Deuterated RAS inhibitor drug and application

Abstract

The invention discloses a deuterated RAS inhibitor which is shown in the following formula I. The invention relates to the deuterated RAS inhibitor and a pharmaceutical composition and application thereof. # imgabs0 #

Description

Technical Field

[0001] The present invention belongs to the field of biomedicine, and specifically relates to deuterated RAS inhibitor drugs and uses. Background Art

[0002] Pancreatic cancer is one of the most deadly malignancies and is often diagnosed at an advanced stage. Patients respond poorly to standard chemotherapy and have a very high mortality rate.

[0003] Pancreatic ductal adenocarcinoma (PDAC) is the most common type of pancreatic cancer, accounting for approximately 92% of all pancreatic cancer cases. RAS, the first oncogene discovered in the human body, has long been a hot topic in drug development. There are three RAS genes in the human body: HRAS, KRAS, and NRAS. RAS possesses GTPase activity. When bound to GTP, RAS is active; when bound to GDP, it is inactive. RAS acts as a molecular switch in RAS-mediated cell signaling (RAS / RAF / MEK / ERK and PI3K / AKT signaling pathways), regulating cell differentiation, proliferation, and apoptosis. Over 30% of human tumors are associated with RAS gene activation. RMC-6236 is an oral, multi-targeted RAS inhibitor that exhibits broad anti-tumor activity by inhibiting signaling in the RAS (ON) state, blocking cancer cell proliferation and survival signals.

[0004] Deuterated drugs are new drug molecules that replace one or more carbon-hydrogen bonds in a drug molecule with carbon-deuterium bonds. They can improve the pharmacokinetic properties of the original drugs, thereby overcoming the original defects of the drugs such as easy metabolism and large side effects.

[0005] The present invention is a deuterated RAS inhibitor drug, which can further improve the pharmacokinetic properties of current RAS inhibitor drugs, reduce the dosage and possible toxic side effects. Summary of the Invention

[0006] The present invention provides a deuterated compound of a RAS inhibitor, its stereoisomers, tautomers, and pharmaceutically acceptable salts thereof, which can further improve the pharmacokinetic properties of the deuterated compound of a RAS inhibitor, its stereoisomers, tautomers, and pharmaceutically acceptable salts thereof, and reduce the dosage and possible toxic side effects.

[0007] In order to achieve the above-mentioned object, the present invention provides a deuterated compound of a RAS inhibitor as shown in the following formula I, its stereoisomers, tautomers and pharmaceutically acceptable salts thereof:

[0008]

[0009] The deuterated compounds of the RAS inhibitors of the present invention, their stereoisomers, tautomers, and pharmaceutically acceptable salts thereof, include the following structures:

[0010]

[0011] The deuterated compound of the RAS inhibitor of the present invention, its stereoisomers, tautomers and pharmaceutically acceptable salts thereof, wherein the pharmaceutically acceptable salt is selected from methanesulfonate, maleate, hydrochloride or phosphate.

[0012] The deuterated compound, its stereoisomers, tautomers and pharmaceutically acceptable salts described in the present invention include their use in preparing drugs for preventing and / or treating RAS-mediated diseases.

[0013] RAS inhibitors are provided herein and are known to those skilled in the art. RAS inhibitors are inhibitors that target, i.e., selectively bind to, or inhibit the GDP-bound, inactive state of RAS. Inhibition of the GDP-bound, inactive state of RAS includes, for example, isolating the inactive state by inhibiting the exchange of GDP for GTP, thereby inhibiting RAS from assuming the active conformation. In certain embodiments, RAS inhibitors may also bind to or inhibit the GTP-bound, active state of RAS.

[0014] The deuterated compound, its stereoisomers, tautomers and pharmaceutically acceptable salts thereof described in the present invention comprise the deuterated compound, its stereoisomers, tautomers and pharmaceutically acceptable salts thereof as active ingredients and a pharmaceutically acceptable carrier.

[0015] Certain compounds of the present invention may exist in specific geometric or stereoisomeric forms. The present invention encompasses all compounds, including their cis and trans isomers, R and S enantiomers, diastereomers, (D) isomers, (L) isomers, racemic mixtures, and other mixtures. Additionally, asymmetric carbon atoms may represent substituents, such as alkyl groups. All isomers and mixtures thereof are encompassed by the present invention.

[0016] According to the present invention, mixtures of isomers can contain various ratios of isomers. For example, mixtures containing only two isomers can have the following ratios: 50:50, 60:40, 70:30, 80:20, 90:10, 95:5, 96:4, 97:3, 98:2, 99:1, or 100:0. All ratios of isomers are within the scope of the present invention. Similar ratios, as well as ratios for more complex mixtures of isomers, are readily understood by those skilled in the art and are also within the scope of the present invention.

[0017] The pharmaceutical composition of the deuterated compound and pharmaceutically acceptable salt thereof of the present invention is selected from capsules, powders, tablets, granules, pills, injections, syrups, oral liquids, inhalants, ointments, suppositories or patches.

[0018] Beneficial effects: Compared with the prior art, the present invention has the following advantages:

[0019] (1) The compounds provided by the present invention have excellent KRAS G12C Inhibitory activity;

[0020] (2) The present invention provides a class of deuterated compounds, their stereoisomers, tautomers and pharmaceutically acceptable salts thereof, which further improve the pharmacokinetic properties of RAS inhibitors, reduce the dosage and possible toxic side effects. BRIEF DESCRIPTION OF THE DRAWINGS

[0021] In order to more clearly illustrate the embodiments of the present invention and the technical solutions of the prior art, the following briefly introduces the drawings required for use in the embodiments and the prior art. Obviously, the drawings described below are only some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying any creative work.

[0022] Figure 1 This is the hydrogen spectrum characterization spectrum of the compound in Example 1. DETAILED DESCRIPTION

[0023] The present invention will be further described below with reference to the following examples. The present invention will be further described below with reference to the following specific examples. It should be understood that these examples are intended only to illustrate the present invention and are not intended to limit the scope of the present invention.

[0024] Unless otherwise defined, all technical and scientific terms used herein have the same meanings as those familiar to those skilled in the art. Furthermore, any methods and materials similar or equivalent to those described herein can be applied to the methods of the present invention. The preferred embodiments and materials described herein are for illustrative purposes only. Specific embodiments

[0026] Example 1

[0027] Example 1 General Synthesis Method of Compounds

[0028]

[0029] The synthesis of intermediate 1-E can be prepared by referring to the synthesis method of intermediate 4 in patent CN116457358.

[0030] The synthesis of intermediate 1-I can be prepared by referring to the synthesis method of intermediate 5 in patent CN116457358.

[0031] Synthesis of intermediate 1-J

[0032] To a THF solution (10 mL) of Intermediate 1-I (0.82 g, 1 mmol) was added sodium hydroxide (0.1 g, 4 mmol, 4 eq) at 0°C. The mixture was allowed to react at room temperature for half an hour. Deuterated iodomethane was then added and the reaction continued for 2 hours. TLC confirmed the reaction was complete and the reaction was quenched by adding water. The aqueous phase was extracted with ethyl acetate (10 mL x 2), washed with water (20 mL x 2), washed with saturated sodium chloride (20 mL), dried over anhydrous sodium sulfate, and concentrated by column chromatography to yield Intermediate 1-J (577 mg, 69%). m / z (ESI, MS): 833.10.

[0033] Synthesis of compounds

[0034] The compound of Example 1 (344 mg, 53%) was prepared by referring to the synthesis method of Example A120 in patent CN116457358. 1 H NMR (500MHz, DMSO-d6) δ8.53(d,J=4.8Hz,1H),8.47(dd,J=6.2,4.2Hz,2H),7.80(s,1H),7.73(dd,J=4.0,2.2Hz,1H),7.56( d,J=5.0Hz,1H),7.22(d,J=2.5Hz,1H),5.56(t,J=2.5Hz,1H),5.10(d,J=7.5Hz,1H),4.26-4.14(m,5H),3.57(s,2H),3.28- 3.25(m,4H),3.24(s,3H),3.21-3.15(m,1H),2.94(d,J=9.0Hz,1H),2.78-2.73(m,1H),2.52-3.47(m,4H),2.10-2.06(m,1H ),1.79(s,2H),1.51-1.49(m,2H),1.34-1.32(m,3H),1.08-1.07(m,4H),0.91-0.87(m,7H),0.57-0.54(m,1H),0.34(s,3H).

[0035] Example 2

[0036] Example 2 General Synthesis Method of Compounds

[0037]

[0038] Synthesis of intermediate 2-A

[0039] To a THF solution (10 mL) of Intermediate 1-A (0.2 g, 1 mmol) at 0°C was added sodium hydroxide (0.1 g, 4 mmol, 4 eq). The mixture was allowed to react at room temperature for half an hour. Deuterated iodomethane was then added and the reaction continued for 2 hours. TLC confirmed the reaction was complete and the reaction was quenched by adding water. The aqueous phase was extracted with ethyl acetate (10 mL x 2), washed with water (20 mL x 2), washed with saturated sodium chloride (20 mL), dried over anhydrous sodium sulfate, and concentrated by column chromatography to yield Intermediate 2-A (178 mg, 82%). m / z (ESI, MS): 219.30.

[0040] Synthesis of compounds

[0041] Referring to the synthesis method of Example 1, the intermediate 1-B was replaced by 2-A, and the deuterated iodomethane in the synthesis method of intermediate 1-J was replaced by iodomethane to obtain the compound of Example 2 (321 mg, 51%). 1 H NMR (500MHz, DMSO-d6) δ8.50(d,J=4.5Hz,1H),8.45(dd,J=6.2,4.2Hz,2H),7.81(s,1H),7.75(dd,J=4.0,2.1Hz,1H),7.51( d,J=5.0Hz,1H),7.23(d,J=2.5Hz,1H),5.52(t,J=2.5Hz,1H),5.12(d,J=7.5Hz,1H),4.23-4.12(m,5H),3.52(s,2H),3.28- 3.24(m,4H),3.20-3.15(m,1H),2.93(d,J=9.0Hz,1H),2.72-2.70(m,1H),2.52-3.47(m,4H),2.22(s,3H),2.10-2.04(m,1H ),1.78(s,2H),1.50-1.49(m,2H),1.34-1.31(m,3H),1.08-1.07(m,4H),0.92-0.87(m,7H),0.57-0.54(m,1H),0.33(s,3H).

[0042] Example 3

[0043] Synthesis of compounds

[0044] Referring to the synthetic method of Example 1, the intermediate 1-B was replaced with 2-A to obtain the compound of Example 3 (303 mg, 49%). 1H NMR (500MHz, DMSO-d6) δ8.52(d,J=4.8Hz,1H),8.46(dd,J=6.2,4.2Hz,2H),7.81(s,1H),7.72(d,J=4.0Hz,1H),7.53(d ,J=5.0Hz,1H),7.24(d,J=2.5Hz,1H),5.53(t,J=2.5Hz,1H),5.09(d,J=7.5Hz,1H),4.26-4.15(m,5H),3.56(s,2H),3. 29-3.25(m,4H),3.21-3.16(m,1H),2.95(d,J=9.0Hz,1H),2.78-2.72(m,1H),2.53-3.47(m,4H),2.11-2.06(m,1H),1. 79(s,2H),1.52-1.49(m,2H),1.33-1.32(m,3H),1.10-1.07(m,4H),0.91-0.88(m,7H),0.57-0.54(m,1H),0.34(s,3H).

[0045] Test Example 1: KRAS G12C Inhibitory activity test

[0046] (1) Reagents and consumables

[0047] Reagent name Supplier Item No. <![CDATA[KRAS G12C ]]> Cisbio 63ADK000CB16PEG 384-well plate Perkin Elmer 6007290

[0048] (2) Instruments

[0049] Centrifuge (manufacturer: Eppendorf, model: 5430 )

[0050] Microplate reader (Manufacturer: Perkin Elmer, Model: EnVision )

[0051] (3) Experimental methods

[0052] Test compounds were prepared in DMSO, diluted proportionally, and added to 384-well plates. A Tag1-SOS1 solution was prepared in Diluent Buffer at a 4x final concentration. 2.5 μL of this solution was added to each well. A Tag2-KRAS-G12C solution was also added in Diluent Buffer. The 384-well plate was centrifuged at 1000 rpm for 30 seconds, shaken thoroughly, and incubated at room temperature for 15 minutes. Anti-Tag1-TB3+ and Anti-Tag2-XL665 solutions were prepared in Detection Buffer at 1x final concentrations, mixed thoroughly, and 5 μL was added to each well. The 384-well plate was centrifuged at 1000 rpm for 30 seconds, shaken thoroughly, and incubated at room temperature for 2 hours. The concentrations were read using a microplate reader (Em665 / 620).

[0053] Compound <![CDATA[KRAS G12C IC 50 (nM)]]> Example 1 0.8 Example 2 1.1 Example 3 0.7 RMC-6236 1.8

[0054] The above table shows that Example compounds 1-3 all exhibit excellent KRAS G12C In terms of inhibitory activity, compared to the positive drug RMC-6236, the activity of the compounds in Example 1 and Example 3 was increased by two times, and the activity gain was significant.

[0055] (1) Experimental instruments and materials

[0056] High-speed refrigerated centrifuges, vortex shakers (Vortex Genius 3), high-speed centrifuges (Eppendorf 5415D), disposable syringes, pipettes (Eppendorf), and SD male rats used in the experiment were purchased from Yangzhou University. EDTA-K2 vacuum blood collection tubes and normal saline were used. All rats in the oral groups fasted for 12 hours before administration and had free access to water and food during the administration period.

[0057] (2) Experimental steps

[0058] Example 1 or RMC-6236 was dissolved in DMSO / solutol / water (10 / 10 / 80) to prepare a clear solution. The compound was administered orally at a dose of 25 mg / kg, and the compound was administered via tail vein at a dose of 5 mg / kg. Continuous blood samples (0.5 mL) were collected from the retinal venous plexus into heparinized tubes at 2, 10, 30, 1, 2, 3, 5, 8, 12, 16, and 24 hours after tail vein administration. Continuous blood samples (0.5 mL) were collected from the retinal venous plexus into heparinized tubes at 5, 15, 30, 1, 2, 3, 5, 8, 12, 16, and 24 hours after gavage administration. After centrifugation at 8000 r / min at 4°C for 10 minutes, 0.15 mL of the upper plasma layer was collected and stored at -20°C prior to LC-MS / MS analysis. The data were analyzed using a WinNolin non-compartmental model to determine key pharmacokinetic parameters.

[0059] (3) Experimental results

[0060] Table 1 Pharmacokinetic parameters

[0061]

[0062]

[0063] Compared with the positive drug RMC-6236, the oral half-life of Example 3 is doubled, and the peak concentration is also significantly increased, which can effectively improve the dosage of RMC-6236, thereby reducing the toxic side effects of high-dose administration of RMC-6236.

[0064] Finally, it should be noted that the above content is only used to illustrate the technical solution of the present invention, rather than to limit the scope of protection of the present invention. Simple modifications or equivalent substitutions of the technical solution of the present invention by ordinary technicians in this field do not deviate from the essence and scope of the technical solution of the present invention.

Claims

1. A deuterated RAS inhibitor compound of formula I, its stereoisomers, tautomers and pharmaceutically acceptable salts thereof, in, R1 and R2 are independently selected from hydrogen or deuterium, and R1 and R2 are not hydrogen.

2. The deuterated compound, its stereoisomers, tautomers and pharmaceutically acceptable salts according to claim 1, characterized in that: The compound is selected from the following structures:

3. The deuterated compound, stereoisomers, tautomers and pharmaceutically acceptable salts thereof according to claim 1, characterized in that The pharmaceutically acceptable salt is selected from methanesulfonate, maleate, hydrochloride or phosphate.

4. Use of the deuterated compound according to claim 1, its stereoisomers, tautomers and pharmaceutically acceptable salts thereof in the preparation of a medicament for preventing and / or treating RAS-mediated diseases.

5. A pharmaceutical composition, characterized in that The pharmaceutical composition is composed of the deuterated compound according to claim 1, its stereoisomers, tautomers and pharmaceutically acceptable salts thereof as active ingredients and a pharmaceutically acceptable carrier.

6. The pharmaceutical composition according to claim 5, characterized in that The pharmaceutical composition is selected from capsules, powders, tablets, granules, pills, injections, syrups, oral solutions, inhalants, ointments, suppositories or patches.

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