Alpha-hydroxy carboxylic acid derivative with adamantyl as well as preparation method and application of alpha-hydroxy carboxylic acid derivative

Abstract

The invention is applicable to the field of medicinal chemistry, and provides an alpha-hydroxy carboxylic acid derivative with adamantyl, a preparation method and application thereof, and a stereoisomer or pharmaceutically acceptable salt thereof, the derivative has the following structural general formula I: # imgabs0 #, in the general formula, when X = O, n = 0 or 1, and A is a lipophilic group or a hydrophilic group; when X is equal to N in the general formula, n is equal to 0-3, and A is one of alkyl, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, aromatic heterocyclic group, heteroaromatic heterocyclic group, pyrrolidine, piperazine, piperidine, morpholine group, other cyclic amines and substituted derivatives of the groups. The compound has a structural formula as shown in a formula I, has good inhibitory activity on influenza viruses, can be developed into a new generation of M2 inhibitors, and lays a good foundation for development of anti-influenza drugs.

Description

Technical Field

[0001] The invention belongs to the field of pharmaceutical chemistry, and in particular relates to an alpha-hydroxycarboxylic acid derivative having an adamantyl group, and a preparation method and application thereof. Background Art

[0002] Influenza virus is the culprit of seasonal influenza (flu) and occasional influenza pandemics every year, impacting the public health safety system. Currently, vaccination is the most effective way to prevent influenza, but it can only prevent it, while treatment requires drugs.

[0003] At present, the treatment drugs recommended by the China Center for Disease Control and Prevention include neuraminidase (NA) inhibitors, hemagglutinin inhibitor (HA) arbidol, and RNA polymerase inhibitors favipiravir and mabaloxavir. M2 inhibitors (amantadine and rimantadine) are no longer recommended for clinical use due to drug resistance issues. However, the parasitic characteristics of influenza viruses and the variability of their antigens make it difficult for these chemotherapeutic drugs to continue to work, resulting in varying degrees of drug resistance. Some seasonal influenza A viruses (H1N1) are resistant to oseltamivir and peramivir, and resistant strains of mabaloxavir have also been isolated clinically. Combination therapy of anti-influenza virus drugs with different mechanisms of action can enhance the therapeutic effect or reduce the emergence of drug-resistant strains. Since the development trend of influenza and the mutation of the virus are still unpredictable, the development of anti-influenza drugs is a hot direction in the field of drug research and development, and it is also the first line of defense for humans to fight against influenza in order to cope with possible influenza pandemics.

[0004] The main cause of resistance to adamantane M2 inhibitors is mutations in the transmembrane region of the M2 protein, especially the S31N variant, whose specific mechanism of action is that the residue serine (Ser) on the M2 ion channel mutates to glutamine (Asn), which reduces the size of the drug binding site, prevents the drugs adamantane and rimantadine from binding in the pore and blocks proton conduction. At present, the literature has reported adamantane inhibitor for M2-S31N resistance, which was discovered by the DeGrado and Wang research groups in the United States, and summarized the pharmacophore model of M2-S31N inhibitors, and reported a series of adamantane analogs. The common characteristics of this type of inhibitors are: hydrophobic skeleton + methylene bridge + hydrophobic five (six) membered aromatic ring.

[0005] From the perspective of drug design, adamantane is considered a "liphophilic bullet" that can increase the hydrophobicity and structural stability of the drug. It also has no rotatable bonds and only loses a very small amount of conformational entropy when binding to the target protein. In addition, adamantane derivatives are a hot topic in the field of antiviral drug research and development. For example, adamantane derivatives can treat influenza, herpes simplex virus, hepatitis C virus and AIDS.

[0006] Therefore, based on the structural characteristics of M2-S31N inhibitors and adamantane ethylamine reported in the literature, the designed novel adamantane derivatives are expected to be developed into a new generation of M2 inhibitor anti-influenza drugs. Summary of the invention

[0007] The purpose of the embodiments of the present invention is to provide an α-hydroxycarboxylic acid derivative having an adamantyl group and a preparation method and application thereof, which has good inhibitory activity against influenza virus.

[0008] The present invention is achieved by an α-hydroxycarboxylic acid derivative having an adamantyl group, and its stereoisomers or pharmaceutically acceptable salts have the following structural formula I:

[0009]

[0010] Wherein, when X=O in the general formula, n=0 or 1, A is a lipophilic group or a hydrophilic group;

[0011] In the general formula, when X=N, n=0-3, A is an alkyl group, a cycloalkyl group, a heterocycloalkyl group, an aryl group, a heteroaryl group, a cyclic aromatic group, a heterocyclic aromatic group, a tetrahydropyrrole group, a piperazine group, a piperidine group, a morpholinyl group and other cyclic amines, and one of the substituted derivatives of the above groups.

[0012] A further technical solution is selected from any one of the following structural formulas:

[0013]

[0014] A method for preparing an α-hydroxycarboxylic acid derivative having an adamantyl group, wherein the synthetic route is:

[0015]

[0016] The following steps are involved:

[0017] Step 1: using 2-adamantyl-α-keto acid 1 as a raw material and cis-N-toluenesulfonylaminoindanol 2 under the action of a condensing agent to generate an ester intermediate 3;

[0018] Step 2: Compound 3 obtained in step 1 undergoes a nucleophilic addition reaction with 2-methylthiophene lithium prepared in situ at low temperature to obtain compound 5;

[0019] Step 3, the compound 5 obtained in the above step 2 is hydrolyzed under the conditions of LiOH / CH3OH / THF to obtain a carboxylic acid derivative 6;

[0020] Step 4: Take the carboxylic acid compound 6 obtained in the above step 3 and condense it with differently substituted primary amines (R-NH2) via a condensing agent to obtain target compounds Ⅰ-1~Ⅰ-4, Ⅰ-7~Ⅰ-17; or react compound 6 with a bromide under NaHCO3 to generate ester compounds Ⅰ-5 and Ⅰ-6.

[0021] In a further technical solution, in step 1, the molar ratio of α-keto acid 1 to the catalyst is 1:05-1.5, the selected catalyst is EDCI·HCl / DMAP or TBTU, and the solvent is dichloromethane or THF.

[0022] In a further technical solution, the reaction temperature in step 2 is -78°C and the reaction time is 3-5h.

[0023] According to a further technical solution, the solvent ratio selected in step three is tetrahydrofuran: methanol: water in a volume ratio of 2:3:1, and the temperature is 60-80°C.

[0024] In a further technical solution, the catalyst used to form the amide in step 4 is CDI or EDCI·HCl / HOBt, the solvent is dichloromethane, and the temperature is room temperature 25°C.

[0025] An alpha-hydroxycarboxylic acid derivative having an adamantyl group, and use of its pharmaceutically acceptable salt, crystalline hydrate and solvate in the preparation of anti-influenza virus drugs

[0026] In a further technical solution, the influenza virus is influenza A H3N2 (A / Hong Kong / 8 / 68 strain, amantadine-sensitive strain) or H1N1 (A / WSN / 1933, amantadine-resistant strain).

[0027] Compared with the prior art, the present invention has the following beneficial effects:

[0028] The present invention provides an α-hydroxycarboxylic acid derivative with an adamantyl group, a preparation method and application thereof, with a structural formula as Formula I, which has good inhibitory activity against influenza virus and can be developed into a new generation of M2 inhibitors, laying a good foundation for the development of anti-influenza drugs; the present invention also provides a preparation method for the above-mentioned compound, which is easy to prepare a series of compounds; through in vitro anti-influenza virus experiments, it is shown that the above-mentioned compound improved by the present invention has certain inhibitory activity against both wild-type (H3N2) and variant (H1N1) influenza viruses, and has less toxic and side effects, and the activity of individual compounds is equivalent to that of the positive drug oseltamivir, showing that the compound and its pharmaceutically acceptable salts, crystalline hydrates and solvates can be used in the preparation of anti-influenza virus drugs. The present invention also provides a pharmaceutical composition comprising the above-mentioned compound, which can be prepared into dosage forms such as tablets, capsules and suspensions as needed, and has important development value and economic benefits for the prevention and treatment of influenza. DETAILED DESCRIPTION

[0029] In order to make the purpose, technical solution and advantages of the present invention more clearly understood, the present invention is further described in detail below in conjunction with the embodiments. It should be understood that the specific embodiments described herein are only used to explain the present invention and are not used to limit the present invention.

[0030] The specific implementation of the present invention is described in detail below in conjunction with specific embodiments.

[0031] The term "pharmaceutically acceptable salt" means that the α-hydroxycarboxylic acid derivatives I-7 to I-11 having an adamantyl group are basic and can react with inorganic or organic acids to form conventional non-toxic salts, for example, hydrochloride, hydrobromide, sulfate, nitrate, phosphate, methanesulfonate, benzenesulfonate, p-toluenesulfonate, oxalate, citrate, mandelate, nicotinate, tartrate, palmitate, malate, succinate, maleate.

[0032] The term "stereoisomers" refers to compounds that have identical chemical formula, but differ in the arrangement of the atoms or groups in space.

[0033] A pharmaceutical composition containing an α-hydroxycarboxylic acid derivative having an adamantyl group and at least one pharmaceutically acceptable pharmaceutical excipient, comprising an α-hydroxycarboxylic acid derivative having an adamantyl group as shown in Formula I and pharmaceutically acceptable salts, crystalline hydrates and solvates, and stereoisomers thereof. The "pharmaceutical excipient" refers to various excipients conventionally used in drugs, such as excipients, diluents, controlled release agents, stabilizers, disintegrants, wetting agents, surfactants, etc.

[0034] Basic step, synthesis of 2-(adamantan-1-yl)-2-oxoacetic acid (1)

[0035]

[0036] To a round-bottom flask (250 mL), add 1-adamantyl ketone (3.0 g, 17 mmol, 1 equiv) and water (50 mL). At 0°C, add pyridine (4.1 mL, 51 mmol, 3 equiv), potassium hydroxide (KOH, 1.12 g, 20 mmol, 1.18 equiv), and add solid potassium permanganate (KMnO4, 5.37 g, 34 mmol, 2 equiv) in batches. After the addition, stir with electromagnetic at 60°C, install a reflux device, and place a balloon to absorb the gas. Monitor the reaction by thin layer chromatography (TLC). After 3 h, the reaction is complete. Filter while hot, and wash the filter flask with hot water (10 mL×3). The filtrate was combined, concentrated hydrochloric acid was added to adjust the pH to 1, and the mixture was extracted with ethyl acetate (50 mL × 3). The organic phase was washed with saturated brine (80 mL × 2), dried over anhydrous Na2SO4, filtered using a small funnel plugged with cotton wool, and the solvent was removed by distillation under reduced pressure to obtain adamantane-1-ylglyoxylic acid intermediate 1 (3.22 g, yield 91.0%) as a white solid.

[0037] Synthesis of (1S,2R)-1-((4-methylphenyl)sulfonamido)-2,3-dihydro-1H-inden-2-yl 2-(adamantan-1-yl)-2-oxoacetate (3)

[0038]

[0039] In a round-bottom flask (250 mL), α-keto acid 1 (2.08 g, 10 mmol, 1 equiv) obtained in the previous step and dichloromethane (100 mL) were added, stirred at 0°C, EDC·HCl (2.875 g, 15 mmol, 1.5 equiv), 4-dimethylaminopyridine (DMAP, 2.44 g, 20 mmol, 2 equiv) were added, and finally cis-N-toluenesulfonylaminoindanol 7 (3.03 g, 10 mmol, 1 equiv) was added in batches, and stirred at room temperature. The reaction was monitored by thin layer chromatography, and the reaction was complete after 12 h. The mixture was extracted with ethyl acetate (50 mL × 3), and the organic phase was washed once with dilute hydrochloric acid to remove the organic base, and then washed with saturated brine (80 mL × 2), dried over anhydrous Na2SO4, filtered with a small funnel plugged with cotton wool, and concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (petroleum ether / ethyl acetate, 15:1-1:1) to give intermediate 3 (3.0 g, yield 60.8%). 1HNMR(400MHz,Chloroform-d)δ8.33(d,J=8.8Hz,1H),7.79(d,J=8.2Hz,2H),7.40(d, J=8.2Hz,2H),7.26-7.18(m,2H),7.13(t,J=7.5Hz,1H),6.82(d,J=7.5Hz,1H),5.42(t d,J=5.0,1.5Hz,1H),5.05(dd,J=8.8,5.0Hz,1H),3.25(dd,J=17.2,5.0Hz,1H),2.87( d,J=17.2,1H),2.40(s,3H),1.98-1.93(m,3H),1.80-1.78(m,3H),1.75-1.57(m,9H). 13 C NMR (100MHz, CDCl3) δ200.5,162.6,144.0,139.3,138.1,137.8,130.0,128.9 ,127.8,127.2,125.1,124.3,76.6,59.9,45.1,37.5,37.2,36.3,27.6,21.7.

[0040] Synthesis of (1S,2R)-1-((4-methylphenyl)sulfonamido)-2,3-dihydro-1H-inden-2-yl(S)-2-(adamantan-1-yl)-2-hydroxy-2-(5-methylthiophen-2-yl)acetic acid ethyl ester (5)

[0041]

[0042] To a round-bottom flask (250 mL), add ester compound 3 (3.46 g, 7 mmol, 1 equiv), add anhydrous tetrahydrofuran (20 mL) in a glove box under anhydrous and oxygen-free environment to dissolve it. To another round-bottom flask (25 mL), add 2-methylthiophene (1.38 g, 14 mmol, 2 equiv), add anhydrous tetrahydrofuran (100 mL) in a glove box under anhydrous and oxygen-free environment, place at -78 ° C, cool for a few minutes, then slowly add n-butyl lithium (4.2 mL, 2.5 mol / L), and continue to react at -78 ° C. After reacting for 1 hour. Aspirate the raw material 7 with a syringe and slowly add it to the round-bottom flask. Monitor the reaction by thin layer chromatography. After 3 hours, the reaction is complete. The reaction was quenched with an appropriate amount of ammonium chloride solution, extracted with ethyl acetate (80 mL×3), and the organic phase was washed with saturated brine (150 mL×2), dried over anhydrous Na2SO4, filtered with a small funnel plugged with cotton wool, and the solvent was removed by rotary evaporation under reduced pressure to obtain a crude product, which was purified by silica gel column chromatography (eluent: petroleum ether / ethyl acetate, 15:1-6:1) to obtain intermediate 5 (3.60 g, yield 86.9%) as white crystals.1 HNMR(400MHz,DMSO-d6)δ8.30(d,J=10.2Hz,1H),7.56(d,J=8.2Hz,2H),7.27-7.23(m,2H),7.21- 7.18(m,2H),7.01(d,J=8.2Hz,2H),6.80(d,J=3.5Hz,1H),6.74(dd,J=3.5,1.2Hz,1H),5.91(s,1H ),4.97(dd,J=10.1,4.8Hz,1H),4.77(t,J=4.6Hz,1H),3.13(dd,J=17.2,4.5Hz,1H),2.74(d,J=17 .2Hz,1H),2.45(s,3H),2.28(s,3H),1.83(s,3H),1.54(d,J=12.6Hz,6H),1.41(d,J=12.6Hz,6H). 13 C NMR (100MHz, CDCl3) δ173.2,144.1,139.4,139.4,139.1,138.7,137.6,130.1,128.9,127.7,127.2,126.0 ,125.2,124.5,124.0,83.0,79.1,59.8,40.6,37.1,36.8,36.6,28.6,21.7,15.1.LC-MS(ESI)m / z:[M+Na] + Calcd for C 33 H 37 NO5S2,614.21,Found 614.0.

[0043] Synthesis of 2-(adamantan-1-yl)-2-hydroxy-2-(5-methylthiophen-2-yl)acetic acid 9

[0044]

[0045] In a round-bottom flask (250 mL), add the ester compound 5 (3.6 g, 6.09 mmol, 1 equiv) obtained in the previous step, tetrahydrofuran (40 mL), methanol (60 mL), then add LiOH·H2O (1.28 g, 30.45 mmol, 5 equiv), water (20 mL), stir at 60 ° C with electromagnetic stirring, and install a reflux device. Monitor the reaction by thin layer chromatography. After 24 hours, the reaction is complete. Remove tetrahydrofuran by vacuum distillation, add water, dichloromethane and 10% sodium hydroxide solution for extraction, and retain the aqueous phase. The aqueous phase was adjusted to pH 1 by adding dilute hydrochloric acid, and the mixture was extracted with ethyl acetate (10 mL × 3). The organic phase was washed with saturated brine (20 mL × 2), dried over anhydrous Na2SO4, filtered through a small funnel plugged with cotton wool, and concentrated under reduced pressure to give 2-(adamantan-1-yl)-2-hydroxy-2-(5-methylthiophen-2-yl)acetic acid 6 (65 mg, yield 96.6%). 1 H NMR(400mHz,DMSO-d6)δ6.86(d,J=3.5Hz,1H),6.63(dd,J=3.5,1.2Hz,1H),2.36(s,3H), 1.91(t,J=3.3Hz,3H),1.76(d,J=12.0Hz,3H),1.63–1.52(m,6H),1.48(d,J=12.0Hz,3H). 13 CNMR(100mHz,DMSO)δ174.3,141.3,137.6,125.4,124.4,81.4,36.5,36.0,28.0,14.7,14.6.

[0046] Example 1, Synthesis of 2-(adamantan-1-yl)-2-hydroxy-2-(5-methylthiophen-2-yl)-N-(2-morpholinoethyl)acetamide (I-9)

[0047]

[0048] In a round-bottom flask (25 mL), add the raw material 6 (122.4 mg, 0.4 mmol, 1 equiv), dissolve it in an appropriate amount of dichloromethane, and add N, N'-carbonyldiimidazole (CDI, 104.7 mg, 0.8 mmol, 2 equiv) in portions. Stir at room temperature with electromagnetic stirring for 2 h. Add 4-(2-aminoethyl)morpholine 10 (104.7 mg, 0.8 mmol, 2 equiv) and continue to react at room temperature. Monitor the reaction by thin layer chromatography. After 5 h, the reaction is complete. Extract with dichloromethane (15 mL × 3), wash the organic phase with saturated brine (20 mL × 2), dry with anhydrous Na2SO4, filter with a small funnel plugged with cotton wool, and evaporate under reduced pressure to obtain a yellow oil. Purify the crude product by silica gel column chromatography (petroleum ether / ethyl acetate, 5:1-0:1) to obtain the target product Ⅰ-9 (91 mg, yield 54.5%) as a white powder.

[0049] LC-MS(ESI)m / z:[M+H] + Calcd for C 23 H 34 N2O3S,592.21,Found,592.1; IR(KBr)ν(cm -1 ): 3430, 3418, 3402, 3054, 2907, 2850, 2801, 1651, 1526, 1473, 1452, 1363, 1344, 1117; 1HNMR (400m Hz, CDCl3) δ7.24(t,J=5.2Hz,1H),6.95(d,J=3.5Hz,1H),6.63(dd,J=3.5,1.2Hz,1H),3.67(t,J=4. 6Hz,4H),3.45-3.37(m,1H),3.30(dt,J=19.6,5.8Hz,2H),2.48(td,J=5.6,1.2Hz,2H),2.44-2.42 (m,7H),1.98(p,J=3.0Hz,3H),1.85(dq,J=12.2,2.3Hz,3H),1.69-1.63(m,6H),1.60-1.55(m,3H); 13 C NMR (100mHz, CDCl3) δ172.3,140.8,139.7,125.4,124.3,82.9,67.1,57.0,53.4,40.5,36.9,36.5,35.8,28.7,15.1.

[0050] Example 2, Synthesis of 2-(adamantan-1-yl)-2-hydroxy-2-(5-methylthiophen-2-yl)-N-((bicyclo[2.2.1]heptane-2-yl)methyl)acetamide (I-4)

[0051] Carboxylic acid 6 (94.8 mg, 0.31 mmol), 5-norbornene-2-methylamine (40.1 mg, 0.3255 mmol, 1.05 equiv.) and 3 mL of dichloromethane were added to a 25 mL round-bottom flask, stirred at room temperature to dissolve, and then EDCI·HCl (65.4 mg, 0.341 mmol, 1.1 equiv) and HOBt (46.1 mg, 0.341 mmol, 1.1 equiv) were added in sequence, and reacted at room temperature for 6 hours. An appropriate amount of water was added to quench the reaction, and dichloromethane was extracted 3 times (15 mL). The organic phase was washed twice with dilute hydrochloric acid and twice with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The product 185 mg was obtained by silica gel column chromatography as a white solid with a yield of 59.7%.

[0052] 1 H NMR(400mHz,DMSO-d6)δ7.60(q,J=5.2Hz,1H),6.87(d,J=3.5Hz,1H),6.61( d,J=3.5Hz,1H),6.16–6.14(m,1H),6.05–5.98(m,1H),5.83(s,1H),2.86–2. 62(m,3H),2.36(s,3H),2.29–2.16(m,1H),1.89(s,3H),1.83-1.71(m,4H),1 .61-1.42(m,10H),1.35-1.26(m,1H),1.23-1.17(m,1H),0.53-0.48(m,1H). 13 C NMR (100mHz, CDC l3 )δ171.9,140.5,139.8,137.8,132.2,125.5,124.3,82.9,49.6,44.5,43.5,42.5,40.5,39.0,38.8,36.9,36.5,28.7,15.1.

[0053] Example 3 Synthesis of 2-(adamantan-1-yl)-2-hydroxy-2-(5-methylthiophen-2-yl)-N-(propargyl-1-yl)acetamide (I-1)

[0054] The 4-(2-aminoethyl)morpholine in Example 1 was replaced by propargylamine, and the other operation steps were the same as in Example 1 to synthesize the target compound as a white powder with a yield of 77.6%. 1H NMR (400mHz, CDCl3) δ7.43–7.22(m,1H),7.17(d,J=7.3Hz,1H),6.99(d,J=3.5Hz,1H),6.67(d,J=3.5Hz,1H),4.18–4.11(m,1H) ,4.07–4.00(m,1H),2.79(s,1H),2.47(s,3H),2.25(t,J=2.6Hz,1H),2.07-2.00(m,3H),1.96-1.88(m,3H),1.73-1.61(m,9H). 13 CNMR (100mHz, CDCl3) δ172.1,140.1,139.6,125.8,124.4,83.1,79.5,71.8,40.7,36.8,36.4,29.1,28.7,15.2.

[0055] Example 4. Synthesis of 2-(adamantan-1-yl)-2-hydroxy-2-(5-methylthiophen-2-yl)-N-((tetrahydrofuran-3-yl)methyl)acetamide (I-2)

[0056] The 5-norbornene-2-methylamine in Example 2 was replaced with 3-aminomethyl-tetrahydrofuran, and the other operation steps were the same as those in Example 2. The target compound was synthesized as a white solid with a yield of 78.8%.

[0057] 1 H NMR (400mHz, CDCl3) δ7.09 (s, 1H), 6.94 (d, J = 3.5Hz, 1H), 6.63 (d, J = 3.5Hz, 1H), 3.88–3.82 (m, 1H), 3.82–3.67 (m, 2H), 3.50 (dd, J = 8.2, 5. 5Hz,1H),3.39–3.16(m,2H),2.90–2.88(m,1H),2.54–2.45(m,1H),2.43(s,3H),2.03-1.98(m,4H),1.92-1.82(m,3H),1.67-1.58(m,10H). 13 C NMR (100mHz, CDCl3) δ172.5,139.9,125.6,124.3,83.0,71.6,71.5,67.9,67.9,42.3,40.5,39.2,39.2,36.8,36.5,30.2,28.7,15.2.

[0058] Example 5. Synthesis of 2-(adamantan-1-yl)-2-hydroxy-2-(5-methylthiophen-2-yl)-N-((1R,2R,3R,5S)-2,6,6-trimethylbicyclo[3.1.1]heptane-3-yl)acetamide (I-3)

[0059] The 4-(2-aminoethyl)morpholine in Example 1 was replaced with (1R,2R,3R,5S)-3-pinaneamine, and the other operation steps were the same as in Example 1. The target compound was synthesized as a white powder with a yield of 26.3%.

[0060] 1 H NMR(300mHz, CDCl3) δ6.95(d,J=3.5Hz,1H),6.77(d,J=8.6Hz,1H),6.64(d,J=3.6Hz,1H),4.3 4-4.23(m,1H),2.89(s,1H),2.67-2.57(m,1H),2.44(s,3H),2.41-2.37(m,1H),2.00-1.95(m ,3H),1.89(d,J=12.5Hz,3H),1.84-1.75(m,2H),1.65(d,J=16.0Hz,6H),1.51(dd,J=6.0,2.0 Hz,1H),1.27(d,J=9.2Hz,1H),1.22(s,3H),1.07(s,3H),1.05(s,3H),0.85(d,J=9.8Hz,1H). 13 C NMR (100mHz, CDCl3) δ171.3,140.3,140.1,125.6,124.2,82.9,48.0,47.6,46. 5,41.7,40.5,38.6,37.1,36.9,36.6,35.4,29.8,28.7,28.1,23.4,21.1,15.2.

[0061] Example 6. Synthesis of ((prop-1-en-2-yl)cyclohex-1-en-1-yl)methyl 2-(adamantan-1-yl)-2-hydroxy-2-(5-methylthiophen-2-yl)acetate (I-5);

[0062]

[0063] In a 100mL round-bottom flask, add perillyl alcohol (456.7mg, 3mmol), triphenylphosphine (865.6mg, 3.3mmol) and 20mL of dichloromethane, cool the reaction solution to 0°C, and add NBS (587.4mg, 3.3mmol) in batches under magnetic stirring. After the addition is completed, remove the ice-water bath and stir at room temperature for 1h. After the reaction is completed, remove the solvent under reduced pressure, then add n-hexane several times, solid precipitates, filter, remove the solvent from the filtrate under reduced pressure, and obtain 464mg of its bromide as a colorless liquid by silica gel column chromatography (eluent is n-hexane), with a yield of 72.0%.

[0064]

[0065] Add raw material 6 (54 mg, 0.176 mmol) and the bromide obtained in the previous step (75 mg, 0.352 mmol) and 1 mL of DMF to a 25 mL round-bottom flask, stir at room temperature, add NaHCO3 (29.6 mg, 0.352 mmol) at once, stir at room temperature for 6 hours in a dark place, TLC shows that the reaction is complete. Transfer to a separatory funnel and extract three times with 10 mL of ethyl acetate, wash twice with water, wash twice with saturated brine, dry with anhydrous sodium sulfate, filter, evaporate the solvent, and obtain the residue by silica gel column chromatography to obtain the target compound (I-5) as a colorless viscous liquid 47 mg, with a yield of 60.6%.

[0066] 1 H NMR(400mHz, CDCl3) δ6.90(d,J=3.5Hz,1H),6.63(d,J=3.5Hz,1H),5.87(s,1H),4.79-4.67(m,3H),4.63-4.55(m,1H),4.05 (s,1H),2.44(s,3H),2.20-2.12(m,4H),2.02-1.95(m,4H),1.93-1.78(m,5H),1.75(s,3H),1.68-1.61(m,6H),1.55(s,2H). 13 C NMR (75mHz, CDCl3) δ174.0,149.3,140.3,138.9,131.8,128.1,125.8,124.7, 109.0,82.4,70.9,40.6,36.9,36.3,30.5,28.6,27.6,27.2,26.9,20.9,15.2.

[0067] Example 7, Synthesis of (6,6-dimethylbicyclo[3.1.1]hept-2-en-2-yl-2-adamantan-1-yl)methyl 2-(adamantan-1-yl)-2-hydroxy-2-(5-methylthiophen-2-yl)acetate (I-6);

[0068] The perillyl alcohol in Example 6 was replaced by myrtenol, and the other operation steps were the same as those in Example 6. The target compound was synthesized as a colorless viscous liquid with a yield of 89.7%.

[0069] 1 H NMR(400mHz, CDCl3) δ6.87(d,J=3.5Hz,1H),6.62(dd,J=3.5,1.2Hz,1H),5.73–5.67(m,1H),4.72 (dd,J=11.8,1.2Hz,1H),4.51(dd,J=11.8,1.2Hz,1H),4.12(s,1H),2.43(s,3H),2.44-2.42(m,1H ),2.36-2.28(m,2H),2.25(td,J=5.6,1.6Hz,1H),2.13(ddq,J=4.6,3.1,1.6Hz,1H),1.98(t,J=3 .2Hz,3H),1.85-1.77(m,3H),1.68-1.55(m,9H),1.29(s,3H),1.21(d,J=8.7Hz,1H),0.85(s,3H). 13 C NMR (100mHz, CDCl3) δ174.2,142.2,140.3,138.9,125.8,124.7,124.1,82 .4,69.9,43.9,40.6,38.2,36.9,36.4,31.7,31.4,28.7,26.1,21.1,15.2.

[0070] Example 8, Synthesis of 2-(adamantan-1-yl)-2-hydroxy-2-(5-methylthiophen-2-yl)-N-(2-(4-methylpiperazin-1-yl)ethyl)acetamide (I-7);

[0071] The 5-norbornene-2-methylamine in Example 2 was replaced by 4-methyl-1-piperazineethylamine, and the other operation steps were the same as those in Example 2. The target compound was synthesized as a white solid with a yield of 37.8%.

[0072] 1H NMR (400mHz, DMSO-d6) δ7.59(t,J=5.6Hz,1H),6.86(d,J=3.5Hz,1H),6.61(dd,J=3.5,1.2Hz,1H),5.85(s,1H),3.31–3.26(m,1H),3 .09–3.02(m,1H),2.46–2.19(m,12H),2.13(s,3H),1.89(s,3H),1.75(d,J=12.0Hz,3H),1.61-1.51(m,6H),1.46(d,J=12.0Hz,3H). 13 C NMR (100mHz, CDCl3) δ172.4,140.9,139.7,125.4,124.3,82.8,56.5,55.1,52.8,46.1,40.4,36.9,36.5,35.9,28.8,15.2.

[0073] Example 9, Synthesis of 2-(adamantan-1-yl)-2-hydroxy-2-(5-methylthiophen-2-yl)-N-(1-(4-methylpiperidin-4-yl)methyl)acetamide (I-8);

[0074] The 5-norbornene-2-methylamine in Example 2 was replaced with (1-methyl-4-piperidinyl)methylamine, and the other operation steps were the same as those in Example 2. The target compound was synthesized as a white solid with a yield of 40.0%.

[0075] 1 H NMR (400mHz, DMSO-d6) δ7.62(t,J=5.8Hz,1H),6.86(d,J=3.5Hz,1H),6.61(d,J=3.5Hz,1H),5.85(s,1H),2.98(d,J=6.2Hz,2H),2.72–2.68(m,2H) ,2.36(s,3H),2.10(s,3H),1.91–1.86(m,3H),1.79-1.75(m,4H),1.74-1 .70(m,1H),1.56(d,J=12.0Hz,6H),1.51-1.43(m,6H),1.18-1.06(m,2H). 13 C NMR (100mHz, CDCl3) δ172.4,140.2,139.9,125.6,124.2,83.0,55.5,46.4,44.8,40.4,36.8,36.5,35.4,30.1,30.1,28.7,15.1.

[0076] Example 10. Synthesis of 2-(adamantan-1-yl)-2-hydroxy-2-(5-methylthiophen-2-yl)-N-((4-methylmorpholin-2-yl)methyl)acetamide (I-10)

[0077] The 4-(2-aminoethyl)morpholine in Example 1 was replaced with 4-methylmorpholine-2-methylamine, and the other operation steps were the same as in Example 1. The target compound was synthesized as a white solid with a yield of 22.8%.

[0078] 1 H NMR (400mHz, CDCl3) δ7.21 (s, 1H), 6.94 (dd, J = 3.7, 1.7Hz, 1H), 6.62 (s, 1H), 3.88– 3.82(m,1H),3.71–3.60(m,2H),3.58–3.47(m,1H),3.23–3.11(m,1H),2.74(t,J=11 .2Hz,1H),2.68(d,J=11.5Hz,1H),2.42(s,3H),2.29(s,3H),2.12(tt,J=11.6,3.3 Hz,1H),2.00-1.95(m,3H),1.92-1.85(m,4H),1.62(t,J=12.2Hz,9H),1.25(s,1H). 13 C NMR (100mHz, CDCl3) δ172.6,140.2,139.9,125.6,124.3,83.1,83.0,77.4,74.5,66.4 ,57.7,57.6,54.6,46.2,46.1,41.7,41.7,40.5,40.5,36.9,36.509,36.5,28.7,15.2.

[0079] Example 11, Synthesis of 2-(adamantan-1-yl)-2-hydroxy-2-(5-methylthiophen-2-yl)-N-(3-(pyrrolidin-1-yl)propyl)acetamide (I-11)

[0080] The 4-(2-aminoethyl)morpholine in Example 1 was replaced with 1-(3-aminopropyl)pyrrolidine, and the other operation steps were the same as in Example 1. The target compound was synthesized as a white foamy solid with a yield of 58.2%. 1H NMR (400mHz, DMSO-d6) δ7.96(t,J=6.0Hz,1H),6.88(d,J=3.5Hz,1H),6.62(d,J=3.5Hz,1H),5.89(s,1H),3.19 –3.14(m,3H),3.00(t,J=6.8Hz,3H),2.36(s,3H),1.89(s,8H),1.80(s,3H),1.77(s,2H),1.61-1.42(m,10H). 13 C NMR (75mHz, CDCl3) δ174.3,141.6,140.3,126.6,124.9,83.6,78.5,78.1,77.6,54.6,41.1,38.6,37.8,29.6,27.8,24.2,16.1.

[0081] Example 12: Synthesis of 2-(adamantan-1-yl)-2-hydroxy-2-(5-methylthiophen-2-yl)-N-((1S,4S)-4-hydroxycyclohexane)acetamide (I-12)

[0082] The 5-norbornene-2-methylamine in Example 2 was replaced by cis-4-aminocyclohexanol hydrochloride, and the other operation steps were the same as those in Example 2. The target compound was synthesized as a white solid with a yield of 68.8%. 1 H NMR (400mHz, DMSO-d6) δ7.23(d,J=8.0Hz,1H),6.87(d,J=3.5Hz,1H),6.62(d,J=3.5Hz,1H),5.91(s,1H),4.4 2(d,J=3.0Hz,1H),3.70–3.57(m,2H),2.36(s,3H),1.89(s,3H),1.80(d,J=12.0Hz,3H),1.62-1.44(m,17H). 13 C NMR (100mHz, DMSO-d6) δ170.8,139.8,138.0,125.5,123.2,81.1,64.3,45.3,35.5,30.6,30.6,27.7,26.8,26.4,14.3.

[0083] Example 13. Synthesis of 2-(adamantan-1-yl)-2-hydroxy-2-(5-methylthiophen-2-yl)-N-(2-hydroxyphenethyl)acetamide (I-13)

[0084] The 4-(2-aminoethyl)morpholine in Example 1 was replaced by 2-(2-aminoethyl)phenol, and the other operation steps were the same as those in Example 1. The target compound was synthesized as a white solid with a yield of 44.2%.1 H NMR (400mHz, DMSO-d6) δ9.38 (s, 1H), 7.65 (t, J = 5.4Hz, 1H), 7.06 (dd, J = 7.4, 1.2Hz, 1H), 7.00 (td, J = 7.8, 1.2Hz, 1H), 6.86 (d, J = 3.5Hz, 1H), 6.78 (dd, J = 8.0, 1.2Hz, 1H), 6.70 (td, J = 7.8, 1.2Hz, 1H), 6.61 (dd, J = 3. 5,1.2Hz,1H),5.82(s,1H),3.46-3.37(m,1H),3.24-3.17(m,1H),2.76-2.64(m,2H),2.36(s,3H),1.91-1 .81(m,3H),1.72(d,J=12.4Hz,3H),1.55(d,J=12.1Hz,3H),1.44(t,J=12.1Hz,6H),1.17(t,J=7.2Hz,1H). 13 C NMR(100mHz,DMSO-d6)δ172.2,155.3,140.4,138.4,130.1,127.2,125.9,125.4, 123.5,118.9,114.89,81.7,59.8,39.4,36.6,35.9,29.6,28.1,20.8,14.7,14.1.

[0085] Example 14. Synthesis of 2-(adamantan-1-yl)-2-hydroxy-2-(5-methylthiophen-2-yl)-N-(3-hydroxy-4-methoxyphenethyl)acetamide (I-14)

[0086] The 5-norbornene-2-methylamine in Example 2 was replaced by 5-(2-aminoethyl)-2-methoxyphenol, and the other operation steps were the same as those in Example 2. The target compound synthesized was a white solid with a yield of 38.1%.

[0087] 1H NMR (400mHz, DMSO-d6) δ8.83 (s, 1H), 7.63 (t, J = 5.4Hz, 1H), 6.86 (d, J = 3.5Hz, 1H), 6.80 (d, J=8.2Hz,1H),6.65(d,J=2.1Hz,1H),6.61(d,J=3.5Hz,1H),6.58(dd,J=8.2,2.1Hz,1H),5. 81(s,1H),3.72(s,3H),3.44-3.37(m,1H),3.20-3.11(m,1H),2.59(t,J=7.0Hz,2H),2.36( s,3H),1.86(s,3H),1.72(d,J=12.0Hz,3H),1.55(d,J=12.0Hz,3H),1.46(d,J=10.8Hz,6H). 13 C NMR(100mHz,DMSO-d6)δ172.1,146.4,146.1,140.4,138.3,131.8,125.9,1 23.6,119.0,115.9,112.2,81.7,55.6,39.4,36.6,35.9,34.5,28.1,14.7.

[0088] Example 15. Synthesis of 2-(adamantan-1-yl)-2-hydroxy-2-(5-methylthiophen-2-yl)-N-(2-(1H-indol-3-yl)ethyl)acetamide (I-15)

[0089] The 5-norbornene-2-methylamine in Example 2 was replaced by tryptamine, and the other operation steps were the same as those in Example 2. The target compound synthesized was a white solid with a yield of 61.2%.

[0090] 1 H NMR (400mHz, DMSO-d6) δ10.81(s,1H),7.75(t,J=5.6Hz,1H),7.57(d,J=7.8Hz,1H),7.32(d ,J=8.0Hz,1H),7.16(d,J=1.2Hz,1H),7.06(t,J=7.2Hz,1H),6.97(t,J=7.2Hz,1H),6.87(d, J=3.5Hz,1H),6.62(dd,J=3.5,1.2Hz,1H),5.83(s,1H),3.52-3.44(m,1H),3.37-3.28(m,1 H),2.90-2.79(m,2H),2.36(s,3H),1.86(s,3H),1.74(d,J=12.0Hz,3H),1.56-1.41(m,9H). 13C NMR (100mHz, DMSO-d6) δ172.2,140.4,138.4,136.3,127.2,125.9,123.6,122. 7,120.9,118.4,118.2,111.6,111.3,81.7,39.5,36.6,35.9,28.1,25.2,14.7.

[0091] Example 16. Synthesis of 2-(adamantan-1-yl)-2-hydroxy-2-(5-methylthiophen-2-yl)-N-(2-(5-methoxy-1H-indol-3-yl)ethyl)acetamide (I-16)

[0092] The 5-norbornene-2-methylamine in Example 2 was replaced by 5-methoxytryptamine, and the other operation steps were the same as those in Example 2. The target compound synthesized was a white solid with a yield of 67.0%.

[0093] 1 H NMR (400mHz, DMSO-d6) δ10.64(s,1H),7.73(t,J=5.6Hz,1H),7.21(d,J=8.7Hz,1H),7.12( d,J=2.4Hz,1H),7.07(d,J=2.4Hz,1H),6.87(d,J=3.5Hz,1H),6.71(dd,J=8.7,2.4Hz,1H) ,6.62(d,J=2.3Hz,1H),5.84(s,1H),3.76(s,3H),3.51-3.42(m,1H),3.33-3.25(m,1H),2 .81(t,J=7.0Hz,2H),2.36(s,3H),1.86(s,3H),1.75(d,J=12.0Hz,3H),1.56-1.42(m,9H). 13 C NMR (100mHz, CDCl3) δ172.2,154.2,140.4,139.8,131.6,127.8,125.5,124.3,123. 0,112.8,112.5,112.1,100.6,82.9,56.1,40.4,39.5,36.8,36.4,28.7,25.3,15.2.

[0094] Example 17. Synthesis of 2-(adamantan-1-yl)-2-hydroxy-2-(5-methylthiophen-2-yl)-N-(2-(5-chloro-1H-indol-3-yl)ethyl)acetamide (I-17)

[0095] The 5-norbornene-2-methylamine in Example 2 was replaced by 5-chlorotryptamine hydrochloride, and the other operation steps were the same as those in Example 2. The target compound synthesized was a white solid with a yield of 78.5%.

[0096] 1 H NMR (400mHz, DMSO-d6) δ11.03 (s, 1H), 7.77 (t, J = 5.2Hz, 1H), 7.63 (s, 1H), 7.34 (d,J=8.6Hz,1H),7.25(s,1H),7.05(d,J=8.6Hz,1H),6.86(d,J=3.5Hz,1H),6.6 1(d,J=3.5Hz,1H),5.83(s,1H),3.51-3.42(m,1H),3.31-3.24(m,1H),2.88-2. 79(m,2H),2.36(s,3H),1.85(s,3H),1.73(d,J=12.0Hz,3H),1.56-1.39(m,9H). 13 C NMR (100mHz, CDCl3) δ172.2,140.3,139.9,134.8,128.6,125.5,125.4,124.4,123. 6,122.6,118.4,113.0,112.3,83.0,40.5,39.6,36.8,36.5,29.9,28.7,25.2,15.2.

[0097] Test Example 1, in vitro anti-influenza virus activity test

[0098] To evaluate the in vitro anti-influenza virus activity of the synthesized compounds, the following virus strains were selected: adamantane-sensitive virus strain A / Hong Kong / 8 / 68 (H3N2) and adamantane-resistant virus strain A / WS / 33 (H1N1). These virus strains were amplified in chicken embryos and their TCID values ​​were determined by the Reed-Muench method. 50 ). Afterwards, the anti-influenza activity of the tested compounds was verified by CPE inhibition assay on infected MDCK cells. Amantadine (AMD) and oseltamivir phosphate (OSV-P) were dissolved in MEM, and the test drugs were dissolved in DMSO and stored at room temperature.

[0099] (1) Antiviral efficacy test

[0100] Centrifuge at 10,000 rpm for 1 minute to remove the drug to the bottom of the tube, then add 100 μL DMSO and dissolve it in MEM containing 1.5 μg / mL TPCK to the concentration to be tested. The final DMSO concentration is 0.5%. Wash the 96-well plate monolayer cells once with PBS and add about 100 TCID50 100 μL / well of the virus dilution was added, incubated in a 37°C, 5% CO2 incubator for 2 hours, the virus solution was discarded, and 2-fold gradient dilutions of the drug were added, with a concentration range of 37.6-1.2 μM. Four replicate wells were set for each concentration, and incubated in a 37°C, 5% CO2 incubator for 48 hours. Observe the cytopathic effect (CPE) under a microscope. If all the cells in the control group have died, it means that the influenza virus has completely infected the cells and caused cytopathic effect. The occurrence of CPE in cells was recorded according to the 6-level standard (as shown in Table 1). Tap the cell culture plate, then carefully remove the culture medium with a pipette, add 1:10 CCK8 detection reagent, 50 μL per well. Then place it in a 37°C incubator for 2 hours, and measure the A450 absorbance value. Use Prism non-regression analysis to calculate the drug's half-effective concentration (EC 50 ), and the results are shown in Table 1.

[0101] (2) Compound toxicity test (MTT method) experimental steps

[0102] Take MDCK cells in good condition and place them in each well at a rate of 2 × 10 4 Inoculate a 96-well plate and incubate at 37°C, 5% CO2 for 24 hours to allow MDCK cells to grow into a monolayer. After 24 hours, aspirate the culture supernatant and rinse the 96-well plate twice with 50 μL / well sterile phosphate-buffered saline (PBS). Then dilute the compound by diluting it 5-fold with serum-free MEM, and add 50 μL / well of the compound to the 96-well plate cells. The cells were cultured in a CO2 incubator at 37°C. After 48 hours, add 5 mg / mL of freshly prepared MTT solution to each well and incubate at 37°C for 4 hours. The dissolution medium was then removed. The absorbance of each well at 490 nm was measured using a CLARIOstar all-purpose multifunctional microplate reader (BMG Labtech, Germany). Formazan crystals were dissolved in DMSO (100 μL per well). Cell viability (%) = OD value of the compound well / average OD value of the control well. TC value of each compound 50 The values ​​were obtained by analyzing the data using a nonlinear regression model using Graph Prism 6 software. The results are shown in Table 1.

[0103] Table 1. In vitro anti-influenza activity data

[0104]

[0105]

[0106] Data are presented as themean±SD of the results of two or three independent tests.a EC50represents halfmaximal effective concentration.b TC50represents 50% cytotoxic concentration,as determined by themTT cell viabilitytest.c SI is the drug selectivity index,the TC50 / EC50 ratio(H1N1).dN.A.,Noactivity

[0107] As shown in Table 1, the α-hydroxycarboxylic acid derivatives with adamantyl provided by the present invention have different degrees of inhibitory activity against wild-type (H3N2) and variant (H1N1) influenza viruses. 50 Among them, compound Ⅰ-16, which has excellent inhibitory activity, has inhibitory activities of 0.92μM and 0.55μM against two virus strains, respectively, which is comparable to the first-line anti-influenza drug oseltamivir, and has the best selectivity index, indicating that the compound has low toxicity and has the potential to be further developed into a new anti-influenza drug.

[0108] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions and improvements made within the spirit and principles of the present invention should be included in the protection scope of the present invention.

Claims

1. An α-hydroxycarboxylic acid derivative having an adamantyl group, a stereoisomer thereof or a pharmaceutically acceptable salt thereof, characterized in that: It has the following structural formula I: Wherein, when X=O in the general formula, n=0 or 1, A is a lipophilic group or a hydrophilic group; In the general formula, when X=N, n=0-3, A is an alkyl group, a cycloalkyl group, a heterocycloalkyl group, an aryl group, a heteroaryl group, a cyclic aromatic group, a heterocyclic aromatic group, a tetrahydropyrrole group, a piperazine group, a piperidine group, a morpholinyl group and other cyclic amines, and one of the substituted derivatives of the above groups.

2. The α-hydroxycarboxylic acid derivative having an adamantyl group according to claim 1, characterized in that Select any one of the following structural formulas:

3. The method for preparing an α-hydroxycarboxylic acid derivative having an adamantyl group according to claim 1 or 2, characterized in that: The synthetic route is: The following steps are involved: Step 1: using 2-adamantyl-α-keto acid 1 as a raw material and cis-N-toluenesulfonylaminoindanol 2 under the action of a condensing agent to generate an ester intermediate 3; Step 2: Compound 3 obtained in step 1 undergoes a nucleophilic addition reaction with 2-methylthiophene lithium prepared in situ at low temperature to obtain compound 5; Step 3, the compound 5 obtained in the above step 2 is hydrolyzed under the conditions of LiOH / CH3OH / THF to obtain a carboxylic acid derivative 6; Step 4: Take the carboxylic acid compound 6 obtained in the above step 3 and condense it with differently substituted primary amines (R-NH2) via a condensing agent to obtain target compounds Ⅰ-1~Ⅰ-4, Ⅰ-7~Ⅰ-17; or react compound 6 with a bromide under NaHCO3 to generate ester compounds Ⅰ-5 and Ⅰ-6.

4. The method for preparing an α-hydroxycarboxylic acid derivative having an adamantyl group according to claim 3, characterized in that: In the step 1, the molar ratio of α-keto acid 1 to the catalyst is 1:05-1.5, the selected catalyst is EDCI·HCl / DMAP or TBTU, and the solvent is dichloromethane or THF.

5. The method for preparing an α-hydroxycarboxylic acid derivative having an adamantyl group according to claim 3, characterized in that: In the step 2, the reaction temperature is -78°C and the reaction time is 3-5h.

6. The method for preparing an α-hydroxycarboxylic acid derivative having an adamantyl group according to claim 3, characterized in that: The solvent ratio selected in step 3 is tetrahydrofuran: methanol: water in a volume ratio of 2:3:1, and the temperature is 60-80°C.

7. The method for preparing an α-hydroxycarboxylic acid derivative having an adamantyl group according to claim 3, characterized in that: The catalyst used to form the amide in step 4 is CDI or EDCI·HCl / HOBt, the solvent is dichloromethane, and the temperature is room temperature 25°C.

8. Use of the α-hydroxycarboxylic acid derivative having an adamantyl group as claimed in claim 1 or 2, and its pharmaceutically acceptable salt, crystalline hydrate and solvate in the preparation of anti-influenza virus drugs.

9. Use of the α-hydroxycarboxylic acid derivative having an adamantyl group and its pharmaceutically acceptable salt, crystalline hydrate and solvate in the preparation of anti-influenza virus drugs according to claim 8, characterized in that: The influenza virus is influenza A H3N2 or H1N1.

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