Phosphoric acid and ester compound thereof and application thereof

Abstract

The invention relates to the technical field of medicines, in particular to phosphoric acid and ester compounds thereof, and application of the compounds in disease treatment medicines. The structural formula of the compound is as shown in the formula I, and the compound has the activity of inhibiting combination of PD-1 and PD-L1 and shows an excellent anti-tumor effect. # imgabs0 #

Description

Technical Field

[0001] The present invention relates to the field of medical technology, in particular to a phosphoric acid and its ester compound and application of the compound in disease treatment drugs. Background Art

[0002] Phosphate functional groups are widely present in living organisms, including genetic materials (including DNA and RNA), tissues and organs. Phosphate functional groups also exist in the structures of drugs that have been marketed or candidate drugs in the clinical research stage, such as:

[0003]

[0004] In recent years, although a series of PD-1 / PD-L1 inhibitors have been disclosed, there is still a need to discover and develop new compounds with better efficacy and pharmacokinetic properties. Summary of the invention

[0005] The primary purpose of the present invention is to provide a compound as shown in general formula I or II, which exhibits excellent activity in inhibiting the binding of PD-1 and PD-L1.

[0006] The technical solution adopted by the present invention is:

[0007] The present invention provides a compound as shown in formula I, its stereoisomers, tautomers or pharmaceutically acceptable salts,

[0008]

[0009] Wherein, Ring A and Ring B are each independently aryl, halogenated aryl, heteroaryl, halogenated heteroaryl, heterocyclic group, or halogenated heterocyclic group;

[0010] X1, X2 and X3 are each independently a nitrogen atom, CR X1 , CR X2 , CR X3 ;

[0011] Z is a carbon atom or a nitrogen atom; Y is an oxygen atom;

[0012] L is a linking group selected from the following groups:

[0013]

[0014] R A , R B , R X1 , R X2 , R X3 , R 1 , R 2 , R 3 , R4 , R 5 , R 6 , R 7 , R 8 , R 9 , R 10 Each is independently hydrogen, deuterium, halogen, alkyl, deuterated alkyl, haloalkane, cycloalkyl, hydroxy, alkoxy, deuterated alkyloxy, amino, carbonyl, sulfonyl, sulfone, phosphorus carbonyl, substituted silicon, alkenyl, alkynyl;

[0015] or R 1 , R 2 Together with the adjacent phosphorus atom, it forms a 5- to 8-membered substituted heterocyclic structure;

[0016] m = 0, 1, 2, 3, 4;

[0017] n = 0, 1, 2, 3, 4;

[0018] q=0, 1, 2, 3, 4.

[0019] The present invention provides a compound as shown in Formula II, its stereoisomers, tautomers or pharmaceutically acceptable salts,

[0020]

[0021] R A , R B , R X1 , R X2 , R 1 , R 2 , R 3 , R 4 , R 5 , R 6 , R 7 , R 8 , R 9 , R 10 , R 11 , R 12 , R 13 , R 14 , R 15 , R 16 , R 17 , R 18 , R 19 Each is independently hydrogen, deuterium, halogen, alkyl, deuterated alkyl, haloalkane, cycloalkyl, hydroxy, alkoxy, deuterated alkyloxy, amino, carbonyl, sulfonyl, sulfone, phosphorus carbonyl, substituted silicon, alkenyl, alkynyl;

[0022] or R 1 , R 2Together with the adjacent phosphorus atom, it forms a 5- to 8-membered substituted heterocyclic structure, wherein the substituted heterocyclic structure is selected from the following structures:

[0023]

[0024] The present invention proposes the following compounds, their stereoisomers, tautomers or pharmaceutically acceptable salts,

[0025]

[0026] The present invention provides a pharmaceutical composition comprising a therapeutically effective dose of the above compound or its stereoisomer, tautomer or pharmaceutically acceptable salt and a pharmaceutically acceptable carrier.

[0027] The present invention proposes the use of the above-mentioned compound or a pharmaceutically acceptable salt thereof in the preparation of a PD-1 / PD-L1 inhibitor drug.

[0028] The present invention provides the use of the above-mentioned compound, its pharmaceutically acceptable salt or pharmaceutical composition in the preparation of a drug for treating or preventing tumors. The tumor diseases are melanoma, lung cancer, liver cancer, basal cell carcinoma, kidney cancer, myeloma, bile duct cancer, brain cancer, breast cancer, cervical cancer, choriocarcinoma, colon cancer, rectal cancer, head and neck cancer, peritoneal tumors, fallopian tube cancer, endometrial cancer, esophageal cancer, gastric cancer, leukemia, lymphoma, sarcoma, neuroblastoma, oral cancer, ovarian cancer, pancreatic cancer, prostate cancer, testicular cancer, skin cancer and thyroid cancer.

[0029] The technical solution provided by the embodiment of the present invention has the following advantages compared with the prior art:

[0030] The present invention discloses a novel class of molecules containing "phosphoric acid and its esters" functional groups, which are compounds having good activity of inhibiting the binding of PD-1 and PD-L1.

[0031] The present invention also discloses a class of "phosphate" compounds, such as compound 6, which itself does not have the activity of inhibiting the binding of PD-1 and PD-L1, but its metabolites, such as compound 2, have the activity of inhibiting the binding of PD-1 and PD-L1. This class of "phosphate" compounds, such as compound 6, has the pharmacokinetic characteristics of liver distribution, and it is expected that this class of molecules has the potential to be a candidate molecule for the development of future "liver-targeted drugs". BRIEF DESCRIPTION OF THE DRAWINGS

[0032] Figure 1 After compound 6 was intragastrically administered to C57 mice, samples were collected at different time points to test the concentration of compound 2. DETAILED DESCRIPTION

[0033] In order to make the above-mentioned objects, features and advantages of the present invention more obvious and easy to understand, the specific implementation methods of the present invention are described in detail below in conjunction with the specification.

[0034] The compounds provided in the embodiments of the present invention can be prepared by standard synthetic methods known in the art, and this specification provides a general method for preparing the compounds of the present invention. The starting materials can usually be obtained commercially or prepared by methods well known to those skilled in the art.

[0035] All technical and scientific terms used in this specification have the meanings commonly understood by those of ordinary skill in the art.

[0036] Table 1:

[0037]

[0038] The term "alkyl" refers to a saturated aliphatic hydrocarbon group having 1 to 10 carbon atoms in this article, and the term includes straight and branched hydrocarbon groups. Non-limiting examples of alkyl include methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, neopentyl, n-hexyl, etc. Alkyl described herein can be optionally substituted with one or more of the following substituents: deuterium, fluorine, chlorine, bromine, iodine, cyano, nitro, hydroxyl, carboxyl, amino, alkyl, alkoxy, acyl, acyloxy, oxo, amide, ester, amine, cycloalkyl, cycloalkenyl, heterocycloalkyl, alkenyl, alkenyloxy, alkynyl, cycloalkyloxy, heterocycloalkyloxy, aryloxy, heteroaryloxy, aryl or heteroaryl.

[0039] The term "aryl" herein refers to a 6-10 membered all-carbon monocyclic or fused polycyclic (i.e., a ring sharing adjacent carbon atom pairs) group, a polycyclic (i.e., a ring with adjacent carbon atom pairs) group having a conjugated π electron system. The aryl group can be covalently linked to the defined chemical structure at any carbon atom that produces a stable structure. The aryl group described herein can be optionally substituted with one or more of the following substituents: fluorine, chlorine, bromine, iodine, cyano, nitro, hydroxyl, carboxyl, amino, alkyl, alkoxy, acyl, amide, ester, amine, sulfonyl, sulfinyl, cycloalkyl, cycloalkenyl, heterocycloalkyl, alkenyl, alkynyl and cycloalkoxy.

[0040] The term "heterocyclic group" refers to a ring system containing a nitrogen atom or an oxygen atom, and the ring system can be "connected" to aromatic and non-aromatic ring systems, or linked to other ring systems through a "spiro carbon atom".

[0041] The term "heteroaryl" as used herein refers to an aromatic group consisting of 5 to 10 atoms and containing at least one heteroatom selected from N, O or S. The term may have a single ring (non-limiting examples include furan, thiophene, imidazole, pyrazole, pyridine, pyrazine, oxazole, thiazole, etc.) or multiple fused rings (non-limiting examples include benzothiophene, benzofuran, indole, isoindole, etc.), wherein the fused rings may or may not be aromatic groups containing heteroatoms, provided that the point of attachment is through the atoms of the aromatic heteroaryl group. The heteroaryl groups described herein may be optionally substituted with one or more of the following substituents: fluorine, chlorine, bromine, iodine, cyano, nitro, hydroxy, amino, alkyl, alkoxy, acyl, acyloxy, amide, ester, amine, sulfonyl, sulfinyl, cycloalkyl, cycloalkenyl, heterocycloalkyl, alkenyl, alkynyl and cycloalkoxy.

[0042] The term "alkenyl" refers to an alkenyl group having 2 to 8 carbon atoms and having at least one alkenyl unsaturated site in this article. The limiting examples of alkenyl include vinyl, propenyl, allyl, isopropenyl, butenyl, isobutenyl etc. Alkenyl described herein can be optionally substituted with one or more of the following substituents: deuterium, fluorine, chlorine, bromine, iodine, cyano, nitro, hydroxyl, carboxyl, amino, alkyl, alkoxy, acyl, amide, ester, amine, sulfonyl, sulfinyl, cycloalkyl, cycloalkenyl, heterocycloalkyl, cycloalkyloxy, sulfydryl, alkyl sulfydryl, deuterated alkyl sulfydryl, sulfone, sulfoxide, amino, silicon-based, phosphono, deuterated alkyl, heterocycloalkyl, aryl, heteroaryl, alkynyl, alkenyl, arylalkyl, ester.

[0043] The term "alkynyl" refers herein to an alkyl group in which two adjacent carbon atoms are connected by a triple bond, wherein the alkyl group is as defined herein. Alkynyl refers to an unsaturated alkyl group as defined above consisting of at least two carbon atoms and at least one carbon-carbon triple bond, such as ethynyl, 1-propynyl, 2-propynyl, 1-, 2- or 3-butynyl, etc. Alkynyl can be substituted or unsubstituted, and when substituted, substituents are preferably one or more of the following groups, independently selected from deuterium, fluorine, chlorine, bromine, iodine, cyano, nitro, hydroxyl, carboxyl, amino, alkyl, alkoxy, acyl, amide, ester, amine, sulfonyl, sulfinyl, cycloalkyl, cycloalkenyl, heterocycloalkyl, cycloalkyloxy, sulfhydryl, alkylsulfhydryl, deuterated alkylsulfhydryl, sulfone, sulfoxide, amino, silicon, phosphonyl, deuterated alkyl, heterocycloalkyl, aryl, heteroaryl, alkynyl, alkenyl, arylalkyl, ester.

[0044] The term "pharmaceutically acceptable carrier" herein refers to a non-toxic solid, semi-solid or liquid filler, diluent, adjuvant, packaging material or other formulation excipient. The carrier used may be adapted to the corresponding administration form, and may be formulated into injection, (injection) lyophilized powder, spray, oral solution, oral suspension, tablet, capsule, enteric-coated tablet, pill, powder, granule, sustained release or delayed release formulations using carriers known to those skilled in the art.

[0045] The terms "prevention" and "treatment" herein have the meanings conventionally understood by those skilled in the art, i.e., "prevention" refers to the administration of a pharmaceutical composition or active compound before the onset of a disease or before the onset of symptoms to prevent, delay and / or alleviate the onset of a corresponding disease or the onset of symptoms; "treatment" refers to the administration of a pharmaceutical composition or active compound at the time of or after the onset of a disease, or at the time of or after the onset of symptoms, to eliminate the corresponding condition, alleviate the severity of the corresponding symptoms, or delay the development of the corresponding disease.

[0046] The term "subject" herein preferably refers to a mammal, especially a human or an experimental animal (such as a mouse, rat or rabbit, etc.).

[0047] The term "effective amount" herein refers to the appropriate dose of a drug or treatment method required to produce the desired effect. Among them, a therapeutically effective dose refers to a dose of a drug that can treat a disease or symptom, while a preventive effective dose refers to a dose of a drug or measure that can prevent the occurrence of a disease or reduce the risk of a disease. Those skilled in the art can determine the effective dose based on specific factors, such as individual age, weight, severity of the disease, etc.

[0048] The present invention provides a compound with a structural formula as shown in Formula I, its stereoisomers, tautomers or pharmaceutically acceptable salts,

[0049]

[0050] Wherein, Ring A and Ring B are each independently aryl, halogenated aryl, heteroaryl, halogenated heteroaryl, heterocyclic group, or halogenated heterocyclic group;

[0051] X1, X2 and X3 are each independently a nitrogen atom, CR X1 , CR X2 , CR X3 ;

[0052] Z is a carbon atom or a nitrogen atom; Y is an oxygen atom;

[0053] L is a linking group selected from the following groups:

[0054]

[0055] R A , RB , R X1 , R X2 , R X3 , R 1 , R 2 , R 3 , R 4 , R 5 , R 6 , R 7 , R 8 , R 9 , R 10 Each is independently hydrogen, deuterium, halogen, alkyl, deuterated alkyl, haloalkane, cycloalkyl, hydroxy, alkoxy, deuterated alkyloxy, amino, carbonyl, sulfonyl, sulfone, phosphorus carbonyl, substituted silicon, alkenyl, alkynyl;

[0056] or R 1 , R 2 Together with the adjacent phosphorus atom, it forms a 5- to 8-membered substituted heterocyclic structure;

[0057] m = 0, 1, 2, 3, 4;

[0058] n = 0, 1, 2, 3, 4;

[0059] q=0, 1, 2, 3, 4.

[0060] In the embodiments of the present invention, compounds of the present invention having the same molecular formula but different properties or sequences of their atomic bonds or different spatial arrangements of their atoms are referred to as "isomers". Isomers with different spatial arrangements of atoms are referred to as "stereoisomers". All isomers of the compounds described herein (e.g., cis-, trans-, or diastereomers) alone or in any mixture thereof are included within the scope of the embodiments of the present invention. All of these forms include enantiomers, diastereomers, cis, trans, syn, anti, tautomers, and mixtures thereof. Stereoisomeric mixtures (e.g., mixtures of diastereomers) can be separated into their corresponding isomers in a known manner using suitable separation methods.

[0061] In an embodiment of the present invention, the pharmaceutically acceptable salts of the compound of Formula I may include acid addition salts and basic salts. Suitable acid addition salts are formed by acids that can form non-toxic salts. Suitable basic salts are formed by bases that form non-toxic salts. Examples include salts of aluminum, arginine, N-benzathine, calcium, choline, diethylamine, diolamine, glycine, lysine, magnesium, meglumine, ethanolamine, potassium, sodium, tromethamine, and zinc.

[0062] As a preferred technical solution of the embodiment of the present invention, Ring A and Ring B are each independently selected from an aryl group or a halogenated aryl group, and further independently selected from a phenyl group or a halogenated phenyl group. Further optionally, Ring A and Ring B are each independently a halogenated aryl group. The halogenated group may be monohalogenated or polyhalogenated.

[0063] As a preferred technical solution of the embodiment of the present invention, X1 and X2 are each independently -CR X1 =, -CR X2 =, R X1 , R X2 Each is independently hydrogen, deuterium, halogen, alkyl, deuterated alkyl, halogenated alkane. Further optionally, X1 and X2 are both -CH=.

[0064] As a preferred technical solution of an embodiment of the present invention, X3 is a nitrogen atom.

[0065] As a preferred technical solution of an embodiment of the present invention, Z is a nitrogen atom.

[0066] As a preferred technical solution of the embodiment of the present invention, R 1 , R 2 are independently hydroxyl, alkoxy, or R 1 , R 2 Together with the adjacent phosphorus atom, it forms a 5- to 8-membered substituted heterocyclic structure.

[0067] As a preferred technical solution of the embodiment of the present invention, R 3 , R 4 , R 5 , R 6 , R 7 , R 8 , R 9 , R 10 Each is independently hydrogen, deuterium, halogen, alkyl, deuterated alkyl, or halogenated alkane. 3 , R 4 , R 5 , R 6 , R 7 , R 8 , R 9 , R 10 All are hydrogen.

[0068] The present invention also provides a compound having a structural formula as shown in Formula II, a stereoisomer, a tautomer or a pharmaceutically acceptable salt thereof,

[0069]

[0070] Among them, R A , R B, R X1 , R X2 , R 1 , R 2 , R 3 , R 4 , R 5 , R 6 , R 7 , R 8 , R 9 , R 10 , R 11 , R 12 , R 13 , R 14 , R 15 , R 16 , R 17 , R 18 , R 19 Each is independently hydrogen, deuterium, halogen, alkyl, deuterated alkyl, haloalkane, cycloalkyl, hydroxy, alkoxy, deuterated alkyloxy, amino, carbonyl, sulfonyl, sulfone, phosphorus carbonyl, substituted silicon, alkenyl, alkynyl;

[0071] or R 1 , R 2 Together with the adjacent phosphorus atom, it forms a 5- to 8-membered substituted heterocyclic structure, wherein the substituted heterocyclic structure is selected from the following structures:

[0072]

[0073] m = 0, 1, 2, 3, 4;

[0074] n = 0, 1, 2, 3, 4;

[0075] q=0, 1, 2, 3, 4.

[0076] As a preferred technical solution of the embodiment of the present invention, R A , R B , R X1 , R X2 , R 1 , R 2 , R 3 , R 4 , R 5 , R 6 , R 7 , R 8 , R 9 , R 10 , R 11 , R 12 , R 13 , R 14 , R 15 , R 16 , R 17 , R18 , R 19 Each is independently hydrogen, deuterium, halogen, hydroxyl, amino, sulfone, phosphorus carbonyl, carbonamide, sulfonamide, C1-C6 alkyl, deuterated C1-C6 alkyl, halogenated C1-C6 alkane, C3-C8 cycloalkyl, C1-C6 alkoxy, deuterated C1-C6 alkyloxy, substituted silicon, C2-C8 alkenyl, or C2-C8 alkynyl.

[0077] As a preferred technical solution of the embodiment of the present invention, R 11 , R 12 , R 13 , R 14 At least one of the substituents is halogen; further optionally, R 14 is halogen; further optionally, R 14 is a halogen, and R 11 , R 12 , R 13 All are hydrogen atoms.

[0078] As a preferred technical solution of the embodiment of the present invention, R 15 , R 16 , R 17 , R 18 , R 19 At least one of the substituents is halogen; further optionally, R 19 is halogen; further optionally, R 19 is halogen, R 15 , R 16 , R 17 , R 18 All are hydrogen atoms.

[0079] As a preferred technical solution of the embodiment of the present invention, R 1 , R 2 Each independently hydroxyl, C1-C6 alkoxy or R 1 , R 2 Together with the adjacent phosphorus atom, it forms a substituted heterocyclic structure.

[0080] As a preferred technical solution of the embodiment of the present invention, R X1 , R X2 Each is independently hydrogen, deuterium, or halogen. X1 is hydrogen. Further optionally, R X2 For hydrogen.

[0081] As a preferred technical solution of the embodiment of the present invention, R A , R B are independently hydrogen or deuterium. A is hydrogen. Further optionally, RB For hydrogen.

[0082] As a preferred technical solution of the embodiment of the present invention, m=1.

[0083] As a preferred technical solution of the embodiment of the present invention, n=1.

[0084] As a preferred technical solution of the embodiment of the present invention, q=1 or 2.

[0085] The present invention also provides the following compounds, their stereoisomers, tautomers or pharmaceutically acceptable salts,

[0086]

[0087] The present invention also proposes a pharmaceutical composition, which includes a therapeutically effective dose of the above-mentioned compound or its stereoisomer, tautomer or pharmaceutically acceptable salt and a pharmaceutically acceptable carrier. The pharmaceutical composition of the present invention can be prepared by methods known in the art. The pharmaceutical composition used in the present invention can be prepared using one or more pharmaceutically acceptable carriers according to conventional methods, and the carrier includes excipients and adjuvants, which are conducive to processing the active compound into a pharmaceutically usable preparation. Suitable preparations depend on the selected route of administration. Pharmaceutically acceptable excipients and carriers are generally known to those skilled in the art and are therefore included in the scope of the present invention. The pharmaceutical composition of the embodiment of the present invention can be in various forms, such as tablets, capsules, powders, syrups, solutions, suspensions and aerosols, etc., and the compounds of the embodiment of the present invention can be present in suitable solid or liquid carriers or diluents. The pharmaceutical composition of the present invention can also be stored in suitable sterile equipment for injection or infusion. Usually, the amount of active ingredient in the pharmaceutical composition is 0.01% by weight to 99% by weight of the pharmaceutical composition.

[0088] The compounds of the embodiments of the present invention or pharmaceutical compositions containing the compounds of the embodiments of the present invention can be used clinically in mammals (including humans) through oral, nasal, skin, lung or gastrointestinal administration routes. The preferred route of administration is oral. The preferred daily dose is 0.5-200 mg / kg body weight, taken once or in divided doses. Regardless of the method of administration, the optimal dose for an individual should be determined according to the specific treatment. Usually, it is started with a small dose and the dose is gradually increased until the most suitable dose is found. The effective dose of the active ingredient used may vary with the compound used, the mode of administration and the severity of the disease to be treated. However, generally, when the compound of the present invention is administered at a dose of about 1-300 mg / kg animal body weight per day, satisfactory effects can be obtained, preferably administered in 1-3 divided doses per day, or administered in a sustained release form. For most large mammals, the total daily dose is about 5-1000 mg, preferably about 10-500 mg. The dosage form suitable for oral administration contains about 1-200 mg of the active compound intimately mixed with a solid or liquid pharmaceutically acceptable carrier. This dosage regimen may be adjusted to provide the optimum therapeutic response. For example, several divided doses may be administered daily, or the dose may be proportionally reduced as required by the exigencies of the therapeutic situation.

[0089] The embodiments of the present invention also provide methods for treating or preventing a disease or condition, which include administering an effective amount of a compound comprising an embodiment of the present invention or a pharmaceutically acceptable salt thereof to an individual. The compound of the present invention or its pharmaceutical composition can be administered orally, parenterally, topically, rectally, mucosally or intestinally. Parenteral administration includes indirect injection to produce systemic effects or direct injection to the diseased site. Topical administration includes treatment of the skin or organs (e.g., eyes or ears) that can be easily reached by topical application. Transdermal delivery is also included to produce systemic effects. Rectal administration includes the form of suppositories. Preferred routes of administration are oral and parenteral.

[0090] The present invention also provides the use of the above-mentioned compound or its pharmaceutically acceptable salt in the preparation of PD-1 / PD-L1 inhibitor drugs. Experiments have confirmed that the compound of the present invention has good activity in inhibiting the binding of PD-1 and PD-L1.

[0091] The present invention also provides the use of the above-mentioned compound, its pharmaceutically acceptable salt or pharmaceutical composition in the preparation of a drug for treating or preventing tumors. Wherein, the tumor disease is melanoma, lung cancer, liver cancer, basal cell carcinoma, kidney cancer, myeloma, biliary tract cancer, brain cancer, breast cancer, cervical cancer, choriocarcinoma, colon cancer, rectal cancer, head and neck cancer, peritoneal tumor, fallopian tube cancer, endometrial cancer, esophageal cancer, gastric cancer, leukemia, lymphoma, sarcoma, neuroblastoma, oral cancer, ovarian cancer, pancreatic cancer, prostate cancer, testicular cancer, skin cancer and thyroid cancer.

[0092] The compounds of the present invention and corresponding preparation methods are further explained and listed below by examples and preparation. It should be understood that although typical or preferred reaction conditions are given in the specific examples, other reaction conditions can also be used by those skilled in the art. Optimum reaction conditions can vary with the specific reaction substrate or solvent used, but the conditions can be determined by conventional optimization by those skilled in the art.

[0093] Intermediate preparation

[0094] Intermediate 1:

[0095]

[0096] To a 2L four-necked flask, add SM-1 (94.77g), SM-2 (71.85g), K2CO3 (147.85g), 1,4-dioxane (1L), and water (100mL), stir, replace with nitrogen once, add Pd(dppf)Cl2 (15.65g), replace with nitrogen three times, increase the temperature to 80°C, react for 3 hours, and take samples every hour for detection until SM-I is consumed. The reaction solution temperature was reduced to 40°C, passed through a diatomaceous earth funnel (150 mm, 100 g), and the diatomaceous earth was rinsed 4 times with ethyl acetate (300 mL + 300 mL + 200 mL + 200 mL). The filtrate and washings were combined, washed once with 10% NaCl (1 L), and then washed once with saturated brine (1 L). The organic phase was collected, dried with anhydrous Na2SO4 (200 g) for 15 min, filtered, the filtrate was collected, and concentrated to dryness by rotary evaporation to obtain a black oil (124 g).

[0097] Intermediate 2:

[0098]

[0099] Add intermediate 1 (124 g), SM-3 (97.68), t-Buxphos (40.45 g), Cs2CO3 (387.97 g), toluene (1.24 L) to a 2L four-necked bottle, stir, replace with nitrogen once, add Pd(OAc)2 (10.69 g), replace with nitrogen three times, heat up, target 80°C, react for 5 hours, sample and detect every 1 h until SM-3 is consumed, the reaction liquid temperature drops to 40°C, pass through a diatomaceous earth funnel (150 mm, 100 g), rinse the diatomaceous earth with EA 5 times (300 mL × 5), and concentrate the filtrate and the first rinse on a rotary evaporator Dry, add 2 to 5 times of eluent to the dried oil, wash once with 10% NaCl (1L), then wash once with saturated brine (1L), pass the emulsified layer through a diatomaceous earth funnel, collect the organic phase, warm to 50°C, add activated carbon (10g), anhydrous Na2SO4 (200g), stir for 1h, filter, collect the filtrate, cool below 10°C, slowly add methanesulfonic acid (45.78g) dropwise, stir at room temperature for 1h, filter, collect the filter cake, beat with ethyl acetate, stir at 40-45°C for 1h, naturally cool to 25°C, filter, collect the filter cake, and dry at 40°C for 5h to obtain a crude intermediate 2 (154g).

[0100] Add the crude intermediate 2 (154 g) and DMF (385 mL) into a 1L three-necked flask and stir. Heat to 35°C, add dichloromethane (310 mL) and activated carbon (15.4 g), stir for 1 h, filter, rinse the filter cake with N,N-dimethylformamide: dichloromethane = 5:4 (92 mL), collect the filtrate and washings, heat to 35°C, add ethyl acetate (1055 mL) dropwise, cool naturally to 27°C after the addition, cool to below 10°C in an ice bath, stir for 2 h, filter, collect the filter cake, and dry naturally to obtain intermediate 2 (94 g). 1 H NMR (500MHz, DMSO-d6) δ: 9.09 (s, 2H), 7.64-6.86 (m, 9H), 5.46 (s, 2H), 4.25 (s, 2H), 3.47 (t, 2H), 3.00 (t, 2H), 2.34 (s, 3H).

[0101] Compound preparation

[0102] Embodiment 1:

[0103]

[0104] In a 25 mL single-mouth bottle, add intermediate 2 (200 mg), add 1 mL of water, 2 mL of tetrahydrofuran, stir to dissolve, add triethylamine (99.8 mg), add diethyl vinyl phosphate (117 mg), react at room temperature for 48 h, extract with 5 mL and 2 mL of ethyl acetate respectively, combine the organic phases, wash with saturated sodium chloride, dry with anhydrous sodium sulfate, concentrate to dryness, and purify with a silica gel column to obtain 170 mg of a white solid. m / z: 533.40 (M+1). 1 H NMR(600MHz,DMSO)δ7.65(dd,1H),7.53–7.48(m,1H),7.48–7.41(m,2H),7.38(dd,2H),7.32(d,2H),6.72(d,1H), 5.44(s,2H),3.99(ddd,4H),3.52(s,2H),2.79(d,2H),2.74(d,2H),2.69(d,2H),2.11–1.99(m,2H),1.22(t,6H).

[0105] Embodiment 2:

[0106]

[0107] Compound 1 (0.5 g) was added to a 25 mL single-mouth bottle, acetonitrile (5 mL) was added, trimethylsilyl bromide (0.73 g) was added, the temperature was raised to 80°C for reaction for 6 h, the system was concentrated to dryness, 2 mL of water was added, a solid was precipitated, the pH was adjusted to 10-12 with sodium hydroxide solution, extracted with 5 mL of methyl tert-butyl ether, the organic phase was discarded, the aqueous phase was adjusted to pH 3-4 with 1 mol / L hydrochloric acid, the system became an oil, the pH was adjusted to 9-10 with 8% sodium carbonate solution, solid precipitated after stirring, filtered, the solid was dissolved with methanol, concentrated to dryness, and slurried with 10 mL of ethyl acetate to obtain 70 mg of a khaki solid. m / z: 477.10 (M+1). 1 H NMR(600MHz,DMSO)δ11.71(s,1H),7.68(dd,1H),7.60(d,1H),7.55–7.44(m,2H),7.39(ddd,2H),7.36–7.28(m,2H ),6.88(d,1H),5.47(d,2H),4.50(s,1H),4.27(s,1H),3.78(s,1H),3.48–3.22(m,4H),2.98(s,1H),2.26(s,2H).

[0108] Embodiment 3:

[0109]

[0110] In a 25 mL single-mouth bottle, add intermediate 2 (100 mg), add 1 mL of water, 2 mL of tetrahydrofuran, stir to dissolve, add triethylamine (99.8 mg), add vinyl cyclic phosphoric acid diester SM-6 (140 mg), react at room temperature for 48 h, extract with 6 mL and 3 mL of ethyl acetate respectively, combine the organic phases, wash with saturated sodium chloride, dry with anhydrous sodium sulfate, concentrate to dryness, and purify with a preparative flash silica gel column to obtain 33 mg of a white solid. m / z: 613.11 (M+1). 1 H NMR(600MHz,DMSO)δ11.71(s,1H),7.68(dd,1H),7.60(d,1H),7.55–7.44(m,2H),7.39(ddd,2H),7.36–7.28(m,2H ),6.88(d,1H),5.47(d,2H),4.50(s,1H),4.27(s,1H),3.78(s,1H),3.48–3.22(m,4H),2.98(s,1H),2.26(s,2H).

[0111] Biological Testing

[0112] Experimental Example 1

[0113] This experimental example uses a PD-1 / PD-L1 binding assay kit (CISBIO) to detect the activity of the Formula I compound inhibitor PD-1 / PD-L1 interaction.

[0114] Experimental Materials: The kits were purchased from CISBIO: PD-1 / PD-L1 binding assay kits; 96-well plates: purchased from CISBIO.

[0115] Test equipment: Perkin Elmer, Model: EnVision.

[0116] Test compound: Dissolve in DMSO and dilute with diluent buffer; the DMSO concentration should not exceed 0.5%.

[0117] Experimental process:Use PD-1 / PD-L1 binding assay kits. Set up negative group, positive group and drug administration group, with 2 replicates in each group. For the positive control group, add 2μL diluent to the 96-well plate; 4μL PD-L1 and 4μL PD-1 diluted according to the instructions; for the negative control group, add 6μL diluent and 4μL PD-L1 to the 96-well plate; for the drug administration group, add 2μL of the test compound (I), 4μL PD-L1 and 4μL PD-1 to the 96-well plate in sequence. Seal the plate with a sealing film, centrifuge at 1000rpm for 1 minute, and incubate at room temperature for 15 minutes. Then add Anti-Tag-Eu3 diluted in Buffer + Mix equal volumes of Anti-tag-XL665, then add 10 μL of the mixture to each well, seal the plate, centrifuge at 1000 rpm for 1 minute, and incubate at room temperature for 2 hours. Remove the sealing film, use EnVision to read the fluorescence intensity at 665 nm and 615 nm, and calculate. Use Graphpad to calculate the IC value of the compound 50 .

[0118] The test results are shown in Table 2:

[0119] Table 2: Blocking effect of the compounds of the present invention on hPD-1 / hPD-L1

[0120] Example Compound No. <![CDATA[IC 50 (nM)]]> 1 56.6 2 15.8 6 >1000

[0121] The above data indicate that compounds 1 and 2 of the present invention have significant inhibitory activity on PD-1 / PD-L1 protein-protein interaction.

[0122] Experimental Example 2

[0123] This experimental example is used to evaluate the distribution of the test compound in major organs after a single intragastric bolus administration.

[0124] Route of administration: Compound 2 and compound 6 were administered by oral gavage;

[0125] Dosage: 5mg / kg; 5mL / kg.

[0126] Dosing frequency: Single dose.

[0127] Blood and organ collection:

[0128] At each blood collection point, 0.25-0.3 mL of systemic blood was collected into an EDTA-K2 anticoagulant tube, which was gently inverted to mix and then placed on crushed ice for storage and transportation; collection plan: PO blood collection point: 0.5, 1h, 2h, 4h, and 6h after administration; the animals were euthanized, and tissues and organs were collected, including the liver (0.3-0.5 g was collected from a fixed position in the liver lobe), kidney, and heart, which were weighed, placed in a 5 mL centrifuge tube, quickly frozen with dry ice, and stored at -20°C.

[0129] Sample preparation:

[0130] Solvent (methanol:acetonitrile:water at a ratio of 25:25:50) was added and the slurry was ground and homogenized. This process was quickly performed using a tissue grinder.

[0131] For liver, add solvent to 3 mL, add 1 large steel ball and 1 small steel ball, grind at 50 Hz for 60 seconds. Vortex to mix, centrifuge at 8000 rpm for 5 minutes. Take 0.3 mL of supernatant, add solvent to 3 mL, vortex to mix, centrifuge at 8000 rpm for 5 minutes. Take 0.1 mL of supernatant, add 0.2 mL of acetonitrile, vortex for 30 seconds, centrifuge at 10000 rpm for 5 minutes, take supernatant, and send for analysis of drug content.

[0132] Kidney, divided into two centrifuge tubes, add solvent to 3mL, add 1 large steel ball and 1 small steel ball, grind at 50Hz for 60s, observe the grinding effect, and grind multiple times. After grinding, vortex to mix, centrifuge at 8000rpm for 5min. Take 0.3mL of supernatant, add solvent to the same 3mL, vortex to mix, centrifuge at 8000rpm for 5min. Take 0.1mL of supernatant, add 0.2mL of acetonitrile, vortex for 30s, centrifuge at 10000rpm for 5min, take supernatant, and send for analysis of drug content.

[0133] For plasma, take 0.1 mL of plasma, add 0.2 mL of acetonitrile, vortex for 30 s, centrifuge at 10,000 rpm for 5 min, take the supernatant, and send it for analysis of drug content.

[0134] Detection method:

[0135] Chromatographic column: C18 Xtimate C18 (4.6×100mm) 341# with a guard column of the same model;

[0136] Mobile phase: A: 0.1% formic acid solution; B: acetonitrile;

[0137] The elution method adopts gradient elution, and the specific method is shown in Table 3:

[0138] Table 3

[0139] Time min A％ B％ 0 90 10 3 55 45 6 10 90 10 10 90 10.1 90 10 12.5 90 10

[0140] Flow rate: 0.4 mL / min;

[0141] Column temperature: 35°C;

[0142] Injection volume: 10 μL.

[0143] The experimental results are shown in Table 4:

[0144] Table 4

[0145] Blood (ng / mL) Kidney (ng / mL) Liver (ng / g) Liver / blood concentration ratio Liver / kidney concentration ratio Compound 2 7352 17182 16372 2.22 0.95 Compound 6 4891 13538 62643 12.8 4.62

[0146] According to the test data, the tissue distribution data bar chart of compound 6 is summarized as follows Figure 1 . The experimental results show that compared with compound 2, compound 6 of the present invention has a significant advantage in liver distribution. For example, the liver / blood concentration ratio (12.2) and liver / kidney concentration ratio (4.62) of compound 6 are greater than the corresponding concentration ratios of compound 2 (2.22 and 0.95). Compound 6 is a prodrug of compound 2. After animals are gavaged with compound 6, compound 2 can be concentrated in the liver, reducing the systemic exposure of compound 2 and avoiding potential systemic toxicity.

[0147] The above description is only a specific embodiment of the present invention, so that those skilled in the art can understand or implement the present invention. Various modifications to these embodiments will be apparent to those skilled in the art, and the general principles defined herein may be implemented in other embodiments without departing from the spirit or scope of the present invention. Therefore, the present invention will not be limited to the embodiments described herein, but should conform to the widest scope consistent with the principles and novel features disclosed herein.

Claims

1. A compound as shown in formula I, its stereoisomers, tautomers or pharmaceutically acceptable salts, in, Ring A and Ring B are each independently aryl, halogenated aryl, heteroaryl, halogenated heteroaryl, heterocyclic group, or halogenated heterocyclic group; X1, X2 and X3 are each independently a nitrogen atom, CR X1 , CR X2 , CR X3 ; Z is a carbon atom or a nitrogen atom; Y is an oxygen atom; L is a linking group selected from the following groups: R A , R B , R X1 , R X2 , R X3 , R 1 , R 2 , R 3 , R 4 , R 5 , R 6 , R 7 , R 8 , R 9 , R 10 Each is independently hydrogen, deuterium, halogen, alkyl, deuterated alkyl, haloalkane, cycloalkyl, hydroxy, alkoxy, deuterated alkyloxy, amino, carbonyl, sulfonyl, sulfone, phosphorus carbonyl, substituted silicon, alkenyl, alkynyl; or R 1 , R 2 Together with the adjacent phosphorus atom, it forms a 5- to 8-membered substituted heterocyclic structure; m＝0、1、2、3、4； n＝0、1、2、3、4； q＝0、1、2、3、4。 2. A compound of formula II, its stereoisomers, tautomers or pharmaceutically acceptable salts, R A , R B , R X1 , R X2 , R 1 , R 2 , R 3 , R 4 , R 5 , R 6 , R 7 , R 8 , R 9 , R 10 , R 11 , R 12 , R 13 , R 14 , R 15 , R 16 , R 17 , R 18 , R 19 Each is independently hydrogen, deuterium, halogen, alkyl, deuterated alkyl, haloalkane, cycloalkyl, hydroxy, alkoxy, deuterated alkyloxy, amino, carbonyl, sulfonyl, sulfone, phosphorus carbonyl, substituted silicon, alkenyl, alkynyl; or R 1 , R 2 Together with the adjacent phosphorus atom, it forms a 5- to 8-membered substituted heterocyclic structure, wherein the substituted heterocyclic structure is selected from the following structures: m＝0、1、2、3、4； n＝0、1、2、3、4； q＝0、1、2、3、4。 3. The compound according to claim 2 or its stereoisomer, tautomer, or pharmaceutically acceptable salt, characterized in that: R A , R B , R X1 , R X2 , R 1 , R 2 , R 3 , R 4 , R 5 , R 6 , R 7 , R 8 , R 9 , R 10 , R 11 , R 12 , R 13 , R 14 , R 15 , R 16 , R 17 , R 18 , R 19 Each is independently hydrogen, deuterium, halogen, hydroxyl, amino, sulfone, phosphorus carbonyl, carbonamide, sulfonamide, C1-C6 alkyl, deuterated C1-C6 alkyl, halogenated C1-C6 alkane, C3-C8 cycloalkyl, C1-C6 alkoxy, deuterated C1-C6 alkyloxy, substituted silicon, C2-C8 alkenyl, or C2-C8 alkynyl.

4. The compound according to claim 2 or its stereoisomer, tautomer, or pharmaceutically acceptable salt, characterized in that: R 11 , R 12 , R 13 , R 14 At least one of the substituents is a halogen; Optionally, R 15 , R 16 , R 17 , R 18 , R 19 At least one of the substituents is a halogen; Optionally, R 14 and R 19 is a halogen; Optionally, R 1 , R 2 Each independently hydroxyl, C1-C6 alkoxy or R 1 , R 2 Together with the adjacent phosphorus atom, they form the substituted heterocyclic structure.

5. The compound according to claim 2 or its stereoisomer, tautomer, or pharmaceutically acceptable salt, characterized in that: R X1 , R X2 Each is independently hydrogen, deuterium, or halogen; Optionally, R A , R B each independently hydrogen, deuterium; Optionally, m=1; Optionally, n=1; Optionally, q=1 or 2.

6. The following compounds, their stereoisomers, tautomers or pharmaceutically acceptable salts, 7. A pharmaceutical composition comprising a therapeutically effective dose of the compound according to any one of claims 1 to 6 or its stereoisomer, tautomer or pharmaceutically acceptable salt and a pharmaceutically acceptable carrier.

8. Use of a compound according to any one of claims 1 to 6 or a pharmaceutically acceptable salt thereof in the preparation of a PD-1 / PD-L1 inhibitor drug.

9. Use of the compound according to any one of claims 1 to 6 or a pharmaceutically acceptable salt thereof or the pharmaceutical composition according to claim 7 in the preparation of a medicament for treating or preventing tumors.

10. The use according to claim 9, characterized in that The tumor diseases are melanoma, lung cancer, liver cancer, basal cell carcinoma, kidney cancer, myeloma, biliary tract cancer, brain cancer, breast cancer, cervical cancer, choriocarcinoma, colon cancer, rectal cancer, head and neck cancer, peritoneal tumors, fallopian tube cancer, endometrial cancer, esophageal cancer, gastric cancer, leukemia, lymphoma, sarcoma, neuroblastoma, oral cancer, ovarian cancer, pancreatic cancer, prostate cancer, testicular cancer, skin cancer and thyroid cancer.

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