WO2025157260A1 - Pharmaceutically acceptable salt of g12d inhibitor compound and crystal form thereof - Google Patents

Abstract

The present disclosure relates to a pharmaceutically acceptable salt of a G12D inhibitor compound and a crystal form thereof. Specifically, provided in the present disclosure are a salt of 2-amino-4-((5S,5aS,6S,9R)-12-((1-((4-(difluoromethylene)piperidin-1-yl)methyl)cyclopropyl)methoxy)-1-fluoro-5-methyl-5a,6,7,8,9,10-hexahydro-5H-4-oxa-3,10a,11,13,14-pentaaza-6,9-methanonaphtho[1,8-ab]cyclohepten-2-yl)-7-fluorobenzo[b]thiophene-3-carbonitrile and a crystal form thereof.

Description

Pharmaceutically acceptable salts of G12D inhibitor compounds and their crystal forms

This application claims the benefit of Chinese Patent Application No. 2024101121666, filed January 26, 2024. This application incorporates the entirety of the aforementioned Chinese Patent Application.

Technical Field

The present disclosure belongs to the field of pharmaceuticals and relates to pharmaceutically acceptable salts of G12D inhibitor compounds and their preparation methods and crystal forms.

Background Art

The KRAS protein lacks traditional small molecule binding sites on its surface and has an extremely high affinity for guanylate, making it extremely difficult to inhibit. Long considered an undruggable drug target, however, given the importance and prevalence of KRAS activation in cancer progression, KRAS has been and remains a highly sought-after target for drug development. As a mutant with widespread and overexpressed expression in various tumors, G12D, the development of inhibitors targeting it, holds significant clinical significance.

WO2024022444 discloses a novel G12D inhibitor compound, 2-amino-4-((5S,5aS,6S,9R)-12-((1-((4-(difluoromethylidene)piperidin-1-yl)methyl)cyclopropyl)methoxy)-1-fluoro-5-methyl-5a,6,7,8,9,10-hexahydro-5H-4-oxa-3,10a,11,13,14-pentaaza-6,9-methylnaphtho[1,8-ab]heptylcyclo-2-yl)-7-fluorobenzo[b]thiophene-3-carbonitrile (Compound A).

Salt formation can improve certain undesirable physicochemical or biological properties of drugs. The development of salts with superior physicochemical or pharmaceutical properties compared to 2-amino-4-((5S,5aS,6S,9R)-12-((1-((4-(difluoromethylidene)piperidin-1-yl)methyl)cyclopropyl)methoxy)-1-fluoro-5-methyl-5a,6,7,8,9,10-hexahydro-5H-4-oxa-3,10a,11,13,14-pentaaza-6,9-methano[1,8-ab]heptylcyclo-2-yl)-7-fluorobenzo[b]thiophene-3-carbonitrile is of great significance. In view of the importance of solid drug crystal forms and their stability in clinical treatment, in-depth research on the pharmaceutically acceptable salt crystal forms of the compound 2-amino-4-((5S,5aS,6S,9R)-12-((1-((4-(difluoromethylidene)piperidin-1-yl)methyl)cyclopropyl)methoxy)-1-fluoro-5-methyl-5a,6,7,8,9,10-hexahydro-5H-4-oxa-3,10a,11,13,14-pentaaza-6,9-methylnaphtho[1,8-ab]heptylcyclo-2-yl)-7-fluorobenzo[b]thiophene-3-carbonitrile is also of great significance for the development of drugs suitable for industrial production and with good biological activity.

Summary of the Invention

In one aspect, the present disclosure provides a pharmaceutically acceptable salt of the compound 2-amino-4-((5S,5aS,6S,9R)-12-((1-((4-(difluoromethylidene)piperidin-1-yl)methyl)cyclopropyl)methoxy)-1-fluoro-5-methyl-5a,6,7,8,9,10-hexahydro-5H-4-oxa-3,10a,11,13,14-pentaaza-6,9-methano[1,8-ab]heptan-2-yl)-7-fluorobenzo[b]thiophene-3-carbonitrile.

In some embodiments, the pharmaceutically acceptable salt is selected from the group consisting of hydrochloride, sulfate, phosphate, L-tartrate, maleate, citrate, L-malate, p-toluenesulfonate, methanesulfonate, benzoate, succinate, and fumarate.

In other embodiments, the chemical ratio of the compound 2-amino-4-((5S,5aS,6S,9R)-12-((1-((4-(difluoromethylidene)piperidin-1-yl)methyl)cyclopropyl)methoxy)-1-fluoro-5-methyl-5a,6,7,8,9,10-hexahydro-5H-4-oxa-3,10a,11,13,14-pentaaza-6,9-methylnaphtho[1,8-ab]heptyl-2-yl)-7-fluorobenzo[b]thiophene-3-carbonitrile to the acid molecule is 1:0.5 to 1:3, including 1:0.5, 1:1, 1:2 or 1:3.

In some embodiments, the chemical ratio of the compound 2-amino-4-((5S,5aS,6S,9R)-12-((1-((4-(difluoromethylidene)piperidin-1-yl)methyl)cyclopropyl)methoxy)-1-fluoro-5-methyl-5a,6,7,8,9,10-hexahydro-5H-4-oxa-3,10a,11,13,14-pentaaza-6,9-methanaptho[1,8-ab]heptyl-2-yl)-7-fluorobenzo[b]thiophene-3-carbonitrile and the acid molecule is 1:1.

Some embodiments provide the hydrochloride salt of the compound 2-amino-4-((5S,5aS,6S,9R)-12-((1-((4-(difluoromethylidene)piperidin-1-yl)methyl)cyclopropyl)methoxy)-1-fluoro-5-methyl-5a,6,7,8,9,10-hexahydro-5H-4-oxa-3,10a,11,13,14-pentaaza-6,9-methanaptho[1,8-ab]heptyl-2-yl)-7-fluorobenzo[b]thiophene-3-carbonitrile, wherein the chemical ratio of the compound to HCl is 1:1 to 1:2.

Some embodiments provide the L-tartrate salt of the compound 2-amino-4-((5S,5aS,6S,9R)-12-((1-((4-(difluoromethylidene)piperidin-1-yl)methyl)cyclopropyl)methoxy)-1-fluoro-5-methyl-5a,6,7,8,9,10-hexahydro-5H-4-oxa-3,10a,11,13,14-pentaaza-6,9-methanaptho[1,8-ab]heptyl-2-yl)-7-fluorobenzo[b]thiophene-3-carbonitrile, wherein the chemical ratio of the compound to L-tartaric acid is 1:1.

Some embodiments provide a maleate salt of the compound 2-amino-4-((5S,5aS,6S,9R)-12-((1-((4-(difluoromethylidene)piperidin-1-yl)methyl)cyclopropyl)methoxy)-1-fluoro-5-methyl-5a,6,7,8,9,10-hexahydro-5H-4-oxa-3,10a,11,13,14-pentaaza-6,9-methanaptho[1,8-ab]heptyl-2-yl)-7-fluorobenzo[b]thiophene-3-carbonitrile, wherein the chemical ratio of the compound to maleic acid is 1:1.

Some embodiments provide a citrate salt of the compound 2-amino-4-((5S,5aS,6S,9R)-12-((1-((4-(difluoromethylidene)piperidin-1-yl)methyl)cyclopropyl)methoxy)-1-fluoro-5-methyl-5a,6,7,8,9,10-hexahydro-5H-4-oxa-3,10a,11,13,14-pentaaza-6,9-methanaptho[1,8-ab]heptyl-2-yl)-7-fluorobenzo[b]thiophene-3-carbonitrile, wherein the chemical ratio of the compound to citric acid is 1:1 to 1:2.

Some embodiments provide a succinate salt of the compound 2-amino-4-((5S,5aS,6S,9R)-12-((1-((4-(difluoromethylidene)piperidin-1-yl)methyl)cyclopropyl)methoxy)-1-fluoro-5-methyl-5a,6,7,8,9,10-hexahydro-5H-4-oxa-3,10a,11,13,14-pentaaza-6,9-methanaptho[1,8-ab]heptyl-2-yl)-7-fluorobenzo[b]thiophene-3-carbonitrile, wherein the chemical ratio of the compound to succinic acid is 1:1.

Some embodiments provide a methanesulfonate salt of the compound 2-amino-4-((5S,5aS,6S,9R)-12-((1-((4-(difluoromethylidene)piperidin-1-yl)methyl)cyclopropyl)methoxy)-1-fluoro-5-methyl-5a,6,7,8,9,10-hexahydro-5H-4-oxa-3,10a,11,13,14-pentaaza-6,9-methanaptho[1,8-ab]heptyl-2-yl)-7-fluorobenzo[b]thiophene-3-carbonitrile, wherein the chemical ratio of the compound to methanesulfonic acid is 1:1 to 1:2.

Some embodiments provide a citrate salt of the compound 2-amino-4-((5S,5aS,6S,9R)-12-((1-((4-(difluoromethylidene)piperidin-1-yl)methyl)cyclopropyl)methoxy)-1-fluoro-5-methyl-5a,6,7,8,9,10-hexahydro-5H-4-oxa-3,10a,11,13,14-pentaaza-6,9-methanaptho[1,8-ab]heptyl-2-yl)-7-fluorobenzo[b]thiophene-3-carbonitrile, wherein the chemical ratio of the compound to citric acid is 1:1 or 1:2.

Another aspect of the present disclosure provides a pharmaceutically acceptable salt of the compound 2-amino-4-((5S,5aS,6S,9R)-12-((1-((4-(difluoromethylidene)piperidin-1-yl)methyl)cyclopropyl)methoxy)-1-fluoro-5-methyl-5a,6,7,8,9,10-hexahydro-5H-4-oxa-3,10a,11,13,14-pentaaza-6,9-methano[1,8-ab]heptan-2-yl)-7-fluorobenzo[b]thiophene-3-carbonitrile The preparation method includes the step of forming a salt of the compound 2-amino-4-((5S, 5aS, 6S, 9R)-12-((1-((4-(difluoromethylidene)piperidin-1-yl)methyl)cyclopropyl)methoxy)-1-fluoro-5-methyl-5a, 6, 7, 8, 9, 10-hexahydro-5H-4-oxa-3, 10a, 11, 13, 14-pentaaza-6, 9-methylnaphtho[1, 8-ab]heptyl-2-yl)-7-fluorobenzo[b]thiophene-3-carbonitrile with an acid.

In some embodiments, the acid used in the salt-forming reaction is selected from hydrochloric acid, sulfuric acid, phosphoric acid, L-tartaric acid, maleic acid, citric acid, L-malic acid, p-toluenesulfonic acid, methanesulfonic acid, benzoic acid, succinic acid and fumaric acid. In some embodiments, the compound 2-amino-4-((5S,5aS,6S,9R)-12-((1-((4-(difluoromethylidene)piperidin-1-yl)methyl)cyclopropyl)methoxy)-1-fluoro-5-methyl-5a,6,7,8,9,10-hexahydro-5H-4-oxa-3,10a,11,13,14-pentaaza-6,9-methylnaphtho[1,8-ab]heptylcyclo-2-yl)-7-fluorobenzo[b]thiophene-3-carbonitrile is reacted with hydrogen chloride/ethanol solution to form a hydrochloride salt.

In some embodiments, the compound 2-amino-4-((5S,5aS,6S,9R)-12-((1-((4-(difluoromethylidene)piperidin-1-yl)methyl)cyclopropyl)methoxy)-1-fluoro-5-methyl-5a,6,7,8,9,10-hexahydro-5H-4-oxa-3,10a,11,13,14-pentaaza-6,9-methanaphtho[1,8-ab]heptyl-2-yl)-7-fluorobenzo[b]thiophene-3-carbonitrile is reacted with a hydrogen chloride/dioxane solution to form the hydrochloride salt.

In some embodiments, the compound 2-amino-4-((5S,5aS,6S,9R)-12-((1-((4-(difluoromethylidene)piperidin-1-yl)methyl)cyclopropyl)methoxy)-1-fluoro-5-methyl-5a,6,7,8,9,10-hexahydro-5H-4-oxa-3,10a,11,13,14-pentaaza-6,9-methano[1,8-ab]heptyl-2-yl)-7-fluorobenzo[b]thiophene-3-carbonitrile is reacted with a phosphoric acid/ethanol solution to form a phosphate salt.

In some embodiments, the compound 2-amino-4-((5S,5aS,6S,9R)-12-((1-((4-(difluoromethylidene)piperidin-1-yl)methyl)cyclopropyl)methoxy)-1-fluoro-5-methyl-5a,6,7,8,9,10-hexahydro-5H-4-oxa-3,10a,11,13,14-pentaaza-6,9-methanaphtho[1,8-ab]heptyl-2-yl)-7-fluorobenzo[b]thiophene-3-carbonitrile is reacted with an L-tartaric acid/ethanol solution to form the tartrate salt.

In some embodiments, the solvent used in the salt-forming reaction is at least one selected from methanol, ethanol, acetonitrile, ethyl acetate, methyl isobutyl ketone and 2-methyltetrahydrofuran.

On the other hand, the present disclosure also provides a crystalline form A of the hydrochloride salt of the compound 2-amino-4-((5S,5aS,6S,9R)-12-((1-((4-(difluoromethylidene)piperidin-1-yl)methyl)cyclopropyl)methoxy)-1-fluoro-5-methyl-5a,6,7,8,9,10-hexahydro-5H-4-oxa-3,10a,11,13,14-pentaaza-6,9-methanaptho[1,8-ab]heptylcyclo-2-yl)-7-fluorobenzo[b]thiophene-3-carbonitrile, the X-ray powder diffraction pattern expressed in terms of a diffraction angle 2θ having characteristic peaks at 4.765, 9.445, 14.148, 14.671, and 18.874.

In some embodiments, the hydrochloride salt form A has an X-ray powder diffraction pattern represented by a diffraction angle of 2θ, with characteristic peaks at 4.765, 9.058, 9.445, 14.148, 14.671, and 18.874.

In other embodiments, the X-ray powder diffraction pattern of the hydrochloride salt form A expressed in terms of a diffraction angle 2θ is shown in FIG1 .

The present disclosure also provides a method for preparing the hydrochloride salt form A of the aforementioned compound, comprising the steps of: (a) mixing the compound 2-amino-4-((5S,5aS,6S,9R)-12-((1-((4-(difluoromethylidene)piperidin-1-yl)methyl)cyclopropyl)methoxy)-1-fluoro-5-methyl-5a,6,7,8,9,10-hexahydro-5H-4-oxa-3,10a,11,13,14-pentaaza-6,9-methylnaphtho[1,8-ab]heptyl-2-yl)-7-fluorobenzo[b]thiophene-3-carbonitrile and methanol; and (b) adding an HCl/dioxane solution and stirring.

On the other hand, the present disclosure also provides a hydrochloride salt form B of the compound 2-amino-4-((5S,5aS,6S,9R)-12-((1-((4-(difluoromethylidene)piperidin-1-yl)methyl)cyclopropyl)methoxy)-1-fluoro-5-methyl-5a,6,7,8,9,10-hexahydro-5H-4-oxa-3,10a,11,13,14-pentaaza-6,9-methylnaphtho[1,8-ab]heptyl-2-yl)-7-fluorobenzo[b]thiophene-3-carbonitrile, the X-ray powder diffraction pattern expressed in diffraction angle 2θ degrees has characteristic peaks at 5.048, 8.691, 10.031, 14.977, 16.254, and 19.987.

In some embodiments, the hydrochloride salt form B has an X-ray powder diffraction pattern represented by a diffraction angle of 2θ, with characteristic peaks at 5.048, 8.691, 10.031, 12.987, 14.977, 16.254, 19.737, and 19.987.

In some embodiments, the hydrochloride salt form B has an X-ray powder diffraction pattern expressed as a diffraction angle 2θ, with characteristic peaks at 5.048, 8.691, 10.031, 12.987, 14.977, 16.254, 17.138, 19.737, 19.987, and 28.214.

In other embodiments, the X-ray powder diffraction pattern of the hydrochloride salt form B expressed in terms of a diffraction angle of 2θ is shown in FIG2 .

The present disclosure also provides a method for preparing the hydrochloride form B of the aforementioned compound, comprising the steps of: (a) mixing the compound 2-amino-4-((5S,5aS,6S,9R)-12-((1-((4-(difluoromethylidene)piperidin-1-yl)methyl)cyclopropyl)methoxy)-1-fluoro-5-methyl-5a,6,7,8,9,10-hexahydro-5H-4-oxa-3,10a,11,13,14-pentaaza-6,9-methylnaphtho[1,8-ab]heptyl-2-yl)-7-fluorobenzo[b]thiophene-3-carbonitrile and acetonitrile; and (b) adding an HCl/dioxane solution and stirring.

On the other hand, the present disclosure also provides a hydrochloride salt form C of the compound 2-amino-4-((5S,5aS,6S,9R)-12-((1-((4-(difluoromethylidene)piperidin-1-yl)methyl)cyclopropyl)methoxy)-1-fluoro-5-methyl-5a,6,7,8,9,10-hexahydro-5H-4-oxa-3,10a,11,13,14-pentaaza-6,9-methylnaphtho[1,8-ab]heptylcyclo-2-yl)-7-fluorobenzo[b]thiophene-3-carbonitrile, and an X-ray powder diffraction pattern expressed in terms of a diffraction angle 2θ, having characteristic peaks at 17.222, 7.804, and 7.526.

In other embodiments, the X-ray powder diffraction pattern of the hydrochloride salt form C expressed in terms of a diffraction angle 2θ is shown in FIG3 .

The present disclosure also provides a method for preparing the hydrochloride salt form C of the aforementioned compound, comprising the steps of: (a) mixing the compound 2-amino-4-((5S,5aS,6S,9R)-12-((1-((4-(difluoromethylidene)piperidin-1-yl)methyl)cyclopropyl)methoxy)-1-fluoro-5-methyl-5a,6,7,8,9,10-hexahydro-5H-4-oxa-3,10a,11,13,14-pentaaza-6,9-methylnaphtho[1,8-ab]heptyl-2-yl)-7-fluorobenzo[b]thiophene-3-carbonitrile and ethyl acetate; and (b) adding an HCl/dioxane solution and stirring.

On the other hand, the present disclosure also provides a sulfate salt crystalline form A of the compound 2-amino-4-((5S,5aS,6S,9R)-12-((1-((4-(difluoromethylidene)piperidin-1-yl)methyl)cyclopropyl)methoxy)-1-fluoro-5-methyl-5a,6,7,8,9,10-hexahydro-5H-4-oxa-3,10a,11,13,14-pentaaza-6,9-methylnaphtho[1,8-ab]heptylcyclo-2-yl)-7-fluorobenzo[b]thiophene-3-carbonitrile, the X-ray powder diffraction pattern expressed in diffraction angle 2θ degrees having characteristic peaks at 4.722, 9.103, 12.864, 14.146, 14.704, and 18.899.

In some embodiments, the sulfate salt crystal form A has an X-ray powder diffraction pattern represented by a diffraction angle of 2θ, with characteristic peaks at 4.722, 9.103, 12.864, 14.146, 14.704, 17.681, 18.899, and 20.897.

In some embodiments, the sulfate salt crystal form A has an X-ray powder diffraction pattern represented by a diffraction angle of 2θ, with characteristic peaks at 4.722, 9.103, 12.864, 14.146, 14.704, 17.681, 18.899, 20.897, 22.102, and 25.406.

In other embodiments, the X-ray powder diffraction pattern of the sulfate salt crystal form A expressed in terms of a diffraction angle 2θ is shown in FIG4 .

The present disclosure also provides a method for preparing the sulfate salt crystal form A of the aforementioned compound, comprising the steps of (a) mixing the compound 2-amino-4-((5S, 5aS, 6S, 9R)-12-((1-((4-(difluoromethylidene)piperidin-1-yl)methyl)cyclopropyl)methoxy)-1-fluoro-5-methyl-5a,6,7,8,9,10-hexahydro-5H-4-oxa-3,10a,11,13,14-pentaaza-6,9-methylnaphtho[1,8-ab]heptylcyclo-2-yl)-7-fluorobenzo[b]thiophene-3-carbonitrile and acetonitrile, and (b) adding a sulfuric acid/ethanol solution and stirring.

On the other hand, the present disclosure also provides a phosphate crystal form A of the compound 2-amino-4-((5S,5aS,6S,9R)-12-((1-((4-(difluoromethylidene)piperidin-1-yl)methyl)cyclopropyl)methoxy)-1-fluoro-5-methyl-5a,6,7,8,9,10-hexahydro-5H-4-oxa-3,10a,11,13,14-pentaaza-6,9-methylnaphtho[1,8-ab]heptylcyclo-2-yl)-7-fluorobenzo[b]thiophene-3-carbonitrile, the X-ray powder diffraction pattern expressed in diffraction angle 2θ degrees has characteristic peaks at 4.735, 9.012, 14.150, 14.661, and 18.894.

In some embodiments, the phosphate crystal form A has an X-ray powder diffraction pattern represented by a diffraction angle of 2θ, with characteristic peaks at 4.735, 9.012, 11.783, 12.846, 14.150, 14.661, and 18.894.

In some embodiments, the phosphate crystal form A has an X-ray powder diffraction pattern represented by a diffraction angle of 2θ, with characteristic peaks at 4.735, 9.012, 11.783, 12.846, 14.150, 14.661, 18.894, 20.947, and 22.072.

In other embodiments, the X-ray powder diffraction pattern of the phosphate crystal form A expressed in terms of a diffraction angle 2θ is shown in FIG5 .

The present disclosure also provides a method for preparing the phosphate salt A crystal form of the aforementioned compound, comprising the steps of (a) mixing the compound 2-amino-4-((5S,5aS,6S,9R)-12-((1-((4-(difluoromethylidene)piperidin-1-yl)methyl)cyclopropyl)methoxy)-1-fluoro-5-methyl-5a,6,7,8,9,10-hexahydro-5H-4-oxa-3,10a,11,13,14-pentaaza-6,9-methylnaphtho[1,8-ab]heptylcyclo-2-yl)-7-fluorobenzo[b]thiophene-3-carbonitrile and ethanol, and (b) adding phosphoric acid and stirring.

On the other hand, the present disclosure also provides a phosphate form B of the compound 2-amino-4-((5S,5aS,6S,9R)-12-((1-((4-(difluoromethylidene)piperidin-1-yl)methyl)cyclopropyl)methoxy)-1-fluoro-5-methyl-5a,6,7,8,9,10-hexahydro-5H-4-oxa-3,10a,11,13,14-pentaaza-6,9-methanaptho[1,8-ab]heptyl-2-yl)-7-fluorobenzo[b]thiophene-3-carbonitrile, and an X-ray powder diffraction pattern expressed in diffraction angle 2θ degrees, having characteristic peaks at 5.001, 8.678, 11.020, 12.981, 14.846, and 19.704.

In some embodiments, the phosphate B crystal form has an X-ray powder diffraction pattern represented by a diffraction angle of 2θ, with characteristic peaks at 5.001, 8.678, 11.020, 12.981, 14.846, 16.242, 19.704, and 21.356.

In some embodiments, the phosphate B crystal form has an X-ray powder diffraction pattern represented by a diffraction angle of 2θ, with characteristic peaks at 5.001, 8.678, 11.020, 12.981, 14.846, 16.242, 16.723, 19.704, 21.356, and 22.508.

In other embodiments, the X-ray powder diffraction pattern of the phosphate B crystal form expressed in terms of a diffraction angle 2θ is shown in FIG6 .

The present disclosure also provides a method for preparing the phosphate B crystal form of the aforementioned compound, comprising the steps of (a) mixing the compound 2-amino-4-((5S,5aS,6S,9R)-12-((1-((4-(difluoromethylidene)piperidin-1-yl)methyl)cyclopropyl)methoxy)-1-fluoro-5-methyl-5a,6,7,8,9,10-hexahydro-5H-4-oxa-3,10a,11,13,14-pentaaza-6,9-methylnaphtho[1,8-ab]heptylcyclo-2-yl)-7-fluorobenzo[b]thiophene-3-carbonitrile and acetonitrile, and (b) adding phosphoric acid and stirring.

On the other hand, the present disclosure also provides a crystalline form A of the L-tartrate salt of the compound 2-amino-4-((5S,5aS,6S,9R)-12-((1-((4-(difluoromethylidene)piperidin-1-yl)methyl)cyclopropyl)methoxy)-1-fluoro-5-methyl-5a,6,7,8,9,10-hexahydro-5H-4-oxa-3,10a,11,13,14-pentaaza-6,9-methylnaphtho[1,8-ab]heptylcyclo-2-yl)-7-fluorobenzo[b]thiophene-3-carbonitrile, the X-ray powder diffraction pattern expressed in terms of a diffraction angle 2θ having characteristic peaks at 5.687, 7.991, 10.409, and 12.828.

In some embodiments, the L-tartrate salt form A has an X-ray powder diffraction pattern represented by a diffraction angle of 2θ, with characteristic peaks at 5.687, 7.991, 10.409, 12.828, 16.244, 20.083, and 21.427.

In other embodiments, the X-ray powder diffraction pattern of the L-tartrate salt form A expressed in terms of a diffraction angle of 2θ is shown in FIG7 .

The present disclosure also provides a method for preparing the L-tartrate salt form A of the aforementioned compound, comprising the steps of mixing the compound 2-amino-4-((5S, 5aS, 6S, 9R)-12-((1-((4-(difluoromethylidene)piperidin-1-yl)methyl)cyclopropyl)methoxy)-1-fluoro-5-methyl-5a,6,7,8,9,10-hexahydro-5H-4-oxa-3,10a,11,13,14-pentaaza-6,9-methylnaphtho[1,8-ab]heptylcyclo-2-yl)-7-fluorobenzo[b]thiophene-3-carbonitrile and a solvent (1), (b) adding L-tartaric acid, and stirring, wherein the solvent (1) is selected from methanol.

On the other hand, the present disclosure also provides a crystalline form A of the citrate salt of the compound 2-amino-4-((5S,5aS,6S,9R)-12-((1-((4-(difluoromethylidene)piperidin-1-yl)methyl)cyclopropyl)methoxy)-1-fluoro-5-methyl-5a,6,7,8,9,10-hexahydro-5H-4-oxa-3,10a,11,13,14-pentaaza-6,9-methanaphtho[1,8-ab]heptylcyclo-2-yl)-7-fluorobenzo[b]thiophene-3-carbonitrile, the X-ray powder diffraction pattern expressed in terms of a diffraction angle 2θ having characteristic peaks at 7.276, 15.823, 17.940, 18.247, 19.645, and 21.863.

In some embodiments, the citrate salt form A has an X-ray powder diffraction pattern represented by a diffraction angle of 2θ, with characteristic peaks at 7.276, 15.823, 17.940, 18.247, 19.645, 21.863, 30.027, and 35.408.

In some embodiments, the citrate salt form A has an X-ray powder diffraction pattern expressed as a diffraction angle of 2θ, with characteristic peaks at 5.120, 7.276, 15.823, 17.305, 17.940, 18.247, 19.645, 21.863, 30.027, and 35.408.

In other embodiments, the X-ray powder diffraction pattern of the citrate salt form A expressed in terms of a diffraction angle of 2θ is shown in FIG8 .

The present disclosure also provides a method for preparing the citrate salt form A of the aforementioned compound, comprising the steps of (a) mixing the compound 2-amino-4-((5S,5aS,6S,9R)-12-((1-((4-(difluoromethylidene)piperidin-1-yl)methyl)cyclopropyl)methoxy)-1-fluoro-5-methyl-5a,6,7,8,9,10-hexahydro-5H-4-oxa-3,10a,11,13,14-pentaaza-6,9-methylnaphtho[1,8-ab]heptylcyclo-2-yl)-7-fluorobenzo[b]thiophene-3-carbonitrile and ethyl acetate, and (b) adding citric acid and stirring.

On the other hand, the present disclosure also provides a citrate salt form B of the compound 2-amino-4-((5S,5aS,6S,9R)-12-((1-((4-(difluoromethylidene)piperidin-1-yl)methyl)cyclopropyl)methoxy)-1-fluoro-5-methyl-5a,6,7,8,9,10-hexahydro-5H-4-oxa-3,10a,11,13,14-pentaaza-6,9-methanaptho[1,8-ab]heptyl-2-yl)-7-fluorobenzo[b]thiophene-3-carbonitrile, the X-ray powder diffraction pattern expressed in diffraction angle 2θ degrees having characteristic peaks at 8.663, 19.723, 14.863, and 10.973.

In some embodiments, the citrate salt form B has an X-ray powder diffraction pattern represented by a diffraction angle of 2θ, with characteristic peaks at 8.663, 10.973, 14.863, 16.264, 19.723, and 22.586.

In other embodiments, the X-ray powder diffraction pattern of the citrate salt form B expressed in terms of a diffraction angle of 2θ is shown in FIG9 .

The present disclosure also provides a method for preparing the citrate salt form B of the aforementioned compound, comprising the steps of (a) mixing the compound 2-amino-4-((5S,5aS,6S,9R)-12-((1-((4-(difluoromethylidene)piperidin-1-yl)methyl)cyclopropyl)methoxy)-1-fluoro-5-methyl-5a,6,7,8,9,10-hexahydro-5H-4-oxa-3,10a,11,13,14-pentaaza-6,9-methylnaphtho[1,8-ab]heptylcyclo-2-yl)-7-fluorobenzo[b]thiophene-3-carbonitrile and acetonitrile, and (b) adding citric acid and stirring.

On the other hand, the present disclosure also provides a citrate salt form C of the compound 2-amino-4-((5S,5aS,6S,9R)-12-((1-((4-(difluoromethylidene)piperidin-1-yl)methyl)cyclopropyl)methoxy)-1-fluoro-5-methyl-5a,6,7,8,9,10-hexahydro-5H-4-oxa-3,10a,11,13,14-pentaaza-6,9-methanaptho[1,8-ab]heptylcyclo-2-yl)-7-fluorobenzo[b]thiophene-3-carbonitrile, the X-ray powder diffraction pattern expressed in diffraction angle 2θ degrees having characteristic peaks at 7.076, 7.483, 15.665, 17.203, and 19.775.

In some embodiments, the citrate salt form C has an X-ray powder diffraction pattern represented by a diffraction angle of 2θ, with characteristic peaks at 4.952, 7.076, 7.483, 13.147, 15.665, 17.203, and 19.775.

In some embodiments, the citrate salt form C has an X-ray powder diffraction pattern represented by a diffraction angle of 2θ, with characteristic peaks at 4.952, 7.076, 7.483, 13.147, 15.665, 16.499, 17.203, 17.746, and 19.775.

In other embodiments, the X-ray powder diffraction pattern of the citrate salt form C expressed in terms of a diffraction angle of 2θ is shown in FIG10 .

On the other hand, the present disclosure also provides a citrate salt form D of the compound 2-amino-4-((5S,5aS,6S,9R)-12-((1-((4-(difluoromethylidene)piperidin-1-yl)methyl)cyclopropyl)methoxy)-1-fluoro-5-methyl-5a,6,7,8,9,10-hexahydro-5H-4-oxa-3,10a,11,13,14-pentaaza-6,9-methylnaphtho[1,8-ab]heptyl-2-yl)-7-fluorobenzo[b]thiophene-3-carbonitrile, the X-ray powder diffraction pattern expressed in diffraction angle 2θ degrees having characteristic peaks at 4.728, 9.421, 14.12, 14.619, and 18.856.

In some embodiments, the citrate salt form D has an X-ray powder diffraction pattern expressed as a diffraction angle 2θ, with characteristic peaks at 4.728, 9.421, 14.12, 14.619, 18.856, 20.922, 29.106, and 33.272.

In other embodiments, the X-ray powder diffraction pattern of the citrate salt form D expressed in terms of a diffraction angle of 2θ is shown in FIG11 .

The present disclosure also provides a method for preparing the citrate D crystalline form of the aforementioned compound, comprising the steps of (a) mixing the compound 2-amino-4-((5S,5aS,6S,9R)-12-((1-((4-(difluoromethylidene)piperidin-1-yl)methyl)cyclopropyl)methoxy)-1-fluoro-5-methyl-5a,6,7,8,9,10-hexahydro-5H-4-oxa-3,10a,11,13,14-pentaaza-6,9-methylnaphtho[1,8-ab]heptylcyclo-2-yl)-7-fluorobenzo[b]thiophene-3-carbonitrile and ethanol, and (b) adding citric acid and stirring.

On the other hand, the present disclosure also provides a crystalline form A of the methanesulfonate of the compound 2-amino-4-((5S,5aS,6S,9R)-12-((1-((4-(difluoromethylidene)piperidin-1-yl)methyl)cyclopropyl)methoxy)-1-fluoro-5-methyl-5a,6,7,8,9,10-hexahydro-5H-4-oxa-3,10a,11,13,14-pentaaza-6,9-methylnaphtho[1,8-ab]heptylcyclo-2-yl)-7-fluorobenzo[b]thiophene-3-carbonitrile, the X-ray powder diffraction pattern expressed in terms of a diffraction angle 2θ having characteristic peaks at 7.553, 9.757, and 13.097.

In some embodiments, the mesylate salt form A has an X-ray powder diffraction pattern expressed as a diffraction angle 2θ, with characteristic peaks at 7.553, 9.757, 13.097, 16.859, 18.394, 19.738, 20.429, and 21.082.

In other embodiments, the X-ray powder diffraction pattern of the mesylate salt form A expressed in terms of a diffraction angle of 2θ is shown in FIG12 .

The present disclosure also provides a method for preparing a crystalline form A of a mesylate salt of the aforementioned compound, comprising the steps of mixing the compound 2-amino-4-((5S,5aS,6S,9R)-12-((1-((4-(difluoromethylidene)piperidin-1-yl)methyl)cyclopropyl)methoxy)-1-fluoro-5-methyl-5a,6,7,8,9,10-hexahydro-5H-4-oxa-3,10a,11,13,14-pentaaza-6,9-methylnaphtho[1,8-ab]heptylcyclo-2-yl)-7-fluorobenzo[b]thiophene-3-carbonitrile and a solvent (2), (b) adding methanesulfonic acid, and then adding a solvent (3), and stirring, wherein the solvent (2) is selected from 2-methyltetrahydrofuran, and the solvent (3) is selected from n-heptane.

On the other hand, the present disclosure also provides a crystalline form A of the benzoate salt of the compound 2-amino-4-((5S,5aS,6S,9R)-12-((1-((4-(difluoromethylidene)piperidin-1-yl)methyl)cyclopropyl)methoxy)-1-fluoro-5-methyl-5a,6,7,8,9,10-hexahydro-5H-4-oxa-3,10a,11,13,14-pentaaza-6,9-methanaptho[1,8-ab]heptylcyclo-2-yl)-7-fluorobenzo[b]thiophene-3-carbonitrile, the X-ray powder diffraction pattern expressed in terms of a diffraction angle 2θ having characteristic peaks at 7.515, 8.359, and 11.945.

In some embodiments, the benzoate salt form A has an X-ray powder diffraction pattern represented by a diffraction angle of 2θ, with characteristic peaks at 7.515, 8.002, 8.359, 11.945, 12.920, and 18.970.

In other embodiments, the X-ray powder diffraction pattern of the benzoate salt form A expressed in terms of a diffraction angle of 2θ is shown in FIG13 .

The present disclosure also provides a method for preparing a crystalline form A of the benzoate salt of the aforementioned compound, comprising the steps of mixing the compound 2-amino-4-((5S,5aS,6S,9R)-12-((1-((4-(difluoromethylidene)piperidin-1-yl)methyl)cyclopropyl)methoxy)-1-fluoro-5-methyl-5a,6,7,8,9,10-hexahydro-5H-4-oxa-3,10a,11,13,14-pentaaza-6,9-methylnaphtho[1,8-ab]heptylcyclo-2-yl)-7-fluorobenzo[b]thiophene-3-carbonitrile and a solvent (4), (b) adding benzoic acid, and stirring, wherein the solvent (4) is selected from methyl isobutyl ketone or 2-methyltetrahydrofuran.

On the other hand, the present disclosure also provides a crystalline form A of the succinate salt of the compound 2-amino-4-((5S,5aS,6S,9R)-12-((1-((4-(difluoromethylidene)piperidin-1-yl)methyl)cyclopropyl)methoxy)-1-fluoro-5-methyl-5a,6,7,8,9,10-hexahydro-5H-4-oxa-3,10a,11,13,14-pentaaza-6,9-methylnaphtho[1,8-ab]heptyl-2-yl)-7-fluorobenzo[b]thiophene-3-carbonitrile, the X-ray powder diffraction pattern expressed in terms of a diffraction angle 2θ having characteristic peaks at 7.943, 10.351, 12.828, 16.138, 19.832, and 21.057.

In some embodiments, the succinate salt form A has an X-ray powder diffraction pattern represented by a diffraction angle of 2θ, with characteristic peaks at 7.943, 10.351, 12.828, 13.212, 16.138, 19.832, 21.057, and 23.619.

In some embodiments, the succinate salt form A has an X-ray powder diffraction pattern expressed as a diffraction angle 2θ, with characteristic peaks at 6.685, 7.943, 10.351, 12.828, 13.212, 16.138, 19.832, 21.057, 21.465, and 23.619.

In other embodiments, the X-ray powder diffraction pattern of the succinate salt form A expressed in terms of a diffraction angle of 2θ is shown in FIG14 .

The present disclosure also provides a method for preparing a crystalline form A of the succinate salt of the aforementioned compound, comprising the steps of mixing the compound 2-amino-4-((5S, 5aS, 6S, 9R)-12-((1-((4-(difluoromethylidene)piperidin-1-yl)methyl)cyclopropyl)methoxy)-1-fluoro-5-methyl-5a,6,7,8,9,10-hexahydro-5H-4-oxa-3,10a,11,13,14-pentaaza-6,9-methylnaphtho[1,8-ab]heptylcyclo-2-yl)-7-fluorobenzo[b]thiophene-3-carbonitrile and a solvent (5), (b) adding succinic acid, and stirring, wherein the solvent (5) is selected from methyl isobutyl ketone.

On the other hand, the present disclosure also provides a crystalline form B of the succinate salt of the compound 2-amino-4-((5S,5aS,6S,9R)-12-((1-((4-(difluoromethylidene)piperidin-1-yl)methyl)cyclopropyl)methoxy)-1-fluoro-5-methyl-5a,6,7,8,9,10-hexahydro-5H-4-oxa-3,10a,11,13,14-pentaaza-6,9-methylnaphtho[1,8-ab]heptylcyclo-2-yl)-7-fluorobenzo[b]thiophene-3-carbonitrile, the X-ray powder diffraction pattern expressed in terms of a diffraction angle 2θ having characteristic peaks at 7.189, 7.976, 8.802, 10.681, and 15.899.

In some embodiments, the succinate salt form B has an X-ray powder diffraction pattern represented by a diffraction angle of 2θ, with characteristic peaks at 7.189, 7.976, 8.802, 10.681, 15.899, 22.331, and 28.635.

In some embodiments, the succinate salt form B has an X-ray powder diffraction pattern expressed as a diffraction angle of 2θ, with characteristic peaks at 4.420, 4.989, 7.189, 7.976, 8.802, 10.681, 15.899, 22.331, and 28.635.

In other embodiments, the X-ray powder diffraction pattern of the succinate salt form B expressed in terms of a diffraction angle of 2θ is shown in FIG15 .

The present disclosure also provides a method for preparing the succinate salt form B of the aforementioned compound, comprising the steps of (a) mixing the compound 2-amino-4-((5S, 5aS, 6S, 9R)-12-((1-((4-(difluoromethylidene)piperidin-1-yl)methyl)cyclopropyl)methoxy)-1-fluoro-5-methyl-5a,6,7,8,9,10-hexahydro-5H-4-oxa-3,10a,11,13,14-pentaaza-6,9-methylnaphtho[1,8-ab]heptylcyclo-2-yl)-7-fluorobenzo[b]thiophene-3-carbonitrile and acetonitrile, and (b) adding succinic acid and stirring.

On the other hand, the present disclosure also provides a crystalline form C of the succinate salt of the compound 2-amino-4-((5S,5aS,6S,9R)-12-((1-((4-(difluoromethylidene)piperidin-1-yl)methyl)cyclopropyl)methoxy)-1-fluoro-5-methyl-5a,6,7,8,9,10-hexahydro-5H-4-oxa-3,10a,11,13,14-pentaaza-6,9-methylnaphtho[1,8-ab]heptyl-2-yl)-7-fluorobenzo[b]thiophene-3-carbonitrile, which has an X-ray powder diffraction pattern expressed in terms of a diffraction angle 2θ, having characteristic peaks at 5.179, 7.238, 7.525, 13.268, 15.91, and 17.916.

In some embodiments, the succinate salt form C has an X-ray powder diffraction pattern represented by a diffraction angle of 2θ, with characteristic peaks at 5.179, 7.238, 7.525, 13.268, 15.91, 16.681, 17.357, and 17.916.

In some embodiments, the succinate salt form C has an X-ray powder diffraction pattern expressed as a diffraction angle 2θ, with characteristic peaks at 5.179, 7.238, 7.525, 8.693, 10.064, 13.268, 15.91, 16.681, 17.357, 17.916, and 19.45.

In other embodiments, the X-ray powder diffraction pattern of the succinate salt form C expressed in terms of a diffraction angle of 2θ is shown in FIG16 .

The present disclosure also provides a method for preparing the succinate salt crystal form C of the aforementioned compound, comprising the steps of (a) mixing the compound 2-amino-4-((5S, 5aS, 6S, 9R)-12-((1-((4-(difluoromethylidene)piperidin-1-yl)methyl)cyclopropyl)methoxy)-1-fluoro-5-methyl-5a,6,7,8,9,10-hexahydro-5H-4-oxa-3,10a,11,13,14-pentaaza-6,9-methylnaphtho[1,8-ab]heptylcyclo-2-yl)-7-fluorobenzo[b]thiophene-3-carbonitrile and ethyl acetate, and (b) adding succinic acid and stirring.

Furthermore, the present invention discloses an X-ray powder diffraction pattern of the aforementioned crystal form expressed in terms of a diffraction angle 2θ, wherein the error range of the 2θ angle is ±0.2.

On the other hand, the present disclosure also provides a complex comprising the compound 2-amino-4-((5S,5aS,6S,9R)-12-((1-((4-(difluoromethylidene)piperidin-1-yl)methyl)cyclopropyl)methoxy)-1-fluoro-5-methyl-5a,6,7,8,9,10-hexahydro-5H-4-oxa-3,10a,11,13,14-pentaaza-6,9-methylnaphtho[1,8-ab]heptyl-2-yl)-7-fluorobenzo[b]thiophene-3-carbonitrile and saccharin.

In some embodiments, the chemical ratio of the compound 2-amino-4-((5S,5aS,6S,9R)-12-((1-((4-(difluoromethylidene)piperidin-1-yl)methyl)cyclopropyl)methoxy)-1-fluoro-5-methyl-5a,6,7,8,9,10-hexahydro-5H-4-oxa-3,10a,11,13,14-pentaaza-6,9-methanaptho[1,8-ab]heptyl-2-yl)-7-fluorobenzo[b]thiophene-3-carbonitrile to the saccharin molecule is 1:1.

The present disclosure also provides a method for preparing a complex, which includes the step of mixing the compound 2-amino-4-((5S,5aS,6S,9R)-12-((1-((4-(difluoromethylidene)piperidin-1-yl)methyl)cyclopropyl)methoxy)-1-fluoro-5-methyl-5a,6,7,8,9,10-hexahydro-5H-4-oxa-3,10a,11,13,14-pentaaza-6,9-methano[1,8-ab]heptyl-2-yl)-7-fluorobenzo[b]thiophene-3-carbonitrile with saccharin.

In some embodiments, a method for preparing a complex comprises the steps of mixing the compound 2-amino-4-((5S,5aS,6S,9R)-12-((1-((4-(difluoromethylidene)piperidin-1-yl)methyl)cyclopropyl)methoxy)-1-fluoro-5-methyl-5a,6,7,8,9,10-hexahydro-5H-4-oxa-3,10a,11,13,14-pentaaza-6,9-methylnaphtho[1,8-ab]heptyl-2-yl)-7-fluorobenzo[b]thiophene-3-carbonitrile and a solvent (6), (b) adding saccharin, and then adding a solvent (7), and stirring, wherein the solvent (6) is selected from methanol or 2-methyltetrahydrofuran, and the solvent (7) is selected from isopropyl ether or n-heptane.

In certain embodiments, the preparation method disclosed herein further comprises any one of the steps of stirring and dissolving or heating and dissolving, crystallizing, filtering, washing or drying.

In some embodiments, the crystallization includes but is not limited to stirring crystallization, such as dissolution crystallization.

In some embodiments, the drying method includes but is not limited to forced air drying and vacuum drying. The drying temperature is generally 25°C to 100°C, preferably 30°C to 70°C, such as 40°C, 50°C or 60°C.

On the other hand, the present disclosure also provides a pharmaceutical composition comprising the aforementioned pharmaceutically acceptable salt, or the aforementioned crystal form, or a complex and a pharmaceutically acceptable excipient.

The present disclosure also provides a pharmaceutical composition prepared from the aforementioned pharmaceutically acceptable salt, or the aforementioned crystal form, or the complex and a pharmaceutically acceptable excipient.

The present disclosure also provides a method for preparing a pharmaceutical composition, comprising the step of mixing the aforementioned pharmaceutically acceptable salt, or the aforementioned crystal form, or the complex with a pharmaceutically acceptable excipient.

The present disclosure also provides the use of the aforementioned pharmaceutically acceptable salt, or the aforementioned crystal form, or the complex, or the aforementioned pharmaceutical composition in the preparation of a medicament for preventing and/or treating a disease or condition mediated by KRAS G12D. In some embodiments, the disease or condition mediated by KRAS G12D is selected from brain cancer, thyroid cancer, head and neck cancer, nasopharyngeal cancer, pharyngeal cancer, oral cancer, salivary gland cancer, esophageal cancer, gastric cancer, lung cancer, liver cancer, kidney cancer, pancreatic cancer, gallbladder cancer, bile duct cancer, colorectal cancer, small intestine cancer, gastrointestinal stromal tumor, urothelial cancer, urethral cancer, bladder cancer, breast cancer, vaginal cancer, ovarian cancer, endometrial cancer, cervical cancer, fallopian tube cancer, testicular cancer, prostate cancer, hemangioma, leukemia, lymphoma, myeloma, skin cancer, lipoma, bone cancer, soft tissue sarcoma, neurofibroma, glioma, neuroblastoma and glioblastoma.

The present disclosure also provides the use of the aforementioned pharmaceutically acceptable salt, or the aforementioned crystal form, or the complex, or the aforementioned pharmaceutical composition in the preparation of a medicament for preventing and/or treating a tumor. In some embodiments, the tumor is selected from brain cancer, thyroid cancer, head and neck cancer, nasopharyngeal cancer, pharyngeal cancer, oral cancer, salivary gland cancer, esophageal cancer, gastric cancer, lung cancer, liver cancer, kidney cancer, pancreatic cancer, gallbladder cancer, bile duct cancer, colorectal cancer, small intestine cancer, gastrointestinal stromal tumor, urothelial cancer, urethral cancer, bladder cancer, breast cancer, vaginal cancer, ovarian cancer, endometrial cancer, cervical cancer, fallopian tube cancer, testicular cancer, prostate cancer, hemangioma, leukemia, lymphoma, myeloma, skin cancer, lipoma, bone cancer, soft tissue sarcoma, neurofibroma, glioma, neuroblastoma and glioblastoma. In other embodiments, the tumor is selected from pancreatic cancer, colorectal cancer and non-small cell lung cancer.

The present disclosure also provides a method for preventing and/or treating a disease or condition mediated by KRAS G12D, which comprises administering the aforementioned pharmaceutically acceptable salt, or the aforementioned crystal form, or the complex, or the aforementioned pharmaceutical composition to a patient.

The present disclosure also provides a method for preventing and/or treating tumors, which comprises administering to a patient the aforementioned pharmaceutically acceptable salt, or the aforementioned crystal form, or the complex, or the aforementioned pharmaceutical composition. In some embodiments, the tumor is selected from brain cancer, thyroid cancer, head and neck cancer, nasopharyngeal cancer, pharyngeal cancer, oral cancer, salivary gland cancer, esophageal cancer, gastric cancer, lung cancer, liver cancer, kidney cancer, pancreatic cancer, gallbladder cancer, bile duct cancer, colorectal cancer, small intestine cancer, gastrointestinal stromal tumor, urothelial cancer, urethral cancer, bladder cancer, breast cancer, vaginal cancer, ovarian cancer, endometrial cancer, cervical cancer, fallopian tube cancer, testicular cancer, prostate cancer, hemangioma, leukemia, lymphoma, myeloma, skin cancer, lipoma, bone cancer, soft tissue sarcoma, neurofibroma, glioma, neuroblastoma and glioblastoma. In other embodiments, the tumor is selected from pancreatic cancer, colorectal cancer and non-small cell lung cancer.

On the other hand, the present disclosure also provides the aforementioned pharmaceutically acceptable salt, or the aforementioned crystal form, or complex, or the aforementioned pharmaceutical composition for preventing and/or treating diseases or conditions mediated by KRAS G12D.

On the other hand, the present disclosure also provides the aforementioned pharmaceutically acceptable salt, or the aforementioned crystal form, or the complex, or the aforementioned pharmaceutical composition for preventing and/or treating tumors. In some embodiments, the tumor is selected from brain cancer, thyroid cancer, head and neck cancer, nasopharyngeal cancer, pharyngeal cancer, oral cancer, salivary gland cancer, esophageal cancer, gastric cancer, lung cancer, liver cancer, kidney cancer, pancreatic cancer, gallbladder cancer, bile duct cancer, colorectal cancer, small intestine cancer, gastrointestinal stromal tumor, urothelial cancer, urethral cancer, bladder cancer, breast cancer, vaginal cancer, ovarian cancer, endometrial cancer, cervical cancer, fallopian tube cancer, testicular cancer, prostate cancer, hemangioma, leukemia, lymphoma, myeloma, skin cancer, lipoma, bone cancer, soft tissue sarcoma, neurofibroma, glioma, neuroblastoma and glioblastoma. In other embodiments, the tumor is selected from pancreatic cancer, colorectal cancer and non-small cell lung cancer.

The "2θ or 2θ angle" mentioned in the present disclosure refers to the diffraction angle, θ is the Bragg angle, and the unit is ° or degree; the error range of each characteristic peak 2θ is ±0.20 (including the case where the number exceeding 1 decimal place is rounded off), specifically -0.20, -0.19, -0.18, -0.17, -0.16, -0.15, -0.14, -0.13, -0.12, -0.11, -0.10, -0.09, -0.08, -0.07, -0.06, -0.05, -0.04, -0.03, -0.02, -0.01, 0.00, 0.01, 0.02, 0.03, 0.04, 0.05, 0.06, 0.07, 0.08, 0.09, 0.10, 0.11, 0.12, 0.13, 0.14, 0.15, 0.16, 0.17, 0.18, 0.19, 0.20.

The numerical values in this disclosure are instrumental measurements or calculated values after instrumental measurement, and are subject to a certain degree of error. Generally speaking, a value within a reasonable error range of plus or minus 10% is within the reasonable error range. Of course, the context in which the numerical value is used must be considered. For example, the total impurity content, which is a value with an error variation of no more than plus or minus 10% after measurement, can be plus or minus 9%, plus or minus 8%, plus or minus 7%, plus or minus 6%, plus or minus 5%, plus or minus 4%, plus or minus 3%, plus or minus 2%, or plus or minus 1%, preferably plus or minus 5%.

The "differential scanning calorimetry or DSC" described in this disclosure refers to measuring the temperature difference and heat flow difference between a sample and a reference object during the process of heating or maintaining the sample at a constant temperature to characterize all physical and chemical changes related to thermal effects and obtain phase change information of the sample.

The drying temperature in the present disclosure is generally 25°C-100°C, preferably 30°C-70°C, and can be dried under normal pressure or reduced pressure.

A "complex" refers to a substance in which two compounds of different types are connected by non-covalent bonds. For example, the two compounds are connected by at least one of hydrogen bonds, van der Waals forces, or π-π forces.

The "pharmaceutically acceptable excipients" described in this disclosure include, but are not limited to, any adjuvant, carrier, glidant, sweetener, diluent, preservative, dye/colorant, flavoring agent, surfactant, wetting agent, dispersant, suspending agent, stabilizer, isotonic agent or emulsifier approved by the U.S. Food and Drug Administration for use by humans or livestock animals.

BRIEF DESCRIPTION OF THE DRAWINGS

Figure 1 is the XRPD spectrum of Form A of the hydrochloride salt of Compound A.

Figure 2 is the XRPD spectrum of the hydrochloride form B of compound A.

Figure 3 is the XRPD spectrum of Form C of the hydrochloride salt of Compound A.

Figure 4 is the XRPD spectrum of the sulfate salt form A of compound A.

Figure 5 is the XRPD spectrum of the phosphate A crystal form of compound A.

Figure 6 is an XRPD spectrum of the phosphate B crystal form of compound A.

Figure 7 is the XRPD spectrum of L-tartrate salt Form A of Compound A.

Figure 8 is an XRPD spectrum of Form A of the citrate salt of Compound A.

Figure 9 is an XRPD spectrum of the citrate salt Form B of Compound A.

Figure 10 is an XRPD spectrum of Form C of the citrate salt of Compound A.

Figure 11 is the XRPD spectrum of the citrate salt Form D of Compound A.

FIG12 is an XRPD spectrum of Form A of the mesylate salt of Compound A.

Figure 13 is an XRPD spectrum of Form A of the benzoate salt of Compound A.

Figure 14 is an XRPD spectrum of the succinate salt Form A of Compound A.

Figure 15 is an XRPD spectrum of the succinate salt Form B of Compound A.

Figure 16 is an XRPD spectrum of Form C of the succinate salt of Compound A.

DETAILED DESCRIPTION

The present disclosure is further described in detail by the following examples and experimental examples. These examples and experimental examples are for illustrative purposes only and are not intended to limit the scope of the present disclosure.

Test conditions of the instruments used in the experiment:

The structures of the compounds were confirmed by nuclear magnetic resonance (NMR) and/or mass spectrometry (MS). NMR shifts (δ) are given in units of ^10-6 (ppm). NMR measurements were performed using a Bruker AVANCE-400 NMR spectrometer. The solvents used were deuterated dimethyl sulfoxide (DMSO- _d6 ), deuterated chloroform ( _CDCl3 ), and deuterated methanol ( _CD3OD ), with tetramethylsilane (TMS) as the internal standard.

MS measurements were performed using an Agilent 1200/1290 DAD-6110/6120 Quadrupole MS LC/MS instrument (manufacturer: Agilent, MS model: 6110/6120 Quadrupole MS), a Waters ACQuity UPLC-QD/SQD (manufacturer: Waters, MS model: Waters ACQuity Qda Detector/Waters SQ Detector), and a THERMO Ultimate 3000-Q Exactive (manufacturer: THERMO, MS model: THERMO Q 15 Exactive).

HPLC determinations were performed using an Agilent 1260DAD high pressure liquid chromatograph (Sunfire C18 150×4.6 mm column) and a Thermo U3000 high pressure liquid chromatograph (Gimini C18 150×4.6 mm column).

XRPD is X-ray powder diffraction detection: the measurement is carried out using a BRUKER D8 X-ray diffractometer, specific collection information: Cu anode (40kV, 40mA), radiation: monochromatic Cu-Ka radiation Scanning mode: θ/2θ, scanning range: 3-48°.

DSC stands for differential scanning calorimetry: the measurement was performed using a METTLER TOLEDO DSC 3+ differential scanning calorimeter with a heating rate of 10°C/min, 25-300°C or 25-350°C, and a nitrogen purge rate of 50 mL/min.

TGA stands for thermogravimetric analysis: the test was performed using a METTLER TOLEDO TGA 2 thermogravimetric analyzer with a heating rate of 10°C/min. The specific temperature range was referred to the corresponding spectrum, and the nitrogen purge rate was 50 mL/min.

DVS stands for dynamic moisture adsorption: using Surface Measurement Systems instrinsic, humidity starts from 50%, the humidity range is 0%-95%, the step is 10%, the judgment standard is each gradient mass change dM/dT ≤ 0.002%, TMAX 360min, two cycles.

The known starting materials disclosed herein can be synthesized by methods known in the art, or can be purchased from ABCR GmbH & Co. KG, Acros Organics, Aldrich Chemical Company, Accela ChemBio Inc, Darui Chemicals, etc.

The reaction progress in the examples was monitored by thin layer chromatography (TLC). The developing solvent used in the reaction, the eluent system for column chromatography used to purify the compound, and the developing solvent system for thin layer chromatography included: A: dichloromethane/methanol system, B: n-hexane/ethyl acetate system. The volume ratio of the solvent was adjusted according to the polarity of the compound, and a small amount of alkaline or acidic reagents such as triethylamine and acetic acid could also be added for adjustment.

Example 1

first step

2,5,7-Trichloro-8-fluoropyrido[4,3-d]pyrimidin-4-ol 1b

The crude compound 1a (2 g, 8 mmol) was dissolved in phosphorus oxychloride (25 mL), and N,N-diisopropylethylamine (5.16 g, 40 mmol) was added. The reaction was stirred at 110°C for 14 hours. The reaction solution was cooled to room temperature and concentrated under reduced pressure. The residue was dissolved in 1,4-dioxane, and 20% potassium carbonate solution was added dropwise to adjust the pH to 2-3. The mixture was stirred for 2 hours and then filtered. The filter cake was washed with water and dried to obtain the crude title compound 1b (1.5 g). The product was used directly in the next step without purification.

MS m/z(ESI):267.8[M+1].

Step 2

tert-Butyl (1S,2S,5R)-2-((S)-1-((2,7-dichloro-8-fluoro-4-hydroxypyrido[4,3-d]pyrimidin-5-yl)oxy)ethyl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate 1d

Tert-butyl (1S,2S,5R)-2-((S)-1-hydroxyethyl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate 1c (370 mg, 1.44 mmol, prepared by the method disclosed in Intermediate 29 on page 164 of the specification of patent application "WO2022173678A1")) was dissolved in tetrahydrofuran (10 mL), and sodium hydride (201 mg, 5.2 mmol, 60% purity) was added under ice bath. After reacting for 30 minutes, compound 1b (353 mg, 1.31 mmol) was added and the reaction was stirred for 2 hours. Water was added to the reaction solution to quench the reaction and the solution was concentrated under reduced pressure to obtain the crude title compound 1d (600 mg). The product was used directly in the next step without purification.

MS m/z(ESI):488.2[M+1].

Step 3

(5S,5aS,6S,9R)-2,12-Dichloro-1-fluoro-5-methyl-5a,6,7,8,9,10-hexahydro-5H-4-oxa-3,10a,11,13,14-pentaaza-6,9-methanaphtho[1,8-ab]heptyl-14-carboxylic acid tert-butyl ester 1e

Compound 1d (78 mg, 159.7 μmol) was dissolved in dichloromethane (2 mL). N,N-diisopropylethylamine (61.9 mg, 478.9 μmol) and phosphorus oxychloride (122.4 mg, 798.2 μmol) were added under ice-cooling and stirred for 2 hours. Saturated sodium bicarbonate solution was added to the reaction solution to quench the reaction. The organic phases were combined with dichloromethane (10 mL × 2), dried over anhydrous sodium sulfate, filtered to remove the desiccant, and concentrated under reduced pressure to obtain the crude title compound 1e (75 mg). The product was used directly in the next step without purification.

MS m/z(ESI):470.2[M+1].

Step 4

tert-Butyl (5S,5aS,6S,9R)-12-((1-((tert-butyldimethylsilyl)oxy)methyl)cyclopropyl)methoxy)-2-chloro-1-fluoro-5-methyl-5a,6,7,8,9,10-hexahydro-5H-4-oxa-3,10a,11,13,14-pentaaza-6,9-methanaphtho[1,8-ab]heptyl-14-carboxylate 1 g

(1-(((tert-Butyldimethylsilyl)oxy)methyl)cyclopropyl)methanol 1f (1.4 g, 6.4 mmol) was dissolved in tetrahydrofuran (15 mL), and a 2M solution of sodium bis(trimethylsilyl)amide in tetrahydrofuran was added under ice-cooling. The mixture was stirred at the maintained temperature for 30 minutes. Then, a solution of crude compound 1e (2.3 g, 4.9 mmol) in tetrahydrofuran (20 mL) was added under ice-cooling. The mixture was stirred at the maintained temperature for 1 hour. Saturated ammonium chloride solution was added to the reaction solution to quench the reaction. The mixture was extracted with ethyl acetate (30 mL×2). The organic phases were combined, dried over anhydrous sodium sulfate, filtered to remove the desiccant, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography with eluent System B to obtain the title compound 1g (2 g, yield: 62.8%).

MS m/z(ESI):650.2[M+1].

Step 5

tert-Butyl (5S,5aS,6S,9R)-2-chloro-1-fluoro-12-((1-(hydroxymethyl)cyclopropyl)methoxy)-5-methyl-5a,6,7,8,9,10-hexahydro-5H-4-oxa-3,10a,11,13,14-pentaaza-6,9-methanaphtho[1,8-ab]heptyl-4-carboxylate

Compound 1g (100 mg, 153.8 μmol) was dissolved in tetrahydrofuran (4 mL), and a 1 M tetrabutylammonium fluoride solution in tetrahydrofuran (187 μL) was added. The mixture was stirred for 2 hours. Saturated aqueous ammonium chloride was added to the reaction solution for quenching, and the mixture was extracted with ethyl acetate (15 mL × 3). The organic phases were combined, washed with water and saturated sodium chloride solution, dried over anhydrous sodium sulfate, and the desiccant was removed by filtration. The filtrate was concentrated under reduced pressure to obtain the crude title compound 1h (82 mg), which was used directly in the next step without purification.

MS m/z(ESI):536.2[M+1].

Step 6

tert-Butyl (5S,5aS,6S,9R)-2-chloro-1-fluoro-5-methyl-12-((1-((methylsulfonyl)oxy)methyl)cyclopropyl)methoxy)-5a,6,7,8,9,10-hexahydro-5H-4-oxa-3,10a,11,13,14-pentaaza-6,9-methanaphtho[1,8-ab]heptyl-14-carboxylate 1i

The crude compound 1h (83 mg, 154.9 μmol) and N,N-diisopropylethylamine (60 mg, 464.2 μmol) were dissolved in dichloromethane (3 mL). Methanesulfonyl chloride (25 mg, 218.2 μmol) was added under ice bath and the reaction was allowed to return to room temperature for 30 minutes. Saturated aqueous ammonium chloride was added to the reaction solution for quenching. The mixture was extracted with ethyl acetate (10 mL × 3). The organic phases were combined, washed with water and saturated sodium chloride solution, dried over anhydrous sodium sulfate, and the desiccant was removed by filtration. The filtrate was concentrated under reduced pressure to give the crude title compound 1i (95 mg), which was used directly in the next step without purification.

MS m/z(ESI):614.2[M+1].

Step 7

(5S,5aS,6S,9R)-tert-Butyl 2-chloro-12-((1-((4-(difluoromethylidene)piperidin-1-yl)methyl)cyclopropyl)methoxy)-1-fluoro-5-methyl-5a,6,7,8,9,10-hexahydro-5H-4-oxa-3,10a,11,13,14-pentaaza-6,9-methanaphtho[1,8-ab]heptyl-14-carboxylate 1j

The crude compound 1i (30 mg, 48.8 μmol) and 4-(difluoromethylidene)piperidine hydrochloride (12.4 mg, 73.2 μmol) were dissolved in acetonitrile (4 mL), and anhydrous potassium carbonate (20.2 mg, 146.5 μmol) and sodium iodide (22 mg, 146.5 μmol) were added. The mixture was stirred at 80°C for 1 hour. The reaction solution was cooled to room temperature and filtered. The filtrate was diluted with water and extracted with ethyl acetate (5 mL×3). The organic phases were combined, washed with water and saturated sodium chloride solution in sequence, and dried over anhydrous sodium sulfate. The desiccant was removed by filtration, and the filtrate was concentrated under reduced pressure to obtain the crude title compound 1j (31 mg). The product was used directly in the next step without purification.

MS m/z(ESI):651.2[M+1].

Step 8

(5S,5aS,6S,9R)-tert-Butyl 2-(2-((tert-butoxycarbonyl)amino)-3-cyano-7-fluorobenzo[b]thiophen-4-yl)-12-(1-((4-(difluoromethylidene)piperidin-1-yl)methyl)cyclopropyl)methoxy)-1-fluoro-5-methyl-5a,6,7,8,9,10-hexahydro-5H-4-oxa-3,10a,11,13,14-pentaaza-6,9-methanaphtho[1,8-ab]heptane-14-carboxylate 1k

Compound 1j (31 mg, 47.6 μmol), tert-butyl (3-cyano-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-7-fluorobenzo[b]thiophen-2-yl)carbamate (26.9 mg, 66.6 μmol), tetrakis(triphenylphosphine)palladium (11 mg, 9.5 μmol), and cesium carbonate (46.5 mg, 142.8 μmol) were mixed in N,N-dimethylformamide (1 mL) and reacted at 100°C under a nitrogen atmosphere for 3 hours. The reaction solution was cooled to room temperature and filtered, and the filtrate was concentrated under reduced pressure to give the crude title compound 1k (43 mg), which was used directly in the next step without purification.

MS m/z(ESI):907.2[M+1].

Step 9

2-Amino-4-((5S,5aS,6S,9R)-12-((1-((4-(difluoromethylidene)piperidin-1-yl)methyl)cyclopropyl)methoxy)-1-fluoro-5-methyl-5a,6,7,8,9,10-hexahydro-5H-4-oxa-3,10a,11,13,14-pentaaza-6,9-methano[1,8-ab]heptyl-2-yl)-7-fluorobenzo[b]thiophene-3-carbonitrile (Compound A)

The crude compound 1k (40 mg, 44.1 μmol) was dissolved in dichloromethane (0.5 mL), and trifluoroacetic acid (0.5 mL) was added. After stirring for 1 hour, the reaction was concentrated under reduced pressure. The residue was purified by high performance liquid chromatography (Waters-2545, column: YMC Triart-Exrs C18, 30*150 mm, 5 μm; mobile phase: aqueous phase (10 mmol/L ammonium bicarbonate) and acetonitrile, gradient ratio: acetonitrile 30%-45%, flow rate: 30 mL/min) to give the title compound A (2 mg, yield: 6.4%).

MS m/z(ESI):707.2[M+1].

¹ H NMR (500MHz, CD ₃ OD): δ7.39 (dd, 1H), 7.04 (t, 1H), 5.41 (dd, 1H), 4.91 (s, 3H), 4.54 (d, 1H), 4.48 (d, 1H), 4.39 (d, 1H), 4.10 (d, 1H), 3.71 (d, 1H), 3.61 (s, 1H), 3.19(d, 1H), 2.70-2.41(m, 5H), 2.22(s, 3H), 2.10(q, 2H), 2.01-1.83(m, 2H), 1.79(d, 2H), 1.61(d, 3H), 0.75(s, 2H), 0.52(s, 2H).

Test Example 1: Biological Evaluation of GP2d and AGS Cell 3D Proliferation Inhibition Experiment

1. Test Purpose

The inhibitory effect of the disclosed compounds on the KRAS target was evaluated by testing the 3D proliferation inhibitory effect of the disclosed compounds on GP2d and AGS cells.

2. Experimental Methods

GP2d cells were cultured in complete medium (DMEM/high glucose medium (Hyclone, SH30243.01) supplemented with 10% fetal bovine serum (Corning, 35-076-CV). On the first day of the experiment, GP2d cells were seeded at a density of 1000 cells/well in a 96-well low attachment plate (Corning, CLS7007-24EA) using complete medium. 90 μL of cell suspension was added to each well, centrifuged at 2000 rpm for 5 minutes at room temperature, and then incubated overnight at 37°C in a 5% _CO2 incubator.

AGS cells were cultured in complete medium (RPMI1640 medium (Hyclone, SH30809.01) supplemented with 10% fetal bovine serum (Corning, 35-076-CV). On the first day of the experiment, AGS cells were seeded at a density of 1000 cells/well in a 96-well low attachment plate (Corning, CLS7007-24EA) using complete medium. 90 μL of cell suspension was added to each well. After centrifugation at 2000 rpm for 5 minutes at room temperature, the cells were incubated overnight at 37°C in a 5% _CO2 incubator.

On the second day, 10 μL of the compound to be tested prepared in complete culture medium was added to each well. The final concentration of the compound for GP2d cells was 9 concentration points of 5-fold serial dilution starting from 1 μM, and the final concentration of the compound for AGS cells was 9 concentration points of 5-fold serial dilution starting from 10 μM. A blank control containing 0.5% DMSO was set up. The well plate was placed in a cell culture incubator at 37°C and 5% _CO2 for 5 days. On the seventh day, 50 μL of the compound to be tested was added to each well. 3D Cell Viability Assay reagent (Promega, G9682) was shaken at room temperature in the dark for 25 minutes, then pipetted to mix thoroughly and 100 μL was transferred from each well to a white opaque 96-well plate (PerkinElmer, 6005290). The luminescence signal was read using a multi-function microplate reader (PerkinElmer, EnVision2105).

3. Data Analysis

The _IC50 values of the inhibitory activity of the compounds were calculated using Graphpad Prism software. The _IC50 value of the inhibitory activity of compound AGP2d on 3D cell proliferation was 0.4 nM.

Test Example 2: Biological Evaluation of AsPC-1 Cell 3D Proliferation Inhibition Experiment

On the first day of the experiment, well-grown AsPC-1 cells reaching 70%-80% confluence were digested and resuspended in RPMI 1640 (Hyclone, SH30809.01) supplemented with 10% FBS. The cell density was adjusted to the desired level. 90 μL of the cell suspension was added to each well of a U-shaped low-adhesion 96-well plate (Corning, CLS7007-24EA) for a cell density of 1500 cells/well. The plate was centrifuged at 2500 rpm for 5 minutes and incubated overnight in a 37°C, 5% _CO2 incubator. On the second day, a 20 mM DMSO-dissolved test compound was diluted with DMSO to a starting concentration of 2 mM. This was then serially diluted 5-fold to a total of nine concentration points, with DMSO as a control. The serially diluted compound was then further diluted 20-fold with culture medium. 10 μL of the test compound diluted with culture medium was added to each well of the plate for a final concentration of 10 μM starting at the 10 μM concentration and then serially diluted 5-fold to a total of nine concentration points. The wells containing 0.5% DMSO were set as vehicle control wells, and the wells containing only culture medium and 0.5% DMSO were set as blank control wells. Each concentration of compound and control wells were set up in duplicate, and the final DMSO concentration in each well was 0.5%. After centrifugation at 2500 rpm for 3 minutes, the cell plate was placed in a 37°C, 5% _CO2 incubator for 5 days. On the seventh day, the 96-well cell culture plate was removed and 50 μL of the luminescent cell activity detection reagent CellTiter- 3D Cell Viability Assay (Promega, G9683) was shaken at room temperature in the dark for 25 minutes. After mixing by pipetting up and down, 100 μL was transferred to each well of a white opaque OptiPlate ^™ -96-well plate (PerkinElmer, 6005290). Luminescence signals were read using a multi-function microplate reader (PerkinElmer, EnVision 2105).

Inhibition rate was calculated using the following formula: Inhibition rate = (luminescence value _{vehicle control well} - luminescence value _{test compound} ) / (luminescence value _{vehicle control well} - luminescence value _{blank control well} ) × 100%. GraphPad Prism software was used to plot inhibition rate curves based on compound concentrations and calculate compound _IC50 values. Compound A had an _IC50 of 3.7 nM.

Example 2: Preparation of Compound A Hydrochloride

About 8 mg of compound A was weighed and dissolved in 0.16 mL of methanol. An ethanolic solution of HCl (1.2 mol/L, 9.90 μL) was added. The temperature was raised and lowered in a cycle from 5°C to 40°C (rate ±0.6°C/min). 1.0 mL of isopropyl ether was added and stirred to precipitate. After centrifugation, the solid was vacuum dried to obtain a solid.

X-ray powder diffraction analysis revealed no obvious characteristic peaks in the XRPD spectrum, and ion chromatography analysis revealed a chloride ion content of 8.71%.

Example 3: Preparation of Compound A Hydrochloride Form A

Compound A (50 mg, 70.75 μmol) was weighed and dissolved in 1 mL of ethanol. HCl/dioxane solution (4 M, 18.57 μL) was added and stirred at room temperature for 18 h. The mixture was filtered and dried in vacuo to obtain a solid (25 mg).

X-ray powder diffraction analysis revealed that the XRPD spectrum is shown in FIG1 , and the positions of the characteristic peaks are shown in Table 1 , which is defined as hydrochloride crystal form A.

Table 1

Example 4: Preparation of Compound A Hydrochloride Form B

Compound A (50 mg, 70.75 μmol) was weighed and dissolved in 1 mL of acetonitrile. HCl/dioxane solution (4 M, 18.57 μL) was added and stirred at room temperature for 18 h. The mixture was filtered and dried in vacuo to obtain a solid (26 mg).

The XRPD spectrum of the product is shown in FIG2 , and the positions of its characteristic peaks are shown in Table 2 , which is defined as hydrochloride crystal form B.

Table 2

Example 5: Preparation of Compound A Hydrochloride Form C

Compound A (50 mg, 70.75 μmol) was weighed and dissolved in 1 mL of ethyl acetate. HCl/dioxane solution (4 M, 37 μL) was added and stirred at room temperature for 18 h. The mixture was filtered and dried in vacuo to obtain a solid (24 mg).

The XRPD spectrum is shown in Figure 3 and the characteristic peak positions are shown in Table 3, which is defined as hydrochloride C crystal form.

Table 3

Example 6: Preparation of Compound A Sulfate

About 8 mg of compound A was weighed and dissolved in 0.08 mL of methyl isobutyl ketone. An ethanol solution of sulfuric acid (1.84 mol/L, 6.46 μL) was added. The temperature was raised and lowered in a cycle from 5°C to 40°C (rate ±0.6°C/min). The mixture was stirred to precipitate. After centrifugation, the solid was vacuum dried to obtain a solid.

X-ray powder diffraction analysis revealed no obvious characteristic peaks in the XRPD spectrum, and ion chromatography analysis revealed a sulfate ion content of 19.77%.

Example 7: Preparation of Compound A Sulfate

Approximately 8 mg of compound A was weighed and dissolved in 0.08 mL of methanol. A 1.84 mol/L, 6.46 μL, ethanolic sulfuric acid solution was added. The temperature was cycled from 5°C to 40°C (at a rate of ±0.6°C/min). Stirring allowed precipitation to proceed. After centrifugation, the solid was vacuum-dried to obtain the desired product. X-ray powder diffraction analysis revealed no distinct characteristic peaks in the XRPD spectrum.

Example 8: Preparation of Compound A Sulfate

Approximately 8 mg of compound A was weighed and dissolved in 0.08 mL of 2-methyltetrahydrofuran. An ethanolic solution of sulfuric acid (1.84 mol/L, 6.46 μL) was added. The temperature was cycled from 5°C to 40°C (at a rate of ±0.6°C/min). Stirring allowed precipitation to proceed. After centrifugation, the solid was vacuum-dried to obtain the desired product. X-ray powder diffraction analysis revealed no distinct characteristic peaks in the XRPD spectrum.

Example 9: Preparation of Compound A Sulfate Crystal Form A

Compound A (50 mg, 70.75 μmol) was weighed and dissolved in 1 mL of acetonitrile. A solution of sulfuric acid (7.3 mg, 74.4 μmol) in acetonitrile (dissolved in 0.2 mL of acetonitrile) was added. The mixture was stirred at room temperature for 18 h. The solid was collected by filtration and dried in vacuo to obtain a solid (43 mg).

The XRPD spectrum of the product is shown in FIG4 , and the positions of the characteristic peaks are shown in Table 4 , which is defined as sulfate crystal form A.

Table 4

Example 10: Preparation of Compound A Phosphate

About 8 mg of compound A was weighed and dissolved in 0.16 mL of methanol. An ethanolic solution of phosphoric acid (1.46 mol/L, 8.14 μL) was added. The temperature was raised and lowered in a cycle from 5°C to 40°C (rate ±0.6°C/min). The mixture was stirred to precipitate. After centrifugation, the solid was vacuum dried to obtain a solid.

X-ray powder diffraction analysis revealed no obvious characteristic peaks in the XRPD spectrum, and ion chromatography analysis revealed a phosphate ion content of 23.33%.

Example 11: Preparation of Compound A Phosphate

Approximately 8 mg of compound A was weighed and dissolved in 0.16 mL of methyl isobutyl ketone. An ethanolic solution of phosphoric acid (1.46 mol/L, 8.14 μL) was added. The temperature was cycled from 5°C to 40°C (at a rate of ±0.6°C/min). Stirring allowed precipitation to proceed. After centrifugation, the solid was vacuum-dried to obtain the title product. X-ray powder diffraction analysis revealed no distinct characteristic peaks in the XRPD spectrum.

Example 12: Preparation of Compound A Phosphate

Approximately 8 mg of compound A was weighed and dissolved in 0.16 mL of 2-methyltetrahydrofuran. An ethanolic solution of phosphoric acid (1.46 mol/L, 8.14 μL) was added. The temperature was cycled from 5°C to 40°C (at a rate of ±0.6°C/min). Stirring was performed to precipitate the product. After centrifugation, the solid was vacuum-dried to obtain the desired product. X-ray powder diffraction analysis revealed no distinct characteristic peaks in the XRPD spectrum.

Example 13: Preparation of Compound A Phosphate Form A

Compound A (50 mg, 70.75 μmol) was weighed and dissolved in 1 mL of ethanol. Phosphoric acid (8.6 mg, 74.6 μmol) was added and stirred at 50°C for 2 h. The mixture was cooled to room temperature and stirred for 18 h. The solid was collected by filtration and dried under vacuum to obtain a solid (40 mg).

X-ray powder diffraction analysis revealed that the XRPD spectrum was shown in FIG5 , and the positions of the characteristic peaks were shown in Table 5 , which was defined as phosphate crystal form A.

Table 5

Example 14: Preparation of Compound A Phosphate Form B

Compound A (50 mg, 70.75 μmol) was weighed and dissolved in 1.3 mL of acetonitrile. Phosphoric acid (8.15 mg, 70.7 μmol) was added and stirred at 50°C for 2 h. The mixture was cooled to room temperature and stirred for 18 h. The mixture was filtered and dried in vacuo to obtain a solid (35 mg).

X-ray powder diffraction analysis revealed that the XRPD spectrum was shown in FIG6 , and the positions of the characteristic peaks were shown in Table 6 , which was defined as phosphate B crystal form.

Table 6

Example 15: Preparation of Compound AL-Tartrate Crystal Form A

About 8 mg of compound A was weighed and dissolved in 0.16 mL of methanol. An ethanol solution of L-tartaric acid (1 mol/L, 11.89 μL) was added. The temperature was raised and lowered in a cycle from 5°C to 40°C (rate ±0.6°C/min). The mixture was stirred for crystallization. After centrifugation, the solid was vacuum dried to obtain a solid.

X-ray powder diffraction analysis showed that the product was L-tartrate salt form A. The XRPD spectrum is shown in FIG7 , and the positions of the characteristic peaks are shown in Table 7.

The DSC spectrum showed endothermic peaks at 76.00°C, 128.84°C, and 190.18°C. The TGA spectrum showed a weight loss of 3.47% from 32°C to 134°C and a weight loss of 1.75% from 134°C to 203°C.

Table 7

Example 16: Preparation of Compound A Maleate

Weigh about 8 mg of compound A, dissolve it in 0.08 mL of 2-methyltetrahydrofuran, add an ethanol solution of maleic acid (1 mol/L, 11.89 μL), cycle the temperature from 5°C to 40°C (rate ±0.6°C/min), then add 0.5 mL of n-heptane, stir to precipitate, centrifuge and vacuum dry the solid to obtain a solid.

X-ray powder diffraction analysis revealed no obvious characteristic peaks in the XRPD spectrum, and ion chromatography analysis revealed a maleate ion content of 13.16%.

Example 17: Preparation of Compound A Maleate

Approximately 8 mg of compound A was weighed and dissolved in 0.08 mL of methyl isobutyl ketone. A 1 mol/L ethanolic solution of maleic acid (11.89 μL) was added. The temperature was cycled from 5°C to 40°C (at a rate of ±0.6°C/min). 0.5 mL of n-heptane was then added and stirred to precipitate. After centrifugation, the solid was vacuum-dried to obtain the desired product. X-ray powder diffraction analysis revealed no distinct characteristic peaks in the XRPD spectrum.

Example 18: Preparation of Compound A Maleate

Approximately 8 mg of compound A was weighed and dissolved in 0.08 mL of methanol. A 1 mol/L ethanolic solution of maleic acid (11.89 μL) was added. The temperature was cycled from 5°C to 40°C (at a rate of ±0.6°C/min). 0.5 mL of isopropyl ether was added and stirred to precipitate. After centrifugation, the solid was vacuum-dried to obtain the desired product. X-ray powder diffraction analysis revealed no distinct characteristic peaks in the XRPD spectrum.

Example 19: Preparation of Compound A Citrate

About 8 mg of compound A was weighed and dissolved in 0.08 mL of methyl isobutyl ketone. An ethanol solution of citric acid (1 mol/L, 11.89 μL) was added. The temperature was raised and lowered in a cycle from 5°C to 40°C (rate ±0.6°C/min). The mixture was stirred to precipitate. After centrifugation, the solid was vacuum dried to obtain a solid.

X-ray powder diffraction analysis revealed no obvious characteristic peaks in the XRPD spectrum, and ion chromatography analysis revealed a citrate ion content of 20.60%.

Example 20: Preparation of Compound A Citrate

Approximately 8 mg of compound A was weighed and dissolved in 0.08 mL of 2-methyltetrahydrofuran. A 1 mol/L ethanolic solution of citric acid (11.89 μL) was added. The temperature was cycled from 5°C to 40°C (at a rate of ±0.6°C/min). Stirring was performed to precipitate the product. After centrifugation, the solid was vacuum-dried to obtain the title product. X-ray powder diffraction analysis revealed no distinct characteristic peaks in the XRPD spectrum.

Example 21: Preparation of Compound A Citrate

Approximately 8 mg of compound A was weighed and dissolved in 0.16 mL of methanol. A 1 mol/L ethanolic solution of citric acid (11.89 μL) was added. The temperature was cycled from 5°C to 40°C (at a rate of ±0.6°C/min). 0.5 mL of isopropyl ether was added and stirred to precipitate. After centrifugation, the solid was vacuum dried to obtain the title product. X-ray powder diffraction analysis revealed no distinct characteristic peaks in the XRPD spectrum.

Example 22: Preparation of Compound A Citrate Crystal Form A

Compound A (50 mg, 70.75 μmol) was weighed and dissolved in 1.5 mL of ethyl acetate. The mixture was heated to 50°C and anhydrous citric acid (13.6 mg, 70.8 μmol) was added. The mixture was stirred at 50°C for 2 h, cooled to room temperature and stirred for 16 h. The mixture was filtered and dried in vacuo to obtain a solid (37 mg).

X-ray powder diffraction analysis revealed an XRPD spectrum as shown in Figure 8 , with characteristic peak positions shown in Table 8 , which was defined as citrate crystal form A. Nuclear magnetic resonance (NMR) results showed that the salt ratio of the compound to citric acid was 1:2.

Table 8

Example 23: Preparation of Compound A Citrate Crystal Form B

Compound A (50 mg, 70.75 μmol) was weighed and dissolved in 1.3 mL of acetonitrile. The mixture was heated to 50°C and anhydrous citric acid (13.6 mg, 70.8 μmol) was added. The mixture was stirred at 50°C for 2 h, cooled to room temperature and stirred for 16 h. The mixture was filtered and dried in vacuo to obtain a solid (30 mg).

X-ray powder diffraction analysis revealed an XRPD spectrum as shown in Figure 9, with characteristic peak positions shown in Table 9, which was defined as citrate crystal form B. Nuclear magnetic resonance (NMR) results showed that the salt ratio of the compound to citric acid was 1:1.

Table 9

Example 24: Preparation of Compound A Citrate Crystal Form C

Weigh compound A (50 mg, 70.75 μmol), dissolve it in 1 mL of ethyl acetate, heat to 50°C, add anhydrous citric acid (28.5 mg, 148.3 μmol), stir at 50°C for 2 h, cool to room temperature and continue stirring for 16 h, filter, and dry in vacuo to obtain a solid (55 mg).

X-ray powder diffraction analysis revealed an XRPD spectrum as shown in Figure 10 , with characteristic peak positions shown in Table 10 , defining it as citrate crystal form C. Nuclear magnetic resonance results showed a citric acid content of 29.0%.

Table 10

Example 25: Preparation of Compound A Citrate Crystal Form D

Compound A (50 mg, 70.75 μmol) was weighed and dissolved in 1 mL of ethanol. The mixture was heated to 50°C and anhydrous citric acid (28 mg, 145.7 μmol) was added. The mixture was stirred at 50°C for 2 h, cooled to room temperature and stirred for 16 h. The solid was collected by filtration and dried under vacuum to obtain a solid (46 mg).

X-ray powder diffraction analysis revealed an XRPD spectrum as shown in Figure 11, with characteristic peak positions shown in Table 11, which was defined as citrate crystal form D. Nuclear magnetic resonance (NMR) results showed that the salt ratio of the compound to citric acid was 1:2.

Table 11

Example 26: Preparation of Compound A L-malate

About 8 mg of compound A was weighed and dissolved in 0.16 mL of methanol. An ethanol solution of malic acid (1 mol/L, 11.89 μL) was added, followed by 0.5 mL of isopropyl ether. The temperature was raised and lowered in a cycle from 5°C to 40°C (rate ±0.6°C/min). The mixture was stirred to precipitate. After centrifugation, the solid was vacuum dried to obtain a solid.

X-ray powder diffraction analysis revealed no obvious characteristic peaks in the XRPD spectrum, and ion chromatography analysis revealed a malate ion content of 25.05%.

Example 27: Preparation of p-toluenesulfonate of Compound A

About 8 mg of compound A was weighed and dissolved in 0.08 mL of methyl isobutyl ketone. An ethanol solution of p-toluenesulfonic acid (1 mol/L, 11.89 μL) was added. The temperature was raised and lowered in a cycle from 5°C to 40°C (rate ±0.6°C/min). Then, 0.5 mL of n-heptane was added and stirred to precipitate. After centrifugation, the solid was vacuum dried to obtain a solid.

X-ray powder diffraction analysis revealed no obvious characteristic peaks in the XRPD spectrum, and ion chromatography analysis revealed that the p-toluenesulfonate ion content was 18.42%.

Example 28: Preparation of Compound A Methanesulfonate Crystal Form A

About 8 mg of compound A was weighed and dissolved in 0.08 mL of 2-methyltetrahydrofuran. An ethanol solution of methanesulfonic acid (1 mol/L, 11.89 μL) was added. The temperature was raised and lowered in a cycle from 5°C to 40°C (rate ±0.6°C/min). Then, 0.5 mL of n-heptane was added. The mixture was stirred for crystallization. After centrifugation, the solid was vacuum dried to obtain a solid.

X-ray powder diffraction analysis identified the product as mesylate salt Form A. The XRPD spectrum is shown in Figure 12, and the locations of its characteristic peaks are shown in Table 12. Ion chromatography analysis revealed a mesylate ion content of 14.96%. The DSC spectrum showed endothermic peaks at 178.17°C and 250.19°C, and an exothermic peak at 206.70°C. The TGA spectrum revealed a weight loss of 2.19% between 30°C and 168°C.

Table 12

Example 29: Preparation of Compound A Benzoate Crystal Form A

About 8 mg of compound A was weighed and dissolved in 0.08 mL of methyl isobutyl ketone. An ethanol solution of benzoic acid (1 mol/L, 11.89 μL) was added. The temperature was raised and lowered in a cycle from 5°C to 40°C (rate ±0.6°C/min). The mixture was stirred to precipitate. After centrifugation, the solid was vacuum dried to obtain a solid.

X-ray powder diffraction analysis showed that the product was benzoate crystal form A. The XRPD spectrum is shown in Figure 13, and the positions of its characteristic peaks are shown in Table 13.

Table 13

Example 30: Preparation of Compound A Benzoate Crystal Form A

Weigh about 8 mg of compound A, dissolve it in 0.08 mL of 2-methyltetrahydrofuran, add ethanol solution of benzoic acid (1 mol/L, 11.89 μL), cycle the temperature from 5°C to 40°C (rate ±0.6°C/min), stir to precipitate, centrifuge and vacuum dry the solid to obtain the target product.

Example 31: Preparation of Compound A Succinate Crystal Form A

About 8 mg of compound A was weighed and dissolved in 0.08 mL of methyl isobutyl ketone. A methanol solution of succinic acid (1 mol/L, 11.89 μL) was added. The temperature was raised and lowered in a cycle from 5°C to 40°C (rate ±0.6°C/min). The mixture was stirred for crystallization. After centrifugation, the solid was vacuum dried to obtain a solid.

X-ray powder diffraction analysis identified the product as succinate Form A. The XRPD spectrum is shown in Figure 14, and the locations of its characteristic peaks are shown in Table 14. Ion chromatography analysis revealed a succinate ion content of 13.38%. The DSC spectrum showed an endothermic peak at 175.31°C. The TGA spectrum revealed a weight loss of 1.75% from 30°C to 122°C.

Table 14

Example 32: Preparation of Compound A Succinate Crystal Form B

Compound A (50 mg, 70.7 μmol) was weighed and dissolved in 2 mL of acetonitrile. The temperature was raised to 50°C, succinic acid (8.35 mg, 70.7 μmol) was added, and the mixture was stirred at 50°C for 2 hours. The mixture was cooled to room temperature and stirred for 16 hours. The mixture was filtered and dried in vacuo to obtain a solid (35 mg).

X-ray powder diffraction analysis revealed an XRPD spectrum as shown in Figure 15 and characteristic peak positions as shown in Table 15, defining the product as succinate crystal form B. Nuclear magnetic resonance (NMR) results showed a 1:1 salt ratio of the compound to succinic acid.

Table 15

Example 33: Preparation of Compound A Succinate Crystal Form C

Compound A (50 mg, 70.7 μmol) was weighed and dissolved in 1 mL of ethyl acetate. The temperature was raised to 50°C, and succinic acid (17.5 mg, 148.2 μmol) was added. The mixture was stirred at 50°C for 2 hours, cooled to room temperature, and stirred for 16 hours. The mixture was filtered and dried in vacuo to obtain a solid (38 mg).

The XRPD spectrum of the product is shown in Figure 16 and the characteristic peak positions are shown in Table 16. The product is defined as succinate C crystal form. The NMR results show that the succinic acid content is 25.3%.

Table 16

Example 34: Preparation of Compound A Fumarate

About 8 mg of compound A was weighed and dissolved in 0.16 mL of methanol. A methanol solution of fumaric acid (0.66 mol/L, 18.01 μL) was added, followed by 0.5 mL of isopropyl ether. The temperature was raised and lowered in a cycle from 5°C to 40°C (rate ±0.6°C/min). The mixture was stirred to precipitate. After centrifugation, the solid was vacuum dried to obtain a solid.

X-ray powder diffraction analysis revealed no obvious characteristic peaks in the XRPD spectrum, and ion chromatography analysis revealed a fumarate ion content of 11.40%.

Example 35: Preparation of Compound A Fumarate

Approximately 8 mg of compound A was weighed and dissolved in 0.08 mL of methyl isobutyl ketone. A methanolic solution of fumaric acid (0.66 mol/L, 18.01 μL) was added. The temperature was cycled from 5°C to 40°C (at a rate of ±0.6°C/min). Stirring allowed precipitation to proceed. After centrifugation, the solid was vacuum dried to obtain the title product. X-ray powder diffraction analysis revealed no distinct characteristic peaks in the XRPD spectrum.

Example 36: Preparation of Compound A Fumarate

Approximately 8 mg of compound A was weighed and dissolved in 0.08 mL of 2-methyltetrahydrofuran. A methanolic solution of fumaric acid (0.66 mol/L, 18.01 μL) was added. The temperature was cycled from 5°C to 40°C (at a rate of ±0.6°C/min). Stirring was performed to precipitate the product. After centrifugation, the solid was vacuum-dried to obtain the title product. X-ray powder diffraction analysis revealed no distinct characteristic peaks in the XRPD spectrum.

Example 37: Preparation of Compound A Saccharin Complex

About 8 mg of compound A was weighed and dissolved in 0.16 mL of methanol. 2.18 mg of saccharin solid was added and the temperature was raised and lowered in a cycle from 5°C to 40°C (rate ±0.6°C/min). 0.5 mL of isopropyl ether was added and stirred to precipitate. After centrifugation, the solid was vacuum dried to obtain a solid.

X-ray powder diffraction analysis showed that the XRPD spectrum had no obvious characteristic peaks.

Example 38: Preparation of Compound A Saccharin Complex

Weigh about 8 mg of compound A, dissolve it in 0.16 mL of 2-methyltetrahydrofuran, add 2.18 mg of saccharin solid, cycle the temperature from 5°C to 40°C (rate ±0.6°C/min), add 0.5 mL of n-heptane, stir to precipitate, centrifuge and vacuum dry the solid to obtain the target product.

Example 39: Preparation of Compound A Saccharin Complex

Weigh about 8 mg of compound A, dissolve it in 0.16 mL of methanol, add 2.18 mg of saccharin solid, and heat and cool cyclically from 5°C to 40°C (rate ±0.6°C/min). Add 0.5 mL of n-heptane, stir to precipitate, centrifuge and vacuum dry the solid to obtain the target product.

Test Case 3: Influencing Factors

The above-mentioned salt was spread out in the open and the stability of the samples was examined under high temperature (40°C, 60°C) and high humidity (RH 75%, RH 92.5%) conditions. The sampling period was 30 days.

Table 17

Conclusion: The influencing factor experiment showed that succinate crystal form A showed good physical and chemical stability under high temperature 40℃ and 60℃ and high humidity 75% and 92.5% for 30 days.

Test Example 4: Long-term accelerated test

The aforementioned salts were placed under 25°C/60% RH and 40°C/75% RH conditions to investigate their stability.

Conclusion: Long-term accelerated experiments show that the physicochemical stability of the disclosed crystal forms is good under the conditions of 25°C/60% RH and 40°C/75% RH for 6 months, especially the succinate crystal form A.

Claims (14)

A pharmaceutically acceptable salt of the compound 2-amino-4-((5S,5aS,6S,9R)-12-((1-((4-(difluoromethylidene)piperidin-1-yl)methyl)cyclopropyl)methoxy)-1-fluoro-5-methyl-5a,6,7,8,9,10-hexahydro-5H-4-oxa-3,10a,11,13,14-pentaaza-6,9-methanaphtho[1,8-ab]heptyl-2-yl)-7-fluorobenzo[b]thiophene-3-carbonitrile, characterized in that the pharmaceutically acceptable salt is selected from hydrochloride, sulfate, phosphate, L-tartrate, maleate, citrate, L-malate, p-toluenesulfonate, methanesulfonate, benzoate, succinate and fumarate. The pharmaceutically acceptable salt according to claim 1, characterized in that the chemical ratio of 2-amino-4-((5S,5aS,6S,9R)-12-((1-((4-(difluoromethylidene)piperidin-1-yl)methyl)cyclopropyl)methoxy)-1-fluoro-5-methyl-5a,6,7,8,9,10-hexahydro-5H-4-oxa-3,10a,11,13,14-pentaaza-6,9-methanaphtho[1,8-ab]heptylcyclo-2-yl)-7-fluorobenzo[b]thiophene-3-carbonitrile to the acid molecule is 1:0.5 to 1:3, preferably 1:0.5, 1:1, 1:2 or 1:3. A method for preparing a pharmaceutically acceptable salt according to claim 1 or 2, characterized in that it comprises the step of forming a salt of the compound 2-amino-4-((5S,5aS,6S,9R)-12-((1-((4-(difluoromethylidene)piperidin-1-yl)methyl)cyclopropyl)methoxy)-1-fluoro-5-methyl-5a,6,7,8,9,10-hexahydro-5H-4-oxa-3,10a,11,13,14-pentaaza-6,9-methano[1,8-ab]heptylcyclo-2-yl)-7-fluorobenzo[b]thiophene-3-carbonitrile with an acid. Compound 2-amino-4-((5S,5aS,6S,9R)-12-((1-((4-(difluoromethylidene)piperidin-1-yl)methyl)cyclopropyl)methoxy)-1-fluoro-5-methyl-5a,6,7,8,9,10-hexahydro-5H-4-oxa-3,10a,11,13,14-pentaaza-6,9-methano[1,8-ab]heptyl-2-yl)-7-fluorobenzo[b]thiophene-3-carbonitrile The L-tartrate salt form A is characterized in that the X-ray powder diffraction pattern represented by the diffraction angle 2θ has characteristic peaks at 5.687, 7.991, 10.409, and 12.828, preferably has characteristic peaks at 5.687, 7.991, 10.409, 12.828, 16.244, 20.083, and 21.427, and the most preferred X-ray powder diffraction pattern represented by the diffraction angle 2θ is shown in Figure 1. Compound 2-amino-4-((5S,5aS,6S,9R)-12-((1-((4-(difluoromethylidene)piperidin-1-yl)methyl)cyclopropyl)methoxy)-1-fluoro-5-methyl-5a,6,7,8,9,10-hexahydro-5H-4-oxa-3,10a,11,13,14-pentaaza-6,9-methano[1,8-ab]heptan-2-yl)-7-fluorobenzo[b]thiophene-3-carboxylate The crystalline form A of the methanesulfonate salt of the nitrile is characterized in that the X-ray powder diffraction pattern expressed at a diffraction angle of 2θ has characteristic peaks at 7.553, 9.757, and 13.097, preferably at 7.553, 9.757, 13.097, 16.859, 18.394, 19.738, 20.429, and 21.082. The most preferred X-ray powder diffraction pattern expressed at a diffraction angle of 2θ is shown in Figure 2. Compound 2-amino-4-((5S,5aS,6S,9R)-12-((1-((4-(difluoromethylidene)piperidin-1-yl)methyl)cyclopropyl)methoxy)-1-fluoro-5-methyl-5a,6,7,8,9,10-hexahydro-5H-4-oxa-3,10a,11,13,14-pentaaza-6,9-methano[1,8-ab]heptan-2-yl)-7-fluorobenzo[ b] Thiophene-3-carbonitrile benzoate crystalline form A, characterized in that the X-ray powder diffraction pattern expressed at a diffraction angle of 2θ has characteristic peaks at 7.515, 8.359, and 11.945, preferably at 7.515, 8.002, 8.359, 11.945, 12.920, and 18.970, and most preferably the X-ray powder diffraction pattern expressed at a diffraction angle of 2θ is shown in Figure 3. The compound 2-amino-4-((5S,5aS,6S,9R)-12-((1-((4-(difluoromethylidene)piperidin-1-yl)methyl)cyclopropyl)methoxy)-1-fluoro-5-methyl-5a,6,7,8,9,10-hexahydro-5H-4-oxa-3,10a,11,13,14-pentaaza-6,9-methylnaphtho[1,8-ab]heptyl-2-yl)-7-fluorobenzo[b]thiophene-3-carbonitrile succinate form A is characterized in that the X-ray powder diffraction pattern expressed in terms of diffraction angle 2θ is 7.943, 10.35 1, 12.828, 16.138, 19.832, and 21.057, preferably at 7.943, 10.351, 12.828, 13.212, 16.138, 19.832, 21.057, and 23.619, more preferably at 6.685, 7.943, 10.351, 12.828, 13.212, 16.138, 19.832, 21.057, 21.465, and 23.619, and most preferably the X-ray powder diffraction pattern expressed in terms of diffraction angle 2θ is shown in Figure 4. The crystal form according to any one of claims 4 to 7, characterized in that the 2θ value has an error range of ±0.2. A complex comprising the compound 2-amino-4-((5S,5aS,6S,9R)-12-((1-((4-(difluoromethylidene)piperidin-1-yl)methyl)cyclopropyl)methoxy)-1-fluoro-5-methyl-5a,6,7,8,9,10-hexahydro-5H-4-oxa-3,10a,11,13,14-pentaaza-6,9-methano[1,8-ab]heptyl-2-yl)-7-fluorobenzo[b]thiophene-3-carbonitrile and saccharin. The complex according to claim 9, wherein the chemical ratio of the compound 2-amino-4-((5S,5aS,6S,9R)-12-((1-((4-(difluoromethylidene)piperidin-1-yl)methyl)cyclopropyl)methoxy)-1-fluoro-5-methyl-5a,6,7,8,9,10-hexahydro-5H-4-oxa-3,10a,11,13,14-pentaaza-6,9-methylnaphtho[1,8-ab]heptylcyclo-2-yl)-7-fluorobenzo[b]thiophene-3-carbonitrile to the saccharin molecule is 1:1. A pharmaceutical composition comprising the pharmaceutically acceptable salt according to claim 1 or 2, or the crystal form according to claims 4-8, or the complex according to claim 9 or 10, and a pharmaceutically acceptable excipient. A pharmaceutical composition is prepared from the pharmaceutically acceptable salt according to claim 1 or 2, or the crystal form according to claims 4-8, or the complex according to claim 9 or 10, and a pharmaceutically acceptable excipient. Use of the pharmaceutically acceptable salt of claim 1 or 2, the crystalline form of claims 4-8, or the complex of claim 9 or 10, or the pharmaceutical composition of claim 11 or 12 in the preparation of a medicament for preventing and/or treating a disease or condition mediated by KRAS G12D. Use of the pharmaceutically acceptable salt of claim 1 or 2, the crystalline form of claim 4-8, or the complex of claim 9 or 10, or the pharmaceutical composition of claim 11 or 12 in the preparation of a medicament for preventing and/or treating tumors, wherein the tumor is preferably selected from brain cancer, thyroid cancer, head and neck cancer, nasopharyngeal cancer, pharyngeal cancer, oral cancer, salivary gland cancer, esophageal cancer, gastric cancer, lung cancer, liver cancer, kidney cancer, pancreatic cancer, gallbladder cancer, bile duct cancer, colorectal cancer, small intestine cancer, gastrointestinal stromal tumor, urothelial carcinoma, urethral cancer, bladder cancer, breast cancer, vaginal cancer, ovarian cancer, endometrial cancer, cervical cancer, fallopian tube cancer, testicular cancer, prostate cancer, hemangioma, leukemia, lymphoma, myeloma, skin cancer, lipoma, bone cancer, soft tissue sarcoma, neurofibroma, glioma, neuroblastoma and glioblastoma; more preferably selected from pancreatic cancer, colorectal cancer and non-small cell lung cancer.