A deuterated pyridinone compound, and a preparation method and application thereof

Abstract

The present application belongs to the technical field of chemical drugs, and relates to a compound shown in a general formula (I), or a racemate thereof, or an isomer thereof and a pharmaceutically acceptable salt thereof, as a PCSK9 inhibitor and a method for treating various specific diseases or conditions using the same.

Description

TECHNICAL FIELD

[0001] The present application belongs to the technical field of chemical drugs, and relates to a deuterated pyridone compound, or an isomer thereof, or a racemate thereof, or a pharmaceutically acceptable salt thereof, and a preparation method and application thereof. As a PCSK9 inhibitor, and a method for treating various specific diseases or conditions using the same. BACKGROUND

[0002] Proprotein convertase subtilisin / kexin type 9 (PCSK9), also known as NARC-1, is a proprotein-converting enzyme in the subtilisin (S8) family of serine proteases, which is expressed in cells with proliferative and differentiative capacity, including hepatocytes, renal interstitial cells, ileal and colonic epithelial cells, and embryonic hindbrain neurons. Studies have found that PCSK9 plays a role in the differentiation of hepatocytes and neural cells, which can not only specifically act on cholesterol biosynthesis or uptake, but also circulating PCSK9 can directly bind to the low-density lipoprotein receptor (LDLR) on the surface of hepatocytes, be phagocytosed with LDLR by hepatocytes, promote the degradation of LDLR in hepatocytes, hinder the circulation of LDLR, thereby increasing the content of LDL cholesterol (LDL-C) in the blood plasma, and the expression of LDL-C is closely related to human dyslipidemia and cardiovascular diseases.

[0003] Currently, researches on inhibiting the function of PCSK9 or inhibiting the generation of PCSK9 are being carried out. For example, attempts such as inhibiting the function of PCSK9 with monoclonal antibodies targeting PCSK9, inhibiting the generation of PCSK9 by RNA interference, etc. have been reported. However, for patients with cardiovascular diseases, effective small molecule inhibitors are needed to inhibit the function of PCSK9. SUMMARY

[0004] In view of the problems existing in the prior art, the present application provides a compound represented by general formula (I), or an isomer thereof, or a racemate thereof, or a pharmaceutically acceptable salt thereof, and a preparation method and application thereof. As a PCSK9 inhibitor, and a method for treating various specific diseases or conditions using the same.

[0005] In a first aspect, the present application provides a compound represented by general formula (I), or an isomer thereof, or a racemate thereof, or a pharmaceutically acceptable salt thereof:

[0006]

[0007] In a second aspect, the present application further provides a pharmaceutical composition comprising a therapeutically effective amount of the compound or the pharmaceutically acceptable salt thereof according to any one of the above and a pharmaceutically acceptable carrier.

[0008] In a third aspect, the present invention also provides a use of a therapeutically effective amount of the above-mentioned compound or a pharmaceutically acceptable salt thereof in the preparation of a medicament for treating a disease condition, wherein the disease is a PCSK9 inhibitor-related disease, specifically, the disease condition is selected from hypercholesterolemia and the like.

[0009] Specifically, the present invention is achieved through the following technical solutions:

[0010] A compound represented by general formula (I), or its isomer, racemate, or pharmaceutically acceptable salt, comprising:

[0011]

[0012] Among them, R1, R2, R3, R4, R5, R6, R7, R8, R9, R 10 、R 11 、R 12 、R 13 、R 14 、R 15 、R 16 、R 17 、R 18 、R 19 、R 20 Each is selected from hydrogen or deuterium; at the same time, R1, R2, R3, R4, R5, R6, R7, R8, R9, R 10 、R 11 、R 12 、R 13 、R 14 、R 15 、R 16 、R 17 、R 18 、R 19 、R 20 At least one is selected from deuterium.

[0013] As a preferred technical solution of the present invention, R1 is selected from deuterium, R2, R3, R4, R5, R6, R7, R8, R9, R 10 、R 11 、R 12 、R 13 、R 14 、R 15 、R 16 、R 17 、R 18 、R 19 、R 20 are each selected from hydrogen or deuterium.

[0014] As a preferred technical solution of the present application, R5 is selected from deuterium, R1, R2, R3, R4, R6, R7, R8, R9, R 10 11 12 13 14 15 16 17 18 19 20 Each of R1, R2, R3, R4, R5, R6, R7, R8, R9, R

[0015] As a preferred technical solution of the present application, one or all of R 18 19 Each of R1, R2, R3, R4, R5, R6, R7, R8, R9, R 10 11 12 13 14 15 16 17 20 Each of R1, R2, R3, R4, R5, R6, R7, R8, R9, R

[0016] As a preferred technical solution of the present application, the compound, or its isomer, or its racemate, or its pharmaceutically acceptable salt is selected from:

[0017]

[0018]

[0019] As a preferred technical solution of the present application, the pharmaceutically acceptable salt refers to the compound, or its isomer, or its racemate, or its pharmaceutically acceptable salt prepared with a pharmaceutically acceptable acid or base.

[0020] The present application further provides a pharmaceutical composition, characterized in comprising a therapeutically effective amount of the compound, or its isomer, or its racemate, or its pharmaceutically acceptable salt and a pharmaceutically acceptable carrier.

[0021] The present application further provides the medical use of the compound, or its isomer, or its racemate, or its pharmaceutically acceptable salt, in particular, the use in the preparation of a drug for treating diseases, which are related to PCSK9 inhibitors, and are particularly selected from hypercholesterolemia and other diseases.

[0022] For the sake of clarity, general terms used in the description of the compounds are defined herein.

[0023] ​​​​​​​​​​​​​​​​​​​The following terms and phrases, as used herein, are intended to have the following meanings unless otherwise indicated. A particular term or phrase should not be construed to be indefinite or unclear in the absence of a specific definition, but should be interpreted in accordance with its ordinary meaning. Where a trade name appears herein, the corresponding generic or active ingredient is intended to be a substitute therefor. The term "pharmaceutically acceptable" is employed herein to describe 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 problem or complication, commensurate with a reasonable benefit / risk ratio.

[0024] The term "pharmaceutically acceptable salt" means a salt of a compound of the present application, prepared from a compound of the present application having specific substituents discovered herein with a pharmaceutically acceptable acid or base.

[0025] In addition to salt forms, the compounds of the present application can exist in a prodrug form. Prodrugs of the compounds described herein readily undergo chemical changes under physiological conditions to form the compounds of the present application. Additionally, prodrugs can be converted to the compounds of the present application by chemical or biochemical methods in an ex vivo environment.

[0026] Certain compounds of the present application can exist in unsolvated as well as solvated forms, including hydrated forms. In general, the solvated forms are equivalent to unsolvated forms and are intended to be encompassed within the scope of the present application.

[0027] The compounds of the present application can exist in particular geometric or stereoisomeric forms. The present application contemplates all such compounds, including cis- and trans-isomers, (-)- and (+)-enantiomers, (R)- and (S)-enantiomers, diastereomers, (D)- isomers, (L)-isomers, atropisomers, racemic mixtures, and mixtures thereof, as falling within the scope of the present application. Additional asymmetric carbon atoms can be present in a substituent group. All such isomers, as well as mixtures thereof, are included within the scope of the present application.

[0028] Optically active (R)- and (S)-isomers, as well as D and L isomers, atropisomers, and the like can be prepared by chiral synthesis or chiral reagents or other conventional techniques. If one enantiomer of a compound of the application 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 cleaved to yield the pure desired enantiomer. Alternatively, when a basic functional group (such as an amino group) or an acidic functional group (such as a carboxyl group) is present in the molecule, a diastereomeric salt form of the compound with an appropriate optically active acid or base can be formed, and the diastereomeric salt separated by conventional means, and the desired enantiomer recovered by treating with base or acid, as appropriate. Additionally, separation of the enantiomers and diastereomers is typically accomplished by chromatography using a chiral stationary phase, optionally in combination with chemical derivatization (e.g., formation of a carbamate from an amine).

[0029] The atoms of the molecules of the compounds of the application can be isotopically enriched. Isotopic enrichment often can extend half-life, decrease clearance, increase metabolic stability, and increase in vivo activity, among other effects. Also, included is an embodiment wherein at least one atom is replaced by an atom having the same atomic number (number of protons) and a different mass number (number of protons and neutrons). Examples of isotopes of atoms that are included in the compounds of the application include hydrogen, carbon, nitrogen, oxygen, phosphorus, sulfur, fluorine, chlorine, which respectively include 2 H, 3 H, 13 C, 14 C, 15 N, 17 O, 18 O, 31 p, 32 P, 35 S, 18 F, 36 Cl. In particular, radioactive isotopes such as 3 H or 14 C that emit radiation while disintegrating can be used for drug preparation or for localizing compounds in the body in situ. Stable isotopes neither disintegrate nor emit radiation, and thus can be used safely. When the atoms that make up the molecules of the compounds of the application are isotopes, isotopes can be converted according to general methods by replacing reagents used in the synthesis with reagents containing the corresponding isotope.

[0030] The compounds of the application can contain unnatural proportions of atomic isotopes at one or more of the atoms that constitute such compounds. For example, the compounds can be radiolabeled with radioactive isotopes, such as for example deuterium ( 2 H), iodine-125 ( 125 I) or C-14 ( 14C). All isotopic variations of the compounds of the present application, whether radioactive or not, are included within the scope of the present application.

[0031] Further, the present application includes compounds of the present application wherein one or more hydrogen atom has been replaced by a deuterium isotope ( 2 H)substituted, the deuterium substituted compounds of the present application have the effect of increasing half-life, decreasing clearance, metabolic stability and increasing in vivo activity.

[0032] The method of preparing the isotopic derivatives generally involves a phase transfer catalysis method. For example, a preferred method of deuterium incorporation employs a phase transfer catalyst (e.g., a tetraalkylammonium salt, NBu4HSO4). The use of a phase transfer catalyst to exchange the methylene protons of diphenylmethane compounds results in higher deuterium incorporation than is achieved by reduction with a deuterated silane (e.g., triethyldeuteratedsilane) or with sodium borodeuteride in the presence of an acid (e.g., methanesulfonic acid) using a Lewis acid such as aluminum trichloride.

[0033] The term "pharmaceutically acceptable carrier" means any formulation carrier or medium that does not interfere with the biological activity of the active substance and that is nontoxic to the host or patient in which it is administered. Representative carriers include water, oil, vegetable and mineral, cream bases, lotion bases, ointment bases, and the like. These bases include suspending agents, viscosity increasing agents, penetration enhancers, and the like. Their formulation is well known to those skilled in the art of cosmetics or topical pharmaceuticals. Additional information on carriers can be found in Remington: The Science and Practice of Pharmacy, 21st Ed., Lippincott, Williams & Wilkins (2005), the contents of which are incorporated herein by reference.

[0034] The term "excipient" generally refers to a carrier, diluent and / or vehicle with which an active agent is formulated to facilitate administration of the active agent.

[0035] The term "effective amount" or "therapeutically effective amount" with respect to a pharmaceutical or pharmacological agent means a sufficient amount of the agent to achieve the intended effect without being toxic to the recipient. With respect to oral dosage forms of the present application, an "effective amount" of one active agent in a composition means the amount of the agent that is required to achieve the intended effect in combination with another active agent in the composition. The determination of an effective amount will vary from subject to subject, depending on the age and general condition of the recipient, as well as the particular active agent involved. An appropriate effective amount for a given situation will be ascertainable by one of ordinary skill in the art using only routine experimentation.

[0036] The term "active ingredient", "therapeutic agent", "active agent" or "active substance" means a chemical entity that is effective in treating a target disorder, disease or condition.

[0037] "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.

[0038] The compounds of the present invention can be prepared by a variety of synthetic methods well known to those skilled in the art, including the specific embodiments listed below, embodiments formed by combining them with other chemical synthesis methods, and equivalent substitutions well known to those skilled in the art. Preferred embodiments include but are not limited to the examples of the present invention. DETAILED DESCRIPTION

[0039] The present application is further described in detail below with reference to examples, but the implementation methods of the present application are not limited thereto.

[0040] Example 1

[0041] Synthesis of 6-(5-(difluoromethoxy)pyrimidin-2-amino)cyclopentyl)amino)-2H-[1,3′-bipyridyl]-2-one-3-d

[0042]

[0043] Synthesis route:

[0044]

[0045] Step A: Pyridin-2(1H)-one-3-d

[0046]

[0047] At room temperature, 3-bromopyridin-2(1H)one (1.0 g, N / A) was added to deuterated methanol (30 ml), and then Pd / C (200 mg, 55% water content) was added to the system. The system was replaced with nitrogen twice, and then replaced with a deuterium balloon three times, and the reaction was carried out at room temperature for 12 hours.

[0048] After the reaction is complete, the mixture is filtered and the resulting clear solution is directly dried by rotary evaporation. An appropriate amount of methanol is then added for dissolution and the mixture is concentrated to dryness to obtain 500 mg of a pink solid product, pyridin-2(1H)-one-3-d (yield 89%). LC-MS: [M+H] + =97.

[0049] Step B: tert-Butyl (1S,3S)-3-((5-(difluoromethoxy)pyrimidin-2-amino)cyclopentyl)carbamate

[0050]

[0051] To a solution of (1S,3S)-3-((5-(difluoromethoxy)pyrimidin-2- yl)amino)cyclopentyl)carbamic acid tert-butyl ester (1.7 g, 4.53 mmol) in hydrochloric acid / ethyl acetate (2 M, 20 mL) was added at room temperature. The reaction was stirred at room temperature for 5 h.

[0052] After the reaction was completed, the mixture was cooled to room temperature, water (40 mL) was added, and the mixture was extracted with ethyl acetate (20 mL x 3). The organic phase was combined, washed with saturated brine (30 mL), dried, filtered, and concentrated to dryness under reduced pressure to give a residue. The residue was purified by normal phase column (n-hexane / ethyl acetate = 1:1) to give 1.7 g of (1S,3S)-3-((5-(difluoromethoxy)pyrimidin-2-yl)amino)cyclopentyl)carbamic acid tert-butyl ester (yield 89%) as a white solid. LC-MS: [M+H] + = 345.

[0053] Step C: (1S,3S)-N1-(5-(difluoromethoxy)pyrimidin-2-yl)cyclopentane-1,3-diamine hydrochloride

[0054]

[0055] To a solution of (1S,3S)-3-((5-(difluoromethoxy)pyrimidin-2- yl)amino)cyclopentyl)carbamic acid tert-butyl ester (1.7 g, 4.53 mmol) in hydrochloric acid / ethyl acetate (2 M, 20 mL) was added at room temperature. The reaction was stirred at room temperature for 5 h.

[0056] After the reaction was completed, the mixture was cooled to room temperature, water (40 mL) was added, and the mixture was extracted with ethyl acetate (20 mL x 3). The organic phase was combined, washed with saturated brine (30 mL), dried, filtered, and concentrated to dryness under reduced pressure to give a residue. The residue was purified by normal phase column (n-hexane / ethyl acetate = 1:1) to give 1.7 g of (1S,3S)-3-((5-(difluoromethoxy)pyrimidin-2-yl)amino)cyclopentyl)carbamic acid tert-butyl ester (yield 89%) as a white solid. LC-MS: [M+H] + = 245.

[0057] Step D: (1S,3S)-N1-(5-(difluoromethoxy)pyrimidin-2-yl)-N3-(5-iodopyridin-2-yl)cyclopentane-1,3-diamine

[0058]

[0059] To a solution of (1S,3S)-N1-(5-(difluoromethoxy)pyrimidin-2-yl)cyclopentane-1,3-diamine hydrochloride (1.27 g, 4.53 mmol) and 2-fluoro-5-iodopyridine (1.01 g, 4.53 mmol) in DMSO (20 mL) was added potassium carbonate (3.12 g, 22.65 mmol) at room temperature. The reaction was stirred at 140 °C for 12 h.

[0060] The reaction was completed and cooled to room temperature, water (40 mL) was added to the system, extracted with ethyl acetate (20 mL x 3). The organic phase was combined, washed with saturated brine (30 mL), dried, filtered and concentrated to dryness under reduced pressure to give a residue. The residue was purified by normal phase column (n-hexane / ethyl acetate = 1 :2) to give 1.23 g of white product (1S,3S)-N1-(5-difluoromethoxy)pyrimidin-2-yl)-N3-(5-iodopyridin-2-yl)cyclopentane-1,3-diamine (yield 60%). LC-MS: [M+H]+= 448.

[0061] Step E: 6-(5-(difluoromethoxy)pyrimidin-2-ylamino)cyclopentyl)amino)-2H- [1,3'-bipyridinyl]-2-one-3-d

[0062]

[0063] (1S,3S)-N1-(5-difluoromethoxy)pyrimidin-2-yl)-N3-(5-iodopyridin-2-yl)cyclopentane-1,3-diamine (300 mg, 0.67 mmol) and pyridin-2(1H)-one-3-d (130 mg, 1.34 mmol), (1S,2S)-N1,N2-dimethylcyclohexane-1,2-diamine (19.0 mg, 0.13 mmol), potassium phosphate (284.5 mg, 1.34 mmol) were added to DMSO (8 mL), followed by the addition of cuprous iodide (38.2 mg, 0.20 mmol) at room temperature. After the addition, the system was replaced with nitrogen and raised to 140 degrees Celsius for 12 hours.

[0064] The reaction was completed and cooled to room temperature, water (40 mL) was added to the system, extracted with ethyl acetate (20 mL x 3). The organic phase was combined, washed with saturated brine (30 mL), dried, filtered and concentrated to dryness under reduced pressure to give a residue. The residue was purified by normal phase column (n-hexane / ethyl acetate = 1 :2) to give 1.23 g of white product (1S,3S)-N1-(5-difluoromethoxy)pyrimidin-2-yl)-N3-(5-iodopyridin-2-yl)cyclopentane-1,3-diamine (yield 60%). LC-MS: [M+H]+= 448.

[0065] LC-MS: [M+H]+= 416.

[0066] 1H NMR (400 MHz, DMSO-d6) δ 8.25-8.22 (s, 2H), 7.93-7.90 (d, J = 2.6 Hz, 1H), 7.62-7.58 (dd, J = 6.8, 2.1 Hz, 1H), 7.52-7.45 (m, 2H), 7.41-7.37 (dd, J = 8.9, 2.7 Hz, 1H), 7.23-6.84 (m, 2H), 6.55-6.50 (d, J = 8.9 Hz, 1H), 6.29-6.25 (t, J = 6.7 Hz, 1H), 4.36-4.26 (p, J = 6.7 Hz, 2H), 2.18-2.07 (m, 2H), 1.95-1.81 (m, 2H), 1.59-1.45 (m, 2H).

[0067] Example 2

[0068] Synthesis of 6'-(((1S,3S)-3-((5-(difluoromethoxy)pyrimidin-2-yl)amino)cyclopentyl)amino)- 2H-[1,3'-bipyridinyl]-2-one-2'-d

[0069]

[0070] Synthesis route:

[0071]

[0072] Step A: tert-butyl (1s,3S)-3-((6-bromopyridin-2-yl)amino)cyclopentyl)carbamate

[0073]

[0074] At room temperature, 2-bromo-6-fluoropyridine (1 g, 5.68 mmol), tert-butyl (1S,3S)-3- aminocyclopentyl)carbamate (1.14 g, 5.68 mmol) were dissolved in DMSO (10 ml), potassium carbonate (3.92 g, 28.4 mmol) was added, the reaction system was slowly warmed to 140 °C under nitrogen protection, and the reaction was carried out overnight.

[0075] After the reaction was completed, water (10 ml) was added, and the organic phase was extracted with ethyl acetate (60 ml x 3). The organic phase was combined and concentrated under reduced pressure, and the residue was purified by silica gel column (DCM / MeOH = 10:1) to obtain 1.6 g of light yellow oily liquid product (1S,3S)-3-((6-bromopyridin-2-yl)amino)cyclopentyl)carbamate (yield 73%). LC-MS: [M+H]+= 356.

[0076] Step B: Synthesis of tert-butyl (1S,3S)-3-((pyridin-2-yl-6-amino)cyclopentyl)carbamate

[0077]

[0078] (1S,3S)-3-((6-bromopyridin-2-yl)amino)cyclopentyl)carbamic acid tert-butyl ester (1.6 g, 4.49 mmol) was dissolved in deuterated methanol (20 mL) and heavy water (20 mL) at room temperature, Pd / C (320 mg, 10% v / w) was added, and the reaction was stirred at room temperature for 8 hours under deuterium gas protection.

[0079] After the reaction was completed, the mixture was filtered, and the filter cake was washed with methanol (20 mL). The filtrate was concentrated to dryness to give (1S,3S)-3-((pyridin-2-yl-6-amino)cyclopentyl)carbamic acid tert-butyl ester as a light yellow solid (700 mg). The product was used directly in the next step without further purification. LC-MS: [M+H]+ = 279. +

[0080] Step C: Synthesis of (1S,3S)-3-((5-iodopyridin-2-yl-6-amino)cyclopentyl)carbamic acid tert-butyl ester

[0081]

[0082] (1S,3S)-3-((pyridin-2-yl-6-amino)cyclopentyl)carbamic acid tert-butyl ester (700 mg, 2.51 mmol) was dissolved in DCM (7 mL) at room temperature. Iodine monochloride (489 mg, 3.02 mmol) was added under ice bath condition. The reaction was stirred at room temperature for 1 hour under ice bath condition.

[0083] After the reaction was completed, water (5 mL) was added, and the mixture was extracted with ethyl acetate (15 mL x 3). The organic phase was combined and concentrated under reduced pressure. The residue was purified by silica gel column (DCM / MeOH = 10:1) to give (1S,3S)-3-((5-iodopyridin-2-yl-6-amino)cyclopentyl)carbamic acid tert-butyl ester as a yellow oily liquid (720 mg, 71% yield). LC-MS: [M+H]+ = 405.

[0084] Step D: Synthesis of (1S,3S)-N1-(5-iodopyridin-2-yl-6-d)cyclopentane-1,3-diamine hydrochloride

[0085]

[0086] (1S,3S)-3-((5-iodopyridin-2-yl-6-amino)cyclopentyl)carbamic acid tert-butyl ester (720 mg, 1.78 mmol) was dissolved in methanol (10 mL) at room temperature. Hydrochloric acid / dioxane (4.2 mL, 17.8 mmol) was added, and the reaction was stirred at room temperature for 2 hours.

[0087] ​The reaction was completed, concentrated to dryness to give 505 mg of (1S,3S)-N1-(5-iodopyridin-2-yl-6-d)cyclopentane-1,3-diamine hydrochloride as a light yellow solid (1S,3S)-N1-(5-iodopyridin-2-yl-6-d)cyclopentane-1,3-diamine hydrochloride was used directly in the next step without further purification. LC-MS: [M+H]+=305.

[0088] Step E: Synthesis of (1S,3S)-N1-(5-difluoromethoxy)pyrimidin-2-yl)-N3-(5-iodopyridin-2-yl-6-D)cyclopentane-1,3-diamine

[0089]

[0090] (1S,3S)-N1-(5-difluoromethoxy)pyrimidin-2-yl)-N3-(5-iodopyridin-2-yl-6-D)cyclopentane-1,3-diamine (500 mg, 1.12 mmol) was dissolved in DMSO (5 mL) at room temperature, pyridin-2(lH)-one (212 mg, 2.23 mmol), 8-hydroxyquinoline (49 mg, 0.335 mmol), cuprous iodide (64 mg, 0.335 mmol), potassium phosphate (509 mg, 3.68 mmol) were added, the reaction system was slowly warmed to 140 °C under nitrogen protection, and the reaction was carried out overnight.

[0091] The reaction was completed, water (20 ml) was added, and extraction was performed with ethyl acetate (60 ml x 3). The organic phases were combined and concentrated under reduced pressure, and the residue was purified by silica gel column (DCM / MeOH = 10:1) to give 500 mg of (1S,3S)-N1-(5-difluoromethoxy)pyrimidin-2-yl)-N3-(5-iodopyridin-2-yl-6-D)cyclopentane-1,3-diamine as a light yellow oily liquid (yield 75%). LC-MS: [M+H]+=449.

[0092] Step F: Synthesis of

[0093] 6'-(((1S,3S)-3-((5-(difluoromethoxy)pyrimidin-2-yl)amino)cyclopentyl)amino)-2H-[l,3'-bipyridinyl]-2-one-2'-d

[0094]

[0095] (1S,3S)-N1-(5-difluoromethoxy)pyrimidin-2-yl)-N3-(5-iodopyridin-2-yl-6-D)cyclopentane-1,3-diamine (500 mg, 1.12 mmol) was dissolved in DMSO (5 mL) at room temperature, pyridin-2(lH)-one (212 mg, 2.23 mmol), 8-hydroxyquinoline (49 mg, 0.335 mmol), cuprous iodide (64 mg, 0.335 mmol), potassium phosphate (509 mg, 3.68 mmol) were added, the reaction system was slowly warmed to 140 °C under nitrogen protection, and the reaction was carried out overnight.

[0096] The reaction was completed, water (10 ml) was added, and extraction was performed with ethyl acetate (80 ml x 3). The organic phase was combined, concentrated under reduced pressure, and the residue was purified by reverse phase column chromatography and lyophilized to obtain 210 mg of yellow solid product 6'-(((1S,3S)-3-((5-(difluoromethoxy)pyrimidin-2-yl)amino)cyclopentyl)amino)-2H-[1,3'-bipyridinyl]-2-one-2'-d (yield 44%).

[0097] LC-MS: [M+H]+=416.

[0098] 1H NMR (400 MHz, DMSO-d6) δ 8.23 (s, 2H), 7.64 - 7.56 (m, 1H), 7.47 (ddd, J = 8.8, 5.3, 2.1 Hz, 2H), 7.39 (d, J = 8.9 Hz, 1H), 7.24 - 6.80 (m, 2H), 6.53 (d, J = 8.9 Hz, 1H), 6.44 (d, J = 9.1 Hz, 1H), 6.27 (td, J = 6.7, 1.3 Hz, 1H), 4.37 - 4.24 (m, 2H), 2.19 - 2.04 (m, 2H), 1.94 - 1.80 (m, 2H), 1.58 - 1.40 (m, 2H).

[0099] Example 3

[0100] Synthesis of 6'-(((1S,3S)-3-((5-(difluoromethoxy)pyrimidin-2-yl-4-d)amino)cyclopentyl)amino)-2H-[1,3'-bipyridinyl]-2-one

[0101]

[0102] Synthesis route:

[0103]

[0104] Step A: Synthesis of 2-chloropyrimidin-4-deuterium-5-ol

[0105]

[0106] 2,4-dichloropyrimidin-5-ol (500 mg, 3.03 mmol) was added to a reaction bottle at room temperature, then deuterium water (10 ml) and zinc powder (500 mg, 9.09 mmol) were added, and stirring was performed at 100°C for 3 hours.

[0107] After the reaction was completed, it was poured into 50 mL of water, extracted twice with ethyl acetate (100 mL), the organic phase was discarded, the aqueous phase was adjusted to pH 3-4 with saturated citric acid solution, extracted with ethyl acetate (50 mL x 3), the combined organic phase was washed with saturated sodium chloride (50 mL), dried over anhydrous sodium sulfate, filtered, and concentrated to give 150 mg of yellow solid 2-chloropyrimidin-4-d-5-ol (yield: 37.8%). LC-MS: [M+H] + = 132.1.

[0108] Step B: Synthesis of 2-chloro-5-(difluoromethoxy)pyrimidin-d

[0109]

[0110] After the reaction was completed, it was poured into 50 mL of water, extracted twice with ethyl acetate (100 mL), the organic phase was discarded, the aqueous phase was adjusted to pH 3-4 with saturated citric acid solution, extracted with ethyl acetate (50 mL x 3), the combined organic phase was washed with saturated sodium chloride (50 mL), dried over anhydrous sodium sulfate, filtered, and concentrated to give 150 mg of yellow solid 2-chloropyrimidin-4-d-5-ol (yield: 37.8%). LC-MS: [M+H]

[0111] After the reaction was completed, it was poured into 50 mL of water, extracted twice with ethyl acetate (100 mL), the organic phase was discarded, the aqueous phase was adjusted to pH 3-4 with saturated citric acid solution, extracted with ethyl acetate (50 mL x 3), the combined organic phase was washed with saturated sodium chloride (50 mL), dried over anhydrous sodium sulfate, filtered, and concentrated to give 150 mg of yellow solid 2-chloropyrimidin-4-d-5-ol (yield: 37.8%). LC-MS: [M+H]

[0112] Step C: Synthesis of (6'-(((1S,3S)-3-((5-(difluoromethoxy)pyrimidin-2-yl-4-d)amino)cyclopentyl)amino)-2H-[1,3'-bipyridin]-2-one

[0113]

[0114] After the reaction was completed, it was poured into 50 mL of water, extracted twice with ethyl acetate (100 mL), the organic phase was discarded, the aqueous phase was adjusted to pH 3-4 with saturated citric acid solution, extracted with ethyl acetate (50 mL x 3), the combined organic phase was washed with saturated sodium chloride (50 mL), dried over anhydrous sodium sulfate, filtered, and concentrated to give 150 mg of yellow solid 2-chloropyrimidin-4-d-5-ol (yield: 37.8%). LC-MS: [M+H]

[0115] After the reaction was completed, it was poured into 50 mL of water, extracted twice with ethyl acetate (100 mL), the organic phase was discarded, the aqueous phase was adjusted to pH 3-4 with saturated citric acid solution, extracted with ethyl acetate (50 mL x 3), the combined organic phase was washed with saturated sodium chloride (50 mL), dried over anhydrous sodium sulfate, filtered, and concentrated to give 150 mg of yellow solid 2-chloropyrimidin-4-d-5-ol (yield: 37.8%). LC-MS: [M+H]

[0116] LC-MS: [M+1]+= 416.2.

[0117] 1H NMR (400 MHz, DMSO-d6) δ 8.25 (s, 1H), 7.92 (d, J = 2.8 Hz, 1H), 7.61 (dd, J = 6.8, 2.0 Hz, 1H), 7.54 - 7.44 (m, 2H), 7.40 (dd, J = 8.8, 2.8 Hz, 1H), 7.24 - 6.82 (m, 2H), 6.55 (d, J = 8.0 Hz, 1H), 6.45 (d, J = 12 Hz, 1H), 6.27 (td, J = 6.8, 1.2 Hz, 1H), 4.40 - 4.25 (m, 2H), 2.20 - 2.06 (m, 2H), 1.97 - 1.80 (m, 2H), 1.61 - 1.44 (m, 2H).

[0118] Example 4

[0119] Synthesis of 6'-(((1S,3S)-3-((5-(difluoromethoxy)pyrimidin-2-yl-4,6-d2)amino)cyclopentyl)amino)-2H-[1,3'-bipyridinyl]-2-one

[0120]

[0121] Synthesis route:

[0122]

[0123] Step A: Synthesis of 2-chloropyrimidine-4,6-d2-5-ol

[0124]

[0125] 2,4,6-trichloropyrimidine-5-ol (500 mg, 3.03 mmol) was added to a reaction flask followed by deuterium water (10 mL) and zinc powder (500 mg, 9.09 mmol) and stirred at 100 °C for 3 h.

[0126] After completion of the reaction, it was poured into 50 mL of water and extracted with ethyl acetate (100 mL) twice. The organic phase was discarded and the aqueous phase was adjusted to pH 3-4 with saturated citric acid solution and extracted with ethyl acetate (50 mL x 3). The combined organic phase was washed with saturated sodium chloride (50 mL), dried over anhydrous sodium sulfate, filtered and concentrated to obtain 100 mg of 2-chloropyrimidine-4,6-d2-5-ol as a yellow solid (yield: 30.2%). LC-MS: [M+H]+= 133.1.

[0127] Step B: Synthesis of 2-chloro-5-(difluoromethoxy)pyrimidine-4,6-d2

[0128]

[0129] To a reaction flask was added 2-chloropyrimidine-4,6-d2-5-ol (100 mg, 0.75 mmol), potassium carbonate (313 mg, 2.27 mmol), N,N-dimethylformamide (2 mL), sodium difluorochloroacetate (345 mg, 2.27 mmol) at room temperature and stirred at 100 °C for 3 h.

[0130] The reaction was complete, poured into 20 mL of water, extracted with ethyl acetate (20 mL) twice, combined organic phase washed with saturated sodium chloride (20 mL), dried over anhydrous sodium sulfate, filtered, concentrated to give 65 mg of 2-chloro-5-(difluoromethoxy)pyrimidine-4,6-d2 as brown oil (yield: 47.5%).

[0131] Step C: 6'-(((lS,3S)-3-((5-(difluoromethoxy)pyrimidin-2-yl-4,6-d2)amino)cyclopentyl)amino)- 2H-[l,3'-bipyridinyl]-2-one

[0132]

[0133] To a reaction flask was added 2-chloro-5-(difluoromethoxy)pyrimidine-4,6-d2 (65 mg, 0.357 mmol), dimethyl sulfoxide (2 mL) and 6'-(((lS,3S)-3-aminocyclopentyl)amino)-2H- [l,3'-bipyridinyl]-2-one hydrochloride (116 mg, 0.428 mmol), N,N-diisopropylethylamine (0.25 mL) at room temperature and stirred at 110 °C overnight.

[0134] The reaction was complete, filtered, the filtrate was purified by reverse phase column (acetonitrile / water = 5% to 65%) to give 35 mg of 6'-(((lS,3S)-3-((5-(difluoromethoxy)pyrimidin-2-yl-4,6-d2)amino)cyclopentyl)amino)-2H- [l,3'-bipyridinyl]-2-one as yellow solid (yield: 24.0%). LC-MS: [M+l] = 417.3. - = 417.3. 1H NMR (400 MHz, DMSO-d6) δ 7.92 (d, J = 2.8 Hz, 1H), 7.60 (dd, J = 6.8, 2.0 Hz, 1H), 7.50 - 7.46 (m, 2H), 7.40 (dd, J = 8.8, 2.8 Hz, 1H), 7.24 - 6.81 (m, 2H), 6.54 - 6.43 (m, 2H), 6.27 (td, J = 6.8, 1.2 Hz, 1H), 4.31 (q, J = 6.8, 6.4 Hz, 2H), 2.25 - 2.03 (m, 2H), 1.99 - 1.74 (m, 2H), 1.59 - 1.42 (m, 2H).

[0135] Example 5 Related Activity Testing

[0136] Test Method:

[0137] Biacore TMSurface Plasmon Resonance data were collected on a 8K system (GE Healthcare). Streptavidin was immobilized on a SA (Cytiva) sensor chip using standard amine coupling chemistry with NBS-N (10 mM HEPES, 0.15 M NaCl, Ph 7.4) as running buffer at 25°C. Briefly, the carboxymethyl dextran surface was activated by a 12 min injection of 0.4 M 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide hydrochloride (EDC) / 0.1 M N-hydroxysuccinimide (NHS) at a flow rate of 10 μl / min in a 1:1 ratio. For capture of streptavidin, the protein was diluted to 0.5 mg / ml in 10 mM sodium acetate (Ph 4.5) and captured by injecting 100 ul to the active chip surface. Excess of residual activated groups were blocked by a 7 min injection of 1 M ethanolamine (pH 8.5). 5 ul of protein diluted to 10 μg / mL in NBS-N, 0.05% Tween-20, 0.1 mM CaCl2 at a flow rate of 5 μl / min for 60 s. Aci-tagged PCSK9 protein was captured on the streptavidin surface. Typical surface densities obtained were 2900-3200 RU. SPR binding data were obtained using appropriate dilution series of each compound at a flow rate of 30 μl / min, capture time of 60 s / concentration point and dissociation time of 3600 s. The running buffer used for compound binding studies was 10 mM HEPES, Ph 7.4, 150 mM NaCl, 0.05% P20, 2% DMSO. Data were corrected for DMSO and did not include volume effects. All data were double referenced against a blank injection and a reference surface and data processing and kinetic fitting were performed using Scrubber software version 2.0c (Biological Software). Data were fitted using a simple 1:1 binding model to determine K D values.

[0138] Test results:

[0139] K D The results of the test are shown in Table 1 below:

[0140] Table 1. PCSK9 Kinetics data

[0141]

[0142] From the above results, it can be seen that the activity of the deuterated compound of the present application is slightly better than that of the non-deuterated compound.

[0143] Effect of the compound of Example 6 on hERG current in hERG-HEK293 cells

[0144] Test method:

[0145] hERG-HEK293 cells stably expressing hERG channels were chosen for the experiment. The whole-cell voltage-clamp mode was formed by clamping hERG-HEK293 cells with a full-automatic patch-clamp system, and the hERG current was induced by the corresponding voltage. The cells were given 30 μM of the compound containing 0.3% DMSO in the extracellular fluid (negative control) respectively. Different cells were given 1, 10, 100, and 1000 nM of the positive control (cisapride) containing 0.3% DMSO in the extracellular fluid (negative control) respectively. The hERG channel tail current was recorded, and the peak tail current at each concentration was obtained. The inhibition rate of the current after adding the compound and the positive control was calculated. The concentration-response relationship curve fitting and IC50 calculation of the positive control were completed with the GraphPad Prism software.

[0146] Table 2

[0147] Example 30 μM - % inhibition 1 68.51±1.08 2 40.76±3.33 Control compound 51.47±1.36

[0148] From the above results, it can be seen that the hERG risk of compound 2 is smaller.

[0149] Example 7 Compound monkey liver microsome study

[0150] (1) Experimental materials

[0151] The monkey liver microsomes were purchased from Red Liver Disease Research (Shanghai) Co., Ltd.

[0152] Reagents: DMSO (dimethyl sulfoxide), acetonitrile, formic acid, propranolol (internal standard) were all commercially available.

[0153] Instrument: Thermo LC-MS (U300 UPLC, TSQ QUANTUMN ULTRA triple quadrupole mass spectrometer).

[0154] (2) Experimental method

[0155] A certain amount of compound was precisely weighed into DMSO to make a 10 mM stock solution, which was diluted with diluent (ACN:H2O = 1:1) to 100 μM working solution, and then with 0.1 M potassium phosphate buffer to 3 μM dosing solution for standby. 75 μL of liver microsomes was added to 925 μL of 0.1 M potassium phosphate buffer to mix evenly to make 1.5 mg / mL liver microsomal suspension, which was pre-incubated at 37°C for 10 min. Preparation at 0 point: 15 μL of the above liver microsomal suspension was added to 6 mM NADPH solution, and then 150 μL of propranolol acetonitrile solution was added immediately to precipitate, and then 15 μL of the above dosing solution was added to mix evenly for standby. Preparation of 20 min and 60 min samples: 15 μL of the dosing solution was added to 15 μL of the liver microsomal suspension and 15 μL of 6 mM NADPH solution to mix evenly, and then incubated at 37°C for 20 min and 60 min, respectively. The above sample preparation was double-well parallel operation. When the above samples were incubated to the relevant time points, 150 μL of propranolol acetonitrile solution was added to terminate the reaction. All the above samples were centrifuged at 4000 rpm for 5 min, and then 100 μL of supernatant was added to 100 μL of ultrapure water to mix evenly before LC-MS / MS analysis. The LC-MS / MS detection conditions are as follows:

[0156] Chromatographic column: Waters ACQUITY™ PREMIER HSS T3, 50*2.1 mm, 1.8 μm.

[0157] Mobile phase: water (0.1% formic acid) - acetonitrile gradient elution according to the following table

[0158] Table 3

[0159] Time (min) Water (containing 0.1% formic acid) Acetonitrile 0 85％ 15％ 0.6 85％ 15％ 1 20％ 80％ 2.3 20％ 80％ 2.31 85％ 15％ 3 85％ 15％

[0160] (3) Data processing

[0161] The results are shown in Table 4:

[0162] Conc. (μM) ％ / 2 Verapmil 1 2.2 3 1 764.5 4 1 800.4 Control compound 1 407.4

[0163] From the above data, it can be seen that deuterated compounds 3 and 4 are more stable than non-deuterated control compound monkey liver microsomes.

[0164] Example 8 in vivo stability experiment

[0165] 1. Reagents and instruments

[0166] PEG-400 (batch number R22040588, Shanghai Shaoyuan Reagent Co., Ltd.), DMSO (batch number 20200319, Guangdong Guanghua Technology Co., Ltd.), normal saline (batch number 2011110727, Chenxin Pharmaceutical Co., Ltd.). LC-MS instrument (Thermo Fisher Ultimate 3000 UPLC, TSQ QUANTUM ULTRA triple quadrupole mass spectrometer, AB SCIEX 5500+ QTARP).

[0167] 2. Experimental animals

[0168] SD rats: male, 180-250 g, purchased from Guangdong Weitong Lihua Experimental Animal Technology Co., Ltd.

[0169] 3. Preparation of formulations

[0170] The test product powder was accurately weighed, completely dissolved in DMSO, then PEG-400 was added, vortexed and ultrasonically mixed, then normal saline was added, vortexed and ultrasonically mixed to form 0.5 mg / mL (DMSO: PEG-400: NS = 5:60:35, V / V / V), 10 mL / kg for oral administration, 2 mL / kg for intravenous administration.

[0171] 4. Blood sample collection

[0172] After intravenous or oral administration in rats, 200 μL of venous blood was collected at 5 min (not for oral administration), 15 min, 30 min, 1 h, 2 h, 5 h, 7 h, and 24 h into an EDTA-K2-anticoagulant EP tube, centrifuged at 10,000 rpm for 2 min, and the plasma was stored at -80°C for testing.

[0173] 5. Biological analysis

[0174] A certain amount of test product was accurately weighed and dissolved in DMSO to 2 mg / mL as a stock solution. An appropriate amount of compound stock solution was accurately pipetted, diluted with acetonitrile to prepare a standard series of solutions. 4 μL of each of the above standard series of solutions was accurately pipetted, 36 μL of blank plasma was added, vortexed to mix, and plasma samples corresponding to concentrations of 1, 3, 5, 10, 30, 100, 300, 1000, and 3000 ng / mL were prepared. Each concentration was analyzed in duplicate to establish a standard curve. 30 μL of plasma was taken, 200 μL of propranolol (5 ng / mL) in acetonitrile was added, vortexed to mix, then centrifuged at 4000 rpm for 10 min, and the supernatant was analyzed by LC-MS. The LC-MS detection conditions are as follows:

[0175] Chromatographic column: YMC-Triart C18, 50x2.1mm, S-3μm 12nm.

[0176] Mobile phase A: water (0.1% formic acid), mobile phase B: acetonitrile, flow rate: 0.5 mL / min, gradient elution as shown in Table 5 below:

[0177] Table 5

[0178] Time (min) A(％) B(％) 0 80％ 20％ 1.2 20％ 80％ 2.6 20％ 80％ 2.61 80％ 20％ 3.0 80％ 20％

[0179] 6. Data processing

[0180] After LC-MS detection of blood drug concentration, WinNonlin 6.1 software was used to calculate the pharmacokinetic parameters of rats after administration by non-compartment model, and the results are shown in Table 6 below.

[0181] Table 6: Rat pharmacokinetic parameters of compounds of the present application (iV and PO administration)

[0182]

[0183]

[0184] From the above data, it can be seen that deuterated compounds 2 and 3 have longer half-lives and slower clearance rates relative to non-deuterated control compounds.

[0185] Example 9: Test of binding affinity of compound of Example 9 to PCSK9 protein

[0186] The compound of the present application is used to determine the binding affinity of the compound to PCSK9 protein by fluorescence polarization method.

[0187] All compounds were dissolved in DMSO to prepare 10 mM stock solution. The positive compound and the test compound were diluted with DMSO in 5-fold gradient from 10 mM, a total of 8 concentration gradients. First, a certain volume of fluorescent probe solution was prepared using test buffer (20 mM HEPES, 150 mM NaCl, 1 mM CaCl2 and 0.01% Tween-20) to make the concentration 5 nM; then the series of DMSO solutions of test compounds were diluted 50 times with the fluorescent probe solution; finally, the human recombinant PCSK9 protein (provided by Chengdu Xintai) solution with a concentration of 9 μg / mL was prepared with the test buffer. After the test solution was prepared, 7.5 μL of PCSK9 protein was added to a black 384-well plate (PerkinElmer, Cat#6008260), and 7.5 μL of compound and fluorescent probe mixed solution with different concentrations (DMSO final concentration was 1%) was added, while setting up a positive control group (test buffer + equal volume of target protein + equal proportion of fluorescent probe molecule) and a negative control group (test buffer + equal proportion of fluorescent probe molecule), the final concentration of the probe molecule in the system was 2.5 nM, and the concentration of PCSK9 protein was 4.5 μg / mL. After incubation at room temperature for 15 minutes, the fluorescence polarization value was read using an enzyme-labeled instrument at an excitation wavelength of 485 nm and an emission wavelength of 520 nm. The inhibition rate of the drug = [1-(mP( 药物筛选组 )-mP( 阴性对照组 )] ÷ [mP( 阳性对照组 )-mP( 阴性对照组 ))] x 100. The logarithmic value of the concentration of the compound was taken as the abscissa, and the inhibition rate was taken as the ordinate. The 4-parameter nonlinear regression curve fitting was carried out to calculate the IC50 value (Y = Bottom + (Top-Bottom) / (1+10^((LogIC 50 -X)*HillSlope)), wherein: Hillslope represents the slope of the curve, and IC 50 represents the half-inhibitory concentration.

[0188] Table 7. FP-IC 50 Data

[0189] Example FP-IC 50 (nM) 2 65.21 3 75.46 4 69.71 Control compound 121.29

[0190] From the above results, it can be seen that the binding affinity of the deuterated compound PCSK9 protein of the present application is slightly better than that of the non-deuterated compound.

[0191] The structural formulas of the control compounds in Examples 5, 6, 7, 8 and 9 are as follows:

[0192]

[0193] The above embodiments are the preferred embodiments of the present application, but the embodiments of the present application are not limited to the above embodiments, and any changes, modifications, substitutions, combinations, simplifications, etc. made without departing from the spirit and principles of the present application should be equivalent replacement manners and should be included in the protection scope of the present application.

Claims

1. A compound, or a racemate thereof, or a pharmaceutically acceptable salt thereof, characterized in that, selected from the group consisting of:

2. The compound according to claim 1, or a racemate thereof, or a pharmaceutically acceptable salt thereof, wherein, The pharmaceutically acceptable salt refers to the compound, or its racemate, prepared with a pharmaceutically acceptable acid or base.

3. A pharmaceutical composition, characterized in that A pharmaceutical composition comprising a therapeutically effective amount of the compound of claim 1, or its racemate, or its pharmaceutically acceptable salt, and a pharmaceutically acceptable carrier.

4. Use of the compound of claim 1, or its racemate, or its pharmaceutically acceptable salt, in the manufacture of a medicament for treating a disease, wherein the disease is a PCSK9 inhibitor related disease.

5. The medical use according to claim 4, wherein the PCSK9 inhibitor related disease is selected from the group consisting of hypercholesterolemia.

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