CN115772117A - 2-hydroxysuccinic acid compound and pharmaceutical composition and application thereof - Google Patents

Abstract

The invention discloses a 2-hydroxysuccinic acid compound, a pharmaceutical composition and an application thereof, wherein the compound is a compound with a structure shown in formula I or II or an isomer, a pharmaceutically acceptable salt or a mixture thereof. The compound and the pharmaceutical composition thereof can effectively inhibit the citric acid transport activity of HEK293 cells, are used for treating metabolic diseases and/or cardiovascular diseases, can exert the drug effect at the molecular level, and have better treatment effectExcellent, and can optimally reach the nanomolar concentration level. In addition, the preparation method of the compound is simple and convenient and is easy to operate.

Description

A kind of 2-hydroxysuccinic acid compound and its pharmaceutical composition and application

technical field

The present invention relates to a compound and its pharmaceutical composition and application, in particular to a 2-hydroxysuccinic acid compound and its pharmaceutical composition and application.

Background technique

Cardiovascular disease is currently the leading cause of death in the world, and dyslipidemia caused by high cholesterol is one of the major risks leading to death from cardiovascular disease. Statins are the first-line treatment for hyperlipidemia, but 15% of patients have clinical defects such as statin intolerance or poor lipid-lowering effect of statins. Therefore, it is of great research significance to develop efficient and safe drugs for the treatment of hyperlipidemia.

Cytosolic citrate is a key precursor and regulator of de novo fatty acid synthesis and is considered an important intermediate linking glucose metabolism and lipid metabolism. The concentration of citrate in the cytoplasm directly affects the rate of fat synthesis. There are three main sources of citrate in the cytoplasm: (1) The cytoplasmic citrate transporter (NaCT) encoded by the SLC13A5 gene takes up citrate in the plasma into the cytoplasm. (2) The mitochondrial inner membrane citrate transporter (PMCT) encoded by the SLC25A1 gene transports excess citrate in the tricarboxylic acid cycle to the cytoplasm. (3) α-ketoglutaric acid (α-KG) is metabolized from glutamine to enter the tricarboxylic acid cycle to generate citric acid. All three sources of citric acid need to generate acetyl-CoA and oxaloacetate under the action of cytoplasmic ATP-citrate lyase (ACLY), and then use acetyl-CoA as the starting material to carry out the de novo lipid synthesis pathway (DNL ). Therefore, inhibiting the key transporters of citrate uptake and the joint enzymes of citrate catabolism are important targets for the treatment of lipid metabolism diseases.

The citrate transporter encoded by the SLC13A5 gene is also known as the plasma membrane citrate transporter/sodium ion-dependent citrate transporter (NaCT). in the pulp. The SLC13A5 gene is highly expressed in the liver of mammals, and also has a certain distribution in the kidney, testis, and brain. In obesity, non-alcoholic steatohepatitis, diabetes and other patients, the expression level of SLC13A5 gene was significantly up-regulated. Mice knocked out of the SLC13A5 gene can avoid the occurrence and development of related metabolic diseases induced by high-fat diet, and NaCT has become an ideal target for regulating energy metabolism and lipid metabolism.

At present, there is no NaCT inhibitor successfully marketed for the treatment of hyperlipidemia, only Pfizer has reported PF-06649298 and PF-06761281, and Bristol-Myers Squibb has reported BI-01383298, PF-06761281 will increase plasma and urine The concentration of citric acid is medium, so the safety needs to be further studied, and the druggability needs to be further improved.

Contents of the invention

Purpose of the invention: the present invention aims to provide a 2-hydroxysuccinic acid compound that inhibits NaCT, which can effectively solve the problems of insufficient NaCT inhibitory activity and poor druggability in existing compounds; another purpose of the present invention is to provide a A pharmaceutical composition with the 2-hydroxysuccinic acid compound as an active ingredient; another object of the present invention is to provide a kind of 2-hydroxysuccinic acid compound in the preparation of Na ⁺ dependent citrate transporter-related diseases application in medicines. Extracellular citrate uptake inhibitors

Technical solution: The 2-hydroxysuccinic acid compound described in the present invention is a compound with formula I or II or its isomer, pharmaceutically acceptable salt or their mixture:

R ₁ is hydrogen, halogen, cyano, C ₁ -C ₄ alkyl, C ₁ -C ₄ haloalkyl, C ₁ -C ₄ alkoxy, C ₁ -C ₄ haloalkoxy, pyrazolyl or phenyl ;

R ₂ is a 6-10 membered aryl group, a 5-10 membered heteroaryl group or a tetrahydroquinolyl group, benzothiazolyl group, or benzoxazolyl group whose 1-4 hydrogens are substituted by R ^2a ;

R ^2a is hydrogen, halogen, cyano, C ₁ -C ₄ alkyl, C ₁ -C ₄ haloalkyl, C ₁ -C ₄ alkoxy or C ₁ -C ₄ haloalkoxy;

L and M are selected from -CH ₂ -, -NH- or -O-;

The heteroatoms in the 5-10 membered heteroaryl group are N, O or S, and the number of heteroatoms is 1-4.

Preferably, in the structure:

_R is hydrogen, fluoro, cyano, methyl, ethyl, propyl, isopropyl, trifluoromethyl, isopropyl, methoxy, ethoxy, isopropoxy, pyrazolyl or benzene base;

R ₂ is phenyl, pyrazolyl, pyridyl or tetrahydroquinolyl, benzothiazolyl, benzoxazolyl substituted by 1-4 hydrogen R ^2a ;

R ^2a is hydrogen, fluoro, cyano, methyl, ethyl, isopropyl, t-butyl, trifluoromethyl, dioxol, methoxy, ethoxy or isopropoxy.

Preferably, the 2-hydroxysuccinic acid compound is selected from any of the following compounds:

Preferably, the pharmaceutically acceptable salt is a salt formed by the above-mentioned compound with an acid or a base, and the acid is hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, methanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid, naphthalene Sulfonic acid, citric acid, tartaric acid, lactic acid, pyruvic acid, acetic acid, maleic acid, succinic acid, fumaric acid, salicylic acid, phenylacetic acid or mandelic acid, the base is containing alkaline metal cations, alkaline earth metal cations or inorganic bases of ammonium cation salts.

The above-mentioned inhibitors and pharmaceutically acceptable carriers form a pharmaceutical composition, which is made into common pharmaceutical preparations, such as tablets, capsules, syrups, suspensions or injections. The preparations can be added with spices, sweeteners, liquid/solid fillers, Common pharmaceutical excipients such as diluents.

The above-mentioned inhibitors and pharmaceutical compositions thereof can be prepared as medicines for treating diseases related to Na ⁺ dependent citrate transporters, specifically for treating hyperlipidemia, diabetes, non-alcoholic steatohepatitis or cancer.

Beneficial effects: Compared with the prior art, the present invention has the following significant advantages: (1) The compound and its pharmaceutical composition can effectively inhibit the activity of NaCT encoded by the SLC13A5 gene, and effectively inhibit the uptake of citric acid by HEK293T cells. Inhibition IC50 value is optimally less than 60nM; (2) This type of compound and its pharmaceutical composition are widely used, and can be prepared as drugs for treating metabolic diseases such as hyperlipidemia, and can exert drug effects at the molecular level, and the therapeutic effect is more excellent , the optimal nanomolar concentration level can be reached; (3) The preparation method of the compound is simple and easy to operate.

Description of drawings

Figure 1 is a graph showing the effect of compound LA-33 on lipid accumulation in AML12 cells;

Figure 2 is a graph showing the effect of compound LA-33 on AML12 cells SLC13A5 and ACLY;

Figure 3 is a graph showing the effect of compound LA-33 on lipid accumulation in primary liver cells of mice;

Figure 4 is a graph showing the effect of compound LA-33 on primary mouse hepatocytes SLC13A5 and ACLY;

Figure 5 is a graph showing the effect of compound LA-33 on plasma lipid levels in starvation-induced mice;

Figure 6 is a graph showing the effect of compound LA-33 on liver lipid accumulation in starvation-induced mice;

Fig. 7 is a graph showing the effect of compound LA-33 on mRNA related to liver lipid accumulation in starvation-induced mice.

Detailed ways

The technical solutions of the present invention will be further described below in conjunction with the examples.

Example 1

Synthesis of LA-1:

Step 1: Add cuprous iodide (0.2g, 1.05mmol) in 60ml THF, add triethylamine (1.13g, 44.32mmol), the reaction solution changes from off-white turbidity to light gray and slightly clear, nitrogen replacement three times, in P-bromophenylacetylene (4g, 22.09mmol) and monoethyl oxalyl chloride (6g, 43.94mmol) were slowly added dropwise under ice-cooling, the reaction solution became pale yellow and then turbid to khaki. The reaction solution was reacted at room temperature for 16 h and then stopped. Add 60ml of saturated sodium bicarbonate solution to the reaction solution, separate the organic layer, extract the aqueous layer three times with ethyl acetate, combine the organic layers, wash twice with saturated brine, dry over anhydrous sodium sulfate, and concentrate under reduced pressure to remove the solvent. Column chromatography using PE:EA=60:1 gave 4 g of light yellow oil. ¹ HNMR(300MHz,DMSO-d ₆ )δ7.62–7.53(m,4H),4.37(s,2H),1.34(s,3H).HR-MS(ESI):Calculated for C ₁₂ H ₁₀ BrO ₃ [M+H] ⁺ :280.9813,found280.9803.

Step 2: Dissolve ethyl acetate (5.01g, 56.9mmol) in THF, cool down to -78°C, add bis(trimethylsilyl)amide lithium (56.9ml, 56.9mmol) dropwise to keep the reaction solution at -78°C After reacting at ℃ for 30 min, 1-1 was dissolved in THF and added dropwise to the reaction solution. After keeping at -78°C for 1.5 h, the reaction was complete as monitored by TLC. The reaction was stopped, quenched with saturated ammonium chloride, extracted with EA, and the organic layers were combined and dried over anhydrous sodium sulfate. 8 g of yellow oil was obtained by column chromatography using PE:EA=15:1. ¹ H NMR (300MHz, Chloroform-d) δ7.52–7.42(m,2H),7.37–7.27(m,2H),4.39(qd,J＝7.1,1.3Hz,2H),4.22–4.15(m, 2H), 3.28(d, J=16.5Hz, 1H), 3.14(d, J=16.5Hz, 1H), 1.38(t, J=7.1Hz, 3H), 1.29(t, J=7.1Hz, 3H) .HR-MS(ESI): Calculated for C ₁₆ H ₁₈ BrO ₅ [M+H] ⁺ :369.0338,found 369.0366.

Step 3: 1-2 (0.50g, 1.35mmol) and 2-methoxypyridine-4-boronic acid pinacol ester (0.36g, 1.62mmol) were dissolved in 7ml solvent (dioxane:water=6 :1), add anhydrous sodium carbonate (0.43g, 4.06mmol), add Pd(dppf)Cl ₂ (0.10g, 0.14mmol) after nitrogen replacement three times, react with 95°C for 5h after nitrogen replacement three times, and then detect the reaction by TLC completely. The reaction solution was cooled to room temperature and filtered with suction, the filtrate was concentrated, ethyl acetate was added, washed twice with water and once with saturated brine, the organic layer was dried over anhydrous sodium sulfate, the solvent was removed under reduced pressure, and a column layer of PE:EA=10:1 was used to 200 mg of a light yellow oily substance was obtained.

¹ H NMR (300MHz, DMSO-d ₆ ) δ8.23 (d, J=5.0Hz, 1H), 7.80–7.70 (m, 2H), 7.61–7.51 (m, 2H), 7.51 (dd, J=5.1 ,1.0Hz,1H),7.08(d,J=1.0Hz,1H),4.31(s,2H),4.10(s,2H),3.89(s,3H),2.82(d,J=0.4Hz,2H ), 1.25 (d, J=15.6Hz, 6H). HR-MS (ESI): Calculated for C ₂₂ H ₂₄ NO ₆ [M+H] ⁺ : 398.1604, found 398.1600.

Step 4: Dissolve intermediates 1-3 in 30ml of absolute ethanol, add 5ml of Raney nickel catalyst, replace with hydrogen three times and react overnight. After the completion of the reaction as detected by TLC, the reaction solution was suction filtered, the filtrate was concentrated, and purified using a silica gel column to obtain 120 mg of a colorless oil. ¹ H NMR (300MHz, DMSO-d ₆ ) δ8.23(d, J=5.0Hz, 1H), 7.58–7.47(m,3H), 7.27–7.16(m,2H), 7.08(d,J=1.0 Hz, 1H), 4.17(d, J=0.5Hz, 2H), 4.09(s, 2H), 3.88(s, 3H), 2.83–2.67(m, 2H), 2.53(dt, J=3.1, 1.0Hz ,2H),2.13(d,J＝0.8Hz,2H),1.22(s,3H),1.14(s,3H).C ₂₂ H ₂₈ NO ₆ [M+H] ⁺ :402.1917,found 402.1933.

Step 5: Dissolve intermediate 1-4 in 5ml of absolute ethanol, add 1.5ml of 1N NaOH solution, react overnight at room temperature, after HPLC detects that the reaction is complete, evaporate the solvent under reduced pressure, and use 1mol/L hydrochloric acid to adjust the pH to 2- 3. The aqueous layer was extracted with ethyl acetate (3×5ml), the organic phases were combined, dried over anhydrous sodium sulfate, and filtered with suction. The filtrate was distilled off the solvent under reduced pressure to obtain 70 mg of a white solid. ¹ H NMR (300MHz, DMSO-d ₆ ) δ8.23(d, J=5.0Hz, 1H), 7.58–7.47(m,3H), 7.27–7.16(m,2H), 7.08(d,J=1.0 Hz,1H),5.14(s,1H),3.88(s,3H),2.82–2.66(m,2H),2.59–2.41(m,2H),2.20–2.02(m,2H).HR-MS( ESI):Calculated for C ₁₈ H ₂₀ NO ₆ [M+H] ⁺ :346.1291,found346.1308.

Using similar operations as in Example 1, the following compounds were obtained:

¹ H NMR (300MHz, DMSO-d ₆ ) δ7.59(d, J=8.1Hz, 2H), 7.37(t, J=7.9Hz, 1H), 7.31–7.04(m, 4H), 6.93(ddd, J＝8.1, 2.6, 1.0Hz, 1H), 3.83(s, 3H), 2.79(t, J＝12.1Hz, 2H), 2.57(d, J＝15.7Hz, 1H), 2.45(dd, J＝13.5 ,5.3Hz,1H),2.01–1.83(m,2H).HR-MS(ESI):Calculated for C ₁₉ H ₂₁ O ₆ [M+H] ⁺ :345.1338,found 345.1372.

¹ H NMR (300MHz, DMSO-d ₆ ) δ7.60–7.50(m,2H),7.40(dt,J＝7.4,2.0Hz,1H),7.35–7.16(m,4H),6.96(dt,J ＝7.4,2.0Hz,1H),5.14(s,1H),4.08(s,2H),2.82–2.66(m,2H),2.59–2.41(m,2H),2.20–2.02(m,2H), 1.40(s,3H).HR-MS(ESI): Calculated for C ₂₀ H ₂₃ O ₆ [M+H] ⁺ :359.1495,found 359.1507

¹ H NMR (300 MHz, DMSO-d ₆ ) δ7.26 (q, J=8.2 Hz, 4H), 6.86 (dd, J=7.4, 1.6 Hz, 1H), 6.76 (dd, J=7.5, 1.7 Hz ,1H),6.52(t,J=7.4 Hz,1H),3.16(t,J=5.5 Hz,2H),2.85–2.69(m,4H),2.58(d,J=15.6 Hz,1H),2.45 (dd,J=13.3,5.4 Hz,1H),2.04–1.74(m,4H).HR-MS(ESI):Calculated for C ₂₁ H ₂₄ NO ₅ [M+H] ⁺ :370.1654,found 370.1660.

¹ H NMR (300 MHz, DMSO-d ₆ ) δ7.50–7.41 (m, 3H), 7.30 (d, J＝7.9 Hz, 2H), 6.37 (d, J＝1.9 Hz, 1H), 3.85 (s ,3H),2.87–2.75(m,2H),2.61(s,1H),1.94(qt,J＝14.0,6.3 Hz,2H).HR-MS(ESI):Calculated for C ₁₆ H ₁₉ N ₂ O ₅ [M+H] ⁺ :319.1294, found 319.1300.

¹ H NMR (300 MHz, DMSO-d ₆ ) δ8.14(s, 1H), 7.82(s, 1H), 7.48(d, J=7.9 Hz, 2H), 7.15(d, J=7.9 Hz, 2H ), 4.15(q, J=7.3 Hz, 2H), 2.80(d, J=15.7 Hz, 1H), 2.73–2.64(m, 1H), 2.59(s, 1H), 2.40(dd, J=13.5, 5.5 Hz,1H),2.01–1.77(m,2H),1.41(t,J＝7.3 Hz,3H).HR-MS(ESI):Calculated for C ₁₇ H ₂₂ N ₂ O ₅ [M+H] ⁺ :333.1450, found 333.1455.

¹ H NMR (300 MHz, DMSO-d ₆ ) δ7.70 (dd, J=6.5, 1.0 Hz, 3H), 7.23–7.13 (m, 2H), 6.82 (s, 1H), 5.14 (s, 1H) ,2.71(d,J＝12.4 Hz,1H),2.59–2.41(m,2H),2.20–2.02(m,2H).HR-MS(ESI):Calculated for C ₁₅ H ₁₇ N ₂ O ₅ [M +H] ⁺ :305.1137, found 305.1144.

¹ H NMR (300 MHz, DMSO-d ₆ ) δ7.80–7.70(m,2H),7.63(s,1H),7.28–7.18(m,2H),6.80(s,1H),5.14(s, 1H), 4.52(s, 1H), 2.71(d, J=12.4 Hz, 1H), 2.59–2.41(m, 2H), 2.20–2.02(m, 2H), 1.44(d, J=15.1 Hz, 5H ).HR-MS(ESI):Calculated for C ₁₈ H ₂₃ N ₂ O ₅ [M+H] ⁺ :347.1607,found 347.1618.

¹ H NMR (300 MHz, Methanol-d ₄ ) δ7.98(s, 1H), 7.58(s, 1H), 7.50(d, J=8.6 Hz, 1H), 7.28(dd, J=8.7, 1.6 Hz ,1H),3.10–2.86(m,2H),2.81–2.56(m,2H),2.09(dtd,J＝26.0,13.5,6.7 Hz,2H).HR-MS(ESI):Calculated for C ₁₃ H ₁₅ N ₂ O ₅ [M+H] ⁺ :279.0981,found 279.0996.

¹ H NMR (300 MHz, DMSO-d ₆ ) δ7.60–7.50(m,2H),7.42–7.30(m,2H),7.27–7.16(m,2H),6.89(d,J＝7.5 Hz, 1H), 6.04(d, J＝0.7 Hz, 2H), 5.14(s, 1H), 2.59–2.41(m, 2H), 2.20–2.02(m, 2H).HR-MS(ESI): Calculated for C ₁₉ H ₁₉ O ₇ [M+H] ⁺ :359.1131,found 359.1144.

¹ H NMR (300 MHz, DMSO-d ₆ ) δ7.50 (h, J = 4.6, 3.8 Hz, 4H), 7.39 (d, J = 7.8 Hz, 2H), 7.28 (d, J = 7.8 Hz, 2H ),2.81(d,J=15.6 Hz,2H),2.60(s,1H),2.44(d,J=5.1 Hz,1H),1.94(dq,J=12.4,6.9,6.0 Hz,2H).HR -MS(ESI):Calculated for C ₁₉ H ₁₉ O ₇ [M+H] ⁺ :399.1055,found 399.1068.

¹ H NMR (300 MHz, DMSO-d ₆ ) δ 7.39 (d, J = 8.0 Hz, 2H), 7.30 (dd, J = 15.3, 7.6 Hz, 2H), 7.20 (d, J = 7.8 Hz, 2H ),7.15–6.97(m,2H),3.76(s,3H),2.87(d,J＝19.9 Hz,1H),2.81–2.69(m,2H),2.60(s,1H),1.93(qd, J＝14.0, 13.0, 5.9 Hz, 2H). HR-MS (ESI): Calculated for C ₁₉ H ₂₁ O ₆ [M+H] ⁺ : 345.1338, found 345.1372.

¹ H NMR (300 MHz, Chloroform-d) δ7.54 (d, J = 7.8 Hz, 2H), 7.37 (d, J = 7.9 Hz, 1H), 7.28 (d, J = 7.8 Hz, 2H), 7.22 –7.10(m, 2H), 6.91(dd, J=8.2, 2.5 Hz, 1H), 4.12(q, J=6.9Hz, 2H), 3.16(d, J=17.0 Hz, 1H), 2.93(q, J＝11.1 Hz, 2H), 2.68(t, J＝12.3 Hz, 1H), 2.15(t, J＝12.0 Hz, 2H), 1.48(d, J＝13.9 Hz, 3H).HR-MS(ESI) :Calculated for C ₂₀ H ₂₃ O ₆ [M+H] ⁺ :359.1495,found 359.1508.

¹ H NMR (300 MHz, DMSO-d ₆ ) δ7.61 (dd, J＝7.5, 2.0 Hz, 1H), 7.54–7.45 (m, 2H), 7.40 (td, J＝7.5, 2.0 Hz, 1H) ,7.25–7.16(m,2H),7.11(td,J=7.5,2.0 Hz,1H),6.95(dd,J=7.5,2.0 Hz,1H),5.14(s,1H),4.68(s,1H ),2.82–2.66(m,2H),2.59–2.41(m,2H),2.20–2.02(m,2H),1.35(d,J＝14.9 Hz,5H).HR-MS(ESI):Calculated for C ₂₁ H ₂₅ O ₆ [M+H] ⁺ :373.1651, found 373.1673.

¹ H NMR (300 MHz, DMSO-d ₆ ) δ7.58 (d, J = 8.2 Hz, 2H), 7.35 (t, J = 7.9 Hz, 1H), 7.29–7.11 (m, 4H), 6.91 (dd ,J=8.2,2.4 Hz,1H),4.00(t,J=6.5 Hz,2H),2.84–2.68(m,2H),2.60(s,1H),2.01–1.81(m,2H),1.75( p, J＝7.1 Hz, 2H), 1.01(t, J＝7.4 Hz, 3H). HR-MS (ESI): Calculated for C ₂₁ H ₂₅ O ₆ [M+H] ⁺ : 373.1651, found 373.1679.

¹ H NMR (300 MHz, DMSO-d ₆ ) δ7.84 (dd, J=7.5, 2.0 Hz, 1H), 7.69 (td, J=7.3, 2.0 Hz, 1H), 7.63 (dd, J=7.5, 2.4 Hz, 1H), 7.58–7.45(m, 3H), 7.25–7.14(m, 2H), 5.14(s, 1H), 2.82–2.66(m, 2H), 2.59–2.41(m, 2H), 2.20 –2.02(m,2H).HR-MS(ESI): Calculated for C ₁₉ H ₁₈ F ₃ O ₅ [M+H] ⁺ :383.1106,found 383.1116.

¹ H NMR (300MHz, DMSO-d ₆ ) δ7.84 (dd, J=7.5, 2.0Hz, 1H), 7.69 (td, J=7.3, 2.0Hz, 1H), 7.63 (dd, J=7.5, 2.4 Hz,1H),7.58–7.45(m,3H),7.25–7.14(m,2H),5.14(s,1H),2.82–2.66(m,2H),2.59–2.41(m,2H),2.20– 2.02(m,2H).HR-MS(ESI):Calculated for C ₁₉ H ₁₈ F ₃ O ₅ [M+H] ⁺ :383.1106,found 383.1119.

Example 2

Synthesis of LA-18:

Synthesis of Intermediate 2-2:

Cuprous iodide (0.34g, 3.57mmol) was added to 60ml THF, and triethylamine (7.22g, 0.071mol) was added, the reaction solution changed from off-white turbidity to light gray and slightly clear, and nitrogen replacement was performed three times. (Triethylsilyl)acetylene (5 g, 0.036 mol) and monoethyl oxalyl chloride (9.7 g, 0.076 mol) were added dropwise, the reaction solution became light yellow and turbid, and the reaction solution was reacted at room temperature for 16 h and then stopped. Add 60ml of saturated sodium bicarbonate solution to the reaction solution, extract three times with ethyl acetate, combine the organic layers, wash twice with saturated brine, dry over anhydrous sodium sulfate, concentrate under reduced pressure to remove the solvent, use PE:EA= 60:1 column chromatography yielded 9 g of light yellow oil. Yield 76%. ¹ H NMR(300MHz,DMSO-d ₆ )δ4.34(s,2H),1.32(s,3H),1.03(s,9H),0.94(s,6H).HR-MS(ESI):Calculated for C ₁₂ H ₂₀ NaO ₃ Si[M+Na] ⁺ :263.1079, found 263.1088.

Synthesis of Intermediate 2-3:

Ethyl acetate (1.12g, 12.69mmol) was dissolved in THF, cooled to -78°C, LiHMDS (12.69ml, 12.69mmol) was added dropwise to the reaction solution and kept at -78°C for 30min. Intermediate 2b (2.70g, 7.93mmol) was dissolved in THF and added dropwise to the reaction solution. After keeping at -78°C for 1.5 h, the reaction was complete as monitored by TLC. The reaction was stopped, quenched with saturated ammonium chloride, extracted with EA, and the organic layers were combined and dried over anhydrous sodium sulfate. Column chromatography using PE:EA=15:1 gave 1.3 g of a yellow oil. Yield 34%. ¹ H NMR (300MHz,DMSO-d ₆ )δ4.10(s,2H),3.69–3.53(m,2H),2.65(s,2H),1.22(s,3H),1.14(s,3H), 1.03(s,9H),0.87(s,6H).HR-MS(ESI):Calculated for C ₁₆ H ₃₀ NaO ₄ Si[M+Na] ⁺ :337.1811,found 337.1816.

Synthesis of intermediates 2-4:

Intermediate 2c (1.3g, 3.95mmol) was dissolved in 15ml of anhydrous ether, and tetrabutylammonium fluoride (5.94ml, 5.94mmol) was added dropwise under ice-cooling. After 1.5h, the reaction was monitored by TLC. Use 10ml of saturated ammonium chloride solution to quench the reaction. After the reaction solution is separated, the aqueous layer is extracted three times with ether. The organic layers are combined and dried over anhydrous sodium sulfate. Oil 0.63g, yield 74%. ¹ H NMR (300MHz, DMSO-d ₆ )δ6.66(s,1H),4.31(s,2H),4.10(s,2H),3.65(s,1H),2.76(d,J＝0.3Hz, 3H), 1.25(d, J＝15.6Hz, 6H).HR-MS(ESI): Calculated for C ₁₀ H ₁₄ NaO ₅ [M+Na] ⁺ :237.0739, found 237.0749.

Synthesis of Intermediates 2-5:

Dissolve 4-(4-iodophenyl)morpholine (0.50g, 1.73mmol) in 5ml tetrahydrofuran, add cuprous iodide (33mg, 0.17mmol), 0.7ml triethylamine, replace with nitrogen three times and add Pd( PPh3)4 (200mg, 0.17mmol) was replaced with nitrogen again. After the temperature of the reaction solution rose to 60°C, intermediate 2d (440mg, 2.07mmol) was added dropwise to the reaction solution. After 1.5h, TLC monitored the reaction to be complete. The reaction solution was cooled to room temperature and filtered with suction, the filtrate was added with water, the aqueous layer was extracted with ethyl acetate, washed twice with saturated brine, dried over anhydrous sodium sulfate and filtered with suction, the solvent was removed under reduced pressure, and the crude product was obtained by column chromatography 0.64 g Pale yellow solid. ¹ H NMR (300MHz,DMSO-d ₆ )δ7.52–7.42(m,2H),7.04–6.94(m,2H),4.31(s,2H),4.10(s,2H),3.74(d,J =3.6Hz, 4H), 3.18(d, J=1.1Hz, 4H), 2.82(d, J=0.4Hz, 2H), 1.25(d, J=15.6Hz, 6H).HR-MS(ESI): Calculated for C ₂₀ H ₂₆ NO ₆ [M+H] ⁺ :376.1760,found376.1772.

Synthesis of intermediates 2-6:

Intermediate 2-5 (0.60g, 1.60mmol) was dissolved in 30ml of solvent (methanol: tetrahydrofuran = 6:1), 3ml of Raney nickel was added, hydrogen was replaced three times, and the reaction was carried out at room temperature overnight. After suction filtration, the solvent was removed under reduced pressure to obtain 0.6 g of off-white solid. ¹ H NMR (300MHz, DMSO-d ₆ )δ7.16–7.05(m,2H),6.87–6.77(m,2H),4.17(d,J＝0.6Hz,2H),4.09(s,2H), 3.74(d, J＝3.6Hz, 4H), 3.19(d, J＝1.7Hz, 4H), 2.83–2.67(m, 2H), 2.59–2.40(m, 2H), 2.13(d, J＝0.8Hz ,2H),1.22(s,3H),1.14(s,3H).HR-MS(ESI):Calculated for C ₂₀ H ₃₀ NO ₆ [M+H] ⁺ :380.2073,found 380.2079.

Synthesis of LA-18:

Intermediate 2-6 (0.50 g, 1.32, mmol) was dissolved in 8 ml of absolute ethanol, 5 ml of 1 mol/L NaOH solution was added, stirred at room temperature for 16 h, and the reaction was monitored by TLC for completeness. The solvent was evaporated under reduced pressure, 1mol/L HCl solution was added to adjust the pH to 2-3, extracted three times with ethyl acetate (3×5ml), the organic layer was combined and washed twice with water, once with saturated brine, dried over anhydrous sodium sulfate and reduced to The solvent was removed by autoclaving to obtain 300 mg of a white solid. ¹ H NMR (300MHz, Deuterium Oxide) δ7.15 (d, J=8.3Hz, 2H), 7.00–6.91 (m, 2H), 3.79 (dd, J=6.2, 3.4Hz, 4H), 3.07–2.98 ( m, 4H), 2.68–2.52(m, 2H), 2.41–2.21(m, 2H), 1.76(dtd, J＝41.7, 13.3, 4.8Hz, 2H). HR-MS (ESI): Calculated for C ₁₆ H ₂₂ NO ₆ [M+H] ⁺ :324.1447, found 324.1459.

Using similar operations as in Example 2, the following compounds were obtained:

¹ H NMR (300MHz, DMSO-d ₆ )δ7.10(t, J=0.5Hz, 4H), 5.14(s, 1H), 2.82–2.63(m, 4H), 2.55–2.46(m, 2H), 2.20–2.02(m,2H),1.23(s,3H).HR-MS(ESI):Calculated for C ₁₄ H ₁₉ O ₅ [M+H] ⁺ :267.1232,found 267.1246.

¹ H NMR (300MHz, DMSO-d ₆ ) δ7.10 (tdt, J=7.7, 6.9, 0.9Hz, 4H), 5.14(s, 1H), 2.90 (t, J=0.9Hz, 1H), 2.74( d,J=5.5Hz,2H),2.59–2.42(m,2H),2.20–2.02(m,2H),1.25(d,J=15.1Hz,6H).HR-MS(ESI):Calculated for C ₁₅ H ₂₁ O ₅ [M+H] ⁺ :281.1389, found 281.1366.

¹ H NMR (300MHz,DMSO-d ₆ )δ7.70–7.60(m,2H),7.49–7.39(m,2H),5.14(s,1H),2.82–2.66(m,2H),2.59–2.41 (m,2H),2.20–2.02(m,2H).HR-MS(ESI):Calculated for C ₁₃ H ₁₄ F ₃ O ₅ [M+H] ⁺ :307.0793,found 307.0801.

¹ H NMR (300 MHz, DMSO-d ₆ ) δ7.26–7.09(m, 4H), 5.14(s, 1H), 2.77(d, J=12.4 Hz, 1H), 2.71(d, J=12.4 Hz ,1H),2.59–2.46(m,2H),2.46(dt,J＝7.7,1.0 Hz,1H),2.20–2.02(m,2H),1.73(dd,J＝16.9,13.0 Hz,4H), 1.59 (dd, J＝13.0, 10.1 Hz, 4H). HR-MS (ESI): Calculated for C ₁₇ H ₂₃ O ₅ [M+H] ⁺ : 307.1575, found 307.1554.

¹ H NMR (300 MHz, Chloroform-d) δ7.16–7.06(m,2H),6.84–6.74(m,2H),3.43–3.29(m,4H),2.89–2.71(m,2H),2.59 (dt,J=12.4,1.0 Hz,1H),2.51(dt,J=12.4,1.0 Hz,1H),2.16(d,J=12.4 Hz,1H),2.08–1.98(m,3H).HR- MS(ESI): Calculated for C ₁₆ H ₂₂ NO ₅ [M+H] ⁺ : 308.1498, found 308.1495.

¹ H NMR (300 MHz, Chloroform-d) δ7.26–7.09 (m, 4H), 2.89–2.71 (m, 2H), 2.61–2.42 (m, 3H), 2.19 (d, J=12.3 Hz, 1H ),2.08(s,1H),1.80(d,J=13.1 Hz,2H),1.70(d,J=13.0 Hz,2H),1.61–1.31(m,6H).HR-MS(ESI):Calculated for C ₁₈ H ₂₅ O ₅ [M+H] ⁺ :321.1702, found 321.1718.

¹ H NMR (300 MHz, Chloroform-d) δ7.16–7.06 (m, 2H), 6.84–6.74 (m, 2H), 3.22 (d, J=4.8 Hz, 4H), 2.98 (d, J=9.7 Hz, 4H), 2.89–2.71(m, 2H), 2.55(qt, J=12.4, 1.0 Hz, 2H), 2.16(d, J=12.4 Hz, 1H), 2.03(d, J=12.4 Hz, 1H ).HR-MS(ESI): Calculated for C ₁₆ H ₂₃ N ₂ O ₅ [M+H] ⁺ :323.1607,found 323.1608.

¹ H NMR (300 MHz, Chloroform-d) δ7.17–7.06 (m, 2H), 6.84–6.74 (m, 2H), 3.19 (d, J=13.0 Hz, 4H), 2.84 (d, J=12.4 Hz,1H),2.76(d,J=12.4Hz,1H),2.56(dd,J=12.0,3.8Hz,7H),2.29(s,3H),2.16(d,J=12.4Hz,1H), 2.03 (d, J＝12.4 Hz, 1H). HR-MS (ESI): Calculated for C ₁₇ H ₂₅ N ₂ O ₅ [M+H] ⁺ : 337.1763, found 337.1766.

¹ H NMR (300 MHz, Chloroform-d) δ7.15–7.05(m,2H),6.86–6.76(m,2H),3.78(s,3H),2.89–2.71(m,2H),2.55(qt ,J=12.4,1.0 Hz,2H),2.16(d,J=12.4 Hz,1H),2.03(d,J=12.4 Hz,1H).HR-MS(ESI):Calculated for C ₁₃ H ₁₇ O ₆ [M+H] ⁺ :269.1025, found 269.1033.

¹ H NMR (300 MHz, Chloroform-d) δ7.17–7.06 (m, 2H), 6.85–6.74 (m, 2H), 4.08 (s, 2H), 2.84 (d, J=12.4 Hz, 1H), 2.76(d, J=12.4 Hz, 1H), 2.55(qt, J=12.4, 1.0 Hz, 2H), 2.16(d, J=12.4 Hz, 1H), 2.03(d, J=12.4 Hz, 1H), 1.42(s,3H).HR-MS(ESI): Calculated for C ₁₄ H ₁₉ O ₆ [M+H] ⁺ :283.1182,found 283.1189.

¹ H NMR (300 MHz, Chloroform-d) δ7.15 (d, J = 7.4 Hz, 1H), 6.94 (dtt, J = 7.5, 2.0, 1.0 Hz, 1H), 6.88–6.76 (m, 2H), 3.81(s,3H),2.89–2.71(m,2H),2.57(dt,J＝12.4,1.0Hz,1H),2.47(dt,J＝12.4,1.0Hz,1H),2.16(d,J＝ 12.4 Hz, 1H), 2.04 (d, J＝12.4 Hz, 1H). HR-MS (ESI): Calculated for C ₁₃ H ₁₇ O ₆ [M+H] ⁺ : 269.1025, found 269.1033.

¹ H NMR (300 MHz, DMSO-d ₆ ) δ7.18 (t, J=7.4 Hz, 1H), 6.95 (dtt, J=7.6, 2.0, 1.0Hz, 1H), 6.88–6.75 (m, 2H) ,4.07(s,2H),2.82–2.66(m,2H),2.56(dt,J=12.4,1.0 Hz,1H),2.47(dt,J=12.3,1.0 Hz,1H),2.10(d,J ＝12.4 Hz, 1H), 2.00(d, J＝12.4 Hz, 1H), 1.39(s, 3H).HR-MS(ESI): Calculated for C ₁₄ H ₁₉ O ₆ [M+H] ⁺ :283.1182, found 283.1185.

¹ H NMR (300 MHz, DMSO-d ₆ ) δ8.14(d, J=7.9 Hz, 1H), 8.03(dd, J=11.4, 8.0 Hz, 3H), 7.55(t, J=7.6 Hz, 1H ), 7.46(t, J=7.5 Hz, 1H), 7.38(d, J=7.9 Hz, 2H), 2.81(s, 1H), 2.76(s, 1H), 2.58(d, J=15.5 Hz, 2H ), 1.95 (dq, J＝12.4, 7.0, 6.2 Hz, 2H). HR-MS (ESI): Calculated for C ₁₉ H ₁₈ NO ₅ S[M+H] ⁺ : 372.0906, found 372.0916.

¹ H NMR (300MHz, DMSO-d ₆ ) δ8.14(d, J=7.9Hz, 1H), 8.01(dd, J=11.4, 8.0Hz, 3H), 7.55(t, J=7.6Hz, 1H) ,7.48(t,J=7.5Hz,1H),7.38(d,J=7.9Hz,2H),2.81(s,1H),2.76(s,1H),2.58(d,J=15.5Hz,2H) ,1.95(dq,J＝12.4,7.0,6.2Hz,2H).HR-MS(ESI):Calculated for C ₁₉ H ₁₈ NO ₆ [M+H] ⁺ :356.1134,found 356.1155.

¹ H NMR (300MHz, DMSO-d ₆ ) δ7.69–7.54(m, 4H), 7.46(t, J=7.6Hz, 2H), 7.36(dd, J=8.4, 6.2Hz, 1H), 7.27( d,J=8.1Hz,2H),2.82(d,J=15.7Hz,2H),2.61(s,1H),2.51–2.40(m,1H),2.05–1.81(m,2H)..HR- MS(ESI):Calculated for C ₁₈ H ₁₉ O ₅ [M+H] ⁺ :315.1232,found315.1239.

¹ H NMR (300MHz, DMSO-d ₆ )δ7.89(d, J=5.1Hz, 3H), 7.50(d, J=5.1Hz, 3H), 7.28(d, J=0.6Hz, 1H), 2.78 –2.54(m,5H), 2.11(d, J＝12.4Hz, 1H), 2.00(d, J＝12.3Hz, 1H).HR-MS(ESI): Calculated for C ₁₆ H ₁₇ O ₅ [M+ H] ⁺ :289.1076, found 289.1077.

Example 3

Some compounds of the present invention inhibit the uptake of extracellular D4-citrate in HEK-293T cells with high expression of SLC13A5:

1. Experimental method

(1) Cell recovery and culture: Thaw HEK293T cells taken out of liquid nitrogen quickly in a 37°C water bath for about 1 min, then transfer to a centrifuge tube containing 5 mL of medium, centrifuge at 1100 rpm for 3 min, discard the upper Add 2 mL of 10% FBS DMEM high-glucose medium to resuspend, inoculate 1 mL into a cell culture dish containing 7 mL of medium, put it in a cell culture incubator with 5% CO2 at 37 ° C, and the cells grow in a single layer of adherent. When the cells are confluent to about 90%, discard the medium, wash twice with 37°C preheated PBS, add 2 mL of 0.25% trypsin to digest at room temperature, shake gently for 30 seconds, discard the trypsin, and add 2 mL of fresh medium to terminate the reaction , pipette gently and repeatedly, transfer the medium containing the cells to a sterilized Ep tube, centrifuge at 800rpm for 3min, discard the supernatant after centrifugation, add 2mL of 10% FBS DMEM high glucose medium to resuspend, Inoculated into cell culture dishes, put into 5% CO ₂ , 37°C cell culture incubator for culture.

(2) Transfection: Discard the original medium and replace it with double-antibody-free serum-free high-glucose DMEM medium, and use Opti-MEM serum-free medium to dilute the SLC13A5 overexpression plasmid and Lipofectamine ^TM 2000 transfection reagent in sequence. Add the overexpression plasmid and Lipofectamine ^TM 2000 into a 24-well cell culture plate, shake gently to mix, and place in a cell culture incubator;

(3) Uptake: 24 hours after successful transfection, suck out the original medium in the 24-well plate, add 1mL sodium buffer to each well to wash three times, then add 250μL sodium buffer containing different concentrations of compounds, put it in a shaking incubator, and keep the uptake temperature Incubate at 37°C for 30 minutes; suck out the sodium buffer containing the compound, add 1 mL of sodium buffer to wash three times; add 250 μL of sodium buffer containing the above-mentioned different concentrations of the compound and 200 μM D4-citric acid to each well, and ingest at 37°C for 30 minutes; discard For sodium buffer containing D4-citric acid, add 1 mL choline buffer to each well and wash three times to stop the uptake; after the reaction, add 200 μL double distilled water to each well, place in -80°C for 30 minutes, thaw at room temperature, and freeze and thaw repeatedly for 3 Once, sonicate for 10 minutes to fully break the cells, transfer the solution containing the broken cells to the Ep tube, add acetonitrile containing the internal standard, 12000 rpm, centrifuge for 10 minutes, take the supernatant and use LC-MS/MS to detect the intracellular D4-citric acid concentration, At the same time, subtract the intracellular D4-citrate concentration in the HEK-293T-vector group to calculate the net content.

2. Data processing

(1) Calculation formula for citric acid intake experiment

%Inhibition=[1-(A_sample/A_max)]

Wherein: A_sample represents the D4-citrate content in the sample, and A_max represents the D4-citrate content in the blank.

(2) Fit the dose-effect curve with the log value of the concentration as the X-axis, and the percentage inhibition rate as the Y-axis, and use the log (inhibitor) vs. response-Variable slope of the analysis software GraphPad Prism5 to fit the dose-effect curve, thereby obtaining each _IC50 values of compounds for citric acid uptake activity.

Calculation formula:

Y=Bottom+(Top-Bottom)/(1+10^((Log IC ₅₀ -X)×Hill Slope)).

See Table 1 for _IC50 data.

Table 1. Inhibitory activity of compounds on citric acid uptake in HEK293 cells

serial number Citric acid uptake inhibitory activity serial number Citric acid uptake inhibitory activity LA-1 A LA-2 A LA-3 A LA-4 A LA-5 A LA-6 A LA-7 A LA-8 A LA-9 B LA-10 C LA-11 A LA-12 B LA-13 A LA-14 A LA-15 A LA-16 A LA-17 A LA-18 A LA-19 B LA-20 A LA-21 B LA-22 A LA-23 C LA-24 A LA-25 A LA-26 A LA-27 B LA-28 C LA-29 A LA-30 B LA-31 B LA-32 C LA-33 C LA-34 C

Note: A: <1 μM, B: 1-5 μM, C: >5 μM.

As shown in Table 1, all test compounds have good inhibitory effect on the citrate transporter of HEK293 cells, and all compounds have micromolar inhibition rate on NaCT, among them, compound LA-33 has an inhibitory effect on citrate uptake in HEK293T cells Optimal, IC ₅₀ =60 nM.

Example 4

In vitro drug efficacy test of representative compound LA-33 of the present invention on AML12 cells

Experimental steps:

AML12 cells were cultured in DMEM/F12 high glucose medium containing 10% FBS and placed in a cell culture incubator at 37°C with 5% CO ₂ . The lipid accumulation model was developed by first preprotecting with serum-free medium containing different concentrations of compounds for 30 min and then stimulating hepatocytes with a mixture of PA and OA (designated as OPA, PA/OA, 1:4) at 300 μM for 24 h. To mimic the physiological state of the endogenous extracellular matrix, aliquots of 200 μM citrate were supplemented to the medium. AML12 cells were incubated with OPA for 24 hours, and compound LA-33 with different concentrations were added to the medium for 24 hours to evaluate lipid accumulation and mRNA expression.

As shown in Figure 1, compound LA-33 effectively reduced the content of TC and TG in OPA-stimulated AML cells at a concentration of 10 μM. Figure 2 shows that ACLY is significantly down-regulated, and compound LA-33 only inhibits the function of SLC13A5 without affecting its expression.

Example 5

In vitro pharmacodynamic experiment of representative compound LA-33 of the present invention on mouse primary liver cells

Experimental steps:

Mouse liver primary cells were cultured in DMEM high glucose medium containing 10% FBS and placed in a cell culture incubator at 37°C with 5% _CO2 . The lipid accumulation model was developed by first preprotecting with serum-free medium containing different concentrations of compounds for 30 min and then stimulating hepatocytes with a mixture of PA and OA (designated as OPA, PA/OA, 1:4) at 300 μM for 24 h. To mimic the physiological state of the endogenous extracellular matrix, aliquots of 200 μM citrate were supplemented to the medium. AML12 cells were incubated with OPA for 24 h, and compound LA-33 with different concentrations were added to the medium for 24 h to evaluate lipid accumulation and mRNA expression.

As shown in Figure 3, compound LA-33 effectively reduced the content of TC and TG in OPA-stimulated AML cells. Figure 4 shows that ACLY is significantly down-regulated, and compound LA-33 only inhibits the function of SLC13A5 without affecting its expression, showing the same results as the AML12 cell line.

Example 6

In vivo efficacy test of representative compound LA-33 of the present invention

1. Experimental steps

C57BL/6J mice were purchased from Jiangsu Ai Lingfei Biotechnology Co., Ltd., and after adaptive feeding for 1 week, they were randomly divided into four groups. The blank group was fed normally, and the other three groups were fasted for 48 hours. After fasting, the low-dose group and the high-dose group were injected intraperitoneally with a dose of 10 mg/kg and a dose of 30 mg/kg, respectively. After 1.5 h the animals were anesthetized with sodium pentobarbital (50 mg/kg) intraperitoneally to collect blood samples or liver tissue. Use free fatty acid kit to measure fatty acid (NEFA) in plasma and liver; use triglyceride test kit to measure triglyceride (TG) content in plasma and liver; use total cholesterol test kit to measure triglyceride (TC) in plasma and liver ) content; HDL-c and LDL-c in plasma were measured using low-density lipoprotein cholesterol test kit and high-density lipoprotein cholesterol test kit respectively. Plasma samples were directly measured, and tissue samples were homogenized with 9 times the volume of normal saline for determination.

2. Data processing:

Plasma sample calculation formula:

Cholesterol (TC) content (mmol/L) = (A _sample - A _blank ) / (A _standard - A _blank ) × C _standard

Triglyceride (TG) content (mmol/L) = (A _sample - A _blank ) / (A _standard - A _blank ) × C _standard

NEFA content (mmol/L) = (ΔA _sample - ΔA _blank ) / (ΔA _standard - ΔA _blank ) × C _sample ;

LDL-c and HDL-c content (mmol/L) = (ΔA sample - ΔA blank) / (ΔA standard - ΔA blank) × C sample;

As shown in Figure 5, compound LA-33 can significantly reduce the content of total cholesterol, total triglycerides and free fatty acids in mouse plasma in a dose-dependent manner, and at the same time at a dose of 30 mg/kg can significantly reduce the plasma content of starvation-induced mice. LDL content.

As shown in Figure 6, compound LA-33 can significantly reduce the contents of total cholesterol, total triglycerides and free fatty acids in mouse liver in a significant dose-dependent manner.

As shown in Figure 7, the compound LA-33 can significantly reduce the mRNA expression of ACLY and the mRNA expression of ACC1 and FASN in the DNL pathway in the mouse liver, but the mRNA expression of the key enzyme HMGCR in the TC synthesis pathway has no significant change. It was consistent with the results of liver TC and TG determination.

Claims (7)

1. A 2-hydroxysuccinic compound, which is a compound having a structure represented by formula i or ii or an isomer, a pharmaceutically acceptable salt or a mixture thereof:

R ₁ is hydrogen, halogen, cyano, C ₁ -C ₄ Alkyl radical, C ₁ -C ₄ Haloalkyl, C ₁ -C ₄ Alkoxy radical, C ₁ -C ₄ Haloalkoxy, pyrazolyl or phenyl;

R ₂ is 1-4 hydrogens by R ^2a Substituted 6-10 membered aryl, 5-10 membered heteroaryl, tetrahydroquinolinyl, benzothiazolyl, or benzoxazolyl;

R ^2a is hydrogen, halogen, cyano, C ₁ -C ₄ Alkyl radical, C ₁ -C ₄ Haloalkyl, C ₁ -C ₄ Alkoxy or C ₁ -C ₄ A haloalkoxy group;

l and M are selected from-CH ₂ -, -NH-or-O-;

the heteroatom in the 5-10 membered heteroaryl is N, O or S, and the number of the heteroatoms is 1-4.

2. The compound of claim 1, wherein in the structure:

R ₁ is hydrogen,Fluoro, cyano, methyl, ethyl, propyl, isopropyl, trifluoromethyl, isopropyl, methoxy, ethoxy, isopropoxy, pyrazolyl or phenyl;

R ₂ is 1-4 hydrogen R ^2a Substituted phenyl, pyrazolyl, pyridyl or tetrahydroquinolinyl, benzothiazolyl, benzoxazolyl;

R ^2a is hydrogen, fluorine, cyano, methyl, ethyl, isopropyl, tert-butyl, trifluoromethyl, dioxacene, methoxy, ethoxy or isopropoxy.

3. The compound of claim 1, selected from any one of the following:

LA-1

LA-18

LA-2

LA-19

LA-3

LA-20

LA-4

LA-21

LA-5

LA-22

LA-6

LA-23

LA-7

LA-24

LA-8

LA-25

LA-9

LA-26

LA-10

LA-27

LA-11

LA-28

LA-12

LA-29

LA-13

LA-30

LA-14

LA-31

LA-15

LA-32

LA-16

LA-33

LA-17

LA-34

4. the compound of claim 1, wherein the pharmaceutically acceptable salt is a salt of the compound with an acid or a base, wherein the acid is hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, methanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid, naphthalenesulfonic acid, citric acid, tartaric acid, lactic acid, pyruvic acid, acetic acid, maleic acid, succinic acid, fumaric acid, salicylic acid, phenylacetic acid, or mandelic acid, and the base is an inorganic base comprising an alkali metal cation, an alkaline earth metal cation, or an ammonium cation salt.

5. A pharmaceutical composition comprising a compound according to any one of claims 1 to 4 as an active ingredient, said pharmaceutical composition comprising a pharmaceutically acceptable carrier.

6. Use of a compound according to any one of claims 1 to 4 or a pharmaceutical composition according to claim 5 in the preparation of Na ⁺ Use in the manufacture of a medicament for the treatment of a disease associated with a dependent citrate transporter.

7. Use according to claim 6, wherein said Na ⁺ The dependent citrate transporter-related disease is a metabolic disease or cancer, and specifically is hyperlipidemia or non-alcoholic fatty liver disease.

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