CN113493440A - Salt of nitrogen-containing heteroaromatic derivative and crystal form thereof - Google Patents

Salt of nitrogen-containing heteroaromatic derivative and crystal form thereof Download PDF

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CN113493440A
CN113493440A CN202110360835.8A CN202110360835A CN113493440A CN 113493440 A CN113493440 A CN 113493440A CN 202110360835 A CN202110360835 A CN 202110360835A CN 113493440 A CN113493440 A CN 113493440A
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詹小兰
呙临松
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Jiangsu Hansoh Pharmaceutical Group Co Ltd
Shanghai Hansoh Biomedical Co Ltd
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Abstract

The invention relates to a salt of a nitrogen-containing heteroaromatic derivative and a crystal form thereof. In particular to a salt of a compound with a general formula (I), a crystal form and a preparation method thereof, a pharmaceutical composition containing a treatment and effective amount of the salt of the derivative and the crystal form thereof, and a casein prepared from the saltUse of an inhibitor of the enzyme, the enzyme-2C (SHP2), for the treatment of diseases or conditions selected from leukemia, neuroblastoma, melanoma, breast cancer, lung cancer and colon cancer.
Figure DDA0003005435290000011

Description

Salt of nitrogen-containing heteroaromatic derivative and crystal form thereof
Technical Field
The invention belongs to the field of drug synthesis, and particularly relates to salts of nitrogen-containing heteroaromatic derivatives, a crystal form of the salts, a preparation method of the salts and an application of the salts.
Background
SHP-2(Src homology-2 domain-associating phosphatase 2), also known as PTPN11 (type-protein phosphatase non-receptor type 11), is encoded by the PTPN11 gene and belongs to the family of Protein Tyrosine Phosphatases (PTPs). As downstream signal molecules of cytokines, growth factors and other extracellular stimulators, SHP-2 is widely expressed in various tissues and cells of the body, and is involved in cell signal transduction, regulation of cell growth, differentiation, migration, metabolism, gene transcription, immune response and the like.
SHP-2 has mainly 3 components: SH-2 domains (N-SH2 and C-SH2), PTP-active domains, the C-terminus (containing tyrosine phosphorylation sites). Among them, the SH2 domain is highly conserved, is a phosphotyrosine binding site, and mediates the binding of the PTP domain to its ligand.
SHP-2 has 2 main states in the body: and (4) inactivation and activation. In the inactivated state, the N-SH2 inside SHP-2 is combined with PTP domain mutually, and SHP-2 is inactivated because the PTP domain is occupied. When N-SH2 is specifically bound with the phosphorylated tyrosine residue ligand, the PTP domain is re-exposed, and the SHP-2 restores the activity. Recent studies have shown that SHP-2 is also able to form dimers in vivo, and that it also leads to inactivation of SHP-2.
SHP-2 mainly plays a role by regulating signal pathways such as ERK/MAPK, JAK-STAT, PI3K/AKT, Hippo, Wnt/beta-catenin and the like, thereby maintaining the development and homeostasis of organisms. Specific studies have shown that SHP-2 participates in activation of the ERK/MAPK pathway by directly binding to tyrosine kinase Receptors (RTKs) or scaffold. In addition, activated SHP-2 recruits GRB2/SOS, indirectly facilitating activation of the RAS signaling pathway. In addition, SHP-2 is also involved in suppressing signaling of immune responses, e.g., SHP-2 and SHP-1 are able to bind and activate immunosuppressive receptors (e.g., PD-1), blocking T cell activation.
As an important class of cell signaling factors, SHP-2 mutations are closely associated with a variety of diseases. The research finds that: SHP-2 mutations are found in neuroblastoma, AML (4%), breast cancer, NSCLC (10%), lung adenocarcinoma (30%), esophageal cancer, head and neck tumors, melanoma, and gastric cancer.
The mutation site of SHP-2 mostly occurs in N-SH2 and PTP active region, which removes the mutual inhibition of N-CH2/PTP structural domain, and generates high-activity SHP-2, and mutations such as Cys459Ser mutant and E76K mutant affect the activity of SHP-2. Research shows that the high-activity SHP-2 is closely related to inflammation, liver cirrhosis, toxin CagA secreted by helicobacter pylori and the like, can cause the regeneration and development of tumors, and is equivalent to protooncogenes. With the continuous and deep knowledge of SHP-2, SHP-2 has been used as a target for tumor therapy for drug development.
Several allosteric inhibitors of SHP-2 have been introduced into clinical research, such as TNO-155 developed by Novartis, RMC-4630 developed by Revolition Medicine, and JAB-3068 by Beijing plus Corse. However, no SHP-2 inhibitor has been developed and marketed for the treatment of Noonan syndrome, leopard syndrome, leukemia, neuroblastoma, melanoma, breast cancer, esophageal cancer, head and neck tumors, lung cancer and colon cancer. Therefore, the development of SHP-2 inhibitor drugs with good pharmacy is urgently needed.
The invention discloses a series of structures of nitrogen-containing heteroaromatic derivative inhibitors in PCT patent (application number: PCT/CN 2019/110314). in the subsequent development, in order to achieve the characteristics of easiness in processing, filtering and drying products, suitability for storage, long-term stability of products, high bioavailability and the like, the invention comprehensively researches different acid salts of free bases of the substances, and aims to obtain the most suitable crystal form.
Disclosure of Invention
All that is referred to in patent PCT/CN2019/110314 is added to the present invention by way of reference.
The invention aims to provide an acid salt of a compound shown as a general formula (I), which has the following structure:
Figure BDA0003005435270000021
wherein:
R1selected from hydrogen, halogen, C1-6Alkyl radical, C1-6Alkoxy radical, C1-6Hydroxyalkyl radical, C3-8Cycloalkyl, 3-12 membered heterocyclyl, C6-12Aryl or 5-12 membered heteroaryl, wherein said C1-6Alkyl radical, C1-6Alkoxy radical, C1-6Hydroxyalkyl radical, C3-8Cycloalkyl, 3-12 membered heterocyclyl, C6-12Aryl and 5-12 membered heteroaryl, optionally further substituted by deuterium, halogen, amino, nitro, hydroxy, cyano, oxo, C1-6Alkyl radical, C1-6Deuterated alkyl, C1-6Haloalkyl, C1-6Alkoxy radical, C1-6Hydroxyalkyl radical, C1-6Haloalkoxy, C2-6Alkenyl radical, C2-6Alkynyl, C3-8Cycloalkyl, 3-12 membered heterocyclyl, C 6-12Aryl and 5-12 membered heteroaryl;
R2selected from hydrogen, amino or C1-6An alkyl group;
R3selected from hydrogen, halogen, amino, cyano, C1-6Alkyl radical, C1-6Alkoxy radical, C1-6Hydroxyalkyl radical, C3-8Cycloalkyl or 3-12 membered heterocyclyl, wherein said C is1-6Alkyl radical, C1-6Alkoxy radical, C1-6Hydroxyalkyl radical, C3-8Cycloalkyl and 3-12 membered heterocyclyl, optionally further deuterium,Halogen, amino, nitro, hydroxy, cyano, oxo, C1-6Alkyl radical, C1-6Deuterated alkyl, C1-6Haloalkyl, C1-6Alkoxy radical, C1-6Hydroxyalkyl radical, C1-6Haloalkoxy, C2-6Alkenyl radical, C2-6Alkynyl, C3-8Cycloalkyl, 3-12 membered heterocyclyl, C6-12Aryl and 5-12 membered heteroaryl;
R4selected from hydrogen, deuterium, halogen, amino, nitro, hydroxy, cyano, C1-6Alkyl radical, C1-6Deuterated alkyl, C1-6Haloalkyl, C1-6Alkoxy radical, C1-6Hydroxyalkyl radical, C1-6Haloalkoxy, C3-8Cycloalkyl, 3-12 membered heterocyclyl, C6-12Aryl or 5-12 membered heteroaryl;
R5and R6Each independently selected from hydrogen, deuterium, halogen, amino, nitro, hydroxy, cyano, C1-6Alkyl radical, C1-6Deuterated alkyl, C1-6Haloalkyl, C1-6Alkoxy radical, C1-6Haloalkoxy, C1-6Hydroxyalkyl radical, C2-6Alkenyl radical, C3-8Cycloalkyl, 3-12 membered heterocyclyl, C 6-12Aryl or 5-12 membered heteroaryl, wherein said C1-6Alkyl radical, C1-6Deuterated alkyl, C1-6Haloalkyl, C1-6Alkoxy radical, C1-6Haloalkoxy, C1-6Hydroxyalkyl radical, C2-6Alkenyl radical, C3-8Cycloalkyl, 3-12 membered heterocyclyl, C6-12Aryl and 5-12 membered heteroaryl, optionally further substituted by deuterium, halogen, amino, nitro, hydroxy, cyano, oxo, C1-6Alkyl radical, C1-6Deuterated alkyl, C1-6Haloalkyl, C1-6Alkoxy radical, C1-6Hydroxyalkyl radical, C1-6Haloalkoxy, C2-6Alkenyl radical, C2-6Alkynyl, C3-8Cycloalkyl, 3-12 membered heterocyclyl, C6-12Aryl and 5-12 membered heteroaryl;
or, R5And R6Are linked to form a C3-8CycloalkanesA group, a 3-12 membered heterocyclic group, C6-12Aryl or 5-12 membered heteroaryl, wherein said C3-8Cycloalkyl, 3-12 membered heterocyclyl, C6-12Aryl and 5-12 membered heteroaryl, optionally further substituted by one or more substituents selected from deuterium, halogen, amino, nitro, hydroxy, cyano, oxo, C1-6Alkyl radical, C1-6Deuterated alkyl, C1-6Haloalkyl, C1-6Alkoxy radical, C1-6Hydroxyalkyl radical, C1-6Haloalkoxy, C2-6Alkenyl radical, C2-6Alkynyl, C3-8Cycloalkyl, 3-12 membered heterocyclyl, C6-12Aryl and 5-12 membered heteroaryl;
m is an inorganic or organic acid, wherein the inorganic acid is selected from hydrochloric acid, sulfuric acid, nitric acid, hydrobromic acid, hydrofluoric acid, hydroiodic acid, or phosphoric acid; the organic acid is selected from 2, 5-dihydroxybenzoic acid, 1-hydroxy-2-naphthoic acid, acetic acid, dichloroacetic acid, trichloroacetic acid, acetoxy-hydroxamic acid, adipic acid, benzenesulfonic acid, 4-chlorobenzenesulfonic acid, benzoic acid, 4-acetamidobenzoic acid, 4-aminobenzoic acid, capric acid, caproic acid, caprylic acid, cinnamic acid, citric acid, cyclamic acid, camphorsulfonic acid, aspartic acid, camphoric acid, gluconic acid, glucuronic acid, glutamic acid, isoascorbic acid, lactic acid, malic acid, mandelic acid, pyroglutamic acid, tartaric acid, dodecylsulfuric acid, dibenzoyltartaric acid, ethane-1, 2-disulfonic acid, ethanesulfonic acid, formic acid, fumaric acid, galactaric acid, gentisic acid, glutaric acid, 2-ketoglutaric acid, glycolic acid, hippuric acid, isethionic acid, lactobionic acid, lactic acid, tartaric acid, citric acid, tartaric acid, citric acid, tartaric acid, citric acid, tartaric acid, citric acid, tartaric acid, citric acid, ascorbic acid, aspartic acid, lauric acid, camphoric acid, maleic acid, malonic acid, methanesulfonic acid, 1, 5-naphthalenedisulfonic acid, naphthalene-2-sulfonic acid, nicotinic acid, oleic acid, orotic acid, oxalic acid, palmitic acid, pamoic acid, propionic acid, salicylic acid, 4-aminosalicylic acid, sebacic acid, stearic acid, succinic acid, thiocyanic acid, undecylenic acid, trifluoroacetic acid, benzenesulfonic acid, p-toluenesulfonic acid, or L-malic acid; and is
y is 1, 2 or 3.
In a further preferred embodiment of the invention, R1Selected from hydrogen, halogen, C1-6Alkyl radical, C1-6Hydroxyalkyl radical, C3-8Cycloalkyl radicalsOr 3-12 membered heterocyclic group, wherein said C is1-6Alkyl radical, C1-6Hydroxyalkyl radical, C3-8Cycloalkyl and 3-12 membered heterocyclyl, optionally further substituted by deuterium, halogen, hydroxy, oxo, C1-6Alkyl and C3-8Cycloalkyl substituted with one or more substituents; more preferably hydrogen, halogen, C1-3Alkyl radical, C1-3Hydroxyalkyl radical, C3-6Cycloalkyl or 3-6 membered heterocyclyl, wherein said C is1-3Alkyl radical, C1-3Hydroxyalkyl radical, C3-6Cycloalkyl and 3-6 membered heterocyclyl, optionally further substituted by deuterium, halogen, hydroxy, oxo, C1-3Alkyl and C3-6Cycloalkyl substituted with one or more substituents; further preferred are hydrogen, fluorine, bromine, methyl, hydroxyethyl,
Figure BDA0003005435270000041
Figure BDA0003005435270000042
In a further preferred embodiment of the invention, R2Selected from hydrogen, amino or C1-6An alkyl group; preferably hydrogen, amino or C1-3An alkyl group; more preferably hydrogen, amino, methyl, ethyl or propyl; further preferred is hydrogen, amino or methyl.
In a further preferred embodiment of the invention, R3Selected from hydrogen, halogen, amino, C1-6Alkyl radical, C3-8Cycloalkyl or 3-12 membered heterocyclyl, wherein said C is1-6Alkyl radical, C3-8Cycloalkyl and 3-12 membered heterocyclyl, optionally further substituted with one or more substituents of deuterium, halogen, oxo; preferably hydrogen, halogen, amino, C 1-3Alkyl radical, C3-6Cycloalkyl or 3-6 membered heterocyclyl, wherein said C is1-3Alkyl radical, C3-6Cycloalkyl and 3-6 membered heterocyclyl, optionally further substituted with one or more substituents of deuterium, halogen, oxo; further preferred are amino, chloro, cyclopropyl, azetidinyl, tetrahydropyrrolyl, morpholinyl, azetidinyl, pyrrolyl, morpholinyl, pyrrolyl, and thienyl,
Figure BDA0003005435270000043
Figure BDA0003005435270000044
In a further preferred embodiment of the invention, R4Selected from hydrogen, halogen, C1-6Alkyl or C3-8A cycloalkyl group; preferably hydrogen, halogen, C1-3Alkyl or C3-6A cycloalkyl group; further preferred is hydrogen, fluorine, chlorine, methyl or cyclopropyl.
In a further preferred embodiment of the invention, R5And R6Each independently selected from hydrogen and C1-6Alkyl radical, C1-6Haloalkyl, C2-6Alkenyl or C3-8Cycloalkyl, wherein said C1-6Alkyl radical, C1-6Haloalkyl, C2-6Alkenyl and C3-8Cycloalkyl optionally further substituted by deuterium, halogen, C1-6Substituted by one or more substituents in the alkyl group; preferably hydrogen, C1-3Alkyl radical, C2-4Alkenyl or C3-6Cycloalkyl, wherein said C1-3Alkyl radical, C2-4Alkenyl and C3-6Cycloalkyl optionally further substituted by deuterium, halogen, C1-3Substituted by one or more substituents in the alkyl group; further preferred are hydrogen, methyl, ethyl, isopropyl, vinyl, cyclopropyl,
Figure BDA0003005435270000045
Or, R 5And R6Are linked to form a C3-8Cycloalkyl, 6-12 aryl or 5-12 membered heteroaryl, wherein said C is3-8Cycloalkyl, 6-12 aryl and 5-12 membered heteroaryl, optionally further substituted by one or more substituents selected from deuterium, halogen or C1-6Substituted by one or more substituents in the alkyl group; preferably C3-6Cycloalkyl, phenyl or 5-6 membered heteroaryl, wherein said C3-6Cycloalkyl, phenyl and 5-6 membered heteroaryl, optionally further substituted by one or more substituents selected from deuterium, halogen or C1-3Substituted by one or more substituents in the alkyl group; further preferred is
Figure BDA0003005435270000051
Figure BDA0003005435270000052
In a further preferred embodiment of the invention, M is selected from hydrochloric acid, phosphoric acid, acetic acid, benzoic acid, p-toluenesulfonic acid, succinic acid, tartaric acid, ethanesulfonic acid, benzenesulfonic acid, fumaric acid, isethionic acid, oxalic acid, hydrobromic acid or gentisic acid; preferably hydrochloric acid, ethanesulfonic acid, benzenesulfonic acid, fumaric acid, isethionic acid, oxalic acid, hydrobromic acid or gentisic acid; further preferred is hydrochloric acid or ethanesulfonic acid; and is
y is 1 or 2.
In a further preferred embodiment of the present invention, the compound has the structure according to formula (II):
Figure BDA0003005435270000053
wherein:
ring A is selected from C3-8Cycloalkyl, 3-12 membered heterocyclyl, C6-10Aryl or 5-12 membered heteroaryl;
Raselected from hydrogen, deuterium, halogen, amino, nitro, hydroxy, cyano, C 1-6Alkyl radical, C1-6Deuterated alkyl, C1-6Haloalkyl, C1-6Alkoxy radical, C1-6Hydroxyalkyl radical, C1-6Haloalkoxy, C2-6Alkenyl radical, C2-6Alkynyl, C3-8Cycloalkyl, 3-12 membered heterocyclyl, C6-12Aryl or 5-12 membered heteroaryl; and is
x is 0, 1, 2 or 3.
In a further preferred embodiment of the invention, the ring a is phenyl.
In a further preferred embodiment of the present invention, an acid salt, preferably a hydrochloride, ethanesulfonate, benzenesulfonate, fumarate, isethionate, oxalate, hydrobromide or gentisate, of the compound (S) -1'- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) -3-fluoropyrazin-2-yl) -1, 3-dihydrospiro [ indene-2, 4' -piperidin ] -1-amine is represented by formula (III):
Figure BDA0003005435270000054
in a further preferred embodiment of the invention, the compound (S) -1'- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) -3-fluoropyrazin-2-yl) -1, 3-dihydrospiro [ indene-2, 4' -piperidin ] -1-amine hydrochloride.
In a further preferred embodiment of the invention, the compound (S) -1'- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) -3-fluoropyrazin-2-yl) -1, 3-dihydrospiro [ indene-2, 4' -piperidine ] -1-amine is an ethanesulfonic acid salt.
In a further preferred embodiment of the invention, the compound (S) -1'- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) -3-fluoropyrazin-2-yl) -1, 3-dihydrospiro [ indene-2, 4' -piperidine ] -1-amine is a benzenesulfonate salt.
In a further preferred embodiment of the invention, the compound (S) -1'- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) -3-fluoropyrazin-2-yl) -1, 3-dihydrospiro [ indene-2, 4' -piperidin ] -1-amine is the fumarate salt.
In a further preferred embodiment of the invention, the compound (S) -1'- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) -3-fluoropyrazin-2-yl) -1, 3-dihydrospiro [ indene-2, 4' -piperidine ] -1-amine is the isethionate salt.
In a further preferred embodiment of the invention, the oxalate salt of the compound (S) -1'- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) -3-fluoropyrazin-2-yl) -1, 3-dihydrospiro [ indene-2, 4' -piperidine ] -1-amine.
In a further preferred embodiment of the present invention, the compound (S) -1'- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) -3-fluoropyrazin-2-yl) -1, 3-dihydrospiro [ indene-2, 4' -piperidin ] -1-amine is a hydrobromide salt.
In a further preferred embodiment of the invention, the compound (S) -1'- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) -3-fluoropyrazin-2-yl) -1, 3-dihydrospiro [ indene-2, 4' -piperidine ] -1-amine is gentisate.
In a further preferred embodiment of the invention, the number of acids in the acid salt is 1 to 3, preferably 1, 1.5, 2, 2.5 or 3, more preferably 1 or 2.
The invention further provides a method for preparing the acid salt, which comprises the following steps:
1) weighing a proper amount of compound free alkali, and dissolving the compound free alkali by using a benign solvent;
2) dissolving the acid M in an organic solvent; the amount of acid M is preferably 2 equivalents;
3) combining the two solutions obtained in the steps 1) and 2) to obtain a salt solution, and stirring and precipitating the salt solution or dropwise adding a poor solvent and stirring and precipitating the solution;
4) quickly centrifuging or standing and drying to obtain a target product;
wherein:
the benign solvent of the step 1) is selected from methanol, ethanol, ethyl acetate, dichloromethane, acetone, acetonitrile, 2-butanone, 3-pentanone, tetrahydrofuran, N-dimethylformamide, N-dimethylacetamide or N-methylpyrrolidone; preferably methanol or ethanol;
the organic solvent in the step 2) is selected from methanol, ethanol, ethyl acetate, dichloromethane, acetone, N-hexane, petroleum ether, benzene, toluene, chloroform, acetonitrile, carbon tetrachloride, dichloroethane, tetrahydrofuran, 2-butanone, 3-pentanone, heptane, methyl tert-butyl ether, isopropyl ether, 1, 4-dioxane, tert-butyl alcohol or N, N-dimethylformamide; preferably methanol, ethanol or acetonitrile;
the benign solvent in the step 1) and the organic solution in the step 2) need to be mutually soluble when being used;
The poor solvent of the step 3) is selected from heptane, water, methyl tert-butyl ether, toluene, isopropyl ether, ethyl acetate, acetone or acetonitrile; water, methyl tert-butyl ether or isopropyl ether are preferred.
Alternatively, the first and second electrodes may be,
1) weighing a proper amount of free alkali, dissolving the free alkali in a benign solvent,
2) weighing a proper amount of acid, and dissolving the acid by using an organic solvent; the amount of acid is preferably 1.2 equivalents,
3) mixing the two solutions, stirring to separate out or dripping poor solvent and stirring to separate out,
4) drying the mixture to obtain a target product,
wherein:
the benign solvent is selected from methanol, dichloromethane, 1, 4-dioxane, acetone, tetrahydrofuran, N-dimethylformamide, N-dimethylacetamide or N-methylpyrrolidone; preferably methanol or tetrahydrofuran, and preferably, methanol or tetrahydrofuran,
the organic solvent is selected from methanol, ethanol, ethyl acetate, dichloromethane, acetone, N-hexane, petroleum ether, benzene, toluene, chloroform, acetonitrile, carbon tetrachloride, dichloroethane, tetrahydrofuran, 2-butanone, 3-pentanone, heptane, methyl tert-butyl ether, isopropyl ether, 1, 4-dioxane, tert-butanol or N, N-dimethylformamide; preferably methanol, ethanol or acetonitrile; the benign solvent and the organic solution need to be mutually soluble when in use,
the poor solvent is selected from heptane, water, methyl tert-butyl ether, toluene, isopropyl ether, ethyl acetate, acetone or acetonitrile; preferably water, methyl tert-butyl ether, isopropyl ether;
Alternatively, the first and second electrodes may be,
1) weighing a proper amount of free alkali, suspending with a poor solvent 1,
2) weighing a proper amount of acid, and dissolving the acid by using an organic solvent; the amount of acid is preferably 1.2 equivalents,
3) mixing the two solutions, stirring for dissolving, continuously stirring for precipitating or dripping the poor solvent 2 and stirring for precipitating,
4) drying the mixture to obtain a target product,
the poor solvent 1 is selected from ethyl acetate, acetone, dichloromethane, acetonitrile, tetrahydrofuran, 2-methyl-tetrahydrofuran, 2-butanone, 3-pentanone, 1, 4-dioxane and ethanol; preferably ethyl acetate, acetone, acetonitrile, 2-methyl-tetrahydrofuran, ethanol,
the organic solvent is selected from methanol, ethanol, ethyl acetate, dichloromethane, acetone, N-hexane, petroleum ether, benzene, toluene, chloroform, acetonitrile, carbon tetrachloride, dichloroethane, tetrahydrofuran, 2-butanone, 3-pentanone, heptane, methyl tert-butyl ether, isopropyl ether, 1, 4-dioxane, tert-butanol or N, N-dimethylformamide; preferably methanol, ethanol or acetonitrile; the benign solvent and the organic solution need to be mutually soluble when in use,
the poor solvent 2 is selected from heptane, water, methyl tert-butyl ether, toluene, isopropyl ether, ethyl acetate, acetone or acetonitrile; preferably water, methyl tert-butyl ether, isopropyl ether;
The acid is selected from hydrochloric acid, sulfuric acid, nitric acid, hydrobromic acid, hydrofluoric acid, hydroiodic acid, phosphoric acid, 2, 5-dihydroxybenzoic acid, 1-hydroxy-2-naphthoic acid, acetic acid, dichloroacetic acid, trichloroacetic acid, acetoxy-hydroxamic acid, adipic acid, benzenesulfonic acid, 4-chlorobenzenesulfonic acid, benzoic acid, 4-acetamidobenzoic acid, 4-aminobenzoic acid, capric acid, caproic acid, caprylic acid, cinnamic acid, citric acid, cyclamic acid, camphorsulfonic acid, aspartic acid, camphoric acid, gluconic acid, glucuronic acid, glutamic acid, isoascorbic acid, lactic acid, malic acid, mandelic acid, pyroglutamic acid, tartaric acid, dodecylsulfuric acid, dibenzoyltartaric acid, ethane-1, 2-disulfonic acid, ethanesulfonic acid, formic acid, fumaric acid, galactaric acid, gentisic acid, glutaric acid, 2-ketoglutaric acid, Glycolic acid, hippuric acid, isethionic acid, lactobionic acid, ascorbic acid, aspartic acid, lauric acid, camphoric acid, maleic acid, malonic acid, methanesulfonic acid, 1, 5-naphthalenedisulfonic acid, naphthalene-2-sulfonic acid, nicotinic acid, oleic acid, orotic acid, oxalic acid, palmitic acid, pamoic acid, propionic acid, salicylic acid, 4-aminosalicylic acid, sebacic acid, stearic acid, succinic acid, thiocyanic acid, undecylenic acid, trifluoroacetic acid, benzenesulfonic acid, p-toluenesulfonic acid, or L-malic acid; preferably fumaric acid, p-toluenesulfonic acid or succinic acid; most preferred is p-toluenesulfonic acid.
In a further preferred embodiment of the present invention, said formula (I) is crystalline or amorphous.
In a further preferred embodiment of the present invention, the compound of formula (I) is a hydrate or an anhydrate, preferably an anhydrate.
In a further preferred embodiment of the present invention, the compound (S) -1'- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) -3-fluoropyrazin-2-yl) -1, 3-dihydrospiro [ indene-2, 4' -piperidin ] -1-amine is in the form of an acid salt, preferably in the form of a hydrochloride, ethanesulfonate, benzenesulfonate, fumarate, isethionate, oxalate, hydrobromide or gentisate salt.
In a further preferred embodiment of the present invention, the compound (S) -1'- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) -3-fluoropyrazin-2-yl) -1, 3-dihydrospiro [ indene-2, 4' -piperidin ] -1-amine is in the form of its hydrochloride salt.
In a further preferred embodiment of the invention, the compound (S) -1'- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) -3-fluoropyrazin-2-yl) -1, 3-dihydrospiro [ indene-2, 4' -piperidine ] -1-amine is in the form of its ethanesulfonate salt.
In a further preferred embodiment of the invention, the compound (S) -1'- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) -3-fluoropyrazin-2-yl) -1, 3-dihydrospiro [ indene-2, 4' -piperidine ] -1-amine is in the form of its benzenesulfonate salt in crystalline form.
In a further preferred embodiment of the present invention, the compound (S) -1'- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) -3-fluoropyrazin-2-yl) -1, 3-dihydrospiro [ indene-2, 4' -piperidin ] -1-amine is in the form of its fumarate salt.
In a further preferred embodiment of the invention, the compound (S) -1'- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) -3-fluoropyrazin-2-yl) -1, 3-dihydrospiro [ indene-2, 4' -piperidine ] -1-amine is in crystalline form as isethionate.
In a further preferred embodiment of the invention, the compound (S) -1'- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) -3-fluoropyrazin-2-yl) -1, 3-dihydrospiro [ indene-2, 4' -piperidine ] -1-amine is in the form of its oxalate salt.
In a further preferred embodiment of the present invention, the compound (S) -1'- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) -3-fluoropyrazin-2-yl) -1, 3-dihydrospiro [ indene-2, 4' -piperidin ] -1-amine is in the form of its hydrobromide salt.
In a further preferred embodiment of the invention, the compound (S) -1'- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) -3-fluoropyrazin-2-yl) -1, 3-dihydrospiro [ indene-2, 4' -piperidine ] -1-amine is in crystalline form as gentisate salt.
In a further preferred embodiment of the invention, in the crystalline form of the acid salt, the number of acids is from 1 to 3, preferably 1, 1.5, 2, 2.5 or 3.
In a further preferred embodiment of the present invention, the acid salt of the compound (S) -8- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) pyrazin-2-yl) -2-cyclopropyl-8-azaspiro [4.5] dec-2-en-1-amine, preferably methanesulfonate, isethionate, fumarate, p-toluenesulfonate, ethanesulfonate, succinate, hydrobromide, hydrochloride, maleate, tartrate or sulfate,
the number of the p-toluenesulfonate salt-forming acids is 1, the number of the fumarate salt-forming acids is 0.5, and the number of the succinate salt-forming acids is 0.5.
In a further preferred embodiment of the present invention, the compound (S) -8- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) pyrazin-2-yl) -2-cyclopropyl-8-azaspiro [4.5] dec-2-en-1-amine is in the form of its mesylate salt.
In a further preferred embodiment of the invention, the compound (S) -8- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) pyrazin-2-yl) -2-cyclopropyl-8-azaspiro [4.5] dec-2-en-1-amine is in the form of its isethionate salt.
In a further preferred embodiment of the present invention, the compound (S) -8- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) pyrazin-2-yl) -2-cyclopropyl-8-azaspiro [4.5] dec-2-en-1-amine is in the form of the fumarate salt.
In a further preferred embodiment of the present invention, the compound (S) -8- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) pyrazin-2-yl) -2-cyclopropyl-8-azaspiro [4.5] dec-2-en-1-amine is in crystalline form as p-toluenesulfonate.
In a further preferred embodiment of the present invention, the compound (S) -8- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) pyrazin-2-yl) -2-cyclopropyl-8-azaspiro [4.5] dec-2-en-1-amine is in the form of its ethanesulfonate salt.
In a further preferred embodiment of the invention, the compound (S) -8- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) pyrazin-2-yl) -2-cyclopropyl-8-azaspiro [4.5] dec-2-en-1-amine is in the form of the succinate salt.
In a further preferred embodiment of the present invention, the compound (S) -8- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) pyrazin-2-yl) -2-cyclopropyl-8-azaspiro [4.5] dec-2-en-1-amine is in the form of its hydrobromide salt.
In a further preferred embodiment of the invention, the compound (S) -8- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) pyrazin-2-yl) -2-cyclopropyl-8-azaspiro [4.5] dec-2-en-1-amine is in the form of its hydrochloride salt.
In a further preferred embodiment of the present invention, the compound (S) -8- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) pyrazin-2-yl) -2-cyclopropyl-8-azaspiro [4.5] dec-2-en-1-amine is in the crystalline maleate form.
In a further preferred embodiment of the invention, the compound (S) -8- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) pyrazin-2-yl) -2-cyclopropyl-8-azaspiro [4.5] dec-2-en-1-amine is in the form of its tartrate salt.
In a further preferred embodiment of the present invention, the compound (S) -8- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) pyrazin-2-yl) -2-cyclopropyl-8-azaspiro [4.5] dec-2-en-1-amine is in the form of the sulfate salt.
In a further preferred embodiment of the present invention, the compound (S) -1'- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) -3-fluoropyrazin-2-yl) -1, 3-dihydrospiro [ indene-2, 4' -piperidin ] -1-amine is in the hydrochloride form a in the form of hydrochloride salt with a number of 1 or 2, preferably the salt formation ratio of the compound of formula (III) to ethanesulfonic acid is 1: 1. ESLD detection analysis shows that the mole ratio of free alkali to chloride ion content in hydrochloric acid is 1:1, the salt formation ratio of (S) -1'- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) -3-fluoropyrazin-2-yl) -1, 3-dihydrospiro [ indene-2, 4' -piperidine ] -1-amine free base to hydrochloric acid can be seen to be 1: 1.
Wherein the X-ray powder diffraction pattern has diffraction peaks at 2 theta of 15.1 degrees, 21.0 degrees, 22.6 degrees and 23.7 degrees; further comprising diffraction peaks at 8.4 °, 9.4 °, 16.7 °, 18.4 °, and 25.5 ° 2 θ; further comprising diffraction peaks at 14.5 °, 19.2 °, 20.5 °, 24.8 °, 25.1 °, 27.8 °, and 31.4 ° 2 θ; still further comprising diffraction peaks at 11.3 °, 16.3 °, 23.4 °, 24.1 °, 26.4 °, 28.2 °, 29.0 °, 30.6 °, 32.3 °, 32.8 °, 35.6 °, and 37.0 ° of 2 θ. Most preferably, the XRPD pattern is substantially as shown in figure 1.
Characteristic diffraction peaks of X-rays expressed in terms of 2 theta angle and interplanar spacing d using Cu-K.alpha.radiation are shown in Table 1.
TABLE 1
Figure BDA0003005435270000101
Figure BDA0003005435270000111
The compound (S) -1'- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) -3-fluoropyrazin-2-yl) -1, 3-dihydrospiro [ indene-2, 4' -piperidine ] -1-amine hydrochloride crystal form A has a DSC pattern basically as shown in figure 2; the TGA profile is substantially as shown in figure 3.
In a preferred embodiment of the present invention, the compound (S) -1'- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) -3-fluoropyrazin-2-yl) -1, 3-dihydrospiro [ indene-2, 4' -piperidin ] -1-amine is in the form of its hydrochloride in crystalline form B, said number of hydrochloric acids being 1 or 2, preferably 1.
Wherein the X-ray powder diffraction pattern has diffraction peaks at 2 theta of 6.9 degrees, 13.8 degrees, 18.0 degrees and 21.7 degrees; further comprising diffraction peaks at 8.8 °, 9.1 °, 13.6 °, and 23.9 ° 2 θ; further comprising diffraction peaks at 18.5 °, 25.0 °, 26.2 ° and 27.5 ° 2 θ.
Characteristic diffraction peaks of X-rays expressed in terms of 2 theta angle and interplanar spacing d using Cu-K.alpha.radiation are shown in Table 2.
TABLE 2
Figure BDA0003005435270000112
The compound (S) -1'- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) -3-fluoropyrazin-2-yl) -1, 3-dihydrospiro [ indene-2, 4' -piperidine ] -1-amine hydrochloride crystal form B has a DSC pattern basically shown in figure 4; the TGA profile is substantially as shown in figure 5.
In a preferred embodiment of the present invention, the compound (S) -1'- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) -3-fluoropyrazin-2-yl) -1, 3-dihydrospiro [ indene-2, 4' -piperidine ] -1-amine is in the form of its ethanesulfonate in crystalline form a, the number of ethanesulfonic acids is 1 or 2, preferably the salt formation ratio of the compound of formula (III) to ethanesulfonic acids is 1: 2. spectra analysis by ethanesulfonate nuclear magnetic calibration content (fig. 17) results: a group of triplet peaks at delta 1.07-1.11ppm, which are attributed to ethanesulfonic acid methyl, have a single proton absorption peak area of 4.820/3-1.607, an average single proton absorption peak area of the compound of formula (III) of about 0.783, and an ethanesulfonic acid salt formation ratio of 1.607/0.783-2, and are calculated by calculating the compound of formula (III) and ethanesulfonic acid of 1: 2. It can be seen that the salt formation ratio of the free base of the compound of formula (III) to ethanesulfonic acid may be 1: 2.
Wherein the X-ray powder diffraction pattern has diffraction peaks at 2 theta of 8.3 degrees, 11.6 degrees and 18.3 degrees; further comprising diffraction peaks at 13.4 °, 15.3 °, 18.5 °, 20.4 °, 21.0 °, and 23.4 ° 2 θ; further comprising diffraction peaks at 10.7 °, 12.6 °, 14.1 °, 21.9 °, 22.5 °, and 25.3 ° 2 θ. Preferably, the XRPD pattern is substantially as shown in figure 6.
The compound (S) -1'- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) -3-fluoropyrazin-2-yl) -1, 3-dihydrospiro [ indene-2, 4' -piperidine ] -1-amine of the invention has ethane sulfonate crystal form A, and a DSC spectrum which is basically as shown in figure 7; the TGA profile is substantially as shown in figure 8.
In a preferred embodiment of the present invention, the compound (S) -1'- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) -3-fluoropyrazin-2-yl) -1, 3-dihydrospiro [ indene-2, 4' -piperidine ] -1-amine is in the form of a besylate salt in crystalline form a, the number of besylate acids being 1 or 2.
Wherein the X-ray powder diffraction pattern has diffraction peaks at 2 theta of 12.8 degrees, 15.3 degrees, 21.4 degrees and 25.8 degrees; further comprising diffraction peaks at 7.4 °, 11.1 ° and 22.3 ° 2 θ; further comprising diffraction peaks at 14.8 °, 16.2 °, 18.1 °, 19.8 °, 20.9 °, 23.8 °, and 24.7 ° 2 θ; further comprising diffraction peaks at 15.9 °, 16.5 °, 19.0 °, 22.8 °, 27.3 °, and 28.3 ° 2 θ.
Characteristic diffraction peaks of X-rays expressed in terms of 2. theta. angle and interplanar spacing d using Cu-K.alpha.radiation are shown in Table 4.
TABLE 3
Figure BDA0003005435270000121
Figure BDA0003005435270000131
The DSC spectrum of the benzene sulfonate crystal form A of the compound (S) -1'- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) -3-fluoropyrazin-2-yl) -1, 3-dihydrospiro [ indene-2, 4' -piperidine ] -1-amine shown as the general formula (I) is basically shown as figure 9; the TGA profile is substantially as shown in figure 10.
In a preferred embodiment of the present invention, the compound (S) -1'- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) -3-fluoropyrazin-2-yl) -1, 3-dihydrospiro [ indene-2, 4' -piperidin ] -1-amine is in the form of the fumarate salt in crystalline form a, the number of fumaric acids being 1 or 2.
Wherein the X-ray powder diffraction pattern has diffraction peaks at 8.6 DEG and 16.9 DEG 2 theta; further comprising diffraction peaks at 11.5 °, 13.0 ° and 22.1 ° 2 θ; further comprising diffraction peaks at 4.1 °, 18.6 °, 23.2 °, 23.9 °, and 26.8 ° 2 θ; further included are diffraction peaks at 19.5 °, 23.5 ° and 26.0 ° 2 θ.
Characteristic diffraction peaks of X-rays expressed in terms of 2. theta. angle and interplanar spacing d using Cu-K.alpha.radiation are shown in Table 5.
TABLE 4
Figure BDA0003005435270000132
The fumarate salt of the compound (S) -1'- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) -3-fluoropyrazin-2-yl) -1, 3-dihydrospiro [ indene-2, 4' -piperidine ] -1-amine of the invention is in a crystal form A, and the DSC spectrum of the fumarate salt is basically as shown in a figure 11; the TGA profile is substantially as shown in figure 12.
In a preferred embodiment of the present invention, the compound (S) -1'- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) -3-fluoropyrazin-2-yl) -1, 3-dihydrospiro [ indene-2, 4' -piperidine ] -1-amine is in the form of isethionate salt in crystalline form a, the number of isethionic acids being 1 or 2.
Wherein the X-ray powder diffraction pattern has diffraction peaks at 2 theta of 12.3 degrees, 16.5 degrees and 24.8 degrees; further comprising diffraction peaks at 15.0 °, 18.8 °, 26.1 ° and 27.5 ° 2 θ; further comprising diffraction peaks at 14.0 °, 16.2 ° and 18.7 ° 2 θ.
Characteristic diffraction peaks of X-rays expressed in terms of 2. theta. angle and interplanar spacing d using Cu-K.alpha.radiation are shown in Table 6.
TABLE 5
Figure BDA0003005435270000141
The compound (S) -1'- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) -3-fluoropyrazin-2-yl) -1, 3-dihydrospiro [ indene-2, 4' -piperidine ] -1-amine of the invention has a crystal form A of isethionate salt, and a DSC spectrum which is basically as shown in figure 13; the TGA profile is substantially as shown in figure 14.
In a preferred embodiment of the present invention, the compound (S) -1'- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) -3-fluoropyrazin-2-yl) -1, 3-dihydrospiro [ indene-2, 4' -piperidine ] -1-amine is in the form of the oxalate salt in form a, the number of oxalic acid being 1 or 2.
Wherein the X-ray powder diffraction pattern has diffraction peaks at 14.3 DEG, 17.8 DEG and 23.3 DEG 2 theta; further comprising diffraction peaks at 6.6 °, 11.9 °, 20.2 ° and 26.1 ° 2 θ; further comprising diffraction peaks at 8.7 °, 15.7 °, 24.5 °, 27.6 °, and 28.9 ° 2 θ.
Characteristic diffraction peaks of X-rays expressed in terms of 2. theta. angle and interplanar spacing d using Cu-K.alpha.radiation are shown in Table 7.
TABLE 6
Figure BDA0003005435270000142
Figure BDA0003005435270000151
The DSC pattern of the oxalate salt crystal form A of the compound (S) -1'- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) -3-fluoropyrazin-2-yl) -1, 3-dihydrospiro [ indene-2, 4' -piperidine ] -1-amine is basically shown in figure 15; the TGA profile is substantially as shown in figure 16.
In a preferred embodiment of the present invention, the compound (S) -1'- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) -3-fluoropyrazin-2-yl) -1, 3-dihydrospiro [ indene-2, 4' -piperidine ] -1-amine is in the form of the hydrobromide salt in crystalline form a, the number of the acids being 1 or 2.
Wherein the X-ray powder diffraction pattern has diffraction peaks at 13.0 DEG, 19.3 DEG and 25.2 DEG 2 theta; further comprising diffraction peaks at 7.1 °, 24.3 °, 26.6 °, and 27.1 ° 2 θ; further comprising diffraction peaks at 6.0 °, 12.7 °, 14.0 °, 19.9 °, 21.6 °, 22.0 °, 25.7 °, and 26.3 ° 2 θ; further included are diffraction peaks at 11.1 °, 12.0 °, 20.3 °, 22.5 °, 27.9 °, and 30.5 ° 2 θ.
Characteristic diffraction peaks of X-rays expressed in terms of 2. theta. angle and interplanar spacing d using Cu-K.alpha.radiation are shown in Table 8.
TABLE 7
Figure BDA0003005435270000152
Figure BDA0003005435270000161
In a preferred embodiment of the present invention, the compound (S) -1'- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) -3-fluoropyrazin-2-yl) -1, 3-dihydrospiro [ indene-2, 4' -piperidine ] -1-amine is in crystalline form a gentisate salt form, the number of gentisic acid being 1 or 2.
Wherein the X-ray powder diffraction pattern has diffraction peaks at 2 theta of 12.4 degrees, 17.6 degrees, 20.8 degrees and 22.2 degrees; further comprising diffraction peaks at 7.5 °, 8.5 °, 15.8 °, and 25.8 ° 2 θ; further comprising diffraction peaks at 5.7 °, 10.6 °, 14.8 °, 17.1 °, 18.9 °, and 24.9 ° 2 θ.
Characteristic diffraction peaks of X-rays expressed in terms of 2. theta. angle and interplanar spacing d using Cu-K.alpha.radiation are shown in Table 9.
TABLE 8
Figure BDA0003005435270000162
Figure BDA0003005435270000171
In a further preferred embodiment of the present invention, there is provided crystalline mesylate form a of the compound (S) -8- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) pyrazin-2-yl) -2-cyclopropyl-8-azaspiro [4.5] dec-2-en-1-amine.
Wherein the X-ray powder diffraction pattern of the mesylate crystal form A has a diffraction peak at 20.0 +/-0.2 degrees; or a diffraction peak at 17.8 ± 0.2 °; or a diffraction peak at 24.2 ± 0.2 °; or a diffraction peak at 22.2 ± 0.2 °; or a diffraction peak at 24.6 ± 0.2 °; or a diffraction peak at 12.5 ± 0.2 °; or a diffraction peak at 28.6 ± 0.2 °; or a diffraction peak at 18.8 ± 0.2 °; or a diffraction peak at 11.0 ± 0.2 °; or a diffraction peak at 14.2 ± 0.2 °; preferably any 2-5, or 3-6, or 3-8, or 5-8, or 6-8 of the above diffraction peaks, more preferably any 6, 7 or 8 thereof;
Preferably, the mesylate salt form a has an X-ray powder diffraction pattern comprising at least one or more diffraction peaks, preferably 2, more preferably 3, located at 2 Θ at 20.0 ± 0.2 °, 17.8 ± 0.2 °, 24.2 ± 0.2 °; optionally, the compound further comprises at least one position with 2 theta of 22.2 +/-0.2 degrees, 24.6 +/-0.2 degrees, 12.5 +/-0.2 degrees, 28.6 +/-0.2 degrees and 18.8 +/-0.2 degrees, preferably comprises 2, 3, 4 or 5 positions,
for example, the mesylate salt form a has an X-ray powder diffraction pattern with diffraction peaks at the following positions 2 θ:
at 20.0 +/-0.2 degrees and 17.8 +/-0.2 degrees,
or 17.8 +/-0.2 degrees and 24.2 +/-0.2 degrees,
or 20.0 +/-0.2 degrees and 24.2 +/-0.2 degrees,
or 20.0 +/-0.2 degrees, 17.8 +/-0.2 degrees and 24.2 +/-0.2 degrees,
or 17.8 +/-0.2 degrees, 24.2 +/-0.2 degrees and 22.2 +/-0.2 degrees,
or 20.0 +/-0.2 degrees, 17.8 +/-0.2 degrees, 24.2 +/-0.2 degrees and 22.2 +/-0.2 degrees,
or 17.8 +/-0.2 degrees, 24.2 +/-0.2 degrees, 22.2 +/-0.2 degrees and 24.6 +/-0.2 degrees,
or 20.0 +/-0.2 degrees, 17.8 +/-0.2 degrees, 24.2 +/-0.2 degrees, 22.2 +/-0.2 degrees and 24.6 +/-0.2 degrees,
or 17.8 +/-0.2 degrees, 24.2 +/-0.2 degrees, 22.2 +/-0.2 degrees, 24.6 +/-0.2 degrees and 12.5 +/-0.2 degrees,
more preferably, the X-ray powder diffraction pattern of mesylate form a optionally further comprises one or more diffraction peaks at 11.0 ± 0.2 °, 14.2 ± 0.2 °, 28.2 ± 0.2 °, 21.9 ± 0.2 °, 26.6 ± 0.2 °, 21.2 ± 0.2 °, 29.7 ± 0.2 ° 2 Θ; preferably at least any 2-3, or 4-5, or 6-7 thereof; further preferably, any 2, 3, 4, 5, 6, 7,
For example, the mesylate salt form a has an X-ray powder diffraction pattern with diffraction peaks at the following positions 2 θ:
11.0 +/-0.2 degrees, 14.2 +/-0.2 degrees, 28.2 +/-0.2 degrees, 21.9 +/-0.2 degrees, 26.6 +/-0.2 degrees and 21.2 +/-0.2 degrees,
or 14.2 +/-0.2 degrees, 28.2 +/-0.2 degrees, 21.9 +/-0.2 degrees, 26.6 +/-0.2 degrees, 21.2 +/-0.2 degrees and 29.7 +/-0.2 degrees,
or 11.0 +/-0.2 degrees, 14.2 +/-0.2 degrees, 21.9 +/-0.2 degrees, 26.6 +/-0.2 degrees, 21.2 +/-0.2 degrees and 29.7 +/-0.2 degrees,
further preferably, the X-ray powder diffraction pattern of mesylate form a comprises diffraction peaks at one or more of 20.0 ± 0.2 °, 17.8 ± 0.2 °, 24.2 ± 0.2 °, 22.2 ± 0.2 °, 24.6 ± 0.2 °, 12.5 ± 0.2 °, 28.6 ± 0.2 °, 18.8 ± 0.2 °, 11.0 ± 0.2 °, 14.2 ± 0.2 °, 28.2 ± 0.2 °, 21.9 ± 0.2 °, 26.6 ± 0.2 °, 21.2 ± 0.2 °, and 29.7 ± 0.2 ° of 2 Θ; preferably, the compound contains diffraction peaks at 4, 5, 6, 8 and 10,
for example, the mesylate salt form a has an X-ray powder diffraction pattern with diffraction peaks at the following positions 2 θ:
at 20.0 +/-0.2 degrees, 17.8 +/-0.2 degrees, 24.2 +/-0.2 degrees and 29.7 +/-0.2 degrees,
or 17.8 +/-0.2 degrees, 24.2 +/-0.2 degrees, 22.2 +/-0.2 degrees and 21.2 +/-0.2 degrees,
or 24.2 +/-0.2 degrees, 22.2 +/-0.2 degrees, 24.6 +/-0.2 degrees, 12.5 +/-0.2 degrees and 28.6 +/-0.2 degrees,
Or 22.2 +/-0.2 degrees, 24.6 +/-0.2 degrees, 12.5 +/-0.2 degrees, 28.6 +/-0.2 degrees and 18.8 +/-0.2 degrees,
or 20.0 +/-0.2 degrees, 17.8 +/-0.2 degrees, 24.2 +/-0.2 degrees, 24.6 +/-0.2 degrees, 12.5 +/-0.2 degrees and 28.6 +/-0.2 degrees,
or 17.8 +/-0.2 degrees, 24.2 +/-0.2 degrees, 22.2 +/-0.2 degrees, 24.6 +/-0.2 degrees, 28.6 +/-0.2 degrees and 18.8 +/-0.2 degrees,
or 24.2 +/-0.2 degrees, 22.2 +/-0.2 degrees, 24.6 +/-0.2 degrees, 12.5 +/-0.2 degrees, 28.6 +/-0.2 degrees, 18.8 +/-0.2 degrees, 11.0 +/-0.2 degrees and 14.2 +/-0.2 degrees,
or 22.2 +/-0.2 degrees, 24.6 +/-0.2 degrees, 12.5 +/-0.2 degrees, 28.6 +/-0.2 degrees, 18.8 +/-0.2 degrees, 11.0 +/-0.2 degrees, 14.2 +/-0.2 degrees and 28.2 +/-0.2 degrees,
or 20.0 +/-0.2 degrees, 17.8 +/-0.2 degrees, 24.2 +/-0.2 degrees, 22.2 +/-0.2 degrees, 24.6 +/-0.2 degrees, 12.5 +/-0.2 degrees, 28.6 +/-0.2 degrees, 18.8 +/-0.2 degrees, 11.0 +/-0.2 degrees and 14.2 +/-0.2 degrees,
or 17.8 +/-0.2 degrees, 24.2 +/-0.2 degrees, 22.2 +/-0.2 degrees, 24.6 +/-0.2 degrees, 12.5 +/-0.2 degrees, 28.6 +/-0.2 degrees, 18.8 +/-0.2 degrees, 11.0 +/-0.2 degrees, 14.2 +/-0.2 degrees and 28.2 +/-0.2 degrees.
Most preferably, using Cu-K.alpha.radiation, the characteristic diffraction peaks of X-rays expressed in terms of 2 theta angle and interplanar spacing d are shown in Table 9.
TABLE 9
Figure BDA0003005435270000191
The mesylate crystal form A of the compound (S) -8- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) pyrazin-2-yl) -2-cyclopropyl-8-azaspiro [4.5] dec-2-en-1-amine has an X-ray powder diffraction pattern basically as shown in figure 25; the DSC pattern is basically shown in figure 26.
In a further preferred embodiment of the present invention, the compound (S) -8- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) pyrazin-2-yl) -2-cyclopropyl-8-azaspiro [4.5] dec-2-en-1-amine isethionate crystalline form a is provided.
Wherein the isethionate salt crystalline form a of the compound (S) -8- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) pyrazin-2-yl) -2-cyclopropyl-8-azaspiro [4.5] dec-2-en-1-amine has a diffraction peak at 20.1 ± 0.2 ° in its X-ray powder diffraction pattern; or a diffraction peak at 18.7 ± 0.2 °; or a diffraction peak at 23.4 ± 0.2 °; or a diffraction peak at 18.1 ± 0.2 °; or a diffraction peak at 20.9 ± 0.2 °; or a diffraction peak at 23.2 ± 0.2 °; or a diffraction peak at 12.6 ± 0.2 °; or a diffraction peak at 13.3 ± 0.2 °; or a diffraction peak at 11.6 ± 0.2 °; or a diffraction peak at 10.4 ± 0.2 °; preferably any 2-5, or 3-6, or 3-8, or 5-8, or 6-8 of the above diffraction peaks, more preferably any 6, 7 or 8 thereof;
preferably, the compound (S) -8- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) pyrazin-2-yl) -2-cyclopropyl-8-azaspiro [4.5] dec-2-en-1-amine isethionate form a has an X-ray powder diffraction pattern comprising at least one or more diffraction peaks, preferably 2, more preferably 3, located at 2 Θ of 20.1 ± 0.2 °, 18.7 ± 0.2 °, 23.4 ± 0.2 °; optionally, the compound further comprises at least one position with 2 theta of 18.1 +/-0.2 degrees, 20.9 +/-0.2 degrees, 23.2 +/-0.2 degrees, 12.6 +/-0.2 degrees and 13.3 +/-0.2 degrees, preferably comprises 2, 3, 4 or 5 positions,
For example, the isethionate salt form a has an X-ray powder diffraction pattern with diffraction peaks at the following positions 2 θ:
at 20.1 +/-0.2 degrees and 18.7 +/-0.2 degrees,
or 18.7 +/-0.2 degrees and 23.4 +/-0.2 degrees,
or 20.1 +/-0.2 degrees, 18.7 +/-0.2 degrees and 23.4 +/-0.2 degrees,
or at 18.7 +/-0.2 degrees, 23.4 +/-0.2 degrees and 18.1 +/-0.2 degrees,
or 20.1 +/-0.2 degrees, 18.7 +/-0.2 degrees, 23.4 +/-0.2 degrees and 18.1 +/-0.2 degrees,
or at 18.7 +/-0.2 degrees, 23.4 +/-0.2 degrees, 18.1 +/-0.2 degrees and 20.9 +/-0.2 degrees,
or 20.1 +/-0.2 degrees, 18.7 +/-0.2 degrees, 23.4 +/-0.2 degrees, 18.1 +/-0.2 degrees and 20.9 +/-0.2 degrees,
or at 18.7 + -0.2 deg., 23.4 + -0.2 deg., 18.1 + -0.2 deg., 20.9 + -0.2 deg. and 23.2 + -0.2 deg.,
more preferably, the compound (S) -8- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) pyrazin-2-yl) -2-cyclopropyl-8-azaspiro [4.5] dec-2-en-1-amine isethionate crystalline form a has an X-ray powder diffraction pattern optionally further comprising one or more diffraction peaks at 2 Θ of 11.6 ± 0.2 °, 10.4 ± 0.2 °, 16.8 ± 0.2 °, 19.8 ± 0.2 °, 24.5 ± 0.2 °, 28.0 ± 0.2 °, 24.9 ± 0.2 °; preferably at least any 2-3, or 4-5, or 6-7 thereof; further preferably, any 2, 3, 4, 5, 6, 7,
For example, the isethionate salt form a has an X-ray powder diffraction pattern with diffraction peaks at the following positions 2 θ:
11.6 +/-0.2 degrees, 10.4 +/-0.2 degrees, 16.8 +/-0.2 degrees, 19.8 +/-0.2 degrees, 24.5 +/-0.2 degrees and 28.0 +/-0.2 degrees,
or 10.4 +/-0.2 degrees, 16.8 +/-0.2 degrees, 19.8 +/-0.2 degrees, 24.5 +/-0.2 degrees, 28.0 +/-0.2 degrees and 24.9 +/-0.2 degrees,
or 11.6 +/-0.2 degrees, 10.4 +/-0.2 degrees, 19.8 +/-0.2 degrees, 24.5 +/-0.2 degrees, 28.0 +/-0.2 degrees and 24.9 +/-0.2 degrees,
further preferably, the compound (S) -8- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) pyrazin-2-yl) -2-cyclopropyl-8-azaspiro [4.5] dec-2-ene-1-amine isethionate form a has an X-ray powder diffraction pattern comprising diffraction peaks at one or more of the positions of 20.1 ± 0.2 °, 18.7 ± 0.2 °, 23.4 ± 0.2 °, 18.1 ± 0.2 °, 20.9 ± 0.2 °, 23.2 ± 0.2 °, 12.6 ± 0.2 °, 13.3 ± 0.2 °, 11.6 ± 0.2 °, 10.4 ± 0.2 °, 16.8 ± 0.2 °, 19.8 ± 0.2 °, 24.5 ± 0.2 °, 28.0 ± 0.2 °, 24.9 ± 0.2 ° 2 Θ; preferably, the compound contains diffraction peaks at 4, 5, 6, 8 and 10,
for example, the isethionate salt form a has an X-ray powder diffraction pattern with diffraction peaks at the following positions 2 θ:
20.1 +/-0.2 degrees, 18.1 +/-0.2 degrees, 20.9 +/-0.2 degrees and 23.2 +/-0.2 degrees,
or 18.7 +/-0.2 degrees, 18.1 +/-0.2 degrees, 20.9 +/-0.2 degrees and 23.2 +/-0.2 degrees,
or 23.4 +/-0.2 degrees, 18.1 +/-0.2 degrees, 20.9 +/-0.2 degrees, 23.2 +/-0.2 degrees and 12.6 +/-0.2 degrees,
or 18.1 +/-0.2 degrees, 20.9 +/-0.2 degrees, 23.2 +/-0.2 degrees, 12.6 +/-0.2 degrees and 13.3 +/-0.2 degrees,
or 20.1 +/-0.2 degrees, 18.7 +/-0.2 degrees, 23.4 +/-0.2 degrees, 18.1 +/-0.2 degrees, 20.9 +/-0.2 degrees and 23.2 +/-0.2 degrees,
or 18.7 +/-0.2 degrees, 23.4 +/-0.2 degrees, 18.1 +/-0.2 degrees, 20.9 +/-0.2 degrees, 23.2 +/-0.2 degrees and 12.6 +/-0.2 degrees,
or 23.4 +/-0.2 degrees, 18.1 +/-0.2 degrees, 20.9 +/-0.2 degrees, 23.2 +/-0.2 degrees, 12.6 +/-0.2 degrees, 13.3 +/-0.2 degrees, 11.6 +/-0.2 degrees and 10.4 +/-0.2 degrees,
or 18.1 +/-0.2 degrees, 20.9 +/-0.2 degrees, 23.2 +/-0.2 degrees, 12.6 +/-0.2 degrees, 13.3 +/-0.2 degrees, 11.6 +/-0.2 degrees, 10.4 +/-0.2 degrees and 16.8 +/-0.2 degrees,
or 20.1 +/-0.2 degrees, 18.7 +/-0.2 degrees, 23.4 +/-0.2 degrees, 18.1 +/-0.2 degrees, 20.9 +/-0.2 degrees, 23.2 +/-0.2 degrees, 12.6 +/-0.2 degrees, 13.3 +/-0.2 degrees, 11.6 +/-0.2 degrees and 10.4 +/-0.2 degrees,
or 18.7 +/-0.2 degrees, 23.4 +/-0.2 degrees, 18.1 +/-0.2 degrees, 20.9 +/-0.2 degrees, 23.2 +/-0.2 degrees, 12.6 +/-0.2 degrees, 13.3 +/-0.2 degrees, 11.6 +/-0.2 degrees, 10.4 +/-0.2 degrees and 16.8 +/-0.2 degrees.
The isethionate salt crystal form a of the compound (S) -8- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) pyrazin-2-yl) -2-cyclopropyl-8-azaspiro [4.5] dec-2-en-1-amine of the present invention has X-ray characteristic diffraction peaks expressed in terms of 2 theta angles and interplanar spacings d values using Cu-ka radiation as shown in table 10.
Watch 10
Figure BDA0003005435270000211
Figure BDA0003005435270000221
The isethionate crystal form A of the compound (S) -8- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) pyrazin-2-yl) -2-cyclopropyl-8-azaspiro [4.5] dec-2-en-1-amine of the present invention has an X-ray powder diffraction pattern substantially as shown in FIG. 27; the DSC pattern is basically shown in figure 28.
In a preferred embodiment of the present invention, the compound (S) -8- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) pyrazin-2-yl) -2-cyclopropyl-8-azaspiro [4.5] dec-2-en-1-amine is in the form of fumarate salt form a, the number of fumaric acids being 1.
In a further preferred embodiment of the present invention, there is provided crystalline fumarate salt form a of the compound (S) -8- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) pyrazin-2-yl) -2-cyclopropyl-8-azaspiro [4.5] dec-2-en-1-amine.
Wherein the fumarate salt of the compound (S) -8- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) pyrazin-2-yl) -2-cyclopropyl-8-azaspiro [4.5] dec-2-en-1-amine form a has an X-ray powder diffraction pattern with a diffraction peak at 18.7 ± 0.2 °; or a diffraction peak at 23.1 ± 0.2 °; or a diffraction peak at 20.1 ± 0.2 °; or a diffraction peak at 11.3 ± 0.2 °; or a diffraction peak at 25.0 ± 0.2 °; or a diffraction peak at 12.6 ± 0.2 °; or a diffraction peak at 22.0 ± 0.2 °; or a diffraction peak at 18.2 ± 0.2 °; or a diffraction peak at 27.8 ± 0.2 °; or a diffraction peak at 14.6 ± 0.2 °; preferably any 2-5, or 3-6, or 3-8, or 5-8, or 6-8 of the above diffraction peaks, more preferably any 6, 7 or 8 thereof;
Preferably, the compound (S) -8- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) pyrazin-2-yl) -2-cyclopropyl-8-azaspiro [4.5] dec-2-en-1-amine fumarate salt form a has an X-ray powder diffraction pattern comprising at least one or more diffraction peaks, preferably 2, more preferably 3, located at 2 Θ of 18.7 ± 0.2 °, 23.1 ± 0.2 °, 20.1 ± 0.2 °; optionally, the compound further comprises at least one position with 2 theta of 11.3 +/-0.2 degrees, 25.0 +/-0.2 degrees, 12.6 +/-0.2 degrees, 22.0 +/-0.2 degrees and 18.2 +/-0.2 degrees, preferably comprises 2, 3, 4 or 5 positions,
for example, the fumarate salt form a has an X-ray powder diffraction pattern with diffraction peaks at the following positions 2 θ:
at 18.7 +/-0.2 degrees and 23.1 +/-0.2 degrees,
or at 23.1 + -0.2 deg. and 20.1 + -0.2 deg.,
or at 18.7 +/-0.2 degrees, 23.1 +/-0.2 degrees and 20.1 +/-0.2 degrees,
or at 23.1 +/-0.2 degrees, 20.1 +/-0.2 degrees and 11.3 +/-0.2 degrees,
or at 18.7 + -0.2 deg., 23.1 + -0.2 deg., 20.1 + -0.2 deg. and 11.3 + -0.2 deg.,
or at 23.1 + -0.2 deg., 20.1 + -0.2 deg., 11.3 + -0.2 deg. and 25.0 + -0.2 deg.,
or at 18.7 + -0.2 deg., 23.1 + -0.2 deg., 20.1 + -0.2 deg., 11.3 + -0.2 deg. and 25.0 + -0.2 deg.,
or at 23.1 + -0.2 deg., 20.1 + -0.2 deg., 11.3 + -0.2 deg., 25.0 + -0.2 deg. and 12.6 + -0.2 deg.,
More preferably, the compound (S) -8- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) pyrazin-2-yl) -2-cyclopropyl-8-azaspiro [4.5] dec-2-en-1-amine fumarate crystalline form a optionally further comprises one or more diffraction peaks at 2 Θ of 27.8 ± 0.2 °, 14.6 ± 0.2 °, 5.1 ± 0.2 °, 15.4 ± 0.2 °, 28.8 ± 0.2 °, 13.9 ± 0.2 °, 20.5 ± 0.2 °; preferably at least any 2-3, or 4-5, or 6-7 thereof; further preferably, any 2, 3, 4, 5, 6, 7,
for example, the fumarate salt form a has an X-ray powder diffraction pattern with diffraction peaks at the following positions 2 θ:
27.8 +/-0.2 degrees, 14.6 +/-0.2 degrees, 5.1 +/-0.2 degrees, 15.4 +/-0.2 degrees, 28.8 +/-0.2 degrees and 13.9 +/-0.2 degrees,
or 14.6 +/-0.2 degrees, 5.1 +/-0.2 degrees, 15.4 +/-0.2 degrees, 28.8 +/-0.2 degrees, 13.9 +/-0.2 degrees and 20.5 +/-0.2 degrees,
or 27.8 +/-0.2 degrees, 14.6 +/-0.2 degrees, 15.4 +/-0.2 degrees, 28.8 +/-0.2 degrees, 13.9 +/-0.2 degrees and 20.5 +/-0.2 degrees,
further preferably, the compound (S) -8- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) pyrazin-2-yl) -2-cyclopropyl-8-azaspiro [4.5] dec-2-ene-1-amine fumarate crystalline form a has an X-ray powder diffraction pattern comprising diffraction peaks at one or more of 2 Θ at 18.7 ± 0.2 °, 23.1 ± 0.2 °, 20.1 ± 0.2 °, 11.3 ± 0.2 °, 25.0 ± 0.2 °, 12.6 ± 0.2 °, 22.0 ± 0.2 °, 18.2 ± 0.2 °, 27.8 ± 0.2 °, 14.6 ± 0.2 °, 5.1 ± 0.2 °, 15.4 ± 0.2 °, 28.8 ± 0.2 °, 13.9 ± 0.2 °, 20.5 ± 0.2 °; preferably, the compound contains diffraction peaks at 4, 5, 6, 8 and 10,
For example, the fumarate salt form a has an X-ray powder diffraction pattern with diffraction peaks at the following positions 2 θ:
at 18.7 +/-0.2 degrees, 20.1 +/-0.2 degrees, 11.3 +/-0.2 degrees and 25.0 +/-0.2 degrees,
or at 23.1 +/-0.2 degrees, 11.3 +/-0.2 degrees, 25.0 +/-0.2 degrees and 12.6 +/-0.2 degrees,
or 20.1 +/-0.2 degrees, 11.3 +/-0.2 degrees, 25.0 +/-0.2 degrees, 12.6 +/-0.2 degrees and 22.0 +/-0.2 degrees,
or 11.3 +/-0.2 degrees, 25.0 +/-0.2 degrees, 12.6 +/-0.2 degrees, 22.0 +/-0.2 degrees and 18.2 +/-0.2 degrees,
or 18.7 +/-0.2 degrees, 23.1 +/-0.2 degrees, 20.1 +/-0.2 degrees, 11.3 +/-0.2 degrees, 25.0 +/-0.2 degrees and 12.6 +/-0.2 degrees,
or 23.1 +/-0.2 degrees, 20.1 +/-0.2 degrees, 11.3 +/-0.2 degrees, 25.0 +/-0.2 degrees, 12.6 +/-0.2 degrees and 22.0 +/-0.2 degrees,
or 20.1 +/-0.2 degrees, 11.3 +/-0.2 degrees, 25.0 +/-0.2 degrees, 12.6 +/-0.2 degrees, 22.0 +/-0.2 degrees, 18.2 +/-0.2 degrees, 27.8 +/-0.2 degrees and 14.6 +/-0.2 degrees,
or 11.3 +/-0.2 degrees, 25.0 +/-0.2 degrees, 12.6 +/-0.2 degrees, 22.0 +/-0.2 degrees, 18.2 +/-0.2 degrees, 27.8 +/-0.2 degrees, 14.6 +/-0.2 degrees and 5.1 +/-0.2 degrees,
or 18.7 +/-0.2 degrees, 23.1 +/-0.2 degrees, 20.1 +/-0.2 degrees, 11.3 +/-0.2 degrees, 25.0 +/-0.2 degrees, 12.6 +/-0.2 degrees, 22.0 +/-0.2 degrees, 18.2 +/-0.2 degrees, 27.8 +/-0.2 degrees and 14.6 +/-0.2 degrees,
or 23.1 +/-0.2 degrees, 20.1 +/-0.2 degrees, 11.3 +/-0.2 degrees, 25.0 +/-0.2 degrees, 12.6 +/-0.2 degrees, 22.0 +/-0.2 degrees, 18.2 +/-0.2 degrees, 27.8 +/-0.2 degrees, 14.6 +/-0.2 degrees and 5.1 +/-0.2 degrees.
The fumarate salt of the compound (S) -8- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) pyrazin-2-yl) -2-cyclopropyl-8-azaspiro [4.5] dec-2-en-1-amine of the present invention in crystalline form a, characteristic diffraction peaks of X-rays expressed in terms of 2 θ angles and interplanar spacings d values using Cu — ka radiation are shown in table 11.
TABLE 11
Figure BDA0003005435270000241
Figure BDA0003005435270000251
The fumarate crystal form A of the compound (S) -8- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) pyrazin-2-yl) -2-cyclopropyl-8-azaspiro [4.5] dec-2-en-1-amine has an X-ray powder diffraction pattern basically shown in a figure 29; the DSC pattern is basically shown in figure 30.
In a preferred embodiment of the present invention, the compound (S) -8- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) pyrazin-2-yl) -2-cyclopropyl-8-azaspiro [4.5] dec-2-en-1-amine is in crystalline form p-toluenesulfonate form a, the number of p-toluenesulphonic acids is 1.
In a further preferred embodiment of the present invention, there is provided crystalline form a of the p-toluenesulfonate salt of the compound (S) -8- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) pyrazin-2-yl) -2-cyclopropyl-8-azaspiro [4.5] dec-2-en-1-amine.
Wherein the X-ray powder diffraction pattern of the p-toluenesulfonic acid salt crystal form A of the compound (S) -8- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) pyrazin-2-yl) -2-cyclopropyl-8-azaspiro [4.5] dec-2-en-1-amine has a diffraction peak at 23.2 +/-0.2 degrees; or a diffraction peak at 11.2 ± 0.2 °; or a diffraction peak at 19.4 ± 0.2 °; or a diffraction peak at 16.6 ± 0.2 °; or a diffraction peak at 21.1 ± 0.2 °; or a diffraction peak at 17.3 ± 0.2 °; or a diffraction peak at 16.8 ± 0.2 °; or a diffraction peak at 4.3 ± 0.2 °; or a diffraction peak at 18.2 ± 0.2 °; or a diffraction peak at 22.0 ± 0.2 °; preferably any 2-5, or 3-6, or 3-8, or 5-8, or 6-8 of the above diffraction peaks, more preferably any 6, 7 or 8 thereof;
Preferably, the compound (S) -8- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) pyrazin-2-yl) -2-cyclopropyl-8-azaspiro [4.5] dec-2-en-1-amine p-toluenesulfonate form a has an X-ray powder diffraction pattern comprising at least one or more diffraction peaks, preferably 2, more preferably 3, located at 2 θ in the range of 23.2 ± 0.2 °, 11.2 ± 0.2 °, 19.4 ± 0.2 °; optionally, the compound further comprises at least one position with 2 theta of 16.6 +/-0.2 degrees, 21.1 +/-0.2 degrees, 17.3 +/-0.2 degrees, 16.8 +/-0.2 degrees and 4.3 +/-0.2 degrees, preferably comprises 2, 3, 4 or 5 positions,
for example, the p-toluenesulfonate form a has an X-ray powder diffraction pattern with diffraction peaks at the following positions 2 θ:
at 23.2 +/-0.2 degrees and 11.2 +/-0.2 degrees,
or 11.2 +/-0.2 degrees and 19.4 +/-0.2 degrees,
or at 23.2 + -0.2 deg., 11.2 + -0.2 deg. and 19.4 + -0.2 deg.,
or 11.2 +/-0.2 degrees, 19.4 +/-0.2 degrees and 16.6 +/-0.2 degrees,
or at 23.2 + -0.2 deg., 11.2 + -0.2 deg., 19.4 + -0.2 deg. and 16.6 + -0.2 deg.,
or 11.2 +/-0.2 degrees, 19.4 +/-0.2 degrees, 16.6 +/-0.2 degrees and 21.1 +/-0.2 degrees,
or at 23.2 + -0.2 deg., 11.2 + -0.2 deg., 19.4 + -0.2 deg., 16.6 + -0.2 deg. and 21.1 + -0.2 deg.,
or 11.2 +/-0.2 degrees, 19.4 +/-0.2 degrees, 16.6 +/-0.2 degrees, 21.1 +/-0.2 degrees and 17.3 +/-0.2 degrees,
More preferably, the compound (S) -8- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) pyrazin-2-yl) -2-cyclopropyl-8-azaspiro [4.5] dec-2-en-1-amine p-toluenesulfonate form a optionally further comprises one or more diffraction peaks at 2 Θ of 18.2 ± 0.2 °, 22.0 ± 0.2 °, 29.2 ± 0.2 °, 20.7 ± 0.2 °, 9.6 ± 0.2 °, 26.9 ± 0.2 °, 13.2 ± 0.2 °; preferably at least any 2-3, or 4-5, or 6-7 thereof; further preferably, any 2, 3, 4, 5, 6, 7,
for example, the p-toluenesulfonate form a has an X-ray powder diffraction pattern with diffraction peaks at the following positions 2 θ:
18.2 +/-0.2 degrees, 22.0 +/-0.2 degrees, 29.2 +/-0.2 degrees, 20.7 +/-0.2 degrees, 9.6 +/-0.2 degrees and 26.9 +/-0.2 degrees,
or 22.0 +/-0.2 degrees, 29.2 +/-0.2 degrees, 20.7 +/-0.2 degrees, 9.6 +/-0.2 degrees, 26.9 +/-0.2 degrees and 13.2 +/-0.2 degrees,
or 18.2 +/-0.2 degrees, 22.0 +/-0.2 degrees, 20.7 +/-0.2 degrees, 9.6 +/-0.2 degrees, 26.9 +/-0.2 degrees and 13.2 +/-0.2 degrees,
further preferably, the compound (S) -8- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) pyrazin-2-yl) -2-cyclopropyl-8-azaspiro [4.5] dec-2-ene-1-amine p-toluenesulfonate form a has an X-ray powder diffraction pattern comprising diffraction peaks at one or more of 2 θ of 23.2 ± 0.2 °, 11.2 ± 0.2 °, 19.4 ± 0.2 °, 16.6 ± 0.2 °, 21.1 ± 0.2 °, 17.3 ± 0.2 °, 16.8 ± 0.2 °, 4.3 ± 0.2 °, 18.2 ± 0.2 °, 22.0 ± 0.2 °, 29.2 ± 0.2 °, 20.7 ± 0.2 °, 9.6 ± 0.2 °, 26.9 ± 0.2 °, 13.2 ± 0.2 °; preferably, the compound contains diffraction peaks at 4, 5, 6, 8 and 10,
For example, the p-toluenesulfonate form a has an X-ray powder diffraction pattern with diffraction peaks at the following positions 2 θ:
23.2 +/-0.2 degrees, 19.4 +/-0.2 degrees, 16.6 +/-0.2 degrees and 21.1 +/-0.2 degrees,
or 11.2 +/-0.2 degrees, 16.6 +/-0.2 degrees, 21.1 +/-0.2 degrees and 17.3 +/-0.2 degrees,
or 19.4 +/-0.2 degrees, 16.6 +/-0.2 degrees, 21.1 +/-0.2 degrees, 17.3 +/-0.2 degrees and 16.8 +/-0.2 degrees,
or 16.6 +/-0.2 degrees, 21.1 +/-0.2 degrees, 17.3 +/-0.2 degrees, 16.8 +/-0.2 degrees and 4.3 +/-0.2 degrees,
or 23.2 +/-0.2 degrees, 11.2 +/-0.2 degrees, 19.4 +/-0.2 degrees, 16.6 +/-0.2 degrees, 21.1 +/-0.2 degrees and 17.3 +/-0.2 degrees,
or 11.2 +/-0.2 degrees, 19.4 +/-0.2 degrees, 16.6 +/-0.2 degrees, 21.1 +/-0.2 degrees, 17.3 +/-0.2 degrees and 16.8 +/-0.2 degrees,
or 19.4 +/-0.2 degrees, 16.6 +/-0.2 degrees, 21.1 +/-0.2 degrees, 17.3 +/-0.2 degrees, 16.8 +/-0.2 degrees, 4.3 +/-0.2 degrees, 18.2 +/-0.2 degrees and 22.0 +/-0.2 degrees,
or 16.6 +/-0.2 degrees, 21.1 +/-0.2 degrees, 17.3 +/-0.2 degrees, 16.8 +/-0.2 degrees, 4.3 +/-0.2 degrees, 18.2 +/-0.2 degrees, 22.0 +/-0.2 degrees and 29.2 +/-0.2 degrees,
or 23.2 +/-0.2 degrees, 11.2 +/-0.2 degrees, 19.4 +/-0.2 degrees, 16.6 +/-0.2 degrees, 21.1 +/-0.2 degrees, 17.3 +/-0.2 degrees, 16.8 +/-0.2 degrees, 4.3 +/-0.2 degrees, 18.2 +/-0.2 degrees and 22.0 +/-0.2 degrees,
or 11.2 +/-0.2 degrees, 19.4 +/-0.2 degrees, 16.6 +/-0.2 degrees, 21.1 +/-0.2 degrees, 17.3 +/-0.2 degrees, 16.8 +/-0.2 degrees, 4.3 +/-0.2 degrees, 18.2 +/-0.2 degrees, 22.0 +/-0.2 degrees and 29.2 +/-0.2 degrees.
The p-toluenesulfonate salt form A of the compound (S) -8- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) pyrazin-2-yl) -2-cyclopropyl-8-azaspiro [4.5] dec-2-en-1-amine of the present invention has characteristic X-ray diffraction peaks expressed in terms of 2 theta angles and interplanar spacings d values as shown in Table 12 using Cu-Ka radiation.
TABLE 12
Figure BDA0003005435270000271
The p-toluenesulfonate crystal form A of the compound (S) -8- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) pyrazin-2-yl) -2-cyclopropyl-8-azaspiro [4.5] dec-2-en-1-amine has an X-ray powder diffraction pattern basically shown in a figure 31; the DSC pattern is basically shown in figure 32.
In a further preferred embodiment of the present invention, there is provided ethane sulfonate crystalline form a of the compound (S) -8- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) pyrazin-2-yl) -2-cyclopropyl-8-azaspiro [4.5] dec-2-en-1-amine.
Wherein the compound (S) -8- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) pyrazin-2-yl) -2-cyclopropyl-8-azaspiro [4.5] dec-2-en-1-amine ethane sulfonate form a has an X-ray powder diffraction pattern with a diffraction peak at 17.3 ± 0.2 °; or a diffraction peak at 19.5 ± 0.2 °; or a diffraction peak at 21.6 ± 0.2 °; or a diffraction peak at 18.5 ± 0.2 °; or a diffraction peak at 23.9 ± 0.2 °; or a diffraction peak at 10.6 ± 0.2 °; or a diffraction peak at 22.2 ± 0.2 °; or a diffraction peak at 13.9 ± 0.2 °; or a diffraction peak at 26.1 ± 0.2 °; or a diffraction peak at 12.3 ± 0.2 °; preferably any 2-5, or 3-6, or 3-8, or 5-8, or 6-8 of the above diffraction peaks, more preferably any 6, 7 or 8 thereof;
Preferably, the compound (S) -8- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) pyrazin-2-yl) -2-cyclopropyl-8-azaspiro [4.5] dec-2-en-1-amine ethane sulfonate salt form a has an X-ray powder diffraction pattern comprising at least one or more diffraction peaks, preferably 2, more preferably 3, located at 2 Θ of 17.3 ± 0.2 °, 19.5 ± 0.2 °, 21.6 ± 0.2 °; optionally, the compound further comprises at least one position with 2 theta of 18.5 +/-0.2 degrees, 23.9 +/-0.2 degrees, 10.6 +/-0.2 degrees, 22.2 +/-0.2 degrees and 13.9 +/-0.2 degrees, preferably comprises 2, 3, 4 or 5 positions,
for example, the ethane sulfonate form a has an X-ray powder diffraction pattern with diffraction peaks at the following positions 2 θ:
17.3 +/-0.2 degrees and 19.5 +/-0.2 degrees,
or, at 19.5 + -0.2 deg. and 21.6 + -0.2 deg.,
or 17.3 +/-0.2 degrees, 19.5 +/-0.2 degrees and 21.6 +/-0.2 degrees,
or at 19.5 +/-0.2 degrees, 21.6 +/-0.2 degrees and 18.5 +/-0.2 degrees,
or 17.3 +/-0.2 degrees, 19.5 +/-0.2 degrees, 21.6 +/-0.2 degrees and 18.5 +/-0.2 degrees,
or 19.5 +/-0.2 degrees, 21.6 +/-0.2 degrees, 18.5 +/-0.2 degrees and 23.9 +/-0.2 degrees,
or 17.3 +/-0.2 degrees, 19.5 +/-0.2 degrees, 21.6 +/-0.2 degrees, 18.5 +/-0.2 degrees and 23.9 +/-0.2 degrees,
or 19.5 +/-0.2 degrees, 21.6 +/-0.2 degrees, 18.5 +/-0.2 degrees, 23.9 +/-0.2 degrees and 10.6 +/-0.2 degrees,
More preferably, the compound (S) -8- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) pyrazin-2-yl) -2-cyclopropyl-8-azaspiro [4.5] dec-2-en-1-amine ethane sulfonate crystalline form a optionally further comprises one or more diffraction peaks at 2 Θ of 26.1 ± 0.2 °, 12.3 ± 0.2 °, 20.7 ± 0.2 °, 21.3 ± 0.2 °, 19.0 ± 0.2 °, 28.1 ± 0.2 °, 13.6 ± 0.2 °; preferably at least any 2-3, or 4-5, or 6-7 thereof; further preferably, any 2, 3, 4, 5, 6, 7,
for example, the ethane sulfonate form a has an X-ray powder diffraction pattern with diffraction peaks at the following positions 2 θ:
26.1 +/-0.2 degrees, 12.3 +/-0.2 degrees, 20.7 +/-0.2 degrees, 21.3 +/-0.2 degrees, 19.0 +/-0.2 degrees and 28.1 +/-0.2 degrees,
or 12.3 +/-0.2 degrees, 20.7 +/-0.2 degrees, 21.3 +/-0.2 degrees, 19.0 +/-0.2 degrees, 28.1 +/-0.2 degrees and 13.6 +/-0.2 degrees,
or 26.1 +/-0.2 degrees, 12.3 +/-0.2 degrees, 21.3 +/-0.2 degrees, 19.0 +/-0.2 degrees, 28.1 +/-0.2 degrees and 13.6 +/-0.2 degrees,
further preferably, the compound (S) -8- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) pyrazin-2-yl) -2-cyclopropyl-8-azaspiro [4.5] dec-2-ene-1-amine ethanesulfonate crystalline form a has an X-ray powder diffraction pattern comprising diffraction peaks at one or more of 2 Θ at 17.3 ± 0.2 °, 19.5 ± 0.2 °, 21.6 ± 0.2 °, 18.5 ± 0.2 °, 23.9 ± 0.2 °, 10.6 ± 0.2 °, 22.2 ± 0.2 °, 13.9 ± 0.2 °, 26.1 ± 0.2 °, 12.3 ± 0.2 °, 20.7 ± 0.2 °, 21.3 ± 0.2 °, 19.0 ± 0.2 °, 28.1 ± 0.2 °, 13.6 ± 0.2 °; preferably, the compound contains diffraction peaks at 4, 5, 6, 8 and 10,
For example, the ethane sulfonate form a has an X-ray powder diffraction pattern with diffraction peaks at the following positions 2 θ:
17.3 +/-0.2 degrees, 21.6 +/-0.2 degrees, 18.5 +/-0.2 degrees and 23.9 +/-0.2 degrees,
or 19.5 +/-0.2 degrees, 23.9 +/-0.2 degrees, 10.6 +/-0.2 degrees and 22.2 +/-0.2 degrees,
or 21.6 +/-0.2 degrees, 23.9 +/-0.2 degrees, 10.6 +/-0.2 degrees, 22.2 +/-0.2 degrees and 13.9 +/-0.2 degrees,
or 18.5 +/-0.2 degrees, 10.6 +/-0.2 degrees, 22.2 +/-0.2 degrees, 13.9 +/-0.2 degrees and 26.1 +/-0.2 degrees,
or 17.3 +/-0.2 degrees, 19.5 +/-0.2 degrees, 21.6 +/-0.2 degrees, 18.5 +/-0.2 degrees, 23.9 +/-0.2 degrees and 13.9 +/-0.2 degrees,
or 19.5 +/-0.2 degrees, 21.6 +/-0.2 degrees, 18.5 +/-0.2 degrees, 23.9 +/-0.2 degrees, 10.6 +/-0.2 degrees and 26.1 +/-0.2 degrees,
or 21.6 +/-0.2 degrees, 18.5 +/-0.2 degrees, 23.9 +/-0.2 degrees, 10.6 +/-0.2 degrees, 22.2 +/-0.2 degrees, 13.9 +/-0.2 degrees, 26.1 +/-0.2 degrees and 12.3 +/-0.2 degrees,
or 18.5 +/-0.2 degrees, 23.9 +/-0.2 degrees, 10.6 +/-0.2 degrees, 22.2 +/-0.2 degrees, 13.9 +/-0.2 degrees, 26.1 +/-0.2 degrees, 12.3 +/-0.2 degrees and 20.7 +/-0.2 degrees,
or 17.3 +/-0.2 degrees, 19.5 +/-0.2 degrees, 21.6 +/-0.2 degrees, 18.5 +/-0.2 degrees, 23.9 +/-0.2 degrees, 10.6 +/-0.2 degrees, 22.2 +/-0.2 degrees, 13.9 +/-0.2 degrees, 26.1 +/-0.2 degrees and 12.3 +/-0.2 degrees,
or 19.5 +/-0.2 degrees, 21.6 +/-0.2 degrees, 18.5 +/-0.2 degrees, 23.9 +/-0.2 degrees, 10.6 +/-0.2 degrees, 22.2 +/-0.2 degrees, 13.9 +/-0.2 degrees, 26.1 +/-0.2 degrees, 12.3 +/-0.2 degrees and 20.7 +/-0.2 degrees.
The ethanesulfonate salt crystal form A of the compound (S) -8- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) pyrazin-2-yl) -2-cyclopropyl-8-azaspiro [4.5] dec-2-en-1-amine of the present invention has characteristic X-ray diffraction peaks expressed in terms of 2 theta angles and interplanar spacings d values as shown in Table 13 using Cu-Ka radiation.
Watch 13
Figure BDA0003005435270000301
The ethane sulfonate crystal form A of the compound (S) -8- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) pyrazin-2-yl) -2-cyclopropyl-8-azaspiro [4.5] dec-2-en-1-amine has an X-ray powder diffraction pattern basically shown in a figure 33; the DSC pattern is basically shown in figure 34.
In a preferred embodiment of the invention, the compound (S) -8- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) pyrazin-2-yl) -2-cyclopropyl-8-azaspiro [4.5] dec-2-en-1-amine is in the form of succinate crystal form a, the number of succinic acids being 0.5.
In a further preferred embodiment of the present invention, there is provided crystalline form a of the succinate salt of the compound (S) -8- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) pyrazin-2-yl) -2-cyclopropyl-8-azaspiro [4.5] dec-2-en-1-amine.
Wherein the compound (S) -8- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) pyrazin-2-yl) -2-cyclopropyl-8-azaspiro [4.5] dec-2-en-1-amine succinate in form a has an X-ray powder diffraction pattern with a diffraction peak at 22.8 ± 0.2 °; or a diffraction peak at 19.9 ± 0.2 °; or a diffraction peak at 18.3 ± 0.2 °; or a diffraction peak at 12.3 ± 0.2 °; or a diffraction peak at 21.7 ± 0.2 °; or a diffraction peak at 24.7 ± 0.2 °; or a diffraction peak at 11.0 ± 0.2 °; or a diffraction peak at 15.2 ± 0.2 °; or a diffraction peak at 27.4 ± 0.2 °; or a diffraction peak at 20.2 ± 0.2 °; preferably any 2-5, or 3-6, or 3-8, or 5-8, or 6-8 of the above diffraction peaks, more preferably any 6, 7 or 8 thereof;
Preferably, the compound (S) -8- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) pyrazin-2-yl) -2-cyclopropyl-8-azaspiro [4.5] dec-2-en-1-amine succinate in form a has an X-ray powder diffraction pattern comprising at least one or more diffraction peaks, preferably 2, more preferably 3, located at 2 Θ of 22.8 ± 0.2 °, 19.9 ± 0.2 °, 18.3 ± 0.2 °; optionally, the compound further comprises at least one position with 2 theta of 12.3 +/-0.2 degrees, 21.7 +/-0.2 degrees, 24.7 +/-0.2 degrees, 11.0 +/-0.2 degrees and 15.2 +/-0.2 degrees, preferably comprises 2, 3, 4 or 5 positions,
for example, the succinate form a has an X-ray powder diffraction pattern with diffraction peaks at the following positions 2 θ:
22.8 +/-0.2 degrees and 19.9 +/-0.2 degrees,
or, at 19.9 + -0.2 deg. and 18.3 + -0.2 deg.,
or at 22.8 +/-0.2 degrees, 19.9 +/-0.2 degrees and 18.3 +/-0.2 degrees,
or at 19.9 +/-0.2 degrees, 18.3 +/-0.2 degrees and 12.3 +/-0.2 degrees,
or 22.8 +/-0.2 degrees, 19.9 +/-0.2 degrees, 18.3 +/-0.2 degrees and 12.3 +/-0.2 degrees,
or 19.9 +/-0.2 degrees, 18.3 +/-0.2 degrees, 12.3 +/-0.2 degrees and 21.7 +/-0.2 degrees,
or 22.8 +/-0.2 degrees, 19.9 +/-0.2 degrees, 18.3 +/-0.2 degrees, 12.3 +/-0.2 degrees and 21.7 +/-0.2 degrees,
or 19.9 +/-0.2 degrees, 18.3 +/-0.2 degrees, 12.3 +/-0.2 degrees, 21.7 +/-0.2 degrees and 24.7 +/-0.2 degrees,
More preferably, the compound (S) -8- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) pyrazin-2-yl) -2-cyclopropyl-8-azaspiro [4.5] dec-2-en-1-amine succinate crystalline form a optionally further comprises one or more diffraction peaks at 2 Θ of 27.4 ± 0.2 °, 20.2 ± 0.2 °, 14.3 ± 0.2 °, 20.9 ± 0.2 °, 28.5 ± 0.2 °, 32.2 ± 0.2 °, 13.7 ± 0.2 °; preferably at least any 2-3, or 4-5, or 6-7 thereof; further preferably, any 2, 3, 4, 5, 6, 7,
for example, the succinate form a has an X-ray powder diffraction pattern with diffraction peaks at the following positions 2 θ:
27.4 +/-0.2 degrees, 20.2 +/-0.2 degrees, 14.3 +/-0.2 degrees, 20.9 +/-0.2 degrees, 28.5 +/-0.2 degrees and 32.2 +/-0.2 degrees,
or 20.2 +/-0.2 degrees, 14.3 +/-0.2 degrees, 20.9 +/-0.2 degrees, 28.5 +/-0.2 degrees, 32.2 +/-0.2 degrees and 13.7 +/-0.2 degrees,
or 27.4 +/-0.2 degrees, 20.2 +/-0.2 degrees, 20.9 +/-0.2 degrees, 28.5 +/-0.2 degrees, 32.2 +/-0.2 degrees and 13.7 +/-0.2 degrees,
further preferably, the compound (S) -8- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) pyrazin-2-yl) -2-cyclopropyl-8-azaspiro [4.5] dec-2-ene-1-amine succinate has an X-ray powder diffraction pattern comprising diffraction peaks at one or more of 22.8 ± 0.2 °, 19.9 ± 0.2 °, 18.3 ± 0.2 °, 12.3 ± 0.2 °, 21.7 ± 0.2 °, 24.7 ± 0.2 °, 11.0 ± 0.2 °, 15.2 ± 0.2 °, 27.4 ± 0.2 °, 20.2 ± 0.2 °, 14.3 ± 0.2 °, 20.9 ± 0.2 °, 28.5 ± 0.2 °, 32.2 ± 0.2 °, 13.7 ± 0.2 ° of 2 Θ; preferably, the compound contains diffraction peaks at 4, 5, 6, 8 and 10,
For example, the succinate form a has an X-ray powder diffraction pattern with diffraction peaks at the following positions 2 θ:
22.8 +/-0.2 degrees, 12.3 +/-0.2 degrees, 21.7 +/-0.2 degrees and 24.7 +/-0.2 degrees,
or 19.9 +/-0.2 degrees, 21.7 +/-0.2 degrees, 24.7 +/-0.2 degrees and 11.0 +/-0.2 degrees,
or 18.3 +/-0.2 degrees, 12.3 +/-0.2 degrees, 21.7 +/-0.2 degrees, 24.7 +/-0.2 degrees and 11.0 +/-0.2 degrees,
or 12.3 +/-0.2 degrees, 21.7 +/-0.2 degrees, 24.7 +/-0.2 degrees, 11.0 +/-0.2 degrees and 15.2 +/-0.2 degrees,
or 22.8 +/-0.2 degrees, 19.9 +/-0.2 degrees, 18.3 +/-0.2 degrees, 12.3 +/-0.2 degrees, 21.7 +/-0.2 degrees and 15.2 +/-0.2 degrees,
or 19.9 +/-0.2 degrees, 18.3 +/-0.2 degrees, 12.3 +/-0.2 degrees, 21.7 +/-0.2 degrees, 24.7 +/-0.2 degrees and 14.3 +/-0.2 degrees,
or 18.3 +/-0.2 degrees, 12.3 +/-0.2 degrees, 21.7 +/-0.2 degrees, 24.7 +/-0.2 degrees, 11.0 +/-0.2 degrees, 15.2 +/-0.2 degrees, 27.4 +/-0.2 degrees and 20.2 +/-0.2 degrees,
or 12.3 +/-0.2 degrees, 21.7 +/-0.2 degrees, 24.7 +/-0.2 degrees, 11.0 +/-0.2 degrees, 15.2 +/-0.2 degrees, 27.4 +/-0.2 degrees, 20.2 +/-0.2 degrees and 14.3 +/-0.2 degrees,
or 22.8 +/-0.2 degrees, 19.9 +/-0.2 degrees, 18.3 +/-0.2 degrees, 12.3 +/-0.2 degrees, 21.7 +/-0.2 degrees, 24.7 +/-0.2 degrees, 11.0 +/-0.2 degrees, 15.2 +/-0.2 degrees, 27.4 +/-0.2 degrees and 11.0 +/-0.2 degrees,
or 19.9 +/-0.2 degrees, 18.3 +/-0.2 degrees, 12.3 +/-0.2 degrees, 21.7 +/-0.2 degrees, 24.7 +/-0.2 degrees, 11.0 +/-0.2 degrees, 15.2 +/-0.2 degrees, 27.4 +/-0.2 degrees, 20.2 +/-0.2 degrees and 14.3 +/-0.2 degrees.
The succinate crystal form A of the compound (S) -8- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) pyrazin-2-yl) -2-cyclopropyl-8-azaspiro [4.5] dec-2-en-1-amine of the invention has characteristic X-ray diffraction peaks expressed by 2 theta angles and interplanar distances d using Cu-Ka radiation as shown in Table 14.
TABLE 14
Figure BDA0003005435270000321
Figure BDA0003005435270000331
The succinate crystal form A of the compound (S) -8- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) pyrazin-2-yl) -2-cyclopropyl-8-azaspiro [4.5] dec-2-en-1-amine has an X-ray powder diffraction pattern basically as shown in figure 35; the DSC pattern is basically shown in figure 36.
In a further preferred embodiment of the present invention there is provided crystalline form A of the hydrobromide salt of the compound (S) -8- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) pyrazin-2-yl) -2-cyclopropyl-8-azaspiro [4.5] dec-2-en-1-amine,
wherein the compound (S) -8- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) pyrazin-2-yl) -2-cyclopropyl-8-azaspiro [4.5] dec-2-en-1-amine hydrobromide form a has an X-ray powder diffraction pattern with diffraction peaks at 21.9 ± 0.2 °; or a diffraction peak at 20.3 ± 0.2 °; or a diffraction peak at 18.3 ± 0.2 °; or a diffraction peak at 12.0 ± 0.2 °; or a diffraction peak at 24.6 ± 0.2 °; or a diffraction peak at 16.8 ± 0.2 °; or a diffraction peak at 19.7 ± 0.2 °; or a diffraction peak at 21.4 ± 0.2 °; or a diffraction peak at 14.2 ± 0.2 °; or a diffraction peak at 32.2 ± 0.2 °; preferably any 2-5, or 3-6, or 3-8, or 5-8, or 6-8 of the above diffraction peaks, more preferably any 6, 7 or 8 thereof;
Preferably, the compound (S) -8- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) pyrazin-2-yl) -2-cyclopropyl-8-azaspiro [4.5] dec-2-en-1-amine hydrobromide form a has an X-ray powder diffraction pattern comprising at least one or more diffraction peaks, preferably 2, more preferably 3, located at 2 Θ of 21.9 ± 0.2 °, 20.3 ± 0.2 °, 18.3 ± 0.2 °; optionally, the compound further comprises at least one position with 2 theta of 12.0 +/-0.2 degrees, 24.6 +/-0.2 degrees, 16.8 +/-0.2 degrees, 19.7 +/-0.2 degrees and 21.4 +/-0.2 degrees, preferably comprises 2, 3, 4 or 5 positions,
for example, the X-ray powder diffraction pattern of the hydrobromide form a has diffraction peaks at the following positions 2 θ:
21.9 +/-0.2 degrees and 20.3 +/-0.2 degrees,
or 20.3 +/-0.2 degrees and 18.3 +/-0.2 degrees,
or 21.9 +/-0.2 degrees, 20.3 +/-0.2 degrees and 18.3 +/-0.2 degrees,
or 20.3 +/-0.2 degrees, 18.3 +/-0.2 degrees and 12.0 +/-0.2 degrees,
or 21.9 +/-0.2 degrees, 20.3 +/-0.2 degrees, 18.3 +/-0.2 degrees and 12.0 +/-0.2 degrees,
or 20.3 +/-0.2 degrees, 18.3 +/-0.2 degrees, 12.0 +/-0.2 degrees and 24.6 +/-0.2 degrees,
or 21.9 +/-0.2 degrees, 20.3 +/-0.2 degrees, 18.3 +/-0.2 degrees, 12.0 +/-0.2 degrees and 24.6 +/-0.2 degrees,
or 20.3 +/-0.2 degrees, 18.3 +/-0.2 degrees, 12.0 +/-0.2 degrees, 24.6 +/-0.2 degrees and 16.8 +/-0.2 degrees,
More preferably, the X-ray powder diffraction pattern of the hydrobromide form a of the compound (S) -8- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) pyrazin-2-yl) -2-cyclopropyl-8-azaspiro [4.5] dec-2-en-1-amine optionally further comprises one or more diffraction peaks located at 2 Θ of 14.2 ± 0.2 °, 32.2 ± 0.2 °, 17.5 ± 0.2 °, 22.4 ± 0.2 °, 25.8 ± 0.2 °, 12.4 ± 0.2 °, 13.0 ± 0.2 °; preferably at least any 2-3, or 4-5, or 6-7 thereof; further preferably, any 2, 3, 4, 5, 6, 7,
for example, the X-ray powder diffraction pattern of the hydrobromide form a has diffraction peaks at the following positions 2 θ:
14.2 +/-0.2 degrees, 32.2 +/-0.2 degrees, 17.5 +/-0.2 degrees, 22.4 +/-0.2 degrees, 25.8 +/-0.2 degrees and 12.4 +/-0.2 degrees,
or 32.2 +/-0.2 degrees, 17.5 +/-0.2 degrees, 22.4 +/-0.2 degrees, 25.8 +/-0.2 degrees, 12.4 +/-0.2 degrees and 13.0 +/-0.2 degrees,
or 14.2 +/-0.2 degrees, 32.2 +/-0.2 degrees, 22.4 +/-0.2 degrees, 25.8 +/-0.2 degrees, 12.4 +/-0.2 degrees and 13.0 +/-0.2 degrees,
further preferably, the compound (S) -8- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) pyrazin-2-yl) -2-cyclopropyl-8-azaspiro [4.5] dec-2-ene-1-amine hydrobromide form a has an X-ray powder diffraction pattern comprising diffraction peaks at one or more of the positions of 21.9 ± 0.2 °, 20.3 ± 0.2 °, 18.3 ± 0.2 °, 12.0 ± 0.2 °, 24.6 ± 0.2 °, 16.8 ± 0.2 °, 19.7 ± 0.2 °, 21.4 ± 0.2 °, 14.2 ± 0.2 °, 32.2 ± 0.2 °, 17.5 ± 0.2 °, 22.4 ± 0.2 °, 25.8 ± 0.2 °, 12.4 ± 0.2 °, 13.0 ± 0.2 ° 2 θ; preferably, the compound contains diffraction peaks at 4, 5, 6, 8 and 10,
For example, the X-ray powder diffraction pattern of the hydrobromide form a has diffraction peaks at the following positions 2 θ:
21.9 +/-0.2 degrees, 18.3 +/-0.2 degrees, 12.0 +/-0.2 degrees and 24.6 +/-0.2 degrees,
or 20.3 +/-0.2 degrees, 24.6 +/-0.2 degrees, 16.8 +/-0.2 degrees and 19.7 +/-0.2 degrees,
or 18.3 +/-0.2 degrees, 12.0 +/-0.2 degrees, 24.6 +/-0.2 degrees, 16.8 +/-0.2 degrees and 21.4 +/-0.2 degrees,
or 12.0 +/-0.2 degrees, 24.6 +/-0.2 degrees, 16.8 +/-0.2 degrees, 19.7 +/-0.2 degrees and 14.2 +/-0.2 degrees,
or 21.9 +/-0.2 degrees, 20.3 +/-0.2 degrees, 18.3 +/-0.2 degrees, 12.0 +/-0.2 degrees, 16.8 +/-0.2 degrees and 19.7 +/-0.2 degrees,
or 20.3 +/-0.2 degrees, 12.0 +/-0.2 degrees, 24.6 +/-0.2 degrees, 16.8 +/-0.2 degrees, 19.7 +/-0.2 degrees and 32.2 +/-0.2 degrees,
or 18.3 +/-0.2 degrees, 12.0 +/-0.2 degrees, 24.6 +/-0.2 degrees, 16.8 +/-0.2 degrees, 19.7 +/-0.2 degrees, 21.4 +/-0.2 degrees, 14.2 +/-0.2 degrees and 17.5 +/-0.2 degrees,
or 12.0 +/-0.2 degrees, 24.6 +/-0.2 degrees, 16.8 +/-0.2 degrees, 19.7 +/-0.2 degrees, 21.4 +/-0.2 degrees, 14.2 +/-0.2 degrees, 32.2 +/-0.2 degrees and 17.5 +/-0.2 degrees,
or 21.9 +/-0.2 degrees, 20.3 +/-0.2 degrees, 18.3 +/-0.2 degrees, 12.0 +/-0.2 degrees, 24.6 +/-0.2 degrees, 16.8 +/-0.2 degrees, 19.7 +/-0.2 degrees, 21.4 +/-0.2 degrees, 14.2 +/-0.2 degrees and 32.2 +/-0.2 degrees,
or 20.3 +/-0.2 degrees, 18.3 +/-0.2 degrees, 12.0 +/-0.2 degrees, 24.6 +/-0.2 degrees, 16.8 +/-0.2 degrees, 19.7 +/-0.2 degrees, 21.4 +/-0.2 degrees, 14.2 +/-0.2 degrees, 32.2 +/-0.2 degrees and 17.5 +/-0.2 degrees.
The characteristic diffraction peaks of X-rays of a hydrobromide salt of the compound (S) -8- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) pyrazin-2-yl) -2-cyclopropyl-8-azaspiro [4.5] dec-2-en-1-amine of the present invention, expressed as 2 theta angles and interplanar spacings d values using Cu-Ka radiation, are shown in Table 15.
Watch 15
Figure BDA0003005435270000351
Figure BDA0003005435270000361
The compound of the invention, namely (S) -8- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) pyrazin-2-yl) -2-cyclopropyl-8-azaspiro [4.5] dec-2-en-1-amine, is in a hydrobromide crystal form A, and has an X-ray powder diffraction pattern basically as shown in a figure 37; the DSC pattern is basically shown in figure 38.
In a further preferred embodiment of the present invention, there is provided the compound (S) -8- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) pyrazin-2-yl) -2-cyclopropyl-8-azaspiro [4.5] dec-2-en-1-amine hydrochloride form a,
wherein the compound (S) -8- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) pyrazin-2-yl) -2-cyclopropyl-8-azaspiro [4.5] dec-2-en-1-amine hydrochloride form a has an X-ray powder diffraction pattern with diffraction peaks at 22.5 ± 0.2 °; or a diffraction peak at 20.2 ± 0.2 °; or a diffraction peak at 12.2 ± 0.2 °; or a diffraction peak at 17.0 ± 0.2 °; or a diffraction peak at 28.7 ± 0.2 °; or a diffraction peak at 21.4 ± 0.2 °; or a diffraction peak at 18.7 ± 0.2 °; or a diffraction peak at 14.5 ± 0.2 °; or a diffraction peak at 18.0 ± 0.2 °; or a diffraction peak at 25.5 ± 0.2 °; preferably any 2-5, or 3-6, or 3-8, or 5-8, or 6-8 of the above diffraction peaks, more preferably any 6, 7 or 8 thereof;
Preferably, the compound (S) -8- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) pyrazin-2-yl) -2-cyclopropyl-8-azaspiro [4.5] dec-2-en-1-amine hydrochloride form a has an X-ray powder diffraction pattern comprising at least one or more diffraction peaks, preferably 2, more preferably 3, located at 2 Θ of 22.5 ± 0.2 °, 20.2 ± 0.2 °, 12.2 ± 0.2 °; optionally, the compound further comprises at least one position with 2 theta of 17.0 +/-0.2 degrees, 28.7 +/-0.2 degrees, 21.4 +/-0.2 degrees, 18.7 +/-0.2 degrees and 14.5 +/-0.2 degrees, preferably comprises 2, 3, 4 or 5 positions,
for example, the X-ray powder diffraction pattern of the hydrochloride form a has diffraction peaks at the following positions 2 θ:
22.5 plus or minus 0.2 degrees and 20.2 plus or minus 0.2 degrees,
or 20.2 +/-0.2 degrees and 12.2 +/-0.2 degrees,
or at 22.5 +/-0.2 degrees, 20.2 +/-0.2 degrees and 12.2 +/-0.2 degrees,
or 20.2 +/-0.2 degrees, 12.2 +/-0.2 degrees and 17.0 +/-0.2 degrees,
or 22.5 +/-0.2 degrees, 20.2 +/-0.2 degrees, 12.2 +/-0.2 degrees and 17.0 +/-0.2 degrees,
or 20.2 +/-0.2 degrees, 12.2 +/-0.2 degrees, 17.0 +/-0.2 degrees and 28.7 +/-0.2 degrees,
or 22.5 +/-0.2 degrees, 20.2 +/-0.2 degrees, 12.2 +/-0.2 degrees, 17.0 +/-0.2 degrees and 28.7 +/-0.2 degrees,
or 20.2 +/-0.2 degrees, 12.2 +/-0.2 degrees, 17.0 +/-0.2 degrees, 28.7 +/-0.2 degrees and 21.4 +/-0.2 degrees,
More preferably, the compound (S) -8- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) pyrazin-2-yl) -2-cyclopropyl-8-azaspiro [4.5] dec-2-en-1-amine hydrochloride form a optionally further comprises one or more diffraction peaks at 2 Θ of 18.0 ± 0.2 °, 25.5 ± 0.2 °, 28.3 ± 0.2 °, 23.8 ± 0.2 °, 12.6 ± 0.2 °, 24.8 ± 0.2 °, 13.3 ± 0.2 °; preferably at least any 2-3, or 4-5, or 6-7 thereof; further preferably, any 2, 3, 4, 5, 6, 7,
for example, the X-ray powder diffraction pattern of the hydrochloride form a has diffraction peaks at the following positions 2 θ:
18.0 +/-0.2 degrees, 25.5 +/-0.2 degrees, 28.3 +/-0.2 degrees, 23.8 +/-0.2 degrees, 12.6 +/-0.2 degrees and 24.8 +/-0.2 degrees,
or 25.5 +/-0.2 degrees, 28.3 +/-0.2 degrees, 23.8 +/-0.2 degrees, 12.6 +/-0.2 degrees, 24.8 +/-0.2 degrees and 13.3 +/-0.2 degrees,
or 18.0 +/-0.2 degrees, 25.5 +/-0.2 degrees, 23.8 +/-0.2 degrees, 12.6 +/-0.2 degrees, 24.8 +/-0.2 degrees and 13.3 +/-0.2 degrees,
further preferably, the compound (S) -8- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) pyrazin-2-yl) -2-cyclopropyl-8-azaspiro [4.5] dec-2-ene-1-amine hydrochloride form a has an X-ray powder diffraction pattern comprising diffraction peaks at one or more of 22.5 ± 0.2 °, 20.2 ± 0.2 °, 12.2 ± 0.2 °, 17.0 ± 0.2 °, 28.7 ± 0.2 °, 21.4 ± 0.2 °, 18.7 ± 0.2 °, 14.5 ± 0.2 °, 18.0 ± 0.2 °, 25.5 ± 0.2 °, 28.3 ± 0.2 °, 23.8 ± 0.2 °, 12.6 ± 0.2 °, 24.8 ± 0.2 °, 13.3 ± 0.2 ° 2 at 2 Θ; preferably, the compound contains diffraction peaks at 4, 5, 6, 8 and 10,
For example, the X-ray powder diffraction pattern of the hydrochloride form a has diffraction peaks at the following positions 2 θ:
22.5 +/-0.2 degrees, 12.2 +/-0.2 degrees, 17.0 +/-0.2 degrees and 28.7 +/-0.2 degrees,
or 20.2 +/-0.2 degrees, 17.0 +/-0.2 degrees, 28.7 +/-0.2 degrees and 21.4 +/-0.2 degrees,
or 12.2 +/-0.2 degrees, 28.7 +/-0.2 degrees, 21.4 +/-0.2 degrees, 18.7 +/-0.2 degrees and 14.5 +/-0.2 degrees,
or 17.0 +/-0.2 degrees, 28.7 +/-0.2 degrees, 21.4 +/-0.2 degrees, 18.7 +/-0.2 degrees and 14.5 +/-0.2 degrees,
or 22.5 +/-0.2 degrees, 12.2 +/-0.2 degrees, 17.0 +/-0.2 degrees, 28.7 +/-0.2 degrees, 21.4 +/-0.2 degrees and 18.7 +/-0.2 degrees,
or 20.2 +/-0.2 degrees, 17.0 +/-0.2 degrees, 28.7 +/-0.2 degrees, 21.4 +/-0.2 degrees, 18.7 +/-0.2 degrees and 14.5 +/-0.2 degrees,
or 12.2 +/-0.2 degrees, 17.0 +/-0.2 degrees, 28.7 +/-0.2 degrees, 21.4 +/-0.2 degrees, 18.7 +/-0.2 degrees, 14.5 +/-0.2 degrees, 18.0 +/-0.2 degrees and 25.5 +/-0.2 degrees,
or 17.0 +/-0.2 degrees, 28.7 +/-0.2 degrees, 21.4 +/-0.2 degrees, 18.7 +/-0.2 degrees, 14.5 +/-0.2 degrees, 18.0 +/-0.2 degrees, 25.5 +/-0.2 degrees and 28.3 +/-0.2 degrees,
or 22.5 +/-0.2 degrees, 20.2 +/-0.2 degrees, 12.2 +/-0.2 degrees, 17.0 +/-0.2 degrees, 28.7 +/-0.2 degrees, 21.4 +/-0.2 degrees, 18.7 +/-0.2 degrees, 14.5 +/-0.2 degrees, 18.0 +/-0.2 degrees and 25.5 +/-0.2 degrees,
or 20.2 +/-0.2 degrees, 12.2 +/-0.2 degrees, 17.0 +/-0.2 degrees, 28.7 +/-0.2 degrees, 21.4 +/-0.2 degrees, 18.7 +/-0.2 degrees, 14.5 +/-0.2 degrees, 18.0 +/-0.2 degrees, 25.5 +/-0.2 degrees and 28.3 +/-0.2 degrees.
The hydrochloride form a of the compound (S) -8- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) pyrazin-2-yl) -2-cyclopropyl-8-azaspiro [4.5] dec-2-en-1-amine of the present invention has characteristic X-ray diffraction peaks expressed in terms of 2 θ angles and interplanar spacings d using Cu — ka radiation as shown in table 16.
TABLE 16
Figure BDA0003005435270000381
The hydrochloride crystal form A of the compound (S) -8- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) pyrazin-2-yl) -2-cyclopropyl-8-azaspiro [4.5] dec-2-en-1-amine has an X-ray powder diffraction pattern basically as shown in a figure 39; the DSC pattern is basically shown in figure 40.
In a further preferred embodiment of the present invention, the compound (S) -8- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) pyrazin-2-yl) -2-cyclopropyl-8-azaspiro [4.5] dec-2-en-1-amine is provided in crystalline maleate form a.
Wherein the compound (S) -8- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) pyrazin-2-yl) -2-cyclopropyl-8-azaspiro [4.5] dec-2-en-1-amine maleate salt form a has a diffraction peak at 21.4 ± 0.2 ° in the X-ray powder diffraction pattern; or a diffraction peak at 17.4 ± 0.2 °; or a diffraction peak at 20.0 ± 0.2 °; or a diffraction peak at 25.6 ± 0.2 °; or a diffraction peak at 13.8 ± 0.2 °; or a diffraction peak at 18.3 ± 0.2 °; or a diffraction peak at 16.2 ± 0.2 °; or a diffraction peak at 20.3 ± 0.2 °; or a diffraction peak at 22.9 ± 0.2 °; or a diffraction peak at 13.3 ± 0.2 °; preferably any 2-5, or 3-6, or 3-8, or 5-8, or 6-8 of the above diffraction peaks, more preferably any 6, 7 or 8 thereof;
Preferably, the compound (S) -8- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) pyrazin-2-yl) -2-cyclopropyl-8-azaspiro [4.5] dec-2-en-1-amine maleate salt form a has an X-ray powder diffraction pattern comprising at least one or more diffraction peaks, preferably 2, more preferably 3, located at 2 Θ of 21.4 ± 0.2 °, 17.4 ± 0.2 °, 20.0 ± 0.2 °; optionally, the compound further comprises at least one position with 2 theta of 25.6 +/-0.2 degrees, 13.8 +/-0.2 degrees, 18.3 +/-0.2 degrees, 16.2 +/-0.2 degrees and 20.3 +/-0.2 degrees, preferably comprises 2, 3, 4 or 5 positions,
for example, the maleate form a has an X-ray powder diffraction pattern with diffraction peaks at the following positions 2 θ:
21.4 +/-0.2 degrees and 17.4 +/-0.2 degrees,
or 17.4 +/-0.2 degrees and 20.0 +/-0.2 degrees,
or 21.4 +/-0.2 degrees, 17.4 +/-0.2 degrees and 20.0 +/-0.2 degrees,
or 17.4 +/-0.2 degrees, 20.0 +/-0.2 degrees and 25.6 +/-0.2 degrees,
or 21.4 +/-0.2 degrees, 17.4 +/-0.2 degrees, 20.0 +/-0.2 degrees and 25.6 +/-0.2 degrees,
or 17.4 +/-0.2 degrees, 20.0 +/-0.2 degrees, 25.6 +/-0.2 degrees and 13.8 +/-0.2 degrees,
or 21.4 +/-0.2 degrees, 17.4 +/-0.2 degrees, 20.0 +/-0.2 degrees, 25.6 +/-0.2 degrees and 13.8 +/-0.2 degrees,
or 17.4 +/-0.2 degrees, 20.0 +/-0.2 degrees, 25.6 +/-0.2 degrees, 13.8 +/-0.2 degrees and 18.3 +/-0.2 degrees,
More preferably, the compound (S) -8- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) pyrazin-2-yl) -2-cyclopropyl-8-azaspiro [4.5] dec-2-en-1-amine maleate salt form a optionally further comprises one or more diffraction peaks at 22.9 ± 0.2 °, 13.3 ± 0.2 °, 22.5 ± 0.2 °, 25.9 ± 0.2 °, 10.9 ± 0.2 °, 27.6 ± 0.2 °, 23.5 ± 0.2 ° 2 Θ; preferably at least any 2-3, or 4-5, or 6-7 thereof; further preferably, any 2, 3, 4, 5, 6, 7,
for example, the maleate form a has an X-ray powder diffraction pattern with diffraction peaks at the following positions 2 θ:
22.9 +/-0.2 degrees, 13.3 +/-0.2 degrees, 22.5 +/-0.2 degrees, 25.9 +/-0.2 degrees, 10.9 +/-0.2 degrees and 27.6 +/-0.2 degrees,
or 13.3 +/-0.2 degrees, 22.5 +/-0.2 degrees, 25.9 +/-0.2 degrees, 10.9 +/-0.2 degrees, 27.6 +/-0.2 degrees and 23.5 +/-0.2 degrees,
or 22.9 +/-0.2 degrees, 13.3 +/-0.2 degrees, 25.9 +/-0.2 degrees, 10.9 +/-0.2 degrees, 27.6 +/-0.2 degrees and 23.5 +/-0.2 degrees,
further preferably, the compound (S) -8- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) pyrazin-2-yl) -2-cyclopropyl-8-azaspiro [4.5] dec-2-en-1-amine maleate salt form a has an X-ray powder diffraction pattern comprising diffraction peaks at one or more of the positions of 21.4 ± 0.2 °, 17.4 ± 0.2 °, 20.0 ± 0.2 °, 25.6 ± 0.2 °, 13.8 ± 0.2 °, 18.3 ± 0.2 °, 16.2 ± 0.2 °, 20.3 ± 0.2 °, 22.9 ± 0.2 °, 13.3 ± 0.2 °, 22.5 ± 0.2 °, 25.9 ± 0.2 °, 10.9 ± 0.2 °, 27.6 ± 0.2 °, 23.5 ± 0.2 ° 2; preferably, the compound contains diffraction peaks at 4, 5, 6, 8 and 10,
For example, the maleate form a has an X-ray powder diffraction pattern with diffraction peaks at the following positions 2 θ:
21.4 +/-0.2 degrees, 20.0 +/-0.2 degrees, 25.6 +/-0.2 degrees and 13.8 +/-0.2 degrees,
or 17.4 +/-0.2 degrees, 25.6 +/-0.2 degrees, 13.8 +/-0.2 degrees and 18.3 +/-0.2 degrees,
or 20.0 +/-0.2 degrees, 13.8 +/-0.2 degrees, 18.3 +/-0.2 degrees, 16.2 +/-0.2 degrees and 20.3 +/-0.2 degrees,
or 25.6 +/-0.2 degrees, 18.3 +/-0.2 degrees, 16.2 +/-0.2 degrees, 20.3 +/-0.2 degrees and 22.9 +/-0.2 degrees,
or 21.4 +/-0.2 degrees, 20.0 +/-0.2 degrees, 25.6 +/-0.2 degrees, 13.8 +/-0.2 degrees, 18.3 +/-0.2 degrees and 16.2 +/-0.2 degrees,
or 17.4 +/-0.2 degrees, 25.6 +/-0.2 degrees, 13.8 +/-0.2 degrees, 18.3 +/-0.2 degrees, 16.2 +/-0.2 degrees and 20.3 +/-0.2 degrees,
or 20.0 +/-0.2 degrees, 13.8 +/-0.2 degrees, 18.3 +/-0.2 degrees, 16.2 +/-0.2 degrees, 20.3 +/-0.2 degrees, 22.9 +/-0.2 degrees, 13.3 +/-0.2 degrees and 22.5 +/-0.2 degrees,
or 25.6 +/-0.2 degrees, 18.3 +/-0.2 degrees, 16.2 +/-0.2 degrees, 20.3 +/-0.2 degrees, 22.9 +/-0.2 degrees, 13.3 +/-0.2 degrees, 22.5 +/-0.2 degrees and 25.9 +/-0.2 degrees,
or 21.4 +/-0.2 degrees, 17.4 +/-0.2 degrees, 20.0 +/-0.2 degrees, 25.6 +/-0.2 degrees, 13.8 +/-0.2 degrees, 18.3 +/-0.2 degrees, 16.2 +/-0.2 degrees, 20.3 +/-0.2 degrees, 22.9 +/-0.2 degrees and 13.3 +/-0.2 degrees,
or 17.4 +/-0.2 degrees, 20.0 +/-0.2 degrees, 25.6 +/-0.2 degrees, 13.8 +/-0.2 degrees, 18.3 +/-0.2 degrees, 16.2 +/-0.2 degrees, 20.3 +/-0.2 degrees, 22.9 +/-0.2 degrees, 13.3 +/-0.2 degrees and 22.5 +/-0.2 degrees.
The compound of the invention, i.e. (S) -8- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) pyrazin-2-yl) -2-cyclopropyl-8-azaspiro [4.5] dec-2-en-1-amine, is in the form of maleate crystal form A, and X-ray characteristic diffraction peaks expressed in terms of 2 theta angles and interplanar distances d using Cu-Ka radiation are shown in Table 17.
TABLE 17
Figure BDA0003005435270000401
Figure BDA0003005435270000411
The compound of the invention, namely (S) -8- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) pyrazin-2-yl) -2-cyclopropyl-8-azaspiro [4.5] dec-2-en-1-amine, is in a maleate crystal form A, and the X-ray powder diffraction pattern of the maleate crystal form A is basically shown in figure 41; the DSC pattern is basically shown in figure 42.
In a further preferred embodiment of the present invention there is provided crystalline form a of the tartrate salt of the compound (S) -8- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) pyrazin-2-yl) -2-cyclopropyl-8-azaspiro [4.5] dec-2-en-1-amine,
wherein the X-ray powder diffraction pattern of the tartrate crystal form A has a diffraction peak at 20.1 +/-0.2 degrees; or a diffraction peak at 22.8 ± 0.2 °; or a diffraction peak at 12.6 ± 0.2 °; or a diffraction peak at 18.4 ± 0.2 °; or a diffraction peak at 18.1 ± 0.2 °; or a diffraction peak at 24.5 ± 0.2 °; or a diffraction peak at 11.2 ± 0.2 °; or a diffraction peak at 29.0 ± 0.2 °; or a diffraction peak at 20.7 ± 0.2 °; or a diffraction peak at 21.6 ± 0.2 °; preferably any 2-5, or 3-6, or 3-8, or 5-8, or 6-8 of the above diffraction peaks, more preferably any 6, 7 or 8 thereof;
Preferably, the X-ray powder diffraction pattern of tartrate form a comprises at least one or more diffraction peaks, preferably 2, more preferably 3, located at 2 Θ of 20.1 ± 0.2 °, 22.8 ± 0.2 °, 12.6 ± 0.2 °; optionally, the compound further comprises at least one position with 2 theta of 18.4 +/-0.2 degrees, 18.1 +/-0.2 degrees, 24.5 +/-0.2 degrees, 11.2 +/-0.2 degrees and 29.0 +/-0.2 degrees, preferably comprises 2, 3, 4 or 5 positions,
for example, the tartrate form a has an X-ray powder diffraction pattern with diffraction peaks at the following positions 2 θ:
at 20.1 +/-0.2 degrees and 22.8 +/-0.2 degrees,
or at 22.8 +/-0.2 degrees and 12.6 +/-0.2 degrees,
or 20.1 +/-0.2 degrees, 22.8 +/-0.2 degrees and 12.6 +/-0.2 degrees,
or at 22.8 +/-0.2 degrees, 12.6 +/-0.2 degrees and 18.4 +/-0.2 degrees,
or 20.1 +/-0.2 degrees, 22.8 +/-0.2 degrees, 12.6 +/-0.2 degrees and 18.4 +/-0.2 degrees,
or at 22.8 +/-0.2 degrees, 12.6 +/-0.2 degrees, 18.4 +/-0.2 degrees and 18.1 +/-0.2 degrees,
or 20.1 +/-0.2 degrees, 22.8 +/-0.2 degrees, 12.6 +/-0.2 degrees, 18.4 +/-0.2 degrees and 18.1 +/-0.2 degrees,
or 22.8 +/-0.2 degrees, 12.6 +/-0.2 degrees, 18.4 +/-0.2 degrees, 18.1 +/-0.2 degrees and 24.5 +/-0.2 degrees,
more preferably, the X-ray powder diffraction pattern of tartrate form a optionally further comprises one or more diffraction peaks at 20.7 ± 0.2 °, 21.6 ± 0.2 °, 33.7 ± 0.2 °, 22.1 ± 0.2 °, 13.9 ± 0.2 °, 15.3 ± 0.2 °, 27.4 ± 0.2 ° 2 Θ; preferably at least any 2-3, or 4-5, or 6-7 thereof; further preferably, any 2, 3, 4, 5, 6, 7,
For example, the tartrate form a has an X-ray powder diffraction pattern with diffraction peaks at the following positions 2 θ:
20.7 +/-0.2 degrees, 21.6 +/-0.2 degrees, 33.7 +/-0.2 degrees, 22.1 +/-0.2 degrees, 13.9 +/-0.2 degrees and 15.3 +/-0.2 degrees,
or 21.6 +/-0.2 degrees, 33.7 +/-0.2 degrees, 22.1 +/-0.2 degrees, 13.9 +/-0.2 degrees, 15.3 +/-0.2 degrees and 27.4 +/-0.2 degrees,
or 20.7 +/-0.2 degrees, 21.6 +/-0.2 degrees, 33.7 +/-0.2 degrees, 13.9 +/-0.2 degrees, 15.3 +/-0.2 degrees and 27.4 +/-0.2 degrees,
further preferably, the X-ray powder diffraction pattern of tartrate form a comprises diffraction peaks at one or more of 20.1 ± 0.2 °, 22.8 ± 0.2 °, 12.6 ± 0.2 °, 18.4 ± 0.2 °, 18.1 ± 0.2 °, 24.5 ± 0.2 °, 11.2 ± 0.2 °, 29.0 ± 0.2 °, 20.7 ± 0.2 °, 21.6 ± 0.2 °, 33.7 ± 0.2 °, 22.1 ± 0.2 °, 13.9 ± 0.2 °, 15.3 ± 0.2 °, and 27.4 ± 0.2 ° of 2 Θ; preferably, the compound contains diffraction peaks at 4, 5, 6, 8 and 10,
for example, the tartrate form a has an X-ray powder diffraction pattern with diffraction peaks at the following positions 2 θ:
20.1 +/-0.2 degrees, 12.6 +/-0.2 degrees, 18.4 +/-0.2 degrees and 18.1 +/-0.2 degrees,
or 22.8 +/-0.2 degrees, 18.4 +/-0.2 degrees, 18.1 +/-0.2 degrees and 24.5 +/-0.2 degrees,
or 12.6 +/-0.2 degrees, 24.5 +/-0.2 degrees, 11.2 +/-0.2 degrees, 29.0 +/-0.2 degrees and 20.7 +/-0.2 degrees,
Or 18.4 +/-0.2 degrees, 24.5 +/-0.2 degrees, 11.2 +/-0.2 degrees, 29.0 +/-0.2 degrees and 20.7 +/-0.2 degrees,
or 20.1 +/-0.2 degrees, 12.6 +/-0.2 degrees, 18.4 +/-0.2 degrees, 18.1 +/-0.2 degrees, 24.5 +/-0.2 degrees and 11.2 +/-0.2 degrees,
or 22.8 +/-0.2 degrees, 18.4 +/-0.2 degrees, 18.1 +/-0.2 degrees, 24.5 +/-0.2 degrees, 11.2 +/-0.2 degrees and 29.0 +/-0.2 degrees,
or 20.1 +/-0.2 degrees, 22.8 +/-0.2 degrees, 12.6 +/-0.2 degrees, 18.4 +/-0.2 degrees, 18.1 +/-0.2 degrees, 24.5 +/-0.2 degrees, 11.2 +/-0.2 degrees and 29.0 +/-0.2 degrees,
or 22.8 +/-0.2 degrees, 12.6 +/-0.2 degrees, 18.4 +/-0.2 degrees, 18.1 +/-0.2 degrees, 24.5 +/-0.2 degrees, 11.2 +/-0.2 degrees, 29.0 +/-0.2 degrees and 20.7 +/-0.2 degrees,
or 20.1 +/-0.2 degrees, 22.8 +/-0.2 degrees, 12.6 +/-0.2 degrees, 18.4 +/-0.2 degrees, 18.1 +/-0.2 degrees, 24.5 +/-0.2 degrees, 11.2 +/-0.2 degrees, 29.0 +/-0.2 degrees, 20.7 +/-0.2 degrees and 21.6 +/-0.2 degrees,
or 22.8 +/-0.2 degrees, 12.6 +/-0.2 degrees, 18.4 +/-0.2 degrees, 18.1 +/-0.2 degrees, 24.5 +/-0.2 degrees, 11.2 +/-0.2 degrees, 29.0 +/-0.2 degrees, 20.7 +/-0.2 degrees, 21.6 +/-0.2 degrees and 33.7 +/-0.2 degrees.
The tartrate crystal form a of the compound (S) -8- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) pyrazin-2-yl) -2-cyclopropyl-8-azaspiro [4.5] dec-2-en-1-amine of the present invention has X-ray characteristic diffraction peaks expressed in terms of 2 theta angles and interplanar spacings d values using Cu-ka radiation as shown in table 18.
Watch 18
Figure BDA0003005435270000431
The tartrate crystal form A of the compound (S) -8- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) pyrazin-2-yl) -2-cyclopropyl-8-azaspiro [4.5] dec-2-en-1-amine has an X-ray powder diffraction pattern basically as shown in figure 43; the DSC pattern is basically shown in figure 44.
In a further preferred embodiment of the present invention, there is provided crystalline form a of the sulfate salt of the compound (S) -8- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) pyrazin-2-yl) -2-cyclopropyl-8-azaspiro [4.5] dec-2-en-1-amine.
Wherein the compound (S) -8- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) pyrazin-2-yl) -2-cyclopropyl-8-azaspiro [4.5] dec-2-en-1-amine sulfate form a has an X-ray powder diffraction pattern with diffraction peaks at 21.6 ± 0.2 °; or a diffraction peak at 23.8 ± 0.2 °; or a diffraction peak at 17.5 ± 0.2 °; or a diffraction peak at 19.7 ± 0.2 °; or a diffraction peak at 19.0 ± 0.2 °; or a diffraction peak at 10.5 ± 0.2 °; or a diffraction peak at 26.1 ± 0.2 °; or a diffraction peak at 18.5 ± 0.2 °; or a diffraction peak at 22.5 ± 0.2 °; or a diffraction peak at 13.5 ± 0.2 °; preferably any 2-5, or 3-6, or 3-8, or 5-8, or 6-8 of the above diffraction peaks, more preferably any 6, 7 or 8 thereof;
Preferably, the compound (S) -8- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) pyrazin-2-yl) -2-cyclopropyl-8-azaspiro [4.5] dec-2-en-1-amine sulfate form a has an X-ray powder diffraction pattern comprising at least one or more diffraction peaks, preferably 2, more preferably 3, located at 2 Θ of 21.6 ± 0.2 °, 23.8 ± 0.2 °, 17.5 ± 0.2 °; optionally, the compound further comprises at least one position with 2 theta of 19.7 +/-0.2 degrees, 19.0 +/-0.2 degrees, 10.5 +/-0.2 degrees, 26.1 +/-0.2 degrees and 18.5 +/-0.2 degrees, preferably comprises 2, 3, 4 or 5 positions,
for example, the X-ray powder diffraction pattern of the sulfate form a has diffraction peaks at the following positions 2 θ:
21.6 +/-0.2 degrees and 23.8 +/-0.2 degrees,
or at 23.8 +/-0.2 degrees and 17.5 +/-0.2 degrees,
or 21.6 +/-0.2 degrees, 23.8 +/-0.2 degrees and 17.5 +/-0.2 degrees,
or at 23.8 +/-0.2 degrees, 17.5 +/-0.2 degrees and 19.7 +/-0.2 degrees,
or 21.6 +/-0.2 degrees, 23.8 +/-0.2 degrees, 17.5 +/-0.2 degrees and 19.7 +/-0.2 degrees,
or at 23.8 +/-0.2 degrees, 17.5 +/-0.2 degrees, 19.7 +/-0.2 degrees and 19.0 +/-0.2 degrees,
or 21.6 +/-0.2 degrees, 23.8 +/-0.2 degrees, 17.5 +/-0.2 degrees, 19.7 +/-0.2 degrees and 19.0 +/-0.2 degrees,
or at 23.8 + -0.2 deg., 17.5 + -0.2 deg., 19.7 + -0.2 deg., 19.0 + -0.2 deg. and 10.5 + -0.2 deg.,
More preferably, the compound (S) -8- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) pyrazin-2-yl) -2-cyclopropyl-8-azaspiro [4.5] dec-2-en-1-amine sulfate form a optionally further comprises one or more diffraction peaks at 22.5 ± 0.2 °, 13.5 ± 0.2 °, 14.0 ± 0.2 °, 20.6 ± 0.2 °, 12.3 ± 0.2 °, 20.9 ± 0.2 °, 31.7 ± 0.2 ° 2 Θ; preferably at least any 2-3, or 4-5, or 6-7 thereof; further preferably, any 2, 3, 4, 5, 6, 7,
for example, the X-ray powder diffraction pattern of the sulfate form a has diffraction peaks at the following positions 2 θ:
22.5 +/-0.2 degrees, 13.5 +/-0.2 degrees, 14.0 +/-0.2 degrees, 20.6 +/-0.2 degrees, 12.3 +/-0.2 degrees and 20.9 +/-0.2 degrees,
or 13.5 +/-0.2 degrees, 14.0 +/-0.2 degrees, 20.6 +/-0.2 degrees, 12.3 +/-0.2 degrees, 20.9 +/-0.2 degrees and 31.7 +/-0.2 degrees,
or 22.5 +/-0.2 degrees, 13.5 +/-0.2 degrees, 20.6 +/-0.2 degrees, 12.3 +/-0.2 degrees, 20.9 +/-0.2 degrees and 31.7 +/-0.2 degrees,
further preferably, the compound (S) -8- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) pyrazin-2-yl) -2-cyclopropyl-8-azaspiro [4.5] dec-2-ene-1-amine sulfate form a has an X-ray powder diffraction pattern comprising diffraction peaks at one or more of 21.6 ± 0.2 °, 23.8 ± 0.2 °, 17.5 ± 0.2 °, 19.7 ± 0.2 °, 19.0 ± 0.2 °, 10.5 ± 0.2 °, 26.1 ± 0.2 °, 18.5 ± 0.2 °, 22.5 ± 0.2 °, 13.5 ± 0.2 °, 14.0 ± 0.2 °, 20.6 ± 0.2 °, 12.3 ± 0.2 °, 20.9 ± 0.2 °, 31.7 ± 0.2 ° 2 at 2 Θ; preferably, the compound contains diffraction peaks at 4, 5, 6, 8 and 10,
For example, the X-ray powder diffraction pattern of the sulfate form a has diffraction peaks at the following positions 2 θ:
21.6 +/-0.2 degrees, 17.5 +/-0.2 degrees, 19.7 +/-0.2 degrees and 19.0 +/-0.2 degrees,
or 23.8 +/-0.2 degrees, 19.7 +/-0.2 degrees, 19.0 +/-0.2 degrees and 10.5 +/-0.2 degrees,
or 17.5 +/-0.2 degrees, 19.0 +/-0.2 degrees, 10.5 +/-0.2 degrees, 26.1 +/-0.2 degrees and 18.5 +/-0.2 degrees,
or 19.7 +/-0.2 degrees, 10.5 +/-0.2 degrees, 26.1 +/-0.2 degrees, 18.5 +/-0.2 degrees and 22.5 +/-0.2 degrees,
or 21.6 +/-0.2 degrees, 17.5 +/-0.2 degrees, 19.7 +/-0.2 degrees, 19.0 +/-0.2 degrees, 10.5 +/-0.2 degrees and 26.1 +/-0.2 degrees,
or 23.8 +/-0.2 degrees, 19.7 +/-0.2 degrees, 19.0 +/-0.2 degrees, 10.5 +/-0.2 degrees, 26.1 +/-0.2 degrees and 18.5 +/-0.2 degrees,
or 17.5 +/-0.2 degrees, 19.0 +/-0.2 degrees, 10.5 +/-0.2 degrees, 26.1 +/-0.2 degrees, 18.5 +/-0.2 degrees, 22.5 +/-0.2 degrees, 13.5 +/-0.2 degrees and 14.0 +/-0.2 degrees,
or 19.7 +/-0.2 degrees, 10.5 +/-0.2 degrees, 26.1 +/-0.2 degrees, 18.5 +/-0.2 degrees, 22.5 +/-0.2 degrees, 13.5 +/-0.2 degrees, 14.0 +/-0.2 degrees and 20.6 +/-0.2 degrees,
or 21.6 +/-0.2 degrees, 23.8 +/-0.2 degrees, 17.5 +/-0.2 degrees, 19.7 +/-0.2 degrees, 19.0 +/-0.2 degrees, 10.5 +/-0.2 degrees, 26.1 +/-0.2 degrees, 18.5 +/-0.2 degrees, 22.5 +/-0.2 degrees and 13.5 +/-0.2 degrees,
or 23.8 +/-0.2 degrees, 17.5 +/-0.2 degrees, 19.7 +/-0.2 degrees, 19.0 +/-0.2 degrees, 10.5 +/-0.2 degrees, 26.1 +/-0.2 degrees, 18.5 +/-0.2 degrees, 22.5 +/-0.2 degrees, 13.5 +/-0.2 degrees and 14.0 +/-0.2 degrees.
The sulfate crystal form a of the compound (S) -8- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) pyrazin-2-yl) -2-cyclopropyl-8-azaspiro [4.5] dec-2-en-1-amine of the present invention has X-ray characteristic diffraction peaks expressed in terms of 2 θ angles and interplanar spacings d values using Cu — ka radiation as shown in table 19.
Watch 19
Figure BDA0003005435270000451
Figure BDA0003005435270000461
The compound of the invention, namely (S) -8- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) pyrazin-2-yl) -2-cyclopropyl-8-azaspiro [4.5] dec-2-en-1-amine, has a sulfate crystal form A, and an X-ray powder diffraction pattern of the crystal form A is basically shown in a figure 45; the DSC pattern is basically shown in figure 46.
The invention further provides a method for preparing the crystal form, which comprises the following steps:
1) weighing a proper amount of free alkali, and dissolving the free alkali by using a benign solvent;
2) dissolving the acid M in an organic solvent; the amount of acid M is preferably 1.2 equivalents;
3) uniformly mixing the two solutions obtained in the steps 1) and 2), volatilizing, adding a poor solvent after the solvent is volatilized, and stirring for crystallization;
4) quickly centrifuging or standing and drying to obtain a target product;
wherein:
the benign solvent of the step 1) is selected from methanol, ethanol, ethyl acetate, dichloromethane, acetone, acetonitrile, 2-butanone, 3-pentanone, tetrahydrofuran, N-dimethylformamide, N-dimethylacetamide or N-methylpyrrolidone; preferably methanol or ethanol;
The organic solvent in the step 2) is selected from methanol, ethanol, ethyl acetate, dichloromethane, acetone, N-hexane, petroleum ether, benzene, toluene, chloroform, acetonitrile, carbon tetrachloride, dichloroethane, tetrahydrofuran, 2-butanone, 3-pentanone, heptane, methyl tert-butyl ether, isopropyl ether, 1, 4-dioxane, tert-butyl alcohol or N, N-dimethylformamide; preferably methanol, ethanol or acetonitrile;
the benign solvent in the step 1) and the organic solution in the step 2) need to be mutually soluble when being used;
the poor solvent in the step 3) is selected from ethyl acetate, acetone, dichloromethane, acetonitrile, tetrahydrofuran, 2-butanone, 3-pentanone and 1, 4-dioxane; ethyl acetate, acetone or acetonitrile are preferred.
The invention further provides a method for preparing the crystal form, which comprises the following steps:
1) weighing a proper amount of compound salt, and suspending with a poor solvent to obtain a suspension; the suspension density is preferably 50-100 mg/mL;
2) shaking the suspension obtained in the step 1) for 1-10 days at 25-40 ℃;
3) quickly centrifuging the suspension obtained in the step 2), removing supernatant, and drying the residual solid in a vacuum drying oven at 40 ℃ until the weight is constant to obtain a target product;
wherein:
The poor solvent of the step 1) is selected from acetone, ethyl acetate, acetonitrile, ethanol, 88% acetone, tetrahydrofuran, 1, 4-dioxane, benzene, toluene, isopropanol, N-butanol, isobutanol, N-dimethylformamide, N-dimethylacetamide, N-propanol, tert-butanol or 2-butanone.
The invention further provides a method for preparing the crystal form, which comprises the following steps:
1) weighing a proper amount of compound salt, and dissolving the compound salt by using a benign solvent;
2) cooling the solution obtained in the step 1), stirring and precipitating solid, wherein the temperature is preferably-20-5 ℃;
3) quickly centrifuging the suspension obtained in the step 2), removing supernatant, and drying the residual solid in a vacuum drying oven at 40 ℃ to constant weight to obtain a target product;
wherein:
the benign solvent is selected from methanol, ethanol, 88% acetone, tetrahydrofuran, N-dimethylformamide, N-dimethylacetamide or N-methylpyrrolidone; methanol or 88% acetone is preferred.
The invention further provides a method for preparing the crystal form, which comprises the following steps:
1) weighing a proper amount of free alkali, and dissolving the free alkali by using a benign solvent;
2) weighing a proper amount of counter ion acid, and dissolving the counter ion acid by using an organic solvent; the amount of the counter-ionic acid is preferably 1.2 equivalents;
3) Mixing the two solutions, stirring to separate out or dripping the poor solvent 1 and stirring to separate out;
4) drying to obtain a target product;
wherein:
the benign solvent is selected from methanol, dichloromethane, 1, 4-dioxane, acetone, tetrahydrofuran, N-dimethylformamide, N-dimethylacetamide or N-methylpyrrolidone; methanol and tetrahydrofuran are preferred;
the organic solvent is selected from methanol, ethanol, ethyl acetate, dichloromethane, acetone, N-hexane, petroleum ether, benzene, toluene, chloroform, acetonitrile, carbon tetrachloride, dichloroethane, tetrahydrofuran, 2-butanone, 3-pentanone, heptane, methyl tert-butyl ether, isopropyl ether, 1, 4-dioxane, tert-butanol or N, N-dimethylformamide; preferably methanol, ethanol or acetonitrile; the benign solvent and the organic solution need to be mutually soluble when in use;
the poor solvent 1 is selected from heptane, water, methyl tert-butyl ether, toluene, isopropyl ether, ethyl acetate, acetone or acetonitrile; preferably water, methyl tert-butyl ether, isopropyl ether;
the counter-ionic acid is selected from the group consisting of hydrochloric acid, sulfuric acid, nitric acid, hydrobromic acid, hydrofluoric acid, hydroiodic acid, phosphoric acid, 2, 5-dihydroxybenzoic acid, 1-hydroxy-2-naphthoic acid, acetic acid, dichloroacetic acid, trichloroacetic acid, acetoxy-hydroxamic acid, adipic acid, benzenesulfonic acid, 4-chlorobenzenesulfonic acid, benzoic acid, 4-acetamidobenzoic acid, 4-aminobenzoic acid, capric acid, caproic acid, caprylic acid, cinnamic acid, citric acid, cyclamic acid, camphorsulfonic acid, aspartic acid, camphoric acid, gluconic acid, glucuronic acid, glutamic acid, erythorbic acid, lactic acid, malic acid, mandelic acid, pyroglutamic acid, tartaric acid, dodecylsulfuric acid, dibenzoyltartaric acid, ethane-1, 2-disulfonic acid, ethanesulfonic acid, formic acid, fumaric acid, galactonic acid, gentisic acid, glutaric acid, fumaric acid, and mixtures thereof, 2-ketoglutaric acid, glycolic acid, hippuric acid, isethionic acid, lactobionic acid, ascorbic acid, aspartic acid, lauric acid, camphoric acid, maleic acid, malonic acid, methanesulfonic acid, 1, 5-naphthalenedisulfonic acid, naphthalene-2-sulfonic acid, nicotinic acid, oleic acid, orotic acid, oxalic acid, palmitic acid, pamoic acid, propionic acid, salicylic acid, 4-aminosalicylic acid, sebacic acid, stearic acid, succinic acid, thiocyanic acid, undecylenic acid, trifluoroacetic acid, benzenesulfonic acid, p-toluenesulfonic acid, or L-malic acid; preferably fumarate, p-toluenesulfonate or succinate; most preferably p-toluenesulfonate;
Or, the method specifically comprises the following steps:
1) weighing a proper amount of free alkali, and suspending with a poor solvent 2;
2) weighing a proper amount of counter ion acid, and dissolving the counter ion acid by using an organic solvent; the amount of the counter-ionic acid is preferably 1.2 equivalents;
3) mixing the two solutions, stirring for dissolving, and continuously stirring for separating out or dropwise adding a poor solvent 3 and stirring for separating out;
4) drying to obtain a target product;
the poor solvent 2 is selected from ethyl acetate, acetone, dichloromethane, acetonitrile, tetrahydrofuran, 2-methyl-tetrahydrofuran, 2-butanone, 3-pentanone, 1, 4-dioxane and ethanol; preferably ethyl acetate, acetone, acetonitrile, 2-methyl-tetrahydrofuran, ethanol;
the organic solvent is selected from methanol, ethanol, ethyl acetate, dichloromethane, acetone, N-hexane, petroleum ether, benzene, toluene, chloroform, acetonitrile, carbon tetrachloride, dichloroethane, tetrahydrofuran, 2-butanone, 3-pentanone, heptane, methyl tert-butyl ether, isopropyl ether, 1, 4-dioxane, tert-butanol or N, N-dimethylformamide; preferably methanol, ethanol or acetonitrile; the benign solvent and the organic solution need to be mutually soluble when in use;
the poor solvent 3 is selected from heptane, water, methyl tert-butyl ether, toluene, isopropyl ether, ethyl acetate, acetone or acetonitrile; preferably water, methyl tert-butyl ether, isopropyl ether;
The counter-ionic acid is selected from the group consisting of hydrochloric acid, sulfuric acid, nitric acid, hydrobromic acid, hydrofluoric acid, hydroiodic acid, phosphoric acid, 2, 5-dihydroxybenzoic acid, 1-hydroxy-2-naphthoic acid, acetic acid, dichloroacetic acid, trichloroacetic acid, acetoxy-hydroxamic acid, adipic acid, benzenesulfonic acid, 4-chlorobenzenesulfonic acid, benzoic acid, 4-acetamidobenzoic acid, 4-aminobenzoic acid, capric acid, caproic acid, caprylic acid, cinnamic acid, citric acid, cyclamic acid, camphorsulfonic acid, aspartic acid, camphoric acid, gluconic acid, glucuronic acid, glutamic acid, erythorbic acid, lactic acid, malic acid, mandelic acid, pyroglutamic acid, tartaric acid, dodecylsulfuric acid, dibenzoyltartaric acid, ethane-1, 2-disulfonic acid, ethanesulfonic acid, formic acid, fumaric acid, galactonic acid, gentisic acid, glutaric acid, fumaric acid, and mixtures thereof, 2-ketoglutaric acid, glycolic acid, hippuric acid, isethionic acid, lactobionic acid, ascorbic acid, aspartic acid, lauric acid, camphoric acid, maleic acid, malonic acid, methanesulfonic acid, 1, 5-naphthalenedisulfonic acid, naphthalene-2-sulfonic acid, nicotinic acid, oleic acid, orotic acid, oxalic acid, palmitic acid, pamoic acid, propionic acid, salicylic acid, 4-aminosalicylic acid, sebacic acid, stearic acid, succinic acid, thiocyanic acid, undecylenic acid, trifluoroacetic acid, benzenesulfonic acid, p-toluenesulfonic acid, or L-malic acid; preferably fumarate, p-toluenesulfonate or succinate; most preferred is p-toluenesulfonate.
The invention also aims to provide a pharmaceutical composition which contains a therapeutically effective amount of the acid salt of the general formula (I) and one or more pharmaceutically acceptable carriers, diluents or excipients.
The invention also aims to provide an acid salt shown in the general formula (I) and application of a pharmaceutical composition thereof in preparation of SHP-2 inhibitor drugs.
The invention also aims to provide a pharmaceutical composition which contains a therapeutically effective amount of the crystal form of the salt shown in the general formula (I) and one or more pharmaceutically acceptable carriers, diluents or excipients.
The invention also aims to provide a crystal form of the salt shown in the general formula (I) and application of a pharmaceutical composition of the salt in preparation of SHP-2 inhibitor drugs.
The application is the application in preparing medicines or diseases for treating Noonan syndrome, leopard skin syndrome, leukemia, neuroblastoma, melanoma, esophageal cancer, head and neck tumors, breast cancer, lung cancer, colon cancer and the like; non-small cell lung cancer, esophageal cancer and head and neck tumors are preferred.
Drawings
Figure 1 is a graphical representation of the XRPD of (S) -1'- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) -3-fluoropyrazin-2-yl) -1, 3-dihydrospiro [ indene-2, 4' -piperidine ] -1-amine hydrochloride form a.
Figure 2 is a DSC representation of (S) -1'- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) -3-fluoropyrazin-2-yl) -1, 3-dihydrospiro [ indene-2, 4' -piperidine ] -1-amine hydrochloride form a.
Figure 3 is a TGA representation of (S) -1'- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) -3-fluoropyrazin-2-yl) -1, 3-dihydrospiro [ indene-2, 4' -piperidine ] -1-amine hydrochloride form a.
Figure 4 is a DSC representation of (S) -1'- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) -3-fluoropyrazin-2-yl) -1, 3-dihydrospiro [ indene-2, 4' -piperidine ] -1-amine hydrochloride form B.
Figure 5 is a TGA representation of (S) -1'- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) -3-fluoropyrazin-2-yl) -1, 3-dihydrospiro [ indene-2, 4' -piperidine ] -1-amine hydrochloride form B.
Figure 6 is an XRPD representation of (S) -1'- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) -3-fluoropyrazin-2-yl) -1, 3-dihydrospiro [ indene-2, 4' -piperidine ] -1-amine ethanesulfonate crystalline form a.
Figure 7 is a DSC representation of (S) -1'- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) -3-fluoropyrazin-2-yl) -1, 3-dihydrospiro [ indene-2, 4' -piperidine ] -1-aminoethanesulfonate crystalline form a.
Figure 8 is a TGA representation of (S) -1'- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) -3-fluoropyrazin-2-yl) -1, 3-dihydrospiro [ indene-2, 4' -piperidine ] -1-aminoethanesulfonate crystalline form a.
Figure 9 is a DSC representation of (S) -1'- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) -3-fluoropyrazin-2-yl) -1, 3-dihydrospiro [ indene-2, 4' -piperidine ] -1-amine benzenesulfonate form a.
Figure 10 is a TGA representation of (S) -1'- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) -3-fluoropyrazin-2-yl) -1, 3-dihydrospiro [ indene-2, 4' -piperidine ] -1-amine benzenesulfonate form a.
Figure 11 is a DSC representation of (S) -1'- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) -3-fluoropyrazin-2-yl) -1, 3-dihydrospiro [ indene-2, 4' -piperidine ] -1-amine fumarate crystalline form a.
Figure 12 is a TGA representation of (S) -1'- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) -3-fluoropyrazin-2-yl) -1, 3-dihydrospiro [ indene-2, 4' -piperidine ] -1-amine fumarate crystalline form a.
Figure 13 is a DSC representation of (S) -1'- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) -3-fluoropyrazin-2-yl) -1, 3-dihydrospiro [ indene-2, 4' -piperidine ] -1-amine isethionate form a.
Figure 14 is a TGA representation of (S) -1'- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) -3-fluoropyrazin-2-yl) -1, 3-dihydrospiro [ indene-2, 4' -piperidine ] -1-amine isethionate crystalline form a.
Figure 15 is a DSC representation of (S) -1'- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) -3-fluoropyrazin-2-yl) -1, 3-dihydrospiro [ indene-2, 4' -piperidine ] -1-amine oxalate form a.
Figure 16 is a TGA representation of (S) -1'- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) -3-fluoropyrazin-2-yl) -1, 3-dihydrospiro [ indene-2, 4' -piperidine ] -1-amine oxalate form a.
FIG. 17 is a spectrum of (S) -1'- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) -3-fluoropyrazin-2-yl) -1, 3-dihydrospiro [ indene-2, 4' -piperidine ] -1-aminoethanesulfonate nuclear magnetic calibration content.
FIG. 18 is an ELSD detection graphical representation of (S) -1'- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) -3-fluoropyrazin-2-yl) -1, 3-dihydrospiro [ indene-2, 4' -piperidine ] -1-amine hydrochloride salt formation content.
Figure 19 is an XRPD representation of (S) -8- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) pyrazin-2-yl) -2-cyclopropyl-8-azaspiro [4.5] dec-2-en-1-amine free base crystalline form a.
Figure 20 is a DSC-TGA representation of (S) -8- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) pyrazin-2-yl) -2-cyclopropyl-8-azaspiro [4.5] dec-2-en-1-amine as free base crystalline form a.
Figure 21 is an XRPD representation of (S) -8- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) pyrazin-2-yl) -2-cyclopropyl-8-azaspiro [4.5] dec-2-en-1-amine free base crystalline form B.
Figure 22 is a DSC representation of (S) -8- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) pyrazin-2-yl) -2-cyclopropyl-8-azaspiro [4.5] dec-2-en-1-amine free base crystalline form B.
Figure 23 is an XRPD representation of (S) -8- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) pyrazin-2-yl) -2-cyclopropyl-8-azaspiro [4.5] dec-2-en-1-amine free base crystalline form C.
Figure 24 is a DSC representation of (S) -8- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) pyrazin-2-yl) -2-cyclopropyl-8-azaspiro [4.5] dec-2-en-1-amine free base crystalline form C.
Figure 25 is an XRPD representation of (S) -8- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) pyrazin-2-yl) -2-cyclopropyl-8-azaspiro [4.5] dec-2-en-1-amine mesylate salt form a.
Figure 26 is a DSC representation of (S) -8- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) pyrazin-2-yl) -2-cyclopropyl-8-azaspiro [4.5] dec-2-en-1-amine mesylate salt form a.
Figure 27 is an XRPD pattern of (S) -8- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) pyrazin-2-yl) -2-cyclopropyl-8-azaspiro [4.5] dec-2-en-1-amine isethionate crystalline form a.
Figure 28 is a DSC representation of (S) -8- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) pyrazin-2-yl) -2-cyclopropyl-8-azaspiro [4.5] dec-2-en-1-amine isethionate crystalline form a.
Figure 29 is an XRPD pattern of (S) -8- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) pyrazin-2-yl) -2-cyclopropyl-8-azaspiro [4.5] dec-2-en-1-amine fumarate crystalline form a.
Figure 30 is a DSC representation of (S) -8- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) pyrazin-2-yl) -2-cyclopropyl-8-azaspiro [4.5] dec-2-en-1-amine fumarate crystalline form a.
Figure 31 is an XRPD pattern of (S) -8- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) pyrazin-2-yl) -2-cyclopropyl-8-azaspiro [4.5] dec-2-en-1-amine p-toluenesulfonate crystalline form a.
Figure 32 is a DSC representation of (S) -8- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) pyrazin-2-yl) -2-cyclopropyl-8-azaspiro [4.5] dec-2-en-1-amine p-toluenesulfonate crystalline form a.
Figure 33 is an XRPD representation of (S) -8- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) pyrazin-2-yl) -2-cyclopropyl-8-azaspiro [4.5] dec-2-en-1-amineethanesulfonate form a.
Figure 34 is a DSC representation of (S) -8- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) pyrazin-2-yl) -2-cyclopropyl-8-azaspiro [4.5] dec-2-en-1-amineethanesulfonate form a.
Figure 35 is an XRPD pattern of (S) -8- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) pyrazin-2-yl) -2-cyclopropyl-8-azaspiro [4.5] dec-2-en-1-amine succinate crystalline form a.
Figure 36 is a DSC representation of (S) -8- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) pyrazin-2-yl) -2-cyclopropyl-8-azaspiro [4.5] dec-2-en-1-amine succinate crystalline form a.
Figure 37 is an XRPD pattern of (S) -8- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) pyrazin-2-yl) -2-cyclopropyl-8-azaspiro [4.5] dec-2-en-1-amine hydrobromide form a.
Figure 38 is a DSC representation of (S) -8- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) pyrazin-2-yl) -2-cyclopropyl-8-azaspiro [4.5] dec-2-en-1-amine hydrobromide form a.
Figure 39 is an XRPD pattern of (S) -8- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) pyrazin-2-yl) -2-cyclopropyl-8-azaspiro [4.5] dec-2-en-1-amine hydrochloride form a.
Figure 40 is a DSC representation of (S) -8- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) pyrazin-2-yl) -2-cyclopropyl-8-azaspiro [4.5] dec-2-en-1-amine hydrochloride form a.
Figure 41 is an XRPD representation of (S) -8- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) pyrazin-2-yl) -2-cyclopropyl-8-azaspiro [4.5] dec-2-en-1-amine maleate form a.
Figure 42 is a DSC representation of (S) -8- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) pyrazin-2-yl) -2-cyclopropyl-8-azaspiro [4.5] dec-2-en-1-amine maleate form a.
Figure 43 is an XRPD pattern of (S) -8- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) pyrazin-2-yl) -2-cyclopropyl-8-azaspiro [4.5] dec-2-en-1-amine tartrate form a.
Figure 44 is a DSC representation of (S) -8- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) pyrazin-2-yl) -2-cyclopropyl-8-azaspiro [4.5] dec-2-en-1-amine tartrate form a.
Figure 45 is an XRPD representation of (S) -8- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) pyrazin-2-yl) -2-cyclopropyl-8-azaspiro [4.5] dec-2-en-1-amine sulfate form a.
Figure 46 is a DSC representation of (S) -8- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) pyrazin-2-yl) -2-cyclopropyl-8-azaspiro [4.5] dec-2-en-1-amine sulfate crystalline form a.
Detailed Description
Unless stated to the contrary, terms used in the specification and claims have the following meanings.
The term "alkyl" refers to a saturated aliphatic hydrocarbon group which is a straight or branched chain group containing 1 to 20 carbon atoms, preferably an alkyl group containing 1 to 8 carbon atoms, more preferably an alkyl group of 1 to 6 carbon atoms, and most preferably an alkyl group of 1 to 3 carbon atoms. Non-limiting examples include methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, sec-butyl, n-pentyl, 1-dimethylpropyl, 1, 2-dimethylpropyl, 2-dimethylpropyl, 1-ethylpropyl, 2-methylbutyl, 3-methylbutyl, n-hexyl, 1-ethyl-2-methylpropyl, 1, 2-trimethylpropyl, 1-dimethylbutyl, 1, 2-dimethylbutyl, 2-dimethylbutyl, 1, 3-dimethylbutyl, 2-ethylbutyl, 2-methylpentyl, 3-methylpentyl, 4-methylpentyl, 2, 3-dimethylbutyl, n-heptyl, 2-methylhexyl, 3-methylhexyl, 4-methylhexyl, 5-methylhexyl, 2, 3-dimethylpentyl, 2, 4-dimethylpentyl, 2-dimethylpentyl, 3-dimethylpentyl, 2-ethylpentyl, 3-ethylpentyl, n-octyl, 2, 3-dimethylhexyl, 2, 4-dimethylhexyl, 2, 5-dimethylhexyl, 2-dimethylhexyl, 3-dimethylhexyl, 4-dimethylhexyl, 2-ethylhexyl, 3-ethylhexyl, 4-ethylhexyl, 2-methyl-2-ethylpentyl, 2-methyl-3-ethylpentyl, n-nonyl, 2-methyl-2-ethylhexyl, 2-methyl-3-ethylhexyl, 2-dimethylpentyl, 2-dimethylhexyl, 3-dimethylpentyl, 2-ethylhexyl, 3-dimethylhexyl, 2-ethylhexyl, 2-dimethylhexyl, 2-ethylhexyl, 2-dimethylhexyl, 2-dimethylhexyl, 2-dimethylhexyl, 2-ethylhexyl, 2-ethyl, 2-2, 2-2, 2-2, or, 2, 2-diethylpentyl, n-decyl, 3-diethylhexyl, 2-diethylhexyl, and various branched isomers thereof. Alkyl groups may be substituted or unsubstituted, and when substituted, the substituent may be substituted at any available point of attachment, preferably one or more groups independently selected from alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkylamino, halo, mercapto, hydroxy, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkoxy, heterocycloalkoxy, cycloalkylthio, heterocycloalkylthio, oxo, carboxy or carboxylate, preferably methyl, ethyl, isopropyl, tert-butyl, haloalkyl, deuterated alkyl, alkoxy-substituted alkyl, hydroxy-substituted alkyl and cyano-substituted alkyl.
The term "cycloalkyl" refers to a saturated or partially unsaturated monocyclic or polycyclic cyclic hydrocarbon substituent, the cycloalkyl ring containing from 3 to 20 carbon atoms, preferably from 3 to 8 carbon atoms, more preferably from 3 to 6 carbon atoms. Non-limiting examples of monocyclic cycloalkyl groups include cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl, cyclohexenyl, cyclohexadienyl, cycloheptyl, cycloheptatrienyl, cyclooctyl, and the like; polycyclic cycloalkyl groups include spiro, fused and bridged cycloalkyl groups, preferably cyclopropyl, cyclobutyl, cyclohexyl, cyclopentyl and cycloheptyl. The cycloalkyl ring may be fused to an aryl, heteroaryl or heterocycloalkyl ring, where the ring to which the parent structure is attached is cycloalkyl, non-limiting examples of which include indanyl, tetrahydronaphthyl, benzocycloheptanyl, and the like. Cycloalkyl groups may be optionally substituted or unsubstituted, and when substituted, the substituents are preferably one or more groups independently selected from alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkylamino, halogen, mercapto, hydroxy, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkoxy, heterocycloalkoxy, cycloalkylthio, heterocycloalkylthio, oxo, carboxy or carboxylate.
The term "heterocyclyl" refers to a saturated or partially unsaturated mono-or polycyclic cyclic hydrocarbon substituent containing from 3 to 20 ring atoms wherein one or more of the ring atoms is selected from nitrogen, oxygen, or S (O)m(wherein m is an integer from 0 to 2) but excludes the ring moiety of-O-O-, -O-S-, or-S-S-, the remaining ring atoms being carbon. Preferably 3 to 12 ring atoms, of which 1 to 4 are heteroatoms; more preferably from 3 to 8 ring atoms; most preferably from 3 to 8 ring atoms. Of monocyclic heterocyclic radicalsNon-limiting examples include oxetanyl, pyrrolidinyl, pyrrolidinonyl, imidazolidinyl, tetrahydrofuranyl, tetrahydrothienyl, dihydroimidazolyl, dihydrofuryl, dihydropyrazolyl, dihydropyrrolyl, piperidinyl, piperazinyl, morpholinyl, thiomorpholinyl, homopiperazinyl, pyranyl, and the like, with oxetanyl, pyrrolidinonyl, tetrahydrofuranyl, pyrazolidinyl, morpholinyl, piperazinyl, and pyranyl being preferred. Polycyclic heterocyclic groups include spiro, fused and bridged heterocyclic groups; wherein the heterocyclic groups of the spiro, fused and bridged rings are optionally linked to other groups by single bonds, or further linked to other cycloalkyl, heterocyclic, aryl and heteroaryl groups by any two or more atoms in the ring. The heterocyclyl group may be optionally substituted or unsubstituted, and when substituted, the substituents are preferably one or more groups independently selected from alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkylamino, halogen, mercapto, hydroxy, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkoxy, heterocycloalkoxy, cycloalkylthio, heterocycloalkylthio, oxo, carboxy or carboxylate.
The term "aryl" refers to a 6 to 14 membered all carbon monocyclic or fused polycyclic (i.e., rings which share adjacent pairs of carbon atoms) group having a conjugated pi-electron system, preferably 6 to 10 membered, such as phenyl and naphthyl. More preferably phenyl. The aryl ring may be fused to a heteroaryl, heterocyclyl or cycloalkyl ring, wherein the ring attached to the parent structure is an aryl ring. The aryl group may be substituted or unsubstituted, and when substituted, the substituent is preferably one or more groups independently selected from alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkylamino, halogen, mercapto, hydroxy, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkoxy, heterocycloalkoxy, cycloalkylthio, heterocycloalkylthio, carboxy or carboxylate.
The term "heteroaryl" refers to a heteroaromatic system comprising 1 to 4 heteroatoms, 5 to 14 ring atoms, wherein the heteroatoms are selected from oxygen, sulfur and nitrogen. Heteroaryl is preferably 5 to 10 membered, more preferably 5 or 6 membered, such as imidazolyl, furyl, thienyl, thiazolyl, pyrazolyl, oxazolyl, pyrrolyl, triazolyl, tetrazolyl, pyridyl, pyrimidinyl, thiadiazole, oxadiazole, pyrazinyl and the like, preferably oxazolyl, oxadiazole, tetrazole, triazolyl, thienyl, imidazolyl, pyridyl, pyrazolyl, pyrimidinyl or thiazolyl; and further selected from oxazolyl, oxadiazole, tetrazole, triazolyl, thienyl, pyridyl, thiazolyl and pyrimidyl. The heteroaryl ring may be fused to an aryl, heterocyclyl, or cycloalkyl ring, wherein the ring joined to the parent structure is a heteroaryl ring. Heteroaryl groups may be optionally substituted or unsubstituted, and when substituted, the substituents are preferably one or more groups independently selected from alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkylamino, halogen, mercapto, hydroxyl, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkoxy, heterocycloalkoxy, cycloalkylthio, heterocycloalkylthio, carboxyl or carboxylate groups.
The term "alkoxy" refers to-O- (alkyl) and-O- (unsubstituted cycloalkyl), wherein alkyl is as defined above. Alkoxy groups having 1 to 8 carbon atoms are preferred, alkoxy groups having 1 to 6 carbon atoms are more preferred, and alkoxy groups having 1 to 3 carbon atoms are most preferred. Non-limiting examples of alkoxy groups include: methoxy, ethoxy, propoxy, butoxy, cyclopropoxy, cyclobutoxy, cyclopentyloxy, cyclohexyloxy. Alkoxy groups may be optionally substituted or unsubstituted, and when substituted, the substituents are preferably one or more groups independently selected from alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkylamino, halogen, mercapto, hydroxy, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkoxy, heterocycloalkoxy, cycloalkylthio, heterocycloalkylthio, carboxy or carboxylate groups.
"haloalkyl" refers to an alkyl group substituted with one or more halogens, wherein alkyl is as defined above.
"haloalkoxy" refers to an alkoxy group substituted with one or more halogens, wherein the alkoxy group is as defined above.
"hydroxyalkyl" refers to an alkyl group substituted with a hydroxy group, wherein alkyl is as defined above.
"alkenyl" refers to alkenyl, also known as alkenylene, wherein the alkenyl may be further substituted with other related groups, such as: alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkylamino, halogen, mercapto, hydroxyl, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkoxy, heterocycloalkoxy, cycloalkylthio, heterocycloalkylthio, carboxyl or carboxylate.
"hydroxy" refers to an-OH group.
"halogen" means fluorine, chlorine, bromine or iodine.
"amino" means-NH2。
"cyano" means-CN.
"nitro" means-NO2。
"THF" refers to tetrahydrofuran.
"EtOAc" or "EA" refers to ethyl acetate.
"DMSO" refers to dimethyl sulfoxide.
"LDA" refers to lithium diisopropylamide.
"DMAP" refers to 4-dimethylaminopyridine.
"EtMgBr" refers to ethyl magnesium bromide.
"HOSu" refers to N-hydroxysuccinimide.
"EDCl" refers to 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride.
"IPA" refers to isopropanol.
"MeOH" refers to methanol.
"EtOH" refers to ethanol.
"DMF" refers to N, N-dimethylformamide.
"DIPEA" refers to N, N-diisopropylethylamine.
"HEPES" means 4-hydroxyethylpiperazine ethanesulfonic acid.
Different terms such as "X is selected from A, B or C", "X is selected from A, B and C", "X is A, B or C", "X is A, B and C" and the like all express the same meaning, that is, X can be any one or more of A, B, C.
"optional" or "optionally" means that the subsequently described event or circumstance may, but need not, occur, and that the description includes instances where the event or circumstance occurs or does not.
"substituted" means that one or more, preferably up to 5, more preferably 1 to 3, hydrogen atoms in the group are independently substituted with a corresponding number of substituents. It goes without saying that the substituents are only in their possible chemical positions, and that the person skilled in the art is able to determine (experimentally or theoretically) possible or impossible substitutions without undue effort. For example, amino or hydroxyl groups having free hydrogen may be unstable in combination with carbon atoms having unsaturated (e.g., olefinic) bonds.
"stereoisomerism" encompasses geometric isomerism (cis-trans isomerism), optical isomerism, conformational isomerism, and the like.
All hydrogen atoms described in the present invention can be replaced by deuterium, which is an isotope thereof, and any hydrogen atom in the compound of the embodiment related to the present invention can also be replaced by a deuterium atom.
"pharmaceutical composition" means a mixture containing one or more compounds described herein or a physiologically/pharmaceutically acceptable salt or prodrug thereof in admixture with other chemical components, as well as other components such as physiologically/pharmaceutically acceptable carriers and excipients. The purpose of the pharmaceutical composition is to facilitate administration to an organism, facilitate absorption of the active ingredient and exert biological activity.
"pharmaceutically acceptable salts" refers to salts of the compounds of the present invention which are safe and effective for use in the body of a mammal and which possess the requisite biological activity.
X-ray powder diffraction pattern (XRPD), which refers to the experimentally observed diffraction pattern or parameters derived therefrom, is characterized by peak position (abscissa) and peak intensity (ordinate). Those skilled in the art will appreciate that the experimental error therein will depend on the conditions of the instrument, the sample preparation and the purity of the sample. In particular, it is well known to those skilled in the art that X-ray diffraction patterns will generally vary with the conditions of the instrument, and those skilled in the art will appreciate that a suitable error tolerance for XRPD may be: 2 theta +/-0.5 degrees; 2 theta +/-0.4 degrees; 2 theta +/-0.3 degrees; 2 theta +/-0.2 deg. It is particularly noted that the relative intensities of the X-ray diffraction patterns may also vary with the experimental conditions, so that the order of the peak intensities cannot be considered as the sole or determining factor. In addition, due to the influence of experimental factors such as sample height, an overall shift in peak angle is caused, and a certain shift is usually allowed. Thus, it will be understood by those skilled in the art that any crystalline form having the same or similar characteristic peaks as the inventive profile is within the scope of the invention.
"TGA" refers to a thermogravimetric analysis (TGA) experiment.
"DSC" refers to a Differential Scanning Calorimetry (DSC) experiment.
"HPLC" refers to High Performance Liquid Chromatography (HPLC) experiments.
"PK" refers to Pharmacokinetic (PK) experiments.
The present invention is further described below with reference to examples, which are not intended to limit the scope of the present invention.
Preparation of compounds
The structure of the compounds of the invention is determined by Nuclear Magnetic Resonance (NMR) or/and liquid mass chromatography (LC-MS). NMR chemical shifts (δ) are given in parts per million (ppm). NMR was measured using a Bruker AVANCE-400 NMR spectrometer using deuterated dimethyl sulfoxide (DMSO-d)6) Deuterated methanol (CD)3OD) and deuterated chloroform (CDCl)3) Internal standard is Tetramethylsilane (TMS).
LC-MS was measured using an Agilent 1200Infinity Series Mass spectrometer. HPLC was measured using an Agilent 1200DAD high pressure liquid chromatograph (Sunfire C18150X 4.6mm column) and a Waters 2695-2996 high pressure liquid chromatograph (Gimini C)18150X 4.6mm column).
The thin layer chromatography silica gel plate adopts a tobacco yellow sea HSGF254 or Qingdao GF254 silica gel plate, the specification adopted by TLC is 0.15 mm-0.20 mm, and the specification adopted by the thin layer chromatography separation and purification product is 0.4 mm-0.5 mm. The column chromatography generally uses 200-300 mesh silica gel of the Tibet Huanghai silica gel as a carrier.
The starting materials in the examples of the present invention are known and commercially available, or may be synthesized using or according to methods known in the art.
All reactions of the present invention are carried out under continuous magnetic stirring in a dry nitrogen or argon atmosphere, without specific indication, the solvent is a dry solvent, and the reaction temperature is given in degrees celsius.
Example 1
Preparation of (S) -1'- (6-amino-5- ((2-amino-3-cyclopropylpyridin-4-yl) thio) pyrazin-2-yl) -1, 3-dihydrospiro [ indene-2, 4' -piperidin ] -1-amine
Figure BDA0003005435270000561
The first step is as follows: preparation of 2-ethylhexyl 3- ((2-amino-3-chloropyridin-4-yl) thio) propionate
Figure BDA0003005435270000562
3-chloro-4-iodopyridin-2-amine (800mg, 3.15mmol), 2-ethylhexyl 3-mercaptopropionate (687mg, 3.15mmol) were dissolved in 1, 4-dioxane (7mL), placed in a microwave reaction tube, DIPEA (813mg, 6.3mmol), palladium acetate (35mg, 0.16mmol) and Xantphos (109mg, 0.19mmol) were added, and after bubbling nitrogen for 3 minutes, the reaction tube was heated to 100 ℃ by microwave for 1 hour. After the reaction solution was cooled, the reaction solution was diluted with 20mL of ethyl acetate, the insoluble material was removed by filtration with celite, and the filtrate was purified by column chromatography (15-25% ethyl acetate/petroleum ether) after spin-drying to give a brown solid product (1.03g, 95% yield).
MS m/z(ESI):345.1[M+H]+.
The second step is that: preparation of 2-ethyl hexyl 3- ((2-amino-3-cyclopropylpyridin-4-yl) thio) propionate
Figure BDA0003005435270000563
Reacting 3- ((2-amino-3-chloro)Pyridin-4-yl) thio) propionic acid (2-ethyl) hexyl ester (500mg, 1.45mmol) was dissolved in a mixed solvent of toluene and water (toluene/water ═ 10mL/1mL), placed in a sealed tube, added with potassium cyclopropyltrifluoroborate (280mg, 1.89mmol), n-butylbis (1-adamantyl) phosphine (52mg, 0.145mmol), palladium acetate (16mg, 0.073mmol) and cesium carbonate (1.41g, 4.35mmol), bubbled with nitrogen for 3 minutes, and then heated to 100 ℃ for 5 hours. After the reaction solution was cooled to room temperature, 20mL of saturated NH was added to the reaction solution4Cl solution, extracted with ethyl acetate (20 mL. times.3); the ethyl acetate layer was washed with a saturated sodium chloride solution, dried over anhydrous sodium sulfate, and purified by column chromatography (50% ethyl acetate/petroleum ether) to give a brown oily product (220mg, yield 43%).
MS m/z(ESI):350.1[M+H]+.
The third step: preparation of potassium 2-amino-3-cyclopropylpyridine-4-mercaptide
Figure BDA0003005435270000571
2-Ethyl-hexyl 3- ((2-amino-3-cyclopropylpyridin-4-yl) thio) propionate (220mg, 0.63mmol) was dissolved in 10mL of ethanol, potassium tert-butoxide (74mg, 0.66mmol) was added, and the mixture was stirred at room temperature for 1 hour. The reaction solution was concentrated to dryness and used directly for the next reaction.
MS m/z(ESI):165.1[M-H]-.
The fourth step: preparation of 3- ((2-amino-3-cyclopropylpyridin-4-yl) thio) -6-chloropyrazin-2-amine
Figure BDA0003005435270000572
2-amino-3-cyclopropylpyridine-4-carbothioic acid potassium (128mg, 0.63mmol), 2-amino-3-bromo-6-chloropyrazine (149mg, 0.72mmol) were dissolved in 7mL of 1, 4-dioxane and placed in a microwave reaction tube, followed by addition of tris (dibenzylideneacetone) dipalladium (33mg, 0.036mmol), Xantphos (42mg, 0.072mmol) and DIPEA (279mg, 2.16mmol) and nitrogen bubbling for 3 minutes. Heating the reaction solution to 110 ℃ by microwave to react for 1 hour; after the reaction solution is cooled to room temperature, the reaction solution is diluted by 20mL of ethyl acetate, filtered by using kieselguhr, and then the filtrate is dried by spinning and purified by column chromatography (50-70% of ethyl acetate/petroleum ether), so that a brown solid product (50mg, 24% of yield) is obtained.
MS m/z(ESI):294.1[M+H]+.
The fifth step: preparation of (R) -N- ((S) -1'- (6-amino-5- ((2-amino-3-cyclopropylpyridin-4-yl) thio) pyrazin-2-yl) -1, 3-dihydrospiro [ indene-2, 4' -piperidin ] -1-yl) -2-methylpropane-2-sulfinamide
Figure BDA0003005435270000573
Taking (S) -1- (((R) -tert-butyl sulfinyl) amino) -1, 3-dihydrospiro [ indene-2, 4' -piperidine]Tert-butyl (69mg, 0.17mmol) of (E) -1' -carboxylate was dissolved in 3mL of dichloromethane, and 1mL of trifluoroacetic acid was added thereto to carry out a reaction at room temperature for 1 hour. The reaction mixture was concentrated to dryness, dissolved in 5mL of DMF, and then added with potassium carbonate (352mg, 2.55mmol) and 3- ((2-amino-3-cyclopropylpyridin-4-yl) thio) -6-chloropyrazin-2-amine (50mg, 0.17mmol), and heated to 100 ℃ under nitrogen for 12 hours. After the reaction solution was cooled, 20mL of water was added, and extraction was performed with ethyl acetate (20 mL. times.3); the ethyl acetate layer was washed with a saturated sodium chloride solution (20 mL. times.3), dried over anhydrous sodium sulfate and purified by column chromatography (75-80% ethyl acetate/petroleum ether) to give an oily product (20mg, 21% yield). MS M/z (ESI) 564.1[ M + H ] ]+.
And a sixth step: preparation of (S) -1'- (6-amino-5- ((2-amino-3-cyclopropylpyridin-4-yl) thio) pyrazin-2-yl) -1, 3-dihydrospiro [ indene-2, 4' -piperidin ] -1-amine
Figure BDA0003005435270000581
Mixing (R) -N- ((S) -1'- (6-amino-5- ((2-amino-3-cyclopropylpyridin-4-yl) thio) pyrazin-2-yl) -1, 3-dihydrospiro [ indene-2, 4' -piperidine]-1-yl) -2-methylpropane-2-sulfinamide (20mg, 0.035mmol) was dissolved in 5mL of dichloromethane, and 1mL of 4M HCl in dioxane was added to react at room temperature for 1 hour. The reaction solution is concentrated to be dry,dissolved in 5mL of methanol and then treated with 7M NH3The reaction solution was adjusted to alkaline pH, column chromatographed (0-10% MeOH in DCM) and the crude product was purified by thin layer chromatography (dichloromethane: methanol 10: 1) to give brown solid product (9.0mg, 56% yield).
1H NMR(400MHz,Methanol-d4)δ7.90(d,J＝5.4Hz,1H),7.61(s,1H),7.40–7.33(m,1H),7.27–7.16(m,3H),6.57(d,J＝5.4Hz,1H),4.28(d,J＝13.6Hz,2H),3.96(s,1H),3.30–3.19(m,2H),3.15(d,J＝15.7Hz,1H),2.81(d,J＝15.7Hz,1H),1.90–1.78(m,2H),1.82–1.70(m,1H),1.58(d,J＝13.5Hz,1H),1.43(d,J＝13.1Hz,1H),1.26–1.15(m,2H),0.94–0.85(m,2H).
MS m/z(ESI):460.1[M+H]+.
Example 2
Preparation of (S) -1'- (6-amino-5- ((2-chloro-3-cyclopropylpyridin-4-yl) thio) pyrazin-2-yl) -1, 3-dihydrospiro [ indene-2, 4' -piperidin ] -1-amine
Figure BDA0003005435270000582
Example 2 was prepared according to the experimental protocol of example 1.
1H NMR(400MHz,Methanol-d4)δ7.88(d,J＝5.5Hz,1H),7.60(s,1H),7.38–7.30(m,1H),7.25–7.15(m,3H),6.54(d,J＝5.5Hz,1H),4.26(d,J＝13.7Hz,2H),3.95(s,1H),3.28–3.17(m,2H),3.13(d,J＝15.6Hz,1H),2.79(d,J＝15.6Hz,1H),1.88–1.74(m,2H),1.80–1.70(m,1H),1.56(d,J＝13.7Hz,1H),1.43(d,J＝13.1Hz,1H),1.24–1.13(m,2H),0.93–0.83(m,2H).
MS m/z(ESI):479.1[M+H]+，481.1[M+2+H]+.
Example 3
Preparation of (S) -1'- (6-amino-5- ((3-chloro-2-cyclopropylpyridin-4-yl) thio) pyrazin-2-yl) -1, 3-dihydrospiro [ indene-2, 4' -piperidin ] -1-amine
Figure BDA0003005435270000591
The first step is as follows: preparation of 2-ethyl hexyl 3- ((2, 3-dichloropyridin-4-yl) thio) propionate
Figure BDA0003005435270000592
2, 3-dichloro-4-iodopyridine (1.0g, 3.65mmol), 2-ethylhexyl 3-mercaptopropionate (0.88g, 4.03mmol) were dissolved in 7mL of 1, 4-dioxane, placed in a microwave reaction tube, DIPEA (0.95g, 7.34mmol), palladium acetate (41mg, 0.18mmol) and Xantphos (127mg, 0.22mmol) were added, nitrogen was bubbled for 3 minutes, and then the mixture was heated to 100 ℃ by microwave for reaction for 1 hour. After the reaction was cooled, the reaction was diluted with 20mL of ethyl acetate, filtered through celite to remove insoluble material, and the filtrate was spin-dried and column chromatographed (10-15% ethyl acetate/petroleum ether) to give a brown oil (1.29g, 97% yield).
MS m/z(ESI):364.1[M+H]+.
The second step is that: preparation of 2-ethyl hexyl 3- ((3-chloro-2-cyclopropylpyridin-4-yl) thio) propionate
Figure BDA0003005435270000593
2-Ethyl hexyl 3- ((2, 3-dichloropyridin-4-yl) thio) propionate (1.29g, 3.54mmol) was dissolved in a mixed solvent of toluene and water (toluene/water 20mL/2mL), placed in a sealed tube, added with potassium cyclopropyltrifluoroborate (624mg, 4.25mmol), n-butylbis (1-adamantyl) phosphine (95mg, 0.267mmol), palladium acetate (40mg, 0.178mmol) and cesium carbonate (3.43g, 3.54mmol), bubbled with nitrogen for 3 minutes, and then heated to 100 ℃ for 5 hours. After the reaction solution was cooled to room temperature, 20mL of saturated NH was added to the reaction solution 4Cl solution, extracted with ethyl acetate (20 mL. times.3); the ethyl acetate layer was washed with a saturated sodium chloride solution, dried over anhydrous sodium sulfate, and subjected to column chromatography (5 to 8% ethyl acetate/petroleum ether) to obtain a yellow oil (600mg, yield 45%).
MS m/z(ESI):370.1[M+H]+.
The third step: preparation of potassium 3-chloro-2-cyclopropylpyridine-4-mercaptide
Figure BDA0003005435270000594
2-ethylhexyl 3- ((3-chloro-2-cyclopropylpyridin-4-yl) thio) propionate (600mg, 1.62mmol) was dissolved in 15mL of ethanol, potassium tert-butoxide (190mg, 1.70mmol) was added, and the mixture was stirred at room temperature for 1 hour. The reaction solution was concentrated to dryness and used directly for the next reaction.
MS m/z(ESI):186.1[M+H]+。
The fourth step: preparation of 6-chloro-3- ((3-chloro-2-cyclopropylpyridin-4-yl) thio) pyrazin-2-amine
Figure BDA0003005435270000601
3-chloro-2-cyclopropylpyridine-4-carbothioic acid potassium (120mg, 0.54mmol), 2-amino-3-bromo-6-chloropyrazine (112mg, 0.54mmol) were dissolved in 5mL of 1, 4-dioxane and placed in a microwave reaction tube, followed by addition of tris (dibenzylideneacetone) dipalladium (25mg, 0.027mmol), Xantphos (31mg, 0.054mmol) and DIPEA (209mg, 1.62mmol) and nitrogen bubbling for 3 minutes. Heating the reaction solution to 110 ℃ by microwave to react for 1 hour; after the reaction solution was cooled to room temperature, it was diluted with 20mL of ethyl acetate, filtered with celite, and the filtrate was subjected to spin-dry column chromatography (10-20% ethyl acetate/petroleum ether) to give an off-white solid product (135mg, yield 80%).
MS m/z(ESI):313.1[M+H]+.
The fifth step: preparation of tert-butyl 1-carbonyl-1, 3-dihydrospiro [ indene-2, 4 '-piperidine ] -1' -carboxylate
Figure BDA0003005435270000602
1-Indenone (5.14g, 38.9mmol), N-bis (2-chloroethyl) carbamic acid tert-butyl ester (9.42g, 38.9mmol) were dissolved in 100mL DMF and 60% sodium hydride (3.89g, 97.3mmol) was added in portions on an ice bath. Will be provided withThe reaction solution is transferred to an oil bath and heated to 60 ℃ under the protection of nitrogen for reaction overnight. After the reaction solution was cooled, 250mL of a saturated sodium chloride solution was added to the reaction solution, followed by extraction with ethyl acetate (150 mL. times.2); the ethyl acetate layer was washed with a saturated sodium chloride solution (100 mL. times.3), anhydrous MgSO4After drying, column chromatography (10-20% ethyl acetate/petroleum ether) afforded crude brown oil (2.80g, 23% yield).
MS m/z(ESI):202.1[M-Boc+H]+.
And a sixth step: preparation of tert-butyl (R, E) -1- ((tert-butylsulfinyl) imino) -1, 3-dihydrospiro [ indene-2, 4 '-piperidine ] -1' -carboxylate
Figure BDA0003005435270000603
Tetraethyl titanate (40mL) was heated to 90 ℃ and tert-butyl 1-carbonyl-1, 3-dihydrospiro [ indene-2, 4 '-piperidine ] -1' -carboxylate (2.80g, 9.27mmol), (R) - (+) -tert-butylsulfinamide (3.36g, 27.8mmol) was added; the reaction is carried out for 24 hours at 90 ℃ under the protection of nitrogen. After the reaction was completed, the reaction solution was poured into 400mL of ethyl acetate, and 400mL of a saturated sodium chloride solution was slowly added with stirring, and stirred at room temperature for 20 minutes. The precipitated solid was filtered with celite, and the obtained filtrate was separated into layers, and the ethyl acetate layer was dried over anhydrous magnesium sulfate and subjected to column chromatography (20-30% ethyl acetate/petroleum ether) to obtain a brown oily product (2.20g, yield 59%).
MS m/z(ESI):405.1[M+H]+.
The seventh step: preparation of (S) -1- (((R) -tert-butylsulfinyl) amino) -1, 3-dihydrospiro [ indene-2, 4 '-piperidine ] -1' -carboxylic acid tert-butyl ester
Figure BDA0003005435270000611
Tert-butyl (R, E) -1- ((tert-butylsulfinyl) imino) -1, 3-dihydrospiro [ indene-2, 4 '-piperidine ] -1' -carboxylate (2.20g, 5.44mmol) was dissolved in 30mL of tetrahydrofuran, placed at-78 ℃ and sodium borohydride (308mg, 8.16mmol) was added portionwise at this temperature and the reaction was allowed to warm gradually to room temperature under nitrogen and stirred overnight. To the reaction solution was added 200mL of a saturated sodium chloride solution, extraction was performed with ethyl acetate (100mL × 2), the ethyl acetate layer was washed with a saturated sodium chloride solution (100mL), dried over anhydrous sodium sulfate, and then column chromatography was performed (20 to 35% ethyl acetate/petroleum ether) to obtain a brown foamy solid (1.21g, yield 54%).
MS m/z(ESI):407.1[M+H]+.
Eighth step: preparation of (R) -N- ((S) -1'- (6-amino-5- ((3-chloro-2-cyclopropylpyridin-4-yl) thio) pyrazin-2-yl) -1, 3-dihydrospiro [ indene-2, 4' -piperidin ] -1-yl) -2-methylpropane-2-sulfinamide
Figure BDA0003005435270000612
(S) -1- (((R) -tert-butylsulfinyl) amino) -1, 3-dihydrospiro [ indene-2, 4 '-piperidine ] -1' -carboxylic acid tert-butyl ester (100mg, 0.25mmol) was dissolved in 3mL of dichloromethane, and 1mL of trifluoroacetic acid was added to react at room temperature for 1 hour. The reaction solution was concentrated to dryness, dissolved in 5mL of DMF, added with potassium carbonate (517mg, 3.75mmol) and 6-chloro-3- ((3-chloro-2-cyclopropylpyridin-4-yl) thio) pyrazin-2-amine (78mg, 0.25mmol), and heated to 100 ℃ under nitrogen protection for 12 hours. After the reaction solution was cooled, 20mL of water was added, and extraction was performed with ethyl acetate (20 mL. times.3); the ethyl acetate layer was washed with a saturated sodium chloride solution (20 mL. times.3), dried over anhydrous sodium sulfate and subjected to column chromatography (50% ethyl acetate/petroleum ether) to give an oily product (26mg, yield 18%).
MS m/z(ESI):583.1[M+H]+.
The ninth step: preparation of (S) -1'- (6-amino-5- ((3-chloro-2-cyclopropylpyridin-4-yl) thio) pyrazin-2-yl) -1, 3-dihydrospiro [ indene-2, 4' -piperidin ] -1-amine
Figure BDA0003005435270000621
Mixing (R) -N- ((S) -1'- (6-amino-5- ((3-chloro-2-cyclopropylpyridin-4-yl) thio) pyrazin-2-yl) -1, 3-dihydrospiro [ indene-2, 4' -piperidine]-1-yl) -2-methylpropane-2-sulfinamide (26mg,0.044mmol) was dissolved in 5mL of dichloromethane, and 1mL of 4M HCl in dioxane was added to react at room temperature for 1 hour. The reaction mixture was concentrated to dryness, dissolved in 5mL of methanol, and then dissolved in 7M NH3Adjusting the pH of the reaction solution to be alkaline, and performing spin-drying and column chromatography (5-8% MeOH in DCM) to obtain a light yellow solid product (12mg, 57% yield).
1H NMR(400MHz,Chloroform-d)δ8.03(d,J＝5.2Hz,1H),7.67(s,1H),7.44–7.37(m,1H),7.24(dd,J＝6.4,3.9Hz,3H),6.39(d,J＝5.2Hz,1H),4.85(s,2H),4.21–4.13(m,2H),4.07(s,1H),3.23–3.21(m,2H),3.13(d,J＝15.8Hz,1H),2.79(d,J＝15.7Hz,1H),2.57–2.48(m,1H),1.88–1.73(m,2H),1.63–1.56(m,1H),1.50–1.42(m,1H),1.09–1.00(m,4H).
MS m/z(ESI):479.1[M+H]+.
Example 4
Preparation of (2R) -1'- (6-amino-5- ((2-amino-3-cyclopropylpyridin-4-yl) thio) pyrazin-2-yl) spiro [ bicyclo [3.1.0] hexane-3, 4' -piperidin ] -2-amine
Figure BDA0003005435270000622
1H NMR(400MHz,Methanol-d4)δ7.73–7.67(m,1H),7.65–7.60(m,1H),6.08–6.01(m,1H),4.53–4.41(m,2H),3.63–3.51(m,2H),3.14(p,J＝9.6Hz,1H),2.41–2.33(m,1H),1.56–1.40(m,3H),1.44–1.35(m,1H),1.39–1.19(m,4H),0.89–0.81(m,2H),0.76–0.60(m,2H),0.44–0.28(m,2H).
MS m/z(ESI):424.1[M+H]+.
Example 4 was prepared according to the experimental protocol of example 1.
Example 5
Preparation of (S) -1'- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) -3-bromopyrazin-2-yl) -1, 3-dihydrospiro [ indene-2, 4' -piperidine ] -1-amine
Figure BDA0003005435270000623
The first step is as follows: preparation of 2-ethyl hexyl 3- ((3-amino-5-chloropyrazin-2-yl) thio) propionate
Figure BDA0003005435270000631
3-bromo-6-chloropyrazin-2-amine (4g,20mmol), 2-ethyl-hexyl 3-mercaptopropionate (5.2g,24mmol), tris (dibenzylideneacetone) dipalladium (916mg,1mmol),4, 5-bis (diphenylphosphino) -9, 9-dimethylxanthene (1.16g,2mmol) and N, N-diisopropylethylamine (5.12g,40mmol) were stirred in dioxane (35mL) at 100 ℃ for 18 hours. The reaction solution was filtered, the filter cake was washed twice with ethyl acetate (30mL), the filtrate was concentrated and purified by column chromatography [ eluent: petroleum ether-petroleum ether/ethyl acetate (90:10) ] to give 2-ethyl-hexyl 3- ((3-amino-5-chloropyrazin-2-yl) thio) propionate (5.5g, yield 82%) as a brown oil.
MS m/z(ESI):146.1[M+H]+，148.1[M+2+H]+.
The second step is that: preparation of 3-amino-5-chloropyrazine-2-thiol
Figure BDA0003005435270000632
Add potassium tert-butoxide (2.7g,23.9mmol) to a solution of 2-ethylhexyl 3- ((3-amino-5-chloropyrazin-2-yl) thio) propionate (5.5g,15.9mmol) in EtOH (100mL) and stir at room temperature for 3 hours. About 50mL of ethanol was spun off, the remaining reaction solution was poured into aqueous ammonium chloride (100mL), extracted twice with ethyl acetate (100mL), extracted twice with dichloromethane (100mL), the organic phase was concentrated, and then purified by column chromatography [ eluent: dichloromethane-dichloromethane/methanol (95:5) ] to give 3-amino-5-chloropyrazine-2-thiol (1.8g, yield 70%) as a dark green solid.
MS m/z(ESI):162.0[M+H]+，164.0[M+2+H]+
The third step: preparation of 3- ((2-amino-3-chloropyridin-4-yl) thio) -6-chloropyrazin-2-amine
Figure BDA0003005435270000633
3-amino-5-chloropyrazine-2-thiol (500mg,3.1mmol), 3-chloro-4-iodopyridin-2-amine (789mg,3.1mmol), tris (dibenzylideneacetone) dipalladium (142mg,0.16mmol), 4, 5-bis (diphenylphosphino) -9, 9-dimethylxanthene (179mg,0.31mmol) and N, N-diisopropylethylamine (1.2g,9.3mmol) were stirred in dioxane (10mL) with microwaves at 130 ℃ for 1 hour. The reaction solution was concentrated and purified by column chromatography [ eluent: dichloromethane to dichloromethane/methanol (99:1) ] to obtain 1g of crude product. Slurried with ethanol (5mL), filtered to give 3- ((2-amino-3-chloropyridin-4-yl) thio) -6-chloropyrazin-2-amine (580mg, 65% yield) as a gray solid.
MS m/z(ESI):288.0[M+H]+，290.0[M+2+H]+.
The fourth step: preparation of N- ((S) -1, 3-dihydrospiro [ indene-2, 4' -piperidine ] -1-yl) -2-methylpropane-2-sulfinamide
Figure BDA0003005435270000641
To a solution of tert-butyl (1S) -1- ((tert-butylsulfinyl < sulfinyl >) amino) -1, 3-dihydrospiro [ indene-2, 4 '-piperidine ] -1' -carboxylate (150mg,0.37mmol) in dichloromethane (3mL) was added trifluoroacetic acid (1mL) and the mixture was stirred at room temperature for 2 hours. The reaction solution was concentrated to give N- ((S) -1, 3-dihydrospiro [ indene-2, 4' -piperidin ] -1-yl) -2-methylpropane-2-sulfinamide hydrochloride (150mg, yield 100%) as a pale yellow solid.
MS m/z(ESI):307.2[M+H]+.
[α]20 D＝1.773.
The fifth step: preparation of N- ((S) -1'- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) pyrazin-2-yl) -1, 3-dihydrospiro [ indene-2, 4' -piperidin ] -1-yl) -2-methylpropane-2-sulfinamide
Figure BDA0003005435270000642
N- ((S) -1, 3-dihydrospiro [ indene-2, 4' -piperidin ] -1-yl) -2-methylpropane-2-sulfinamide hydrochloride (150mg,0.37mmol), 3- ((2-amino-3-chloropyridin-4-yl) thio) -6-chloropyrazin-2-amine (100mg,0.35mmol) and potassium carbonate (335mg,2.43mmol) were stirred in N, N-dimethylformamide (4mL) at 100 ℃ for 18 h. The reaction solution was concentrated and purified by column chromatography [ eluent: dichloromethane to dichloromethane/methanol (97:3) ] to give N- ((S) -1'- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) pyrazin-2-yl) -1, 3-dihydrospiro [ indene-2, 4' -piperidin ] -1-yl) -2-methylpropane-2-sulfinamide (100mg, yield 52%) as a purple solid.
MS m/z(ESI):558.1[M+H]+，560.2[M+2+H]+.
And a sixth step: preparation of (S) -1'- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) -3-bromopyrazin-2-yl) -1, 3-dihydrospiro [ indene-2, 4' -piperidine ] -1-amine
Figure BDA0003005435270000643
N- ((S) -1' - (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) pyrazin-2-yl) -1, 3-dihydrospiro [ indene-2, 4' -piperidine ] -1-yl) -2-methylpropane-2-sulfinamide (100mg, 0.18mmol) and N-bromosuccinimide (64mg, 0.36mmol) are stirred in N, N-dimethylformamide (1mL) at room temperature for 18 hours, the reaction solution is concentrated to obtain a crude product, and column chromatography [ eluent: dichloromethane-dichloromethane/methanol (95:5) ] is purified to obtain (S) -1' - (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) -3-bromopyrazine -2-yl) -1, 3-dihydrospiro [ indene-2, 4' -piperidine ] -1-amine (80mg, 84% yield) as an earthy yellow solid.
1H NMR(400MHz,DMSO-d6)δ7.72-7.64(m,1H),7.36(d,J＝4Hz,1H),7.27-7.16(m,3H),6.58(s,2H),6.33(s,2H),5.80(d,J＝4Hz,1H),3.99(s,1H),3.94-3.83(m,2H),3.17(d,J＝4Hz,1H),3.14-2.98(m,3H),2.77-2.64(m,1H),1.97-1.74(m,2H),1.55(d,J＝12Hz,1H).
MS m/z(ESI):532.0[M+H]+,534.0[M+2+H]+.
Example 6
Preparation of (S) -1'- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) -3-methylpyrazin-2-yl) -1, 3-dihydrospiro [ indene-2, 4' -piperidine ] -1-amine
Figure BDA0003005435270000651
The first step is as follows: preparation of (S) -1'- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) -3-methylpyrazin-2-yl) -1, 3-dihydrospiro [ indene-2, 4' -piperidine ] -1-amine
Figure BDA0003005435270000652
(S) -1' - (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) -3-bromopyrazin-2-yl) -1, 3-dihydrospiro [ indene-2, 4' -piperidine ] -1-amine (30mg,0.056mmol), trimethylcyclotriboroxane (789mg,3.1mmol), [1,1' -bis (diphenylphosphino) ferrocene ] palladium dichloride (4mg,0.0056mmol), and potassium carbonate (15mg,0.11mmol) were stirred in N, N-dimethylformamide (10mL) with microwave agitation at 130 ℃ for 1 hour. The reaction solution was concentrated and purified by HPLC to give (S) -1'- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) -3-methylpyrazin-2-yl) -1, 3-dihydrospiro [ indene-2, 4' -piperidine ] -1-amine (0.7mg, yield 3%) as a gray solid.
1H NMR(400MHz,DMSO)δ7.73-7.62(m,1H),7.35(d,J＝4Hz,1H),7.28-7.16(m,3H),6.57(s,2H),6.32(s,2H),5.78(d,J＝4Hz,1H),3.97(s,1H),3.94-3.81(m,2H),3.15(d,J＝4Hz,1H),3.12-2.95(m,3H),2.75-2.64(m,1H),2.41(s,3H),1.99-1.75(m,2H),1.58(d,J＝12Hz,1H).
MS m/z(ESI):468.1[M+H]+，470.1[M+2+H]+.
Example 7
Preparation of (S) -1'- (5- ((3-chloro-2-cyclopropylpyridin-4-yl) thio) pyrazin-2-yl) -1, 3-dihydrospiro [ indene-2, 4' -piperidin ] -1-amine
Figure BDA0003005435270000653
The first step is as follows: preparation of 2-chloro-5- ((3-chloro-2-cyclopropylpyridin-4-yl) thio) pyrazine
Figure BDA0003005435270000661
3-chloro-2-cyclopropylpyridine-4-carbothioic acid potassium (120mg, 0.54mmol), 2-bromo-5-chloropyrazine (104mg, 0.54mmol) were dissolved in 5mL dioxane and placed in a microwave reaction tube, followed by addition of tris (dibenzylideneacetone) dipalladium (25mg, 0027mmol), Xantphos (31mg, 0.054mmol) and DIPEA (209mg, 1.62mmol), and nitrogen bubbling for 3 minutes. Heating the reaction solution to 110 ℃ by microwave to react for 1 hour; after the reaction solution was cooled to room temperature, it was diluted with 20mL of ethyl acetate, filtered through celite, and the filtrate was subjected to spin-dry column chromatography (10-20% ethyl acetate/petroleum ether) to give a pale yellow oil (120mg, 74% yield).
MS m/z(ESI):298.0[M+H]+.
The second step is that: preparation of (R) -N- ((S) -1'- (5- ((3-chloro-2-cyclopropylpyridin-4-yl) thio) pyrazin-2-yl) -1, 3-dihydrospiro [ indene-2, 4' -piperidin ] -1-yl) -2-methylpropane-2-sulfinamide
Figure BDA0003005435270000662
Tert-butyl (S) -1- (((R) -tert-butylsulfinyl) amino) -1, 3-dihydrospiro [ indene-2, 4 '-piperidine ] -1' -carboxylate (81mg, 0.20mmol) was dissolved in 3mL of dichloromethane, and 1mL of trifluoroacetic acid was added to react at room temperature for 1 hour. The reaction mixture was concentrated to dryness, dissolved in 5mL of DMF, added with potassium carbonate (373mg, 2.70mmol) and 2-chloro-5- ((3-chloro-2-cyclopropylpyridin-4-yl) thio) pyrazine (55mg, 0.18mmol), and heated to 100 ℃ under nitrogen for 12 hours. After the reaction solution was cooled, 20mL of water was added, and extraction was performed with ethyl acetate (20 mL. times.3); the ethyl acetate layer was washed with a saturated sodium chloride solution (20 mL. times.3), dried over anhydrous sodium sulfate and subjected to column chromatography (50% ethyl acetate/petroleum ether) to give an oily product (48mg, yield 47%).
MS m/z(ESI):568.1[M+H]+.
The third step: preparation of (S) -1'- (5- ((3-chloro-2-cyclopropylpyridin-4-yl) thio) pyrazin-2-yl) -1, 3-dihydrospiro [ indene-2, 4' -piperidin ] -1-amine
Figure BDA0003005435270000663
Mixing (R) -N- ((S) -1'- (5- ((3-chloro-2-cyclopropylpyridin-4-yl) thio) pyrazin-2-yl) -1, 3-dihydrospiro [ indene-2, 4' -piperidine]-1-yl) -2-methylpropane-2-sulfinamide (48mg, 0.085mmol) was dissolved in 5mL of dichloromethane, and 1mL of 4M HCl in dioxane was added and reacted at room temperature for 1 hour. The reaction mixture was concentrated to dryness, dissolved in 5mL of methanol, and then dissolved in 7M NH3Adjusting the pH of the reaction solution to be alkaline, and performing spin-drying and column chromatography (0-10% MeOH in DCM) to obtain a light yellow solid product (23mg, 58% yield).
1H NMR(400MHz,Methanol-d4)δ8.36(s,1H),8.30(s,1H),7.98(d,J＝5.3Hz,1H),7.42(d,J＝6.8Hz,1H),7.32–7.21(m,3H),6.43(d,J＝5.3Hz,1H),4.44–4.29(m,2H),4.14(s,1H),3.41–3.30(m,2H),3.19(d,J＝15.9Hz,1H),2.96(d,J＝15.9Hz,1H),2.60–2.49(m,1H),1.90–1.76(m,2H),1.67–1.54(m,2H),1.09–0.99(m,4H).
MS m/z(ESI):464.1[M+H]+.
Example 8
Preparation of (S) -1'- (6-amino-5- ((3-chloro-2-cyclopropylpyridin-4-yl) thio) pyrazin-2-yl) -4, 6-dihydrospiro [ cyclopenta [ d ] thiazole-5, 4' -piperidine ] -6-amine
Figure BDA0003005435270000671
The first step is as follows: preparation of tert-butyl (5-bromo-1, 3, 4-thiadiazol-2-yl) carbamate
Figure BDA0003005435270000672
Ethyl 2-chlorothiazole-4-carboxylate (9.0g, 46.97mmol) was dissolved in 150mL of methanol, stirred at room temperature, and NaBH added4(7.1g, added in three portions) and stirred until the starting material disappeared. Then theThe reaction solution was slowly added to stirred brine, quenched, and the solvent was evaporated off, extracted three times with ethyl acetate (200mL), the organic layers were combined, dried over anhydrous sodium sulfate, and concentrated to give a crude product, which was purified by column chromatography (ethyl acetate/petroleum ether ═ 3: 1) to give (2-chlorothiazol-4-yl) methanol (4.2g, yield 60%) as a colorless oily liquid target product.
1H NMR(400MHz,CDCl3)δ7.11(s,1H),4.71(s,2H),2.22(s,1H).
MS m/z(ESI):150.0[M+H]+，152.0[M+2+H]+.
The second step is that: preparation of (2-chlorothiazol-4-yl) methyl methanesulfonate
Figure BDA0003005435270000673
(2-chlorothiazol-4-yl) methanol (2.6g, 17.4mmol) was dissolved in 40mL of dichloromethane, and methanesulfonyl chloride (1.6mL,20.88mmol, d ═ 1.48g/mL) was added dropwise under nitrogen protection in an ice-water bath, and the mixture was stirred for 30min under an ice-water bath. At the end of the reaction, the reaction was added dropwise to saturated sodium chloride (100mL), extracted three times with dichloromethane (100mL), the organic layers were combined, dried over anhydrous sodium sulfate, and concentrated to give the title product (2-chlorothiazol-4-yl) methyl methanesulfonate as a white solid (3.9g, 97% yield).
1H NMR(400MHz,CDCl3)δ7.35(s,1H),5.25(s,2H),3.07(s,3H).
The third step: preparation of 1- (tert-butyl) -4-ethyl 4- ((2-chlorothiazol-4-yl) methyl) piperidine-1, 4-dicarboxylate
Figure BDA0003005435270000681
Piperidine-1, 4-dicarboxylic acid 1- (tert-butyl) -4-ethyl ester (4.8g, 18.59mmol) was dissolved in THF (40mL), cooled to-60 ℃ under nitrogen protection, lithium diisopropylamide (12.7mL, 25.35mmol) was added dropwise, and the reaction mixture was stirred at-60 ℃ to-50 ℃ for 30min after completion of the addition. A solution of (2-chlorothiazol-4-yl) methyl methanesulfonate in THF (15mL) was then added dropwise. After the dropwise addition, the mixture was stirred at-60 ℃ for 30min, and then slowly warmed to room temperature and stirred for 2 hours. After completion of the reaction, the reaction solution was added dropwise to saturated brine (150 mL). Ethyl acetate (100mL) was extracted three times. The combined organic layers were dried over anhydrous sodium sulfate, concentrated to give crude product which was purified by column chromatography (ethyl acetate/petroleum ether ═ 3: 1) to give 4- ((2-chlorothiazol-4-yl) methyl) piperidine-1, 4-dicarboxylic acid 1- (tert-butyl) -4-ethyl ester (3.7g, 56% yield) as a yellow liquid.
1H NMR(400MHz,CDCl3)δ6.79(s,1H),4.14(dd,J＝7.1,3.1Hz,2H),3.91–3.85(m,2H),2.92(s,2H),2.86(dd,J＝16.3,13.6Hz,2H),2.10(d,J＝13.5Hz,2H),1.53(s,2H),1.44(s,9H),1.25–1.22(m,3H).
MS m/z(ESI):289[M-100+H]+.
The fourth step: preparation of tert-butyl 2-chloro-6-carbonyl-4, 6-dihydrospiro [ cyclopenta [ d ] thiazole-5, 4 '-piperidine ] -1' -carboxylate
Figure BDA0003005435270000682
4-Ethyl-4- ((2-chlorothiazol-4-yl) methyl) piperidine-1, 4-dicarboxylic acid 1- (tert-butyl) -4-ethyl ester (4.1g, 10.6mmol) was dissolved in THF (40mL), cooled to-70 ℃ under nitrogen protection, lithium diisopropylamide (13.4mL, 26.5mmol) was added dropwise, and the reaction mixture was stirred at-70 ℃ for 30 min. After completion of the reaction, the reaction solution was added dropwise to saturated brine (150mL), and extracted three times with ethyl acetate (100 mL). The organic layers were combined, dried over anhydrous sodium sulfate, concentrated to give the crude product, which was purified by column chromatography (ethyl acetate/petroleum ether ═ 3: 1) to give the desired tert-butyl 2-chloro-6-carbonyl-4, 6-dihydrospiro [ cyclopenta [ d ] thiazole-5, 4 '-piperidine ] -1' -carboxylate (1.2g, 33% yield) as a yellow solid.
1H NMR(400MHz,CDCl3)δ4.15(s,4H),3.05(s,2H),2.06–1.93(m,2H),1.60(d,J＝12.7Hz,2H),1.48(s,9H).
MS m/z(ESI):243[M-100+H]+.
The fifth step: preparation of tert-butyl (R, Z) -6- ((tert-butylsulfinyl < sulfinyl >) imino) -2-chloro-4, 6-dihydrospiro [ cyclopenta [ d ] thiazole-5, 4 '-piperidine ] -1' -carboxylate
Figure BDA0003005435270000691
Tert-butyl 2-chloro-6-carbonyl-4, 6-dihydrospiro [ cyclopenta [ d ]]Thiazole-5, 4' -piperidines]-1' -Carboxylic acid ester (1.2g, 3.5mmol) was dissolved in THF (10mL), Ti (OEt) was added4(10mL) was heated to 95 ℃ under nitrogen and stirred for 10 hours. After cooling to room temperature, the reaction mixture was diluted with ethyl acetate (200mL), water (20mL) was added with stirring, and stirring was continued until the mixture became turbid. Filtering, concentrating the filtrate to obtain a crude product, and purifying by column chromatography (PE/EA is 5: 1) to obtain a yellow solid target product (R, Z) -6- ((tert-butyl sulfinyl) <Sulfinyl radical>) Imino) -2-chloro-4, 6-dihydrospiro [ cyclopenta [ d ]]Thiazole-5, 4' -piperidines]-1' -Carboxylic acid tert-butyl ester (1.1g, yield 75%).
1H NMR(400MHz,CDCl3)δ4.13(dd,J＝22.7,15.6Hz,4H),2.92(d,J＝2.9Hz,2H),2.05(s,2H),1.57(s,2H),1.48(s,9H),1.29–1.22(m,9H).
MS m/z(ESI):346[M-100+H]+.
And a sixth step: (S) -6- (((R) -tert-butylsulfinyl group<Sulfinyl radical>) Amino) -4, 6-dihydrospiro [ cyclopenta [ d ]]Thiazole-5, 4' -piperidines]-1' -Carboxylic acid ester and tert-butyl (S) -6- (((R) -tert-butylsulfinyl)<Sulfinyl radical>) Amino) -2-chloro-4, 6-dihydrospiro [ cyclopenta [ d ]]Thiazole-5, 4' -piperidines]Preparation of tert-butyl (E) -1' -carboxylate
Figure BDA0003005435270000692
Reacting (R, Z) -6- ((tert-butylsulfinyl)<Sulfinyl radical>) Imino) -2-chloro-4, 6-dihydrospiro [ cyclopenta [ d ]]Thiazole-5, 4' -piperidines]-1' -Carboxylic acid tert-butyl ester (600mg, 1.35mmol) is dissolved in anhydrous THF (10mL), placed in a 50mL three-necked flask, cooled to-70 ℃ under nitrogen, and B is added dropwiseH3THF (4.0mL, D ═ 1M), added dropwise, slowly warmed to room temperature and stirred for 10 hours. The reaction was added dropwise to water, saturated with sodium bicarbonate to pH 9, and extracted three times with ethyl acetate (100 mL). Combining organic layers, drying by anhydrous sodium sulfate, concentrating to obtain a crude product, and purifying by column chromatography (PE/EA is 1: 1) to obtain a yellow liquid target product, namely tert-butyl (S) -6- (((R) -tert-butyl sulfinyl)<Sulfinyl radical>) Amino) -4, 6-dihydrospiro [ cyclopenta [ d ] ]Thiazole-5, 4' -piperidines]-1' -Carboxylic acid ester (112mg, yield 20%) and yellow liquid (S) -6- (((R) -tert-butylsulfinyl)<Sulfinyl radical>) Amino) -2-chloro-4, 6-dihydrospiro [ cyclopenta [ d ]]Thiazole-5, 4' -piperidines]Tert-butyl (56mg, yield 9%) 1' -carboxylate.
MS m/z(ESI):414.0[M+H]+；
MS m/z(ESI):448.1[M+H]+，450.1[M+H+2]+.
The seventh step: preparation of (R) -N- ((S) -4, 6-dihydrospiro [ cyclopenta [ d ] thiazole-5, 4' -piperidin ] -6-yl) -2-methylpropane-2-sulfinamide
Figure BDA0003005435270000701
Reacting (S) -6- (((R) -tert-butylsulfinyl)<Sulfinyl radical>) Amino) -4, 6-dihydrospiro [ cyclopenta [ d ]]Thiazole-5, 4' -piperidines]-1' -Carboxylic acid tert-butyl ester (112mg, 0.27mmol) dissolved in anhydrous CH2Cl2(5mL), trifluoroacetic acid (1.0mL) was added thereto, and the mixture was stirred at room temperature for 2 hours, followed by concentration of the reaction mixture to give a crude (R) -N- ((S) -4, 6-dihydrospiro [ cyclopenta [ d ] as a yellow liquid]Thiazole-5, 4' -piperidines]-6-yl) -2-methylpropane-2-sulfinamide (67mg, 76% yield).
MS m/z(ESI):314.0[M+H]+.
Eighth step: preparation of (R) -N- ((S) -1'- (6-amino-5- ((3-chloro-2-cyclopropylpyridin-4-yl) thio) pyrazin-2-yl) -4, 6-dihydrospiro [ cyclopenta [ d ] thiazole-5, 4' -piperidin ] -6-yl) -2-methylpropane-2-sulfinamide
Figure BDA0003005435270000702
(R) -N- ((S) -4, 6-dihydrospiro [ cyclopenta [ d ] thiazole-5, 4' -piperidin ] -6-yl) -2-methylpropane-2-sulfinamide (67mg, 0.21mmol), 6-chloro-3- ((3-chloro-2-cyclopropylpyridin-4-yl) thio) pyrazin-2-amine (78mg,0.25mmol) and DIPEA (0.4mL) were dissolved in DMF (2.0mL), heated to 110 ℃ and stirred for 10 hours. The reaction mixture was dissolved in ethyl acetate (100mL), washed twice with water (50mL) and washed three times with saturated brine (50 mL). The combined organic layers were dried over anhydrous sodium sulfate and concentrated to give the crude product which was purified by column chromatography (EtOAc) to give (R) -N- ((S) -1'- (6-amino-5- ((3-chloro-2-cyclopropylpyridin-4-yl) thio) pyrazin-2-yl) -4, 6-dihydrospiro [ cyclopenta [ d ] thiazole-5, 4' -piperidin ] -6-yl) -2-methylpropane-2-sulfinamide (30mg, 19% yield) as a yellow liquid target.
MS m/z(ESI):590.0[M+H]+.
The ninth step: preparation of (S) -1'- (6-amino-5- ((3-chloro-2-cyclopropylpyridin-4-yl) thio) pyrazin-2-yl) -4, 6-dihydrospiro [ cyclopenta [ d ] thiazole-5, 4' -piperidine ] -6-amine
Figure BDA0003005435270000703
(R) -N- ((S) -1'- (6-amino-5- ((3-chloro-2-cyclopropylpyridin-4-yl) thio) pyrazin-2-yl) -4, 6-dihydrospiro [ cyclopenta [ d ] thiazole-5, 4' -piperidin ] -6-yl) -2-methylpropane-2-sulfinamide (30mg, 0.05mmol) was dissolved in anhydrous methanol (3.0mL), hydrochloric acid/1, 4-dioxane (1.0mL) was added, stirring was carried out at room temperature for 0.5 hour, and the reaction solution was concentrated to give a crude product, which was dissolved in water, adjusted to pH 10 with a saturated solution of sodium hydrogencarbonate, and extracted twice with dichloromethane (50 mL). The organic layers were combined, dried over anhydrous sodium sulfate, concentrated to give the crude product, which was purified by preparative HPLC to afford the target (S) -1'- (6-amino-5- ((3-chloro-2-cyclopropylpyridin-4-yl) thio) pyrazin-2-yl) -4, 6-dihydrospiro [ cyclopenta [ d ] thiazole-5, 4' -piperidine ] -6-amine as a yellow solid (3.4mg, 49% yield).
1H NMR(500MHz,CDCl3)δ8.77(s,1H),8.48(s,1H),7.45(d,J＝9.7Hz,2H),4.45(s,1H),3.38(d,J＝61.4Hz,4H),2.91(s,2H),2.22(s,1H),1.92(s,1H),1.71(s,1H),1.52(s,2H),1.33–0.92(m,8H).
MS m/z(ESI):486[M+H]+.
Example 9
Preparation of (S) -1'- (6-amino-5- ((3-chloro-2-cyclopropylpyridin-4-yl) thio) pyrazin-2-yl) -2-chloro-4, 6-dihydrospiro [ cyclopenta [ d ] thiazole-5, 4' -piperidin ] -6-amine
Figure BDA0003005435270000711
The first step is as follows: preparation of (R) -N- ((S) -2-chloro-4, 6-dihydrospiro [ cyclopenta [ d ] thiazole-5, 4' -piperidin ] -6-yl) -2-methylpropane-2-sulfinamide
Figure BDA0003005435270000712
Reacting (S) -6- (((R) -tert-butylsulfinyl)<Sulfinyl radical>) Amino) -2-chloro-4, 6-dihydrospiro [ cyclopenta [ d ]]Thiazole-5, 4' -piperidines]-1' -Carboxylic acid tert-butyl ester (56mg, 0.13mmol) dissolved in anhydrous CH2Cl2(5mL), trifluoroacetic acid (1.0mL) was added thereto, and the mixture was stirred at room temperature for 2 hours, followed by concentration of the reaction mixture to give a crude (R) -N- ((S) -2-chloro-4, 6-dihydrospiro [ cyclopenta [ d ] as a yellow liquid]Thiazole-5, 4' -piperidines]-6-yl) -2-methylpropane-2-sulfinamide (40mg, 100% yield).
MS m/z(ESI):348.0[M+H]+.
The second step is that: (R) -N- ((S) -1' - (6-amino-5- ((3-chloro-2-cyclopropylpyridin-4-yl) thio) pyrazin-2-yl) -2-chloro-4, 6-dihydrospiro [ cyclopenta [ d)]Thiazole-5, 4' -piperidines]Preparation of (E) -6-yl-2-methylpropane-2-sulfinamide
Figure BDA0003005435270000713
(R) -N- ((S) -2-chloro-4, 6-dihydrospiro [ cyclopenta [ d ] thiazol-5, 4' -piperidin ] -6-yl) -2-methylpropane-2-sulfinamide (40mg, 0.1mmol), 6-chloro-3- ((3-chloro-2-cyclopropylpyridin-4-yl) thio) pyrazin-2-amine (15mg,0.05mmol) and DIPEA (0.1mL) were dissolved in DMF (0.8mL), heated to 110 ℃ and stirred for 10 hours. The reaction mixture was dissolved in ethyl acetate (100mL), washed with water (50mL) twice, and saturated brine (50mL) three times. The combined organic layers were dried over anhydrous sodium sulfate and concentrated to give the crude product which was purified by column chromatography (EtOAc) to give (R) -N- ((S) -1'- (6-amino-5- ((3-chloro-2-cyclopropylpyridin-4-yl) thio) pyrazin-2-yl) -4, 6-dihydrospiro [ cyclopenta [ d ] thiazole-5, 4' -piperidin ] -6-yl) -2-methylpropane-2-sulfinamide (8.2mg, 26% yield) as a yellow liquid target.
MS m/z(ESI):624.1[M+H]+.
The third step: preparation of (S) -1'- (6-amino-5- ((3-chloro-2-cyclopropylpyridin-4-yl) thio) pyrazin-2-yl) -2-chloro-4, 6-dihydrospiro [ cyclopenta [ d ] thiazole-5, 4' -piperidin ] -6-amine
Figure BDA0003005435270000721
Dissolving (R) -N- ((S) -1' - (6-amino-5- ((3-chloro-2-cyclopropylpyridin-4-yl) thio) pyrazin-2-yl) -4, 6-dihydrospiro [ cyclopenta [ d ] thiazole-5, 4' -piperidin ] -6-yl) -2-methylpropane-2-sulfinamide (8.2mg, 0.01mmol) in anhydrous methanol (2.0mL), adding hydrochloric acid/dioxane (0.5mL), stirring at room temperature for 0.5 hour, concentrating the reaction solution to obtain a crude product, dissolving in water, adjusting the pH to 10 with a saturated solution of sodium bicarbonate, extracting twice with dichloromethane (50mL), combining the organic layers, drying over anhydrous sodium sulfate, concentrating to obtain a crude product, and purifying by preparative HPLC to obtain a yellow solid target product (S) -1' - (6-amino-substituted) 5- ((3-chloro-2-cyclopropylpyridin-4-yl) thio) pyrazin-2-yl) -2-chloro-4, 6-dihydrospiro [ cyclopenta [ d ] thiazole-5, 4' -piperidine ] -6-amine (1.5mg, 22% yield).
1H NMR(500MHz,CDCl3)δ8.48(s,1H),7.45(d,J＝10.3Hz,2H),4.71(s,1H),3.41(d,J＝74.5Hz,4H),2.92(s,2H),2.16(d,J＝56.0Hz,2H),1.74(s,1H),1.52(s,2H),1.26(d,J＝15.5Hz,4H),1.03(d,J＝39.0Hz,4H).
MS m/z(ESI):520.1[M+H]+.
Example 10
Preparation of (S) -1'- (6-amino-5- ((2- (azetidin-1-yl) -3-chloropyridin-4-yl) thio) pyrazin-2-yl) -1, 3-dihydrospiro [ indene-2, 4' -piperidine ] -1-amine
Figure BDA0003005435270000722
The first step is as follows: preparation of 2-ethylhexyl 3- ((2, 3-dichloropyridin-4-yl) thio) propionate
Figure BDA0003005435270000731
2, 3-dichloro-4-iodopyridine (2.0g, 7.31mmol), 2-ethylhexyl 3-mercaptopropionate (1.8g, 8.24mmol), palladium acetate (82mg, 0.37mmol), Xantphos (254mg, 0.44mmol), diisopropylethylamine (1.9g, 14.70mmol) in 15mL dioxane was stirred at 100 ℃ under nitrogen for 5h, cooled to room temperature, added with ethyl acetate, filtered, concentrated to dryness, and purified by column chromatography (petroleum ether/ethyl acetate ═ 3: 1) to give the desired product, 2-ethylhexyl 3- ((2, 3-dichloropyridin-4-yl) thio) propionate (910mg, 34% yield).
MS m/z(ESI):364.1[M+H]+
The second step is that: preparation of 2-ethylhexyl 3- ((2- (azetidin-1-yl) -3-chloropyridin-4-yl) thio) propionate
Figure BDA0003005435270000732
2-ethylhexyl 3- ((2, 3-dichloropyridin-4-yl) thio) propionate (200mg, 0.55mmol), azetidine (63mg, 1.10mmol), 1, 4-dioxane (5mL) was stirred with a microwave at 120 ℃ for 3 h. Concentrate to dryness, extract with water, dichloromethane, concentrate to dryness, and column chromatographe (3: 1 petroleum ether/ethyl acetate) to give the product (130mg, 62% yield).
MS m/z(ESI):385.1[M+H]+.
The third step: preparation of 2- (azetidin-1-yl) -3-chloropyridine-4-thiol
Figure BDA0003005435270000733
2-ethylhexyl 3- ((2- (azetidin-1-yl) -3-chloropyridin-4-yl) thio) propanoate (130mg, 0.34mmol), potassium tert-butoxide (38mg, 0.34mmol) in absolute ethanol (5mL) was stirred at room temperature for 1h and concentrated to dryness to give the product which was used directly in the next reaction.
MS m/z(ESI):201.0[M+H]+.
The fourth step: preparation of 3- ((2- (azetidin-1-yl) -3-chloropyridin-4-yl) thio) -6-chloropyrazin-2-amine
Figure BDA0003005435270000734
2- (azetidin-1-yl) -3-chloropyridine-4-thiol (from the previous step), 3-bromo-6-chloropyrazin-2-amine (84mg, 0.40mmol), Pd2(dba)3(15mg, 0.016mmol), Xantphos (20mg, 0.035mmol), diisopropylethylamine (87mg, 0.67mmol) was added to 5mL of 1, 4-dioxane, stirred overnight at 100 ℃ under nitrogen, cooled to room temperature, concentrated to dryness, and purified by column chromatography (petroleum ether/ethyl acetate ═ 1: 1) to give a pale yellow solid (71mg, 64% yield).
MS m/z(ESI):328.0[M+H]+.
The fifth step: preparation of (R) -N- ((S) -1, 3-dihydrospiro [ indene-2, 4' -piperidine ] -1-yl) -2-methylpropane-2-sulfinamide
Figure BDA0003005435270000741
(S) -1- (((R) -tert-butylsulfinyl < sulfinyl >) amino) -1, 3-dihydrospiro [ indene-2, 4 '-piperidine ] -1' -carboxylic acid tert-butyl ester (100mg, 0.25mmol) was dissolved in dichloromethane (1mL), 1mL trifluoroacetic acid was added, stirred at room temperature for 1h, and concentrated to dryness to give an oil which was used directly in the next reaction.
MS m/z(ESI):307.1[M+H]+.
[α]20 D＝1.773.
And a sixth step: preparation of (R) -N- ((S) -1'- (6-amino-5- ((2- (azetidin-1-yl) -3-chloropyridin-4-yl) thio) pyrazin-2-yl) -1, 3-dihydrospiro [ indene-2, 4' -piperidin ] -1-yl) -2-methylpropane-2-sulfinamide
Figure BDA0003005435270000742
3- ((2- (azetidin-1-yl) -3-chloropyridin-4-yl) thio) -6-chloropyrazin-2-amine (71mg, 0.22mmol), (R) -N- ((S) -1, 3-dihydrospiro [ indene-2, 4' -piperidin ] -1-yl) -2-methylpropane-2-sulfinamide (obtained in the previous step), potassium carbonate (299mg, 2.16mmol) in DMF (5mL) under nitrogen at 100 ℃ with microwave stirring for 2h, water was added and ethyl acetate was extracted. Dried over anhydrous sodium sulfate and concentrated to dryness to give an oil which was used directly in the next reaction.
MS m/z(ESI):598.2[M+H]+.
The seventh step: preparation of (S) -1'- (6-amino-5- ((2- (azetidin-1-yl) -3-chloropyridin-4-yl) thio) pyrazin-2-yl) -1, 3-dihydrospiro [ indene-2, 4' -piperidine ] -1-amine
Figure BDA0003005435270000743
(R) -N- ((S) -1' - (6-amino-5- ((2- (azetidin-1-yl) -3-chloropyridin-4-yl) thio) pyrazin-2-yl) -1, 3-dihydrospiro [ indene-2, 4' -piperidine ] -1-yl) -2-methylpropane-2-sulfinamide (obtained in the previous step) is dissolved in methanol (5mL), 4M HCl/1, 4-dioxane (5mL) is added and stirred for 1h at room temperature, the mixture is concentrated to dryness, and the target product (S) -1' - (6-amino-5- ((2- (azetidin-1-yl) -3-chloropyridin-4-yl) thio) pyrazin-2-yl) is obtained after column chromatography purification 1, 3-dihydrospiro [ indene-2, 4' -piperidine ] -1-amine (8.7mg, yield 8%).
1H NMR(400MHz,Methanol-d4)δ7.69(d,J＝5.4Hz,1H),7.58(s,1H),7.42–7.32(m,1H),7.28–7.14(m,3H),5.97(d,J＝5.5Hz,1H),4.35–4.16(m,6H),3.96(s,1H),3.28–3.19(m,2H),3.15(d,J＝15.7Hz,1H),2.81(d,J＝15.7Hz,1H),2.32(q,J＝7.5Hz,2H),1.80(dtd,J＝32.3,12.6,4.3Hz,2H),1.58(d,J＝13.4Hz,1H),1.42(d,J＝13.4Hz,1H),1.31(d,J＝16.2Hz,1H).
MS m/z(ESI):494.1[M+H]+.
Example 11
Preparation of (S) -1'- (6-amino-5- ((3-chloro-2-morpholinopyridin-4-yl) thio) pyrazin-2-yl) -1, 3-dihydrospiro [ indene-2, 4' -piperidin ] -1-amine
Figure BDA0003005435270000751
The first step is as follows: preparation of 2-ethyl hexyl 3- ((3-chloro-2-morpholinopyridin-4-yl) thio) propionate
Figure BDA0003005435270000752
2-Ethyl-hexyl 3- ((2, 3-dichloropyridin-4-yl) thio) propionate (200mg, 0.55mmol), morpholine (96mg, 1.10mmol) and 1, 4-dioxane (5mL) were stirred with a microwave at 120 ℃ for 8 h. Concentrating to dryness, adding water, and extracting with dichloromethane. Concentrate to dryness and column chromatographe (3: 1 petroleum ether/ethyl acetate) to give the product (152mg, 67% yield).
MS m/z(ESI):415.2[M+H]+.
The second step is that: preparation of potassium 3-chloro-2-morpholinopyridine-4-mercaptide
Figure BDA0003005435270000753
2-Ethyl-hexyl 3- ((3-chloro-2-morpholinopyridin-4-yl) thio) propionate (100mg, 0.24mmol), potassium tert-butoxide (27mg, 0.24mmol) in absolute ethanol (5mL) was stirred at room temperature for 1 h. The product was obtained by concentration to dryness and used directly in the next reaction.
MS m/z(ESI):231.0[M+H]+.
The third step: preparation of 6-chloro-3- ((3-chloro-2-morpholinopyridin-4-yl) thio) pyrazin-2-amine
Figure BDA0003005435270000761
Potassium 3-chloro-2-morpholinopyridin-4-thiolate (obtained in the previous step), 3-bromo-6-chloropyrazin-2-amine (60mg, 0.29mmol), and Pd2(dba)3(11mg, 0.012mmol), Xantphos (14mg, 0.024mmol) and diisopropylethylamine (62mg, 0.48mmol) in 5mL dioxane, stirring overnight at 100 ℃ under nitrogen. Cool to room temperature, concentrate to dryness, and purify by column chromatography (petroleum ether/ethyl acetate ═ 1: 1) to give a pale yellow solid (60mg, 70% yield).
MS m/z(ESI):358.0[M+H]+.
The fourth step: preparation of (R) -N- ((S) -1'- (6-amino-5- ((3-chloro-2-morpholinopyridin-4-yl) thio) pyrazin-2-yl) -1, 3-dihydrospiro [ indene-2, 4' -piperidin ] -1-yl) -2-methylpropane-2-sulfinamide
Figure BDA0003005435270000762
6-chloro-3- ((3-chloro-2-morpholinopyridin-4-yl) thio) pyrazin-2-amine (60mg, 0.22mmol), (R) -N- ((S) -1, 3-dihydrospiro [ indene-2, 4' -piperidin ] -1-yl) -2-methylpropane-2-sulfinamide (76mg, 0.25mmol), potassium carbonate (231mg, 1.67mmol) were dissolved in DMF (5mL), stirred with microwave under nitrogen at 100 ℃ for 2h, added with water, and extracted with ethyl acetate. Dried over anhydrous sodium sulfate and concentrated to dryness to give an oil which was used directly in the next reaction.
MS m/z(ESI):628.2[M+H]+.
The fifth step: preparation of (S) -1'- (6-amino-5- ((3-chloro-2-morpholinopyridin-4-yl) thio) pyrazin-2-yl) -1, 3-dihydrospiro [ indene-2, 4' -piperidin ] -1-amine
Figure BDA0003005435270000763
(R) -N- ((S) -1' - (6-amino-5- ((3-chloro-2-morpholinopyridin-4-yl) thio) pyrazin-2-yl) -1, 3-dihydrospiro [ indene-2, 4' -piperidine ] -1-yl) -2-methylpropane-2-sulfinamide (obtained in the previous step) is dissolved in methanol (5mL), 4M HCl/1, 4-dioxane (5mL) is added, stirring is carried out for 1h at room temperature, concentration is carried out till dryness, and column chromatography purification is carried out to obtain the target product (S) -1' - (6-amino-5- ((3-chloro-2-morpholinopyridin-4-yl) thio) pyrazin-2-yl) -1, 3-dihydrospiro [ indene-2, 4' -piperidine ] -1-amine (6mg, 5% over 2 steps).
MS m/z(ESI):524.1[M+H]+.
Example 12
Preparation of (S) -1'- (6-amino-5- ((3-chloro-2- (3, 3-difluoroazetidin-1-yl) pyridin-4-yl) thio) pyrazin-2-yl) -1, 3-dihydrospiro [ indene-2, 4' -piperidin ] -1-amine
Figure BDA0003005435270000771
Example 12 the procedure was as in example 11.
MS m/z(ESI):530.1[M+H]+,532.1[M+2+H]+.
Example 13
Preparation of (S) -1'- (6-amino-5- ((3-chloro-2- (pyrrolidin-1-yl) pyridin-4-yl) thio) pyrazin-2-yl) -1, 3-dihydrospiro [ indene-2, 4' -piperidine ] -1-amine
Figure BDA0003005435270000772
The procedure for the preparation of example 13 is as in example 11.
MS m/z(ESI):508.1[M+H]+,510.1[M+2+H]+.
Example 14
Preparation of (S) -1'- (6-amino-5- ((2- (azetidin-1-yl) -3-chloropyridin-4-yl) thio) pyrazin-2-yl) -4, 6-dihydrospiro [ cyclopenta [ d ] oxazole-5, 4' -piperidine ] -6-amine
Figure BDA0003005435270000773
Example 14 the procedure was as in example 11.
MS m/z(ESI):508.1[M+H]+,510.1[M+2+H]+.
Example 15
Preparation of (S) -1'- (6-amino-5- ((2- (azetidin-1-yl) -3-chloropyridin-4-yl) thio) pyrazin-2-yl) -1-methyl-4, 6-dihydro-1H-spiro [ cyclopenta [ d ] imidazole-5, 4' -piperidine ] -6-amine
Figure BDA0003005435270000781
Example 15 the procedure was as in example 11.
MS m/z(ESI):498.1[M+H]+,500.1[M+2+H]+.
Example 16
Preparation of (S) -1'- (6-amino-5- ((2- (azetidin-1-yl) -3-chloropyridin-4-yl) thio) pyrazin-2-yl) -2-methyl-2, 6-dihydro-4H-spiro [ cyclopenta [ c ] pyrazole-5, 4' -piperidin ] -4-amine
Figure BDA0003005435270000782
Example 16 the procedure was as in example 11.
MS m/z(ESI):498.1[M+H]+,500.1[M+2+H]+.
Example 17
Preparation of (S) -1'- (6-amino-5- ((3-chloro-2-morpholinopyridin-4-yl) thio) -3-methylpyrazin-2-yl) -1, 3-dihydrospiro [ indene-2, 4' -piperidine ] -1-amine
Figure BDA0003005435270000783
The procedure for the preparation of example 17 is as in example 6.
MS m/z(ESI):538.1[M+H]+,540.1[M+2+H]+.
Example 18
Preparation of (S) -1'- (6-amino-5- ((3-chloro-2- (pyrrolidin-1-yl) pyridin-4-yl) thio) -3-methylpyrazin-2-yl) -1, 3-dihydrospiro [ indene-2, 4' -piperidine ] -1-amine
Figure BDA0003005435270000784
Example 18 the procedure was as in example 6.
MS m/z(ESI):522.1[M+H]+,524.1[M+2+H]+.
Example 19
Preparation of (S) -1'- (6-amino-5- ((3-chloro-2- (pyrrolidin-1-yl) pyridin-4-yl) thio) -3-methylpyrazin-2-yl) -4, 6-dihydrospiro [ cyclopenta [ d ] thiazole-5, 4' -piperidine ] -6-amine
Figure BDA0003005435270000791
The procedure for the preparation of example 19 is as described in the experimental protocol of example 6.
MS m/z(ESI):529.1[M+H]+,531.1[M+2+H]+.
Example 20
Preparation of (S) -1'- (6-amino-5- ((3-chloro-2- (pyrrolidin-1-yl) pyridin-4-yl) thio) -3-methylpyrazin-2-yl) -4, 6-dihydrospiro [ cyclopenta [ d ] oxazole-5, 4' -piperidine ] -6-amine
Figure BDA0003005435270000792
The procedure for the preparation of example 20 is as described in the experimental protocol of example 6.
MS m/z(ESI):513.1[M+H]+,515.1[M+2+H]+.
Example 21
Preparation of (S) -1'- (6-amino-5- ((3-chloro-2- (pyrrolidin-1-yl) pyridin-4-yl) thio) -3-methylpyrazin-2-yl) -1-methyl-4, 6-dihydro-1H-spiro [ cyclopenta [ d ] imidazole-5, 4' -piperidine ] -6-amine
Figure BDA0003005435270000793
The procedure for the preparation of example 21 is as described in the experimental protocol of example 6.
MS m/z(ESI):526.1[M+H]+,528.1[M+2+H]+.
Example 22
Preparation of (S) -1'- (6-amino-5- ((3-chloro-2- (pyrrolidin-1-yl) pyridin-4-yl) thio) -3-methylpyrazin-2-yl) -2-methyl-2, 6-dihydro-4H-spiro [ cyclopenta [ c ] pyrazole-5, 4' -piperidin ] -4-amine
Figure BDA0003005435270000801
Example 22 the procedure was as in example 6.
MS m/z(ESI):526.1[M+H]+,528.1[M+2+H]+.
Example 23
Preparation of (S) -1'- (6-amino-5- ((3-chloro-2-morpholinopyridin-4-yl) thio) -3-methylpyrazin-2-yl) -4, 6-dihydrospiro [ cyclopenta [ d ] oxazole-5, 4' -piperidine ] -6-amine
Figure BDA0003005435270000802
The procedure for the preparation of example 23 is as described in the experimental protocol of example 6.
MS m/z(ESI):529.1[M+H]+,531.1[M+2+H]+.
Example 24
Preparation of (S) -1'- (6-amino-5- ((3-chloro-2-morpholinopyridin-4-yl) thio) -3-methylpyrazin-2-yl) -2-methyl-2, 6-dihydro-4H-spiro [ cyclopenta [ c ] pyrazole-5, 4' -piperidin ] -4-amine
Figure BDA0003005435270000803
Example 24 the procedure was as in example 6.
MS m/z(ESI):542.1[M+H]+,544.1[M+2+H]+.
Example 25
Preparation of (S) -1'- (6-amino-5- ((3-chloro-2-cyclopropylpyridin-4-yl) thio) -3-methylpyrazin-2-yl) -4, 6-dihydrospiro [ cyclopenta [ d ] oxazole-5, 4' -piperidine ] -6-amine
Figure BDA0003005435270000804
The procedure for the preparation of example 25 is as in example 6.
MS m/z(ESI):484.1[M+H]+,486.1[M+2+H]+.
Example 26
Preparation of (S) -1'- (6-amino-5- ((3-chloro-2-cyclopropylpyridin-4-yl) thio) pyrazin-2-yl) -4, 6-dihydrospiro [ cyclopenta [ d ] oxazole-5, 4' -piperidine ] -6-amine
Figure BDA0003005435270000811
Example 26 the procedure was as in example 1.
MS m/z(ESI):470.1[M+H]+,472.1[M+2+H]+.
Example 27
Preparation of (S) -1'- (6-amino-5- ((2-cyclopropylpyridin-4-yl) thio) pyrazin-2-yl) -4, 6-dihydrospiro [ cyclopenta [ d ] oxazole-5, 4' -piperidin ] -6-amine
Figure BDA0003005435270000812
Example 27 the procedure was as in example 1.
MS m/z(ESI):436.1[M+H]+.
Example 28
Preparation of (S) -1'- (6-amino-5- ((2-cyclopropyl-3-methylpyridin-4-yl) thio) pyrazin-2-yl) -4, 6-dihydrospiro [ cyclopenta [ d ] oxazole-5, 4' -piperidine ] -6-amine
Figure BDA0003005435270000813
Example 28 the procedure was as in example 1.
MS m/z(ESI):450.1[M+H]+.
Example 29
Preparation of (S) -1'- (6-amino-5- ((2-cyclopropyl-3-fluoropyridin-4-yl) thio) pyrazin-2-yl) -4, 6-dihydrospiro [ cyclopenta [ d ] oxazole-5, 4' -piperidine ] -6-amine
Figure BDA0003005435270000821
Example 29 the procedure was as in example 1.
MS m/z(ESI):454.1[M+H]+.
Example 30
Preparation of (S) -1- (4- ((3-amino-5- (1-amino-1, 3-dihydrospiro [ indene-2, 4 '-piperidin ] -1' -yl) pyrazin-2-yl) thio) -3-chloropyridin-2-yl) azetidin-2-one
Figure BDA0003005435270000822
Example 30 the procedure was as in example 1.
MS m/z(ESI):508.1[M+H]+,510.1[M+2+H]+.
Example 31
Preparation of (S) -1- (4- ((3-amino-5- (1-amino-1, 3-dihydrospiro [ indene-2, 4 '-piperidine ] -1' -yl) pyrazin-2-yl) thio) -3-chloropyridin-2-yl) pyrrolidin-2-one
Figure BDA0003005435270000823
Example 31 the procedure was as in example 1.
MS m/z(ESI):522.1[M+H]+,524.1[M+2+H]+.
Example 32
Preparation of (S) -1- (3- (1-amino-1, 3-dihydrospiro [ indene-2, 4 '-piperidin ] -1' -yl) -6- ((2-cyclopropylpyridin-4-yl) thio) -5-methylpyrazin-2-yl) cyclopropane-1-ol
Figure BDA0003005435270000831
The first step is as follows: preparation of (S) -1- (3- (1-amino-1, 3-dihydrospiro [ indene-2, 4 '-piperidin ] -1' -yl) -6- ((2-cyclopropylpyridin-4-yl) thio) -5-methylpyrazin-2-yl) cyclopropane-1-ol
Figure BDA0003005435270000832
To (S) -3- (1-amino-1, 3-dihydrospiro [ indene-2, 4' -piperidine) at 0 deg.C]Ethyl magnesium bromide (0.1mL,0.30mmol,3M) and tetraisopropyl titanate (43mg,0.15mmol) were added dropwise to a solution of ethyl (1' -yl) -6- (2-cyclopropylpyridin-4-yl) -5-methylpyrazine-2-carboxylate (50mg,0.10mmol) in THF (10mL), and the mixture was warmed to room temperature and stirred for 5 hours. With saturated NH 4The reaction was quenched with aqueous Cl, extracted with dichloromethane (3 x 20mL), dried over anhydrous sodium sulfate, the organic phase concentrated and purified by preparative HPLC to give (S) -1- (3- (1-amino-1, 3-dihydrospiro [ indene-2, 4' -piperidine)]-1' -yl) -6- ((2-cyclopropylpyridin-4-yl) -5-methylpyrazin-2-yl) cyclopropane-1-ol (5mg, yield 10%) as a white solid.
MS m/z(ESI):500.1[M+H]+.
Example 33
Preparation of (S) -1- (3- (1-amino-1, 3-dihydrospiro [ indene-2, 4 '-piperidine ] -1' -yl) -6- ((3-chloro-2-cyclopropylpyridin-4-yl) thio) -5-methylpyrazin-2-yl) cyclopropane-1-ol
Figure BDA0003005435270000833
The procedure for the preparation of example 33 is as described in example 32.
MS m/z(ESI):534.1[M+H]+,536.1[M+2+H]+.
Example 34
Preparation of (S) -1- (3- (1-amino-1, 3-dihydrospiro [ indene-2, 4 '-piperidine ] -1' -yl) -6- ((2-amino-3-chloropyridin-4-yl) thio) -5-methylpyrazin-2-yl) cyclopropane-1-ol
Figure BDA0003005435270000841
Example 34 the procedure was as in example 32.
MS m/z(ESI):509.1[M+H]+,511.1[M+2+H]+.
Example 35
Preparation of (S) -8- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) pyrazin-2-yl) -2-cyclopropyl-8-azaspiro [4.5] dec-2-en-1-amine
Figure BDA0003005435270000842
The first step is as follows: preparation of tert-butyl 4-allyl-4-formylpiperidine-1-carboxylic ester
Figure BDA0003005435270000843
To tert-butyl 4-formylpiperidine-1-carboxylate (30g,14.1mmol) in N, N-dimethylformamide (300mL) was added lithium tert-butoxide (13.5g,16.9mmol) at 0 ℃ and stirred for 30 minutes, followed by addition of allyl bromide (19g,16.2mmol) and stirring at 0 ℃ for 2 hours. The reaction solution was poured into an aqueous ammonium chloride solution (1L), extracted with ethyl acetate (1L. times.2), and the ethyl acetate layer was washed with a saturated aqueous sodium chloride solution (500mL), dried over anhydrous sodium sulfate and purified by column chromatography [ eluent: petroleum ether-ethyl acetate/petroleum ether from 0% to 2% ] to give tert-butyl 4-allyl-4-formylpiperidine-1-carboxylate (15g, yield 42%) as a colorless oily product.
The second step is that: preparation of tert-butyl 4-allyl-4- (1-hydroxyallyl) piperidine-1-carboxylic acid ester
Figure BDA0003005435270000844
To a solution of tert-butyl 4-allyl-4-formylpiperidine-1-carboxylate (25g,99mmol) in tetrahydrofuran (300mL) was added dropwise vinylmagnesium chloride (65mL,123.5mmol) at-78 deg.C, and the mixture was stirred slowly at room temperature for 30 minutes. The reaction solution was poured into an aqueous ammonium chloride solution (1L), extracted with ethyl acetate (1L. times.2), the ethyl acetate layer was washed with a saturated aqueous sodium chloride solution (500mL), dried over anhydrous sodium sulfate and purified by column chromatography [ eluent: petroleum ether-ethyl acetate/petroleum ether from 0% to 5% ] to give tert-butyl 4-allyl-4- (1-hydroxyallyl) piperidine-1-carboxylate (25g, yield 90%) as a colorless oily product.
The third step: preparation of tert-butyl 4-acryloyl-4-allylpiperidine-1-carboxylic acid ester
Figure BDA0003005435270000851
To a solution of tert-butyl 4-allyl-4- (1-hydroxyallyl) piperidine-1-carboxylate (25g,89mmol) in dichloromethane (400mL) at 0 deg.C was added a Starm's reagent (41.5g,98mmol), and the mixture was stirred at 40 deg.C for 1 hour. The reaction mixture was slowly poured into an aqueous sodium hydrogencarbonate/sodium sulfite solution (1/1,1L), extracted with dichloromethane (1L. times.2), the dichloromethane layer was washed with a saturated aqueous sodium chloride solution (500mL), dried over anhydrous sodium sulfate and then spin-dried, followed by addition of n-heptane (200mL), stirring for 5 minutes, filtration of insoluble matter, and concentration of the filtrate gave t-butyl 4-acryloyl-4-allylpiperidine-1-carboxylate (24.8g, yield 100%) as a colorless oily product, which was quickly fed to the next step.
The fourth step: preparation of tert-butyl 1-carbonyl-8-azaspiro [4.5] dec-2-ene-8-carboxylic acid esters
Figure BDA0003005435270000852
Tert-butyl 4-acryloyl-4-allylpiperidine-1-carboxylate (24.8g,89mmol) and dichloro (o-isopropoxyphenylmethylene) (tricyclohexylphosphine) ruthenium (II) (1.2g,1.4mmol) were stirred in toluene (700mL) at 90 ℃ for 2 h. After the reaction solution is cooled, the reaction solution is concentrated and purified by column chromatography (eluent: petroleum ether-ethyl acetate/petroleum ether from 0% to 20%) to obtain a red-black solid product, i.e., tert-butyl 1-carbonyl-8-azaspiro [4.5] dec-2-ene-8-carboxylic ester (13g, 58% yield).
MS m/z(ESI):252.1[M+H]+.
The fifth step: preparation of tert-butyl 2-bromo-1-carbonyl-8-azaspiro [4.5] dec-2-ene-8-carboxylic acid esters
Figure BDA0003005435270000853
To a solution of tert-butyl 1-carbonyl-8-azaspiro [4.5] dec-2-ene-8-carboxylate (13g, 51.851.8 mmol) in acetonitrile (300mL) was added pyridine nitroxide (11g,116mmol), N-bromosuccinimide (28g,157mmol) was added, and the mixture was stirred at 80 ℃ for 18 hours. After the reaction solution is cooled, the reaction solution is concentrated and purified by column chromatography [ eluent: petroleum ether-ethyl acetate/petroleum ether from 0% to 20% ] to obtain a yellow solid product, i.e., tert-butyl 2-bromo-1-carbonyl-8-azaspiro [4.5] dec-2-ene-8-carboxylate (4.2g, yield 25%).
MS m/z(ESI):330.1[M+H]+，332.1[M+2+H]+.
And a sixth step: preparation of tert-butyl 2-cyclopropyl-1-carbonyl-8-azaspiro [4.5] dec-2-ene-8-carboxylic acid esters
Figure BDA0003005435270000854
Tert-butyl 2-bromo-1-carbonyl-8-azaspiro [4.5] dec-2-ene-8-carboxylate (4.2g,12.8mmol), potassium cyclopropyltrifluoroborate (3.79g,25.6mmol), cesium carbonate (12.5g,38.4mmol) and [1,1' -bis (diphenylphosphino) ferrocene ] dichloropalladium dichloromethane complex (731mg,0.9mmol) were stirred in dioxane (100mL) and water (10mL) at 100 ℃ for 18 h. The reaction mixture was quenched with water (150mL), extracted with ethyl acetate (150 mL. times.2), the ethyl acetate layer was washed with saturated aqueous sodium chloride (50mL), dried over anhydrous sodium sulfate and purified by column chromatography [ eluent: petroleum ether-ethyl acetate/petroleum ether from 0% to 12% ] to give tert-butyl 2-cyclopropyl-1-carbonyl-8-azaspiro [4.5] dec-2-ene-8-carboxylate (1g, yield 27%) as a yellow oily product.
MS m/z(ESI):292.2[M+H]+
The seventh step: preparation of tert-butyl (R, Z) -1- ((tert-butylsulfinyl < sulfinyl >) imino) -2-cyclopropyl-8-azaspiro [4.5] dec-2-ene-8-carboxylate
Figure BDA0003005435270000861
Tert-butyl 2-cyclopropyl-1-carbonyl-8-azaspiro [4.5] dec-2-ene-8-carboxylate (1g,3.4mmol) and (R) - (+) -tert-butylsulfinamide (2g,17.2mmol) were stirred in tetraethyl titanate (30mL) at 100 ℃ for 18 hours. After cooling, ethyl acetate (100mL) was added to the reaction mixture, which was poured into water (150mL) to precipitate a large amount of white solid, which was filtered through celite, separated, the organic phase was spin-dried, and purified by column chromatography [ eluent: petroleum ether-ethyl acetate/petroleum ether from 0% to 50% ] to give tert-butyl (R, Z) -1- ((tert-butylsulfinyl < sulfinyl >) imino) -2-cyclopropyl-8-azaspiro [4.5] dec-2-ene-8-carboxylate (190mg, yield 14%) as a yellow oily product.
MS m/z(ESI):395.2[M+H]+.
Eighth step: preparation of tert-butyl (S) -1- (((R) -tert-butylsulfinyl < sulfinyl >) amino) -2-cyclopropyl-8-azaspiro [4.5] dec-2-ene-8-carboxylic acid ester
Figure BDA0003005435270000862
To tert-butyl (R, Z) -1- ((tert-butylsulfinyl < sulfinyl >) imino) -2-cyclopropyl-8-azaspiro [4.5] dec-2-ene-8-carboxylate (210mg,0.53mmol) in tetrahydrofuran (10mL) was added dropwise diisobutylaluminum hydride (0.9mL,1mmol) at-78 ℃ and stirring was completed for 15 minutes. The reaction was quenched with sodium sulfate decahydrate at-78 ℃ for 10 minutes, stirred, filtered, and the filtrate was concentrated and purified by column chromatography [ eluent: petroleum ether-ethyl acetate/petroleum ether from 0% to 5% ] to give tert-butyl (S) -1- (((R) -tert-butylsulfinyl < sulfinyl >) amino) -2-cyclopropyl-8-azaspiro [4.5] dec-2-ene-8-carboxylate (105mg, 50% yield) as a colorless oily product.
MS m/z(ESI):397.2[M+H]+.
The ninth step: preparation of (S) -2-cyclopropyl-8-azaspiro [4.5] dec-2-en-1-amine
Figure BDA0003005435270000863
To a solution of tert-butyl (S) -1- (((R) -tert-butylsulfinyl < sulfinyl >) amino) -2-cyclopropyl-8-azaspiro [4.5] dec-2-ene-8-carboxylate (105mg,0.27mmol) in methanol (3mL) was added dioxane hydrochloride (15mL,60mmol), and the mixture was stirred at room temperature for 1 hour. The reaction solution was spin-dried to give (S) -2-cyclopropyl-8-azaspiro [4.5] dec-2-en-1-amine (105mg, yield 100%) as a white solid.
MS m/z(ESI):193.2[M+H]+.
The tenth step: preparation of (S) -8- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) pyrazin-2-yl) -2-cyclopropyl-8-azaspiro [4.5] dec-2-en-1-amine
Figure BDA0003005435270000871
3- ((2-amino-3-chloropyridin-4-yl) thio) -6-chloropyrazin-2-amine (76mg,0.26mmol), (S) -2-cyclopropyl-8-azaspiro [4.5] dec-2-en-1-amine (80mg,0.2mmol) and potassium carbonate (167mg,1.2mmol) were stirred in N, N-dimethylformamide (8mL) at 100 ℃ for 7 hours. To the reaction solution was added water (60mL), extracted with ethyl acetate (50mL × 2), and the ethyl acetate layer was washed with saturated aqueous sodium chloride (20mL), dried over anhydrous sodium sulfate and purified over a plate [ eluent: dichloromethane/methanol ═ 7/1] to give (S) -8- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) pyrazin-2-yl) -2-cyclopropyl-8-azaspiro [4.5] dec-2-en-1-amine (37mg, yield 41%) as a yellow oily product.
1H NMR(400MHz,MeOD)δ7.67–7.51(m,2H),5.92(d,J＝8Hz,1H),5.27(s,1H),4.26–4.04(m,2H),3.33(s,1H),3.27–3.08(m,2H),2.44–2.18(m,2H),1.81–1.69(m,1H),1.68–1.47(m,2H),1.42–1.24(m,2H),0.81–0.60(m,2H),0.57–0.28(m,2H).
MS m/z(ESI):444.1[M+H]+，446.1[M+2+H]+.
Example 36
Preparation of (S) -8- (5- ((2-amino-3-chloropyridin-4-yl) thio) pyrazin-2-yl) -2-cyclopropyl-8-azaspiro [4.5] dec-2-en-1-amine
Figure BDA0003005435270000872
The first step is as follows: preparation of 3-chloro-4- ((5-chloropyrazin-2-yl) thio) pyridin-2-amine
Figure BDA0003005435270000873
Potassium 2-amino-3-chloropyridine-4-thiolate (1g, 2.64mmol), 2-bromo-5-chloropyrazine (597mg, 3.11mmol), tris (dibenzylideneacetone) dipalladium (284mg, 0.311mmol), chloro [ (4, 5-bis (diphenylphosphino) -9, 9-dimethylxanthene) -2- (2-aminobiphenyl) ] palladium (II) (360mg, 0.622mmol) and N, N-diisopropylethylamine (1.2g, 9.33mmol) were stirred in N, N-dimethylformamide (10mL) at 90 ℃ for 1 hour, and after spin-drying column chromatography [ eluent: petroleum ether-ethyl acetate/petroleum ether from 0% to 30% ] was purified to give the white solid product 3-chloro-4- ((5-chloropyrazin-2-yl) thio) pyridin-2-amine (250mg, yield 35%).
MS m/z(ESI):273.0[M+H]+，275.0[M+2+H]+。
The second step is that: preparation of (S) -8- (5- ((2-amino-3-chloropyridin-4-yl) thio) pyrazin-2-yl) -2-cyclopropyl-8-azaspiro [4.5] dec-2-en-1-amine
Figure BDA0003005435270000881
3-chloro-4- ((5-chloropyrazin-2-yl) thio) pyridin-2-amine (100mg,0.368mmol), (S) -2-cyclopropyl-8-azaspiro [4.5] dec-2-en-1-amine hydrochloride (105mg,0.265mmol) and potassium carbonate (254mg,1.84mmol) were stirred in N, N-dimethylformamide (8mL) at 100 ℃ for 6 hours. Water (40mL) was added to the reaction solution, extracted with ethyl acetate (40mL × 2), the organic phase was concentrated and purified by thin layer chromatography (dichloromethane/methanol ═ 10/1) to give (S) -8- (5- ((2-amino-3-chloropyridin-4-yl) thio) pyrazin-2-yl) -2-cyclopropyl-8-azaspiro [4.5] dec-2-en-1-amine (40mg, 35% yield) as a yellow solid product.
1H NMR(400MHz,MeOD)δ8.29(d,J＝16Hz,2H),7.60(d,J＝4Hz,1H),5.93(d,J＝4Hz,1H),5.31(s,1H),4.34–4.14(m,2H),3.43–3.35(m,2H),3.28–3.21(m,1H),2.45–2.21(m,2H),1.86–1.49(m,3H),1.44–1.22(m,2H),0.82–0.61(m,2H),0.58–0.30(m,2H).
MS m/z(ESI):429.1[M+H]+，431.1[M+2+H]+.
Example 37
Preparation of (S) -8- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) -3-methylpyrazin-2-yl) -2-methyl-8-azaspiro [4.5] dec-2-en-1-amine
Figure BDA0003005435270000882
Example 37 the procedure was as in example 6.
MS m/z(ESI):432.1[M+H]+,434.1[M+2+H]+.
Example 38
Preparation of (S) -8- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) -3-methylpyrazin-2-yl) -2-ethyl-8-azaspiro [4.5] dec-2-en-1-amine
Figure BDA0003005435270000883
Example 38 the procedure was as in example 6.
MS m/z(ESI):446.1[M+H]+,448.1[M+2+H]+.
Example 39
Preparation of (S) -8- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) -3-methylpyrazin-2-yl) -2-isopropyl-8-azaspiro [4.5] dec-2-en-1-amine
Figure BDA0003005435270000891
The procedure for the preparation of example 39 is as described in the experimental protocol of example 6.
MS m/z(ESI):460.1[M+H]+,462.1[M+2+H]+.
Example 40
Preparation of (S) -8- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) -3-methylpyrazin-2-yl) -2-cyclopropyl-8-azaspiro [4.5] dec-2-en-1-amine
Figure BDA0003005435270000892
The procedure for the preparation of example 40 is as described in the experimental protocol of example 6.
MS m/z(ESI):458.1[M+H]+,460.1[M+2+H]+.
EXAMPLE 41
Preparation of (S) -1- (6- ((2-amino-3-chloropyridin-4-yl) thio) -3- (6-amino-4, 6-dihydrospiro [ cyclopenta [ d ] oxazole-5, 4 '-piperidin ] -1' -yl) -5-methylpyrazin-2-yl) cyclopropane-1-ol
Figure BDA0003005435270000893
Example 41 reference is made to the experimental protocol of example 32.
MS m/z(ESI):500.1[M+H]+,502.1[M+2+H]+.
Example 42
Preparation of (R) -1- (6- ((2-amino-3-chloropyridin-4-yl) thio) -3- (1-amino-8-azaspiro [4.5] dec-2-en-8-yl) -5-methylpyrazin-2-yl) cyclopropane-1-ol
Figure BDA0003005435270000901
Example 42 reference example 32 protocol was used for the preparation of the reference.
MS m/z(ESI):459.1[M+H]+,461.1[M+2+H]+.
Example 43
Preparation of (S) -1- (3- (1-amino-2-methyl-8-azaspiro [4.5] dec-2-en-8-yl) -6- ((2-amino-3-chloropyridin-4-yl) thio) -5-methylpyrazin-2-yl) cyclopropane-1-ol
Figure BDA0003005435270000902
Example 43 the procedure was as in example 32.
MS m/z(ESI):473.1[M+H]+,475.1[M+2+H]+.
Example 44
Preparation of (S) -1- (3- (1-amino-2-cyclopropyl-8-azaspiro [4.5] dec-2-en-8-yl) -6- ((2-amino-3-chloropyridin-4-yl) thio) -5-methylpyrazin-2-yl) cyclopropane-1-ol
Figure BDA0003005435270000903
Example 44 the procedure was as in example 32.
MS m/z(ESI):499.1[M+H]+,501.1[M+2+H]+.
Example 45
Preparation of (S) - (3- (1-amino-1, 3-dihydrospiro [ indene-2, 4 '-piperidine ] -1' -yl) -6- ((2-amino-3-chloropyridin-4-yl) thio) -5-methylpyrazin-2-yl) methanol
Figure BDA0003005435270000904
The first step is as follows: preparation of 2-ethylhexyl 3- ((2-amino-3-chloropyridin-4-yl) thio) propionate
Figure BDA0003005435270000911
3-chloro-4-iodopyridin-2-amine (3g, 11.8mmol), 2-ethylhexyl 3-mercaptopropionate (2.9g, 14.2mmol), palladium acetate (132mg, 0.59mmol), 4, 5-bis (diphenylphosphino) -9, 9-dimethylxanthene (683mg,1.18mmol) and N, N-diisopropylethylamine (561mg,4.3mmol) were stirred in dioxane (80mL) at 100 ℃ for 18 hours. The reaction solution was filtered, the filtrate was concentrated and purified by column chromatography [ eluent: petroleum ether-petroleum ether/ethyl acetate 60:40) ] to give 2-ethylhexyl 3- ((2-amino-3-chloropyridin-4-yl) thio) propionate as a pale yellow solid (3.8g, 94% yield).
MS m/z(ESI):345.1[M+H]+，347.0[M+2+H]+.
The second step is that: preparation of potassium 2-amino-3-chloropyridine-4-mercaptide
Figure BDA0003005435270000912
Add potassium tert-butoxide (174mg,1.53mmol) to a solution of 2-ethylhexyl 3- ((2-amino-3-chloropyridin-4-yl) thio) propionate (500mg,1.5mmol) in EtOH (30mL) and stir at 40 ℃ for 2 h. The reaction solution was concentrated to give 2-amino-3-chloropyridine-4-potassium mercaptide (500mg) as a yellow solid, and the crude product was directly fed to the next step.
MS m/z(ESI):161.0[M+H]+，163.0[M+2+H]+.
The third step: preparation of ethyl 6-bromo-3- ((S) -1- (((R) -tert-butylsulfinyl < sulfinyl >) amino) -1, 3-dihydrospiro [ indene-2, 4 '-piperidin ] -1' -yl) -5-methylpyrazine-2-carboxylate
Figure BDA0003005435270000913
6-bromo-3-chloro-5-methylpyrazine-2-carboxylic acid ethyl ester (840mg,3mmol), (S) -N- (1, 3-dihydrospiro [ indene-2, 4' -piperidin ] -1-yl) pivaloyl-indene dione amide trifluoroacetate (1.5g,3.6mmol) and potassium carbonate (1.7g,12mmol) were stirred in acetonitrile (40mL) at 55 ℃ for 18 h. The reaction solution was filtered, the filtrate was concentrated and purified by column chromatography [ eluent: petroleum ether-petroleum ether/ethyl acetate 60:40) ] to give 6-bromo-3- ((S) -1- (((R) -tert-butylsulfinyl < sulfinyl >) amino) -1, 3-dihydrospiro [ indene-2, 4 '-piperidin ] -1' -yl) -5-methylpyrazine-2-carboxylic acid ethyl ester as a yellow solid (1.5g, 91% yield).
MS m/z(ESI):549.2[M+H]+，551.2[M+2+H]+.
The fourth step: 6- ((2-amino-3-chloropyridin-4-yl) thio) -3- ((S) -1- (((R) -tert-butylsulfinyl)<Sulfinyl radical>) Amino) -1, 3-dihydrospiro [ indene-2, 4' -piperidine]Preparation of (E) -1' -yl) -5-methylpyrazine-2-carboxylic acid ethyl ester
Figure BDA0003005435270000921
6-bromo-3- ((S) -1- (((R) -tert-butylsulfinyl < sulfinyl >) amino) -1, 3-dihydrospiro [ indene-2, 4 '-piperidine ] -1' -yl) -5-methylpyrazine-2-carboxylic acid ethyl ester (798mg,1.45mmol), potassium 2-amino-3-chloropyridine-4-mercaptide (500mg,1.45mmol, crude), tris (dibenzylideneacetone) dipalladium (66mg,0.073mmol), 4, 5-bis (diphenylphosphino) -9, 9-dimethylxanthene (84mg,0.145mmol) and N, N-diisopropylethylamine (561mg,4.35mmol) in N, N-dimethylformamide (15mL) were stirred with microwave at 100 ℃ for 1.5 hours. The reaction solution was filtered, and the filtrate was concentrated and purified by column chromatography [ eluent: dichloromethane to dichloromethane/methanol (98.5:1.5) ] to give ethyl 6- ((2-amino-3-chloropyridin-4-yl) thio) -3- ((S) -1- (((R) -tert-butylsulfinyl < sulfinyl >) amino) -1, 3-dihydrospiro [ indene-2, 4 '-piperidin ] -1' -yl) -5-methylpyrazine-2-carboxylate as a yellow solid (500mg, 55% yield).
MS m/z(ESI):629.1[M+H]+，631.1[M+2+H]+.
The fifth step: preparation of (R) -N- ((S) -1'- (5- ((2-amino-3-chloropyridin-4-yl) thio) -3- (hydroxymethyl) -6-methylpyrazin-2-yl) -1, 3-dihydrospiro [ indene-2, 4' -piperidin ] -1-yl) -2-methylpropane-2-sulfinamide
Figure BDA0003005435270000922
Lithium aluminum tetrahydrochloride (0.7mL, 2.5M) was added dropwise to a solution of ethyl 6- ((2-amino-3-chloropyridin-4-yl) thio) -3- ((S) -1- (((R) -tert-butylsulfinyl < sulfinyl >) amino) -1, 3-dihydrospiro [ indene-2, 4 '-piperidin ] -1' -yl) -5-methylpyrazine-2-carboxylate (500mg,0.795mmol) in tetrahydrofuran (30mL) at 0 ℃ and stirred at 0 ℃ for 30 minutes. The reaction was quenched with sodium sulfate decahydrate and stirred for 10 minutes. Filtration through celite, washing of the filter cake twice with tetrahydrofuran (50mL), concentration of the filtrate, and purification by column chromatography [ eluent: dichloromethane to dichloromethane/methanol (95:5) ] gave (R) -N- ((S) -1'- (5- ((2-amino-3-chloropyridin-4-yl) thio) -3- (hydroxymethyl) -6-methylpyrazin-2-yl) -1, 3-dihydrospiro [ indene-2, 4' -piperidin ] -1-yl) -2-methylpropane-2-sulfinamide as a yellow solid (220mg, 47% yield).
MS m/z(ESI):587.1[M+H]+，589.1[M+2+H]+.
And a sixth step: preparation of (S) - (3- (1-amino-1, 3-dihydrospiro [ indene-2, 4 '-piperidine ] -1' -yl) -6- ((2-amino-3-chloropyridin-4-yl) thio) -5-methylpyrazin-2-yl) methanol
Figure BDA0003005435270000931
Add dioxane hydrochloride (20mL, 4.0M) to a solution of (R) -N- ((S) -1'- (5- ((2-amino-3-chloropyridin-4-yl) thio) -3- (hydroxymethyl) -6-methylpyrazin-2-yl) -1, 3-dihydrospiro [ indene-2, 4' -piperidin ] -1-yl) -2-methylpropane-2-sulfinamide (220mg,0.375mmol) in methanol (3mL) and stir at room temperature for 30 min. The reaction solution was concentrated, and then dissolved in methanol (10mL), adjusted to alkali with ammonia water, and purified by column chromatography [ eluent: dichloromethane to dichloromethane/methanol (1% NH3-H2O) (95:5) ] after spin-drying to obtain (S) - (3- (1-amino-1, 3-dihydrospiro [ indene-2, 4 '-piperidin ] -1' -yl) -6- ((2-amino-3-chloropyridin-4-yl) thio) -5-methylpyrazin-2-yl) methanol (115mg, yield 63%) as a white solid.
1H NMR(400MHz,MeOD)δ7.59(d,J＝4Hz,1H),7.41–7.34(m,1H),7.27–7.16(m,3H),5.89(d,J＝4Hz,1H),4.64(s,2H),4.00(s,1H),3.88(d,J＝12Hz,2H),3.28–3.20(m,2H),3.14(d,J＝16Hz,1H),2.82(d,J＝16Hz,1H),2.48(s,3H),2.01–1.85(m,2H),1.67–1.58(m,1H),1.52–1.43(m,1H).
MS m/z(ESI):483.2[M+H]+，485.2[M+2+H]+.
Example 46
Preparation of (S) - (6- ((2-amino-3-chloropyridin-4-yl) thio) -3- (6-amino-4, 6-dihydrospiro [ cyclopenta [ d ] thiazol-5, 4 '-piperidin ] -1' -yl) -5-methylpyrazin-2-yl) methanol
Figure BDA0003005435270000932
The first step is as follows: preparation of ethyl 6-bromo-3- ((S) -6- (((R) -tert-butylsulfinyl < sulfinyl >) amino) -4, 6-dihydrospiro [ cyclopenta [ d ] thiazole-5, 4 '-piperidin ] -1' -yl) -5-methylpyrazine-2-carboxylate
Figure BDA0003005435270000933
(R) -N- ((S) -4, 6-dihydrospiro [ cyclopenta [ d ] thiazole-5, 4' -piperidin ] -6-yl) -2-methylpropane-2-sulfinamide (470mg, 1.5mmol, 1.0eq), ethyl 6-bromo-3-chloro-5-methylpyrazine-2-carboxylate (398mg,1.43mmol,0.95eq) and potassium carbonate (1.04g,7.5mmol,5.0eq) were dissolved in DMF (4.0 mL). Heating to 60 ℃ under the protection of nitrogen, and stirring for reaction for 3 hours. At the end of the reaction, the DMF was removed by concentration and the crude product purified by column chromatography (100% EtOAc in Petro ether) to give ethyl 6-bromo-3- ((S) -6- (((R) -tert-butylsulfinyl < sulfinyl >) amino) -4, 6-dihydrospiro [ cyclopenta [ d ] thiazole-5, 4 '-piperidin ] -1' -yl) -5-methylpyrazine-2-carboxylate (600mg, 72% yield) as a yellow solid.
MS m/z(ESI):556.1[M+H]+,558.1[M+H+2]+.
The second step is that: preparation of ethyl 6- ((2-amino-3-chloropyridin-4-yl) thio) -3- ((S) -6- (((R) -tert-butylsulfinyl < sulfinyl >) amino) -4, 6-dihydrospiro [ cyclopenta [ d ] thiazol-5, 4 '-piperidin ] -1' -yl) -5-methylpyrazine-2-carboxylate
Figure BDA0003005435270000941
Ethyl 6-bromo-3- ((S) -6- (((R) -tert-butylsulfinyl)<Sulfinyl radical>) Amino) -4, 6-dihydrospiro [ cyclopenta [ d ]]Thiazole-5, 4' -piperidines]-1' -yl) -5-methylpyrazine-2-carboxylate (600mg, 1.08mmol, 1.0eq), potassium 2-amino-3-chloropyridine-4-thiolate (536mg, 2.70mmol, 2.5eq) dissolved in 10mL DMF, placed in a microwave reaction tube and tris (dibenzylideneacetone) dipalladium (50mg, 0.054mmol, 0.05eq), Xantphos (62mg, 0.11mmol, 0.1eq) and DIPEA (419mg, 3.24mmol, 3.0eq) bubbled with nitrogen for 3 min. Heating the reaction solution to 105 ℃ by microwave to react for 75 min; after the reaction solution was cooled to room temperature, it was diluted with 20mL of ethyl acetate, filtered through celite, concentrated, and purified by column chromatography (10% MeOH in CH)2Cl2) To obtain a yellow solid product, namely ethyl 6- ((2-amino-3-chloropyridin-4-yl) thio) -3- ((S) -6- (((R) -tert-butylsulfinyl)<Sulfinyl radical>) Amino) -4, 6-dihydrospiro [ cyclopenta [ d ]]Thiazole-5, 4' -piperidines]-1' -yl) -5-methylpyrazine-2-carboxylic acid ester (360mg, 52% yield).
MS m/z(ESI):636.0[M+H]+,638.0[M+H+2]+.
The third step: preparation of ethyl 6- ((2-amino-3-chloropyridin-4-yl) thio) -3- ((S) -6- (((R) -tert-butylsulfinyl < sulfinyl >) amino) -4, 6-dihydrospiro [ cyclopenta [ d ] thiazol-5, 4 '-piperidin ] -1' -yl) -5-methylpyrazine-2-carboxylate
Figure BDA0003005435270000942
Ethyl 6- ((2-amino-3-chloropyridin-4-yl) thio) -3- ((S) -6- (((R) -tert-butylsulfinyl)<Sulfinyl radical>) Amino) -4, 6-dihydrospiro [ cyclopenta [ d ]]Thiazole-5, 4' -piperidines]-1' -yl) -5-methylpyrazine-2-carboxylate (360mg, 0.57mmol, 1.0eq) was dissolved in 40mL of absoluteTHF water, nitrogen replacement protection. Cooling to 0 deg.C, and adding LiAlH dropwise4Tetrahydrofuran solution (0.6mL,1.14mmol,2.0eq., D ═ 2M). After the addition, the temperature was slowly raised to room temperature and stirred for 3 hours. After the reaction, the reaction mixture was diluted with ethyl acetate (100mL), cooled to 0 deg.C, 2mL of water was added dropwise, 4mL of 15% aqueous sodium hydroxide solution was added, 6mL of water was added, the mixture was warmed to room temperature and stirred for 30 minutes, an appropriate amount of anhydrous sodium sulfate was added, stirring was carried out for 15 minutes, the solid was removed by filtration, the filtrate was concentrated, and the crude product was purified by column chromatography (10% MeOH in CH)2Cl2) To give ethyl 6- ((2-amino-3-chloropyridin-4-yl) thio) -3- ((S) -6- (((R) -tert-butylsulfinyl) as a yellow solid<Sulfinyl radical>) Amino) -4, 6-dihydrospiro [ cyclopenta [ d ]]Thiazole-5, 4' -piperidines]-1' -yl) -5-methylpyrazine-2-carboxylic acid ester (150mg, yield 44%).
MS m/z(ESI):594.1[M+H]+,596.1[M+H+2]+.
The fourth step: preparation of (S) - (6- ((2-amino-3-chloropyridin-4-yl) thio) -3- (6-amino-4, 6-dihydrospiro [ cyclopenta [ d ] thiazol-5, 4 '-piperidin ] -1' -yl) -5-methylpyrazin-2-yl) methanol
Figure BDA0003005435270000951
Ethyl 6- ((2-amino-3-chloropyridin-4-yl) thio) -3- ((S) -6- (((R) -tert-butylsulfinyl)<Sulfinyl radical>) Amino) -4, 6-dihydrospiro [ cyclopenta [ d ]]Thiazole-5, 4' -piperidines]-1' -yl) -5-methylpyrazine-2-carboxylate (150mg, 0.25mmol, 1.0eq) was dissolved in MeOH (5mL), and a dioxane solution of hydrochloric acid (1.0mL,4M) was added and reacted at room temperature for 1 hour. The reaction mixture was concentrated to dryness, dissolved in 5mL of methanol, and then dissolved in 7M NH3The reaction solution was then adjusted to alkaline pH, the reaction was concentrated and purified using a normal phase column (10% MeOH in CH)2Cl2) Then purified by reverse phase column (30% MeCN in H)2O(0.1％NH3.H2O)) to obtain a gray solid target compound (S) - (6- ((2-amino-3-chloropyridin-4-yl) thio) -3- (6-amino-4, 6-dihydrospiro [ cyclopenta [ d ]]Thiazole-5, 4' -piperidines]-1' -yl) -5-methylpyrazin-2-yl methanol (52mg,yield 42%).
1H NMR(400MHz,DMSO)δ8.93(s,1H),7.62(s,1H),6.31(s,1H),5.72(s,1H),5.41(s,1H),4.45(s,2H),3.98(s,1H),3.84(s,2H),2.76(dd,J＝47.2,16.9Hz,4H),2.37(s,3H),1.60(dd,J＝165.2,116.4Hz,7H).
MS m/z(ESI):490.1[M+H]+,492.1[M+H+2]+.
Example 47
Preparation of (S) - (6- ((2-amino-3-chloropyridin-4-yl) thio) -3- (6-amino-4, 6-dihydrospiro [ cyclopenta [ d ] oxazole-5, 4 '-piperidin ] -1' -yl) -5-methylpyrazin-2-yl) methanol
Figure BDA0003005435270000952
Example 47 the procedure was as in example 46.
MS m/z(ESI):474.1[M+H]+,476.1[M+2+H]+.
Example 48
Preparation of (R) -8- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) pyrazin-2-yl) -2-methylene-8-azaspiro [4.5] decan-1-amine
Figure BDA0003005435270000961
Example 48 the procedure was as described in example 1.
MS m/z(ESI):418.1[M+H]+,420.1[M+2+H]+.
Example 49
Preparation of (S, E) -8- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) pyrazin-2-yl) -2-ethylidene-8-azaspiro [4.5] decan-1-amine
Figure BDA0003005435270000962
Example 49 the procedure is as in example 1.
MS m/z(ESI):432.1[M+H]+,434.1[M+2+H]+.
Example 50
Preparation of (S) -8- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) pyrazin-2-yl) -2- (propan-2-ylidene) -8-azaspiro [4.5] decan-1-amine
Figure BDA0003005435270000963
Example 50 the procedure was as in example 1.
MS m/z(ESI):446.1[M+H]+,448.1[M+2+H]+.
Example 51
Preparation of (S, E) -8- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) -3-methylpyrazin-2-yl) -2-ethylidene-8-azaspiro [4.5] decan-1-amine
Figure BDA0003005435270000964
Example 51 the procedure was followed according to the experimental protocol of example 6.
MS m/z(ESI):446.1[M+H]+,448.1[M+2+H]+.
Example 52
Preparation of (R) -8- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) -3-methylpyrazin-2-yl) -2-methylene-8-azaspiro [4.5] decan-1-amine
Figure BDA0003005435270000971
Example 52 the procedure was as in example 6.
MS m/z(ESI):432.1[M+H]+,434.1[M+2+H]+.
Example 53
Preparation of (S) -1'- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) -3-fluoropyrazin-2-yl) -1, 3-dihydrospiro [ indene-2, 4' -piperidine ] -1-amine
Figure BDA0003005435270000972
The first step is as follows: preparation of (R) -N- ((S) -1'- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) pyrazin-2-yl) -1, 3-dihydrospiro [ indene-2, 4' -piperidin ] -1-yl) -2-methylpropane-2-sulfinamide
Figure BDA0003005435270000973
3- ((2-amino-3-chloropyridin-4-yl) thio) -6-chloropyrazin-2-amine (200mg,0.694mmol), (R) -N- ((S) -1, 3-dihydrospiro [ indene-2, 4' -piperidine]-1-yl) -2-methylpropane-2-sulfinamide (350mg,0.833mmol, [ alpha ]]20 D1.773) and potassium carbonate (479mg,3.47mmol) in N, N-dimethylformamide (7mL) was stirred at 100 ℃ for 8 hours. After the reaction solution was cooled, 40mL of water was added, and extraction was performed with ethyl acetate (40 mL. times.2); the ethyl acetate layer was washed with a saturated aqueous solution of sodium chloride (20mL), dried over anhydrous sodium sulfate and subjected to column chromatography [ eluent: dichloromethane to dichloromethane/methanol from 0% to 3%]Purifying to obtain an orange solid product (R) -N- ((S) -1'- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) pyrazin-2-yl) -1, 3-dihydrospiro [ indene-2, 4' -piperidine]-1-yl) -2-methylpropane-2-sulfinamide (200mg, 52% yield).
MS m/z(ESI):558.2[M+H]+。
The second step is that: preparation of (S) -1'- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) pyrazin-2-yl) -1, 3-dihydrospiro [ indene-2, 4' -piperidine ] -1-amine
Figure BDA0003005435270000981
Add dioxane hydrochloride solution (20mL,80mmol) to a solution of (R) -N- ((S) -1'- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) pyrazin-2-yl) -1, 3-dihydrospiro [ indene-2, 4' -piperidin ] -1-yl) -2-methylpropane-2-sulfinamide (188mg, 0.337mmol) in methanol (5mL) and stir at room temperature for 1 h. The reaction was concentrated to dryness, dissolved in 5mL of methanol, pH adjusted to basic with ammonia, and purified by column chromatography [ eluent: dichloromethane to dichloromethane/methanol from 0% to 7.5% ] to give (S) -1'- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) pyrazin-2-yl) -1, 3-dihydrospiro [ indene-2, 4' -piperidin ] -1-amine as a white solid (130mg, 85% yield).
MS m/z(ESI):454.2[M+H]+。
The third step: preparation of (S) -1'- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) -3-fluoropyrazin-2-yl) -1, 3-dihydrospiro [ indene-2, 4' -piperidine ] -1-amine
Figure BDA0003005435270000982
To a solution of (S) -1'- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) pyrazin-2-yl) -1, 3-dihydrospiro [ indene-2, 4' -piperidin ] -1-amine (100mg,0.22mmol) in acetonitrile (10mL) at-10 deg.C was added N-fluorobisbenzenesulfonamide (69mg,0.22mmol), and the mixture was allowed to slowly warm to room temperature and stirred for 7 hours. The reaction solution was dried by spinning and purified by thin layer chromatography (dichloromethane/methanol ═ 10/1) to give (S) -1'- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) -3-fluoropyrazin-2-yl) -1, 3-dihydrospiro [ indene-2, 4' -piperidine ] -1-amine as an off-white solid (28mg, 27% yield).
1H NMR(400MHz,MeOD)δ7.62(d,J＝4Hz,1H),7.42–7.33(m,1H),7.26–7.15(m,3H),6.00(d,J＝4Hz,1H),4.67–4.54(m,2H),4.30(d,J＝16Hz,2H),3.97(s,1H),3.16(d,J＝16Hz,1H),2.82(d,J＝16Hz,1H),1.95–1.78(m,2H),1.65–1.53(m,1H),1.48–1.38(m,1H).
MS m/z(ESI):472.1[M+H]+，474.1[M+2+H]+.
[α]20 D＝-36.893.
Example 54
(S) -1'- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) -3-fluoropyrazin-2-yl) -5, 7-dihydrospiro [ cyclopenta [ b ] pyridine-6, 4' -piperidine ] -5-amine
Figure BDA0003005435270000991
The first step is as follows: preparation of 3-bromo-2- (chloromethyl) pyridine
Figure BDA0003005435270000992
(3-bromopyridin-2-yl) methanol (7.5g,40mmol) was dissolved in dichloromethane (20mL), and after the reaction was mixed well, SOCl was added dropwise under ice bath conditions2(9.5g,80mmol) and stirred for 1 hour. The reaction mixture was concentrated under reduced pressure, and the obtained crude product was separated and purified by flash silica gel column chromatography to give the objective compound, 3-bromo-2- (chloromethyl) pyridine (5g, yield: 61%), as a colorless liquid.
MS m/z(ESI):205.9[M+H]+.
The second step is that: preparation of 1- (tert-butyl) 4-methyl 4- ((3-bromopyridin-2-yl) methyl) piperidine-1, 4-dicarboxylic acid ester
Figure BDA0003005435270000993
1- (tert-butyl) 4-methylpiperidine-1, 4-dicarboxylate (7.7g, 31.7mmol) and LDA (18.3mL, 36.6mmol) were added to THF (20mL), the reaction mixture was reacted at-60 ℃ for 1 hour, 3-bromo-2- (chloromethyl) pyridine (5g,24.4mmol) was added to the reaction mixture, the reaction was continued at constant temperature for 2 hours, and the mixture was concentrated under reduced pressure, and the resulting crude product was isolated and purified by flash chromatography on a silica gel column to give the objective 1- (tert-butyl) 4-methyl 4- ((3-bromopyridin-2-yl) methyl) piperidine-1, 4-dicarboxylate (9g, yield: 89%) as a pale yellow solid.
MS m/z(ESI):413.0[M+H]+.
The third step: preparation of tert-butyl 5-carbonyl-5, 7-dihydrospiro [ cyclopenta [ b ] pyridine-6, 4 '-piperidine ] -1' -carboxylic acid ester
Figure BDA0003005435270000994
THF (100mL) was added to 1- (tert-butyl) 4-methyl 4- ((3-bromopyridin-2-yl) methyl) piperidine-1, 4-dicarboxylate (9g, 21.84mmol), the reaction mixture was cooled to-78 deg.C, then N-butyllithium solution (1.6N, 17mL) was slowly dropped into the reaction mixture, the reaction was continued at-78 deg.C for 1 hour, followed by addition of aqueous ammonium chloride solution for quenching, extraction with ethyl acetate, drying of the organic phase over anhydrous sodium sulfate, filtration, and concentration under reduced pressure. The crude product was isolated and purified by flash column chromatography on silica gel to give t-butyl 5-carbonyl-5, 7-dihydrospiro [ cyclopenta [ b ] pyridine-6, 4 '-piperidine ] -1' -carboxylate (3g, yield: 45%) as a pale yellow solid.
MS m/z(ESI):303.1[M+H]+.
The fourth step: preparation of tert-butyl (R, Z) -5- ((tert-butylsulfinyl < sulfinyl >) imino) -5, 7-dihydrospiro [ cyclopenta [ b ] pyridine-6, 4 '-piperidine ] -1' -carboxylate
Figure BDA0003005435270001001
Adding tert-butyl 5-carbonyl-5, 7-dihydrospiro [ cyclopenta [ b ] pyridine-6, 4 '-piperidine ] -1' -carboxylic ester (3g,9.93mmol), (R) -2-methylpropane-2-sulfinamide (3.6g,29.8mmol) into ethyl titanate (50mL), stirring the reaction solution uniformly, reacting at 110 ℃ for 13 hours, cooling the reaction temperature to normal temperature, slowly dropwise adding water (100mL) to generate a solid, adding ethyl acetate (200mL), filtering, concentrating the organic phase under reduced pressure to obtain the target compound tert-butyl (R, Z) -5- ((tert-butylsulfinyl < sulfinyl >) imino) -5, 7-dihydrospiro [ cyclopenta [ b ] pyridine-6, 4 '-piperidine ] -1' -carboxylic ester (3g, yield: 75%) was used directly in the next reaction.
MS m/z(ESI):406.1[M+H]+.
The fifth step: preparation of tert-butyl (S) -5- (((R) -tert-butylsulfinyl < sulfinyl >) amino) -5, 7-dihydrospiro [ pentadieno [ b ] pyridine-6, 4 '-piperidine ] -1' -carboxylic acid ester
Figure BDA0003005435270001002
Reacting tert-butyl (R, Z) -5- ((tert-butylsulfinyl)<Sulfinyl radical>) Imino) -5, 7-dihydrospiro [ cyclopenta [ b ] s]Pyridine-6, 4' -piperidines]-1' -Carboxylic acid ester (3g,7.41mmol) was added to THF (40mL), and BH was slowly added dropwise to the reaction solution under a temperature decrease to 0 deg.C 3(7.5mL,2N in THF), after the addition was complete, the temperature was maintained and the reaction was allowed to proceed for 2 hours. Methanol (10mL) was added dropwise to the reaction. Concentrating under reduced pressure, separating and purifying the obtained crude product by flash silica gel column chromatography to obtain a light yellow solid target compound tert-butyl (S) -5- (((R) -tert-butyl sulfinyl<Sulfinyl radical>) Amino) -5, 7-dihydrospiro [ pentadieno [ b ]]Pyridine-6, 4' -piperidines]-1' -carboxylic acid ester (2.5g, yield 83%).
MS m/z(ESI):408.1[M+H]+.
And a sixth step: preparation of (S) -5, 7-dihydrospiro [ cyclopenta [ b ] pyridine-6, 4' -piperidine ] -5-amine
Figure BDA0003005435270001011
Tert-butyl (S) -5- (((R) -tert-butylsulfinyl)<Sulfinyl radical>) Amino) -5, 7-dihydrospiro [ cyclopenta [ b ] s]Pyridine-6, 4' -piperidines]-1' -Carboxylic acid ester (2.5g,6.14mmol), dioxane hydrochloride (10mL,4N) was added CH2Cl2(40mL), the reaction mixture was stirred at room temperature for 3 hours, and concentrated under reduced pressure to give the desired compound (S) -5, 7-dihydrospiro [ cyclopenta [ b ]]Pyridine-6, 4' -piperidines]-5-amine (1.5g), used directly in the next reaction.
MS m/z(ESI):204.1[M+H]+.
The seventh step: preparation of (S) -1'- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) pyrazin-2-yl) -5, 7-dihydrospiro [ cyclopenta [ b ] pyridine-6, 4' -piperidine ] -5-amine
Figure BDA0003005435270001012
(S) -5, 7-dihydrospiro [ cyclopenta [ b ] pyridine-6, 4' -piperidine ] -5-amine (1.5g,7.39mmol), 3- ((2-amino-3-chloropyridin-4-yl) thio) -6-bromopyrazin-2-amine (3.7g,11.09mmol) and cesium carbonate (7.2g,22.17mmol) were added to DMF (50mL) and reacted at 90 ℃ for 13 hours. The reaction solution was concentrated under reduced pressure, and the crude product was separated and purified by flash column chromatography on silica gel to give (S) -1'- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) pyrazin-2-yl) -5, 7-dihydrospiro [ cyclopenta [ b ] pyridine-6, 4' -piperidine ] -5-amine (1g, yield 29%) as a pale yellow solid.
MS m/z(ESI):455.1[M+H]+,457.1[M+H+2]+.
Eighth step: preparation of (S) -1'- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) -3-fluoropyrazin-2-yl) -5, 7-dihydrospiro [ cyclopenta [ b ] pyridine-6, 4' -piperidin ] -5-amine
Figure BDA0003005435270001013
(S) -1'- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) pyrazin-2-yl) -5, 7-dihydrospiro [ cyclopenta [ b ] pyridine-6, 4' -piperidine ] -5-amine (600mg,1.32mmol), NFSI (830mg,2.64mmol) was added to THF (10 mL). The reaction mixture was reacted at room temperature for 3 hours, concentrated under reduced pressure, and the resulting crude product was separated and purified by flash silica gel column chromatography to give (S) -1'- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) -3-fluoropyrazin-2-yl) -5, 7-dihydrospiro [ cyclopenta [ b ] pyridine-6, 4' -piperidin ] -5-amine (90mg, yield 14%) as a white solid.
MS m/z(ESI):473.0[M+H]+,475.1[M+H+2]+.
1H NMR(400MHz,MeOH-d4)δ8.33(d,J＝4.8Hz,1H),7.82(d,J＝7.2Hz,1H),7.62(d,J＝5.6Hz,1H),7.28-7.25(m,1H),6.01(d,J＝5.6Hz,1H),4.35-4.31(m,2H),4.03(s,1H),3.36-3.31(m,2H),3.25-3.21(m,1H),2.92(d,J＝16.8Hz,1H),1.98-1.84(m,2H),1.64-1.60(m,1H),1.42-1.39(m,1H).
Example 55
Preparation of (S) -1'- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) -3-fluoropyrazin-2-yl) -4, 6-dihydrospiro [ cyclopenta [ d ] thiazole-5, 4' -piperidine ] -6-amine
Figure BDA0003005435270001021
The first step is as follows: preparation of (R) -N- ((S) -1'- (6-amino-5- ((3-chloro-2-cyclopropylpyridin-4-yl) thio) pyrazin-2-yl) -1, 3-dihydrospiro [ indene-2, 4' -piperidin ] -1-yl) -2-methylpropane-2-sulfinamide
Figure BDA0003005435270001022
Reacting (R) -N- ((S) -4, 6-dihydrospiro [ cyclopenta [ d ]]Thiazole-5, 4' -piperidines]-6-yl) -2-methylpropane-2-sulfinamide (260mg, 0.83mmol, 1.0eq), 3- ((2-amino-3-chloropyridin-4-yl) thio) -6-chloropyrazin-2-amine (227mg,0.79mmol,0.95eq) and potassium carbonate (574mg,4.15mmol,5.0eq) were dissolved in DMF (2 mL). The mixture was heated to 105 ℃ under nitrogen protection, and the reaction was stirred for 10 hours. After the reaction is finished, DMF is removed by concentration, and the crude product is subjected to column chromatography (8% MeOH in CH) 2Cl2) To obtain a yellow solid product (R) -N- ((S) -1'- (6-amino-5- ((3-chloro-2-cyclopropylpyridin-4-yl) thio) pyrazin-2-yl) -1, 3-dihydrospiro [ indene-2, 4' -piperidine)]-1-yl) -2-methylpropane-2-sulfinamide (340mg, 76% yield).
MS m/z(ESI):565.1[M+H]+,567.1[M+H+2]+。
The second step is that: preparation of ((S) -1'- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) pyrazin-2-yl) -4, 6-dihydrospiro [ cyclopenta [ d ] thiazole-5, 4' -piperidine ] -6-amine
Figure BDA0003005435270001023
Mixing (R) -N- ((S) -1'- (6-amino-5- ((3-chloro-2-cyclopropylpyridin-4-yl) thio) pyrazin-2-yl) -1, 3-dihydrospiro [ indene-2, 4' -piperidine]-1-yl) -2-methylpropane-2-sulfinamide (340mg, 0.60mmol, 1.0eq) was dissolved in MeOH (5mL), and a solution of HCl in dioxane (1.0mL,4M) was added and reacted at room temperature for 1 hour. The reaction mixture was concentrated to dryness, dissolved in 5mL of methanol, and then dissolved in 7M NH3Adjusting the pH of the methanol solution toBasic, concentrate the reaction, purify it on a normal phase column (10% MeOH in CH)2Cl2) Then purified by reverse phase column (40% MeCN in H)2O(0.1％NH3.H2O) to obtain a grey solid target compound ((S) -1' - (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) pyrazin-2-yl) -4, 6-dihydrospiro [ cyclopenta [ d ]]Thiazole-5, 4' -piperidines]-6-amine (220mg, 80% yield).
MS m/z(ESI):461.1[M+H]+,463.1[M+H+2]+。
The third step: preparation of (S) -1'- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) -3-fluoropyrazin-2-yl) -4, 6-dihydrospiro [ cyclopenta [ d ] thiazole-5, 4' -piperidine ] -6-amine
Figure BDA0003005435270001031
((S) -1' - (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) pyrazin-2-yl) -4, 6-dihydrospiro [ cyclopenta [ d ] thiazole-5, 4' -piperidin ] -6-amine (220mg, 0.48mmol, 1.0eq) was dissolved in anhydrous DMF (2mL) and anhydrous MeCN (10mL) and replaced three times with nitrogen, the reagent N-fluoro-N- (benzenesulfonyl) benzenesulfonamide (159mg,0.5mmol,1.05eq.) was added in portions under nitrogen protection, the mixture was stirred at room temperature for 3 hours, the reaction solution was added dropwise to stirred MeOH (50mL) and then 1mL of water was added, the reaction solution was concentrated to give a crude product which was purified by preparative HPLC to give (S) -1' - (6-amino-5- ((2-amino- 3-Chloropyridin-4-yl) thio) -3-fluoropyrazin-2-yl) -4, 6-dihydrospiro [ cyclopenta [ d ] thiazole-5, 4' -piperidin ] -6-amine (40mg, 17% yield).
1H NMR(400MHz,DMSO)δ8.96(s,1H),7.67(d,J＝5.3Hz,1H),6.28(s,2H),6.17(s,2H),5.86(d,J＝5.4Hz,1H),4.10(s,2H),3.98(s,1H),2.81(dd,J＝52.7,15.2Hz,4H),1.88(s,2H),1.75–1.52(m,4H).
19F NMR(376MHz,DMSO)δ-88.17(s).
MS m/z(ESI):479.0[M+H]+,481.0[M+H+2]+.
Example 56
Preparation of (S) -8- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) -3-fluoropyrazin-2-yl) -2-cyclopropyl-8-azaspiro [4.5] dec-2-en-1-amine
Figure BDA0003005435270001032
The first step is as follows: preparation of (S) -8- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) -3-fluoropyrazin-2-yl) -2-cyclopropyl-8-azaspiro [4.5] dec-2-en-1-amine
Figure BDA0003005435270001041
To a solution of (S) -8- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) pyrazin-2-yl) -2-cyclopropyl-8-azaspiro [4.5] dec-2-en-1-amine (35mg,0.085mmol) in tetrahydrofuran (18mL) at 25 ℃ was added N-fluorobisbenzenesulfonamide (40mg,0.128mmol), and the mixture was stirred for 1 hour. TLC showed no reaction completion, supplemented with N-fluorobisbenzenesulfonamide (27mg,0.085mmol), and stirring was continued for 1 hour. TLC showed no reaction, and was supplemented with N-fluorobisbenzenesulfonamide (27mg,0.085mmol) and stirred for 1 hour. TLC showed the presence of impurities, the reaction was quenched with hydrochloric acid (40mL,0.5N), washed with ethyl acetate (20mL × 2), the aqueous phase was adjusted to pH about 8 with sodium carbonate, extracted with ethyl acetate (30mL × 2), the organic phase was spun off and purified by preparative chromatography (dichloromethane/methanol ═ 10/1) to give (S) -8- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) -3-fluoropyrazin-2-yl) -2-cyclopropyl-8-azaspiro [4.5] dec-2-en-1-amine (28mg, yield: 27%) as a pale yellow solid.
1H NMR(400MHz,MeOD)δ7.61(d,J＝8Hz,1H),5.99(d,J＝8Hz,1H),5.36(s,1H),4.31–4.13(m,2H),3.45(s,1H),3.30–3.15(m,2H),2.43–2.24(m,2H),1.89–1.48(m,5H),0.84–0.65(m,2H),0.57–0.36(m,2H).
MS m/z(ESI):462.1[M+H]+，474.1[M+2+H]+.
Example 57
Preparation of (S) -8- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) -3-fluoropyrazin-2-yl) -2-methyl-8-azaspiro [4.5] dec-2-en-1-amine
Figure BDA0003005435270001042
Example 57 the procedure was as in example 53.
MS m/z(ESI):436.1[M+H]+,438.1[M+2+H]+.
Example 58
Preparation of (S) - (3- (1-amino-2-cyclopropyl-8-azaspiro [4.5] dec-2-en-8-yl) -6- ((2-amino-3-chloropyridin-4-yl) thio) -5-methylpyrazin-2-yl) methanol
Figure BDA0003005435270001043
The procedure for the preparation of example 58 is as described in the experimental scheme of example 45.
MS m/z(ESI):473.1[M+H]+,475.1[M+2+H]+.
Example 59
Preparation of (S) -1'- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) pyrazin-2-yl) -1, 3-dihydrospiro [ indene-2, 4' -piperidine ] -1-amine
Figure BDA0003005435270001051
The procedure for the preparation of example 59 is as described in the experimental scheme of example 53.
1H NMR(400MHz,MeOD)δ7.66–7.54(m,2H),7.42–7.33(m,1H),7.29–7.16(m,3H),5.93(d,J＝5.5Hz,1H),4.37–4.19(m,2H),3.98(s,1H),3.29–3.20(m,2H),3.15(d,J＝16Hz,1H),2.83(d,J＝16Hz,1H),1.91–1.68(m,2H),1.65–1.53(m,1H),1.50–1.39(m,1H).
MS m/z(ESI):454.1[M+H]+,456.1[M+2+H]+.
Second, compound biological test evaluation
The present invention is further described and explained below in conjunction with test examples, which are not intended to limit the scope of the present invention.
Test example 1 determination of the SHP-2 kinase Activity inhibitory Effect of the Compound of the present invention
Purpose of the experiment:
the purpose of this test was to measure the inhibitory ability of compounds on the allosteric activity of the full-length SHP-2 protein.
An experimental instrument: the centrifuge (5810R) is purchased from Eppendorf company, the pipettor is purchased from Eppendorf or Rainin company, and the microplate reader is purchased from BioTek company in the United states and is a SynergyH1 full-function microplate reader.
The experimental method comprises the following steps: in vitro SHP-2 activity assays were performed using the Homogeneous Full Length SHP-2Assay Kit (BPS Bioscience, # 79330). 18 μ L of Master Mix, i.e. 0.5 μ M SHP-2activating Peptide and 5mM DTT in a final concentration of 1 × reaction buffer, was first added to a 96-well low adsorption microplate (NUNC, #267342), 5 μ L of test compound/DMSO per well (final DMSO content 1%, V/V, test compound dissolved in DMSO to 1mM, three-fold serial dilutions were performed, 10 concentrations, reaction system final concentrations ranged from 1 μ M to 0.05nM) was added after centrifugation, SHP-2 was diluted to a final concentration of 0.06nM in 1 × reaction buffer, 2 μ L per well was added to the reaction microplate, full activity control (compound plus DMSO only) and full inhibition control (no SHP-2) were placed on the reaction plate, the reaction mixture was incubated at room temperature for 60 min after centrifugation.
After the incubation was complete, 25. mu.L of Substrate solution containing Substrate at a final concentration of 10. mu.M and 5mM DTT was added to each well and incubation continued for 30 minutes at room temperature after centrifugation. After the reaction is finished, the excitation wavelength is set to be 340nM, the emission wavelength is set to be 455nM, and the gain value is set to be 75 for reading on a Synergy H1 full-function microplate reader (Biotek).
The experimental data processing method comprises the following steps:
percentage inhibition ratio data {% inhibition ratio {% inhibition 100- [ (test compound-Min average)/(Max average-Min average) for wells treated with compound was calculated from the values of total activity control and total inhibition control as Max and Min by positive control wells (DMSO control wells) and negative control wells (no kinase added) on the reaction plate]X 100 }. Using GraphPadprism fitting percent inhibition and ten-point concentration data to a 4-parameter nonlinear logical equation to calculate IC for test compounds50The value is obtained.
And (4) experimental conclusion:
it follows from the above protocol that the compounds of the examples shown in the present invention show biological activities in the SHP-2 kinase activity assay as shown in table 2.1 below,
table 2.1: relative IC of compound for SHP-2 kinase activity inhibition50Value of
Figure BDA0003005435270001061
The above data show that the compounds of the examples shown in the present invention have good activity in the inhibition of SHP-2 kinase activity.
Test example 2 measurement of KYSE520 cell proliferation inhibitory Activity by Compound of the present invention
The purpose of this test example was to determine the inhibitory effect of the compounds of the present invention on the proliferative activity of tumor cells. The tumor cell proliferation inhibiting activity of the compound is measured by a method of CellTiter-Glo, and the half inhibition concentration IC of the compound for inhibiting the cell proliferation activity is obtained 50. 2000 cells per well of a 96-well cell culture plate (Corning, #3610) were seeded with 90. mu.L of a cell suspension, and the plate was cultured overnight (37 ℃ C., 5% CO) in an incubator2). The next day, 10. mu.L of test compound solution (final DMSO content 0.1%, V/V, final reaction concentration range from 100. mu.M to 45nM) was added to the plate cells in a gradient dilution. At the same time, a full-activity control (compound DMSO only) and a full-inhibition control (no cells) were placed on the reaction plate, and the plate was incubated in an incubator for 3 days (37 ℃, 5% CO)2)。
After the incubation, 50. mu.L of CellTiter-Glo reagent was added to each well of the cell culture plate and allowed to stand at room temperature for 10 minutes. The chemiluminescent signal values were read on a Synergy H1 full-function microplate reader (Biotek). The percent inhibition ratio data {% inhibition for compound-treated wells was calculated based on the values of full activity control and full inhibition control as Max and MinRate of production 100- [ (test compound-Min average)/(Max average-Min average)]X 100 }. IC of test compounds was calculated using GraphPad prism to fit the percent inhibition and 8-point concentration data to a 4-parameter nonlinear logistic formula50The value is obtained.
The scheme is as follows: the compounds of the present invention showed about 0.1nM to 200nM (IC) in the KYSE520 cell proliferation inhibitory activity assay 50) The biological activity of (1).
In some embodiments, the compounds of the present invention have IC for KYSE520 cell proliferation inhibitory activity50Less than about 200nM, preferably less than about 100nM, more preferably less than about 10nM, and even more preferably less than about 1nM, and most preferably less than 1nM of the compounds listed in the present invention.
Test example 3 measurement of inhibitory Activity of the Compound of the present invention on NCI-H358 cell proliferation
Purpose of the experiment: the purpose of this test example was to determine the inhibitory effect of the compounds of the present invention on the proliferative activity of tumor cells.
An experimental instrument: the centrifuge (5810R) was purchased from Eppendorf, the carbon dioxide incubator from Thermo, the biosafety cabinet from Shanghai Bowen, the pipette from Eppendorf or Rainin, and the microplate reader from BioTek, USA, model SynergyH1 full-function microplate reader.
The experimental method comprises the following steps: 3000 cells per well of a cell culture plate (Corning, #3610) were seeded with 90. mu.L of a cell suspension, and the plate was cultured overnight (37 ℃ C., 5% CO) in an incubator2). The next day, a solution of test compound diluted in a gradient (final DMSO content 0.2%, V/V, final concentration range of reaction from 10. mu.M to 4.5nM) was added to the plate cells at 10. mu.L per well. At the same time, a full-activity control (compound DMSO only) and a full-inhibition control (no cells) were placed on the reaction plate, and the plate was incubated in an incubator for 3 days (37 ℃, 5% CO) 2)。
After the incubation, 50. mu.L of CellTiter-Glo reagent was added to each well of the cell culture plate and allowed to stand at room temperature for 10 minutes. The chemiluminescent signal values were read on a Synergy H1 full-function microplate reader (Biotek). According to full activity controls andtotal inhibition control as a numerical value for Max and Min, the percent inhibition ratio data {% inhibition 100- [ (test compound-Min average)/(Max average-Min average) for compound-treated wells was calculated]X 100 }. IC of test compounds was calculated using GraphPad prism to fit the percent inhibition and 8-point concentration data to a 4-parameter nonlinear logistic formula50The value is obtained.
The experimental data processing method comprises the following steps:
the percent inhibition data {% inhibition of wells treated with example compound was calculated by calculating the percent inhibition on the plate from the vehicle control wells as 100- (test compound value/vehicle control value) × 100 }. IC was calculated using GraphPad prism to fit different concentrations and corresponding percent inhibition data to a 4-parameter nonlinear logistic formula50The value is obtained.
And (4) experimental conclusion:
the above protocol shows that the compounds of the examples shown in the present invention show biological activities as shown in table 2.2 below in the activity test of proliferation inhibition of NCI-H358 cells.
Table 2.2: IC of Compounds on inhibitory Activity of NCI-H358 cell proliferation 50Value of
Figure BDA0003005435270001081
The above data show that the compounds of the examples shown in the present invention have good activity in the NCI-H358 cell proliferation inhibitory activity.
Test example 4 measurement of mouse pharmacokinetics
4.1. The research aims are as follows:
the pharmacokinetic behavior of the compounds example 36, example 46, example 53, example 54 and example 55 in vivo (plasma) of mice administered orally was investigated using Balb/c mice as test animals.
4.2. Test protocol
4.2.1 test drugs:
embodiment 36, embodiment 46, embodiment 53, embodiment 54 and embodiment 55 of the present invention, self-made;
4.2.2 test animals:
balb/c mice, male, purchased from Shanghai Jitsie laboratory animals Ltd, animal production license number (SCXK (Shanghai) 2013-.
4.2.3 preparation of the medicine:
5g of hydroxyethyl cellulose (HEC, CMC-Na, viscosity: 800-1200Cps) was weighed, dissolved in 1000mL of purified water, and 10g of Tween80 was added. Mix well to get a clear solution.
Example 36, example 46, example 53, example 54 and example 55 were weighed to 1.2mg, added to a 4-mL glass vial, and 2.4mL of the solution was added and sonicated for 10 minutes to give a colorless clear solution at a concentration of 0.5 mg/mL.
4.2.4 dosing:
Balb/c mice, male; after fasting overnight, the oral administration is carried out respectively, the dosage is 5mg/kg, and the administration volume is 10 mL/kg.
4.2.5 sample Collection:
collecting blood before administration and after administration for 0.25h, 0.5h, 1h, 2h, 4h, 6h, 8h, and 24h, placing the blood in EDTA-2K test tube, centrifuging at 4 deg.C and 6000rpm for 6min to separate blood plasma, and storing at minus 80 deg.C; food was consumed 4h after dosing.
4.2.6 measurement results:
the final results obtained by applying LCMS/MS method are shown in Table 2.3
Table 2.3: mouse pharmacokinetic parameters of Compounds
Figure BDA0003005435270001091
The above data show that: at a dose of 5mg/kg, the compounds of the examples of the present invention showed good metabolic activity.
Test example 5 pharmacokinetic assay in rats
5.1. The research aims are as follows:
the pharmacokinetic behavior of example 36, example 46, example 53, example 54, example 55 and example 59 in rats administered orally was investigated using SD rats as test animals.
5.2. Test protocol
5.2.1 test drugs:
examples 36, 46, 53, 54, 55 and 59 of the present invention were prepared by oneself.
5.2.2 test animals:
SD rats were 3 per group, male. Shanghai Jiesi laboratory animals Ltd, animal production license number (SCXK (Shanghai) 2013-.
5.2.3 preparation of the medicine:
5g of hydroxyethyl cellulose (HEC, CMC-Na, viscosity: 800-1200Cps) was weighed, dissolved in 1000mL of purified water, and 10g of Tween80 was added. Mix well to get a clear solution.
3.9mg of example 36, example 46, example 53, example 54, example 55 and example 59 were weighed out, dissolved in the solution, shaken and sonicated for 15 minutes to give a colorless clear solution with a concentration of 0.5 mg/mL.
5.2.4 dosing:
SD rats were administered orally to 3 rats per group in a dose of 5mg/kg in a volume of 10mL/kg in males after fasting overnight.
5.2.5 sample Collection:
collecting 0.2mL of blood from jugular vein before administration and after administration for 0.5h,1.0h,2.0h,4.0h,6.0h,8.0h and 24.0h, placing in EDTA-2K test tube, centrifuging at 4 deg.C and 6000rpm for 6min to separate blood plasma, and storing at-80 deg.C; food was consumed 4h after dosing.
5.3 Experimental results: the final measurements were obtained by applying LCMS/MS method, see Table 2.4
Table 2.4: rat pharmacokinetic parameters of Compounds of the invention
Figure BDA0003005435270001101
And (4) experimental conclusion: the data in the table show that the compounds of the examples of the present invention all achieve relatively high exposure in rat plasma at an orally administered dose of 5 mg/kg; in addition, perianal contamination was observed in rat pharmacokinetic experiments for example 59, indicating that example 59 is gastrointestinal toxic.
Test example 6 inhibition experiment of tumor in MiaPaca 2 transplantation tumor model
6.1 purpose of experiment:
BALB/c nude mice are used as tested animals, and in vivo efficacy experiments are carried out by adopting a human pancreatic cancer cell MiaPaca 2 xenograft tumor (CDX) model to evaluate the anti-tumor effect of tested compounds.
6.2 laboratory instruments and reagents:
6.2.1 Instrument:
super clean bench (BSC-1300II A2, Shanghai Bocheng industry Co., Ltd.)
CO2Incubator (Thermo-311, Thermo)
Centrifuge (Centrifuge 5720R, Eppendorf)
Full-automatic cell counter (Countess II, Life Technologies)
Pipettor (10-20 μ L, Eppendorf)
Microscope (Ts 2, Nikang)
Slide measure (CD-6' AX, Japan Sanfeng)
Cell culture bottles (T25/T75/T225, Corning)
Constant temperature water tank (HWS12, Shanghai-Heng science)
6.2.2 reagents:
DMEM(11995-065，Gibco)
fetal Bovine Serum (FBS) (10091-148, Gibco)
0.25% trypsin (25200-056, Gibco)
Penicillin streptomycin double antibody (P/S) (SV30010, GE)
Phosphate Buffered Saline (PBS) (10010-023, Gibco)
Matrigel(356234，Corning)
Gln(25030-081，Gibco)
6.3 Experimental procedures:
removing MiaPaca 2 cells from cell bank, adding DMEM medium (containing 10% FBS, 1% Glu and 1% P/S) after recovery, and placing in CO2Culturing in incubator (incubator temperature 37 deg.C, CO)2 Concentration 5%). Transferring after the cells are spread to 80-90% of the bottom of the culture bottle Passage, the cells are placed in CO2Culturing in an incubator. Repeating the process until the cell number meets the in vivo drug effect inoculation demand, collecting cells in logarithmic growth phase, counting with a full-automatic cell counter, re-suspending the cells with PBS and Matrigel (volume ratio of 1: 1) according to the counting result, and making into cell suspension (density of 8 × 10)7Ml) and put in an ice box for standby.
Animals used were BALB/c nude mice, female, 6-8 weeks old, and approximately 18-22 grams in weight. Mice were kept in a special pathogen free environment and in a single ventilated cage, 5 mice per cage. All cages, bedding and water were sterilized prior to use and all animals were free to obtain standard certified commercial laboratory diets. The nude mice were marked with disposable universal ear tags for both small and large mice before the start of the experiment, the skin of the inoculated part was disinfected with 75% medical alcohol before the inoculation, and each mouse was subcutaneously inoculated with 0.1ml (containing 8 x 10) of the right back6Individual cells) MiaPaca2 tumor cells. When the average tumor volume reaches 100-3The grouped administration is started. The test compound was administered by oral gavage daily, and the dose, frequency of administration and the drug effect at the end of the experiment are shown in table 5. Tumor volume (mm) was measured twice weekly using a vernier caliper 3) The calculation formula is as follows: v0.5 x D, wherein D and D are the long and short diameters of the tumor, respectively. The anti-tumor efficacy was determined by dividing the mean tumor gain volume of compound-treated animals by the mean tumor gain volume of untreated animals. The tumor inhibition rate is calculated by the formula: TGI (%) ═ 1- [ (Vt-V0) administration group/(Vt-V0) solvent control group]100%. All animals were euthanized at the end of the experiment.
6.4 Experimental results:
table 2.5: transplanted tumor of compound mouse pharmacodynamic parameters
Figure BDA0003005435270001121
Note: the data in parentheses indicate that this example corresponds to the tumor volume of the Vehicle QD x 3w group (i.e., the control group) at the corresponding time (i.e., day 0 and day 21, respectively)
And (4) experimental conclusion: the data show that after oral administration for 21 days, the compound of the embodiment of the invention can obviously inhibit the growth of MiaPaca 2 nude mouse transplanted tumor under the QD administration condition of 3-10 mg/kg.
Test example 7 pharmacokinetic assay of tumor-bearing mice
7.1. The research aims are as follows:
using MiaPaca 2 tumor-bearing mice as test animals, the pharmacokinetic behavior of compound example compound 53 administered orally at a dose of 6mg/kg in vivo (plasma, tumor tissue and intestinal tract) in mice was studied.
7.2. Test protocol
7.2.1 test drugs:
Compound 53, an example of the present invention, was made by house.
7.2.2 test animals:
MiaPaca 2 tumor-bearing mice 24, females. 3 per time point (0h, 1h, 2h, 4h, 6h, 8h, 16h, 24 h). Shanghai Sphere-BiKai laboratory animals Ltd, animal production license number (SCXK (Shanghai) 2018-.
7.2.3 preparation of the medicine:
5g of hydroxymethyl cellulose is weighed out, dissolved in 1000mL of purified water, and 10g of Tween80 is added. Mix well to get a clear solution.
4.57mg of example compound 53, 22.6mg were weighed out and dissolved in this solution, shaken up and sonicated for 15 minutes to give a homogeneous suspension with a concentration of 0.6 mg/mL.
7.2.4 administration:
after fasting, MiaPaca 2 tumor-bearing mice were dosed at 6mg/kg by weight p.o. (animals were not dosed at 0 h) and at 10 mL/kg.
7.2.5 sample collection:
before and after administration to mice, CO2Killing, collecting blood of 0.5ml from heart, placing in EDTA-2K test tube, centrifuging at 4 deg.C and 6000rpm for 6min to separate blood plasma, and storing at-80 deg.C; tumor tissues were weighed, placed in 2mL centrifuge tubes, and stored at-80 ℃. Cutting appropriate length of duodenum, ileum and colon tissue with scissors, removing content, washing with PBS for 2 times, sucking water with absorbent paper, weighing, placing in 2mL centrifuge tube, and sealing Storing at-80 deg.C.
7.3 Experimental results: the final assay results were obtained using LCMS/MS method, see Table 2.6:
table 2.6: mouse pharmacokinetic parameters of Compounds of the invention
Figure BDA0003005435270001131
And (4) experimental conclusion: as shown by the data in the table, the exposure of the compound of the example of the invention to the tumor of the mouse reaches a high level at the dose of 6mg/kg, and the exposure to the tumor is obviously higher than that to the blood. As can be seen from Tmax and MRT, the concentration of the compound in the tumor is a gradually increasing process, and the longer T1/2 indicates that the compound is gradually accumulated in the tumor and always keeps higher concentration in the tumor, thereby ensuring better tumor inhibition effect.
Test example 8 hERG Potassium channel inhibitory Activity test
8.1 cell preparation
8.1.1CHO-hERG cells cultured at 175cm2And (3) in a culture flask, removing the culture solution when the cell density reaches 60-80%, washing once with 7mL of PBS, and adding 3mL of Detachin for digestion.
8.1.2 adding 7mL of culture solution for neutralization after complete digestion, then centrifuging, sucking the supernatant, and adding 5mL of culture solution for heavy suspension to ensure that the cell density is 2-5 multiplied by 106/mL。
8.2 preparation of solution
Table 2.7: composition of intracellular and extracellular fluids
Figure BDA0003005435270001141
8.3 electrophysiological recording procedure
The single cell high impedance sealing and the whole cell mode forming process are all automatically completed by a Qpatch instrument, after a whole cell recording mode is obtained, the cell is clamped at minus 80 millivolts, before a depolarization stimulation of plus 40 millivolts for 5 seconds is given, a 50 millisecond-50 millivolt pre-voltage is given, then repolarization is carried out to minus 50 millivolts for maintaining for 5 seconds, and then the voltage returns to minus 80 millivolts. This voltage stimulus was applied every 15 seconds, and 2 minutes after recording extracellular fluid was administered for 5 minutes, and then the dosing process was started, with compound concentrations starting from the lowest test concentration, each test concentration being administered for 2.5 minutes, and after all concentrations were administered in succession, the positive control compound 3M Cisapride was administered. At least 3 cells (n.gtoreq.3) were tested per concentration.
8.4 preparation of Compounds
8.4.1 the 20mM stock of compound was diluted with the extracellular fluid, 5. mu.L of 20mM stock of compound was added to 2495. mu.L of the extracellular fluid, and 500-fold dilution was carried out to 40. mu.M, followed by 3-fold serial dilutions in the extracellular fluid containing 0.2% DMSO in order to obtain the final concentration to be tested.
8.4.2 the highest concentration tested was 40. mu.M, in turn 6 concentrations of 40, 13.33, 4.44, 1.48, 0.49, 0.16. mu.M.
8.4.3 final test concentration of DMSO content does not exceed 0.2%, the concentration of DMSO on the hERG potassium channel has no effect.
8.5 data analysis
Experimental data were analyzed by XLFit software.
8.6 quality control
Environment: humidity is 20-50%, and temperature is 22-25 DEG C
Reagent: the experimental reagent is purchased from Sigma, and has purity of 98%
The experimental data in the report must meet the following criteria:
whole cell sealing impedance >100M omega
Tail current amplitude >400pA
Pharmacological parameters:
the inhibitory effect of multiple concentrations of Cisapride on the hERG channel was set as a positive control.
8.7 results of the experiment
Results of inhibition of hERG current at multiple concentrations by the compounds of examples:
table 2.8: example inhibition of hERG Current at multiple concentrations
Example numbering hERG(uM)
Example 6 5.24
Example 35 6.17
Example 36 2.34
Example 46 18.78
Example 53 4.51
Example 55 21.91
And (4) experimental conclusion: inhibition of cardiac hERG potassium channel by drugs is the major cause of QT prolongation syndrome by drugs. The experimental results show that the compound provided by the embodiment of the invention has no obvious inhibition effect on the cardiac hERG potassium ion channel, and can avoid the toxic and side effects of the heart at high dose.
Test example 9 measurement of mouse pharmacokinetics
9.1. The research aims are as follows:
balb/c Mouse was used as a test animal to study the pharmacokinetic behavior of the compounds example 53 and example 59 in Mouse (plasma) with a single intravenous administration.
9.2. Test protocol
9.2.1 test drugs:
example 53 and example 59 of the present invention, homemade;
9.2.2 test animals:
balb/c mice, male, purchased from Shanghai Jitsie laboratory animals Ltd, animal production license number (SCXK (Shanghai) 2013-.
9.2.3 preparation of medicine:
10% Solutol HS15 in water: adding purified water for injection into Solutol HS 1510 g, stirring to dissolve, diluting with purified water for injection to 100mL, and shaking to obtain the final product;
20% HP-beta-CD in water: taking 20g of HP-beta-CD, adding purified water for injection, stirring to dissolve, diluting to 100mL with purified water for injection, and shaking up to obtain the compound preparation;
mix 0.5mL of DMSO, 8.5mL of 20% HP- β -CD aqueous solution and 1mL of 10% Solutol HS15 aqueous solution and shake to give a clear solution.
0.234mg of example 53 and example 59 was weighed, 1.17mL of the solution was added thereto, and dissolved by sonication to obtain a clear solution, which was filtered through a 0.22 μm aqueous filter to obtain a filtrate as an iv preparation. The concentration was 0.2 mg/mL.
9.2.4 dosing:
balb/c Mouse, male; after fasting overnight, the drugs were administered separately by intravenous injection at a dose of 1mg/kg and at a volume of 5 mL/kg.
9.2.5 sample Collection:
collecting blood before administration and after administration for 0min,5min, 15min,0.5h,1.0h,2.0h,4.0h,8.0h and 24.0h, placing the blood in EDTA-2K test tube, centrifuging at 4 deg.C and 6000rpm for 6min to separate blood plasma, and storing at minus 80 deg.C; food was consumed 4h after dosing.
9.3 results of the experiment: the final results obtained by applying LCMS/MS method are shown in Table 2.9
Table 2.9: mouse pharmacokinetic parameters of Compounds
Figure BDA0003005435270001161
And (4) experimental conclusion: at a dose of 1mg/kg, the compounds of the examples of the invention showed good metabolic profile in mice.
Test example 10 pharmacokinetic measurement in rat
10.1. The research aims are as follows:
the pharmacokinetic behavior of example 53 and example 59, a single intravenous administration, in vivo (plasma) in rats, was studied using SD rats as test animals.
10.2. Test protocol
10.2.1 test drugs:
inventive examples 53 and 59, self-made.
10.2.2 test animals:
SD rats were 3 per group, male. Shanghai Jiesi laboratory animals Ltd, animal production license number (SCXK (Shanghai) 2013-.
10.2.3 preparation of medicine:
10% Solutol HS15 in water: adding purified water for injection into Solutol HS 1510 g, stirring to dissolve, diluting with purified water for injection to 100mL, and shaking to obtain the final product;
20% HP-beta-CD in water: taking 20g of HP-beta-CD, adding purified water for injection, stirring to dissolve, diluting to 100mL with purified water for injection, and shaking up to obtain the compound preparation;
mix 0.5mL of DMSO, 8.5mL of 20% HP- β -CD aqueous solution and 1mL of 10% Solutol HS15 aqueous solution and shake to give a clear solution.
0.78mg of each of examples 53 and 59 was weighed, 3.9mL of the solution was added thereto, and dissolved by sonication to obtain a clear solution, which was filtered through a 0.22 μm aqueous filter to obtain a filtrate as an iv preparation. The concentration was 0.2 mg/mL.
10.2.4 administration:
SD rats were administered to 3 rats per group by intravenous injection at a dose of 1mg/kg and a volume of 5mL/kg after fasting overnight.
10.2.5 sample collection:
collecting 0.2mL of blood from jugular vein at 0min,5min,15min,0.5h,1.0h,2.0h,4.0h,8.0h and 24.0h before and after administration, placing in EDTA-2K test tube, centrifuging at 4 deg.C and 6000rpm for 6min to separate blood plasma, and storing at-80 deg.C; food was consumed 4h after dosing.
10.3 results of the experiment: the final measurements were obtained by applying LCMS/MS method, see Table 2.10
Table 2.10: rat pharmacokinetic parameters of Compounds of the invention
Figure BDA0003005435270001171
And (4) experimental conclusion: at a dose of 1mg/kg, the compounds of the examples of the invention showed good metabolic profile in rats.
Test example 11 determination of mouse tolerance
11.1. The research aims are as follows:
balb/c mice were used as test animals to study the tolerance of the compounds of example 53 and example 59, orally administered to male and female mice.
11.2. Test protocol
11.2.1 test drugs:
example 53 and example 59 of the present invention, homemade;
11.2.2 test animals:
balb/c mice, 9 males and 9 females, were purchased from Shanghai Jitsie laboratory animals Ltd, animal production license number (SCXK (Shanghai) 2013-.
11.2.3 preparation of medicine:
5g of hydroxyethyl cellulose (HEC, CMC-Na, viscosity: 800-1200Cps) was weighed, dissolved in 1000mL of purified water, and 10g of Tween80 was added. Mix well to get a clear solution.
41.0mg of example 53 and 25.9mg of example 59 were weighed into 100-mL glass bottles, and 41.0mL and 25.9mL of the solution were added, followed by sonication for 10 minutes to give a colorless clear solution at a concentration of 1 mg/mL.
11.2.4 administration:
balb/c mice, 3 males, 3 females; after fasting overnight, the oral administration is carried out respectively, the dosage is 10mg/kg, and the administration volume is 10 mL/kg.
11.2.5 data Collection:
the body weight of the mice was weighed before and after administration every day, and abnormal reactions were observed; food was consumed 4h after dosing.
11.2.6 measurement results:
the final results obtained by weighing with a balance are shown in Table 2.11.
Table 2.11: comparison of body weight changes in mice
Figure BDA0003005435270001181
And (4) experimental conclusion: the data show that the compound of the embodiment of the invention shows good mouse tolerance under the dosage of 10mg/kg, while the compound of the embodiment 59 shows the characteristics of loose stool, hematochezia, obvious weight reduction and the like and has poor tolerance.
In conclusion, the present invention provides a series of highly active, highly selective SHP2 kinase inhibitors having novel structures; the compound shows good pharmacokinetic properties on rats and mice, and shows good drug effect on a Miapaca2 tumor-bearing mouse model; has great potential to be developed into single medicine or combined medicine aiming at tumor diseases.
Research on salt form and crystal form of (S) -8- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) pyrazin-2-yl) -2-cyclopropyl-8-azaspiro [4.5] dec-2-en-1-amine
1.1 Experimental instruments
1.1.1 some parameters of the physicochemical measuring instrument
Figure BDA0003005435270001182
Figure BDA0003005435270001191
1.2 Instrument and conditions for liquid phase analysis
1.2.1 instruments and devices
Name of instrument Model number
Analytical balance Sartorius BSA224S-CW
Water purifier Milli-Q Plus，Millipore
High performance liquid chromatograph Agilent1260
Pump and method of operating the same Agilent G1311B
Sample injector G1329B
Column oven G1316A
Detector G1315D
1.2.2 chromatographic conditions
A chromatographic column: BONUS-RP, 3.5 μm, 4.6 x 75mm
Flow rate: 1mL/min
Column temperature: 40 deg.C
Detection wavelength: 275nm
Sample introduction volume: 5.0. mu.L
Operating time: 15min
Diluent agent: ACN-Water (v/v, 1:1)
Mobile phase: a: water (0.05% trifluoroacetic acid); b: acetonitrile (0.05% trifluoroacetic acid)
T(min) A(％) B(％)
0.00 95 5
10.00 60 40
13.00 20 80
12.01 90 10
15.00 90 10
1.2.3 chromatographic conditions
A chromatographic column: waters Xbridge C18(4.6mm 150mm,3.5 μm)
Flow rate: 1mL/min
Column temperature: 40 deg.C
Detection wavelength: 275nm
Sample introduction volume: 5.0. mu.L
Operating time: 30min
Diluent agent: DMSO (dimethylsulfoxide)
Mobile phase: a: water (0.05% trifluoroacetic acid); b: acetonitrile (0.05% trifluoroacetic acid)
T(min) A(％) B(％)
0.00 90 10
15.00 70 30
25.00 30 70
25.01 90 10
30.00 90 10
2. Preparation of the Compound of formula (V) free base in different crystal forms
Preparation of crystal form A
5g of (S) -2-cyclopropyl-8-azaspiro [4.5 ]]Adding deca-2-en-1-amine hydrochloride into 75mL of NMP, protecting with nitrogen, and adjusting the temperature to 20-30 ℃; 3.17g NaHCO was added3Stirring for 10 min; 4.35g of 3- ((2-amino-3-chloropyridin-4-yl) thio) -6-chloropyrazin-2-amine and 5.2g K were added2CO3Bubbling nitrogen for 30 min; heating to 85 ℃, stirring for 8h, cooling to room temperature, and adding 120mL of DCM and 150mL of pure water; stirring, separating liquid, and extracting 120mL of DCM once; the organic phases are combined, washed by pure water for 5 times to remove NMP, and washed by 15 percent sodium chloride aqueous solution for one time, wherein each time is 180 mL; adding 0.75g of activated carbon into the DCM phase, and stirring for 1 h; filtering, leaching with 30mL of THF, adding 1g of mercaptosilica gel into the filtrate, stirring for 3 hours, filtering, leaching with 30mL of THF, and concentrating the filtrate to dryness to obtain 9g of yellow oily solid; adding 30mL (EA/MTBE-1/1), heating to 50 ℃, pulping for 0.5h, naturally cooling to 20 ℃, stirring for 4h, filtering, leaching MTBE, and vacuum drying to obtain 3.6g of light yellow solid. It has the following XRPD pattern shown in figure 19 and DSC pattern shown in figure 20 after detection and analysis.
Preparation of crystal form B
About 20mg of free base form a was weighed, added with 0.6mL of 2-methyl-tetrahydrofuran and heated to 70 ℃ to dissolve the solid, quickly placed at 2 ℃ and stirred for 12h to precipitate a solid. And finally, quickly centrifuging the solid, removing supernatant, and performing vacuum drying at 40 ℃ to obtain the crystal form B. It has the following XRPD pattern shown in figure 21 and DSC pattern shown in figure 22 through detection and analysis.
Preparation of the third crystal form C
Weighing about 20mg of free base crystal form A into a 2mL glass bottle, adding 200 mu L of LNMP, stirring at room temperature to dissolve the crystal clear, then adding 0.6mL of isopropyl ether to be turbid, stirring at room temperature for 12h, separating out more solids, finally quickly centrifuging the solids, removing supernatant, and drying in vacuum at 40 ℃ to obtain the crystal form C. It has the following XRPD pattern as shown in figure 23 and DSC pattern as shown in figure 24.
3. Screening of compound salt crystal form
3.1 screening of Compound salt form
3.1.1 Experimental purposes:
different counter-ionic acids are selected and by suitable crystallization methods, it is possible to detect which counter-ionic acids can form the compound salt.
3.1.2 Experimental procedures:
1) apparatus and device
Name (R) Model number Source
Analytical balance BSA224S-CW Sartorius
Ultrasonic cleaning instrument SK5200LHC Shanghai department leads ultrasonic instrument
Liquid-transfering gun Eppendorf(50mL,1000μL) Eppendorf
2) Operating procedure
Firstly, a natural volatilizing method of a mixed solvent of methanol and ethanol is adopted
Weighing 260mg of free base (amorphous), adding 3.4mL of ethanol, adding 1.7mL of methanol, filtering, uniformly dividing the filtered filtrate, adding a certain amount of acid (the molar reaction ratio of the base to the acid is 1: 1.2) into each part, then volatilizing the solvent at room temperature in an open air, and after volatilization, respectively adding 200 mu LEA into an oil-forming sample and a sample with poor crystallinity shown by XRD and pulping. The method comprises the following specific steps:
numbering Acid(s) After addition of acid Result of volatilization Beating result
1 1M hydrochloric acid in EtOH Is still clear Oil of formation Amorphous solid
2 1M p-toluenesulfonic acid in EtOH Is still clear Has solid and poor crystallinity Difference in crystallinity
3 0.25M fumaric acid in EtOH Is still clear Has solid and poor crystallinity Difference in crystallinity
4 1M succinic acid in MeOH Is still clear With solid, viscous sample, uncharacterized Difference in crystallinity
5 / Is still clear Has solid, good crystallinity, and free base -
Results and discussion: the above experiment resulted in a crystalline form of amorphous hydrochloride, p-toluenesulfonate, fumarate, succinate and free base.
② solid-liquid reaction crystallization by adopting Acetone as solvent
10mg of the free base (amorphous) was weighed, 0.1ml of a lactone was added, the mixture was stirred at room temperature and suspended in white, and an acid (base: acid: molar reaction ratio 1: 1.2) was added to the suspension to carry out a reaction, specifically as follows:
Figure BDA0003005435270001211
Figure BDA0003005435270001221
Results and discussion: the above experiments yielded the hydrochloride, phosphate, sulfate and p-toluenesulfonate salts.
Solid-liquid reaction crystallization by adopting EA as solvent
10mg of free base (form a) was weighed, 0.1ml of lea was added, the mixture was suspended with stirring at room temperature, and an acid (base: acid: molar reaction ratio 1: 1.2) was added to the suspension to carry out a reaction as follows:
Figure BDA0003005435270001222
results and discussion: all the above experiments finally gave a solid, forming an acid salt.
Liquid-liquid reaction crystallization (partial stronger acid salt) by adopting THF as solvent
10mg of free base (form A) was weighed, 0.1ml of THF was added, the mixture was stirred at 50 ℃ and dissolved in water, and acid (base: acid: 1: 1.2 molar ratio) was added to the solution to carry out the reaction as follows:
Figure BDA0003005435270001223
results and discussion: the above experiments gave the hydrochloride, isethionate, fumarate and almost amorphous phosphate salts with better crystallization properties.
Adopting ACN as solvent to make solid-liquid reaction crystallization
10mg of free base (form a) was weighed, 0.1ml of lacn was added, the mixture was suspended with stirring at room temperature, and an acid (base: acid: molar reaction ratio 1: 1.2) was added to the suspension to carry out a reaction, as follows:
numbering Acid(s) After addition of acid Post-treatment Results
1 1M hydrochloric acid in EtOH Reaction to form oil and adhere to the wall Stirring overnight at room temperature Better crystallinity
2 1M p-toluenesulfonic acid in EtOH Some sticking to the wall Stirring overnight at room temperature Better crystallinity
3 1M Benzenesulfonic acid in EtOH Quick dissolving, clear, small quantity of insoluble wall adhering Stirring overnight at room temperature Better crystallinity
4 1M Ethanesulfonic acid in EtOH Quick dissolving, clear, small quantity of insoluble wall adhering Stirring overnight at room temperature Better crystallinity
Results and discussion: the experiment results in hydrochloride, p-toluenesulfonate, benzenesulfonate and ethanesulfonate with good crystallinity.
Liquid-liquid reaction crystallization (partial weaker acid salt) using THF as solvent
10mg of free base (form A) was weighed, 0.1ml of THF was added, the mixture was stirred at 50 ℃ and dissolved in water, and acid (base: acid: 1: 1.2 molar ratio) was added to the solution to carry out the reaction as follows:
numbering Acid(s) After addition of acid Post-treatment Results
1 0.5M succinic acid in EtOH Turbidity Stirring overnight at 50 deg.C Good crystallization property
Results and discussion: the succinate obtained by the experiment has good crystallization property.
Seventhly, solid-liquid reaction crystallization (partial stronger acid salt) is carried out by adopting EtOH as solvent
20mg of the free base (form a) were weighed out, 0.1ml of etoh was added, the white suspension was stirred at room temperature, and the acid (base: acid: molar ratio 1: 1.2) was added to the suspension for the reaction, as follows:
numbering Acid(s) After addition of acid Post-treatment Results
1 1M hydrochloric acid in EtOH Remains suspended and turns dark Stirring at room temperature for 2h Better crystallinity
2 1M hydrobromic acid in EtOH Solid is separated out after dissolving and cleaning Stirring at room temperature for 2h Crystallinity is general
3 1M p-toluenesulfonic acid in EtOH Solid is separated out after dissolving and cleaning Stirring at room temperature for 2h Crystallinity is general
Results and discussion: the above experiments yielded the hydrochloride, hydrobromide and p-toluenesulfonate salts.
Liquid-liquid reaction crystal (hydrochloride and phosphate) with THF as solvent
20mg of free base (crystal form A) is weighed, 0.2ml of LTHF is added, the mixture is heated and stirred at 50 ℃ to be dissolved clearly, and acid (the molar reaction ratio of the base to the acid is 1: 1.2) is added into the solution for reaction, and the reaction is carried out as follows:
Figure BDA0003005435270001231
results and discussion: the hydrochloride obtained by the above experiments has consistent crystal form.
Ninthly, adopts EtOH as a solvent to carry out solid-liquid reaction crystallization (part of weaker acid salt)
20mg of the free base (form a) was weighed out, 0.1mL of EtOH was added, the white suspension was stirred at 50 ℃, and the acid (base: acid: molar ratio 1: 1.2) was added to the suspension for reaction as follows:
Figure BDA0003005435270001232
Figure BDA0003005435270001241
results and discussion: the above experiments yielded tartrate and maleate salts.
3.1.3 results of the experiment
Through a salt type screening experiment, 11 salts with crystal forms, namely mesylate, isethionate, fumarate, p-toluenesulfonate, ethanesulfonate, succinate, hydrobromide, hydrochloride, maleate, tartrate and sulfate, are obtained; one crystal form is obtained in the salt screening process and is respectively named as a crystal form A.
4 hygroscopicity test
4.1 purpose of the experiment
And (3) inspecting the hygroscopicity of different salts of the compound under different relative humidity conditions, and providing a basis for screening and storing the salt of the compound.
4.2 protocol:
and (3) placing the compound salt in saturated water vapor with different relative humidity to enable the compound and the water vapor to reach dynamic equilibrium, and calculating the percentage of moisture absorption weight gain of the compound after equilibrium.
4.3 Experimental results:
4.3.1 hygroscopicity of the Compound of formula (V)
1) The mesylate crystal form A has moisture absorption and weight gain of 3.794 percent under the condition of RH80 percent and has moisture absorption. After 1 humidity absorption and desorption circulation under the condition of 0-95% relative humidity, the XRPD spectrogram of the mesylate crystal form A is not changed, namely the crystal form is not transformed.
2) The isethionate salt crystal form A has moisture absorption and weight gain of 7.40% under the condition of RH 80%, and has moisture absorption. And after the moisture absorption and desorption cycle is carried out for 1 time under the condition of 0-95% relative humidity, the XRPD spectrogram of the crystal form A of the isethionate is not changed, namely the crystal form is not transformed.
3) The fumarate salt of the crystal form A absorbs moisture and gains weight 1.819% under the condition of RH 80%, and is slightly hygroscopic. And after moisture absorption and moisture desorption circulation is carried out for 1 time under the condition of 0-95% relative humidity, the XRPD spectrogram of the fumarate crystal form A is not changed, namely the crystal form is not transformed.
4) The p-toluenesulfonate salt form A had a moisture absorption gain of 1.482% under the RH 80% condition, and was slightly hygroscopic. And after the moisture absorption and desorption cycle is carried out for 1 time under the condition of 0-95% relative humidity, the XRPD spectrogram of the p-toluenesulfonate crystal form A is not changed, namely the crystal form is not transformed.
5) The succinate crystal form A has moisture absorption and weight increment of 1.429 percent under the condition of RH80 percent and has hygroscopicity. And after 1 time of moisture absorption and moisture desorption circulation under the condition of 0-95% relative humidity, the XRPD spectrogram of the succinate crystal form A is not changed, namely the crystal form is not transformed.
6) The hydrobromide crystal form A absorbs moisture and gains weight of 4.245% under the condition of RH 80%, and has hygroscopicity. And after moisture absorption and moisture desorption circulation is carried out for 1 time under the condition of 0-95% relative humidity, the XRPD spectrogram of the hydrobromide crystal form A is not changed, namely the crystal form is not transformed.
7) The hydrochloride crystal form A absorbs moisture and gains weight of 3.317 percent under the condition of RH80 percent and has hygroscopicity. And after 1 humidity absorption and desorption circulation under the condition of 0-95% relative humidity, the XRPD spectrogram of the hydrochloride crystal form A is not changed, namely the crystal form is not transformed.
8) The maleate crystal form A has moisture absorption and weight increment of 3.465% under the condition of RH 80% and has hygroscopicity. And after 1 humidity absorption and desorption circulation under the condition of 0-95% relative humidity, the XRPD spectrogram of the maleate crystal form A is not changed, namely the crystal form is not transformed.
5. Solid stability test
5.1 purpose of experiment:
the physical and chemical stability of the compounds of different salt forms at high temperature of 60 ℃, high humidity RH of 92.5 percent and high temperature of 50 ℃ and high humidity RH of 75 percent is considered, and a basis is provided for salt form screening and compound salt storage.
5.2 protocol:
about 2mg of different salts are taken, the high temperature of an oven is 60 ℃, the high humidity RH is 92.5 percent, the high humidity RH is 75 percent, the content of the salts is measured by HPLC and an external standard method after 5 days and 10 days are considered, and the change of related substances of the salts is calculated by a chromatographic peak area normalization method (the chromatographic condition is 1.2.2).
5.3 Experimental results:
Figure BDA0003005435270001251
Figure BDA0003005435270001261
RRT/Area％ 1.00 1.02 1.19 1.24 1.62
hydrochloride salt-0 day 64.40 34.28 <0.05 <0.05 <0.05
60-5 days 64.79 33.73 0.12 0.06 0.06
60-10 days 47.19 51.62 0.11 0.07 <0.05
RH is 92.5% -5 days 65.28 33.39 0.03 0.04 0.05
50 ℃ and RH 75-5 days 64.71 33.95 0.08 0.08 0.05
50 ℃ and RH 75-10 days / / 0.10 / 0.12
Hydrobromide salt-0 day 64.54 33.47 / / 0.10
60-5 days 64.67 33.56 / 0.04 0.09
RH is 92.5% -5 days 64.59 33.42 / 0.04 0.10
50 ℃ and RH 75-5 days 64.61 33.62 / 0.15 0.11
Fumarate salt for-0 day 64.11 34.06 0.03 / 0.09
60-5 days 63.35 34.96 / / 0.08
RH is 92.5% -5 days 64.18 34.23 0.03 / 0.09
50 ℃ and RH 75-5 days 64.36 33.99 0.03 / 0.09
P-toluenesulfonate-0 days 62.68 35.75 / / 0.08
60-5 days 62.73 35.61 / / 0.10
RH is 92.5% -5 days 62.84 35.51 / / 0.08
50 ℃ and RH 75-5 days 62.79 35.55 / / 0.09
5.4 conclusion of the experiment
The diluent used in the liquid phase method can generate solvent effect, so that the main peak is branched into two peaks, and the integration and comparison of the impurity increase conditions show that the fumarate, the p-toluenesulfonate and the succinate are stable under various conditions and have no obvious impurity increase.
6. Solubility experiments in different media
6.1 purpose of the experiment
The solubility of the p-toluenesulfonate crystal form A, the fumarate crystal form A and the succinate crystal form A in media such as different pH values, water, artificial Simulated Gastric Fluid (SGF), fasting artificial simulated intestinal fluid (FaSSIF), non-fasting artificial simulated intestinal fluid (FeSSIF) and the like is compared, and a basis is provided for the evaluation of the druggability of the salt.
6.2 protocol:
approximately 2mg of compound was suspended in various media for 24 hours and thermodynamic solubility of compound at 37 ℃ was determined by HPLC, external standard method.
6.3 Experimental results:
crystalline form A of p-toluenesulfonate Fumarate salt form a Succinate crystal form A
Solvent Solubility mg/mL Solubility mg/mL Solubility mg/mL
Buffer pH 1.0 >1.331 - -
Buffer pH 2.0 >1.24 - -
Buffer pH 3.0 0.495 - -
Buffer pH 4.0 0.385 - -
Buffer pH 5.0 0.203 - -
Buffer pH 6.0 0.724 - -
Buffer pH 7.0 0.525 - -
Buffer pH 8.0 0.069 - -
Water (W) 0.764 0.153 0.214
FaSSGF >1.812 >1.888 >1.803
FaSSIF 0.182 0.114 0.118
FeSSIF 0.303 0.613 0.641
7. Rat PK study of different salt types
7.1 purpose of the experiment
Taking an SD rat as a test animal, researching the pharmacokinetic behaviors of the p-toluenesulfonate crystal form A, the fumarate crystal form A and the succinate crystal form A in vivo (plasma) of the rat after single oral administration, and comparing the change of exposure; and researching the pharmacokinetics of single intravenous administration of the fumarate crystal form A, and calculating the bioavailability of oral administration of the p-toluenesulfonate crystal form A, the fumarate crystal form A and the succinate crystal form A.
7.2 protocol:
after the p-toluenesulfonate crystal form A, the fumarate crystal form A and the succinate crystal form A are uniformly suspended by using an aqueous solution containing 0.5 percent of HPMC K4M, the mixture is subjected to intragastric administration and administration to rats, three rats are treated in parallel, and the administration dose is 10mg/kg of suspension); fumarate crystal form A is dissolved and cleaned by a physiological saline solution containing 20 percent of HP-beta-CD, filtered by a filter membrane, injected into a vein, administered to rats and paralleled to three rats.
7.3 Experimental results:
Figure BDA0003005435270001281
Figure BDA0003005435270001282
8. solid stability test
8.1 purpose of the experiment:
the stability comparison of the p-toluenesulfonate crystal form A, the fumarate crystal form A and the succinate crystal form A under three conditions of high temperature (60 ℃), high humidity at room temperature (RH ═ 92.5%) and high humidity at high temperature (50 ℃), RH ═ 75%) is considered, and a basis is provided for salt type screening and compound salt storage.
8.2 protocol:
taking about 2mg of the p-toluenesulfonate crystal form A, the fumarate crystal form A and the succinate crystal form A, observing for 7 days and 14 days at high temperature (60 ℃), high humidity at room temperature (RH 92.5%) and high humidity (RH 75%) at 50 ℃, measuring the salt content by using HPLC and an external standard method, and calculating the change of related substances of the salt by using a chromatographic peak area normalization method (the chromatographic condition is 1.2.3).
8.3 Experimental results:
Figure BDA0003005435270001283
Figure BDA0003005435270001291
8.4 conclusion of the experiment:
the small impurity increase of p-toluenesulfonate is less than 0.1%.
Salt form and crystal form research of tetra (S) -1'- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) -3-fluoropyrazin-2-yl) -1, 3-dihydrospiro [ indene-2, 4' -piperidine ] -1-amine
Scale-up production of amorphous form of (S) -1'- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) -3-fluoropyrazin-2-yl) -1, 3-dihydrospiro [ indene-2, 4' -piperidin ] -1-amine (example 53)
Adding (S) -1'- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) pyrazin-2-yl) -1, 3-dihydrospiro [ indene-2, 4' -piperidine ] -1-amine and THF into a reaction kettle, adding N-fluoro-diphenyl sulfonamide (NFSI), reacting at room temperature for 2 hours, adding 2L of 0.5mol/L hydrochloric acid solution, adding 2L of EA, stirring and extracting, separating, adding 3L of 0.25mol/L hydrochloric acid solution into the organic phase, adding 11L of EA, stirring and extracting, separating, combining the aqueous phases, discarding the organic phase, adding 1.5L of EA into the aqueous phase, extracting and removing impurities, adding 2.65L of THF into the aqueous phase, stirring, adding sodium carbonate solid to adjust the pH to 9-10, separating, extracting the water phase with 1.5L 2-methyltetrahydrofuran, combining the organic phases, drying, filtering, concentrating to obtain 73.4g yellow brown solid, adding tetrahydrofuran to the solid, and sequentially adding EA: 20% PE, 60%, EA containing 0.005% triethylamine, methanol: the product was rinsed with DCM ═ 5% (5% triethylamine in methanol) and the buffer was concentrated under reduced pressure to give 41.1g of a yellow solid for use.
Study on salt form of Compound
1. Screening for Compound salt form
1.1 purpose of experiment:
different counter-ionic acids are selected and by suitable crystallization methods, it is possible to detect which counter-ionic acids can form the compound salt.
1.2 Experimental procedures:
3) apparatus and device
Name (R) Model number Source
Analytical balance BSA224S-CW Sartorius
Ultrasonic cleaning instrument SK5200LHC Shanghai department leads ultrasonic instrument
Liquid-transfering gun Eppendorf(50mL,1000μL) Eppendorf
2) Operating procedure
Naturally volatilizing ethanol as solvent
160mg of (S) -1'- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) -3-fluoropyrazin-2-yl) -1, 3-dihydrospiro [ indene-2, 4' -piperidine ] -1-amine are weighed out as amorphous, are heated to 50 ℃ with the addition of 3mL of ethanol and completely cleared, the ethanol solution of (S) -1'- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) -3-fluoropyrazin-2-yl) -1, 3-dihydrospiro [ indene-2, 4' -piperidine ] -1-amine is divided on average into 16 parts, a defined amount of acid (molar reaction ratio base: acid 1: 1.2) is added per part, then, the solvent is volatilized at room temperature, oil is formed after volatilization, and 200 mu L of EA is added respectively for pulping. Respectively salifying with hydrochloric acid, sulfuric acid, phosphoric acid, hydrobromic acid, p-toluenesulfonic acid, methanesulfonic acid, benzoic acid, citric acid, fumaric acid, maleic acid, malic acid, succinic acid, tartaric acid, ethanesulfonic acid, oxalic acid and isethionic acid.
② adopting EA as solvent to make liquid-liquid reaction crystallization
10mg of (S) -1'- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) -3-fluoropyrazin-2-yl) -1, 3-dihydrospiro [ indene-2, 4' -piperidine ] -1-amine was weighed out as an amorphous form, 0.2mL of EA was added thereto, the mixture was stirred at room temperature to be clear, and an acid (base: acid: molar reaction ratio of 1: 1.2) was added to the solution to carry out a reaction. Salification is carried out with hydrochloric acid, sulfuric acid, phosphoric acid, hydrobromic acid, p-toluenesulfonic acid, methanesulfonic acid, benzenesulfonic acid, citric acid, fumaric acid, maleic acid, malic acid, succinic acid, tartaric acid, ethanesulfonic acid, oxalic acid and isethionic acid. Thirdly, adopting a natural volatilizing method
About 120mg of (S) -1'- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) -3-fluoropyrazin-2-yl) -1, 3-dihydrospiro [ indene-2, 4' -piperidin ] -1-amine are weighed out as an amorphous, 2.4mL of EtOH, 0.8mL of MeOH are dissolved at room temperature, the filtrate is filtered through a filter, the filtrate is divided equally into 7 parts, the following acids are added, and the solvent is evaporated by exposure to the atmosphere at room temperature. The salt is formed by adopting lactic acid, malonic acid, benzoic acid, adipic acid, hippuric acid, acetic acid and glycollic acid. And fourthly, reacting and crystallizing by using acetonitrile as a solvent, weighing 10mg of (S) -1'- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) -3-fluoropyrazin-2-yl) -1, 3-dihydrospiro [ indene-2, 4' -piperidine ] -1-amine amorphous, adding 0.2mL of ACN, stirring at room temperature until the materials are not completely dissolved, and adding acid (alkali: acid: molar reaction ratio of 1: 1.2) into the system for reaction. Forming salt by using hydrobromic acid, hydroxyethyl sulfonic acid, gentisic acid and fumaric acid.
2. Ethanesulfonate solvent residue investigation
The solvent residues of methanol, ethanol, acetone, methylene chloride, t-butyl methyl ether, ethyl acetate, tetrahydrofuran and 2-methyltetrahydrofuran were measured according to the residual solvent measurement method (third method 0861 in the four parts of the pharmacopoeia 2015 edition), and the results are shown in the following table,
TABLE 4.1 summary of solvent residue detection results
Figure BDA0003005435270001311
And (4) experimental conclusion: the data show that all residual solvents of the ethanesulfonate meet the requirements of national pharmacopoeia standards
4 ethane sulfonate stability study
The ethanesulfonic acid salt of (S) -1'- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) -3-fluoropyrazin-2-yl) -1, 3-dihydrospiro [ indene-2, 4' -piperidine ] -1-amine was weighed approximately 100mg each and tested for stability under different conditions for 5 days and 10 days, respectively, by high performance liquid chromatography (chinese pharmacopoeia 2015 edition, general rules of the four parts 0512) with the following results:
TABLE 4.2 summary of stability findings
Figure BDA0003005435270001312
And (4) experimental conclusion: from the data, the ethane sulfonate has no obvious change in total impurities under the conditions of high temperature, high humidity and illumination for 10 days; in the single hybrid aspect, the RRT is 0.64 under high humidity condition, and the RRT is not changed basically under high temperature condition.
5. Confirmation of salt formation content of hydrochloride
The test method comprises the following steps:
weighing different amounts of NaCl, adding a certain amount of diluent CH3CN:H2O is 1:1 dissolving the sodium chloride in water to obtain NaCl solution with different concentrations, and detecting and analyzing Cl with different concentrations by ELSD-And (4) content. Adding Cl-The concentration is the abscissa, and the corresponding Cl-peak area obtained by ELSD detection is the ordinate to be used as standard curve. In addition, a certain amount of hydrochloride crystal form A is accurately weighed and a certain amount of diluent CH is added3CN:H2O is 1:1 dissolving the mixture in water, detecting and analyzing the obtained solution with ELSD, and obtaining the result according to standard curveAnd weighing the sample amount to accurately calculate Cl-And (4) content.
The analysis method comprises the following steps:
Figure BDA0003005435270001321
as a result:
ESLD detection analysis shows that the mole ratio of free alkali to chloride ion content in hydrochloric acid is 1:1, the salt formation ratio of (S) -1'- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) -3-fluoropyrazin-2-yl) -1, 3-dihydrospiro [ indene-2, 4' -piperidine ] -1-amine free base to hydrochloric acid can be seen to be 1: 1.
(III) study of Crystal form of Compound salt
1. Compound salt crystal form screening
1.1 purpose of experiment:
and selecting a proper crystallization method according to the salt form screening result, and screening different crystal forms of different salts.
1.2 Experimental instruments
1.2.1 some parameters of the physicochemical measuring instrument
Figure BDA0003005435270001331
1.3 Instrument and conditions for liquid phase analysis
1.3.1 instruments and apparatus
Name of instrument Model number
Analytical balance Sartorius BSA224S-CW
Water purifier Milli-Q Plus，Millipore
High performance liquid chromatograph Agilent1260
Pump and method of operating the same Agilent G1311B
Sample injector G1329B
Column oven G1316A
Detector G1315D
1.3.2 chromatographic conditions
A chromatographic column: ZORBAX (SB-C8, 3.5 μm, 4.6X 75mm)
Flow rate: 1mL/min
Column temperature: 40 deg.C
Detection wavelength: 275nm
Sample introduction volume: 5.0. mu.L
Operating time: 13min
Diluent agent: ACN-Water (v/v, 1:1)
Mobile phase: a: water (0.05% trifluoroacetic acid); b: acetonitrile (0.05% trifluoroacetic acid)
Figure BDA0003005435270001332
Figure BDA0003005435270001341
1.4 operating procedure
Preparation of hydrochloride crystal form A of (S) -1'- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) -3-fluoropyrazin-2-yl) -1, 3-dihydrospiro [ indene-2, 4' -piperidine ] -1-amine
Weighing 200mg of (S) -1' - (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) -3-fluoropyrazin-2-yl) -1, 3-dihydrospiro [ indene-2, 4' -piperidine ] -1-amine amorphous, adding 2mL of ethanol, stirring at 50 ℃ to obtain a clear solution, slowly adding 0.467mL of 1M ethanol solution of hydrochloric acid into the system, precipitating a large amount of solid in the acid adding process, stirring at 50 ℃ for 2h, cooling, filtering and drying to obtain (S) -1' - (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) -3-fluoropyrazin-2-yl) -1, 3-dihydrospiro [ indene-2, 4' -piperidine ] -1-amine hydrochloride form a. By detection analysis, the X-ray diffraction data shown in figure 1, the DSC chart shown in figure 2 and the TGA chart shown in figure 3 are obtained.
Preparation of hydrochloride crystal form B of (S) -1'- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) -3-fluoropyrazin-2-yl) -1, 3-dihydrospiro [ indene-2, 4' -piperidine ] -1-amine
Weighing 20mg of (S) -1' - (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) -3-fluoropyrazin-2-yl) -1, 3-dihydrospiro [ indene-2, 4' -piperidine ] -1-amine hydrochloride crystal form A, adding 0.6mL of 88% acetone, heating and dissolving at 75 ℃, then quickly cooling to 0 ℃, stirring for 2h to separate out a solid, finally filtering and drying to obtain (S) -1' - (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) -3-fluoropyrazin-2-yl) -1, 3-dihydrospiro [ indene-2, 4' -piperidine ] -1-amine hydrochloride form B. By detection analysis, the XRPD ray diffraction data shown in the table 2, the DSC chart shown in the figure 4 and the TGA chart shown in the figure 5 are obtained.
Preparation of ethanesulfonate crystal form A of (S) -1'- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) -3-fluoropyrazin-2-yl) -1, 3-dihydrospiro [ indene-2, 4' -piperidine ] -1-amine
Weighing 10mg of (S) -1' - (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) -3-fluoropyrazin-2-yl) -1, 3-dihydrospiro [ indene-2, 4' -piperidine ] -1-amine amorphous, adding 0.1mL of ethyl acetate, stirring at 50 ℃ to obtain a clear solution, slowly adding 0.0255mL of 1M ethane sulfonic acid methanol solution into the system, stirring to obtain oil after adding acid, precipitating a large amount of solid, stirring at 50 ℃ for 2h, cooling, filtering and drying to obtain (S) -1' - (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) -3-fluoropyrazin-2-yl) -1, 3-dihydrospiro [ indene-2, 4' -piperidine ] -1-aminoethanesulfonate crystalline form a. By detection analysis, the XRPD ray diffraction data is shown as figure 6, the DSC chart is shown as figure 7, and the TGA chart is shown as figure 8.
Preparation of besylate crystal form A of (S) -1'- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) -3-fluoropyrazin-2-yl) -1, 3-dihydrospiro [ indene-2, 4' -piperidine ] -1-amine
Weighing 10mg of (S) -1' - (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) -3-fluoropyrazin-2-yl) -1, 3-dihydrospiro [ indene-2, 4' -piperidine ] -1-amine amorphous, adding 0.2mL of ethyl acetate, stirring at 50 ℃ to obtain a clear solution, slowly adding 0.0255mL of 1M benzenesulfonic acid methanol solution into the system, stirring to obtain oil after adding acid, precipitating a large amount of solid, stirring at 50 ℃ for 2h, cooling, filtering and drying to obtain (S) -1' - (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) -3-fluoropyrazin-2-yl) -1, 3-dihydrospiro [ indene-2, 4' -piperidine ] -1-amine benzenesulfonate form a. By detection analysis, the XRPD ray diffraction data shown in the table 3, the DSC chart shown in the figure 9 and the TGA chart shown in the figure 10 are obtained.
Preparation of fumarate crystal form A of (S) -1'- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) -3-fluoropyrazin-2-yl) -1, 3-dihydrospiro [ indene-2, 4' -piperidine ] -1-amine
Weighing 10mg of (S) -1' - (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) -3-fluoropyrazin-2-yl) -1, 3-dihydrospiro [ indene-2, 4' -piperidine ] -1-amine amorphous, adding 0.2mL of ethyl acetate, stirring at 50 ℃ to obtain a clear solution, slowly adding 0.102mL of 0.25M ethanol solution of fumaric acid into the system, stirring to obtain oil after adding the acid, precipitating a large amount of solid, stirring at 50 ℃ for 2h, cooling, filtering and drying to obtain (S) -1' - (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) -3-fluoropyrazin-2-yl) -1, 3-dihydrospiro [ indene-2, 4' -piperidine ] -1-amine fumarate in crystalline form a. By detection analysis, the XRPD ray diffraction data shown in the table 4, the DSC chart shown in the figure 11 and the TGA chart shown in the figure 12 are obtained.
Preparation of hydroxyethyl sulfonate crystal form A of (S) -1'- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) -3-fluoropyrazin-2-yl) -1, 3-dihydrospiro [ indene-2, 4' -piperidine ] -1-amine
Weighing 10mg of (S) -1' - (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) -3-fluoropyrazin-2-yl) -1, 3-dihydrospiro [ indene-2, 4' -piperidine ] -1-amine amorphous, adding 0.2mL of ethyl acetate, stirring at 50 ℃ to obtain a clear solution, slowly adding 0.0255mL of 1M methanol solution of isethionic acid into the system, stirring to obtain oil after adding acid, precipitating a large amount of solid, stirring at 50 ℃ for 2h, cooling, filtering and drying to obtain (S) -1' - (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) -3-fluoropyrazin-2-yl) -1, 3-dihydrospiro [ indene-2, 4' -piperidine ] -1-amine isethionate crystalline form a. By detection analysis, the XRPD ray diffraction data shown in the table 5, the DSC chart shown in the figure 13 and the TGA chart shown in the figure 14 are obtained.
(S) -1'- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) -3-fluoropyrazin-2-yl) -1, 3-dihydrospiro [ indene-2, 4' -piperidine ] -1-amine oxalate crystalline form A preparation
Weighing 10mg of (S) -1' - (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) -3-fluoropyrazin-2-yl) -1, 3-dihydrospiro [ indene-2, 4' -piperidine ] -1-amine amorphous, adding 0.2mL of ethyl acetate, stirring at 50 ℃ to obtain a clear solution, slowly adding 0.0255mL of 1M oxalic acid methanol solution into the system, stirring to separate out a large amount of solid after adding the acid, stirring at 50 ℃ for 2h, cooling, filtering and drying to obtain (S) -1' - (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) -3-fluoropyrazin-2-yl) -1, 3-dihydrospiro [ indene-2, 4' -piperidine ] -1-amine oxalate form a. By detection analysis, the XRPD ray diffraction data shown in the table 6, the DSC chart shown in the figure 15 and the TGA chart shown in the figure 16 are obtained.
Preparation of crystalline form A of the hydrobromide of (S) -1'- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) -3-fluoropyrazin-2-yl) -1, 3-dihydrospiro [ indene-2, 4' -piperidine ] -1-amine
Weighing 20mg of (S) -1' - (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) -3-fluoropyrazin-2-yl) -1, 3-dihydrospiro [ indene-2, 4' -piperidine ] -1-amine amorphous, adding 0.2mL of ethyl acetate, stirring at 50 ℃ to obtain a clear solution, slowly adding 0.04mL of 1M ethanol solution of hydrobromic acid into the system, stirring to separate out a large amount of solid after adding acid, stirring at 50 ℃ for 2h, cooling, filtering and drying to obtain (S) -1' - (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) -3-fluoropyrazin-2-yl) -1, 3-dihydrospiro [ indene-2, 4' -piperidine ] -1-amine hydrobromide crystalline form a. Upon detection analysis, it had XRPD radiation diffraction data as shown in table 7.
Preparation of gentisate salt crystal form A of ninhydrin (S) -1'- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) -3-fluoropyrazin-2-yl) -1, 3-dihydrospiro [ indene-2, 4' -piperidine ] -1-amine
Weighing 50mg of (S) -1' - (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) -3-fluoropyrazin-2-yl) -1, 3-dihydrospiro [ indene-2, 4' -piperidine ] -1-amine amorphous, adding 0.25mL of ethyl acetate, stirring at 50 ℃ to obtain a clear solution, slowly adding 0.128mL of 1M gentisic acid ethanol solution into the system, stirring to separate out a large amount of solid after adding the acid, stirring at 50 ℃ for 2h, cooling, filtering and drying to obtain (S) -1' - (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) -3-fluoropyrazin-2-yl) -1, 3-dihydrospiro [ indene-2, 4' -piperidine ] -1-amine hydrogentisate form a. Upon detection analysis, it had XRPD radiation diffraction data as shown in table 8.
2. Solid stability test
2.1 purpose of experiment:
the physicochemical stability of the compound of the hydrochloride crystal form A of (S) -1'- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) -3-fluoropyrazin-2-yl) -1, 3-dihydrospiro [ indene-2, 4' -piperidine ] -1-amine under the illumination condition is examined, and a basis is provided for salt form screening and compound salt storage.
2.2 protocol:
weighing (S) -1'- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) -3-fluoropyrazin-2-yl) -1, 3-dihydrospiro [ indene-2, 4' -piperidine ] -1-amine hydrochloride crystal form A and about 2mg, inspecting for 5 days and 10 days in a lighting box (5000lx +/-500 lx), determining the content of the salt by using an HPLC (high performance liquid chromatography) and an external standard method, and calculating the change of related substances of the salt by using a chromatographic peak area normalization method.
2.3 Experimental results:
the physicochemical stability results of (S) -1'- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) -3-fluoropyrazin-2-yl) -1, 3-dihydrospiro [ indene-2, 4' -piperidin ] -1-amine hydrochloride form a are shown in table 4.3:
TABLE 4.3
Figure BDA0003005435270001371
2.4 conclusion of the experiment
(S) -1'- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) -3-fluoropyrazin-2-yl) -1, 3-dihydrospiro [ indene-2, 4' -piperidine ] -1-amine hydrochloride form a having a total gain of heteroatoms of less than 2% over 10 days under lighting conditions.
3. Hygroscopicity test
3.1 purpose of the experiment
The hygroscopicity of different salts of (S) -1'- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) -3-fluoropyrazin-2-yl) -1, 3-dihydrospiro [ indene-2, 4' -piperidine ] -1-amine under different relative humidity conditions is examined, and a basis is provided for salt type screening and storage.
3.2 Experimental protocol:
different salts of (S) -1'- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) -3-fluoropyrazin-2-yl) -1, 3-dihydrospiro [ indene-2, 4' -piperidine ] -1-amine were placed in saturated water vapor at different relative humidities to allow the compound to reach a dynamic equilibrium with the water vapor and the percentage of hygroscopic weight gain of the compound after equilibrium was calculated.
3.3 Experimental results:
3.3.1(S) -1'- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) -3-fluoropyrazin-2-yl) -1, 3-dihydrospiro [ indene-2, 4' -piperidine ] -1-amine the hygroscopicity results are as follows:
1) (S) -1'- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) -3-fluoropyrazin-2-yl) -1, 3-dihydrospiro [ indene-2, 4' -piperidine ] -1-amine hydrochloride form A gained 1.148% moisture absorption at RH 80% with slight hygroscopicity. And carrying out moisture absorption and desorption circulation for 1 time under the condition of 0-95% relative humidity, wherein the XRPD spectrogram of the (S) -1'- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) -3-fluoropyrazin-2-yl) -1, 3-dihydrospiro [ indene-2, 4' -piperidine ] -1-amine hydrochloride crystal form A is not changed, namely the crystal form is not transformed.
2) (S) -1'- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) -3-fluoropyrazin-2-yl) -1, 3-dihydrospiro [ indene-2, 4' -piperidine ] -1-amine benzenesulfonate form A had a moisture absorption weight gain of 1.631% with slight hygroscopicity at RH 80%. And the moisture absorption and desorption are circulated for 1 time under the condition of 0-95% relative humidity, and the XRPD spectrogram of the (S) -1'- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) -3-fluoropyrazin-2-yl) -1, 3-dihydrospiro [ indene-2, 4' -piperidine ] -1-amine benzene sulfonic acid crystal form A is not changed, namely the crystal form is not transformed.
4. Solubility experiments in different media
4.1 purpose of the experiment
The solubility of the (S) -1'- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) -3-fluoropyrazin-2-yl) -1, 3-dihydrospiro [ indene-2, 4' -piperidine ] -1-amine hydrochloride crystal form A in media with different pH values, water, artificial Simulated Gastric Fluid (SGF), fasted artificial simulated intestinal fluid (FaSSIF), non-fasted artificial simulated intestinal fluid (FeSSIF) and the like is compared, and a basis is provided for the ready-to-use drug evaluation of the salt.
4.2 protocol:
about 2mg of(s) -1'- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) -3-fluoropyrazin-2-yl) -1, 3-dihydrospiro [ indene-2, 4' -piperidin ] -1-amine were suspended in various media for 24 hours and the thermodynamic solubility of the compound at 37 ℃ was determined by HPLC, external standard method.
4.3 Experimental results: as shown in Table 4.4
TABLE 4.4
Figure BDA0003005435270001381
4.4 conclusion of the experiment
The (S) -1'- (6-amino-5- ((2-amino-3-chloropyridine-4-yl) thio) -3-fluoropyrazin-2-yl) -1, 3-dihydrospiro [ indene-2, 4' -piperidine ] -1-amine after salification has obviously improved solubility in an aqueous medium, combines the actual situation in a human body, the comprehensive consideration of the salification of (S) -1'- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) -3-fluoropyrazin-2-yl) -1, 3-dihydrospiro [ indene-2, 4' -piperidine ] -1-amine has certain advantages.
5. Polycrystal screening experiment
5.1 purpose of experiment:
and finding out a relatively stable crystal form of the hydrochloride through polycrystal screening.
5.2 protocol:
selecting an organic solvent with certain solubility and water, suspending (S) -1' - (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) -3-fluoropyrazin-2-yl) -1, 3-dihydrospiro [ indene-2, 4' -piperidine ] -1-amine in the solvent system, stirring and pulping for 1 week at room temperature, centrifuging, discarding the supernatant, drying the solid in vacuum at 40 ℃ (0.1 Mpa) overnight, measuring the XRPD of the solid, and reacting the solid with (S) -1' - (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) -3-fluoropyrazin-2-yl) -1, XRPD comparison of hydrochloride salt of 3-dihydrospiro [ indene-2, 4' -piperidine ] -1-amine.
5.3 Experimental results:
by pulping, changing crystallization solvent, crystallization mode and the like, only two crystal forms of hydrochloride are obtained, namely a crystal form A and a crystal form B. Comparing DSC spectra of the two crystal forms, the crystal form A can be judged to be a thermodynamically stable crystal form.

Claims (20)

1. An acid salt of a compound of formula (I),
Figure FDA0003005435260000011
wherein:
R1selected from hydrogen, halogen, C1-6Alkyl radical, C1-6Alkoxy radical, C1-6Hydroxyalkyl radical, C3-8Cycloalkyl, 3-12 membered heterocyclyl, C6-12Aryl or 5-12 membered heteroaryl, wherein said C1-6Alkyl radical, C1-6Alkoxy radical, C1-6Hydroxyalkyl radical, C3-8Cycloalkyl, 3-12 membered heterocyclyl, C6-12Aryl and 5-12 membered heteroaryl, optionally further substituted by deuterium, halogen, amino, nitro, hydroxy, cyano, oxo, C1-6Alkyl radical, C1-6Deuterated alkyl, C1-6Haloalkyl, C1-6Alkoxy radical, C1-6Hydroxyalkyl radical, C1-6Haloalkoxy, C2-6Alkenyl radical, C2-6Alkynyl, C3-8Cycloalkyl, 3-12 membered heterocyclyl, C6-12Aryl and 5-12 membered heteroaryl;
R2selected from hydrogen, amino or C1-6An alkyl group;
R3selected from hydrogen, halogen, amino, cyano, C1-6Alkyl radical, C1-6Alkoxy radical, C1-6Hydroxyalkyl radical, C3-8Cycloalkyl or 3-12 membered heterocyclyl, wherein said C is1-6Alkyl radical, C1-6Alkoxy radical, C1-6Hydroxyalkyl radical, C3-8Cycloalkyl and 3-12 membered heterocyclyl, optionally further substituted by deuterium, halogen, amino, nitro, hydroxy, cyano, oxo, C1-6Alkyl radical, C1-6Deuterated alkyl, C1-6Haloalkyl, C1-6Alkoxy radical, C1-6Hydroxyalkyl radical, C1-6Haloalkoxy, C2-6Alkenyl radical, C2-6Alkynyl, C 3-8Cycloalkyl, 3-12 membered heterocyclyl, C6-12Aryl and 5-12 membered heteroaryl;
R4selected from hydrogen, deuterium, halogen, amino, nitro, hydroxy, cyano, C1-6Alkyl radical, C1-6Deuterated alkyl, C1-6Haloalkyl, C1-6Alkoxy radical, C1-6Hydroxyalkyl radical, C1-6Haloalkoxy, C3-8Cycloalkyl, 3-12 membered heterocyclyl, C6-12Aryl or 5-12 membered heteroaryl;
R5and R6Each independently selected from hydrogen, deuterium, halogen, amino, nitro, hydroxy, cyano, C1-6Alkyl radical, C1-6Deuterated alkyl, C1-6Haloalkyl, C1-6Alkoxy radical, C1-6Haloalkoxy, C1-6Hydroxyalkyl radical, C2-6Alkenyl radical, C3-8Cycloalkyl, 3-12 membered heterocyclyl, C6-12Aryl or 5-12 membered heteroaryl, wherein said C1-6Alkyl radical, C1-6Deuterated alkyl, C1-6Haloalkyl, C1-6Alkoxy radical, C1-6Haloalkoxy, C1-6Hydroxyalkyl radical, C2-6Alkenyl radical, C3-8Cycloalkyl, 3-12 membered heterocyclyl, C6-12Aryl and 5-12 membered heteroaryl, optionally further substituted by deuterium, halogen, amino, nitro, hydroxy, cyano, oxo, C1-6Alkyl radical, C1-6Deuterated alkyl, C1-6Haloalkyl, C1-6Alkoxy radical, C1-6Hydroxyalkyl radical, C1-6Haloalkoxy, C2-6Alkenyl radical, C2-6Alkynyl, C3-8Cycloalkyl, 3-12 membered heterocyclyl, C6-12Aryl and 5-12 membered heteroaryl;
Or, R5And R6Are linked to form a C3-8Cycloalkyl, 3-12 membered heterocyclyl, C6-12Aryl or 5-12 membered heteroaryl, wherein said C3-8Cycloalkyl, 3-12 membered heterocyclyl, C6-12Aryl and 5-12 membered heteroaryl, optionally further substituted by one or more substituents selected from deuterium, halogen, amino, nitro, hydroxy, cyano, oxo, C1-6Alkyl radical, C1-6Deuterated alkyl, C1-6Haloalkyl, C1-6Alkoxy radical, C1-6Hydroxyalkyl radical, C1-6Haloalkoxy, C2-6Alkenyl radical, C2-6Alkynyl, C3-8Cycloalkyl, 3-12 membered heterocyclyl, C6-12Aryl and 5-12 membered heteroaryl;
the acid of the acid salt is an inorganic acid or an organic acid, preferably, the inorganic acid is selected from hydrochloric acid, sulfuric acid, nitric acid, hydrobromic acid, hydrofluoric acid, hydroiodic acid, or phosphoric acid; the organic acid is selected from 2, 5-dihydroxybenzoic acid, 1-hydroxy-2-naphthoic acid, acetic acid, dichloroacetic acid, trichloroacetic acid, acetoxy-hydroxamic acid, adipic acid, benzenesulfonic acid, 4-chlorobenzenesulfonic acid, benzoic acid, 4-acetamidobenzoic acid, 4-aminobenzoic acid, capric acid, caproic acid, caprylic acid, cinnamic acid, citric acid, cyclamic acid, camphorsulfonic acid, aspartic acid, camphoric acid, gluconic acid, glucuronic acid, glutamic acid, isoascorbic acid, lactic acid, malic acid, mandelic acid, pyroglutamic acid, tartaric acid, dodecylsulfuric acid, dibenzoyltartaric acid, ethane-1, 2-disulfonic acid, ethanesulfonic acid, formic acid, fumaric acid, galactaric acid, gentisic acid, glutaric acid, 2-ketoglutaric acid, glycolic acid, hippuric acid, isethionic acid, lactobionic acid, lactic acid, tartaric acid, citric acid, tartaric acid, citric acid, tartaric acid, citric acid, tartaric acid, citric acid, tartaric acid, citric acid, ascorbic acid, aspartic acid, lauric acid, camphoric acid, maleic acid, malonic acid, methanesulfonic acid, 1, 5-naphthalenedisulfonic acid, naphthalene-2-sulfonic acid, nicotinic acid, oleic acid, orotic acid, oxalic acid, palmitic acid, pamoic acid, propionic acid, salicylic acid, 4-aminosalicylic acid, sebacic acid, stearic acid, succinic acid, thiocyanic acid, undecylenic acid, trifluoroacetic acid, benzenesulfonic acid, p-toluenesulfonic acid or L-malic acid.
2. An acid salt according to claim 1 wherein R is1Selected from hydrogen, halogen, C1-6Alkyl radical, C1-6Hydroxyalkyl radical, C3-8Cycloalkyl or 3-12 membered heterocyclyl, wherein said C is1-6Alkyl radical, C1-6Hydroxyalkyl radical, C3-8Cycloalkyl and 3-12 membered heterocyclyl, optionally further substituted by deuterium, halogen, hydroxy, oxo, C1-6Alkyl and C3-8Cycloalkyl substituted with one or more substituents; r1Preferably hydrogen, halogen, C1-3Alkyl radical, C1-3Hydroxyalkyl radical, C3-6Cycloalkyl or 3-6 membered heterocyclyl, wherein said C is1-3Alkyl radical, C1-3Hydroxyalkyl radical, C3-6Cycloalkyl and 3-6 membered heterocyclyl, optionally further substituted by deuterium, halogen, hydroxy, oxo, C1-3Alkyl and C3-6Cycloalkyl substituted with one or more substituents; r1More preferably hydrogen, fluorine, bromine, methyl, hydroxyethyl,
Figure FDA0003005435260000021
3. The acid salt of claim 1 wherein R is2Selected from hydrogen, amino or C1-6An alkyl group; preferably hydrogen, amino or C1-3An alkyl group; more preferably hydrogen, amino, methyl, ethyl or propyl; further preferred is hydrogen, amino or methyl.
4. The acid salt of claim 1 wherein R is3Selected from hydrogen, halogen, amino, C1-6Alkyl radical, C3-8Cycloalkyl or 3-12 membered heterocyclyl, wherein said C is 1-6Alkyl radical, C3-8Cycloalkyl and 3-12 membered heterocyclyl, optionally further substituted with one or more substituents of deuterium, halogen, oxo; r3Preferably hydrogen, halogen, amino, C1-3Alkyl radical, C3-6Cycloalkyl or 3-6 membered heterocyclyl, wherein said C is1-3Alkyl radical, C3-6Cycloalkyl and 3-6 membered heterocyclyl, optionally further substituted with one or more substituents of deuterium, halogen, oxo; r3Further preferred are amino, chloro, cyclopropyl, azetidinyl, tetrahydropyrrolyl, morpholinyl, azetidinyl, pyrrolyl, morpholinyl, pyrrolyl, and thienyl,
Figure FDA0003005435260000031
5. The acid salt of claim 1 wherein R is4Selected from hydrogen, halogen, C1-6Alkyl or C3-8A cycloalkyl group; preferably hydrogen, halogen, C1-3Alkyl or C3-6A cycloalkyl group; further preferred is hydrogen, fluorine, chlorine, methyl or cyclopropyl.
6. The acid salt of claim 1 wherein R is5And R6Each independently selected from hydrogen and C1-6Alkyl radical, C1-6Haloalkyl, C2-6Alkenyl or C3-8Cycloalkyl, wherein said C1-6Alkyl radical, C1-6Haloalkyl, C2-6Alkenyl and C3-8Cycloalkyl optionally further substituted by deuterium, halogen, C1-6Substituted by one or more substituents in the alkyl group; preferably R5And R6Each independently selected from hydrogen and C1-3Alkyl radical, C2-4Alkenyl or C3-6Cycloalkyl, wherein said C 1-3Alkyl radical, C2-4Alkenyl and C3-6Cycloalkyl is optionally further substituted by hydrogen, deuterium, halogen, C1-3Substituted by one or more substituents in the alkyl group; more preferably R5And R6Each independently selected from hydrogen, methyl, ethyl, isopropyl, vinyl, cyclopropyl,
Figure FDA0003005435260000032
Or, R5And R6Are linked to form a C3-8Cycloalkyl, 6-12 aryl or 5-12 membered heteroaryl, wherein said C is3-8Cycloalkyl, 6-12 aryl and 5-12 membered heteroaryl, optionally further substituted by one or more substituents selected from deuterium, halogen or C1-6Substituted by one or more substituents in the alkyl group; preferably R5And R6Are linked to form a C3-6Cycloalkyl, phenyl or 5-6 membered heteroaryl, wherein said C3-6Cycloalkyl, phenyl and 5-6 membered heteroaryl, optionally further substituted by one or more substituents selected from deuterium, halogen or C1-3Substituted by one or more substituents in the alkyl group; more preferably R5And R6Are linked to form one
Figure FDA0003005435260000033
7. An acid salt according to claim 1, wherein the acid in the acid salt is selected from hydrochloric acid, phosphoric acid, acetic acid, benzoic acid, p-toluenesulfonic acid, succinic acid, tartaric acid, ethanesulfonic acid, benzenesulfonic acid, fumaric acid, isethionic acid, oxalic acid, hydrobromic acid or gentisic acid; preferably hydrochloric acid, ethanesulfonic acid, benzenesulfonic acid, fumaric acid, isethionic acid, oxalic acid, hydrobromic acid or gentisic acid; further preferred is hydrochloric acid or ethanesulfonic acid; and is
y is 1 or 2.
8. The acid salt of claim 1, wherein the compound has the structure of formula (II):
Figure FDA0003005435260000041
wherein:
ring A is selected from C3-8Cycloalkyl, 3-12 membered heterocyclyl, C6-10Aryl or 5-12 membered heteroaryl;
Raselected from hydrogen, deuterium, halogen, amino, nitro, hydroxy, cyano, C1-6Alkyl radical, C1-6Deuterated alkyl, C1-6Haloalkyl, C1-6Alkoxy radical, C1-6Hydroxyalkyl radical, C1-6Haloalkoxy, C2-6Alkenyl radical, C2-6Alkynyl, C3-8Cycloalkyl, 3-12 membered heterocyclyl, C6-12Aryl or 5-12 membered heteroaryl; and is
x is 0, 1, 2 or 3.
9. An acid salt according to claim 1 wherein the ring a is phenyl.
10. An acid salt according to claim 1, wherein the compound has the following structure:
Figure FDA0003005435260000042
Figure FDA0003005435260000051
11. an acid salt according to claim 10, characterized in that the acid salt is an acid salt of the compound (S) -1'- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) -3-fluoropyrazin-2-yl) -1, 3-dihydrospiro [ indene-2, 4' -piperidin ] -1-amine, the acid preferably being hydrochloric acid, ethanesulfonic acid, benzenesulfonic acid, fumaric acid, isethionic acid, oxalic acid, hydrobromic acid or gentisic acid;
or, as an acid salt of the compound (S) -8- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) pyrazin-2-yl) -2-cyclopropyl-8-azaspiro [4.5] dec-2-en-1-amine, the acid preferably being methanesulfonic acid, isethionic acid, fumaric acid, p-toluenesulfonic acid, ethanesulfonic acid, succinic acid, hydrobromic acid, hydrochloric acid, maleic acid, tartaric acid or sulfuric acid.
12. An acid salt according to claims 1 to 11 wherein the number of acids is 1 to 3, preferably 1, 1.5, 2, 2.5 or 3, more preferably 1 or 2.
13. An acid salt according to claims 1 to 11 wherein the number of acids is from 0.5 to 3, preferably 0.5, 1, 1.5, 2, 2.5 or 3, more preferably 0.5, 1 or 2;
wherein in the acid salt of (S) -1'- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) -3-fluoropyrazin-2-yl) -1, 3-dihydrospiro [ indene-2, 4' -piperidine ] -1-amine the number of acids is from 1 to 3, preferably 1, 1.5, 2, 2.5 or 3, more preferably 1 or 2; or the like, or, alternatively,
in an acid salt of (S) -8- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) pyrazin-2-yl) -2-cyclopropyl-8-azaspiro [4.5] dec-2-en-1-amine, the number of acids is 0.2 to 3; preferably 0.2, 0.5, 1, 1.5, 2, 2.5 or 3; more preferably 0.5, 1, 2 or 3; further preferably 0.5 or 1.
14. An acid salt according to claim 11, characterized in that the acid salt of the compound (S) -1'- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) -3-fluoropyrazin-2-yl) -1, 3-dihydrospiro [ indene-2, 4' -piperidin ] -1-amine is in crystalline form, preferably in crystalline form as hydrochloride, ethanesulfonate, benzenesulfonate, fumarate, isethionate, oxalate, hydrobromide or gentisate;
Wherein the crystalline form of the hydrochloride salt comprises form a and form B, form a having an X-ray powder diffraction pattern with diffraction peaks at 15.1 °, 21.0 °, 22.6 ° and 23.7 ° 2 Θ, preferably further comprising diffraction peaks at 8.4 °, 9.4 °, 16.7 °, 18.4 ° and 25.5 ° 2 Θ, more preferably further comprising diffraction peaks at 14.5 °, 19.2 °, 20.5 °, 24.8 °, 25.1 °, 27.8 ° and 31.4 ° 2 Θ, further preferably further comprising diffraction peaks at 11.3 °, 16.3 °, 23.4 °, 24.1 °, 26.4 °, 28.2 °, 29.0 °, 30.6 °, 32.3 °, 32.8 °, 35.6 ° and 37.0 ° 2 Θ, most preferably having an XRPD pattern substantially as shown in figure 1, or a pattern substantially as shown in figure 2 DSC, or a pattern substantially as shown in figure 3 TGA;
said crystalline form B having an X-ray powder diffraction pattern with diffraction peaks at 6.9 °, 13.8 °, 18.0 ° and 21.7 ° 2 Θ, preferably further comprising diffraction peaks at 8.8 °, 9.1 °, 13.6 ° and 23.9 ° 2 Θ, more preferably further comprising diffraction peaks at 18.5 °, 25.0 °, 26.2 ° and 27.5 ° 2 Θ, further preferably having a DSC pattern substantially as shown in figure 4, or a TGA pattern substantially as shown in figure 5;
The ethanesulfonate comprises form a having an X-ray powder diffraction pattern with diffraction peaks at 8.3 °, 11.6 °, and 18.3 ° 2 Θ, preferably further comprising diffraction peaks at 13.4 °, 15.3 °, 18.5 °, 20.4 °, 21.0 °, and 23.4 ° 2 Θ, more preferably further comprising diffraction peaks at 10.7 °, 12.6 °, 14.1 °, 21.9 °, 22.5 °, and 25.3 ° 2 Θ, further preferably having an XRPD pattern substantially as shown in figure 6, or a DSC pattern substantially as shown in figure 7, or a TGA pattern substantially as shown in figure 8;
besylate includes form a having an X-ray powder diffraction pattern having diffraction peaks at 12.8 °, 15.3 °, 21.4 ° and 25.8 ° 2 Θ, preferably further comprising diffraction peaks at 7.4 °, 11.1 ° and 22.3 ° 2 θ, more preferably further comprising diffraction peaks at 14.8 °, 16.2 °, 18.1 °, 19.8 °, 20.9 °, 23.8 ° and 24.7 ° 2 θ, further preferably further comprising diffraction peaks at 15.9 °, 16.5 °, 19.0 °, 22.8 °, 27.3 ° and 28.3 ° 2 θ, or a DSC substantially as shown in figure 9, or a TGA substantially as shown in figure 10;
the fumarate salt comprises form a having an X-ray powder diffraction pattern with diffraction peaks at 8.6 ° and 16.9 ° 2 Θ, preferably further comprising diffraction peaks at 11.5 °, 13.0 ° and 22.1 ° 2 θ, more preferably further comprising diffraction peaks at 14.1 °, 18.6 °, 23.2 °, 23.9 ° and 26.8 ° 2 θ, further preferably further comprising diffraction peaks at 19.5 °, 23.5 ° and 26.0 ° 2 θ, or a DSC pattern substantially as shown in figure 11, or a TGA pattern substantially as shown in figure 12;
Isethionate comprises form a having an X-ray powder diffraction pattern with diffraction peaks at 12.3 °, 16.5 ° and 24.8 ° 2 Θ, preferably further comprising diffraction peaks at 15.0 °, 18.8 °, 26.1 ° and 27.5 ° 2 Θ, more preferably further comprising diffraction peaks at 14.0 °, 16.2 ° and 18.7 ° 2 Θ, or having a DSC pattern substantially as shown in figure 13, or having a TGA pattern substantially as shown in figure 14;
the oxalate salt comprises a form A, wherein the X-ray powder diffraction pattern of the form A has diffraction peaks at 14.3 degrees, 17.8 degrees and 23.3 degrees of 2 theta, preferably also comprises diffraction peaks at 6.6 degrees, 11.9 degrees, 20.2 degrees and 26.1 degrees of 2 theta, more preferably further comprises diffraction peaks at 8.7 degrees, 15.7 degrees, 24.5 degrees, 27.6 degrees and 28.9 degrees of 2 theta, and further preferably has a DSC pattern which is basically as shown in figure 15 and a TGA pattern which is basically as shown in figure 16;
the hydrobromide salt comprises form a having an X-ray powder diffraction pattern with diffraction peaks at 13.0 °, 19.3 ° and 25.2 ° 2 θ, preferably further comprising diffraction peaks at 7.1 °, 24.3 °, 26.6 ° and 27.1 ° 2 θ, more preferably further comprising diffraction peaks at 6.0 °, 12.7 °, 14.0 °, 19.9 °, 21.6 °, 22.0 °, 25.7 ° and 26.3 ° 2 θ; still further preferably, further comprising having diffraction peaks at 11.1 °, 12.0 °, 20.3 °, 22.5 °, 27.9 ° and 30.5 ° 2 θ;
The gentisate salt comprises a form a having an X-ray powder diffraction pattern with diffraction peaks at 12.4 °, 17.6 °, 20.8 ° and 22.2 ° 2 Θ, preferably further comprising diffraction peaks at 7.5 °, 8.5 °, 15.8 ° and 25.8 ° 2 θ, more preferably further comprising diffraction peaks at 5.7 °, 10.6 °, 14.8 °, 17.1 °, 18.9 ° and 24.9 ° 2 θ.
15. An acid salt according to claim 11, characterized in that the acid salt of (S) -8- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) pyrazin-2-yl) -2-cyclopropyl-8-azaspiro [4.5] dec-2-en-1-amine is in crystalline form, preferably in the form of methanesulfonate, isethionate, fumarate, p-toluenesulfonate, ethanesulfonate, succinate, hydrobromide, hydrochloride, maleate, tartrate or sulfate;
wherein the content of the first and second substances,
the mesylate crystal form is mesylate crystal form A, the X-ray powder diffraction pattern of the mesylate crystal form A has a diffraction peak at 20.0 +/-0.2 degrees, or a diffraction peak at 17.8 + -0.2 deg., or a diffraction peak at 24.2 + -0.2 deg., or a diffraction peak at 22.2 + -0.2 deg., or a diffraction peak at 24.6 + -0.2 deg., or a diffraction peak at 12.5 + -0.2 deg., or a diffraction peak at 28.6 + -0.2 deg., or a diffraction peak at 18.8 + -0.2 deg., or a diffraction peak at 11.0 + -0.2 deg., or a diffraction peak at 14.2 + -0.2 deg., preferably including any 2-5 of the above diffraction peaks, or 3-5, or 3-6, or 3-8, or 5-8, or 6-8, more preferably including any 6, 7, or 8 thereof,
Preferably, the X-ray powder diffraction pattern of mesylate form A comprises at least one or more diffraction peaks, preferably 2 thereof, more preferably 3 thereof, and optionally further comprises at least one of 22.2 + -0.2 °, 24.6 + -0.2 °, 12.5 + -0.2 °, 28.6 + -0.2 °, 18.8 + -0.2 °, preferably 2, 3, 4 or 5 thereof, at 2 theta, 20.0 + -0.2 °, 17.8 + -0.2 °, 24.2 + -0.2 °, and 24.2 + -0.2 °,
for example, the mesylate salt form a has an X-ray powder diffraction pattern with diffraction peaks at the following positions 2 θ:
at 20.0 +/-0.2 degrees and 17.8 +/-0.2 degrees,
or 17.8 +/-0.2 degrees and 24.2 +/-0.2 degrees,
or 20.0 +/-0.2 degrees and 24.2 +/-0.2 degrees,
or 20.0 +/-0.2 degrees, 17.8 +/-0.2 degrees and 24.2 +/-0.2 degrees,
or 17.8 +/-0.2 degrees, 24.2 +/-0.2 degrees and 22.2 +/-0.2 degrees,
or 20.0 +/-0.2 degrees, 17.8 +/-0.2 degrees, 24.2 +/-0.2 degrees and 22.2 +/-0.2 degrees,
or 17.8 +/-0.2 degrees, 24.2 +/-0.2 degrees, 22.2 +/-0.2 degrees and 24.6 +/-0.2 degrees,
or 20.0 +/-0.2 degrees, 17.8 +/-0.2 degrees, 24.2 +/-0.2 degrees, 22.2 +/-0.2 degrees and 24.6 +/-0.2 degrees,
or 17.8 +/-0.2 degrees, 24.2 +/-0.2 degrees, 22.2 +/-0.2 degrees, 24.6 +/-0.2 degrees and 12.5 +/-0.2 degrees,
more preferably, the X-ray powder diffraction pattern of mesylate form A optionally further comprises one or more diffraction peaks, preferably at least any 2-3, or 4-5, or 6-7, more preferably, any 2, 3, 4, 5, 6, 7, thereof at 2 θ of 11.0 ± 0.2 °, 14.2 ± 0.2 °, 28.2 ± 0.2 °, 21.9 ± 0.2 °, 26.6 ± 0.2 °, 21.2 ± 0.2 °, 29.7 ± 0.2 °, more preferably, at least any 2, 3, 4, 5, 6, 7,
For example, the mesylate salt form a has an X-ray powder diffraction pattern with diffraction peaks at the following positions 2 θ:
11.0 +/-0.2 degrees, 14.2 +/-0.2 degrees, 28.2 +/-0.2 degrees, 21.9 +/-0.2 degrees, 26.6 +/-0.2 degrees and 21.2 +/-0.2 degrees,
or 14.2 +/-0.2 degrees, 28.2 +/-0.2 degrees, 21.9 +/-0.2 degrees, 26.6 +/-0.2 degrees, 21.2 +/-0.2 degrees and 29.7 +/-0.2 degrees,
or 11.0 +/-0.2 degrees, 14.2 +/-0.2 degrees, 21.9 +/-0.2 degrees, 26.6 +/-0.2 degrees, 21.2 +/-0.2 degrees and 29.7 +/-0.2 degrees,
further preferably, the mesylate form A has an X-ray powder diffraction pattern comprising diffraction peaks at one or more of 20.0 + -0.2 °, 17.8 + -0.2 °, 24.2 + -0.2 °, 22.2 + -0.2 °, 24.6 + -0.2 °, 12.5 + -0.2 °, 28.6 + -0.2 °, 18.8 + -0.2 °, 11.0 + -0.2 °, 14.2 + -0.2 °, 28.2 + -0.2 °, 21.9 + -0.2 °, 26.6 + -0.2 °, 21.2 + -0.2 °, 29.7 + -0.2 ° 2 θ, preferably comprising diffraction peaks at any of 4, 5, 6, 8, 10,
for example, the mesylate salt form a has an X-ray powder diffraction pattern with diffraction peaks at the following positions 2 θ:
at 20.0 +/-0.2 degrees, 17.8 +/-0.2 degrees, 24.2 +/-0.2 degrees and 29.7 +/-0.2 degrees,
or 17.8 +/-0.2 degrees, 24.2 +/-0.2 degrees, 22.2 +/-0.2 degrees and 21.2 +/-0.2 degrees,
or 24.2 +/-0.2 degrees, 22.2 +/-0.2 degrees, 24.6 +/-0.2 degrees, 12.5 +/-0.2 degrees and 28.6 +/-0.2 degrees,
Or 22.2 +/-0.2 degrees, 24.6 +/-0.2 degrees, 12.5 +/-0.2 degrees, 28.6 +/-0.2 degrees and 18.8 +/-0.2 degrees,
or 20.0 +/-0.2 degrees, 17.8 +/-0.2 degrees, 24.2 +/-0.2 degrees, 24.6 +/-0.2 degrees, 12.5 +/-0.2 degrees and 28.6 +/-0.2 degrees,
or 17.8 +/-0.2 degrees, 24.2 +/-0.2 degrees, 22.2 +/-0.2 degrees, 24.6 +/-0.2 degrees, 28.6 +/-0.2 degrees and 18.8 +/-0.2 degrees,
or 24.2 +/-0.2 degrees, 22.2 +/-0.2 degrees, 24.6 +/-0.2 degrees, 12.5 +/-0.2 degrees, 28.6 +/-0.2 degrees, 18.8 +/-0.2 degrees, 11.0 +/-0.2 degrees and 14.2 +/-0.2 degrees,
or 22.2 +/-0.2 degrees, 24.6 +/-0.2 degrees, 12.5 +/-0.2 degrees, 28.6 +/-0.2 degrees, 18.8 +/-0.2 degrees, 11.0 +/-0.2 degrees, 14.2 +/-0.2 degrees and 28.2 +/-0.2 degrees,
or 20.0 +/-0.2 degrees, 17.8 +/-0.2 degrees, 24.2 +/-0.2 degrees, 22.2 +/-0.2 degrees, 24.6 +/-0.2 degrees, 12.5 +/-0.2 degrees, 28.6 +/-0.2 degrees, 18.8 +/-0.2 degrees, 11.0 +/-0.2 degrees and 14.2 +/-0.2 degrees,
or 17.8 +/-0.2 degrees, 24.2 +/-0.2 degrees, 22.2 +/-0.2 degrees, 24.6 +/-0.2 degrees, 12.5 +/-0.2 degrees, 28.6 +/-0.2 degrees, 18.8 +/-0.2 degrees, 11.0 +/-0.2 degrees, 14.2 +/-0.2 degrees and 28.2 +/-0.2 degrees,
still more preferably, the mesylate form a has an X-ray powder diffraction pattern, as shown in figure 25, or a DSC pattern, as shown in figure 26;
the isethionate salt form A is isethionate salt form A, form A having an X-ray powder diffraction pattern with a diffraction peak at 20.1 + -0.2 °, or a diffraction peak at 18.7 + -0.2 °, or a diffraction peak at 23.4 + -0.2 °, or a diffraction peak at 18.1 + -0.2 °, or a diffraction peak at 20.9 + -0.2 °, or a diffraction peak at 23.2 + -0.2 °, or a diffraction peak at 12.6 + -0.2 °, or a diffraction peak at 13.3 + -0.2 °, or a diffraction peak at 11.6 + -0.2 °, or a diffraction peak at 10.4 + -0.2 °, preferably comprising any of the above diffraction peaks 2 to 5, or 3 to 6, or 3 to 8, or 5 to 8, or 6 to 8, more preferably comprising any of the above diffraction peaks, 7 or 8,
Preferably, the isethionate salt form a has an X-ray powder diffraction pattern comprising at least one diffraction peak located at one or more of 20.1 ± 0.2 °, 18.7 ± 0.2 °, 23.4 ± 0.2 °, preferably comprising 2, more preferably 3, and optionally further comprising at least one of 18.1 ± 0.2 °, 20.9 ± 0.2 °, 23.2 ± 0.2 °, 12.6 ± 0.2 °, 13.3 ± 0.2 °, preferably comprising 2, 3, 4 or 5, of 2 Θ,
for example, the isethionate salt form a has an X-ray powder diffraction pattern with diffraction peaks at the following positions 2 θ:
at 20.1 +/-0.2 degrees and 18.7 +/-0.2 degrees,
or 18.7 +/-0.2 degrees and 23.4 +/-0.2 degrees,
or 20.1 +/-0.2 degrees, 18.7 +/-0.2 degrees and 23.4 +/-0.2 degrees,
or at 18.7 +/-0.2 degrees, 23.4 +/-0.2 degrees and 18.1 +/-0.2 degrees,
or 20.1 +/-0.2 degrees, 18.7 +/-0.2 degrees, 23.4 +/-0.2 degrees and 18.1 +/-0.2 degrees,
or at 18.7 +/-0.2 degrees, 23.4 +/-0.2 degrees, 18.1 +/-0.2 degrees and 20.9 +/-0.2 degrees,
or 20.1 +/-0.2 degrees, 18.7 +/-0.2 degrees, 23.4 +/-0.2 degrees, 18.1 +/-0.2 degrees and 20.9 +/-0.2 degrees,
or at 18.7 + -0.2 deg., 23.4 + -0.2 deg., 18.1 + -0.2 deg., 20.9 + -0.2 deg. and 23.2 + -0.2 deg.,
more preferably, the isethionate salt form A optionally further comprises one or more diffraction peaks at 11.6 + -0.2 deg., 10.4 + -0.2 deg., 16.8 + -0.2 deg., 19.8 + -0.2 deg., 24.5 + -0.2 deg., 28.0 + -0.2 deg., 24.9 + -0.2 deg. 2 θ, preferably at least any 2-3, or 4-5, or 6-7, more preferably any 2, 3, 4, 5, 6, 7,
For example, the isethionate salt form a has an X-ray powder diffraction pattern with diffraction peaks at the following positions 2 θ:
11.6 +/-0.2 degrees, 10.4 +/-0.2 degrees, 16.8 +/-0.2 degrees, 19.8 +/-0.2 degrees, 24.5 +/-0.2 degrees and 28.0 +/-0.2 degrees,
or 10.4 +/-0.2 degrees, 16.8 +/-0.2 degrees, 19.8 +/-0.2 degrees, 24.5 +/-0.2 degrees, 28.0 +/-0.2 degrees and 24.9 +/-0.2 degrees,
or 11.6 +/-0.2 degrees, 10.4 +/-0.2 degrees, 19.8 +/-0.2 degrees, 24.5 +/-0.2 degrees, 28.0 +/-0.2 degrees and 24.9 +/-0.2 degrees,
further preferably, the isethionate salt form a has an X-ray powder diffraction pattern comprising diffraction peaks at one or more of 20.1 ± 0.2 °, 18.7 ± 0.2 °, 23.4 ± 0.2 °, 18.1 ± 0.2 °, 20.9 ± 0.2 °, 23.2 ± 0.2 °, 12.6 ± 0.2 °, 13.3 ± 0.2 °, 11.6 ± 0.2 °, 10.4 ± 0.2 °, 16.8 ± 0.2 °, 19.8 ± 0.2 °, 24.5 ± 0.2 °, 28.0 ± 0.2 °, and 24.9 ± 0.2 ° 2 Θ; preferably, the compound contains diffraction peaks at 4, 5, 6, 8 and 10,
for example, the isethionate salt form a has an X-ray powder diffraction pattern with diffraction peaks at the following positions 2 θ:
20.1 +/-0.2 degrees, 18.1 +/-0.2 degrees, 20.9 +/-0.2 degrees and 23.2 +/-0.2 degrees,
or 18.7 +/-0.2 degrees, 18.1 +/-0.2 degrees, 20.9 +/-0.2 degrees and 23.2 +/-0.2 degrees,
Or 23.4 +/-0.2 degrees, 18.1 +/-0.2 degrees, 20.9 +/-0.2 degrees, 23.2 +/-0.2 degrees and 12.6 +/-0.2 degrees,
or 18.1 +/-0.2 degrees, 20.9 +/-0.2 degrees, 23.2 +/-0.2 degrees, 12.6 +/-0.2 degrees and 13.3 +/-0.2 degrees,
or 20.1 +/-0.2 degrees, 18.7 +/-0.2 degrees, 23.4 +/-0.2 degrees, 18.1 +/-0.2 degrees, 20.9 +/-0.2 degrees and 23.2 +/-0.2 degrees,
or 18.7 +/-0.2 degrees, 23.4 +/-0.2 degrees, 18.1 +/-0.2 degrees, 20.9 +/-0.2 degrees, 23.2 +/-0.2 degrees and 12.6 +/-0.2 degrees,
or 23.4 +/-0.2 degrees, 18.1 +/-0.2 degrees, 20.9 +/-0.2 degrees, 23.2 +/-0.2 degrees, 12.6 +/-0.2 degrees, 13.3 +/-0.2 degrees, 11.6 +/-0.2 degrees and 10.4 +/-0.2 degrees,
or 18.1 +/-0.2 degrees, 20.9 +/-0.2 degrees, 23.2 +/-0.2 degrees, 12.6 +/-0.2 degrees, 13.3 +/-0.2 degrees, 11.6 +/-0.2 degrees, 10.4 +/-0.2 degrees and 16.8 +/-0.2 degrees,
or 20.1 +/-0.2 degrees, 18.7 +/-0.2 degrees, 23.4 +/-0.2 degrees, 18.1 +/-0.2 degrees, 20.9 +/-0.2 degrees, 23.2 +/-0.2 degrees, 12.6 +/-0.2 degrees, 13.3 +/-0.2 degrees, 11.6 +/-0.2 degrees and 10.4 +/-0.2 degrees,
or 18.7 +/-0.2 degrees, 23.4 +/-0.2 degrees, 18.1 +/-0.2 degrees, 20.9 +/-0.2 degrees, 23.2 +/-0.2 degrees, 12.6 +/-0.2 degrees, 13.3 +/-0.2 degrees, 11.6 +/-0.2 degrees, 10.4 +/-0.2 degrees and 16.8 +/-0.2 degrees,
still more preferably, isethionate salt form a has an X-ray powder diffraction pattern as shown in figure 27, or a DSC pattern as shown in figure 28;
The fumarate crystal form is fumarate crystal form A, the X-ray powder diffraction pattern of the fumarate crystal form A has diffraction peaks at 18.7 +/-0.2 degrees, or a diffraction peak at 23.1 + -0.2 deg., or a diffraction peak at 20.1 + -0.2 deg., or a diffraction peak at 11.3 + -0.2 deg., or a diffraction peak at 25.0 + -0.2 deg., or a diffraction peak at 12.6 + -0.2 deg., or a diffraction peak at 22.0 + -0.2 deg., or a diffraction peak at 18.2 + -0.2 deg., or a diffraction peak at 27.8 + -0.2 deg., or a diffraction peak at 14.6 + -0.2 deg., preferably any 2-5 of the above diffraction peaks, or 3-5, or 3-6, or 3-8, or 5-8, or 6-8, more preferably including any 6, 7, or 8 thereof,
preferably, the fumarate salt form a has an X-ray powder diffraction pattern comprising at least one diffraction peak, preferably 2, more preferably 3, at least one diffraction peak, preferably 2, 3, 4 or 5, at 2 θ of 18.7 ± 0.2 °, 23.1 ± 0.2 °, 20.1 ± 0.2 °, optionally further comprising at least one diffraction peak, preferably 2, 3, 4 or 5, at 2 θ of 11.3 ± 0.2 °, 25.0 ± 0.2 °, 12.6 ± 0.2 °, 22.0 ± 0.2 °, 18.2 ± 0.2 °,
For example, the fumarate salt form a has an X-ray powder diffraction pattern with diffraction peaks at the following positions 2 θ:
at 18.7 +/-0.2 degrees and 23.1 +/-0.2 degrees,
or at 23.1 + -0.2 deg. and 20.1 + -0.2 deg.,
or at 18.7 +/-0.2 degrees, 23.1 +/-0.2 degrees and 20.1 +/-0.2 degrees,
or at 23.1 +/-0.2 degrees, 20.1 +/-0.2 degrees and 11.3 +/-0.2 degrees,
or at 18.7 + -0.2 deg., 23.1 + -0.2 deg., 20.1 + -0.2 deg. and 11.3 + -0.2 deg.,
or at 23.1 + -0.2 deg., 20.1 + -0.2 deg., 11.3 + -0.2 deg. and 25.0 + -0.2 deg.,
or at 18.7 + -0.2 deg., 23.1 + -0.2 deg., 20.1 + -0.2 deg., 11.3 + -0.2 deg. and 25.0 + -0.2 deg.,
or at 23.1 + -0.2 deg., 20.1 + -0.2 deg., 11.3 + -0.2 deg., 25.0 + -0.2 deg. and 12.6 + -0.2 deg.,
more preferably, the X-ray powder diffraction pattern of form A of the fumarate salt optionally further comprises one or more diffraction peaks, preferably at least any 2-3, or 4-5, or 6-7 diffraction peaks, at any 2, 3, 4, 5, 6, 7 diffraction peaks, at 2 θ of 27.8 ± 0.2 °, 14.6 ± 0.2 °, 5.1 ± 0.2 °, 15.4 ± 0.2 °, 28.8 ± 0.2 °, 13.9 ± 0.2 °, 20.5 ± 0.2 °, more preferably at least any 2-3, or 4, 5, 6, 7,
for example, the fumarate salt form a has an X-ray powder diffraction pattern with diffraction peaks at the following positions 2 θ:
27.8 +/-0.2 degrees, 14.6 +/-0.2 degrees, 5.1 +/-0.2 degrees, 15.4 +/-0.2 degrees, 28.8 +/-0.2 degrees and 13.9 +/-0.2 degrees,
or 14.6 +/-0.2 degrees, 5.1 +/-0.2 degrees, 15.4 +/-0.2 degrees, 28.8 +/-0.2 degrees, 13.9 +/-0.2 degrees and 20.5 +/-0.2 degrees,
or 27.8 +/-0.2 degrees, 14.6 +/-0.2 degrees, 15.4 +/-0.2 degrees, 28.8 +/-0.2 degrees, 13.9 +/-0.2 degrees and 20.5 +/-0.2 degrees,
further preferably, the fumarate salt form a has an X-ray powder diffraction pattern comprising diffraction peaks at one or more of 18.7 ± 0.2 °, 23.1 ± 0.2 °, 20.1 ± 0.2 °, 11.3 ± 0.2 °, 25.0 ± 0.2 °, 12.6 ± 0.2 °, 22.0 ± 0.2 °, 18.2 ± 0.2 °, 27.8 ± 0.2 °, 14.6 ± 0.2 °, 5.1 ± 0.2 °, 15.4 ± 0.2 °, 28.8 ± 0.2 °, 13.9 ± 0.2 °, 20.5 ± 0.2 ° 2 Θ, preferably comprising diffraction peaks at 4, 5, 6, 8, 10, optionally,
for example, the fumarate salt form a has an X-ray powder diffraction pattern with diffraction peaks at the following positions 2 θ:
at 18.7 +/-0.2 degrees, 20.1 +/-0.2 degrees, 11.3 +/-0.2 degrees and 25.0 +/-0.2 degrees,
or at 23.1 +/-0.2 degrees, 11.3 +/-0.2 degrees, 25.0 +/-0.2 degrees and 12.6 +/-0.2 degrees,
or 20.1 +/-0.2 degrees, 11.3 +/-0.2 degrees, 25.0 +/-0.2 degrees, 12.6 +/-0.2 degrees and 22.0 +/-0.2 degrees,
or 11.3 +/-0.2 degrees, 25.0 +/-0.2 degrees, 12.6 +/-0.2 degrees, 22.0 +/-0.2 degrees and 18.2 +/-0.2 degrees,
Or 18.7 +/-0.2 degrees, 23.1 +/-0.2 degrees, 20.1 +/-0.2 degrees, 11.3 +/-0.2 degrees, 25.0 +/-0.2 degrees and 12.6 +/-0.2 degrees,
or 23.1 +/-0.2 degrees, 20.1 +/-0.2 degrees, 11.3 +/-0.2 degrees, 25.0 +/-0.2 degrees, 12.6 +/-0.2 degrees and 22.0 +/-0.2 degrees,
or 20.1 +/-0.2 degrees, 11.3 +/-0.2 degrees, 25.0 +/-0.2 degrees, 12.6 +/-0.2 degrees, 22.0 +/-0.2 degrees, 18.2 +/-0.2 degrees, 27.8 +/-0.2 degrees and 14.6 +/-0.2 degrees,
or 11.3 +/-0.2 degrees, 25.0 +/-0.2 degrees, 12.6 +/-0.2 degrees, 22.0 +/-0.2 degrees, 18.2 +/-0.2 degrees, 27.8 +/-0.2 degrees, 14.6 +/-0.2 degrees and 5.1 +/-0.2 degrees,
or 18.7 +/-0.2 degrees, 23.1 +/-0.2 degrees, 20.1 +/-0.2 degrees, 11.3 +/-0.2 degrees, 25.0 +/-0.2 degrees, 12.6 +/-0.2 degrees, 22.0 +/-0.2 degrees, 18.2 +/-0.2 degrees, 27.8 +/-0.2 degrees and 14.6 +/-0.2 degrees,
or 23.1 +/-0.2 degrees, 20.1 +/-0.2 degrees, 11.3 +/-0.2 degrees, 25.0 +/-0.2 degrees, 12.6 +/-0.2 degrees, 22.0 +/-0.2 degrees, 18.2 +/-0.2 degrees, 27.8 +/-0.2 degrees, 14.6 +/-0.2 degrees and 5.1 +/-0.2 degrees,
still further preferably, fumarate salt form a has an X-ray powder diffraction pattern as shown in figure 29, or a DSC pattern as shown in figure 30;
the crystalline form of p-toluenesulfonate is crystalline form A having an X-ray powder diffraction pattern with a diffraction peak at 23.2 + -0.2 deg., or a diffraction peak at 11.2 + -0.2 deg., or a diffraction peak at 19.4 + -0.2 deg., or a diffraction peak at 16.6 + -0.2 deg., or a diffraction peak at 21.1 + -0.2 deg., or a diffraction peak at 17.3 + -0.2 deg., or a diffraction peak at 16.8 + -0.2 deg., or a diffraction peak at 4.3 + -0.2 deg., or a diffraction peak at 18.2 + -0.2 deg., or a diffraction peak at 22.0 + -0.2 deg., preferably comprising any of the above diffraction peaks 2-5, or 3-6, or 3-8, or 5-8, or 6-8, more preferably comprising any of the above diffraction peaks, 7-8 or 7-8,
Preferably, the crystalline form a of p-toluenesulfonate has an X-ray powder diffraction pattern comprising at least one or more diffraction peaks, preferably 2, more preferably 3, and optionally further comprising at least one of 16.6 ± 0.2 °, 21.1 ± 0.2 °, 17.3 ± 0.2 °, 16.8 ± 0.2 °, 4.3 ± 0.2 °, preferably 2, 3, 4 or 5, 2 Θ,
for example, the p-toluenesulfonate form a has an X-ray powder diffraction pattern with diffraction peaks at the following positions 2 θ:
at 23.2 +/-0.2 degrees and 11.2 +/-0.2 degrees,
or 11.2 +/-0.2 degrees and 19.4 +/-0.2 degrees,
or at 23.2 + -0.2 deg., 11.2 + -0.2 deg. and 19.4 + -0.2 deg.,
or 11.2 +/-0.2 degrees, 19.4 +/-0.2 degrees and 16.6 +/-0.2 degrees,
or at 23.2 + -0.2 deg., 11.2 + -0.2 deg., 19.4 + -0.2 deg. and 16.6 + -0.2 deg.,
or 11.2 +/-0.2 degrees, 19.4 +/-0.2 degrees, 16.6 +/-0.2 degrees and 21.1 +/-0.2 degrees,
or at 23.2 + -0.2 deg., 11.2 + -0.2 deg., 19.4 + -0.2 deg., 16.6 + -0.2 deg. and 21.1 + -0.2 deg.,
or 11.2 +/-0.2 degrees, 19.4 +/-0.2 degrees, 16.6 +/-0.2 degrees, 21.1 +/-0.2 degrees and 17.3 +/-0.2 degrees,
more preferably, the X-ray powder diffraction pattern of crystalline form A of the tosylate salt optionally further comprises one or more diffraction peaks at 2 θ of 18.2. + -. 0.2 °, 22.0. + -. 0.2 °, 29.2. + -. 0.2 °, 20.7. + -. 0.2 °, 9.6. + -. 0.2 °, 26.9. + -. 0.2 °, 13.2. + -. 0.2 °, preferably at least any 2-3, or 4-5, or 6-7 thereof, further preferably any 2, 3, 4, 5, 6, 7 thereof,
For example, the p-toluenesulfonate form a has an X-ray powder diffraction pattern with diffraction peaks at the following positions 2 θ:
18.2 +/-0.2 degrees, 22.0 +/-0.2 degrees, 29.2 +/-0.2 degrees, 20.7 +/-0.2 degrees, 9.6 +/-0.2 degrees and 26.9 +/-0.2 degrees,
or 22.0 +/-0.2 degrees, 29.2 +/-0.2 degrees, 20.7 +/-0.2 degrees, 9.6 +/-0.2 degrees, 26.9 +/-0.2 degrees and 13.2 +/-0.2 degrees,
or 18.2 +/-0.2 degrees, 22.0 +/-0.2 degrees, 20.7 +/-0.2 degrees, 9.6 +/-0.2 degrees, 26.9 +/-0.2 degrees and 13.2 +/-0.2 degrees,
further preferably, the crystalline form a of tosylate has an X-ray powder diffraction pattern comprising diffraction peaks at one or more of 23.2 ± 0.2 °, 11.2 ± 0.2 °, 19.4 ± 0.2 °, 16.6 ± 0.2 °, 21.1 ± 0.2 °, 17.3 ± 0.2 °, 16.8 ± 0.2 °, 4.3 ± 0.2 °, 18.2 ± 0.2 °, 22.0 ± 0.2 °, 29.2 ± 0.2 °, 20.7 ± 0.2 °, 9.6 ± 0.2 °, 26.9 ± 0.2 °, 13.2 ± 0.2 ° 2 Θ, preferably comprising diffraction peaks at any of 4, 5, 6, 8, 10,
for example, the p-toluenesulfonate form a has an X-ray powder diffraction pattern with diffraction peaks at the following positions 2 θ:
23.2 +/-0.2 degrees, 19.4 +/-0.2 degrees, 16.6 +/-0.2 degrees and 21.1 +/-0.2 degrees,
or 11.2 +/-0.2 degrees, 16.6 +/-0.2 degrees, 21.1 +/-0.2 degrees and 17.3 +/-0.2 degrees,
or 19.4 +/-0.2 degrees, 16.6 +/-0.2 degrees, 21.1 +/-0.2 degrees, 17.3 +/-0.2 degrees and 16.8 +/-0.2 degrees,
Or 16.6 +/-0.2 degrees, 21.1 +/-0.2 degrees, 17.3 +/-0.2 degrees, 16.8 +/-0.2 degrees and 4.3 +/-0.2 degrees,
or 23.2 +/-0.2 degrees, 11.2 +/-0.2 degrees, 19.4 +/-0.2 degrees, 16.6 +/-0.2 degrees, 21.1 +/-0.2 degrees and 17.3 +/-0.2 degrees,
or 11.2 +/-0.2 degrees, 19.4 +/-0.2 degrees, 16.6 +/-0.2 degrees, 21.1 +/-0.2 degrees, 17.3 +/-0.2 degrees and 16.8 +/-0.2 degrees,
or 19.4 +/-0.2 degrees, 16.6 +/-0.2 degrees, 21.1 +/-0.2 degrees, 17.3 +/-0.2 degrees, 16.8 +/-0.2 degrees, 4.3 +/-0.2 degrees, 18.2 +/-0.2 degrees and 22.0 +/-0.2 degrees,
or 16.6 +/-0.2 degrees, 21.1 +/-0.2 degrees, 17.3 +/-0.2 degrees, 16.8 +/-0.2 degrees, 4.3 +/-0.2 degrees, 18.2 +/-0.2 degrees, 22.0 +/-0.2 degrees and 29.2 +/-0.2 degrees,
or 23.2 +/-0.2 degrees, 11.2 +/-0.2 degrees, 19.4 +/-0.2 degrees, 16.6 +/-0.2 degrees, 21.1 +/-0.2 degrees, 17.3 +/-0.2 degrees, 16.8 +/-0.2 degrees, 4.3 +/-0.2 degrees, 18.2 +/-0.2 degrees and 22.0 +/-0.2 degrees,
or 11.2 +/-0.2 degrees, 19.4 +/-0.2 degrees, 16.6 +/-0.2 degrees, 21.1 +/-0.2 degrees, 17.3 +/-0.2 degrees, 16.8 +/-0.2 degrees, 4.3 +/-0.2 degrees, 18.2 +/-0.2 degrees, 22.0 +/-0.2 degrees and 29.2 +/-0.2 degrees,
still further preferably, form a of the p-toluenesulfonate salt has an X-ray powder diffraction pattern as shown in fig. 31 or a DSC pattern as shown in fig. 32;
the ethanesulfonate crystal form is ethanesulfonate crystal form A, and an X-ray powder diffraction pattern of the ethanesulfonate crystal form A has a diffraction peak at 17.3 +/-0.2 degrees, or a diffraction peak at 19.5 +/-0.2 degrees, or a diffraction peak at 21.6 +/-0.2 degrees, or a diffraction peak at 18.5 +/-0.2 degrees, or a diffraction peak at 23.9 +/-0.2 degrees, or a diffraction peak at 10.6 +/-0.2 degrees, or a diffraction peak at 22.2 +/-0.2 degrees; or a diffraction peak at 13.9 + -0.2 deg., or a diffraction peak at 26.1 + -0.2 deg., or a diffraction peak at 12.3 + -0.2 deg., preferably including any 2-5, or 3-6, or 3-8, or 5-8, or 6-8, more preferably including any 6, 7, or 8 of the above diffraction peaks,
Preferably, the form a of the ethanesulfonate salt has an X-ray powder diffraction pattern comprising at least one diffraction peak, preferably 2, more preferably 3, and optionally further comprising at least one diffraction peak, preferably 2, 3, 4 or 5, having a 2 Θ of 17.3 ± 0.2 °, 19.5 ± 0.2 °, 21.6 ± 0.2 °, preferably 2, more preferably 3, and optionally further comprising at least one diffraction peak, preferably 2, 3, 4 or 5, having a 2 Θ of 18.5 ± 0.2 °, 23.9 ± 0.2 °, 10.6 ± 0.2 °, 22.2 ± 0.2 °, 13.9 ± 0.2 °,
for example, the ethane sulfonate form a has an X-ray powder diffraction pattern with diffraction peaks at the following positions 2 θ:
17.3 +/-0.2 degrees and 19.5 +/-0.2 degrees,
or, at 19.5 + -0.2 deg. and 21.6 + -0.2 deg.,
or 17.3 +/-0.2 degrees, 19.5 +/-0.2 degrees and 21.6 +/-0.2 degrees,
or at 19.5 +/-0.2 degrees, 21.6 +/-0.2 degrees and 18.5 +/-0.2 degrees,
or 17.3 +/-0.2 degrees, 19.5 +/-0.2 degrees, 21.6 +/-0.2 degrees and 18.5 +/-0.2 degrees,
or 19.5 +/-0.2 degrees, 21.6 +/-0.2 degrees, 18.5 +/-0.2 degrees and 23.9 +/-0.2 degrees,
or 17.3 +/-0.2 degrees, 19.5 +/-0.2 degrees, 21.6 +/-0.2 degrees, 18.5 +/-0.2 degrees and 23.9 +/-0.2 degrees,
or 19.5 +/-0.2 degrees, 21.6 +/-0.2 degrees, 18.5 +/-0.2 degrees, 23.9 +/-0.2 degrees and 10.6 +/-0.2 degrees,
more preferably, the X-ray powder diffraction pattern of ethane sulfonate form A optionally further comprises one or more diffraction peaks at 26.1 + -0.2 °, 12.3 + -0.2 °, 20.7 + -0.2 °, 21.3 + -0.2 °, 19.0 + -0.2 °, 28.1 + -0.2 °, 13.6 + -0.2 ° 2 θ, preferably at least any 2-3, or 4-5, or 6-7 thereof, further preferably any 2, 3, 4, 5, 6, 7 thereof,
For example, the ethane sulfonate form a has an X-ray powder diffraction pattern with diffraction peaks at the following positions 2 θ:
26.1 +/-0.2 degrees, 12.3 +/-0.2 degrees, 20.7 +/-0.2 degrees, 21.3 +/-0.2 degrees, 19.0 +/-0.2 degrees and 28.1 +/-0.2 degrees,
or 12.3 +/-0.2 degrees, 20.7 +/-0.2 degrees, 21.3 +/-0.2 degrees, 19.0 +/-0.2 degrees, 28.1 +/-0.2 degrees and 13.6 +/-0.2 degrees,
or 26.1 +/-0.2 degrees, 12.3 +/-0.2 degrees, 21.3 +/-0.2 degrees, 19.0 +/-0.2 degrees, 28.1 +/-0.2 degrees and 13.6 +/-0.2 degrees,
further preferably, the form a of the ethanesulfonate salt has an X-ray powder diffraction pattern comprising diffraction peaks at one or more of 17.3 ± 0.2 °, 19.5 ± 0.2 °, 21.6 ± 0.2 °, 18.5 ± 0.2 °, 23.9 ± 0.2 °, 10.6 ± 0.2 °, 22.2 ± 0.2 °, 13.9 ± 0.2 °, 26.1 ± 0.2 °, 12.3 ± 0.2 °, 20.7 ± 0.2 °, 21.3 ± 0.2 °, 19.0 ± 0.2 °, 28.1 ± 0.2 °, 13.6 ± 0.2 ° 2 Θ, preferably comprising diffraction peaks at any of 4, 5, 6, 8, 10,
for example, the ethane sulfonate form a has an X-ray powder diffraction pattern with diffraction peaks at the following positions 2 θ:
17.3 +/-0.2 degrees, 21.6 +/-0.2 degrees, 18.5 +/-0.2 degrees and 23.9 +/-0.2 degrees,
or 19.5 +/-0.2 degrees, 23.9 +/-0.2 degrees, 10.6 +/-0.2 degrees and 22.2 +/-0.2 degrees,
or 21.6 +/-0.2 degrees, 23.9 +/-0.2 degrees, 10.6 +/-0.2 degrees, 22.2 +/-0.2 degrees and 13.9 +/-0.2 degrees,
Or 18.5 +/-0.2 degrees, 10.6 +/-0.2 degrees, 22.2 +/-0.2 degrees, 13.9 +/-0.2 degrees and 26.1 +/-0.2 degrees,
or 17.3 +/-0.2 degrees, 19.5 +/-0.2 degrees, 21.6 +/-0.2 degrees, 18.5 +/-0.2 degrees, 23.9 +/-0.2 degrees and 13.9 +/-0.2 degrees,
or 19.5 +/-0.2 degrees, 21.6 +/-0.2 degrees, 18.5 +/-0.2 degrees, 23.9 +/-0.2 degrees, 10.6 +/-0.2 degrees and 26.1 +/-0.2 degrees,
or 21.6 +/-0.2 degrees, 18.5 +/-0.2 degrees, 23.9 +/-0.2 degrees, 10.6 +/-0.2 degrees, 22.2 +/-0.2 degrees, 13.9 +/-0.2 degrees, 26.1 +/-0.2 degrees and 12.3 +/-0.2 degrees,
or 18.5 +/-0.2 degrees, 23.9 +/-0.2 degrees, 10.6 +/-0.2 degrees, 22.2 +/-0.2 degrees, 13.9 +/-0.2 degrees, 26.1 +/-0.2 degrees, 12.3 +/-0.2 degrees and 20.7 +/-0.2 degrees,
or 17.3 +/-0.2 degrees, 19.5 +/-0.2 degrees, 21.6 +/-0.2 degrees, 18.5 +/-0.2 degrees, 23.9 +/-0.2 degrees, 10.6 +/-0.2 degrees, 22.2 +/-0.2 degrees, 13.9 +/-0.2 degrees, 26.1 +/-0.2 degrees and 12.3 +/-0.2 degrees,
or 19.5 +/-0.2 degrees, 21.6 +/-0.2 degrees, 18.5 +/-0.2 degrees, 23.9 +/-0.2 degrees, 10.6 +/-0.2 degrees, 22.2 +/-0.2 degrees, 13.9 +/-0.2 degrees, 26.1 +/-0.2 degrees, 12.3 +/-0.2 degrees and 20.7 +/-0.2 degrees,
still further preferably, the ethanesulfonate salt form a has an X-ray powder diffraction pattern as shown in fig. 33, or a DSC pattern as shown in fig. 34;
the succinate crystal form is succinate crystal form A, the X-ray powder diffraction pattern of the succinate crystal form A has diffraction peaks at 22.8 +/-0.2 degrees, or a diffraction peak at 19.9 + -0.2 deg., or a diffraction peak at 18.3 + -0.2 deg., or a diffraction peak at 12.3 + -0.2 deg., or a diffraction peak at 21.7 + -0.2 deg., or a diffraction peak at 24.7 + -0.2 deg., or a diffraction peak at 11.0 + -0.2 deg., or a diffraction peak at 15.2 + -0.2 deg., or a diffraction peak at 27.4 + -0.2 deg., or has a diffraction peak at 20.2 + -0.2 deg., preferably contains any 2-5 of the above diffraction peaks, or 3-5, or 3-6, or 3-8, or 5-8, or 6-8, more preferably including any 6, 7, or 8 thereof,
Preferably, the form a of the succinate salt has an X-ray powder diffraction pattern comprising at least one or more diffraction peaks, preferably 2, more preferably 3, optionally further comprising at least one of 12.3 ± 0.2 °, 21.7 ± 0.2 °, 24.7 ± 0.2 °, 11.0 ± 0.2 °, 15.2 ± 0.2 °, preferably 2, 3, 4 or 5, 2 Θ,
for example, the succinate form a has an X-ray powder diffraction pattern with diffraction peaks at the following positions 2 θ:
22.8 +/-0.2 degrees and 19.9 +/-0.2 degrees,
or, at 19.9 + -0.2 deg. and 18.3 + -0.2 deg.,
or at 22.8 +/-0.2 degrees, 19.9 +/-0.2 degrees and 18.3 +/-0.2 degrees,
or at 19.9 +/-0.2 degrees, 18.3 +/-0.2 degrees and 12.3 +/-0.2 degrees,
or 22.8 +/-0.2 degrees, 19.9 +/-0.2 degrees, 18.3 +/-0.2 degrees and 12.3 +/-0.2 degrees,
or 19.9 +/-0.2 degrees, 18.3 +/-0.2 degrees, 12.3 +/-0.2 degrees and 21.7 +/-0.2 degrees,
or 22.8 +/-0.2 degrees, 19.9 +/-0.2 degrees, 18.3 +/-0.2 degrees, 12.3 +/-0.2 degrees and 21.7 +/-0.2 degrees,
or 19.9 +/-0.2 degrees, 18.3 +/-0.2 degrees, 12.3 +/-0.2 degrees, 21.7 +/-0.2 degrees and 24.7 +/-0.2 degrees,
more preferably, the X-ray powder diffraction pattern of form a of the succinate salt optionally further comprises one or more diffraction peaks, preferably at least any 2-3, or 4-5, or 6-7, preferably further comprising any 2, 3, 4, 5, 6, 7, located at 2 Θ of 27.4 ± 0.2 °, 20.2 ± 0.2 °, 14.3 ± 0.2 °, 20.9 ± 0.2 °, 28.5 ± 0.2 °, 32.2 ± 0.2 °, 13.7 ± 0.2 °, more preferably at least any 2-3, or 4-5, or 6-7,
For example, the succinate form a has an X-ray powder diffraction pattern with diffraction peaks at the following positions 2 θ:
27.4 +/-0.2 degrees, 20.2 +/-0.2 degrees, 14.3 +/-0.2 degrees, 20.9 +/-0.2 degrees, 28.5 +/-0.2 degrees and 32.2 +/-0.2 degrees,
or 20.2 +/-0.2 degrees, 14.3 +/-0.2 degrees, 20.9 +/-0.2 degrees, 28.5 +/-0.2 degrees, 32.2 +/-0.2 degrees and 13.7 +/-0.2 degrees,
or 27.4 +/-0.2 degrees, 20.2 +/-0.2 degrees, 20.9 +/-0.2 degrees, 28.5 +/-0.2 degrees, 32.2 +/-0.2 degrees and 13.7 +/-0.2 degrees,
further preferably, the form a of the succinate salt has an X-ray powder diffraction pattern comprising diffraction peaks at one or more of 22.8 ± 0.2 °, 19.9 ± 0.2 °, 18.3 ± 0.2 °, 12.3 ± 0.2 °, 21.7 ± 0.2 °, 24.7 ± 0.2 °, 11.0 ± 0.2 °, 15.2 ± 0.2 °, 27.4 ± 0.2 °, 20.2 ± 0.2 °, 14.3 ± 0.2 °, 20.9 ± 0.2 °, 28.5 ± 0.2 °, 32.2 ± 0.2 °, 13.7 ± 0.2 ° 2 Θ, preferably comprising diffraction peaks at 4, 5, 6, 8, 10 optionally,
for example, the succinate form a has an X-ray powder diffraction pattern with diffraction peaks at the following positions 2 θ:
22.8 +/-0.2 degrees, 12.3 +/-0.2 degrees, 21.7 +/-0.2 degrees and 24.7 +/-0.2 degrees,
or 19.9 +/-0.2 degrees, 21.7 +/-0.2 degrees, 24.7 +/-0.2 degrees and 11.0 +/-0.2 degrees,
or 18.3 +/-0.2 degrees, 12.3 +/-0.2 degrees, 21.7 +/-0.2 degrees, 24.7 +/-0.2 degrees and 11.0 +/-0.2 degrees,
Or 12.3 +/-0.2 degrees, 21.7 +/-0.2 degrees, 24.7 +/-0.2 degrees, 11.0 +/-0.2 degrees and 15.2 +/-0.2 degrees,
or 22.8 +/-0.2 degrees, 19.9 +/-0.2 degrees, 18.3 +/-0.2 degrees, 12.3 +/-0.2 degrees, 21.7 +/-0.2 degrees and 15.2 +/-0.2 degrees,
or 19.9 +/-0.2 degrees, 18.3 +/-0.2 degrees, 12.3 +/-0.2 degrees, 21.7 +/-0.2 degrees, 24.7 +/-0.2 degrees and 14.3 +/-0.2 degrees,
or 18.3 +/-0.2 degrees, 12.3 +/-0.2 degrees, 21.7 +/-0.2 degrees, 24.7 +/-0.2 degrees, 11.0 +/-0.2 degrees, 15.2 +/-0.2 degrees, 27.4 +/-0.2 degrees and 20.2 +/-0.2 degrees,
or 12.3 +/-0.2 degrees, 21.7 +/-0.2 degrees, 24.7 +/-0.2 degrees, 11.0 +/-0.2 degrees, 15.2 +/-0.2 degrees, 27.4 +/-0.2 degrees, 20.2 +/-0.2 degrees and 14.3 +/-0.2 degrees,
or 22.8 +/-0.2 degrees, 19.9 +/-0.2 degrees, 18.3 +/-0.2 degrees, 12.3 +/-0.2 degrees, 21.7 +/-0.2 degrees, 24.7 +/-0.2 degrees, 11.0 +/-0.2 degrees, 15.2 +/-0.2 degrees, 27.4 +/-0.2 degrees and 11.0 +/-0.2 degrees,
or 19.9 +/-0.2 degrees, 18.3 +/-0.2 degrees, 12.3 +/-0.2 degrees, 21.7 +/-0.2 degrees, 24.7 +/-0.2 degrees, 11.0 +/-0.2 degrees, 15.2 +/-0.2 degrees, 27.4 +/-0.2 degrees, 20.2 +/-0.2 degrees and 14.3 +/-0.2 degrees,
still more preferably, succinate form a has an X-ray powder diffraction pattern as shown in figure 35, or a DSC pattern as shown in figure 36;
the crystal form of the hydrobromide is a crystal form A of the hydrobromide, the X-ray powder diffraction pattern of the crystal form A of the hydrobromide has a diffraction peak at 21.9 +/-0.2 degrees, or a diffraction peak at 20.3 + -0.2 deg., or a diffraction peak at 18.3 + -0.2 deg., or a diffraction peak at 12.0 + -0.2 deg., or a diffraction peak at 24.6 + -0.2 deg., or a diffraction peak at 16.8 + -0.2 deg., or a diffraction peak at 19.7 + -0.2 deg., or a diffraction peak at 21.4 + -0.2 deg., or a diffraction peak at 14.2 + -0.2 deg., or a diffraction peak at 32.2 + -0.2 deg., preferably including any 2-5 of the above diffraction peaks, or 3-5, or 3-6, or 3-8, or 5-8, or 6-8, more preferably including any 6, 7, or 8 thereof,
Preferably, the X-ray powder diffraction pattern of the hydrobromide form A comprises at least one diffraction peak at one or more of 21.9 + -0.2 deg., 20.3 + -0.2 deg., 18.3 + -0.2 deg., preferably 2 deg., more preferably 3 deg., optionally, further at least one of 12.0 + -0.2 deg., 24.6 + -0.2 deg., 16.8 + -0.2 deg., 19.7 + -0.2 deg., 21.4 + -0.2 deg., preferably 2 deg., 3 deg., 4 deg. or 5 deg. of 2 theta,
for example, the X-ray powder diffraction pattern of the hydrobromide form a has diffraction peaks at the following positions 2 θ:
21.9 +/-0.2 degrees and 20.3 +/-0.2 degrees,
or 20.3 +/-0.2 degrees and 18.3 +/-0.2 degrees,
or 21.9 +/-0.2 degrees, 20.3 +/-0.2 degrees and 18.3 +/-0.2 degrees,
or 20.3 +/-0.2 degrees, 18.3 +/-0.2 degrees and 12.0 +/-0.2 degrees,
or 21.9 +/-0.2 degrees, 20.3 +/-0.2 degrees, 18.3 +/-0.2 degrees and 12.0 +/-0.2 degrees,
or 20.3 +/-0.2 degrees, 18.3 +/-0.2 degrees, 12.0 +/-0.2 degrees and 24.6 +/-0.2 degrees,
or 21.9 +/-0.2 degrees, 20.3 +/-0.2 degrees, 18.3 +/-0.2 degrees, 12.0 +/-0.2 degrees and 24.6 +/-0.2 degrees,
or 20.3 +/-0.2 degrees, 18.3 +/-0.2 degrees, 12.0 +/-0.2 degrees, 24.6 +/-0.2 degrees and 16.8 +/-0.2 degrees,
more preferably, the X-ray powder diffraction pattern of crystalline hydrobromide form A optionally further comprises one or more diffraction peaks, preferably at least any 2-3, or 4-5, or 6-7 diffraction peaks, preferably at least any 2, 3, 4, 5, 6, 7 diffraction peaks, at 2 θ of 14.2 ± 0.2 °, 32.2 ± 0.2 °, 17.5 ± 0.2 °, 22.4 ± 0.2 °, 25.8 ± 0.2 °, 12.4 ± 0.2 °, 13.0 ± 0.2 °, further preferably, any 2, 3, 4, 5, 6, 7,
For example, the X-ray powder diffraction pattern of the hydrobromide form a has diffraction peaks at the following positions 2 θ:
14.2 +/-0.2 degrees, 32.2 +/-0.2 degrees, 17.5 +/-0.2 degrees, 22.4 +/-0.2 degrees, 25.8 +/-0.2 degrees and 12.4 +/-0.2 degrees,
or 32.2 +/-0.2 degrees, 17.5 +/-0.2 degrees, 22.4 +/-0.2 degrees, 25.8 +/-0.2 degrees, 12.4 +/-0.2 degrees and 13.0 +/-0.2 degrees,
or 14.2 +/-0.2 degrees, 32.2 +/-0.2 degrees, 22.4 +/-0.2 degrees, 25.8 +/-0.2 degrees, 12.4 +/-0.2 degrees and 13.0 +/-0.2 degrees,
further preferably, the X-ray powder diffraction pattern of hydrobromide form a comprises diffraction peaks at one or more of 21.9 ± 0.2 °, 20.3 ± 0.2 °, 18.3 ± 0.2 °, 12.0 ± 0.2 °, 24.6 ± 0.2 °, 16.8 ± 0.2 °, 19.7 ± 0.2 °, 21.4 ± 0.2 °, 14.2 ± 0.2 °, 32.2 ± 0.2 °, 17.5 ± 0.2 °, 22.4 ± 0.2 °, 25.8 ± 0.2 °, 12.4 ± 0.2 °, 13.0 ± 0.2 ° 2 Θ, preferably comprising optional diffraction peaks at 4, 5, 6, 8, 10 positions,
for example, the X-ray powder diffraction pattern of the hydrobromide form a has diffraction peaks at the following positions 2 θ:
21.9 +/-0.2 degrees, 18.3 +/-0.2 degrees, 12.0 +/-0.2 degrees and 24.6 +/-0.2 degrees,
or 20.3 +/-0.2 degrees, 24.6 +/-0.2 degrees, 16.8 +/-0.2 degrees and 19.7 +/-0.2 degrees,
or 18.3 +/-0.2 degrees, 12.0 +/-0.2 degrees, 24.6 +/-0.2 degrees, 16.8 +/-0.2 degrees and 21.4 +/-0.2 degrees,
Or 12.0 +/-0.2 degrees, 24.6 +/-0.2 degrees, 16.8 +/-0.2 degrees, 19.7 +/-0.2 degrees and 14.2 +/-0.2 degrees,
or 21.9 +/-0.2 degrees, 20.3 +/-0.2 degrees, 18.3 +/-0.2 degrees, 12.0 +/-0.2 degrees, 16.8 +/-0.2 degrees and 19.7 +/-0.2 degrees,
or 20.3 +/-0.2 degrees, 12.0 +/-0.2 degrees, 24.6 +/-0.2 degrees, 16.8 +/-0.2 degrees, 19.7 +/-0.2 degrees and 32.2 +/-0.2 degrees,
or 18.3 +/-0.2 degrees, 12.0 +/-0.2 degrees, 24.6 +/-0.2 degrees, 16.8 +/-0.2 degrees, 19.7 +/-0.2 degrees, 21.4 +/-0.2 degrees, 14.2 +/-0.2 degrees and 17.5 +/-0.2 degrees,
or 12.0 +/-0.2 degrees, 24.6 +/-0.2 degrees, 16.8 +/-0.2 degrees, 19.7 +/-0.2 degrees, 21.4 +/-0.2 degrees, 14.2 +/-0.2 degrees, 32.2 +/-0.2 degrees and 17.5 +/-0.2 degrees,
or 21.9 +/-0.2 degrees, 20.3 +/-0.2 degrees, 18.3 +/-0.2 degrees, 12.0 +/-0.2 degrees, 24.6 +/-0.2 degrees, 16.8 +/-0.2 degrees, 19.7 +/-0.2 degrees, 21.4 +/-0.2 degrees, 14.2 +/-0.2 degrees and 32.2 +/-0.2 degrees,
or 20.3 +/-0.2 degrees, 18.3 +/-0.2 degrees, 12.0 +/-0.2 degrees, 24.6 +/-0.2 degrees, 16.8 +/-0.2 degrees, 19.7 +/-0.2 degrees, 21.4 +/-0.2 degrees, 14.2 +/-0.2 degrees, 32.2 +/-0.2 degrees and 17.5 +/-0.2 degrees,
still further preferably, the hydrobromide form A has an X-ray powder diffraction pattern as shown in figure 37, or a DSC pattern as shown in figure 38;
the hydrochloride form is hydrochloride form A, the X-ray powder diffraction pattern of the hydrochloride form A has a diffraction peak at 22.5 +/-0.2 degrees, or a diffraction peak at 20.2 +/-0.2 degrees, or a diffraction peak at 12.2 +/-0.2 degrees, or a diffraction peak at 17.0 +/-0.2 degrees, or a diffraction peak at 28.7 +/-0.2 degrees, or a diffraction peak at 21.4 +/-0.2 degrees, or a diffraction peak at 18.7 +/-0.2 degrees, or a diffraction peak at 14.5 +/-0.2 degrees, or a diffraction peak at 18.0 +/-0.2 degrees, or a diffraction peak at 25.5 +/-0.2 degrees, preferably comprises any 2-5, or 3-6, or 3-8, or 5-8, or 6-8, more preferably comprises any 6, 7 or 8 therein,
Preferably, the X-ray powder diffraction pattern of the hydrochloride form A comprises at least one or more diffraction peaks, preferably 2, more preferably 3, with 2 theta being 22.5 + -0.2, 20.2 + -0.2 and 12.2 + -0.2, optionally further comprising at least one of 17.0 + -0.2, 28.7 + -0.2, 21.4 + -0.2, 18.7 + -0.2 and 14.5 + -0.2, preferably 2, 3, 4 or 5, with 2 theta,
for example, the X-ray powder diffraction pattern of the hydrochloride form a has diffraction peaks at the following positions 2 θ:
22.5 plus or minus 0.2 degrees and 20.2 plus or minus 0.2 degrees,
or 20.2 +/-0.2 degrees and 12.2 +/-0.2 degrees,
or at 22.5 +/-0.2 degrees, 20.2 +/-0.2 degrees and 12.2 +/-0.2 degrees,
or 20.2 +/-0.2 degrees, 12.2 +/-0.2 degrees and 17.0 +/-0.2 degrees,
or 22.5 +/-0.2 degrees, 20.2 +/-0.2 degrees, 12.2 +/-0.2 degrees and 17.0 +/-0.2 degrees,
or 20.2 +/-0.2 degrees, 12.2 +/-0.2 degrees, 17.0 +/-0.2 degrees and 28.7 +/-0.2 degrees,
or 22.5 +/-0.2 degrees, 20.2 +/-0.2 degrees, 12.2 +/-0.2 degrees, 17.0 +/-0.2 degrees and 28.7 +/-0.2 degrees,
or 20.2 +/-0.2 degrees, 12.2 +/-0.2 degrees, 17.0 +/-0.2 degrees, 28.7 +/-0.2 degrees and 21.4 +/-0.2 degrees,
more preferably, the X-ray powder diffraction pattern of form A of the hydrochloride salt optionally further comprises one or more diffraction peaks, preferably at least any 2-3, or 4-5, or 6-7 diffraction peaks, at 2, 3, 4, 5, 6, 7 diffraction peaks, at 2 θ of 18.0 ± 0.2 °, 25.5 ± 0.2 °, 28.3 ± 0.2 °, 23.8 ± 0.2 °, 12.6 ± 0.2 °, 24.8 ± 0.2 °, 13.3 ± 0.2 °, more preferably at least any 2-3, or 4, 5, 6, 7,
For example, the X-ray powder diffraction pattern of the hydrochloride form a has diffraction peaks at the following positions 2 θ:
18.0 +/-0.2 degrees, 25.5 +/-0.2 degrees, 28.3 +/-0.2 degrees, 23.8 +/-0.2 degrees, 12.6 +/-0.2 degrees and 24.8 +/-0.2 degrees,
or 25.5 +/-0.2 degrees, 28.3 +/-0.2 degrees, 23.8 +/-0.2 degrees, 12.6 +/-0.2 degrees, 24.8 +/-0.2 degrees and 13.3 +/-0.2 degrees,
or 18.0 +/-0.2 degrees, 25.5 +/-0.2 degrees, 23.8 +/-0.2 degrees, 12.6 +/-0.2 degrees, 24.8 +/-0.2 degrees and 13.3 +/-0.2 degrees,
further preferably, the X-ray powder diffraction pattern of the hydrochloride form a comprises diffraction peaks at one or more of 22.5 ± 0.2 °, 20.2 ± 0.2 °, 12.2 ± 0.2 °, 17.0 ± 0.2 °, 28.7 ± 0.2 °, 21.4 ± 0.2 °, 18.7 ± 0.2 °, 14.5 ± 0.2 °, 18.0 ± 0.2 °, 25.5 ± 0.2 °, 28.3 ± 0.2 °, 23.8 ± 0.2 °, 12.6 ± 0.2 °, 24.8 ± 0.2 °, 13.3 ± 0.2 ° of 2 Θ; preferably, the compound contains diffraction peaks at 4, 5, 6, 8 and 10,
for example, the X-ray powder diffraction pattern of the hydrochloride form a has diffraction peaks at the following positions 2 θ:
22.5 +/-0.2 degrees, 12.2 +/-0.2 degrees, 17.0 +/-0.2 degrees and 28.7 +/-0.2 degrees,
or 20.2 +/-0.2 degrees, 17.0 +/-0.2 degrees, 28.7 +/-0.2 degrees and 21.4 +/-0.2 degrees,
or 12.2 +/-0.2 degrees, 28.7 +/-0.2 degrees, 21.4 +/-0.2 degrees, 18.7 +/-0.2 degrees and 14.5 +/-0.2 degrees,
Or 17.0 +/-0.2 degrees, 28.7 +/-0.2 degrees, 21.4 +/-0.2 degrees, 18.7 +/-0.2 degrees and 14.5 +/-0.2 degrees,
or 22.5 +/-0.2 degrees, 12.2 +/-0.2 degrees, 17.0 +/-0.2 degrees, 28.7 +/-0.2 degrees, 21.4 +/-0.2 degrees and 18.7 +/-0.2 degrees,
or 20.2 +/-0.2 degrees, 17.0 +/-0.2 degrees, 28.7 +/-0.2 degrees, 21.4 +/-0.2 degrees, 18.7 +/-0.2 degrees and 14.5 +/-0.2 degrees,
or 12.2 +/-0.2 degrees, 17.0 +/-0.2 degrees, 28.7 +/-0.2 degrees, 21.4 +/-0.2 degrees, 18.7 +/-0.2 degrees, 14.5 +/-0.2 degrees, 18.0 +/-0.2 degrees and 25.5 +/-0.2 degrees,
or 17.0 +/-0.2 degrees, 28.7 +/-0.2 degrees, 21.4 +/-0.2 degrees, 18.7 +/-0.2 degrees, 14.5 +/-0.2 degrees, 18.0 +/-0.2 degrees, 25.5 +/-0.2 degrees and 28.3 +/-0.2 degrees,
or 22.5 +/-0.2 degrees, 20.2 +/-0.2 degrees, 12.2 +/-0.2 degrees, 17.0 +/-0.2 degrees, 28.7 +/-0.2 degrees, 21.4 +/-0.2 degrees, 18.7 +/-0.2 degrees, 14.5 +/-0.2 degrees, 18.0 +/-0.2 degrees and 25.5 +/-0.2 degrees,
or 20.2 +/-0.2 degrees, 12.2 +/-0.2 degrees, 17.0 +/-0.2 degrees, 28.7 +/-0.2 degrees, 21.4 +/-0.2 degrees, 18.7 +/-0.2 degrees, 14.5 +/-0.2 degrees, 18.0 +/-0.2 degrees, 25.5 +/-0.2 degrees and 28.3 +/-0.2 degrees,
still more preferably, the hydrochloride form a has an X-ray powder diffraction pattern as shown in figure 39, or a DSC pattern as shown in figure 40;
the maleate crystal form is a maleate crystal form A, the X-ray powder diffraction pattern of the maleate crystal form A has a diffraction peak at 21.4 +/-0.2 degrees, or a diffraction peak at 17.4 + -0.2 deg., or a diffraction peak at 20.0 + -0.2 deg., or a diffraction peak at 25.6 + -0.2 deg., or a diffraction peak at 13.8 + -0.2 deg., or a diffraction peak at 18.3 + -0.2 deg., or a diffraction peak at 16.2 + -0.2 deg., or a diffraction peak at 20.3 + -0.2 deg., or a diffraction peak at 22.9 + -0.2 deg., or a diffraction peak at 13.3 + -0.2 deg., preferably including any 2-5 of the above diffraction peaks, or 3-5, or 3-6, or 3-8, or 5-8, or 6-8, more preferably including any 6, 7, or 8 thereof,
Preferably, the X-ray powder diffraction pattern of the maleate form A comprises at least one or more diffraction peaks, preferably 2, more preferably 3, and optionally further at least one of 25.6 + -0.2, 13.8 + -0.2, 18.3 + -0.2, 16.2 + -0.2, and 20.3 + -0.2, preferably 2, 3, 4, or 5, 2 theta, 21.4 + -0.2, 20.0 theta,
for example, the maleate form a has an X-ray powder diffraction pattern with diffraction peaks at the following positions 2 θ:
21.4 +/-0.2 degrees and 17.4 +/-0.2 degrees,
or 17.4 +/-0.2 degrees and 20.0 +/-0.2 degrees,
or 21.4 +/-0.2 degrees, 17.4 +/-0.2 degrees and 20.0 +/-0.2 degrees,
or 17.4 +/-0.2 degrees, 20.0 +/-0.2 degrees and 25.6 +/-0.2 degrees,
or 21.4 +/-0.2 degrees, 17.4 +/-0.2 degrees, 20.0 +/-0.2 degrees and 25.6 +/-0.2 degrees,
or 17.4 +/-0.2 degrees, 20.0 +/-0.2 degrees, 25.6 +/-0.2 degrees and 13.8 +/-0.2 degrees,
or 21.4 +/-0.2 degrees, 17.4 +/-0.2 degrees, 20.0 +/-0.2 degrees, 25.6 +/-0.2 degrees and 13.8 +/-0.2 degrees,
or 17.4 +/-0.2 degrees, 20.0 +/-0.2 degrees, 25.6 +/-0.2 degrees, 13.8 +/-0.2 degrees and 18.3 +/-0.2 degrees,
more preferably, the X-ray powder diffraction pattern of maleate form A optionally further comprises one or more diffraction peaks at 22.9 + -0.2 °, 13.3 + -0.2 °, 22.5 + -0.2 °, 25.9 + -0.2 °, 10.9 + -0.2 °, 27.6 + -0.2 °, 23.5 + -0.2 °, preferably at least any 2-3, or 4-5, or 6-7, further preferably any 2, 3, 4, 5, 6, 7,
For example, the maleate form a has an X-ray powder diffraction pattern with diffraction peaks at the following positions 2 θ:
22.9 +/-0.2 degrees, 13.3 +/-0.2 degrees, 22.5 +/-0.2 degrees, 25.9 +/-0.2 degrees, 10.9 +/-0.2 degrees and 27.6 +/-0.2 degrees,
or 13.3 +/-0.2 degrees, 22.5 +/-0.2 degrees, 25.9 +/-0.2 degrees, 10.9 +/-0.2 degrees, 27.6 +/-0.2 degrees and 23.5 +/-0.2 degrees,
or 22.9 +/-0.2 degrees, 13.3 +/-0.2 degrees, 25.9 +/-0.2 degrees, 10.9 +/-0.2 degrees, 27.6 +/-0.2 degrees and 23.5 +/-0.2 degrees,
further preferably, the X-ray powder diffraction pattern of maleate form A comprises diffraction peaks at one or more of 21.4 + -0.2 °, 17.4 + -0.2 °, 20.0 + -0.2 °, 25.6 + -0.2 °, 13.8 + -0.2 °, 18.3 + -0.2 °, 16.2 + -0.2 °, 20.3 + -0.2 °, 22.9 + -0.2 °, 13.3 + -0.2 °, 22.5 + -0.2 °, 25.9 + -0.2 °, 10.9 + -0.2 °, 27.6 + -0.2 °, 23.5 + -0.2 ° of 2 θ, preferably comprising diffraction peaks at 4, 5, 6, 8, 10,
for example, the maleate form a has an X-ray powder diffraction pattern with diffraction peaks at the following positions 2 θ:
21.4 +/-0.2 degrees, 20.0 +/-0.2 degrees, 25.6 +/-0.2 degrees and 13.8 +/-0.2 degrees,
or 17.4 +/-0.2 degrees, 25.6 +/-0.2 degrees, 13.8 +/-0.2 degrees and 18.3 +/-0.2 degrees,
or 20.0 +/-0.2 degrees, 13.8 +/-0.2 degrees, 18.3 +/-0.2 degrees, 16.2 +/-0.2 degrees and 20.3 +/-0.2 degrees,
Or 25.6 +/-0.2 degrees, 18.3 +/-0.2 degrees, 16.2 +/-0.2 degrees, 20.3 +/-0.2 degrees and 22.9 +/-0.2 degrees,
or 21.4 +/-0.2 degrees, 20.0 +/-0.2 degrees, 25.6 +/-0.2 degrees, 13.8 +/-0.2 degrees, 18.3 +/-0.2 degrees and 16.2 +/-0.2 degrees,
or 17.4 +/-0.2 degrees, 25.6 +/-0.2 degrees, 13.8 +/-0.2 degrees, 18.3 +/-0.2 degrees, 16.2 +/-0.2 degrees and 20.3 +/-0.2 degrees,
or 20.0 +/-0.2 degrees, 13.8 +/-0.2 degrees, 18.3 +/-0.2 degrees, 16.2 +/-0.2 degrees, 20.3 +/-0.2 degrees, 22.9 +/-0.2 degrees, 13.3 +/-0.2 degrees and 22.5 +/-0.2 degrees,
or 25.6 +/-0.2 degrees, 18.3 +/-0.2 degrees, 16.2 +/-0.2 degrees, 20.3 +/-0.2 degrees, 22.9 +/-0.2 degrees, 13.3 +/-0.2 degrees, 22.5 +/-0.2 degrees and 25.9 +/-0.2 degrees,
or 21.4 +/-0.2 degrees, 17.4 +/-0.2 degrees, 20.0 +/-0.2 degrees, 25.6 +/-0.2 degrees, 13.8 +/-0.2 degrees, 18.3 +/-0.2 degrees, 16.2 +/-0.2 degrees, 20.3 +/-0.2 degrees, 22.9 +/-0.2 degrees and 13.3 +/-0.2 degrees,
or 17.4 +/-0.2 degrees, 20.0 +/-0.2 degrees, 25.6 +/-0.2 degrees, 13.8 +/-0.2 degrees, 18.3 +/-0.2 degrees, 16.2 +/-0.2 degrees, 20.3 +/-0.2 degrees, 22.9 +/-0.2 degrees, 13.3 +/-0.2 degrees and 22.5 +/-0.2 degrees,
still more preferably, maleate form a has an X-ray powder diffraction pattern as shown in figure 41, or a DSC pattern as shown in figure 42;
the tartrate crystal form is tartrate crystal form A, the X-ray powder diffraction pattern of the tartrate crystal form A has diffraction peaks at 20.1 +/-0.2 degrees, or a diffraction peak at 22.8 + -0.2 deg., or a diffraction peak at 12.6 + -0.2 deg., or has a diffraction peak at 18.4 + -0.2 deg., or has a diffraction peak at 18.1 + -0.2 deg., or a diffraction peak at 24.5 + -0.2 deg., or a diffraction peak at 11.2 + -0.2 deg., or a diffraction peak at 29.0 + -0.2 deg., or a diffraction peak at 20.7 + -0.2 deg., or a diffraction peak at 21.6 + -0.2 deg., preferably any 2-5 of the above diffraction peaks, or 3-5, or 3-6, or 3-8, or 5-8, or 6-8, more preferably including any 6, 7, or 8 thereof,
Preferably, the X-ray powder diffraction pattern of tartrate form A comprises at least one diffraction peak at one or more of 20.1 +/-0.2 degrees, 22.8 +/-0.2 degrees, 12.6 +/-0.2 degrees, preferably comprises 2 degrees, more preferably comprises 3 degrees, and optionally further comprises at least one of 18.4 +/-0.2 degrees, 18.1 +/-0.2 degrees, 24.5 +/-0.2 degrees, 11.2 +/-0.2 degrees, 29.0 +/-0.2 degrees, preferably comprises 2 degrees, 3 degrees, 4 degrees or 5 degrees of 2 theta,
for example, the tartrate form a has an X-ray powder diffraction pattern with diffraction peaks at the following positions 2 θ:
at 20.1 +/-0.2 degrees and 22.8 +/-0.2 degrees,
or at 22.8 +/-0.2 degrees and 12.6 +/-0.2 degrees,
or 20.1 +/-0.2 degrees, 22.8 +/-0.2 degrees and 12.6 +/-0.2 degrees,
or at 22.8 +/-0.2 degrees, 12.6 +/-0.2 degrees and 18.4 +/-0.2 degrees,
or 20.1 +/-0.2 degrees, 22.8 +/-0.2 degrees, 12.6 +/-0.2 degrees and 18.4 +/-0.2 degrees,
or at 22.8 +/-0.2 degrees, 12.6 +/-0.2 degrees, 18.4 +/-0.2 degrees and 18.1 +/-0.2 degrees,
or 20.1 +/-0.2 degrees, 22.8 +/-0.2 degrees, 12.6 +/-0.2 degrees, 18.4 +/-0.2 degrees and 18.1 +/-0.2 degrees,
or 22.8 +/-0.2 degrees, 12.6 +/-0.2 degrees, 18.4 +/-0.2 degrees, 18.1 +/-0.2 degrees and 24.5 +/-0.2 degrees,
more preferably, the X-ray powder diffraction pattern of tartrate form a optionally further comprises one or more diffraction peaks, preferably at least any 2 to 3, or 4 to 5, or 6 to 7, located at 20.7 ± 0.2 °, 21.6 ± 0.2 °, 33.7 ± 0.2 °, 22.1 ± 0.2 °, 13.9 ± 0.2 °, 15.3 ± 0.2 °, 27.4 ± 0.2 ° 2 Θ; further preferably, any 2, 3, 4, 5, 6, 7,
For example, the tartrate form a has an X-ray powder diffraction pattern with diffraction peaks at the following positions 2 θ:
20.7 +/-0.2 degrees, 21.6 +/-0.2 degrees, 33.7 +/-0.2 degrees, 22.1 +/-0.2 degrees, 13.9 +/-0.2 degrees and 15.3 +/-0.2 degrees,
or 21.6 +/-0.2 degrees, 33.7 +/-0.2 degrees, 22.1 +/-0.2 degrees, 13.9 +/-0.2 degrees, 15.3 +/-0.2 degrees and 27.4 +/-0.2 degrees,
or 20.7 +/-0.2 degrees, 21.6 +/-0.2 degrees, 33.7 +/-0.2 degrees, 13.9 +/-0.2 degrees, 15.3 +/-0.2 degrees and 27.4 +/-0.2 degrees,
further preferably, the X-ray powder diffraction pattern of tartrate form A comprises diffraction peaks at one or more of 20.1 + -0.2 °, 22.8 + -0.2 °, 12.6 + -0.2 °, 18.4 + -0.2 °, 18.1 + -0.2 °, 24.5 + -0.2 °, 11.2 + -0.2 °, 29.0 + -0.2 °, 20.7 + -0.2 °, 21.6 + -0.2 °, 33.7 + -0.2 °, 22.1 + -0.2 °, 13.9 + -0.2 °, 15.3 + -0.2 °, and 27.4 + -0.2 ° of 2 θ, preferably comprising diffraction peaks at 4, 5, 6, 8, and 10, optionally,
for example, the tartrate form a has an X-ray powder diffraction pattern with diffraction peaks at the following positions 2 θ:
20.1 +/-0.2 degrees, 12.6 +/-0.2 degrees, 18.4 +/-0.2 degrees and 18.1 +/-0.2 degrees,
or 22.8 +/-0.2 degrees, 18.4 +/-0.2 degrees, 18.1 +/-0.2 degrees and 24.5 +/-0.2 degrees,
or 12.6 +/-0.2 degrees, 24.5 +/-0.2 degrees, 11.2 +/-0.2 degrees, 29.0 +/-0.2 degrees and 20.7 +/-0.2 degrees,
Or 18.4 +/-0.2 degrees, 24.5 +/-0.2 degrees, 11.2 +/-0.2 degrees, 29.0 +/-0.2 degrees and 20.7 +/-0.2 degrees,
or 20.1 +/-0.2 degrees, 12.6 +/-0.2 degrees, 18.4 +/-0.2 degrees, 18.1 +/-0.2 degrees, 24.5 +/-0.2 degrees and 11.2 +/-0.2 degrees,
or 22.8 +/-0.2 degrees, 18.4 +/-0.2 degrees, 18.1 +/-0.2 degrees, 24.5 +/-0.2 degrees, 11.2 +/-0.2 degrees and 29.0 +/-0.2 degrees,
or 20.1 +/-0.2 degrees, 22.8 +/-0.2 degrees, 12.6 +/-0.2 degrees, 18.4 +/-0.2 degrees, 18.1 +/-0.2 degrees, 24.5 +/-0.2 degrees, 11.2 +/-0.2 degrees and 29.0 +/-0.2 degrees,
or 22.8 +/-0.2 degrees, 12.6 +/-0.2 degrees, 18.4 +/-0.2 degrees, 18.1 +/-0.2 degrees, 24.5 +/-0.2 degrees, 11.2 +/-0.2 degrees, 29.0 +/-0.2 degrees and 20.7 +/-0.2 degrees,
or 20.1 +/-0.2 degrees, 22.8 +/-0.2 degrees, 12.6 +/-0.2 degrees, 18.4 +/-0.2 degrees, 18.1 +/-0.2 degrees, 24.5 +/-0.2 degrees, 11.2 +/-0.2 degrees, 29.0 +/-0.2 degrees, 20.7 +/-0.2 degrees and 21.6 +/-0.2 degrees,
or 22.8 +/-0.2 degrees, 12.6 +/-0.2 degrees, 18.4 +/-0.2 degrees, 18.1 +/-0.2 degrees, 24.5 +/-0.2 degrees, 11.2 +/-0.2 degrees, 29.0 +/-0.2 degrees, 20.7 +/-0.2 degrees, 21.6 +/-0.2 degrees and 33.7 +/-0.2 degrees,
still more preferably, tartrate form a has an X-ray powder diffraction pattern, as shown in figure 43, or a DSC pattern, as shown in figure 44;
the crystalline form of sulfate is crystalline form A, which has an X-ray powder diffraction pattern with a diffraction peak at 21.6 + -0.2 DEG, or a diffraction peak at 23.8 + -0.2 DEG, or a diffraction peak at 17.5 + -0.2 DEG, or a diffraction peak at 19.7 + -0.2 DEG, or a diffraction peak at 19.0 + -0.2 DEG, or a diffraction peak at 10.5 + -0.2 DEG, or a diffraction peak at 26.1 + -0.2 DEG, or a diffraction peak at 18.5 + -0.2 DEG, or a diffraction peak at 22.5 + -0.2 DEG, or a diffraction peak at 13.5 + -0.2 DEG, preferably comprising any of the above diffraction peaks 2-5, or 3-6, or 3-8, or 5-8, or 6-8, more preferably comprising any 6, 7 or 8 therein,
Preferably, the X-ray powder diffraction pattern of the sulfate form A at least comprises one or more diffraction peaks with 2 theta of 21.6 +/-0.2 degrees, 23.8 +/-0.2 degrees and 17.5 +/-0.2 degrees, preferably comprises 2 degrees, more preferably comprises 3 degrees, and optionally further comprises at least one of 2 theta of 19.7 +/-0.2 degrees, 19.0 +/-0.2 degrees, 10.5 +/-0.2 degrees, 26.1 +/-0.2 degrees and 18.5 +/-0.2 degrees, preferably comprises 2 degrees, 3 degrees, 4 degrees or 5 degrees,
for example, the X-ray powder diffraction pattern of the sulfate form a has diffraction peaks at the following positions 2 θ:
21.6 +/-0.2 degrees and 23.8 +/-0.2 degrees,
or at 23.8 +/-0.2 degrees and 17.5 +/-0.2 degrees,
or 21.6 +/-0.2 degrees, 23.8 +/-0.2 degrees and 17.5 +/-0.2 degrees,
or at 23.8 +/-0.2 degrees, 17.5 +/-0.2 degrees and 19.7 +/-0.2 degrees,
or 21.6 +/-0.2 degrees, 23.8 +/-0.2 degrees, 17.5 +/-0.2 degrees and 19.7 +/-0.2 degrees,
or at 23.8 +/-0.2 degrees, 17.5 +/-0.2 degrees, 19.7 +/-0.2 degrees and 19.0 +/-0.2 degrees,
or 21.6 +/-0.2 degrees, 23.8 +/-0.2 degrees, 17.5 +/-0.2 degrees, 19.7 +/-0.2 degrees and 19.0 +/-0.2 degrees,
or at 23.8 + -0.2 deg., 17.5 + -0.2 deg., 19.7 + -0.2 deg., 19.0 + -0.2 deg. and 10.5 + -0.2 deg.,
more preferably, the X-ray powder diffraction pattern of form A of the sulfate salt optionally further comprises one or more diffraction peaks, preferably at least any 2-3, or 4-5, or 6-7, further preferably any 2, 3, 4, 5, 6, or 7, located at 22.5 + -0.2 °, 13.5 + -0.2 °, 14.0 + -0.2 °, 20.6 + -0.2 °, 12.3 + -0.2 °, 20.9 + -0.2 °, 31.7 + -0.2 ° 2 θ,
For example, the X-ray powder diffraction pattern of the sulfate form a has diffraction peaks at the following positions 2 θ:
22.5 +/-0.2 degrees, 13.5 +/-0.2 degrees, 14.0 +/-0.2 degrees, 20.6 +/-0.2 degrees, 12.3 +/-0.2 degrees and 20.9 +/-0.2 degrees,
or 13.5 +/-0.2 degrees, 14.0 +/-0.2 degrees, 20.6 +/-0.2 degrees, 12.3 +/-0.2 degrees, 20.9 +/-0.2 degrees and 31.7 +/-0.2 degrees,
or 22.5 +/-0.2 degrees, 13.5 +/-0.2 degrees, 20.6 +/-0.2 degrees, 12.3 +/-0.2 degrees, 20.9 +/-0.2 degrees and 31.7 +/-0.2 degrees,
further preferably, the X-ray powder diffraction pattern of the sulfate form A comprises diffraction peaks at one or more of 21.6 + -0.2 °, 23.8 + -0.2 °, 17.5 + -0.2 °, 19.7 + -0.2 °, 19.0 + -0.2 °, 10.5 + -0.2 °, 26.1 + -0.2 °, 18.5 + -0.2 °, 22.5 + -0.2 °, 13.5 + -0.2 °, 14.0 + -0.2 °, 20.6 + -0.2 °, 12.3 + -0.2 °, 20.9 + -0.2 °, and 31.7 + -0.2 ° of 2 θ, preferably, comprises diffraction peaks at 4, 5, 6, 8, and 10, optionally,
for example, the X-ray powder diffraction pattern of the sulfate form a has diffraction peaks at the following positions 2 θ:
21.6 +/-0.2 degrees, 17.5 +/-0.2 degrees, 19.7 +/-0.2 degrees and 19.0 +/-0.2 degrees,
or 23.8 +/-0.2 degrees, 19.7 +/-0.2 degrees, 19.0 +/-0.2 degrees and 10.5 +/-0.2 degrees,
or 17.5 +/-0.2 degrees, 19.0 +/-0.2 degrees, 10.5 +/-0.2 degrees, 26.1 +/-0.2 degrees and 18.5 +/-0.2 degrees,
Or 19.7 +/-0.2 degrees, 10.5 +/-0.2 degrees, 26.1 +/-0.2 degrees, 18.5 +/-0.2 degrees and 22.5 +/-0.2 degrees,
or 21.6 +/-0.2 degrees, 17.5 +/-0.2 degrees, 19.7 +/-0.2 degrees, 19.0 +/-0.2 degrees, 10.5 +/-0.2 degrees and 26.1 +/-0.2 degrees,
or 23.8 +/-0.2 degrees, 19.7 +/-0.2 degrees, 19.0 +/-0.2 degrees, 10.5 +/-0.2 degrees, 26.1 +/-0.2 degrees and 18.5 +/-0.2 degrees,
or 17.5 +/-0.2 degrees, 19.0 +/-0.2 degrees, 10.5 +/-0.2 degrees, 26.1 +/-0.2 degrees, 18.5 +/-0.2 degrees, 22.5 +/-0.2 degrees, 13.5 +/-0.2 degrees and 14.0 +/-0.2 degrees,
or 19.7 +/-0.2 degrees, 10.5 +/-0.2 degrees, 26.1 +/-0.2 degrees, 18.5 +/-0.2 degrees, 22.5 +/-0.2 degrees, 13.5 +/-0.2 degrees, 14.0 +/-0.2 degrees and 20.6 +/-0.2 degrees,
or 21.6 +/-0.2 degrees, 23.8 +/-0.2 degrees, 17.5 +/-0.2 degrees, 19.7 +/-0.2 degrees, 19.0 +/-0.2 degrees, 10.5 +/-0.2 degrees, 26.1 +/-0.2 degrees, 18.5 +/-0.2 degrees, 22.5 +/-0.2 degrees and 13.5 +/-0.2 degrees,
or 23.8 +/-0.2 degrees, 17.5 +/-0.2 degrees, 19.7 +/-0.2 degrees, 19.0 +/-0.2 degrees, 10.5 +/-0.2 degrees, 26.1 +/-0.2 degrees, 18.5 +/-0.2 degrees, 22.5 +/-0.2 degrees, 13.5 +/-0.2 degrees and 14.0 +/-0.2 degrees,
still further preferred form A of the sulfate salt has an X-ray powder diffraction pattern as shown in figure 45 or a DSC pattern as shown in figure 46.
16. A crystalline form according to any of claims 14-15, characterized in that the 2 Θ error of the diffraction peak position with the first ten strong relative peak intensity in the X-ray powder diffraction pattern of the compound (S) -1'- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) -3-fluoropyrazin-2-yl) -1, 3-dihydrospiro [ indene-2, 4' -piperidin ] -1-amine hydrochloride form a, ethanesulfonate form a from the diffraction peak position corresponding to fig. 1, fig. 6 is ± 0.2 ° -0.5 °, preferably ± 0.2-0.3 °, most preferably ± 0.2 °;
Or, the diffraction peak positions of the relative peak intensities of the mesylate crystal form A, the isethionate crystal form A, the fumarate crystal form A, the p-toluenesulfonate crystal form A, the ethanesulfonate crystal form A, the succinate crystal form A, the hydrobromide crystal form A, the hydrochloride crystal form A, the maleate crystal form A, the tartrate crystal form A and the sulfate crystal form A of the compound (S) -8- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) pyrazin-2-yl) -2-cyclopropyl-8-azaspiro [4.5] dec-2-en-1-amine in the X-ray powder diffraction patterns of the compound (S) -8- (6-amino-5- ((2-amino-3-chloropyridin-4-yl) thio) pyrazin-2-yl) -2-cyclopropyl-8-azaspiro [4.5] en-1-amine are first-intense as compared with the diffraction peak positions of FIG. 25, FIG. 27, FIG. 29, FIG. 31, FIG. 33, FIG. 35, FIG. 37, FIG. 39, and FIG. 39, The 2 θ errors of diffraction peaks at the corresponding positions in fig. 41, 43, and 45 are ± 0.2 ° to ± 0.5 °, preferably ± 0.2 ° to ± 0.3 °, and most preferably ± 0.2 °.
17. A process for preparing the acid salt of any one of claims 1 to 13 or the acid salt crystalline form of any one of claims 14 to 16, comprising the steps of:
1) weighing a proper amount of compound free alkali, dissolving the compound free alkali by a benign solvent,
2) dissolving the acid with an organic solvent; the amount of acid is preferably 2 equivalents,
3) mixing the two solutions obtained in the steps 1) and 2) to obtain a salt solution, stirring and precipitating or dripping a poor solvent and stirring and precipitating,
4) Quickly centrifuging or standing and drying to obtain a target product,
wherein:
the benign solvent of the step 1) is selected from methanol, ethanol, ethyl acetate, dichloromethane, acetone, acetonitrile, 2-butanone, 3-pentanone, tetrahydrofuran, N-dimethylformamide, N-dimethylacetamide or N-methylpyrrolidone; preferably a mixture of methanol or ethanol, preferably methanol or ethanol,
the organic solvent in the step 2) is selected from methanol, ethanol, ethyl acetate, dichloromethane, acetone, N-hexane, petroleum ether, benzene, toluene, chloroform, acetonitrile, carbon tetrachloride, dichloroethane, tetrahydrofuran, 2-butanone, 3-pentanone, heptane, methyl tert-butyl ether, isopropyl ether, 1, 4-dioxane, tert-butyl alcohol or N, N-dimethylformamide; preferably methanol, ethanol or acetonitrile, and is preferably used,
the benign solvent in the step 1) and the organic solution in the step 2) need to be mutually soluble when in use,
the poor solvent of the step 3) is selected from heptane, water, methyl tert-butyl ether, toluene, isopropyl ether, ethyl acetate, acetone or acetonitrile; preferably water, methyl tert-butyl ether or isopropyl ether;
alternatively, the first and second electrodes may be,
1) weighing a proper amount of free alkali, dissolving the free alkali in a benign solvent,
2) dissolving the acid with an organic solvent; the amount of acid is preferably 2 equivalents,
3) uniformly mixing the two solutions obtained in the steps 1) and 2), volatilizing, adding a poor solvent after the solvent is volatilized, stirring for crystallization,
4) Quickly centrifuging or standing and drying to obtain a target product,
wherein:
the benign solvent of the step 1) is selected from methanol, ethanol, ethyl acetate, dichloromethane, acetone, acetonitrile, 2-butanone, 3-pentanone, tetrahydrofuran, N-dimethylformamide, N-dimethylacetamide or N-methylpyrrolidone; preferably a mixture of methanol or ethanol, preferably methanol or ethanol,
the organic solvent in the step 2) is selected from methanol, ethanol, ethyl acetate, dichloromethane, acetone, N-hexane, petroleum ether, benzene, toluene, chloroform, acetonitrile, carbon tetrachloride, dichloroethane, tetrahydrofuran, 2-butanone, 3-pentanone, heptane, methyl tert-butyl ether, isopropyl ether, 1, 4-dioxane, tert-butyl alcohol or N, N-dimethylformamide; preferably methanol, ethanol or acetonitrile, and is preferably used,
the benign solvent in the step 1) and the organic solution in the step 2) need to be mutually soluble when in use,
the poor solvent of the step 3) is selected from ethyl acetate, acetone, dichloromethane, acetonitrile, tetrahydrofuran, 2-butanone, 3-pentanone and 1, 4-dioxane, preferably ethyl acetate, acetone or acetonitrile;
alternatively, the first and second electrodes may be,
1) weighing a proper amount of compound salt, and suspending with a poor solvent to obtain a suspension; the suspension density is preferably 50-100 mg/mL,
2) shaking the suspension obtained in the step 1) for 1-10 days at 25-40 ℃,
3) Quickly centrifuging the suspension obtained in the step 2), removing supernatant, drying to obtain a target product,
wherein:
the poor solvent of the step 1) is selected from acetone, ethyl acetate, acetonitrile, ethanol, 88% acetone, tetrahydrofuran, 1, 4-dioxane, benzene, toluene, isopropanol, N-butanol, isobutanol, N-dimethylformamide, N-dimethylacetamide, N-propanol, tert-butanol or 2-butanone;
alternatively, the first and second electrodes may be,
1) weighing a proper amount of compound salt, dissolving the compound salt by a benign solvent,
2) cooling the solution obtained in the step 1), stirring, precipitating solid, preferably selecting the temperature of-20-5 ℃,
3) quickly centrifuging the suspension obtained in the step 2), removing supernatant, drying to obtain a target product,
wherein:
the benign solvent is selected from methanol, ethanol, 88% acetone, tetrahydrofuran, N-dimethylformamide, N-dimethylacetamide or N-methylpyrrolidone; preferably methanol or 88% acetone;
alternatively, the first and second electrodes may be,
1) weighing a proper amount of free alkali, dissolving the free alkali in a benign solvent,
2) weighing a proper amount of acid, and dissolving the acid by using an organic solvent; the amount of acid is preferably 1.2 equivalents,
3) mixing the two solutions, stirring to separate out or dripping poor solvent and stirring to separate out,
4) Drying the mixture to obtain a target product,
wherein:
the benign solvent is selected from methanol, dichloromethane, 1, 4-dioxane, acetone, tetrahydrofuran, N-dimethylformamide, N-dimethylacetamide or N-methylpyrrolidone; preferably methanol or tetrahydrofuran, and preferably, methanol or tetrahydrofuran,
the organic solvent is selected from methanol, ethanol, ethyl acetate, dichloromethane, acetone, N-hexane, petroleum ether, benzene, toluene, chloroform, acetonitrile, carbon tetrachloride, dichloroethane, tetrahydrofuran, 2-butanone, 3-pentanone, heptane, methyl tert-butyl ether, isopropyl ether, 1, 4-dioxane, tert-butanol or N, N-dimethylformamide; preferably methanol, ethanol or acetonitrile; the benign solvent and the organic solution need to be mutually soluble when in use,
the poor solvent is selected from heptane, water, methyl tert-butyl ether, toluene, isopropyl ether, ethyl acetate, acetone or acetonitrile; preferably water, methyl tert-butyl ether, isopropyl ether;
alternatively, the first and second electrodes may be,
1) weighing a proper amount of free alkali, suspending with a poor solvent 1,
2) weighing a proper amount of acid, and dissolving the acid by using an organic solvent; the amount of acid is preferably 1.2 equivalents,
3) mixing the two solutions, stirring for dissolving, continuously stirring for precipitating or dripping the poor solvent 2 and stirring for precipitating,
4) drying the mixture to obtain a target product,
The poor solvent 1 is selected from ethyl acetate, acetone, dichloromethane, acetonitrile, tetrahydrofuran, 2-methyl-tetrahydrofuran, 2-butanone, 3-pentanone, 1, 4-dioxane and ethanol; preferably ethyl acetate, acetone, acetonitrile, 2-methyl-tetrahydrofuran, ethanol,
the organic solvent is selected from methanol, ethanol, ethyl acetate, dichloromethane, acetone, N-hexane, petroleum ether, benzene, toluene, chloroform, acetonitrile, carbon tetrachloride, dichloroethane, tetrahydrofuran, 2-butanone, 3-pentanone, heptane, methyl tert-butyl ether, isopropyl ether, 1, 4-dioxane, tert-butanol or N, N-dimethylformamide; preferably methanol, ethanol or acetonitrile; the benign solvent and the organic solution need to be mutually soluble when in use,
the poor solvent 2 is selected from heptane, water, methyl tert-butyl ether, toluene, isopropyl ether, ethyl acetate, acetone or acetonitrile; preferably water, methyl tert-butyl ether, isopropyl ether;
the acid is selected from hydrochloric acid, sulfuric acid, nitric acid, hydrobromic acid, hydrofluoric acid, hydroiodic acid, phosphoric acid, 2, 5-dihydroxybenzoic acid, 1-hydroxy-2-naphthoic acid, acetic acid, dichloroacetic acid, trichloroacetic acid, acetoxy-hydroxamic acid, adipic acid, benzenesulfonic acid, 4-chlorobenzenesulfonic acid, benzoic acid, 4-acetamidobenzoic acid, 4-aminobenzoic acid, capric acid, caproic acid, caprylic acid, cinnamic acid, citric acid, cyclamic acid, camphorsulfonic acid, aspartic acid, camphoric acid, gluconic acid, glucuronic acid, glutamic acid, isoascorbic acid, lactic acid, malic acid, mandelic acid, pyroglutamic acid, tartaric acid, dodecylsulfuric acid, dibenzoyltartaric acid, ethane-1, 2-disulfonic acid, ethanesulfonic acid, formic acid, fumaric acid, galactaric acid, gentisic acid, glutaric acid, 2-ketoglutaric acid, Glycolic acid, hippuric acid, isethionic acid, lactobionic acid, ascorbic acid, aspartic acid, lauric acid, camphoric acid, maleic acid, malonic acid, methanesulfonic acid, 1, 5-naphthalenedisulfonic acid, naphthalene-2-sulfonic acid, nicotinic acid, oleic acid, orotic acid, oxalic acid, palmitic acid, pamoic acid, propionic acid, salicylic acid, 4-aminosalicylic acid, sebacic acid, stearic acid, succinic acid, thiocyanic acid, undecylenic acid, trifluoroacetic acid, benzenesulfonic acid, p-toluenesulfonic acid, or L-malic acid; preferably fumaric acid, p-toluenesulfonic acid or succinic acid; most preferred is p-toluenesulfonic acid.
18. A pharmaceutical composition comprising a therapeutically effective amount of the acid salt of any one of claims 1-13 or the crystalline form of the acid salt of any one of claims 14-16, and one or more pharmaceutically acceptable carriers, diluents, or excipients.
19. Use of the acid salt according to any one of claims 1 to 13 or the acid salt crystalline form according to any one of claims 14 to 16 and the pharmaceutical composition according to claim 18 for the preparation of a SHP-2 inhibitor medicament.
20. The use according to claim 19, wherein the use is in the manufacture of a medicament for the treatment of noonan's syndrome, leopard skin syndrome, leukemia, neuroblastoma, melanoma, esophageal cancer, head and neck tumors, breast cancer, lung cancer, and colon cancer diseases or disorders thereof; preferably non-small cell lung cancer, esophageal cancer or head and neck tumors.
CN202110360835.8A 2020-04-03 2021-04-02 Salt of nitrogen-containing heteroaromatic derivative and crystal form thereof Pending CN113493440A (en)

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Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2022235864A1 (en) 2021-05-05 2022-11-10 Revolution Medicines, Inc. Ras inhibitors
WO2022235870A1 (en) 2021-05-05 2022-11-10 Revolution Medicines, Inc. Ras inhibitors for the treatment of cancer
WO2023051606A1 (en) * 2021-09-28 2023-04-06 上海翰森生物医药科技有限公司 Medical use of shp2 inhibitor in combination with egfr-tki in treatment and prevention of tumor diseases

Citations (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN105916845A (en) * 2014-01-17 2016-08-31 诺华股份有限公司 N-azaspirocycloalkane substituted n-heteroaryl compounds and compositions for inhibiting the activity of shp2
CN107922388A (en) * 2015-06-19 2018-04-17 诺华股份有限公司 For suppressing the compound and composition of SHP2 activity
WO2018172984A1 (en) * 2017-03-23 2018-09-27 Jacobio Pharmaceuticals Co., Ltd. Novel heterocyclic derivatives useful as shp2 inhibitors
CN109983001A (en) * 2016-07-12 2019-07-05 锐新医药公司 Tri- substituted type 3- methylpyrazine of the disubstituted type 3- methylpyrazine of 2,5- and 2,5,6- as allosteric SHP2 inhibitor
WO2019183367A1 (en) * 2018-03-21 2019-09-26 Relay Therapeutics, Inc. Shp2 phosphatase inhibitors and methods of use thereof
WO2020063760A1 (en) * 2018-09-26 2020-04-02 Jacobio Pharmaceuticals Co., Ltd. Novel heterocyclic derivatives useful as shp2 inhibitors
WO2020073949A1 (en) * 2018-10-10 2020-04-16 江苏豪森药业集团有限公司 Regulator of nitrogen-containing heteroaromatic derivatives, preparation method therefor and use thereof

Patent Citations (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN105916845A (en) * 2014-01-17 2016-08-31 诺华股份有限公司 N-azaspirocycloalkane substituted n-heteroaryl compounds and compositions for inhibiting the activity of shp2
CN107922388A (en) * 2015-06-19 2018-04-17 诺华股份有限公司 For suppressing the compound and composition of SHP2 activity
CN109983001A (en) * 2016-07-12 2019-07-05 锐新医药公司 Tri- substituted type 3- methylpyrazine of the disubstituted type 3- methylpyrazine of 2,5- and 2,5,6- as allosteric SHP2 inhibitor
WO2018172984A1 (en) * 2017-03-23 2018-09-27 Jacobio Pharmaceuticals Co., Ltd. Novel heterocyclic derivatives useful as shp2 inhibitors
WO2019183367A1 (en) * 2018-03-21 2019-09-26 Relay Therapeutics, Inc. Shp2 phosphatase inhibitors and methods of use thereof
WO2020063760A1 (en) * 2018-09-26 2020-04-02 Jacobio Pharmaceuticals Co., Ltd. Novel heterocyclic derivatives useful as shp2 inhibitors
WO2020073949A1 (en) * 2018-10-10 2020-04-16 江苏豪森药业集团有限公司 Regulator of nitrogen-containing heteroaromatic derivatives, preparation method therefor and use thereof

Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2022235864A1 (en) 2021-05-05 2022-11-10 Revolution Medicines, Inc. Ras inhibitors
WO2022235870A1 (en) 2021-05-05 2022-11-10 Revolution Medicines, Inc. Ras inhibitors for the treatment of cancer
WO2023051606A1 (en) * 2021-09-28 2023-04-06 上海翰森生物医药科技有限公司 Medical use of shp2 inhibitor in combination with egfr-tki in treatment and prevention of tumor diseases

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