Quaternary ammonium salts of 3-hydroxyisonicotinic acid dextranol ester and their pharmaceutical uses

Abstract

This invention discloses a quaternary ammonium salt compound of 3-hydroxyisonicotinic acid dextran ester with the structure shown in Formula I, wherein R is selected from saturated or unsaturated C1-C6 hydrocarbon groups, substituted or unsubstituted benzyl groups, and the substituent of the benzyl group is selected from halogens, C1-C3 alkyl groups, C1-C3 alkoxy groups, C1-C3 halogen-substituted alkyl groups; R1 is selected from NH2, C1-C4 alkyl groups, C1-C4 alkoxy groups, phenyl groups, and phenoxy groups; R2 is selected from H and C1-C3 alkyl groups; and X is selected from bromine and iodine. The compound of this invention has good anti-inflammatory effects and can be used to prepare drugs for treating inflammation-related diseases. This invention also discloses the use of the quaternary ammonium salt compound of 3-hydroxyisonicotinic acid dextran ester in the preparation of drugs for treating inflammation-related diseases.

Description

Technical Field

[0001] This invention belongs to the pharmaceutical field and relates to a class of quaternary ammonium salt compounds of 3-hydroxyisonicotinic acid dextranol ester and their pharmaceutical uses. Background Technology

[0002] Macrophages play a crucial role in initiating, maintaining, and resolving inflammatory responses. Lipopolysaccharide (LPS), a major component of the cell wall of Gram-negative bacteria, possesses potent immunostimulatory capabilities. RAW264.7 macrophages, as murine immune cells, are activated upon stimulation by external factors (such as LPS), secreting numerous inflammatory factors (e.g., NO, IL-1β, TNF-α) and generating an inflammatory response. The amount of inflammatory factors indirectly reflects the severity of inflammation, serving as a quantitative indicator of its intensity. Summary of the Invention

[0003] In previous research, the inventors discovered a class of derivatives of 2-hydroxynicotinic acid dextran ester or benzoyl ester (CN118206481A). These compounds exhibit good neuroprotective effects and show promising therapeutic effects on central nervous system inflammatory diseases such as stroke, neuropathic pain, and depression. Further research revealed that structural modification of these compounds to obtain 3-hydroxyisonicotinic acid dextran ester derivatives (control compound, compound 1a) did not show significant anti-inflammatory effects. Unexpectedly, the inventors discovered that the quaternary ammonium salt of 3-hydroxyisonicotinic acid dextran ester exhibited a significant anti-inflammatory effect. Simultaneously, the inventors also found that these compounds have a short half-life, allowing them to exert anti-inflammatory effects after local administration. They are also rapidly metabolized in the bloodstream, resulting in better safety. Therefore, they are more suitable for local administration, such as formulations as sprays for respiratory inflammation and eye drops for ocular inflammation. Local administration may achieve better efficacy and safety, leading to its widespread clinical application. For example, corticosteroids have good anti-inflammatory effects, but they are accompanied by toxicity during treatment. Cloteprenol, when used in the eyes, is rapidly metabolized into inactive products, reducing systemic toxicity, while its anti-inflammatory effect is stronger than prednisolone. Budesonide, when inhaled as a 1%-5% solution via nebulization, can reach the whole lungs, inhibiting airway hyperresponsiveness, reducing glandular secretion, promoting airway dilation, reducing airway mucus secretion, and improving lung ventilation. Inhalation therapy directly targets the organs, has a rapid onset of action, minimal side effects, and high safety.

[0004]

[0005] The purpose of this invention is to provide a class of quaternary ammonium salt compounds of 3-hydroxyisonicotinic acid dextran ester.

[0006] The objective of this invention is achieved through the following technical solution:

[0007] Quaternary ammonium salts of 3-hydroxyisonicotinic acid dextran ester with the structure shown in Formula I:

[0008]

[0009] Wherein, R is selected from saturated or unsaturated C1-C6 hydrocarbon groups, substituted or unsubstituted benzyl groups, The substituents of the benzyl group are selected from halogens, C1-C3 alkyl groups, C1-C3 alkoxy groups, and C1-C3 halogen-substituted alkyl groups. R1 is selected from NH2, C1-C4 alkyl, C1-C4 alkoxy, phenyl, phenoxy; R2 is selected from H, C1-C3 alkyl; X is selected from bromine, iodine.

[0010] Preferably, R is selected from substituted or unsubstituted benzyl groups. C1-C3 alkyl groups The substituents of benzyl are selected from fluorine, methyl, methoxy, trifluoromethane, The substituents can be located at the ortho, meta, or para position of the existing substituents on the benzene ring; R1 is selected from NH2, C1-C4 alkyl, C1-C4 alkoxy, phenyl, and phenoxy; R2 is selected from H and methyl; R3, R4, and R5 are each independently selected from H and methyl; X = bromine or iodine.

[0011] More preferably, R is selected from substituted or unsubstituted benzyl groups. The substituents of benzyl are selected from fluorine, methyl, methoxy, and trifluoromethane, and the substituents are located at the para position of the existing substituents on the benzene ring; R3 = R4 = R5 = H; R3 = methyl, R4 = R5 = H; R3 = H, R4 = R5 = methyl; X = bromine, iodine.

[0012] Specifically, the quaternary ammonium salts of 3-hydroxyisonicotinic acid dextran ester with the following structure:

[0013]

[0014]

[0015] Another object of the present invention is to provide the use of the quaternary ammonium salt compound of the aforementioned 3-hydroxyisonicotinic acid dextranol ester in the preparation of medicaments for treating inflammation-related diseases.

[0016] The inflammation-related diseases mentioned include rheumatoid arthritis, eye inflammation, and respiratory tract inflammation.

[0017] Specifically, the eye inflammation mentioned is conjunctivitis.

[0018] Another object of the present invention is to provide a pharmaceutical composition for treating inflammation-related diseases, wherein the pharmaceutical composition comprises, as the active ingredient, a quaternary ammonium salt compound of the aforementioned 3-hydroxyisonicotinic acid dextran ester, and is formulated into a pharmaceutically acceptable dosage form with pharmaceutically acceptable excipients.

[0019] Preferably, the dosage form is a solution, spray, patch, gel, etc.

[0020] Another object of the present invention is to provide the use of the pharmaceutical composition in the preparation of a medicament for treating inflammation-related diseases.

[0021] The beneficial effects of this invention are:

[0022] The quaternary ammonium salts of 3-hydroxyisonicotinic acid dextran ester of this invention have good anti-inflammatory effects and can be used to prepare drugs for treating inflammation-related diseases. Because the quaternary ammonium salts of 3-hydroxyisonicotinic acid dextran ester contain a quaternary ammonium salt structure, these compounds are less likely to enter the central nervous system, thus exhibiting better safety for peripheral inflammation-related diseases. Attached Figure Description

[0023] Figure 1 This invention demonstrates the inhibitory effect of the compound on carrageenan-induced paw edema in rats. Detailed Implementation

[0024] The following embodiments are intended to enable those skilled in the art to fully understand the present invention, but do not limit the invention in any way.

[0025] Example 1

[0026] Synthesis of the target compound

[0027] 1.1) Synthesis of 1-methyl-3-hydroxyisonicotinic acid dextran iodide salt (compound 1)

[0028] Synthesis route:

[0029]

[0030] Step (1): Isonicotinic acid (5.00 mmol), dextroborneol (5.50 mmol), dicyclohexylcarbodiimide (7.50 mmol) and 4-dimethylaminopyridine (2.50 mmol) were reacted in dichloromethane (8 mL) at room temperature for 12 h. After the reaction was completed, the mixture was filtered. The filtrate was collected, evaporated to dryness, dissolved in 100 mL of ethyl acetate, and washed three times with 100 mL of saturated brine each time. The organic layer was collected, dried, and purified by normal-phase silica gel chromatography (petroleum ether: ethyl acetate = 20:1 V / V) to obtain compound 1a (control compound), a white powder with a yield of 73%.

[0031] Step (2): Dissolve compound 1a (1.00 mmol) in 5 mL of acetonitrile, add iodomethane (1.00 mmol), react at 80 °C for 6 h. After the reaction is complete, evaporate to dryness, dissolve in 100 mL of ethyl acetate, wash three times with 100 mL of saturated brine, collect the organic layer, dry it, and then purify it by normal phase silica gel chromatography (petroleum ether: ethyl acetate = 10:1 V / V) to obtain compound 1, a yellow powder, with a yield of 69%.

[0032] 1 H NMR (400MHz, DMSO-d6) (ppm): δ7.03(t,1H),6.28(d,2H),5.01-4.07(m,1H),2.34-2.26(m,1H),2.0 4-1.98(m,1H),1.71-1.63(m,2H),1.29-1.10(m,2H),1.05-1.03(m,1H),0.87(s,3H),0.82(s,6H). 1 H NMR(400MHz, DMSO-d6)δ8.56-8.45(m,2H),8.17(d,J=6.0Hz,1H),5.05(dt,J=9.8,2.9Hz,1H),4.28(s,3H),2.36(ddt,J=13.8,10.0, 4.0Hz,1H),1.93(dt,J=13.0,4.9Hz,1H),1.70(hept,J=4.1Hz,2H),1.34-1.17(m,2H),1.14-1.07(m,1H),0.89(s,3H),0.84(s,6H).

[0033] 13 C NMR (101MHz, DMSO-d6) δ163.77,155.20,136.86,132.04,128.00,82.51,49.35,48.86,48.15,44.74,36.55,28.02,27.27,20.03,19.16,13.93.

[0034] 1.2) Synthesis of 1-(2,3-allyl)-3-hydroxyisonicotinic acid dextran ester bromide (compound 2)

[0035] Following the synthesis method of compound 1, isonicotinic acid dextranol ester and 3-bromopropene were used as raw materials. In step (2) of compound 1, iodomethane was replaced with an equimolar amount of 3-bromopropene. All other steps were the same as in step (2) of compound 1 to synthesize compound 2.

[0036]

[0037] Yellow solid;1 H NMR(400MHz,Chloroform-d)δ9.20(s,1H),9.09(d,J=6.2Hz,1H),8.23(d,J=6.3Hz,1H),6.14(ddt,J =16.7,9.9,6.7Hz,1H),5.72(d,J=16.9Hz,1H),5.62(d,J=6.7Hz,2H),5.56(d,J=10.1Hz,1H),5.16( d,J=9.8Hz,1H),2.46(ddt,J=14.2,9.3,4.1Hz,1H),1.96(m,J=13.6,9.3,4.1Hz,1H),1.79(dq,J=14 .9,4.2Hz,2H),1.45-1.27(m,2H),1.14(dd,J=14.1,3.4Hz,1H),0.93(s,3H),0.89(d,J=4.2Hz,6H).

[0038] 13 C NMR(101MHz,Chloroform-d)δ165.20,158.22,136.39,135.66,129.49,128.26,127.99 ,125.35,84.86,64.32,49.39,48.22,44.86,36.63,27.99,27.31,19.71,18.91,13.67.

[0039] 1.3) Synthesis of 1-(2-methyl-2-allyl)-3-hydroxyisonicotinic acid dextran ester bromide (compound 3)

[0040] Referring to the synthesis method of compound 1, using isonicotinic acid dextranol ester and 2-methyl-3-bromopropene as raw materials, iodomethane in step (2) of compound 1 was replaced with an equimolar amount of 2-methyl-3-bromopropene, and the rest were the same as step (2) of compound 1, to synthesize compound 3.

[0041]

[0042] Yellow solid; 1H NMR(400MHz,Chloroform-d)δ9.14(s,1H),9.05(d,J=6.2Hz,1H),8.24(d,J=6.0Hz,1H),5.55(s,2H),5.20(dd,J=13.2,7.9Hz,3H),2.47(td,J =9.8,4.8Hz,1H),1.96(dt,J=9.6,4.3Hz,1H),1.80(s,5H),1.42-1.29( m,2H),1.17-1.13(m,1H),0.93(d,J=1.7Hz,3H),0.90(d,J=2.7Hz,6H).

[0043] 13 C NMR(101MHz,Chloroform-d)δ165.17,158.17,137.61,136.47,135.91,128.47,127.92,11 9.87,84.96,67.87,49.40,48.23,44.87,36.63,27.99,27.31,19.86,19.71,18.91,13.66.

[0044] 1.4) Synthesis of 1-(3-methyl-2-enbutyl)-3-hydroxyisonicotinic acid dextran ester bromide (compound 4)

[0045] Referring to the synthesis method of compound 1, using isonicotinic acid dextran ester and 3-methyl-1-bromobutene as raw materials, iodomethane in step (2) of compound 1 was replaced with an equimolar amount of 3-methyl-1-bromobutene, and the rest were the same as step (2) of compound 1, to synthesize compound 4.

[0046]

[0047] Yellow solid; 1 H NMR (400MHz, Chloroform-d) δ9.15 (s, 1H), 9.03 (d, J = 6.2 Hz, 1H), 8.20 (d, J = 6. 1Hz,1H),5.50(s,3H),5.17-5.12(m,1H),2.43(tt,J=9.1,4.6Hz,1H),1.99-1. 93(m,1H),1.90(s,3H),1.87(s,1H),1.83(s,3H),1.76(d,J=9.3Hz,1H),1.42- 1.24(m,2H),1.12(dd,J=14.1,3.8Hz,1H),0.91(s,3H),0.87(d,J=5.1Hz,6H).

[0048] 13 C NMR(101MHz,Chloroform-d)δ165.09,157.88,146.45,135.55,128.31,127.97,115.25,84.71 ,60.21,49.36,48.20,44.85,36.62,34.59,27.98,27.29,26.07,19.70,19.13,18.89,13.65.

[0049] 1.5) Synthesis of 1-(2-encyclohexyl)-3-hydroxyisonicotinic acid dextran ester bromide (compound 5)

[0050] Referring to the synthesis method of compound 1, using isonicotinic acid dextran ester and 1-bromo-2-encyclohexene as raw materials, iodomethane in step (2) of compound 1 was replaced with an equimolar amount of 1-bromo-2-encyclohexene, and the rest were the same as step (2) of compound 1, to synthesize compound 5.

[0051]

[0052] Yellow solid; ¹H NMR (400 MHz, Chloroform-d) δ 9.32–9.12 (m, 2H), 8.27 (s, 1H), 6.46 (dt, J = 7.6, 3.7 Hz, 1H), 6.03–5.80 (m, 2H), 5.16 (d, J = 9.7 Hz, 1H), 2.46 (m, J = 19.0, 9.6, 5.4 Hz, 2H), 2.33 -2.14(m,2H),1.97(m,J=23.4,11.8,6.4Hz,2H),1.87-1.74(m,3H),1.66(dd,J=15.0, 7.6Hz,1H),1.42-1.25(m,2H),1.17-1.11(m,1H),0.92(s,3H),0.88(d,J=3.2Hz,6H).

[0053] 13 C NMR(101MHz,Chloroform-d)δ165.01,157.87,139.22,135.04,134.73,128.94,128.38,121.48,8 4.82,67.96,49.38,48.22,44.86,36.67,32.40,28.02,27.33,24.55,19.71,18.91,18.64,13.68.

[0054] 1.6) Synthesis of 1-benzyl-3-hydroxyisonicotinic acid dextran ester bromide (compound 6)

[0055]

[0056] Referring to the synthesis method of compound 1, using isonicotinic acid dextran ester and benzyl bromide as raw materials, iodomethane in step (2) of compound 1 was replaced with an equimolar amount of benzyl bromide, and the rest were the same as step (2) of compound 1, to synthesize compound 6.

[0057] Yellow solid; 1 H NMR (400MHz, Chloroform-d) δ9.15 (s, 1H), 9.03 (d, J = 6.2 Hz, 1H), 8.20 (d, J = 6. 1Hz,1H),5.50(s,3H),5.17-5.12(m,1H),2.43(tt,J=9.1,4.6Hz,1H),1.99-1. 93(m,1H),1.90(s,3H),1.87(s,1H),1.83(s,3H),1.76(d,J=9.3Hz,1H),1.42- 1.24(m,2H),1.12(dd,J=14.1,3.8Hz,1H),0.91(s,3H),0.87(d,J=5.1Hz,6H).

[0058] 13 C NMR(101MHz,Chloroform-d)δ165.09,157.88,146.45,135.55,128.31,127.97,115.25,84.71 ,60.21,49.36,48.20,44.85,36.62,34.59,27.98,27.29,26.07,19.70,19.13,18.89,13.65.

[0059] 1.7) Synthesis of 1-(4-methylbenzyl)-3-hydroxyisonicotinic acid dextran ester bromide (compound 7)

[0060] Referring to the synthesis method of compound 1, isonicotinic acid dextranol ester and 4-methylbenzyl bromide were used as raw materials. Iodomethane in step (2) of compound 1 was replaced with an equimolar amount of 4-methylbenzyl bromide. All other steps were the same as in step (2) of compound 1, and compound 7 was synthesized.

[0061]

[0062] Yellow solid; 1HNMR(400MHz,DMSO-d6)δ8.67(s,2H),8.19(d,J=6.1Hz,1H),7.41(d,J=7.7H z,2H),7.23(d,J=7.8Hz,2H),5.77(s,2H),5.03(d,J=9.8Hz,1H),2.35(d,J= 11.0Hz,1H),2.27(s,3H),1.91(d,J=11.7Hz,1H),1.68(q,J=8.2,5.2Hz,2H) ,1.24(dd,J=35.3,11.3Hz,2H),1.12-1.06(m,1H),0.87(s,3H),0.83(s,6H).

[0063] 13 C NMR(101MHz,DMSO-d6)δ163.63,155.90,139.66,135.76,135.67,132.66,131.41,130.29,129.6 7,128.71,82.51,63.86,49.34,48.15,44.74,36.52,28.00,27.27,21.30,20.01,19.15,13.89.

[0064] 1.8) Synthesis of 1-(4-fluorobenzyl)-3-hydroxyisonicotinic acid dextran ester bromide (compound 8)

[0065] Referring to the synthesis method of compound 1, using isonicotinic acid dextran ester and 4-fluorobenzyl bromide as raw materials, iodomethane in step (2) of compound 1 was replaced with an equimolar amount of 4-fluorobenzyl bromide, and the rest were the same as step (2) of compound 1, to synthesize compound 8.

[0066]

[0067] Yellow solid; 1 H NMR(400MHz,Chloroform-d)δ9.39(s,1H),9.31(d,J=6.1Hz,1H),8.18(t,J=4.3H z,1H),7.83-7.73(m,2H),7.03(td,J=8.5,3.0Hz,2H),6.32(d,J=2.6Hz,2H),5.1 6-5.09(m,1H),2.43(m,J=9.7,3.9Hz,1H),1.89(m,J=13.2,9.2,4.6Hz,1H),1.82 -1.68(m,2H),1.44-1.21(m,2H),1.09(dt,J=14.2,3.2Hz,1H),0.93-0.82(m,9H).

[0068] 13 C NMR(101MHz,Chloroform-d)δ165.39,164.94,162.45,158.14,136.58,135.87,132.28,128.58,127.83 ,127.39,116.88,116.66,84.98,64.00,49.37,48.22,44.82,36.57,27.95,27.27,19.68,18.87,13.62.

[0069] 1.9) Synthesis of 1-(4-trifluoromethylbenzyl)-3-hydroxyisonicotinic acid dextran ester bromide (compound 9)

[0070] Following the synthesis method of compound 1, isonicotinic acid dextranol ester and 4-trifluoromethylbenzyl bromide were used as raw materials. In step (2) of compound 1, iodomethane was replaced with an equimolar amount of 4-trifluoromethylbenzyl bromide. All other steps were the same as in step (2) of compound 1, and compound 9 was synthesized.

[0071]

[0072] Yellow solid; 1 HNMR(400MHz,Chloroform-d)δ9.18(s,1H),8.68(d,J=6.2Hz,1H),8.05(d,J=6.3Hz,1H) ,7.80(d,J=7.9Hz,2H),7.61(d,J=8.1Hz,2H),6.24(s,2H),5.10(dd,J=9.9,2.9Hz,1H), 2.41(ddt,J=14.0,8.6,4.0Hz,1H),1.93(m,J=13.3,9.1,4.0Hz,1H),1.82-1.70(m,2H), 1.36-1.22(m,2H),1.08(dd,J=13.9,3.4Hz,1H),0.90(s,3H),0.88(s,3H),0.85(s,3H).

[0073] 13 C NMR(101MHz,Chloroform-d)δ164.60,160.47,138.07,136.60,132.33,132.01,131.89,130.07,129.47,128.20,126 .64,126.60,126.56,124.97,122.26,83.75,63.82,49.28,48.13,44.87,36.70,27.99,27.30,19.69,18.87,13.63.

[0074] 1.10) Synthesis of 1-(4-methoxybenzyl)-3-hydroxyisonicotinic acid dextran ester bromide (compound 10)

[0075] Following the synthesis method of compound 1, isonicotinic acid dextranol ester and 4-methoxybenzyl bromide were used as raw materials. In step (2) of compound 1, iodomethane was replaced with an equimolar amount of 4-methoxybenzyl bromide. All other steps were the same as in step (2) of compound 1, and compound 10 was synthesized.

[0076]

[0077] Yellow solid; 1 H NMR(400MHz,Chloroform-d)δ9.46-9.10(m,2H),8.22-8.11(m,1H),7.63(d,J=7.9Hz, 2H),6.85(d,J=7.6Hz,2H),6.18(s,2H),5.16-5.10(m,1H),3.75(s,3H),2.43(ddt,J=1 3.9,8.8,4.0Hz,1H),1.92(m,J=13.3,9.1,4.0Hz,1H),1.76(dd,J=13.5,4.3Hz,2H),1 .43-1.23(m,2H),1.10(dd,J=14.0,3.5Hz,1H),0.91(s,3H),0.88(s,3H),0.86(s,3H).

[0078] 13 C NMR(101MHz,Chloroform-d)δ165.38,161.03,158.00,136.25,135.78,131.75,127.89,127.48,124.3 1,115.09,84.86,64.99,55.50,49.37,48.21,44.84,36.59,31.57,27.97,27.29,19.69,18.88,13.64.

[0079] 1.11) Synthesis of 1-(4-methylcarbamate benzyl)-3-hydroxyisonicotinic acid dextranol ester bromide (compound 11)

[0080] Referring to the synthesis method of compound 1, using isonicotinic acid dextranol ester and methyl 4-carboxylic acid benzyl bromide as raw materials, iodomethane in step (2) of compound 1 was replaced with an equimolar amount of methyl 4-carboxylic acid benzyl bromide, and the rest were the same as step (2) of compound 1, to synthesize compound 11.

[0081]

[0082] Yellow solid; 1 H NMR (400MHz, DMSO-d6) δ8.70(d,J=11.1Hz,2H),8.22(d,J=6.2Hz,1H),7.98(d,J= 7.9Hz,2H),7.62(d,J=8.1Hz,2H),5.93(s,2H),5.04(d,J=9.8Hz,1H),3.82(s,3H) ,2.35(td,J=10.3,9.8,5.2Hz,1H),1.93(m,J=12.7,9.3,4.2Hz,1H),1.74-1.64( m,2H),1.30-1.17(m,2H),1.09(dd,J=13.8,3.5Hz,1H),0.88(s,3H),0.83(s,6H).

[0083] 13 C NMR(101MHz,DMSO-d6)δ166.25,163.63,156.08,139.46,136.19,136.11,132.89,130.87,130.40,1 29.69,128.86,82.54,63.36,52.91,49.35,48.15,44.74,36.52,28.01,27.27,20.01,19.15,13.90.

[0084] 1.12) Synthesis of 1-(4-carboxybenzyl)-3-hydroxyisonicotinic acid dextran ester bromide (compound 12)

[0085] Referring to the synthesis method of compound 1, isonicotinic acid dextranol ester and 4-carboxylic acid benzyl bromide were used as raw materials. In step (2) of compound 1, iodomethane was replaced with an equimolar amount of 4-carboxylic acid benzyl bromide. All other steps were the same as in step (2) of compound 1, and compound 11 was synthesized.

[0086]

[0087] Yellow solid; 1H NMR (400MHz, DMSO-d6) δ8.77-8.66(m,2H),8.22(d,J=6.0Hz,1H),7.96(d,J=8.1H z,2H),7.61(d,J=8.2Hz,2H),5.95(s,2H),5.04(dt,J=9.8,2.9Hz,1H),2.34(ddt ,J=13.8,9.9,3.9Hz,1H),1.93(m,J=12.9,9.4,4.1Hz,1H),1.68(m,J=4.5,4.0Hz ,2H),1.31-1.16(m,2H),1.09(dd,J=13.8,3.4Hz,1H),0.87(s,3H),0.82(s,6H).

[0088] 13 C NMR(101MHz,DMSO-d6)δ167.29,163.60,156.00,138.96,136.18,136.09,132.87,132.11,130.53,1 29.59,128.84,82.54,63.40,55.47,49.34,48.14,44.73,36.51,28.00,27.27,20.00,19.14,13.90.

[0089] 1.13) Synthesis of 1-(3,3-dimethyl-2-butanone)-3-hydroxyisonicotinic acid dextran ester bromide (compound 13)

[0090] Referring to the synthesis method of compound 1, isonicotinic acid dextran ester and 1-bromo-3,3-dimethyl-2-butanone were used as raw materials. In step (2) of compound 1, iodomethane was replaced with an equimolar amount of 1-bromo-3,3-dimethyl-2-butanone. All other steps were the same as in step (2) of compound 1, and compound 13 was synthesized.

[0091]

[0092] Yellow solid; 1H NMR (400MHz, DMSO-d6) δ 8.52 (s, 1H), 8.43 (d, J = 6.3Hz, 1H), 8.26 (d, J = 6.1Hz, 1H), 6.04 (s, 2H), 5.07 (d, J = 9.8Hz, 1H), 2.37 (td, J = 10.1, 5.1Hz, 1H), 1.96 (m, J = 12.9, 9.6, 4.4Hz, 1H), 1.75–1.67 (m, 2H), 1.35–1.23 (m, 2H), 1.19 (s, 9H), 1.12 (d, J = 4.1Hz, 1H), 0.89 (s, 3H), 0.85 (d, J = 3.9Hz, 6H).

[0093] 13 C NMR(101MHz,DMSO-d6)δ206.94,163.71,155.28,137.39,137.28,133.06,128.07,82.64,6 6.00,49.37,48.17,44.76,43.59,36.51,31.55,28.01,27.31,26.19,20.03,19.17,13.93.

[0094] 1.14) Synthesis of 1-(3-phenyl-2-keto)-3-hydroxyisonicotinic acid dextran ester bromide (compound 14)

[0095] Referring to the synthesis method of compound 1, isonicotinic acid dextran ester and 1-bromo-3-phenyl-2-one were used as raw materials. In step (2) of compound 1, iodomethane was replaced with an equimolar amount of 1-bromo-3-phenyl-2-one. All other steps were the same as in step (2) of compound 1 to synthesize compound 14.

[0096]

[0097] Yellow solid; 1H NMR (400MHz, DMSO-d6) δ8.66(s,1H),8.55(d,J=6.3Hz,1H),8.32(d,J=6.0Hz,1H),8.02(d,J= 7.8Hz,2H),7.76(t,J=7.4Hz,1H),7.63(t,J=7.7Hz,2H),6.46(s,2H),5.09(d,J=9.7Hz,1H), 2.37(dd,J=9.7,4.4Hz,1H),1.99(m,J=12.7,9.2,4.1Hz,1H),1.70(dd,J=8.8,4.5Hz,2H),1. 28(dt,J=26.8,10.9Hz,2H),1.16(dd,J=13.9,3.4Hz,1H),0.90(s,3H),0.86(d,J=6.6Hz,6H).

[0098] 13 C NMR(101MHz,DMSO-d6)δ190.93,163.69,155.38,137.71,137.52,135.28,134.08,133.25,129.68,1 28.85,128.17,82.64,67.28,49.39,48.18,44.77,36.53,31.59,28.02,27.33,20.04,19.18,13.94.

[0099] 1.15) Synthesis of 1-(2-acetamido)-3-hydroxyisonicotinic acid dextran ester bromide (compound 15)

[0100] Following the synthesis method of compound 1, isonicotinic acid dextran ester and 1-bromo-2-acetamide were used as raw materials. In step (2) of compound 1, iodomethane was replaced with an equimolar amount of 1-bromo-2-acetamide. All other steps were the same as in step (2) of compound 1, and compound 15 was synthesized.

[0101]

[0102] Yellow solid; 1H NMR (400MHz, DMSO-d6) δ8.57(s,1H),8.52(d,J=6.3Hz,1H),8.21(d,J=6.2Hz,1H ),7.99(s,1H),7.69(s,1H),5.38(s,2H),5.06(dt,J=9.7,2.8Hz,1H),2.38(dt, J=14.0,4.2Hz,1H),1.95(m,J=12.8,9.4,4.2Hz,1H),1.70(q,J=4.2Hz,2H),1.3 3-1.22(m,2H),1.13(dd,J=13.9,3.5Hz,1H),0.89(s,3H),0.85(d,J=3.1Hz,6H).

[0103] 13 C NMR(101MHz,DMSO-d6)δ166.39,163.71,155.00,137.61,137.56,133.01,127.72,8 2.59,62.43,49.37,48.17,44.75,36.51,31.51,27.99,27.30,20.03,19.17,13.92.

[0104] 1.16) Synthesis of 1-(2-ethyl acetate)-3-hydroxyisonicotinic acid dextran ester bromide (compound 16)

[0105] Following the synthesis method of compound 1, using isonicotinic acid dextran ester and ethyl 1-bromo-2-acetate as raw materials, iodomethane in step (2) of compound 1 was replaced with an equimolar amount of ethyl 1-bromo-2-acetate, and the rest were the same as step (2) of compound 1, to synthesize compound 16.

[0106]

[0107] Yellow solid; ¹H NMR (400 MHz, Chloroform-d) δ 9.29 (s, ¹H), 9.12 (d, J = 6.3 Hz, ¹H), 8.22 (d, J = 6.0 Hz, ¹H), 6.24 (s, 2H), 5.16 (d, J = 9.8 Hz, 1H), 3.80 (s, 3H), 2.45 (ddt, J = 14.2, 9.2, 4.3 Hz, 1H), 1.94 (m, J = 13.5, 9.1, 3.8 Hz, 1H), 1.77 (dd, J = 14.5, 4.4 Hz, 2H), 1.43–1.25 (m, 2H), 1.14 (dd, J = 13.9, 3.4 Hz, 1H), 0.92 (s, 3H), 0.88 (s, 6H).

[0108] 13 C NMR(101MHz,Chloroform-d)δ166.02,165.32,157.74,138.62,137.59,128.30,127.27 ,84.92,61.63,53.98,49.40,48.23,44.85,36.57,27.96,27.29,19.70,18.89,13.65.

[0109] 1.17) Synthesis of 1-(2-Acetylphenyl)-3-hydroxyisonicotinic acid dextranol ester bromide (compound 17)

[0110] Referring to the synthesis method of compound 1, using isonicotinic acid dextran ester and 1-bromo-2-acetic acid phenyl ester as raw materials, iodomethane in step (2) of compound 1 was replaced with an equimolar amount of 1-bromo-2-acetic acid phenyl ester, and the rest were the same as step (2) of compound 1, to synthesize compound 17.

[0111]

[0112] Yellow solid; 1 HNMR(400MHz,Chloroform-d)δ9.40(s,1H),9.25(d,J=6.3Hz,1H),8.13(d,J=6.3 Hz,1H),7.31-7.16(m,5H),6.62(s,2H),5.13(dt,J=9.9,2.9Hz,1H),2.46(m,J=1 3.7,8.8,4.4Hz,1H),1.91(m,J=13.3,9.1,4.1Hz,1H),1.84-1.74(m,2H),1.43-1 .22(m,2H),1.12(dd,J=14.1,3.4Hz,1H),0.93(s,3H),0.90(s,3H),0.88(s,3H).

[0113] 13 C NMR(101MHz,Chloroform-d)δ165.44,164.69,157.81,150.03,138.67,137.75,129.66,129.53,127.68,127 .20,126.74,121.66,115.54,85.13,61.85,49.41,48.24,44.83,36.55,27.98,27.29,19.70,18.89,13.67.

[0114] Example 2

[0115] Effect of the target compound on the in vitro proliferation of RAW264.7 macrophages

[0116] RAW264.7 macrophages were digested, counted, and prepared into 7×10⁻⁶ cells. 4 RAW264.7 cell suspension (cells / mL) was added to each well of a 96-well cell culture plate at 100 μL. The plates were then incubated at 37°C with 5% CO2 for 24 h. The old culture medium was discarded, and 100 μL of the corresponding drug-containing culture medium (diluted to 6 μmol / L with the culture medium) was added to each well of the experimental group. A negative control group was also set up (100 μL of drug-free culture medium was added to each well). The plates were incubated at 37°C with 5% CO2 for 24 h. 20 μL of CCK-8 was added to each well, and the plates were incubated for another 2-3 h. The plates were then gently mixed for 10 min on a shaker to remove air bubbles. The OD value of each well was read using a microplate reader at λ = 450 nm, and the inhibition rate was calculated. The results are shown in Table 1.

[0117] Inhibition rate (%) = (OD value of negative control group - OD value of experimental group) / OD value of negative control group × 100%

[0118] Table 1. Inhibitory effect of the compound on the growth of RAW264.7 macrophages (6 μmol / L)

[0119] compound Inhibition rate compound Inhibition rate compound Inhibition rate Compound 1 C Compound 7 B Compound 13 C Compound 2 B Compound 8 B Compound 14 C Compound 3 B Compound 9 B Compound 15 C Compound 4 B Compound 10 B Compound 16 C Compound 5 B Compound 11 C Compound 17 C Compound 6 B Compound 12 C Compound 1a C

[0120] Note: A: 20%-11%, B: 10%-5%, C: 5%-0%.

[0121] As can be seen from Table 1, at a concentration of 6 μmol / L, the cytotoxicity of the compounds of this invention is less than 10%, indicating that they have good safety.

[0122] Example 3

[0123] Effects of the target compound on lipopolysaccharide-induced inflammatory factors in RAW264.7 macrophages

[0124] RAW264.7 cells were digested, counted, and prepared into 7×10⁻⁶ cells. 4RAW264.7 cell suspension was added to each well of a 96-well cell culture plate at a concentration of 100 μL / mL. The plates were then incubated at 37°C with 5% CO2 for 24 h. The old culture medium was discarded, and 100 μL of the corresponding drug-containing culture medium (diluted to 1 μmol / L) was added to each well of the experimental group. A negative control group (100 μL of drug-free culture medium per well) and a model group (100 μL of drug-free culture medium per well) were also set up. The plates were incubated at 37°C with 5% CO2 for 2 h. Lipopolysaccharide (LPS, final concentration 100 μg / L) was added to both the experimental and model groups and incubated for another 24 h. The supernatant was collected, and the levels of IL-1β and TNF-α were detected according to the ELISA kit instructions, as shown in Table 2.

[0125] Table 2. Effects of compounds on lipopolysaccharide-induced inflammatory factors in RAW264.7 macrophages (1 μmol / L)

[0126] TNF-α IL-1β TNF-α IL-1β TNF-α IL-1β negative control group 52.5 22.7 Compound 6 C E Compound 13 B D Model group 208.4 97.5 Compound 7 C E Compound 14 B D Compound 1 B D Compound 8 C E Compound 15 B D Compound 2 C E Compound 9 C E Compound 16 B D Compound 3 C E Compound 10 C E Compound 17 B D Compound 4 C E Compound 11 B D Compound 1a A C Compound 5 C E Compound 12 B D

[0127] Note: A: 200-151, B: 150-101, C: 100-76, D: 75-51, E: 50-25.

[0128] As can be seen from Table 2, the compounds of the present invention have a significant inhibitory effect on the increase of inflammatory factors in RAW264.7 macrophages induced by lipopolysaccharide.

[0129] Example 4

[0130] The compound (intraperitoneal injection) inhibited the carrageenan-induced paw edema in rats.

[0131] Male SD rats (SPF grade, 6-8 weeks old, weighing 250-280g) were randomly divided into groups of 6. The normal control group and the model group were intravenously injected with an equal volume of solvent (5% DMSO). Each drug group received an intraperitoneal injection of 5 mg / kg of the respective compound (compound concentration 0.5 mg / mL). 0.5 h after drug administration, the normal control group received a subcutaneous injection of 100 μL of physiological saline into the hind limb toes, while the other groups received an injection of 100 μL of 1% carrageenan. The paw volume was measured at 0 h, 1 h, and 2 h after carrageenan injection, and the paw edema rate (%) was calculated.

[0132] Foot edema rate = (post-inflammatory plantar volume - pre-inflammatory plantar volume) / pre-inflammatory plantar volume × 100%.

[0133] The results are as follows Figure 1 As shown, the compound of the present invention has a significant inhibitory effect on carrageenan-induced paw edema in rats.

[0134] Example 5

[0135] Compound 6 single intravenous injection drug metabolism study in rats

[0136] Animals: SD rats, male, 3 rats per group.

[0137] Solvent: 5% DMSO + 20% PEG400 + 10% Solutol + 65% physiological saline.

[0138] Blood collection time: 0.0833h, 0.25h, 0.5h, 1h, 2h, 4h, 6h, 8h, 24h.

[0139] The main pharmacokinetic parameters of compound 6 after a single intravenous (IV) injection of 2 mg / kg in rats were: C max It was 24.3 ng / mL, T max It is 0.0833h, T 1 / 2 The AUC is 0.166h. 0-T It was 7.97 hr·ng / mL, AUC 0-∞ It was 8.43 hr·ng / mL, V z The concentration was 62316 mL / kg, the CL was 259712 mL / hr / kg, and the MRT was... 0-t For 0.163h, MRT 0-∞ It is 0.208h.

[0140] Preliminary drug metabolism studies showed that compound 6 was rapidly metabolized in animals. The other 16 target compounds also exhibited similar characteristics to compound 6 in animals, with short half-lives.

[0141] Example 6

[0142] Compound 6's inhibitory effect on acute ocular inflammation induced by 30% croton oil in rabbits

[0143] Animal: Domestic rabbit.

[0144] Solvent: 0.1% poloxamer + 0.1% nonyl alcohol ether 4 + 1.8% mannitol + 2.0% glycerol + 95.8% physiological saline.

[0145] Three rabbits were included in each group: a compound 6 eye drop group (2.0 mg / mL, prepared using a solvent), a positive control group (0.5% cortisone acetate eye drops), and a negative control group (0.1% poloxamer + 0.1% nonoxynol ether 4 + 1.8% mannitol + 2.0% glycerol + 95.8% physiological saline). 50 μL of 30% croton oil was accurately instilled into the rabbits' eyes using a micropipette. After instillation of croton oil, instillation continued at 0.5 h intervals for a total of four instillations. After two h, instillation was reduced to once every 1 h for a total of four instillations, with 50 μL instilled each time. Changes in conjunctival inflammation were scored according to the conjunctival inflammation scoring criteria at 4 h and 8 h after croton oil instillation.

[0146] Table 3. Scoring criteria for conjunctivitis

[0147] Eye symptoms Score 1. Conjunctival congestion (1) Normal blood vessels 0 (2) The blood vessels are congested and appear bright red. 1 (3) The blood vessels are congested and appear dark red, making them difficult to distinguish. 2 (4) Diffuse congestion appears purplish-red. 3 2. Conjunctival edema (1) No edema 0 (2) Mild edema 1 (3) Obvious edema accompanied by partial ectropion 2 (4) Edema to the point that the eyelids are almost half closed 3 3. Secretions (1) No discharge 0 (2) Small amount of secretions 1 (3) Secretions make the eyelids and eyelashes moist or sticky. 2 (4) Secretions make the entire eye area moist or sticky. 3

[0148] Table 4. Effect of compound 6 on croton oil-induced conjunctivitis in rabbits (n = 3, X ± SD)

[0149] Inflammation score 4h Inflammation score 8h negative control group 8.55±0.36 8.32±0.33 Positive control group 4.89±0.69 3.15±0.48 Compound 6 eye drops group 5.06±0.47 3.32±0.41

[0150] The experimental results are shown in Table 4. It can be seen that the compound of the present invention has a significant inhibitory effect on croton oil-induced conjunctivitis in rabbits.

Claims

1. Quaternary ammonium salts of 3-hydroxyisonicotinic acid dextran ester with the structure shown in Formula I: in, R is selected from saturated or unsaturated C1-C6 hydrocarbon groups, substituted or unsubstituted benzyl groups, The substituents of the benzyl group are selected from halogens, C1-C3 alkyl groups, C1-C3 alkoxy groups, halogen-substituted C1-C3 alkyl groups, etc. R1 is selected from NH2, C1-C4 alkyl, C1-C4 alkoxy, phenyl, phenoxy; R2 is selected from H, C1-C3 alkyl; X is selected from bromine, iodine.

2. The quaternary ammonium salt compound of 3-hydroxyisonicotinic acid dextran ester according to claim 1, characterized in that: R is selected from substituted or unsubstituted benzyl groups. C1-C3 alkyl groups The substituents of benzyl are selected from fluorine, methyl, methoxy, trifluoromethane, The substituents can be located at the ortho, meta, or para position of existing substituents on the benzene ring; R1 is selected from NH2, C1-C4 alkyl, C1-C4 alkoxy, phenyl, and phenoxy; R2 is selected from H and methyl; R3, R4, and R5 are each independently selected from H and methyl; X = bromine or iodine.

3. The quaternary ammonium salt compound of 3-hydroxyisonicotinic acid dextranol ester according to claim 2, characterized in that: R is selected from substituted or unsubstituted benzyl groups. The substituents of benzyl are selected from fluorine, methyl, methoxy, and trifluoromethane, and the substituents are located at the para position of the existing substituents on the benzene ring; R3 = R4 = R5 = H or R3 = methyl, R4 = R5 = H or R3 = H, R4 = R5 = methyl; X = bromine or iodine.

4. Quaternary ammonium salts of 3-hydroxyisonicotinic acid dextran ester with the structure shown below:

5. Use of the quaternary ammonium salt compound of 3-hydroxyisonicotinic acid dextran ester according to any one of claims 1-4 in the preparation of a medicament for treating inflammation-related diseases.

6. The use according to claim 5, characterized in that: The inflammation-related diseases mentioned are rheumatoid arthritis, eye inflammation, and respiratory tract inflammation.

7. A pharmaceutical composition for treating inflammation-related diseases, characterized in that: The pharmaceutical composition comprises, as the active ingredient, a quaternary ammonium salt of 3-hydroxyisonicotinic acid dextran ester as described in any one of claims 1-4, and formulated into a pharmaceutically acceptable dosage form with pharmaceutically acceptable excipients.

8. The pharmaceutical composition according to claim 7, characterized in that: The dosage forms mentioned are solutions, sprays, patches, and gels.

9. Use of the pharmaceutical composition of claim 7 in the preparation of a medicament for treating inflammation-related diseases.

10. The use according to claim 9, characterized in that: The inflammation-related diseases mentioned are rheumatoid arthritis, eye inflammation, and respiratory tract inflammation.

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