Anthracycline compound, preparation method therefor, intermediate, and use thereof

Abstract

Disclosed are an anthracycline compound, a preparation method therefor, an intermediate, and a use thereof. The present invention provides an anthracycline compound as shown in formula I or a pharmaceutically acceptable salt thereof. The anthracycline compound provided by the present invention has antitumor activity comparable to existing anthracycline drugs (such as nemorubicin) or greater than that of existing anthracycline drugs doxorubicin and daunorubicin, and does not form an oxazole ring structure during metabolism, thereby improving the clinical safety of such drugs.

Description

An anthracycline compound, preparation method, intermediates and applications thereof

[0001] This application claims priority to Chinese Patent Application No. 2024110516049, filed on August 1, 2024. This application incorporates the entirety of the aforementioned Chinese Patent Application. TECHNICAL FIELD

[0002] The present application relates to the field of organic compounds, in particular to an anthracycline compound, preparation method, intermediates and applications thereof, which can be used for the treatment of tumors. BACKGROUND

[0003] Anthracyclines are a class of natural antibiotics, which are one of the most effective anticancer drugs in clinical use. They are linked by a glycosidic bond between an aminosugar structure and an anthracycline. This class of anthracyclines acts through DNA insertion, oxidative stress and topoisomerase II poisoning. Despite their significant clinical tumor treatment effect, multidrug resistance and severe cardiotoxicity remain the most important limitations of this class of anthracyclines, prompting people to discover new analogs to circumvent this drawback.

[0004] Doxorubicin (adriamycin) is one of the most commonly used and effective chemotherapy drugs in clinical practice. It is used as a first-line treatment for various cancers, including lymphoma, sarcoma and various solid tumors, such as breast cancer, lung cancer, bladder cancer, bone cancer and cervical cancer in children and adults. Due to the relatively weak cell inhibitory activity of doxorubicin, the amount of drug needed to be increased in clinical practice, which can lead to dose-related cardiotoxicity. Although the advent of doxorubicin liposome, which can reduce the exposure of doxorubicin in the blood while increasing the amount of drug, has reduced cardiotoxicity, the large amount of drug used is still a hidden danger for drug safety.

[0005] Daunorubicin is an anthracycline drug used to treat acute lymphoblastic and myeloblastic leukemia. Due to the anthracycline parent nucleus structure, it can cause cardiotoxicity in clinical use, and there is also multidrug resistance. Low-dose long-term administration is often used in clinical practice to reduce cardiotoxicity.

[0006] In order to solve the problem of cardiotoxicity, many studies have been conducted to modify the anthracycline structure. Nemozolubicin (morpholino anthracycline toxin) is a drug modified based on the structure of doxorubicin. Its antitumor activity is nearly 100 times that of doxorubicin and daunorubicin, so it is expected to reduce cardiotoxicity by reducing the amount of drug used. However, clinical studies have found that nemozolubicin produces a primary metabolite PNU-159682 after being metabolized by the liver, which has an activity of more than 1000 times that of nemozolubicin. Therefore, the drug is limited in further research due to its narrow dosing window. The structures of nemozolubicin and PNU-159682 are as follows:

[0007] The high cytotoxicity of PNU-159682 is related to its unique derivative structure, so further modification of the morpholino anthracycline toxin to avoid the formation of oxazolidine ring structure in the metabolic process and to avoid the generation of metabolites similar to the structure of PNU-159682 (doi: 10.1158 / 1078-0432.CCR-04-1845) is expected to improve the clinical safety of this class of drugs. SUMMARY

[0008] The technical problem to be solved by the present application is to avoid the generation of metabolites containing oxazole ring structure in the metabolic process of drugs, such as PNU-159682; To this end, the present application provides an anthracycline compound and a preparation method, intermediates and applications thereof. The anthracycline compound provided by the present application has an antitumor activity comparable to or higher than that of existing anthracycline drugs (such as nemorubicin) and doxorubicin and daunorubicin, and will not form an oxazole ring structure in the metabolic process, thereby improving the clinical safety of this class of drugs.

[0009] The present application provides an anthracycline compound as shown in formula I or a pharmaceutically acceptable salt thereof:

[0010] R 1 and R 2 are independently H, F, deuterium, C 1-6 alkyl, or R 1 , R 2 and the C atom connected thereto together form a cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl group; and R 1 and R 2 are not simultaneously H;

[0011] R 3 and R 4 are independently H, F, deuterium, C 1-6 alkyl, or R 3 , R 4 and the C atom connected thereto together form a cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl group; and R 3 and R 4 are not simultaneously H;

[0012] R 5 is hydrogen, C 1-6 alkyl, cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl.

[0013] In a preferred embodiment, certain substituents in the compounds of Formula I, or pharmaceutically acceptable salts thereof, can further have the following definitions, and the definitions of substituents not mentioned below are as described in any of the embodiments of the present application (hereinafter referred to as "in a preferred embodiment").

[0014] In a preferred embodiment, R 1 , R 2 , R 3 , and R 4 , the C 1-6 alkyl is independently methyl, ethyl, n-propyl, i-propyl, n-butyl, s-butyl, i-butyl, or t-butyl; for example, methyl or ethyl; and further for example, methyl.

[0015] In a preferred embodiment, R 1 and R 2 , R 1 , R 2 together with the C atom to which they are attached form a cyclopropyl or cyclobutyl group; for example, cyclopropyl.

[0016] In a preferred embodiment, R 3 and R 4 , R 3 , R 4 together with the C atom to which they are attached form a cyclopropyl or cyclobutyl group; for example, cyclopropyl.

[0017] In a preferred embodiment, R 5 , the C 1-6 alkyl is independently methyl, ethyl, n-propyl, i-propyl, n-butyl, s-butyl, i-butyl, or t-butyl; for example, methyl or ethyl; and further for example, methyl.

[0018] In a preferred embodiment, R 1 and R 2 are independently F, deuterium, C 1-6 alkyl, or R 1 , R 2 together with the C atom to which they are attached form a cyclopropyl, cyclobutyl, cyclopentyl, or cyclohexyl group.

[0019] R 3 and R 4 are independently F, deuterium, C 1-6 alkyl, or R 3 , R 4 together with the C atom to which they are attached form a cyclopropyl, cyclobutyl, cyclopentyl, or cyclohexyl group.

[0020] In a preferred embodiment, R 1 and R 2 are independently C 1-6 alkyl, or R 1 , R2 with the C atom to which they are attached together form cyclopropyl or cyclobutyl; preferably, R 1 and R 2 are independently methyl, or R 1 , R 2 and the C atom to which they are attached together form cyclopropyl.

[0021] In a certain preferred embodiment, R 1 is C 1-6 alkyl.

[0022] In a certain preferred embodiment, R 1 is H or deuterium.

[0023] In a certain preferred embodiment, R 1 is H, ethyl, F or deuterium.

[0024] In a certain preferred embodiment, R 1 is H, methyl or deuterium.

[0025] In a certain preferred embodiment, R 2 is F, deuterium or C 1-6 alkyl.

[0026] In a certain preferred embodiment, R 2 is deuterium or C 1-6 alkyl.

[0027] In a certain preferred embodiment, R 2 is C 1-6 alkyl.

[0028] In a certain preferred embodiment, R 2 is deuterium.

[0029] In a certain preferred embodiment, R 2 is ethyl, F or deuterium.

[0030] In a certain preferred embodiment, R 2 is methyl or deuterium.

[0031] In a certain preferred embodiment, R 3 and R 4 are independently C 1-6 alkyl, or R 3 , R 4 and the C atom to which they are attached together form cyclopropyl or cyclobutyl; preferably, R 3 and R 4 are independently methyl, or R 3 , R 4 and the C atom to which they are attached together form cyclopropyl.

[0032] In a certain preferred embodiment, R 3 is C1-6 alkyl.

[0033] In a certain preferred solution, R 3 It can be H or deuterium.

[0034] In a certain preferred solution, R 3 It can be H, ethyl, F or deuterium.

[0035] In a certain preferred solution, R 3 It can be H, methyl, or deuterium.

[0036] In a certain preferred solution, R 4 For F, deuterium or C 1-6 alkyl.

[0037] In a certain preferred solution, R 4 For deuterium or C 1-6 alkyl.

[0038] In a certain preferred solution, R 4 It is deuterium.

[0039] In a certain preferred solution, R 4 C 1-6 alkyl.

[0040] In a certain preferred solution, R 4 It can be ethyl, F, or deuterium.

[0041] In a certain preferred solution, R 4 It is methyl or deuterium.

[0042] In a certain preferred solution, R 1 and R 2 same.

[0043] In a certain preferred solution, R 3 and R 4 same.

[0044] In a certain preferred solution, R 1 R 2 R 3 and R 4 same.

[0045] In a certain preferred solution, R 5 C 1-6 Alkyl, cyclopropyl, or cyclobutyl; preferably, R 5 C 1-6 Alkyl groups, such as methyl groups.

[0046] In a preferred embodiment, the anthracene ring compound as shown in Formula I is an anthracene ring compound as shown in Formula IX.

[0047] R 5 is C 1-6 alkyl, cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl.

[0048] In a certain preferred embodiment, the anthracycline compound of formula I is an anthracycline compound of formula I-Y,

[0049] R 5 is C 1-6 alkyl, cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl.

[0050] In a certain preferred embodiment, in the anthracycline compound of formula I,

[0051] R 1 and R 2 are independently C 1-6 alkyl, or R 1 , R 2 and the C atom to which they are attached together form a cyclopropyl or cyclobutyl group;

[0052] R 3 and R 4 are independently C 1-6 alkyl, or R 3 , R 4 and the C atom to which they are attached together form a cyclopropyl or cyclobutyl group;

[0053] R 5 is C 1-6 alkyl.

[0054] In a certain preferred embodiment, R 1 and R 2 are independently C 1-6 alkyl, R 3 and R 4 are independently C 1-6 alkyl, R 5 is C 1-6 alkyl.

[0055] In a certain preferred embodiment, R 1 is H or deuterium, R 2 is deuterium; R 3 is H or deuterium, R 4 is deuterium; R 5 is C 1-6 alkyl.

[0056] In a certain preferred embodiment, the anthracycline compound of formula I is any one of the following compounds:

[0057] The present application provides a method for preparing an anthracycline compound of Formula I, comprising the step of: subjecting a compound of Formula I-1 to a deprotection reaction as shown in the following formula in the presence of a deprotection reagent (e.g., tetrabutylammonium fluoride) in a solvent (e.g., tetrahydrofuran) to obtain an anthracycline compound of Formula I,

[0058] R 1 , R 2 , R 3 , R 4 , and R 5 are as described in any of the Schemes, R a and R b are independently a hydroxyl protecting group, for example R a is a tert-butyldiphenylsilyl group (TBDPS-), R b is an acetyl group.

[0059] In a certain preferred embodiment, the compound of Formula I-1 is a compound of Formula I-1-X

[0060] The anthracycline compound of Formula I is a compound of Formula I-X.

[0061] In a certain preferred embodiment, the compound of Formula I-1 is a compound of Formula I-1-Y

[0062] The anthracycline compound of Formula I is a compound of Formula I-Y.

[0063] In a certain preferred embodiment, the method for preparing an anthracycline compound of Formula I further comprises a method for preparing a compound of Formula I-1, the method for preparing a compound of Formula I-1 comprising the step of: subjecting a compound of Formula I-A and a compound of Formula I-B to an addition reaction as shown in the following formula in the presence of an acid (e.g., p-toluenesulfonic acid) in a solvent (e.g., toluene) to obtain a compound of Formula I-1,

[0064] R 1 , R 2 , R 3 , R 4 , R 5 , R a , and R b are as described in any of the Schemes.

[0065] In a certain preferred embodiment, the compound of Formula I-A is

[0066] In a certain preferred embodiment, the compound of Formula I-A is

[0067] In a preferred embodiment, the process for the preparation of an anthracene derivative of formula I further comprises a process for the preparation of a compound of formula I-1-X, said process for the preparation of a compound of formula I-1-X comprising the step of performing an addition reaction of a compound of formula I-A-X and a compound of formula I-B in a solvent (e.g. toluene) in the presence of an acid (e.g. p-toluenesulfonic acid) to give a compound of formula I-1-X,

[0068] R 1 , R 2 , R 3 , R 4 , R 5 , R a and R b are defined as described in any one embodiment of the present application.

[0069] The present application provides a process for the preparation of a compound of formula I-1, comprising the step of performing an addition reaction of a compound of formula I-A and a compound of formula I-B in a solvent (e.g. toluene) in the presence of an acid (e.g. p-toluenesulfonic acid) to give a compound of formula I-1,

[0070] R 1 , R 2 , R 3 , R 4 , R 5 , R a and R b are defined as described in any one embodiment.

[0071] The present application provides a process for the preparation of a compound of formula I-1-X, comprising the step of performing an addition reaction of a compound of formula I-A-X and a compound of formula I-B in a solvent (e.g. toluene) in the presence of an acid (e.g. p-toluenesulfonic acid) to give a compound of formula I-1-X,

[0072] R 1 , R 2 , R 3 , R 4 , R 5 , R a and R b are defined as described in any one embodiment.

[0073] The present application provides a process for the preparation of a compound of formula I-1,

[0074] R 1 , R 2 , R 3 , R 4 , R 5 , R a and R b are as defined in any of the aspects.

[0075] In a certain preferred aspect, the compound as shown in Formula I-1 is

[0076] In a certain preferred aspect, the compound as shown in Formula I-1 is any one of the following compounds:

[0077] The present application provides a compound as shown in Formula I-A,

[0078] R 1 , R 2 , R 3 , R 4 , R 5 , R a and R b are as defined in any of the aspects.

[0079] In a certain preferred aspect, the compound as shown in Formula I-A

[0080] In a certain preferred aspect, the compound as shown in Formula I-A is any one of the following compounds:

[0081] The present application provides a pharmaceutical composition comprising:

[0082] (1) (a prophylactically or therapeutically effective amount of) an anthracycline compound as shown in Formula I or a pharmaceutically acceptable salt thereof according to any aspect of the present application; and

[0083] (2) a pharmaceutical adjuvant.

[0084] The present application also provides use of an anthracycline compound as shown in Formula I or a pharmaceutically acceptable salt thereof according to any aspect or the pharmaceutical composition in the preparation of a medicament for preventing and / or treating a tumor.

[0085] In a certain aspect, the tumor is malignant lymphoma, non-small cell lung cancer, ovarian cancer, breast cancer or Kaposi's sarcoma.

[0086] In certain embodiments, the tumor is a malignant lymphoma, lung cancer, cervical cancer, ovarian cancer, breast cancer, or Kaposi's sarcoma.

[0087] In certain embodiments, the tumor is lung cancer or cervical cancer, which can be non-small cell lung cancer.

[0088] The term "pharmaceutically acceptable" means that the salt, solvent, adjuvant, etc. is not toxic or safe and suitable for use with patients.

[0089] The term "alkyl" means a straight or branched chain alkyl group having the number of carbon atoms specified. Examples of alkyl groups include methyl, ethyl, n-propyl, i-propyl, n-butyl, t-butyl, i-butyl, s-butyl, n-pentyl, n-hexyl, and the like.

[0090] The term "pharmaceutically acceptable adjuvant" means excipients and additives used in the production of pharmaceuticals and dispensing of prescriptions, and all substances included in the pharmaceutical formulation other than the active ingredient.

[0091] The term "treatment" refers to therapeutic treatment. In reference to a particular condition, treatment refers to: (1) alleviating one or more of the biological manifestations of the disease or condition, (2) interfering with (a) one or more points in the biological cascade leading to or causing the condition or (b) one or more of the biological manifestations of the condition, (3) ameliorating one or more symptoms, effects, or side effects associated with the condition, or one or more symptoms, effects, or side effects associated with the condition or its treatment, or (4) slowing the development of the condition or one or more of the biological manifestations of the condition.

[0092] The term "prevention" refers to a reduction in the risk of acquiring or developing a disease or disorder.

[0093] The term "therapeutically effective amount" refers to the amount of a compound that, when administered to a patient, is sufficient to effect treatment of a disease or condition as described herein. The "therapeutically effective amount" will vary depending on the compound, the condition and its severity, and the age of the patient to be treated, but can be adjusted by those skilled in the art as needed.

[0094] The term "patient" refers to any animal, preferably a mammal, and most preferably a human, to which a compound or composition according to embodiments of the present application is administered. The term "mammal" includes any mammal. Examples of mammals include, but are not limited to, cows, horses, sheep, pigs, cats, dogs, mice, rats, rabbits, guinea pigs, monkeys, humans, and the like, with humans being most preferred.

[0095] The above preferred conditions can be combined in any way, without departing from the scope of the present application, to provide preferred embodiments of the present application.

[0096] The reagents and materials used in the present application are commercially available.

[0097] The positive progress effect of the present application is that the anti-tumor activity of the compound of the present application is higher than that of doxorubicin and daunorubicin, and is equivalent to that of the prototype drug, nemonoxin, and the modification of the anthracycline compound does not excessively reduce its activity, so that the cardiotoxicity can be reduced by reducing the dosage.

[0098] In addition, through metabolism experiments, the compound of the present application does not form a metabolite with high toxicity similar to the structure of PNU-159682, and is expected to improve the safety of drug use. DETAILED DESCRIPTION

[0099] The present application will be further described below by way of examples, but the present application is not limited to the scope of the examples described. The experimental methods in the following examples are not specified, and are selected according to conventional methods and conditions, or according to the instructions of the commercial product.

[0100] The experimental methods in the following examples are not specified, and are selected according to conventional methods and conditions, or according to the instructions of the commercial product.

[0101] The instrument parameters involved in the examples of the present application are as follows:

[0102] TLC detector instrument model: Liaposi three-purpose ultraviolet analyzer (BOS-203).

[0103] Nuclear magnetic resonance hydrogen spectrum instrument model: Bruker AVNEO 600 nuclear magnetic resonance spectrometer. 1 H-NMR) instrument model: Bruker AVNEO 600 nuclear magnetic resonance spectrometer.

[0104] Solvent: deuterated dimethyl sulfoxide (DMSO-d6), reference standard (δDMSO = 2.50).

[0105] Silica gel chromatography column model: commercially available conventional glass silica gel column (capacity specification is selected according to the amount of purified product).

[0106] Silica gel: 200-300 mesh commercially available silica gel (Qingdao Marine).

[0107] Liquid chromatography-mass spectrometry: Thermo Q-Exactive HF type quadrupole-ultra-high electrostatic field orbitrap high-resolution mass spectrometry system, equipped with High Chem Mass Frontier 7.0 software, UltiMate3000 type ultra-high performance liquid chromatograph.

[0108] Synthesis of nemonoxin derivatives in example group 1

[0109] The nemonoxin analogs of the present application can be obtained from formula A and formula B by addition reaction to obtain formula C, and further by removing the protecting group to obtain, and the reaction general formula is as follows:

[0110] wherein R 5 independently is a hydrogen, C 1-6 alkyl, cyclopropyl or cyclobutyl group;

[0111] R 1 and R 2 independently is a F atom, a deuterium atom, C 1-6 alkyl, or R 1 , R 2 and the C atom to which they are attached together form a cyclopropyl or cyclobutyl group;

[0112] R 3 and R 4 independently is a F atom, a deuterium atom, C 1-6 alkyl, or R 3 , R 4 and the C atom to which they are attached together form a cyclopropyl or cyclobutyl group;

[0113] Formula B can be obtained from doxorubicin by hydrolysis with dilute hydrochloric acid, protection of the protecting group, which can be prepared according to the method in the literature Jia M, et al. Synthesis of epirubicin hydrochloride. China Medical Industry Journal 47.5 (2016): 522-527.

[0114] Example 1: The synthesis method of Formula A is as follows.

[0115] Example 1.1 Synthesis of structure A1

[0116] (1) Synthesis of A1-1

[0117] Into a 250 mL single-necked flask, 2-amino-2-methyl-1-propanol (8.91 g, 100 mmol) and benzaldehyde (10.61 g, 100 mmol) were added, and then anhydrous ethanol (100 mL) was added. The reaction solution was stirred at 40 °C for 2 hours, then cooled to 0 °C in an ice bath, and sodium borohydride (4.92 g, 130 mmol) was added in portions. After 15 hours of reaction, the reaction was monitored by TLC, and then poured into a saturated ammonium chloride solution. Dichloromethane was used for extraction, and the organic phase was dried and purified by column chromatography (V / V, ethyl acetate / petroleum ether = 1 / 2) to obtain colorless transparent liquid A1-1 (5.74 g, yield 32.0%). LC-MS (ESI): [M+1] = 180.1.

[0118] (2) Synthesis of A1-2

[0119] Into a 250 mL single necked flask, A1-1 (5.74 g, 32.00 mmol) and toluene (50 mL) were added, then 2-bromo-isobutyric acid methyl ester (7.00 g, 38.40 mmol) was added, the reaction system was refluxed for 9 hours, TLC monitoring showed that the starting material was consumed, the reaction was stopped, the solution was concentrated under reduced pressure, the crude product was purified by column chromatography (V / V, ethyl acetate / petroleum ether = 1 / 3) to obtain white solid A1-2 (2.33 g, yield 29.5%).

[0120] (3) Synthesis of A1-3

[0121] A1-2 (2.33 g, 9.42 mmol) was dissolved in anhydrous methanol (50 mL) and stirred under ice water bath, then sodium triacetyl borohydride (5.99 g, 28.26 mmol) was added in batches, after stirring for 1 hour, the ice water bath was removed, and the reaction was carried out at room temperature for 10 hours, TLC monitoring showed that the starting material was consumed, the reaction solution was poured into saturated ammonium chloride solution, extracted with dichloromethane, the organic phase was dried and purified by column chromatography (V / V, ethyl acetate / petroleum ether = 1 / 2) to obtain colorless transparent liquid (1.08 g, yield 45.9%), the obtained oil was further separated by chiral separation to obtain colorless oil A1-3 (0.61 g, total yield of this step 25.9%).

[0122] LC-MS (ESI): [M+1]=250.3.

[0123] 1H NMR (400MHz, DMSO-d6) δ 7.20-7.51 (m, 5H), 5.82 (s, 1H), 5.21 (s, 1H), 3.72 (m, 2H), 3.62 (s, 2H), 1.21 (s, 12H).

[0124] The chiral separation conditions were as follows: Column: CHIRALPAK IG-3 (IG30CD-WE016), column size: 0.46 cm I.D. x 15 cm L, sample size: 1 ul, mobile phase: MeOH = 100%, flow rate: 1.0 ml / min, detection wavelength: UV 214 nm, column temperature: 35°C, HPLC equipment: Shimadzu LC-20AT CP-HPLC-06. The retention time of split peak 1 was 7.157 min; the retention time of split peak 2 was 8.152 min.

[0125] The synthesis of single chirality of A1-3 can be controlled by chiral reagents, the steps are as follows:

[0126] A 100 mL three-necked flask was dried thoroughly in an oven and then cooled to room temperature under nitrogen atmosphere. Under nitrogen protection, 10 mL of anhydrous THF and R-CBS solution (1 M THF solution) (0.5 mL, 0.05 eq) (CAS: 112022-83-0) were added to the reaction flask and cooled to -20 °C. BH3-THF (1.0 mL, 10.0 mmol) was diluted with 10 mL of anhydrous THF and then slowly added to the catalyst solution by syringe. After stirring for 10-15 minutes, the catalyst was allowed to pre-complex with borane. A1-2 (2.33 g, 9.42 mmol) was weighed into anhydrous THF (50 mL) and the above catalyst was transferred to a dropping funnel and slowly added to the reaction system, maintaining -20 °C for 1 hour. TLC was used to monitor the completion of the conversion of the starting material. The reaction flask was removed from the low-temperature bath. Methanol (MeOH) (10 mL) was slowly added dropwise with stirring. When no obvious bubbles were generated, stirring was continued for 20-30 minutes. To the mixture, 1 M aqueous HC1 (20 mL) was slowly added, and stirring was continued for 30 minutes. The reaction mixture was transferred to a separatory funnel and extracted with ethyl acetate (EtOAc) (30 mL x 3), and the combined organic phases were washed with saturated aqueous NaHC03(30 mL) and saturated brine (30 mL) in sequence. The combined organic phases were dried over anhydrous Na2S04and filtered to remove the drying agent. The solvent was removed by concentration under reduced pressure to obtain the crude product (light yellow oil), which was purified by column chromatography (V / V, ethyl acetate / petroleum ether = 1 / 2) to obtain colorless transparent liquid A1-3 (0.4 g, yield 16.9%). It was detected that the chiral synthesis product peak was consistent with peak 1.

[0127] LC-MS (ESI): [M+1] = 250.3.

[0128] 1H NMR (400 MHz, DMSO-d6) δ 7.20-7.51 (m, 5H), 5.82 (s, 1H), 5.21 (s, 1H), 3.72 (m, 2H), 3.62 (s, 2H), 1.21 (s, 12H).

[0129] (4) Synthesis of A1-4

[0130] A1-3 (0.61 g, 2.45 mmol) was dissolved in N,N-dimethylformamide (10 mL) under nitrogen atmosphere, sodium hydride (71.00 mg, 2.94 mmol) was added in batches under ice water bath condition, after 30 minutes of incubation and stirring, iodomethane (383.00 mg, 2.70 mmol) was added, the ice water bath was continued for 30 minutes of stirring, then the ice bath was removed and the reaction was continued at room temperature for 2 hours, the reaction was stopped after the raw material was completely converted by TLC monitoring, the reaction liquid was extracted with dichloromethane / water system, the organic phase was dried and concentrated to obtain a crude product, which was purified by column chromatography (V / V, ethyl acetate / petroleum ether = 1 / 3) to obtain white solid A1-4 (200.00 mg, yield 31.0%).

[0131] (5) Synthesis of A1-5

[0132] A1-4 (200.00 mg, 0.76 mmol) was dissolved in methanol (10 mL), nitrogen was replaced, palladium-carbon (5%, 7 mg) was added under nitrogen atmosphere, then the nitrogen in the system was replaced with a hydrogen balloon, after the replacement was completed, the system was stirred at room temperature for 6 hours, the raw material was completely converted by TLC monitoring, then it was filtered and the filtrate was concentrated to obtain a crude product, which was purified by column chromatography (V / V, ethyl acetate / petroleum ether = 1 / 2) to obtain colorless oil A1-5 (90.00 mg, yield 68.3%). LC-MS (ESI): [M+1]=174.2.

[0133] (6) Synthesis of A1

[0134] A1-5 (90.00 mg, 0.52 mmol) and A1-6 (CAS No.: 80483-18-7, commercially available, 97.00 mg, 0.57 mmol) were added to a 100 mL single-neck flask, then toluene (15 mL) was added, the reaction solution was stirred at 120°C for 2 hours, then it was cooled to room temperature, sodium borohydride (39.33 mg, 1.04 mmol) was added in batches, the reaction was continued at room temperature for 6 hours, the reaction was completed by TLC monitoring, the reaction liquid was concentrated to one third of the original volume, then it was poured into saturated ammonium chloride solution, extracted with dichloromethane, the organic phase was separated and dried, then it was purified by column chromatography (V / V, ethyl acetate / petroleum ether = 1 / 2) to obtain colorless oil A1 (80.64 mg, yield 49.5%). LC-MS (ESI): [M+1]=328.4.

[0135] Example 1.2 Synthesis of structure of formula A2

[0136] The synthesis of A2 was performed according to the procedure described for the synthesis of Al, replacing 2-amino-2-methyl-1-propanol with 1-aminocyclopropylmethanol (CAS No. 107017-72-1) and 2-bromoisobutyric acid methyl ester with 1-bromocyclopropane carboxylic acid methyl ester (CAS No. 96999-01-8). LC-MS (ESI): [M+1] = 310.0.

[0137] Example 1.3 Synthesis of structure A3

[0138] The synthesis of A3 was performed according to the procedure described for the synthesis of Al, replacing 2-amino-2-methyl-1-propanol with 1-aminocyclopropylmethanol. LC-MS (ESI): [M+1] = 312.2.

[0139] Example 1.4 Synthesis of structure A4

[0140] The synthesis of A4 was performed according to the procedure described for the synthesis of Al, replacing 2-bromoisobutyric acid methyl ester with 1-bromocyclopropane carboxylic acid methyl ester. LC-MS (ESI): [M+1] = 312.2.

[0141] Example 1.5 Synthesis of structure A5

[0142] The synthesis of A5 was performed according to the procedure described for the synthesis of Al, replacing iodomethane with iodoethane in the synthesis step.

[0143] LC-MS (ESI): [M+1] = 342.3.

[0144] Example 1.6 Synthesis of structure A6

[0145] The synthesis of A6 was performed according to the procedure described for the synthesis of Al, replacing iodomethane with bromocyclopropane and adding catalytic amount of sodium iodide for catalysis in the synthesis step. LC-MS (ESI): [M+1] = 354.6.

[0146] Example 1.7 Synthesis of structure A7

[0147] (1) Synthesis of A7-1

[0148] Under nitrogen atmosphere, hydroxyacetamide (0.75 g, 10 mmol) (CAS: 598-42-5) was dissolved in anhydrous THF, LiAlD4(0.50 g, 12 mmol) was added in batches under ice water bath, after 30 minutes of incubation and stirring, the ice bath was continued to stir for 30 minutes, the ice bath was removed, and the reaction was continued at room temperature for 2 hours, TLC monitoring reaction raw material conversion was completed, the reaction was stopped, the reaction liquid was extracted with ethyl acetate / water system, the organic phase was dried and concentrated to obtain the crude product, column chromatography purification (V / V, ethyl acetate / petroleum ether = 1 / 1) to obtain colorless oil A7-1 (0.52 g, yield 82.5%).

[0149] LC-MS (ESI): not shown

[0150] 1 H NMR (400 MHz, DMSO-d6) δ 8.76 (s, 2H), 3.82 (s, 2H)

[0151] (2) Synthesis of A7-2

[0152] In a 250 mL single-neck flask, A7-1 (0.52 g, 8 mmol) and benzaldehyde (1.07 g, 10 mmol) were added, and then anhydrous ethanol (10 mL) was added. The reaction solution was stirred at 40°C for 2 hours, then cooled to 0°C in an ice bath, and sodium borohydride (0.49 g, 13 mmol) was added in batches. After 15 hours of reaction, TLC monitoring showed that the reaction was complete. The reaction solution was poured into saturated ammonium chloride solution, extracted with dichloromethane, and the organic phase was separated and dried. Column chromatography purification (V / V, ethyl acetate / petroleum ether = 1 / 2) to obtain colorless transparent liquid A7-2 (1.0 g, yield 81.7%).

[0153] LC-MS (ESI): [M+1] = 154.3

[0154] 1 H NMR (400 MHz, DMSO-d6) δ 7.25-7.52 (m, 5H), 6.2 (s, 1H), 3.72 (s, 2H), 3.61 (s, 2H).

[0155] (3) Synthesis of A7-3

[0156] Into a 250 mL single necked flask, A7-2 (1.0 g, 6.5 mmol) and toluene (50 mL) were added, followed by methyl bromoacetate-D2 (3.0 g, 20 mmol) (CAS: 163886-16-6). The reaction was refluxed for 9 hours, and TLC was used to monitor the completion of the reaction. The solution was concentrated under reduced pressure, and the crude product was purified by column chromatography (V / V, ethyl acetate / petroleum ether = 1 / 3) to obtain white solid A7-3 (1.1 g, yield 86.7%).

[0157] (4) Synthesis of A7-4

[0158] A 100 mL three-necked flask was thoroughly dried in an oven and then cooled to room temperature under a nitrogen atmosphere. To the reaction flask, 10 mL of anhydrous THF and R-CBS solution (1 M THF solution) (0.5 mL, 0.1 eq) were added under nitrogen protection and cooled to -20 °C. BH3-THF (1.0 mL, 10.0 mmol) was diluted with 10 mL of anhydrous THF and then slowly added to the catalyst solution by syringe. After stirring for 10-15 minutes, the catalyst was allowed to pre-complex with borane. A7-3 (1.1 g, 6 mmol) was weighed and dissolved in anhydrous THF (50 mL), and the above catalyst was transferred to a dropping funnel and slowly added to the reaction system. After maintaining -20 °C for 1 hour, TLC was used to monitor the completion of the reaction. The reaction flask was removed from the low-temperature bath. With stirring, methanol (MeOH) (10 mL) was slowly added dropwise. When no obvious bubbles were generated, stirring was continued for 20-30 minutes. To the mixture, 1 M aqueous HCl (20 mL) was slowly added, and stirring was continued for 30 minutes. The reaction mixture was transferred to a separatory funnel and extracted with ethyl acetate (EtOAc) (3 x 30 mL), and the combined organic phases were washed with saturated aqueous NaHCO3 (30 mL) and saturated brine (30 mL), respectively. The combined organic phases were dried over anhydrous Na2SO4 and filtered to remove the drying agent. The solvent was removed by concentration under reduced pressure to obtain the crude product (pale yellow oil), which was purified by column chromatography (V / V, ethyl acetate / petroleum ether = 1 / 2) to obtain colorless transparent liquid A7-4 (0.9 g, yield 81%).

[0159] LC-MS (ESI): [M+1] = 198.4.

[0160] 1 H NMR (400 MHz, DMSO-d6) δ 7.20-7.51 (m, 5H), 5.82 (s, 1H), 5.21 (s, 1H), 3.72 (m, 2H), 3.60 (s, 2H).

[0161] (5) Synthesis of A7-5

[0162] A7-4 (0.9 g, 5 mmol) was dissolved in N,N-dimethylformamide (10 mL) under nitrogen atmosphere, sodium hydride (170.00 mg, 7 mmol) was added in batches under ice water bath, after 30 minutes of incubation and stirring, iodomethane (855 mg, 6 mmol) was added, the ice water bath was continued for 30 minutes, then the ice bath was removed and the reaction was continued at room temperature for 2 hours, TLC monitoring showed that the raw material was completely converted, the reaction was stopped, the reaction solution was extracted with dichloromethane / water system, the organic phase was dried and concentrated to obtain a crude product, which was purified by column chromatography (V / V, ethyl acetate / petroleum ether = 1 / 3) to obtain white solid A7-5 (0.8 g, yield 83.0%).

[0163] LC-MS (ESI): [M+1]= 212.5

[0164] (6) Synthesis of A7-6

[0165] A7-5 (0.8 g, 4 mmol) was dissolved in methanol (10 mL), replaced with nitrogen, and then palladium-carbon (5%, 40 mg) was added under nitrogen atmosphere, and then the nitrogen in the system was replaced with a hydrogen balloon. After the replacement was completed, the system was stirred at room temperature for 6 hours, TLC monitoring showed that the raw material was completely converted, then it was filtered and the filtrate was concentrated to obtain a crude product, which was purified by column chromatography (V / V, ethyl acetate / petroleum ether = 1 / 2) to obtain colorless oil A7-6 (0.4 g, yield 87.3%).

[0166] LC-MS (ESI): [M+1]= 122.3.

[0167] 1 H NMR (400 MHz, DMSO-d6) δ 5.5 (s, 1H), 3.72 (m, 2H), 3.43 (s, 3H)

[0168] (7) Synthesis of A7

[0169] A7-6 (0.4 g, 3.3 mmol) and A1-6 (1.7 g, 10 mmol) were added to a 100 mL single-neck flask, then toluene (15 mL) was added, the reaction solution was stirred at 120°C for 2 hours, then cooled to room temperature, sodium borohydride (950 mg, 25 mmol) was added in batches, and the reaction was continued at room temperature for 6 hours, TLC monitoring showed that the reaction was complete, the reaction solution was concentrated to one third of the original volume, poured into saturated ammonium chloride solution, extracted with dichloromethane, separated into organic and aqueous phases, the organic phase was dried, and then purified by column chromatography (V / V, ethyl acetate / petroleum ether = 1 / 2) to obtain colorless oil A7 (123 mg, yield 13.5%).

[0170] LC-MS (ESI): [M+1]= 276.3

[0171] 1 H NMR (400 MHz, DMSO-d6) δ 6.20-6.23 (m, 1H), 5.5 (s, 1H), 4.94 (m, 1H), 4.69 (m, 1H), 4.58 (m, 1H), 3.72 (m, 2H), 3.43 (s, 3H), 2.02 (s, 3H), 1.22 (m, 3H).

[0172] Synthesis of structure of Formula A8

[0173] Synthesis of Formula A8

[0174] (1) Synthesis of A8-1

[0175] Under nitrogen atmosphere, hydroxyacetaldehyde (0.6 g, 10 mmol) (CAS: 141-46-8) was dissolved in anhydrous EtOH (10 mL), BocNH2(1.4 g, 12 mmol) was added under ice water bath, then NaBD4(0.50 g, 12 mmol) was added in batches, after 30 minutes of incubation and stirring, the ice bath was continued to stir for 30 minutes, then the ice bath was removed, and the reaction was continued at room temperature for 2 hours, TLC monitoring reaction raw material conversion was completed, then 6M HC1 (3 mL) was added to the system, and stirred at room temperature for 2 hours. Stop the reaction, the reaction liquid was extracted with ethyl acetate / water system, the organic phase was dried and concentrated to obtain the crude product, which was purified by column chromatography (V / V, ethyl acetate / petroleum ether = 1 / 1) to obtain colorless oil A8-1 (0.62 g, yield 100%).

[0176] LC-MS (ESI): Not shown

[0177] 1 H NMR (400 MHz, DMSO-d6) δ 8.76 (s, 2H), 3.51-4.01 (m, 2H), 2.6 (t, J = 2 Hz, 1H)

[0178] (2) Synthesis of A8-2

[0179] In a 250 mL single-neck flask, A8-1 (0.62 g, 10 mmol) and benzaldehyde (1.27 g, 12 mmol) were added, and then anhydrous ethanol (10 mL) was added. The reaction solution was stirred at 40°C for 2 hours, then cooled to 0°C with ice bath, and sodium borohydride (0.57 g, 15 mmol) was added in batches. After 15 hours of reaction, TLC monitoring showed that the reaction was complete. Pour into saturated ammonium chloride solution, extract with dichloromethane, separate the phases, dry the organic phase, and purify by column chromatography (V / V, ethyl acetate / petroleum ether = 1 / 2) to obtain colorless transparent liquid A8-2 (1.5 g, yield 100%).

[0180] LC-MS (ESI): [M+1] = 153.1

[0181] 1 H NMR (400 MHz, DMSO-d6) δ 7.25 - 7.52 (m, 5H), 6.2 (s, 1H), 3.72 (s, 2H), 3.51 - 4.01 (m, 2H), 2.62 (t, J = 2 Hz, 1H).

[0182] (3) Synthesis of A8-3

[0183] Into a 250 mL single necked flask was placed A8-2 (1.5 g, 10 mmol) and toluene (50 mL), then 2-bromoethyl acetate-2-D (3.0 g, 20 mmol) (CAS: 1231247-95-2) was added, the reaction system was refluxed with water removal for 9 hours, TLC monitoring showed that the starting material was consumed, the reaction was stopped, the solution was concentrated under reduced pressure, the crude product was purified by column chromatography (V / V, ethyl acetate / petroleum ether = 1 / 3) to obtain white solid A8-3 (1.54 g, yield 79.8%).

[0184] LC-MS (ESI): [M+1] = 194.3

[0185] (4) Synthesis of A8-4

[0186] A 100 mL three-necked flask was dried thoroughly in an oven and then cooled to room temperature under a nitrogen atmosphere. To the reaction flask was added 10 mL of anhydrous THF and the R-CBS solution (1 M in THF) (0.5 mL, 0.1 eq) under nitrogen protection and cooled to -20 °C. BH3-THF (1.0 mL, 10.0 mmol) was diluted with 10 mL of anhydrous THF and added slowly to the catalyst solution via a syringe. The catalyst was allowed to pre-complex with borane by stirring for 10-15 min. A8-3 (1.54 g, 8 mmol) was weighed into anhydrous THF (50 mL) and the above catalyst was transferred to a dropping funnel and added slowly to the reaction mixture while maintaining the temperature at -20 °C for 1 h. TLC was used to monitor the completion of the reaction. The reaction flask was removed from the cold bath. Methanol (MeOH) (10 mL) was added slowly while stirring. When no more gas bubbles were produced, the stirring was continued for another 20-30 min. To the mixture was added slowly 1 M aqueous HC1 (20 mL) and stirred for 30 min. The reaction mixture was transferred to a separatory funnel and extracted with ethyl acetate (EtOAc) (3 x 30 mL) and the combined organic phase was washed with saturated aqueous NaHC03(30 mL) and saturated brine (30 mL) successively. The combined organic phase was dried over anhydrous Na2S04and filtered to remove the drying agent. The solvent was removed by concentration under reduced pressure to give the crude product (light yellow oil) which was purified by column chromatography (V / V, ethyl acetate / petroleum ether = 1 / 2) to give colorless transparent liquid A8-4 (1.21 g, 77.6% yield).

[0187] LC-MS (ESI): [M+1] = 196.3.

[0188] 1 H NMR (400 MHz, DMSO-d6) δ 7.20 - 7.51 (m, 5H), 5.82 (d, J = 7.0, 1H), 5.21 (s, 1H), 3.72 (m, 2H), 3.60 (s, 2H), 2.52 (m, 1H)

[0189] (5) Synthesis of A8-5

[0190] A8-4 (1.21 g, 6 mmol) was dissolved in N,N-dimethylformamide (10 mL) under nitrogen atmosphere, sodium hydride (190.00 mg, 8 mmol) was added in batches under ice water bath, after 30 minutes of incubation and stirring, iodomethane (1.0 g, 7 mmol) was added, and the stirring was continued for 30 minutes under ice water bath, then the ice bath was removed, and the reaction was continued at room temperature for 2 hours, the reaction was stopped after the raw material was completely converted by TLC monitoring, and the reaction solution was extracted with dichloromethane / water system, the organic phase was dried and concentrated to obtain a crude product, which was purified by column chromatography (V / V, ethyl acetate / petroleum ether = 1 / 3) to obtain white solid A8-5 (1.25 g, yield 96.4%).

[0191] LC-MS (ESI): [M+1] = 210.2

[0192] (6) Synthesis of A8-6

[0193] A8-5 (1.25 g, 6 mmol) was dissolved in methanol (10 mL), and nitrogen was replaced, then palladium-carbon (5%, 60 mg) was added under nitrogen atmosphere, and then the nitrogen in the system was replaced with a hydrogen balloon, after the replacement was completed, the stirring was continued at room temperature for 6 hours, the raw material was completely converted by TLC monitoring, and then the filtration was performed, and the filtrate was concentrated to obtain a crude product, which was purified by column chromatography (V / V, ethyl acetate / petroleum ether = 1 / 2) to obtain colorless oil A8-6 (0.71 g, yield 100%).

[0194] LC-MS (ESI): [M+1] = 120.3.

[0195] 1 H NMR (400 MHz, DMSO-d6) δ 5.82 (d, J = 7.0, 1H), 3.72 (m, 2H), 3.60 (s, 3H), 3.11 (m, 1H), 2.90 (m, 1H).

[0196] (7) Synthesis of A8

[0197] A8-6 (0.71 g, 6 mmol) and A1-6 (1.7 g, 10 mmol) were added to a 100 mL single-neck flask, and then toluene (15 mL) was added, the reaction solution was stirred at 120°C for 2 hours, and then cooled to room temperature, sodium borohydride (950 mg, 25 mmol) was added in batches, and the reaction was continued at room temperature for 6 hours, the reaction was completed by TLC monitoring, the reaction solution was concentrated to a volume of one third, poured into saturated ammonium chloride solution, extracted with dichloromethane, separated, the organic phase was dried, and purified by column chromatography (V / V, ethyl acetate / petroleum ether = 1 / 2) to obtain colorless oil A8 (350 mg, yield 21.3%).

[0198] LC-MS (ESI): [M+1] = 274.3

[0199] 1 H NMR (400 MHz, DMSO-d6) δ 6.20-6.23 (m, 1H), 5.5 (m, 1H), 4.94 (m, 1H), 4.69 (m, 1H), 4.58 (m, 1H), 3.72 (m, 2H), 3.62 (m, 1H), 3.43 (s, 3H), 3.11 (m, 1H), 2.90 (m, 1H), 2.02 (s, 3H), 1.22 (d, J = 2 Hz, 3H).

[0200] Example 1.9 Synthesis of structure of Formula A9

[0201] (1) Synthesis of A9-4

[0202] A 100 mL three-necked flask was thoroughly dried in an oven and then cooled to room temperature under nitrogen atmosphere. To the reaction flask was added 10 mL of anhydrous THF and S-CBS solution (1 M THF solution) (0.5 mL, 0.1 eq) (CAS: 112022-81-8) under nitrogen protection and cooled to -20 °C. BH3-THF (1.0 mL, 10.0 mmol) was diluted with 10 mL of anhydrous THF and then added slowly to the catalyst solution by syringe. After stirring for 10-15 minutes, the catalyst was allowed to pre-complex with borane. A7-3 (1.2 g, 6 mmol) was weighed and dissolved in anhydrous THF (50 mL), and the above catalyst was transferred to a dropping funnel and added slowly to the reaction system. After maintaining -20 °C for 1 hour, TLC was used to monitor the completion of the conversion of the starting material. The reaction flask was removed from the low-temperature bath. Methanol (MeOH) (10 mL) was added slowly with stirring. When no obvious gas bubbles were generated, stirring was continued for 20-30 minutes. To the mixture was slowly added 1 M aqueous HC1 (20 mL), and stirring was continued for 30 minutes. The reaction mixture was transferred to a separatory funnel and extracted with ethyl acetate (EtOAc) (3 x 30 mL), and the combined organic phases were washed with saturated aqueous NaHC03(30 mL) and saturated brine (30 mL) in sequence. The combined organic phases were dried over anhydrous Na2S04, and the drying agent was removed by filtration. The solvent was removed by concentration under reduced pressure to obtain the crude product (light yellow oil), which was purified using column chromatography (V / V, ethyl acetate / petroleum ether = 1 / 2) to obtain colorless transparent liquid A9-4 (481.1 mg, yield: 41.1%).

[0203] LC-MS (ESI): [M+1] = 198.4.

[0204] 1H NMR (400 MHz, DMSO-d6) δ 7.20-7.51 (m, 5H), 5.82 (s, 1H), 5.21 (s, 1H), 3.72 (m, 2H), 3.60 (s, 2H).

[0205] (2) Synthesis of A9-5

[0206] A9-4 (481.1 mg, 2.45 mmol) was dissolved in N,N-dimethylformamide (10 mL) under nitrogen atmosphere, sodium hydride (71.0 mg, 2.94 mmol) was added in batches under ice water bath condition, after 30 minutes of incubation and stirring, iodomethane (383.0 mg, 2.70 mmol) was added, after 30 minutes of continuous stirring under ice water bath, the ice bath was removed, and the reaction was continued at room temperature for 2 hours, TLC monitoring showed that the raw material was completely converted, the reaction was stopped, the reaction liquid was extracted with dichloromethane / water system, the organic phase was dried and concentrated to obtain a crude product, which was purified by column chromatography (V / V, ethyl acetate / petroleum ether = 1 / 3-1 / 2) to obtain colorless oil A9-5 (176.0 mg, yield 34.0%).

[0207] LC-MS (ESI): [M+1]=212.3.

[0208] (3) Synthesis of A9-6

[0209] A9-5 (176.0 mg, 0.83 mmol) was dissolved in methanol (10 mL) under nitrogen replacement, palladium on carbon (5%, 9.0 mg) was added under nitrogen atmosphere, then the nitrogen in the system was replaced with hydrogen balloon, after replacement, the mixture was stirred at room temperature for 6 hours, TLC monitoring showed that the raw material was completely converted, then it was filtered and the filtrate was concentrated to obtain a crude product, which was purified by column chromatography (V / V, ethyl acetate / petroleum ether = 1 / 1-2 / 1) to obtain colorless oil A9-6 (62.1 mg, yield 61.8%).

[0210] LC-MS (ESI): [M+1]=122.2.

[0211] (6) Synthesis of A9

[0212] A9-6 (62.1 mg, 0.51 mmol) and A1-6 (CAS No.: 80483-18-7, commercially available, 97.0 mg, 0.57 mmol) were added into a 100 mL single neck flask, then toluene (15 mL) was added, the reaction solution was stirred at 120 °C for 2 hours, then cooled to room temperature, sodium borohydride (39.3 mg, 1.04 mmol) was added in batches, the reaction was continued at room temperature for 6 hours, TLC monitoring showed that the reaction was completed, the reaction solution was concentrated to one third of the original volume, poured into saturated ammonium chloride solution, extracted with dichloromethane, separated phases, dried the organic phase, purified by column chromatography (V / V, ethyl acetate / petroleum ether = 1 / 2) to obtain colorless oil A9 (45.2 mg, yield 32.0%).

[0213] LC-MS (ESI): [M+1] = 276.3.

[0214] 1 H NMR (400 MHz, DMSO-d6) δ 6.20-6.23 (m, 1H), 5.5 (s, 1H), 4.94 (m, 1H), 4.69 (m, 1H), 4.56 (m, 1H), 3.69 (m, 2H), 3.43 (s, 3H), 2.02 (s, 3H), 1.22 (m, 3H).

[0215] Example 1.10 Synthesis of structure of formula A10

[0216] (1) Synthesis of A10-4

[0217] A 100 mL three-necked flask was thoroughly dried in an oven and then cooled to room temperature under a nitrogen atmosphere. To the reaction flask was added 10 mL of anhydrous THF and the S-CBS solution (1 M in THF) (0.5 mL, 0.1 eq) under nitrogen protection and cooled to -20 °C. BH3-THF (1.0 mL, 10.0 mmol) was diluted with 10 mL of anhydrous THF and added slowly to the catalyst solution via a syringe. The catalyst was allowed to pre-complex with borane by stirring for 10-15 min. A8-3 (1.54 g, 8 mmol) was weighed into anhydrous THF (50 mL) and the above catalyst was transferred to a dropping funnel and added slowly to the reaction mixture, maintaining the temperature at -20 °C for 1 h. TLC was used to monitor the completion of the reaction. The reaction flask was removed from the cold bath. Methanol (MeOH) (10 mL) was added slowly with stirring. When no more gas bubbles were produced, the stirring was continued for another 20-30 min. To the mixture was added slowly 1 M aqueous HC1 (20 mL) and stirred for 30 min. The reaction mixture was transferred to a separatory funnel and extracted with ethyl acetate (EtOAc) (3 x 30 mL). The combined organic phase was washed with saturated aqueous NaHC03(30 mL) and saturated brine (30 mL) successively. The combined organic phase was dried over anhydrous Na2S04and filtered to remove the drying agent. The solvent was removed by concentration under reduced pressure to give the crude product (light yellow oil). Column chromatography (V / V, ethyl acetate / petroleum ether = 1 / 2) was used to purify the product to give colorless transparent liquid A10-4 (1.51 g, 97% yield).

[0218] LC-MS (ESI): [M+1] = 196.3.

[0219] 1 H NMR (400 MHz, DMSO-d6) δ 7.20 - 7.51 (m, 5H), 5.82 (d, J = 7.0, 1H), 5.21 (s, 1H), 3.72 (m, 2H), 3.60 (s, 2H), 2.52 (m, 1H)

[0220] (2) Synthesis of A10-5

[0221] A10-4 (1.51 g, 8 mmol) was dissolved in N,N-dimethylformamide (10 mL) under nitrogen atmosphere, sodium hydride (243 mg, 10 mmol) was added in batches under ice water bath, after 30 minutes of incubation and stirring, iodomethane (1.4 g, 10 mmol) was added, and the stirring was continued for 30 minutes under ice water bath, then the ice bath was removed, and the reaction was continued for 2 hours at room temperature, the reaction was stopped after the raw material was completely converted by TLC monitoring, and the reaction solution was extracted with dichloromethane / water system, the organic phase was dried and concentrated to obtain a crude product, which was purified by column chromatography (V / V, ethyl acetate / petroleum ether = 1 / 3) to obtain white solid A10-5 (2.0 g, yield 96%).

[0222] LC-MS (ESI): [M+1] = 210.2

[0223] (3) Synthesis of A10-6

[0224] A10-5 (2.0 g, 10 mmol) was dissolved in methanol (10 mL), and nitrogen was replaced, then palladium-carbon (5%, 100 mg) was added under nitrogen atmosphere, and then the nitrogen in the system was replaced with a hydrogen balloon, after the replacement was completed, the stirring was continued for 6 hours at room temperature, the raw material was completely converted by TLC monitoring, and then the filtration was performed, and the filtrate was concentrated to obtain a crude product, which was purified by column chromatography (V / V, ethyl acetate / petroleum ether = 1 / 2) to obtain colorless oil A10-6 (1.2 g, yield 100%).

[0225] LC-MS (ESI): [M+1] = 120.3.

[0226] 1 H NMR (400 MHz, DMSO-d6) δ 5.82 (d, J = 7.0, 1H), 3.72 (m, 2H), 3.60 (s, 3H), 3.11 (m, 1H), 2.90 (m, 1H).

[0227] (4) Synthesis of A10

[0228] A10-6 (1.2 g, 10 mmol) and A1-6 (2.1 g, 12 mmol) were added to a 100 mL single-neck flask, and then toluene (100 mL) was added, the reaction solution was stirred at 120°C for 2 hours, and then cooled to room temperature, sodium borohydride (950 mg, 25 mmol) was added in batches, and the reaction was continued at room temperature for 6 hours, the reaction was completed by TLC monitoring, the reaction solution was concentrated to a volume of one third, poured into saturated ammonium chloride solution, extracted with dichloromethane, separated, the organic phase was dried, and purified by column chromatography (V / V, ethyl acetate / petroleum ether = 1 / 2) to obtain colorless oil A10 (653 mg, yield 23.9%).

[0229] LC-MS (ESI): [M+1] = 274.5

[0230] 1 H NMR (400 MHz, DMSO-d6) δ 6.20-6.23 (m, 1H), 5.5 (m, 1H), 4.94 (m, 1H), 4.69 (m, 1H), 4.58 (m, 1H), 3.72 (m, 2H), 3.62 (m, 1H), 3.43 (s, 3H), 3.11 (m, 1H), 2.90 (m, 1H), 2.02 (s, 3H), 1.22 (d, J = 2 Hz, 3H).

[0231] Example 2: The compound of the application (Formula D) was synthesized as follows:

[0232] Example 2.1 Synthesis of structure D1

[0233] B (326.00 mg, 0.50 mmol), A1 (171.00 mg, 0.60 mmol) and p-toluenesulfonic acid (172.20 mg, 1.00 mmol) were weighed into a 100 mL single-necked flask, 10 mL of toluene was added, the system was warmed to 50°C for 5 hours, TLC monitoring showed that the starting material was completely converted, and the crude product was obtained by reducing pressure concentration;

[0234] The crude product from the previous step was dissolved in 15 mL of tetrahydrofuran, 1N tetrabutylammonium fluoride solution in tetrahydrofuran (3.00 mL, 3.00 mmol) was added, and the system was stirred at room temperature for 5 hours. Then 2N potassium carbonate solution (1 mL) was added dropwise, and the reaction was continued for 3 hours. TLC monitoring showed that the starting material was completely converted, and the crude product was obtained by reducing pressure concentration to about one-third of the original volume. The product was extracted with ethyl acetate / water, and the organic phase was dried and concentrated to obtain the crude product. The crude product was purified by column chromatography (V / V, dichloromethane / methanol = 20 / 1) to obtain brown-red solid D1 (115 mg, two-step yield 33.0%).

[0235] LC-MS (ESI): [M+1] = 700.4

[0236] 1H NMR (400 MHz, DMSO-d6) δ 14.00 (s, 1H), 13.21 (s, 1H), 7.90 - 7.82 (m, 2H), 7.63 (dd, J = 7.6, 2.1 Hz, 1H), 5.53 - 5.40 (m, 2H), 5.29 (d, J = 2.7 Hz, 1H), 5.27 (s, 1H), 4.96 - 4.79 (m, 2H), 4.59 (d, J = 5.8 Hz, 2H), 4.18 (t, J = 6.7 Hz, 1H), 3.98 (s, 3H), 3.67 - 3.42 (m, 2H), 3.61 (d, J = 4.6 Hz, 1H), 3.40 (s, 3H), 3.36 (d, J = 13.1 Hz, 1H), 3.03 - 2.81 (m, 2H), 2.23 - 2.04 (m, 2H), 1.89 (td, J = 12.6, 3.5 Hz, 1H), 1.69 (dd, J = 12.1, 4.1 Hz, 1H), 1.20 (s, 12H), 1.16 (d, J = 6.5 Hz, 3H).

[0237] Example 2.2 Synthesis of structure D3

[0238] The synthesis was performed according to Example 2.1 with the substitution of A1 by A2 to give D3 as a brownish red solid (64.35 mg, 18.5% yield over two steps).

[0239] LC-MS (ESI): [M+1] = 696.8

[0240] 1 H NMR (400 MHz, DMSO-d6) δ 14.00 (s, 1H), 13.21 (s, 1H), 7.90 - 7.82 (m, 2H), 7.63 (dd, J = 7.6, 2.1 Hz, 1H), 5.53 - 5.40 (m, 2H), 5.29 (d, J = 2.7 Hz, 1H), 5.27 (s, 1H), 4.96 - 4.79 (m, 2H), 4.59 (d, J = 5.8 Hz, 2H), 4.18 (t, J = 6.7 Hz, 1H), 3.98 (s, 3H), 3.67 - 3.42 (m, 2H), 3.61 (d, J = 4.6 Hz, 1H), 3.40 (s, 3H), 3.36 (d, J = 13.1 Hz, 1H), 3.03 - 2.81 (m, 2H), 2.23 - 2.04 (m, 2H), 1.89 (td, J = 12.6, 3.5 Hz, 1H), 1.69 (dd, J = 12.1, 4.1 Hz, 1H), 1.16 (d, J = 6.5 Hz, 3H), 0.46 - 0.21 (m, 8H).

[0241] Example 2.3 Synthesis of structure D5

[0242] The synthesis was performed according to the procedure described in Example 2.1, replacing Al with A4, to give D5 as a brownish red solid (58.61 mg, 16.8% yield over two steps).

[0243] LC-MS (ESI): [M+1] = 698.9

[0244] 1 H NMR (400 MHz, DMSO-d6) δ 14.03 (s, 1H), 13.19 (s, 1H), 7.90 - 7.85 (m, 2H), 7.65 (dd, J = 7.6, 2.1 Hz, 1H), 5.55 - 5.40 (m, 2H), 5.29 (d, J = 2.7 Hz, 1H), 5.27 (s, 1H), 4.96 - 4.79 (m, 2H), 4.59 (d, J = 5.8 Hz, 2H), 4.18 (t, J = 6.7 Hz, 1H), 3.98 (s, 3H), 3.67 - 3.42 (m, 2H), 3.60 (d, J = 4.6 Hz, 1H), 3.40 (s, 3H), 3.36 (d, J = 13.1 Hz, 1H), 3.03 - 2.81 (m, 2H), 2.23 - 2.04 (m, 2H), 1.89 (td, J = 12.6, 3.5 Hz, 1H), 1.69 (dd, J = 12.1, 4.1 Hz, 1H), 1.16 (d, J = 6.5 Hz, 3H), 1.10 (s, 6H), 0.46 - 0.21 (m, 4H).

[0245] Example 2.4 Synthesis of structure D6

[0246] The synthesis was performed according to the procedure described in Example 2.1, replacing Al with A3, to give D6 as a brownish red solid (143.03 mg, 20.5% yield over two steps).

[0247] LC-MS (ESI): [M+1] = 698.9

[0248] 1H NMR (400 MHz, DMSO-d6) δ 14.05 (s, 1H), 13.20 (s, 1H), 7.93 - 7.80 (m, 2H), 7.63 (dd, J = 7.6, 2.1 Hz, 1H), 5.50 - 5.40 (m, 2H), 5.27 (d, J = 2.7 Hz, 1H), 5.25 (s, 1H), 4.97 - 4.80 (m, 2H), 4.58 (d, J = 5.8 Hz, 2H), 4.18 (t, J = 6.7 Hz, 1H), 3.98 (s, 3H), 3.67-3.42 (m, 2H), 3.61 (d, J = 4.6 Hz, 1H), 3.40 (s, 3H), 3.36 (d, J = 13.1 Hz, 1H), 3.03 - 2.81 (m, 2H), 2.25 - 2.05 (m, 2H), 1.87 (td, J = 12.6, 3.5 Hz, 1H), 1.69 (dd, J = 12.1, 4.1 Hz, 1H), 1.16 (d, J = 6.5 Hz, 3H), 1.13 (s, 6H) 0.48 - 0.29 (m, 4H).

[0249] Example 2.5. Synthesis of structure D7 of Formula D

[0250] The synthesis method was referenced to Example 2.1, replacing A1 with A5, to give D7 as a brownish red solid (75.00 mg, 21.0% yield over two steps).

[0251] LC-MS (ESI): [M+1] = 714.9

[0252] 1H NMR (400 MHz, DMSO-d6) δ 14.00 (s, 1H), 13.21 (s, 1H), 7.90 - 7.82 (m, 2H), 7.63 (dd, J = 7.8, 2.1 Hz, 1H), 5.53 - 5.40 (m, 2H), 5.29 (d, J = 2.8 Hz, 1H), 5.27 (s, 1H), 4.96 - 4.79 (m, 2H), 4.59 (d, J = 5.9 Hz, 2H), 4.18 (t, J = 6.7 Hz, 1H), 3.88 (q, J = 7.5 Hz, 2H), 3.67 - 3.42 (m, 2H), 3.61 (d, J = 4.6 Hz, 1H), 3.40 (s, 3H), 3.36 (d, J = 13.1 Hz, 1H), 3.03 - 2.81 (m, 2H), 2.23 - 2.04 (m, 2H), 1.89 (td, J = 12.6, 3.5 Hz, 1H), 1.69 (dd, J = 12.1, 4.1 Hz, 1H), 1.23 (t, J = 7.5 Hz, 3H), 1.20 (s, 12H), 1.16 (d, J = 6.4 Hz, 3H).

[0253] Example 2.6. Synthesis of structure D8

[0254] The synthesis was performed according to the procedure described in Example 2.1, replacing A1 with A6, to give D8 as a brownish red solid (60.24 mg, 16.6% yield over two steps).

[0255] LC-MS (ESI): [M+1] = 726.8

[0256] 1H NMR (400 MHz, DMSO-d6) δ 14.02 (s, 1H), 13.18 (s, 1H), 7.88 - 7.82 (m, 2H), 7.63 (dd, J = 7.8, 2.1 Hz, 1H), 5.53 - 5.40 (m, 2H), 5.29 (d, J = 2.8 Hz, 1H), 5.27 (s, 1H), 4.96 - 4.79 (m, 2H), 4.59 (d, J = 5.9 Hz, 2H), 4.18 (t, J = 6.7 Hz, 1H), 3.37 (m, 1H), 3.67 - 3.42 (m, 2H), 3.61 (d, J = 4.6 Hz, 1H), 3.40 (s, 3H), 3.36 (d, J = 13.1 Hz, 1H), 3.03 - 2.81 (m, 2H), 2.23 - 2.04 (m, 2H), 1.89 (td, J = 12.6, 3.5 Hz, 1H), 1.69 (dd, J = 12.1, 4.1 Hz, 1H), 1.20 (s, 12H), 1.16 (d, J = 6.4 Hz, 3H), 0.58 - 0.34 (m, 4H).

[0257] Example 2.7. Synthesis of structure D9

[0258] The synthesis was performed according to the procedure described in Example 2.1 by replacing A1 with A7 to give D9 as a brownish red solid.

[0259] LC-MS (ESI): [M+1] = 648.4,

[0260] 1H NMR (400 MHz, DMSO-d6) δ 13.96 (s, 1H), 13.16 (s, 1H), 7.96 - 7.73 (m, 2H), 7.58 (d, J = 9.3 Hz, 1H), 5.29 (d, J = 18.8 Hz, 2H), 4.96 - 4.87 (m, 1H), 4.83 (t, J = 5.9 Hz, 1H), 4.56 (d, J = 6.1 Hz, 2H), 4.35 (d, J = 9.1 Hz, 1H), 4.09 (d, J = 5.2 Hz, 2H), 3.95 (s, 3H), 3.73 (dd, J = 11.1, 4.5 Hz, 1H), 3.59 (s, 1H), 3.41 (d, J = 11.3 Hz, 1H), 3.26 (s, 3H), 2.90 (s, 2H), 2.43 (d, J = 10.1 Hz, 1H), 2.18 (dd, J = 14.3, 2.8 Hz, 1H), 2.10 (dd, J = 14.3, 5.5 Hz, 1H), 1.82 (t, J = 14.4 Hz, 1H), 1.59 (d, J = 9.6 Hz, 1H), 1.14 (d, J = 6.5 Hz, 3H).

[0261] Example 2.8 Synthesis of structure D11

[0262] The synthesis method refers to Example 2.1, replacing A1 with A8 to obtain red solid D11.

[0263] LC-MS (ESI): [M+1] = 646.3,

[0264] 1 H NMR (CDCI3, 400 MHz): δ (ppm) 13.99 (s, 1H), 13.26 (s, 1H), 8.05 (d, J = 7.6 Hz, 1H), 7.79 (t, J = 8.1 Hz, 1H), 7.41 (d, J = 8.5 Hz, 1H), 5.57 (s, 1H), 5.31 (s, 1H), 4.74 (d, J = 13.0 Hz, 3H), 4.51 (s, 1H), 4.09 (s, 3H), 3.96 (d, J = 6.7 Hz, 2H), 3.701 (s, 1H), 3.57 (s, 1H), 3.39 (s, 3H), 3.30 (d, J = 18.4 Hz, 1H), 3.07 - 3.00 (m, 3H), 2.58 - 2.36 (m, 4H), 2.17 (dd, J = 14.6, 3.6 Hz, 1H), 1.79 (s, 2H), 1.36 (d, J = 6.5 Hz, 3H).

[0265] Example 2.9 Synthesis of structure D12

[0266] The synthesis was performed according to reference example 2.1, replacing A1 by A9, to give D12 as a brownish red solid.

[0267] LC-MS (ESI): [M+1] = 648.4,

[0268] 1 H NMR (400 MHz, DMSO-d6) δ 13.96 (s, 1H), 13.16 (s, 1H), 7.96 - 7.73 (m, 2H), 7.58 (d, J = 9.3 Hz, 1H), 5.29 (d, J = 18.8 Hz, 2H), 4.96 - 4.87 (m, 1H), 4.83 (t, J = 5.9 Hz, 1H), 4.56 (d, J = 6.1 Hz, 2H), 4.35 (d, J = 9.1 Hz, 1H), 4.09 (d, J = 5.2 Hz, 2H), 3.95 (s, 3H), 3.73 (dd, J = 11.1, 4.5 Hz, 1H), 3.59 (s, 1H), 3.41 (d, J = 11.3 Hz, 1H), 3.26 (s, 3H), 2.90 (s, 2H), 2.43 (d, J = 10.1 Hz, 1H), 2.18 (dd, J = 14.3, 2.8 Hz, 1H), 2.10 (dd, J = 14.3, 5.5 Hz, 1H), 1.82 (t, J = 14.4 Hz, 1H), 1.59 (d, J = 9.6 Hz, 1H), 1.14 (d, J = 6.5 Hz, 3H).

[0269] Synthesis of the structure of formula D13 Example 2.10

[0270] The synthesis was performed according to reference example 2.1, replacing A1 by A10, to give D13 as a brown solid.

[0271] LC-MS (ESI): [M+1] = 646.3, 1H NMR (CDC13, 400 MHz): δ (ppm) 13.99 (s, 1H), 13.26 (s, 1H), 8.05 (d, J = 7.6 Hz, 1H), 7.79 (t, J = 8.1 Hz, 1H), 7.41 (d, J = 8.5 Hz, 1H), 5.57 (s, 1H), 5.31 (s, 1H), 4.74 (d, J = 13.0 Hz, 3H), 4.51 (s, 1H), 4.09 (s, 3H), 3.96 (d, J = 6.7 Hz, 2H), 3.701 (s, 1H), 3.57 (s, 1H), 3.39 (s, 3H), 3.30 (d, J = 18.4 Hz, 1H), 3.07-3.00 (m, 3H), 2.58-2.36 (m, 4H), 2.17 (dd, J = 14.6, 3.6 Hz, 1H), 1.79 (s, 2H), 1.36 (d, J = 6.5 Hz, 3H).

[0272] Inhibitory activity of Example 3 on different tumor cells

[0273] (1) The effect of anthracycline compounds on the proliferation of tumor cells in vitro was detected by SRB method for adherent cells. A certain number of cells in logarithmic growth phase were inoculated in 96-well culture plates, and adherent growth was performed overnight, then different concentrations of anthracycline compounds were added. After 144 hours, fixation was performed with trichloroacetic acid. After staining with SRB (prepared with 1% glacial acetic acid, concentration 4 mg / mL), 10 mM Tris solution was added to each well for dissolution, and the OD value was read on an enzyme marker at a wavelength of 510 nm.

[0274] Inhibition rate (%) = (OD value of control well - OD value of drug administration well) / OD value of control well x 100%

[0275] (2) The effect of anthracycline compounds on the proliferation of tumor cells in vitro was detected by MTT method for suspension cells. A certain number of cells in logarithmic growth phase were inoculated in 96-well culture plates, and adherent growth was performed overnight, then different concentrations of anthracycline compounds were added. After 144 hours, MTT was added to each well, and incubation was continued in a 5% CO2 saturated humidity incubator at 37°C for 4 hours. 10 μL of triple liquid (sodium dodecyl sulfate 10 g, isobutyl alcohol 5 ml, 10 mol / L hydrochloric acid solution 0.1 mL, dissolved with double distilled water to make 100 mL solution) was added to each well, and the OD value was determined on an enzyme marker at a wavelength of 570 nm and 690 nm.

[0276] (3) The half inhibitory concentration IC50 was calculated with Graphpad Prism 8.0 software according to the inhibition rate of each concentration obtained from the above experiment. 50 The experiment was repeated 1 time independently, and the data was expressed as mean SD, and the results were shown in Table 1.

[0277] Table 1 Inhibitory activity of anthracycline series compounds on different tumor cells

[0278] (wherein * indicates the same batch of tests)

[0279] The present application analyzes the relevant tumor inhibitory activity of the patent compounds by different tumor activity inhibition experiments. The compounds (D1-D10) of the present application can maintain a good level of relevant tumor inhibitory activity comparable to that of nemonabix, and are higher than that of doxorubicin and daunorubicin and other anthracycline compounds.

[0280] Example 4 Metabolic substance analysis of anthracycline series compounds in rat liver microsomes

[0281] (1) Preparation of rat liver microsomes

[0282] Sprague-Dawley rats, male, body weight 250-280 grams, after 12 hours of the last dose, the animals were fasted, free water, after 12 hours of fasting, the animals were decapitated, laparotomy, the liver was taken out, washed with ice physiological saline, filter paper was used to absorb the water, the liver was cut into small pieces with scissors, washed with ice-cold sucrose solution for 2-3 times until the washing liquid was colorless or light yellow. Cut with scissors, then add about 4 times the liver weight of sucrose solution, homogenate in ice bath for 20 s, repeat 2 times, transfer the homogenate to centrifuge tube, high speed centrifuge at 16000 g for 20 minutes (4℃), take the supernatant to ultracentrifuge at 100000 g for 60 minutes (4℃), discard the supernatant, the precipitate was suspended with potassium pyrophosphate solution, ultracentrifuged again at 100000 g for 60 minutes (4℃), discard the supernatant, the precipitate was suspended with 2 times the liver weight of Tris-HCL buffer, aliquot, stored at 80℃ for standby.

[0283] (2) Incubation system of rat liver microsomes and sample treatment

[0284] Take the above prepared liver microsomes of appropriate volume, add phosphate buffer (pH = 7.4) to prepare a solution with a final concentration of 1 mg / mL, take 5 mL; then prepare a DMF solution (1 mg / mL) of the target compound, dissolve in the liver microsome solution, mix and shake well, then pre-warm in a 37℃ water bath for 3 hours, after incubation, dry, dissolve the residue in 200 μL of methanol for testing.

[0285] (3) Analysis conditions

[0286] Analysis column: Hypersil BDS C18 column (2.1 mm X 100 mm, 3 μm), mobile phase: methanol: water containing 1% formic acid = 9:1, determined by electrospray ion source (ESI) positive ion mode under the condition of first full scan, second mass spectrometry in two ways at the same time. Mass spectrometry conditions: spray voltage 3500V, sheath gas pressure 35psi, auxiliary gas pressure 11psi, capillary temperature 350°C.

[0287] Table 2: Description of samples to be tested

[0288] The results of the metabolite analysis of anthracycline series compounds in rat liver microsomes are shown in Table 3.

[0289] Table 3: Metabolite analysis of anthracycline series compounds in rat liver microsomes

[0290] The present application analyzes the metabolites produced by the metabolism of the patent compound in the liver through in vitro simulation experiments of rat liver microsomes, and analyzes the metabolites by mass spectrometry data. No metabolites similar to the structure of PNU-159682 are produced.

[0291] Example 4: Pharmacokinetic test of candidate compound

[0292] Description of test samples

[0293] Preparation of test solution:

[0294] Dissolve the test sample (TA) into a stock solution. Take an appropriate amount of TA and dissolve it in 5% DMSO + 95% (20% HP-β-CD) to obtain the required solution for oral administration (PO).

[0295] For oral administration to mice, fasted male mice (n=3 per group) received 50 mg / kg of test substance at a dose of 10 mL / kg by oral gavage. Plasma was collected in K2-EDTA tubes from 0.0083 hours to 24 hours (0.083, 0.25, 0.5, 1, 2, 4, 8, and 24 hours after administration) by the saphenous vein (continuous sampling). For oral administration to rats, fasted male SD rats (n=3 per group per time point) received 50 mg / kg of IPG7236 at a dose of 10 mL / kg by oral gavage, with 5% DMSO + 95% (20% HP-β-CD) as the solvent. Blood was collected into K2-EDTA tubes by the jugular vein and processed into plasma.

[0296] The analyte levels in plasma were analyzed by liquid chromatography-tandem mass spectrometry (LC-MS / MS) (API-4000 (LC-MS / MS) system, equipped with Analyst software 1.6.3, Chromatographic column: ACQUITY UPLC BEH C18 2.1 x 50, 1.7 μ). Briefly, 20 μL of plasma sample was taken and protein precipitated with 400 μL of methanol containing 100 ng / mL of internal standard. The mixture was vortexed for 1 minute and then centrifuged at 18000 g for 7 minutes. 200 μL of supernatant was transferred to a 96-well plate. 1 μL of supernatant was taken for LC-MS / MS analysis.

[0297] Blood sample collection and procedure

[0298] Blood was collected by jugular vein or other suitable vein, 0.2 mL each time. The sample was placed in a tube containing K2-EDTA and stored on ice before centrifugation. Within 1 hour after collection, the blood sample was centrifuged at 6800 g for 6 minutes, with temperature control at 2-8°C, and the sample was stored frozen at about -80°C.

[0299] Sample analysis and data processing

[0300] The analytical results were confirmed using quality control samples to assess intra-batch variability. The accuracy of more than 66.7% of the quality control samples was between 80-120% of the known value.

[0301] The study director calculated a series of standard parameters, including area under the curve (AUC(0-t) and AUC(0-∞)), elimination half-life (T1 / 2), maximum plasma concentration (Cmax), and time to maximum plasma concentration (Tmax), using the non-compartmental analysis module in the U.S. Food and Drug Administration (FDA)-certified pharmacokinetic program Phoenix WinNonlin 7.0 (Pharsight, USA).

[0302] The final test results are as follows

[0303] Table 4 Pharmacokinetic data of the test compound

[0304] From the pharmacokinetic results, through structural optimization, we obtained D1, D9 and D11, which, compared with the prototype drug nalmufenibacin, increased the in vivo exposure by 2-10 times and the oral bioavailability by 5-7 times at the same dose.

[0305] Compared with the prior art, the derivative of nemonoxit has the following beneficial effects: effectively prevents rapid metabolism of the drug in the body, especially inhibits the conversion path to the high-activity metabolite PNU-159682 structure; significantly improves the metabolic stability and oral bioavailability, and the compound of the present application maintains good tumor inhibition activity. That is, compared with nemonoxit, the compound of the present application not only realizes the improvement of metabolic stability and oral bioavailability, but also maintains excellent tumor inhibition activity, ensuring the treatment effect.

Claims

1. An anthracenedione compound of Formula I: ###0001### or a pharmaceutically acceptable salt thereof. R 1 and R 2 are independently H, F, deuterium, C 1-6 alkyl, or R 1 , R 2 and the C atom to which they are attached together form a cyclopropyl, cyclobutyl, cyclopentyl, or cyclohexyl; and R 1 and R 2 are not simultaneously H; R 3 and R 4 are independently H, F, deuterium, C 1-6 alkyl, or R 3 , R 4 and the C atom to which they are attached together form a cyclopropyl, cyclobutyl, cyclopentyl, or cyclohexyl; and R 3 and R 4 are not both H; R 5 is hydrogen, C 1-6 alkyl, cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl.

2. The anthracycline compound of formula I as claimed in claim 1, characterized in that, which satisfies one or more of the following conditions: (1) R 1 (2) R 2 (3) R 3 (4) R 4 (5) R 5 (6) R 1-6 alkyl is independently methyl, ethyl, n-propyl, i-propyl, n-butyl, s-butyl, i-butyl, or t-butyl; for example, methyl or ethyl; for example, methyl. (2) R 1 and R 2 , R 1 , R 2 and the C atom to which they are attached together form a cyclopropyl or cyclobutyl group, for example cyclopropyl; and (3) R 3 and R 4 , R 3 , R 4 and the C atom to which they are attached together form a cyclopropyl or cyclobutyl group, for example cyclopropyl.

3. The anthracycline compound of claim 1, or a pharmaceutically acceptable salt thereof, having the formula: ###0002### wherein R is H, OH, or OCH3. which satisfies one or more of the following conditions: (1) R 1 and R 2 are independently C 1-6 alkyl, or R 1 , R 2 and the C atom to which they are attached together form a cyclopropyl or cyclobutyl group; preferably R 1 and R 2 are independently methyl, or R 1 , R 2 and the C atom to which they are attached together form a cyclopropyl group; (2) R 3 and R 4 independently are C 1-6 alkyl, or R 3 , R 4 and the C atom to which they are attached together form a cyclopropyl or cyclobutyl group; preferably, R 3 and R 4 independently are methyl, or R 3 , R 4 and the C atom to which they are attached together form a cyclopropyl group; and (3) R 5 is C 1-6 alkyl, cyclopropyl or cyclobutyl; preferably, R 5 is C 1-6 alkyl, for example methyl.

4. The anthracycline compound of claim 1, or a pharmaceutically acceptable salt thereof, having the formula: ###0001### wherein R is H, OH, or OCH3. which satisfies one or more of the following conditions: (1) R 1 is C 1-6 alkyl or R 1 is H or deuterium; (2) R 2 is F, deuterium or C 1-6 alkyl, preferably R 2 is C 1-6 alkyl or R 2 is deuterium; (3) R 3 is C 1-6 alkyl or R 3 is H or deuterium; (4) R 4 is F, deuterium or C 1-6 alkyl, preferably R 4 is C 1-6 alkyl or R 4 is deuterium; (5) R 1 and R 2 are the same; (6) R 3 and R 4 are the same; and (7) the anthracycline compound as shown in Formula I is an anthracycline compound as shown in Formula I-X or I-Y, R 5 is C 1-6 alkyl, cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl; Preferably, in the anthracene ring compound of formula I, R 1 , R 2 , R 3 and R 4 are the same.

5. The anthracycline compound of claim 1, or a pharmaceutically acceptable salt thereof, having the formula: ###0002### wherein R is H, OH, or OCH3. which satisfies one or more of the following conditions: (1) R 1 is H, ethyl, F, or deuterium; or, R 1 is H, methyl, or deuterium; (2) R 2 is ethyl, F or deuterium; or, R 2 is methyl or deuterium; (3) R 3 is H, ethyl, F, or deuterium; or, R 3 is H, methyl, or deuterium; (4) R 4 is ethyl, F or deuterium, or, R 4 is methyl or deuterium.

6. The anthracycline compound of claim 1, or a pharmaceutically acceptable salt thereof, having the formula: ###0002### wherein R is H, OH, or OCH3. which is selected from any one of the following schemes: Scheme 1: R 1 and R 2 independently are C 1-6 alkyl, or R 1 , R 2 and the C atom to which they are attached together form a cyclopropyl or cyclobutyl group; R 3 and R 4 independently are C 1-6 alkyl, or R 3 , R 4 and the C atom to which they are attached together form a cyclopropyl or cyclobutyl group; R 5 is C 1-6 alkyl; Scheme 2: R 1 and R 2 independently C 1-6 alkyl, R 3 and R 4 independently C 1-6 alkyl, R 5 C 1-6 alkyl; Scheme 3: R 1 is H or deuterium, R 2 is deuterium, R 3 is H or deuterium, R 4 is deuterium, R 5 is C 1-6 alkyl.

7. The anthracycline compound of claim 1, or a pharmaceutically acceptable salt thereof, having the formula: ###0002### wherein R is H, OH, or OCH3. The anthracycline compound as shown in Formula I is any one of the following compounds:

8. A method for preparing an anthracite compound as shown in Formula I, characterized in that, which comprises the following steps: In a solvent, in the presence of a deprotecting agent, the compound of formula I-1 undergoes a deprotection reaction as shown below to give an anthracycline of formula I, R 1 , R 2 , R 3 , R 4 , and R 5 are as defined in any one of claims 1-7, R a and R b are independently a hydroxyl protecting group, for example R a is a tert-butyldiphenylsilyl group, R b is an acetyl group.

9. The method for preparing anthracycline compounds as shown in Formula I according to claim 8, characterized in that, The compound of formula I-1 is a compound of formula I-1-X, the anthracycline of formula I is a compound of formula I-X; or, as shown in formula I-1, the compound is a compound as shown in formula I-1-Y, the anthracycline as shown in formula I is a compound as shown in formula I-Y; 10. The method for preparing anthracycline compounds as shown in Formula I according to claim 8, characterized in that, The preparation method of the anthracycline compound as shown in formula I further comprises a preparation method of a compound as shown in formula I-1, which comprises the following steps: performing an addition reaction of a compound as shown in formula I-A and a compound as shown in formula I-B as shown in the following formula in a solvent in the presence of an acid to obtain a compound as shown in formula I-1, R 1 , R 2 , R 3 , R 4 , and R 5 are as defined in any one of claims 1-6, R a and R b are independently a hydroxyl protecting group, for example R a is a t-butyldiphenylsilyl group, R b is an acetyl group; Preferably, the compound is of formula I-A 11. A compound of formula I-1 or of formula I-A, ###00010### I-1 I-A R 1 , R 2 , R 3 , R 4 and R 5 are as defined in any one of claims 1 to 6, R a and R b are independently a hydroxyl protecting group, for example R a is a tert-butyldiphenylsilyl group, R b is an acetyl group.

12. A compound according to claim 11 of formula I-1 or of formula I-A, wherein The compound as shown in formula I-1 is or, the compound of Formula I-A 13. The compound as shown in formula I-1 or formula I-A according to claim 11, wherein, The compound as shown in formula I-1 is any one of the following compounds: or, the compound of Formula I-A is any one of the following:

14. A pharmaceutical composition comprising: (1) an anthracene compound as shown in formula I or a pharmaceutically acceptable salt thereof according to any one of claims 1-7; and (2) a pharmaceutical adjuvant.

15. Use of an anthracene compound as shown in formula I or a pharmaceutically acceptable salt thereof according to any one of claims 1-7 or a pharmaceutical composition according to claim 14 in the preparation of a medicament for the prevention and / or treatment of a tumor, which can be malignant lymphoma, lung cancer, cervical cancer, ovarian cancer, breast cancer or Kaposi's sarcoma; the lung cancer can be non-small cell lung cancer.

Images