CN115716840A - Heterocyclic compound with KRAS (KRAS-mutated protein) inhibition effect and pharmaceutical composition and application thereof - Google Patents

Heterocyclic compound with KRAS (KRAS-mutated protein) inhibition effect and pharmaceutical composition and application thereof Download PDF

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CN115716840A
CN115716840A CN202110971266.0A CN202110971266A CN115716840A CN 115716840 A CN115716840 A CN 115716840A CN 202110971266 A CN202110971266 A CN 202110971266A CN 115716840 A CN115716840 A CN 115716840A
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alkyl
radical
amino
pharmaceutically acceptable
stereoisomer
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吕贺军
陶志刚
叶文武
李坤
刘伟
纪海霞
刘军锋
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Shanghai Kunheng Medical Technology Co ltd
KPC Pharmaceuticals Inc
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Shanghai Kunheng Medical Technology Co ltd
KPC Pharmaceuticals Inc
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Abstract

The invention belongs to the field of medicines, and particularly relates to a heterocyclic compound with KRAS mutein inhibition effect, a pharmaceutical composition and application thereof, a pharmaceutical composition containing the compound or a stereoisomer and a pharmaceutically acceptable salt thereof, and the compound or the stereoisomer and the pharmaceutically acceptable salt thereof, which have inhibition effect on KRAS G12C and can be used as an anti-tumor effect.

Description

Heterocyclic compound with KRAS (KRAS-mutated protein) inhibition effect and pharmaceutical composition and application thereof
Technical Field
The invention relates to the field of medicines, in particular to a compound serving as a KRAS inhibitor, especially having an inhibiting effect on KRAS G12C, and more particularly, the compound provided by the invention can serve as an anti-tumor effect.
Background
The KRAS gene, collectively referred to as "Kirsten rat sarcoma virus oncogene homolog". The protein encoded by the KARS gene is a small gtpase and belongs to the RAS superfamily of super proteins. RAS is the most frequently mutated oncogene in human cancers, and mutation-induced RAS protein activation has been found in about 1/5 of all human tumors to date.
The RAS is positioned on the inner surface of a cell membrane, the upstream is Receptor Tyrosine Kinase (RTK), and after activation, downstream signal pathways such as PI3K, RAF and the like are regulated and controlled, so that the functions of growth, survival, migration, differentiation and the like of cells are regulated and controlled. RAS has two main states in the body: an inactive state in which it binds to the guanine dinucleotide GDP and an active state in which it binds to the guanine trinucleotide GTP. RAS activity is regulated by two proteins, a Guanosine Exchange Factor (GEF) promotes release of GDP from RAS proteins, allowing GTP binding to activate RAS; the Gtpase Activator Protein (GAP) activates the gtpase activity of the RAS protein, hydrolyzing GTP bound to the RAS protein to GDP, inactivating the RAS. Normally, RAS proteins are in an inactive state, and RAS is in a continuously active state after mutation, and downstream signaling pathways are continuously activated, thereby causing various cancers.
KRAS is the most frequently occurring subtype in the RAS family, with KRAS gene mutations accounting for 85% of the total RAS gene mutations (NRAS (12%) times, HRAS (3%) the least). In human cancers, KRAS gene mutations occur in nearly 90% of pancreatic cancers, 30-40% of colon cancers, 17% of endometrial cancers, and 15-20% of lung cancers (mostly NSCLC).
In 5 months 2021, AMG-510 (Sotoranib) from Amgen was approved by FDA for targeting drugs. However, it has been reported that resistance to the drug is observed in some patients taking AMG-510, and further studies have shown that the KRas (G12C) protein in some patients has new mutations after AMG-510, resulting in resistance to the AMG-510 drug. Therefore, the development of new safer and more effective KRas (G12C) targeted inhibitors still has urgent need and is of great significance.
The invention provides a novel KRAS G12C compound which shows exciting effect in the aspect of anti-tumor.
Disclosure of Invention
Accordingly, the present invention provides a KRAS inhibitor which is a heterocyclic compound having a tumor-inhibiting effect. Thus, the compounds of the present invention are useful for, but not limited to, the treatment of neoplastic diseases.
In order to achieve the above objects, the present invention provides a heterocyclic compound having the formula (I):
Figure BDA0003225850520000021
wherein the content of the first and second substances,
R 1 is C optionally substituted by 1 to 4 substituents 6-10 Aryl or 5-10 membered heteroaryl, said substituents being selected from: halogen, hydroxy, mercapto, amino, nitro, cyano, C 1-6 Alkyl radical, C 3-6 Cycloalkyl, C 1-6 Alkylamino radical, di (C) 1-6 Alkyl) amino, halo C 1-6 Alkyl, hydroxy C 1-6 Alkyl, amino C 1-6 Alkyl radical, C 1-6 Alkoxy radical, C 1-6 Alkylthio, halo C 1-6 Alkoxy, halo C 1-6 Alkylthio, hydroxy C 1-6 Alkoxy, hydroxy C 1-6 Alkylthio, amino C 1-6 Alkoxy or amino C 1-6 An alkylthio group;
R 2 is halogen, hydroxy, mercapto, amino, nitro, cyano, C 1-6 Alkyl radical, C 3-6 Cycloalkyl, C 1-6 Alkylamino radical, di (C) 1-6 Alkyl) amino, C 1-6 A haloalkyl group;
l is selected from-O-, -S-, -CONH-and-NH-;
y is N or CH;
m is 0, 1,2 or 3;
R 3 is selected from optionally substituted by 1-3R 3a Substituted 3-10 membered ringAlkyl or 3-10 membered heterocyclyl;
each R 3a Independently selected from: halogen, hydroxy, mercapto, amino, nitro, cyano, C 1-6 Alkyl radical, C 3-6 Cycloalkyl radical, C 1-6 Alkylamino radical, di (C) 1-6 Alkyl) amino, di (C) 1-6 Alkyl) amino-C 1-3 Alkyl radical, C 1-6 Haloalkyl, hydroxy C 1-6 Alkyl, amino C 1-6 Alkyl radical, C 1-6 Alkoxy radical, C 1-6 Alkylthio, halo C 1-6 Alkoxy radical, C 1-6 Halogenoalkylthio, hydroxy C 1-6 Alkoxy, hydroxy C 1-6 Alkylthio, amino C 1-6 Alkoxy or C 1-6 An aminoalkylthio group;
X 1 and X 2 Are each independently CH or N;
n is 0, 1,2 or 3;
each R is 4 Each independently is halogen, hydroxyl, sulfydryl, amino, nitro, cyano C 1-6 Alkyl radical, C 1-6 Alkyl radical, C 1-6 Alkylamino radical, di (C) 1-6 Alkyl) amino, halo C 1-6 Alkyl, hydroxy C 1-6 Alkyl, amino C 1-6 Alkyl, mercapto C 1-6 Alkyl radical, C 1-6 Alkoxy radical C 1-6 Alkyl or halo C 1-6 Alkoxy radical C 1-6 An alkyl group;
R 5 is-CO-CH = CHR 5a or-CO-CR 5a ＝CH 2 ，R 5a Each independently selected from hydrogen, amino, cyano, halogen, -CO-NH-C 1-6 Alkyl, -CO-N (C) 1-6 Alkyl radical) 2 、C 1-6 Alkyl radical, C 1-6 Alkoxy, halo C 1-6 Alkoxy radical, C 1-6 Alkylamino radical, di (C) 1-6 Alkyl) amino, halo C 1-6 Alkyl, hydroxy C 1-6 Alkyl or amino C 1-6 An alkyl group.
In some embodiments of the invention, the invention relates to a compound of formula (I) or a stereoisomer, pharmaceutically acceptable salt thereof, R 1 Is C optionally substituted with 1 to 4 substituents 6-10 Aryl or 5-to 10-memberedHeteroaryl, said substituents being selected from: halogen, hydroxy, mercapto, amino, nitro, cyano, C 1-6 Alkyl radical, C 1-6 Alkylamino radical, di (C) 1-6 Alkyl) amino, halo C 1-6 Alkyl, hydroxy C 1-6 Alkyl radical, C 1-6 Alkoxy, halo C 1-6 An alkoxy group.
In some embodiments of the invention, the invention relates to a compound of formula (I) or a stereoisomer, pharmaceutically acceptable salt thereof, R 1 Is a group optionally substituted with 1 to 4 substituents:
Figure BDA0003225850520000041
Figure BDA0003225850520000042
the substituent is selected from: halogen, hydroxy, amino, C 1-6 Alkyl, halo C 1-6 An alkyl group.
In some embodiments of the invention, the invention relates to a compound of formula (I) or a stereoisomer, pharmaceutically acceptable salt thereof, R 1 Is the following fragment:
Figure BDA0003225850520000043
in some embodiments of the present invention, the present invention relates to a compound of formula (I) or a stereoisomer, pharmaceutically acceptable salt thereof, wherein R 2 Methyl, ethyl, propyl and isopropyl.
In some embodiments of the present invention, the present invention relates to a compound of formula (I) or a stereoisomer, pharmaceutically acceptable salt thereof, wherein L is from-O-, -S-.
In some embodiments of the invention, the invention relates to a compound of formula (I) or a stereoisomer, pharmaceutically acceptable salt thereof, Y is N.
In some embodiments of the invention, the invention relates to a compound of formula (I) or a stereoisomer, pharmaceutically acceptable salt thereof, wherein m is 1.
In some embodiments of the invention, the invention relates to a compound of formula (I) or a stereoisomer, pharmaceutically acceptable salt thereof, R 3 Is selected from optionally substituted 1-3R 3a Substituted: cyclopropyl, cyclopentyl, cyclohexyl, tetrahydrofuryl, tetrahydrothienyl, tetrahydropyrrolyl, tetrahydroimidazolyl, tetrahydropyrazolyl, tetrahydrothiazolyl, tetrahydrooxazolyl, tetrahydropyranyl, piperidinyl, piperazinyl, morpholinyl, or hexahydropyrimidyl/cis-3-azabicyclo [3,3,0]Octane;
each R 3a Independently selected from: halogen, hydroxy, amino, cyano, C 1-6 Alkyl, N dimethylaminomethyl, C 1-6 A haloalkyl group.
In some embodiments of the invention, the invention relates to a compound of formula (I) or a stereoisomer, pharmaceutically acceptable salt thereof, R 3 The following fragments:
Figure BDA0003225850520000051
Figure BDA0003225850520000052
in some embodiments of the invention, the invention relates to a compound of formula (I) or a stereoisomer, pharmaceutically acceptable salt thereof, X 1 And X 2 Are all N.
In some embodiments of the invention, the invention relates to a compound of formula (I) or a stereoisomer, pharmaceutically acceptable salt thereof, n is 0, 1.
In some embodiments of the invention, the invention relates to a compound of formula (I) or a stereoisomer, pharmaceutically acceptable salt thereof, each R 4 Independently of each other, methyl and acetonitrile.
In some embodiments of the invention, the invention relates to a compound of formula (I) or a stereoisomer, pharmaceutically acceptable salt thereof, R 5 is-CO-CR 5a ＝CH 2 ，R 5a Selected from hydrogen and fluorine.
In some embodiments of the invention, the invention provides specific compounds or stereoisomers, pharmaceutically acceptable salts thereof,
Figure BDA0003225850520000053
Figure BDA0003225850520000061
all the above mentioned structural aspects are preferred embodiments of the respective aspects, respectively. Structural aspects related to the compound (I) portion of the present invention may be combined with each other as desired to obtain preferred compounds. Each combination represents and defines an individual embodiment or a general subset of the compounds (I) of the invention, and combinations of substituents and/or variables are permissible only if such combinations result in stable compounds or useful synthetic intermediates.
The present invention further relates to hydrates, solvates, polymorphs, metabolites, derivatives, isomers and prodrugs of the compounds of formula (I) (including all embodiments thereof).
The present invention further relates to pharmaceutically acceptable salts of the compounds of formula (I), including all embodiments thereof, including pharmaceutically acceptable salts of the compounds of formula (I), including all embodiments thereof, with inorganic or organic acids or bases. Pharmaceutically acceptable salts include, but are not limited to: salts with inorganic acids such as hydrochloride, phosphate, diphosphate, hydrobromide, sulfate, sulfinate, nitrate and the like; and salts with organic acids such as malate, maleate, fumarate, tartrate, succinate, citrate, acetate, lactate, methanesulfonate, p-toluenesulfonate, 2-isethionate, benzoate, salicylate, stearate, and alkanoates such as acetate, HOOC- (CH 2) n-COOH (where n is 0-4), and the like. Similarly, pharmaceutically acceptable cations include, but are not limited to: sodium, potassium, calcium, aluminum, lithium, and ammonium.
In addition, the pharmaceutically acceptable salts of the present invention can be synthesized from the parent compound containing an acid group or a base by conventional chemical methods. In general, such salts are prepared by the following method: prepared by reacting the free acid or base forms of these compounds with a stoichiometric amount of the appropriate base or acid in water or an organic solvent or a mixture of the two.
The compounds of the present invention may exist in specific geometric or stereoisomeric forms. The present invention contemplates all such compounds, including cis and trans isomers, (-) -and (+) -enantiomers, (R) -and (S) -enantiomers, diastereomers, (D) -isomers, (L) -isomers, as well as racemic and other mixtures thereof, such as enantiomerically or diastereomerically enriched mixtures, all of which are within the scope of the present invention. Additional asymmetric carbon atoms may be present in substituents such as alkyl groups. When they contain a plurality of asymmetric stereocenters, stereocombinations thereof and mixtures thereof are included within the scope of the invention, and alternative forms of the stereocompounds may be obtained by techniques conventional to those skilled in the art, e.g., by introduction of chiral starting materials, chiral resolution, and the like.
Cis and trans, or "Z" or "E" is used to indicate that cis and trans isomers exist in the compound, and generally, but not limited to, the difference in configuration involved in the moiety containing the double bond in the compound.
In another aspect, a compound as described herein may be formulated with a human-acceptable carrier into a pharmaceutical composition and administered to a mammalian host (such as a human patient) in a variety of forms suitable for the chosen route of administration, i.e., orally or parenterally, by intravenous, intramuscular, topical, transdermal, intrathecal, ocular, intranasal, intraperitoneal, or subcutaneous routes.
The compounds described herein may be administered systemically, e.g., orally or intravenously in combination with a pharmaceutically acceptable carrier, such as an inert diluent or an assimilable edible carrier. They may be enclosed in hard or soft shell gelatin capsules, may be compressed into tablets, or may be blended directly with the food of the patient's diet. For oral therapeutic administration, the active compound may be combined with one or more excipients and used in the form of ingestible tablets, buccal tablets, troches, capsules, elixirs, suspensions, syrups, wafers, and the like.
In another aspect, the present invention relates to a compound of formula (I) (including all embodiments thereof) or a pharmaceutical composition, which may be used for the preparation of a medicament for the treatment and/or prevention of cancer. In certain embodiments, the cancer is mediated by Kras mutation. In certain preferred embodiments, the Kras mutation is selected from one or more of Kras G12D, kras G12S, kras G12V and Kras G12C. In certain preferred embodiments, the Kras mutation is Kras G12C. In certain embodiments, the cancer is selected from lung cancer, colorectal cancer, or pancreatic cancer. In certain preferred embodiments, the lung cancer is selected from non-small cell lung cancer. The effective amount refers to the dosage of the medicine capable of preventing, alleviating, delaying, inhibiting or curing the symptoms of the subject. The size of the administered dose is determined by the drug administration mode, the pharmacokinetics of the agent, the severity of the disease, the individual signs of the subject (sex, weight, height, age), and the like.
In another aspect, the invention relates to a kit comprising a compound of the invention, or a pharmaceutically acceptable salt thereof, and instructions for administering the compound, or a pharmaceutically acceptable salt thereof, to a subject having cancer.
The starting materials for the following reactions are generally known compounds or can be prepared by known procedures or obvious modifications thereof.
The various starting materials, intermediates and compounds described herein can be isolated and purified, where appropriate, using conventional techniques such as precipitation, filtration, crystallization, evaporation, distillation and chromatography. Characterization of these compounds can be performed using conventional methods, such as by melting point, mass spectrometry, nuclear magnetic resonance, and various other spectroscopic analyses.
Some examples of compounds of the present invention can be realized by the following reaction scheme:
general reaction scheme 1:
Figure BDA0003225850520000091
wherein R is 1 、R 2 、R 3 、R 4 、R 5 And L is as defined for formula (I). In step A, compound A1 is subjected to bromination or the like to give compound A2, which is carried out using a suitable brominating agent (e.g., liquid bromine) in a suitable solvent (e.g., acetic acid). In step B, compound A2 is reacted with benzoyl isocyanate to give benzoylthiourea (compound A4), which can be carried out by heating in a suitable aprotic solvent (e.g., 1, 4-dioxane). In step C, compound A4 is ring-closed in the presence of a suitable base (e.g., sodium hydroxide) in a suitable solvent (e.g., methanol) to provide compound A5. In step D, compound A5 is reacted with a suitable methylating agent (e.g., methyl iodide) to provide compound A6. In step E, compound A6 is reacted with a suitable chlorinating agent, such as phosphorus oxychloride, to give compound A7, which is usually carried out under heating. In step F, compound A7 undergoes a substitution reaction with piperazine (compound A8) to provide compound A9. In step G, compound A9 is reacted with a suitable protecting reagent (di-tert-butyl dicarbonate) to give compound A10. In step H, compound A10 is reacted with a suitable boron reagent (such as pinacolborane) in the presence of a catalyst (such as tris (dibenzylideneacetone) dipalladium) and a suitable ligand (such as 2-dicyclohexylphosphine-2 ',4',6' -triisopropylbiphenyl) to provide compound A11. In step I, compound A11 is reacted with a halide R in the presence of a catalyst, such as tris (dibenzylideneacetone) dipalladium, and a suitable ligand, such as 2-dicyclohexylphosphine-2 ',4',6' -triisopropylbiphenyl 1 Coupling of-X (e.g., pinacolborane) affords compound A12. In step J, compound A12 is reacted with a suitable oxidizing agent (e.g., m-chlorobenzoic acid) to provide sulfoxide A13. In step K, compound A13 is reacted with a nucleophile R in the presence of a suitable base (e.g.sodium tert-butoxide) in a suitable solvent (e.g.toluene) 3 -(CH 2 ) m Reaction of-L gives compound A14. In step L, compound A14 is in a suitable acid (e.g., trifluoroacetic acid)The protecting group is removed to obtain a compound A15. In step M, compound A15 is reacted with an acid chloride reagent in the presence of a suitable base (e.g., triethylamine) or condensed with the corresponding acid in the presence of a suitable condensing agent (e.g., N, N, N ', N' -tetramethyl-O- (7-azabenzotriazol-1-yl) urea hexafluorophosphate) to provide the compound of formula (I).
Reaction scheme 2:
Figure BDA0003225850520000111
wherein R is 1 、R 2 、R 3 、R 4 、R 5 And L is as defined for formula (I). Preparation of compounds B2 to B10 the preparation of A2 to A10 in the general scheme 1. Compound B10 is reacted with a suitable boronic acid (e.g., R-B (OH)) in the presence of a suitable metal catalyst 2 ) Or trialkyltin (e.g. R-Sn (n-Bu) 3 ) A coupling reaction takes place to give compound B12. In step I, compound B12 is reacted with a suitable oxidizing agent (e.g., m-chlorobenzoic acid) to provide sulfoxide B13. In step J, compound B13 is reacted with nucleophile R in the presence of a suitable base (e.g., sodium tert-butoxide) in a suitable solvent (e.g., toluene) 3 -(CH 2 ) m Reaction of-L gives compound B14. In step K, compound B14 is deprotected in a suitable acid (e.g., trifluoroacetic acid) to provide compound B15. In step L, compound B15 is reacted with an acid chloride reagent in the presence of a suitable base (e.g., triethylamine) or condensed with the corresponding acid in the presence of a suitable condensing agent (e.g., N, N, N ', N' -tetramethyl-O- (7-azabenzotriazol-1-yl) urea hexafluorophosphate) to provide the compound of formula (I).
While certain embodiments have been illustrated and described, it will be appreciated that changes and modifications may be made therein in accordance with ordinary skill in the art without departing from the technology as defined in the following claims in their broader aspects.
The invention has the following beneficial effects: the compound of the invention shows better inhibitory activity for inhibiting conversion of KRas (G12C) -GDP to KRas (G12C) -GTP mediated by SOS1, and part of the compound shows equivalent or better inhibitory activity than that of a control drug AMG-510. This indicates that the compounds of the invention are expected to be useful in the treatment of diseases caused/associated in particular by KRas G12C mutations.
Noun interpretation
"optional" or "optionally" means that the subsequently described event or circumstance may or may not occur, and that the description includes instances where the event or circumstance occurs and instances where it does not. For example, "optionally substituted alkyl" includes "alkyl" and "substituted alkyl" as defined herein. It will be understood by those skilled in the art that for any group containing one or more substituents, such groups are not intended to introduce any substitution or substitution pattern that is sterically impractical, synthetically non-feasible and/or inherently unstable.
"alkyl" includes straight and branched chains having the indicated number of carbon atoms (typically 1 to 20 carbon atoms, for example 1 to 8 carbon atoms, such as 1 to 6 carbon atoms). E.g. C 1-6 Alkyl groups include straight and branched chain alkyl groups of 1 to 6 carbon atoms. Examples of alkyl groups include, but are not limited to, methyl, ethyl, propyl, isopropyl, n-butyl, sec-butyl, tert-butyl, pentyl, 2-pentyl, isopentyl, neopentyl, hexyl, 2-hexyl, 3-methylpentyl, and the like. Alkylene is another subset of alkyl and refers to the same residue as alkyl, but having two points of attachment, and when it is stated herein that certain alkyl groups have two points of attachment, it is also meant alkylene. The alkylene group typically has 2 to 20 carbon atoms, for example 2 to 8 carbon atoms, such as 2 to 6 carbon atoms. When naming alkyl residues with a specific carbon number, all geometric isomers with that carbon number are intended to be included, e.g., "butyl" is intended to include n-butyl, sec-butyl, isobutyl, and tert-butyl; "propyl" includes n-propyl and isopropyl. "lower alkyl" means an alkyl group having 1 to 4 carbons.
"alkenyl" means having the indicated number of carbon atoms (typically 1 to 8 carbon atoms, e.g., 2 to 4 carbon atoms) and at least 1 and preferably 1 to 2 vinyl groups (>C＝C<) A linear or branched hydrocarbon radical of an unsaturated site. Examples of such groups are, for example, vinyl, allyl and but-3-en-1-yl.Included within this term are the cis and trans isomers or mixtures of these isomers. "lower alkenyl" means an alkenyl group having 1-4 carbons, which may pass through C 2-4 Alkenyl groups.
Haloalkyl (haloalkenyl, haloalkynyl) groups are all derived from alkyl (alkenyl, alkynyl) groups as previously defined by replacing one or more hydrogen atoms of the hydrocarbon chain with halogen atoms which may be the same or different, independently of one another. If the haloalkyl (haloalkenyl, haloalkynyl) is to be further substituted, the substitution can be carried out in each case independently of one another on all of the carbon atoms bearing the hydrogen in monosubstituted or polysubstituted form. Examples of haloalkyl (haloalkenyl, haloalkynyl) are-CF 3 、-CHF 2 、-CH 2 F、-CF 2 CF 3 -CHFCF 3 、-CH 2 CF 3 、-CF 2 CH 3 、-CHFCH 3 、-CF 2 CF 2 CF 3 、-CF 2 CH 2 CH 3 、-CF＝CF 2 、-CCl＝CH 2 、-CBr＝CH 2 、-C≡CCF 3 、-CHFCH 2 CH 3 、-CHFCH 2 CF 3 And the like.
"cycloalkyl" means a non-aromatic, partially saturated, or fully saturated, carbocyclic ring having the specified number of carbon ring atoms (e.g., 3 to 10, or 3 to 8, or 3 to 6 ring carbon atoms). Cycloalkyl groups can be monocyclic or polycyclic (e.g., bicyclic, tricyclic). Examples of cycloalkyl groups include cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl and cyclohexyl, as well as bridged and caged ring groups (e.g., bicyclo [2.2.2 ]]Octane). Lower cycloalkanes are in general C 3-6 The monocyclic ring, lower cycloalkyl in general, may be preferably a fully saturated carbocyclic ring, unless otherwise specified.
"hydroxyalkyl (hydroxycycloalkyl)" includes the substitution of one or more hydroxyl groups for the above-described straight and branched alkyl (cycloalkyl) groups having the indicated number of carbon atoms (typically 1 to 20 carbon atoms, e.g., 1 to 8 carbon atoms, such as 1 to 6 carbon atoms), as combinations of substituents and/or variables are permissible only if such combinations result in stable compounds or useful synthetic intermediates.
"halo-hydroxyalkyl" includes the substitution of one or more hydroxyl groups and halogens for the above-described straight and branched chain alkyl groups having the indicated number of carbon atoms (typically 1 to 20 carbon atoms, e.g., 1 to 8 carbon atoms, such as 1 to 6 carbon atoms), as combinations of substituents and/or variables are permissible only if such combinations result in stable compounds or useful synthetic intermediates.
"aryl" means an aromatic carbocyclic ring having the specified number of carbon atoms in the ring (e.g., 6 to 12 or 6 to 10 carbon atoms). The aryl group can be monocyclic or polycyclic (e.g., bicyclic, tricyclic). In some cases, both rings of the polycyclic aryl are aromatic (e.g., naphthyl). In other instances, the polycyclic aryl can include a non-aromatic ring fused to an aromatic ring (e.g., cycloalkyl, cycloalkenyl, heterocycloalkyl, heterocycloalkenyl) provided that the polycyclic aryl is bonded to the parent structure through an atom in the aromatic ring. Thus, a1, 2,3, 4-tetrahydronaphthalen-5-yl group (in which the moiety is bonded to the parent structure via an aromatic carbon atom) is considered an aryl group, while a1, 2,3, 4-tetrahydronaphthalen-1-yl group (in which the moiety is bonded to the parent structure via a non-aromatic carbon atom) is not considered an aryl group. Similarly, 1,2,3, 4-tetrahydroquinolin-8-yl (where the moiety is bonded to the parent structure via an aromatic carbon atom) is considered an aryl group, and 1,2,3, 4-tetrahydroquinolin-1-yl (where the moiety is bonded to the parent structure via a non-aromatic nitrogen atom) is not considered an aryl group. However, the term "aryl" does not include or overlap with "heteroaryl" as defined herein, regardless of the point of attachment (e.g., both quinolin-5-yl and quinolin-2-yl are heteroaryl). In some cases, where not otherwise specified, the aryl ring may be further substituted with functional groups well known in the art, without affecting the definition of the number of carbon atoms on the aryl ring. In some cases, aryl is phenyl or naphthyl. In some cases, aryl is phenyl. Other examples of aryl groups comprising an aromatic carbocyclic ring fused to a non-aromatic ring are described below. C as described herein 0 -C 1 alkylene-C 6-10 Aryl, typically when C 0 alkylene-C 6-10 When aryl is said to correspond to aryl, C 1 alkylene-C 6-10 By aryl is generally meant an aryl group which is adjacent to a methylene group, such as benzyl.
"heteroaryl" means an aromatic ring (e.g., a 5-12 or 5-10 membered heteroaryl) containing the indicated number of ring atoms, which ring atoms are composed of one or more heteroatoms (e.g., 1,2,3, or 4 heteroatoms) selected from N, O, and S, and the remaining ring atoms are carbon. A 5-membered heteroaryl is a heteroaryl having 5 ring atoms. A 6-membered heteroaryl is a heteroaryl having 6 ring atoms. In some embodiments, the total number of S and O atoms in the heteroaryl group is no more than 2. In some embodiments, the total number of S and O atoms in the heteroaryl group is no more than 1. Unless otherwise indicated, heteroaryl groups may be bonded to the parent structure through a carbon or nitrogen atom, as valency permits. For example, "pyridyl" includes 2-pyridyl, 3-pyridyl, and 4-pyridyl, and "pyrrolyl" includes 1-pyrrolyl, 2-pyrrolyl, and 3-pyrrolyl. When a nitrogen is present in a heteroaryl ring, the nitrogen may be present in the oxidation state (i.e., N + -O-), as the nature of the adjacent atoms and groups permits. Further, when sulfur is present in a heteroaryl ring, the sulfur may be in the oxidation state (i.e., S + -O-or SO) as the nature of adjacent atoms and groups permits 2 ) Are present. Heteroaryl groups can be monocyclic or polycyclic (e.g., bicyclic, tricyclic). In some cases, the heteroaryl group is monocyclic. Examples include pyrrole, pyrazole, imidazole, triazole (e.g., 1,2, 3-triazole, 1,2, 4-triazole), tetrazole, furan, isoxazole, oxazole, oxadiazole (e.g., 1,2, 3-oxadiazole, 1,2, 4-oxadiazole, 1,3, 4-oxadiazole), thiophene, isothiazole, thiazole, thiadiazole (e.g., 1,2, 3-thiadiazole, 1,2, 4-thiadiazole, 1,3, 4-thiadiazole), pyridine, pyridazine, pyrimidine, pyrazine, triazine (e.g., 1,2, 4-triazine, 1,3, 5-triazine), and tetrazine. In other instances, the polycyclic heteroaryl can include a non-aromatic ring (e.g., cycloalkyl, cycloalkenyl, heterocycloalkyl, heterocycloalkenyl) fused to a heteroaryl ring, so long as the polycyclic heteroaryl is bonded to the parent structure through an atom in the aromatic ring. For example, 4,5,6,7-tetrahydrobenzo [ d ]]Thiazol-2-yl (wherein the moiety is bonded to the parent bond via an aromatic carbon atom)Structure) is considered to be heteroaryl, and 4,5,6, 7-tetrahydrobenzo [ d ]]Thiazol-5-yl (wherein the moiety is bonded to the parent structure via a non-aromatic carbon atom) is not considered heteroaryl. Examples of polycyclic heteroaryls consisting of a heteroaryl ring fused to a non-aromatic ring are described below.
"heterocycloalkyl" means a non-aromatic, partially or fully saturated ring (e.g., 3-10 or 3-7 membered heterocycloalkyl), sometimes referred to herein as heterocyclyl, having the indicated number of ring atoms, which ring atoms are composed of one or more heteroatoms (e.g., 1,2,3, or 4 heteroatoms) selected from N, O, and S, and the remaining ring atoms are carbon. A 5-membered heterocycloalkyl group is a heterocycloalkyl group having 5 ring atoms. A 6-membered heterocycloalkyl group is a heterocycloalkyl group having 6 ring atoms. The heterocycloalkyl group can be monocyclic or polycyclic (e.g., bicyclic, tricyclic). Examples of heterocycloalkyl groups include oxacyclopropyl, aziridinyl, azetidinyl, pyrrolidinyl, imidazolidinyl, pyrazolidinyl, piperidinyl, piperazinyl, morpholinyl, and thiomorpholinyl. When nitrogen is present in the heterocycloalkyl ring, the nitrogen may be present in the oxidation state (i.e., N + -O-) as the nature of the adjacent atoms and groups permits. Examples include piperidinyl-N-oxide and morpholinyl-N-oxide. Further, when sulfur is present in the heterocycloalkyl ring, the sulfur may be in the oxidized state (i.e., S + -O-or-SO) as the nature of the adjacent atoms and groups permits 2 -) are present. Examples include thiomorpholine S-oxide and thiomorpholine S, S-dioxide. In addition, one ring of the polycyclic heterocycloalkyl can be aromatic (e.g., aryl or heteroaryl) so long as the polycyclic heterocycloalkyl is bonded to the parent structure via a non-aromatic carbon or nitrogen atom. For example, 1,2,3, 4-tetrahydroquinolin-1-yl (where the moiety is bonded to the parent structure via a non-aromatic nitrogen atom) is considered a heterocycloalkyl group, while 1,2,3, 4-tetrahydroquinolin-8-yl (where the moiety is bonded to the parent structure via an aromatic carbon atom) is not considered a heterocycloalkyl group. Lower heterocycloalkanes in general are C 3-6 The monocyclic ring, lower heterocycloalkyl group and the like may be preferably a fully saturated carbocyclic ring in general, unless otherwise specified.
"alkoxy" refers to an alkyl group of the indicated number of carbon atoms attached through an oxygen bridge, such as methoxy, ethoxy, propoxy, isopropoxy, n-butoxy, sec-butoxy, tert-butoxy, pentyloxy, 2-pentyloxy, isopentyloxy, neopentyloxy, hexyloxy, 2-hexyloxy, 3-methylpentyloxy, and the like. Alkoxy is also intended to include cycloalkyl groups as defined above, also attached through an oxygen bridge. Alkoxy groups typically have 1-6 carbon atoms connected by an oxygen bridge. "lower alkoxy" means an alkoxy group having 1 to 4 carbons.
The term "halo" includes fluoro, chloro, bromo and iodo.
The term "substituted" as used herein means that any one or more hydrogens on the designated atom or group is replaced with a selection from the designated group, provided that the designated atom's normal valence is not exceeded. When the substituent is oxo (i.e., = O), then 2 hydrogens on the atom are replaced. Combinations of substituents and/or variables are permissible only if such combinations result in stable compounds or useful synthetic intermediates. A stable compound or stable structure is intended to imply that the compound is sufficiently robust to survive isolation from a reaction mixture and subsequent formulation into an agent of at least practical utility. Unless otherwise indicated, substituents are named into the core structure. For example, it will be understood that when (cycloalkyl) alkyl is listed as a possible substituent, the point of attachment of that substituent to the core structure is in the alkyl moiety.
Detailed Description
To further illustrate the present invention, the compounds provided herein, as well as methods of making and using the same, are described in detail below with reference to the following examples, which are not intended to be limiting. It will be apparent to those skilled in the art that various changes and modifications can be made in the specific embodiments of the invention without departing from the spirit and scope of the invention. The examples, in which specific conditions are not specified, were conducted under conventional conditions or conditions recommended by the manufacturer. The reagents or instruments used are not indicated by the manufacturer, and are all conventional products commercially available.
Example 1: synthesis of intermediate INT 1
Figure BDA0003225850520000171
Intermediate INT 1 was prepared by the following synthetic route:
Figure BDA0003225850520000172
synthesis of INT 1-2: n is a radical of hydrogen 2 6-chloro-4-methylpyridin-2-amine (4.8g, 33.7 mmol) was dissolved in N, N-dimethylformamide (80 mL) in a three-necked flask with an internal temperature thermometer under protection. After cooling to 0 ℃ and adding sodium hydride (4.2g, 60%,101.1 mmol) in portions, maintaining 0 ℃ and stirring for 30 minutes, p-methoxybenzyl chloride (12.7g, 77.4mmol) was dissolved in N, N-dimethylformamide (10 mL) and added dropwise to the above reaction system. After the addition was complete, the temperature was raised to room temperature, the reaction was stirred for 12h and monitored by LC-MS for completion. Water (100 mL) was added for dilution, and extraction was performed with ethyl acetate (100 mL × 2), liquid separation was performed, the organic phases were combined, the organic phase was washed twice with saturated brine (30 mL), dried over anhydrous sodium sulfate, the solid was removed by filtration, the filtrate was concentrated to dryness under reduced pressure, the residue was purified by Biotage Flash (petroleum ether: ethyl acetate,
Figure BDA0003225850520000182
Figure BDA0003225850520000181
) To give INT 1-2-6-chloro-N, N-bis (4-methoxybenzyl) -4-methylpyridin-2-amine as a white solid (8.7 g, yield: 68.1%).
MS(ESI):m/z＝383.1[M+1] + 。
Synthesis of INT 1-3: n is a radical of 2 INT 1-2-chloro-N, N-bis (4-methoxybenzyl) -4-methylpyridin-2-amine (8.5g, 22.3mmol) was dissolved in acetonitrile (100 mL) under protection in a three-necked flask with an internal temperature thermometer, and iodosuccinimide (23g, 133.8mmol) was slowly added. After the addition of the water is finished,the nitrogen was replaced 3 times, the reaction was stirred for 12h at 80 ℃ and the completion of the reaction was monitored by LCMS. The reaction solution was concentrated under reduced pressure to dryness, the residue was dissolved in ethyl acetate (100 mL), washed twice with a saturated aqueous solution of sodium hydrogencarbonate (30 mL), once with a saturated saline solution (30 mL), dried over anhydrous sodium sulfate, filtered to remove solids, the filtrate was concentrated under reduced pressure to dryness, and the residue was purified by Biotage Flash (petroleum ether: ethyl acetate,
Figure BDA0003225850520000183
) To give INT 1-3-6-chloro-5-iodo-N, N-bis (4-methoxybenzyl) -4-methylpyridin-2-amine as a yellow solid (4.7 g, yield: 42.2%).
MS(ESI):m/z＝509.1[M+1] + 。
INT 1 Synthesis: n is a radical of 2 INT 1-3-chloro-5-iodo-N, N-bis (4-methoxybenzyl) -4-methylpyridin-2-amine (2.0 g, 3.94mmol) was dissolved in N, N-dimethylformamide (20 mL) under protection in a three-necked flask with an internal temperature thermometer, and methyl 2, 2-difluoro-2- (fluorosulfonyl) acetate (1.1g, 5.9mmol) and cuprous iodide (1.1g, 5.9mmol) were slowly added. After the addition was complete, nitrogen was replaced 3 times, the reaction was stirred for 4h at 100 ℃ and the completion of the reaction was monitored by LCMS. Water (20 mL) was added to dilute and extracted with ethyl acetate (20 mL × 2), the liquids were separated, the organic phases were combined, the organic phase was washed twice with saturated brine (30 mL), dried over anhydrous sodium sulfate, the solid was removed by filtration, the filtrate was concentrated to dryness under reduced pressure, the residue was purified by Biotage Flash (petroleum ether: ethyl acetate,
Figure BDA0003225850520000193
) To give INT 1 6-chloro-N, N-bis (4-methoxybenzyl) -4-methyl-5- (trifluoromethyl) pyridin-2-amine as a white solid (1.1 g, yield: 62.3%).
MS(ESI):m/z＝451.2[M+1] + 。
Example 2: synthesis of intermediate INT 2
Figure BDA0003225850520000191
Intermediate INT 2 was prepared by the following synthetic route:
Figure BDA0003225850520000192
INT 2-2 Synthesis: n is a radical of 2 Under protection, INT 2-1-3-amino-4-methylthiophene-2-carboxylic acid methyl ester (20g, 110mmol) is dissolved in dichloromethane (200 mL) in a three-necked flask with an internal temperature thermometer, then the temperature is reduced to 0 ℃ by using an ice water bath, glacial acetic acid (22 mL) and bromine (12.8 mL) are slowly added, after the addition is finished, the mixture is heated to 50 ℃ for reaction for 12h, and LCMS monitors that the reaction is finished. The reaction was quenched with 200mL of water, extracted with dichloromethane (200 mL. Times.2), the organic phases were combined, washed twice with saturated brine (30 mL), dried over anhydrous sodium sulfate, filtered to remove solids, the filtrate was concentrated to dryness under reduced pressure, and the residue was purified by Biotage Flash (petroleum ether: ethyl acetate,
Figure BDA0003225850520000194
) To give methyl INT 2-3-amino-5-bromo-4-methylthiophene-2-carboxylate (7.80 g, yield: 26.6%).
MS(ESI):m/z＝251.9[M+1] + 。
INT 2-3 Synthesis: n is a radical of 2 INT 2-3-amino-5-bromo-4-methylthiophene-2-carboxylic acid methyl ester (7.8g, 31.2mmol) was dissolved in dioxane (100 mL) under protection in a three-necked flask with an internal temperature thermometer, followed by slow addition of benzoyl isothiocyanate (6.1g, 37.4 mmol). After the addition, the mixture was heated to 110 ℃ and stirred for 4 hours. The reaction was monitored by LCMS for completion. The reaction solution was concentrated to dryness under reduced pressure, and the residue was purified by Biotage Flash (petroleum ether: ethyl acetate,
Figure BDA0003225850520000201
) To give INT 2-3 (E) -5-bromo-4-methyl-3- (3- (2-methyl-2-alkenyl) thioureido) thiophene-2-carboxylic acid as a brown solid (10.1 g, yield: 81.1%).
MS(ESI):m/z＝413.0[M+1] + 。
INT 2-4 Synthesis: INT 2-3 (E) -5-bromo-4-methyl-3- (3- (2-methyl-2-alkenyl) thioureido) thiophene-2-carboxylic acid (10.1g, 24.5 mmol) was dissolved in methanol (50 mL) in a three-necked flask with an internal temperature thermometer, followed by slow addition of sodium hydroxide (3g, 73.5 mmol). After the addition, the reaction was stirred at room temperature for 12 hours. The reaction was monitored by LCMS for completion. Methanol was removed by concentration under reduced pressure, and the residue was adjusted to pH =6 with 2N hydrochloric acid, stirred for 30 minutes, and filtered to give INT 2-4-6-bromo-2-mercapto-7-methylthiophene [3,2-d ] pyrimidin-4 (3H) -one (4.3 g, yield: 62.5%) as a pale yellow solid.
MS(ESI):m/z＝276.9[M+1] + 。
INT 2 synthesis: INT 2-4-bromo-2-mercapto-7-methylthiophene [3,2-d ] pyrimidin-4 (3H) -one (1g, 3.6 mmol) was dissolved in N, N-dimethylformamide (20 mL) in a three-necked flask with an internal thermometer, followed by slow addition of sodium acetate (290mg, 3.6 mmol), stirring at room temperature for 30 minutes, and addition of iodomethane (512mg, 3.6 mmol). After the addition, the reaction was stirred at room temperature for 2 hours. Completion of the reaction was monitored by LCMS. Water (20 mL) was added, stirred at room temperature for 20 minutes, and filtered to give INT 2-bromo-7-methyl-2- (methylthio) thieno [3,2-d ] pyrimidin-4 (3H) -one as a brown solid (1.01 g, yield: 96.1%).
MS(ESI):m/z＝290.9[M+1] + 。
Example 3: synthesis of intermediate INT 3
Figure BDA0003225850520000211
Intermediate INT 3 was prepared by the following synthetic route:
Figure BDA0003225850520000212
INT 3-1 Synthesis: n is a radical of 2 INT 2-bromo-7-methyl-2- (methylthio) thieno [3,2-d ] in a three-necked flask with internal thermometer under protection]Pyrimidin-4 (3H) -one (1g, 3.4 mmol) was dissolved in phosphorus oxychloride (20 mL), followed by slow addition of pyridine (830mg, 10.3 mmol), and after the addition, the reaction was stirred at 120 ℃ for 3H. The completion of the reaction was monitored by LC-MS. Concentrated to dryness under reduced pressure, and the residue was dissolved in methylene chloride (100 mL)Washed twice with water (100 mL), once with brine (100 mL), dried over anhydrous sodium sulfate, filtered to remove solids, and the filtrate concentrated to dryness under reduced pressure to give INT 3-1-bromo-4-chloro-7-methyl-2- (methylthio) thieno [3,2-d ] as a yellow solid]Pyrimidine (1.1 g, yield: 95.2%).
MS(ESI):m/z＝310.8[M+1] + 。
Synthesis of INT 3-2: INT 3-1-bromo-4-chloro-7-methyl-2- (methylthio) thieno [3,2-d ] pyrimidine (1g, 3.2 mmol) was dissolved in dioxane (20 mL) in a three-necked flask with an internal temperature thermometer, and then (S) -2- (piperazin-2-yl) acetonitrile (1.9g, 6.4mmol), N, N-diisopropylethylamine (1.2g, 9.7mmol) were slowly added. After the addition, the mixture was heated to 70 ℃ and stirred for 5 hours. The completion of the reaction was monitored by LC-MS. Direct concentration under reduced pressure gave INT 3-2 (S) -2- (4- (6-bromo-7-methyl-2- (methylthio) thieno [3,2-d ] pyrimidin-4-yl) piperazin-2-yl) acetonitrile (1.3 g, yield: 95.5%) as a pale yellow solid.
MS(ESI):m/z＝398.0[M+1] + 。
INT 3 synthesis: INT 3-2 (S) -2- (4- (6-bromo-7-methyl-2- (methylthio) thieno [3, 2-d) in a three-necked flask with an internal temperature thermometer]Pyrimidin-4-yl) piperazin-2-yl) acetonitrile (1.3 g,1.2 mmol) was dissolved in dioxane (10 mL), N-diisopropylethylamine (1.2 g,2 mmol) and 4-dimethylaminopyridine (35mg, 0.10mmol) were added, and di-tert-butyl dicarbonate (2.6 g, 120.1mmol) was slowly added. After the addition, the mixture was heated to 40 ℃ and stirred for 2 hours. The completion of the reaction was monitored by LC-MS. The reaction was then quenched with 200mL of water, extracted with dichloromethane (200 mL. Times.2), the organic phases were combined, washed twice with saturated brine (30 mL), dried over anhydrous sodium sulfate, filtered to remove solids, the filtrate was concentrated to dryness under reduced pressure, and the residue was purified by Biotage Flash (petroleum ether: ethyl acetate,
Figure BDA0003225850520000221
) To give INT 3 (S) -4- (6-bromo-7-methyl-2- (methylthio) thieno [3,2-d ] as a brown solid]Pyrimidin-4-yl) -2- (cyanomethyl) piperazine-1-carboxylic acid tert-butyl ester (1.5 g, yield: 95.1%).
MS(ESI):m/z＝498.1[M+1] + 。
Example 4: synthesis of intermediate INT 4
Figure BDA0003225850520000222
INT 4 can be obtained by a synthesis method similar to INT 3 and by using (S) -1-N-tert-butyloxycarbonyl-2-methylpiperazine as a starting material
LCMS m/z(ESI)：473.1[M+1] + 。
Example 5: synthesis of intermediate INT 5
Figure BDA0003225850520000223
INT 5 can be obtained by a synthetic method similar to INT 3 and using (S) -4-N-tert-butoxycarbonyl-2-methylpiperazine as a starting material
LCMS m/z(ESI)：473.1[M+1] + 。
Example 6: synthesis of intermediate INT 6
Figure BDA0003225850520000231
Intermediate INT 6 was prepared by the following synthetic route:
Figure BDA0003225850520000232
INT 6-1 Synthesis: n is a radical of hydrogen 2 INT 3 (1.00g, 2.01mmol), pinacolborane (5.14g, 40.20mmol), triethylamine (0.61g, 6.03mmol), 2-dicyclohexylphosphine-2 ',4',6' -triisopropylbiphenyl (0.19g, 0.40mmol) and tris (dibenzylideneacetone) dipalladium (0.18g, 0.20mmol) were added to dioxane (16.0 ml) under protection, and stirring was carried out at 105 ℃ for 0.5 hour, and LC-MS monitored for the absence of a raw material. The reaction solution was used in the next step without purification.
MS(ESI):m/z＝464.2[M+H] + 。
INT 6-2 Synthesis: n is a radical of 2 Under protection, water (4.0 ml), potassium carbonate (0.83g, 6.03mmol), INT 1 (0.54g, 1.21mmol) and [1,1' -bis (diphenylphosphino) ferrocene are added into the reaction solution in the previous step]Palladium dichloride (0.15g, 0.20mmol), stirred at 105 ℃ for 2 hours, and LC-MS monitored that the reaction was complete. The reaction solution was filtered, the filter cake was washed with dichloromethane (50 ml × 3), the filtrate was collected, concentrated to dryness, and the crude product was purified by Biotage Flash (ethyl acetate: petroleum ether = 0-20%) to give INT 6-2 (170.0 mg, two-step yield: 10%, purity: 93%) as a pale yellow solid.
MS(ESI):m/z＝834.3[M+H] + 。
INT 6 synthesis: n is a radical of 2 INT 6-2 (0.16g, 0.18mmol) was dissolved in dichloromethane (10.0 ml) in a three-necked flask with an internal temperature thermometer under protection, m-chloroperoxybenzoic acid (0.03g, 0.18mmol) was added at 0 ℃ and stirred at 0 ℃ for 1 hour, and LC-MS monitored the completion of the reaction. The reaction was quenched with water (20 ml), extracted with dichloromethane (30 ml × 3), the organic phases combined, washed with saturated brine (30 ml), dried over anhydrous sodium sulfate, filtered to remove the sodium sulfate, and the filtrate concentrated under reduced pressure to give INT 6 (0.17 g, crude) as a yellow solid.
MS(ESI):m/z＝850.3[M+H] + 。
Example 7: synthesis of Compound 1
Figure BDA0003225850520000241
Compound 1 is prepared by the following synthetic route
Figure BDA0003225850520000242
Synthesis of Compound 1-1: n is a radical of hydrogen 2 2-Fluorohexahydro-1H-pyrrolizin-7A-yl) methanol (85.9mg, 0.54mmol) was dissolved in toluene (2.0 ml) in a three-necked flask with an internal temperature thermometer under protection, sodium t-butoxide (51.8mg, 0.54mmol), INT 6 (155.0mg, 0.18mmol) was added at 0 ℃ and stirred at 0 ℃ for 1 hour, and the reaction was monitored by LC-MS for completion. The reaction solution was quenched with water (20 ml)Then, the mixture was extracted with ethyl acetate (30 ml. Times.3), the organic phases were combined, washed with saturated brine (30 ml), dried over anhydrous sodium sulfate, filtered to remove sodium sulfate, and the filtrate was concentrated under reduced pressure to give compound 1-1 (310.0 mg, crude product) as a yellow solid.
MS(ESI):m/z＝945.4[M+H] + 。
Synthesis of Compounds 1-2: compound 1-1 (310.0 mg, 0.33mmol) was dissolved in trifluoroacetic acid (10.0 ml) in a three-necked flask with an internal temperature thermometer, stirred at 80 ℃ for 2 hours, and the reaction was monitored by LC-MS for completion. The reaction solution was concentrated under reduced pressure, and the crude product was purified by Biotage Flash (methanol: dichloromethane = 0-10%) to obtain compound 1-2 as a yellow solid (100.0 mg, yield: 46%, purity: 93%).
MS(ESI):m/z＝605.2[M+H] + 。
Synthesis of Compound 1: n is a radical of 2 Compound 1-2 (45.0 mg, 0.07mmol) was dissolved in dichloromethane (3.0 ml) in a three-necked flask equipped with an internal temperature thermometer under protection, triethylamine (21.21mg, 0.21mmol) and acryloyl chloride (6.3mg, 0.07mmol) were added at 0 ℃, and the reaction was stirred at 0 ℃ for 0.5 hour and monitored by LC MS for completion. The reaction solution was quenched with methanol (5 ml), concentrated under reduced pressure, and the crude product was purified by Prep-HPLC (acetonitrile/TFA aq (0.01%)) to give compound 1 as a white solid (11.8 mg, yield: 25%, purity: 98%).
MS(ESI):m/z＝659.2[M+H] + ；
1 H NMR(400MHz,Methanol-d4):δ6.91-6.75(m,1H),6.69(s,1H),6.39-6.25(m,1H),5.94-5.80(m,1H),5.69-5.51(m,1H),5.14-4.91(m，2H)，4.77-4.54(m,3H),4.27-3.83(m，4H),3.81-3.39(m,4H),3.01-2.56(m，4H)，2.47(s，3H)，2.45-2.27(m,3H),2.26-2.17(m,1H),2.18(s,3H)。
Example 8: synthesis of Compound 2
Figure BDA0003225850520000261
Compound 2 is prepared by the following method
Figure BDA0003225850520000262
Synthetic solid of compound 2: n is a radical of hydrogen 2 In a three-necked flask with an internal temperature thermometer, 2-fluoroacrylic acid (9.4 mg,0.10 mmol), 2- (7-azobenzotriazol) -N, N, N ', N' -tetramethyluronium hexafluorophosphate (39.9mg, 0.10 mmol), compounds 1 to 2 (45.0 mg, 0.07mmol), and N, N-diisopropylethylamine (27.0 mg, 0.21mmol) were added to N, N-dimethylformamide (1.0 ml) under protection, and the reaction was stirred at 25 ℃ for 1 hour and monitored by LC-MS for completion. The reaction solution was filtered, and the filtrate was collected and purified by Prep-HPLC (acetonitrile/TFA aq (0.01%)) to obtain Compound 2 as an off-white solid (9.3 mg, yield: 19%, purity: 99%).
MS(ESI):m/z＝677.2[M+H] + ；
1 H NMR(400MHz,Methanol-d4):δ6.75(s,1H),5.68-5.52(m,1H),5.43-5.27(m，2H)，5.12-4.95(m，2H)，4.77-4.60(m，3H)，4.12-3.85(m,4H),3.74-3.62(m,2H),3.55-3.41(m,2H),3.06-2.88(m,2H)，2.82-2.58(m,2H),2.49(s,3H),2.56-2.25(m,4H),2.20(s，3H)。
The example compounds in table one can be further obtained using a synthetic route analogous to that of compound 1, using commercial starting materials or known intermediates.
Watch 1
Figure BDA0003225850520000271
Figure BDA0003225850520000281
Figure BDA0003225850520000291
Figure BDA0003225850520000301
Example 26: inhibitory Activity of partial Compounds of the invention on SOS 1-mediated conversion of KRas (G12C) -GDP to KRas (G12C) -GTP
The inhibitory activity of some of the compounds of the invention on SOS 1-mediated conversion of KRas (G12C) -GDP to KRas (G12C) -GTP was determined by the following method
The experimental steps are as follows:
1) KRAS (G12C) and GDP at low Mg concentrations 2+ Pre-incubation in buffer for 30min, using high concentration Mg 2+ The reaction was terminated.
2) After the reaction, the solution was desalted by a column to obtain KRAS (G12C) -GDP, and the KRAS (G12C) -GDP protein concentration was measured by Brandford method.
3) Test compounds were diluted in DMSO at 3-fold gradients, 11 concentration gradients. Gradient dilutions of 50nL of compound were transferred to 384 reaction plates using Echo, 2 replicates per concentration.
4) To the 384 reaction plates containing the compound, 2.5. Mu.L of KRAS (G12C) -GDP solution per well was added, centrifuged at 1000rpm for 1min, and incubated at 25 ℃ for 15min.
5) Add 2.5. Mu.L of the SOS1 and GTP mixture, centrifuge at 1000rpm for 1min, and incubate at 25 ℃ for 2h.
6) Add 5. Mu.L of cRAF RBD solution, centrifuge at 1000rpm for 1min, incubate at 25 ℃ for 1h.
7) Add 10. Mu.L Ab Anti-6HIS Tb cryptate Gold donor and MAb Anti GST-XL665 receptor mixture, centrifuge at 1000rpm for 1min, incubate at 25 ℃ for 1h.
8) After the incubation, the ratio of 665/615nm was read using an Envision multifunctional plate reader.
9) IC of test compounds was calculated and analyzed using a four parameter logistic regression model (4 PL) with GraphPad Prism software 50 And (4) horizontal.
The experimental data processing method comprises the following steps:
the IC of the compound was obtained using the following non-linear fit equation 50 (median inhibitory concentration):
Y＝Bottom+(Top-Bottom)/(1+10^((LogIC50-X)*HillSlope))
x log value of Compound concentration
Y:Ratio(665nm/615nm)
The experimental results are shown in the second table:
a second table: inhibitory Activity of partial Compounds of the invention on SOS 1-mediated conversion of KRas (G12C) -GDP to KRas (G12C) -GTP
Figure BDA0003225850520000311
Figure BDA0003225850520000321
As can be seen from Table two, the compounds of the present invention showed better inhibitory activity for inhibiting the conversion of KRas (G12C) -GDP to KRas (G12C) -GTP mediated by SOS1, and some of the compounds showed comparable or better inhibitory activity than the control drug AMG-510. This indicates that the compounds of the invention are expected to be useful in the treatment of diseases caused/associated with KRas G12C mutations.
It will be apparent to those skilled in the art that various changes and modifications may be made in the present invention without departing from the spirit and scope of the invention. Thus, if such modifications and variations of the present invention fall within the scope of the claims of the present invention and their equivalents, the present invention is also intended to include such modifications and variations.

Claims (17)

1. A heterocyclic compound having the formula (I) or a stereoisomer, pharmaceutically acceptable salt thereof:
Figure FDA0003225850510000011
wherein the content of the first and second substances,
R 1 is C optionally substituted by 1 to 4 substituents 6-10 Aryl or 5-10 membered heteroaryl, said substituents being selected from: halogen, hydroxy, mercapto, amino, nitro, cyano, C 1-6 Alkyl radical, C 3-6 Cycloalkyl radical, C 1-6 Alkylamino radical, di (C) 1-6 Alkyl) amino, halo C 1-6 Alkyl, hydroxy C 1-6 Alkyl, ammoniaRadical C 1-6 Alkyl radical, C 1-6 Alkoxy radical, C 1-6 Alkylthio, halo C 1-6 Alkoxy, halo C 1-6 Alkylthio, hydroxy C 1-6 Alkoxy, hydroxy C 1-6 Alkylthio, amino C 1-6 Alkoxy or amino C 1-6 An alkylthio group;
R 2 is halogen, hydroxy, mercapto, amino, nitro, cyano, C 1-6 Alkyl radical, C 3-6 Cycloalkyl radical, C 1-6 Alkylamino radical, di (C) 1-6 Alkyl) amino, C 1-6 A haloalkyl group;
l is selected from-O-, -S-, -CONH-, -NH-;
y is N or CH;
m is 0, 1,2 or 3;
R 3 is selected from optionally substituted 1-3R 3a Substituted 3-10 membered cycloalkyl or 3-10 membered heterocyclyl;
each R 3a Independently selected from: halogen, hydroxy, mercapto, amino, nitro, cyano, C 1-6 Alkyl radical, C 3-6 Cycloalkyl, C 1-6 Alkylamino radical, di (C) 1-6 Alkyl) amino, di (C) 1-6 Alkyl) amino-C 1-3 Alkyl radical, C 1-6 Haloalkyl, hydroxy C 1-6 Alkyl, amino C 1-6 Alkyl radical, C 1-6 Alkoxy radical, C 1-6 Alkylthio, halogeno C 1-6 Alkoxy radical, C 1-6 Haloalkylthio, hydroxy C 1-6 Alkoxy, hydroxy C 1-6 Alkylthio, amino C 1-6 Alkoxy or C 1-6 An aminoalkylthio group;
X 1 and X 2 Are each independently CH or N;
n is 0, 1,2 or 3;
each R is 4 Independently represent halogen, hydroxyl, sulfydryl, amino, nitro, cyano C 1-6 Alkyl radical, C 1-6 Alkyl radical, C 1-6 Alkylamino radical, di (C) 1-6 Alkyl) amino, halo C 1-6 Alkyl, hydroxy C 1-6 Alkyl, amino C 1-6 Alkyl, mercapto C 1-6 Alkyl radical, C 1-6 Alkoxy radical C 1-6 Alkyl or halo C 1-6 Alkoxy radical C 1-6 An alkyl group;
R 5 is-CO-CH = CHR 5a or-CO-CR 5a ＝CH 2 ，R 5a Each independently selected from hydrogen, amino, cyano, halogen, -CO-NH-C 1-6 Alkyl, -CO-N (C) 1-6 Alkyl radical) 2 、C 1-6 Alkyl radical, C 1-6 Alkoxy, halo C 1-6 Alkoxy radical, C 1-6 Alkylamino radical, di (C) 1-6 Alkyl) amino, halo C 1-6 Alkyl, hydroxy C 1-6 Alkyl or amino C 1-6 An alkyl group.
2. A compound according to claim 1, or a stereoisomer, pharmaceutically acceptable salt thereof: r is 1 Is C optionally substituted with 1 to 4 substituents 6-10 Aryl or 5-10 membered heteroaryl, said substituents being selected from: halogen, hydroxy, mercapto, amino, nitro, cyano, C 1-6 Alkyl radical, C 1-6 Alkylamino radical, di (C) 1-6 Alkyl) amino, halo C 1-6 Alkyl, hydroxy C 1-6 Alkyl radical, C 1-6 Alkoxy, halo C 1-6 An alkoxy group.
3. A compound according to claim 2, or a stereoisomer, pharmaceutically acceptable salt thereof:
R 1 is a group optionally substituted with 1 to 4 substituents:
Figure FDA0003225850510000021
Figure FDA0003225850510000022
the substituent is selected from: halogen, hydroxy, amino, C 1-6 Alkyl, halo C 1-6 An alkyl group.
4. A compound according to claim 3, or a stereoisomer, pharmaceutically acceptable salt thereof,
R 1 is the following fragment:
Figure FDA0003225850510000023
5. a compound according to claim 1, or a stereoisomer, pharmaceutically acceptable salt thereof, wherein R 2 Methyl, ethyl, propyl, isopropyl.
6. A compound according to claim 1, or a stereoisomer, pharmaceutically acceptable salt thereof, wherein L is from-O-, -S-.
7. The compound according to claim 1, or a stereoisomer, pharmaceutically acceptable salt thereof, Y is N.
8. A compound according to claim 1, or a stereoisomer, pharmaceutically acceptable salt thereof, wherein m is 1.
9. A compound according to claim 1 or a stereoisomer, pharmaceutically acceptable salt thereof, R 3 Is selected from optionally substituted by 1-3R 3a Substituted: cyclopropyl, cyclopentyl, cyclohexyl, tetrahydrofuryl, tetrahydrothienyl, tetrahydropyrrolyl, tetrahydroimidazoyl, tetrahydropyrazolyl, tetrahydrothiazolyl, tetrahydrooxazolyl, tetrahydropyranyl, piperidinyl, piperazinyl, morpholinyl, or hexahydropyrimidyl/cis-3-azabicyclo [3,3,0 ] group]Octane;
each R 3a Independently selected from: halogen, hydroxy, amino, cyano, C 1-6 Alkyl, N dimethylaminomethyl, C 1-6 A haloalkyl group.
10. A compound according to claim 9 or a stereoisomer, pharmaceutically acceptable salt thereof, R 3 The following fragments:
Figure FDA0003225850510000031
11. a compound according to claim 1 or a stereoisomer, pharmaceutically acceptable salt thereof, X 1 And X 2 Are all N.
12. A compound according to claim 1, or a stereoisomer, pharmaceutically acceptable salt thereof, n is 0 or 1, each R 4 Independently of each other, methyl and acetonitrile.
13. A compound according to claim 1 or a stereoisomer, pharmaceutically acceptable salt thereof, R 5 is-CO-CR 5 ＝CH 2 ，R 5 Selected from hydrogen and fluorine.
14. The compound according to claim 1, or a stereoisomer, pharmaceutically acceptable salt thereof, having the following specific compound,
Figure FDA0003225850510000041
15. a compound according to claims 1-14, or a stereoisomer, pharmaceutically acceptable salt thereof, for use in the manufacture of a medicament for the treatment and/or prevention of cancer.
16. The use according to claim 15, wherein the cancer is mediated by Kras mutation.
17. The use according to claim 15, wherein said cancer is selected from lung cancer, colorectal cancer or pancreatic cancer.
CN202110971266.0A 2021-08-24 2021-08-24 Heterocyclic compound with KRAS (KRAS-mutated protein) inhibition effect and pharmaceutical composition and application thereof Pending CN115716840A (en)

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