CN114573591B

CN114573591B - Substituted pyrrolopyrimidine compound and application thereof

Info

Publication number: CN114573591B
Application number: CN202210398671.2A
Authority: CN
Inventors: 曾燕群; 黄龙; 周广林; 朱绪成; 付海霞; 牟霞
Original assignee: Chengdu Shibeikang Biological Medicine Technology Co ltd
Current assignee: Chengdu Shibeikang Biological Medicine Technology Co ltd
Priority date: 2022-04-16
Filing date: 2022-04-16
Publication date: 2023-04-25
Anticipated expiration: 2042-04-16
Also published as: CN114573591A

Abstract

The invention discloses a pyrrolopyrimidine compound shown in a formula (I), a stereoisomer or a pharmaceutically acceptable salt thereof. The invention also provides a preparation method of the compound, a stereoisomer or a pharmaceutically acceptable salt thereof, and application of the compound in preparing a medicament for preventing or treating C-Kit and/or PDGFR, in particular mutant C-Kit and/or mutant PDGFR mediated diseases. The compound has better pharmacokinetic property and stronger clinical medication compliance.

Description

Substituted pyrrolopyrimidine compound and application thereof

Technical Field

The invention belongs to the field of medicines, and particularly relates to a substituted pyrrolopyrimidine compound, a preparation method thereof and application thereof in preventing and treating C-Kit and/or PDGFR mediated diseases.

Background

KIT belongs to a member of the family of type III receptor tyrosine kinases encoded by proto-oncogenes C-KIT. The KIT protein coded by the C-KIT gene consists of an intracellular tyrosine kinase region, a transmembrane region and an extracellular region with a ligand binding site, and after the KIT receptor is combined with a ligand SCF (stem cell factor), downstream signals including Ras, raf, MAPK channels and the like are activated through the formation of dimers, and finally transcription factors in cells are activated, so that gene expression is regulated, and cell growth and proliferation are controlled.

Imatinib is a tyrosinase inhibitor, and can block the function of tyrosine kinase KIT receptor, thereby inhibiting tumor formation. Studies have demonstrated that the location of KIT mutations can affect the response of tumor patients to imatinib. The KIT mutation is located in a regulatory region, namely a non-tyrosinase structural region, so that the inhibitor can effectively block enzyme sites, the partial remission rate of imatinib is obviously improved, the average life cycle is prolonged, and the disease progress is slow.

Such as: gastrointestinal stromal tumor is a relatively common malignant gastrointestinal tumor. The pathological research proves that the kinase c-Kit is an effective target point for treating the gastrointestinal stromal tumor, plays an important role in the transfer and differentiation process of cells, and the over-expression of the kinase c-Kit is closely related to the gastrointestinal stromal tumor. Currently, imatinib is the first line of clinical treatment for gastrointestinal stromal tumors, but after long-term administration, approximately 80-85% of patients develop resistance, the main factor of resistance being the development of a compound for inhibiting wild-type C-Kit and mutant C-Kit is highly desirable for clinical treatment because of the drug-resistant mutation of C-Kit kinase.

Patent CN201810040590 discloses a C-Kit inhibitor and application thereof, and the disclosed embodiment of the compound III has strong inhibition effect on wild type C-Kit (WT), mutant C-Kit (D861V) and PDGFR alpha (D842V) kinase; not only has therapeutic effect on the diseases mediated by the wild type of the c-Kit, but also has good effect on the diseases mediated by the mutated c-Kit. However, this compound is metabolized faster in vivo and longer acting C-Kit inhibitors are clinically needed.

Disclosure of Invention

In view of the shortcomings of the prior art, it is an object of the present invention to provide a deuterated pyrrolopyrimidine compound having a relatively high half-life useful for the prevention and treatment of C-Kit and/or PDGFR mediated diseases including gastrointestinal stromal tumors, lupus, leukemia, mastocytosis, melanoma, seminoma, rheumatoid arthritis, multiple sclerosis, freezing syndrome, multiple myeloma, pancreatic cancer and the like.

The invention provides a compound shown as a formula (I), a stereoisomer or a pharmaceutically acceptable salt thereof:

wherein R1, R2, R3, R4, R5, R6 are independently hydrogen or deuterium, and R1, R2, R3, R4, R5, R6 are not all hydrogen.

Further, the compound shown in the formula (I), a stereoisomer or a pharmaceutically acceptable salt thereof has the following structure:

wherein, at least three of R1, R2, R3, R4, R5 and R6 are deuterium, and the rest are hydrogen.

Further, the compound represented by the above formula (I), a stereoisomer or a pharmaceutically acceptable salt thereof, has the structure as follows:

、

or (b)

。

Further, the compound of formula (I), a stereoisomer or a pharmaceutically acceptable salt thereof, has the structure:

or->

。

or->

。

Further, the above compound, stereoisomer or pharmaceutically acceptable salt thereof, characterized in that the salt includes, but is not limited to, acetate, ascorbate, benzoate, benzenesulfonate, citrate, fumarate, hydrochloride, hydrobromide, maleate, methanesulfonate, nitrate, oxalate, phosphate, succinate or sulfate.

Further, the above-mentioned compounds also include nitrogen oxides, hydrates, solvates or metabolites thereof.

Further, the invention provides a pharmaceutical composition containing the compound, the stereoisomer, the nitrogen oxide, the hydrate, the solvent compound, the metabolite or the pharmaceutically acceptable salt thereof, and the composition further comprises a pharmaceutically acceptable carrier and/or auxiliary materials.

Further, the pharmaceutically acceptable carriers or excipients include, but are not limited to, oral formulation excipients or parenteral administration excipients, and the administration route may be oral, injection, etc.; the administration dosage form can be liquid dosage form, solid dosage form, liquid dosage form can be syrup, injection solution, nonaqueous solution, suspension or emulsion, and solid dosage form can be tablet, lozenge, capsule, dripping pill, granule, powder, cream, solution, suppository, dispersible powder such as lyophilized powder for injection, aerosol, etc.; the excipients used include, but are not limited to: lactose, calcium carbonate, calcium phosphate, sodium phosphate, starch, cyclodextrin, sucrose, mannitol, microcrystalline cellulose sodium, calcium sulfate, water, ethanol, propanol, glycerol, propylene glycol, isopropanol, glucose, sodium carboxymethyl cellulose, potassium phosphate, dried starch, agar powder, calcium carbonate, sodium bicarbonate, sodium dodecyl sulfate, methylcellulose, glyceryl tristearate, hydrogenated oil, talc, magnesium triethylamine stearate, silica, corn starch, stearate, boric acid, liquid paraffin.

Further, the invention provides the use of the above compound, stereoisomer, nitrogen oxide, hydrate, solvent compound, metabolite or pharmaceutically acceptable salt thereof in the preparation of a medicament for preventing or treating a C-Kit and/or PDGFR mediated disorder.

Still further, the above-described C-Kit and/or PDGFR mediated diseases include gastrointestinal stromal tumors, lupus, leukemia, mast cell disease, melanoma, seminoma, rheumatoid arthritis, multiple sclerosis, freezing syndrome, multiple myeloma or pancreatic cancer.

Further, the C-Kit and/or PDGFR is a mutant C-Kit and/or PDGFR.

The beneficial effects are that: the compound has better pharmacokinetic property, slower metabolism, longer half-life period and longer time for playing the drug effect, is more suitable for clinical medication, and provides better choice for tumor patients.

Detailed Description

The present invention will be described in further detail with reference to the following examples, which are only for illustrating the technical aspects of the present invention and are not intended to limit the present invention, and any equivalent substitutions in the art according to the present disclosure are within the scope of the present invention.

The compounds of the present invention, stereoisomers or pharmaceutically acceptable salts thereof may be prepared by the synthetic routes of the examples, and the conventional conditions of the reaction starting materials and reaction solvents may be adjusted according to the substituents or salt-forming requirements, which may be accomplished by one skilled in the art based on the present disclosure. In addition, the column chromatography of the present invention refers to silica gel column chromatography unless otherwise specified, and the eluting solvent may be a single or mixed eluting solvent determined by combining the reaction solvent with common knowledge or common means of a person skilled in the art.

The structure of the compound is changed into a nuclear magnetic resonance structure ¹ H NMR) or liquid mass spectrometry (LC-MS).

The liquid chromatography-mass spectrometer (LC-MS) is Agilent G6120B (matched with liquid phase Agilent 1260); nuclear magnetic resonance apparatus ¹ H NMR) of Bruker AVANCE-300, nuclear magnetic resonance ¹ H NMR) shift [ ]δ) Given in parts per million (ppm), the assay solvent is DMSO, the internal standard is Tetramethylsilane (TMS), and the chemical shift is 10 ^~6 (ppm) is given as a unit.

The term "room temperature" according to the invention means a temperature between 10 and 25 ℃.

Example 1: preparation of Compound 1

Step 1: synthesis of Compound c

To a single bottle was added compound a (2.86 g,10 mmol) and anhydrous THF (200 mL), the solution was stirred, compound b (2.42 g,20 mmol) and tetraethyltitanate (6.83 g,30 mmol) were added, and the temperature was raised to 70 ℃ under nitrogen protection, and stirred overnight. After completion of the TLC monitoring reaction, water (100 mL) was added, ethyl acetate extraction (300 mL) was performed three times, the organic phases were combined, washed twice with water, washed twice with saturated sodium chloride, dried over anhydrous sodium sulfate, filtered, and concentrated. The product was separated by column chromatography to give a yellow solid 2.43, g. Yield: 62.5 Percent of the total weight of the composition.

MS：m/z (ES): 390.5[M+1]。

Step 2: synthesis of Compound D

Magnesium powder (280 mg,11.6 mmol) was added to a 50mL two-necked flask, the flask was evacuated and purged with nitrogen, diethyl ether (10 mL) was added, deuterated iodomethane CD3I (1.44 g,10 mmol) was added dropwise, and after completion of the dropwise addition, the temperature was raised to reflux and the reaction was maintained under stirring for 2 hours. Cooled to room temperature.

Compound c (778 mg, 2.0 mmol) and anhydrous THF (30 mL) were added to another 50mL two-necked flask, stirred to clarify, evacuated and protected with nitrogen, cooled to 0 ℃, slowly added to the solution, and after dropwise addition of the prepared solution of CD3MgI in diethyl ether, stirring was continued to react for 2 hours at 0 ℃. The reaction was quenched by addition of saturated aqueous ammonium chloride (25 mL), extracted with ethyl acetate (50 mL x 3), the combined organic phases were washed with water (50 mL), saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered, concentrated and passed through a column to give a white solid 511 mg in 62.6% yield.

MS：m/z (ES):409.5[M+1]。

Step 3: synthesis of Compound E

Compound D (408 mg,1.0 mmol) and methanol (15 mL) were added to a 50mL magnetically-stirred two-necked flask, stirred until clear, and then a hydrogen chloride dioxane solution (12 mL, 8M) was added, followed by stirring at room temperature under nitrogen atmosphere for 1 hour. The solvent was evaporated under reduced pressure, dichloromethane (30 mL) and saturated aqueous sodium bicarbonate (20 mL) were added, stirred for 2 min, the organic layer was separated, the aqueous phase was extracted with dichloromethane (30 ml x 2), the organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated to give 212mg of a white solid in 69.7% yield.

MS：m/z (ES):305.5[M+1]。

Step 4: synthesis of Compound 1

Compound E (200 mg, 0.66 mmol) and 1, 4-dioxane (10 mL) were added to a 50mL single neck flask equipped with a magnetic stirrer, stirred until clear, DIPEA (131 mg,1 mmol) and compound f (128 mg,0.55 mmol) were added and the reaction was stirred at room temperature under nitrogen overnight. The solvent was evaporated under reduced pressure, and the residue was purified by column chromatography on silica gel to give 255, 255 mg as a white solid in 92.8% yield.

MS：m/z (ES): 502.5[M+1]。

¹ H NMR(300MHz, DMSO-d6): 8.58 (m,2H), 8.22 (m,1H),7.91-7.96 (m,2H), 7.19-7.28 (m,4H), 6.41(s,1H), 5.10 (t,2H), 5.01(s,1H)，3.95(m,3H)，3.26 (m,4H)，3.18-3.22（(m,4H)。

Example 2: preparation of Compound 2

100mg of Compound 1 was taken and isolated by chiral HPLC to give 40mg of a white solid as isomer Compound 2.

MS m/z (ES): 502.5[M+1]。

¹ H NMR(300MHz, DMSO-d6): 8.59 (m,2H), 8.21 (m,1H),7.91-7.96 (m,2H), 7.19-7.28 (m,4H), 6.41(s,1H), 5.10 (t,2H), 5.01(s,1H)，3.95(m,3H)，3.26 (m,4H)，3.18-3.21（(m,4H)）。

Example 3: preparation of Compound 3

100mg of Compound 1 was taken and isolated by chiral HPLC to give 40mg of a white solid as isomer 3.

MS m/z (ES): 502.5[M+1]。

¹ H NMR(300MHz, DMSO-d6): 8.59 (m,2H), 8.21 (m,1H),7.91-7.96 (m,2H), 7.19-7.28 (m,4H), 6.41(s,1H), 5.10 (t,2H), 5.01(s,1H)，3.95(m,3H)，3.26 (m,4H)，3.18-3.21(m,4H)。

Example 4: preparation of Compound 4

Step 1: synthesis of Compound G

Compound h (10.0 g,51.5 mmol) and anhydrous THF (50 mL) were dissolved with stirring in a 150mL two-necked flask equipped with a magnetic stirrer, naH (4.5g,103.0 mmol,55% w/w) was added, followed by stirring under nitrogen for 10 minutes, then CD3I (14.9 g,103.0 mmol) was added dropwise, the ice bath was removed after the dropwise addition, and the reaction was stirred at room temperature under nitrogen overnight. The reaction was quenched by the addition of methanol (10 mL), diluted with ethyl acetate (50 mL), the insoluble material was filtered off, the filtrate was concentrated, and passed through a silica gel column to give 6.6g of a colorless oil in 60.7% yield.

MS：m/z (ES): 212.1[M+1]。

Step 2: synthesis of Compound K

Compound j (2.14G, 10 mmol), compound G (4.22G, 20 mmol), cesium carbonate (9.77G, 30 mmol), pd (dppf) Cl ₂ . CH ₂ Cl ₂ (816 mg,1 mmol) was placed in a 100mL single neck flask equipped with a magnetic stirrer, 1, 4-dioxane (80 mL), ethanol (15 mL) and water (10 mL), evacuated and replaced 3 times with nitrogen, heated to 110℃overnight under nitrogen protection, cooled to room temperature, concentrated to dryness under reduced pressure, and column chromatographed to give a yellow solid 1.36 g in 62.3% yield.

MS：m/z (ES):219.2[M+1]。

Step 3: synthesis of Compound F

Compound K (1.36 g,6.23 mmol) and phosphorus oxychloride (13 mL) were added to a 50mL single-necked flask, heated to 95 ℃ under nitrogen protection, and reacted with stirring for 5 hours. After cooling to room temperature, the remaining phosphorus oxychloride was distilled off under reduced pressure, methylene chloride (30 mL) and saturated aqueous sodium hydrogencarbonate (10 mL) were added, the organic layer was separated, the aqueous layer was extracted with methylene chloride (20 mL) to give an organic phase, which was dried over anhydrous sodium sulfate, filtered, concentrated, and subjected to column chromatography to give 476. 476 mg as a white solid in 31.3% yield.

MS：m/z (ES): 237.5[M+1]。

Step 4: synthesis of Compound d

Magnesium powder (280 mg,11.5 mmol) was added to a 50mL two-necked flask, evacuated and purged with nitrogen, diethyl ether (10 mL) was added under high pressure, and methyl iodide (1.42 g,10 mmol) was added dropwise. After the dripping is finished, the temperature is increased to reflux, and the stirring reaction is kept for 2 hours. Cooled to room temperature.

Compound c (778 mg, 2.0 mmol) and anhydrous THF (30 mL) were added to another 50mL two-necked flask, stirred to clarify, evacuated and protected with nitrogen, cooled to 0 ℃, slowly added dropwise to the prepared CH3MgI diethyl ether solution, stirred continuously, and reacted at 0 ℃ for 2 hours. The reaction was quenched by addition of saturated aqueous ammonium chloride (25 mL), extracted with ethyl acetate (50 mL x 3), the combined organic phases were washed with water (50 mL), saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered, concentrated and chromatographed on a column to give 511 mg as a white solid in 63.1% yield.

MS：m/z (ES):406.5[M+1]。

Step 5: synthesis of Compound e

Compound d (408 mg,1.0 mmol) and methanol (15 mL) were added to a 50mL magnetically-stirred two-necked flask, stirred until clear, and then a hydrogen chloride dioxane solution (12 mL, 8M) was added, followed by stirring at room temperature under nitrogen atmosphere for 1 hour. The solvent was distilled off under reduced pressure, methylene chloride (30 mL) and saturated aqueous sodium hydrogencarbonate solution (20 mL) were added, stirred for 2 minutes, the organic layer was separated, the aqueous phase was extracted with methylene chloride (30 ml×2), and the organic phase was combined, dried over anhydrous sodium sulfate, filtered and concentrated to give 212mg of a white solid in 70.3% yield.

MS：m/z (ES):302.3[M+1]。

Step 6: synthesis of Compound 4

Compound e (200 mg, 0.66 mmol) and 1, 4-dioxane (10 mL) were added to a 50mL single neck flask equipped with a magnetic stirrer, stirred until clear, DIPEA (131 mg,1 mmol) and compound F (130 mg,0.55 mmol) were added and the reaction was stirred at room temperature overnight under nitrogen. The solvent was evaporated under reduced pressure, and the residue was purified by column chromatography on silica gel to give 252. 252 mg as a white solid in 91.5% yield.

MS：m/z (ES): 502.2[M+1]。

¹ H NMR：(300MHz，DMSO-d6):8.59(m，2H)，8.21(m，1H)，7.91-7.96(m，2H)，7.19-7.28(m，4H)，6.41(s，1H)，5.10(t，2H)，5.01(s，1H)，3.26(m，4H)，3.18-3.21(m，4H)，1.76(s，3H)。

Example 5: preparation of Compound 5

100mg of Compound 4 were taken and isolated by chiral HPLC to give 40mg of a white solid as isomer compound 5.

MS m/z (ES): 502.2[M+1]。

Example 6: preparation of Compound 6

100mg of Compound 4 were taken and isolated by chiral HPLC to give 40mg of a white solid as isomer 6.

MS m/z (ES): 502.2[M+1]。

Example 7: preparation of Compound 7

Compound E (200 mg, 0.66 mmol) and 1, 4-dioxane (10 mL) were added to a 50mL single neck flask equipped with a magnetic stirrer, stirred until clear, DIPEA (131 mg,1 mmol) and compound F (128 mg,0.54 mmol) were added and the reaction was stirred at room temperature under nitrogen overnight. The solvent was evaporated under reduced pressure, and the residue was purified by column chromatography on silica gel to give 255, 255 mg as a white solid in 93.4% yield.

MS：m/z (ES): 505.5[M+1]。

¹ H NMR：(300MHz，DMSO-d6):8.59(m，2H)，8.21(m，1H)，7.91-7.96(m，2H)，7.19-7.28(m，4H)，6.41(s，1H)，5.10(t，2H)，5.01(s，1H)，3.26(m，4H)，3.18-3.21(m，4H)。

Example 8: preparation of Compound 8

100mg of Compound 7 was taken and isolated by chiral HPLC to give 42mg of a white solid as isomer 8.

MS m/z (ES): 505.5[M+1]。

Example 9: preparation of Compound 9

100mg of Compound 7 was taken and isolated by chiral HPLC to give 42mg of a white solid as isomer 9.

MS m/z (ES): 505.5[M+1]。

Comparative example: synthesized according to the preparation method disclosed in patent CN201810040590

。

Test example 1: in vitro enzyme inhibition Activity assay

Test principle:

detection of chemiluminescence in Assay plates by Envision as compound ICs ₅₀ The inhibition of c-Kit (WT), c-Kit mutant D861V and PDGFR alpha mutant D842V kinases by the compounds was evaluated using the values as indicators.

The test method comprises the following steps:

the comparative and example compounds 2, 3, 5, 8 were diluted in DMSO to 1mM for use. Before use, each test compound was diluted 25-fold with ultrapure water and equilibrated at room temperature for 30min for use. In an Assay plate, each test compound was subjected to 3-fold gradient dilution to obtain 11 concentration gradients with final concentrations ranging from 1uM to 0.017 nM; adding a c-Kit (WT) or c-Kit (D861V) or PDGFR alpha (D842V) kinase mixture into an Assay plate, incubating for 30min at room temperature, adding a substrate ATP mixture to start a reaction, incubating for 90min at room temperature, adding a stop solution to stop the reaction, incubating for 1 h at room temperature, reading a plate, analyzing data, and calculating the IC of the compound ₅₀ Values. Wherein A represents IC ₅₀ <10 nM, B represents 10 nM< IC ₅₀ < 100 nM。

As is clear from Table 1, the comparative example compounds and example compounds 2, 3, 5 and 8 have strong inhibitory effects on wild-type c-Kit (WT), mutant c-Kit (D861V) and PDGFRa (D842V) kinases.

Test example 2: in vitro liver microsome stability test

Test materials

Medicine: comparative and example compounds 1, 2, 3, 5, 8;

reagent: methanol, acetonitrile, formic acid as chromatographic purity, purchased from MERCK company; other reagents were all commercially available analytical pure; the experimental water is ultrapure water;

materials: human liver microsomes were purchased from Celsis/rad liver disease research limited (Shanghai); NADPH was purchased from Toronto Research Chemicals Inc (Ontario, canada).

Test procedure

Test of metabolic stability of comparative and example Compounds 1, 2, 3, 5, 8 in human liver microparticles

(1) Preparation of stock solution

A) Liver particles: 20 mg/mL;

b) Sodium phosphate buffer: 200 mM, pH 7.4;

c) Testosterone (Yang Can): 5 mM;

D)NADPH：10 mM；

e) Stop solution: acetonitrile (containing internal standard 40 ng/mL bromobuterol);

f) Test article: 2 mM.

(2) Reaction system (final volume, concentration)

Total incubation volume: 1 mL;

sodium phosphate buffer: 0.1 M, pH 7.4;

liver microparticle protein concentration: 0.20 mg/mL;

test article concentration: 1. Mu.M;

testosterone (Yang Can) concentration: 10. Mu.M;

NADPH concentration: 1 mM;

reaction time: 0,5, 15, 30, 50 min.

(3) Reaction and termination

The reaction system was pre-incubated at 37℃for 5 min, and then NADPH was added to initiate the reaction. The negative control group was added to the system with sodium phosphate buffer instead of NADPH. 100. Mu.L of the reaction solution was extracted according to the above time and mixed with 300. Mu.L of a termination solution containing the internal standard bromobute Luo Lengdong to terminate the reaction. The supernatants were then centrifuged at 4℃and the amount of compound was measured by LC-MS/MS at each time point. 2 replicates were taken at each time point. A positive control group with testosterone as a substrate was also set.

Instrument for measuring and controlling the intensity of light

Liquid phase-mass spectrometry analysis system (LC-MS/MS) comprising a senior company VP series Nanospace S1-2 3301 binary pump and Nanospace S1-2 3202 online degasser (japan), nanospace 51-2 3133 multipurpose autosampler and AB company API-4000Q-Trap mass spectrum detector (containing ESI ion source), column: venusil MP-C18 (2.1X10 mm, 3 μm).

Chromatographic conditions

Mobile phase a was an aqueous phase, 5% methanol (containing 0.1% formic acid); mobile phase B was the organic phase, 95% methanol (containing 0.1% formic acid). The flow rate is 0.3 mL min ^-1 Column temperature: the gradient was used at room temperature as shown in the following table.

。

Mass spectrometry conditions

。

Test results

As shown in Table 4, compounds 1, 2, 3, 5, 8 had much higher percent parent residues than the comparative compounds at 60min in human liver microsomes. Illustrating that the example compounds of the present invention metabolize slower, potentially longer acting in vivo than the comparative compounds.

Test example 3: rat pharmacokinetic test

Test materials

Compounds 2, 5, 8 and comparative compounds.

Main instrument and equipment

Liquid mass spectrometer, manufacturer: waters; model: ACQUITY UPLC I-class+XeFo TQ-S.

Test system

(1) Variety/strain/grade

Strain: SD rats; grade: SPF stage.

(2) Sex and number

Number and sex of animals entering acclimation period: 40, male and female halves;

number and sex of animals used: 32, male and female halves.

(3) Weight and age

Weight of: 136.9-148.5 g of female, 137.2-158.5 g of male, 186.0-207.8 g of female, 267.3-303.9 g of male when grouped, wherein the weight of the individuals is within the average weight of the same sex of +/-20%;

age: the purchasing time is 5-6 weeks, and the test grouping time is 7-9 weeks.

The test was divided into 4 groups (single groups of example compounds 2, 5, 8 and comparative example compounds), 8 animals/group, each male and female, and in order to prevent the occurrence of the situation that the purchased experimental animals cannot meet the test requirement due to unknown reasons, 2 animals/sex were additionally purchased for a total of 40 SD rats (32 actual use).

Animal grouping information is shown in the following table:

note that: the first digit of the animal number represents the group (1, 2, 3, 4 represent test 1, 2, 3, and comparative group, respectively); the second letter represents sex (F female, M male) and the last 3 digits represent animal serial number.

Dosage of administration: the doses of compound 2, 5, 8 and the comparative compound group were each 20 mg/kg; route of administration: oral administration.

Blood sample collection and detection analysis

(1) PK blood collection

Sampling time for compound 2, 5, 8 and comparative compound group: pre-and post-dose 2 min (+ -1 min), 10min (+ -1 min), 30min (+ -1 min), 1 h (+ -2 min), 2 h (+ -5 min), 4 h (+ -5 min), 6 h (+ -5 min), 8 h (+ -10 min), 10 h (+ -10 min), and 24 h (+ -10 min);

sampling method and sampling amount: jugular vein blood sampling is about 0.15 mL whole blood, sampling is carried outEDTA.K is put in ₂ An anticoagulant tube;

blood sample treatment: placing the whole blood sample in an ice box before centrifugation, transporting the whole blood sample in the ice box, centrifuging for 10min at 2-8 ℃ and 4000 r/min, separating plasma (plasma-1), and storing the whole blood sample below-60 ℃.

(2) Blood concentration detection

The blood concentration of animals in each dose group was measured by using a validated LC-MS/MS method. The concentration-time curves were plotted using Phoenix WinNonlin 7.0.7.0 software and the following pharmacokinetic parameters were calculated in a non-compartmental model: half-life (T) _1/2 ) Maximum blood concentration (C) _max ) Peak time of arrival (T) _max ) Area under the curve of time of Administration (AUC) _last ) Clearance (CL), etc.

BLOQ (below the lower limit of quantitation) was defined before concentration data was imported into WinNonlin 7.0: BLOQ is defined as 0 before there is data, and the remainder as missting.

Data acquisition and analysis

The acquisition system for collecting and reporting electronic data is as follows:

/>

results

(1) Analysis of the formulations for administration

Through detection, the system adaptability, the stock solution comparison and the standard curve linear range and the quality control all meet the requirements of an analysis method. The accuracy (ratio of the detection concentration to the labeling concentration of the administration preparation) of the compound 2, 5 and 8 single groups and the compound of the comparative example single group is 92.92% -102.10%. The analysis results meet the requirements of the test scheme on concentration analysis.

(2) Single administration Pharmacokinetic (PK) results

The average pharmacokinetic parameters for each group after single administration of compounds 2, 5, 8 and the comparative compound are shown in table 7:

。

as can be seen from the above table, the half-lives of Compounds 2, 5, 8 are significantly longer than those of the comparative examples, and AUC, C _max The compound is also superior to the compound of the comparative example, which shows that the deuterated compounds 2, 5 and 8 have slower metabolism in the body and longer action time and have better pharmacokinetic properties, thereby ensuring better pharmacodynamic action.

Claims

1. A compound or a pharmaceutically acceptable salt thereof, wherein the compound is:

2. the compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein the compound is:

3. a compound according to any one of claims 1 to 2, or a pharmaceutically acceptable salt thereof, wherein the salt is an acetate, ascorbate, benzoate, benzenesulfonate, citrate, fumarate, hydrochloride, hydrobromide, maleate, methanesulfonate, nitrate, oxalate, phosphate, succinate or sulfate salt.

4. A pharmaceutical composition comprising a compound according to any one of claims 1 to 2 or a pharmaceutically acceptable salt thereof, wherein the composition further comprises a pharmaceutically acceptable carrier and/or adjuvant.

5. Use of a compound according to any one of claims 1 to 2, or a pharmaceutically acceptable salt thereof, for the manufacture of a medicament for the prophylaxis or treatment of a C-Kit and/or PDGFR mediated disorder.

6. The use according to claim 5, wherein the C-Kit and/or PDGFR mediated disease is selected from gastrointestinal stromal tumor, lupus, leukemia, mastocytosis, melanoma, seminoma, rheumatoid arthritis, multiple sclerosis, cryo-progressive, multiple myeloma or pancreatic cancer.

7. The use according to claim 5, wherein the C-Kit and/or PDGFR is a mutant C-Kit and/or PDGFR.