Isotopologue of 2,4-disubstituted pyrimidine derivative and use thereof

Abstract

Disclosed in the present invention are an isotopologue of a 2,4-disubstituted pyrimidine derivative and a use thereof. Specifically, the present invention provides a compound as shown in formula (I), a solvate thereof, a pharmaceutically acceptable salt thereof, or a solvate of the pharmaceutically acceptable salt thereof. The compound provided by the present invention has good inhibitory activity against JAK and / or FLT kinases, and has good pharmacodynamic properties.

Description

Isotopes of 2,4-disubstituted pyrimidine derivatives and their uses

[0001] This application claims priority to Chinese patent application 2025104934653, filed on April 18, 2025. The entire contents of the aforementioned Chinese patent application are incorporated herein by reference. Technical Field

[0002] This invention belongs to the field of chemical medicine, specifically relating to isotopes of 2,4-disubstituted pyrimidine derivatives and their uses. Background Technology

[0003] Myeloproliferative neoplasms (MPNs) are a group of malignant myeloproliferative disorders originating from pluripotent hematopoietic stem cells. They are characterized by excessive proliferation of one or more myeloid cell lineages, leading to an increase in one or more lineages in the peripheral blood. These disorders tend to cause thrombosis, extramedullary hematopoiesis, myelofibrosis, and transformation into acute leukemia. These diseases include polycythemia vera (PV), primary thrombocytosis (PT), and primary myelofibrosis (PMF).

[0004] Current clinical treatments cannot cure myelofibrosis (MPNs). Recent studies have found that JAK2 kinase, a member of the Janus kinase (JAK) family, plays a crucial role in MPNs. The JAK-signal transducer and transcription activator-STAT (JAK-STAT) pathway controls the survival, proliferation, and differentiation of various cells through cytokine-mediated signal transduction. JAK2 phosphorylation, downstream STAT phosphorylation, and activation of gene transcription ultimately lead to increased proliferation, differentiation, and survival of erythrocytes and myeloid cells. Several JAK2 inhibitors are currently undergoing clinical trials. Among them, the JAK2 inhibitor Ruxolitinib has been approved by the FDA for myelofibrosis, while others such as Lestaurtinib (CEP701), CYT-387, LY2784544, and BMS-911543 are still in clinical research.

[0005] Furthermore, recent research indicates that FMS-like tyrosine kinase 3 (FLT3) mutations are also closely related to MPNs: knocking in FLT3 with an ITD (internal tandem repeat) mutation in mice can lead to myeloproliferative disorders. FLT3 is a receptor tyrosine kinase that plays a crucial role in the development of hematopoietic progenitor cells. Approximately 30% of patients with acute myeloid leukemia (AML) have activated FLT3 internal tandem repeat (ITD) mutations, a high-risk factor for disease relapse. Small molecule FLT3 inhibitors have been tested in clinical trials as single agents or in combination with chemotherapy; however, to date, these candidates have either failed to produce sufficient initial response or maintain therapeutic efficacy, primarily due to secondary resistance. Clinical data also show a sharp decline in peripheral blood leukemia cells after treatment, but with little bone marrow response. One possible mechanism for this failure is the existence of independent alternative survival pathways through which leukemia cells can adapt via further genetic mutations or metabolic adaptations. These pathways may include mTOR-PI3K-Akt, JAK-STAT, or Ras-MAPK. Simultaneously inhibiting these pathways may free leukemia cells from FLT3 restriction.

[0006] Building on this, simultaneously targeting the JAK2 pathway offers several advantages: (a) JAK2 mutations are rarely found in AML cases; (b) elevated phosphorylated JAK2 levels are observed in AML; and (c) in the FLT3-ITD strain, the negative regulators of JAK signaling, namely the inhibitors of cytokines 1 / 2 / 3, are significantly downregulated. Furthermore, there is evidence that inhibiting both the JAK2 and FLT3 signaling pathways can enhance clinical efficacy in AML patients with FLT3-ITD mutations. Based on this, JAK2 / FLT3 dual-target inhibitors for treating myelofibrosis are gradually becoming a research hotspot. Currently, the JAK2 / FLT3 dual-target inhibitor Fedratinib has been approved by the FDA for priority use in myelofibrosis, and the macrocyclic compound Pacritinib is also undergoing phase III clinical trials (for the treatment of myelofibrosis). However, existing JAK2 / FLT3 dual-target inhibitors have poor enzyme activity and low oral bioavailability, which still cannot meet medical needs. Therefore, developing inhibitors with better selectivity, higher activity, and better in vivo pharmacokinetic properties is currently a hot research topic.

[0007] The 2,4-disubstituted pyrimidine derivative compounds disclosed in patent WO2020259683A1 can be used as kinase inhibitors with dual targets of JAK2 and FLT3, or as kinase inhibitors with single targets of JAK2 or FLT3, providing new options for the preparation of drugs for the treatment and / or prevention of tumors, inflammatory diseases, and immune diseases.

[0008] Despite the beneficial activities of 2,4-disubstituted pyrimidine derivatives, there remains a persistent need for novel compounds to treat the aforementioned diseases and conditions. There is still a need in the art to develop compounds with JAK kinase inhibitory and FLT kinase inhibitory activities, as well as improved pharmacodynamic properties. Summary of the Invention

[0009] The technical problem this invention aims to solve is the limited variety of JAK kinase inhibitors or FLT kinase inhibitors in the prior art. Therefore, this invention provides isotopes of 2,4-disubstituted pyrimidine derivatives and their uses. The compounds provided by this invention exhibit good inhibitory activity against JAK kinase and / or FLT kinase, and also possess excellent pharmacodynamic properties.

[0010] This invention provides a compound of formula (I), a solvate thereof, a pharmaceutically acceptable salt thereof, or a solvate of a pharmaceutically acceptable salt thereof.

[0011] in,

[0012] L is -(CR) a R b ) n -, R a and R b Independently hydrogen (H) or deuterium (D), n is 2, 3, 4, 5 or 6;

[0013] R 1 Hydrogen, deuterium or

[0014] Y 1 Y 6 Y 7 Y 8 Y 9 Y 10 Y 11 Y 12 Y 13 Y 14 Y 15 Y 16 Y 17 Y 18 Y 19 Y 20 Y 21 Y 22 Y 23 Y 24 Y 25 Y 26 Y 27 Y 28 Y 29 Y 30 Y 31 Y 32Y 33 and Y 34 Independently hydrogen or deuterium;

[0015] And the L and R 1 Y 1 Y 6 Y 7 Y 8 Y 9 Y 10 Y 11 Y 12 Y 13 Y 14 Y 15 Y 16 Y 17 Y 18 Y 19 Y 20 Y 21 Y 22 Y 23 Y 24 Y 25 Y 26 Y 27 Y 28 Y 29 Y 30 Y 31 Y 32 Y 33 and Y 34 At least one of them contains deuterium.

[0016] In one embodiment, certain groups in the compound shown in formula (I) have the following definitions, and the definitions of groups not mentioned are as described in any embodiment of the present invention (hereinafter referred to as "in one embodiment").

[0017] In one of the schemes, Y 1 Y 9 Y 10 Y 15 Y 16 Y 17 Y 18 Y 19 Y 20 Y 21 Y 22 Y 23 Y 24 Y 25 Y 26 and Y 34 It is hydrogen.

[0018] In one particular scheme, L is n1 is 0, 1, 2, 3, or 4, with the "1" position connected to an oxygen atom; Y 2Y 3 Y 4 and Y 5 Independently hydrogen or deuterium; preferably, L is For example, L is

[0019] In one particular scheme, R 1 for For example, methyl or In one of the solutions, for

[0020] In one of the solutions, for In one of the solutions, for

[0021] In one embodiment, the pharmaceutically acceptable salt is a formate, such as one molecule of formate.

[0022] In this application, the deuterium isotope content at the deuterium substitution site is greater than the natural deuterium isotope content (about 0.015%). In one embodiment, it is preferably more than 30%, more preferably more than 50%, more preferably more than 75%, more preferably more than 95%, more preferably more than 96%, more preferably more than 97%, more preferably more than 98%, and more preferably more than 99%.

[0023] In one embodiment, the compound shown in formula (I) is the compound shown in formula I-1.

[0024] n1 is 1, 2, 3, or 4, R 1 Y 9 Y 10 Y 11 Y 12 Y 13 Y 14 Y 15 Y 16 Y 17 Y 18 Y 19 Y 21 Y 22 Y 23 Y 24 Y 25 Y 26 Y 27 Y 28 Y 29 Y 30 Y 31 Y 32Y 33 and Y 34 The definition is as described in the previous scheme.

[0025] In one embodiment, the compound shown in formula (I) is the compound shown in formula I-2.

[0026] L, Y 9 Y 10 Y 11 Y 12 Y 13 Y 14 Y 15 Y 16 Y 17 Y 18 Y 19 Y 21 Y 22 Y 23 Y 24 Y 25 Y 26 Y 27 Y 28 Y 29 Y 30 Y 31 Y 32 Y 33 and Y 34 The definition is as described in the previous scheme.

[0027] In one embodiment, the compound shown in formula (I) is a compound shown in formula I-3.

[0028] L, R 1 Y 11 Y 12 Y 13 Y 14 Y 17 Y 18 Y 19 Y 21 Y 22 Y 23 Y 24 Y 25 Y 26 Y 27 Y 28 Y 29 Y 30 Y 31 Y 32 Y 33 and Y 34 The definition is as described in the previous scheme.

[0029] In one embodiment, the compound shown in formula (I) is a compound shown in formula I-4.

[0030] L, R 1 Y 9 Y 10 Y 13 Y 14 Y 15 Y 16 Y 17 Y 18 Y 19 Y 21 Y 22 Y 23 Y 24 Y 25 Y 26 Y 27 Y 28 Y 29 Y 30 Y 31 Y 32 Y 33 and Y 34 The definition is as described in the previous scheme;

[0031] Preferably, the compound shown in formula (I) is the compound shown in formula I-4-1.

[0032] In one embodiment, the compound shown in formula (I) is a compound shown in formulas I-5.

[0033] L, R 1 Y 9 Y 10 Y 11 Y 12 Y 13 Y 14 Y 15 Y 16 Y 17 Y 18 Y 19 Y 21 Y 22 Y 23 Y 24 Y 25 Y 26 Y 27 Y 28 Y 29 Y 30 Y 31 Y 32 and Y 33 The definition is as described in the previous scheme.

[0034] In one embodiment, the compound shown in formula (I) is a compound shown in formulas I-6.

[0035] L, R 1 Y 9 Y 10 Y 11 Y 12 Y 13 Y 14 Y 15 Y 16 Y 17 Y 18 Y 19 Y 21 Y 22 Y 23 Y 24 Y 25 Y 26 Y 34 and Y 33 The definition is as described in the previous scheme.

[0036] In one embodiment, the compound shown in formula (I) is a compound shown in formulas I-7.

[0037] L, R 1 Y 9 Y 10 Y 11 Y 12 Y 13 Y 14 Y 15 Y 16 Y 17 Y 18 Y 19 Y 21 Y 22 Y 23 Y 24 Y 25 Y 26 Y 27 Y 28 Y 29 Y 30 Y 31 Y 32 and Y 34 The definition is as described in the previous scheme.

[0038] In one embodiment, the compound shown in formula (I) is a compound shown in formula I-8.

[0039] L, R1 Y 9 Y 10 Y 11 Y 12 Y 13 Y 14 Y 15 Y 16 Y 17 Y 18 Y 19 Y 21 Y 22 Y 23 Y 24 Y 27 Y 28 Y 29 Y 30 Y 31 Y 32 Y 33 and Y 34 The definition is as described in the previous scheme.

[0040] In one embodiment, the compound shown in formula (I) is a compound shown in formula I-9.

[0041] L, R 1 Y 9 Y 10 Y 11 Y 12 Y 13 Y 14 Y 15 Y 16 Y 17 Y 18 Y 19 Y 21 Y 25 Y 26 Y 27 Y 28 Y 29 Y 30 Y 31 Y 32 Y 33 and Y 34 The definition is as described in the previous scheme.

[0042] In one embodiment, the compound shown in formula (I) is the compound shown in formula I-10.

[0043] L, R 1 Y 9 Y 10 Y 11 Y 12 Y13 Y 14 Y 15 Y 16 Y 18 Y 19 Y 21 Y 22 Y 23 Y 24 Y 25 Y 26 Y 27 Y 28 Y 29 Y 30 Y 31 Y 32 Y 33 and Y 34 The definition is as described in the previous scheme.

[0044] In one embodiment, the compound shown in formula (I) is the compound shown in formula I-11.

[0045] L, R 1 Y 9 Y 10 Y 11 Y 12 Y 13 Y 14 Y 15 Y 16 Y 17 Y 19 Y 21 Y 22 Y 23 Y 24 Y 25 Y 26 Y 27 Y 28 Y 29 Y 30 Y 31 Y 32 Y 33 and Y 34 The definition is as described in the previous scheme.

[0046] In one embodiment, the compound shown in formula (I) is the compound shown in formula I-12.

[0047] L, R 1 Y 9 Y 10 Y 11 Y 12 Y 13 Y 14Y 15 Y 16 Y 17 Y 18 Y 21 Y 22 Y 23 Y 24 Y 25 Y 26 Y 27 Y 28 Y 29 Y 30 Y 31 Y 32 Y 33 and Y 34 The definition is as described in the previous scheme;

[0048] Preferably, the compound shown in formula (I-12) is the compound shown in formula I-12-1.

[0049] In one embodiment, the compound shown in formula (I) is the compound shown in formula I-13.

[0050] L, R 1 Y 9 Y 10 Y 11 Y 12 Y 13 Y 14 Y 15 Y 16 Y 17 Y 18 Y 19 Y 22 Y 23 Y 24 Y 25 Y 26 Y 27 Y 28 Y 29 Y 30 Y 31 Y 32 Y 33 and Y 34 The definition is as described in the previous scheme.

[0051] In one embodiment, the compound represented by formula (I) is any of the following compounds:

[0052] In one embodiment, the pharmaceutically acceptable salt of the compound represented by formula (I) is any of the following compounds:

[0053] Preferably, the formate is a single molecule of formate.

[0054] This invention provides a pharmaceutical composition comprising:

[0055] (1) (therapeuticly effective amount) of the compound of formula (I) as described in any embodiment of the invention, its solvates, pharmaceutically acceptable salts thereof, or solvates of pharmaceutically acceptable salts thereof; and

[0056] (2) Pharmaceutical excipients.

[0057] The present invention provides the use of a compound of formula (I) as described in any embodiment of the present invention, a solvate thereof, a pharmaceutically acceptable salt thereof, a solvate of a pharmaceutically acceptable salt thereof, or the above pharmaceutical composition in the preparation of JAK2 kinase and / or FLT3 kinase inhibitors.

[0058] The present invention provides the use of a compound of formula (I) as described in any embodiment of the present invention, a solvate thereof, a pharmaceutically acceptable salt thereof, a solvate of a pharmaceutically acceptable salt thereof, or the above-described pharmaceutical composition in the preparation of a medicament for treating and / or preventing diseases mediated by JAK2 kinase and / or FLT3 kinase.

[0059] In one embodiment, the disease mediated by JAK2 kinase and / or FLT3 kinase is a hematologic malignancy, such as acute myeloid leukemia, chronic myeloid leukemia, myeloma, acute lymphoblastic leukemia, acute myeloid leukemia, acute promyelocytic leukemia, chronic lymphocytic leukemia, chronic neutrophilic leukemia, acute undifferentiated cell leukemia, myelodysplastic syndrome, myelodysplastic syndrome, myelofibrosis, multiple myeloma, spinal sarcoma, or primary thrombocytosis.

[0060] The present invention provides the use of a compound of formula (I) as described in any embodiment of the present invention, a solvate thereof, a pharmaceutically acceptable salt thereof, a solvate of a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof in the preparation of a medicament for the treatment and / or prevention of hematologic malignancies.

[0061] In one embodiment, the hematologic malignancy is acute myeloid leukemia, chronic myeloid leukemia, myeloma, acute lymphoblastic leukemia, acute myeloid leukemia, acute promyelocytic leukemia, chronic lymphocytic leukemia, chronic neutrophilic leukemia, acute undifferentiated cell leukemia, myelodysplastic syndrome, myelodysplastic syndrome, myelofibrosis, multiple myeloma, spinal cord sarcoma, or primary thrombocytosis.

[0062] Unless otherwise specified, the terms used in this invention have the following meanings:

[0063] The term "pharmaceutically acceptable salt" refers to a salt obtained by reacting a compound with a pharmaceutically acceptable (relatively non-toxic, safe, and suitable for patient use) acid or base. When a compound contains a relatively acidic functional group, a base addition salt can be obtained by contacting the free form of the compound with a sufficient amount of a pharmaceutically acceptable base in a suitable inert solvent. When a compound contains a relatively basic functional group, an acid addition salt can be obtained by contacting the free form of the compound with a sufficient amount of a pharmaceutically acceptable acid in a suitable inert solvent.

[0064] The term "pharmaceutically acceptable" means that, within a reasonable medical judgment, it is suitable for contact with human and other mammalian tissues without undue toxicity, irritation, allergic reactions, etc., and that when administered to a recipient, it can directly or indirectly provide the compound or prodrug of the present invention.

[0065] The term "solvate" refers to a substance formed by the crystallization of a compound with a solvent (including but not limited to water, methanol, ethanol, etc.). Solvates are classified into stoichiometric and non-stoichiometric solvates. "Hydrate" refers to a compound that further binds stoichiometric or non-stoichiometric water through non-covalent intermolecular forces.

[0066] The term "therapeutic effective amount" refers to the amount of compound administered to a patient that is sufficient to effectively treat the disease. Therapeutic effective amount will vary depending on the compound, the type of disease, the severity of the disease, the patient's age, etc., but may be adjusted as appropriate by those skilled in the art.

[0067] The term "pharmaceutical excipients" refers to the excipients and additives used in the production of pharmaceuticals and the preparation of prescriptions. They are all substances contained in pharmaceutical preparations, excluding the active ingredient.

[0068] The term “treatment” refers to any of the following: (1) alleviating one or more biological manifestations of a disease; (2) interfering with one or more points in the biological cascade that triggers the disease; or (3) slowing the development of one or more biological manifestations of a disease.

[0069] The term "prevention" refers to reducing the risk of developing a disease.

[0070] The term "patient" refers to any animal that has received or is about to receive treatment, preferably a mammal, and most preferably a human. Mammals include, but are not limited to, cattle, horses, sheep, pigs, cats, dogs, mice, rats, rabbits, guinea pigs, monkeys, and humans.

[0071] Based on common knowledge in the field, the above-mentioned preferred conditions can be combined arbitrarily to obtain various preferred embodiments of the present invention.

[0072] The reagents and raw materials used in this invention are all commercially available.

[0073] The compounds provided by this invention can serve as kinase inhibitors with dual targets of JAK2 and FLT3, or as kinase inhibitors with single targets of JAK2 or FLT3, offering new options for the preparation of drugs for the treatment and / or prevention of tumors. Compared with known non-deuterated compounds in the prior art, the main advantages are:

[0074] (1) By using deuteration, the metabolism of compounds in organisms can be altered, resulting in compounds with better pharmacokinetic parameters. In this case, dosage can be changed and long-acting formulations can be formed, improving applicability.

[0075] (2) Replacing hydrogen atoms in a compound with deuterium can increase the drug concentration in animals due to its deuterium isotope effect, thereby improving the drug's efficacy.

[0076] (3) Replacing hydrogen atoms in a compound with deuterium may improve the safety of the compound because some metabolites are inhibited. Detailed Implementation

[0077] The present invention is further illustrated below by way of embodiments, but the invention is not limited to the scope of the embodiments described herein. Experimental methods in the following embodiments that do not specify specific conditions were performed according to conventional methods and conditions, or as selected according to the product instructions.

[0078] Example 1: Preparation of 2-((1-(2-fluoro-4-)((4-(1-isopropyl-1H-pyrazol-4-yl)-5-methylpyrimidin-2-yl)amino)phenyl)piperidin-4-yl)(methyl-D3)amino)ethane-1-ol (compound D001)

[0079] 1. Preparation of 1-(2-fluoro-4-nitrophenyl)-N-(methyl-d3)piperidin-4-amine (Compound 2)

[0080] A solution of compound 1 (10.0 g, 42.0 mmol), trideuterium methylamine hydrochloride (4.3 g, 62.9 mmol), glacial acetic acid (5.0 g, 83.9 mmol, 4.8 mL), and sodium borohydride acetate (17.8 g, 83.9 mmol) were added to dichloromethane (200 mL), and the mixture was stirred at 25 °C for 2 hours. The reaction mixture was diluted with 100 mL of sodium bicarbonate solution and then extracted with 300 mL of dichloromethane. The combined organic layer was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain a residue. The crude product was slurried with dichloromethane:petroleum ether = 1:10. Compound 2 (10 g, 39.02 mmol, 92.95% yield) was a yellow solid.

[0081] 1 ¹H NMR (400MHz, DMSO): δppm 7.92-8.06 (m, 2H), 7.10-7.21 (m, 1H), 3.66 (d, J = 12.8Hz, 2H), 2.96 (t, J = 11.2Hz, 2H), 2.67 (s, 1H), 1.96 (d, J = 12.0Hz, 2H), 1.39-1.51 (m, 2H). LCMS monitoring: MS (ESI) m / z = 257.2 [M+H] + .

[0082] 2. Preparation of 2-((1-(2-fluoro-4-nitrophenyl)piperidin-4-yl)(methyl-d3)amino)ethane-1-ol (compound 3)

[0083] Potassium carbonate (5.39 g, 39.0 mmol) and 2-bromoethanol (3.7 g, 29.3 mmol, 2.0 mL) were added to a solution of acetonitrile (100 mL) containing compound 2 (5.0 g, 19.5 mmol), and the mixture was stirred at 85 °C for 1 hour. The reaction mixture was diluted with 200 mL of water and then extracted with 600 mL of ethyl acetate. The bound organic layer was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain a residue. Purification by column chromatography (dichloromethane / anhydrous methanol = 500 / 1 to 5 / 1, dichloromethane / anhydrous methanol = 4 / 1) yielded compound 3 (1.00 g, 3.33 mmol, yield 17.7%) as a yellow solid. LCMS (ESI) m / z = 301.1 [M+H] +

[0084] 3. Preparation of 2-((1-(4-amino-2-fluorophenyl)piperidin-4-yl)(methyl-d3)amino)ethane-1-ol (compound 4)

[0085] Compound 3 (1 g, 3.33 mmol) was dissolved in methanol (30.0 mL), and wet Pd / C (354 mg, 333 μmol, 10% purity) was added. The mixture was stirred for 8 hours at 25 °C under a hydrogen atmosphere and a pressure of 15 psi. Thin-layer chromatography showed no residue of compound 3. The reaction mixture was concentrated under reduced pressure to remove the solvent, giving compound 4 (900 mg, 3.33 mmol, 99.9% yield) as a yellow solid.

[0086] 1 HNMR (400MHz, CDCl3): δppm 6.75(t,J=9.2Hz,1H),6.16-6.42(m,2H),4.93(s,2H),4.27(t,J=5.2Hz,1H),3.43(q,J=6.0Hz,2H),3.11(d,J=11.6Hz, 2H),2.54(s,2H),2.45-2.49(m,2H),2.36(tt,J=11.2,3.6Hz,1H),1.71(d,J=12.0Hz,2H),1.52(qd,J=12.0,3.6Hz,2H)

[0087] 4. Preparation of 2-((1-(2-fluoro-4-)((4-(1-isopropyl-1H-pyrazol-4-yl)-5-methylpyrimidin-2-yl)amino)phenyl)piperidin-4-yl)(methyl-d3)amino)ethane-1-ol (compound D001)

[0088] Compound 4 (131 mg, 555 μmol), compound 5 (150 mg, 555 μmol), Pd2(dba)3 (115 mg, 111 μmol), Xantphos (128 mg, 222 μmol), cesium carbonate (542 mg, 1.66 mmol), and 1,4-dioxane (5 mL) were added sequentially to a reaction flask. The mixture was purged three times with nitrogen, and the reaction was carried out at 100 °C under a nitrogen atmosphere for 2 hours. The reaction solution was concentrated under vacuum to obtain the crude product. The crude product was purified by column chromatography (dichloromethane / anhydrous methanol = 100 / 1 to 7 / 1, dichloromethane / anhydrous methanol = 5 / 1). The crude product was purified by preparative high-performance liquid chromatography (mobile phase: formic acid system) and lyophilized. The compound D001 formate (salt ratio 1:1) (63.2 mg, 122 μmol, yield 22.1%, purity 100%, deuteration rate 95%) was obtained as a white solid.

[0089] Preparative separation conditions for high performance liquid chromatography (HPLC): Instrument model: Gilson 281+shamaduzu LC20AP Semi-preparative HPLC system; Column: CD04-Welch Ultimate C18 150*25mm*7um; Mobile phase: A is water (0.225% formic acid), B is acetonitrile; Gradient: Initial gradient of mobile phase A 12%, final gradient of mobile phase A 42%, gradient time 10min; Flow rate: 25mL / min;

[0090] 1 H NMR(400MHz,CHLOROFORM-d)δppm 8.44(s,1H),8.18(s,1H),8.09(d,J=12.0Hz,2H),7.76(dd,J=14.8,2.0Hz,1H),7.37(s,1H),7.14(d d,J=8.4,2.0Hz,1H),6.92(t,J=9.2Hz,1H),4.59(dt,J=13.2,6.4Hz,1H),3.95-3.99(m,2H),3.52(d ,J=11.6Hz,2H), 3.26(tt,J=12.0,3.2Hz,1H), 3.05-3.14(m,2H), 2.78(t,J=11.2Hz,2H), 2.38(s,3H), 2.16(d,J=11.2Hz,2H), 1.89-2.05(m,2H), 1.60(d,J=6.8Hz,6H). Monitoring LCMS: MS(ESI) m / z=471.5[M+H] +

[0091] Example 2: Preparation of 2-((1-(2-fluoro-4-)((4-(1-isopropyl-1H-pyrazol-4-yl)-5-methylpyrimidin-2-yl)amino)phenyl)piperidin-4-yl)(methyl)amino)ethyl-1,1,2,2-d4-1-ol (compound D002)

[0092] 1. Preparation of 1-(2-fluoro-4-nitrophenyl)-N-methylpiperidin-4-amine (Compound 2)

[0093] A solution of compound 1 (10.0 g, 42.0 mmol), methylamine hydrochloride (4.25 g, 62.9 mmol), glacial acetic acid (5.0 g, 83.9 mmol, 4.8 mL), and sodium borohydride acetate (17.8 g, 83.9 mmol) were added to dichloromethane (200 mL), and the mixture was stirred at 25 °C for 2 hours. The reaction mixture was diluted with 100 mL of sodium bicarbonate solution and then extracted with 300 mL of dichloromethane (100 mL * 3). The combined organic layer was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain the residue. The crude product was pulped with dichloromethane: petroleum ether = 1:10. Compound 2 (9 g, 35.53 mmol, 84.65% yield) was a yellow solid.

[0094] 1 ¹H NMR (EC29003-4-p1a1, 400MHz, DMSO): δppm 7.88-8.18 (m, 2H), 7.15 (t, J = 9.2Hz, 1H), 3.64 (d, J = 12.8Hz, 2H), 2.98 (t, J = 11.2Hz, 2H), 2.53-2.63 (m, 1H), 2.30 (s, 3H), 1.92 (d, J = 11.2Hz, 2H), 1.28-1.47 (m, 2H). LCMS monitoring showed MS (ESI) m / z = 254.2 [M+H]. +

[0095] 2. Preparation of 2-((1-(2-fluoro-4-nitrophenyl)piperidin-4-yl)(methyl)amino)ethane-1,1,2,2-d4-1-ol (Compound 3)

[0096] Potassium carbonate (1.09 g, 7.90 mmol) and 2-bromo-1,1,2,2-tetradeuterylethanol (764 mg, 5.92 mmol) were added to a solution of acetonitrile (20 mL) containing compound 2 (1.0 g, 3.95 mmol), and stirred at 85 °C for 1 hour. The reaction mixture was diluted with 20 mL of water and then extracted with 60 mL of ethyl acetate (20 mL * 3). The bound organic layer was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain the residue. Purification by column chromatography (dichloromethane / anhydrous methanol = 500 / 1 ~ 5 / 1, dichloromethane / anhydrous methanol = 4 / 1) gave compound 3 (500 mg, 1.66 mmol, yield 42.0%) as a yellow solid. LCMS monitoring: MS (ESI) m / z = 302.0 [M + H] +

[0097] 3. Preparation of 2-((1-(4-amino-2-fluorophenyl)piperidin-4-yl)(methyl)amino)ethane-1,1,2,2-d4-1-ol (compound 4)

[0098] Compound 3 (200 mg, 664 mmol) was dissolved in methanol (10.0 mL), and wet Pd / C (70.63 mg, 66.3 μmol, 10% purity) was added. The mixture was stirred for 8 hours at 25 °C under a hydrogen atmosphere and a pressure of 15 psi. The reaction mixture was concentrated under reduced pressure to remove the solvent, yielding compound 4 (200 mg, 737 μmol, crude) as a yellow solid.

[0099] 4. Preparation of 2-((1-(2-fluoro-4-)((4-(1-isopropyl-1H-pyrazol-4-yl)-5-methylpyrimidin-2-yl)amino)phenyl)piperidin-4-yl)(methyl)amino)ethyl-1,1,2,2-d4-1-ol (compound D002)

[0100] Compound 4 (150 mg, 552 μmol), compound 5 (131 mg, 553 μmol), Pd₂(dba)₃.CHCl₃ (31.3 mg, 55.3 μmol), Xantphos (63.97 mg, 111 μmol), cesium carbonate (540 mg, 1.66 mmol), and 1,4-dioxane (5 mL) were added sequentially to a reaction flask. The mixture was purged three times with nitrogen, and the reaction was carried out at 100 °C under a nitrogen atmosphere for 2 hours. The reaction solution was concentrated under vacuum to obtain a crude product. The crude product was purified by column chromatography (dichloromethane / anhydrous methanol = 100 / 1 to 7 / 1, dichloromethane / anhydrous methanol = 5 / 1). The crude product was then purified by adding acetonitrile (4 mL) at 70 °C. The compound D002 (25.0 mg, 52.3 μmol, yield 9.41%, purity 98.711%, deuteration rate 95%) was obtained as a white solid.

[0101] 1 H NMR(400MHz,CHLOROFORM-d)δppm 8.12(s,1H),8.00(d,J=3.2Hz,2H),7.64(dd,J=14.6,2.4Hz,1H),7.03(dd,J=8. 4,1.6Hz,1H),6.78-6.90(m,2H),4.51(dt,J=13.2,6.8Hz,1H),3.39(d,J=12.0H z,2H),2.60(td,J=11.2,3.2Hz,2H),2.49(dd,J=6.0,3.2Hz,1H),2.23-2.31(m, 6H), 1.72-1.84 (m, 4H), 1.59 (d, J=6.8Hz, 6H). Monitoring LCMS: MS (ESI) m / z=472.3[M+H] +

[0102] 5. Preparation of 2-((1-(2-fluoro-4-)((4-(1-isopropyl-1H-pyrazol-4-yl)-5-methylpyrimidin-2-yl)amino)phenyl)piperidin-4-yl)(methyl)amino)ethyl-1,1,2,2-d4-1-ol formate (compound D002 formate)

[0103] D002 (23 mg, 48.77 μmol) and formic acid (4.69 mg, 97.54 μmol) were dissolved in water (1 mL). The mixture was stirred at 25 °C for 0.5 hours under a nitrogen atmosphere and then freeze-dried. D002 formate (salt ratio 1:1) (23.71 mg, 45.80 μmol, yield 93.92%, purity 100%) was obtained as a white solid.

[0104] 1 H NMR(400MHz,CHLOROFORM-d)δppm 8.45(s,1H),8.19(s,1H),8.10(s,1H),8.07(s,1H),7.74(dd,J=14.4,2 .0Hz,1H),7.10–7.14(m,2H),6.92(t,J=8.8Hz,2H),4.59(dt,J=13.2,6. 8Hz,1H),3.50(d,J=12.0Hz,2H),3.18–3.21(m,1H),2.74-2.80(m,2H),2 .37(s,3H),2.15-2.18(m,2H),1.93-1.99(m,2H),1.59(d,J=6.4Hz,6H).

[0105] Example 3: Preparation of 2-((1-(2-fluoro-4-)((4-(1-isopropyl-1H-pyrazol-4-yl)-5-methylpyrimidin-2-yl)amino)phenyl)piperidin-4-yl-2,2,6,6-d4)(methyl)amino)ethane-1-ol (compound D004)

[0106] 1. Preparation of 1-benzylpiperidine-2,2,6,6-d4-4-ol (compound 2)

[0107] Compound 1 (10 g, 62.43 mmol) was added to benzylamine (5.52 g, 24.97 mmol) and sonicated for 10 min, then stirred at 20 °C for 1 h. Propylene (trimethyl)silane (3.42 g, 29.96 mmol) was added to the solution, and the mixture was heated at 40 °C for 16 h. The reaction mixture was diluted with 100 mL of potassium carbonate solution, stirred for 10 min, and then extracted with 300 mL of dichloromethane. The combined organic layer was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain the residue. The residue was purified by preparative chromatography (dichloromethane:methanol = 5:1). Compound 2 (2.5 g, 12.8 mmol, 51.2% yield) was a colorless oil.

[0108] 1 HNMR (400MHz, CDCl3): ppm 7.25-7.34(m,5H),3.70(dq,J=8.8,4.4Hz,1H),3.51(s,2H),1.87(dd,J=12.8,4.0Hz,2H)1.54-1.64(m,2H).

[0109] Monitoring LCMS: MS(ESI) m / z = 196.2 [M+H] +

[0110] 2. Preparation of piperidine-2,2,6,6-d4-4-ol (compound 3)

[0111] Pd(OH)₂ / C (863 mg, 1.23 mmol, 20% purity) was added to a methanol (50.0 mL) solution containing compound 2 (2.40 g, 12.3 mmol), and stirred at 25 °C and 50 PSI for 8 hours. The reaction mixture was filtered and concentrated under reduced pressure to give crude product compound 3 as a colorless oil.

[0112] 3. Preparation of 1-(2-fluoro-4-nitrophenyl)piperidin-4-one-2,2,6,6-d4 (compound 4)

[0113] Compound 3a (2.27 g, 14.3 mmol) was dissolved in methanol (30.0 mL), and triethylamine (2.89 g, 28.5 mmol) and compound 3 (1.50 g, 14.3 mmol) were added. The mixture was stirred at 50 °C for 2 hours. The reaction mixture was concentrated under reduced pressure to remove the solvent, giving a residue. The residue was purified by preparative chromatography (commercial hexane: ethyl acetate = 100 / 1 to 1 / 1). Compound 4 (2.00 g, 8.19 mmol, yield 57.4%) was given as a yellow solid.

[0114] 4. Preparation of 2-((1-(2-fluoro-4-nitrophenyl)piperidin-4-yl-2,2,6,6-d4)(methyl)amino)ethane-1-ol (compound 5)

[0115] Compound 4 (700 mg, 2.89 mmol), 2-(methylamine)ethanol (326 mg, 4.33 mmol), NaBH(OAc)3 (1.22 g, 5.78 mmol), and glacial acetic acid (347 mg, 5.78 mmol) were added to a solution of dichloromethane (10 mL), and the mixture was stirred at 25 °C for 2 hours. The reaction mixture was concentrated under reduced pressure to remove the solvent. The solution was diluted with 40 mL of sodium carbonate solution and extracted with 120 mL of dichloromethane (40 mL * 3). The combined organic layers were dried over sodium sulfate, filtered, and concentrated under reduced pressure to obtain the residue. The residue was purified by column chromatography (dichloromethane / anhydrous methanol = 100 / 1 to 10 / 1, dichloromethane / anhydrous methanol = 5 / 1). Compound 5 (700 mg, 2.32 mmol, yield 80.4%) was given as a yellow solid.

[0116] Monitoring LCMS: MS(ESI) m / z = 302.2 [M+H] + .

[0117] 5. Preparation of 2-((1-(4-amino-2-fluorophenyl)piperidin-4-yl-2,2,6,6-d4)(methyl)amino)acetic acid-1-ol (compound 6)

[0118] Pd / C (247 mg, 10% purity) was dissolved in methanol (10.0 mL), and compound 5 (0.7 g, 2.32 mmol) was added under a nitrogen atmosphere. The suspension was degassed and purged with hydrogen three times. The mixture was stirred at 25 °C for 3 hours under hydrogen (50 Psi). The reaction mixture was filtered and concentrated under reduced pressure to obtain a residue. Compound 6 (500 mg, 1.84 mmol, 79.32% yield) was given as a brown solid and could be used directly in the next step without purification.

[0119] 1 H NMR (400MHz, DMSO-d6): δppm 7.37(t,J=9.2Hz,1H),6.87-6.98(m,2H),4.07(t,J=6.4Hz,2H),3.13-3.16(m,2H),3.0 3(tt,J=11.2,3.2Hz,1H),2.85(s,3H),2.34(dd,J=12.4,3.2Hz,2H),2.10-2.20(m,2H).

[0120] 6. Preparation of 2-((1-(2-fluoro-4-)((4-(1-isopropyl-1H-pyrazol-4-yl)-5-methylpyrimidin-2-yl)amino)phenyl)piperidin-4-yl-2,2,6,6-d4)(methyl)amino)ethane-1-ol (compound D004)

[0121] Compound 6 (250 mg, 921 μmol), compound 6a (218 mg, 921 μmol), Pd2(dba)3 (52.19 mg, 92.13 μmol), Xantphos (107 mg, 184 μmol), cesium carbonate (900 mg, 2.76 mmol), and 1,4-dioxane (5 mL) were added sequentially to a reaction flask. The mixture was purged three times with nitrogen, and the reaction was carried out at 100 °C under a nitrogen atmosphere for 2 hours. The reaction solution was concentrated under vacuum to obtain a crude product. The crude product was purified by column chromatography (dichloromethane / anhydrous methanol = 100 / 1 to 7 / 1, dichloromethane / anhydrous methanol = 5 / 1). The crude product was then purified by adding acetonitrile (4 mL) at 70 °C. Compound D004 (38.4 mg, 80.5 μmol, yield 8.74%, purity 98.9%, deuteration rate 95%) was obtained as a white solid.

[0122] 1 H NMR(400MHz,CHLOROFORM-d)δppm 8.20(s,1H),8.08(d,J=3.13Hz,2H),7.71(dd,J=14.8,24Hz,1H),7.06-7.14(m,1H),7.0 0(s,1H),6.89-6.96(m,1H),4.58(dt,J=13.2,6.4Hz,1H),3.59(t,J=5.2Hz,2H),3.01(br s,1H),2.67(t,J=5.2Hz,2H),2.47-2.60(m,1H),2.37(s,3H),2.32(s,3H),1.78-1.85(m,4H),1.59(d,J=6.4Hz,6H).

[0123] Monitoring LCMS: MS(ESI) m / z = 472.5 [M+H] + .

[0124] 7. Preparation of 2-((1-(2-fluoro-4-)((4-(1-isopropyl-1H-pyrazol-4-yl)-5-methylpyrimidin-2-yl)amino)phenyl)piperidin-4-yl-2,2,6,6-d4)(methyl)amino)ethane-1-ol formate (compound D004 formate)

[0125] D004 (37 mg, 78.46 μmol) and formic acid (7.54 mg, 156.91 μmol) were dissolved in water (1 mL). The mixture was stirred at 25 °C for 0.5 hours under a nitrogen atmosphere and then freeze-dried. D004 formate (salt ratio 1:1) (40 mg, 77.28 μmol, yield 98.50%) was obtained as a white solid.

[0126] 1 H NMR(400MHz,CHLOROFORM-d)δppm 8.50(s,1H),8.19(s,1H),8.10(s,1H),8.07(s,1H),7.74(dd,J=14.8,24Hz,1 H),7.06-7.14(m,2H),6.91(t,J=8.8,1H),4.59(dt,J=13.2,6.4Hz,1H),3.90( t,J=4.8Hz,2H),3.05-3.14(m,2H),3.02(t,J=4.8Hz,2H),2.71(s,3H),2.37(s ,3H),2.10(dd,J=3.6,12.4Hz,2H),1.96(t,J=12.0,2H),1.59(d,J=6.4Hz,6H)

[0127] Example 4: Preparation of 2-((1-(2-fluoro-4-((5-methyl-4-)(1-(propyl-2-yl-1,1,1,3,3-d6)-1H-pyrazol-4-yl)pyrimidin-2-yl)amino)phenyl)piperidin-4-yl)(methyl)amino)ethane-1-ol (compound D008)

[0128] 1. Preparation of propan-1,1,1,3,3,3-d6-2-ol (compound 2)

[0129] A solution of lithium aluminum hydride (2.5 M, 13.41 mL) was added to tetrahydrofuran (25.0 mL). Compound 1 (5.00 g, 77.9 mmol, 5.73 mL) was added dropwise under a nitrogen atmosphere at 0 °C. The mixture was stirred at 0 °C for 1 hour to obtain a crude tetrahydrofuran solution of compound 2 (5.00 g, 75.6 mmol).

[0130] 2. Preparation of 1-(propyl-2-yl-1,1,1,3,3,3-d6)-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborhecyclopentan-2-yl)-1H-pyrazole (compound 3)

[0131] Triphenylphosphine (11.4 g, 43.3 mmol) and compound 2A (7.00 g, 36.1 mmol) were added to a tetrahydrofuran (100 mL) solution containing compound 2 (2.39 g, 36.1 mmol). DIAD (8.80 g, 43.3 mmol, 8.4 mL) was added dropwise at 0 °C, and the mixture was stirred for 2 hours. The reaction mixture was concentrated under reduced pressure to obtain a residue. Purification by column chromatography (petroleum ether / ethyl acetate = 500 / 1 to 5 / 1, petroleum ether / ethyl acetate = 3 / 1) gave compound 3 (700 mg, 2.89 mmol, yield 8.00%) as a colorless oily liquid.

[0132] 1 HNMR (400MHz, CDCl3): δppm 7.80(s,1H),7.74(s,1H),4.50(s,1H),1.32(s,12H).

[0133] Monitoring LCMS: MS(ESI) m / z = 243.1 [M+H] +

[0134] 3. Preparation of 2-chloro-5-methyl-4-(1-(propyl-2-yl-1,1,1,3,3,3-d6)-1H-pyrazol-4-yl)pyrimidine (compound 4)

[0135] Compound 3 (700 mg, 2.89 mmol), compound 3A (518 mg, 3.18 mmol), Pd(dppf)Cl2 (106 mg, 145 μmol), and potassium carbonate (1.20 g, 8.67 mmol) were dissolved in water (2.0 mL) and dioxane (7.0 mL), and stirred at 100 °C under a nitrogen atmosphere for 4 hours. The reaction mixture was concentrated under reduced pressure to remove the solvent, yielding a residue. Purification by column chromatography (petroleum ether / ethyl acetate = 10 / 1–3 / 1, petroleum ether / ethyl acetate = 5 / 1) gave compound 4 (500 mg, 1.94 mmol, yield 67.2%) as a white solid. LCMS (ESI) m / z = 243.3 [M+H] +

[0136] 4. Preparation of 2-((1-(2-fluoro-4-((5-methyl-4-)(1-(propyl-2-yl-1,1,1,3,3-d6)-1H-pyrazol-4-yl)pyrimidin-2-yl)amino)phenyl)piperidin-4-yl)(methyl)amino)ethane-1-ol (compound D008)

[0137] Compound 4 (120 mg, 494 μmol), compound 4A (132 mg, 494 μmol), and Pd2(dba)3 were used. .CHCl3 (51.2 mg, 49.4 μmol), Xantphos (57.2 mg, 98.9 μmol), cesium carbonate (483 mg, 1.48 mmol), and 1,4-dioxane (5 mL) were added sequentially to the reaction flask. The mixture was purged three times with nitrogen, and the reaction was carried out at 100 °C under a nitrogen atmosphere for 2 hours. The reaction solution was concentrated under vacuum to obtain the crude product. The crude product was purified by column chromatography (dichloromethane / anhydrous methanol = 100 / 1 to 7 / 1, dichloromethane / anhydrous methanol = 5 / 1). The crude product was further purified by high-performance liquid chromatography. Compound D008 (40.9 mg, 86.4 μmol, yield 17.5%, purity 100%, deuteration rate 97%) was obtained as a white solid.

[0138] Preparative separation conditions for high performance liquid chromatography:

[0139] Instrument model: Gilson 281+shamaduzu LC20AP Semi-preparative HPLC system

[0140] Column: CD04-Welch Ultimate C18 150*25mm*7um

[0141] Mobile phase: A is water (0.225% formic acid), B is acetonitrile; Gradient: initial gradient of mobile phase A is 12%, final gradient of mobile phase A is 42%, gradient time is 10 min; Flow rate: 25 mL / min.

[0142] 1 H NMR(400MHz,CHLOROFORM-d)δppm 8.44(s,1H),8.18(s,1H),8.09(d,J=12.0Hz,2H),7.76(dd,J=14.8,2.0Hz,1H),7.37(s,1H),7.14(d d,J=8.4,2.0Hz,1H),6.92(t,J=9.2Hz,1H),4.59(dt,J=13.2,6.4Hz,1H),3.95-3.99(m,2H),3.52(d ,J=11.6Hz,2H), 3.26(tt,J=12.0,3.2Hz,1H), 3.05-3.14(m,2H), 2.78(t,J=11.2Hz,2H), 2.38(s,3H), 2.16(d,J=11.2Hz,2H), 1.89-2.05(m,2H), 1.60(d,J=6.8Hz,6H). Monitoring LCMS: MS(ESI) m / z=474.5[M+H] +

[0143] 5. Preparation of 2-((1-(2-fluoro-4-((5-methyl-4-)(1-(propyl-2-yl-1,1,1,3,3-d6)-1H-pyrazol-4-yl)pyrimidin-2-yl)amino)phenyl)piperidin-4-yl)(methyl)amino)ethane-1-ol formate (compound D008 formate)

[0144] D008 (41 mg, 86.57 μmol) and formic acid (8.32 mg, 173.14 μmol) were dissolved in water (1 mL). The mixture was stirred at 25 °C for 0.5 hours under a nitrogen atmosphere and then freeze-dried. D008 formate (salt ratio 1:1) (44 mg, 84.67 μmol, 97.81% yield) was obtained as a white solid.

[0145] 1 H NMR(400MHz,CHLOROFORM-d)δppm 8.53(s,1H),8.20(s,1H),8.10(s,1H),8.07(s,1H),7.74(dd,J=14.8,2.4Hz ,1H),7.10(dd,J=8.8,2.0Hz,1H),6.98(s,1H),6.95(t,J=8.8,1H),4.55(s,1 H),3.81(t,J=4.8Hz,2H),3.50(d,J=11.6,1H),2.90-2.95(m,3H),2.73(t,J= 10.8Hz,4H),2.60(s,3H),2.37(s,3H),2.03-2.06(m,2H),1.90-1.95(m,2H).

[0146] Example 5: Preparation of 2-((1-(2-fluoro-4-((5-methyl-4-(1-(prop-2-yl-d7)-1H-pyrazol-4-yl)pyrimidin-2-yl)amino)phenyl)piperidin-4-yl)(methyl)amino)ethane-1-ol (compound D009)

[0147] 1. Preparation of 2-chloro-5-methyl-4-(1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazol-4-yl)pyrimidine (compound 2)

[0148] Compound 1 (4.50 g, 13.9 mmol), compound 1A (2.49 g, 15.3 mmol), Pd(dppf)Cl2 (508 mg, 694 μmol), and potassium carbonate (5.75 g, 41.63 mmol, 3 eq) were added to dioxane (5 mL) and water (10 mL). The mixture was purged three times with nitrogen, and then stirred at 100 °C under a nitrogen atmosphere for 4 hours. The reaction mixture was cooled to room temperature and quenched with 50 mL of water, then diluted with 50 mL of ethyl acetate and extracted with 150 mL of ethyl acetate. The combined organic layer was washed with 50 mL of saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain the residue. Purification was performed by column chromatography (petroleum ether / ethyl acetate = 10 / 1 to 3 / 1, petroleum ether / ethyl acetate = 3 / 1). Compound 2 (3.00 g, 8.83 mmol, yield 63.6%, purity 95.6%) was given as a white solid.

[0149] 2. Preparation of 2-chloro-5-methyl-4-(1H-pyrazol-4-yl)pyrimidine (compound 3)

[0150] Compound 2 (3.00 g, 9.23 mmol) was dissolved in dichloromethane (15 mL), and trifluoroacetic acid (23.0 g, 202 mmol, 15 mL) was added. The mixture was stirred at 25 °C for 2 hours. The reaction mixture was adjusted to pH 8 with 1 M sodium hydroxide solution, diluted with 50 mL of water, and extracted with 250 mL of ethyl acetate. The bound organic layer was washed with 50 mL of saturated sodium chloride, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to give compound 3 (1.70 g, 7.79 mmol, yield 84.4%, purity 89.2%) as a white solid.

[0151] 3. Preparation of 2-chloro-5-methyl-4-(1-(propyl-2-yl-d7)-1H-pyrazol-4-yl)pyrimidine (compound 4)

[0152] Compound 3 (1.00 g, 5.14 mmol), cesium carbonate (5.02 g, 15.4 mmol), and 2-bromo-1,1,1,2,3,3,3-heptadenopropane (1.26 g, 10.3 mmol) were purged three times with nitrogen in acetonitrile (15 mL), and then stirred at 80 °C under a nitrogen atmosphere for 16 hrs. The reaction mixture was quenched with 50 mL of water, diluted with 50 mL of ethyl acetate, and extracted with 150 mL of ethyl acetate. The bound organic layer was washed with 50 mL of saturated sodium chloride, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain the residue. The residue was purified by column chromatography (petroleum ether / ethyl acetate = 10 / 1 to 3 / 1, petroleum ether / ethyl acetate = 3 / 1). Compound 4 (600 mg, 2.46 mmol, 47.9%) was given as a white solid.

[0153] Monitoring LCMS: MS(ESI) m / z = 244.0 [M+H] +

[0154] 4. Preparation of 2-((1-(2-fluoro-4-((5-methyl-4-(1-(prop-2-yl-d7)-1H-pyrazol-4-yl)pyrimidin-2-yl)amino)phenyl)piperidin-4-yl)(methyl)amino)ethane-1-ol (compound D009)

[0155] Compound 4 (120 mg, 492 μmol), compound 4A (131.62 mg, 492.32 μmol), Pd2(dba)3 (28.6 mg, 50.5 μmol), Xantphos (58.4 mg, 101 μmol), and cesium carbonate (481 mg, 1.48 mmol) were added to dioxane (5 mL), purged three times with nitrogen, and then stirred at 100 °C under a nitrogen atmosphere for 2 hours. The reaction solution was concentrated under vacuum to obtain a crude product. Purification was performed by column chromatography (dichloromethane / anhydrous methanol = 100 / 1 to 7 / 1, dichloromethane / anhydrous methanol = 5 / 1). The crude product was purified by 4 mL of acetonitrile at 70 °C. Compound D009 (50.8 mg, 105 μmol, yield 21.3%, purity 98.1%, deuteration rate 97%) was obtained as a white solid. 1 H NMR(400MHz,CHLOROFORM-d)δ:20(s,1H),8.08(d,J=4.0Hz,2H),7.71(dd,J=14.8,2.4Hz,1H),7.10(dd,J=8.4,1.6Hz,1H),6.89-6.98(m,2H),3 .59(t,J=5.2Hz,2H),3.42-3.52(m,2H),3.06(s,1H),2.63-2.76(m,4H) ,2.51-2.60(m,1H),2.37(s,3H),2.31-2.35(m,3H),1.80-1.88(m,4H).

[0156] Monitoring LCMS: MS(ESI) m / z = 475.4 [M+H] +

[0157] 5. Preparation of 2-((1-(2-fluoro-4-((5-methyl-4-(1-(prop-2-yl-d7)-1H-pyrazol-4-yl)pyrimidin-2-yl)amino)phenyl)piperidin-4-yl)(methyl)amino)ethane-1-ol formate (compound D009 formate)

[0158] D009 (50 mg, 105.35 μmol) and formic acid (10.12 mg, 210.69 μmol) were dissolved in water (1 mL). The mixture was stirred at 25 °C for 0.5 hours under a nitrogen atmosphere and then freeze-dried. D009 formate (salt ratio 1:1) (54 mg, 103.72 μmol, 98.45% yield) was obtained as a white solid.

[0159] 1 H NMR(400MHz,CHLOROFORM-d)δppm 8.53(s,1H),8.20(s,1H),8.10(s,1H),8.07(s,1H),7.75(dd,J=14.8,2.4 Hz,1H),7.10(dd,J=8.8,2.0Hz,1H),6.99(s,1H),6.95(t,J=8.8,1H),3.8 4(t,J=4.8Hz,1H),3.50(d,J=12.0,1H),2.93–3.00(m,1H),2.73(t,J=10. 8Hz,4H),2.61(s,3H),2.37(s,3H),2.00-2.10(m,2H),1.93-1.95(m,2H).

[0160] Example 6: Preparation of 2-((1-(2-fluoro-4-((5-methyl-4-(1-(prop-2-yl-d7)-1H-pyrazol-4-yl)pyrimidin-2-yl)amino)phenyl)piperidin-4-yl)(methyl-d3)amino)ethane-1,1,2,2-d4-1-ol (compound D023)

[0161] 1. Preparation of 2-((1-(2-fluoro-4-((5-methyl-4-(1-(prop-2-yl-d7)-1H-pyrazol-4-yl)pyrimidin-2-yl)amino)phenyl)piperidin-4-yl)(methyl-d3)amino)ethane-1,1,2,2-d4-1-ol (compound D023)

[0162] Compound 1 (80.0 mg, 328 μmol), compound 1A (117 mg, 427 μmol), Pd2(dba)3 (18.6 mg, 32.8 μmol), Xantphos (38.0 mg, 65.6 μmol), and cesium carbonate (321 mg, 984 μmol) were added to dioxane (5 mL), purged three times with nitrogen, and then stirred at 100 °C under a nitrogen atmosphere for 4 hours. The reaction solution was concentrated under vacuum to obtain the crude product. Purification was performed by column chromatography (dichloromethane / anhydrous methanol = 100 / 1 to 7 / 1, dichloromethane / anhydrous methanol = 5 / 1). The crude product was purified by 4 mL of acetonitrile at 70 °C. Compound D023 (24.7 mg, 50.36 μmol, yield 15.3%, purity 98.2%, deuteration rate 98%) was obtained as a white solid. 1 H NMR(400MHz,CHLOROFORM-d)δ:8.20(s,1H),8.08(d,J=4.4Hz,2H),7.71(dd,J=14.8,2.4Hz,1H),7.10(dd,J=8.8,1.6Hz,1H), 6.88-6.97(m,2H),3.47(d,J=11.2Hz,2H),3.04(s,1H),2.62-2.71(m,2H),2.50-2.60(m,1H),2.37(s,3H),1.79-1.86(m,4H).

[0163] Monitoring LCMS: MS(ESI) m / z = 482.5 [M+H] +

[0164] 2. Preparation of 2-((1-(2-fluoro-4-((5-methyl-4-(1-(prop-2-yl-d7)-1H-pyrazol-4-yl)pyrimidin-2-yl)amino)phenyl)piperidin-4-yl)(methyl-d3)amino)ethane-1,1,2,2-d4-1-ol formate (compound D023 formate)

[0165] DO23 (26 mg, 53.97 μmol) and formic acid (4.97 mg, 107.94 μmol) were dissolved in water (1 mL). The mixture was stirred at 25 °C for 0.5 hours under a nitrogen atmosphere and then freeze-dried. DO23 formate (salt ratio 1:1) (25 mg, 47.37 μmol, 87.77% yield) was obtained as a white solid.

[0166] 1H NMR(400MHz,CHLOROFORM-d)δppm 8.53(s,1H),8.20(s,1H),8.10(s,1H),8.07(s,1H),7.75(dd,J=14.8,2.4Hz,1H),7.10(dd,J=8.8,2.0Hz,1H),6.99(s,1H),6.95( t,J=8.8,1H),3.50(d,J=12.0,1H),2.93-3.00(m,1H),2.74(t,J=10.8Hz,2H),2.37(s,3H),2.00-2.10(m,2H),1.93-1.95(m,2H).

[0167] Example 7: Preparation of 2-((1-(2-fluoro-4-((5-methyl-4-(1-(propyl-2-yl-1,1,1,3,3,3-d6)-1H-pyrazol-4-yl)pyrimidin-2-yl)amino)phenyl)piperidin-4-yl)(methyl-d3)amino)ethane-1,1,2,2-d4-1-ol (Compound D027)

[0168] 1. Preparation of 2-((1-(2-fluoro-4-((5-methyl-4-(1-(propyl-2-yl-1,1,1,3,3,3-d6)-1H-pyrazol-4-yl)pyrimidin-2-yl)amino)phenyl)piperidin-4-yl)(methyl-d3)amino)ethane-1,1,2,2-d4-1-ol (compound D027)

[0169] Compound 1 (80.0 mg, 328 μmol), compound 1A (117 mg, 427 μmol), Pd2(dba)3 (18.6 mg, 32.8 μmol), Xantphos (38.0 mg, 65.6 μmol), and cesium carbonate (322 mg, 984 μmol) were added to dioxane (5 mL), purged three times with nitrogen, and then stirred at 100 °C under a nitrogen atmosphere for 4 hours. The reaction solution was concentrated under vacuum to obtain the crude product. Purification was performed by column chromatography (dichloromethane / anhydrous methanol = 100 / 1 to 7 / 1, dichloromethane / anhydrous methanol = 5 / 1). The crude product was purified by 4 mL of acetonitrile at 70 °C. Compound D027 (41.1 mg, 84.1 μmol, yield 25.5%, purity 98.3%, deuteration rate 98%) was obtained as a white solid.

[0170] 1H NMR(400MHz,CHLOROFORM-d)δ:8.20(s,1H),8.08(d,J=4.0Hz,2H),7.71(dd,J=14.8,2.4Hz,1H),7.10(dd,J=8.8,1.6Hz,1H),6.90-6 .98(m,2H),4.55(s,1H),3.46(d,J=11.2Hz,2H),3.03(s,1H),2.62-2.71(m,2H),2.50-2.59(m,1H),2.37(s,3H),1.79-1.86(m,4H).

[0171] Monitoring LCMS: MS(ESI) m / z = 481.6 [M+H] +

[0172] 2. Preparation of 2-((1-(2-fluoro-4-((5-methyl-4-(1-(prop-2-yl-1,1,1,3,3,3-d6)-1H-pyrazol-4-yl)pyrimidin-2-yl)amino)phenyl)piperidin-4-yl)(methyl-d3)amino)ethane-1,1,2,2-d4-1-ol formate (compound D027 formate)

[0173] DO27 (42 mg, 87.37 μmol) and formic acid (8.03 mg, 174.7 μmol) were dissolved in water (1 mL). The mixture was stirred at 25 °C for 0.5 hours under a nitrogen atmosphere and then freeze-dried. DO27 formate (salt ratio 1:1) (44 mg, 83.53 μmol, 95.60% yield) was obtained as a white solid.

[0174] 1 H NMR(400MHz,CHLOROFORM-d)δppm 8.54(s,1H),8.20(s,1H),8.10(s,1H),8.07(s,1H),7.75(dd,J=14.8,2.4Hz,1H),7.12(dd,J=8.8,2.0Hz,1H),7.00(s,1H),6.92(t,J=8 .8,1H),4.55(s,1H),3.50(d,J=12.0,2H),3.00–3.03(m,1H),2.74(t,J=10.8Hz,2H),2.37(s,3H),2.00-2.10(m,2H),1.90-1.95(m,2H).

[0175] Example 8: Preparation of 2-((1-(2-fluoro-4-((4-(1-isopropyl-1H-pyrazol-4-yl)-5-methylpyrimidin-2-yl)amino)phenyl)piperidin-4-yl)(methyl-d3)amino)ethane-1,1,2,2-d4-1-ol (Compound D010)

[0176] 1. Preparation of 2-((1-(2-fluoro-4-nitrophenyl)piperidin-4-yl)(methyl-d3)amino)ethane-1,1,2,2-d4-1-ol (Compound 2)

[0177] Potassium carbonate (2.91 g, 21.1 mmol) and compound 1A (2.00 g, 15.8 mmol) were added to a solution of acetonitrile (50.0 mL) containing compound 1 (2.70 g, 10.5 mmol), and the mixture was stirred at 85 °C for 1 hour. The reaction mixture was concentrated under reduced pressure to obtain a residue. Purification was performed by column chromatography (dichloromethane / anhydrous methanol = 500 / 1 to 5 / 1, dichloromethane / anhydrous methanol = 5 / 1) to give compound 2 (1.3 g, 4.3 mmol, 40.5% yield) as a yellow oily liquid.

[0178] 1 HNMR (EC29003-43-p1a1, 400MHz, DMSO): δppm 7.92-8.06(m,2H),7.10-7.21(m,1H),3.66(d,J=12.8Hz,2H),2.96(t,J=11.2Hz,2H),2.67(s,1H),1.96(d,J=12.0Hz,2H),1.39-1.51(m,2H).

[0179] Monitoring LCMS: MS(ESI) m / z = 305.0 [M+H] +

[0180] 2. Preparation of 2-((1-(4-amino-2-fluorophenyl)piperidin-4-yl)(methyl-d3)amino)ethane-1,1,2,2-d4-1-ol (compound 3)

[0181] Compound 2 (1 g, 3.29 mmol) was dissolved in methanol (20.0 mL), and wet Pd / C (349 mg, 328 μmol, 10% purity) was added. The mixture was stirred for 8 hours under a hydrogen atmosphere and a pressure of 15 psi at 25 °C. The reaction mixture was concentrated under reduced pressure to remove the solvent, giving compound 3 (800 mg, 2.92 mmol, yield 88.7%) as a yellow solid.

[0182] 1H NMR(400MHz,DMSO)δppm 6.74(t,J=9.2Hz,1H)6.24-6.39(m,2H),4.93(s,2H),4.02-4.29(m,2H),3.11(d,J=1 1.6Hz,2H),2.28-2.40(m,1H),1.70(d,J=11.6Hz,2H),1.51(qd,J=12.0,3.2Hz,2H).

[0183] 3. Preparation of 2-((1-(2-fluoro-4-((4-(1-isopropyl-1H-pyrazol-4-yl)-5-methylpyrimidin-2-yl)amino)phenyl)piperidin-4-yl)(methyl-d3)amino)ethane-1,1,2,2-d4-1-ol (compound D010)

[0184] Compound 3 (121 mg, 439 μmol), compound 3A (80.0 mg, 338 μmol), Pd2(dba)3 (19.2 mg, 33.8 μmol), Xantphos (39.1 mg, 67.6 μmol), cesium carbonate (330 mg, 1.01 mmol), and 1,4-dioxane (5 mL) were added sequentially to a reaction flask. The mixture was purged three times with nitrogen, and the reaction was carried out at 100 °C under a nitrogen atmosphere for 2 hours. The reaction solution was concentrated under vacuum to obtain a crude product. The crude product was purified by column chromatography (dichloromethane / anhydrous methanol = 100 / 1 to 7 / 1, dichloromethane / anhydrous methanol = 5 / 1). The crude product was then purified by adding acetonitrile (4 mL) at 70 °C. Compound D010 (55.0 mg, 114 μmol, yield 33.9%, purity 98.9%, deuteration rate 97%) was obtained as a white solid.

[0185] 1 H NMR(400MHz,CHLOROFORM-d)δppm 8.20(s,1H),8.08(d,J=2.8Hz,2H),7.71(dd,J=14.8,2.31Hz,1H),7.10(dd,J=8.4,2.0Hz,1H),7.01(s,1H),6.90-6.96(m,1H),4.58(dt,J=13. 2,6.8Hz,1H),3.46(d,J=11.2Hz,2H),3.04(s,1H),2.62-2.73(m,2H),2 .47-2.58(m,1H),2.37(s,3H),1.79-1.85(m,4H),1.59(d,J=6.8Hz,6H).

[0186] Monitoring LCMS: MS(ESI) m / z = 475.4 [M+H] +

[0187] 4. Preparation of 2-((1-(2-fluoro-4-((4-(1-isopropyl-1H-pyrazol-4-yl)-5-methylpyrimidin-2-yl)amino)phenyl)piperidin-4-yl)(methyl-d3)amino)ethane-1,1,2,2-d4-1-ol formate (compound D010 formate)

[0188] D010 (54 mg, 113.77 μmol) and formic acid (10.93 mg, 227.55 μmol) were dissolved in water (1 mL). The mixture was stirred at 25 °C for 0.5 hours under a nitrogen atmosphere and then freeze-dried. D010 formate (salt ratio 1:1) (59 mg, 113.32 μmol, 99.60% yield) was obtained as a white solid.

[0189] 1 H NMR(400MHz,CHLOROFORM-d)δppm 8.53(s,1H),8.20(s,1H),8.10(s,1H),8.07(s,1H),7.75(dd,J=14.8,2.4Hz,1H),7.10–7.13(m,2H),6.92(t,J=8.8,1H),4.55–4.62(m,1H ),3.50(d,J=12.0,1H),3.10–3.15(m,1H),2.75(t,J=10.8Hz,4H),2.37(s,3H),2.00-2.20(m,2H),1.90-1.95(m,2H),1.60(d,J=6.8,6H).

[0190] Example 9: Preparation of 2-((1-(2-fluoro-4-((5-methyl-4-(1-(prop-2-yl-2-d)-1H-pyrazol-4-yl)pyrimidin-2-yl)amino)phenyl)piperidin-4-yl)(methyl-d3)amino)ethane-1-ol (compound D015)

[0191] 1. Preparation of propan-2-ol-2-d (compound 2)

[0192] Lithium aluminum deuterium (1.70 g, 37.0 mmol) solid was dissolved in tetrahydrofuran (25.0 mL), and acetone (5.00 g, 86.1 mmol) was added dropwise under nitrogen at 0 °C. The mixture was stirred at 0 °C for 1 hour. Lithium aluminum deuterium was quenched by adding Na₂SO₄·10H₂O (5.00 g) to the reaction mixture at 0 °C. The crude product was filtered and used directly in the next production step without concentration or purification. Compound 2 (5.00 g, 81.8 mmol, 95.0% yield) was then obtained in tetrahydrofuran (100 mL).

[0193] 2. Preparation of propionic-2-yl-2-d-4-methylbenzenesulfonate (compound 2A)

[0194] 4-Methylbenzenesulfonyl chloride (7.80 g, 40.9 mmol) and pyridine (24.5 g, 310 mmol) were added separately to a tetrahydrofuran (25 mL) solution and stirred at 25 °C for 16 hours. The reaction solution was quenched with 100 mL of water at 0 °C, and then extracted with 100 mL of ethyl acetate to a final volume of 200 mL. The combined organic layer was washed with 1 M hydrochloric acid (50 x 2 mL), washed with 50 mL of saturated sodium chloride, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and filtered to obtain the residue. The residue was purified by column chromatography (petroleum ether / ethyl acetate = 100 / 1 to 10 / 1, petroleum ether / ethyl acetate = 5 / 1). Compound 2A (2.50 g, 11.6 mmol, 28.4%) was given as a yellow oily liquid.

[0195] 3. Preparation of 2-chloro-5-methyl-4-(1-(propyl-2-yl-2-d)-1H-pyrazol-4-yl)pyrimidine (compound 4)

[0196] A mixture of compound 3 (600 mg, 3.08 mmol), compound 2A (796 mg, 3.70 mmol), and cesium carbonate (3.01 g, 9.25 mmol) in dimethylformamide (10 mL) was purged three times with nitrogen, and then stirred for 2 hours at 90 °C under a nitrogen atmosphere. The reaction mixture was quenched with 20 mL of water, diluted with 20 mL of ethyl acetate, and extracted with 60 mL of dimethylformamide. The combined organic layer was washed with 20 mL of saturated sodium chloride, dried over sodium sulfate, and concentrated by filtration under reduced pressure to obtain the residue. Purification was performed by column chromatography (petroleum ether / ethyl acetate = 100 / 1 to 10 / 1, petroleum ether / ethyl acetate = 5 / 1). Compound 4 (480 mg, 2.02 mmol, yield 65.5%) was given as a white solid.

[0197] 1 H NMR (400MHz, CHLOROFORM-d) δ: 8.34 (s, 1H), 8.20 (s, 1H), 8.05 (s, 1H), 2.45 (s, 3H), 1.57 (s, 6H).

[0198] 4. Preparation of 2-((1-(2-fluoro-4-((5-methyl-4-(1-(prop-2-yl-2-d)-1H-pyrazol-4-yl)pyrimidin-2-yl)amino)phenyl)piperidin-4-yl)(methyl-d3)amino)ethane-1-ol formate (compound D015 formate)

[0199] Compound 5 (150 mg, 555 μmol), compound 4 (120 mg, 505 μmol), Pd₂(dba)₃ (28.6 mg, 50.5 μmol), Xantphos (58.4 mg, 101 μmol), and Cs₂CO₃ (493 mg, 1.51 mmol) were added to dioxane (5 mL), purged three times with nitrogen, and then stirred at 100 °C under a nitrogen atmosphere for 2 hours. The reaction solution was concentrated under vacuum to obtain the crude product. Purification was performed by column chromatography (dichloromethane / anhydrous methanol = 100 / 1–7 / 1, dichloromethane / anhydrous methanol = 5 / 1). The crude product was purified by 4 mL of acetonitrile at 70 °C. Finally, it was purified by preparative separation using high-performance liquid chromatography (mobile phase: formic acid system). The compound D015 formate (salt ratio 1:1) (30.0 mg, 56.97 μmol, yield 11.3%, purity 98.3%, deuteration rate 96%) was obtained as a white solid.

[0200] Preparative separation conditions for high performance liquid chromatography:

[0201] Instrument model: Gilson 281+shamaduzu LC20AP Semi-preparative HPLC system

[0202] Column: CD04-Welch Ultimate C18 150*25mm*7um

[0203] Mobile phase: A is water (0.225% formic acid), B is acetonitrile; Gradient: Initial gradient of mobile phase A 12%, final gradient of mobile phase A 42%, gradient time 10 min; Flow rate: 25 mL / min

[0204] 1 H NMR(400MHz,DEUTERIUM OXIDE)δ:8.42(s,1H),7.55(s,1H),7.48(s,1H),7.40-7.45(m,1H),7.12(dd,J=15.2,2.0Hz,1H),6.81(s,1H),6.69(d,J=8.4Hz,1H),3.94 (t,J=5.2Hz,2H),3.43-3.51(m,1H),3.15-3.43(m,4H),2.68(s,2H),2.16(d,J=10.0Hz,2H),1.84-2.00(m,2H),1.70(s,3H),1.27(s,6H).

[0205] Monitoring LCMS: MS(ESI) m / z = 472.3 [M+H] +

[0206] Example 10: Preparation of 2-((1-(2-fluoro-4-((5-methyl-4-(1-(prop-2-yl-2-d)-1H-pyrazol-4-yl)pyrimidin-2-yl)amino)phenyl)piperidin-4-yl)(methyl)amino)ethane-1-ol (compound D007)

[0207] 1. Preparation of 2-((1-(2-fluoro-4-((5-methyl-4-(1-(prop-2-yl-2-d)-1H-pyrazol-4-yl)pyrimidin-2-yl)amino)phenyl)piperidin-4-yl)(methyl)amino)ethane-1-ol (compound D007)

[0208] Compound 1 (120 mg, 505 μmol), compound 2 (148.46 mg, 555.31 μmol), Pd2(dba)3 (28.6 mg, 50.5 μmol), Xantphos (58.4 mg, 101 μmol), and Cs2CO3 (493 mg, 1.51 mmol) were added to dioxane (5 mL), purged three times with nitrogen, and then stirred at 100 °C under a nitrogen atmosphere for 2 hours. The reaction solution was concentrated under vacuum to obtain a crude product. Purification was performed by column chromatography (dichloromethane / anhydrous methanol = 100 / 1 to 7 / 1, dichloromethane / anhydrous methanol = 5 / 1). The crude product was purified by 4 mL of acetonitrile at 70 °C. Compound D007 (23.2 mg, 49.0 μmol, 9.71% yield, 99.0% purity, 91% deuteration) was obtained as a white solid.

[0209] 1 H NMR (400MHz, CHLOROFORM-d) δ: 8.20 (s, 1H), 8.08 (d, J = 3.2Hz, 2H), 7.71 (dd, J = 14.8, 2.4Hz, 1H), 7.10 (dd, J = 8.8, 2.0Hz, 1H), 6.90-6.97 (m, 2H), 3. 60(t,J=5.2Hz,2H),3.42-3.51(m,2H),2.89-3.17(m,1H),2.63-2.71(m,4 H),2.49-2.60(m,1H),2.32-2.38(m,6H),1.80-1.87(m,4H),1.58(s,6H).

[0210] Monitoring LCMS: MS(ESI) m / z = 469.2 [M+H] +

[0211] 2. Preparation of 2-((1-(2-fluoro-4-((5-methyl-4-(1-(propyl-2-yl-2-d)-1H-pyrazol-4-yl)pyrimidin-2-yl)amino)phenyl)piperidin-4-yl)(methyl)

[0212] (Amino)ethane-1-ol formate (compound D007 formate)

[0213] D007 (24 mg, 51.22 μmol) and formic acid (4.92 mg, 102.44 μmol) were dissolved in water (1 mL). The mixture was stirred at 25 °C for 0.5 hours under a nitrogen atmosphere and then freeze-dried. D007 formate (salt ratio 1:1) (26 mg, 50.52 μmol, 98.64% yield) was obtained as a white solid.

[0214] 1 H NMR(400MHz,CHLOROFORM-d)δppm 8.50(s,1H),8.20(s,1H),8.10(s,1H),8.07(s,1H),7.73(dd,J=14.8,2.4Hz ,1H),7.10(dd,J=8.8,2.0Hz,1H),7.00(s,1H),6.92(t,J=9.2,1H),3.86(t, J=4.8,1H),3.50(d,J=11.6,1H),2.96–3.05(m,3H),2.74(t,J=10.8Hz,2H), 2.69(s,3H),2.37(s,3H),2.00-2.20(m,2H),1.90-1.95(m,2H),1.58(s,6H).

[0215] Example 11: Preparation of 2-((1-(2-fluoro-4-((5-methyl-4-(1-(propyl-2-yl-1,1,1,3,3,3-d6)-1H-pyrazol-4-yl)pyrimidin-2-yl)amino)phenyl)piperidin-4-yl)(methyl-d3)amino)ethane-1-ol (compound D016)

[0216] 1. Preparation of 2-((1-(2-fluoro-4-((5-methyl-4-(1-(propyl-2-yl-1,1,1,3,3,3-d6)-1H-pyrazol-4-yl)pyrimidin-2-yl)amino)phenyl)piperidin-4-yl)(methyl-d3)amino)ethane-1-ol (compound D016)

[0217] Compound 4 (200 mg, 740 μmol), compound 5 (89.8 mg, 370 μmol), Pd₂(dba)₃.CHCl₃ (20.9 mg, 36.9 μmol), Xantphos (42.8 mg, 73.9 μmol), cesium carbonate (361 mg, 1.11 mmol), and 1,4-dioxane (5 mL) were added sequentially to a reaction flask. The mixture was purged three times with nitrogen, and the reaction was carried out at 100 °C under a nitrogen atmosphere for 2 hours. The reaction solution was concentrated under vacuum to obtain a crude product. The crude product was purified by column chromatography (dichloromethane / anhydrous methanol = 100 / 1 to 7 / 1, dichloromethane / anhydrous methanol = 5 / 1). The crude product was then purified by adding acetonitrile at 70 °C. Compound D016 (19.6 mg, 40.4 μmol, yield 10.3%, purity 98.491%, deuteration rate 98%) was obtained as a white solid.

[0218] 1 H NMR(400MHz,CHLOROFORM-d)δppm 8.20(s,1H),8.08(d,J=4.4Hz,2H),7.72(dd,J=14.8,2.4Hz,1H),7.10(dd,J=8.4,1.6Hz,1H),6.86-6.97(m,2H) ,4.56(s,1H),3.61(t,J=4.4Hz,2H),3.47(d,J=11.6Hz,2H),2.52-2.77(m,6H),2.37(s,3H),1.79-1.90(m,4H).

[0219] Monitoring LCMS: MS(ESI) m / z = 477.3 [M+H] +

[0220] 2. Preparation of 2-((1-(2-fluoro-4-((5-methyl-4-(1-(propyl-2-yl-1,1,1,3,3,3-d6)-1H-pyrazol-4-yl)pyrimidin-2-yl)amino)phenyl)piperidin-4-yl)(methyl-d3)amino)ethane-1-ol formate (compound D016 formate)

[0221] D016 (19.4 mg, 40.70 μmol) and formic acid (3.74 mg, 81.40 μmol) were dissolved in water (1 mL). The mixture was stirred at 25 °C for 0.5 hours under a nitrogen atmosphere and then freeze-dried. D016 formate (salt ratio 1:1) (20.0 mg, 38.26 μmol, 94.00% yield) was obtained as a white solid.

[0222] 1H NMR(400MHz,CHLOROFORM-d)δppm 8.53(s,1H),8.20(s,1H),8.10(s,1H),8.07(s,1H),7.75(dd,J=14.8,2.4 Hz,1H),7.12(dd,J=8.8,2.0Hz,1H),7.00(s,1H),6.92(t,J=8.8,1H),4.5 6(s,1H),3.86(t,J=4.8Hz,2H),3.50(d,J=12.0,1H),2.95–3.05(m,4H),2 .75(t,J=10.8Hz,4H),2.37(s,3H),2.00-2.20(m,2H),1.90-1.95(m,2H).

[0223] Example 12: Preparation of 2-((1-(2-fluoro-4-((5-methyl-4-(1-(propan-2-yl-d7)-1H-pyrazol-4-yl)pyrimidin-2-yl)amino)phenyl)piperidin-4-yl)(methyl-d3)amino)ethane-1-ol (compound D017)

[0224] Compound 4 (150 mg, 554 μmol), compound 5 (135 mg, 554 μmol), Pd2(dba)3.CHCl3 (62.8 mg, 111 μmol), Xantphos (128 mg, 222 μmol), cesium carbonate (542 mg, 1.66 mmol), and 1,4-dioxane (5 mL) were sequentially added to a reaction flask. The mixture was purged three times with nitrogen, and the reaction was carried out at 100 °C under a nitrogen atmosphere for 2 hours. The reaction solution was concentrated under vacuum to obtain a crude product. The crude product was purified by column chromatography (dichloromethane / anhydrous methanol = 100 / 1 to 7 / 1, dichloromethane / anhydrous methanol = 5 / 1). The crude product was then purified by adding acetonitrile at 70 °C. Finally, high-performance liquid chromatography (HPLC) was used for preparative separation (mobile phase: formic acid system) to further purify the crude product. The compound D017 formate (salt ratio 1:1) (64.1 mg, 120 μmol, yield 21.7%, purity 98.724%, deuteration rate 98%) was obtained as a white solid.

[0225] Preparative separation conditions for high performance liquid chromatography:

[0226] Instrument model: Gilson 281+shamaduzu LC20AP Semi-preparative HPLC system

[0227] Column: CD04-Welch Ultimate C18 150*25mm*7um

[0228] Mobile phase: A is water (0.225% formic acid), B is acetonitrile; Gradient: Initial gradient of mobile phase A 12%, final gradient of mobile phase A 42%, gradient time 10 min.

[0229] Flow rate: 25 mL / min

[0230] 1 H NMR(400MHz,CHLOROFORM-d)δppm 8.45(s,1H),8.18(s,1H),8.09(d,J=13.2Hz,2H),7.76(dd,J=14.8,2.4Hz,1 H),7.34(s,1H),7.14(dd,J=8.4,1.6Hz,1H),6.92(t,J=9.2Hz,1H),3.95-3. 98(m,2H),3.52(d,J=12.0Hz,2H),3.20-3.32(m,1H),3.04-3.13(m,2H),2.7 7(t,J=11.2Hz,2H),2.38(s,3H),2.16(d,J=11.6Hz,2H),1.90-2.04(m,2H).

[0231] Monitoring LCMS: MS(ESI) m / z = 478.5 [M+H] +

[0232] Example 14: Preparation of 2-((1-(2-fluoro-4-((4-(1-isopropyl-1H-pyrazol-4-yl)-5-methylpyrimidin-2-yl)amino)phenyl)piperidin-4-yl)(methyl)amino)ethane-1-ol (control compound D000)

[0233] Compound 1d (0.8 kg, 3 mol), compound 1e (0.92 kg, 3.9 mol), Pd2(dba)3 (137 g, 0.15 mol), Xantphos (174 g, 0.3 mol), cesium carbonate (2.5 kg, 7.5 mol), and 1,4-dioxane (20 L) were added sequentially to a reaction flask. Under nitrogen protection, the mixture was heated to an internal temperature >100 °C and reacted for 10 hours. After the reaction was complete, the mixture was filtered through diatomaceous earth. The filtrate was concentrated under reduced pressure. 6 L of dichloromethane and approximately 6 L of 2.4% hydrochloric acid aqueous solution were added sequentially to the concentrated residue to adjust the pH to 4–5. The mixture was then separated. The aqueous phase was washed three times with dichloromethane (6L x 3 times). A 13% potassium hydroxide aqueous solution (about 3L) was added to the aqueous phase to adjust the pH to 13-14. Then, 10L of dichloromethane was added and the mixture was separated. The organic phase was concentrated under reduced pressure to obtain compound D000 (60% yield, 99.9% purity).

[0234] 1H NMR(400MHz, DMSO-d6)δ:9.38(s,1H),8.31(d,J=20.6Hz,2H),8.08(s,1H),7.75(dd,J=15.5,2.4Hz, 1H),7.47–7.40(m,1H),6.99(dd,J=10.1,8.8Hz,1H),4.63(p,J=6.7Hz,1H),4.33(s,1H),3.45(t,J=6 .5Hz,2H),3.30(d,J=11.7Hz,2H),2.65–2.55(m,2H),2.43(td,J=11.6,5.9Hz,1H),2.31(s,3H),2.2 4(s,3H),1.81–1.72(m,2H),1.58(qd,J=12.2,3.9Hz,2H),1.48(d,J=6.6Hz,6H).m / z:468.2881[M+H] + .

[0235] Example 1: Pharmacokinetic Evaluation in Rats

[0236] Weigh compounds D000 (control compound), D001, D004, D008, D009, D010, and D027, add a small amount of DMSO, and then add sodium chloride solution for injection to prepare a solution of 1.5 mg / mL. -1 Solutions of compounds D000, D001, D008, D009, D010, and D027 are prepared for administration.

[0237] 21 male rats, at 7.5 mg / kg -1 After oral administration, approximately 0.03 mL of blood was collected at 5 min, 15 min, 30 min, 1 h, 2 h, 4 h, 6 h, 8 h, and 24 h post-administration. The blood was centrifuged at 3500 rpm for 15 min, and the supernatant plasma was collected. 5 μL of plasma was placed in an EP tube, and 100 μL of acetonitrile containing 20 ng·ml⁻¹ internal standard SAHA was added to precipitate the protein. The tube was vortexed for 30 s and centrifuged at 13000 rpm for 15 min. The supernatant was then collected and placed in a sample vial for analysis.

[0238] Standard curve range: 1–1000 ng / ml -1 .

[0239] The concentration of the compounds of the present invention in plasma was determined by LC-MS / MS. Pharmacokinetic parameters were calculated based on the plasma drug concentrations of each animal at different time points, as shown in Table 1. Analysis of the results shows that, compared to the control compound D000 of Example 12, the compounds of the present invention (compounds of Examples 1-11) exhibit better pharmacokinetics in animals, and therefore better pharmacodynamics and therapeutic effects.

[0240] Table 1. Pharmacokinetic parameters of rats after gavage administration of 7.5 mg / kg

[0241] The Cmax (230±42ug / L) and AUC(0-t) (2551±98ug / L*h) of compound D004, the Cmax (325±39ug / L) and AUC(0-t) (3224±317ug / L*h) of compound D008, the Cmax (298±42ug / L) and AUC(0-t) (3001±456ug / L*h) of compound D009, and the Cmax (277±20ug / L) and AUC(0-t) (2969±254ug / L*h) of compound D027 were all significantly higher than those of the control compound D000 (225±327ug / L) and AUC(0-t) (1999±325ug / L*h).

[0242] Example 2: In vitro pharmacodynamic evaluation of the compounds of the present invention against JAK kinase and FLT kinase

[0243] Inhibition of kinases JAK2 and FLT3 IC 50 Measurement

[0244] Kinase activity inhibition assays were performed in 384-well plates. Test compounds (D000, D008, and D009) were dissolved in DMSO to prepare test solutions of different concentrations. Kinases were added to kinase buffer (50 mM HEPES, pH 7.5, 10 mM MgCl2, 0.01% Tween-20, 0.01% BSA, 2 mM DTT) to form an enzyme solution. The test compound solution and ATP were then added to the enzyme solution, and the mixture was incubated at room temperature for 60 minutes. Finally, a stop reaction solution containing antibody was added, and the mixture was incubated at room temperature for another 60 minutes. The Envision 2104 Multilabel Reader reads the emission values ​​at 340 nm excitation and 520 nm emission at 495 nm. The ratio of the fluorescence readings (Lantha signal (520 nm / 495 nm)) is replicated and converted into the inhibition percentage using the formula: Percent inhibition = (max - Lantha signal) / (max - min) * 100. "min" represents the reading of the control wells without enzyme; "max" represents the reading of the control wells with DMSO. The final IC50 value is then calculated. 50 The values ​​are shown in Table 2, and the structures are displayed. The compounds described in this invention exhibit excellent inhibitory activity against both kinases JAK2 and FLT3.

[0245] Table 2

Claims

1. A compound of formula (I), a solvate thereof, a pharmaceutically acceptable salt thereof, or a solvate of a pharmaceutically acceptable salt thereof, in, L is -(CR) a R b ) n -, R a and R b Independently hydrogen or deuterium, n is 2, 3, 4, 5 or 6; R 1 is hydrogen, deuterium or Y 1 , Y 6 , Y 7 , Y 8 , Y 9 , Y 10 , Y 11 , Y 12 , Y 13 , Y 14 , Y 15 , Y 16 , Y 17 , Y 18 , Y 19 , Y 20 , Y 21 , Y 22 , Y 23 , Y 24 , Y 25 , Y 26 , Y 27 , Y 28 , Y 29 , Y 30 , Y 31 , Y 32 , Y 33 and Y 34 are each independently hydrogen or deuterium; and said L, R 1 , Y 1 , Y 6 , Y 7 , Y 8 , Y 9 , Y 10 , Y 11 , Y 12 , Y 13 , Y 14 , Y 15 , Y 16 , Y 17 , Y 18 , Y 19 , Y 20 , Y 21 , Y 22 , Y 23 , Y 24 , Y 25 , Y 26 , Y 27 , Y 28 , Y 29 , Y 30 , Y 31 , Y 32 , Y 33 and Y 34 at least one of which comprises deuterium.

2. The compound as shown in formula (I), solvate thereof, pharmaceutically acceptable salt thereof or solvate of the pharmaceutically acceptable salt thereof according to claim 1, wherein, It meets one or more of the following conditions: ​ (1) Y 1 , Y 9 , Y 10 , Y 15 , Y 16 , Y 17 , Y 18 , Y 19 , Y 20 , Y 21 , Y 22 , Y 23 , Y 24 , Y 25 , Y 26 and Y 34 are hydrogen; (2) L is n1 is 0, 1, 2, 3, or 4, with the "1" position connected to an oxygen atom; Y 2 Y 3 Y 4 and Y 5 Independently hydrogen or deuterium; preferably, L is For example, L is (3)R 1 for For example, methyl or (4) for (5) for (6) for (7) The pharmaceutically acceptable salt is a formate, for example, 1 molecule of formate; (8) The content of deuterium isotopes at the deuterium substitution sites is 30% or more, preferably 50% or more, more preferably 75% or more, more preferably 95% or more, more preferably 96% or more, more preferably 97% or more, more preferably 98% or more, and more preferably 99% or more.

3. The compound of formula (I) as claimed in claim 1 or 2, its solvates, pharmaceutically acceptable salts thereof, or solvates of pharmaceutically acceptable salts thereof, characterized in that, The compound represented by formula (I) is a compound represented by formula I-1, I-2, I-3, I-4, I-5, I-6, I-7, I-8, I-9, I-10, I-11, I-12 or I-13:

4. The compound of formula (I) as claimed in claim 3, its solvates, pharmaceutically acceptable salts thereof, or solvates of pharmaceutically acceptable salts thereof, characterized in that: The compound shown in formula (I) is the same as that shown in formula I-4-1. Alternatively, the compound shown in formula (I-12) is the compound shown in formula I-12-1.

5. The compound of formula (I) as claimed in claim 1, its solvates, pharmaceutically acceptable salts thereof, or solvates of pharmaceutically acceptable salts thereof, characterized in that, The compound shown in formula (I) is any of the following compounds: The pharmaceutically acceptable salt of the compound shown in formula (I) is any of the following compounds: Preferably, the formate is a single molecule of formate.

6. A pharmaceutical composition comprising: (1) The compound of formula (I) as claimed in any one of claims 1-5, its solvates, pharmaceutically acceptable salts thereof, or solvates of pharmaceutically acceptable salts thereof; and, (2) Pharmaceutical excipients.

7. The use of a compound of formula (I) as described in any one of claims 1-5, a solvate thereof, a pharmaceutically acceptable salt thereof, a solvate thereof, or a pharmaceutical composition as described in claim 6 in the preparation of JAK2 kinase and / or FLT3 kinase inhibitors.

8. The use of a compound of formula (I) as claimed in any one of claims 1-5, a solvate thereof, a pharmaceutically acceptable salt thereof, a solvate thereof, or a pharmaceutical composition as claimed in claim 6 in the preparation of a medicament for treating and / or preventing diseases mediated by JAK2 kinase and / or FLT3 kinase.

9. The application as described in claim 8, characterized in that: The diseases mediated by JAK2 kinase and / or FLT3 kinase are hematologic malignancies, such as acute myeloid leukemia, chronic myeloid leukemia, myeloma, acute lymphoblastic leukemia, acute myeloid leukemia, acute promyelocytic leukemia, chronic lymphocytic leukemia, chronic neutrophilic leukemia, acute undifferentiated cell leukemia, myelodysplastic syndrome, myelodysplastic syndrome, myelofibrosis, multiple myeloma, spinal sarcoma, or primary thrombocytosis.

10. The use of a compound of formula (I) as described in any one of claims 1-5, a solvate thereof, a pharmaceutically acceptable salt thereof, a solvate thereof, or a pharmaceutical composition as described in claim 6 in the preparation of a medicament for treating and / or preventing hematologic malignancies; Preferably, the hematologic malignancy is acute myeloid leukemia, chronic myeloid leukemia, myeloma, acute lymphoblastic leukemia, acute myeloid leukemia, acute promyelocytic leukemia, chronic lymphocytic leukemia, chronic neutrophilic leukemia, acute undifferentiated cell leukemia, myelodysplastic syndrome, myelodysplastic syndrome, myelofibrosis, multiple myeloma, spinal cord sarcoma, or primary thrombocytosis.

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