Phosphate prodrug of Malt1 inhibitor

Abstract

The present disclosure provides a phosphate prodrug of a Malt1 inhibitor. Specifically, the invention provides a prodrug as shown in a formula (I) or a pharmaceutically acceptable salt thereof and application thereof, and the prodrug or the pharmaceutically acceptable salt can be used for preparing drugs. # imgabs0 #

Description

A phosphate prodrug of a Malt 1 inhibitor

[0001] This application claims priority to Chinese patent application No. 2022116209806, filed on December 16, 2022. This application incorporates the entirety of the aforementioned Chinese patent application. Technical Field

[0002] The present invention relates to a compound represented by formula I and a preparation method and use thereof, and belongs to the field of medicine. Background Art

[0003] Mucosa-associated-lymphoid-tissue lymphoma-translocation 1 (MALT1) is a key upstream protein in the NF-κB signaling pathway. It forms a complex (CBM) with B-cell chronic lymphocytic leukemia / lymphoma 10 (BCL10) and caspase-recruitment domain (CARD)-containing membrane-associated guanylate kinase protein 1 (CARMA1). MALT1 transmits signals from proximal antigen receptor proteins to IκB kinase (IKK), thereby activating the NF-κB signaling pathway. Overactivation of the MALT1-NF-κB signaling pathway is closely associated with inflammation and tumorigenesis.

[0004] Patent application PCT / CN2022 / 099499 discloses a class of Malt 1 inhibitors, and the present disclosure incorporates the entire contents of patent application PCT / CN2022 / 099499 into the present disclosure.

[0005] Summary of the Invention

[0006] In a first aspect, the present disclosure provides a prodrug of a compound represented by Formula I or a pharmaceutically acceptable salt thereof,

[0007] In a second aspect, the present disclosure further provides a prodrug of the compound of Formula I, which can be selected from a phosphate prodrug of the compound of Formula I, a dicarboxylic acid ester prodrug of the compound of Formula I, or an amino acid ester prodrug.

[0008] In some embodiments, the prodrug of the compound of formula I as described in the first aspect can be a phosphate prodrug of the compound of formula I or a pharmaceutically acceptable salt thereof.

[0009] In some embodiments, the phosphate prodrug of the compound of formula I may be a compound structure shown in formula II-a,

[0010] n is selected from integers of 1-5, preferably n is selected from integers of 1-3, and more preferably n is selected from integers of 1-2.

[0011] In some embodiments, the prodrug of the compound of formula I as described in the first aspect can be a dicarboxylic acid ester prodrug of the compound of formula I.

[0012] In certain embodiments, the dicarboxylic acid ester prodrug of the compound of formula I is a structure shown in formula II-b

[0013] m is selected from integers of 1-5, preferably m is selected from integers of 1-3, and more preferably m is selected from integers of 1-2.

[0014] In some embodiments, the prodrug of the compound of formula I as described in the first aspect can be an amino acid ester prodrug of the compound of formula I, wherein the amino acid can be a common amino acid structure.

[0015] The amino acids are preferably naturally occurring amino acids. More preferably, the amino acids are α-amino acids. Also preferably, the amino acids are L-form. Preferred amino acids include the twenty essential amino acids. Preferred amino acids are lysine (Lys), leucine (Leu), isoleucine (Ile), glycine (Gly), aspartic acid (Asp), glutamic acid (Glu), methionine (Met), alanine (Ala), valine (Val), proline (Pro), histidine (His), tyrosine (Tyr), serine (Ser), norleucine (Nor), arginine (Arg), phenylalanine (Phe), tryptophan (Trp), hydroxyproline (Hyp), homoserine (Hsr), carnitine (Car), ornithine (Ort), canavanine (Cav), asparagine (Asn), glutamine (Gln), carnosine (Can), taurine (Tau), S-methylenecystine (Djk), gamma-aminobutyric acid (GABA), cysteine ​​(Cys), cystine (Dcy), sarcosine (Sar), threonine (Thr), and the like. More preferred amino acids are the twenty essential amino acids, Lys, Leu, Ile, Gly, Asp, Glu, Met, Ala, Val, Pro, His, Tyr, Thr, Arg, Phe, Trp, Gln, Asn, Cys, and Ser.

[0016] In certain embodiments, the amino acid ester prodrug of the compound of formula I is a structure shown in formula II-c

[0017] R is a side group or side chain of an amino acid.

[0018] In a third aspect, the present disclosure further provides a compound as shown in Formula III or a pharmaceutically acceptable salt thereof,

[0019] In some embodiments, the compound or pharmaceutically acceptable salt thereof as described in the first, second and third aspects, wherein the pharmaceutically acceptable salt can be selected from inorganic salts or organic salts, and may also include pharmaceutically acceptable acid addition salts and pharmaceutically acceptable base addition salts.

[0020] "Pharmaceutically acceptable acid addition salts" refer to salts formed with inorganic or organic acids that retain the biological effectiveness of the free base without other side effects. Inorganic acid salts include, but are not limited to, hydrochlorides, hydrobromides, sulfates, nitrates, and phosphates; organic acid salts include, but are not limited to, formates, acetates, 2,2-dichloroacetates, trifluoroacetates, propionates, caproates, caprylates, decanoates, undecylenates, glycolates, gluconates, lactates, sebacates, adipates, glutarates, malonates, oxalates, maleates, succinates, fumarates, tartrates, citrates, palmitates, stearates, oleates, cinnamates, laurates, malates, glutamate, pyroglutamate, aspartate, benzoates, methanesulfonates, benzenesulfonates, p-toluenesulfonates, alginate, ascorbate, salicylates, 4-aminosalicylates, and naphthalene disulfonates. These salts can be prepared by methods known in the art.

[0021] "Pharmaceutically acceptable base addition salts" refer to salts formed with inorganic or organic bases that retain the biological effectiveness of the free acid without other side effects. Salts derived from inorganic bases include, but are not limited to, sodium salts, potassium salts, lithium salts, ammonium salts, calcium salts, magnesium salts, iron salts, zinc salts, copper salts, manganese salts, aluminum salts, and the like. Preferred inorganic salts are ammonium salts, sodium salts, potassium salts, calcium salts, and magnesium salts, with sodium salts being preferred. The salt derived from organic base includes but is not limited to following salt: primary amines, secondary amines and tertiary amines, substituted amines, including natural substituted amines, cyclic amines and basic ion exchange resins, such as ammonia, isopropylamine, trimethylamine, diethylamine, triethylamine, tripropylamine, ethanolamine, diethanolamine, triethanolamine, dimethylethanolamine, 2-dimethylaminoethanol, 2-diethylaminoethanol, dicyclohexylamine, lysine, arginine, histidine, caffeine, procaine, choline, betaine, ethylenediamine, glucosamine, methylglucamine, theobromine, purine, piperazine, piperidine, N-ethylpiperidine, polyamine resins etc. Preferred organic bases include isopropylamine, diethylamine, ethanolamine, trimethylamine, dicyclohexylamine, choline and caffeine. These salts can be prepared by methods known in the art.

[0022] In some embodiments, the compound or pharmaceutically acceptable salt thereof as described in the first, second, and third aspects, wherein the pharmaceutically acceptable salt is a sodium salt.

[0023] In a fourth aspect, the present disclosure further provides a compound represented by formula IV or a pharmaceutically acceptable salt thereof,

[0024] In a fifth aspect, the present disclosure provides an isotope-substituted prodrug or a pharmaceutically acceptable salt thereof according to any one of the first to fourth aspects, preferably, the isotope substitution is deuterium atom substitution.

[0025] In a sixth aspect, the present disclosure provides a pharmaceutical composition comprising the prodrug or a pharmaceutically acceptable salt thereof as described in any one of the first to fourth aspects or the isotope substitution as described in the fifth aspect and a pharmaceutically acceptable excipient.

[0026] In some embodiments, the unit dose of the pharmaceutical composition is 0.001 mg-1000 mg.

[0027] In certain embodiments, the pharmaceutical composition contains 0.01-99.99% of the aforementioned compound or a pharmaceutically acceptable salt thereof, based on the total weight of the composition. In certain embodiments, the pharmaceutical composition contains 0.1-99.9% of the aforementioned compound or a pharmaceutically acceptable salt thereof. In certain embodiments, the pharmaceutical composition contains 0.5%-99.5% of the aforementioned compound or a pharmaceutically acceptable salt thereof. In certain embodiments, the pharmaceutical composition contains 1%-99% of the aforementioned compound or a pharmaceutically acceptable salt thereof. In certain embodiments, the pharmaceutical composition contains 2%-98% of the aforementioned compound or a pharmaceutically acceptable salt thereof.

[0028] In certain embodiments, the pharmaceutical composition comprises 0.01% to 99.99% of a pharmaceutically acceptable excipient, based on the total weight of the composition. In certain embodiments, the pharmaceutical composition comprises 0.1% to 99.9% of a pharmaceutically acceptable excipient. In certain embodiments, the pharmaceutical composition comprises 0.5% to 99.5% of a pharmaceutically acceptable excipient. In certain embodiments, the pharmaceutical composition comprises 1% to 99% of a pharmaceutically acceptable excipient. In certain embodiments, the pharmaceutical composition comprises 2% to 98% of a pharmaceutically acceptable excipient.

[0029] The present disclosure also provides use of a prodrug according to any one of the first to fourth aspects or a pharmaceutically acceptable salt thereof, or the isotope substitution according to the fifth aspect, or the pharmaceutical composition according to the sixth aspect in the preparation of a medicament for preventing and / or treating a disorder associated with MALT1.

[0030] The present disclosure also provides a prodrug according to any one of the first to fourth aspects or a pharmaceutically acceptable salt thereof, or the isotope substitution according to the fifth aspect, or a pharmaceutical composition according to the sixth aspect for preventing and / or treating a disorder associated with MALT1.

[0031] The present disclosure also provides use of a prodrug or a pharmaceutically acceptable salt thereof as described in any one of the first to fourth aspects, or the isotope substitution according to the fifth aspect, or the pharmaceutical composition according to the sixth aspect in the preparation of a medicament for preventing and / or treating autoimmune diseases, inflammatory diseases, cancer, and tumors.

[0032] The present disclosure also provides a prodrug or a pharmaceutically acceptable salt thereof as described in any one of the first to fourth aspects, or the isotope substitution according to the fifth aspect, or the pharmaceutical composition according to the sixth aspect for preventing and / or treating autoimmune diseases, inflammatory diseases, cancer, and tumors.

[0033] The autoimmune diseases and inflammatory diseases, such as rheumatoid arthritis, multiple sclerosis, systemic lupus erythematosus or vasculitis, the cancer or tumor, such as primary cancer of the hematopoietic system or solid tumors, including chronic myeloid leukemia, myeloid leukemia, non-Hodgkin's lymphoma and other B-cell lymphomas.

[0034] The present disclosure provides a prodrug of a compound represented by Formula I or a pharmaceutically acceptable salt thereof, wherein the compound of Formula I has good Malt1 inhibitory activity and, when prepared as a prodrug, can significantly improve water solubility and pK effect.

[0035] Explanation of terms:

[0036] A "pharmaceutical composition" refers to a mixture containing one or more compounds described herein, or their physiologically acceptable salts or prodrugs, together with other chemical components, as well as other components such as physiologically acceptable carriers and excipients. The purpose of a pharmaceutical composition is to facilitate administration to an organism, facilitating absorption of the active ingredients and thereby exerting their biological activity.

[0037] "Pharmaceutically acceptable excipients" include, but are not limited to, any adjuvant, carrier, excipient, glidant, sweetener, diluent, preservative, dye / colorant, flavoring agent, surfactant, wetting agent, dispersant, suspending agent, stabilizer, isotonic agent, solvent or emulsifier approved by the U.S. Food and Drug Administration for use in humans or domestic animals.

[0038] As used herein, an "effective amount" or "therapeutically effective amount" encompasses an amount sufficient to ameliorate or prevent the symptoms or conditions of a medical condition. An effective amount also refers to an amount sufficient to permit or facilitate diagnosis. The effective amount for a particular patient or veterinary subject may vary depending on factors such as the condition to be treated, the patient's overall health, the method, route, and dosage of administration, and the severity of side effects. An effective amount can be the maximum dose or dosage regimen that avoids significant side effects or toxic effects. DETAILED DESCRIPTION

[0039] The present invention is further described below with reference to the following examples, but these examples are not intended to limit the scope of the present invention.

[0040] Example

[0041] The structures of the compounds were determined by nuclear magnetic resonance (NMR) and / or mass spectrometry (MS). -6 The unit of ppm is given. NMR measurements were performed using a Bruker AVANCE-400 NMR spectrometer. The solvents used were deuterated dimethyl sulfoxide (DMSO-d6), deuterated chloroform (CDCl3), and deuterated methanol (CD3OD), and tetramethylsilane (TMS) was used as the internal standard.

[0042] MS was measured using a Shimadzu 2010 Mass Spectrometer or an Agilent 6110A MSD mass spectrometer.

[0043] High-performance liquid chromatography (HPLC) was performed using a Shimadzu LC-20A systems, Shimadzu LC-2010HT series, Shimadzu DGU-20A5R, Shimadzu LC-30AD, Shimadzu SIL-30AC, or Agilent 1200 LC high-pressure liquid chromatograph (Ultimate XB-C18 3.0*150 mm column or Xtimate C18 2.1*30 mm column).

[0044] Chiral HPLC analysis was performed using Chiralpak IC-3 100×4.6mm ID, 3um, Chiralpak AD-3 150×4.6mm ID, 3um, Chiralpak AD-3 50×4.6mm ID, 3um, Chiralpak AS-3 150×4.6mm ID, 3um, Chiralpak AS-3 100×4.6mm ID, 3μm, ChiralCel OD-3 150×4.6mm ID, 3um, Chiralcel OD-3 100×4.6mm ID, 3μm, ChiralCel OJ-H 150×4.6mm ID, 5um, and Chiralcel OJ-3 150×4.6mm ID, 3um columns.

[0045] The thin layer chromatography silica gel plate uses Yantai Huanghai HSGF254 or Qingdao GF254 silica gel plate. The specification of the silica gel plate used in thin layer chromatography (TLC) is 0.15mm~0.2mm, and the specification used for thin layer chromatography separation and purification products is 0.4mm~0.5mm.

[0046] Column chromatography generally uses Yantai Huanghai silica gel 100-200 mesh, 200-300 mesh or 300-400 mesh silica gel as the carrier.

[0047] The chiral preparative column used was DAICEL CHIRALPAK IC (250 mm*30 mm, 10 um) or Phenomenex-Amylose-1 (250 mm*30 mm, 5 um).

[0048] The CombiFlash rapid preparation instrument used was Combiflash Rf150 (TELEDYNE ISCO).

[0049] Average kinase inhibition rate and IC 50 The values ​​were determined using a NovoStar microplate reader (BMG, Germany).

[0050] The known starting materials of the present invention can be synthesized by methods known in the art, or can be purchased from companies such as ABCR GmbH & Co. KG, Acros Organics, Aldrich Chemical Company, Accela ChemBio Inc, and Darui Chemicals.

[0051] Unless otherwise specified in the examples, all reactions can be carried out under an argon atmosphere or a nitrogen atmosphere.

[0052] Argon atmosphere or nitrogen atmosphere means that the reaction bottle is connected to an argon or nitrogen balloon with a capacity of about 1 L.

[0053] Hydrogen atmosphere means that the reaction bottle is connected to a hydrogen balloon with a capacity of about 1L.

[0054] The pressurized hydrogenation reaction uses a Parr 3916EKX hydrogenator and a Qinglan QL-500 hydrogen generator or an HC2-SS hydrogenator.

[0055] The hydrogenation reaction is usually carried out by evacuating the chamber and filling it with hydrogen, and the operation is repeated three times.

[0056] A CEM Discover-S 908860 microwave reactor was used for the microwave reaction.

[0057] Unless otherwise specified in the examples, the solution refers to an aqueous solution.

[0058] Unless otherwise specified in the examples, the reaction temperature is room temperature, 20°C to 30°C.

[0059] The reaction progress in the examples was monitored by thin layer chromatography (TLC). The developing solvent used in the reaction, the column chromatography eluent system used for purifying the compound, and the developing solvent system for thin layer chromatography, the volume ratio of the solvent were adjusted according to the polarity of the compound. Generally, water and acetonitrile were used as the mobile phase, and a small amount of alkaline or acidic reagents such as triethylamine and acetic acid could be added for adjustment.

[0060] Example 1

[0061] N-(Benzo[c][1,2,5]oxadiazol-5-yl)-1-(1-oxo-1,2-dihydroisoquinolin-5-yl)-5-(trifluoromethyl)-1H-pyrazole-4-carboxamide 1

[0062] first step

[0063] 5-Hydrazinoisoquinoline 1b

[0064] Isoquinolin-5-amine (5.1 g, 35.37 mmol) was dissolved in concentrated hydrochloric acid (50 mL). A solution of sodium nitrite (3.66 g, 53.06 mmol) in water (20 mL) was added at 0°C. After 30 minutes of reaction, a solution of stannous chloride (19.95 g, 88.43 mmol) in concentrated hydrochloric acid (20 mL) was added dropwise. The reaction was continued at room temperature for 3 hours. The pH was adjusted to 12-14 with 20% aqueous sodium hydroxide solution. The product was extracted with ethyl acetate (30 mL*3). The organic phase was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography eluting with ethyl acetate to give the title compound 1b (2.33 g, yield: 40.0%).

[0065] MS (ESI): m / z = 159.0 [M+H] + .

[0066] Step 2

[0067] 1-(Isoquinolin-5-yl)-5-(trifluoromethyl)-1H-pyrazole-4-carboxylic acid ethyl ester 1d

[0068] 5-Hydrazinoisoquinoline (2.33 g, 14.64 mmol) and (Z)-2-(ethoxymethylene)-4,4,4-trifluoro-3-oxobutanoic acid ethyl ester 1c (3.52 g, 14.64 mmol) were dissolved in ethanol (40 mL) and reacted at 60°C for 3 hours. The mixture was concentrated under reduced pressure to give a crude product, which was purified by silica gel column chromatography eluting with ethyl acetate to give the title compound 1d (2.56 g, yield: 52.1%).

[0069] MS (ESI): m / z = 336.4 [M+H] + .

[0070] Step 3

[0071] 5-(4-(Ethoxycarbonyl)-5-(trifluoromethyl)-1H-pyrazol-1-yl)isoquinoline 2-oxide 1e

[0072] Dissolve 1-(isoquinolin-5-yl)-5-(trifluoromethyl)-1H-pyrazole-4-carboxylic acid ethyl ester (2.56 g, 7.64 mmol) in dichloromethane (30 mL), add m-chloroperbenzoic acid (3.95 g, 22.9 mmol) at 0°C, react at room temperature overnight, wash with half-saturated sodium bisulfite solution (20 mL*2) and potassium carbonate solution (20 mL*2) in sequence, dry the organic phase, and concentrate in vacuo to give the title compound 1e (2.50 g, yield: 93.21%).

[0073] MS (ESI): m / z = 352.4 [M+H] + .

[0074] Step 4

[0075] 1-(1-chloroisoquinolin-5-yl)-5-(trifluoromethyl)-1H-pyrazole-4-carboxylic acid ethyl ester 1f

[0076] 5-(4-(Ethoxycarbonyl)-5-(trifluoromethyl)-1H-pyrazol-1-yl)isoquinoline 2-oxide (2.5 g, 7.12 mmol) was dissolved in chloroform (30 mL), and phosphorus oxychloride (1.33 mL, 14.23 mmol) was added at room temperature. The reaction was carried out at 60°C for 3 hours, and the reaction was quenched with water (20 mL) in an ice bath. The organic phase was concentrated in vacuo and purified by silica gel column chromatography eluting with petroleum ether and ethyl acetate to give the title compound 1f (1.96 g, yield: 74.49%).

[0077] MS (ESI): m / z = 370.43 [M+H] + .

[0078] Step 5

[0079] 1-(1-Oxo-1,2-dihydroisoquinolin-5-yl)-5-(trifluoromethyl)-1H-pyrazole-4-carboxylic acid 1 g

[0080] Ethyl 1-(1-chloroisoquinolin-5-yl)-5-(trifluoromethyl)-1H-pyrazole-4-carboxylate (1 g, 2.71 mmol) was dissolved in concentrated hydrochloric acid (15 mL), reacted at 120°C for 3 hours, and concentrated in vacuo to give 1 g (864 mg, yield: 98.83%) of the title compound.

[0081] MS (ESI): m / z = 324.4 [M+H] + .

[0082] Step 6

[0083] Benzo[c][1,2,5]oxadiazole-5-amine 1i

[0084] 5-Bromobenzo[c][1,2,5]oxadiazole 1h (250 mg, 1.25 mmol) was dissolved in aqueous ammonia (1.5 mL) and N-methylpyrrolidone (1 mL), and cuprous oxide (36 mg, 0.25 mmol) was added. The reaction was carried out in a microwave oven at 140°C for 1 hour. After the reaction was completed, 10 ml of ethyl acetate was added, and the mixture was washed twice with 10 ml of water and 10 ml of saturated NaCl aqueous solution. The organic phase was concentrated to obtain the title compound 1i (180 mg, crude product), which was used directly in the next step without further purification.

[0085] MS (ESI) m / z: 136.3 [M+H] + .

[0086] Step 7

[0087] N-(Benzo[c][1,2,5]oxadiazol-5-yl)-1-(1-oxo-1,2-dihydroisoquinolin-5-yl)-5-(trifluoromethyl)-1H-pyrazole-4-carboxamide 1

[0088] Benzo[c][1,2,5]oxadiazole-5-amine 1i (50 mg, 0.37 mmol) was dissolved in pyridine (3 mL), and 1-(1-oxo-1,2-dihydroisoquinolin-5-yl)-5-(trifluoromethyl)-1H-pyrazole-4-carboxylic acid 1 g (119 mg, 0.37 mmol) was added. Phosphorus oxychloride (113 mg, 0.74 mmol) was added at room temperature, and the reaction was carried out at room temperature for 2 hours. The reaction solution was concentrated under reduced pressure, purified by C18 reverse phase column chromatography, and lyophilized to give the title compound 1 (8.3 mg, yield: 5%).

[0089] MS (ESI): m / z = 441.6 [M+H] + .

[0090] 1 H NMR(400MHz, DMSO-d6)δ11.65(br s, 1H), 11.13 (s, 1H), 8.54 (s, 2H), 8.45 (d, J=8.0Hz, 1H), 8.13 (d, J=9.6Hz, 1H), 7.95 (d, J=7.2Hz, 1H), 7.76-7.64 (m, 2H), 7.31 (br s, 1H), 5.67 (d, J=7.6Hz, 1H).

[0091] Example 2

[0092] Sodium (5-(4-(benzo[c][1,2,5]oxadiazol-5-ylcarbamoyl)-5-(trifluoromethyl)-1H-pyrazol-1-yl)-1-oxoisoquinolin-2(1H)-yl)methyl phosphate

[0093] first step

[0094] At room temperature, 2a (148.4 g, 0.337 mol) and paraformaldehyde (151.7 g, 5.06 mol) were suspended in 2000 mL of tetrahydrofuran and heated to 65°C for 1 to 2 hours. After completion of the reaction, the paraformaldehyde was removed by filtration and washed with 6000 mL of tetrahydrofuran. The filtrate was collected, and tetrabenzyl pyrophosphate (236 g, 0.438 mol) was added to the filtrate. The gas was replaced with argon. The reaction system was cooled to 0-5°C, and LiHMDS (506 mL, 0.506 mol) was slowly added dropwise to the reaction system. After the addition was complete, stirring was continued for 30 minutes. The insoluble matter in the reaction system was removed by filtration, and the filtrate was concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography using an eluent system (dichloromethane / methanol = 50 / 1) to obtain the title product 2b (194.5 g, yield: 79%).

[0095] MS m / z(ESI):731[M+1] +

[0096] 1 H NMR (400MHz, d6-DMSO): δ11.11 (s, 1H), 8.56 (d, 1H, J = 10.8Hz), 8.51 (d, 1H, J = 8.0Hz), 8.14 (d, 1H, J = 9.6Hz), 8.03 (d, 1H, J = 7.6Hz) , 7.78-7.72 (m, 2H), 7.62 (d, 1H, J=7.6Hz), 7.32 (s, 10H), 5.91 (dd, 2H, J=9.6, 4.8Hz), 5.81 (d, 1H, J=8.0Hz), 5.06 (d, 4H, J=8.0Hz).

[0097] Step 2

[0098] To a reaction flask, add 2b (193 g, 0.264 mol), 10% Pd / C (19.3 g), and 1500 mL of tetrahydrofuran. Replace the atmosphere with argon. Stir at room temperature until complete conversion of the starting material is achieved. After completion of the reaction, filter through celite to remove the Pd / C. Wash the Pd / C with tetrahydrofuran to keep it moist. Collect the filtrate and concentrate under reduced pressure. After concentration to dryness, add 1000 mL of methanol, filter out a small amount of insoluble impurities, and wash with 500 mL of methanol. Dissolve NaOH (18.9 g, 0.472 mol) in 70 mL of purified water and slowly add dropwise to the methanol solution under an ice bath. After addition, concentrate the methanol solution under reduced pressure to dryness, reconstitute with water, and purify by reverse-phase column chromatography (filler: C18, mobile phase: 10% methanol, 90% water, isocratic elution). Concentrate the prepared solution and lyophilize to obtain the title compound 2 (86 g, yield: 54.7%).

[0099] MS m / z(ESI):551[M+1] +

[0100] 1 H NMR (400MHz, D2O): δ 8.44 (d, 1H, J = 8.0Hz), 8.24 (s, 1H), 8.04 (s, 1H), 7.84 (d, 1H, J = 7.6Hz), 7.76 (d, 1H, J = 9.6Hz), 7. 63 (t, 1H, J = 8.0Hz), 7.56 (d, 1H, J = 7.6Hz), 7.41 (d, 1H, J = 9.6Hz), 6.10 (d, 1H, J = 7.6Hz), 5.59 (dd, 1H, J = 17.4, 6.2Hz).

[0101] Test Example 1: MALT1 biochemical protease assay

[0102] MALT1 protease activity was assessed in an in vitro assay using a tetrapeptide (Ac-LRSR-MCA, PEPTIDE INSTITUTE) as a substrate and full-length MALT1 protein purified from mammalian HEK293T cells (Strep-MALT1(1-824)-Myc / DDK, ORIGENE TP314639). The tetrapeptide LRSR was coupled to AMC (7-amino-4-methylcoumarin) and provided a quenched fluorescent substrate for MALT1 protease. Cleavage of AMC from arginine residues resulted in an increase in coumarin fluorescence measured at 450 nm (excitation 360 nm). The final assay buffer composition was 5.625 nM MALT1 protein, 2.5 μM Ac-LRSR-MCA, 20 mM HEPES, 10 mM KCl, 1.5 mM MgCl·6H2O, 1 mM 2Na (EDTA·2Na), 0.01% Triton X-100, 1 M Trisodium Citrate Dihydrate, and 10 mM DTT. Test compounds dissolved in 100% DMSO were added to a 384-well plate (Greiner-781086) at a volume of 200 nL per well using an Echo. Each test compound had a maximum concentration of 10 μM or 1 μM, and a three-fold serial dilution was performed over a concentration range of 10 μM to 0.2 nM. Control wells containing assay buffer without enzyme served as low controls (LCs), and wells containing vehicle (1% DMSO) reacted with the enzyme but without compound treatment served as high controls (HCs). Compounds were incubated with MALT1 enzyme and substrate at room temperature for 15 hours. Fluorescence was then measured using Envision at excitation 360 nm and emission 450 nm. Inhibition curves were fitted using XLfit and IC was calculated. 50 value, IC50 The values ​​are shown in Table 1.

[0103] Calculate IC using the following formula 50 Value (Z prime>0.5):

[0104] LC = median of low control values

[0105] Low control: reaction without MALT1 enzyme

[0106] HC = median of high control values

[0107] High control: vehicle control without compound

[0108] Inhibition % = 100 - [(sample-LC) / (HC-LC) × 100]

[0109] Curve fitting formula: fit = (A + ((BA) / (1 + ((C / x)^D))))

[0110] A: Min(Bottom), B: Max(Top), C: IC 50 (inflection point), D: slope (Hill value)

[0111] Table 1

[0112] Experimental results: IC of compound 1 of the present disclosure for inhibition of MALT1 biochemical protease 50 The value was 34 nM, which showed good inhibitory activity.

[0113] Test Example 2: Compound Solubility Determination

[0114] 1. Preparation of buffer and physiological medium

[0115] (1) Preparation of FaSSIF solution:

[0116] Preparation method of simulated fasting state intestinal fluid (FaSSIF):

[0117] Step 1: Dissolve 0.420g of sodium hydroxide, 4.470g of disodium hydrogen phosphate, and 6.186g of sodium chloride in 900mL of purified water. Adjust the pH to 6.50 with 1N sodium hydroxide solution or 1N HCl. Dilute to 1000mL with purified water at room temperature and shake well.

[0118] Step 2: Add 2.240 g of FaSSIF / FeSSIF / FaSSGF powder (Brand: Biorelevant) to 500 mL of the above buffer solution and stir until the powder is completely dissolved. Add buffer solution to the volume to 1000 mL at room temperature.

[0119] (2) Preparation of buffer solutions of different pH values:

[0120] pH 1.0 hydrochloric acid solution: Take 0.9 mL of concentrated hydrochloric acid, add it to 100 mL of ultrapure water, and shake well to obtain a pH 1.0 hydrochloric acid solution.

[0121] pH 3.0-8.0 phosphate buffer: Weigh 14.2732 g of disodium hydrogen phosphate and add water to 500 mL to obtain a 200 mM disodium hydrogen phosphate aqueous solution, shake well to dissolve as Solution A. Weigh 4.2145 g of citric acid monohydrate and add water to 200 mL, shake well to obtain Solution B. Take different volumes of Solution A and Solution B and mix them to obtain phosphate buffers of pH 3.0, 4.0, 5.0, 6.0, 6.8, 7.4, and 8.0.

[0122] 2 Determination of equilibrium solubility

[0123] The equilibrium solubility of Compound 1 and Compound 2 was determined using the standard shake flask method.

[0124] 3 Experimental results

[0125] Conclusion: In different pH and simulated biological media, the water solubility of compound 2 is significantly improved compared with compound 1.

[0126] Test Example 3: Pharmacokinetic Evaluation of Compounds 1 and 2

[0127] SD rats were used as test animals, and the plasma concentration of Compound 1 at different time points after oral administration of Compound 1 and Compound 2 was determined by LC / MS / MS. The pharmacokinetic behavior of the disclosed compounds in rats was studied, and their pharmacokinetic characteristics were evaluated.

[0128] 1 Experimental plan

[0129] 1.1 Preparation of test samples

[0130] Accurately weigh appropriate amounts of compound 1 and compound 2, add appropriate volume of 0.5% MC (methylcellulose), vortex and ultrasonicate to fully mix, and prepare the required dosage formulation for oral administration by gavage.

[0131] 1.2 Experimental animals

[0132] SD rats, SPF grade, were transferred from the experimental institution animal reserve (999M-017) of the Experimental Animal Management Department of Shanghai Institute of Family Planning Science, 4 females.

[0133] 1.3 Administration

[0134] Weigh the animals before administration and calculate the dosage based on body weight. Administer the drug by oral gavage.

[0135] 1.4 Blood collection time

[0136] 0.25h, 0.5h, 1h, 2h, 4h, 6h, 8h, 12h, 24h after administration.

[0137] 1.5 Sample collection and disposal

[0138] Blood was collected from the jugular vein, with approximately 200 μL of sample collected for each test. The sample was anticoagulated with EDTA-K2 and placed on ice. Plasma was separated by centrifugation within 1 hour (centrifugation conditions: 6800 g, 6 minutes, 2-8°C). The collected plasma samples were stored at -70°C until analysis. After analysis, the remaining plasma samples were stored at -70°C for one month.

[0139] 2. Bioanalysis and data processing

[0140] The concentration of compound 1 in plasma samples was detected, and metabolites with known ion-pair mass-to-charge ratio information were semi-quantitatively analyzed. The accuracy of quality control samples was evaluated while analyzing the samples, and it was required that the accuracy of more than 66% of the quality control samples be between 80-120%.

[0141] When plotting plasma drug concentration-time curves, BLQ is recorded as 0. When calculating pharmacokinetic parameters, the concentration before administration is calculated as 0; the BLQ before Cmax (including "No peak") is calculated as 0; and the BLQ after Cmax (including "No peak") is not included in the calculation.

[0142] WinNonlin was used to calculate pharmacokinetic parameters such as AUC(0-t), T1 / 2, Cmax, Tmax and MRT based on the blood drug concentration data at different time points.

[0143] 3 Experimental results

[0144] The pharmacokinetic parameters of the disclosed compounds are as follows: Conclusion: The pharmacokinetic properties of compound 2 (phosphate prodrug of compound 1) of the present disclosure are compared with compound 1, AUC last / D It increased by 3.8 times (10360.06 vs 2147.60), with obvious pharmacokinetic advantages.

Claims

1. A prodrug of a compound of formula I or a pharmaceutically acceptable salt thereof, 2. The prodrug or pharmaceutically acceptable salt thereof according to claim 1, wherein the prodrug is a compound represented by formula II-a, n is selected from integers of 1-5, preferably n is selected from integers of 1-3.

3. The prodrug or pharmaceutically acceptable salt thereof according to claim 1, wherein the prodrug is a compound represented by formula III, 4. The prodrug or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 3, wherein the pharmaceutically acceptable salt is a sodium salt.

5. The prodrug according to any one of claims 1 to 3 or a pharmaceutically acceptable salt thereof, wherein the prodrug is 6. The isotope-substituted prodrug or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 5, wherein the isotope-substituted prodrug is preferably a deuterium atom-substituted prodrug.

7. A pharmaceutical composition comprising the prodrug according to any one of claims 1 to 5 or a pharmaceutically acceptable salt thereof, or the isotope substitution according to claim 6, and a pharmaceutically acceptable excipient.

8. Use of the prodrug according to any one of claims 1 to 5 or a pharmaceutically acceptable salt thereof, the isotope-substituted compound according to claim 6, or the pharmaceutical composition according to claim 7 in the preparation of a medicament for preventing and / or treating a disorder associated with MALT1.

9. Use of the prodrug according to any one of claims 1 to 5 or a pharmaceutically acceptable salt thereof, the isotope-substituted product according to claim 6, or the pharmaceutical composition according to claim 7 in the preparation of a medicament for preventing and / or treating autoimmune diseases, inflammatory diseases, cancers, and tumors.

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