NMT inhibitor, and preparation method therefor and use thereof

Abstract

Provided in the present invention are an NMT inhibitor, and a preparation method therefor and the use thereof. Specifically, the present invention relates to a compound as represented by formula (1) and a preparation method therefor, and a pharmaceutical composition containing the compound as represented by formula (1) and / or a pharmaceutically acceptable salt thereof, and a preparation method therefor and the use thereof as an NMT inhibitor in the preparation of an anti-tumor drug.

Description

NMT inhibitors and their preparation and use Technical Field

[0001] The present invention relates to the field of medicinal chemistry, and more specifically, to a class of NMT inhibitors having a structure as shown in formula (1), a preparation method thereof, and the use of such compounds in preparing pharmaceutical compositions for treating, regulating and / or preventing diseases mediated by NMT. Background Art

[0002] NMT (N-myristoyltransferase, NMT, N-myristoyltransferase) is an enzyme that catalyzes the myristoylation of more than 100 proteins in human cells and affects downstream related signaling pathways. In humans, protein myristoylation is mediated by two ubiquitously expressed N-myristoyltransferases, NMT1 and NMT2. Myristoylation is a co-translational and post-translational modification reaction in eukaryotes. Through the action of NMT1 and NMT2, the myristoyl group is transferred to the N-terminal glycine of the substrate protein [Cell Death and Disease, 2018, 9(12): 1143]. It plays a role similar to a "switch" and can trigger a variety of reversible protein-cell membrane and protein-protein interactions.

[0003] NMT1 is highly expressed in a variety of tumor tissues. In liver cancer, NMT1 is associated with poor prognosis. Compared with patients with low expression, patients with high NMT1 expression have shorter survival. The expression of NMT1 has important clinical significance. The primary breast cancer tissue and adjacent tissue of 20 breast cancer patients were detected. The results showed that compared with adjacent tissue, the expression of NMT1 in breast cancer tissue was significantly increased, especially in triple-negative breast cancer tissue [Cell Death and Disease, 2018, 9 (12): 1143]. Elevated NMT1 is associated with tumor development and shorter survival of patients. In patients with colorectal cancer, NMT1 expression is elevated in peripheral blood and bone marrow and can be used as a diagnostic marker for colorectal cancer [J Transl Med. 2007, 5: 58]. High expression of NMT1 is associated with poor prognosis of ovarian cancer and the progression of liver cancer, brain cancer, etc. [Annals of medicine, 2023, 55 (1): 1422-1430].

[0004] NMT1 catalyzes its substrate proteins to form myristoylated proteins, activates downstream signaling pathways such as NF-KB, C-Myc, and ERK, and promotes tumor cell proliferation, migration, and metastasis. Inhibition of NMT1 can induce cell cycle arrest, inhibit cell proliferation and malignant growth [Annals of medicine, 2023, 55(1): 1422-1430]. NMT1 is highly expressed in a variety of tumors and is associated with tumor staging, prognosis, and survival. It is a potential target for precision tumor therapy. NMT inhibitors can significantly inhibit the activity of blood tumor cells, including B lymphocytes, inhibit the myristoylation of lymphoma cells, and inhibit the signal transduction of B cell receptors, leading to cell death and inhibiting tumor growth in mouse tumor models [Nature communications, 2020, 11(1): 5348].

[0005] Although some NMT small molecule inhibitors have been disclosed, there is still a need to develop new compounds with better activity and pharmacokinetic properties. The NMT inhibitor compound of the present invention having the structure represented by general formula (1) exhibits excellent effects and functions, and can provide a new option for the precision treatment of tumors.

[0006] Summary of the Invention

[0007] The purpose of the present invention is to provide a novel NMT inhibitor with high activity and its preparation method and use.

[0008] In the first aspect of the present invention, there is provided a compound having a structure as shown in formula (1), an optical isomer thereof or a pharmaceutically acceptable salt thereof:

[0009] In formula (1):

[0010] n is 0, 1, 2, or 3;

[0011] X 1 and X 2 Each independently selected from the group consisting of CH, CR 1 or N;

[0012] W is selected from the group consisting of a chemical bond, O, S, or NH;

[0013] Y is selected from the group consisting of: chemical bonds, -(CH2) m -、-(CH2) m O- or -(CH2) m O-(CH2) m , wherein m is selected from 1, 2 or 3;

[0014] L is -(CH2) m -;

[0015] Each R1 Each independently selected from the group consisting of D, hydroxy, halogen, CN, NO2, NH2, C 1-4 Alkyl, C 2-4 Alkenyl, C 2-4 Alkynyl, C 1-4 Halogenated alkyl, C 1-4 Alkoxy, C 1-4 Halogenated alkoxy, C 3-6 alkyl saturated or partially unsaturated carbocyclic group, 3-8 membered saturated or partially unsaturated heterocyclic group, C 6-10 Aryl or 5-12 membered heteroaryl;

[0016] R 2 -NR 6 R 7 , where R 6 and R 7 Each independently selected from the following group: H, D, C 1-6 Alkyl, C 1-6 alkyl halide;

[0017] Or, R 6 and R 7 and the nitrogen atom to which it is connected together form a 3-8 membered saturated or partially unsaturated heterocyclic group, or a 5-12 membered heteroaryl group;

[0018] R 3 Selected from the following groups: H, D, C 1-4 Alkyl, C 1-4 Halogenated alkyl, C 1-4 Alkoxy, C 1-4 Halogenated alkoxy, C 3-6 alkyl saturated or partially unsaturated carbocyclic group, 3-8 membered saturated or partially unsaturated heterocyclic group, C 6-10 Aryl or 5-12 membered heteroaryl;

[0019] R 4 Select from the following group: C 1-4 Alkyl, C 1-4 Halogenated alkyl, C 3-6 alkyl saturated or partially unsaturated carbocyclic group, 3-8 membered saturated or partially unsaturated heterocyclic group, C 6-10 Aryl or 5-12 membered heteroaryl;

[0020] R 5 Selected from the group consisting of hydroxyl, CN, C 1-6 Alkyl, C 1-6 Halogenated alkyl, hydroxy substituted C 1-6 Alkyl, cyano substituted C 1-6 Alkyl or -C(O)NR 8 R 9 ; R 8 and R 9Each independently selected from the following group: H, D, C 1-6 Alkyl, C 1-6 alkyl halide;

[0021] Or, R 8 and R 9 and the nitrogen atom to which it is connected together form a 3-8 membered saturated or partially unsaturated heterocyclic group, or a 5-12 membered heteroaryl group;

[0022] wherein the alkyl, alkenyl, alkynyl, haloalkyl, alkoxy, haloalkoxy, carbocyclic, heterocyclic, aryl, and heteroaryl groups are each independently and optionally substituted with one or more substituents selected from the group consisting of D, hydroxyl, halogen, CN, NO2, NH2, C 1-4 Alkyl, C 2-4 Alkenyl, C 2-4 Alkynyl, C 1-4 Halogenated alkyl, C 1-4 Alkoxy or C 1-4 Halogenated alkoxy.

[0023] In another preferred embodiment, wherein in the formula (1), each R 1 Each independently selected from the group consisting of F, Cl, Br, CN, NO2, C 1-3 Alkyl, C 1-3 Halogenated alkyl, C 1-3 Alkoxy or C 1-3 Halogenated alkoxy.

[0024] In another preferred embodiment, wherein in the formula (1), R 2 -NR 6 R 7 , where R 6 and R 7 Each independently selected from the following group: H, D, C 1-3 Alkyl, C 1-3 Halogenated alkyl.

[0025] In another preferred embodiment, wherein in the formula (1), R 3 Selected from the following groups: H, D, C 1-3 Alkyl, C 1-3 Halogenated alkyl.

[0026] In another preferred embodiment, wherein in the formula (1), R 4 Select from the following group: C 1-3 Alkyl, C 1-3 Halogenated alkyl.

[0027] In another preferred embodiment, wherein in the formula (1), R 5 Select from the following groups:

[0028] CN、C1-6 Alkyl, C 1-6 Halogenated alkyl, hydroxy substituted C 1-6 Alkyl, cyano substituted C 1-6 Alkyl or -C(O)NR 8 R 9 ;

[0029] Among them, R 8 and R 9 Each independently selected from the following group: H, D, C 1-3 Alkyl, C 1-3 Halogenated alkyl.

[0030] In another preferred embodiment, the representative compound of the present invention has a structure selected from the following group:

[0031] In a second aspect of the present invention, a pharmaceutical composition for treating, regulating and / or preventing diseases mediated by NMT is provided, the pharmaceutical composition comprising:

[0032] (1) a compound according to the first aspect of the present invention, or an isomer, crystal form, pharmaceutically acceptable salt, hydrate or solvate thereof as an active ingredient; and

[0033] Optional (2) a pharmaceutically acceptable excipient or carrier.

[0034] In the third aspect of the present invention, there is provided a use of the compound according to the first aspect of the present invention, its isomer, crystal form, pharmaceutically acceptable salt, hydrate or solvate, or the pharmaceutical composition according to the second aspect of the present invention for preparing a pharmaceutical composition for treating, regulating and / or preventing diseases mediated by NMT.

[0035] In another preferred embodiment, the NMT-mediated related diseases include infectious diseases or hyperproliferative diseases.

[0036] In another preferred embodiment, the infectious disease includes protozoan infection, such as malaria and leishmaniasis.

[0037] In another preferred embodiment, the infectious disease includes viral infection, such as human rhinovirus and HIV.

[0038] In another preferred embodiment, the hyperproliferative disease is selected from the group consisting of lymphoma, leukemia, brain tumor, gastric cancer, liver cancer, lung cancer, intestinal cancer, pancreatic cancer, breast cancer, cervical cancer, endometrial cancer and prostate cancer.

[0039] It should be understood that within the scope of the present invention, the above-mentioned technical features of the present invention and the technical features described in detail below (such as in the embodiments) can be combined with each other to form new or preferred technical solutions. Due to space limitations, they will not be listed here one by one. DETAILED DESCRIPTION

[0040] After extensive and in-depth research, extensive screening, and testing, the present inventors have discovered for the first time a class of compounds represented by formula (1) that have significant therapeutic effects on diseases mediated by NMT. ​​The compounds of the present invention have excellent inhibitory activity against the NMT1 enzyme and also exhibit good cell proliferation inhibition activity against the MV-4-11 cell line. This invention was completed on this basis.

[0041] Compounds of the present invention and their synthesis

[0042] The present invention provides an inhibitor targeting NMT, namely a compound of formula (1), or its isomers, crystal forms, pharmaceutically acceptable salts (inorganic or organic salts), hydrates or solvates. Preferably, the compound of the present invention is as described in the first aspect.

[0043] The present invention also provides a method for preparing the compound of formula (1). The following specifically describes the method for preparing the compound of general formula (1), but these specific methods do not constitute any limitation to the present invention.

[0044] In one aspect, the compounds described herein are prepared according to methods known in the art. However, the conditions of the methods, such as reactants, solvents, bases, amounts of the compounds used, reaction temperatures, reaction times, etc., are not limited to the following explanations. The compounds of the present invention can also be conveniently prepared by optionally combining various synthetic methods described in this specification or known in the art, and such combinations can be easily performed by those skilled in the art. In one aspect, the present invention also provides a method for preparing the compound represented by the general formula (1), which is prepared using the following general reaction scheme 1:

[0045] General reaction scheme 1:

[0046] The embodiment of the compound of formula (1) can be prepared according to the general reaction scheme 1, wherein Z is halogen, Q is boronic acid or boronic ester, M is halogen, OMs, OTos, etc., and R 1 、R 2 、R 3 、R 4 、R 5 、X 1 、X 2 , L, W, Y and n are as defined above.

[0047] Related definitions

[0048] Unless otherwise indicated, the following terms and phrases used herein are intended to have the following meanings. A particular term or phrase should not be construed as indefinite or unclear unless specifically defined, but rather should be understood in accordance with its ordinary meaning. When a trade name appears in this document, it is intended to refer to the corresponding commercial product or its active ingredient.

[0049] The term "pharmaceutically acceptable" as used herein refers to those compounds, materials, compositions and / or dosage forms that are, within the scope of sound medical judgment, suitable for use in contact with the tissues of human beings and animals without excessive toxicity, irritation, allergic response or other problems or complications, commensurate with a reasonable benefit / risk ratio.

[0050] The term "pharmaceutically acceptable salt" refers to salts of the compounds of the present invention, prepared by reacting the compounds of the present invention with relatively nontoxic acids or bases. When the compounds of the present invention contain relatively acidic functional groups, base addition salts can be obtained by contacting the compounds with a sufficient amount of base in neat solution or in a suitable inert solvent. Pharmaceutically acceptable base addition salts include sodium, potassium, calcium, ammonium, organic amine, or magnesium salts, or similar salts. When the compounds of the present invention contain relatively basic functional groups, acid addition salts can be obtained by contacting the compounds with a sufficient amount of acid in neat solution or in a suitable inert solvent. Examples of pharmaceutically acceptable acid addition salts include inorganic acid salts such as hydrochloric acid, hydrobromic acid, nitric acid, carbonic acid, bicarbonate, phosphoric acid, monohydrogen phosphate, dihydrogen phosphate, sulfuric acid, bisulfate, hydroiodic acid, phosphorous acid, and the like; and organic acid salts such as acetic acid, propionic acid, isobutyric acid, maleic acid, malonic acid, benzoic acid, succinic acid, suberic acid, fumaric acid, lactic acid, mandelic acid, phthalic acid, benzenesulfonic acid, p-toluenesulfonic acid, citric acid, tartaric acid, and methanesulfonic acid; and salts of amino acids (such as arginine) and organic acids such as glucuronic acid. Certain specific compounds of the present invention contain both basic and acidic functional groups and can be converted into either base or acid addition salts.

[0051] Pharmaceutically acceptable salts of the present invention can be synthesized by conventional chemical methods from parent compounds containing acid radicals or bases. Generally, such salts are prepared by reacting these compounds in free acid or base form with a stoichiometric amount of a suitable base or acid in water or an organic solvent or a mixture of the two.

[0052] 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, and racemic mixtures 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. All such isomers and mixtures thereof are included within the scope of the present invention.

[0053] Unless otherwise indicated, the term "enantiomer" or "optical isomer" refers to stereoisomers that are mirror images of one another.

[0054] Unless otherwise indicated, the term "cis-trans isomers" or "geometric isomers" arises from the inability to rotate freely about double bonds or single bonds forming ring carbon atoms.

[0055] Unless otherwise indicated, the term "diastereomer" refers to stereoisomers that have two or more chiral centers and that are not mirror images of each other.

[0056] Unless otherwise indicated, "(D)" or "(+)" indicates dextrorotatory, "(L)" or "(-)" indicates levorotatory, and "(DL)" or "(±)" indicates racemic.

[0057] Unless otherwise specified, use a solid wedge key. and dotted wedge key To indicate the absolute configuration of a stereocenter, use a straight solid bond and straight dashed key Indicate the relative configuration of stereocenters with a wavy line Indicates a wedge-shaped solid key or dotted wedge key Or use a wavy line Indicates a straight solid bond and straight dashed key

[0058] Unless otherwise indicated, the term "isomer excess" or "enantiomeric excess" refers to the difference between the relative percentages of two isomers or two enantiomers. For example, if the content of one isomer or enantiomer is 90% and the content of the other isomer or enantiomer is 10%, the isomer or enantiomeric excess (ee value) is 80%.

[0059] Optically active (R)- and (S)-isomers, as well as D and L isomers, can be prepared by chiral synthesis or chiral reagents or other conventional techniques. If one enantiomer of a compound of the present invention is desired, it can be prepared by asymmetric synthesis or derivatization with a chiral auxiliary, wherein the resulting diastereomeric mixture is separated and the auxiliary group is cleaved to provide the pure desired enantiomer. Alternatively, when the molecule contains a basic functional group (such as an amino group) or an acidic functional group (such as a carboxyl group), a diastereomeric salt is formed with an appropriate optically active acid or base, and then the diastereoisomers are separated by conventional methods known in the art, and then the pure enantiomer is recovered. In addition, the separation of enantiomers and diastereomers is usually accomplished by using chromatography, which employs a chiral stationary phase and is optionally combined with a chemical derivatization method (e.g., carbamate formation from an amine).

[0060] The compounds of the present invention may contain unnatural proportions of atomic isotopes at one or more of the atoms that constitute the compound. For example, the compounds may be labeled with radioactive isotopes, such as tritium ( 3 H), iodine-125 ( 125 I) or C-14( 14 C). For example, deuterated compounds can be formed by replacing hydrogen atoms with heavy hydrogen. The bond formed by deuterium and carbon is stronger than the bond formed by ordinary hydrogen and carbon. Compared with non-deuterated drugs, deuterated drugs generally have advantages such as reduced toxic side effects, increased drug stability, enhanced efficacy, and prolonged drug half-life in vivo. All isotopic variations of the compounds of the present invention, whether radioactive or not, are included within the scope of this invention.

[0061] "Optional" or "optionally" means that the subsequently described event or circumstance may but need not occur, and that the description includes instances where said event or circumstance occurs and instances where it does not.

[0062] The term "substituted" means that any one or more hydrogen atoms on a particular atom are replaced by a substituent, which may include deuterium and hydrogen variants, as long as the valence state of the particular atom is normal and the substituted compound is stable. When the substituent is oxygen (i.e., =O), it means that two hydrogen atoms are replaced. Oxygen substitution does not occur on aromatic groups. The term "optionally substituted" means that it may be substituted or unsubstituted, and unless otherwise specified, the type and number of substituents can be any on the basis of chemical achievable.

[0063] When any variable (e.g., R) occurs more than once in a compound's composition or structure, its definition at each occurrence is independent. Thus, for example, if a group is substituted with 0-2 Rs, the group may be optionally substituted with up to two Rs, with each occurrence of R being an independent choice. Furthermore, combinations of substituents and / or their variants are permissible only if such combinations result in stable compounds.

[0064] When the number of a linking group is 0, such as -(CH2)0-, it means that the linking group is a single bond.

[0065] When one of the variables is selected from a chemical bond, it means that the two groups it connects are directly connected. For example, when L in XLY represents a chemical bond, it means that the structure is actually XY.

[0066] Unless otherwise specified, C n-n+m or C n -C n+m Any specific case including n to n+m carbons, such as C 1-12 Including C1, C2, C3, C4, C5, C6, C7, C8, C9, C 10 、C 11 and C 12 , also includes any range from n to n+m, such as C 1-12 Including C 1-3 、C 1-6 、C 1-9 、C 3-6 、C 3-9 、C 3-12 、C 6-9 、C 6-12 and C 9-12 Similarly, n-membered to n+m-membered means that the number of atoms in the ring is n to n+m, for example, a 3-12-membered ring includes a 3-membered ring, a 4-membered ring, a 5-membered ring, a 6-membered ring, a 7-membered ring, an 8-membered ring, a 9-membered ring, a 10-membered ring, an 11-membered ring and a 12-membered ring, and also includes any range from n to n+m, for example, a 3-12-membered ring includes a 3-6-membered ring, a 3-9-membered ring, a 5-6-membered ring, a 5-7-membered ring, a 6-7-membered ring, a 6-8-membered ring and a 6-10-membered ring, etc.

[0067] Unless otherwise specified, “C 1-6 "Alkyl" is used to represent a straight or branched saturated aliphatic hydrocarbon group consisting of 1 to 6 carbon atoms. 1-6 Alkyl groups include C 1-2 、C 1-3 、C 1-4 、C 1-5 、C 2-4 、C 2-6 、C 3-5, C5 and C6 alkyl, etc.; which can be monovalent (such as methyl), divalent (such as methylene) or polyvalent (such as methine). Non-limiting examples of C1-6 alkyl include methyl, ethyl, propyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, sec-butyl, n-pentyl, 1,1-dimethylpropyl, 1,2-dimethylpropyl, 2,2-dimethylpropyl, 1-ethylpropyl, 2-methylbutyl, 3-methylbutyl, n-hexyl, 1-ethyl-2-methylpropyl, 1,1,2-trimethylpropyl, 1,1-dimethylbutyl, 1,2-dimethylbutyl, 2,2-dimethylbutyl, 1,3-dimethylbutyl, 2,3-dimethylbutyl, 2-ethylbutyl, 2-methylpentyl, 3-methylpentyl, 4-methylpentyl, and various branched chain isomers thereof.

[0068] Unless otherwise specified, “C 1-3 "Alkyl" is used to represent a straight or branched saturated aliphatic hydrocarbon group consisting of 1 to 3 carbon atoms. 1-3 Alkyl groups include C 1-2 and C 2-3 Alkyl, etc.; it can be monovalent (such as methyl), divalent (such as methylene) or polyvalent (such as methine). 1-3 Non-limiting examples of alkyl groups include methyl, ethyl, propyl, n-propyl, isopropyl, and the like.

[0069] Unless otherwise specified, “C 1-3 "Alkoxy" refers to an alkyl group containing 1 to 3 carbon atoms attached to the rest of the molecule through an oxygen atom. 1-3 Alkoxy groups include C 1-2 , C2 and C3 alkoxy, etc.; C 1-3 Non-limiting examples of alkoxy groups include methoxy, ethoxy, propoxy, n-propoxy, isopropoxy, and the like.

[0070] "Halogen" or "halo" refers to fluorine, chlorine, bromine or iodine.

[0071] "Hydroxyl" refers to -OH.

[0072] "Cyano" refers to -CN.

[0073] Specific pharmaceutical and medical terms

[0074] The term "acceptable," as used herein, means that a prescribed ingredient or active ingredient has no undue adverse effect on health and well-being for the general purpose of treatment.

[0075] The terms "treat," "treatment," or "therapy" as used herein include alleviating, inhibiting, or ameliorating the symptoms of a disease or condition; inhibiting the development of complications; ameliorating or preventing underlying metabolic syndrome; inhibiting the development of a disease or symptom, such as controlling the progression of a disease or condition; alleviating a disease or symptom; causing a regression of a disease or symptom; alleviating complications caused by a disease or symptom, or preventing or treating signs caused by a disease or symptom. As used herein, a compound or pharmaceutical composition, upon administration, can improve a disease, symptom, or condition, particularly by improving its severity, delaying its onset, slowing its progression, or reducing its duration. Whether the administration is fixed or temporary, continuous or intermittent, the circumstances attributable to or related to the administration can be explained.

[0076] "Active ingredient" refers to the compound shown in the general formula (1), as well as the pharmaceutically acceptable inorganic or organic salts of the compound of formula (1). The compounds of the present invention may contain one or more asymmetric centers (axial chirality) and therefore appear in the form of racemates, racemic mixtures, single enantiomers, diastereomeric compounds and single diastereomers. The asymmetric centers that may exist depend on the properties 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 as well as pure or partially pure compounds are included within the scope of the present invention. The present invention is meant to include all such isomeric forms of these compounds.

[0077] The terms "compound," "composition," "agent," or "medicine or medicament" are used interchangeably herein and refer to a compound or composition that, when administered to a subject (human or animal), induces a desired pharmaceutical and / or physiological response through local and / or systemic action.

[0078] The term "administered," "administering," or "administration" as used herein refers to the direct administration of the compound or composition, or the administration of a prodrug, derivative, or analog of the active compound.

[0079] Although the numerical ranges and parameters used to define the broader scope of the present invention are approximate, the numerical values ​​of the specific examples are presented herein as precisely as possible. However, any numerical value inherently and inevitably contains standard deviations resulting from individual testing methods. As used herein, "about" generally refers to the actual value being 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 an acceptable standard error of the mean, as determined by one skilled in the art. Except in the experimental examples, or unless otherwise expressly indicated, all ranges, amounts, values, and percentages used herein (e.g., to describe material amounts, time periods, temperatures, operating conditions, quantitative ratios, and the like) are to be understood as modified by the word "about." Therefore, unless otherwise indicated, the numerical parameters disclosed in this specification and the appended claims are approximate and may be modified as needed. At a minimum, these numerical parameters should be understood to include the number of significant digits indicated and to include normal rounding.

[0080] Unless otherwise defined in this specification, the scientific and technical terms used herein have the same meanings as commonly understood by those skilled in the art. In addition, unless otherwise defined in this specification, singular terms used in this specification include the plural form of the term, and plural terms also include the singular form of the term, unless otherwise defined in the context.

[0081] Route of administration

[0082] The compounds of the present invention and their pharmaceutically acceptable salts can be formulated into various formulations containing a safe and effective amount of the compounds of the present invention or their pharmaceutically acceptable salts and a pharmacologically acceptable excipient or carrier. "Safe and effective amount" means an amount of the compound sufficient to significantly improve the condition without causing serious side effects. The safe and effective amount of the compound will be determined based on the patient's age, condition, and duration of treatment, among other factors.

[0083] "Pharmaceutically acceptable excipients or carriers" refer to: one or more compatible solid or liquid fillers or gel substances, which are suitable for human use and must have sufficient purity and sufficiently low toxicity. "Compatibility" here means that the components in the composition can be mixed with the compounds of the present invention and with each other without significantly reducing the efficacy of the compounds. Some examples of pharmacologically acceptable excipients or carriers include cellulose and its derivatives (such as sodium carboxymethyl cellulose, sodium ethyl cellulose, cellulose acetate, etc.), gelatin, talc, solid lubricants (such as stearic acid, magnesium stearate), calcium sulfate, vegetable oils (such as soybean oil, sesame oil, peanut oil, olive oil, etc.), polyols (such as propylene glycol, glycerol, mannitol, sorbitol, etc.), emulsifiers (such as ), wetting agents (such as sodium lauryl sulfate), colorants, flavorings, stabilizers, antioxidants, preservatives, pyrogen-free water, etc.

[0084] The compounds of the present invention may be administered orally, rectally, parenterally (intravenously, intramuscularly or subcutaneously), or topically.

[0085] 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 solubilizers, for example, starches, lactose, sucrose, glucose, mannitol, and silicic acid; (b) binders, for example, hydroxymethylcellulose, alginates, gelatin, polyvinyl pyrrolidone, sucrose, and acacia; (c) humectants, for example, glycerol; (d) disintegrants, for example, agar, calcium carbonate, potato or tapioca starch, alginic acid, certain complex silicates, and sodium carbonate; (e) solubilizers, for example, paraffin; (f) absorption accelerators, for example, quaternary ammonium compounds; (g) wetting agents, for example, cetyl alcohol and glyceryl 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 the case of capsules, tablets and pills, the dosage forms may also comprise buffering agents.

[0086] Solid dosage forms such as tablets, dragees, capsules, pills, and granules can be prepared using coatings and shell materials, such as enteric coatings and other materials known in the art. They may contain opacifying agents, and the release of the active compound or compounds in such compositions can be delayed in a certain portion of the digestive tract. Examples of useful encapsulating components are polymeric substances and waxes. If desired, the active compound can also be microencapsulated with one or more of the above-mentioned excipients.

[0087] Liquid dosage forms for oral administration include pharmaceutically acceptable emulsions, solutions, suspensions, syrups or tinctures. In addition to the active compound, the liquid dosage form may contain an inert diluent conventionally used in the art, such as water or other solvents, solubilizers and emulsifiers, for example, ethanol, isopropyl alcohol, ethyl carbonate, ethyl acetate, propylene glycol, 1,3-butylene glycol, dimethylformamide, and oils, particularly cottonseed oil, peanut oil, corn germ oil, olive oil, castor oil and sesame oil, or mixtures thereof.

[0088] Besides such inert diluents, the composition may also include adjuvants such as wetting agents, emulsifying and suspending agents, sweetening, flavoring, and perfuming agents.

[0089] Suspensions, in addition to the active compounds, may contain suspending agents such as, for example, ethoxylated isostearyl alcohol, polyoxyethylene sorbitol and sorbitan esters, microcrystalline cellulose, aluminum methoxide and agar, or mixtures of these substances.

[0090] 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 non-aqueous carriers, diluents, solvents or excipients include water, ethanol, polyols and suitable mixtures thereof.

[0091] Dosage forms for topical administration of the compounds of this 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 that may be required.

[0092] The compounds of the present invention may be administered alone or in combination with other pharmaceutically acceptable compounds.

[0093] When using a pharmaceutical composition, a safe and effective amount of the compound of the present invention is administered to a mammal (e.g., a human) in need of treatment, wherein the dosage is a pharmaceutically effective dosage. For a 60 kg human, the daily dosage is generally 1 to 2000 mg, preferably 50 to 1000 mg. Of course, the specific dosage will also take into account factors such as the route of administration and the patient's health condition, all of which are within the skill of a skilled physician.

[0094] The main advantages of the present invention include:

[0095] (a) Unexpectedly, the compound of formula (1) of the present invention exhibits excellent in vitro activity, including NMT1 enzyme inhibitory activity and MV-4-11 cell anti-proliferative activity, and is more suitable for development as a drug.

[0096] (b) The compounds of the present invention have good drugability.

[0097] The features described above, or in the embodiments, may be combined in any combination. All features disclosed in this specification may be used in any combination, and each feature disclosed in this specification may be replaced by any alternative feature that serves the same, equivalent, or similar purpose. Therefore, unless otherwise specified, the features disclosed are merely general examples of equivalent or similar features.

[0098] The following description will elaborate on various specific aspects, characteristics, and advantages of the above-mentioned compounds, methods, and pharmaceutical compositions so that the present invention will be readily apparent. It should be understood that the following detailed description and examples describe specific embodiments and are provided for reference only. After reading the present description, those skilled in the art may make various changes or modifications to the present invention, and such equivalents are within the scope of the present invention.

[0099] The structures of the compounds of the present invention can be confirmed by conventional methods well known to those skilled in the art. If the present invention relates to the absolute configuration of the compounds, the absolute configuration can be confirmed by conventional techniques in the art. For example, single crystal X-ray diffraction (SXRD) is performed by collecting diffraction intensity data on the cultured single crystal using a Bruker D8 venture diffractometer with CuKα phase irradiation as the light source and scanning mode: After scanning and collecting relevant data, the crystal structure is further analyzed using the direct method (Shelxs97) to confirm the absolute configuration.

[0100] The solvents used in the present invention can be obtained commercially. Compounds are named according to conventional nomenclature principles in the art or using ChemDraw@ software. Commercially available compounds use the supplier's catalog names.

[0101] In all embodiments, 1 H-NMR was recorded on a Varian Mercury 400 nuclear magnetic resonance instrument, and chemical shifts are expressed in δ (ppm). Silica gel used for separation was 200-300 mesh unless otherwise specified, and the eluent ratios were all by volume.

[0102] The present invention uses the following abbreviations: (Boc)2O represents di-tert-butyl dicarbonate; CAN(CH3CN) represents acetonitrile; °C represents degrees Celsius; Cs2CO3 represents cesium carbonate; CD3OD represents deuterated methanol; EA(EtOAc) represents ethyl acetate; DCM represents dichloromethane; DIEA(DIPEA) represents diisopropylethylamine; Dioxane represents 1,4-dioxane; DMF represents N,N-dimethylformamide; h represents hour; HATU represents 2-(7-azobenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate; H2O2 represents hydrogen peroxide; K2C O3 stands for potassium carbonate; K3PO4 stands for potassium phosphate; LC-MS stands for liquid chromatography-mass spectrometry; LiAlH4 stands for lithium aluminum tetrahydride; MS stands for mass spectrometry; MsCl stands for methanesulfonyl chloride; NaBH3CN stands for sodium cyanoborohydride; NaHCO3 stands for sodium bicarbonate; NaOH stands for sodium hydroxide; NIS stands for N-iodosuccinimide; NMR stands for nuclear magnetic resonance; Pd2(dba)3 stands for tris(dibenzylideneacetone)dipalladium; Pd(dppf)2Cl2 stands for dichloro[1,1'-bis(diphenylphosphino)ferrocene]palladium; PE stands for petroleum ether; POCl3 stands for phosphorus oxychloride; and TEA stands for triethylamine.

[0103] The present invention will be further described below in conjunction with specific examples. It should be understood that these examples are intended to illustrate the present invention and are not intended to limit the scope of the invention. The experimental methods in the following examples, for which no specific conditions are specified, are generally based on conventional conditions or the conditions recommended by the manufacturer. Unless otherwise stated, percentages and parts are by weight.

[0104] Preparation Example 1: Synthesis of 1,5-dimethyl-4-(2-(methylsulfonyl)oxy)ethyl)-1H-pyrazole-3-carboxylic acid ethyl ester (Intermediate 1)

[0105] Step 1: Synthesis of intermediate 1-1

[0106] Ethyl 1,5-dimethyl-1H-pyrazole-3-carboxylate (5 g, 29.73 mmol) was dissolved in acetonitrile (100 mL) and NIS (7.36 g, 32.70 mmol) was added. The reaction was stirred at 50°C and monitored by LC-MS. The reaction was complete. The acetonitrile was removed by direct concentration and the mixture was poured into water (100 mL). Extraction was performed with EA (50 mL x 3). The organic phases were combined, dried, concentrated, and purified by column chromatography (PE / EA = 20 / 1 to 5 / 1) to afford intermediate 1-1 (8.21 g, 94% yield) as a yellow solid. ESI-MS m / z: 294.9 [M+H] + .

[0107] Step 2: Synthesis of Intermediate 1-2

[0108] Intermediate 1-1 (8.21 g, 27.93 mmol) and potassium vinyl trifluoroborate (4.11 g, 30.72 mmol) were dissolved in dioxane / water (100 / 10 mL). Pd(dppf)Cl2 (2.04 g, 2.79 mmol) and K3PO4 (11.86 g, 55.86 mmol) were added and stirred at 90°C. LC-MS monitoring indicated that the reaction was complete. The dioxane was removed by direct concentration, and the mixture was poured into water (100 mL). Extraction was performed with EA (50 mL x 3). The organic phases were combined, dried, concentrated, and purified by column chromatography (PE / EA = 10 / 1 to 5 / 1) to afford Intermediate 1-2 (4.86 g, 90% yield) as a white solid. ESI-MS m / z: 195.0 [M+H] + .

[0109] Step 3: Synthesis of Intermediates 1-3

[0110] Intermediate 1-2 (4.86 g, 25.05 mmol) was dissolved in THF (50 mL). BH3.THF (1 M, 25.05 mL) was slowly added at 0°C. The reaction was stirred at 0°C for 3 h. 30% aqueous sodium bicarbonate solution (20 mL) and hydrogen peroxide (30%, 20 mL) were then added. The reaction was stirred at 0°C. LC-MS monitoring indicated that the reaction was complete. The mixture was poured into water (100 mL) and extracted with EA (50 mL x 3). The organic phases were combined, dried, concentrated, and purified by column chromatography (PE / EA = 5 / 1 to 1 / 1) to afford Intermediate 1-3 (1.26 g, 24% yield) as a white solid. ESI-MS m / z: 213.0 [M+H] + .

[0111] Step 4: Synthesis of Intermediate 1

[0112] 1-3 (1.26 g, 5.94 mmol) and TEA (1.20 g, 11.89 mmol) were dissolved in DCM (30 mL) under argon protection. MsCl (748 mg, 6.53 mmol) was added with stirring at 0°C. After addition, the reaction was stirred at room temperature. LC-MS monitoring indicated that the reaction was complete. The mixture was poured into water (50 mL) and extracted with DCM (30 mL x 3). The organic phases were combined, dried, concentrated, and purified by column chromatography (PE / EA = 10 / 1 to 2 / 1) to afford intermediate 1 (1.59 g, 92% yield) as a yellow solid. ESI-MS m / z: 291.0 [M+H] + .

[0113] Intermediate 2-4 can be obtained by a similar synthetic method as in Preparation Example 1.

[0114] Preparation Example 2: Synthesis of ethyl 4-(chloromethyl)-1,5-dimethyl-1H-pyrazole-3-carboxylate (Intermediate 5)

[0115] Step 1: Synthesis of Intermediate 5

[0116] Ethyl 1,5-dimethyl-1H-pyrazole-3-carboxylate (2 g, 11.90 mmol) and paraformaldehyde (714 mg, 23.81 mmol) were dissolved in 1,4-dioxane (30 mL). Hydrochloric acid (12 M, 2 mL) and H2SO4 (119 mg, 1.19 mmol, 98%) were added and stirred at 100°C. LC-MS monitoring indicated that the reaction of the starting material was complete. The 1,4-dioxane was removed by direct concentration and the mixture was poured into water (100 mL). The mixture was extracted with EA (50 mL*3). The organic phases were combined, dried, and concentrated to give intermediate 5 (8.21 g, 94% yield) as a yellow solid. ESI-MS m / z: 217.0 [M+H] + .

[0117] Preparation Example 3: Synthesis of ethyl 2-(3-isopropyl-1,5-dimethyl-1H-pyrazol-4-yl)methanesulfonate (Intermediate 6)

[0118] Step 1: Synthesis of Intermediate 6-1

[0119] Methyltriphenylphosphonium bromide (6.21 g, 17.39 mmol) was dissolved in THF (50 mL). Potassium tert-butoxide (1.95 g, 17.39 mmol) was added at room temperature and stirred for 10 minutes. A solution of 1-(1,5-dimethyl-1H-pyrazol-3-yl)ethyl-1-one (2 g, 14.49 mmol) in THF (10 mL) was then added and stirred at room temperature. The reaction was monitored by LC-MS. The reaction was complete. The mixture was poured into water (100 mL) and extracted with EA (50 mL x 3). The organic phases were combined, dried, concentrated, and purified by column chromatography (PE / EA = 20 / 1 to 5 / 1) to afford intermediate 6-1 (1.54 g, 78% yield) as a yellow solid. ESI-MS m / z: 137.0 [M+H] + .

[0120] Step 2: Synthesis of Intermediate 6-2

[0121] Intermediate 6-1 (1.54 g, 11.32 mmol) was dissolved in methanol (50 mL), and Pd / C (150 mg, 10%) was added. The atmosphere was replaced with hydrogen and the reaction was stirred at room temperature. LC-MS monitoring indicated that the reaction was complete. The mixture was directly filtered and concentrated to give Intermediate 6-2 (1.48 g, 95% yield) as a white solid. ESI-MS m / z: 139.0 [M+H] + .

[0122] Subsequently, a similar synthesis method as in Preparation Example 1 was used to obtain intermediate 6.

[0123] Preparation Example 4: Synthesis of ethyl 2-(3-fluoromethyl-1,5-dimethyl-1H-pyrazol-4-yl)methanesulfonate (Intermediate 7)

[0124] Step 1: Synthesis of Intermediate 7-1

[0125] 1,5-Dimethyl-1H-pyrazole-3-methanol (2 g, 15.87 mmol) was dissolved in DCM (60 mL). DAST (3.84 g, 23.81 mmol) was added at room temperature and stirred. The reaction was monitored by LC-MS. The reaction was complete. The mixture was poured into water (100 mL) and extracted with DCM (50 mL x 3). The organic phases were combined, dried, concentrated, and purified by column chromatography (PE / EA = 20 / 1 to 5 / 1) to afford intermediate 7-1 (1.65 g, 82% yield) as a yellow solid. ESI-MS m / z: 129.0 [M+H] + .

[0126] Subsequently, a similar synthesis method as in Preparation Example 1 was used to obtain intermediate 7.

[0127] Preparation Example 5: Synthesis of ethyl 2-(3-cyanomethyl-1,5-dimethyl-1H-pyrazol-4-yl)methanesulfonate (Intermediate 8)

[0128] Step 1: Synthesis of Intermediate 8-1

[0129] 1,5-Dimethyl-1H-pyrazole-3-methanol (2 g, 15.87 mmol) was dissolved in DCM (60 mL). Thionyl chloride (3.78 g, 31.74 mmol) was added at room temperature. The reaction was stirred at room temperature. LC-MS monitoring showed that the reaction of the starting material was complete. Direct concentration gave a yellow solid intermediate 8-1 (2.41 g, yield exceeding 100%). ESI-MS m / z: 145.0 [M+H] + .

[0130] Step 2: Synthesis of Intermediate 8-2

[0131] Intermediate 8-1 (2.41 g, 15.87 mmol) was dissolved in ACN (60 mL). TMSCN (3.15 g, 31.74 mmol) and TBAF (8.30 g, 31.74 mmol) were added at room temperature. The reaction was stirred at 60°C and monitored by LC-MS. The reaction was complete. The mixture was then poured into water (100 mL) and extracted with EA (50 mL x 3). The organic phases were combined, dried, concentrated, and purified by column chromatography (PE / EA = 10 / 1 to 5 / 1) to afford Intermediate 8-2 (1.36 g, 63% yield) as a yellow solid. ESI-MS m / z: 136.0 [M+H] + .

[0132] Subsequently, a similar synthesis method as in Preparation Example 1 was used to obtain intermediate 8.

[0133] Example 1: Synthesis of 4-(2-(5-chloro-2-(3-((dimethylamino)methyl)pyrazolo[1,5-a]pyridin-5-yl)phenoxy)ethyl)-N,N,1,5-tetramethyl-1H-pyrazole-3-carboxamide (Compound 1)

[0134] Step 1: Synthesis of 1-1

[0135] 5-Bromopyrazolo[1,5-A]pyridine (1 g, 5.08 mmol) was dissolved in DMF (20 mL). POCl3 (2.33 g, 15.2 mmol) was slowly added at 0°C. The reaction was stirred at room temperature after addition. LC-MS monitoring indicated that the reaction was complete. The mixture was poured into water (30 mL) and the pH was adjusted to 10 with 1M NaOH. The mixture was extracted with EA (30 mL x 3). The organic phases were combined, dried, and concentrated to afford 1-1 (965 mg, 77% yield) as a white solid. ESI-MS m / z: 224.9 [M+H] + .

[0136] Step 2: Synthesis of 1-2

[0137] 1-1 (200 mg, 0.89 mmol) and 4-chloro-2-hydroxyphenylboronic acid (230 mg, 1.33 mmol) were dissolved in dioxane / water (10 / 2 mL). Pd2(dba)3 (81.4 mg, 0.089 mmol), (Boc)2O (582 mg, 2.67 mmol), and K2CO3 (368 mg, 2.67 mmol) were added. The reaction was stirred at 100°C. LC-MS monitoring indicated that the reaction was complete. Water (30 mL) was added and the mixture was extracted with EA (30 mL*3). The organic phases were combined, dried, concentrated, and purified by column chromatography (PE / EA = 10 / 1 to 3 / 1) to afford 1-2 (211 mg, 87% yield) as a yellow solid. ESI-MS m / z: 272.9 [M+H] + .

[0138] Step 3: Synthesis of 1-3

[0139] 1-2 (100 mg, 0.37 mmol) was dissolved in acetonitrile (10 mL), and intermediate 1 (178 mg, 0.61 mmol) and Cs2CO3 (597 mg, 1.83 mmol) were added. The reaction was stirred at 80°C and monitored by LC-MS. The reaction was complete. Water (30 mL) was added, and the mixture was extracted with EA (30 mL x 3). The organic phases were combined, dried, concentrated, and purified by column chromatography (PE / EA = 20 / 1 to 2 / 1) to afford 2-3 (110 mg, 77% yield) as a yellow solid. ESI-MS m / z: 467.1 [M+H] + .

[0140] Step 4: Synthesis of 1-4

[0141] 1-3 (100 mg, 214 μmol) was dissolved in THF / water (5 / 0.5 mL), and NaOH (12.9 mg, 321 μmol) was added. The reaction was stirred at room temperature and monitored by LC-MS. The reaction was complete. The mixture was poured into water (30 mL), and the pH was adjusted to 5-6 with 1 M hydrochloric acid. The mixture was extracted with EA (30 mL*3). The organic phases were combined, dried, and concentrated to give 1-4 (40 mg, crude) as a yellow solid. ESI-MS m / z: 439.0 [M+H] + .

[0142] Step 5: Synthesis of 1-5

[0143] 1-4 (40 mg, 91.1 μmol) was dissolved in DMF (3 mL), and dimethylamine hydrochloride (14.9 mg, 182 μmol), DIEA (58.9 mg, 456 μmol), and HATU (41.6 mg, 109 μmol) were added sequentially. The reaction was stirred at room temperature and monitored by LC-MS. The reaction was complete. The mixture was poured into water (20 mL) and extracted with EA (10 mL*3). The organic phases were combined, dried, and concentrated to give 1-5 (47 mg, crude) as a yellow solid. ESI-MS m / z: 465.1 [M+H] + .

[0144] Step 6: 1 Synthesis

[0145] 1-5 (47.0 mg, 101 μmol) was dissolved in DCM (5 mL), and TEA (30.6 mg, 303 μmol), NaBH3CN (7.61 mg, 121 μmol), and dimethylamine hydrochloride (24.7 mg, 303 μmol) were added. The reaction was stirred at room temperature. LC-MS monitoring indicated that the reaction was complete. The product was poured into water (20 mL) and extracted with EA (20 mL x 3). The organic phases were combined, dried, and concentrated to afford 1 (14 mg, 28% yield) as a yellow solid.

[0146] 1 H NMR (400MHz, CD3OD): δ8.61(d,J=7.25Hz,1H),8.19(s,1H),7.92-7.95(m,1H),7.44(d,J=8.13Hz,1H),7.17-7.21(m,1H),7.10(br d,J=1.88Hz,2H),4.55-4.61(m,2H),4.20(t,J=6.69Hz,2H),3.68-3.71(m, 3H),2.97-3.07(m,6H),2.93-2.97(m,2H),2.92(s,6H),2.05(s,3H); ESI-MS m / z:495.1[M+H] + .

[0147] Example 2-27: Synthesis of Compounds 2-11, 13-28

[0148] The target compounds 2-11, 13-28 can be obtained by a similar synthesis method as in Example 1.

[0149] Example 28: Synthesis of (4-(2-(5-chloro-2-(3-((dimethylamino)methyl)pyrazolo[1,5-a]pyridin-5-yl)phenoxy)ethyl)-1,5-dimethyl-1H-pyrazol-3-yl)methanol (Compound 12)

[0150] Step 1: Synthesis of 12-1

[0151] 1-3 (200 mg, 0.43 mmol) was dissolved in DCM (10 mL), and TEA (130 mg, 1.29 mmol), NaBH3CN (81 mg, 1.29 mmol), and dimethylamine hydrochloride (105 mg, 1.29 mmol) were added. The reaction was stirred at room temperature. LC-MS monitoring indicated that the reaction was complete. The mixture was poured into water (20 mL) and extracted with EA (20 mL*3). The organic phases were combined, dried, and concentrated to afford 12-1 (220 mg, crude) as a yellow solid. ESI-MS m / z: 496.1 [M+H] + .

[0152] Step 2: 12 Composition

[0153] 12-1 (220 mg, 0.43 mmol) was dissolved in THF (10 mL), and LiAlH4 (33 mg, 0.86 mmol) was added. The reaction was stirred at room temperature. LC-MS monitoring indicated that the reaction was complete. The reaction was quenched with saturated ammonium chloride and extracted with EA (20 mL x 3). The organic phases were combined, dried, and concentrated to afford 12 (84 mg, 43% yield) as a yellow solid.

[0154] 1 H NMR (400MHz, CD3OD): δ8.64(d,J=7.22Hz,1H),8.16(s,1H),7.92-7.97(m,1H),7.41(d,J=8.11Hz,1H),7.22(m,1H),7.12(br d,J=1.82Hz,2H),4.56(m,2H),4.35(s,2H),4.22(t,J=6.61Hz,2H),3.68-3.73(m,3H),3.05(s,6H),2.90(m,2H),2.01(s,3H); ESI-MS m / z:454.1[M+H] + .

[0155] Example 29: Synthesis of 2-(4-(2-(2,3-difluoro-6-(3-((methylamino)methyl)pyrazolo[1,5-a]pyridin-5-yl)phenoxy)ethyl)-1,5-dimethyl-1H-pyrazol-3-yl)propan-2-ol (Compound 29)

[0156] Step 1: Synthesis of 29-1

[0157] 1-1 (500 mg, 2.22 mmol) and 3,4-difluoro-2-methoxyphenylboronic acid (418 mg, 2.22 mmol) were dissolved in dioxane / water (30 mL / 3 mL). Pd2(dba)3 (204 mg, 0.22 mmol) and K2CO3 (613 mg, 4.44 mmol) were added and stirred at 100°C. LC-MS monitoring indicated that the reaction was complete. Water (30 mL) was added and the mixture was extracted with EA (30 mL*3). The organic phases were combined, dried, concentrated, and purified by column chromatography (PE / EA = 10 / 1 to 2 / 1) to afford 29-1 (420 mg, 66% yield) as a yellow solid. ESI-MS m / z: 288.9 [M+H] + .

[0158] Step 2: Synthesis of 29-2

[0159] 29-1 (420 mg, 1.46 mmol) was dissolved in DCM (30 mL), and BBr3 (366 mg, 4.37 mmol) was added at room temperature. The reaction was stirred at room temperature and monitored by LC-MS. The reaction was complete. Water (30 mL) was slowly added to quench the mixture. The mixture was neutralized to a weak base with saturated NaHCO3 and extracted with EA (30 mL x 3). The organic phases were combined, dried, concentrated, and purified by column chromatography (PE / EA = 5 / 1 to 1 / 1) to afford 29-2 (290 mg, 73% yield) as a yellow solid. ESI-MS m / z: 274.9 [M+H] + .

[0160] Step 3: Synthesis of 29-3

[0161] 29-2 (290 mg, 1.06 mmol) was dissolved in DCM (20 mL), and TEA (321 mg, 3.18 mmol), AcOH (64 mg, 1.06 mmol), methylamine hydrochloride (143 mg, 2.12 mmol), and NaBH3CN (333 mg, 5.30 mmol) were added. The reaction was stirred at room temperature. LC-MS monitoring indicated that the starting material had reacted completely. Boc2O (462 mg, 2.12 mmol) was then added, and the reaction was stirred at room temperature. LC-MS monitoring indicated that the starting material had reacted completely. The mixture was poured into water (30 mL) and extracted with DCM (20 mL x 3). The organic phases were combined, dried, concentrated, and purified by column chromatography (PE / EA = 5 / 1 to 1 / 1) to afford 29-3 (310 mg, 75% yield) as a yellow solid. ESI-MS m / z: 390.1 [M+H] + .

[0162] Step 4: Synthesis of 29-4

[0163] 29-3 (310 mg, 0.80 mmol) was dissolved in DMF (30 mL), and intermediate 1 (279 mg, 0.96 mmol) and Cs2CO3 (521 mg, 1.60 mmol) were added. The reaction was stirred at 100°C and monitored by LC-MS. The reaction was complete. Water (30 mL) was added, and the mixture was extracted with EA (30 mL x 3). The organic phases were combined, dried, concentrated, and purified by column chromatography (PE / EA = 20 / 1 to 3 / 1) to afford 29-4 (260 mg, 56% yield) as a yellow solid. ESI-MS m / z: 584.2 [M+H] + .

[0164] Step 5: Synthesis of 29-5

[0165] 29-4 (260 mg, 0.45 mmol) was dissolved in THF (20 mL). Methylmagnesium bromide (1 M, 1.35 mL, 1.35 mmol) was added at -78°C and the reaction mixture was slowly warmed to room temperature with stirring. LC-MS monitoring indicated that the reaction was complete. The mixture was poured into water (30 mL) and extracted with EA (20 mL x 3). The organic phases were combined, dried, concentrated, and purified by column chromatography (PE / EA = 5 / 1 to 1 / 1) to afford 29-5 (130 mg, 51%) as a yellow solid. ESI-MS m / z: 570.2 [M+H] + .

[0166] Step 6: 29 synthesis

[0167] 29-5 (130 mg, 0.23 mmol) was dissolved in DCM (10 mL), and HCl / Dioxane (4 M, 1 mL) was added. The reaction was stirred at room temperature and monitored by LC-MS. The reaction was complete and the mixture was concentrated to give 29 (60 mg, 56% yield) as a yellow solid.

[0168] 1 H NMR (400MHz, DMSO-d6): δ8.66(d,J=7.8Hz,1H),8.18(s,1H),8.03(s,1H),7.41-7.29(m,2H),7.09(dd,J=7.2,1.8Hz,1H),4. 69(s,1H),4.34(s,2H),4.00(t,J=7.8Hz,2H),3.51(s,3H),2.88-2.82(m,2H),2.55(s,3H),1.86(s,3H),1.29(s,6H); ESI-MS m / z:470.1[M+H] + .

[0169] Example 30: Synthesis of Compound 33

[0170] The target compound 33 can be obtained by using a synthetic method similar to that in Example 29.

[0171] Example 31: Synthesis of 1-(4-(2-(2,3-difluoro-6-(3-((methylamino)methyl)pyrazolo[1,5-a]pyridin-5-yl)phenoxy)ethyl)-1,5-dimethyl-1H-pyrazol-3-yl)-2,2-dimethylpropan-1-ol (Compound 30)

[0172] Step 1: Synthesis of 30-1

[0173] 29-4 (200 mg, 0.34 mmol) was dissolved in THF / water (10 / 1 mL), and NaOH (27 mg, 0.69 mmol) was added. The reaction was stirred at room temperature and monitored by LC-MS. The reaction was complete. The mixture was poured into water (30 mL), and the pH was adjusted to 5-6 with 1 M hydrochloric acid. The mixture was extracted with EA (30 mL*3). The organic phases were combined, dried, and concentrated to give 30-1 (170 mg, 90%) as a yellow solid. ESI-MS m / z: 556.1 [M+H] + .

[0174] Step 2: Synthesis of 30-2

[0175] 30-1 (170 mg, 0.31 mmol) was dissolved in DMF (10 mL), and N,O-dimethylhydroxylamine hydrochloride (45 mg, 0.47 mmol), DIEA (80 mg, 0.62 mmol), and HATU (179 mg, 0.47 mmol) were added sequentially. The reaction was stirred at room temperature. LC-MS monitoring indicated that the reaction was complete. The mixture was poured into water (30 mL) and extracted with EA (20 mL x 3). The organic phases were combined, dried, concentrated, and purified by column chromatography (PE / EA = 5 / 1 to 1 / 1) to afford 30-2 (160 mg, 86%) as a yellow solid. ESI-MS m / z: 599.2 [M+H] + .

[0176] Step 3: Synthesis of 30-3

[0177] 30-2 (160 mg, 0.27 mmol) was dissolved in THF (20 mL) and added at -50 °C. t-The mixture was then slowly heated to room temperature with stirring. LC-MS monitoring confirmed the complete reaction. The mixture was poured into water (30 mL) and extracted with EA (20 mL x 3). The organic phases were combined, dried, concentrated, and purified by column chromatography (PE / EA = 5 / 1 to 0 / 1) to afford 30-3 (70 mg, 44%) as a yellow solid. ESI-MS m / z: 598.2 [M+H] + .

[0178] Step 4:30 Synthesis

[0179] 30-3 (70 mg, 0.12 mmol) was dissolved in DCM (10 mL), and HCl / Dioxane (4 M, 1 mL) was added. The reaction was stirred at room temperature. LC-MS monitoring indicated that the reaction was complete. The reaction mixture was directly concentrated to afford 30 (32 mg, 53% yield) as a yellow solid.

[0180] 1 H NMR (400MHz, DMSO-d6): δ8.65(d,J=7.3Hz,1H),8.30(s,1H),8.07(s,1H),7.90(s,1H),7.37-7.30(m,2H),7.03(dd,J=7.2,1.8Hz,1H),4. 10(d,J=7.9Hz,3H),3.86(dd,J=15.7,9.1Hz,2H),2.72-2.57(m,2H),1.87(s,3H),1.23(d,J=6.7Hz,2H),0.89(s,3H),0.78(s,9H); ESI-MS m / z:498.1[M+H] + .

[0181] Example 32: Synthesis of Compound 34

[0182] The target compound 34 can be obtained by using a similar synthetic method as in Example 30.

[0183] Example 33: Synthesis of 1-(4-(2-(2,3-difluoro-6-(3-((methylamino)methyl)pyrazolo[1,5-a]pyridin-5-yl)phenoxy)ethyl)-1,5-dimethyl-1H-pyrazol-3-yl)ethan-1-ol (Compound 31)

[0184] Step 1: Synthesis of 31-1

[0185] 30-2 (200 mg, 0.33 mmol) was dissolved in THF (20 mL). Methylmagnesium bromide (1 M, 0.40 mL, 0.40 mmol) was added at -78°C and the mixture was slowly warmed to room temperature with stirring. LC-MS monitoring indicated that the reaction was complete. The mixture was poured into water (30 mL) and extracted with EA (20 mL x 3). The organic phases were combined, dried, concentrated, and purified by column chromatography (PE / EA = 5 / 1 to 1 / 1) to afford 31-1 (110 mg, 59%) as a yellow solid. ESI-MS m / z: 554.2 [M+H] + .

[0186] Step 2: Synthesis of 31-2

[0187] 31-1 (110 mg, 0.20 mmol) was dissolved in methanol (10 mL). NaBH4 (11 mg, 0.30 mmol) was added at room temperature and stirred. The reaction was monitored by LC-MS. The reaction was complete. The mixture was poured into water (30 mL) and extracted with EA (20 mL x 3). The organic phases were combined, dried, concentrated, and purified by column chromatography (PE / EA = 5 / 1 to 0 / 1) to give 31-2 (85 mg, 77%) as a yellow solid. ESI-MS m / z: 556.2 [M+H] + .

[0188] Step 3: Synthesis of 31

[0189] 31-2 (85 mg, 0.15 mmol) was dissolved in DCM (10 mL), and HCl / Dioxane (4 M, 1 mL) was added. The reaction was stirred at room temperature and monitored by LC-MS. The reaction was complete and the mixture was concentrated to afford 31 (22 mg, 33% yield) as a yellow solid.

[0190] 1 H NMR (400MHz, DMSO-d6): δ8.61(d,J=7.7Hz,1H),8.19(s,1H),8.05(s,1H),7.41-7.26(m,2H),7.11(m,1H),4.66(s,1H),4.3 1(s,2H),4.01(t,J=7.8Hz,2H),3.81(m,1H),3.52(s,3H),2.88-2.80(m,2H),2.57(s,3H),1.88(s,3H),1.25(s,3H); ESI-MS m / z:446.1[M+H] + .

[0191] Examples 34-36: Synthesis of Compounds 32, 35, and 36

[0192] Using a similar synthetic method as in Example 31, target compounds 32, 35, and 36 can be obtained.

[0193] Example 37: Screening of compounds for NMT1 enzyme catalytic inhibitory activity

[0194] This study evaluated the ability of the compounds of the present invention to inhibit the catalytic activity of NMT1 in vitro. This experiment used FI fluorescence intensity analysis to characterize NMT1 inhibitors in the presence of 1.25 μM myristoyCoA (myristyl CoA lithium salt, Sigma, M4414). In this experiment, a peptide HSPP60SRC (Nanjing Peptide Industry, NJP23953) with the sequence H-Gly-Ser-Asn-Lys-Ser-Lys-Pro-Lys-NH2 was synthesized as the reaction substrate. The final concentrations of NMT1 enzyme were 6 nM, CPM (reactive thiol fluorescent probe, Invitrogen, D346) were 1 μM, HSPP60 SRC were 20 μM, and DMSO were 0.5%.

[0195] Compounds were dissolved in DMSO to obtain a 10 mM stock solution. Dose gradient reactions were prepared with an endpoint compound concentration of 3 mM and then three-fold dilutions were performed in DMSO for a total of ten data points. Using an ECHO acoustic pipetting device (BECKMAN, ECHO 655 system), 0.05 μL of the diluted compound solution was transferred to a 384-well assay plate (Corning, CLS4514). 2.5 μL of NMT1 enzyme working solution was added to the 384-well assay plate and incubated at 25°C for 10 minutes. Then, 5 μL of CPM and myristoy COA working solution was added, and the reaction was centrifuged at 1000 rpm for 1 minute. Subsequently, 2.5 μL of HSPP60 SRC working solution was added and the reaction was incubated at 25°C for 45 minutes. FI signals (em: 320 nm; ex: 405 nm) were detected using an HTS high-throughput drug screening multi-function microplate reader (BMG, PHERA star FSX).

[0196] The inhibition rate of the compound on the catalytic activity of NMT1 enzyme was calculated using the following formula: Inhibition percentage (%) = 100*(mean value of DMSO group - mean value of compound) / (mean value of DMSO group - mean value of blank control group). The IC value of the compound was fitted according to the nonlinear regression equation using the software XLfit 5.5.0. 50 Table 1 provides the inhibitory activity of the compounds of the present invention on NMT1 enzyme catalysis.

[0197] Example 38: Screening of antiproliferative activity of compounds against MV-4-11 cells

[0198] This study used a cell proliferation assay to analyze the cytotoxicity of MV-4-11 human myelomonocytic leukemia cells after three days of treatment with an NMT1 inhibitor. The MV-4-11 cell line was purchased from Nanjing Kebai Biotechnology Co., Ltd. and cultured in IMDM medium (Viva Cell) at 37°C in a cell culture incubator (Thermo) with 5% CO2. For this experiment, the NMT1 inhibitor was dissolved in DMSO to obtain a 600 μM stock solution. Dose gradient reactions were prepared with a 3 μM endpoint compound concentration and three-fold dilutions were performed in DMSO for a total of eight data points. The final DMSO concentration was 0.5%.

[0199] MV-4-11 cells were seeded in a white 96-well plate with 80 μL of cell suspension per well, containing 6,000 MV-4-11 cells. The plate was incubated overnight in a CO2 incubator. Then, 20 μL of NMT1 inhibitor solution at varying concentrations was added to the plate, and the 96-well plate was incubated in the incubator for three days. A separate plate was prepared, and the signal value was read on the day of drug addition, which was used as the maximum value (Max value in the equation below) for data analysis. 25 μL of cell viability chemiluminescent detection reagent was added to each well of this plate, and the plate was incubated at room temperature for 10 minutes to stabilize the luminescent signal. The readings were read using a multilabel analyzer.

[0200] The raw data were converted into inhibition rate, IC, using the equation (Sample-Min) / (Max-Min)*100%. 50 The value can be obtained by four-parameter curve fitting (derived using the "log (inhibitor) vs. response -- Variable slope" mode in GraphPad Prism). Min: 0.5% DMSO-treated wells; Max: Day 0 wells. Table 1 provides the inhibitory activity of the compounds of the present invention on MV-4-11 cell proliferation.

[0201] Table 1: Inhibitory activity of compounds against NMT1 enzyme (IC 50 ) and antiproliferative activity against MV-4-11 cells (IC 50 )

[0202] From the data in the above table, it can be seen that the compounds of the present invention have strong inhibitory activity against NMT1 enzyme catalysis and strong anti-proliferative activity against MV-4-11 cells. In particular, compounds 21, 27, 29, 30, 31, 32, 34 and 36 have strong inhibitory activity against NMT1 enzyme catalysis and anti-proliferative activity against MV-4-11 cells. The IC inhibitory activity against NMT1 enzyme catalysis is 50 Less than 2nM, IC value for antiproliferative activity against MV-4-11 cells 50 Less than 10nM.

[0203] All documents mentioned in this application are incorporated herein by reference, just as if each document were incorporated herein by reference individually. It should also be understood that after reading the above teachings of the present invention, those skilled in the art may make various changes or modifications to the present invention, and that such equivalents also fall within the scope of the claims appended hereto.

Claims

1. A compound having a structure as shown in formula (1), or its isomer, crystal form, pharmaceutically acceptable salt, hydrate or solvate: In formula (1): n is 0, 1, 2, or 3; X 1 and X 2 Each independently selected from the following group: CH, CR 1 or N; W is selected from the group consisting of a chemical bond, O, S or NH; Y is selected from the group consisting of: chemical bonds, -(CH2) m -、-(CH2) m O- or -(CH2) m O-(CH2) m , wherein m is selected from 1, 2 or 3; L is -(CH2) m -; Each R 1 Each independently selected from the group consisting of D, hydroxyl, halogen, CN, NO2, NH2, C 1-4 Alkyl, C 2-4 Alkenyl, C 2-4 Alkynyl, C 1-4 Haloalkyl, C 1-4 Alkoxy, C 1-4 Halogenated alkoxy, C 3-6 alkyl, saturated or partially unsaturated carbocyclic group, 3-8 membered saturated or partially unsaturated heterocyclic group, C 6-10 Aryl or 5-12 membered heteroaryl; R 2 For-NR 6 R 7 , where R 6 and R 7 Each independently selected from the following group: H, D, C 1-6 Alkyl, C 1-6 Haloalkyl; Or, R 6 and R 7 and the nitrogen atom to which it is connected together form a 3-8 membered saturated or partially unsaturated heterocyclic group, or a 5-12 membered heteroaryl group; R 3 Selected from the following group: H, D, C 1-4 Alkyl, C 1-4 Haloalkyl, C 1-4 Alkoxy, C 1-4 Halogenated alkoxy, C 3-6 alkyl, saturated or partially unsaturated carbocyclic group, 3-8 membered saturated or partially unsaturated heterocyclic group, C 6-10 Aryl or 5-12 membered heteroaryl; R 4 Select from the following group: C 1-4 Alkyl, C 1-4 Haloalkyl, C 3-6 alkyl, saturated or partially unsaturated carbocyclic group, 3-8 membered saturated or partially unsaturated heterocyclic group, C 6-10 Aryl or 5-12 membered heteroaryl; R 5 Selected from the group consisting of hydroxyl, CN, C 1-6 Alkyl, C 1-6 Halogenated alkyl, hydroxy substituted C 1-6 Alkyl, cyano substituted C 1-6 Alkyl or -C(O)NR 8 R 9 ; R 8 and R 9 Each independently selected from the following group: H, D, C 1-6 Alkyl, C 1-6 Haloalkyl; Or, R 8 and R 9 and the nitrogen atom to which it is connected together form a 3-8 membered saturated or partially unsaturated heterocyclic group, or a 5-12 membered heteroaryl group; in, The alkyl, alkenyl, alkynyl, halogenated alkyl, alkoxy, halogenated alkoxy, carbocyclic The radical, heterocyclic radical, aryl radical and heteroaryl radical are each independently optionally substituted by one or more substituents selected from the group consisting of D, hydroxyl, halogen, CN, NO2, NH2, C 1-4 Alkyl, C 2-4 Alkenyl, C 2-4 Alkynyl, C 1-4 Haloalkyl, C 1-4 Alkoxy or C 1-4 Halogenated alkoxy.

2. The compound according to claim 1, or its isomer, crystal form, pharmaceutically acceptable salt, hydrate or solvate, wherein in the formula (1), each R 1 Each independently selected from the following group: F, Cl, Br, CN, NO2, C 1-3 Alkyl, C 1-3 Haloalkyl, C 1-3 Alkoxy or C 1-3 Halogenated alkoxy.

3. The compound according to claim 1, or its isomer, crystal form, pharmaceutically acceptable salt, hydrate or solvate, wherein in the formula (1), R 2 For-NR 6 R 7 , where R 6 and R 7 Each independently selected from the following group: H, D, C 1-3 Alkyl, C 1-3 Halogenated alkyl.

4. The compound according to claim 1, or its isomer, crystal form, pharmaceutically acceptable salt, hydrate or solvate, wherein in the formula (1), R 3 Selected from the following group: H, D, C 1-3 Alkyl, C 1-3 Halogenated alkyl.

5. The compound according to claim 1, or its isomer, crystal form, pharmaceutically acceptable salt, hydrate or solvate, wherein in the formula (1), R 4 Select from the following group: C 1-3 Alkyl, C 1-3 Halogenated alkyl.

6. The compound according to claim 1, or its isomer, crystal form, pharmaceutically acceptable salt, hydrate or solvate, wherein in the formula (1), R 5 Selected from the following group: CN, C 1-6 Alkyl, C 1-6 Halogenated alkyl, hydroxy substituted C 1-6 Alkyl, cyano substituted C 1-6 Alkyl or -C(O)NR 8 R 9 ; in, R 8 and R 9 Each independently selected from the following group: H, D, C 1-3 Alkyl, C 1-3 Halogenated alkyl.

7. The compound, its isomer, crystalline form or pharmaceutically acceptable salt thereof according to claim 1, wherein the compound has a structure selected from the group consisting of:

8. A pharmaceutical composition for treating, regulating and / or preventing diseases mediated by NMT, characterized in that: The pharmaceutical composition comprises: (1) a compound according to any one of claims 1 to 7, or its isomers, crystal forms, pharmaceutically acceptable salts, hydrates or solvates as an active ingredient; and Optional (2) a pharmaceutically acceptable excipient or carrier.

9. Use of the compound according to any one of claims 1 to 7, its isomers, crystal forms, pharmaceutically acceptable salts, hydrates or solvates, characterized in that: Used for preparing drugs for treating, regulating and / or preventing diseases mediated by NMT.

10. A method for treating, regulating and / or preventing diseases mediated by NMT, characterized in that: The method comprises the steps of administering to an individual in need thereof the compound according to any one of claims 1 to 7, or its isomers, crystal forms, pharmaceutically acceptable salts, hydrates or solvates, or the pharmaceutical composition according to claim 8.

11. The use according to claim 9 or the method according to claim 10, characterized in that The diseases mediated by NMT include infectious diseases or hyperproliferative diseases.

12. The use or method according to claim 11, characterized in that The infectious diseases include protozoan infections and viral infections; preferably, the protozoan infections include malaria and leishmaniasis; preferably, the viral infections include human rhinovirus infections and HIV infections.

13. The use or method according to claim 11, characterized in that The hyperproliferative disease is selected from the group consisting of lymphoma, leukemia, brain tumor, gastric cancer, liver cancer, lung cancer, intestinal cancer, pancreatic cancer, breast cancer, cervical cancer, ovarian cancer, endometrial cancer and prostate cancer.

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