TWI796144B - Thienopyrimidinedione compounds and their applications - Google Patents

Abstract

The invention discloses a series of thienopyrimidine diketone compounds and their application used, specifically disclosed the compound represented by formula (II) and the pharmaceutically acceptable salt thereof.

Description

Thienopyrimidinedione compounds and their applications

[CROSS-REFERENCE TO RELATED APPLICATIONS]

The present invention claims the following priority: CN202110205200.0, the application date is February 23, 2021; CN202210128005.7, the application date is February 10, 2022.

The present invention relates to a series of thienopyrimidinedione compounds and applications thereof, specifically to compounds represented by formula (II) and pharmaceutically acceptable salts thereof.

GnRH (gonadotropin-releasing hormone, gonadotropin-releasing hormone) is a decapeptide hormone (pGlu-His-Trp-Ser-Tyr-Gly-Leu-Arg-Pro-Gly-NH2) synthesized by hypothalamic brain neuroendocrine cells, It is a central regulator in the endocrine nervous system. It reaches the pituitary gland through the hypothalamic pituitary portal circulatory system, binds to the GnRH receptor cells of the anterior pituitary gland, promotes the secretion of luteinizing hormone (LH) and follicle estrogen (FSH), and regulates the normal development of the ovary and corpus luteum. In the hypothalamus- The pituitary-gonadal axis plays an important role. LH and FSH bind to receptors on specific cells in the ovaries or testes, stimulating steroid production. The presence of steroids aggravates endometriosis, uterine fibroids, and prostate cancer, and it is necessary to give long-acting peptide GnRH receptor agonists or antagonists for control treatment.

Polypeptide GnRH receptor agonist or antagonist compounds have many problems, such as oral absorption, dosage form, dosage volume, drug stability, sustained action, and metabolic stability. Small molecular compounds can be administered orally, which is convenient and quick, and has obvious advantages. GnRH receptor antagonists bind GnRH receptors competitively, block GnRH binding to receptors, directly inhibit the hypothalamus-pituitary-ovarian axis, thereby inhibiting the secretion of follicle-stimulating hormone and luteinizing hormone, reducing estrogen levels, and acting Fast, with few side effects.

Although there are some meaningful clinical trials in these fields, such as Elagolix, Relugolix, Linzagolix and SKI2670, which are at the forefront of the development of small molecule GnRH receptor antagonists, more effective small molecule GnRH receptor antagonists still need to be researched and developed. body antagonists.

The invention provides a novel thienopyrimidine diketone GnRH receptor antagonist, and this type of structure has significant inhibitory activity on GnRH receptors.

其中， R ₁選自C _3-6環烷基、-S(=O) _m-C _1-3烷基、-S(=O) _m-C _3-6環烷基和-O-C _3-6環烷基，所述C _3-6環烷基、-S(=O) _m-C _1-3烷基、-S(=O) _m-C _3-6環烷基和-O-C _3-6環烷基分別獨立地任選被1、2或3個R _a取代； R ₂和R ₃各自獨立地選自H、F、Cl、Br、I和C _1-3烷基，所述C _1-3烷基任選被1、2或3個R _b取代； 各R ₄分別獨立地選自F、Cl、Br、I、C _1-3烷氧基、-O-C _3-6環烷基和C _3-6環烷基，所述C _1-3烷氧基、-O-C _3-6環烷基和C _3-6環烷基分別獨立地任選被1、2或3個R _c取代； 或者，R ₁和R ₂與它們連接的原子一起形成環B，環B選自5-6元雜環基，所述5-6元雜環基任選被1、2或3個F取代； R ₈選自-C(=O)OR ₆和-P(=O)(R ₇)OR ₆； R ₆選自H和C _1-3烷基，所述C _1-3烷基任選被1、2或3個F取代； R ₇選自C _1-3烷基，所述C _1-3烷基任選被1、2或3個F取代； n選自1、2、3和4； m選自0、1和2； 各R _a、R _b和R _c分別獨立地選自H、F、Cl、Br和I。 The present invention provides a compound represented by formula (II) or a pharmaceutically acceptable salt thereof,

Wherein, R ₁ is selected from C _3-6 cycloalkyl, -S(=O) _m -C _1-3 alkyl, -S(=O) _m -C _3-6 cycloalkyl and -OC _3-6 Cycloalkyl, the C _3-6 cycloalkyl, -S(=O) _m -C _1-3 alkyl, -S(=O) _m -C _3-6 cycloalkyl and -OC _3-6 Cycloalkyl is independently optionally substituted by 1, 2 or 3 R _a ; R ₂ and R ₃ are each independently selected from H, F, Cl, Br, I and C _1-3 alkyl, the C _{1 -3} alkyl is optionally substituted by 1, 2 or 3 R _b ; _each R is independently selected from F, Cl, Br, I, C _1-3 alkoxy, -OC _3-6 cycloalkyl and C _3-6 cycloalkyl, the C _1-3 alkoxy, -OC _3-6 cycloalkyl and C _3-6 cycloalkyl are independently optionally substituted by 1, 2 or 3 R _c ; Or, R ₁ and R ₂ form ring B together with the atoms they are connected to, and ring B is selected from 5-6 membered heterocyclic groups, and the 5-6 membered heterocyclic groups are optionally substituted by 1, 2 or 3 F; R ₈ is selected from -C(=O)OR ₆ and -P(=O)(R ₇ )OR ₆ ; R ₆ is selected from H and C _1-3 alkyl, the C _1-3 alkyl is optionally 1, 2 or 3 F substitutions; R ₇ is selected from C _1-3 alkyl, said C _1-3 alkyl is optionally substituted by 1, 2 or 3 F; n is selected from 1, 2, 3 and 4 m is selected from 0, 1 and 2; each of R _a , R _b and R _c is independently selected from H, F, Cl, Br and I.

In some schemes of the present invention, above-mentioned R ₁ is selected from cyclopropyl, cyclobutyl, -S(=O) _m -C _1-3 alkyl and -O-cyclopropyl, said cyclopropyl, cyclo Butyl, -S(=O) _m -C _1-3 alkyl and -O-cyclopropyl are independently optionally substituted by 1, 2 or 3 R _a , and other variables are as defined in the present invention.

In some schemes of the present invention, the above-mentioned R ₁ is selected from -S(=O) _m -C _1-3 alkyl, and the -S(=O) _m -C _1-3 alkyl is optionally replaced by 1, 2 or 3 R _a substitutions, other variables are as defined in the present invention.

和

，其他變數如本發明所定義。 在本發明的一些方案中，上述R ₁選自-SCH ₃、-S(=O)CH ₃、-S(=O) ₂CH ₃、

和

，其他變數如本發明所定義。 In some aspects of the present invention, the above R ₁ is selected from -SCH ₃ , -SCF ₃ , -SCF ₃ , -SCH ₂ CH ₃ , -SCH ₂ CF ₃ , -S(=O)CH ₃ , -S(= O) ₂ CH ₃ ,

and

, other variables are as defined in the present invention. In some schemes of the present invention, the above-mentioned R ₁ is selected from -SCH ₃ , -S(=O)CH ₃ , -S(=O) ₂ CH ₃ ,

and

, other variables are as defined in the present invention.

In some aspects of the present invention, the above R ₁ is selected from -SCH ₃ , -SCF ₃ , -SCF ₃ , -SCH ₂ CH ₃ , -SCH ₂ CF ₃ , -S(=O)CH ₃ and -S(= O) ₂ CH ₃ , other variables are as defined in the present invention.

In some aspects of the present invention, the above-mentioned R ₁ is selected from -SCH ₃ , -S(=O)CH ₃ and -S(=O) ₂ CH ₃ , and other variables are as defined in the present invention.

In some aspects of the present invention, the above-mentioned R ₂ is selected from H, F, Cl and CH ₃ , and the CH ₃ is optionally substituted by 1, 2 or 3 F, and other variables are as defined in the present invention.

In some aspects of the present invention, the above R ₃ is selected from H, F, Cl and CH ₃ , the CH ₃ is optionally substituted by 1, 2 or 3 F, and other variables are as defined in the present invention.

In some solutions of the present invention, the above R ₂ and R ₃ are each independently selected from H, and other variables are as defined in the present invention.

In some schemes of the present invention, the above-mentioned R ₄ is selected from F, Cl, -OCH ₃ , -OCH ₂ CH ₃ , -O-cyclopropyl, -O-cyclobutyl, cyclopropyl and cyclobutyl, so The above -OCH ₃ , -OCH ₂ CH ₃ , -O-cyclopropyl, -O-cyclobutyl, cyclopropyl and cyclobutyl are independently optionally substituted by 1, 2 or 3 F, other variables such as defined in the present invention.

、

、環丙基和環丁基，其他變數如本發明所定義。 In some schemes of the present invention, the above-mentioned R ₄ is selected from F, Cl, -OCH ₃ , -OCH ₂ CH ₃ ,

,

, cyclopropyl and cyclobutyl, other variables are as defined herein.

、F和-OCH ₃，其他變數如本發明所定義。 In some schemes of the present invention, above-mentioned R ₄ is selected from

, F and -OCH ₃ , other variables are as defined in the present invention.

In some solutions of the present invention, the above-mentioned R ₄ is selected from F and -OCH ₃ , and other variables are as defined in the present invention.

選自

和

，其他變數如本發明所定義。 In some solutions of the present invention, the above-mentioned structural units

selected from

and

, other variables are as defined in the present invention.

選自

，其他變數如本發明所定義。 In some solutions of the present invention, the above-mentioned structural units

selected from

, other variables are as defined in the present invention.

、

、

和

，所述

、

、

和

分別獨立地任選被1、2或3個F取代，其他變數如本發明所定義。 In some solutions of the present invention, the above-mentioned R ₁ and R ₂ are linked together to form

,

,

and

, the

,

,

and

are independently optionally substituted by 1, 2 or 3 Fs, and other variables are as defined in the present invention.

和

，其他變數如本發明所定義。 In some solutions of the present invention, the above-mentioned R ₁ and R ₂ are linked together to form

and

, other variables are as defined in the present invention.

、

和

，其他變數如本發明所定義。 In some solutions of the present invention, the above-mentioned R ₁ and R ₂ are linked together to form

,

and

, other variables are as defined in the present invention.

，其他變數如本發明所定義。 In some solutions of the present invention, the above-mentioned R ₁ and R ₂ are linked together to form

, other variables are as defined in the present invention.

In some solutions of the present invention, the above-mentioned R ₈ is selected from -C(=O)OH, -C(=O)OCH ₃ , -C(=O)OCH ₂ CH ₃ , -C(=O)OC(CH ₃ ) ₂ , -P(=O)(CH ₃ )OH and -P(=O)(CH ₃ )OCH ₃ , other variables are as defined herein.

In some solutions of the present invention, the above-mentioned R ₈ is selected from -C(=O)OH and -P(=O)(CH ₃ )OH, and other variables are as defined in the present invention.

In some solutions of the present invention, the above-mentioned R ₈ is selected from -C(=O)OH, and other variables are as defined in the present invention.

， 其中，R ₁、R ₂、R ₃、R ₄和n如本發明所定義。 In some aspects of the present invention, the above-mentioned compound or a pharmaceutically acceptable salt thereof is selected from the group consisting of,

, wherein, R ₁ , R ₂ , R ₃ , R ₄ and n are as defined in the present invention.

、

和

， 其中， 結構單元

選自

、

和

； R ₅選自C _1-3烷基，所述C _1-3烷基任選被1、2或3個R _a取代； 環A選自C _3-6環烷基，所述C _3-6環烷基任選被1、2或3個R _a取代； p選自0和1； 環B、R ₂、R ₃、R ₄、n和R _a如本發明所定義。 In some aspects of the present invention, the above-mentioned compound or a pharmaceutically acceptable salt thereof is selected from the group consisting of,

,

and

, where the structural unit

selected from

,

and

; R ₅ is selected from C _1-3 alkyl, the C _1-3 alkyl is optionally substituted by 1, 2 or 3 R _a ; Ring A is selected from C _3-6 cycloalkyl, the C _{3- 6} cycloalkyl is optionally substituted by 1, 2 or 3 R _a ; p is selected from 0 and 1; ring B, R ₂ , R ₃ , R ₄ , n and R _a are as defined in the present invention.

、

和

， 其中，

為不存在或雙鍵； R ₅選自C _1-3烷基，所述C _1-3烷基任選被1、2或3個R _a取代； 環A選自C _3-6環烷基，所述C _3-6環烷基任選被1、2或3個R _a取代； p選自0和1； 環B、R ₂、R ₃、R ₄、m、n和R _a如本發明所定義。 In some aspects of the present invention, the above-mentioned compound or a pharmaceutically acceptable salt thereof is selected from the group consisting of,

,

and

, in,

For absence or double bond; R ₅ is selected from C _1-3 alkyl, the C _1-3 alkyl is optionally substituted by 1, 2 or 3 R _a ; ring A is selected from C _3-6 cycloalkyl , the C _3-6 cycloalkyl group is optionally substituted by 1, 2 or 3 R _a ; p is selected from 0 and 1; ring B, R ₂ , R ₃ , R ₄ , m, n and R _a are as present invention defined.

其中， R ₁選自C _3-6環烷基、-S(=O) _m-C _1-3烷基、-S(=O) _m-C _3-6環烷基和-O-C _3-6環烷基，所述C _3-6環烷基、-S(=O) _m-C _1-3烷基、-S(=O) _m-C _3-6環烷基和-O-C _3-6環烷基任選被1、2或3個R _a取代； R ₂和R ₃各自獨立地選自H、F、Cl、Br、I和C _1-3烷基，所述C _1-3烷基任選被1、2或3個R _b取代； 各R ₄分別獨立地選自F、Cl、Br、I、C _1-3烷氧基、-O-C _3-6環烷基和C _3-6環烷基，所述C _1-3烷氧基、-O-C _3-6環烷基和C _3-6環烷基任選被1、2或3個R _c取代； 或者，R ₁和R ₂連接在一起形成環B，其中環B選自

、

和

； n選自1、2、3和4； m選自0、1和2； 各R _a、R _b和R _c分別獨立地選自H、F、Cl、Br和I。 The present invention also provides a compound represented by formula (I) or a pharmaceutically acceptable salt thereof,

Wherein, R ₁ is selected from C _3-6 cycloalkyl, -S(=O) _m -C _1-3 alkyl, -S(=O) _m -C _3-6 cycloalkyl and -OC _3-6 Cycloalkyl, the C _3-6 cycloalkyl, -S(=O) _m -C _1-3 alkyl, -S(=O) _m -C _3-6 cycloalkyl and -OC _3-6 Cycloalkyl is optionally _substituted by 1, ₂ or ₃ R; R and R are each independently selected from H, F, Cl, Br, I and C _1-3 alkyl, said C _1-3 alkane The group is optionally substituted by 1, 2 or 3 R _b ; each R ₄ is independently selected from F, Cl, Br, I, C _1-3 alkoxy, -OC _3-6 cycloalkyl and C _{3- 6} cycloalkyl, said C _1-3 alkoxy, -OC _3-6 cycloalkyl and C _3-6 cycloalkyl are optionally substituted by 1, 2 or 3 R _c ; or, R ₁ and R ₂ linked together to form ring B, wherein ring B is selected from

,

and

n is selected from 1, 2, 3 and 4; m is selected from 0, 1 and 2; each of R _a , R _b and R _c is independently selected from H, F, Cl, Br and I.

Some schemes of the present invention are formed by any combination of the above-mentioned variables.

、

、

、

、

、

、

、

、

、

、

、

、

和

。 The present invention also provides the following compounds or pharmaceutically acceptable salts thereof,

,

,

,

,

,

,

,

,

,

,

,

,

and

.

The present invention also provides the use of the above compound or a pharmaceutically acceptable salt thereof in the preparation of drugs related to GnRH receptor antagonists.

In some solutions of the present invention, the above-mentioned GnRH receptor antagonist-related drugs are drugs for preventing and/or treating endometriosis and/or uterine fibroid-related diseases.

In some aspects of the present invention, the above-mentioned drugs related to GnRH receptor antagonists are drugs for preventing and/or treating diseases related to prostate cancer.

The present invention also provides the following in vivo drug efficacy test methods: Experimental purpose: the drug effect evaluation of the compound of the present invention in the mouse endometriosis model. Test plan: 2.1 Main reagents and consumables C57BL6/J female mice, caliper (ARZ-1331), 8-0 suture, stereomicroscope. 2.2 Experimental steps 8-week-old female C57BL6/J female mouse animal estrous cycle detection: vaginal smear observation, since the entire estrous cycle is about 4 days, according to the actual smear results, select mice that meet the specific estrous cycle on the day of the experiment for the experiment.

Modeling: Uterine horns were excised from donor mice in diestrus, opened longitudinally and the excised biopsies cut to a size of 2 x 2 mm. Recipient mice were anesthetized with isoflurane gas and a 1 cm incision was made along the midline to expose the abdominal cavity. Mice with endometriosis: Four uterine fragments from a donor were sutured to the peritoneal wall of mice in estrus; Sham-operated mice: Fragments of abdominal fat of similar size were sutured. Use 8-0 black silk sutures to suture the grafted tissue as well as the abdominal muscle and skin. Sham group (sham operation group): each sham group opened the abdominal cavity to transplant fat fragments, and the rest of the steps were the same.

After 4 weeks of modeling, a second laparotomy was performed, and the volume of the ectopic lesion (V ₁ ) was measured and calculated using a vernier caliper to determine the success of the modeling. Twelve mice with relatively uniform V ₁ were selected and divided into 2 groups, 6 mice in each group: Vehicle (vehicle control group) and drug group.

Sham group (sham operation group, 6 rats): normal feeding; Vehicle group (vehicle control group, 6 rats): intragastric administration of vehicle once a day for 8 consecutive weeks. The vehicle was 10% DMAC+10%solutol+80% normal saline; Compound group (6 rats): gavage 100 mpk at 10:00 am every day for 8 consecutive weeks, the vehicle is 10% DMAC + 10% solutol + 80% normal saline.

。 技術效果 After 8 weeks of administration, the mice were taken: use a vernier caliper to measure and calculate the volume of ectopic focus (V ₂ ), and calculate the inhibition rate. Calculation formula: ectopic lesion volume V = π/6×length×width×height, length means the length of ectopic lesion, width means the width of ectopic lesion, height means the height of ectopic lesion;

. technical effect

The compound of the present invention has a significant inhibitory effect on human gonadotropin-releasing hormone receptors, and has a significant anti-tumor effect on the human prostate cancer LNCaP clone FGC xenograft tumor model, and the PK results show the exposure of the compound of the present invention in plasma High, low clearance, long half-life, high oral bioavailability, exhibits excellent pharmacokinetic properties, and is a good molecule for oral administration. Definition and Description

Unless otherwise stated, the following terms and phrases used herein are intended to have the following meanings. A specific term or phrase should not be considered indeterminate or unclear if it is not specifically defined, but should be understood according to its ordinary meaning. When a trade name appears herein, it is intended to refer to its corresponding trade name or its active ingredient.

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

The term "pharmaceutically acceptable salt" refers to a salt of a compound of the present invention, which is prepared from a compound having a specific substituent found in the present invention and a relatively non-toxic acid or base. When compounds of the present invention contain relatively acidic functional groups, base addition salts can be obtained by contacting such compounds with a sufficient amount of base, either neat solution or in a suitable inert solvent. Pharmaceutically acceptable base addition salts include sodium, potassium, calcium, ammonium, organic amine or magnesium salts or similar salts. When compounds of the present invention contain relatively basic functionalities, acid addition salts can be obtained by contacting such compounds with a sufficient amount of the acid, either neat solution or in a suitable inert solvent. Examples of pharmaceutically acceptable acid addition salts include salts of inorganic acids including, for example, hydrochloric acid, hydrobromic acid, nitric acid, carbonic acid, bicarbonate, phosphoric acid, monohydrogenphosphate, dihydrogenphosphate, sulfuric acid, Hydrogen sulfate, hydriodic acid, phosphorous acid, etc.; and organic acid salts, such as acetic acid, propionic acid, isobutyric acid, maleic acid, malonic acid, benzoic acid, succinic acid, suberic acid, Fumaric acid, lactic acid, mandelic acid, phthalic acid, benzenesulfonic acid, p-toluenesulfonic acid, citric acid, tartaric acid and methanesulfonic acid and similar acids; also salts of amino acids such as arginine, etc. , and salts of organic acids such as glucuronic acid. Certain specific compounds of the present invention contain basic and acidic functional groups and can thus be converted into either base or acid addition salts.

The pharmaceutically acceptable salts of the present invention can be synthesized from the parent compound containing acid groups or bases by conventional chemical methods. In general, such salts are prepared by reacting the free acid or base form of these compounds with a stoichiometric amount of the appropriate base or acid in water or an organic solvent or a mixture of both.

The compounds of the invention may exist in particular geometric or stereoisomeric forms. The present invention contemplates all such compounds, including cis and trans isomers, (-)- and (+)-enantiomers, ( R )- and ( S )-enantiomers, diastereomers isomers, ( D )-isomers, ( L )-isomers, and their racemic and other mixtures, such as enantiomerically or diastereomerically enriched mixtures, all of which are included in this within the scope of the invention. Additional asymmetric carbon atoms may be present in substituents such as alkyl groups. All such isomers, as well as mixtures thereof, are included within the scope of the present invention.

Unless otherwise stated, the terms "enantiomer" or "optical isomer" refer to stereoisomers that are mirror images of each other.

Unless otherwise stated, the terms "cis-trans isomers" or "geometric isomers" arise from the inability to rotate freely due to the double bond or the single bond of the carbon atoms forming the ring.

Unless otherwise indicated, the term "diastereoisomer" refers to stereoisomers whose molecules have two or more chiral centers and which are not mirror images of the molecules.

Unless otherwise stated, the term "tautomer" or "tautomeric form" means that isomers with different functional groups are in dynamic equilibrium at room temperature and are rapidly interconvertible. If tautomerism is possible (eg, in solution), then chemical equilibrium of the tautomers can be achieved. For example, proton tautomers (also called prototropic tautomers) include interconversions via migration of a proton, such as keto-enol isomerization and imine-enamine isomerization. structured. Valence isomers (valence tautomers) Interconversions involving recombination of some of the bonding electrons. A specific example of keto-enol tautomerization is the interconversion between two tautomers of pentane-2,4-dione and 4-hydroxypent-3-en-2-one.

Unless otherwise stated, the terms "enriched in an isomer", "enriched in an isomer", "enriched in an enantiomer" or "enantiomerically enriched" refer to one of the isomers or enantiomers The content is less than 100%, and the content of the isomer or enantiomer is greater than or equal to 60%, or greater than or equal to 70%, or greater than or equal to 80%, or greater than or equal to 90%, or greater than or equal to 95%, or greater than or equal to 96% %, or greater than or equal to 97%, or greater than or equal to 98%, or greater than or equal to 99%, or greater than or equal to 99.5%, or greater than or equal to 99.6%, or greater than or equal to 99.7%, or greater than or equal to 99.8%, or greater than or equal to 99.9%.

Unless otherwise stated, the terms "isomer excess" or "enantiomeric excess" refer to the difference between the relative percentages of two isomers or two enantiomers. For example, if the content of one isomer or enantiomer is 90% and the other isomer or enantiomer is 10%, then the isomer or enantiomeric excess (ee value) is 80%.

The compounds of the present invention may contain unnatural proportions of atomic isotopes at one or more of the atoms that constitute the compounds. For example, compounds may be labeled with radioactive isotopes such as tritium ( ^3H ), iodine-125 ( ^125I ) or C-14 ( ^14C ). For another example, heavy hydrogen can be used to replace hydrogen to form deuterated drugs. The bond formed by deuterium and carbon is stronger than the bond formed by ordinary hydrogen and carbon. Compared with non-deuterated drugs, deuterated drugs can reduce toxic side effects and increase drug stability. , enhance the efficacy, prolong the biological half-life of drugs and other advantages. All changes in isotopic composition of the compounds of the invention, whether radioactive or not, are included within the scope of the invention.

The term "optional" or "optionally" means that the subsequently described event or circumstance can but need not occur, and that the description includes instances where said event or circumstance occurs and instances where said event or circumstance does not occur .

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

When any variable (such as R) occurs more than once in the composition or structure of a compound, its definition is independent in each case. Thus, for example, if a group is substituted with 0-2 R, said group may optionally be substituted with up to two R, with independent options for each occurrence of R. Also, combinations of substituents and/or variations thereof are permissible only if such combinations result in stable compounds.

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

When one of the variables is selected from a single bond, it means that the two groups connected are directly connected. For example, when L in A-L-Z represents a single bond, it means that the structure is actually A-Z.

）、直形虛線鍵（

）、或波浪線（

）表示。例如-OCH ₃中的直形實線鍵表示通過該基團中的氧原子與其他基團相連；

中的直形虛線鍵表示通過該基團中的氮原子的兩端與其他基團相連；

中的波浪線表示通過該苯基基團中的1和2位碳原子與其他基團相連；

表示該呱啶基上的任意可連接位點可以通過1個化學鍵與其他基團相連，至少包括

、

、

、

這4種連接方式，即使-N-上畫出了H原子，但是

仍包括

這種連接方式的基團，只是在連接1個化學鍵時，該位點的的H會對應減少1個變成相應的一價呱啶基。 Unless otherwise specified, when a group has one or more linkable sites, any one or more sites of the group can be linked to other groups through chemical bonds. When the connection method of the chemical bond is not positioned, and there is an H atom at the connectable site, when the chemical bond is connected, the number of H atoms at the site will decrease correspondingly with the number of chemical bonds connected to become the corresponding valence group. The chemical bond that described site is connected with other groups can use straight solid line bond (

), straight dashed key (

), or the squiggly line (

)express. For example, the straight-shaped solid-line bond in _-OCH3 indicates that it is connected to other groups through the oxygen atom in the group;

The straight dotted line bond in indicates that the two ends of the nitrogen atom in the group are connected to other groups;

The wavy lines in indicate that the 1 and 2 carbon atoms in the phenyl group are connected to other groups;

Indicates that any connectable site on the piperidyl group can be connected to other groups through a chemical bond, including at least

,

,

,

These 4 connection methods, even if the H atom is drawn on -N-, but

still include

For groups with this connection method, only when a chemical bond is connected, the H at this site will be reduced by one to become the corresponding monovalent piperidinyl group.

” 表示不存在或雙鍵，例如

表示

、

或

。 Unless otherwise specified, the term "

” means absence or double bond, e.g.

express

,

or

.

Unless otherwise specified, the term "C _1-3 alkyl" by itself or in combination with other terms means a linear or branched saturated hydrocarbon group consisting of 1 to 3 carbon atoms, respectively. The C _1-3 alkyl includes C _1-2 and C _2-3 alkyl, etc.; it can be monovalent (such as methyl), divalent (such as methylene) or multivalent (such as methine) . Examples of C _1-3 alkyl include, but are not limited to, methyl (Me), ethyl (Et), propyl (including n -propyl and isopropyl) and the like.

Unless otherwise specified, the term "C _1-3 alkoxy" by itself or in combination with other terms respectively means those alkyl groups containing 1 to 3 carbon atoms attached to the rest of the molecule through an oxygen atom. The C _1-3 alkoxy group includes C _1-2 , C _2-3 , C ₃ and C ₂ alkoxy groups and the like. Examples of C _1-3 alkoxy include, but are not limited to, methoxy, ethoxy, propoxy (including n-propoxy and isopropoxy), and the like.

Unless otherwise specified, "C _3-6 cycloalkyl" means a saturated cyclic hydrocarbon group composed of 3 to 6 carbon atoms, which is a monocyclic and bicyclic system, and the C _3-6 cycloalkyl includes C _3-5 , C _4-5 and C _5-6 cycloalkyl, etc.; it may be monovalent, divalent or multivalent. Examples of C _3-6 cycloalkyl include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl and the like.

、

、

、

或

。 Unless otherwise specified, the term "5-6 membered heterocyclyl" by itself or in combination with other terms represents a saturated or unsaturated monocyclic group consisting of 5 or 6 ring atoms, 1, 2, 3 or 4 3 ring atoms are heteroatoms independently selected from O, S, and N, and the remainder are carbon atoms, wherein the nitrogen atom is optionally quaternized, and the nitrogen and sulfur heteroatoms may be optionally oxidized (i.e., NO and S(O) _p , p is 1 or 2). In addition, with respect to the "5-6 membered heterocyclic group", a heteroatom may occupy the attachment position of the heterocycloalkyl group to the rest of the molecule. The 5-6 membered heterocyclic group includes 5-membered and 6-membered heterocycloalkyl, heterocycloalkenyl and heteroaryl. Examples of 5-6 membered heterocyclic groups include, but are not limited to, pyrrolyl, pyrazolyl, imidazolyl, pyrrolidinyl, pyrazolidinyl, imidazolidinyl, tetrahydrothiophenyl (including tetrahydrothiophen-2-yl and Tetrahydrothiophen-3-yl, etc.), tetrahydrofuryl (including tetrahydrofuran-2-yl, etc.), tetrahydropyranyl, piperidyl (including 1-piperidyl, 2-piperidyl and 3-piperidyl etc.), piperazinyl (including 1-piperazinyl and 2-piperazinyl, etc.), morpholinyl (including 3-morpholinyl and 4-morpholinyl, etc.), dioxanyl, dithianyl , isoxazolidinyl, isothiazolidinyl, 1,2-oxazinyl, 1,2-thiazinyl, hexahydropyridazinyl,

,

,

,

or

.

表示離域π電子雲。例如，結構式

、

和

所表示的結構均相同，結構式

、

、

和

所表示的結構均相同。其可以是單環或稠合多環體系，其中各個環均為芳香性的。除非另有規定，該環任選地包含0、1或多個獨立選自O、S和N的雜原子。 Unless otherwise specified, the term "aromatic ring" is a cyclic group having a conjugated π-electron system covered by a cloud of delocalized π-electrons between its atoms. In the structural formula, when the atomic valence state and covalent bond formation rules are met, it can be written in the form of alternating single and double bonds, or it can be written in the form of

represents a delocalized π-electron cloud. For example, the formula

,

and

The structures represented are the same, the structural formula

,

,

and

The structures shown are all the same. It can be a single ring or a fused polycyclic ring system in which each ring is aromatic. Unless otherwise specified, the ring optionally contains 0, 1 or more heteroatoms independently selected from O, S and N.

Unless otherwise specified, C _n-n+m or C _n -C _n+m includes any specific instance of n to n+m carbons, e.g. C _1-12 includes C ₁ , C ₂ , C ₃ , C ₄ , C ₅ , C ₆ , C ₇ , C ₈ , C ₉ , C ₁₀ , C ₁₁ , and C ₁₂ , also including any range from n to n+m, for example, C _1-12 includes C _1-3 , C _1-6 , C _1-9 , C _3-6 , C _3-9 , C _3-12 , C _6-9 , C _6-12 , and C _9-12 etc.; similarly, n to n+ m-membered means that the number of atoms on the ring is from n to n+m. For example, a 3-12-membered ring includes a 3-membered ring, a 4-membered ring, a 5-membered ring, a 6-membered ring, a 7-membered ring, an 8-membered ring, and a 9-membered ring. 10-membered rings, 11-membered rings, and 12-membered rings, including any range from n to n+m, for example, 3-12-membered rings include 3-6-membered rings, 3-9-membered rings, and 5-6-membered rings , 5-7-membered ring, 6-7-membered ring, 6-8-membered ring, and 6-10-membered ring, etc.

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, the embodiments formed by combining them with other chemical synthesis methods, and equivalents well known to those skilled in the art Alternatively, preferred embodiments include, but are not limited to, the examples of the present invention.

The structure of the compounds of the present invention can be confirmed by conventional methods known to those skilled in the art. If the absolute configuration of the compound in the present invention is related, the absolute configuration can be confirmed by conventional technical means in the art. For example, single crystal X-ray diffraction (SXRD), the cultivated single crystal is collected with Bruker D8 venture diffractometer to collect diffraction intensity data, the light source is CuKα radiation, scanning method: φ/ω scanning, after collecting relevant data, further adopt direct method (Shelxs97) By analyzing the crystal structure, the absolute configuration can be confirmed.

The following acronyms are used in the present invention: DMSO stands for dimethylsulfoxide; MeOH stands for methanol; ACN stands for acetonitrile; DEA stands for diethylamine; _CO2 stands for carbon dioxide; DMAC stands for N,N-dimethylacetamide; Solutol Represents polyethylene glycol-15 hydroxystearate; psi represents pounds force per square inch; PEG represents polyethylene glycol.

The solvent used in the present invention is commercially available. Compounds were named according to conventional naming principles in the field or using ChemDraw® software, and commercially available compounds were named in suppliers’ catalogs.

The present invention will be described in detail through examples below, but it does not imply any unfavorable limitation to the present invention. The present invention has been described in detail herein, and its specific embodiments have also been disclosed. For those skilled in the art, various changes and improvements will be made to the specific embodiments of the present invention without departing from the spirit and scope of the present invention. It is obvious.

Example 1

first step

To a solution of compound 1a (dimethylmaleate, 50 g, 346.92 mmol) and methyl thioglycolate (37.01 g, 348.68 mmol) in THF (300 mL) was added piperidine (886.18 mg, 10.41 mmol), and the mixture was stirred at 25°C for 2 hours. Added 300 ml of water, extracted twice with ethyl acetate, 200 ml each time, the combined organic phase was washed with 200 ml of saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to dryness to obtain compound 1b .

¹ H NMR (400 MHz, CDCl ₃ ) δ = 3.89 - 3.81 (m, 1H), 3.76 (s, 3H), 3.75 (d, 3H), 3.69 (s, 3H), 3.55 - 3.32 (m, 2H) , 3.00 (m, 1H), 2.73 (m, 1H).

second step

Add sodium (2.78 g, 120.92 mmol) to methanol (10 ml) below 20 °C, stir until the solid dissolves below 64 °C, configure sodium methoxide methanol solution, cool to 20 °C, add to 1b (10 g , 39.96 mmol) in tetrahydrofuran (20 ml), and the reaction solution was stirred at 66°C for 3 hours under nitrogen protection. Add isopropyl ether (100 ml) and acetic acid (1 ml), cool to 20 °C, filter, add the filter cake to a mixture of phosphoric acid (10 ml) and water (20 ml), extract twice with ethyl acetate, each 20 ml, the combined organic phase was washed with 30 ml of saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to dryness to obtain compound 1c .

third step

A mixture of compound 1c (7.4 g, 33.91 mmol), pyridine (4.03 g, 50.89 mmol), hydroxylamine hydrochloride (2.47 g, 35.57 mmol) was stirred at 50°C for 2 hours. Add 2 ml of phosphoric acid and 20 ml of water to the reaction solution, extract twice with ethyl acetate, 20 ml each time, combine the organic layers, wash with 20 ml of saturated sodium bicarbonate, wash once with 20 ml of saturated aqueous sodium chloride, and dry over anhydrous sodium sulfate , filtered, and the filtrate was concentrated to dryness under reduced pressure to obtain compound 1d .

the fourth step

To compound 1d (5.2 g, 22.29 mmol) in acetic acid (5 ml), 4 mol/L of ethyl acetate hydrochloride (52.00 ml) was added, and the reaction solution was stirred at 25°C for 12 hours. The reaction solution was filtered, and the filter cake was dried under reduced pressure to obtain the hydrochloride of compound 1e .

MS-ESI calculated [M+H-MeOH] ⁺ : 184.0, found 184.1.

the fifth step

Add potassium carbonate (1.65 g, 11.92 mmol) to compound 1e (2 g, 7.95 mmol, hydrochloride) in tetrahydrofuran (20 ml) and water (10 ml), dropwise at 5-10 °C Phenyl chloroformate (2.49 g, 15.89 mmol), and the reaction solution was stirred at 5-10°C for 1 hour. Add 50 ml of water to the reaction solution, extract twice with ethyl acetate, 50 ml each time, combine the organic phases, wash with 50 ml of saturated brine, dry over anhydrous sodium sulfate, filter, concentrate the filtrate to dryness, add ethyl acetate (5 ml ), petroleum ether (50 ml), beat and stir at 30°C for 30 minutes, filter, and dry the filter cake under reduced pressure to obtain compound 1f .

MS-ESI calculated [M+H] ⁺ : 336.1, found 336.1.

step six

To a solution of compound 1 g (2,4-difluoro-5-nitrophenol, 2 g, 11.42 mmol) in N,N-dimethylformamide (10 mL) at 27 °C, batchwise Potassium carbonate (3.16 g, 22.85 mmol) was added one at a time. Sodium methylthiolate (880.68 mg, 12.56 mmol) was then added to the reaction, which was stirred at 27°C for 2 hours. 2 mol/L hydrochloric acid was added to the reaction system to adjust the pH to 6. Water (30 mL) was added to the reaction system, extracted with ethyl acetate (30 mL×3), and the combined organic phases were washed with water (10 mL×6). The collected organic phase was dried with anhydrous sodium sulfate, filtered, and the filtrate was concentrated to obtain compound 1h . ¹ H NMR (400 MHz, DMSO- d ₆ ) δ = 10.87 (s, 1H), 7.43 (d, J = 6.6 Hz, 1H), 7.22 (d, J = 12.4 Hz, 1H), 2.49 (br s, 3H).

step seven

To a solution of compound 3-chloromethyl-1,2-difluoro-4-methoxybenzene (454.96 mg, 2.3 mmol) and compound 1h (400 mg, 1.97 mmol) in toluene (3 mL) Tetrabutylammonium bromide (63.46 mg, 196.86 micromoles) was added. A solution of sodium hydroxide (118.12 mg, 2.95 mmol) in water (0.3 ml) was then added to the reaction. The mixture was stirred at 50°C for 12 hours. The reaction solution was poured into 30 ml of water, extracted with ethyl acetate (20 ml × 3), and the combined organic phases were washed with water (10 ml × 6). The collected organic phase was dried with anhydrous sodium sulfate, filtered, the filtrate was concentrated, and purified by column chromatography (silica gel, petroleum ether: ethyl acetate = 1:0-20:1) to obtain compound 1i .

¹ H NMR (400 MHz, CDCl ₃ ) δ = 7.81 (d, J = 6.4 Hz, 1H), 7.16 (q, J = 9.2 Hz, 1H), 6.90 (d, J = 11.6 Hz, 1H), 6.70 - 6.62 (m, 1H), 5.30 (d, J = 1.2 Hz, 2H), 3.94 - 3.89 (m, 3H), 2.44 (s, 3H).

eighth step

To a solution of compound 1i (163 mg, 453.63 μM) in ethanol (5 mL) and water (0.5 mL), iron powder (126.88 mg, 2.27 mmol) and ammonium chloride (14.56 mg, 272.18 μM Ear). The mixture was stirred at 85°C for 1 hour. The reaction solution was filtered with diatomaceous earth, the filter cake was washed with dichloromethane (10 ml × 5), then water (10 ml) was added to the filtrate, extracted with dichloromethane (10 ml × 3), and the organic phases were combined , washed with water (10 ml × 2), washed with brine (10 ml × 2), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to obtain compound 1j .

MS-ESI calculated [M+H] ⁺ : 330.1, found 330.1.

Ninth step

To a solution of compound 1j (117 mg, 355.26 micromol) in THF (4 mL), compound 1f (119.13 mg, 355.26 micromol) and triethylamine (35.95 mg, 355.26 micromol) were added. The mixture was stirred at 70°C for 10 hours. The reaction system was cooled down to 27° C., and compound 1f (65 mg, 193.84 micromole) and triethylamine (35.95 mg, 355.26 micromole) were added to the reaction system. The mixture was stirred at 70°C for 5 hours. The reaction solution was poured into 30 ml of water, extracted with ethyl acetate (50 ml × 5), the combined organic phases were washed with water (20 ml × 5), washed with saturated brine (20 ml), and dried over anhydrous sodium sulfate, After filtration, the filtrate was concentrated and the crude product was purified by preparative chromatography (silica gel, petroleum ether: ethyl acetate = 2:1) to obtain compound 1k .

MS-ESI calculated [M+H] ⁺ : 571.1, found 571.1.

tenth step

To a solution of compound 1k (100 mg, 175.27 micromol) in tetrahydrofuran (1 ml) and methanol (1 ml) was added an aqueous solution (0.5 ml) of lithium hydroxide monohydrate (44.13 mg, 175.27 micromol), the The mixture was stirred at 28°C for 1 hour. Add 1 mol/L of dilute hydrochloric acid to the reaction solution to adjust the pH to about 6, then extract with ethyl acetate (5 mL×5), combine the organic phases and wash with water (5 mL×3), collect the organic phases with anhydrous Dry over sodium sulfate, filter, concentrate the filtrate, and purify the crude product by preparative chromatography (silica gel, dichloromethane:methanol=10:1) to obtain compound 1 .

MS-ESI calculated [M+H] ⁺ : 525.0, found 525.0.

¹ H NMR (400 MHz, DMSO- d ₆ ) δ = 11.98 (br s, 1H), 7.56 - 7.40 (m, 1H), 7.38 - 7.26 (m, 2H), 7.18 (d, J = 10.4 Hz, 1H ), 6.92 (br d, J = 9.2 Hz, 1H), 5.03 (s, 2H), 3.82 (s, 3H), 2.40 (s, 3H).

Examples 2 and 3

first step

To a solution of compound 1k (165 mg, 289.19 micromole) in THF (1 mL) and methanol (1 mL), sodium methoxide (31.24 mg, 578.38 micromole) was added, and the reaction was stirred at 28°C for 0.5 hours . Acetic acid (2 ml) and water (10 ml) were added to the reaction system, extracted with ethyl acetate (10 ml×2), and the combined organic phases were washed with brine (10 ml). The collected organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to obtain compound 2a .

MS-ESI calculated [M+H] ⁺ : 539.1, found 539.1.

¹ H NMR (400 MHz, CDCl ₃ ) δ = 10.34 (s, 1H), 7.12 (q, J = 9.2 Hz, 1H), 6.98 (d, J = 10.0 Hz, 1H), 6.93 (d, J = 6.4 Hz, 1H), 6.87 (s, 1H), 6.64 - 6.57 (m, 1H), 5.13 (dd, J = 1.6, 4.4 Hz, 2H), 3.96 (s, 3H), 3.81 (s, 3H), 2.39 (s, 3H).

second step

Potassium monopersulfate (251.15 mg, 408.53 micromole) was added to water (2 ml), and stirred at 28°C until potassium monopersulfate was completely dissolved. Then, a solution of compound 2a (88 mg, 163.41 micromole) in acetonitrile (1 mL) was added dropwise to the reaction system under rapid stirring. The mixture was stirred at 28°C for 18.5 hours. The reaction system was poured into water (10 ml). The aqueous phase was extracted with ethyl acetate (10 ml × 5), the combined organic phase was washed with water (10 ml × 5), washed with brine (10 ml), the collected organic phase was dried over anhydrous sodium sulfate, filtered, the filtrate was concentrated and the crude product Purification by preparative chromatography (silica gel, dichloromethane:methanol = 15:1) gave compounds 2b and 2c .

Compound 2b : MS-ESI calculated value [M+H] ⁺ : 571.0, found value 571.1; Compound 2c : MS-ESI calculated value [M+H] ⁺ : 555.0, found value 555.1.

third step

To a solution of compound 2b (23 mg, 40.31 micromol) in tetrahydrofuran (1 ml) and methanol (1 ml) was added an aqueous solution (0.5 ml) of sodium hydroxide (4.84 mg, 120.94 micromol), and the mixture was Stir at 28°C for 1.5 hours. Add 1 mol/L dilute hydrochloric acid to the reaction solution to adjust the pH to about 6, then extract with ethyl acetate (5 ml × 5), combine the organic phases and wash with water (5 ml × 3), collect the organic phases with anhydrous sulfuric acid Sodium-dried, filtered, and the filtrate was concentrated. The crude product was purified by preparative chromatography (silica gel, dichloromethane:methanol=10:1), and then separated by high-performance liquid chromatography (column: 3_Phenomenex Luna C18 75×30 mm*3 μm; Mobile phase: [water (0.05% hydrochloric acid)-acetonitrile]; acetonitrile%: 37%-57%) to obtain compound 2 .

MS-ESI calculated [M+H] ⁺ : 557.0, found 557.0.

¹ H NMR (400 MHz, DMSO- d ₆ ) δ = 12.10 (br s, 1H), 7.84 (d, J = 5.2 Hz, 1H), 7.80 (d, J = 8.4 Hz, 1H), 7.53 (q, J = 9.6 Hz, 1H), 7.34 (br s, 1H), 6.96 (br d, J = 9.4 Hz, 1H), 5.19 (s, 2H), 3.82 (s, 3H), 3.26 - 3.24 (m, 3H ).

the fourth step

To a solution of compound 2c (61 mg, 110.01 micromole) in tetrahydrofuran (1 mL) and methanol (1 mL) was added an aqueous solution (0.5 mL) of sodium hydroxide (13.20 mg, 331.02 micromole), and the mixture was Stir at 28°C for 1.5 hours. Add 1 mol/L dilute hydrochloric acid to the reaction solution to adjust the pH to about 6, then extract with ethyl acetate (5 ml × 5), combine the organic phases and wash with water (5 ml × 3), collect the organic phases with anhydrous sulfuric acid Dry over sodium, filter, concentrate the filtrate, and purify the crude product by preparative chromatography (silica gel, dichloromethane:methanol=8:1) to obtain compound 3 .

MS-ESI calculated [M+H] ⁺ : 541.0, found 541.1.

¹ H NMR (400 MHz, DMSO- d ₆ ) δ = 14.31 - 14.25 (m, 1H), 12.10 (br d, J = 7.6 Hz, 1H), 7.66 (dd, J = 2.0, 5.6 Hz, 1H), 7.59 - 7.55 (m, 1H), 7.55 - 7.49 (m, 1H), 7.39 (d, J = 2.8 Hz, 1H), 6.98 - 6.94 (m, 1H), 5.20 - 5.15 (m, 1H), 5.11 - 5.07 (m, 1H), 3.83 (s, 3H), 2.75 (s, 3H).

Example 4

first step

To a solution of compound 4a (5-fluoro-2-methoxyphenol, 4.5 g, 31.66 mmol) in N,N-dimethylacetamide (30 mL) at 20 °C was added potassium hydroxide ( 3.55 g, 63.32 mmol), and bromoacetate aldehyde (9.36 g, 47.49 mmol) was added dropwise. The reaction system was warmed to 160°C and stirred at 160°C for 4 hours. Add water (10 ml) to the reaction system, use ethyl acetate (20 ml x 3), combine the organic phases with 10% aqueous sodium hydroxide solution (10 ml x 2). The collected organic phase was dried with anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain compound 4b .

¹ H NMR (400 MHz, CDCl ₃ ) δ = 6.82 - 6.76 (m, 1H), 6.74 - 6.68 (m, 1H), 6.64 - 6.56 (m, 1H), 4.92 - 4.80 (m, 1H), 4.05 - 4.00 (d, J = 20.0, 2H), 3.83 (s, 3H), 3.79 - 3.73 (m, 2H), 3.68 - 3.62 (m, 2H), 1.24 (m, 6H).

second step

To a solution of compound 4b (9.72 g, 37.63 mmol) in toluene (110 mL) was added polyphosphoric acid (972 mg, 37.63 mmol). The mixture was stirred at 110°C for 2.5 hours. Water (10 ml) was added to the reaction solution, ethyl acetate (20 ml×3) was used, and the combined organic phases were washed with brine (10 ml×3). The collected organic phase was dried with anhydrous sodium sulfate, filtered, and the filtrate was concentrated and separated by column chromatography (silica gel, petroleum ether: ethyl acetate = 1:0-400:1) to obtain compound 4c .

¹ H NMR (400 MHz, CDCl ₃ ) δ = 7.61 (d, J = 2.4 Hz, 1H), 6.88 - 6.86 (d, J = 2.0 Hz, 1H), 6.85 - 6.82 (d, J = 9.2 Hz, 1H ), 6.73 - 6.67 (m, 1H), 4.00 (s, 3H).

third step

To a solution of compound 4c (1.2 g, 7.22 mmol) in acetic acid (110 mL) was added palladium on carbon (120 mg, 10% purity) under nitrogen atmosphere. The reaction mixture was replaced with hydrogen and the mixture was stirred under hydrogen atmosphere (15 psi) at 25°C for 32 hours. The mixture was filtered through celite. Palladium on carbon (100 mg, 10% purity) was added to the filtrate, and the mixture was stirred under hydrogen atmosphere (15 psi) at 30°C for 16 hours. The mixture was filtered with celite, and the filtrate was concentrated to obtain compound 4d .

¹ H NMR (400 MHz, CDCl ₃ ) δ = 6.71 - 6.62 (m, 1H), 6.54 - 6.45 (m, 1H), 4.74 - 4.62 (m, 2H), 3.89 - 3.79 (s, 3H), 3.33 - 3.22 (m, 2H).

the fourth step

At -78 °C, boron tribromide (2.02 g, 8.06 mmol) was added to a solution of compound 4d (1.13 g, 6.72 mmol) in dichloromethane (10 ml), and the reaction system was warmed to 20 °C , and stirred at 20 °C for 2 hours. At 0°C, slowly add saturated ammonium chloride aqueous solution (10 ml) to the reaction solution to quench the reaction, and add water (10 ml) to the reaction system, wash with ethyl acetate (10 ml × 3), combine the organic The phase was washed with saline (10 ml × 3). The collected organic phase was dried over anhydrous sodium sulfate, filtered and concentrated to obtain compound 4e .

the fifth step

To a solution of compound 4e (1.94 g, 10.06 mmol) and 3-chloromethyl-1,2-difluoro-4-methoxybenzene (1.55 g, 10.06 mmol) in toluene (10 mL), An aqueous solution (2 mL) of tetrabutylammonium bromide (324.17 mg, 1.01 mmol) and sodium hydroxide (804.42 mg, 20.11 mmol) was added. The mixture was stirred at 50°C for 12 hours. 10 mL of water was added to the reaction system, extracted with ethyl acetate (20 mL×3), and the combined organic phases were washed with water (10 mL×3) and brine (10 mL×3). The collected organic phase was dried with anhydrous sodium sulfate, concentrated by filtration, and separated by column chromatography (silica gel, petroleum ether: ethyl acetate = 1:0-80:1) to obtain compound 4f .

¹ H NMR (400 MHz, CDCl ₃ ) δ = 7.14 - 7.09 (m, 1H), 6.81 - 6.78 (m, 1H), 6.62 - 6.61 (m, 1H), 6.50 - 6.46 (m, 1H), 5.15 - 5.14 (d, J = 16.0 Hz, 2H), 4.69 – 4.64 (m, 2H), 3.83 (s, 3H), 3.29 – 3.24 (m, 2H).

step six

To a solution of compound 4f (1.31 g, 4.22 mmol) in acetic acid (1 mL) at 80 °C, nitric acid (532.08 mg, 60% purity, 5.07 mmol) was added dropwise, and the mixture was cooled at 80 °C Stir for 2 hours. The reaction solution was poured into 50 ml of water, solids were precipitated, concentrated by filtration, and then slurried with 10 ml of mixed solvent (petroleum ether: ethyl acetate = 10:1) at 20°C for 30 minutes to obtain compound 4g .

¹ H NMR (400 MHz, CDCl ₃ ) δ = 7.85 - 7.79 (d, J = 6.4 Hz, 1H), 7.19 - 7.10 (m, 1H), 6.66 - 6.60 (m, 1H), 5.25 (s, 2H) , 4.89 - 4.77 (m, 2H), 3.88 (s, 3H), 3.41 - 3.32 (m, 2H).

step seven

Under a nitrogen atmosphere, palladium carbon (120 mg, 10% purity) was added to a solution of compound 4g (1.2 g, 3.38 mmol) in ethyl acetate (5 ml), and after hydrogen replacement, the mixture was heated under a hydrogen atmosphere ( 15 psi) and 25°C for 16 hours. The reaction solution was filtered with diatomaceous earth, and the filtrate was concentrated to obtain compound 4h .

¹ H NMR (400 MHz, CDCl ₃ ) δ = 7.14 - 7.05 (m, 1H), 6.83 - 6.78 (m, 1H), 6.60 - 6.55 (m, 1H), 5.13 - 5.12 (d, J = 1.6 Hz, 2H), 4.67 - 4.58 (m, 2H), 3.82 (s, 3H), 3.29 - 3.19 (m, 2H); MS-ESI calculated value [M+H] ⁺ : 326.1, found value 326.1.

eighth step

To a solution of compound 4h (100 mg, 307.43 micromol) in THF (3 mL), compound 1f (103 mg, 307.43 micromol) and triethylamine (31.11 mg, 307.43 micromol) were added, and the mixture was Stir at 70°C for 10 hours. The reaction solution was poured into 10 ml of water, extracted with ethyl acetate (30 ml × 5), the combined organic phases were washed with water (10 ml × 3), dried over anhydrous sodium sulfate, filtered, concentrated, and the crude product was purified by preparative chromatography ( Silica gel, petroleum ether: ethyl acetate = 3:1) to obtain compound 4i .

¹ H NMR (400 MHz, CDCl ₃ ) δ = 8.82 (s, 1H), 7.96 (s, 1H), 7.32 - 7.28 (m, 1H), 7.14 - 7.07 (m, 1H), 6.60 - 6.54 (m, 1H), 6.27 (br s, 1H), 5.19 - 5.18 (d, J = 1.6 Hz, 2H), 4.74 - 4.64 (m, 2H), 3.91 (s, 3H), 3.86 (s, 3H), 3.82 ( s, 3H), 3.33 - 3.25 (m, 2H); MS-ESI calculated value [M+H] ⁺ : 567.1, found value 567.1.

Ninth step

To a solution of compound 4i (102 mg, 180.05 micromol) in tetrahydrofuran (2 ml) and methanol (1 ml) was added an aqueous solution (1 ml) of lithium hydroxide monohydrate (22.66 mg, 540.16 micromol), the The mixture was stirred at 26°C for 1 hour. Add 1 ml of water to the reaction solution, adjust the pH to about 5 with 1 mol/L hydrochloric acid, then extract with ethyl acetate (10 ml × 3), combine the organic phases and wash with brine (10 ml × 2) , the collected organic phase was dried with anhydrous sodium sulfate, filtered, concentrated, and the crude product was prepared and separated by high performance liquid chromatography (column: Phenomenex luna C18 150*25 mm* 10 μm; mobile phase: [water (0.05% hydrochloric acid)-acetonitrile] ; Acetonitrile%: 42%-72%) to obtain compound 4 .

¹ H NMR (400 MHz, DMSO-d ₆ ) δ = 11.99 (br s, 1H), 7.54 - 7.41 (m, 1H), 7.36 (s, 1H), 7.13 - 7.12 (d, J=6.0 Hz, 1H ), 6.94 - 6.84 (m, 1H), 5.00 (s, 2H), 4.73 - 4.64 (m, 2H), 3.81 (s, 3H), 3.31 - 3.30 (m, 2H); calculated by MS-ESI [M +H] ⁺ : 521.1, found value 521.0.

Example 5

first step

To a solution of compound 5a (5-fluoro-2-methoxyphenol, 3 g, 21.11 mmol) in N,N-dimethylformamide (20 mL) was added cesium carbonate (13.75 g, 42.22 mmol ears), and the reaction system was stirred at 70°C for 1 hour. Then 1,2-dibromoethane (39.65 g, 211.08 mmol) was added to the reaction solution, and the reaction system was stirred at 70° C. for 18 hours. The reaction system was poured into water (30 mL), extracted with ethyl acetate (50 mL×5), and the combined organic phases were washed with water (10 mL×5). The collected organic phase was dried with anhydrous sodium sulfate, concentrated by filtration, and the crude product was purified by column chromatography (silica gel, petroleum ether: ethyl acetate = 1:0-200:1) to obtain compound 5b .

¹ H NMR (400 MHz, CDCl ₃ ) δ = 6.86 - 6.80 (m, 1H), 6.72 - 6.61 (m, 2H), 4.32 (t, J = 6.6 Hz, 2H), 3.86 (s, 3H), 3.67 (t, J = 6.6 Hz, 2H).

second step

To a solution of compound 5b (2 g, 8.03 mmol) in dimethyloxide (20 mL) was added potassium tert-butoxide (1.80 g, 16.06 mmol). The mixture was stirred at 25°C for 0.5 hours. The reaction system was poured into 20 ml of water, extracted with dichloromethane (50 ml x 5), and the combined organic phases were washed with water (10 ml x 5) and once with saturated brine. The collected organic phase was dried over anhydrous sodium sulfate, filtered and concentrated to obtain compound 5c .

third step

To a solution of compound 5c (900 mg, 5.35 mmol) in dichloromethane (10 mL) was added diiodomethane (151.60 g, 192.67 mmol) at 0°C. Then, diethylzinc (96.33 ml, 1 M) was added dropwise to the reaction system at 0°C. The mixture was warmed slowly to 25°C and stirred at 25°C for 12 hours. At 0°C, add 3 milliliters of saturated ammonium chloride aqueous solution and 20 milliliters of ethyl acetate to the reaction system, then pour the mixture into 10 milliliters of water, separate the layers, and then wash the aqueous phase with ethyl acetate (10 milliliters). ×5) extraction, the organic phases were combined and washed with water (10 ml ×5) and once with brine. The collected organic phase was dried with anhydrous sodium sulfate, concentrated by filtration, and the crude product was purified by column chromatography (silica gel, petroleum ether: ethyl acetate = 1:0-100:1) to obtain compound 5d .

¹ H NMR (400 MHz, CDCl ₃ ) δ = 7.03 (dd, J = 3.2, 10.0 Hz, 1H), 6.78 (dd, J = 5.2, 8.8 Hz, 1H), 6.62 (m, 1H), 3.83 (s , 3H), 3.76 - 3.70 (m, 1H), 0.89 - 0.85 (m, 2H), 0.85 - 0.78 (m, 2H).

the fourth step

To a solution of compound 5d (856 mg, 4.70 mmol) in dichloromethane (3 mL) was added boron tribromide (1.29 g, 5.17 mmol) in dichloromethane (0.5 mL) at -70 °C. solution, the reaction system was warmed to 0°C and stirred at 0°C for 1 hour. Methanol (2 ml) and water (10 ml) were slowly added dropwise to the reaction solution, extracted with dichloromethane (30 ml × 3), the combined organic phases were washed with water (10 ml × 3), washed once with brine, and collected The organic phase was dried over anhydrous sodium sulfate, filtered and concentrated to obtain compound 5e .

¹ H NMR (400 MHz, CDCl ₃ ) δ = 6.96 (dd, J = 3.2, 10 Hz, 1H), 6.83 (dd, J = 5.2, 8.8 Hz, 1H), 6.59 (m, 1H), 5.28 (br s, 1H), 3.87 - 3.72 (m, 1H), 0.89 - 0.81 (m, 4H).

the fifth step

To a solution of compound 5e (200 mg, 1.19 mmol) and 2-chloromethyl-3,4-difluoro-1-methoxybenzene (343.57 mg, 1.78 mmol) in toluene (5 mL), Tetrabutylammonium bromide (38.34 mg, 118.93 micromoles) was added. A solution of sodium hydroxide (95.14 mg, 2.38 mmol) in water (1 ml) was then added to the reaction. The mixture was stirred at 50°C for 12 hours. The reaction system was poured into 10 mL of water, extracted with ethyl acetate (20 mL×3), and the combined organic phases were washed with water (10 mL×3) and brine (10 mL×3). The collected organic phase was dried over anhydrous sodium sulfate, concentrated by filtration, and the crude product was purified by column chromatography (silica gel, petroleum ether: ethyl acetate = 1:0-80:1) to obtain compound 5f .

¹ H NMR (400 MHz, CDCl ₃ ) δ = 7.10 (q, J = 9.2 Hz, 1H), 6.97 (dd, J = 3.2, 10.4 Hz, 1H), 6.92 (dd, J = 5.6, 8.8 Hz, 1H ), 6.61 - 6.57 (m, 1H), 6.56 (td, J = 1.6, 3.0 Hz, 1H), 5.09 (d, J = 2.0 Hz, 2H), 3.80 (s, 3H), 3.75 - 3.68 (m, 1H), 0.81 - 0.76 (m, 4H).

step six

To a solution of compound 5f (281 mg, 866.50 μmol) in acetic acid (10 mL) was added nitric acid (136.50 mg, 97.50 μl, 60% purity) at 80 °C, and the mixture was stirred at 80 °C for 2 h . The reaction solution was poured into 10 ml of water, and a solid precipitated out. Filter and dry the filter cake to obtain compound 5g .

¹ H NMR (400 MHz, CDCl ₃ ) δ = 7.86 (d, J = 7.6 Hz, 1H), 7.18 - 7.07 (m, 2H), 6.65 - 6.59 (m, 1H), 5.22 (d, J = 1.6 Hz , 2H), 3.88 (s, 3H), 3.84 - 3.77 (m, 1H), 0.95 - 0.88 (m, 2H), 0.88 (s, 2H).

step seven

To a solution of compound 5g (207 mg, 560.53 micromolar) in ethanol (5 mL) and water (0.5 mL), iron powder (156.53 mg, 2.80 micromolar) and ammonium chloride (17.99 mg, 336.32 micromolar Ear). The mixture was stirred at 85°C for 1 hour. The reaction solution was filtered with diatomaceous earth, the filter cake was washed with dichloromethane (10 ml × 5), then water (10 ml) was added to the filtrate, extracted with dichloromethane (10 ml × 3), and the combined The organic phase was washed with water (10 ml × 2), dried over anhydrous sodium sulfate, concentrated by filtration, and the crude product was purified by column chromatography (silica gel, petroleum ether: ethyl acetate = 1:0-10:1) to obtain compound 5h .

¹ H NMR (400 MHz, CDCl ₃ ) δ = 7.10 (q, J = 9.6 Hz, 1H), 6.92 (d, J = 12.0 Hz, 1H), 6.62 - 6.56 (m, 1H), 6.52 (d, J = 8.8 Hz, 1H), 5.06 (s, 2H), 3.82 (s, 3H), 3.69 (tt, J = 3.2, 6.0 Hz, 1H), 3.42 (br s, 2H), 0.80 - 0.72 (m, 2H ), 0.72 - 0.64 (m, 2H); MS-ESI calculated value [M+H] ⁺ : 340.1, found value 340.1.

eighth step

To a solution of compound 5h (220 mg, 583.54 μM) in THF (5 mL), compound 1f (195.68 mg, 583.54 μM) and triethylamine (59.05 mg, 583.54 μM) were added. The mixture was stirred at 70°C for 10 hours. The reaction solution was poured into 30 ml of water, extracted with ethyl acetate (50 ml × 5), the combined organic phase was washed with water (20 ml × 5), saturated brine (20 ml), dried over anhydrous sodium sulfate, filtered , concentrated, and the crude product was purified by column chromatography (silica gel, petroleum ether: ethyl acetate = 100:1-5:1) to obtain compound 5i .

¹ H NMR (400 MHz, CDCl ₃ ) δ = 8.79 (s, 1H), 7.96 (s, 1H), 7.58 (d, J = 7.6 Hz, 1H), 7.14 - 7.06 (m, 1H), 7.04 (d , J = 7.6 Hz, 1H), 6.60 - 6.54 (m, 1H), 6.43 (s, 1H), 5.15 (d, J = 1.6 Hz, 2H), 3.91 (s, 3H), 3.86 (s, 3H) , 3.81 (s, 3H), 3.73 (m, 1H), 0.91 - 0.85 (m, 2H), 0.78 - 0.74 (m, 2H); MS-ESI calculated value [M+H] ⁺ : 581.1, measured value 581.1 .

Ninth step

To a solution of compound 5i (324 mg, 533.16 micromol) in THF (1 mL) and methanol (1 mL) was added an aqueous solution (0.5 mL) of lithium hydroxide monohydrate (134.23 mg, 3.20 mmol), the The mixture was stirred at 28°C for 2 hours. Add 1 mol/L dilute hydrochloric acid to the reaction solution to adjust the pH to about 6, and a solid precipitates, which is filtered and dried to obtain compound 5 .

¹ H NMR (400 MHz, DMSO- d ₆ ) δ = 14.52 (br s, 1H), 12.01 (br s, 1H), 7.50 - 7.41 (m, 1H), 7.39 (s, 1H), 7.33 (d, J = 11.2 Hz, 1H), 7.26 (m, 1H), 6.93 - 6.86 (m, 1H), 4.95 (s, 2H), 3.91 (m, 1H), 3.78 (s, 3H), 0.87 - 0.77 (m , 2H), 0.73 - 0.65 (m, 2H); MS-ESI calculated value [M+H] ⁺ : 535.1, found value 535.0.

Example 6

first step

Compound 6a (2-bromo-4-fluoro-5-nitrophenol, 2 g, 8.47 mmol) was dissolved in 15 ml of N-methylpyrrolidone, cooled to 0 °C, and potassium tert-butoxide (1.14 g, 10.17 mmol), then stirred at 0°C for 30 minutes, and the compound 2-chloromethyl-3,4-difluoro-1-methoxybenzene (2.77 g, 14.41 mmol) was added dropwise into the reaction solution and stirred at 25°C for 3 hours. The reaction solution was poured into 100 ml of water, and extracted three times with ethyl acetate, 30 ml each time, the combined organic phase was concentrated, and the crude product was subjected to column chromatography (silica gel, 100-200 mesh, petroleum ether: ethyl acetate=1/0 -10/1) to obtain compound 6b .

second step

Compound 6b (300 mg, 765.06 micromol), cyclopropylboronic acid (131.43 mg, 1.53 mmol), palladium acetate (17.18 mg, 76.51 micromol), tri(cyclohexyl)phosphine (42.91 mg, 153.01 micromoles), potassium phosphate (487.19 mg, 2.30 mmoles) was dissolved in 10 ml of toluene and 1 ml of water, and replaced with nitrogen three times, then heated to 100°C and stirred at 100°C for 12 hours. The reaction solution was filtered and concentrated, and the crude product was purified by column chromatography (silica gel, 100-200 mesh, petroleum ether: ethyl acetate = 1/0-10/1) to obtain compound 6c .

third step

Compound 6c (200 mg, 566.10 micromol) was dissolved in 6 mL of ethanol and 0.6 mL of water, iron powder (158.08 mg, 2.83 mmol) and ammonium chloride (18.17 mg, 339.66 micromol) were added, Then the reaction solution was heated to 85°C and stirred at 85°C for 2 hours. The reaction solution was filtered and concentrated, and the crude product was dissolved in 20 ml of ethyl acetate, washed once with 20 ml of water and once with 10 ml of saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated to obtain compound 6d .

the fourth step

Compound 6d (140 mg, 433.03 micromoles) was dissolved in 4 ml of THF, compound 1f (145.21 mg, 433.02 micromoles) and triethylamine (43.82 mg, 433.02 micromoles) were added, and heated to 70 °C, stirred at 70 °C for 12 hours. The reaction solution was concentrated, and the crude product was purified by column chromatography (silica gel, 100-200 mesh, petroleum ether: ethyl acetate = 1/0-10/1) to obtain compound 6e .

MS-ESI calculated [M+Na] ⁺ : 587.1, found 587.1.

the fifth step

Compound 6e (120 mg, 212.57 micromole) was dissolved in 6 mL of tetrahydrofuran and 1 mL of methanol, then lithium hydroxide monohydrate (53.52 mg, 1.28 mmol) was dissolved in 1 mL of water, and added dropwise into the reaction solution and stirred at 25°C for 1 hour. Use 1 mol/liter of dilute hydrochloric acid to adjust the reaction solution to pH 2-3, then dilute with 10 ml of water, extract 3 times with ethyl acetate, 10 ml each time, combine the organic phases, and add 10 ml of saturated saline Washed once, dried with anhydrous sodium sulfate, filtered, and concentrated to obtain compound 6 .

1H NMR (400 MHz, DMSO- d ₆ ) δ = 7.47 (q, J = 9.6 Hz, 1H), 7.40 - 7.38 (m, 1H), 7.39 (s, 1H), 7.31 (d, J = 6.4 Hz, 1H), 6.94 - 6.89 (m, 1H), 6.86 (d, J = 11.2 Hz, 1H), 5.03 (s, 2H), 3.82 (s, 3H), 2.11 - 2.03 (m, 1H), 0.97 - 0.87 (m, 2H), 0.73 - 0.68 (m, 2H); MS-ESI calculated value [M+H] ⁺ : 519.1, found value = 519.0.

Example 7

first step

To N,N-dimethylformamide of compound 7a (methylpyruvate, 21 g, 205.70 mmol) and ethyl cyanoacetate (23.27 g, 205.70 mmol) at 0–5 °C Sulfur powder (7.92 g, 246.84 mmol) was added to the solution (200 ml), triethylamine (35.39 g, 349.70 mmol) was added dropwise, and the mixture was stirred at 50°C for 2 hours. Add water (500 ml) to the reaction solution, extract twice with ethyl acetate, 100 ml each time, wash the combined organic phase twice with water, 200 ml each time, wash with saturated brine (200 ml), dry over anhydrous sodium sulfate, and filter , the filtrate was concentrated to dryness under reduced pressure, ethyl acetate (5 ml) and petroleum ether (50 ml) were added, stirred at 30°C for 30 minutes, filtered, and the filter cake was dried under reduced pressure to obtain compound 7b .

second step

To a solution of compound 7b (28 g, 122.14 mmol) in acetonitrile (300 mL) was added cuprous bromide (26.28 g, 183.20 mmol), tert-butyl nitrite (15.11 g , 146.56 mmol), and the reaction mixture was stirred at 25°C for 12 hours. Water (100 ml) was added to the reaction solution, extracted twice with ethyl acetate, 100 ml each time, the combined organic phase was washed with saturated brine (100 ml), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to dryness. Purified by column chromatography (petroleum ether: ethyl acetate = 1:0 to 20:1) to obtain compound 7c .

third step

At -20°C, in a solution of compound 7c (4.5 g, 15.35 mmol) in dichloromethane (15 ml), boron tribromide (4.62 g, 18.42 mmol) was slowly added dropwise, and the reaction solution was heated at 25 °C under nitrogen protection and stirred for 12 hours. The reaction solution was filtered, and the filter cake was washed twice with dichloromethane, 10 ml each time, and the filter cake was discarded. The filtrate was diluted with water (10 ml), adjusted to pH 10-11 with 1.0 mol/L sodium hydroxide, and the dichloromethane phase was discarded. Dichloromethane (10 ml) was added to the aqueous phase, and the pH was adjusted to 3-4 with 1.0 mol/L hydrochloric acid. The dichloromethane phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to dryness to obtain compound 7 d .

the fourth step

To compound 7d (300 mg, 1.07 mmol) in tert-butanol (2 mL), diphenylphosphoryl azide (354.96 mg, 1.29 mmol), triethylamine (163.15 mg, 1.61 mmol ). The reaction was stirred at 85°C for 2 hours under nitrogen protection. Add water (10 ml) to the reaction solution, extract twice with ethyl acetate, 10 ml each time, combine the organic layers, wash with saturated aqueous sodium chloride solution (10 ml), dry over anhydrous sodium sulfate, filter, and concentrate the filtrate to dryness under reduced pressure , purified by thin-layer chromatography on silica gel plate (petroleum ether: ethyl acetate = 20:1) to obtain compound 7e .

LC-MS: m/z = 249.9 [M+H-100] ⁺ .

the fifth step

To a solution of compound 7e (110 mg, 314.08 micromolar) and diethoxymethylphosphine (85.51 mg, 628.16 micromolar) in N,N-dimethylformamide (3 mL), potassium phosphate ( 200.01 mg, 942.24 micromoles), diphenylphosphine-9,9-dimethyloxaxanthene (36.35 mg, 62.82 micromoles), palladium acetate (7.05 mg, 31.41 micromoles). The reaction solution was stirred at 100° C. for 12 hours under nitrogen protection. Add water (20 ml) to the reaction solution, extract twice with ethyl acetate, 10 ml each time, combine the organic phases, wash with saturated brine (20 ml), dry over anhydrous sodium sulfate, filter, and concentrate the filtrate to dryness. Preparative chromatographic plate (petroleum ether: ethyl acetate = 1:2) was purified to obtain compound 7f .

¹ H NMR (400MHz, CDCl ₃ ) δ = 9.13 (br s, 1H), 8.04 (br s, 1H), 4.46 (dq, J = 2.0, 7.2 Hz, 2H), 4.21 - 4.11 (m, 1H), 4.00 - 3.93 (m, 1H), 1.88 (d, J = 16.0 Hz, 3H), 1.52 (s, 9H), 1.47 (t, J = 7.2 Hz, 3H), 1.32 (t, J = 7.2 Hz, 3H ).

step six

To a solution of compound 7f (20 mg, 53.00 micromol) in dichloromethane (0.5 ml), trifluoroacetic acid (770.00 mg, 6.75 mmol) was added, and the reaction solution was stirred at 25°C for 1 hour. The reaction solution was concentrated to dryness under reduced pressure to obtain compound 7g (trifluoroacetate).

LC-MS: m/z = 278.1 [M+H] ⁺ .

step seven

Add potassium carbonate (7.95 mg, 57.50 micromol) to a solution of compound 7g (15 mg, 38.33 micromol, trifluoroacetate salt) in tetrahydrofuran (0.5 ml) and water (0.5 ml), at 5-10°C Phenyl chloroformate (12.00 mg, 76.67 micromoles) was added dropwise and the reaction was stirred at 5-10°C for 1 hour. After the reaction was monitored by liquid chromatography-mass chromatography, the reaction liquid was added with water (10 ml), extracted twice with ethyl acetate, 10 ml each time, the combined organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was reduced to Concentrate to dryness under pressure to obtain 7h .

LC-MS: m/z = 398.1 [M+H] ⁺ .

eighth step

To a solution of compound 7k (3,4-difluoroanisole, 10 g, 69.39 mmol) in THF (100 mL) at -70--65 °C, 2.0 mol/L of diisopropylamine was added lithium (41.63 ml), stirred at -70-65°C for 0.5 hour, and then a solution of N,N-dimethylformamide (6.09 g, 83.26 mmol) in tetrahydrofuran (5 ml) was added dropwise. The mixture was stirred at -70-65°C for 1 hour. At -70--65°C, add a mixture of acetic acid (10 ml) and water (100 ml), extract twice with ethyl acetate, 100 ml each time, wash the combined organic phase with 100 ml saturated brine, anhydrous sulfuric acid Dry over sodium, filter, and concentrate the filtrate to dryness under reduced pressure to obtain compound 7l .

Ninth step

To a solution of compound 7l (12 g, 69.72 mmol) in toluene (72 mL) at 10-15 °C was added sodium borohydride (5.27 g, 139.43 mmol) in 0.1 mol/L of NaOH (15 mL) solution. The reaction solution was stirred at 15°C for 4 hours. Add 100 ml of water, extract twice with ethyl acetate, 100 ml each time, wash the combined organic phase with 100 ml of saturated brine, dry over anhydrous sodium sulfate, filter, and concentrate the filtrate under reduced pressure to dryness to obtain compound 7m .

1H NMR (400MHz, CDCl ₃ ) δ = 7.07 (q, J = 9.1 Hz, 1H), 6.63 - 6.54 (m, 1H), 4.77 (dd, J = 1.5, 6.4 Hz, 2H), 3.88 (s, 3H ).

tenth step

To compound 7m (12 g, 68.91 mmol) in toluene (120 ml) was added argonium chloride (12.30 g, 103.36 mmol), and the reaction was stirred at 25°C for 2 hours under nitrogen protection. The reaction solution was concentrated to dryness, 50 ml of water was added, the pH was adjusted to 7-8 with saturated sodium bicarbonate, extracted twice with ethyl acetate, 50 ml each time, the organic layers were combined, washed once with 50 ml of saturated sodium chloride aqueous solution, anhydrous Dry over sodium sulfate, filter, and concentrate the filtrate to dryness under reduced pressure to obtain compound 7n .

Eleventh step

To compound 7n (1 g, 5.19 mmol) and 4-fluoro-2-methoxyphenol (737.98 mg, 5.19 mmol) in toluene (40 ml), add tetrabutylammonium bromide (167.38 mg , 519.24 μmol), sodium hydroxide (415.36 mg, 10.38 mmol) in water (2 ml) solution, and the reaction solution was stirred at 50°C for 12 hours. The reaction solution was added with 10 ml of water, extracted twice with 10 ml of ethyl acetate each time, the organic phases were combined, washed with 10 ml of saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to dryness to obtain compound 7p .

Twelfth step

To a solution of compound 7p (1.7 g, 5.70 mmol) in acetic acid (12 mL), nitric acid (990 mg, 9.43 mmol, 60% purity) was added dropwise, and the reaction was stirred at 60 °C for 0.5 h. After adding 50 ml of water, a solid precipitated out, filtered, and the filter cake was dried under reduced pressure to obtain 7q .

LC-MS: m/z = 344.1 [M+H] ⁺ .

Step Thirteen

To a solution of compound 7q (1.8 g, 5.24 mmol) in ethyl acetate (150 mL), wet palladium on carbon (180 mg, 10% purity, 50% water) was added, and the reaction was stirred at 20 °C under hydrogen at 15 psi for 12 hours . The reaction liquid was filtered, and the filtrate was concentrated to dryness under reduced pressure, ethyl acetate (5 ml) and petroleum ether (50 ml) were added, slurried at 20°C for 30 minutes, filtered, and dried under reduced pressure to obtain 7j .

LC-MS: m/z = 314.1 [M+H]+.

step fourteen

To a solution of compound 7h (18 mg, 45.30 micromol) and compound 7j (14.19 mg, 45.30 micromol) in THF (2 mL) was added triethylamine (4.58 mg, 45.30 micromol) and reacted at 33 Stir at °C for 120 hours. The reaction solution was concentrated to dryness under reduced pressure, and purified by preparative chromatography (petroleum ether: ethyl acetate = 1:2) to obtain 7i .

LC-MS: m/z = 617.1 [M+H]+.

Step fifteen

To a solution of compound 7i (7 mg, 11.35 micromoles) in tetrahydrofuran (0.5 ml), methanol (0.5 ml) and water (0.5 ml) was added monohydrate and lithium hydroxide (2.38 mg, 56.77 micromoles), The reaction was stirred at 20°C for 2 hours. The pH of the reaction solution was adjusted to 3-4 with 1 mole per liter of hydrochloric acid, the solid was precipitated, filtered, washed twice with 2 ml of water each time, and the filter cake was concentrated to dryness to obtain compound 7 .

LC-MS: m/z = 543.1 [M+H] ⁺ ; ¹ H NMR (400MHz, CD ₃ OD) δ = 7.27 - 7.19 (m, 1H), 7.14 (s, 1H), 7.05 (br d, J = 7.2 Hz, 1H), 6.95 (d, J = 11.2 Hz, 1H), 6.78 (br d, J = 9.4 Hz, 1H), 5.10 (s, 2H), 3.85 (s, 3H), 3.80 (s, 3H), 1.75 (br d, J = 15.2 Hz, 3H).

Example 8

first step

To a solution of compound 8a (5,6-difluorosalicylaldehyde, 3 g, 18.98 mmol) in DMF (30 mL) was added cesium carbonate (12.36 g, 37.95 mmol), and the reaction system was stirred at 70°C for 1 hour. Then 1,2-dibromoethane (17.82 g, 94.88 mmol) was added to the reaction solution, and the reaction system was stirred at 70° C. for 18 hours. The reaction system was poured into water (30 mL), extracted with ethyl acetate (50 mL×5), and the combined organic phases were washed with water (20 mL×5) and brine. The collected organic phase was dried with anhydrous sodium sulfate, filtered, and the filtrate was concentrated and purified by column chromatography (silica gel, petroleum ether/ethyl acetate=100/1-20/1) to obtain compound 8b .

second step

To a solution of compound 8b (1.68 g, 6.34 mmol) in tetrahydrofuran (30 mL) and methanol (6 mL) was added sodium borohydride (287.74 mg, 7.61 mmol) at 0 °C. The mixture was stirred at 25°C for 0.5 hours. To the reaction liquid were added 10 mL of saturated aqueous ammonium chloride and 20 mL of ethyl acetate at 0°C, and the mixture was stirred at 25°C for 0.5 hr. Then the mixture was poured into 20 mL of water, extracted with ethyl acetate (50 mL×5), and the combined organic phases were washed with water (20 mL×5) and brine. The collected organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to obtain compound 8c .

¹ H NMR (400 MHz, CDCl ₃ ) δ = 7.06 (q, J = 9.0 Hz, 1H), 6.64 - 6.51 (m, 1H), 4.80 (br d, J = 5.0 Hz, 2H), 4.39 - 4.32 ( m, 2H), 3.74 - 3.69 (m, 2H), 2.60 (br t, J = 6.8 Hz, 1H).

third step

To a solution of compound 8c (1.83 g, 16.85 mmol) in dichloromethane (5 mL) at 0 °C, 2,6-lutidine (1.10 g, 1.5 eq ) and triisopropylsilane were added dropwise Trifluoromethylsulfate (trifluoromethylsulfate) ester (2.52 g, 1.2 eq). The mixture was warmed to 25°C and stirred at 25°C for 12 hours. To the reaction solution were added 1 mL of saturated aqueous ammonium chloride solution and 10 mL of ethyl acetate at 0°C, and the mixture was stirred at 25°C for 0.5 hr. Then the mixture was poured into 10 mL of water, extracted with ethyl acetate (50 mL×5), and the combined organic phases were washed with water (10 mL×5) and brine. The collected organic phase was dried with anhydrous sodium sulfate, filtered, and the filtrate was concentrated and purified by column chromatography (silica gel, petroleum ether/ethyl acetate=1/0-50/1) to obtain compound 8d .

¹ H NMR (400 MHz, CDCl ₃ ) δ = 7.04 (q, J = 9.2 Hz, 1H), 6.62 - 6.54 (m, 1H), 4.87 (d, J = 2.0 Hz, 2H), 4.29 (t, J = 6.4 Hz, 2H), 3.63 (t, J = 6.4 Hz, 2H), 1.20 - 1.13 (m, 3H), 1.12 - 1.08 (m, 18H).

the fourth step

To a solution of compound 8d (2.97 g, 7.01 mmol) in tetrahydrofuran (3 mL) was added potassium tert-butoxide (1.57 g, 14.03 mmol) at 0°C. The mixture was stirred at 60°C for 24 hours. The reaction system was poured into 10 mL of water, extracted with ethyl acetate (10 mL×5), and the combined organic phases were washed with water (10 mL×5) and brine. The collected organic phase was dried with anhydrous sodium sulfate, filtered, and the filtrate was concentrated and purified by column chromatography (silica gel, petroleum ether/ethyl acetate=1/0) to obtain compound 8e .

the fifth step

To a solution of compound 8e (1.8 g, 5.26 mmol) in dichloromethane (10 mL) was added chloroiodomethane (33.37 g, 189.20 mmol) at 0 °C. Then, diethylzinc (94.60 mL, 1M, 94.60 mmol) was added dropwise to the reaction system at 0°C. The mixture was warmed slowly to 25°C and stirred at 25°C for 12 hours. At 0°C, 1 mL of saturated ammonium chloride aqueous solution and 10 mL of ethyl acetate were added to the reaction system, and then the mixture was poured into 10 mL of water, and the aqueous phase was extracted with ethyl acetate (10 mL×5). The combined organic phases were washed with water (10 mL×5) and brine. The collected organic phase was dried with anhydrous sodium sulfate, filtered, and the filtrate was concentrated and purified by column chromatography (silica gel, petroleum ether/ethyl acetate=1/0) to obtain compound 8f .

step six

To a solution of compound 8f (669 mg, 1.88 mmol) in tetrahydrofuran (3 mL) was added TBAF (2.25 mL, 1 M, 2.25 mmol), and the mixture was stirred at 27°C for 1.3 hr. To the reaction solution, 10 mL of saturated aqueous sodium bicarbonate solution was added dropwise to quench the reaction. Then the reaction solution was poured into 10 mL of water, extracted with ethyl acetate (10 mL×5), the combined organic phase was washed with saturated aqueous sodium bicarbonate solution (10 mL×5), brine, dried over anhydrous sodium sulfate, and filtered , the filtrate was concentrated and purified by column chromatography (silica gel, petroleum ether/ethyl acetate = 1/0-20/1) to obtain compound 8g .

¹ H NMR (400 MHz, CDCl ₃ ) δ = 7.11 - 7.03 (m, 1H), 6.99 - 6.93 (m, 1H), 4.75 - 4.70 (m, 2H), 3.77 (td, J = 2.8, 6.0 Hz, 1H), 0.85 - 0.79 (m, 4H).

step seven

To a solution of compound 8g (518 mg, 2.59 mmol) in toluene (3 mL) was added phosphine chloride (307.85 mg, 2.59 mmol) under nitrogen atmosphere, and the mixture was stirred at 28°C for 2 hours. Add 10 mL of water to the reaction system, extract with ethyl acetate (20 mL × 3), combine the organic phases with water (10 mL × 3), wash with brine (10 mL × 3), collect the organic phase and wash with anhydrous sodium sulfate Drying, filtration and concentration gave crude compound 8h .

¹ H NMR (400 MHz, CDCl ₃ ) δ = 7.14 - 7.08 (m, 1H), 6.99 - 6.94 (m, 1H), 4.63 (d, J = 1.6 Hz, 2H), 3.83 - 3.77 (m, 1H) , 0.83 (d, J = 4.4 Hz, 4H).

eighth step

To a solution of compound 8i (137.30 mg, 966.03 μmmol) and compound 8h (528 mg, crude) in toluene (5 mL) was added tetrabutylammonium bromide (38.93 mg, 120.75 μmmol). Then a solution of sodium hydroxide (96.60 mg, 2.42 mmol) in water (1 mL) was added to the reaction system. The mixture was stirred at 50°C for 12 hours. The reaction system was poured into 10 mL of water, extracted with ethyl acetate (10 mL×5), and the combined organic phases were washed with water (10 mL×5) and brine. The collected organic phase was dried with anhydrous sodium sulfate, filtered, and the filtrate was concentrated and purified by column chromatography (silica gel, petroleum ether/ethyl acetate=1/0-100/1) to obtain compound 8j .

¹ H NMR (400 MHz, CDCl ₃ ) δ = 7.11 (q, J = 9.2 Hz, 1H), 6.97 - 6.91 (m, 1H), 6.85 - 6.81 (m, 1H), 6.63 (dd, J = 2.8, 9.8 Hz, 2H), 5.08 (d, J = 1.8 Hz, 2H), 3.82 (s, 3H), 3.75 - 3.67 (m, 1H), 0.80 - 0.74 (m, 2H), 0.73 - 0.67 (m, 2H ).

Ninth step

To a solution of compound 8j (90 mg, 277.3 μmmol) in dichloromethane (1 mL) was added a solution of concentrated nitric acid (53.53 mg, 98% purity, 832.58 μmmol) in dichloromethane (0.5 mL) at 0 °C, The mixture was stirred at 0°C for 3 hours. At 0°C, concentrated nitric acid (71.38 mg, 98% purity, 1.11 mmol) was added again to the reaction system, and the mixture was stirred at 0°C for 0.5 hours. Add saturated aqueous sodium bicarbonate solution to the reaction solution to adjust the pH to 7, then add 10 mL of water, and extract with dichloromethane (10 mL×3). The combined organic phases were washed with water (10 mL×2) and saturated brine (10 mL×2). The collected organic phase was dried over anhydrous sodium sulfate, filtered and concentrated to obtain compound 8k .

¹ H NMR (400 MHz, CDCl ₃ ) δ = 7.79 (d, J = 7.4 Hz, 1H), 7.20 - 7.09 (m, 1H), 7.07 - 6.96 (m, 1H), 6.71 (d, J = 12.4 Hz , 1H), 5.23 - 5.17 (m, 2H), 3.93 (s, 3H), 3.86 - 3.72 (m, 1H), 0.91 - 0.70 (m, 5H).

tenth step

To a solution of compound 8k (90 mg, 243.71 μmmol) in ethanol (3 mL) and water (0.3 mL), iron powder (68.06 mg, 1.22 mmol) and ammonium chloride (7.82 mg, 146.23 μmmol) were added. The mixture was stirred at 85°C for 1 hour. The reaction solution was filtered with diatomaceous earth, and the filter cake was washed with dichloromethane (10 mL×5), then water (10 mL) was added to the filtrate, extracted with dichloromethane (10 mL×3), and the organic phases were combined , washed with water (10 mL×2), brine (10 mL×2), dried over anhydrous sodium sulfate, filtered and concentrated to obtain compound 8l .

LC-MS: m/z = 340.1 [M+H] ⁺ ; ¹ H NMR (400 MHz, CDCl ₃ ) δ = 7.11 (q, J = 9.3 Hz, 1H), 7.04 - 6.90 (m, 1H), 6.64 (d, J = 12.0 Hz, 1H), 6.49 (d, J = 8.9 Hz, 1H), 5.04 (d, J = 1.6 Hz, 2H), 3.75 (s, 3H), 3.74 - 3.70 (m, 1H) , 0.79 - 0.75 (m, 2H), 0.75 - 0.69 (m, 2H).

Eleventh step

To a solution of compound 8l (70 mg, 206.30 μmmol) in tetrahydrofuran (5 mL), were added compound 1f (69.18 mg, 206.30 μmmol) and triethylamine (20.88 mg, 206.30 μmmol). The mixture was stirred at 70°C for 10 hours. Pour the reaction solution into 30 mL of water, extract with ethyl acetate (50 mL×5), combine the organic phases with water (20 mL×5), wash with saturated brine, and dry with anhydrous sodium sulfate, filter, and concentrate the filtrate Compound 8m was prepared and separated by thin layer chromatography (silica gel, petroleum ether/ethyl acetate = 2/1).

LC-MS: m/z = 581.2 [M+H] ⁺ .

Twelfth step

To a solution of compound 8m (55 mg, 94.74 μmmol) in tetrahydrofuran (1 mL) and methanol (1 mL) was added an aqueous solution (0.5 mL) of lithium hydroxide monohydrate (23.85 mg, 568.45 μmmol), and the mixture was heated at 28°C Stir for 2 hours. Hydrochloric acid was added to the reaction solution to adjust the pH to about 6. Then it was extracted with ethyl acetate (5 mL×5), the combined organic phases were washed with water (5 mL×3), saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated and separated by thin layer chromatography (silica gel, Dichloromethane/methanol = 20/1) to give compound 8 .

LC-MS: m/z = 535.1 [M+H] ⁺ ; ¹ H NMR (400 MHz, DMSO- d ₆ ) δ = 14.53 (br s, 1H), 11.99 (br s, 1H), 7.50~7.45 ( m, 1H), 7.37 (s, 1H), 7.21 - 7.10 (m, 3H), 4.92 (s, 2H), 3.94 - 3.88 (m, 1H), 3.81 (s, 3H), 0.78 - 0.71 (m, 2H), 0.66 - 0.59 (m, 2H). biological test data

Experimental example 1 Test of the activity of the compounds of the present invention on human gonadotropin-releasing hormone receptors

Purpose of the experiment: To use FLIPR detection technology to detect the inhibitory activity of the test compound on the gonadotropin-releasing hormone receptor at the cellular level. The main experimental materials and sources: Fluo-4 Direct ^TM Kit——Invitrogen-F10471 Company 384-well polymer Lysine-coated cell plate - Greiner-781946 384-well compound plate - Greiner-781280 compound preparation ECHO (acoustic pipetting system) - Labcyte FLIPR (fluorescence imaging plate reader) - Molecular Devices Experimental program:

Take GnRH/HEK293 (human embryonic kidney cell 293) cultured cells in the logarithmic growth phase, wash with DPBS (Duchenne's phosphate-buffered saline) buffer, add an appropriate amount of 0.05% EDTA (ethylenediaminetetraacetic acid)-trypsin and place at 37°C Digest in a carbon dioxide incubator, take out the cells for 1-2 minutes and add medium to terminate the digestion. After repeated blowing and blowing to disperse the cells, the cells were collected by centrifugation, and seeded into a 384-well polylysine-coated cell plate at a density of 20 μL at 20,000 cells per well, and incubated overnight in a 37°C incubator with 5% CO ₂ .

On the second day, 20 μL of 2× Fluo-4 Direct ^TM buffer was added to each well, 5% CO ₂ , incubated in a 37°C incubator for 50 minutes, and the cells were placed at room temperature for 10 minutes. A 4-fold 10-point serial dilution of 0.2 mM Leuprolide acetate (leuprolide acetate) was made with ECHO, and 900 nL was transferred to the compound plate. Add 30 μL of FLIPR buffered saline solution to the compound plate and centrifuge at 1000 rpm for 1 min. Run the FLIPR instrument software, add 10 μL experimental buffer salt solution according to the set program, and read the fluorescent signal. Then add 10 μL agonist reference compound, read the fluorescence signal, calculate EC ₈₀ , and prepare agonist with 6×EC ₈₀ concentration.

2 mM test compound and appropriate concentration of reference compound were serially diluted 4 times and 10 points with ECHO, and transferred 900 nL to the compound plate. Add 30 μL of FLIPR buffered saline solution to the compound plate and centrifuge at 1000 rpm for 1 min. Run the FLIPR instrument software, add 10 μL of test and reference compounds to the cell plate according to the set program, and read the fluorescent signal. Then add 10 μL of agonist at 6×EC ₈₀ concentration to the cell plate, and read the fluorescent signal.

Calculate the IC ₅₀ of the compound on the calcium flow inhibition of the gonadotropin-releasing hormone receptor, that is, the drug concentration at which the Ca ²⁺ flow in cells stably expressing the GnRH receptor is inhibited by half, and the IC ₅₀ of the drug is calculated using GraphPad Prism 5.0 software . Experimental results: The inhibitory activity of the compounds of the present invention on human gonadotropin-releasing hormone receptors was determined by the above test method, and the measured IC ₅₀ is shown in Table 1.

Table 1 Inhibitory results of compounds of the present invention on human gonadotropin-releasing hormone receptor activity Compound number _IC50 (nM) Compound 1 2.3 Compound 2 40.1 Compound 3 38.9 Compound 4 14.8 Compound 5 3.4 Compound 6 13.9 Compound 7 15.2 Compound 8 7.8 Conclusion: the compound of the present invention has significant inhibitory effect on human gonadotropin-releasing hormone receptor.

Experimental example 2 Pharmacokinetic Evaluation

Experimental purpose: To study the pharmacokinetic properties of the compound of the present invention in mice Test plan:

Each test compound was mixed with DMAC and vortexed for 2 minutes, and then the DMAC solution of the test compound was mixed and vortexed for 2 minutes to prepare a clear solution of 10 mg/mL. Take 0.0600 mL of 10 mg/mL solution and add 0.300 mL Solutol, vortex for 2 minutes, then add 2.400 mL of normal saline, vortex for 2 minutes to obtain a 0.2 mg/mL clear solution, which is used for PO group administration. Take 0.500 mL of PO group administration solution, vortex for 2 minutes, add 0.0500 mL of DMAC, mix and vortex for 2 minutes, then add 0.0500 mL of solutol, vortex for 2 minutes, finally add 0.400 mL of normal saline, vortex A clear solution of 0.1 mg/mL was obtained in two minutes, and the administration solution of the injection group (IV) was obtained after filtration through a microporous membrane.

Four male CD-1 mice were divided into 2 groups. Animals in group 1 were administered intravenously once, the dose was 0.5 mg/kg, the vehicle was 10% DMAC/10% Solutol/80% normal saline, and the administration volume was 5 mL/kg. Animals in group 2 were given a single oral gavage of 2 mg/kg of the test compound, the oral vehicle was 10% DMAC/10% Solutol/80% normal saline, and the oral volume was 10 mL/kg. Whole blood was collected at 0.033 (iv only), 0.083, 0.25, 0.5, 1, 2, 4, and 12 hours post-dose. Whole blood was centrifuged at 3200 g, 2-8°C for 10 min to obtain plasma, and the concentration of the test compound in plasma was determined by LC/MS/MS method, and the pharmacokinetic parameters were calculated by Phoenix WinNonlin software. Experimental results:

The test results are shown in Table 2. The meaning of each parameter: IV: intravenous injection; PO: oral administration; C ₀ : initial plasma concentration; C _max : maximum drug concentration in systemic circulation; T _max : time required to reach C _max ; T _1/2 : half-life; V _dss : apparent volume of distribution; Cl: clearance rate; AUC _0-last : area under the drug-time curve.

Table 2 Pharmacokinetic (PK) test results of compound 1 in plasma parameter IV (0.5 mg/kg) PO (2mg/kg) C ₀ (nmol/L) 17017 -- C _max (nmol/L) -- 22250 T _max (h) -- 0.750 T _1/2 (h) 14.9 16.9 V _dss (L/kg) 0.185 -- Cl(mL/min/kg) 0.162 -- AUC _0-last (h*nmol/L) 89645 251921 bioavailability(%) -- 77.0 "--" means not tested or data not available

Conclusion: The compound of the present invention has high exposure in plasma, low clearance rate, long half-life, high oral bioavailability, exhibits excellent pharmacokinetic properties, and is a good molecule for oral administration.

Experimental example 3 Anti-tumor activity test of the compounds of the present invention on animal tumor models in vivo

Purpose of the experiment : To investigate the antitumor effect of the compound of the present invention on the human prostate cancer LNCaP clone FGC xenograft tumor model. Experimental method:

Male CB17-SCID mice were subcutaneously inoculated with LNCaP clone FGC human prostate cancer cell line. On the 17th day after inoculation (the day of inoculation was regarded as day 0), they were randomly divided into operation group and non-operation group according to tumor volume and body weight. (6 animals) underwent castration. On the 20th day after inoculation, the animals in the non-operation group were randomly divided into the non-operation control group and the compound group (6 animals in each group) according to the tumor volume and body weight, and were given as follows: medicine treatment.

Group 1 (non-surgical control group): after inoculation on the 20th day of grouping, the drug was given in the afternoon of the same day, once a day, and the vehicle (5% DMSO + 40% PEG400 + 10% solutol+45% water).

Group 2 (castration surgery group): after inoculation on the 20th day of grouping, the drug was administered in the afternoon of the same day, once a day, and the vehicle (5%DMSO+40%PEG400+10%solutol +45% water).

Group 3 (Compound 1 ): After inoculation on the 20th day of grouping, administration was started in the afternoon of the same day, once a day, compound 1 was intragastrically administered at a dose of 100 mg/kg body weight, and the vehicle was 10% DMAC + 10% solutol + 80% saline.

During the experiment period, the tumor volume was measured twice a week and the body weight of the mice was weighed, and the tumor proliferation rate was calculated.

Calculation formula: tumor volume = length × width ² /2, tumor proliferation rate = tumor volume in the treatment group/tumor volume in the non-operative control group × 100%. The Student's t-test was used for statistical analysis between the groups, and p<0.05 was considered to be significantly different. Experimental results:

On the human prostate cancer LNCaP clone FGC xenograft tumor model, the tumor proliferation rate of the castration operation group was 69.52% on the 21st day of administration, and the tumor proliferation rate of compound 1 on the 21st day of administration was 37.09% (compared with the non-operative control group compared p<0.05). The specific results are shown in Table 3.

Table 3 Inhibitory results of compounds of the present invention on human prostate cancer LNCaP clone FGC xenograft tumor model group Tumor volume (mm ³ , Mean± SD, n=6) tumor growth rate Tumor volume on day 0 of administration Tumor volume at day 21 non-operative control group 175.83±24.18 559.96±123.35 - castration surgery group 155.74±36.42 389.27±111.99 69.52% Compound 1 176.01±20.42 207.69±42.83 37.09%

Conclusion: the compound of the present invention has remarkable antitumor effect.

none.

none.

Claims (15)

A compound represented by formula (II) or a pharmaceutically acceptable salt thereof,

Wherein, R ₁ is selected from -S(=O) _m -C _1-3 alkyl and -S(=O) _m -C _3-6 cycloalkyl, the -S(=O) _m -C _{1- 3} alkyl and -S(=O) _m -C _3-6 cycloalkyl are independently optionally substituted by 1, 2 or 3 R _a ; R ₂ and R ₃ are each independently selected from H, F, Cl , Br, I and C _1-3 alkyl, said C _1-3 alkyl is optionally substituted by 1, 2 or 3 R _b ; each R ₄ is independently selected from F, Cl, Br, I, C _1-3 alkoxy, -OC _3-6 cycloalkyl and C _3-6 cycloalkyl, said C _1-3 alkoxy, -OC _3-6 cycloalkyl and C _3-6 cycloalkyl are independently optionally substituted by 1, 2 or 3 R _c ; R ₈ is selected from -C(=O)OR ₆ ; R ₆ is selected from H and C _1-3 alkyl, the C _1-3 alkyl is optionally substituted by 1, 2 or 3 F; n is selected from 1, 2, 3 and 4; m is selected from 0; each R _a , R _b and R _c are independently selected from H, F, Cl, Br and I. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein R ₁ is selected from -S(=O) _m -C _1-3 alkyl, said -S(=O) _m -C _{1 -3} alkyl is optionally substituted with 1, 2 or 3 R _a . The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein R ₁ is selected from -SCH ₃ , -SCF ₃ , -SCH ₂ CH ₃ and -SCH ₂ CF ₃ . The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein R ₂ is selected from H, F, Cl and CH ₃ , and the CH ₃ is optionally substituted by 1, 2 or 3 Fs. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein R ₃ is selected from H, F, Cl and CH ₃ , and the CH ₃ is optionally substituted by 1, 2 or 3 Fs. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein R ₂ and R ₃ are each independently selected from H. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein each R ₄ is independently selected from F, Cl, -OCH ₃ , -OCH ₂ CH ₃ , -O-cyclopropyl, -O -cyclobutyl, cyclopropyl and cyclobutyl, said -OCH ₃ , -OCH ₂ CH ₃ , -O-cyclopropyl, -O-cyclobutyl, cyclopropyl and cyclobutyl are independently any Can be replaced by 1, 2 or 3 F's. The compound or a pharmaceutically acceptable salt thereof as claimed in item 7, wherein each R ₄ is independently selected from F, Cl, -OCH ₃ , -OCH ₂ CH ₃ ,

Cyclopropyl and Cyclobutyl. The compound or pharmaceutically acceptable salt thereof as claimed in item 8, wherein each R ₄ is independently selected from

, F and -OCH ₃ . The compound or pharmaceutically acceptable salt thereof as claimed in item 1 or 9, wherein the structural unit

selected from

and

The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein R ₈ is selected from -C(=O)OH. The compound or pharmaceutically acceptable salt thereof according to any one of claims 1 to 11, which is selected from the group consisting of,

Wherein, R ₁ , R ₂ , R ₃ , R ₄ and n are as defined in any one of claims 1 to 11. The compound or pharmaceutically acceptable salt thereof as claimed in item 12, which is selected from,

, where the structural unit

selected from

R ₅ is selected from C _1-3 alkyl, and the C _1-3 alkyl is optionally substituted by 1, 2 or 3 R _a ; R ₂ , R ₃ , R ₄ , n and R _a are as set forth in claim 12 definition. A compound selected from the following compounds or a pharmaceutically acceptable salt thereof:

and

. Application of a compound as described in any one of claims 1 to 14 or a pharmaceutically acceptable salt thereof in the preparation of drugs related to GnRH receptor antagonists.