Example Synthesis of- ((S) -1- ((E) -3- (1H-1,2, 4-triazol-1-yl) acryloyl) -4- (7- (8-chloronaphthalen-1-yl) -2- (((S) -1-methylpyrrolidin-2-yl) methoxy) -5,6,7, 8-tetrahydropyrido [3,4-d ] pyrimidin-4-yl) piperazin-2-yl) acetonitrile (Compound 1)

Compound 1 was prepared according to method a as described below:

2- ((S) -1- ((E) -3- (1H-1,2, 4-triazol-1-yl) acryloyl) -4- (7- (8-chloronaphthalen-1-yl) -2- (((S) -1-methylpyrrolidin-2-yl) methoxy) -5,6,7, 8-tetrahydropyrido [3,4-d ] pyrimidin-4-yl) piperazin-2-yl) acetonitrile

A100 mL single-neck flask was charged with A-6a (369mg,0.693mmol), (E) -3- (1H-1,2, 4-triazol-1-yl) acrylic acid (145mg,1.04mmol), DIPEA (269mg,2.08mmol) and DMF (10mL), and the mixture was added dropwise with T under Ar at room temperature₃P (662mg, 50% in DMF,1.04mmol), stirred at RT for 1h and quenched with water, EA (10 mL. times.2) extracted, the organic phases combined, concentrated and the residue pre-TLC (DCM/MeOH/NH)₃.H₂O-500/20/1) to give compound 1(205mg, 45% yield) as a pale yellow solid.

¹H NMR(400MHz,CDCl₃)δ:8.32(s,1H),8.08(d,J＝14.0Hz,2H),7.73(d,J＝8.1Hz,1H),7.60(t,J＝7.2Hz,1H),7.50(d,J＝7.5Hz,1H),7.41(dd,J＝11.5,7.9Hz,1H),7.31(t,J＝7.8Hz,1H),7.24-7.16(m,1H),7.12(d,J＝9.6Hz,1H),5.14(s,1H),4.52-4.34(m,2H),4.15(dt,J＝35.4,22.5Hz,4H),3.90-3.76(m,1H),3.57(s,1H),3.45(d,J＝13.9Hz,1H),3.13(td,J＝42.7,39.9,21.0Hz,5H),2.86-2.69(m,2H),2.55(dd,J＝28.8,9.7Hz,4H),2.33(s,1H),2.06(s,1H),1.77(s,4H)；ESI-MS m/z:653.4[M+H]⁺.

Example 2-example 85 Synthesis of Compound 2-Compound 85

Compound 2-compound 85 can be obtained by using intermediates a-6a to a-6j as starting materials and using synthesis method a, similar to the synthesis method of compound 1.

TABLE 3

Example Synthesis of 862- ((S) -4- (7- (8-chloronaphthalen-1-yl) -2- (((S) -1-methylpyrrolidin-2-yl) methoxy) -5,6,7, 8-tetrahydropyrido [3,4-d ] pyrimidin-4-yl) -1- (2-phenylpropenoyl) piperazin-2-yl) acetonitrile (Compound 86)

Compound 86 was prepared according to method B as described below:

2- ((S) -4- (7- (8-chloronaphthalen-1-yl) -2- (((S) -1-methylpyrrolidin-2-yl) methoxy) -5,6,7, 8-tetrahydropyrido [3,4-d ] pyrimidin-4-yl) -1- (2-phenylpropenoyl) piperazin-2-yl) acetonitrile

A50 mL single neck flask was charged with B-2a (50mg,0.075mmol), phenylboronic acid (18mg,0.15mmol), KOAc (19mg,0.192mmol), Dioxane (5mL) and H₂O (1mL), Ar replacement protected, Pd (dppf) Cl₂(10mg), rapidly heating the mixed solution to reflux under the protection of Ar, stirring for reacting for 20min, adding water (5mL) and EA (10mL) into the system, stirring, separating the solution,the organic phase was concentrated and the residue was pre-TLC (DCM/MeOH/NH)₃.H₂O-500/20/1) to give the product (8mg, 16% yield).

¹H NMR(400MHz,CDCl₃)δ:7.75(dt,J＝8.1,1.6Hz,1H),7.61-7.52(m,1H),7.45-7.31(m,7H),7.22(dt,J＝7.5,1.5Hz,1H),7.17-7.02(m,1H),6.03(s,1H),5.82(s,1H),5.04(m,1H),4.44-4.24(m,2H),4.15-4.05(m,2H),4.00-3.71(m,3H),3.52(d,J＝11.3Hz,2H),3.43-3.33(m,1H),3.26-2.76(m,7H),2.63(q,J＝6.5,5.9Hz,1H),2.54(d,J＝14.1Hz,1H),2.42(s,3H),2.22(dt,J＝9.8,7.3Hz,1H),2.03(d,J＝11.9Hz,1H),1.89-1.75(m,2H)；ESI-MS m/z:663.2[M+H]⁺.

EXAMPLE 87 EXAMPLE 127 Compound 87 Synthesis of Compound 127

Using intermediates B-2a to B-2j as starting materials, compound 87-compound 127 can be obtained by synthesis method B, which is similar to the synthesis method of compound 86.

TABLE 4

Example Synthesis of 1282- ((S) -1- (2- (1H-imidazol-1-yl) acryloyl) -4- (7- (8-chloronaphthalen-1-yl) -2- (((S) -1-methylpyrrolidin-2-yl) methoxy) -5,6,7, 8-tetrahydropyrido [3,4-d ] pyrimidin-4-yl) piperazin-2-yl) acetonitrile (Compound 128)

Compound 128 was prepared according to procedure C as described below:

2- ((S) -1- (2- (1H-imidazol-1-yl) acryloyl) -4- (7- (8-chloronaphthalen-1-yl) -2- (((S) -1-methylpyrrolidin-2-yl) methoxy) -5,6,7, 8-tetrahydropyrido [3,4-d ] pyrimidin-4-yl) piperazin-2-yl) acetonitrile

A10 mL single-necked flask was charged with B-2a (25mg,0.038mmol), imidazole (4mg,0.056mmol), CuI (2mg,0.008mmol), Cs₂CO₃(33mg,0.098mmol), DMSO (2mL) and BFMO (2mg,0.008mmol), the mixture was heated to 80 ℃ under Ar protection and stirred for 1 h. After MS monitoring that the basic reaction of the raw materials is complete, EA (10mL) and H are added to the mixture₂O (10mL), stirred, separated, the organic phase concentrated and the residue subjected to pre-TLC (DCM/MeOH/NH)₃.H₂O-500/20/1) to give compound 128(5mg, 20% yield).

¹H NMR(400MHz,DMSO-d₆)δ:8.22(s,1H),8.10-8.03(m,1H),7.93(d,J＝7.6Hz,1H),7.87(s,1H),7.78-7.72(m,1H),7.57(dd,J＝18.4,7.7Hz,2H),7.49-7.42(m,1H),7.38(s,2H),7.10(s,1H),5.33(t,J＝4.7Hz,1H),5.03(s,2H),4.66(s,1H),4.42(s,1H),4.34-4.12(m,4H),4.06(s,3H),3.22-3.00(m,5H),2.94(s,3H),2.69-2.66(m,1H),2.37-2.32(m,3H),2.05-1.90(m,3H)；ESI-MS m/z:652.3[M+H]⁺.

EXAMPLE 129 Synthesis of example 148 Compound 129 Compound 148

Compound 129-compound 148 can be obtained using intermediates B-2a to B-2j as starting materials, using synthesis method C, analogous to the synthesis method of compound 128.

TABLE 5

EXAMPLE 149 detection of the amount of pERK and ERK proteins in H358 cells by Compounds

H358 cells were seeded in a 24-well plate, after one day of growth, a test compound (1. mu.M concentration) was added, after 24 hours of action of the compound, after lysis of the cells, cell lysates were transferred to a 96-well ELISA plate, levels of pERK and ERK in the lysates were determined using an ELISA kit (abcam 176660), the ratio of pERK to ERK was calculated and compared to DMSO group, the percentage of inhibition of pERK activity by the compound was calculated, and the results are shown in Table 6 below.

TABLE 6 inhibitory Activity of Compounds of the invention against pERK levels in H358 cells

+ represents an inhibition rate of less than or equal to 50%

+ indicates an inhibition of 50% to 90%

And +++ indicates an inhibition of greater than 90%.

EXAMPLE 150 antiproliferative Activity of Compounds on H358 cells

2500H 358 cells were plated in ultra low adsorption 96-well plates (corning,7007) and after one day of growth, a gradient dilution compound (5. mu.M maximum, 5-fold dilution, five total doses) was added, and after three days of compound addition, Cell Titer Glow (Promega, G9681) was added to evaluate pellet growth and calculate IC₅₀The values, results are given in table 7 below.

TABLE 7 antiproliferative activity of the compounds of the invention on H358 cells

+ represents a compound of IC₅₀Greater than 1 μ M

+ represents the IC of the compound₅₀Is 0.3 to 1. mu.M

+ + + + + denotes IC of the Compound₅₀Less than 0.3. mu.M.

As is apparent from the activity data in tables 7 and 8 above, in the compound of the general formula (1), when Q is an enamine structure

In which

In the case of heteroaryl or substituted heteroaryl, the K-RAS G12C inhibitory activity of the compound is greatly improved, while the compound in which the heteroaryl is connected with acrylamide through a carbon-carbon single bond in the patent WO2019/099524 has poor activity, such as the antiproliferative activity of the compound B on NCI-H358 cells is more than 1 mu M. The carbon-nitrogen single bond is presumed to have stronger dipole moment than the carbon-carbon single bond, and the nitrogen atom loses partial electrons, which is favorable for stabilizing the intermediate state of the cysteine of the K-RAS protein after the acrylamide addition, so that the compound has strong activity. On the other hand, when the compound of the present invention has a bulky heteroaryl or substituted heteroaryl substituent at the 2-position of acrylamide, such as imidazolyl or pyrimidinyl, the compound also has excellent K-RAS G12C inhibitory activity, presumably because the N atom on the heteroaryl group facilitates binding to K-RAS protein.

Stability testing of the Compounds of example 151

Accurately weighing two parts (2-3 mg each) of the test compound, storing one part at the temperature of-20 ℃ in a refrigerator, placing one part in a room temperature, dissolving the sample in a 50mL volumetric flask after 72 hours, adding a proper amount of methanol, adding water to a constant volume to reach a scale, and shaking up to obtain the solution to be tested. The HPLC was used to measure the area of the peak of the liquid phase for both solutions and to determine the area of the peak of the liquid phase per unit concentration of the sample placed at room temperature divided by the area of the peak of the unit concentration of the sample stored at-20 ℃ in the refrigerator, the percentage obtained representing the residual amount of the compound after 72 hours of room temperature placement.

TABLE 8 stability of the Compounds after 72 hours at Room temperature

As shown by the stability data of Compound A in Table 9, the poor stability of the compound is presumed to be caused by intermolecular polymerization of the basic nitrogen atom in the molecule and the double bond of acrylamide. Compared with the contrast medicament A, the compound of the invention carries out the substitution of a group with larger volume on acrylamide, so that the stability of the compound of the invention is greatly improved, which has very important significance for the process synthesis, quality control and pharmacy of the compound.

Example 152 evaluation of antitumor Activity in mice

Human pancreatic cancer Mia PaCa-2 cells were treated with 1640 containing 10% fetal bovine serum at 37 ℃ with 5% CO₂And (4) performing conventional culture in an incubator, and collecting cells after passage until the cells reach the required amount. Will be 1 × 10⁷A Mia PaCa-2 cell was injected into the left dorsal side of each nude mouse, and the tumor was allowed to grow to 150mm³Thereafter, animals were randomized into groups to begin dosing. 1) solvent control group, 8 respectively; 2) compound 1, compound 5, compound 70, compound 92, compound 97 and compound 121 groups, 8 per group. The solvent control group was intragastrically administered with 0.5% CMC-Na once a day; compound 1, compound 5, compound 70, compound 92, compound 97 and compound 121 groups were gavaged once daily with a 0.5% CMC-Na suspension. Tumor volumes were measured two and four weeks per week, mouse body weights were measured, and nude mice were sacrificed on day 21 of dosing, and the test results are shown in table 9 below.

TABLE 9 Experimental therapeutic Effect of Compounds on human pancreatic cancer Mia PaCa-2 nude mouse transplantable tumors

Compound (I)	Dosage (mg/kg)	Dosing regimens	Anti-tumor effect
				1	10	qd*21	67% inhibition
5	10	qd*21	63% inhibition
				70	10	qd*21	77% inhibition
92	10	qd*21	72% inhibition
				121	10	qd*21	Inhibition of 76%
A	10	qd*21	58% inhibition

Compared with a contrast medicament A, the compound has stronger in-vitro activity and compound stability, and meanwhile, an in-vivo anti-tumor test also shows that the compound has better in-vivo activity, so that the compound has better drug effect and drug forming property in clinical application.

Claims

1. A compound with a structure shown as a general formula (1) or each optical isomer, each crystal form, pharmaceutically acceptable salt, hydrate or solvate thereof:

in formula (1):

y is a bond or C1-C6 alkylene;

R¹is aryl or heteroaryl, which may be substituted with 1to 3 of the following groups: halogen, hydroxy, amino, C1-C3 alkyl, C2-C4 alkenyl, C3-C6 cycloalkyl, C1-C3 alkoxy, halogen-substituted C1-C3 alkyl or halogen-substituted C1-C3 alkoxy, and when substituted with a plurality of substituents, the substituents may be the same or different;

Q is

Wherein R is³And R⁴Independently H, D, halogen or C1-C3 alkyl;

R⁵is C3-C6 cycloalkyl, - (C1-C3 alkyl) -cyano, - (C1-C3 alkyl) -hydroxy, - (C1-C3 alkyl) - (C3-C6 cycloalkyl), - (C1-C3 alkyl) - (C1-C3 alkoxy), - (C1-C3 alkyl) - (halogen substituted C1-C3 alkoxy), heterocycloalkyl, partially saturated heterocycloalkyl, heteroaryl, - (C1-C3 alkyl) -heterocycloalkyl, - (C1-C3 alkyl) -heteroaryl or- (C1-C3 alkyl) -NR⁶R⁷The heterocycloalkyl, partially saturated heterocycloalkyl, heteroaryl, - (C1-C3 alkyl) -heterocycloalkyl or- (C1-C3 alkyl) -heteroaryl may be substituted with 1to 3 of the following groups: halogen, OH,

2. The compound according to claim 1, wherein in the general formula (1), Y is a bond, -CH₂-, -CH (Me) -or-CH₂CH₂-。

3. The compound according to claim 1-2, wherein in the general formula (1), R is¹Comprises the following steps:

4. A compound according to claims 1to 3, wherein in the general formula (1), R is²Comprises the following steps:

wherein n is 1,2 or 3, R^cAnd R^dIndependently H, halogen, CN, OH, C1-C3 alkyl, C3-C6 cycloalkyl, C1-C3 alkoxy, halogen substituted C1-C3 alkyl or halogen substituted C1-C3 alkoxy, R^eIs C1-C3 alkyl, C3-C6 cycloalkyl, deuterated C1-C3 alkyl, - (C1-C3 alkyl) -cyano, - (C1-C3 alkyl) - (C1-C3 alkoxy), - (C1-C3 alkyl) - (C3-C6 cycloalkyl), deuterated C1-C3 alkyl, halogen-substituted C1-C3 alkyl or halogen-substituted C1-C3 alkoxy, and when substituted by a plurality of substituents, the substituents may be the same or different.

5. As claimed in claims 1-4Wherein in the general formula (1), Q is

Wherein R is³And R⁴Independently is H or D;

is composed of

6. The compound according to claim 1to 4, wherein in the general formula (1), Q is

Wherein R is⁵Is composed of

7. The compound of claims 1-6, or a pharmaceutically acceptable salt thereof, wherein the compound has one of the following structures:

8. a pharmaceutical composition for the treatment, regulation and/or prevention of diseases associated with the K-Ras G12C mutant protein, which comprises a pharmaceutically acceptable excipient or carrier, and as an active ingredient, a compound according to any one of claims 1to 7, or each optical isomer, pharmaceutically acceptable salt, hydrate or solvate thereof.

9. Use of a compound according to any one of claims 1to 7, or each optical isomer, each crystalline form, a pharmaceutically acceptable salt, hydrate or solvate thereof, for the treatment of a disorder mediated by the K-Ras G12C mutation in a subject in need thereof.

10. The method of claim 9 wherein the condition is cancer, and the cancer is hematological cancer and solid tumors.