WO2017121323A1 - Preparation method for pyridine derivative compound, intermediate and crystal form thereof - Google Patents

Abstract

A preparation method for a salt of a compound (1R,2S)-1-(5-(4-chlorophenyl)-2-methoxypyridin-3-yl)-4-dimethylamino-2-aryl-1-phenylbutan-2-ol of formula (I) useful for resisting Mycobacterium tuberculosis, a crystal form, an intermediate compound and a preparation method therefor.

Description

Preparation method of pyridine derivative compound, intermediate and crystal form thereof Technical field

The present invention relates to a high purity (1R,2S)-1-(5-(4-chlorophenyl)-2-methoxypyridin-3-yl)-4-dimethylamino-2-aryl-1 A process for the preparation of a salt of phenylbutan-2-ol and a crystalline form thereof, and to an intermediate compound for the preparation of a compound of the formula (I) and a process for the preparation thereof.

Background technique

Mycobacterium tuberculosis is the causative agent of tuberculosis. As a globally widespread and fatal infectious disease, according to the World Health Organization, more than 8 million people are infected each year and 2 million die from tuberculosis. In the past decade, tuberculosis cases have grown at a rate of 20% worldwide, especially in poor areas. If this trend continues, tuberculosis cases are likely to continue to grow at a 41% increase over the next two decades. In the 50 years since the initial application of chemotherapy, tuberculosis has been the leading infection to adults, second only to AIDS. Complications of tuberculosis have led to the emergence of many drug-resistant strains and a symbiotic relationship with AIDS. People who tested positive for HIV and who were infected with tuberculosis had a 30-fold greater chance of developing activated tuberculosis than those who were negative for the HIV test. On average, one out of every three patients who die of AIDS is caused by tuberculosis.

The current treatment of tuberculosis employs a combination of multiple agents. For example, a formula recommended by the US Department of Public Health consists of first using isoniazid, rifampicin, pyrazinamide, and ethambutol for two months, then using isoniazid and rifampin alone for four months. . For patients infected with AIDS, the use of this combination of drugs needs to be extended to seven months. For patients infected with multidrug-resistant tuberculosis, the drug combination also needs to add the remaining agents, such as ethambutol tablets, streptomycin, kanamycin, amikacin, capreomycin, ethionamide, Cycloserine, ciprofloxacin and ofloxacin.

For the benefit of patients and suppliers, new therapies that can improve current treatments are highly desirable, such as shorter treatment cycles and less need for supervised treatments. In the first two months of treatment, the mixed four drugs concentrated on the bacteria, thereby greatly reducing the number of bacteria and making the patient less infectious. In the next 4 to 6 months, the bacteria in the patient are eliminated, reducing the likelihood of recurrence. A powerful bactericidal drug that reduces the treatment cycle to two months or less can bring huge benefits. At the same time, the drug should require less supervision. Obviously, drugs that both shorten the treatment time and reduce the frequency of supervision can bring the greatest benefit.

Complications of infectious tuberculosis cause multi-drug resistant tuberculosis. Worldwide, 4% of cases are related to multi-drug resistant tuberculosis. Multi-drug resistant tuberculosis is primarily resistant to isoniazid and rifampicin in four standard treatments. Multi-drug resistant tuberculosis can be fatal if there is no treatment or if standard tuberculosis standard therapy is used. Therefore, the treatment of this disease requires the use of second-line drugs for two years. Most of these second-line drugs are toxic, expensive, and have low efficacy. Infectious drug-resistant tuberculosis patients continue to spread the disease due to the lack of effective treatment. Therefore, for multi-drug resistant tuberculosis, a new drug with a novel mechanism of action is highly demanded.

The application number: 201410335196.X describes a new class of pyridine derivatives and their use as antimycobacteria for the treatment of tuberculosis, especially multi-drug resistant tuberculosis. Its structure is as shown in formula (B-1):

Summary of the invention

The present invention provides a process for the preparation of a compound of formula (I),

It contains the following steps:

among them,

R ^{1 is} selected from a 6- to 12-membered aryl group, a 6- to 12-membered heteroaryl group, a 6- to 12-membered aryl-alkylene group, and a 6 to 12-membered heterocyclic group, which are optionally substituted by 0, 1, 2 or 3 R ⁰¹ . Aryl-alkylene;

HX is selected from an organic or inorganic acid;

The base A is selected from the group consisting of an alkali metal base, an alkaline earth metal base or an organometallic base;

The molar ratio of the compound (II) to the base A is 1:1 to 5;

The molar ratio of the compound (II) to the compound (III) is 1:1 to 2;

The reaction solvent is selected from a single ether solvent or a mixed solvent of several ether solvents;

The amount of the reaction solvent is 3 to 20 times the weight of the compound (IV);

The reaction temperature is -80 to 0 ° C;

The reaction time is 1 to 24 hours;

R ^{01 is} selected from the group consisting of F, Cl, Br, I, CN, OH, CH(CH ₃ ) ₂ , C(CH ₃ ) ₃ , N(CH ₃ ) ₂ , NH(CH ₃ ), NH ₂ , CHO, COOH, C(=O)NH ₂ , S(=O)NH ₂ , S(=O) ₂ NH ₂ , CF ₃ , CF ₃ O, (NH ₂ )CH ₂ , (HO)CH ₂ , CH ₃ C(= O), CH ₃ OC(=O), CH ₃ S(=O) ₂ , CH ₃ S(=O); the "hetero" represents a hetero atom selected from N, O or S;

The number of heteroatoms is independently selected from 1, 2 or 3, respectively.

In some embodiments of the invention, the above R ¹ is a naphthyl or phenyl group optionally substituted with 0, 1, 2 or 3 R ⁰¹ .

In some aspects of the invention, the above R ^{1 is} selected from the group consisting of

In some aspects of the invention, the alkali metal base is selected from the group consisting of lithium hydroxide, sodium hydroxide, potassium hydroxide, barium hydroxide, sodium carbonate, potassium carbonate, barium carbonate, sodium hydrogencarbonate, and/or potassium hydrogencarbonate.

In some embodiments of the invention, the alkaline earth metal base is selected from the group consisting of sodium hydride, potassium hydride, and/or calcium hydride.

In some aspects of the invention, the organometallic base is selected from the group consisting of n-butyl lithium, lithium diisopropylamide, lithium 2,2,6,6-tetramethylpiperidine, lithium bis(trimethylsilyl)amide, Sodium methoxide, lithium t-butoxide, sodium t-butoxide, potassium t-butoxide, sodium ethoxide and/or aluminum isopropoxide.

In some embodiments of the invention, the molar ratio of the compound (II) to the base A is 1:1.2 to 2.

In some embodiments of the invention, the above reaction temperature is from -80 to -60 °C.

In some aspects of the invention, the reaction time is from 2 to 12 hours.

In some aspects of the invention, the reaction time is from 4 to 8 hours.

In some embodiments of the invention, the above reaction solvent is selected from the group consisting of tetrahydrofuran, diethyl ether and/or isopropyl ether.

In some aspects of the invention, the amount of the above reaction solvent is 5 to 10 times the weight of the compound (IV).

In some aspects of the invention, the above preparation method further comprises the following reaction route:

In some aspects of the invention, the above preparation method further comprises the following reaction route:

In some aspects of the invention, the above preparation method further comprises the following reaction route:

among them,

The base B is selected from the group consisting of lithium hydroxide, sodium hydroxide, potassium hydroxide, barium hydroxide, sodium carbonate, potassium carbonate, barium carbonate, sodium hydrogencarbonate, potassium hydrogencarbonate, sodium methoxide, lithium t-butoxide, sodium t-butoxide, Potassium tert-butoxide, sodium ethoxide, aluminum isopropoxide;

The reaction solvent is selected from a mixed solvent of a ketone, an alcohol or an ester solvent and a polar aprotic solvent.

The molar ratio of the chiral acid to the compound (IV) is from 0.5 to 1.5;

The chiral acid is selected from the group consisting of α-hydroxypropionic acid, α-hydroxysuccinic acid, α,β-dihydroxysuccinic acid, α-hydroxyphenylacetic acid, β-hydroxy acid, and compound (VI);

n is 0, 1 or 2;

R ² and R ⁴ are each independently selected from H, F, Cl, Br, I, or selected from, optionally substituted by 0, 1, 2 or 3 R ⁰¹ : C _1-8 alkoxy, C _{1- 8-} alkyl, Si(Ph) ₃ , 6- to 12-membered aryl;

R ³ and R ⁵ are each independently selected from H, F, Cl, Br, I, NO ₂ , OH, or selected from C _1-8 alkoxy optionally substituted by 0, 1, 2 or 3 R ⁰¹ a group, a C _1-8 alkyl group, a 6 to 12 membered aryl group;

Optionally, position 13 and position 14 is substituted with R ³ or at position 14 and position 15 is substituted with R ³ may be joined together to form a 6 to 12 membered aryl ring;

Optionally, the 8 position and R ⁵ is substituted at position 9 or position 9 and joined together at position 10 substituted with R ^5, form a 6-12 membered aryl ring.

In some embodiments of the invention, the molar ratio of the chiral acid to the compound (IV) is from 0.8 to 1.2.

In some embodiments of the invention, the molar ratio of the chiral acid to the compound (IV) is 1.0.

In some embodiments of the present invention, in the above process for producing the compound (V), in the reaction solvent, the ketone solvent is selected from the group consisting of acetone and/or methyl ethyl ketone.

In some embodiments of the invention, in the above method for producing the compound (V), in the reaction solvent, the alcohol solvent is selected from the group consisting of ethanol, methanol, isopropanol and/or tert-butanol.

In some embodiments of the present invention, the above process for producing the compound (V), wherein the ester solvent is selected from the group consisting of ethyl acetate and/or t-butyl acetate.

In some embodiments of the invention, the above process for preparing compound (V), wherein the polar aprotic solvent is selected from the group consisting of DMF, DMSO, DMA and/or NMP.

In some aspects of the invention, the method for producing the compound (V), wherein the mixed solvent is a mixed solvent of ethanol and DMF or a mixed solvent of ethanol and DMSO.

In some embodiments of the invention, the solvent for preparing compound (V) from compound (IV) is selected from the group consisting of acetone, methyl ethyl ketone, ethanol, methanol, isopropanol, tert-butanol, ethyl acetate, t-butyl acetate A single solvent or a mixed solvent of several solvents in DMF, DMSO, DMA and/or NMP.

In some aspects of the invention, the above method for producing the compound (V), wherein the volume ratio of the ketone, alcohol or ester solvent to the polar aprotic solvent is 1:0.03 to 0.1.

In some aspects of the invention, the above process for the compound (V), wherein the solvent is used in an amount of from 15 to 50 times the weight of the compound (IV).

In some embodiments of the invention, the above R ³ and R ⁵ are each independently selected from H.

In some embodiments of the invention, the above R ^{2 is} substituted at the 2-position.

In some embodiments of the invention, the above R ^{4 is} substituted at the 6 position.

In some aspects of the present invention, the above R ² and R ⁴ are each independently selected from the group consisting of: H, Si(Ph) ₃ ,

In some embodiments of the invention, the above compound (VI) is selected from the group consisting of

In some aspects of the invention, the above preparation method further comprises the following reaction route:

Wherein HX is selected from an organic or inorganic acid.

In some embodiments of the invention, the HX is selected from the group consisting of hydrochloric acid, sulfuric acid, phosphoric acid, oxalic acid, citric acid, maleic acid or fumaric acid.

In some aspects of the invention, the above preparation method further comprises the following reaction route:

The present invention provides a compound of the formula: as an intermediate for the preparation of a compound of formula (I):

The present invention also provides a compound I-1 represented by the following formula,

The present invention provides a crystalline form I of Compound I-1, the XRPD pattern of which is shown in Figure 1.

In some aspects of the present invention, the XRPD pattern analysis data of the I crystal form is shown in Table-1.

Table-1 I crystal form XRPD pattern analysis data

The present invention provides a crystalline form II of Compound I-1, the XRPD pattern of which is shown in Figure 4.

In some aspects of the present invention, the XRPD pattern analysis data of the above II crystal form is shown in Table-2.

Table-2 XRPD pattern analysis data of II crystal form

NO.	2-Theta	d(A)	I%	NO.	2-Theta	d(A)	I%
1	4.903	18.0073	22.0	5	12.263	7.2114	12.2
2	8.595	10.2792	100.0	6	12.778	6.9219	24.8
3	10.959	8.0665	12.5	7	15.560	5.6902	15.7
4	11.906	7.4269	14.1	8	21.700	4.0920	7.0

The present invention provides a crystalline form III of Compound I-1, the XRPD pattern of which is shown in FIG.

In some aspects of the present invention, the XRPD pattern analysis data of the above III crystal form is shown in Table-3.

Table-3 XRPD pattern analysis data of III crystal form

The invention provides a preparation method of the above crystal form I, which comprises preparing a compound I-1 of any one form by adding to a solvent, wherein the solvent is selected from the group consisting of an alcohol, a ketone solvent or an alcohol solvent and a ketone solvent. a mixed solvent; the solvent is used in an amount of from 3 to 50 times the weight of the compound I-1.

In some aspects of the invention, in the method for preparing the above crystal form I, the alcohol solvent is selected from the group consisting of methanol, ethanol, isopropanol and/or n-butanol.

In some aspects of the invention, in the method for preparing the above crystal form I, the ketone solvent is selected from the group consisting of acetone and/or methyl ethyl ketone.

In some aspects of the invention, in the method for preparing the above-mentioned Form I, the mixed solvent is a mixed solvent of methanol and acetone.

In some aspects of the invention, in the method for preparing the above-mentioned Form I, the volume ratio of the mixed solvent of methanol and acetone is 1:5 to 30.

Definition and description:

Unless otherwise stated, the following terms and phrases used herein are intended to have the following meanings. A particular phrase or term should not be considered undefined or unclear without a particular definition, but should be understood in the ordinary sense. When a trade name appears in this document, it is intended to refer to its corresponding commodity or its active ingredient.

The intermediate compounds of the present invention can be prepared by a variety of synthetic methods well known to those skilled in the art, including the specific embodiments listed below, combinations thereof with other chemical synthesis methods, and those skilled in the art. Well-known equivalents, preferred embodiments include, but are not limited to, embodiments of the invention.

The chemical reaction of a particular embodiment of the invention is carried out in a suitable solvent which is suitable for the chemical changes of the invention and the reagents and materials required thereof. In order to obtain the compounds of the present invention, it is sometimes necessary for those skilled in the art to modify or select the synthetic steps or reaction schemes based on the prior embodiments.

The invention is specifically described by the following examples, which are not intended to limit the invention.

All solvents used in the present invention are commercially available and can be used without further purification. The reaction is generally carried out under an inert nitrogen atmosphere in an anhydrous solvent. Proton nuclear magnetic resonance data was recorded on a Bruker Avance III 400 (400 MHz) spectrometer with chemical shifts expressed in ppm at the low field of tetramethylsilane. Mass spectra were measured on an Agilent 1200 Series Plus 6110 (&1956A). LC/MS or Shimadzu MS contains a DAD: SPD-M20A (LC) and Shimadzu Micromass 2020 detector. The mass spectrometer is equipped with an electrospray ionization source (ESI) operating in either positive or negative mode.

The present invention employs the following abbreviations: DMF stands for N,N-dimethylformamide; DMA stands for N,N-dimethylacetamide; DMSO stands for dimethyl sulfoxide; NMP stands for N-methylpyrrolidone; Pd( OAc) ₂ represents palladium acetate; Pd(dppf)Cl ₂ represents [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride; Pd ₂ (dba) ₃ represents tris(dibenzylideneacetone) Palladium; Pd(PPh ₃ ) ₄ represents tetrakistriphenylphosphine; Pd(PPh ₃ ) ₂ Cl ₂ represents dichloroditriphenylphosphine palladium; Et ₃ N represents triethylamine; DIPEA stands for diisopropyl Ethylamine; DBU stands for 1,8-diazabicycloundec-7-ene; AcOH stands for glacial acetic acid; Na ₂ SO ₃ stands for sodium sulfite; MeOH stands for methanol; TMP stands for 2,2,6,6-four Methylpiperidine; pH represents the hydrogen ion concentration index.

Compound by hand or

Software naming, commercially available compounds using the supplier catalog name.

X-ray powder diffractometer (XRPD)

Instrument model: Brooke D8advance X-ray diffractometer

Test conditions: Detailed XRPD parameters are as follows:

X-ray generator: Cu, kα,

Tube voltage: 40kV, tube current: 40mA.

Launch slit: 1deg.

Height limit slit: 10mm

Scattering slit: 1deg.

Accept the slit: 0.15mm

Monochromator: fixed monochromator

Scan range: 4-40deg.

Scanning speed: 10deg/min

Differential Scanning Calorimeter (DSC)

Instrument model: TA Q2000 Differential Scanning Calorimeter

Test conditions: The sample (~1 mg) was placed in a DSC aluminum pan for testing at 25 ° C - 300 ° C and a heating rate of 10 ° C / min.

Thermo Gravimetric Analyzer (TGA)

Instrument model: TA Q5000IR thermogravimetric analyzer

Test conditions: Samples (2 to 5 mg) were placed in a TGA platinum pot for testing at room temperature - 300 ° C and a heating rate of 10 ° C / min.

Beneficial effects:

The compound I-1 provided by the present invention has stable properties, good solubility, good wettability, and good crystallinity, crystal form and crystal form III. Prospects for medicine.

The process for synthesizing the compound I-1 and the intermediate thereof provided by the invention overcomes the disadvantages of the prior art that the starting materials are expensive, the reagents used are poisonous, the reaction conditions are harsh, the separation and purification are difficult, and the industrialization is difficult.

specifically:

1) The method for preparing the compound I-1 of the present invention is a conventional or common reagent, which is easily available on the market and is inexpensive;

2) The intermediate compound (II) can be obtained from the compound (b) by a two-step conventional reaction in a higher total yield, and the post-treatment is simple without any column chromatography purification;

3) kinetic resolution of compound (IV) by using an organic acid, and the obtained compound (I) has high optical purity;

4) The reagents used in the reaction of each step are small molecules and easy to be purified;

5) The metal palladium catalyzed cross-coupling is placed in a relatively advanced step, which is beneficial to control the metal palladium residue in the final product.

DRAWINGS

Figure 1 is an XRPD spectrum of Form I Cu-Kα radiation of Compound I-1.

Figure 2 is a DSC chart of Form I of Compound I-1.

Figure 3 is a TGA pattern of Form I of Compound I-1.

4 is an XRPD spectrum of Form II Cu-Kα radiation of Compound I-1.

Figure 5 is a DSC chart of Form II of Compound I-1.

Figure 6 is a TGA pattern of Form II of Compound I-1.

Figure 7 is an XRPD spectrum of Form III Cu-Kα radiation of Compound 1-1.

Figure 8 is a DSC chart of Form III of Compound I-1.

Figure 9 is a TGA pattern of Form III of Compound I-1.

Detailed ways

For a better understanding of the content of the present invention, the following detailed description is made in conjunction with the specific embodiments, but the specific embodiments are not limited to the content of the present invention.

Compound 1-(5-(4-chlorophenyl)-2-methoxypyridin-3-yl)-4-(dimethylamino)-2-(naphthalen-1-yl)-1-phenyl The-2-ol has the following structure:

This compound has two chiral centers, as indicated by * in formula (IV-1), and thus has four stereoisomers, respectively (1R, 2S) 1-(5-(4-chlorophenyl)-2 -Methoxypyridin-3-yl)-4-(dimethylamino)-2-(naphthalen-1-yl)-1-phenylbutan-2-ol, (1S,2R)1-(5- (4-chlorophenyl)-2-methoxy Pyridin-3-yl)-4-(dimethylamino)-2-(naphthalen-1-yl)-1-phenylbutan-2-ol, (1R, 2R) 1-(5-(4-chloro Phenyl)-2-methoxypyridin-3-yl)-4-(dimethylamino)-2-(naphthalen-1-yl)-1-phenylbutan-2-ol and (1S,2S)1 -(5-(4-chlorophenyl)-2-methoxypyridin-3-yl)-4-(dimethylamino)-2-(naphthalen-1-yl)-1-phenylbutene-2 -alcohol. These four stereoisomers are composed of two sets of diastereomers, wherein the racemic mixture (1R, 2S) 1-(5-(4-chlorophenyl)-2-methoxypyridine- 3-yl)-4-(dimethylamino)-2-(naphthalen-1-yl)-1-phenylbutan-2-ol and (1S,2R)1-(5-(4-chlorophenyl) 2-methoxypyridin-3-yl)-4-(dimethylamino)-2-(naphthalen-1-yl)-1-phenylbutan-2-ol will be labeled as "A"; and the racemic mixture (1R, 2R) 1-(5-(4-chlorophenyl)-2-methoxypyridin-3-yl)-4-(dimethylamino)-2 -(naphthalen-1-yl)-1-phenylbutan-2-ol and (1S,2S)1-(5-(4-chlorophenyl)-2-methoxypyridin-3-yl)-4- (Dimethylamino)-2-(naphthalen-1-yl)-1-phenylbutan-2-ol will be designated "B" in the Examples section that follows.

The examples set forth below are all prepared, separated and characterized by the methods described herein. The following examples are merely representative of the scope of the invention, and not all of them. The present invention has been described in detail herein, the embodiments of the present invention are disclosed herein, and various modifications and changes may be made to the embodiments of the present invention without departing from the spirit and scope of the invention. It will be obvious.

Example 1: Preparation of Compound I-1

Process 1:

Step 1: Synthesis of 5-(4-chlorophenyl)-2-methoxypyridine

Potassium carbonate (5.51 kg, 39.89 moles, 1.5 equivalents) and Pd(dppf)Cl ₂ (48.64 g, 66.48 mmol, 0.0025 equivalents) were added to 5-bromo-2-methoxypyridine (5.0, respectively) with stirring. Kilogram, 26.59 moles, 1.0 equivalents) and 4-chlorophenylboronic acid (4.49 kg, 28.72 moles, 1.08 equivalents) in a mixed solution of dioxane (25 liters) and water (5 liters), replaced with nitrogen three times, then The reaction system was heated to 95 to 100 ° C, and the reaction was refluxed at this temperature for 16 hours. The reaction was stopped after the end of consumption of 5-bromo-2-methoxypyridine by HPLC. The reaction solution was cooled to 25 to 30 ° C, and concentrated under reduced pressure to remove dioxane. The residue was extracted twice with ethyl acetate (10)升 × 2), the organic phase was combined, washed with saturated brine (5 liters × 2), concentrated under reduced pressure at 35-40 ° C to give crude 5-(4-chlorophenyl)-2-methoxypyridine ( 6.5 kg, dark brown needle-like solid after cooling), the crude product was directly used for the next reaction. _{^{1 H NMR (400MHz, CDCl 3}} ) δ: 8.37 (d, J = 2.1Hz, 1H), 7.75 (dd, J = 8.6,2.6Hz, 1H), 7.48-7.39 (m, 4H), 6.83 (d, J = 8.6 Hz, 1H), 4.00 (s, 3H).

Step 2: Synthesis of 3-bromo-5-(4-chlorophenyl)-2-methoxypyridine

5-(4-Chlorophenyl)-2-methoxypyridine (6.5 kg, 29.59 mol, 1.00 eq.) was dissolved in DMF (21 L) at 25 ° C for 4 to 5 hours, slowly added dropwise. To a solution of liquid bromine (11.82 kg, 73.97 mol, 2.50 eq) in AcOH (7 L). After the completion of the dropwise addition, the reaction mixture was stirred at 25 ° C for 72 hours, TLC (petroleum ether) was used to detect about 20% of the remaining material, and liquid bromine (2.5 kg, 0.5 equivalent) was further added to the reaction system, and stirring was continued for 48 hours. A small amount of raw material (about 7-9%) was detected to be completely unreacted. The reaction was stopped and the reaction solution was slowly added to a saturated Na ₂ SO ₃ (6 kg) aqueous solution with stirring, and the quenching process was controlled to a temperature below 30 °C. A large amount of pale yellow solid was precipitated, filtered, and the filter cake was washed twice with water (20 liters × 2). The obtained crude product was pulverized twice with MeOH (15 liters × 2), filtered, and the filter cake was dried in vacuo (50 ° C). 3-bromo-5-(4-chlorophenyl)-2-methoxypyridine (6.7 kg, white solid, mp. ¹ H NMR (400 MHz, CDCl ₃ ) δ: 8.28 (d, J = 2.3 Hz, 1H), 8.00 (d, J = 2.3 Hz, 1H), 7.43 (s, 4H), 4.06 (s, 3H).

Step 3: Synthesis of 3-benzyl-5-(4-chlorophenyl)-2-methoxypyridine

Preparation of benzyl zinc reagent: Zinc powder (2.85 kg, 43.56 mol, 2.00 eq.) was suspended in anhydrous tetrahydrofuran (21.78 liters) under nitrogen atmosphere, and 1,2-dibromoethane was added at room temperature. (33 ml, 0.02 eq.), then heated to reflux, slowly added trimethylchlorosilane (28 mL, 0.01 eq.) (Note: A large amount of bubbles were observed after the addition and the reflux was severe, if not observed The bubbles were generated, and trimethylchlorosilane was added, and refluxed at 66 ° C for 30 minutes. The reaction solution was then cooled to 25 ° C, and benzyl bromide (3.73 kg, 2.59 liters, 21.78 moles, 1.00 equivalents) was slowly added dropwise to the reaction system (note: the dropping rate was such that the temperature of the system did not exceed 30 ° C) at this temperature. Stirring was continued for 3 hours to obtain a tetrahydrofuran solution of benzyl zinc reagent (concentration: 1 mol/liter), which was used directly for the next reaction.

A solution of 3-bromo-5-(4-chlorophenyl)-2-methoxypyridine (5 kg, 16.75 mol, 1.00 equiv) in tetrahydrofuran (16 L) at room temperature over 30 to 45 under nitrogen. Add to the freshly prepared benzyl zinc reagent tetrahydrofuran solution (1 mol/L, 21.78 L, 1.3 eq.) in a minute, then add Pd(PPh ₃ ) ₂ Cl ₂ (59 g, 83.75 mmol, 0.005 equivalent). The solution was stirred at room temperature for 16 hours under a nitrogen atmosphere (the reaction system itself exothermed, the temperature was raised to 50 ° C, and then slowly cooled, and the reaction liquid turned grayish black). TLC (petroleum ether / ethyl acetate = 20/1) was obtained. The reaction mixture was filtered. EtOAc (EtOAc m. The suspended organic phase was filtered again, and the filtrate was washed twice with saturated brine (10 liters×2), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure at 35 to 40 ° C to give a brown oil. Slowly pour into methanol (10 liters), a large amount of white precipitate was precipitated, filtered, and the cake was beaten with methanol (2 liters × 2) to give 3-benzyl-5-(4-chlorophenyl)-2- Methoxypyridine (3.0 kg, white solid, yield 58%). _{^{1 H NMR (400MHz, CDCl 3}} ) δ: 8.23 (d, J = 2.4Hz, 1H), 7.47 (d, J = 2.4Hz, 1H), 7.42-7.35 (m, 4H), 7.35-7.29 (m, 2H), 7.27-7.21 (m, 3H), 4.02 (s, 3H), 3.98 (s, 2H).

Process 2:

Step 4: Synthesis of 3-dimethylamino-1-(naphthalen-1-yl)propan-1-one oxalate

Paraformaldehyde (2.67 kg, 29.67 mol), dimethylamine hydrochloride (4.00 kg, 49.06 mol) was sequentially added to 1-naphthylethyl ketone (5.00 kg, 29.38 mol) of ethanol at 20-30 °C. 25 liters) of the solution, a catalytic amount of hydrochloric acid (139.23 g, 3.82 mol, 136.5 ml) was added, the reaction mixture was heated to 78-80 ° C reflux reaction for 48 hours, HPLC detection of 1-naphthyl ethyl ketone basic reaction is complete (content less than 5 %). The reaction solution was concentrated under reduced pressure to remove ethanol, then the residue was dissolved in water (25 liters) and extracted twice with ethyl acetate (5 liters x 2), and the aqueous phase was adjusted to pH with 1 mol/L aqueous sodium hydroxide至9至10, and then extracted twice with ethyl acetate (15 liters × 2), the second extracted organic phase was combined, washed with saturated brine (10 liters × 2), and concentrated under reduced Methylamino)-1-(1-naphthyl)propan-1-one (7.41 kg, yellow oil). The above crude product was dissolved in ethyl acetate (25 L), and then a solution of oxalic acid dihydrate (4.2 kg, 33.3 mol) in ethanol (20 L) was added dropwise to the above solution, and stirred at room temperature for 1 hour, with a large amount of white solid. Formed, then filtered, washed twice with ethyl acetate (5 L×2) to give 3-(dimethylamino)-1-(1-naphthyl)propan-1-one oxalate (6.5 kg, 22.18 moles, yield 56%, white solid). _{^{1 H NMR (400MHz, CDCl 3}} ) δ: 8.59 (d, J = 8.5Hz, 1H), 8.00 (d, J = 8.3Hz, 1H), 7.89 (d, J = 7.3Hz, 2H), 7.64-7.48 (m, 3H), 3.25 (t, J = 7.3 Hz, 2H), 2.83 (t, J = 7.3 Hz, 2H), 2.31 (s, 6H).

Step 5: Synthesis of 3-dimethylamino-1-(naphthalen-1-yl)propan-1-one

3-(Dimethylamino)-1-(naphthalen-1-yl)propyl-1-one oxalate (6.5 kg, 20.4 mol) was added to water (20 L) with stirring. The temperature was maintained at 0-10 ° C and the number of revolutions was 160 to 190 rpm to obtain a white suspension. A solution of cooled sodium hydroxide (1.8 kg, 45.0 moles) in water (10 liters) was slowly added to the suspension to maintain a temperature of 0 to 10 °C. It was observed that the white suspension gradually dissolved and a yellow oil formed. Stirring was continued for 1 hour after the addition of the sodium hydroxide solution. The reaction mixture was extracted with EtOAc (EtOAc)EtOAc. The organic phase was concentrated under reduced pressure to dryness to afford 3-(dimethylamino)-1-(naphthalen-1-yl)propyl-1-one (3.4 g,yield:yield: 70%). _{^{1 H NMR (400MHz, CDCl 3}} ) δ: 8.59 (d, J = 8.8Hz, 1H), 8.00 (d, J = 8.4Hz, 1H), 7.90 (d, J = 8.4Hz, 2H), 7.60-7.28 (m, 3H), 3.26 (t, J = 7.2 Hz, 2H), 2.83 (t, J = 7.2 Hz, 2H), 2.31 (s, 6H).

Process 3:

Step 5: 1-(5-(4-Chlorophenyl)-2-methoxypyridin-3-yl)-4-(dimethylamino)-2-(naphthalen-1-yl)-1-benzene Synthesis of butylbutan-2-ol

N-Butyllithium titration: Diphenylacetic acid (1.00 g, Alfa, 4.71 mmol) was added to tetrahydrofuran (10 ml) under a nitrogen atmosphere to give a colorless, transparent solution. A solution of butyllithium in n-hexane was slowly added dropwise to the above solution with a syringe. Observing the phenomenon, the solution partially yellowed during the addition process but disappeared rapidly. When a drop of a yellow solution is formed and the color does not fade within half a minute, the end point is recorded, and the volume of n-butyllithium (twice: 1.927 ml and 1.985 ml, respectively, the average volume is 1.95 ml) is recorded, so the n-butyl group used is used. The concentration of the lithium n-hexane solution was 2.42 mol/liter.

TMP (2.74 kg, 19.3 mol) was dissolved in anhydrous tetrahydrofuran (12 L), and the reaction temperature was cooled to -65 ° C in a dry ice acetone bath. n-butyllithium (8 L, 19.3 mol, 2.42 mol/L) was added dropwise. Alkane solution). The control temperature was between -20 ° C and -78 ° C. It was observed that the color of the reaction system gradually changed from light yellow to deep red to form a yellow suspension, and stirring was continued at this temperature for 30 minutes. Then reduce the reaction temperature to -75 ° C ~ -80 ° C began to slowly add 3-benzyl-5-(4-chlorophenyl)-2-methoxypyridine (4.08 kg, 12.9 mol) over 4-6 hours. An anhydrous tetrahydrofuran (6 liter) solution. Keeping the temperature between -65 ° C and -78 ° C, it can be observed that the exothermic color is dark red. After completion of the dropwise addition, 3-(dimethylamino)-1-(naphthalen-1-yl)propyl-1-one (3.26 kg, 12.9 mol, 90% purity) was slowly added dropwise over 2 to 4 hours. Anhydrous tetrahydrofuran (2.0 L) solution, the system exotherm is obvious, and the control flow rate is maintained at -65 ° C to -78 ° C. After the dropwise addition was completed, stirring was continued at -65 ° C to -78 ° C for half an hour. The content of 3-benzyl-5-(4-chlorophenyl)-2-methoxypyridine was determined by HPLC to be less than 10%, and the reaction solution was slowly added to a saturated ammonium chloride solution (40 liters) to be quenched. The liquid and aqueous phases were extracted with ethyl acetate (30 L). The organic phase was combined and washed with brine (30 ml). EtOAcjjjjjjjj 4) The mixed solvent was stirred at 5 ° C to 15 ° C for 16 hours to precipitate a white solid. Filtration, the filter cake was beaten with ethanol (4 liters × 2), filtered, and the filter cake was dried by vacuum drying to constant weight (50 ° C, 24-48 hours) to obtain the target compound 1-(5-(4-chlorophenyl)- 2-methoxypyridin-3-yl)-4-(dimethylamino)-2-(naphthalen-1-yl)-1-phenylbutan-2-ol (1.83 kg, yield 23.23%), As a white solid, HPLC determined the isomer A content to be 88.3% and the isomer B content to be 4.8%. _{^{1 H NMR (400MHz, CDCl 3}} ) δ: 8.85 (d, J = 2.3Hz, 1H), 8.64 (d, J = 8.7Hz, 1H), 8.32 (d, J = 2.4Hz, 1H), 7.98-7.86 (m, 2H), 7.72-7.61 (m, 2H), 7.57 (d, J = 8.4 Hz, 2H), 7.54-7.43 (m, 3H), 7.33 (t, J = 7.8 Hz, 1H), 7.20- 7.17 (m, 2H), 6.95-6.87 (m, 3H), 5.85 (s, 1H), 4.17 (s, 3H), 2.60-2.51 (m, 1H), 2.19-2.04 (m, 2H), 2.01- 1.97 (m, 7H).

Step 6: (1R,2S)-1-(5-(4-Chlorophenyl)-2-methoxypyridin-3-yl)-4-(dimethylamino)-2-(naphthalene-1- Synthesis of 1-phenylbutan-2-ol Compound I-2

method 1:

Two parallel batches were charged: R-(-)-binaphthol phosphate (519.3 g, 1.49 mol) was suspended in DMSO (1.0 L), heated to 50 ° C and stirred to dissolve until clear. 1-(5-(4-Chlorophenyl)-2-methoxypyridin-3-yl)-4-(dimethylamino)-2-(naphthalenemethanol-1-yl)-1-phenyl Butyl-2-ol (910 g, 1.49 mol, isomer A content of 88.3%) was added to a solution of ethanol (24 L) and added dropwise with stirring for 1 to 2 hours (196 rpm). A solution of R-(-)-binaphthol phosphate prepared above in DMSO (1.0 L). It was observed that the insoluble particulate compound began to dissolve, but a more viscous emulsion formed. After the addition, the reaction solution was further stirred at 15 to 35 ° C for 16 hours. The reaction solution was heated to reflux with an oil bath and reflux was continued for 1 hour. The heating was stopped and the reaction mixture was cooled to 15 to 35 ° C and stirring was continued for 16 hours. The reaction solution was filtered (two batches of combined treatment), the solid viscosity was large, the filtration was slow, and the filter cake was beaten three times with ethanol (20 liters). The combined organic layers were concentrated to a constant weight to give a yellow oil (5 kg). To this crude product was added water (10 liters) and ethyl acetate (5 liters), and the system was adjusted to pH with a 10% cold sodium hydroxide aqueous solution. 11. Stirring was continued for 1 hour, and then liquid separation was carried out, and a large amount of solid was precipitated in the system. The solid obtained by filtration was the isomer A (350 g, purity 97%, white solid). The filtrate was concentrated to a constant weight under reduced pressure at 50 ° C, and added with ethanol (1.0 L) at 15 - 35 ° C for 16 hours, filtered, and the filter cake was washed three times with ethanol (400 ml) to give a white solid which was dried in vacuo to dryness. Heavy (50 ° C, 24-48 hours) gave compound I-2 (400 g, purity 95%, ee value over 99.5%, yield 24%), white solid. _{^{1 H NMR (400MHz, CDCl 3}} ) δ: 8.85 (d, J = 2.3Hz, 1H), 8.64 (d, J = 8.7Hz, 1H), 8.32 (d, J = 2.4Hz, 1H), 7.98-7.86 (m, 2H), 7.72-7.61 (m, 2H), 7.57 (d, J = 8.4 Hz, 2H), 7.54-7.43 (m, 3H), 7.33 (t, J = 7.8 Hz, 1H), 7.20- 7.17 (m, 2H), 6.95-6.87 (m, 3H), 5.85 (s, 1H), 4.17 (s, 3H), 2.60-2.51 (m, 1H), 2.19-2.04 (m, 2H), 2.01- 1.97 (m, 7H).

Method 2:

S-binaphthol phosphate (473 g, 1.35 mol) was suspended in DMSO (1.1 L), and heated to 50 ° C to dissolve and clarify. 1-(5-(4-Chlorophenyl)-2-methoxypyridin-3-yl)-4-(dimethylamino)-2-(naphthalenemethanol-1-yl)-1-phenyl Butyl-2-ol (780 g, 1.35 moles, isomer A content 93.5%) was suspended in ethanol (22 L) solution. A solution of S-binaphthol phosphate in DMSO (1.1 L) prepared above was added dropwise over 1 hour with stirring (rpm 196 rpm). It was observed that the insoluble particulate compound began to dissolve, but a more viscous emulsion formed. After the addition, the reaction solution was further stirred at 15-35 ° C for 16 hours. The reaction was heated to reflux with an oil bath and reflux was continued for 1 hour. The reaction solution was stopped and cooled to 15 to 35 ° C and stirring was continued for 16 hours. The reaction solution was filtered, and the filtration was slow due to the large viscosity of the solid. The filter cake was beaten twice with ethanol (20 liters × 2) and filtered to give a white solid. The solid was mixed in water (3 liters) and ethyl acetate (3 liters), and a 10% aqueous sodium hydroxide solution was added to adjust the pH of the system. To 10-11, stirring was continued for 1 hour, liquid separation, organic phase was concentrated to constant weight, ethanol (1 L) was added and stirred for one hour, and filtered, and the obtained solid was recrystallized from ethanol (8 L) (heated to 80 ° C) Stirring was continued until clarification was cooled to 15-35 ° C and stirring was continued for 16 hours), and the mixture was cooled and filtered, and the obtained white solid was dried in vacuo to dryness (50 ° C, 24 to 48 hours) to give Compound I-2 (280 g, purity 99.2) %, ee value 99.1%, yield 38%, white solid). _{^{1 H NMR (400MHz, CDCl 3}} ) δ: 8.85 (d, J = 2.3Hz, 1H), 8.64 (d, J = 8.7Hz, 1H), 8.32 (d, J = 2.4Hz, 1H), 7.98-7.86 (m, 2H), 7.72-7.61 (m, 2H), 7.57 (d, J = 8.4 Hz, 2H), 7.54-7.43 (m, 3H), 7.33 (t, J = 7.8 Hz, 1H), 7.20- 7.17 (m, 2H), 6.95-6.87 (m, 3H), 5.85 (s, 1H), 4.17 (s, 3H), 2.60-2.51 (m, 1H), 2.19-2.04 (m, 2H), 2.01- 1.97 (m, 7H).

Step 7: (1R,2S)-1-(5-(4-Chlorophenyl)-2-methoxypyridin-3-yl)-4-(dimethylamino)-2-(naphthalenemethanol-1 -yl)-1-phenylbutyl-2-ol Fumarate Compound I-1

Compound I-2 (395 g, 0.735 mol) was added to a solution of fumaric acid (89.6 g, 0.772 mol) in isopropanol (7 L) over 1 to 2 hours, then activated charcoal (10 g) and isopropyl Alcohol (1.0 L). Heat to reflux and continue to reflux for 0.5 to 1 hour and the solids dissolved until clear. Hot filtration to obtain a colorless clear solution heated to reflux and kept at reflux for 1 hour, stop heating and slowly cool to 60 ° C, start solid precipitation, keep the temperature constant and continue to stir for 2 hours and then cool down to 15-35 ° C for 4-6 hours. Stirring was continued for 16 hours and a large amount of solid precipitated. The reaction solution was filtered, and the cake was dried by vacuum drying to constant weight (50 ° C, 24 to 48 hours) to obtain the title compound (1R, 2S)-1-(5-(4-chlorophenyl)-2-methoxy. Pyridin-3-yl)-4-(dimethylamino)-2-(naphthalenyl-1-yl)-1-phenylbutyl-2-ol fumarate, compound I-1 (398 g, The purity was 98.8%, the ee value was greater than 99.5%, the yield was 82.9%, white solid). ^{1 H NMR (400MHz, DMSO-} d 6) δ: 8.61 (d, J = 8.2Hz, 1H), 8.50 (br.s., 1H), 8.40 (br.s, 1H), 7.91 (br.s. , 2H), 7.71 (d, J = 8.0 Hz, 2H), 7.66 (d, J = 8.5 Hz, 2H), 7.56-7.54 (m, 3H), 7.35 (t, J = 7.7 Hz, 1H), 7.17 (d, J = 6.3 Hz, 2H), 6.92-6.80 (m, 3H), 6.53 (s, 2H), 5.73 (s, 1H), 4.13 (s, 3H), 2.37 (m, 1H), 2.13 ( m, 7H), 2.07-1.97 (m, 1H), 1.90 (d, J = 8.9 Hz, 1H).

Example 2: Preparation of Form I of Compound I-1

365 g of the compound I-1 obtained in the above step 7 was suspended in a mixed solvent of acetone/methanol (20/1, 730 ml), and stirred at 15 to 35 ° C for 16 to 20 hours, filtered, and the filter cake was vacuum dried. Drying to constant weight (50 ° C, 24-48 hours) gave Form I of Compound I-1 (265 g, purity 99.7%, ee value greater than 99.5%, yield 72.6%, white solid).

Example 3: Preparation of Form II of Compound I-1

About 50 mg of Form I of Compound I-1 was taken, and 0.1 mL of tetrahydrofuran was added to form a suspension. The suspension sample was shaken on a constant temperature uniform (40 ° C) for 2 days (protected from light). The residual solid was centrifuged and dried in a vacuum oven at 40 ° C overnight to obtain compound I-1. Form II.

Example 4: Preparation of Form III of Compound I-1

The preparation of Form III was the same as Form II, and only the solvent tetrahydrofuran was changed to 0.2 mL of acetone-water (2:1).

Example 5: Preparation of Compound 2

Step 11:1-(3,5-Dichlorophenyl)-3-(dimethylamino)propan-1-one

Mix 3,5-dichloroacetophenone (25.0 g, 132.25 mmol), dimethylamine hydrochloride (43.13 g, 529 mmol) and paraformaldehyde (15.49 g, 171.93 mmol) in ethanol at room temperature Concentrated hydrochloric acid (4.73 ml) was added (300 ml), the reaction mixture was heated to 80 ° C and stirring was continued for 48 to 52 hours. The reaction mixture was evaporated to dryness crystals crystals crystals crystals The aqueous phase was adjusted to pH -12 with a 10% aqueous sodium hydroxide solution, and then extracted with dichloromethane (200 ml × 3). The organic phase extracted twice and dried over anhydrous sodium sulfate. The title compound 1-(3,5-dichlorophenyl)-3-(dimethylamino)propan-1-one (24 g, purity 83%) was obtained as a yellow oily liquid.

Step 12: 1-(5-(4-Chlorophenyl)-2-pyridyl)-2-(3,5-dichlorophenyl)-4-(dimethylamino)-1-phenyl-butyl Alkanol-2-ol

TMP (16.41 g, 116.2 mmol) was dissolved in tetrahydrofuran (200 ml) with stirring at room temperature, replaced with nitrogen, cooled to -78 ° C with a dry ice-acetone bath, and n-butyllithium (46.48 ml, 116.2 m) was slowly added dropwise. Molar, 2.5 mol/L n-hexane solution), stirring was continued for 30 minutes while maintaining the temperature at -78 to -20 °C. The temperature of the reaction system was lowered to -78 to -65 ° C, and 3-phenyl-5-(4-chlorophenyl)-2-methoxy-pyridine (24.0 g, 77.4 mmol) of tetrahydrofuran (200 ml) was added. The solution was stirred for 10 minutes after completion of the dropwise addition, and then 1-(3,5-dichlorophenyl)-3-(dimethylamino)propan-1-one (23.84 g, 77.47 mmol) of tetrahydrofuran (23. The solution was slowly added dropwise to the reaction system, and the reaction solution was further stirred at -78 to -65 ° C for 20 minutes. The mixture was quenched with EtOAc EtOAc (EtOAc m. The crude product was obtained. The title compound (1R,2S)-1-(5-(4-chlorophenyl)-2-pyridyl)- 2-(3,5-Dichlorophenyl)-4-(dimethylamino)-1-phenyl-butan-2-ol and (1S,2R)-1-(5-(4-chlorophenyl) a mixture of 2-pyridyl)-2-(3,5-dichlorophenyl)-4-(dimethylamino)-1-phenyl-butan-2-ol (5.0 g, purity 99%) , yield 11.5%, white solid). _{^{1 H NMR (400MHz, CDCl 3}} ) δ: 8.66 (d, J = 2.51Hz, 1H), 8.39 (br.s., 1H), 8.25 (d, J = 2.51Hz, 1H), 7.50-7.48 (m , 2H), 7.42-7.40 (m, 2H), 7.40-7.30 (m, 4H), 7.11-7.02 (m, 4H), 4.76 (s, 1H), 4.06 (s, 3H), 2.26-2.23 (m , 1H), 2.12-2.04 (m, 8H), 1.73-1.70 (m, 1H).

Step 13: (1R,2S)-1-(5-(4-Chlorophenyl)-2-pyridyl)-2-(3,5-dichlorophenyl)-4-(dimethylamino)- 1-phenyl-butan-2-ol

The corresponding isomer (1R,2S)-1-(5-(4-chlorophenyl)-2-pyridyl)-2-(3,5-difluorobenzene) obtained in Step 10 at 0 °C (4-(dimethylamino)-1-phenyl-butan-2-ol and (1S,2R)-1-(5-(4-chlorophenyl)-2-pyridyl)-2- (3,5-Difluorophenyl)-4-(dimethylamino)-1-phenyl-butan-2-ol (8.0 g, 14.39 mmol) was suspended in ethanol (240 mL) and then A solution of R-(-)-binaphthol phosphate (5.01 g, 14.39 mmol) in dimethyl sulfoxide (24 ml) was slowly added dropwise to the substrate solution, and the reaction mixture was stirred at 20 ° C for 8 hours. The reaction solution was then heated to 80 ° C and stirred for 1 hour, slowly cooled (2 to 4 hours) to 20 ° C and stirring was continued for 16 hours. The reaction solution was filtered, and the filtrate was concentrated under reduced pressure, and then dissolved in ethyl acetate (40 ml), and adjusted to basic (pH: 12) with 10% sodium hydroxide, partitioned, and the organic phase was dried over anhydrous sodium sulfate Dry and concentrate to give the crude product. The crude product was added to ethanol (100 ml), stirred at 20 to 30 ° C for 1 to 2 hours, filtered, and the filter cake was dried by vacuum drying to constant weight (50 ° C, 24 to 48 hours) to obtain the desired product (1R, 2S)-1-(5-(4-chlorophenyl)-2-pyridyl)-2-(3,5-dichlorophenyl)-4-(dimethylamino)-1-phenyl-butyl Alkan-2-ol (3.1 g, purity 98.9%, ee value 98.1%, yield 38.8%, white solid). _{^{1 H NMR (400MHz, CDCl 3}} ) δ: 8.66 (d, J = 2.51Hz, 1H), 8.39 (br.s., 1H), 8.25 (d, J = 2.51Hz, 1H), 7.50-7.48 (m , 2H), 7.42-7.40 (m, 2H), 7.40-7.30 (m, 4H), 7.11-7.02 (m, 4H), 4.76 (s, 1H), 4.06 (s, 3H), 2.26-2.23 (m , 1H), 2.12-2.04 (m, 8H), 1.73-1.70 (m, 1H).

Example: Stability test of 6:I crystal form in non-solvent

Take appropriate amounts of multiple forms of Form I, add 0.1-0.3 mL of the single or mixed solvent in the table below, stir at 40 ° C for 2 days and centrifuge. The solids in all samples were collected, dried overnight in a vacuum oven (40 ° C), and XRPD was tested for crystal form. The results are shown in Table-4.

Table-4 Stability Test of Form I in Solvent Free

Serial number	Solvent	Appearance (2 days)	result
1	Methanol	Suspension	I crystal form
2	Ethanol	Suspension	I crystal form
3	acetone	Suspension	I crystal form
4	Ethyl acetate	Suspension	I crystal form
5	Tetrahydrofuran	Suspension	I crystal form
6	Methanol-water (3:1)	Suspension	I crystal form
7	Ethanol-water (3:1)	Suspension	I crystal form / III crystal form
8	Acetone-water (2:1)	Suspension	III crystal form

Example 7: Solid stability test of crystal form I under high temperature, high humidity and strong light conditions

Approximately 10 mg of the I crystal form sample was weighed and placed on the bottom of the glass sample vial to form a thin layer. The samples placed at 60 ° C and 92.5% RH were sealed with aluminum foil paper, and small holes were placed on the aluminum foil to ensure that the sample could be in full contact with ambient air; the sample placed under strong light (5Klux) was used for the screw bottle. The lid is sealed. Samples placed under different conditions were sampled on day 5 and 10 days. The test results were compared with the initial test results on day 0. The test results are shown in Table-5 below:

Table-5 Solid Stability Test of Form I

Pharmacological part

Part I: Testing the anti-tuberculosis mycobacterial compound in vitro efficacy using M. smegmatis strain ATCC19420

On the day of the test, the compound was dissolved in pure DMSO (Sigma 276855-2L) to a concentration of 12.8 mg/ml as a mother liquor of the compound. 30 μl of DMSO was added to all wells of a v-bottom 96-well plate (Axygen-wipp 02280). Add 30 μl of the compound mother solution to the well of the first column, mix by pipetting, and add 30 μl from the first column of wells to the second column of wells and mix by pipetting. Take this to the 11th column. Column 12 was not administered and contained only 30 μl of DMSO. This is the compound "motherboard". From column 1 to column 12, the corresponding compound concentrations were 6.4, 3.2, 1.6, 0.8, 0.4, 0.2, 0.1, 0.05, 0.025, 0.0125, 0.00625, 0 mg/ml. For compounds with good efficacy, reduce the test concentration appropriately. A u-bottom 96-well plate (Costar 3788) was used as the "sub-plate". 98 μl of CA-MHB (BD-212322) medium containing 0.02% Tween 80 was added to the wells of all daughter plates. 2 μl of compound was pipetted from the mother plate into the daughter plate at the corresponding position.

The bacteria were inoculated on Roche modified slant medium (Difco-244420) two days in advance, and cultured in a 37 ° C incubator for 48 hours. On the day of the test, bacterial colonies were collected from the slant of the medium and suspended in sterile physiological saline containing 0.02% Tween 80. Add 7-10 sterile glass beads of 3 mm diameter to the bacterial solution and use a vortex to break up the bacteria at maximum speed. The turbidity of the bacterial solution was adjusted to 0.10 using a Siemens MicroScan turbidity meter, and the corresponding bacterial concentration was -1.5 x 10 ⁸ cfu/ml. The bacterial solution was diluted 20 times with CA-MHB + 0.02% Tween 80 medium, and then diluted 25 times (500 times total). The diluted bacterial solution will be used to inoculate the daughterboard.

100 μl of the bacterial solution was added to each well of the daughter plate. Each well will contain: ~ 3.0 x 10 ⁴ cfu bacteria, 1% DMSO, and a gradient dilution of the compound in 200 μl CA-MHB + 0.02% Tween 80 medium. The finished daughterboard was placed in a 30 ° C incubator for cultivation. The minimum inhibitory concentration (MIC) was read after 72 hours.

The standard for reading MICs is defined by the CLSI method M7-A7 as the lowest concentration of a drug that completely or significantly inhibits bacterial growth. Compound The test results are listed in Table-6.

Part II: Testing the anti-tuberculous mycobacterial compound in vitro efficacy using H37Rv strain

On the day of the test, the compound was dissolved in pure DMSO (Sigma 276855-2L) to a concentration of 10 mg/ml as a mother liquor of the compound. 30 μl of DMSO was added to the wells of columns 2 to 11 of the v-bottom 96-well plate (Axygen-wipp 02280). Add 30 μl of compound mother liquor to the wells of column 2, mix well, and take 30 μl from the second column of wells and add to the third column of wells and mix by pipetting. Take this to the 10th column. Column 11 was not administered and contained only 30 μl of DMSO. This is the compound "motherboard". From column 2 to column 11, the corresponding compound concentrations were 5, 2.5, 1.25, 0.625, 0.3125, 0.156, 0.078, 0.039, 0.02, 0 mg/ml. For compounds with good efficacy, reduce the test concentration appropriately. A flat-bottom 96-well plate (Greiner 655090) was used as the "sub-plate". 98 μl of 7H9 (Sigma M0178) medium was added to the wells of all daughter plates. 2 μl of compound was pipetted from the mother plate into the daughter plate at the corresponding position. Subplates A and H, columns 1 and 12 contain only 7H9 medium.

The H37Rv strain in the glycerol cryotube was inoculated into 7H9 medium containing 0.05% Tween 80, and cultured at 37 ° C for 4 weeks in a shaker at 200 rpm. The bacterial solution was washed twice with 7H9 medium containing 0.05% Tween 80 and resuspended in the same medium. The absorbance of the bacterial solution was adjusted to an OD ₅₅₀ = 0.4-0.5 using the same medium. This bacterial solution was dispensed into a microcentrifuge tube and stored at -80 °C. Storage time is less than 1 month. On the day of the test, the dispensed bacteria were lyophilized. The lyophilized broth was diluted 20-fold with 7H9 medium and then diluted 50-fold, and diluted 1000 times. This broth was used to inoculate the daughter plate. 100 μl of the bacterial solution was inoculated into each well of the daughter plate, and the column 12 was added with 100 μl of 7H9 medium without adding a bacterial solution.

The test panels were placed in a 37 ° C incubator and the humidity was maintained at >80%. Starting one week later, add 12.5 μl of 7H9 medium containing 20 % Tween 80 and 20 μl of Alamar Blue (Invitrogen DAL1100) to a column of bacteria-containing wells and a column 12 containing no bacteria. Observed after 24 hours of culture. When the bacterial solution in the first column of pores can reduce the added Alamar blue to pink within 24 hours, add 7H9 medium containing 20% Tween 80 and Alamar blue to all wells on the test plate, 37 ° C Fluorescence values were measured after 24 hours of continued incubation.

The minimum inhibitory concentration (MIC) is defined as the minimum drug concentration that can completely inhibit the discoloration of Alamar blue by visual observation, or the minimum drug concentration that can inhibit the formation of more than 90% reduced Alamar blue by fluorometer. The compound test results are shown in Table-6.

Table-6 In vitro screening results

Note: ATCC--American type culture strains; MABA--microplate Alamar blue color test; LORA--recovery test under anaerobic conditions; Vero Cell--African green monkey kidney cells; IC50--half suppression Concentration; Hela--human cervical cancer cells; CC50--half cytotoxic concentration.

Analysis of results: Compound I-2 has a good inhibitory activity against Mycobacterium smegmatis, and compound I-2 is still in aerobic (MABA) The inhibitory activity against Mycobacterium tuberculosis under anaerobic (LORA) conditions is superior to or equivalent to the marketed anti-tuberculosis drug betaxazoline. Moreover, Compound 1-2 showed no significant cytotoxicity against both Vero and Hela cells.

Part III: Evaluation of in vitro efficacy of compounds against drug-resistant Mycobacterium tuberculosis

Using the same method as mentioned in the second section, we tested the activity of Compound I-2 with a drug-sensitive and drug-resistant M. tuberculosis strain. The results are shown in Table-7.

Table-7: MIC (uM) of some test compounds against drug-sensitive and drug-resistant Mycobacterium tuberculosis

Note: MIC--minimum inhibitory concentration; MABA--microplate Alamar blue color test; vs--pair, relative; H37Rv--wild type H37Rv strain; rRMP-- rifampicin-resistant tuberculosis Bacillus strain; rINH - a strain of M. tuberculosis resistant to isoniazid.

Analysis of the results: Compound I-2 not only inhibited the wild-type Mycobacterium tuberculosis H37Rv, but also showed good inhibition to rifampicin and isoniazid-resistant strains. The inhibitory activities of the three strains tested were comparable to those of the marketed antitubercular drug betaxazoline.

Claims (10)

1. a method for preparing a compound of formula (I),

   

   It contains the following steps:

   

   among them,

   R ^{1 is} selected from a 6- to 12-membered aryl group, a 6- to 12-membered heteroaryl group, a 6- to 12-membered aryl-alkylene group, and a 6 to 12-membered heterocyclic group, which are optionally substituted by 0, 1, 2 or 3 R ⁰¹ . Aryl-alkylene;

   HX is selected from an organic or inorganic acid;

   The base A is selected from the group consisting of an alkali metal base, an alkaline earth metal base or an organometallic base;

   The molar ratio of the compound (II) to the base A is 1:1 to 5;

   The molar ratio of the compound (II) to the compound (III) is 1:1 to 2;

   The reaction solvent is selected from a single ether solvent or a mixed solvent of several ether solvents;

   The amount of the reaction solvent is 3 to 20 times the weight of the compound (IV);

   The reaction temperature is -80 to 0 ° C;

   The reaction time is 1 to 24 hours;

   R ^{01 is} selected from the group consisting of F, Cl, Br, I, CN, OH, CH(CH ₃ ) ₂ , C(CH ₃ ) ₃ , N(CH ₃ ) ₂ , NH(CH ₃ ), NH ₂ , CHO, COOH, C(=O)NH ₂ , S(=O)NH ₂ , S(=O) ₂ NH ₂ , CF ₃ , CF ₃ O, (NH ₂ )CH ₂ , (HO)CH ₂ , CH ₃ C(= O), CH ₃ OC(=O), CH ₃ S(=O) ₂ , CH ₃ S(=O); the "hetero" represents a hetero atom selected from N, O or S;

   The number of heteroatoms is independently selected from 1, 2 or 3, respectively.
2. The production method according to claim 1, wherein R ¹ is a naphthyl group or a phenyl group optionally substituted by 0, 1, 2 or 3 R ⁰¹ ;

   Specifically, R ^{1 is} selected from

   
3. The production method according to claim 1, wherein the alkali metal base is selected from the group consisting of lithium hydroxide, sodium hydroxide, potassium hydroxide, barium hydroxide, sodium carbonate, potassium carbonate, cesium carbonate, sodium hydrogencarbonate and/or carbonic acid. Potassium hydrogen

   The alkaline earth metal base is selected from the group consisting of sodium hydride, potassium hydride and/or calcium hydride;

   The organometallic base is selected from the group consisting of n-butyl lithium, lithium diisopropylamide, lithium 2,2,6,6-tetramethylpiperidine, lithium bis(trimethylsilyl)amide, sodium methoxide, lithium t-butoxide, Sodium tert-butoxide, potassium t-butoxide, sodium ethoxide and/or aluminum isopropoxide.
4. The preparation method according to claim 1, wherein the molar ratio of the compound (II) to the base A is 1:1.2 to 2;

   The reaction temperature is -80 to -60 ° C;

   The reaction time is 2 to 12 hours;

   Specifically, the reaction time is 4 to 8 hours;

   The reaction solvent is selected from the group consisting of tetrahydrofuran, diethyl ether and/or isopropyl ether; and/or

   The reaction solvent is used in an amount of 5 to 10 times by weight based on the weight of the compound (IV).
5. The preparation method according to any one of claims 1 to 4, which comprises the following reaction route:

   

   

   among them,

   The base B is selected from the group consisting of lithium hydroxide, sodium hydroxide, potassium hydroxide, barium hydroxide, sodium carbonate, potassium carbonate, barium carbonate, sodium hydrogencarbonate, potassium hydrogencarbonate, sodium methoxide, lithium t-butoxide, sodium t-butoxide, Potassium tert-butoxide, sodium ethoxide or aluminum isopropoxide;

   The chiral acid is selected from the group consisting of α-hydroxypropionic acid, α-hydroxysuccinic acid, α,β-dihydroxysuccinic acid, α-hydroxyphenylacetic acid, β-hydroxy acid, and compound (VI);

   

   n is 0, 1 or 2;

   R ² and R ⁴ are each independently selected from H, F, Cl, Br, I, or selected from, optionally substituted by 0, 1, 2 or 3 R ⁰¹ : C _1-8 alkoxy, C _{1- 8-} alkyl, Si(Ph) ₃ , 6- to 12-membered aryl;

   R ³ and R ⁵ are each independently selected from H, F, Cl, Br, I, NO ₂ , OH, or selected from C _1-8 alkoxy optionally substituted by 0, 1, 2 or 3 R ⁰¹ a group, a C _1-8 alkyl group, a 6 to 12 membered aryl group;

   Optionally, position 13 and position 14 is substituted with R ³ or at position 14 and position 15 is substituted with R ³ may be joined together to form a 6 to 12 membered aryl ring;

   Optionally, the 8 position and R ⁵ is substituted at position 9 or position 9 and joined together at position 10 substituted with R ^5, form a 6-12 membered aryl ring;

   specifically,

   R ³ and R ⁵ are each independently selected from H;

   R ^{2 is} substituted at 2 positions;

   R ^{4 is} substituted at 6 positions;

   R ² and R ⁴ are each independently selected from: H, Si(Ph) ₃ ,

   

   The solvent for preparing the compound (V) from the compound (IV) is selected from the group consisting of acetone, methyl ethyl ketone, ethanol, methanol, isopropanol, tert-butanol, ethyl acetate, t-butyl acetate, DMF, DMSO, DMA and / or NMP, a single solvent or a mixed solvent of several solvents;

   The molar ratio of the chiral acid to the compound (IV) is from 0.5 to 1.5;

   Specifically, the molar ratio of the chiral acid to the compound (IV) is from 0.8 to 1.2;

   More specifically, the molar ratio of the chiral acid to the compound (IV) is 1.0;

   HX is selected from the group consisting of hydrochloric acid, sulfuric acid, phosphoric acid, oxalic acid, citric acid, maleic acid or fumaric acid.
6. The production method according to claim 5, wherein the compound (VI) is selected from the group consisting of

   

   
7. As a compound of the formula wherein the intermediate of the compound of formula (I) is prepared:

   
8. Compound I-1 represented by the following formula,

   
9. The X-form, Form II and Form III of Compound I-1, the XRPD patterns thereof are shown in Figures 1, 4 and 7, respectively.
10. The method for producing a crystalline form of the formula I according to claim 9, which comprises preparing a compound I-1 of any one form by adding it to a solvent, wherein

    The solvent is selected from the group consisting of an alcohol, a ketone solvent, or a mixed solvent of an alcohol solvent and a ketone solvent;

    The solvent is used in an amount of from 3 to 50 times the weight of the compound I-1;

    specifically,

    The alcohol solvent is selected from the group consisting of methanol, ethanol, isopropanol and/or n-butanol;

    The ketone solvent is selected from the group consisting of acetone and/or methyl ethyl ketone;

    The mixed solvent is a mixed solvent of methanol and acetone; or

    The volume ratio of the mixed solvent of methanol and acetone is 1:5 to 30.

Images