EA045390B1 - METHOD FOR SYNTHESIS OF FUROIMIDAZOPYRIDINE COMPOUND, CRYSTAL FORM OF FUROIMIDAZOPYRIDINE COMPOUND AND CRYSTAL FORM OF ITS SALT - Google Patents

Description

Field of technology

The present invention relates to the field of drug synthesis, in particular to a method for the synthesis of the compound 2-[(2R,5S)-5-[2-[(R)-1-hydroxyethyl]furo[3,2-b]imidazo[4, 5-d]pyridin-1-yl]tetrahydropyran-2-yl]acetonitrile (hereinafter referred to as compound I) as a selective inhibitor of JAK1/TYK2 kinase. The present invention also relates to the crystalline form of compound I and its salts, as well as methods for their preparation. In addition, the present invention also relates to pharmaceutical compositions and pharmaceutical compositions containing the specified crystalline form of compound I and/or the crystalline form of its salts, as well as the use of the specified crystalline form of compound I and its salts for the treatment of diseases and conditions associated with JAK1/ TYK2.

State of the art

Protein kinases are a family of proteins that play important roles in modulating a variety of cellular processes and maintaining cellular functions. These kinases contain at least a non-receptor tyrosine kinase belonging, for example, to the Janus kinase family (JAK1, JAK2, JAK3 and TYK2); a receptor tyrosine kinase such as platelet-derived growth factor receptor (PDGFR), and a serine/threonine kinase such as b-RAF.

The Janus kinase family includes 4 known members: JAK1, JAK2, JAK3, and tyrosine kinase 2 (TYK2). These cytoplasmic tyrosine kinases are cytokine membrane receptors (eg, common γ-chain receptors and transmembrane glycoprotein 130 (gp130)) (Murray, J. Immunol., 178(5):2623-2629, 2007). Nearly 40 cytokine receptors signal through a combination of these four members of the Janus kinase family and their 7 downstream substrates: members of the signal transduction activator of transcription (STAT) family (Ghoreschi et al., Immunol Rev., 228(I):273- 287, 2009). A cytokine that binds to its receptor activates Janus kinases by trans- and/or autophosphorylation. Activated JAK family kinase phosphorylates cytokine receptor residues, creates binding sites for proteins (such as STAT factor and other regulators) containing Src homology 2 (SH2), and activates them. Activated STAT enters the cell nucleus and begins to stimulate the expression of survival factors, cytokines, chemokines and leukocyte transport molecules (Schindler et al., J. Biol. Chem., 282(28):20059-20063, 2007). Activation of JAK kinases also induces cell proliferation through phosphoinositide 3-kinase (PI3K) and protein kinase B-mediated pathways.

The kinases JAK3 and JAK1 are components of the cytokine receptor-common γ-chain junction, and blocking either of them can inhibit inflammatory cytokine signaling (interleukins (IL)-2, 4, 7, 9, 15, and 21) (Ghoreschi et al., Immunol. Rev., 228(1):273-287, 2009). However, other significant cytokines (such as IL-6) depend only on JAK1 in terms of their contribution to pathology. Consequently, blocking JAK1 inhibits the signaling of many proinflammatory cytokines (Guschin et al., EMBO J., 14(7):1421-1429, 1995). Clinical efficacy of an IL-6 receptor neutralizing antibody, tocilizumab, has been reported in rheumatoid arthritis (RA) (Maini et al., Arthritis Rheum., 54(9):2817-2829, 2006).

International application WO 2018067422 A1 discloses 1H-furo[3,2-b]imidazo[4,5d]pyridine derivatives as selective JAK1 kinase inhibitors and methods for their preparation, and discloses compound I and a method for its preparation. The synthesis scheme is as follows:

O

Compound I

Biological testing shows that Compound I is a potent and selective inhibitor of JAK1, exhibits selective inhibition of IL-6-induced STAT3 phosphorylation, and does not exhibit selective inhibition of thrombopoietin-induced STAT3 phosphorylation. However, the international application WO 2018067422 A1 does not demonstrate the biological activity of TYK2. In addition, the disclosed method for preparing Compound I involves the use of high temperature, produces too many impurities and provides low

- 1 045390 product yield, as a result of which this method is not suitable for large-scale production.

Therefore, it is necessary to develop a method for preparing compound I under milder reaction conditions, with higher yields of higher purity, and also suitable for large-scale/industrial production.

Until now, the crystalline form of compound I and its salt has not been reported in the prior art. When developing and researching a drug, it is necessary to comprehensively and systematically screen polymorphisms and select the crystalline form most suitable for the development being carried out. Accordingly, it is necessary to further screen the crystalline forms of compound I and its salts, identifying crystalline forms with good stability and low hygroscopicity, suitable for large-scale production and providing greater and improved opportunities for subsequent drug development.

The essence of the invention

The object of the present invention is to provide a process for the preparation of a compound of formula I (ie compound I) under mild reaction conditions, with high yield and high purity of the product, suitable for industrial production. The synthesis scheme of this method is as follows:

I

This method includes the following stages:

step 1, including adding ethanol, a compound of formula IV, a compound of formula V and DIPEA (diisopropylethylamine) to the reaction vessel, stirring;

heating to a temperature of 65-90°C, maintaining the specified temperature and stirring the reaction mixture overnight;

completion of the reaction and reduction of the temperature in the system to 15-30°C;

adding water dropwise to the system followed by stirring;

filtering and washing the filter cake;

drying the filter cake to obtain a compound of formula III;

step 2, comprising adding tetrahydrofuran (THF), the compound of formula III obtained in step 1, and palladium-carbon to the reaction vessel;

purging the system with nitrogen and then hydrogen;

maintaining a temperature of 20-35°C and stirring for 16-120 hours at a hydrogen pressure of 0.1-1.0 MPa;

after completion of the reaction, filtering the reaction liquid and washing the filter cake;

combining the filtrates and concentrating to obtain a concentrate of the compound of formula II;

step 3, including adding THF, (R)-lactamide and Et ₃ O-BF4 to the first reaction vessel, stirring, dissolving for subsequent use;

adding said concentrate of the compound of formula II and ethanol to a second reaction vessel, heating the materials in the second vessel to a temperature of 40-85°C;

adding materials from the first reaction vessel dropwise to the second reaction vessel and upon completion of addition maintaining the temperature at 40-85°C to allow the mixture to react

- 2,045,390 materials in the second reaction vessel for 0.5-6 hours;

after completion of the reaction, bringing the pH of the system to 1-3, extraction with an organic solvent, removing the organic phase, bringing the pH of the aqueous phase to 9-10 with an aqueous solution of inorganic alkali, filtration, drying the filter cake to obtain a compound of formula I.

In a preferred embodiment of the present invention, in said step 1, the volume/mass ratio (ml/g) of ethanol to the compound of formula IV is from 5:1 to 20:1, preferably 10:1;

the molar ratio of the compound of formula IV, the compound of formula V and DIPEA is 1:1-1.1:2-3, preferably 1:1.01:2.2;

after turning on stirring in a nitrogen atmosphere, the reaction mixture is heated to 65-90°C, preferably to 70-90°C, more preferably to 70-80°C, maintain the specified temperature and stir for 5-16 hours, preferably for 10- 16 hours;

after completion of the reaction, reduce the temperature in the system to 15-25°C;

the volume-mass ratio (ml/g) of water added to the system and the compound of formula IV is from 10:1 to 20:1, preferably 15:1;

after adding water to the system, stirring is carried out for 2-6 hours, preferably for 4 hours, at a temperature of 0-30°C, preferably at 5-15°C, more preferably at 5-10°C;

the filter cake is washed with a solution of ethanol in water, the volume ratio (ml/ml) of ethanol and water in the solution being from 1:1 to 1:2, preferably from 1:1.5 to 1:2;

the volume-mass ratio (ml/g) of the ethanol solution and the compound of formula IV is from 2:1 to 10:1, preferably from 2:1 to 5:1, more preferably from 2:1 to 3:1;

The filter cake is dried under vacuum or under a blower at a temperature of 45 to 55°C, preferably at 50°C.

In a preferred embodiment of the present invention, in said step 2, the volume/mass ratio (ml/g) of THF to the compound of formula III is from 10:1 to 70:1, preferably from 20:1 to 70:1;

palladium on carbon is 5% Pd/C, palladium on carbon with 50% moisture, the weight ratio of palladium on carbon to the compound of formula III is from 0.15:1 to 0.16:1, preferably 0.15:1;

maintain the temperature at 25-35°C and stir for 24-96 hours at a hydrogen pressure of 0.5-1.0 MPa;

the filter cake is washed with THF, wherein the concentrate of the compound of formula II obtained by combining the filtrates and concentrating is the compound of formula II in THF, the volume-mass ratio (ml/g) of THF for washing and the compound of formula II is from 2:1 to 4: 1, preferably from 2:1 to 3:1 (the weight of the compound of formula II was determined based on 100% yield in step 2);

Preferably, the THF in the solution of the compound of formula II is replaced by ethanol to obtain a solution of the compound of formula II in ethanol, wherein the volume/weight ratio (ml/g) of ethanol to the compound of formula II is from 2:1 to 5:1, preferably from 2:1 to 4 :1, more preferably from 2:1 to 3:1 (the weight of the compound of formula II was determined based on 100% yield in step 2).

In a preferred embodiment of the present invention, in said step 3, the volume/mass ratio (ml/g) of THF to concentrate of the compound of formula II is from 6:1 to 12:1;

the molar ratio of the concentrate of the compound of formula II, (R)-lactamide and Et ₃ O-BF ₄ is 1:4-5:4-5;

the volume-mass ratio (ml/g) of ethanol and concentrate of the compound of formula II is from 10:1 to 16:1, preferably 14:1;

after adding the concentrate of the compound of formula II and ethanol to the second reaction vessel, heat the materials in the second reaction vessel under a nitrogen atmosphere to a temperature of 40-85°C, preferably 45-70°C, more preferably 45-50°C;

maintain the temperature at 45-70°C, preferably 45-50°C, allowing the materials mixed in the second reaction vessel to react for 2-5 hours, preferably for 3 hours;

after completion of the reaction, adjust the pH of the system to 1-3 with hydrochloric acid, which is a 1 M or 12 M HCl solution, preferably 12 M HCl;

said aqueous inorganic alkali solution is a saturated sodium carbonate solution or a saturated potassium carbonate solution, preferably a saturated potassium carbonate solution;

the organic solvent used to extract the product after completion of the reaction is dichloromethane or ethyl acetate;

The precipitate obtained on the filter is dried under vacuum or blower at a temperature of 50-55°C.

In a preferred embodiment of the present invention, the compound of formula I obtained in said step 3 is separated and purified by column chromatography, where as

- 3 045390 eluent using a mixture of solvents, namely ethyl acetate and ethanol (V _EA :V _EtOH =100:1-30:1, ml/ml).

Another object of the present invention is to provide a crystalline form of a compound of formula I, hereinafter referred to as crystalline form 1 of a compound of formula I.

The X-ray powder diffraction pattern of crystalline form 1 of the compound of formula I according to the present invention contains characteristic peaks at 2θ angles of 8.5±0.2°, 14.8±0.2° and 16.1±0.2°.

In a preferred embodiment of the present invention, the X-ray powder diffraction pattern of crystalline Form 1 of the compound of formula I contains characteristic peaks at 2θ angles of 8.5±0.2°, 14.8±0.2°, 16.1±0.2° , 17.1±0.2°, 18.8±0.2° and 19.6±0.2°.

In yet another preferred embodiment of the present invention, the X-ray powder diffraction pattern of crystalline form 1 of the compound of formula I contains characteristic peaks at 2θ angles of 8.5±0.2°, 14.8±0.2°, 16.1±0, 2°, 17.1±0.2°, 18.8±0.2°, 19.6±0.2°, 23.8±0.2°, 25.3±0.2° and 26, 1±0.2°.

X-ray powder diffraction data for crystalline form 1 of the compound of formula I are presented without any restrictive interpretation in table. 1.

Table 1

Diffraction angle 20 Quantities a d Relative intensity, % (Calculated according to peak height) 8.5 10.3378 100 14.8 5.9874 15.7 16.1 5.5068 16.1 17.1 5.1787 11.5 18.8 4.7149 17.9 19.6 4.5237 26.1 23.8 3.7351 19.3 25.3 3,519 24.1 26.1 3.4072 16.1

Without being limited to anything, an X-ray powder diffraction (XRPD) diffraction pattern of crystalline Form 1 of the compound of Formula I according to the present invention is shown in FIG. 1.

Without being limited to anything, a differential scanning calorimetry (DSC) thermogram of crystalline Form 1 of the compound of Formula I according to the present invention is shown in FIG. 2A. The DSC thermogram shows that the initial melting point of crystalline form 1 of the compound of formula I according to the present invention is 160.76° C., and the broad endothermic peak at 91.85° C. represents an anhydrous solvent peak.

Without being limited to anything, a thermogravimetric analysis (TGA) thermogram of crystalline Form 1 of the compound of Formula I according to the present invention is shown in FIG. 3. The TGA thermogram shows that the mass loss effect of 5.353% for crystalline form 1 of the compound of formula I according to the present invention in the temperature range from 25 to 133° C. corresponds to the mass percent loss of one water molecule.

Without being limited to anything, a graph of the dynamic vapor sorption isotherm (DVA) of crystalline Form 1 of the compound of Formula I according to the present invention is shown in FIG. 4. The DSP isotherm plot shows that a 5.2% increase in mass for crystalline form 1 of the compound of formula I according to the present invention due to moisture absorption when changing relative humidity in the range from 0 to 95% indicates that the sample is hygroscopic, but water cannot be completely removed from it during desorption (2% remains).

The present invention provides a method for preparing crystalline Form I of a compound of formula I. Specifically, the method is as follows.

Method 1: Dissolving the compound of Formula I in a solvent, stirring at room temperature, then adding water to the resulting solution of the compound of Formula I, stirring, filtering and drying to obtain crystalline Form I of the compound of Formula I.

In a preferred embodiment of the present invention, the solvent is selected from the group consisting of acetone, methanol, water and any combinations thereof; the solvent is preferably acetone, or a mixed solvent consisting of methanol and water, or a mixed solvent consisting of acetone and water, wherein the volume ratio (ml/ml) of methanol and water is

- 4 045390 the mixed solvent consisting of methanol and water is 30:1-1:1, preferably 9:1, and the volume ratio (ml/ml) of acetone and water in the mixed solvent consisting of acetone and water is 6: 1-1:1, preferably 4:1.

In a preferred embodiment, the volume-to-weight ratio (ml/g) of solvent and compound of Formula I is 20:1-45:1 and the volume-to-mass ratio (ml/g) of water added to the solution of the compound of Formula I and the compound of Formula I is 20:1-90:1.

In a preferred embodiment of the present invention, the compound of formula I is dissolved in a solvent at 50-60°C.

In a preferred embodiment of the present invention, after dissolving the compound of formula I in the solvent, the resulting solution is stirred at room temperature for 0.5-24 hours.

In a preferred embodiment of the present invention, after adding water to a solution of a compound of formula I, the resulting solution is stirred at room temperature for 0.5-24 hours, then cooled to 5-15°C and stirring is continued for another 1-4 hours.

Method 2: Add a solvent to the compound of formula I, obtain a suspension of the compound of formula I by ultrasonication, protect the resulting suspension from light, stir, centrifuge and collect the solid phase to obtain crystalline Form 1 of the compound of formula I.

In a preferred embodiment, the solvent is selected from the group consisting of THF, methyl tert-butyl ether, water, acetone, isopropanol, dichloromethane, ethanol, and any combinations thereof.

In a preferred embodiment of the present invention, the suspension of the compound of formula I is stirred at room temperature or at 45-55°C, preferably at 50°C.

In a preferred embodiment of the present invention, the suspension of the compound of formula I is protected from light and stirred for 6-10 days.

Method 3: Add solvent to the compound of formula I, stir at 50-60°C until dissolved to obtain a solution of the compound of formula I, filter the resulting solution while still hot, then cool and crystallize the filtrate at -20 to 10°C, then centrifuge and collect the solids to obtain crystalline form 1 of the compound of formula I.

In a preferred embodiment, the solvent is selected from the group consisting of acetone, THF, dichloromethane, and any combinations thereof.

In a preferred embodiment of the present invention, after filtering the solution of the compound of formula I while it is still hot, the solution is slowly cooled to room temperature at a rate of 6°C/h, then cooled and crystallized at temperatures ranging from -20 to 10°C, preferably from 2 to 8°C.

Method 4: Add a first solvent to the compound of Formula I, then obtain a supersaturated solution of the compound of Formula I by ultrasonication, filter, then add a second solvent to the filtrate, stir, centrifuge and collect the solids to obtain crystalline Form 1 of the compound of Formula I.

In a preferred embodiment of the present invention, the first solvent is selected from the group consisting of methanol, ethanol, THF, acetone, isopropanol and any combinations thereof, the second solvent is selected from the group consisting of water, methyl tert-butyl ether, dichloromethane and any combinations thereof .

In a preferred embodiment, the volume ratio (ml/ml) of the first solvent and the second solvent is from 1:5 to 1:20, preferably 1:10.

Another object of the present invention is to provide crystalline salt forms of a compound of formula I, in particular a hydrochloride crystalline form, a sulfate crystalline form, a hydrobromide crystalline form, a phosphate crystalline form of a compound of formula I, which are hereinafter referred to as hydrochloride crystalline form A, sulfate crystalline form B, hydrobromide crystalline form C, phosphate crystalline form D of the compound of formula I.

The X-ray powder diffraction pattern of crystalline form A of the hydrochloride of the compound of formula I according to the present invention contains characteristic peaks at 2θ angles of 6.4±0.2°, 12.8±0.2°, 14.2±0.2° and 19 .0±0.2°.

In a preferred embodiment of the present invention, the X-ray powder diffraction pattern of crystalline Form A hydrochloride of the compound of formula I according to the present invention contains characteristic peaks at 2θ angles of 6.4±0.2°, 8.5±0.2°, 11.6± 0.2°, 12.8±0.2°, 14.2±0.2°, 17.1±0.2° and 19.0±0.2°.

In yet another preferred embodiment of the present invention, the X-ray powder diffraction pattern of crystalline Form A hydrochloride of the compound of formula I contains characteristic peaks at 2θ angles of 6.4±0.2°, 8.5±0.2°, 11.6±0 ,2°, 12.8±0.2°,

- 5 045390

14.2±0.2°, 17.l±0.2°, 19.0±0.2°, 19.7±0.2°, 21.3±0.2° and 24.5±0 ,2°.

Without being limited to anything, X-ray powder diffraction data of crystalline Form A hydrochloride of the compound of formula I according to the present invention are shown in table. 2.

table 2

Diffraction angle 20 Value d Relative intensity, % (Calculated according to peak height) 6.4 13.8 100 8.5 10.3 10.8 11.6 7.6 9 12.8 6.9 64.9 14.2 6.2 19.7 17.1 5.2 10.6 19.0 4.7 40.3 19.7 4.5 10.5 21.3 4.2 9.4 24.5 3.6 40.4

Without being limited to anything, the XRPD diffraction pattern of the hydrochloride crystal Form A of the compound of Formula I according to the present invention is shown in FIG. 5.

Without being limited to anything, a DSC thermogram of the hydrochloride crystal Form A of the compound of Formula I according to the present invention is shown in FIG. 6.

The present invention provides a method for preparing crystalline Form A hydrochloride of a compound of formula I. Specifically, said method is as follows: dissolving a compound of formula I in a solvent to obtain a solution of a compound of formula I, adding to said solution a compound of formula I with stirring hydrochloric acid in ethanol, stirring, centrifuging, collecting solids and drying to obtain hydrochloride crystalline Form A of the compound of formula I.

In a preferred embodiment, the compound of formula I is dissolved in a solvent by ultrasonication and heating.

In a preferred embodiment of the present invention, the solvent is selected from the group consisting of ethanol, acetone, acetonitrile, isopropanol, and any combinations thereof. In a preferred embodiment of the present invention, the concentration of hydrochloric acid in ethanol is 30-60 mg/ml, preferably 50 mg/ml.

In a preferred embodiment of the present invention, after adding hydrochloric acid in ethanol, the solution of the compound of formula I is stirred at room temperature for 4-24 hours.

The X-ray powder diffraction pattern of crystalline sulfate form B of the compound of formula I contains characteristic peaks at 2Θ angles of 12.2±0.2°, 17.1±0.2°, 18.4±0.2° and 20.1± 0.2°.

In a preferred embodiment of the present invention, the X-ray powder diffraction pattern of crystalline Form B sulfate of the compound of formula I contains characteristic peaks at 2Θ angles of 12.2±0.2°, 17.1±0.2°, 18.4±0.2 °, 19.6±0.2°, 20.1±0.2°, 20.6±0.2° and 22.1±0.2°.

In a preferred embodiment of the present invention, the X-ray powder diffraction pattern of crystalline Form B sulfate of the compound of formula I contains characteristic peaks at 2Θ angles of 12.2±0.2°, 17.1±0.2°, 18.4±0.2 °, 19.6±0.2°, 20.1±0.2°, 20.6±0.2°, 22.1±0.2°, 23.5±0.2°, 26.8 ±0.2° and 29.3±0.2°.

Without being limited to anything, X-ray powder diffraction data of crystalline Form B sulfate of the compound of formula I according to the present invention is shown in table. 3.

-6045390

Without being limited to anything, the XRPD diffraction pattern of the sulfate crystal Form B of the compound of Formula I according to the present invention is shown in FIG. 7.

Without being limited to anything, a DSC thermogram of the sulfate crystal Form B of the compound of Formula I according to the present invention is shown in FIG. 8.

Without being limited to anything, a TGA thermogram of crystalline Form B sulfate of the compound of Formula I according to the present invention is shown in FIG. 9.

Without being limited to anything, a graph of the DSP isotherm of the sulfate crystalline Form B of the compound of Formula I according to the present invention is shown in FIG. 10.

The present invention provides a method for preparing crystalline Form B sulfate of a compound of formula I. Specifically, said method is as follows: dissolving a compound of formula I in a solvent to obtain a solution of a compound of formula I, adding to said solution a compound of formula I with stirring sulfuric acid in ethanol, stirring, then centrifuging, collecting the solids and drying to obtain the sulfate crystalline Form B of the compound of formula I.

In a preferred embodiment of the present invention, the compound of formula I is dissolved in a solvent by ultrasonication and heating.

In a preferred embodiment of the present invention, the solvent is selected from the group consisting of ethanol, acetone, acetonitrile, isopropanol, and any combinations thereof.

In a preferred embodiment of the present invention, the concentration of sulfuric acid in ethanol is 30-60 mg/ml, preferably 50 mg/ml.

In a preferred embodiment of the present invention, after adding sulfuric acid in ethanol, the solution of the compound of formula I is stirred at room temperature for 4-24 hours.

The X-ray powder diffraction pattern of crystalline form C of hydrobromide of the compound of formula I according to the present invention contains characteristic peaks at 2θ angles of 6.3±0.2°, 12.6±0.2° and 18.9±0.2°.

In a preferred embodiment of the present invention, the X-ray powder diffraction pattern of crystalline form C of hydrobromide of the compound of formula I contains characteristic peaks at 2θ angles of 6.3±0.2°, 8.5±0.2°, 12.6±0.2 °, 18.9±0.2°, 21.3±0.2°, 24.4±0.2° and 25.2±0.2°.

Without being limited to anything, X-ray powder diffraction data of the hydrobromide crystal Form C of the compound of Formula I according to the present invention are shown in Table. 4.

- 7 045390

Table 4

Diffraction angle 20 Value d Relative intensity, % (Calculated according to peak height) 6.3 14.1 100 8.5 10.3 7.9 12.6 7.0 11.4 18.9 4.7 29.5 21.3 4.2 10.0 24.4 3.6 23.8 25.2 3.5 5.9

Without being limited to anything, the XRPD diffraction pattern of the hydrobromide crystal Form C of the compound of Formula I according to the present invention is shown in FIG. eleven.

Without being limited to anything, a DSC thermogram of the hydrobromide crystal Form C of the compound of Formula I according to the present invention is shown in FIG. 12.

The present invention provides a method for preparing the hydrobromide crystalline Form C of a compound of formula I. Specifically, said method is as follows: dissolving a compound of formula I in a solvent to obtain a solution of a compound of formula I, adding to said solution a compound of formula I with stirring hydrobromic acid in ethanol, stirring, then centrifuging, separating the solids and drying to obtain the hydrobromide crystalline Form C of the compound of formula I.

In a preferred embodiment of the present invention, the compound of formula I is dissolved in a solvent by ultrasonication and heating.

In a preferred embodiment of the present invention, the solvent is selected from the group consisting of ethanol, acetone, acetonitrile, isopropanol, and any combinations thereof.

In a preferred embodiment of the present invention, the concentration of hydrobromic acid in ethanol is 30-60 mg/ml, preferably 50 mg/ml.

In a preferred embodiment of the present invention, after adding hydrobromic acid in ethanol, a solution of the compound of formula I is stirred at room temperature for 4-24 hours.

The X-ray powder diffraction pattern of crystalline phosphate form D of the compound of formula I contains characteristic peaks at 2θ angles of 6.1 ± 0.2°, 10.9 ± 0.2° and 12.2 ± 0.2°.

In a preferred embodiment of the present invention, the X-ray powder diffraction pattern of crystalline Form D of phosphate of the compound of formula I contains characteristic peaks at 2θ angles of 6.1±0.2°, 10.9±0.2°, 11.7±0.2 ° and 12.2±0.2°.

Without being limited to anything, X-ray powder diffraction data of the phosphate crystal Form D of the compound of formula I according to the present invention are shown in Table. 5.

Table 5

Diffraction angle 20 Value d Relative intensity, % (Calculated according to peak height) 6.1 14.5 100 10.9 8.1 19.5 11.7 7.5 8.9 12.2 7.3 11.8

Without being limited to anything, the XRPD diffraction pattern of the crystalline phosphate form D of the compound of formula I according to the present invention is shown in FIG. 13.

Without being limited to anything, a DSC thermogram of the crystalline phosphate form D of the compound of formula I according to the present invention is shown in FIG. 14.

Without being limited to anything, a TGA thermogram of the crystalline phosphate form D of the compound of formula I according to the present invention is shown in FIG. 15.

- 8 045390

Without being limited to anything, a graph of the ADI isotherm of crystalline phosphate form D of the compound of formula I according to the present invention is shown in FIG. 16.

The present invention provides a method for preparing crystalline phosphate form D of a compound of formula I. Specifically, said method is as follows: a compound of formula I is dissolved in a first solvent to obtain a solution of a compound of formula I, and phosphoric acid in ethanol is added to said solution of a compound of formula I with stirring. , the solution is stirred, then centrifuged and the solid is collected, a second solvent is added to the collected solid, the solution is stirred, then centrifuged, the solid is collected and dried to obtain the phosphate crystalline Form D of the compound of formula I.

In a preferred embodiment of the present invention, a compound of formula I is dissolved in a first solvent by ultrasonication and heating.

In a preferred embodiment of the present invention, the first solvent is selected from the group consisting of ethanol, acetone, acetonitrile, isopropanol, and any combinations thereof; the second solvent is a mixed solvent consisting of acetone and water, where the volume ratio (ml/ml) of acetone and water is 7:1-9:1.

In a preferred embodiment of the present invention, the concentration of phosphoric acid in ethanol is 30-60 mg/ml, preferably 50 mg/ml.

In a preferred embodiment of the present invention, after adding phosphoric acid in ethanol, the solution of the compound of formula I is stirred at room temperature for 4-24 hours; after adding the second solvent, the resulting solution is stirred overnight at room temperature.

The present invention also provides a pharmaceutical composition containing crystalline Form 1 of a compound of Formula I, crystalline Form A hydrochloride of a compound of Formula I, crystalline Form B of a sulfate of a compound of Formula I, crystalline Form C of a hydrobromide of a compound of Formula I, and/or crystalline Form D of a phosphate of a compound of Formula I, as well as a pharmaceutical composition containing crystalline Form 1 of a compound of formula I, crystalline Form A of the hydrochloride of a compound of formula I, crystalline Form B of a sulfate of a compound of formula I, crystalline Form C of a hydrobromide of a compound of formula I, and/or crystalline Form D of a phosphate of a compound of formula I.

The present invention also provides the use of crystalline Form 1 of a compound of Formula I, crystalline Form A hydrochloride of a compound of Formula I, crystalline Form B of sulfate of a compound of Formula I, crystalline Form C of hydrobromide of a crystal of a compound of Formula I, and/or crystalline Form D of phosphate of a compound of Formula I for the preparation of medicaments. for the treatment of diseases or conditions associated with JAK1/TYK2, wherein the diseases or conditions are autoimmune diseases or disorders such as rheumatoid arthritis or inflammatory diseases or disorders, as well as cancers or tumor proliferative diseases or disorders.

Description of graphic materials

In fig. 1 shows an XRPD diffraction pattern of crystalline Form 1 of the compound of formula I according to the present invention.

In fig. 2A shows a DSC thermogram of crystalline Form 1 of the compound of formula I according to the present invention.

In fig. 2B shows another DSC thermogram of crystalline form 1 of the compound of formula I according to the present invention.

In fig. 3 shows a TGA thermogram of crystalline Form 1 of the compound of formula I according to the present invention.

In fig. 4 shows a graph of the ADI isotherm of crystalline Form 1 of the compound of formula I according to the present invention.

In fig. 5 shows an XRPD diffraction pattern of crystalline form A of the hydrochloride of the compound of formula I according to the present invention.

In fig. 6 shows a DSC thermogram of crystalline form A of the hydrochloride of the compound of formula I according to the present invention.

In fig. 7 shows an XRPD diffraction pattern of crystalline sulfate form B of the compound of formula I according to the present invention.

In fig. 8 shows a DSC thermogram of crystalline form B of sulfate of the compound of formula I according to the present invention.

In fig. 9 shows a TGA thermogram of crystalline Form B sulfate of the compound of formula I according to the present invention.

In fig. 10 is a graph of the DSP isotherm of crystalline sulfate form B of the compound of formula I according to the present invention.

In fig. 11 shows an XRPD diffraction pattern of the hydrobromide crystal Form C of the compound of formula I according to the present invention.

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In fig. 12 shows a DSC thermogram of crystalline form C of the hydrobromide of the compound of formula I according to the present invention.

In fig. 13 shows an XRPD diffraction pattern of crystalline phosphate form D of the compound of formula I according to the present invention.

In fig. 14 shows a DSC thermogram of crystalline phosphate form D of the compound of formula I according to the present invention.

In fig. 15 shows a TGA thermogram of crystalline phosphate form D of the compound of formula I according to the present invention.

In fig. 16 is a graph of the ADI isotherm of crystalline phosphate form D of the compound of formula I according to the present invention.

In fig. 17 shows an overlay of XRPD diffraction patterns of crystalline Form 1 of a compound of formula I according to the present invention after exposure to high temperature and accelerated aging conditions for 2 weeks.

In fig. 18 shows an overlay of XRPD diffraction patterns of crystalline phosphate form D of a compound of formula I according to the present invention after exposure to high temperature and accelerated aging conditions for 2 weeks.

In fig. 19 shows an overlay of DSC thermograms of crystalline Form 1 of the compound of formula I according to the present invention after aging at high temperature and under accelerated aging conditions for 2 weeks.

In fig. 20 shows an overlay of DSC thermograms of crystalline phosphate form D of a compound of formula I according to the present invention after exposure to high temperature and accelerated aging conditions for 2 weeks.

Embodiments of the Present Invention

The following examples further illustrate the invention, but the scope of the invention is not limited to these examples.

Information about the devices used when working on the present invention and the conditions for their use. __________________________________________________________________

No. Device Model Manufacturer Test procedure 1 High performance liquid chromatograph (HPLC) Agilent 1200, DAD Agilent Instrument: Agilent 1200 DAD HPLC System or similar configuration Column: Waters XBridge Shield RP18 4.6x150 mm, 3.5 µm Mobile phase: A: 0.05% aqueous solution of phosphoric acid; B: Acetonitrile 2 LC-MS Agilent 1200 Agilent Instrument: Agilent 1200 HPLC/6100 SQ System Column: Agilent XDB- C18, 4.6 mm χ 50 mm, 1.8 µm Mobile phase: A: 0.05% TFA in water; B: 0.05% TFA in acetonitrile 3 Ή NMR AVANCE III 400 MHz BRUKER Device: BRUKER AVANCE III 400 MHz UltraShield-Plus Digital NMR Spectroscopy Experiment: Experiment Ν PROTON 1H (default parameters) 4 X-ray D8 BRUKER Light source: CC.

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powder diffractometer (XRPD) Advance X-ray intensity: 40 kV/40 mA. Scan mode: Theta-theta. Scanning angle range: 4 40°. Scan step size: 0.05°. Scanning speed: 0.5 sec/step. 5 Differential scanning calorimeter (DSC) Q1000 TA Take a 2-4 mg sample, transfer it to an open aluminum container, equilibrate the sample in a nitrogen stream (50 ml/min) at 25°C and increase the temperature from 25°C to 300°C at a rate of 10°C/min. 6 Thermogravimetric analyzer (TGA) Q500 TA Take a 10-20 mg sample, transfer it to a platinum sample plate, from 25°C to 300°C at a rate of 10°C/min with a nitrogen flow over the sample of 60 ml/min and a balancing nitrogen flow of 40 ml/min. 7 Dynamic Vapor Sorption (DVA) Analyzer Advantag e 1 Surface Measurement System Take a sample weighed approximately 10 mg, set temperature by 25°C, dried for 60 minutes at a relative humidity of 0% and the indicators are determined moisture absorption of the sample when changing the relative humidity from 0% to 95% and indicators of moisture removal from the sample when changing relative humidity from 95% to 0%. Relative humidity change step: 5%. Mass balance is considered achieved when the rate of change of mass dm/dt becomes less than 0.002%. The criterion for balance in the test is the rate of mass change of less than 0.01%/min for 5 minutes, and the maximum time to achieve equilibrium is 2 hours.

- 11 045390

The materials and reagents used in the present invention are as follows.

Material Cleanliness/cleanliness class Batch number Manufacturer Compound of formula V >98.0% KM 1009- 1804001 ZINNOVA Compound of formula IV >98.0% KM 1008- 1804001 ZINNOVA N.N- diisopropylethylene (DIPED) ChDA KM261A- 1801002 Shanghai Qiao Chemical Science Co., Ltd. Ethanol / 160321047В Nanjing Chemical Reagent Co., Ltd. Methanol ChDA Р1176856 GENERAL-REAGENT Tetrahydrofuran Purity class: ChDA Р1167158 GENERAL-REAGENT Dichloromethane ChDA Р1216848 GENERAL-REAGENT Ethyl acetate(EA) ChDA Р1080359 GENERAL-REAGENT Acetone ChDA Р1160778 GENERAL-REAGENT Purified water Milli-Q Prepared in Milli-Q same day Methyl tert-butyl ether ChDA Р1135054 GENERAL-REAGENT Isopropanol vezhkh 6553IU13 Anhui Fulltime Specialized Solvent & Reagent Co., Ltd. Palladium on carbon (Pd/C) ChDA KM416A- 1603001 Shaanxi Rock New Materials Co., Ltd. (P)-lactamide ChDA KM 10121806001 Shanghai Zhongnai Biological Medical Technology Co., Ltd. Et ₃ O-BF ₄ ChDA KM416A- 1603001 Shaanxi Rock New Materials Co., Ltd. Hydrochloric acid ChDA GM007-1809001 Suzhou Zhushi Chemical Reagent Co., Ltd. Potassium carbonate Technically II GM009-1804001 Shanghai Weitang Industry Co., Ltd.

Examples

Preparation of a compound of formula III h ₂ n*^°

HCI

IV III

- 12 045390

Example 1. Preparation of the compound of formula III.

To a 25 mL three-neck flask was added ethanol (4 mL), compound of formula IV (0.20 g, 1.0 eq.), compound of formula V (0.18 g, 1.0 eq.) and DIPEA (0.39 g, 3.0 eq.), the system was stirred; the system was heated in a nitrogen atmosphere until vapor reflux (70-80°C) was achieved and stirred overnight under reflux at reflux temperature; then the system was cooled to room temperature (15-20°C), during cooling the solid phase precipitated; then water (4 ml) was added dropwise to the system, the system was stirred for 2 hours at room temperature (15-20°C), then filtered and the filter cake was washed with an aqueous solution of ethanol (2 ml, 1:1 v/v), the filter cake was dried under vacuum at a temperature of 45-50°C for 16 hours; About 0.21 g of solid was obtained with a LC-MS purity (at 214 nm) of 96.4% and a yield of 69%.

MS-ESI: [M+1]+: 303.1.

^1H NMR (400 MHz, CDCL3) δ 9.238 (s, 1H), 8.400 (d, 1H), 7.968 (d, 1H), 6.987 (d, 1H), 4.537-4.613 (m, 1H), 4.305-4.350 (m, 1H), 3.661-3.722 (m, 1H), 3.313-3.366 (m, 1H), 2.590-2.699 (m, 2H), 2.407-2.454 (m, 1H), 1.815-2.035 (m, 1H) , 1.688-1.806 (m, 2H).

Example 2 Preparation of a compound of formula III.

To a 250 mL three-neck flask was added ethanol (120 mL, 20 volumes), compound of formula IV (6.0 g, 1.0 eq.), compound of formula V (5.4 g, 1.01 eq.) and DIPEA ( 11.7 g, 3.0 eq.) and the system was stirred; then the system in a nitrogen atmosphere was heated to 70-80°C (internal temperature), the temperature was maintained while stirring the system for 8 hours; then the system was cooled to room temperature (15-20°C), during cooling the solid phase precipitated; then water (120 ml, 20 volumes) was added dropwise to the system, the system was stirred for 2 hours at room temperature (10-15°C), then filtered and the filter cake was washed with an aqueous solution of ethanol (30 ml, 1:1); the filter cake was dried under vacuum at 50°C for 16 hours; about 7.7 g of a yellow solid was obtained with an HPLC purity of 95.5% and a yield of 84.3%.

MS-ESI and 1H NMR data correspond to example 1.

Example 3 Preparation of a compound of formula III.

To a 25 mL three-neck flask was added ethanol (5 mL, 10 volumes), compound of formula IV (0.50 g, 1.0 eq.), compound of formula V (0.45 g, 1.01 eq.) and DIPEA ( 0.98 g, 3.0 eq.) and the resulting system was stirred; then the system was heated in a nitrogen atmosphere to 70-80°C, kept under reflux and stirred for 5 hours; then the system was cooled to room temperature (15-20°C), during cooling the solid phase precipitated; then water (5 ml, 10 volumes) was added dropwise to the system, the system was stirred for 2 hours at room temperature (10-15°C), then filtered and the filter cake was washed with an aqueous solution of ethanol (1:1) (1.5 ml, 3 volumes), the filter cake was dried under vacuum at 50°C for 16 hours; about 0.54 g of a brown solid was obtained with an HPLC purity of 95.4% and a yield of 71%.

MS-ESI and 1H NMR data correspond to example 1.

Example 4 Preparation of a compound of formula III.

To a 25 mL three-neck flask was added ethanol (5 mL, 10 volumes), compound of formula IV (0.50 g, 1.0 eq.), compound of formula V (0.45 g, 1.01 eq.) and DIPEA ( 0.72 g, 2.2 eq.) and the resulting system was stirred; then the system was heated in a nitrogen atmosphere to 70-80°C, kept under reflux and stirred for 5 hours; then the system was cooled to room temperature (15-20°C), during cooling the solid phase precipitated; then water (7.5 ml, 15 volumes) was added dropwise to the system, the system was stirred for 1 hour at room temperature (10-15°C), then cooled to 5-10°C and stirred for 2 hours, after After this, the system was filtered and the filter cake was washed with an aqueous solution of ethanol (1:1) (1.5 ml, 3 volumes), the filter cake was dried under vacuum at 50°C for 16 hours; about 0.57 g of a brown solid was obtained with an HPLC purity of 91.4% and a yield of 75%.

MS-ESI and 1H NMR data correspond to example 1.

Example 5 Preparation of a compound of formula III.

To a 250 mL three-neck flask was added ethanol (50 mL, 10 volumes), compound of formula IV (5.0 g, 1.0 eq.), compound of formula V (4.5 g, 1.01 eq.) and DIPEA ( 7.2 g, 2.2 eq.) and the resulting system was stirred; then the system was heated in a nitrogen atmosphere to 70-80°C, kept under reflux and stirred for 5 hours; then the system was cooled to room temperature (15-20°C), during cooling the solid phase precipitated; then water (75 ml, 15 volumes) was added dropwise to the system, the system was stirred for 1 hour at room temperature (10-15°C), then cooled to 5-10°C and stirred for 2 hours, after which the system filtered and the filter cake was washed with an aqueous solution of ethanol (1:1, 15 ml), the filter cake was dried under vacuum at 50°C for 16 hours; about 6.6 g of a yellow solid was obtained with an HPLC purity of 94.2% and a yield of 86.7%.

MS-ESI and 1H NMR data correspond to example 1.

Example 6 Preparation of a compound of formula III.

To a 500 mL three-neck flask was added ethanol (180 mL, 10 volumes), compound of formula IV (17.8 g, 1.0 eq.), compound of formula V (16.0 g, 1.01 eq. ) and DIPEA (25.7 g, 2.2 eq.) and the resulting system was stirred; then the system was heated in a nitrogen atmosphere to 70-80°C, kept under reflux and stirred for 5 hours; then the system was cooled to room temperature (15-20°C), during cooling the solid phase precipitated; then water (270 ml, 15 volumes) was added dropwise to the system, the system was stirred for 1 hour at room temperature (10-15°C), then cooled to 5-10°C and stirred for 2 hours, after which the system filtered and the filter cake was washed with an aqueous solution of ethanol (ethanol/water=1:1.5, v/v, 40 ml), the filter cake was dried under vacuum at 50°C for 16 hours; about 23.0 g of a brown solid was obtained with an HPLC purity of 95.3% and a yield of 85.2%.

MS-ESI and 1H NMR data correspond to example 1.

Example 7 Preparation of a compound of formula III.

To a 3000 mL three-neck flask was added ethanol (1000 mL, 10 volumes), compound of formula IV (100 g, 1.0 eq.), compound of formula V (89.9 g, 1.01 eq.) and DIPEA (143, 2 g, 2.2 eq.), and the resulting system was stirred; the system was then heated under a nitrogen atmosphere to 85-90°C (internal temperature approximately 75°C), refluxed and stirred for 10 hours; then the system was cooled to room temperature (15-20°C), during cooling the solid phase precipitated; then water (1500 ml, 15 volumes) was added dropwise to the system, the system was stirred for 1 hour at room temperature (10-15°C), then cooled to 5-10°C and stirred for 2 hours, after which the system filtered and the filter cake was washed with an aqueous solution of ethanol (1:1.5, v/v, 200 ml), the filter cake was dried under vacuum at 50°C for 16 hours; about 130 g of a reddish-brown solid were obtained with an HPLC purity of 94.2% and a yield of 85.5%.

MS-ESI and 1H NMR data correspond to example 1.

Example 8 Preparation of the compound of formula III.

To a 5000 mL three-neck flask was added ethanol (2000 mL, 10 volumes), compound of formula IV (200 g, 1.0 eq), compound of formula V (179.7 g, 1.01 eq), and DIPEA (286, 4 g, 2.2 eq.) and the resulting system was stirred; the system was then heated under a nitrogen atmosphere to 70-80°C (internal temperature approximately 65-70°C), refluxed and stirred for 16 hours; then the system was cooled to room temperature (15-20°C), during cooling the solid phase precipitated; then water (3000 ml, 15 volumes) was added dropwise to the system, the system was stirred for 1 hour at room temperature (10-15°C), then cooled to 5-10°C and stirred for 2 hours, after which the system filtered and the filter cake was washed with an aqueous solution of ethanol (1:1.5, v/v, 400 ml), the filter cake was dried under vacuum at 50°C for 16 hours; about 251 g of a reddish-brown solid were obtained with an HPLC purity of 93.4% and a yield of 78.1%.

MS-ESI and 1H NMR data correspond to example 1.

Example 9. Preparation of the compound of formula III.

To a 20,000 mL three-neck flask was added ethanol (5,000 mL, 10 volumes), compound of formula IV (500 g, 1.0 eq), compound of formula V (450 g, 1.01 eq), and DIPEA (723 g, 2 ,2 eq.) and the resulting system was stirred; the system was then heated under a nitrogen atmosphere to 80-90°C (internal temperature approximately 70-80°C), refluxed and stirred for 16 hours; then the system was cooled to room temperature (25-30°C), during cooling the solid phase precipitated; then water (7500 ml, 15 volumes) was added dropwise to the system, the system was stirred for 1 hour at room temperature (25-30°C), then cooled to 10-15°C and stirred for 2 hours, after which the system filtered and the filter cake was washed with an aqueous solution of ethanol (1:1.5, v/v, 1000 ml), the filter cake was dried under vacuum at 50-55°C for 24 hours; about 623 g of product were obtained with an HPLC purity of 93.7%, residual ethanol of about 0.5%, product content of 93.1% and yield of 76.2%.

MS-ESI and 1H NMR data correspond to example 1.

Example 10 Preparation of the compound of formula III.

To a 500 mL three-neck flask was added ethanol (100 mL, 10 volumes), compound of formula IV (10.0 g, 1.0 eq.), compound of formula V (9.0 g, 1.01 eq.) and DIPEA ( 14.3 g, 2.2 eq.) and the resulting system was stirred; then the system was heated in a nitrogen atmosphere to 70-80°C, kept under reflux and stirred for 16 hours; then the system was cooled to room temperature (20-30°C), during cooling the solid phase precipitated; then water (150 ml, 15 volumes) was added dropwise to the system, the system was stirred for 2 hours at room temperature (20-30°C), then cooled to 5-10°C and stirred for 2 hours, after which the system filtered and the filter cake was washed with an aqueous solution of ethanol (1:1.5, v/v, 25 ml), the filter cake was dried in an oven under vacuum at a temperature of 50-55°C for 16 hours; about 13.7 g of product were obtained with an HPLC purity of 93.7% and a yield of 90%.

MS-ESI and 1H NMR data correspond to example 1.

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Example 11 Preparation of the compound of formula III.

To reactor R0462 was added ethanol (17 kg, 10 volumes), compound of formula IV (2.2 kg, 1.0 eq), compound of formula V (1.98 kg, 1.01 eq) and DIPEA (3.19 kg, 2.2 eq.) and the resulting system was stirred; then the system was heated in a nitrogen atmosphere to 75-80°C (internal temperature about 70-80°C), stirred for 16 hours, after which the system was cooled to room temperature (15-25°C), upon cooling a solid precipitate formed; then water (33 kg, 15 volumes) was added dropwise to the system, the system was stirred for 2 hours at room temperature (10-15°C), then cooled to 5-10°C and stirred for 4 hours; After this, the system was filtered, the filter cake was washed with an aqueous solution of ethanol (ethanol/water = 1:2, v/v, 6.2 kg), then the filter cake was dried under vacuum <0.08 MPa at a temperature in a water jacket about 45-55°C for 16 hours; 2.64 kg of a brown solid was obtained with an HPLC purity of 94.0%, a major content of 93.4% and a yield of 79.04%.

MS-ESI and ¹ H NMR data correspond to example 1.

Preparation of the compound of formula II.

Ill II

Example 12 Preparation of a compound of formula II.

Compound of formula III (5.0 g), THF (50 mL, 10 volumes) and palladium on carbon (0.75 g, 10% Pd/C with 50% humidity) were added sequentially to a 100 mL stainless steel high-pressure reactor. ; the system was purged 5 times with nitrogen and then 5 times with hydrogen; the pressure in the system was increased to 0.50 MPa with hydrogen, the system was heated to 25-35°C and the temperature was maintained with constant stirring of the system for 24 hours; the reaction solution was filtered with diatomaceous earth, the filter cake was washed with THF (20 ml), the filtrate was concentrated to dryness to give 4.2 g of a brown solid with an HPLC purity of 94.9% and a yield of 93.3%.

MS-ESI: [M+1]+: 273.1.

^1H NMR (400 MHz, CDCl3) δ 7.988 (s, 1H), 7.688 (d, 1H), 6.805 (d, 1H), 4.190-4.338 (m, 3H), 3.584-3.648 (m, 1H), 3.147 -3.206 (t, 1H), 2.594-2.651 (d, 2H), 2.318-2.364 (m, 1H), 1.917-1.974 (m, 1H), 1.633-1.738 (m, 1H), 1.456-1.525 (m, 1H).

Example 13 Preparation of a compound of formula II.

To a 5000 ml high pressure stainless steel reactor, compound of formula III (120.0 g), THF (2400 ml, 20 volumes) and palladium on carbon (18 g, 10% Pd/C with 50% humidity) were added sequentially; the system was purged 5 times with nitrogen and then 5 times with hydrogen; the pressure in the system was increased to 0.50 MPa with hydrogen, the system was heated to 25-35°C and the temperature was maintained with constant stirring of the system for 24 hours; the reaction solution was filtered with diatomaceous earth, the filter cake was washed with THF (600 ml) (until the fluorescent signal on the TLC plates almost completely disappeared), the filtrate was concentrated, resulting in 130 g of a black oily solid with an HPLC purity of 91.7% and yield 120.26%.

MS-ESI and ¹ H NMR data correspond to Example 12.

Example 14 Preparation of a compound of formula II.

Compound of formula III (100.0 g), THF (2000 ml, 20 volumes) and palladium on carbon (15.0 g, 10% Pd/C with 50% humidity) were added sequentially to a 5 L high pressure stainless steel reactor. ; the system was purged 5 times with nitrogen and then 5 times with hydrogen; the pressure in the system was increased to 0.5-1.0 MPa with hydrogen, the temperature of the water jacket was set to 30°C and the system was stirred, maintaining the specified temperature for 16 hours, then the reaction solution was filtered using diatomaceous earth, the filter cake was washed with THF (1000 ml ), resulting in 3877 g of a solution of the compound of formula II in THF.

Post-treatment 1: The above filtrate (1820 g, approximately 40 g of compound of formula II, calculated on the basis of 100% yield) was concentrated to 2-3 volumes (80-120 ml) on a rotary evaporator, then the system was changed from solvent to ethanol ( 150 mlx2) to 2-3 volumes (80-120 ml), resulting in 78 g of a solution of the compound of formula II in ethanol with a content of 47.25% and a yield of 92.14%.

Post-workup 2: The above filtrate (450 g containing approximately 10 g of compound of formula II based on 100% yield) was concentrated to dryness on a rotary evaporator to obtain 10.5 g of a brownish-red solid.

Post-treatment 3: The above filtrate (450 g, approximately 10 g of compound of formula II based on 100% yield) was placed in a flask and concentrated to approximately 30-40 ml (3-4 volumes)

- 15 045390 on a rotary evaporator; the remainder of the concentrate was subjected to solvent replacement with ethanol (50 mlx2), bringing the volume to approximately 30-40 ml (3-4 volumes), resulting in a black oily concentrate residue, which was directly sent to the next stage of the reaction.

MS-ESI and ¹ H NMR data correspond to example 12.

Example 15 Preparation of a compound of formula II.

To a 500 mL three-neck flask, THF (240 mL, 20 volumes), compound of formula III (12.0 g), and palladium on carbon (1.8 g, 5% Pd/C with 50% moisture water) were added sequentially; the system was purged 5 times with nitrogen and then 5 times with hydrogen, stirred for 48 hours at room temperature (25-30°C) and a hydrogen pressure of about 0.1 MPa; the reaction solution was filtered, the filter cake was washed with THF (60 ml), the combined filtrate was concentrated to 20-30 ml on a rotary evaporator, then the solvent was replaced with ethanol (60 mlx2), bringing the volume to 20-30 ml, resulting in 24 g of the compound of formula II in ethanol, the solution was used directly in the next reaction step.

MS-ESI and %NMR data are consistent with Example 12.

Example 16 Preparation of a compound of formula II.

To a 5000 ml three-neck flask, THF (1500 ml, 15 volumes), compound of formula III (100 g) and palladium on carbon (15 g, 5% Pd/C with 50% humidity water) were added sequentially; the system was purged 5 times with nitrogen and then 5 times with hydrogen, stirred for 48 hours at room temperature (20-25°C) and a hydrogen pressure of about 0.1 MPa; the reaction solution was filtered and the filter cake washed with THF (200 ml), the combined filtrate was concentrated to 200-300 ml on a rotary evaporator, resulting in 185.6 g of the compound of formula II in THF with an HPLC purity of 94.2%, content 43.2% and yield 94.0%.

MS-ESI and %NMR data are consistent with Example 12.

Example 17 Preparation of a compound of formula II.

To a 20,000 mL three-neck flask, THF (12,400 mL, 20 volumes), compound of formula III (620 g), and palladium on carbon (93 g, 5% Pd/C with 50% moisture water) were added sequentially; the system was purged 5 times with nitrogen and then 5 times with hydrogen, stirred for 48 hours at room temperature (30-35°C) and a hydrogen pressure of about 0.1 MPa; the reaction solution was filtered using diatomaceous earth (200 g), the filter cake was washed with THF (1200 ml), the combined filtrate was concentrated to 1200-1800 ml on a rotary evaporator, resulting in 1664 g of the compound of formula II in THF with an HPLC purity of 93.8%, content 34.57% and yield content 110.6%.

MS-ESI and %NMR data are consistent with Example 12.

Example 18 Preparation of a compound of formula II.

To a 250 ml high pressure reactor was added THF (140 ml, 70 volumes), compound of formula III (2.0 g) and palladium on carbon (0.3 g, 5% Pd/C with 50% humidity); the high-pressure reactor was closed and the nut was tightened tightly; the system was purged 3 times with nitrogen and then 3 times with hydrogen; then hydrogen was pumped into the high-pressure reactor to a pressure of approximately 0.50 ± 0.05 MPa, the inlet valve was closed; the mixer was started at a rotation speed of 500 rpm; the hydrogen pressure in the high-pressure reactor was maintained at 0.5±0.05 MPa at a temperature of 25-35°C, stirring in the high-pressure reactor and the reaction in the system were maintained for 96 hours; then the reaction solution was filtered using diatomite (10 g), the filter cake was washed with THF (60 ml); the combined filtrate was concentrated to dryness by rotary evaporation to obtain 1.8 g of a semi-solid oily substance with an HPLC purity of 91.2% and a yield of 99.9%.

MS-ESI and %NMR data are consistent with Example 12.

Example 19 Preparation of a compound of formula II.

THF (167 kg, 70 volumes), compound of formula III (2.64 kg) and palladium on carbon (0.4 kg, 5% Pd/C with 50% humidity) were added to a 500 L high pressure reactor; the system was purged 5 times with nitrogen and then 5 times with hydrogen; then hydrogen was pumped into the high-pressure reactor to a pressure of approximately 0.50 ± 0.05 MPa, the inlet valve was closed; started the mixer; the hydrogen pressure in the high-pressure reactor was maintained at 0.5±0.05 MPa at a temperature of 25-35°C, stirring in the high-pressure reactor and the reaction in the system were maintained for 120 hours; the reaction solution was pressed on a filter, the filter cake was washed with THF (13 kg); the combined filtrate was distilled under reduced pressure (to 2-3 volumes), resulting in 11 kg of compound of formula II in THF with an HPLC purity of 90.7%, a content of 18.5%, and a yield of 91.9%.

MS-ESI and ¹ H NMR data correspond to example 12.

Example 20 Preparation of a compound of formula II.

To a 100 mL high pressure stainless steel reactor was added THF (60 mL, 12 volumes), compound of Formula III (5.0 g), and palladium on carbon (0.75 g, 5% Pd/C with 50% humidity); the system was purged 5 times with nitrogen and then 5 times with hydrogen; then hydrogen was pumped into the high-pressure reactor

- 16 045390 to a pressure of approximately 0.5-1.0 MPa, the jacket temperature was set to 30°C, and the system was stirred, maintaining this temperature, for 42 hours; after completion of the reaction, the reaction solution was filtered using diatomite, the filter cake was washed with THF (100 ml); this gave 197.8 g of the compound of formula II in THF; the solution was concentrated on a rotary evaporator to 2-3 volumes (10-15 ml); then the solvent in the system was replaced with ethanol (25 mlx2), bringing to 2-3 volumes (10-15 ml); the resulting compound of formula II in ethanol was directly used in the next reaction step.

MS-ESI and Ή NMR data are consistent with Example 12.

Preparation of a compound of formula I.

II I

Example 21 Preparation of a compound of formula I.

THF (60 ml, 12 volumes), (R)-lactamide (6.6 g, 4.0 eq.) and Et ₃ O-BF ₄ (13.9 g, 4.0 eq.), the system was stirred; then the materials in three-neck flask No. 1 were mixed under a nitrogen atmosphere and left for later use; to another three-neck flask (No. 2) with a volume of 250 ml, the compound of formula II (5.0 g, 1.0 eq.) and ethanol (80 ml, 16 volumes) were added; the system was heated to 70±5°C in a nitrogen atmosphere; then the materials from three-neck flask No. 1 were added dropwise to three-neck flask No. 2 using a syringe for 10-20 minutes; to carry out the reaction, the system was heated to 85±5°C (internal temperature was in the range of 72-75°C) in a nitrogen atmosphere for 2 hours; then the system was cooled to room temperature; the reaction liquid was concentrated on a rotary evaporator until the distillation of the liquid fraction was almost complete; a solution of 1 M HCl (80 ml) was added to the residual concentrated liquid, and the pH of the solution was approximately 1 (determined using indicator paper); the system was then extracted four times with DCM (50 mlx4); The pH of the aqueous phase was adjusted to 7-8 with a saturated sodium bicarbonate solution; the system was stirred at room temperature for 0.5 h, then filtered, the filter cake was washed with water (60 ml) and EA (10 ml), respectively; then the filter cake was dried in vacuum at 50°C for 16 hours; the result was 4.3 g of a pale yellow solid with a purity of 95.0%; said solid was dissolved in methanol (30 ml); 4.1 g of a silicon-based magnetic separator for metallic inclusions and 1.0 g of activated carbon were added, the system was heated to 50°C and stirred for 1 hour, then cooled, filtered, and washed with methanol (30 ml); the resulting filtrate was concentrated on a rotary evaporator until the distillation of the liquid fraction was almost complete, and methanol (10 ml) and MTBE (25 ml) were added to the residue; the system was heated to 50°C and stirred for 0.5 h and left to cool, then the system was cooled to 10 ± 5°C and stirred for 0.5 h, after which it was filtered; the filter cake was washed with MTBE (25 ml); the filter cake was then dried under vacuum at 50°C for 16 hours to obtain 3.2 g of a pale yellow solid with a purity of 97.9%.

MS-ESI: [M+1]+: 327.6.

^1H NMR (400 MHz, CDCl ₃ ) δ 8.988 (s, 1H), 7.922 (d, 1H), 7.175 (d, 1H), 5.200-5.265 (m, 1H), 4.859-4.942 (m, 1H), 4.350-4.406 (t, 1H), 4.020-4.108 (m, 2H), 3.067 (d, 1H), 2.619-2.779 (m, 3H), 2.108-2.269 (m, 2H), 1.790-1.895 (m, 3H ).

Example 22 Preparation of a compound of formula I.

THF (650 ml, 12 volumes), (R)-lactamide (70.6 g, 4.0 eq.) and Et ₃ O-BF ₄ (150.6 g, 4.0 eq.), the system was stirred; then the materials in three-neck flask No. 1 were mixed under a nitrogen atmosphere and left for later use; to another 2000 ml three-neck flask (No. 2) was added the compound of formula II (54 g, 1.0 eq.) and ethanol (860 ml, 16 volumes); the system was heated to 70±5°C in a nitrogen atmosphere; then the materials from three-neck flask No. 1 were slowly added dropwise to three-neck flask No. 2 using a syringe over 1 hour; to carry out the reaction, the system was heated to 85±5°C (internal temperature was in the range of 72-75°C) in a nitrogen atmosphere for 2 hours; then the system was cooled to room temperature; the reaction liquid was concentrated on a rotary evaporator until the distillation of the liquid fraction was almost complete; a solution of 1 M HCl (450 ml) was added to the residual concentrated liquid, and the pH of the solution was approximately 1 (determined using indicator paper); the system was then extracted four times with DCM (270 mlx4); The pH of the aqueous phase was adjusted to 7-8 with a saturated sodium bicarbonate solution; the system was stirred at room temperature for 0.5 h, then filtered, the filter cake was washed with water (540 ml), MTBE (270 ml) was added to the filter cake, sys.

- 17 045390 the mixture was stirred at room temperature for 0.5 h, then filtered, the filter cake was washed with MTBE (108 ml); then the filter cake was dried in vacuum at 50°C for 16 hours; the result was 49.2 g of a light yellow solid with an HPLC purity of 94.2%; said solid was dissolved in methanol (380 ml); a magnetic separator of metallic inclusions based on silicon (44 g) and activated carbon (5.4 g) were added, the system was heated to 50°C and stirred for 1 hour, then cooled, filtered, and washed with methanol (430 ml); the resulting filtrate was concentrated on a rotary evaporator to a volume of approximately 80-110 ml (1.5-2 volumes) and MTBE (540 ml) was added to the residue; the system was heated to 50°C and stirred for 1 hour, then cooled to 10±5°C and stirred for 0.5 hour, after which it was filtered; the filter cake was washed with MTBE (270 ml); resulting in 42.4 g of a filter cake with an HPLC purity of 96.9%, the filter cake was then dried under vacuum at 50° C. for 16 hours, resulting in 41.0 g of a light yellow solid with purity according to HPLC equal to 96.7% and yield 63.3%.

Purification of a compound of formula I.

The compound of formula I (41 g) was dissolved in methanol; Silica gel (50 g) was added to the solution, the system was concentrated to dryness and left for subsequent use; Silica gel (200 g) was loaded into the chromatography column, and the column was compacted using an air pump; then a compound of formula I mixed with silica gel was added to the chromatography column, the column was compacted using an air pump; after that, the chromatographic column was eluted with an eluent (V _MeOH : VD _CM = 1:100-1:30); qualified components were selected, concentrated to dryness and the product was dried under vacuum at 50°C for 16 hours; this gave 36 g of an almost white solid with an HPLC purity of 98.5%.

MS-ESI and 1H NMR data correspond to example 21.

Example 23 Preparation of a compound of formula I.

THF (60 ml, 6 volumes), (R)-lactamide (13.2 g, 4.0 eq.) and Et ₃ O-BF ₄ (27.9 g, 4.0 eq.), the system was stirred; then the materials in flask No. 1 were mixed under a nitrogen atmosphere and left for later use; to another 250 ml three-neck flask (No. 2) was added the compound of formula II (10 g, 1.0 eq.) and ethanol (100 ml, 10 volumes); the system was heated to 70±5°C in a nitrogen atmosphere; then the materials from three-neck flask No. 1 were slowly added dropwise to three-neck flask No. 2 over 20 minutes; to carry out the reaction, the system was heated to 80±5°C (internal temperature was in the range of 72-75°C) in a nitrogen atmosphere for 0.5 hours; then the system was cooled to room temperature (20-30°C); the reaction liquid was concentrated to a volume of approximately 50-80 ml on a rotary evaporator at a temperature in the range of 30-40°C; then water was added to the system (100 ml, 10 volumes), then the system was concentrated using a rotary evaporator at a temperature in the range of 30-40°C until the distillation of the liquid fraction was almost complete; the system was cooled to 20-30°C, after which the system temperature was controlled within 20-30°C, a solution of 12 M HCl (5.5 g) was used to bring the pH of the system to 2-3, then the system was extracted with ethyl acetate (50 mlx2, 5 volumesx2); the organic phase was separated and the aqueous phase was transferred to a flask; the system temperature was controlled within 20-30°C, and the pH was adjusted to 8-9 with a saturated solution of potassium carbonate (23 g); then the temperature of the system was controlled within 20-25°C, the system was stirred for 2 hours, then filtered, the filter cake was washed with water (50 ml) and MTBE (50 ml); the filter cake was then blow-dried at 50° C. for 24 hours to obtain 18 g of an earthy yellow solid with an HPLC purity of 93.5%.

MS-ESI and 1H NMR data correspond to example 21.

Example 24 Preparation of a compound of formula I.

THF (120 ml, 12 volumes), (R)-lactamide (13.2 g, 4.0 eq.) and Et ₃ O-BF ₄ (27.8 g, 4.0 eq.), the system was stirred; then the materials in flask No. 1 were mixed under a nitrogen atmosphere and left for later use; to another 500 ml three-neck flask (No. 2) was added the compound of formula II (10 g, 1.0 eq.) and ethanol (140 ml, 14 volumes); the system was heated to 40-45°C (internal temperature) in a nitrogen atmosphere; then the materials from three-neck flask No. 1 were added dropwise to three-neck flask No. 2 for 1 hour; to carry out the reaction, the system was maintained at a temperature of 40-45°C (internal temperature) in a nitrogen atmosphere for 4.5 hours; then the system was cooled to room temperature and water (20 ml, 2 volumes) was added; the reaction liquid was concentrated on a rotary evaporator at a temperature in the range of 30-40°C until the distillation of the liquid fraction was almost complete; then the system was cooled to 20-30°C, after which the system temperature was controlled within 20-30°C, a solution of 12M HCl (3 ml) was used to bring the pH of the system to 2-3, then the system was extracted with ethyl acetate (50 mlx2, 5 volumesx2 ); the organic phase was separated and the aqueous phase was transferred to a flask; the system temperature was controlled within 20-30°C, and the pH was adjusted to 8-9 using a 50% potassium carbonate solution (15 ml); then the temperature of the system was controlled within 20-25°C, the system was stirred for 2 hours, then filtered, the filter cake was washed with water (50 ml) and acetone (50 ml); the crude product was triturated and mixed with water (50 ml) at 20-25°C for 1 hour, then the system was filtered and the filter cake was washed with water (50 ml) and acetone (50 ml), the filter cake was dried in a blower at 50° C for 24 hours, resulting in 17.8 g of a khaki solid with an HPLC purity of 95.3%.

MS-ESI and 1H NMR data correspond to example 21.

Example 25 Preparation of a compound of formula I.

THF (60 ml, 12 volumes), (R)-lactamide (6.6 g, 4.0 eq.) and Et ₃ O-BF ₄ (13.9 g, 4.0 eq.), the system was stirred; then the materials in flask No. 1 were mixed under a nitrogen atmosphere and left for later use; to another 250 ml three-neck flask (No. 2) was added the compound of formula II (5 g, 1.0 eq.) and ethanol (70 ml, 14 volumes); the system was heated to 40-45°C (internal temperature) in a nitrogen atmosphere; then the materials from three-neck flask No. 1 were added dropwise to three-neck flask No. 2 for 20 minutes; to carry out the reaction, the system was maintained at a temperature of 40-45°C (internal temperature) in a nitrogen atmosphere for 3 hours; then the system was cooled to room temperature and filtered, the filter cake was washed with THF (10 ml), and water (10 ml, 2 volumes) was added to the filtrate; then the filtrate was concentrated on a rotary evaporator to a volume of 10-20 ml (2-4 volumes), the concentrated residue was subjected to replacement of the solvent with ethyl acetate (25 mlx2) and concentrated to a volume of 10-20 ml (2-4 volumes), water was added to the concentrated residue (50 ml, 10 parts by volume), the internal temperature of the system was controlled within 20-25°C, a solution of 12 M HCl (4.1 g) was used to adjust the pH of the system to 1-2, then activated carbon (0. 5 g) and the system was stirred for 2 hours, then filtered, the filter cake was washed with water (10 ml) and a solution of 1 M HCl (10 ml); the combined filtrate was extracted with ethyl acetate (25 mlx2), the organic phase was removed, then the internal temperature of the system was controlled between 20-25°C and the pH was adjusted to 9-10 with saturated potassium carbonate solution (15 g); then the internal temperature of the system was controlled within 15-20°C, the system was stirred for 1 hour and filtered, the filter cake was washed with water (10 ml), then it was triturated in an aqueous-acetone solution (50 ml, V/V=1:1 ) for 1 hour, the system was filtered, and the filter cake was washed with aqueous acetone solution (10 ml, V/V=1:1); The filter cake was blow dried at 50° C. for 24 hours to obtain 5.0 g of a pale gray solid with an HPLC purity of 95.6% and a yield of 83.5%.

Purification of a compound of formula I.

5.0 g of the resulting solid and methanol (40 ml) were added to the flask, the mixture was stirred for 10 min at room temperature, the materials were mostly dissolved and a clear solution was obtained; then activated carbon (0.5 g) and silica gel (4.0 g) were added to the system; the system was heated to 50-55°C and the temperature was maintained while stirring the system for 2 hours, then the system was filtered using silica gel (5 g), the filter cake was washed with methanol (50 ml); the filtrate was concentrated on a rotary evaporator to a volume of 5-10 ml; MTBE (50 ml) was added to the concentrated residue; the system was heated at reflux and kept at reflux for 1 hour; then the system was cooled to 5-10°C, the temperature was maintained at this level and the system was stirred for 1 hour, after which it was filtered, the filter cake was washed with MTBE; the filter cake was dried in an oven under vacuum at 50°C for 16 hours; the result was 3.0 g of an almost white solid with a yield of 60% and a purity of 97.9%; the filtrate was concentrated to dryness to give 1.4 g of a yellow solid.

MS-ESI and 1H NMR data correspond to example 21.

Example 26 Preparation of a compound of formula I.

THF (55 ml, 12 volumes), (R)-lactamide (5.9 g, 4.0 eq.) and Et ₃ O-BF ₄ (12.6 g, 4.0 eq.), the system was stirred; then the materials in flask No. 1 were mixed under a nitrogen atmosphere and left for later use; to another 250 ml three-neck flask (No. 2) was added the compound of formula II (4.5 g, corresponding to 5.0 g of the compound of formula III in example 20 based on 100% yield) and ethanol (70 ml, 15.5 volumes); the system was heated under reflux until reflux began in a nitrogen atmosphere; then the materials from three-neck flask No. 1 were added dropwise to three-neck flask No. 2 in one session; to carry out the reaction, the system was heated to a temperature of 85±5°C (internal temperature 74-76°C) in a nitrogen atmosphere for 1 hour; then the system was cooled to room temperature and filtered, the filter cake was washed with THF (10 ml), and water (10 ml, 2 volumes) was added to the filtrate; then the filtrate was concentrated on a rotary evaporator to a volume of 10-20 ml (2-4 volumes), the concentrated residue was subjected to replacement of the solvent with ethyl acetate (25 mlx2) and concentrated to a volume of 10-20 ml (2-4 volumes), water was added to the concentrated residue (50 ml, 10 parts by volume), the internal temperature of the system was controlled within 20-25°C, a solution of 12 M HCl (3.6 g) was used to adjust the pH of the system to 1-2, then activated carbon (0. 5 g) and the system was stirred at room temperature for 2 hours, then filtered, the filter cake was washed with water (10 ml) and a solution of 1 M HCl

- 19 045390 (10 ml); the combined filtrate was extracted with ethyl acetate (25 mlx2), the organic phase was separated, then the internal temperature of the system was controlled between 20-25°C and the pH was adjusted to 9-10 with a saturated solution of potassium carbonate (15 g); then the internal temperature of the system was controlled within 15-20°C, the system was stirred for 1 hour and filtered, the filter cake was washed with water (10 ml), then it was triturated in an aqueous-acetone solution (50 ml, V/V=1:1 ) for 1 hour, the system was filtered, and the filter cake was washed with aqueous acetone solution (10 ml, V/V=1:1); The filter cake was blow dried at 50° C. for 16 hours to obtain 4.9 g of a pale gray solid with an HPLC purity of 94.5%.

Purification of a compound of formula I.

The resulting solid (4.9 g) was placed in a flask and ethanol (100 ml) was added, the mixture was stirred at room temperature for 10 minutes, the materials were mostly dissolved, and a clear solution was obtained; then silica gel (5.0 g, 1X) was added to the system, the system was concentrated to dryness on a rotary evaporator, and left for subsequent use; the crude compound of formula I mixed with silica gel was passed through a silica gel column (40 g, 8X), the column was eluted with a mixed solution of ethyl acetate and ethanol (V _EA :V _EtOH =100:1-30:1); the components were checked by TLC, and the components containing the product were collected and concentrated to dryness; the result was 0.5 g of a white product with a purity of 95.0% and 2.9 g of a pale yellow product with a purity of 98.7%; then ethyl acetate (30 ml) and ethanol (3 ml) were added to the flask containing 2.9 g of product; the system was refluxed to boiling and left to boil for 1 hour; then the system was cooled to 5-10°C and, maintaining the temperature at this level, stirred for 1 hour, then filtered, the filter cake was washed with ethyl acetate (5 ml); the filter cake was dried in an oven under vacuum at 50°C for 16 hours; the result was 2.3 g of a pale yellow to off-white solid with an HPLC purity of 99.6%. No impurities containing >0.1% were detected. The total yield at stages 2 and 3 was 42.7%.

MS-ESI and 1H NMR data correspond to example 21.

Example 27 Preparation of a compound of formula I.

THF (120 ml, 12 volumes), (R)-lactamide (13.2 g, 4.0 eq.) and Et ₃ O-BF ₄ (27.8 g, 4.0 eq.), the system was stirred; then the materials in flask No. 1 were mixed under a nitrogen atmosphere and left for later use; to another 500 ml three-neck flask (No. 2) was added ethanol (140 ml, 14 volumes) and the concentrated residue (containing approximately 10 g of compound of formula II) obtained from post-treatment 3 in Example 14; the system was heated to a temperature of 45±2°C in a nitrogen atmosphere; then the materials from three-neck flask No. 1 were added dropwise to three-neck flask No. 2 in one session; to carry out the reaction, the system was maintained at a temperature of 45±5°C (internal temperature 45°C) under a nitrogen atmosphere for 6 hours; then the system was cooled to room temperature and filtered, the filter cake was washed with THF (20 ml), water (20 ml, 2 volumes) was added to the filtrate; then the filtrate was concentrated on a rotary evaporator to a volume of 20-40 ml (2-4 volumes), the concentrated residue was subjected to replacement of the solvent with ethyl acetate (50 mlx2) and concentrated to a volume of 20-40 ml (2-4 volumes), water was added to the concentrated residue (90 ml, 10 parts by volume), the internal temperature of the system was controlled within 20-25°C, a solution of 12 M HCl (8.0 g) was used to adjust the pH of the system to 1-2, then activated carbon was added to the system (1. 0 g) and the system was stirred at room temperature for 2 hours, then filtered, the filter cake was washed with water (20 ml) and a solution of 1 M HCl (20 ml); the combined filtrate was extracted with ethyl acetate (50 mlx2), the organic phase was separated, then the internal temperature of the system was controlled between 20-25°C and the pH was adjusted to 9-10 with a saturated solution of potassium carbonate (28 g); then the internal temperature of the system was controlled within 15-20°C, the system was stirred for 1 hour and filtered, the filter cake was washed with water (20 ml), then it was triturated in an aqueous-acetone solution (100 ml, V/V=1:1 ) for 1 hour, the system was filtered and the filter cake was washed with an aqueous-acetone solution (10 ml, V/V=1:1); The filter cake was blow dried at 50° C. for 16 hours to obtain 15.0 g of a pale gray solid with a purity of 92.4%.

Purification of a compound of formula I.

The resulting solid (10 g) was placed in a flask and ethanol (100 ml) was added; the system was heated to 50-60°C and stirred at 50-60°C for 30 minutes, the materials were mostly dissolved and the solution became clear; Silica gel (20.0 g, 2X) was then added to the system, the system was concentrated to dryness using a rotary evaporator, and left for subsequent use; the crude compound of formula I, mixed with silica gel, was passed through a column of silica gel (100 g, 10X), the column was eluted with ethyl acetate until the former practically disappeared (about 1.5 l of eluent), then eluted with a mixed solution of ethyl acetate and ethanol (V _EA : V _EtOH =100:1-30:1, first 0.5 l V _EA :V _EtOH =100:1, then 1.5 l V _EA :V _EtOH =30:1); the components were checked by TLC and the components containing the product were collected; the collected components were concentrated by rotary evaporation to a volume of 50 ml (5 volume

- 20 045390 parts), after which the solvent was replaced with ethyl acetate (50 mlx2), bringing to a volume of 50 ml (5 parts by volume); then the system was cooled to 5-10°C and stirred for 2 hours, maintaining the temperature at 5-10°C, after which it was filtered, the filter cake was washed with ethyl acetate (5 ml); the mother liquor was concentrated to dryness, resulting in 0.6 g of product; the filter cake was dried in an oven under vacuum at 50°C for 16 hours; the result was 4.6 g of an off-white solid with a purity of 98.1%. The total yield at stages 2 and 3 was 57.6%.

MS-ESI and 1H NMR data correspond to example 21.

Example 28 Preparation of a compound of formula I.

THF (660 ml, 12 volumes), (R)-lactamide (72 g, 4.0 eq.) and Et ₃ O-BF ₄ (153.5 g, 4.0 eq.) were added to a 1000 ml three-neck flask (No. 1). 0 eq.), the system was stirred; then the materials in flask No. 1 were mixed under a nitrogen atmosphere and left for later use; the compound of formula II in THF obtained in example 16 (127.3 g, 55 g of the compound of formula II according to calculations) was placed in a separate flask, the solvent in the system was replaced with ethanol (280 mlx2) and concentrated to 120-180 ml; the concentrated residue was transferred to another 2000 ml three-neck flask (No. 2) and ethanol (770 ml, 14 parts by volume) was added to it; the system was heated to a temperature of 45-50°C (internal temperature); then the materials from three-neck flask No. 1 were added dropwise to three-neck flask No. 2 for 1 hour; to carry out the reaction, the system was maintained at a temperature of 45-50°C (mostly about 45°C) under a nitrogen atmosphere for 3 hours; then the system was cooled to room temperature and filtered, the filter cake was washed with THF (60 ml), water (110 ml, 2 volumes) was added to the filtrate; then the filtrate was concentrated on a rotary evaporator to a volume of 110-220 ml (2-4 volumes), the concentrated residue was subjected to replacement of the solvent with ethyl acetate (280 mlx2) and concentrated to a volume of 110-220 ml (2-4 volumes), water was added to the concentrated residue (550 ml, 10 parts by volume), the internal temperature of the system was controlled within 20-25 ° C, a solution of 12 M HCl (35 g) was used to adjust the pH of the system to 1-2, then activated carbon (5.5 g) was added to the system ) and the system was stirred at room temperature for 2 hours, then filtered, the filter cake was washed with water (110 ml) and a solution of 1 M HCl (110 ml); the combined filtrate was extracted with ethyl acetate (280 mlx2), the organic phase was separated, then the internal temperature of the system was controlled between 20-25°C, and the pH was adjusted to 9-10 with 50% potassium carbonate solution (180 g); then the internal temperature of the system was controlled within 15-20°C, the system was stirred for 2 hours and filtered, the filter cake was washed with water (110 ml), then the filter cake was triturated in an aqueous-acetone solution (550 ml, V/V=1: 1) for 2 hours, the system was filtered and the filter cake was washed with an aqueous-acetone solution (110 ml, V/V=1:1); The filter cake was blown dried at 50° C. for 16 hours to obtain 80 g of a pale gray solid with an HPLC purity of 96.4%, a content of 50.32% and a yield of 61.1. %.

Purification of a compound of formula I.

The resulting solid (80 g), silica gel (160 g, 2X) were placed in a flask and ethanol (800 ml) was added; the system was heated to 50-60°C and stirred at 50-60°C for 30 minutes, then the system was concentrated to dryness on a rotary evaporator and left for subsequent use; the crude compound of formula I, mixed with silica gel, was passed through a column of silica gel (800 g, 10X), the column was eluted with ethyl acetate until the former impurities had practically disappeared, then the column was eluted with a mixed solution of ethyl acetate and ethanol (V _EA : V _EtOH = 100:1-30:1); components were checked by TLC and components were selected that showed a virtual absence of impurities by TLC; the selected components were concentrated on a rotary evaporator to ~200 ml; the concentrated residue was twice subjected to replacing the solvent with methanol (bringing the volume to ~200 ml); then the concentrated residue was subjected to solvent replacement with MTBE twice (bringing the volume to ~200 ml); MTBE (~300 ml) was added to the concentrated residue; the system was refluxed and the system was left to boil for 1 hour; then the system was cooled to 5-10°C, stirred at a temperature in the range of 5-10°C for 2 hours; filtered and the filter cake was washed with MTBE (30 ml); the filter cake was dried in an oven under vacuum at 50°C for 16 hours; the result was 39 g of a pale yellow solid with an HPLC purity of 99.7% and no impurities >0.1%.

MS-ESI and 1H NMR data correspond to example 21.

Example 29 Preparation of a compound of formula I.

THF (11 kg, 12 volumes), (R)-lactamide (1.64 kg, 5.0 eq) and Et ₃ O-BF ₄ (3.50 kg, 5) were added to a 100 L reactor (No. 1). ,0 eq.), the system was stirred; the materials in reactor No. 1 were then mixed under a nitrogen atmosphere and left for later use; the compound of formula II in THF (5.4 kg) obtained in example 19 was placed in a separate flask, the solvent in the system was changed twice to ethanol (4.2 kgx2), then ethanol was added to the system (11.2 kg, 14 parts by volume ) and the system was transferred to another reactor with a volume of 100 l (No. 2); the system was heated in a nitrogen atmosphere to a temperature

- 21 045390

45-50°C (internal temperature); then materials from reactor No. 1 were added dropwise to reactor No. 2 over 1 hour; to carry out the reaction, the system was maintained at a temperature of 45-50°C in a nitrogen atmosphere for 3 hours; then THF (1.0 kg, 12 volumes), (R)-lactamide (0.16 kg, 0.50 eq.) and Et ₃ O-BF ₄ (0.35 kg, 0 .50 eq.) and stirred until dissolved; the resulting solution was added dropwise to reactor No. 2, the reaction was continued for another 2 hours while maintaining the same temperature; then the system was cooled to room temperature and filtered, the filter cake was washed with THF (1.0 kg), water (2 kg, 2 volumes) was added to the filtrate; then the filtrate was concentrated on a rotary evaporator to a volume of 3 l (2-4 volumes), the concentrated residue was subjected to replacing the solvent with ethyl acetate (4 kgx2) to bring it to a volume of 3 l (2-4 volumes), water (10 kg, 10 parts by volume), the internal temperature of the system was controlled within 20-25°C, a solution of 12 M HCl was used to adjust the pH of the system to 1-2, then activated carbon (0.1 kg) was added to the system and the system was stirred at room temperature in for 2 hours, then filtered, the filter cake was washed with water (2 kg) and a solution of 1 M HCl (2 kg); the combined filtrate was extracted with ethyl acetate (3.90 kgx2), the organic phase was separated, then the internal temperature of the system was controlled between 20-25°C and the pH was adjusted to 9-10 with 50% potassium carbonate solution; then the internal temperature of the system was controlled within 15-20°C, the system was stirred for 2 hours and filtered, the filter cake was washed with water (2 kg), then the filter cake was triturated in aqueous acetone solution (8.9 kg, V/V= 1:1) at 25-30°C for 2 hours, then the system was cooled to 5-10°C, triturated for 2 hours and then filtered, the filter cake was washed with an aqueous-acetone solution (1.86 kg, V/V =1:2); the filter cake was dried under vacuum at 50-55°C, resulting in 1.3 kg of solid with a major content of 39.9% and an HPLC purity of 97.1%.

Purification of a compound of formula I.

The above solid (1.3 kg) was charged into a spinning flask and ethanol (6.50 kg) was added; the system was heated to 45-55°C and stirred for 30 minutes at 45-55°C, during which time the materials were mainly dissolved and a clear solution was obtained; then silica gel (2.60 kg) was added to the rotating flask, the flask was placed in a water bath with a temperature of 50-60°C, the system was concentrated under reduced pressure until the liquid fraction was almost completely distilled off; then the solvent in the system was replaced with ethyl acetate (6.50 kg) and the system was concentrated until the liquid fraction was distilled off and left for subsequent use; then the chromatography column was prepared and washed, silica gel (13.00 kg, 200-300 mesh) was loaded into the column, after which the silica gel column was packed with ethyl acetate (26.0 kg); prepared samples were loaded into a column with silica gel, the samples were placed evenly; the column was eluted with ethyl acetate (52.0 kg), after which the column was eluted with a mixed solution of ethyl acetate and ethanol in the ratio ^V EA ^{: V} EtOH = ^{100: 1} (65.6 kg), a mixed solution of ethyl acetate and ethanol in the ratio V _EA : V _EtOH = 80:1 (64.7 kg), mixed solution of ethyl acetate and ethanol in the ratio _VEA :V _EtOH = 50:1 (123.1 kg), mixed solution of ethyl acetate and ethanol in the ratio _VEA :V _EtOH = 30:1 ( 64.8 kg); components were collected with a purity of >99.0% by HPLC; components with a purity of >99.0% by HPLC were pumped into the reactor and concentrated to a minimum stirring volume; 4.0 kg of methanol was pumped into the reactor, concentrated to a minimum mixing volume; then 6.50 kgx5 MTBE was pumped into the reactor, the system was concentrated to a minimum stirred volume; then 6.50 kg of MTBE was pumped into the reactor; the system was heated at reflux and the system was left to reflux for 1 hour; then the system was cooled to 5-10°C and stirred for 2 hours at 5-10°C, then filtered, and the filter cake was washed with MTBE (1.3 kg); as a result, 260 g of wet product was obtained, the filter cake was dried in a vacuum drying oven at 50-55°C for 16 hours; after drying, 0.250 kg of product was obtained with an HPLC purity of 100.0%.

MS-ESI and 1H NMR data correspond to example 21.

In all of the following examples describing the preparation of Crystalline Form 1 of the compound of Formula I, Crystalline Form A hydrochloride of the compound of Formula I, Crystalline Form B of the sulfate of the compound of Formula I, Crystalline Form C of the hydrobromide of the compound of Formula I, and Crystalline Form D of the phosphate of the compound of Formula I were used as the starting material compound of formula I.

Preparation of crystalline form 1 of the compound of formula I.

Example 30.

The compound of formula I (172.6 g) and acetone (2705 g, ~3500 ml) were added to a 5000 ml three-neck flask; the system was heated to 50-60°C and stirred for 2 hours at a temperature of 50-60°C; then the system was cooled to room temperature (25-30°C); stirred at room temperature (25-30°C) for 24 hours; after this, the system was distilled under reduced pressure, the volume of liquid in the flask during distillation decreased to approximately 0.8-0.9 l; this liquid was cooled to 15-25°C, approximately 4.3 kg of purified water was added to it; the system was stirred at room temperature for 2 hours;

- 22 045390 then cooled to 5-10°C, then stirred for 2 hours, filtered with suction, the filter cake was washed with water, then the filter cake was dried in a blower at 50-55°C for 16 hours; the result was 158.8 g of an almost white solid with a yield of 92.0%. This nearly white solid was tested to represent crystalline form 1 of the compound of formula I. An XRPD diffraction pattern, DSC thermogram, TGA thermogram, and DSP isotherm plot of crystalline form 1 are shown in FIG. 1, 2A, 3 and 4 respectively.

Example 31.

The compound of formula I (100 mg) was dissolved in acetone (2 ml) at a temperature in the range of 50-5 5 °C to obtain a clear solution, the said solution was cooled to room temperature and stirred for about 16 hours, then 2 ml of water was added, system stirred at room temperature for 2 hours, cooled to 10-15°C, stirred for 2 hours, filtered and collected the solid. This solid was tested as crystalline form 1 of the compound of formula I. The XRPD diffraction pattern of the crystalline form corresponds to the diffraction pattern shown in FIG. 1.

Example 32.

The compound of formula I (200 mg) was dissolved in 9 ml of methanol/water (9:1) at room temperature to obtain a clear solution, to which 5 mg of seed crystals of crystalline form 1 of the compound of formula I were added, a solid precipitated, then the system was stirred overnight at room temperature, water (18 ml) was added, after which the system was stirred at room temperature for another 4 hours, cooled to 5-10°C and stirred for 1 hour, filtered with suction, dried under vacuum (at about 50° C.), resulting in 180 mg of an almost white solid. This almost white solid was tested to represent crystalline form 1 of the compound of formula I. The XRPD diffraction pattern of the crystalline form corresponds to the diffraction pattern shown in FIG. 1.

Example 33.

The same crystallization method was used as in Example 32. To obtain crystalline form 1 of the compound of formula I, the crystallization solvent was replaced by a mixture of acetone/water (4:1). The XRPD diffraction pattern of the crystalline form corresponds to the diffraction pattern shown in FIG. 1.

Crystalline Form 1 of the compound of formula I can also be prepared by the following method.

Suspension and crystallization at room temperature.

Example 34.

Take a sample of approximately 20 mg of the compound of formula I and place it in a glass vial, add a suitable amount of THF and the vial is subjected to ultrasonic treatment for 5 minutes, resulting in a suspension, then the vial with the sample is wrapped in staniol to protect it from light and placed on laboratory rotator Labquaker, where they rotated 360° at room temperature; a sample of the suspension was centrifuged on the 10th day, the solid residue formed at the bottom was collected, and the solvent was evaporated to dryness, resulting in an almost white solid. This almost white solid was tested to represent crystalline form 1 of the compound of formula I. The XRPD diffraction pattern of the crystalline form corresponds to the diffraction pattern shown in FIG. 1.

Examples 35-40.

The same crystallization method was used as in Example 34. To obtain crystalline form 1 of the compound of formula I, the crystallization solvent was replaced by methyl tert-butyl ether, water, acetone, isopropanol, dichloromethane and ethanol. The results are presented in table. 6 below.

Table 6

Example No. Crystallization solvent Crystalline form of the compound of formula 1 35 Methyl tert-butyl ether Crystal form 1 36 Water Crystal form 1 37 Acetone Crystal form 1 38 Isopropanol Crystal form 1 39 Dichloromethane Crystal form 1 40 Ethanol Crystal form 1

The XRPD diffraction pattern of crystalline form 1 of the compound of formula I obtained in Example 23045390 35-40 corresponds to FIG. 1.

Suspension and crystallization at 50°C.

Example 41.

Take a sample of approximately 20 mg of the compound of formula I and place it in a glass vial, add a suitable amount of THF, and the vial is subjected to ultrasonication for 5 minutes, resulting in a suspension, then the vial containing the sample is wrapped in staniol to protect it from light and placed to a shaker-incubator with a constantly maintained temperature of 50°C for triturating the suspension; a sample of the suspension was centrifuged on the 10th day, the solid residue formed at the bottom was collected, and the solvent was evaporated to dryness, resulting in an almost white solid. This almost white solid was tested to represent crystalline form 1 of the compound of formula I. The XRPD diffraction pattern of the crystalline form corresponds to the diffraction pattern shown in FIG. 1.

Examples 42-45.

The same crystallization method was used as in Example 41. To obtain crystalline form 1 of the compound of formula I, the crystallization solvent was replaced by methyl tert-butyl ether, water, acetone and isopropanol. The results are presented in table. 7 below.

Table 7

Example No. Crystallization solvent Crystalline form of the compound of formula 1 42 Methyl tert-butyl ether Crystal form 1 43 Water Crystal form 1 44 Acetone Crystal form 1 45 Isopropanol Crystal form 1

The XRPD diffraction pattern of crystalline Form 1 of the compound of Formula I obtained in Examples 42-45 corresponds to FIG. 1.

Example 46.

The same crystallization method was used as in Example 41. To obtain crystalline form 1 of the compound of formula I, the crystallization solvent was replaced with methanol. XRPD crystal form corresponds to Fig. 1, indicating that this is crystalline form 1 of the compound of formula I. However, the DSC of the crystalline form shows an additional peak at approximately 151°C, as shown in FIG. 2B.

Heating a saturated solution - rapid cooling and crystallization

Example 47.

A sample of approximately 30 mg of the compound of formula I was taken and placed in a vial, a suitable amount of acetone was added thereto, the vial with the sample was placed on a magnetic stirrer and the temperature of the water bath was controlled within 50 ± 2°C, the rotation speed of the stirrer was set to 200 rpm , the sample was heated to accelerate dissolution, the temperature was maintained for another 15 min after the sample dissolved. The resulting supersaturated solution was filtered through a 0.45 µm membrane while hot, successive filtrates were transferred to a new vial, after which the vial was immediately placed in a freezer at -20°C overnight. The system containing the precipitated solids was then centrifuged and the solids were collected. The solvent was evaporated to dryness naturally, resulting in an almost white solid. This solid was tested as crystalline form 1 of the compound of formula I. The XRPD diffraction pattern of the crystalline form corresponds to the diffraction pattern shown in FIG. 1.

Example 48.

The same crystallization method was used as in Example 47. To obtain crystalline form 1 of the compound of formula I, the crystallization solvent was replaced with THF. The XRPD diffraction pattern of the resulting crystalline form 1 of the compound of formula I corresponds to FIG. 1.

Heating a saturated solution - slow cooling and crystallization.

Example 49.

A sample of approximately 30 mg of the compound of formula I was taken and placed in a vial, a suitable amount of acetone was added thereto, the vial with the sample was placed on a magnetic stirrer and the temperature of the water bath was controlled within 50 ± 2°C, the rotation speed of the stirrer was set to 200 rpm , the sample was heated to accelerate dissolution, the temperature was maintained for another 15 min after the sample dissolved. The resulting supersaturated solution was filtered through a membrane with pores of 0.45 μm in a hot state, successive filtrates were transferred to a new bottle, which was slowly cooled to room temperature at a rate of 6°C/h, placed in a refrigerator and kept

- 24 045390 lived in it for the next 24 hours, i.e. approximately 24 hours (at 2-8°C). The system containing the solvent and precipitated solids was then centrifuged and the solid phase was collected. The solvent was evaporated to dryness naturally, resulting in an almost white solid. This almost white solid was tested to represent crystalline form 1 of the compound of formula I. The XRPD diffraction pattern of the crystalline form corresponds to the diffraction pattern shown in FIG. 1.

Example 50.

The same crystallization method was used as in Example 49. To obtain crystalline form 1 of the compound of formula I, the crystallization solvent was replaced with dichloromethane. The XRPD diffraction pattern of the resulting crystalline form of the compound of formula I corresponds to FIG. 1.

Antisolvent crystallization.

Examples 51-62.

A sample of approximately 30 mg of the compound of formula I was taken and placed in a vial, a certain volume of a good solvent was added thereto, and the vial with the sample was subjected to ultrasonic treatment at room temperature to ensure uniform dispersion of the sample. If the solution turned out to be transparent, more solid sample was added to it and ultrasonic treatment was continued to facilitate dissolution and obtain a supersaturated solution of the sample in a good solvent. The solution was filtered through a membrane with a pore size of 0.45 μm, the first portion of the filtrate was separated, the subsequent filtrate was transferred to a new bottle, an antisolvent was added there with constant stirring with a volume 10 times greater than the volume of a good solvent, and stirring was maintained at a speed of 50 rpm. min, then the solvent system with the precipitated solid phase was centrifuged and an almost white solid was obtained. The nearly white solid obtained in Examples 51-62 was tested to represent crystalline form 1 of the compound of formula I. The XRPD diffraction pattern of the crystalline form corresponds to the diffraction pattern shown in FIG. 1. The results obtained are presented in table. 8 below.

Table 8

Example No. Solvent for crystallization (good solvent antisolvent) Crystalline form of the compound of formula I 51 Methanol - water (1:10) Crystal form 1 52 Ethanol - water (1:10) Crystal form 1 53 THF - methyl tert-butyl ether (1:10) Crystal form 1 54 Ethanol methyl tert-butyl ether (1:10) Crystal form 1 55 Methanol - methyl tert-butyl ether (1:10) Crystal form 1 56 Acetone - methyl tert-butyl ether (1:10) Crystal form 1 57 Isopropanol - methyl tert-butyl ether (1:10) Crystal form 1 58 THF - dichloromethane (1:10) Crystal form 1 59 Ethanol - dichloromethane (1:10) Crystal form 1

- 25 045390

60 Methanol - dichloromethane (1:10) Crystal form 1 61 Acetone - dichloromethane (1:10) Crystal form 1 62 Isopropanol - dichloromethane (1:10) Crystal form 1

Preparation of crystalline Form A hydrochloride of the compound of formula I.

Example 63.

A sample of approximately 200 mg of a compound of formula I was taken and placed in a vial, 4 ml of ethanol was added thereto, the vial was subjected to ultrasonic treatment and heating until the sample was completely dissolved, then the vial with the sample was placed on a magnetic stirrer, the resulting solution was constantly stirred and, while stirring, 1.32 ml of a solution of hydrochloric acid in ethanol was slowly added dropwise (the concentration of hydrochloric acid in ethanol was 50 mg/ml), a white precipitate formed, the bottle was tightly capped at room temperature and stirred for 1 day, then the reaction suspension centrifuged and the collected solid was dried under vacuum at 40° C. overnight to obtain the solid hydrochloride of the compound of formula I. The solid was tested to represent the hydrochloride crystalline Form A of the compound of formula I. An XRPD diffraction pattern and a DSC thermogram of the crystalline form are shown in fig. 5 and 6.

Example 64.

A sample of approximately 50 mg of a compound of formula I was taken and placed in a vial, 1 ml of ethanol was added thereto, the vial was subjected to ultrasonic treatment and heating until the sample was completely dissolved, then the vial with the sample was placed on a magnetic stirrer, the resulting solution was constantly stirred and, while stirring, 0.33 ml of a solution of hydrochloric acid in ethanol was slowly added dropwise (the concentration of hydrochloric acid in ethanol was 50 mg/ml), a white precipitate formed, the bottle was tightly capped at room temperature and stirred for 1 day, then the reaction suspension centrifuged and the collected solid was dried under vacuum at 40° C. overnight to obtain the solid hydrochloride of the compound of formula I. The solid was tested to represent the hydrochloride crystalline Form A of the compound of formula I. The XRPD diffraction pattern of the crystalline form corresponds to the diffraction pattern shown in fig. 5.

Preparation of sulfate crystalline form B of the compound of formula I.

Example 65.

A sample of approximately 200 mg of a compound of formula I was taken and placed in a vial, 4 ml of ethanol was added thereto, the vial was subjected to ultrasonic treatment and heating until the sample was completely dissolved, then the vial with the sample was placed on a magnetic stirrer, the resulting solution was constantly stirred and, while stirring, 1.4 ml of a solution of sulfuric acid in ethanol was slowly added dropwise (the concentration of sulfuric acid in ethanol was 50 mg/ml), a white precipitate formed, the bottle was tightly capped at room temperature and stirred for 1 day, then the reaction suspension centrifuged and the collected solid was dried under vacuum at 40° C. overnight to obtain a solid sulfate of the compound of formula I. The solid was tested to be crystalline form B of the sulfate of the compound of formula I. XRPD diffraction pattern, DSC thermogram, TGA thermogram and The DSP isotherm graph is shown in Fig. 7-10.

Example 66.

A sample of approximately 50 mg of a compound of formula I was taken and placed in a vial, 1 ml of ethanol was added thereto, the vial was subjected to ultrasonic treatment and heating until the sample was completely dissolved, then the vial with the sample was placed on a magnetic stirrer, the resulting solution was constantly stirred and, while stirring, 0.35 ml of a solution of sulfuric acid in ethanol was slowly added dropwise to it (the concentration of sulfuric acid in ethanol was 50 mg/ml), a white precipitate formed, the bottle was tightly capped at room temperature and stirred for 1 day, then the reaction suspension centrifuged and the collected solid was dried under vacuum at 40° C. overnight to obtain a solid sulfate of the compound of formula I. The solid was tested to be crystalline form B of the sulfate of the compound of formula I. The XRPD diffraction pattern of the resulting substance corresponds to the diffraction pattern shown in fig. 7.

Preparation of the hydrobromide crystalline form C of the compound of formula I.

Example 67.

A sample of approximately 200 mg of the compound of formula I was taken and placed in a vial, 4 ml of ethanol was added thereto, the vial was subjected to ultrasonic treatment and heated until the solution was completely dissolved.

-26045390 sample, then the bottle with the sample was placed on a magnetic stirrer, the resulting solution was constantly stirred, and while stirring, 2.4 ml of a solution of hydrobromic acid in ethanol was slowly added dropwise (the concentration of hydrobromic acid in ethanol was 50 mg/ml), at This formed a white precipitate, the vial was tightly capped at room temperature and stirred for 1 day, then the reaction suspension was centrifuged and the collected solid was dried under vacuum at 40°C overnight, resulting in the solid hydrobromide of the compound of formula I. Based on the results The test solid is the hydrobromide crystalline form C of the compound of formula I.

The XRPD diffraction pattern and DSC thermogram of the resulting substance are shown in FIG. 11 and 12.

Example 68.

A sample of approximately 50 mg of a compound of formula I was taken and placed in a vial, 1 ml of ethanol was added thereto, the vial was subjected to ultrasonic treatment and heating until the sample was completely dissolved, then the vial with the sample was placed on a magnetic stirrer, the resulting solution was constantly stirred and, while stirring, 0.60 ml of a solution of hydrobromic acid in ethanol was slowly added dropwise (the concentration of hydrobromic acid in ethanol was 50 mg/ml), a white precipitate formed, the bottle was tightly capped at room temperature and stirred for 1 day, then the reaction suspension centrifuged and the collected solid was dried under vacuum at 40° C. overnight to obtain the solid hydrobromide of the compound of formula I. The solid was tested to represent the hydrobromide crystalline form C of the compound of formula I. The XRPD diffraction pattern of the resulting crystalline form corresponds to the diffraction pattern shown in fig. eleven.

Preparation of crystalline phosphate form D of the compound of formula I.

Example 69.

A sample of approximately 200 mg of a compound of formula I was taken and placed in a vial, 4 ml of ethanol was added thereto, the vial was subjected to ultrasonic treatment and heating until the sample was completely dissolved, then the vial with the sample was placed on a magnetic stirrer, the resulting solution was constantly stirred and, while stirring, 1.56 ml of a solution of phosphoric acid in ethanol was slowly added dropwise to it (the concentration of phosphoric acid in ethanol was 50 mg/ml), a white precipitate formed, the bottle was tightly capped at room temperature and stirred for 1 day, then the reaction suspension centrifuged and the collected solid was transferred to another vial, 3-4 ml of mixed solvent acetone/water (9/1, V/V) was added thereto, stirred with a magnetic stirrer at room temperature overnight, then the mixture was centrifuged, the solid was separated and dried under vacuum at 40°C overnight to obtain the solid phosphate of the compound of formula I. The solid was tested to be the phosphate crystalline form D of the compound of formula I. XRPD diffraction pattern, DSC thermogram, TGA thermogram and DSP isotherm plot of the resulting substances are shown in Fig. 13-16.

Example 70.

A sample of approximately 50 mg of a compound of formula I was taken and placed in a vial, 1 ml of ethanol was added thereto, the vial was subjected to ultrasonic treatment and heating until the sample was completely dissolved, then the vial with the sample was placed on a magnetic stirrer, the resulting solution was constantly stirred and, while stirring, 0.39 ml of a solution of phosphoric acid in ethanol was slowly added dropwise to it (the concentration of phosphoric acid in ethanol was 50 mg/ml), a white precipitate formed, the bottle was tightly capped at room temperature and stirred for 1 day, then the reaction suspension centrifuged and the collected solid was transferred to another vial, 0.75-1 ml of mixed solvent acetone/water (9/1, V/V) was added thereto, stirred with a magnetic stirrer at room temperature overnight, then the mixture was centrifuged, solid the substance was separated and dried under vacuum at 40° C. overnight to obtain the solid phosphate of the compound of formula I. The solid was tested to be the phosphate crystalline form D of the compound of formula I. The XRPD diffraction pattern of the resulting crystalline form corresponds to the diffraction pattern shown in FIG. . 13.

experimental part

Experimental example 1. Characteristics of the solid.

DSC, TGA and EAF tests were performed on the hydrochloride crystalline Form A of the compound of Formula I, the sulfate crystalline Form B of the compound of Formula I, the hydrobromide crystalline Form C of the compound of Formula I and the phosphate crystalline Form D of the compound of Formula I.

The results of DSC tests showed that the sulfate crystalline Form B of the compound of formula I (Fig. 8) and the phosphate crystalline Form D of the compound of Formula I (Fig. 14) have single and elevated melting points, indicating relatively good thermal stability of the crystalline forms. The thermodynamic behavior of the hydrochloride crystal form A of the compound of formula I (Fig. 6) and the hydrobromide crystal form C of the compound of formula I (Fig. 12) is more

- 27 045390 complex, and their crystallinity is lower. Thus, further characterization by DSP and

TGA was performed only on the sulfate crystalline form B of the compound of formula I and the phosphate crystalline form D of the compound of formula I.

The results of the EAF tests showed that the sulfate crystalline form B of the compound of formula I (Fig. 10) and the phosphate crystalline form D of the compound of formula I (Fig. 16) have a certain hygroscopicity. The increase in mass due to moisture absorption when the ambient relative humidity changes from 0% RH to 95% RH for these forms is 9.4 and 4.7%, respectively. No significant changes were found in the crystalline forms of these two salts before and after EAF testing.

The results of the TGA test showed that the phosphate crystalline form D of the compound of formula I (FIG. 15) had a mass loss of 2.862% over the temperature range from 44 to 120°C.

Based on the results obtained for the crystalline forms of these four salts, their comparison with crystalline form 1 of the compound of formula I is presented in table. 9 below.

Table 9

Crystalline form 1 of the compound of formula I Crystalline form A hydrochloride of the compound of formula I Crystalline form B sulfate of the compound of formula I Crystalline form C hydrobromide of the compound of formula I Crystalline phosphate form D of the compound of formula I Crystallinity Great Fine Great Fine Great Initial 161°С No 199°С 145°С 207°C melting temperature apparent melting point TGA -5.4% (25-133°C) - - - -2.9% (44-120°C) Gyroscopicity (moisture absorption at 95% relative humidity) 5.2% - 9.4% - 4.7%

means the test was not performed.

It can be concluded that the hydrochloride crystalline form A of the compound of formula I and the hydrobromide crystalline form C of the compound of formula I have wide melting ranges and their melting points are not obvious; The hygroscopicity of the sulfate crystal form B of the compound of formula I is higher than that of the phosphate crystal form D of the compound of formula I. The phosphate crystal form 1 of the compound of formula I and the phosphate crystal form D of the compound of formula I have better crystallinity, single melting points and relatively low hygroscopicity.

Experimental Example 2: Solubility Test.

We took 4 weighed samples of crystalline form 1 of the compound of formula I and crystalline form D of the phosphate of the compound of formula I, placed them in transparent glass bottles with a volume of 4 ml and added 1 ml of water, simulated gastric juice (SGF), and simulated intestinal juice on an empty stomach, respectively. (FaSSIF) and simulated intestinal juice after a meal (FeSSIF), a sample suspension was obtained and quickly placed on a shaker (at 37°C, 200 rpm), shaken on the shaker, after 5 minutes the samples were examined, the number of samples or media was replenished, so to obtain a moderate suspension, samples were collected at 30 min, 2, 4, and 24 h, respectively, centrifuged for 10 min at 12,000 rpm, and the supernatants were collected, diluted appropriately, and tested by high-performance liquid chromatography. Chromatography conditions are presented in table. 10.

-28045390

Table 10

Conditions for solubility testing by high performance liquid chromatography

Device Agilent 1200 DAD H PLC System Chromatographic column Waters XBridge Shield RP18 4.6x150 mm, 3.5 µm Mobile phase A: 0.05% aqueous phosphoric acid solution B: Acetonitrile Column temperature 30°C Detector DAD Detection wavelength 232 nm Injected sample volume 10 µl Column flow rate 1.0 ml/min Chromatography time 10 min Data collection time 10 min Elution procedure Time (min) A (%) V (%) 0.0 95 5 6.0 70 thirty 7.0 5 95 7.1 95 5 10 95 5

Sample concentration values were calculated using the external standard method. The results of the tests are presented in table. eleven.

Table 11

Sample Wednesday Solubility (based on compound of formula I, mg/ml) 30 min 2h 4 hours 24 hours Crystal Form 1 of Compound of Formula I Water 0.16 0.18 0.16 0.18 Artificial Gastric Fluid (SGF) 16.52 16.71 16.88 16.42 Artificial fasting intestinal juice (FaSSIF) 0.17 0.19 0.16 0.17 Simulated intestinal juice after eating (FeSSIF) 0.26 0.23 0.22 0.24 Phosphate crystalline form D of the compound of formula I Water 1.85 1.78 1.73 1.80 Artificial Gastric Fluid (SGF) 30.22 29.73 29.47 30.18 Simulated fasting intestinal fluid (FaSSIF) 0.17 0.17 0.17 0.16 Simulated intestinal juice after eating (FeSSIF) 0.36 0.22 0.19 0.21

- 29 045390

The results showed no obvious difference in the equilibrium solubilities (after 24 hours) of Formula I crystal Form 1 and Formula I phosphate crystal Form D for FaSSIF and FeSSIF. For water and SGF, the equilibrium solubility values (after 24 hours) for the phosphate crystalline form D of the compound of formula I were 1.8 mg/ml and 30.18 mg/ml, respectively, i.e. respectively, 10 times and 2 times the solubility of crystalline form 1 of the compound of formula I. The solubility values of crystalline form 1 of the compound of formula I and crystalline form D of the phosphate of the compound of formula I according to the present invention correspond to the requirements established for medicinal products.

Experimental Example 3: Stability Test.

Weights of approximately 1 mg of crystalline form 1 of compound of formula I and 1 mg of crystalline form D of phosphate of compound of formula I were placed in a clear glass vial with a volume of 40 ml, then the samples were placed in a chamber for stability studies under accelerated aging conditions (at 40 ° C and relative humidity 75 % in the open state) and at elevated temperatures (at 60°C in the closed state), respectively. In the case of testing open samples, the cap of the vial was removed and the neck of the vial was covered with foil paper with punctures to avoid cross-contamination; In the case of testing closed samples, each vial was tightly and hermetically sealed with a lid. Samples were collected after 1 and 2 weeks, respectively, and after dilution with a diluent (acetonitrile/water, 1:1, v/v), the liquid phases were introduced into the chromatograph and analyzed in accordance with the chromatographic conditions specified in the table. 12, to determine the purity of the samples.

Table 12

Device Agilent 1200 DAD HPLC System Chromatographic column Agilent ZORBAX SB-Phenyl 150*4.6 mm*3.5 mkm Mobile phase A: Aqueous solution of 20 mM ammonium acetate B: Methanol Column temperature 40°C Detector DAD Detection wavelength 220 nm Injected sample volume 5.0 µl Column flow rate 1.0 ml/min Chromatography time 42 min Data collection time 42 min Elution procedure Time (min) A (%) V (%) 0 70 thirty 14 46.5 53.5 thirty 25 75 35 25 75 35.01 70 thirty 42 70 thirty

The purity of the samples was calculated using the area normalization method. The results of the tests are presented in table. 13.

- 30 045390

Table 13

Sample Stability study (purity, area %) 0 day 60°C 40°C/75% rel. humid 1 Week 2 weeks Crystallin's form 1 Week 2 weeks Crystalline form Crystallin form 1 of the compound of formula I 98.7 4 98.73 98.74 No changes 98.71 98.77 No changes Crystalline form D phosphate of the compound of formula I 99.1 3 99.11 99.11 No changes 99.11 99.10 No changes

The results showed that the appearance of Crystal Form 1 of Compound of Formula 1 and Crystalline Form D of Phosphate of Compound of Formula I did not change after 2 weeks and they were still an almost white powder. No significant difference in their purity was observed, and the total content of related substances in them did not increase. XRPD and DSC testing (see Figures 17-20) showed that there was no significant difference in the crystalline forms and initial melting points of the two samples compared to their values on day 0, indicating good 2-week physical and chemical stability crystalline form 1 of the compound of formula I and crystalline form D of the phosphate of the compound of formula I under conditions of elevated temperatures (at 60°C) and under conditions of accelerated aging (at 40°C and 75% relative humidity).

Experimental example 4. Biochemical tests on TYK2.

To carry out biochemical tests on TYK2, a suitable amount of a compound of formula I was weighed.

Testing was performed by Reaction Biology Corp (Malvern, PA) (Anastassiadis et al., Nat. Biotechnol., 2011, 29(11):1039-45). The test procedure is briefly described as follows.

Reagents.

Main working buffer: 20 mM Hepes (pH 7.5), 10 mM MgCl ₂ , 1 mM EGTA, 0.02% Brij35, 0.02 mg/ml BSA, 0.1 mM Na ₃ VO ₄ , 2 mM DTT and 1% DMSO. Accordingly, the required cofactor was added during each kinase reaction.

Procedure for testing kinase reactivity.

1. Prepare and dissolve the intended substrate in freshly prepared basic working buffer.

2. Transfer the required cofactor to the above matrix solution.

3. Transfer the intended kinase to the substrate solution and gently mix.

4. Transfer the compound of formula I in DMSO to the kinase reaction mixture using an Echo 550 acoustic dispensing station (nanolite range), culturing for 20 minutes at room temperature.

5. Add ³³ P-ATP (specific activity: 10 µCi/µL) to the reaction mixture to initiate the reaction.

6. Cultivate at room temperature and carry out the kinase reaction for 2 hours.

7. Applying the reaction mixture to P81 ion exchange paper.

8. Testing kinase activity using the filter binding method.

The test results showed that the compound of formula I is also a potent inhibitor of TYK2, and its IC ₅₀ is less than 10 nM.

One skilled in the art will appreciate that modifications or changes can be made to the present invention based on the instructions provided herein. All such modifications and changes should be considered within the scope of the claims.

-

Claims (5)

CLAIM 1. A method for preparing a compound of formula I, the synthesis scheme of which is as follows: HCI IV III II I where said method includes the following steps: step 1, including adding ethanol, a compound of formula IV, a compound of formula V and DIPEA (diisopropylethylamine) to the reaction vessel, stirring; heating to a temperature of 65-90°C, maintaining the specified temperature and stirring overnight; completion of the reaction and reduction of the system temperature to 15-30°C; adding water dropwise to the system followed by stirring; filtering and washing the filter cake; drying the filter cake to obtain a compound of formula III; step 2, comprising adding tetrahydrofuran (THF), the compound of formula III obtained in step 1, and palladium-carbon to the reaction vessel; purging the system with nitrogen and then hydrogen; maintaining the temperature in the range of 20-35°C and stirring for 16-120 hours at a hydrogen pressure of 0.1-1.0 MPa; filtering the reaction liquid after completion of the reaction and washing the filter cake; combining the filtrates and concentrating to obtain a concentrate of the compound of formula II; step 3, including adding THF, (R)-lactamide and Et ₃ O-BF ₄ to the first reaction vessel, stirring and dissolving for subsequent use; adding said concentrate of the compound of formula II and ethanol to the second reaction vessel and heating the materials in the second vessel to a temperature of 40-85°C; adding materials from the first reaction vessel dropwise to the second reaction vessel and upon completion of addition maintaining the temperature in the range of 40-85°C to allow the mixture of materials to react in the second reaction vessel for 0.5-6 hours; after completion of the reaction, bringing the pH of the system to 1-3, extracting with an organic solvent or organic solvents, removing the organic phase, adjusting the pH of the aqueous phase to 9-10 with an aqueous solution of inorganic alkali, filtering and drying the filter cake to obtain a compound of formula I. 2. The method according to claim 1, in which in said step 1 the volume-mass ratio of ethanol and the compound of formula IV is from 5:1 to 20:1, preferably 10:1; the molar ratio of the compound of formula IV, the compound of formula V and DIPEA is 1:1-1.1:2-3, preferably 1:1.01:2.2; after turning on the stirring, heat the reaction mixture in a nitrogen atmosphere to 65-90°C, preferably to 70-90°C, more preferably to 70-80°C, maintain the specified temperature - 32 045390 and stirred for 5-16 hours, preferably for 10-16 hours; after completion of the reaction, reduce the temperature in the system to 15-25°C; the volume-mass ratio of water added to the system and the compound of formula IV is from 10:1 to 20:1, preferably 15:1; after adding water to the system, stir for 2-6 hours, preferably for 4 hours, at a temperature of 0-30°C, preferably at 5-15°C, more preferably at 5-10°C; wash the filter cake with a solution of ethanol in water, wherein the volume ratio of ethanol and water in the solution is from 1:1 to 1:2, preferably from 1:1.5 to 1:2, and the volume-mass ratio of the ethanol solution and the compound of formula IV is from 2:1 to 10:1, preferably from 2:1 to 5:1, more preferably from 2:1 to 3:1; dry the filter cake under vacuum or blower at a temperature of 45-55°C, preferably 50°C. 3. The method according to claim 1 or 2, in which, in said step 2, the volume-mass ratio of THF to the compound of formula III is from 10:1 to 70:1, preferably from 20:1 to 70:1; palladium on carbon is 5% Pd/C, palladium on carbon with 50% moisture, the weight ratio of palladium on carbon to the compound of formula III is from 0.15:1 to 0.16:1, preferably 0.15:1; maintain the temperature at 25-35°C and stir for 24-96 hours at a hydrogen pressure of 0.5-1.0 MPa; wash the filter cake with THF, wherein the concentrate of the compound of formula II obtained by combining the filtrates and concentrating is the compound of formula II in THF, and the volume-to-mass ratio (ml/g) of THF for washing and the compound of formula II is from 2:1 to 4 :1, preferably from 2:1 to 3:1; preferably replacing THF in a solution of the compound of formula II with ethanol to obtain a compound of formula II in ethanol, wherein the volume/weight ratio (ml/g) of ethanol to compound of formula II is from 2:1 to 5:1, preferably from 2:1 to 4: 1, more preferably 2:1 to 3:1. 4. The method according to any one of claims 1 to 3, in which, in said step 3, the volume-to-mass ratio of THF to concentrate of the compound of formula II is from 6:1 to 12:1; the molar ratio of the concentrate of the compound of formula II, (R)-lactamide and Et ₃ O-BF ₄ is 1:4-5:4-5; the volume/mass ratio of ethanol to concentrate of the compound of formula II is from 10:1 to 16:1, preferably 14:1; after adding the concentrate of the compound of formula II and ethanol to the second reaction vessel, heat the materials in the second reaction vessel under a nitrogen atmosphere to a temperature of 40-85°C, preferably 45-70°C, more preferably 45-50°C; maintain the temperature at 45-70°C, preferably 45-50°C, and allow the materials mixed in the second reaction vessel to react for 2-5 hours, preferably for 3 hours; after completion of the reaction, adjust the pH of the system to 1-3 with hydrochloric acid, which is 1 M or 12 M HCl, preferably 12 M HCl; said aqueous inorganic alkali solution is a saturated sodium carbonate solution or a saturated potassium carbonate solution, preferably a saturated potassium carbonate solution; the organic solvent used to extract the product after completion of the reaction is dichloromethane or ethyl acetate; dry the filter cake under vacuum or blower at a temperature of 50-55°C. 5. A method according to any one of claims 1 to 4, comprising separating and purifying the compound of formula I obtained in said step 3 using column chromatography, where a mixed solution of ethyl acetate and ethanol is used as the eluent. -