CN114524812A - Crystal form preparation and synthesis method of 1, 4-dihydro-1, 6-naphthyridine compound - Google Patents

Abstract

The invention relates to a synthesis and purification method for preparing a compound I ((4S) -4- (4-cyano-2-methoxyphenyl) -5-ethoxy-2, 8-dimethyl-1, 4-dihydro-1, 6-naphthyridine-4-deuterium-3-formamide). And also relates to a crystal form A and a crystal form B of the compound I and a preparation method thereof. The improved process for the synthesis of compounds (IX), (X) and (R, S) - (I) according to the invention is convenient for large scale operation, and the invention provides an alternative to the separation of enantiomeric mixtures to prepare highly pure chiral compounds I, which can be operated using conventional pilot plant equipment (stirred vessel/separation apparatus) without the need to purchase additional special equipment, overcoming the problems of low yield and high cost, difficulty in obtaining the compounds of the original published synthetic routePurification and difficult large-scale production.

Description

Crystal form preparation and synthesis method of 1, 4-dihydro-1, 6-naphthyridine compound

Technical Field

The invention relates to a crystal form and preparation of a crystal form of a 1, 4-dihydro-1, 6-naphthyridine compound (4S) -4- (4-cyano-2-methoxyphenyl) -5-ethoxy-2, 8-dimethyl-1, 4-dihydro-1, 6-naphthyridine-4-deuterium-3-formamide, and a synthesis and purification method of the compound. More particularly, the present invention relates to a method for preparing the pharmaceutically active ingredient compounds, a pharmaceutical composition comprising the same, and a Mineralocorticoid (MR) receptor modulator using the same for improving vascular function, and to their utility potentially in the treatment and/or prevention of clinical conditions including cardiovascular and metabolic diseases, such as hypertension, heart failure, chronic kidney disease, diabetic nephropathy, fibrosis, endothelial dysfunction, and muscle atrophy.

Background

The Mineralocorticoid Receptor (MR) is a member of the steroid receptor family, which regulates blood pressure by modulating the effect of aldosterone (Aldo) on renal sodium treatment. Over the past decade, the expression of MR in the cardiovascular system has become clear, and there is increasing interest in understanding the direct role of MR in regulating vascular function and promoting cardiovascular disease. This interest has resulted from a number of clinical studies in which drugs that reduce MR activation also reduce the incidence of heart disease, stroke, and mortality. The presence of functional mineralocorticoid receptors in vascular smooth muscle and endothelial cells has been demonstrated, and reported data support that the vascular system is an aldone-responsive tissue, and vascular MR has a role in regulating normal vascular function and promoting vascular disease. In vitro data, in vivo animal studies, and human data all indicate a role for MR activation in promoting vascular oxidative stress, inhibiting vascular relaxation, promoting vascular inflammation, fibrosis, and remodeling. These deleterious vascular effects of MR activation appear to be independent of blood pressure changes, with a synergistic effect in the presence of endothelial dysfunction or damage. Thus, in people with underlying cardiovascular disease or cardiovascular risk factors, vascular MR activation may contribute to vascular aging and atherosclerosis, thereby contributing to the pathophysiology of heart attack, stroke, and even hypertension. Further exploring the molecular mechanisms by which MR activates deleterious vascular effects, it would be possible to discover new therapeutic targets to prevent or treat common cardiovascular diseases. Clinical trial data have demonstrated that MR antagonists also have beneficial effects in the treatment of renal diseases, including diabetic nephropathy (International renal journal (Kidney International)2011,79, 1051). The risk of hyperkalemia is currently limiting the use of MR antagonists and particularly excluding diabetic patients. A novel, effective and selective MR antagonist should expand this therapeutic window between endothelial improvement and hyperkalemia of epithelial origin.

WO2006012642 relates to azoles as MR antagonists for the treatment of hypertension, heart failure and similar diseases.

CN103547577 relates to a fused ring compound as an MR antagonist for treating diseases such as renal injury, cardiovascular diseases such as hypertension and the like. US62018790 relates to benzoxazinone amides as MR antagonists for the treatment of cardiovascular and metabolic diseases (e.g. hypertension, heart failure, chronic kidney disease, diabetic nephropathy, diabetes, ocular vasculature diseases).

The 1, 4-dihydro-1, 6-naphthyridine compound (4S) -4- (4-cyano-2-methoxyphenyl) -5-ethoxy-2, 8-dimethyl-1, 4-dihydro-1, 6-naphthyridine-4-deuterium-3-carboxamide is useful as a non-steroidal antagonist of mineralocorticoid receptors and as an agent for the prophylaxis and/or treatment of cardiovascular and renal diseases such as heart failure and diabetic nephropathy.

In organic compounds, many substances often have the same chemical composition and form different crystal structures under different thermodynamic conditions (temperature, pressure, pH, etc.), which we refer to as polymorphism. These different crystalline forms are referred to as "polymorphs". Different polymorphs of a substance have different lattice energies, and thus exhibit different chemical and physical properties in the solid state, including chemical stability, apparent solubility, dissolution rate, melting point, and the like, which can directly affect the handling and production of drug substances and formulations, and can affect the stability, solubility, and bioavailability of drug substances and formulations. It is therefore necessary to prepare more suitable crystalline forms.

In addition, 1, 4-dihydro-1, 6-naphthyridine compound (4S) -4- (4-cyano-2-methoxyphenyl) -5-ethoxy-2, 8-dimethyl-1, 4-dihydro-1, 6-naphthyridine-4-deuterium-3-carboxamide and its preparation method are described in the patent publication CN113214248A, and the synthesis of compound (IX) is performed by using 3-keto-methyl butyrate and 2-cyanoethanol under the activation condition of NBS, the yield is low and the product purification is difficult. The synthesis of the compound deuterated aldehyde (XIVa) is a hydrolysis reaction after the reaction of deuterated DMF and lithiated iodobenzene, has higher synthesis cost and lower reaction temperature, and is not beneficial to large-scale process synthesis. (XIVa) then produces compound (X) by catalytic cyanation of metallic palladium. The compounds (IX) and (X) are condensed under acidic conditions to form a mixture of (VIIa) and (VIIb), and then the mixture and the compound (VIII) undergo a condensation cyclization reaction to obtain the compound (VI). (VI) is reacted with triethyl orthoacetate to give (V), which is hydrolyzed to give (IV), which is then condensed and aminated with aqueous ammonia to give the racemate (R, S) - (I), and this reaction is poor in reproducibility and unstable in yield. Finally, the compound (I) is obtained by SFC chiral resolution. Although SFC resolution can yield compound (I) in high chiral purity, the purchase and production costs and handling of SFC equipment under GMP conditions can present significant challenges. There is also a need to develop synthetic routes that are simple to operate and more cost effective. The specific reaction formula of the original route is as follows:

disclosure of Invention

The invention develops an efficient preparation method of (4S) -4- (4-cyano-2-methoxyphenyl) -5-ethoxy-2, 8-dimethyl-1, 4-dihydro-1, 6-naphthyridine-4-deuterium-3-formamide on the basis of researching the prior art, can provide products with good stability and high chiral purity, and simultaneously researches a crystal form A and a crystal form B to provide support for later formulation research, thereby more effectively treating heart failure and diabetic nephropathy related diseases and meeting different clinical medication requirements.

The invention provides a crystal form A of (4S) -4- (4-cyano-2-methoxyphenyl) -5-ethoxy-2, 8-dimethyl-1, 4-dihydro-1, 6-naphthyridine-4-deuterium-3-formamide shown in a formula (I).

The X-ray diffraction pattern of the crystal form A obtained by the invention has main characteristic peaks with peak maximum values at 2 theta angles of 8.547 degrees, 11.400 degrees, 14.060 degrees, 15.392 degrees, 19.017 degrees, 19.763 degrees, 22.939 degrees, 23.904 degrees, 25.562 degrees and 26.467 degrees.

The crystal form A of the compound shown in the formula I has DSC endothermic transition at 254 +/-2 ℃, has no weight loss in the process of heating to 150 ℃, and can be decomposed at the temperature of more than 255 ℃.

Further, a crystal form TG/DSC spectrum of the compound A shown in the formula I is shown in figure 2.

Furthermore, v of infrared light absorption spectrum of crystal form A of the compound shown in the formula I_maxAt 3475.6, 3417.7, 3364.7, 2990.8, 2952.2, 2939.6, 2229.1, 1682.4, 1657.2, 1607.1, 1574.5, 1483.1, 1432.2, 1324.7, 1169.8, 1031.3, 875.3cm^-1The main characteristic peak with the maximum value of the peak.

The invention also reports a method for preparing the crystal form A of the compound shown in the formula I, which comprises the following steps:

1) mixing (4S) -4- (4-cyano-2-methoxyphenyl) -5-ethoxy-2, 8-dimethyl-1, 4-dihydro-1, 6-naphthyridine-4-deuterium-3-carboxamide with a suitable amount of an inert solvent, stirring at a temperature of 20 ℃ to 120 ℃;

2) dropwise adding a proper amount of benign solvent until the solid is just completely dissolved, cooling the solution to 0-30 ℃, and precipitating to obtain A crystal form;

3) after filtration, the filter cake was dried under vacuum at 35 ℃ and 5 ℃.

Further, the inert solvent comprises isopropyl acetate, n-heptane, cyclohexane, diethyl ether, methyl tert-butyl ether, water, toluene; benign solvents include dimethyl sulfoxide, methanol, acetone, ethylene glycol methyl ether, dimethylformamide, dichloromethane, chloroform, ethanol, n-propanol, isopropanol, 4-methyl-2-pentanone, ethyl acetate, ethyl formate, butyl formate, dioxane, ethylene glycol dimethyl ether, acetonitrile.

The invention also discloses the use of the compound of crystal form A for the manufacture of a medicament for the treatment of cardiovascular disorders and diseases associated with diabetic nephropathy.

The invention also provides a B crystal form of (4S) -4- (4-cyano-2-methoxyphenyl) -5-ethoxy-2, 8-dimethyl-1, 4-dihydro-1, 6-naphthyridine-4-deuterium-3-formamide shown in the formula (I).

The X-ray diffraction pattern of the B crystal form obtained by the invention has main characteristic peaks with peak maximum values at 2 theta angles of 85.792 degrees, 10.273 degrees, 10.635 degrees, 11.267 degrees, 12.048 degrees, 14.834 degrees, 21.849 degrees, 22.535 degrees, 24.171 degrees and 24.734 degrees.

The crystal form B of the compound shown in the formula I is characterized in that DSC endothermic transition is 120 +/-2 ℃ and 252 +/-2 ℃, 12.3 percent of weight loss is generated in the process of heating to 120 ℃, and decomposition is possible at the temperature of above 255 ℃.

The invention also discloses a method for preparing the B crystal form of the compound shown in the formula I, which comprises the following steps:

1) mixing (4S) -4- (4-cyano-2-methoxyphenyl) -5-ethoxy-2, 8-dimethyl-1, 4-dihydro-1, 6-naphthyridine-4-deuterium-3-carboxamide with a proper amount of inert solvent, and stirring at the temperature of 20-120 ℃;

2) dropwise adding a proper amount of benign solvent until the solid is just completely dissolved, cooling the solution to 0-30 ℃, and precipitating a crystal form A;

3) after filtration, the filter cake was dried under vacuum at 35 ℃ and 5 ℃.

Further, the inert solvent comprises isopropyl acetate, n-heptane, cyclohexane, diethyl ether, methyl tert-butyl ether, water, toluene; the benign solvent is tetrahydrofuran.

Further, the cooling temperature in the above step 2 is preferably 0 to 5 ℃.

The invention also discloses the use of the compound of crystal form B for the manufacture of a medicament for the treatment of cardiovascular disorders and diabetic nephropathy-associated diseases.

Compounds of formula (I) and chirality thereofThe purification preparation method is described in the patent publication CN113214248A, and comprises performing supercritical fluid chromatography with Waters UPCC (CA-119) of SFC, and subjecting to DAICEL column chromatography

AS-3(100 x 3mm 3 μm) and mobile phase supercritical carbon dioxide and methanol (0.1% diethylamine). Although SFC separation can achieve relatively good yields and optical purity, the cost of acquisition and operation of such equipment under GMP conditions is difficult. And simultaneously, the feasibility of producing products in kilogram level or ton level is extremely low along with high cost.

In view of the above problems, the present invention also provides an improved process for the synthesis of compounds (IX), (X) and (R, S) - (I) which is convenient for large scale operations, wherein the preparation of high purity chiral compound I by an alternative to the separation of enantiomeric mixtures can be carried out using conventional pilot plant equipment (stirred vessel/separation unit) and does not require the purchase of additional special equipment, overcoming the disadvantages of low yield and high cost in the original published synthetic routes, difficult purification and not easy to produce on a large scale.

In order to solve the technical problems, the synthesis method for preparing the compound of the formula (I) provided by the invention comprises the following steps:

the synthesis method comprises the following steps:

1. compound XI is prepared by a boronic acid catalyzed transesterification reaction to compound IX;

2. the starting material is subjected to a catalytic cyanation reaction of compound XIII palladium metal to obtain compound XIV;

3. preparing compound X from deuterated compound XIV under the action of compound XVI;

4. carrying out Knoevenagel dehydration condensation reaction on the compound IX and the compound X to obtain a mixture of a compound VIIa and a compound VIIb;

5. preparing a compound VI by a condensation cyclization reaction of a mixture of a compound VIIa and a compound VIIb and a compound VIII;

6. carrying out etherification reaction on the compound VI to obtain a compound V;

7. preparing a compound IV from the compound V through hydrolysis reaction;

8. the compound IV firstly generates carbonyl imidazole under the action of a condensing agent CDI, and then the carbonyl imidazole is subjected to amidation reaction with HMDS and then is hydrolyzed by a one-pot method to prepare a compound (R, S) -I;

9. the compound (R, S) -I reacts with chiral tartaric acid dibenzoate to obtain a compound XVII;

10. compound XVII is liberated by addition of base to give compound I.

Specifically, in step 1, compound XI and compound XII are reacted in toluene with boric acid at an ambient temperature of 110 ℃ to provide compound IX. More specifically, compound (IX) is an important starting material, and compound IX is obtained by reacting compound (XI) and (XII) in toluene (5-8 times by volume) with the addition of 10-20 mol% boric acid at 110 ℃ C (external temperature, internal temperature 100 ℃ C.) for 12-15 hours. The reaction utilizes ester exchange catalyzed by boric acid, and has the advantages of few byproducts, high yield, easy purification of products and easy amplification.

Specifically, in step 2, compound XIII, potassium hexachloroferricyanide (20-30 mol%) and sodium carbonate (1 mol%) are reacted in N, N-dimethylformamide (5-8 times by volume) with palladium acetate (0.5 mol%) at 120 ℃ under nitrogen atmosphere for 12-15 hours to obtain compound XIV.

Specifically, in step 3, compound XV is first reacted with triethyl orthoformate (2.5 to 3 equivalents) in a mixed solution of acetic acid (2.5 times by volume) and water (0.5 times by volume) at room temperature for 12 to 15 hours, and water (10 times by volume) and ammonium tetrafluoroborate (1.5 equivalents) are added and stirred at room temperature for 30 to 50 minutes to obtain compound XVI, as shown in the following formula:

compound XVI (2-4 mol%) and compound XIV (1 equivalent) are then reacted in toluene (10 volumes) and deuterated deuterium oxide (10 volumes) with sodium bicarbonate (1 equivalent) under nitrogen at 100 deg.C (external temperature, internal temperature 85-90 deg.C) for 5-8 hours to give compound X.

The compound X is an important intermediate of the compound I, and the compound (XIV) is subjected to deuteration by reaction with commercially available heavy water which is easy to start under the action of the carbene catalyst (XVI), so that the compound X and the synthesis cost are greatly reduced. The carbene catalyst (XVI) can be obtained by reacting commercially available raw material (XV) and triethyl orthoformate (2.5-3 equivalents) in a mixed solution of acetic acid (2.5 times volume) and water (0.5 time volume) at room temperature for 12-15 hours, and adding water (10 times volume) and ammonium tetrafluoroborate (1.5 equivalents) for reaction.

Specifically, in step 4, compound IX and compound X are reacted in isopropanol (8-10 volumes) with acetic acid (10-20 mol%) and piperidine (10-20 mol%) for 2-3h at room temperature to give a mixture of compound VIIa and compound VIIb.

Specifically, in step 5, a mixture of compound VIIa and compound VIIb is reacted with compound VIII in an ethanol solution (8-10 times by volume) at 95 ℃ (external temperature, internal temperature 78-82 ℃) to give compound VI.

Specifically, in step 6, compound VI and triethyl orthoacetate (3-5 equiv.) are reacted in NMP with concentrated sulfuric acid (0.5-0.6 equiv.) at 120 deg.C (outer temperature, inner temperature 105-.

Specifically, in step 7, the volume ratio of compound V to aqueous sodium hydroxide solution in THF/water is 2:1 at 0c to give compound IV.

Specifically, step 8 is a one-pot procedure in which compound IV is first reacted with diimidazole carbonate and a catalytic amount of 4- (dimethylamino) pyridine in THF to produce a carbonylimidazole intermediate; then adding hexamethyldisilane into the reaction liquid to generate N-silane amide; finally hydrolyzing to obtain the compound (R, S) -I.

More specifically, the original preparation method of (R, S) - (I) utilizes the reaction of compound (IV) with ammonia under the action of a condensing agent HATU, which is not easy to operate on a large scale and has poor reproducibility and unstable yield, and the improved method provided by the invention is that compound (IV) is reacted with (1.3-1.4 equivalent) 1,1' -dicarbonyl imidazole in THF for 2-3 hours at 50-60 ℃ in the presence of catalytic amount of 4- (dimethylamino) pyridine (0.1-0.2 equivalent); then hexamethyldisilazane (3-4 equiv) is directly added to continue reacting for 3-4 hours under 85-90 ℃ (external temperature, internal temperature 70-75 ℃); finally, a mixed solution of THF and water is added dropwise to perform hydrolysis reaction for 1-2 hours at 85-90 ℃ (external temperature, internal temperature of 70-75 ℃), and (R, S) - (I) become white solid to precipitate, and finally, the mixture is filtered, washed by THF and water and dried to obtain the compound (R, S) - (I) with higher purity.

Specifically, in step 9 and step 10, compound (R, S) -I is resolved into compound I and/or compound Ia by using compound IIIa or IIIb.

More specifically, the invention is achieved by using chirally substituted dibenzotartrate of formula (IIIa)

Resolution of the racemic (R, S) - (II) compound, reaction in an alcohol/water mixture to give the diastereomeric salt of formula (XVII), wherein the enantiomer with S configuration preferably forms a salt. Almost quantitatively, the diastereomeric salt (XVII) precipitates out of solution, which can then be separated from the solution, for example by filtration, while the enantiomer with R configuration remains in solution.

Alternatively, in a very similar manner, the mirror salt of formula (XVIIa) is prepared by reacting the racemate (R, S) - (II) with a tartaric acid derivative of formula (IIIb), wherein the enantiomer with the R configuration preferably forms a salt. The precipitated diastereomeric salt can be separated off almost quantitatively and the S-enantiomer remains in solution here.

Tartaric acid derivatives of compound I having S configuration, S-configuration are preferably used for racemate resolution, which reacts in combination with racemates (R, S) - (II) preferentially to form diastereomeric salts of the S-enantiomer.

More specifically, the chirally substituted dibenzoate is particularly preferably 0.50 to 0.55 equivalent for resolution of the racemate (R, S) - (II).

More specifically, the formation of the diastereomeric salt of formula (XVII) and/or formula (XVIIa) is carried out in a solvent mixture consisting of water and a water-miscible organic solvent.

More specifically, suitable organic solvents used for the formation of diastereomeric salts are, for example, ethanol, methanol, isopropanol, 1-propanol, isobutanol, 1-pentanol or acetone; however, ethanol is preferably used. The solvents can also be used in denatured form, which is commercially available, for example denaturants which are customary for ethanol, such as toluene, hexane, heptane.

More specifically, the diastereomeric salt of formula (XVII) and/or formula (XVIIa) can be isolated by filtration, followed by separation and purification to the corresponding highly pure chiral compound of formula (I) or formula (Ia) by addition of a base to treat the diastereomeric salt with water in a solvent mixture of water and a water-miscible organic solvent and removing the solvent.

More specifically, the base used to treat the diastereomeric salt is an inorganic base. Preferred bases are aqueous sodium hydroxide solution, lithium hydroxide, potassium hydroxide, ammonium carbonate, sodium carbonate, potassium carbonate, lithium carbonate, ammonium bicarbonate, sodium bicarbonate, potassium bicarbonate, sodium phosphate, potassium phosphate, ammonium phosphate. Sodium or potassium phosphate is particularly preferred.

In other aspects, the invention also provides a pharmaceutical composition comprising a pharmaceutically acceptable carrier and a compound of the invention; methods of treating or preventing disorders or conditions selected from utility in cardiovascular and metabolic diseases, clinical disorders such as hypertension, heart failure, chronic kidney disease, diabetic nephropathy, fibrosis, endothelial dysfunction and muscle atrophy, and to methods for their potential therapeutic use.

The compound (4S) -4- (4-cyano-2-methoxyphenyl) -5-ethoxy-2, 8-dimethyl-1, 4-dihydro-1, 6-naphthyridine-4-deuterium-3-carboxamide of formula (I) provided in the present invention and its specific form a and form B may be used in combination with one or more additional agents independently selected from the group consisting of: cardiac therapeutics, antihypertensives, diuretics, peripheral vasodilators, lipid regulators, antidiabetics, anti-inflammatory agents, or anticoagulants.

The above cardiotherapeutic, antihypertensive, diuretic, peripheral vasodilator, lipid regulating, antidiabetic, antiinflammatory, or anticoagulant agents include, but are not limited to, digitoxin, antiarrhythmic agents (e.g., amiodarone, brombenium, amiodarone), calcium channel antagonists (e.g., amlodipine, felodipine, nifedipine sustained release tablets, nifedipine controlled release tablets, lacidipine, etc.), ACE inhibitors (e.g., captopril, enalapril, ramipril, perindopril, benazepril, fosinopril, etc.), angiotensin II receptor blockers (e.g., losartan, valsartan, irbesartan, telmisartan, olmesartan, candesartan, olmesartan), endothelin receptor blockers, beta-blockers (e.g., propranolol, metoprolol, propranolol, etc.), and, Thiazide diuretics (e.g., chlorothiazide, hydrochlorothiazide (hydrochlorothiazide), cyclopenthiazide, benfluthiazide, chlorothiadone, and the like), loop diuretics (e.g., furosemide, ethacrynic acid, bumetanide, piretanide, torasemide, and the like), cholesterol synthesis inhibitors such as statins (e.g., rosuvastatin, cholesterol absorption inhibitors, Cholesterol Ester Transfer Protein (CETP) inhibitors, antidiabetic agents such as insulin and analogs, GLP-1 analogs, sulfonamides, dipeptidyl peptidase 4(DPP4) inhibitors, thiazoldiones, SGLT-2 inhibitors, metformin, and anti-inflammatory agents such as NSAID and CCR2 antagonists, low molecular weight heparin calcium, enoxaparin, naratriptan, low molecular weight heparin sodium, dalteparin sodium, thrombin inhibitors, and factor Xa inhibitors (e.g., rivaroxaban, apixaban, dabigatran), and platelet aggregation inhibitors (e.g., ticagrelor, Eptifibatide, aspirin, dipyridamole, clopidogrel).

Pharmaceutical compositions containing compound I of the present invention or its particular forms a and B are useful for the treatment and/or prevention of diseases or disorders mediated by or affected by one or more steroid nuclear receptors, or associated with steroid nuclear receptor activity, such as clinical disorders like hypertension, heart failure, chronic kidney disease, diabetic nephropathy, fibrosis, endothelial dysfunction and muscle atrophy.

Pure forms or suitable pharmaceutical compositions of compound I of the present invention, its forms a and B, its pharmaceutically acceptable salts, may be administered by any acceptable mode of administration of agents that serve similar utilities. The pharmaceutical compositions of the invention may be prepared by combining a compound of the invention with a suitable pharmaceutically acceptable carrier, diluent or excipient, and may be formulated into solid, semi-solid, liquid or gaseous form preparations, such as tablets, capsules, powders, granules, solutions, injections, inhalants, gels, microspheres and aerosols. Typical routes of administration of the pharmaceutical composition include, but are not limited to, oral, topical, transdermal, inhalation, parenteral, sublingual, buccal, rectal, and intranasal administration. As used herein, the term parenteral includes subcutaneous injections, intravenous, intramuscular, intrasternal injection or infusion techniques. The pharmaceutical compositions of the present invention are formulated to allow the active ingredients contained therein to be bioavailable upon administration of the composition to a patient. One or more dosage units in the composition to be administered to a subject or patient, wherein, for example, a tablet may be a single dosage unit and a container containing a compound of the invention in aerosol form may contain a plurality of dosage units. The actual methods of preparing the dosage forms are known, or will be known, to those skilled in the art. The compositions to be administered will contain, in any event, a therapeutically effective amount of a compound of the present invention, or a pharmaceutically acceptable salt thereof, in order to treat the disease or condition of interest in accordance with the teachings of the present invention.

The pharmaceutical compositions of the present invention may be in solid or liquid form. In one aspect, the carrier is a microparticle, such that the composition is in the form of, for example, a tablet or powder. The carrier can be a liquid and the composition can be, for example, an oral syrup, an injectable liquid, or an aerosol suitable, for example, for administration by inhalation. When intended for oral administration, the pharmaceutical composition is preferably selected from solid or liquid forms, wherein semi-solid, semi-liquid, suspension and gel forms are included in the forms considered herein as solid or liquid. For oral solid compositions, the pharmaceutical compositions may be formulated into powders, granules, compressed tablets, pills, capsules, chewable tablets, powder tablets, and the like. Such solid compositions typically contain one or more inert diluents or edible carriers. In addition, one or more binders such as carboxymethyl cellulose, ethyl cellulose, microcrystalline cellulose, xanthan gum or gelatin may also be present; excipients, such as starch, lactose or dextrin; disintegrating agents, such as alginic acid, sodium alginate, Primogel, corn starch, and the like; lubricants, such as magnesium stearate or hydrogenated vegetable oil (Sterotex); glidants, such as colloidal silicon dioxide; sweetening agents, such as sucrose or saccharin; flavoring agents, such as peppermint, methyl salicylate or orange flavoring; and a colorant.

Where the pharmaceutical composition is in the form of a capsule, for example a gelatin capsule, it may contain, in addition to materials of the above type, a liquid carrier such as polyethylene glycol or a vegetable oil. The pharmaceutical composition may be in liquid form, such as a tincture, syrup, solution, emulsion or suspension. Such liquids may be administered orally, or delivered by injection, as two examples. When intended for oral administration, the compositions preferably contain one or more of sweeteners, preservatives, dyes/colorants and flavor enhancers in addition to the compounds of the present invention. In compositions intended for administration by injection, one or more of surfactants, preservatives, wetting agents, dispersing agents, suspending agents, buffering agents, stabilizing agents and isotonicity agents may be included. Oral and parenteral administration, especially oral administration, are preferred.

Drawings

FIG. 1: the XRPD spectrogram of the crystal form A compound of the invention;

FIG. 2: TG/DSC spectrogram of the crystal form A compound of the invention;

FIG. 3: the invention relates to an infrared spectrogram of a crystal form A compound;

FIG. 4: an XRPD spectrogram of the B crystal form compound of the invention;

FIG. 5: TG/DSC spectrogram of the B crystal compound;

FIG. 6: compound (I) single crystal molecular ball-stick model;

FIG. 7: compound (I) cell ball and stick model;

FIG. 8 is an optical microscope image of the compound of crystal form A of the present invention under (a) a polarizer and (b) no polarizer;

FIG. 9: the compound A crystal form of the invention is used for carrying out drug effect experiment on animal urine Na in a deoxycorticosterone acetate/sodium chloride load-induced WKY rat nephropathy model⁺，K⁺(ii) a change;

FIG. 10: the evaluation results of hematoxylin eosin staining (HE) glomerulosclerosis and renal tubule pathological tissue sections in a WKY rat nephrosis model pharmacodynamic experiment induced by the compound A crystal form deoxycorticosterone acetate/sodium chloride load are shown.

Detailed Description

The invention is further illustrated by the following examples, which are provided to illustrate the compounds, pharmaceutical compositions, and methods provided herein and by way of illustration of the synthetic examples and biological examples described herein. The following examples are intended to illustrate the present invention, but are not intended to limit the present invention, and modifications, changes, variations, etc. within the scope of the present invention are included.

The compounds provided herein can be prepared from readily available starting materials using the procedures of the particular synthetic schemes set forth below, which will be well known to those skilled in the art. Experimental procedures, under which specific conditions are not noted in the following examples, can be determined by those skilled in the art through routine optimization procedures, according to conventional methods and conditions.

In the following examples, abbreviations are explained:

PE: petroleum ether;

EA: ethyl acetate;

DMF: n, N-dimethylformamide;

DCM: dichloromethane;

THF: tetrahydrofuran (THF)

MeOH: methanol

K₄Fe(CN)₆Potassium hexachloroferricyanide

Pd(OAc)₂Palladium acetate

HMDS hexamethyldisilazane

CDI: 1,1' -carbonyldiimidazole

DMAP: 4- (dimethylamino) pyridine

CDCl₃: deuterated chloroform DEA: diethylamine

Piperidine: piperidine derivatives

IPA: isopropanol (I-propanol)

Na₃PO₄: sodium phosphate

SFC: supercritical fluid chromatography

MS: mass spectrometry

ESI: electrospray ionization

¹H NMR: hydrogen spectrum of nuclear magnetic resonance

TLC: thin layer chromatography

Chiral HPLC: chiral high performance liquid chromatography

Prep-HPLC high pressure preparative liquid chromatography

LC-MS: liquid chromatography-mass spectrometry combination

Rf: a ratio shift value;

min: minute (min)

g; keke (Chinese character of 'Keke')

mg: milligrams of

rt: at room temperature

mol: mole of

mmol: millimole

mL: milliliter (ml)

M: mole/liter

Example 1: (S) -4- (4-cyano-2-methoxyphenyl) -5-ethoxy-2, 8-dimethyl-1, 4-dihydro-1, 6- Preparation of naphthyridine-4-deuterium-3-carboxamide (Compound I):

chemical combination ofPreparation of substance (XVI): compound (XV) (30.0g,78.9mmol), triethyl orthoformate (29.1g,196.6mmol) were dissolved in a mixed solution of acetic acid (78mL) and water (18mL), and the reaction was carried out at room temperature for 12 hours. LCMS monitored the reaction with 70% product formation and 30% starting material. After the reaction was completed, the reaction solution was added to water (300mL), ammonium tetrafluoroborate (9.6g,109mmol) was added, stirred at room temperature for 30 minutes, filtered, and the filter cake was rinsed with water (300mL X4) to obtain 15g of filter cake. The mother liquor was extracted with dichloromethane (200mL X3), the organic phases were combined and dried over anhydrous sodium sulfate, concentrated under reduced pressure and dried to give the crude product (10 g). The combined crude products were slurried with methanol, petroleum ether (PE: EA ═ 5:1) (500 mL). Clean compound (XVI) (15g, 48.6% yield) was obtained as a white solid. MS (ESI) M/z 391.32[ M ]]⁺。

Preparation of compound (IX): compound (XI) (2500g, 21.55mol), compound (XII) (1530g, 21.55mol) and boric acid (267.2g, 4.31mol) were dissolved in 15L of toluene and reacted at 110 ℃ C (external temperature, internal temperature 100 ℃ C.) for 12 hours. TLC monitored HNN4-1 remaining but product was complete. After the reaction is finished, cooling the reaction system to 40-60 ℃, extracting the water + phase by using ethyl acetate (2L X2), combining the organic phases, and drying by using anhydrous sodium sulfate. Concentrating under reduced pressure and spin-drying to obtain crude product. Column chromatography (PE: EA ═ 5: 1-2: 1) of the crude product afforded compound (IX) (2100g, 62% yield) as a white solid. MS (ESI) M/z 215.97[ M +1] +.

Preparation of compound (XIV): compound (XIII) (500g,2325mmol), potassium hexacyanoferrate (222.7g,604.5mmol) and sodium carbonate (2325mmol) were charged into a 10L four-necked reaction flask, 4L of DMF (25L in a barrel) was charged into the reaction flask, stirred at room temperature, replaced with nitrogen three times, followed by addition of palladium acetate (2.61g, 11.63mmol) to the reaction mixture, replaced with nitrogen three times, and the reaction was carried out at 120 ℃ under nitrogen for 12 hours. The completion of the reaction was monitored by TLC (PE: EA ═ 2: 1). After the three batches of combined treatment reactions are completed, cooling the reaction system to 40-60 ℃, adding the reaction solution into water (12L), stirring for 30 minutes at room temperature, filtering by using kieselguhr, and leaching the filter cake by using water (2L X4). The filter cake was added to ethyl acetate (20L) and stirred at room temperature for 30 minutes, the stirred cake was filtered, rinsed with ethyl acetate (2L X2), the organic phases were combined and dried with waterAfter drying over sodium sulfate, concentration under reduced pressure and spin-drying, the crude product (900g) was obtained. The filtrates were collected separately, and the filtrate (20L) was extracted with ethyl acetate (10L X3), washed with saturated brine (5L), the organic phases were combined and dried over anhydrous sodium sulfate, concentrated under reduced pressure and dried by rotary drying to give the crude product (450g, crude). The combined crude products were slurried with methanol, petroleum ether (MeOH: PE ═ 1:2) (1.5L). Clean product (750g) was obtained. Column chromatography of the mother liquor (PE: EA: 5:1) gave a further clean product (225 g). In total, compound (XIV) (975g, yield 87%) was obtained as a white solid. MS (ESI) M/z 163.05[ M +1]]+；¹H NMR(400MHz,DMSO-d6)δ3.98(s,3H),7.52-7.54(m,1H),7.79-7.83(m,2 H)，9.58(s,1H)。

Preparation of Compound (X): compound (XIV) (550g, 3416mmol) and sodium hydrogencarbonate (287g, 3416mmol) were charged into a 20L four-necked reaction flask, and a mixed solution of heavy water (5500mL) and toluene (5500mL) was charged into the reaction flask, and stirred at room temperature, and nitrogen was substituted three times, and then compound (XVI) (53.4g, 136.6mmol) was added to the reaction mixture, and nitrogen was substituted three times, and the reaction was carried out at 100 ℃ C (outer temperature, inner temperature 85-90 ℃ C.) under nitrogen atmosphere for 5 hours. The HNMR monitored the reaction to completion. After the reaction is finished, cooling the reaction system to 40-60 ℃, separating the reaction liquid, extracting the heavy water phase with dichloromethane (2L X2), combining the organic phases and drying with anhydrous sodium sulfate. The organic phase was concentrated under reduced pressure until 1.5L of toluene remained, and petroleum ether (3L) was added to the toluene phase. The temperature is reduced to-60 ℃, and the solution is stirred for 30 minutes at-60 ℃. Filtration and drying of the filter cake gave the product (400 g). Column chromatography of the mother liquor (PE: EA ═ 2:1) afforded a clean product (30g) which amounted to a solid product (430g, 78%). MS (ESI) M/z 163.05[ M +1] +.

Preparation of compounds (VIIa) and (VIIb): compound (X) (200g, 1235mmol), piperidine (20.7g, 246.9mmol) and acetic acid (14.8g, 246.9mmol) were added to a 5L four-necked reaction flask, followed by addition of a solution of isopropanol (1200mL), stirring at room temperature, addition of a solution of compound (IX) (195.2g, 1259.3mmol) in isopropanol (400mL) to the reaction flask, and reaction at room temperature for 2 hours. The completion of the reaction was monitored by TLC (PE: EA ═ 2: 1). After the reaction was completed, the reaction system was cooled to-60 ℃ with a dry ice ethanol bath, and petroleum ether (2L) was added to the reaction solution to react at-60 ℃ with stirring for 30 minutes. Filtering under reduced pressure, eluting the reaction solution with petroleum ether (1L), cold isopropanol (1L) and petroleum ether (500mL X3), and drying the filter cake to obtain a mixture of compounds (VIIa) and (VIIb) as a yellow solid (320g, 87% yield). MS (ESI) M/z 300.10[ M +1] +.

Preparation of Compound (VI): adding a mixture (600g, 2007mmol) of the compounds (VIIa) and (VIIb) into a stirring solution of isopropanol (50mL), adding the compound (VIII) 4-amino-5-methylpyridin-2-ol (248.9g, 2007mmol) into a 10L four-mouth reaction bottle, adding an ethanol (6L) solution into the reaction bottle, stirring at room temperature, protecting with nitrogen, and reacting at 95 ℃ (external temperature, internal temperature of 78-82 ℃) under nitrogen atmosphere for 12 hours. After the reaction is completely finished by LCMS and TLC monitoring, cooling the reaction system to 40-60 ℃, cooling the reaction system to-60 ℃ by using a dry ice ethanol bath, adding petroleum ether (5L) into the reaction solution, and stirring for 30 minutes at-60 ℃ for reaction. The reaction mixture was filtered under reduced pressure, the reaction mixture was rinsed with cold ethanol (1L X2) and petroleum ether (2L X2), respectively, and the filter cake was dried to give the product (500 g). The mother liquor was concentrated under reduced pressure and spin-dried, and purified by column chromatography to give the product (120g), which was combined to give the solid compound (VI) as a white solid (620g, yield 76%). LCMS M/z 406.1[ M + H ]]⁺。

Preparation of Compound (V): compound (VI) (300g, 740.7mmol), triethyl orthoacetate (600g, 3704mmol) were charged to a 5L four-necked reaction flask, NMP (1500mL) solution was added to the flask, stirred at room temperature under nitrogen, followed by H₂SO₄(45mL) the reaction was carried out at 120 deg.C (external temperature, internal temperature 105 ℃ C.) and 110 deg.C for 2 hours. LCMS and TLC monitor the reaction is complete. After the reaction is finished, cooling the reaction system to 40-60 ℃, and adding Na into the reaction liquid at 0 DEG C₂CO₃(135.0g) was added to the saturated solution, and the reaction was stirred at 0 ℃ for 30 minutes. The reaction mixture was filtered under reduced pressure, rinsed with cold water (500mLX 2) and petroleum ether (500mL X2), respectively, the filter cake was dried to give a crude product (255g), which was slurried with methanol to give pure compound (V) (220 g). The mother liquor was extracted with ethyl acetate (3L X2), the organic phase was washed with saturated brine (500mL), dried over anhydrous sodium sulfate, concentrated under reduced pressure and spin-dried, and the crude product was slurried with methanol to give the pure product (30.0 g). Are combined to give compound (V)White solid (250g, yield 78%). LCMS M/z 381.1[ M + H ]]⁺。

Preparation of Compound (IV): compound (V) (200g, 462mmol) was charged into a 5L four-necked reaction flask, a mixed solution of THF (1200mL) and water (600mL) was added to the reaction flask, and the reaction was allowed to cool to 0 ℃. A solution of NaOH (24.0g,600.6mmol) in water (400mL) was added dropwise at 0 ℃ for 30 minutes, and the reaction was carried out at 0 ℃ for 1.5 hours. LCMS, TLC monitored reaction complete. The stirring was stopped and the reaction solution was extracted with methyl tert-butyl ether (800mL X2) and ethyl acetate (800 mL). The pH was adjusted to 7 with dilute hydrochloric acid at 0 ℃ and 200g NH were added₄Cl and 800mL of water. After the solid was precipitated, suction filtration was carried out under reduced pressure, the reaction solution was rinsed with water (500mL X2) and petroleum ether (500mL X2), and the filter cake was dried to obtain compound (IV) (150g, 91.0% yield) as a white solid. LCMS M/z 380.2[ M + H ]]⁺。

Preparation of Compound (R, S) - (I): compound (IV) (150g, 395mmol), CDI (96.0g, 592mmol) and DMAP (4.82g, 39.5 mmol) were charged into a 2L three-necked reaction flask, a THF (750mL) solution was added to the reaction flask at room temperature, nitrogen was substituted three times, and the reaction was allowed to react at room temperature for 1 hour. The temperature is raised to 50 ℃ (external temperature, internal temperature 40-45 ℃), and the reaction is carried out for 2 hours at 50 ℃. TLC (PE: EA ═ 1:1) showed complete reaction of starting material. HMDS (318.8g, 1975mmol) was then added to the reaction, and the reaction was allowed to react at 85 deg.C (outer temperature, inner temperature 70-75 deg.C) for 3 hours. LCMS and TLC showed the starting material reaction was complete. The reaction was allowed to fall to 0 ℃ and a mixed solution of THF (300mL) and water (150mL) was added dropwise at 0 ℃. After the completion of the dropwise addition, the reaction solution was heated to 85 ℃ (external temperature, internal temperature 70-75 ℃), reacted at 85 ℃ (external temperature, internal temperature 70-75 ℃) for 1 hour, and then cooled to 0 ℃. LCMS, TLC monitored reaction complete. Filtration under reduced pressure, rinsing the reaction solution with water (300mL X2), drying the filter cake in a forced air oven and a vacuum oven to obtain the compound (R, S) - (I) (120g, 80.2%) as a white solid. LCMS M/z 380.2[ M + H ]]⁺。

Preparation of compound (XVII): (R, S) - (I) (120g, 316.6mmol) was charged into a 5L four-necked flask, and a mixed solution of ethanol and water (1980mL) was added to the flask, followed by addition of Compound (IIIa) D + tartrate benzoate (62.3g, 174.1 mmol)). The reaction was stirred at room temperature for 30 minutes, then warmed to 90 ℃ at an internal temperature of 75 ℃ and reacted at an internal temperature of 75 ℃ for 3 hours. The reaction was allowed to cool to room temperature over 5 hours. The reaction was then allowed to react at room temperature for 12 hours. SFC monitored reaction completion. The reaction solution was filtered, washed with ethanol: the reaction was washed with 3:1(280mL X2) and the filter cake was dried in a forced air oven for 2 hours. Compound (XVII) (100g, 43%) was obtained as a white solid. LCMS M/z 380.2[ M + H ]]⁺。¹H-NMR(400MHz，DMSO-d₆)： δ＝1.06(t,3H)，2.13(s,3H)，2.18(s,3H)，3.83(s,3H)，3.99-4.07(m,2H)，5,89(s,2H)， 6.61-6.85(m,2H)，7.15(d,1H)，7.28(dd,1H)，7.38(d,1H)，7.56(s,1H)，7,62(t,4H)，7.69(s, 1H)，7,76(t,2H)，8,06(d,4H)，12.60-15.430(br s,2H)。

Preparation of Compound (I): tartrate salt (XVII) (100g, 135.7mmol) was charged into a 5L four-necked reaction flask, and a mixed solution (1000mL) of ethanol (200mL) and water (800mL) was added to the reaction flask, followed by dropwise addition of anhydrous Na over 20 minutes₃PO₄(33.4g, 203.5mmol) in water (400 mL). The temperature of the reaction was raised to 65 ℃ C (inner temperature 50 ℃ C., based on the inner temperature), and the reaction was carried out at an inner temperature of 50 ℃ C. for 4 hours. The reaction was allowed to cool to room temperature over 2 hours. SFC monitored reaction completion. The reaction was filtered, rinsed with 1:4(400mL X2) ethanol and the filter cake was dried in a forced air oven for 12 hours. Compound (I) (45g, 87.5%) was obtained as a white solid. LCMS M/z 383.2[ M + H ]]⁺；¹H NMR(400MHz,DMSO-d6)δ1.03-1.07(m,3 H),2.19(s,3H),3.82(s,3H),4.00-4.02(m,2H)，6.67-6.74(m,2H),7.15(d,J＝8Hz,1H), 7.26-7.29(m,1H),7.37(s,1H),7.55(s,1H),7.69(s,1H)。

Table 1 HPLC analysis conditions:

the purity of the compound I is 99.27%;

table 2 SFC analysis conditions:

the chiral purity of the (S) form of the compound (I) was 99.4%, and the optical purity was 98.8% ee.

Example 2: (S) -4- (4-cyano-2-methoxyphenyl) -5-ethoxy-2-methyl-8-trideuteromethyl-1, 4- Preparation of crystalline form a of dihydro-1, 6-naphthyridine-4-hydro-3-carboxamide (I):

in a 1000mL single-neck flask, a 40g sample of compound (I) was dissolved in 1.6L ethanol, 6.40L water was added, and the mixture was heated to an internal temperature of 50 ℃ over 1 hour and stirred at that temperature for 4 hours. The mixture was cooled to 22 ℃ over 1 hour and stirred at this temperature for 20 hours, filtered by standing and the filter cake was dried in a vacuum oven (40 ℃) for 20 hours to give 36.5g of an off-white solid.

Preparation of binary solvent crystal form A

In a 100mL single-neck flask, 4g of a sample of compound (I) was dissolved in 100mL of ethanol, 900mL of methyl t-butyl ether was added, and the mixture was stirred at room temperature for 72 hours, allowed to stand and filtered, and the filter cake was dried in a vacuum oven (40 ℃ C.) for 20 hours to give 3.1g of an off-white solid.

TABLE 3 summary of preparation conditions for binary solvent form A

Preparation of single solvent crystal form A

In a 100mL single neck flask, 4g of compound (I) sample was dissolved in 100mL of ethanol, concentrated under reduced pressure to a quarter volume and stirred at room temperature for 20 hours, filtered by standing, and the filter cake was dried in a vacuum oven (40 ℃ C.) for 20 hours to give 3.5g of an off-white solid.

TABLE 4 summary of preparation conditions for single solvent form A

Sample size (mg)	Solvent(s)	Volume (mL)	Temperature (. degree.C.)	Results
					19.8	Ethanol	0.8	50→RT	Crystal form A
19.5	N-propanol	1.0	50→RT→0	Crystal form A
					19.9	Isopropanol (I-propanol)	1.8	50→RT→0	Crystal form A
19.6	Ethyl acetate	1.8	50→RT→0	Crystal form A
					20.1	Acetonitrile	1.0	50→RT	Crystal form A
19.9	Dioxane (dioxane)	0.6	50→RT→0	Crystal form A

The infrared absorption spectrum of the crystal form A prepared by the method is shown in figure 3, v is_maxAt 3475.6, 3417.7, 3364.7, 2990.8, 2952.2, 2939.6, 2229.1, 1682.4, 1657.2, 1607.1, 1574.5, 1483.1, 1432.2, 1324.7, 1169.8, 1031.3, 875.3cm^-1The main characteristic peak with the maximum value of the peak.

Example 3: (S) -4- (4-cyano-2-methoxyphenyl) -5-ethoxy-2-methyl-8-trideuteromethyl-1, 4- Preparation of crystalline form B of dihydro-1, 6-naphthyridine-4-hydro-3-carboxamide (I)

In a 1000mL single neck flask, 2g of compound (I) was dissolved in 200mL tetrahydrofuran, concentrated under reduced pressure to a full volume and stirred at room temperature for 20 hours, filtered by standing, and the filter cake was dried in a vacuum oven (40 ℃ C.) for 20 hours to give 1.5g of an off-white solid.

The preparation of the binary solvent crystal form B comprises the following steps:

s1, mixing a compound I with an inert solvent, and stirring at the temperature of 20-120 ℃;

s2, dropwise adding a benign solvent until the solid is just completely dissolved, cooling the solution to 0-30 ℃, and precipitating a B crystal form;

s3, after filtration, the filter cake is dried in vacuum at 35 +/-5 ℃.

The inert solvent comprises isopropyl acetate, n-heptane, cyclohexane, diethyl ether, methyl tert-butyl ether, water and toluene; the benign solvent is tetrahydrofuran. Preferably, the cooling temperature of the solution of step S2 is 0 ℃ to 5 ℃.

Example 4: experiment of influence factors of moisture absorption

Dynamic water sorption desorption analysis was determined using DVS Intrinsic (SMS, UK). The test adopts a gradient mode, the humidity change is 50% -95% -0% -50%, the humidity change of each gradient in the range of 0% to 90% is 10%, the gradient end point is judged in a dm/dt mode, and the dm/dt is less than 0.002% and is maintained for 10min as the gradient end point. After the test is finished, XRPD analysis is carried out on the crystal form of the sample to confirm whether the solid form is changed. The moisture absorption weight of the crystal form A is increased by 1.73% under 95% humidity, the weight loss of the crystal form A is 0.09% under 0% humidity, and the moisture absorption weight of the crystal form A is increased by 0.40% under 80% humidity, which indicates that the crystal form A is slightly hygroscopic, and the XRPD result shows that the sample after DVS test has no crystal form change.

Solid state stability influencing factor experiment:

XRPD results show that the crystal form A is stable under the conditions of high temperature (60 ℃), high humidity, illumination and acceleration for 15 days, and crystal form transformation does not occur, wherein the color of a white sample turns yellow slightly under the illumination condition.

TABLE 5 influence factor test results

Crystal form	Condition	7 days result	15 days result
				A	High temperature of 60 DEG C	Without change	Without change
	High humidity 25 deg.C/92.5% RH	Without change	Without change
					Illumination at 25 deg.C/4500 Lux	No change in XFPD, yellowing of the sample	No change in XFPD, yellowing of the sample
	Accelerated by 40 ℃/75% RH	Without change	Without change

TABLE 6 influence factor test results

Through analyzing the data measured by the compound (I), the compound has no obvious change in the A crystal form character and content in the influencing factor experiment and the accelerated experiment, and the maximum single impurity is hardly increased.

Example 5: crystal form a biomedia and water solubility test

The formulation of the biomedia is as follows. Weighing a certain amount of sample, adding the sample into 4.0mL of biological medium or water, shaking for 24h at constant temperature of 37 ℃, sampling for 0.5h, 2h and 24h respectively, filtering the sampled solution by using a 0.22 mu m water-based filter membrane, properly diluting a part of sample with higher concentration by using a diluent, measuring the signal peak area of the solution by using HPLC, and finally calculating the concentration of the compound in the solution according to the peak area, the HPLC standard curve of the raw material and the dilution multiple. In addition, the supernatant was taken for 24h to test its pH and the remaining solids were subjected to XRPD testing.

TABLE 7 arrangement of biological media

The result shows that the 24h solubility of the A crystal form is ranked as FaSSGF > FeSSIF > FaSSIF ≈ water in biological media, and the solubility in the FaSSGF is far higher than that of other media. XRPD results of the remaining solid after solubility testing showed that form a was stable in both biological media and water with no crystal transformation. In addition, the pH of the medium increases after 24 hours of shaking of form a in FaSSGF.

Table 8 dynamic solubility testing in biological media and water

Example 6: thermogravimetric analysis (TGA)

The thermogravimetric analyzer is model number TA Discovery 55(TA, US). 2-5mg of the sample distribution of form a and form B was placed in a balanced open aluminum sample pan and automatically weighed in a TGA furnace. The sample was heated to the final temperature at a rate of 10 deg.C/min with a nitrogen purge rate of 60mL/min at the sample and 40mL/min at the balance. The TGA result shows that the A crystal form has no weight loss in the process of heating to 150 ℃, decomposition is possible to occur above 255 ℃, the TG/DSC spectrogram of the A crystal form compound is shown in figure 2, and the TG/DSC spectrogram of the B crystal form compound is shown in figure 5.

Example 7: differential Scanning Calorimetry (DSC)

The model of the differential scanning calorimetry analyzer is TA Discovery 2500(TA, US). The 1-2mgA crystal form sample is precisely weighed and then placed in a DSC Tzero sample tray with a puncture, the sample is heated to the final temperature at the speed of 10 ℃/min, and the nitrogen purging speed in the furnace is 50 mL/min. DSC result shows that the A crystal form has a melting endothermic peak at about 254 ℃. The NMR results were consistent with the reference spectra. In summary, form a is the anhydrous form (figure 2).

Example 8: x-ray powder diffraction (XRPD)

Samples of the crystalline forms prepared in examples 2 and 3 were analyzed by X-ray powder diffractometry using Bruker D8 Advance (Bruker, GER). The 2 theta scan angle was from 3 deg. to 45 deg., the scan step was 0.02 deg., and the exposure time was 0.08 seconds. When the sample is measured, the voltage and the current of the light tube are respectively 40kV and 40mA, and the sample disc is a zero background sample disc. The XRPD results show that both form a and form B are well crystalline solids. The X-ray diffraction pattern of the crystal form A is shown in figure 1, and the main characteristic peaks of the 2 theta angle are shown in table 9; the X-ray diffraction pattern of the B crystal form is shown in figure 4, and the main characteristic peaks of the 2 theta angle are shown in table 10.

TABLE 9 form A XRPD Main characteristic Peak List

TABLE 10 form B XRPD Main characteristic Peak List

Example 9: polarizing microscope analysis

Polarizing microscope analysis (PLM) the polarizing microscope is of the type Nikon Ci-POL (Nikon, JP). A small amount of A crystal type samples are placed on a glass slide, and proper lenses are selected to observe the appearance of the samples, and the result is shown in figure 8. The resolved structure is consistent with the predicted structure of compound I.

Example 10: single crystal growth and structural analysis of Compound (I), and confirmation of Structure

The purpose of this experiment was to develop single crystal growth and structure analysis of compound (I) and confirm the structure. About 5mg of compound (I) sample was weighed out and dissolved in 1.0mL of methanol/water (v: v 1:1), filtered to obtain a clear solution, the vial containing the clear solution was sealed with a cap, and left to evaporate slowly by one hole at room temperature. After 6 days, rod-like crystals were obtained. The crystal structure was resolved using a Bruker SMART APEX II single crystal X-ray diffractometer.

TABLE 11 Single Crystal Structure analysis data

The compound (I) single crystal belongs to an orthorhombic system, the space group is P212121, and the unit cell parameters are as follows:

α ═ β ═ γ ═ 90.00 ° (see fig. 6 for results). The hydrogen bonds in the crystal cell are intermolecular hydrogen bonds N (1) -H (1A) - - -O (1) and intramolecular hydrogen bonds N (1) -H (1B) - - -O (2), and the crystal molecules are stably arranged in space mainly through the hydrogen bonds and Van der Waals force. Compound (I) single crystal contains 4 molecules of compound (I) in one unit cell (results are shown in fig. 7). Compound (I) single crystal analysis structure and prediction junctionThe structure is consistent.

TABLE 12 anisotropy of temperature factor values of the atoms of the single crystals of Compound (I)

Atom	U11	U22	U33	U23	U13	U12
							O1	0.0554(8)	0.1039(12)	0.0372(7)	0.0162(7)	0.0066(6)	-0.0078(8)
O2	0.0341(5)	0.0465(6)	0.0568(7)	-0.0114(5)	-0.0074(5)	0.0041(5)
							O3	0.0554(7)	0.0561(7)	0.0401(6)	0.0100(5)	-0.0026(5)	0.0179(6)
N1	0.0435(8)	0.0538(8)	0.0468(8)	-0.0016(7)	0.0114(6)	-0.0034(7)
							N2	0.0453(7)	0.0479(7)	0.0348(6)	0.0067(6)	0.0049(6)	0.0165(6)
N3	0.0566(9)	0.0630(9)	0.0395(7)	0.0157(7)	0.0060(6)	0.0081(8)
							N4	0.0778(12)	0.0531(11)	0.0825(12)	-0.0320(10)	-0.0178(10)	0.0098(9)
C1	0.0459(8)	0.0383(8)	0.0355(8)	-0.0030(6)	0.0040(7)	-0.0094(7)
							C2	0.0403(7)	0.0302(7)	0.0280(7)	-0.0026(5)	0.0000(6)	-0.0031(6)
C3	0.0330(7)	0.0321(7)	0.0306(6)	-0.0023(6)	-0.0007(6)	0.0000(6)
							C4	0.0382(8)	0.0328(7)	0.0286(7)	-0.0010(5)	-0.0007(6)	0.0008(6)
C5	0.0441(8)	0.0416(8)	0.0336(7)	0.0023(6)	-0.0024(6)	0.0026(7)
							C6	0.0543(10)	0.0669(11)	0.0390(8)	0.0098(8)	0.0151(8)	0.0077(9)
C7	0.0451(9)	0.0491(9)	0.0382(8)	0.0004(7)	0.0076(7)	0.0059(8)
							C8	0.0417(8)	0.0362(7)	0.0304(7)	0.0005(6)	0.0010(6)	0.0028(6)
C9	0.0468(8)	0.0328(7)	0.0286(7)	-0.0003(6)	-0.0007(6)	0.0001(6)
							C10	0.0627(10)	0.0401(8)	0.0374(8)	0.0060(7)	-0.0013(7)	0.0095(8)
C11	0.0350(7)	0.0312(7)	0.0288(6)	0.0003(5)	0.0013(5)	0.0008(6)
							C12	0.0339(7)	0.0359(7)	0.0327(7)	0.0000(6)	0.0008(6)	0.0007(6)
C13	0.0412(7)	0.0338(7)	0.0397(7)	-0.0014(6)	0.0018(6)	0.0066(6)
							C14	0.0512(9)	0.0320(7)	0.0357(7)	-0.0033(6)	-0.0005(6)	-0.0007(7)
C15	0.0421(8)	0.0411(8)	0.0426(8)	-0.0045(7)	-0.0085(7)	-0.0039(7)
							C16	0.0356(7)	0.0360(7)	0.0389(7)	0.0002(6)	-0.0043(6)	0.0032(6)
C17	0.0568(11)	0.0428(10)	0.0528(10)	-0.0075(8)	-0.0085(8)	0.0019(8)
							C18	0.0417(10)	0.0781(15)	0.1074(19)	-0.0295(14)	-0.0240(12)	0.0184(10)
C19	0.0619(11)	0.0494(9)	0.0460(9)	0.0114(8)	-0.0104(8)	0.0113(9)
							C20	0.0868(16)	0.0718(13)	0.0687(13)	0.0025(12)	-0.0268(13)	0.0008(13)
C21	0.0594(12)	0.0844(15)	0.0541(11)	0.0106(10)	0.0222(10)	0.0254(11)

TABLE 13 Single Crystal Structure analysis data atomic coordinate parameters (x 10)⁴) And equivalent temperature factor value

Atom	x	y	z	U(eq)
					O(1)	6218(2)	1583(2)	10019(1)	66(1)
O(2)	3740(1)	4386(1)	8480(1)	46(1)
					O(3)	6130(2)	4258(1)	7308(1)	51(1)
N(1)	4193(2)	2227(1)	9347(1)	48(1)
					N(2)	9608(2)	1651(1)	8384(1)	43(1)
N(3)	8616(2)	3760(2)	6794(1)	53(1)
					N(4)	6639(3)	7871(2)	10199(1)	71(1)
C(1)	5807(2)	1935(1)	9472(1)	40(1)
					C(2)	7022(2)	2071(1)	8919(1)	33(1)
C(3)	6601(2)	2982(1)	8417(1)	32(1)
					C(4)	7848(2)	2989(1)	7852(1)	33(1)
C(5)	7578(2)	3667(1)	7298(1)	40(1)
					C(6)	10018(3)	3127(2)	6821(1)	53(1)
C(7)	10442(2)	2417(2)	7324(1)	44(1)
					C(8)	9307(2)	2355(1)	7857(1)	36(1)
C(9)	8477(2)	1489(1)	8893(1)	36(1)
					C(10)	9093(3)	629(2)	9377(1)	47(1)
C(11)	6576(2)	4110(1)	8772(1)	32(1)
					C(12)	5129(2)	4782(1)	8799(1)	34(1)
C(13)	5145(2)	5770(1)	9158(1)	38(1)
					C(14)	6608(2)	6085(1)	9489(1)	40(1)
C(15)	8050(2)	5437(1)	9464(1)	42(1)
					C(16)	8011(2)	4456(1)	9107(1)	37(1)
C(17)	6612(3)	7091(2)	9882(1)	51(1)
					C(18)	2295(3)	5068(2)	8448(2)	76(1)
C(19)	5615(3)	4818(2)	6715(1)	52(1)
					C(20)	4880(4)	4051(2)	6223(1)	76(1)
C(21)	12039(3)	1735(2)	7309(1)	66(1)
					H(1A)	3441	2167	9651	58
H(1B)	3916	2472	8964	58
					H(2A)	10549	1294	8395	51
D(3B)	5473	2837	8238	38
					H(6A)	10761	3175	6467	64
H(10A)	8275	538	9722	70
					H(10B)	10145	870	9564	70
H(10C)	9254	-70	9154	70
					H(13A)	4187	6217	9177	46
H(15A)	9025	5657	9683	50
					H(16A)	8975	4014	9090	44
H(18A)	1420	4685	8212	114
					H(18B)	2567	5751	8225	114
H(18C)	1914	5233	8887	114
					H(19A)	6584	5190	6522	63
H(19B)	4789	5386	6825	63
					H(20A)	4554	4466	5839	114
H(20B)	3908	3691	6408	114
					H(20C)	5701	3499	6103	114
H(21A)	12644	1889	6909	99
					H(21B)	11761	956	7327	99
H(21C)	12729	1927	7681	99

Example 11: WKY rat nephropathy model drug effect experiment induced by deoxycorticosterone acetate/sodium chloride load

The objective of this study was to evaluate the efficacy of compounds in a deoxycorticosterone acetate (DOCA)/sodium chloride load-induced WKY rat nephropathy model.

Reagent:

deoxycorticosterone acetate: shanghai Michelin Biochemical technology, Inc., cat #: d830008

Carboxymethyl cellulose: Sigma-Aldrich, Cat number: c4888

Distilled water: drech's series

Tissue fixing liquid: wuxi city Jiangyuan industry technology trade general company, lot number: 201127

High-sodium-salt feed: shanghai Proplung Biotech Co., Ltd., batch number: 20210608-

Microalbumin ELISA detection kit: elabscience, cat #: E-EL-R0362c

Equipment:

a sphygmomanometer: KENT, model number: CODA Monitor-CM2228B

An electronic balance: sartorius, type: quintix124-1CN

Electronic scale: shanghai yuepin scientific instruments ltd, model: YP10001

An ultrasonic cleaner: kunshan ultrasonic instruments ltd, model: KQ3200E

An oscillator: its linbel, model: 01-00X-1012

High-speed refiner: IKA, type: t10 basic

An anesthesia machine: MIP, model: 50318

Freezing a centrifuge: thermo Fisher, cat #: sorvall Legend Micro 17r

A multifunctional microplate reader: TECAN, good number: SPARK 10M

And (3) microscope: OLYMPUS, cat #: DP720

Tissue dehydrator: leica, cat number: TP1020

A paraffin embedding machine: leica, cat number: EG1150H

Freezing: leica, cat number: EG1150C

Slicing machine: leica, cat number: RM2235

Sheet pushing machine: leica, cat number: HI1210

Full-automatic biochemical analyzer: hitachi High-Technologies Corporation, cat # HITACHI-7180(751-

TABLE 14 sample information

Serial number

Sample (I)

Batch number

Traits

Molecular weight

Purity of

Storage conditions

1

Finerenone

20201212

White solid

378.43

>98％

0-4 deg.C, avoiding light

2

Crystal form A of compound (I)

MC-252-114

White solid

379.44

99.59％

0-4 deg.C, avoiding light

TABLE 15 animal information

Remarking: all manipulations of animals in this protocol were approved by the Shanghai Nuno laboratory animal welfare and use administration Committee (IACUC, BD-202103151).

Animal grouping:

the experiment selects SPF-grade male 6-week-old WKY rats which are purchased from Zhejiang Wintonlihua experiment animal technology limited department, and all animals are marked by ear tags. Rats were randomly divided into 6 groups by body weight.

Sample preparation:

(ii) carboxymethyl cellulose (5mg/mL, 100 mL): 0.5g of carboxymethyl cellulose is weighed into a 200mL glass bottle, 100mL of purified water is added, and a clear solution is prepared by stirring.

(ii) deoxycorticosterone acetate (20mg/mL, 50 mL): weighing 1.0g deoxycorticosterone acetate into a 100mL glass bottle, adding 50mL 0.5% carboxymethyl cellulose solution, and preparing uniform suspension by vortex ultrasonic.

Test compounds (formulated for actual animal numbers and body weights, stored at 4 ℃, as exemplified by 40mL per group):

③ menstruum: 1% sodium carboxymethylcellulose solution (10mg/mL, 100 mL): 1.0g of carboxymethyl cellulose is weighed into a 200mL glass bottle, 100mL of purified water is added, and a clear solution is prepared by stirring.

Finorenone (0.2 mg/mL): 8.0mg of Finorenone is weighed into a 60mL glass bottle, 40mL of solvent is added, and vortex ultrasound is carried out to prepare uniform suspension.

Form a of compound (I) (0.4mg/mL) high dose: 28.0mg of the form A of the compound (I) is weighed in a 120mL glass bottle, 70mL of solvent is added, and uniform suspension is prepared by vortex ultrasound.

Sixthly, the dosage of the compound (I) in the crystal form A (0.2 mg/mL): sucking 30mL of the A crystal form suspension of the compound (I) with the concentration of 0.4mg/mL, adding the A crystal form suspension into a 60mL glass bottle, adding 30mL of solvent, and preparing the mixture into a uniform suspension by vortex ultrasonic.

Low dose of form a of compound I (0.1 mg/mL): sucking 20mL of the compound (I) crystal form A suspension of 0.2mg/mL, adding the compound (I) crystal form A suspension into a 60mL glass bottle, adding 20mL of solvent, and performing vortex ultrasonic treatment to prepare a uniform suspension.

Table 16. animal groups:

animal modeling and drug administration treatment:

on the day of the experimental modelling, all animals were divided into 6 groups of 10 animals each according to body weight. In addition to the normal group, WKY rats were subjected to unilateral nephrectomy. After one week, all rats in the model group were given free diet containing 4% sodium chloride (normal feed containing 4% NaCl) and injected with DOCA subcutaneously once a week. 4 weeks after DOCA injection, the doses were administered for 4 weeks as per Table 16. After 4 weeks of dosing (15 weeks of age), all animals were dissected. At 7, 11, 13 and 15 weeks of age, 24 hour urine collection was performed using metabolic cages.

The evaluation indexes are as follows:

weight and kidney weight: body weight was measured before grouping after 1 week of acclimation. Body weight was monitored daily after dosing, body weight and kidney weight were measured at the end of the experiment and kidney weight/body weight was calculated.

Blood pressure: blood pressure was measured at 11 and 15 weeks of age of the animals.

③ urine: when the animal is 15 weeks old, detecting urine albumin by an ELISA method, and detecting urine creatinine by a full-automatic biochemical analyzer; when the animals are 7, 11, 13 and 15 weeks old, the urine protein concentration is measured by a full-automatic biochemical analyzer; when the animals are 11, 13 and 15 weeks old, the concentration of sodium and potassium ions in urine is measured by a full-automatic biochemical analyzer.

-alterations in renal pathology: h & E staining: glomerulosclerosis and renal tubule disorders.

TABLE 17 weight of rats and Kidney weight data

Group of	Number of animals	Endpoint body weight (g)	Kidney mass (mg)	Renal coefficient
					G1
	10	329.6±4.3	1268.2±27.5	0.003847±0.000057
					G2	10	318.2±4.1	2333.6±90.3	0.007321±0.000227
G3	10	316.3±4.0	2132.6±25.0	0.006747±0.000079
					G4	10	312.4±7.0	1990±97.8	0.006398±0.000359
G5	10	314.8±3.8	2009.5±33.7	0.006382±0.000057
					G6	10	307.7±5.9	1931.6±52	0.006274±0.000095

TABLE 18 measurement of urinary albumin and urinary creatinine content in rat

TABLE 19 rat blood pressure measurement results

TABLE 20 Na in rat urine⁺、K⁺Content (wt.)Measurement of

TABLE 21 rat Kidney H & E Pathology score

G1

G2

G3

G4

G5

G6

Glomerular sclerosis index

1.88±0.12

2.75±0.05

2.43±0.09

2.34±0.21

2.33±0.21

2.27±0.20

Number of renal tubular injuries

1.06±0.04

1.65±0.07

1.65±0.09

1.53±0.27

1.09±0.07

1.19±0.14

After 4 weeks of treatment, both the high, medium and low doses of the crystalline form a of Finerenone and compound (I) had a tendency to improve blood pressure, and the crystalline form a of Finerenone and medium dose compound (I) was able to significantly improve the increase in blood pressure in DOCA and 4% high salt feed-induced kidney disease in WKY rats. After 4 weeks of treatment, the three high, medium and low doses of form a of compound (I) significantly enhanced the sodium-excreting and potassium-retaining ability of rats, maintained the Na/K balance in rats to some extent (see the results in fig. 9), and significantly reduced the urinary protein level and albuminuria creatinine ratio. Compared with the normal group of rats, the number and the degree of glomerular sclerosis of the rats in the model group are obviously increased. Compared with the model group, the treatment groups of G3 and G6 obviously reduce the hardening degree of rat glomeruli caused by DOCA and high-sodium feed, and both the groups of G4 and G5 have an improvement trend. Compared with the normal group of rats, the degree of renal tubular injury of the model group of rats is obviously increased, compared with the model group, the treatment groups of G5 and G6 obviously improve the renal tubular injury caused by DOCA and high-sodium feed, and the treatment effect is better than that of the control group and the reference compound Finorenone treatment group G3 (the picture of the renal tissue section is shown in the attached figure 10). The pharmacodynamic experiment result of the WKY rat nephropathy model induced by the deoxycorticosterone acetate (DOCA)/sodium chloride load shows that the A crystal form of the compound (I) has a non-inferior effect on treating the WKY rat nephropathy compared with the Finorenone, and has potential clinical value.

All patents, patent applications, and literature references cited in this specification are hereby incorporated by reference in their entirety. In case of inconsistencies, the present disclosure, including definitions, will be convincing.

Claims (18)

1. A crystal form A of a compound I (4S) -4- (4-cyano-2-methoxyphenyl) -5-ethoxy-2, 8-dimethyl-1, 4-dihydro-1, 6-naphthyridine-4-deuterium-3-formamide shown as a formula I,

the method is characterized in that: the A crystal form X-ray diffraction pattern of the compound I has main characteristic peaks with peak maximum values at 2 theta angles of 8.547 degrees, 11.400 degrees, 14.060 degrees, 15.392 degrees, 19.017 degrees, 19.763 degrees, 22.939 degrees, 23.904 degrees, 25.562 degrees and 26.467 degrees.

2. Form a according to claim 1, characterized in that: the A crystal form infrared spectrogram (v) of the compound_max) At 3475.6, 3417.7, 3364.7, 2990.8, 2952.2, 2939.6, 2229.1, 1682.4, 1657.2, 1607.1, 1574.5, 1483.1, 1432.2, 1324.7, 1169.8, 1031.3, 875.3cm^-1The main characteristic peak with the maximum value of the peak.

3. A process for preparing form a of compound I according to claim 1, comprising the steps of:

s1, mixing a compound I with an inert solvent, and stirring at the temperature of 20-120 ℃;

s2, dropwise adding a benign solvent until the solid is just completely dissolved, cooling the solution to 0-30 ℃, and precipitating a crystal form A;

s3, after filtering, drying a filter cake in vacuum at the temperature of 35 +/-5 ℃;

the benign solvent comprises dimethyl sulfoxide, methanol, acetone, ethylene glycol methyl ether, dimethylformamide, dichloromethane, chloroform, ethanol, n-propanol, isopropanol, 4-methyl-2-pentanone, ethyl acetate, ethyl formate, butyl formate, dioxane, ethylene glycol dimethyl ether and acetonitrile.

4. The method of claim 3, wherein: the inert solvent comprises isopropyl acetate, n-heptane, cyclohexane, diethyl ether, methyl tert-butyl ether, water and toluene.

5. The method of claim 3, wherein: compound I in step S1 is one or more polymorphs or solvates thereof.

6. A crystalline form B of compound I characterized by: the X-ray diffraction pattern of the B crystal form of the compound I has main characteristic peaks with peak maximum values at 2 theta angles of 5.792 degrees, 10.273 degrees, 10.635 degrees, 11.267 degrees, 12.048 degrees, 14.834 degrees, 21.849 degrees, 22.535 degrees, 24.171 degrees and 24.734 degrees.

7. A process for preparing form B of compound I according to claim 6, comprising the steps of:

s1, mixing a compound I with an inert solvent, and stirring at the temperature of 20-120 ℃;

s2, adding tetrahydrofuran dropwise until the solid is just completely dissolved, cooling the solution to 0-30 ℃, and precipitating a crystal form B;

s3, after filtering, drying a filter cake in vacuum at the temperature of 35 +/-5 ℃.

8. The process of claim 7, wherein the inert solvent comprises isopropyl acetate, n-heptane, cyclohexane, diethyl ether, methyl tert-butyl ether, water, toluene.

9. The method of claim 7, wherein the solution cooling temperature of step S2 is 0-5 ℃.

10. Use of the crystalline form a of claim 1 or the crystalline form B of claim 5 in the manufacture of a medicament for the treatment of diabetic nephropathy and cardiovascular disorders.

11. A process for the preparation of compound X, characterized in that it comprises the following steps:

1. the compound XIII is subjected to catalytic cyanation reaction of metal palladium to obtain a compound XIV;

2. compound X was prepared by deuterating compound XIV under the action of compound XVI.

12. The synthesis method of claim 11, wherein in step 1, compound XIII, potassium hexachloroferricyanide and sodium carbonate are reacted in N, N-dimethylformamide under the catalysis of palladium acetate at 120 ℃ under nitrogen atmosphere for 12-15 hours to obtain compound XIV.

13. The method of claim 11, wherein: in the step 2, firstly, reacting a compound XV and triethyl orthoformate in a mixed solution of acetic acid and water at room temperature for 12-15 hours, adding water and ammonium tetrafluoroborate, and stirring at room temperature for 30-50 minutes to obtain a compound XVI; then compound XVI and compound XIV react with sodium bicarbonate in toluene and deuterated heavy water at 100 ℃ under nitrogen atmosphere for 5-8 hours to obtain compound X.

14. A process for the preparation of compound I, characterized by comprising the steps of:

1. the compound (R, S) -I is reacted with chiral tartaric acid dibenzoate to obtain a compound XVII;

2. compound XVII is liberated by addition of base to give compound I.

15. The method of claim 14, wherein: the compound (R, S) - (I) is prepared by the one-pot reaction of firstly generating carbonyl imidazole from the compound IV under the action of a condensation agent carbonyl diimidazole, and then hydrolyzing through amidation reaction with hexamethyldisilazane.

16. The method of claim 15, wherein: the preparation method of the compound (R, S) - (I) comprises the following steps: compound IV is first reacted with diimidazole carbonate and a catalytic amount of 4- (dimethylamino) pyridine in THF to produce a carbonylimidazole intermediate; then adding hexamethyldisilane into the reaction liquid to generate N-silane amide; finally hydrolyzing to obtain the compound (R, S) -I.

17. The method of claim 15, wherein: the preparation method of the compound IV comprises the following steps:

1. compound XI is prepared by a boronic acid catalyzed transesterification reaction to compound IX;

2. compound IX and compound X prepared by the preparation method according to claim 11 are subjected to Knoevenagel dehydration condensation reaction to obtain a mixture of compound VIIa and compound VIIb;

3. preparing a compound VI by a condensation cyclization reaction of a mixture of a compound VIIa and a compound VIIb and a compound VIII;

4. carrying out etherification reaction on the compound VI to obtain a compound V;

5. compound V is hydrolyzed to produce compound IV.

18. The preparation method of the compound I is characterized in that the synthetic route is as follows:

the preparation method comprises the following steps:

step 1, reacting a compound XI and a compound XII with boric acid in toluene at 110 ℃ to obtain a compound IX;

step 2, adding palladium acetate into a compound XIII, potassium hexachloroferricyanide and sodium carbonate in N, N-dimethylformamide for catalysis, and reacting for 12-15 hours at 120 ℃ under nitrogen atmosphere to obtain a compound XIV;

step 3, firstly, reacting a compound XV and triethyl orthoformate in a mixed solution of acetic acid and water at room temperature for 12-15 hours, adding water and ammonium tetrafluoroborate, and stirring at room temperature for 30-50 minutes to obtain a compound XVI; then the compound XVI and the compound XIV react with sodium bicarbonate in toluene and deuterated heavy water at the external temperature of 100 ℃ under nitrogen atmosphere for 5 to 8 hours to obtain a compound X;

step 4, reacting compound IX and compound X with acetic acid and piperidine in isopropanol at room temperature to obtain a mixture of compound VIIa and compound VIIb;

step 5, reacting a mixture of a compound VIIa and a compound VIIb with a compound VIII in an ethanol solution at 95 ℃ to obtain a compound VI;

step 6, adding concentrated sulfuric acid into the compound VI and triethyl orthoacetate in NMP for catalysis, and reacting at 120 ℃ to obtain a compound V;

step 7, the volume ratio of compound V to aqueous sodium hydroxide solution in THF/water is 2:1 at 0 ℃ to obtain a compound IV;

step 8, compound IV is first reacted with diimidazole carbonate and a catalytic amount of 4- (dimethylamino) pyridine in THF to produce a carbonylimidazole intermediate; then adding hexamethyldisilane into the reaction liquid to generate N-silane amide; finally hydrolyzing to obtain a compound (R, S) -I;

step 9 and step 10, the compound (R, S) -I is resolved into compound I and/or compound Ia by using compound IIIa or IIIb.

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