Derivative of artemisinin and application of derivative in treatment of cerebral apoplexy

Abstract

The invention discloses an artemisinin derivative and application of the artemisinin derivative in treatment of cerebral apoplexy. The structural formula of the artemisinin derivative is as shown in formula I in the specification. Pharmacodynamic experiments show that the artemisinin derivative provided by the invention can significantly improve the falling latency, behavioral score, cerebral infarction area percentage and cerebral blood flow improvement percentage of a model animal, is superior to other artemisinin derivatives at present, and is an excellent candidate drug for treating cerebral apoplexy. # imgabs0 #

Description

Technical Field

[0001] The invention belongs to the field of medicine, and specifically relates to an artemisinin derivative and an application thereof in treating cerebral stroke. Background Art

[0002] Acute ischemic stroke (AIS) is a cerebrovascular disease caused by vascular blockage. Although clinical treatment measures have been significantly improved, it is still the most common cause of disability in adults worldwide. After AIS occurs, it quickly causes brain tissue death in the ischemic core area, which in turn leads to delayed death of neurons in the ischemic peri-area (also known as the penumbra). The penumbra has reversible damage, and saving and protecting the ischemic penumbra through reasonable therapeutic intervention has become a breakthrough in the prevention and treatment of this disease.

[0003] Studies have revealed that artemisinin (ART) has multiple biological effects such as anti-apoptosis and cardiovascular protection. Its mechanism of action involves activating the ERK1 / 2 / CREB / bcl2 signaling pathway, thereby alleviating IS-induced apoptosis. In addition, the results also showed that ART can significantly reduce the molecular markers of apoptosis, oxidative stress, and neuroinflammation levels.

[0004] However, due to various factors, no artemisinin-based drugs have been marketed for the treatment of damage caused by acute ischemic stroke, and more effective derivatives of artemisinin for the treatment of stroke are still being explored. Summary of the invention

[0005] In view of the defects of the prior art, the present invention develops a new class of artemisinin derivatives for treating stroke.

[0006] The novel artemisinin derivative for treating cerebral stroke provided by the present invention has a structural formula as shown in Formula I:

[0007]

[0008] The pharmaceutically acceptable salts, esters, solvates of the compound of formula (I) and the diastereomers of the compound of formula (I) and their pharmaceutically acceptable salts, esters, solvates all fall within the protection scope of the present invention.

[0009] X in the above formula (I) - Can represent anions generated by ionization of inorganic or organic acids.

[0010] Furthermore, the inorganic acid or organic acid includes phosphoric acid, sulfuric acid, nitric acid, hydrochloric acid, iodic acid, bromic acid, methanesulfonic acid, maleic acid, fumaric acid, succinic acid, tartaric acid and the like.

[0011] In the above formula (I), n is an integer of 0-5, specifically 0, 1, 2, 3, 4 or 5.

[0012] The pharmaceutically acceptable salt of the compound represented by formula (I) in the present invention refers to a salt that is suitable for contact with the tissues of humans and lower animals without excessive toxicity, irritation, allergic reaction, etc., within the scope of reliable medical judgment, and is commensurate with a reasonable effect / risk ratio.

[0013] Preferably, the pharmaceutically acceptable salt of the compound represented by formula (I) is phosphate, sulfate, nitrate, hydrochloride, iodide, bromide, methanesulfonate, maleate, fumarate, succinate or tartrate.

[0014] The present invention also provides a method for preparing the compound represented by the above formula (I).

[0015] The preparation method of the compound represented by formula (I) provided by the present invention comprises the following steps: condensing the compound represented by formula (II) with an amino compound to generate the compound represented by formula (I), wherein the amino compound is trimethylamine or a salt thereof.

[0016]

[0017] In the formula (II), Y represents a halogen atom, including Cl, Br or I atom; n is any integer from 0 to 5, specifically 0, 1, 2, 3, 4, 5.

[0018] The preparation method of the compound represented by the above formula (II) is as follows: Compound c (dihydroartemisinin trimethylsilyl ether) is condensed with the compound represented by formula (III) to obtain the compound.

[0019]

[0020] The definition of Y in the formula (III) is the same as that of the formula (II); n is any integer between 0 and 5.

[0021] The amino compound is preferably trimethylamine hydrochloride.

[0022] The present invention also provides the use of the compound represented by the above formula (I) or its pharmaceutically acceptable salt ester, solvate and the diastereomer of the compound of formula (I) and its pharmaceutically acceptable salt, ester, solvate in the preparation of drugs for preventing and / or treating cerebral stroke or cerebral ischemia-reperfusion injury.

[0023] Furthermore, the stroke is acute ischemic stroke.

[0024] Furthermore, the compound represented by formula (I) can reduce the cerebral infarction area and increase the cerebral blood flow in patients with ischemia-reperfusion injury.

[0025] The present invention also provides a medicine or pharmaceutical composition for preventing and / or treating cerebral stroke or cerebral ischemia-reperfusion injury, comprising a compound as shown in the above formula (I), a diastereomer, or a pharmaceutically acceptable salt ester or solvate thereof, and a pharmaceutically acceptable carrier.

[0026] The drug can be introduced into the body such as muscle, intradermal, subcutaneous, intravenous, mucosal tissue by injection, spraying, nasal drops, eye drops, penetration, absorption, physical or chemical mediation; or introduced into the body after being mixed or wrapped with other substances.

[0027] Preferably, the pharmaceutically acceptable carrier comprises at least one of a diluent, an excipient, a filler, a binder, a wetting agent, a disintegrant, an absorption promoter, a surfactant, an adsorption carrier and a lubricant.

[0028] The above-mentioned medicine or pharmaceutical composition can be prepared into dosage forms such as solution, tablet, capsule or injection according to conventional methods known to those skilled in the art.

[0029] Beneficial effects: The present invention has found through pharmacodynamics experiments comparing a series of compounds that the compound represented by formula (I) is superior to dihydroartemisinin and other compounds and is an excellent candidate drug for treating stroke. BRIEF DESCRIPTION OF THE DRAWINGS

[0030] Figure 1 The synthetic flow chart of the compound represented by formula (I) is shown in FIG. DETAILED DESCRIPTION

[0031] The present invention is further described in detail below in conjunction with specific embodiments, and the examples provided are only for illustrating the present invention, rather than for limiting the scope of the present invention. The examples provided below can be used as a guide for further improvements by those of ordinary skill in the art, and do not constitute a limitation of the present invention in any way.

[0032] The experimental methods in the following examples, unless otherwise specified, are all conventional methods, and are performed according to the techniques or conditions described in the literature in the field or according to the product instructions. The materials, reagents, etc. used in the following examples, unless otherwise specified, can all be obtained from commercial channels.

[0033] Embodiment 1,

[0034] 1. (4-{[(1S,4S,5R,8S,9R,12R,13R)-1,5,9-trimethyl-11,14,15,16-tetraoxatetracyclo[10.3.1.0 4,13 .0 8,13 ]hexadec-10-yl]amino}butyl)trimethylammonium iodide (test drug 1)

[0035]

[0036] 1)(1S,4S,5R,8S,9R,12R,13R)-1,5,9-trimethyl-11,14,15,16-tetraoxatetracyclo[10.3.1.0 4,13 .0 8,13 ] Hexadecane-10-ol (Compound b)

[0037]

[0038] Artemisinin (10 g, 35.5 mmol) and methanol (100 mL) were added to a round-bottom flask, cooled to 0°C in an ice-water bath, and sodium borohydride (1.35 g, 35.5 mmol) was added in batches, and the reaction was kept at constant temperature for 2 h. After the reaction was completed, excess water was added to dilute the reaction solution, filtered, and the filter residue was vacuum dried to obtain a white solid powder (9.3 g, 92.3%). 1 HNMR(CDCl3 500MHz)δ5.63-4.73(m,2H),3.25(s,1H),2.97-0.98(m,21H).ESI-MS m / z:285.2[M+H] + .

[0039] 2) {[(1S,4S,5R,8S,9R,12R,13R)-1,5,9-trimethyl-11,14,15,16-tetraoxatetracyclo[10.3.1.0 4,13 .0 8,13 ]hexadec-10-yl]oxy}trimethylsilane (Compound c)

[0040]

[0041] (1S,4S,5R,8S,9R,12R,13R)-1,5,9-trimethyl-11,14,15,16-tetraoxatetracyclo[10.3.1.0 4,13 .0 8,13 Hexadecane-10-ol (9.0 g, 31.7 mmol) was dissolved in dichloromethane (100 mL), cooled to 0°C in an ice-water bath, and then trimethylsilyl chloride (5.18 g, 47.6 mmol) was added dropwise, and the mixture was reacted at a constant temperature for 3 h. After the reaction was completed, a saturated sodium bicarbonate aqueous solution was added to neutralize the reaction solution, the organic phase was concentrated, and a white solid powder (8.8 g, 78.3%) was obtained by column chromatography purification. 1 HNMR(CDCl3 500MHz)δ5.79(d,1H),5.44(s,1H),2.87-0.88(m,21H),0.43(s,9H).ESI-MS m / z:357.1[M+H] + .

[0042] 3)(4-chlorobutyl)[(1S,4S,5R,8S,9R,12R,13R)-1,5,9-trimethyl-11,14,15,16-tetraoxatetracyclo[10.3.1.0 4,13 .0 8,13 ]Hexadec-10-yl]amine (compound d, formula (II))

[0043]

[0044] {[(1S,4S,5R,8S,9R,12R,13R)-1,5,9-trimethyl-11,14,15,16-tetraoxatetracyclo[10.3.1.0 4,13 .0 8,13 ] Hexadecane-10-yl]oxy}trimethylsilane (8.0 g, 22.8 mmol) was dissolved in dichloromethane (100 mL), cooled to 0°C in an ice-water bath, and trimethylsilyl bromide (3.44 g, 22.8 mmol) was added dropwise, and the reaction was carried out at a constant temperature for 1 h. Then 4-chlorobutylamine hydrochloride (3.28 g, 22.8 mmol) was added, and the reaction was carried out at room temperature for 5 h. After the reaction was completed, a saturated sodium bicarbonate aqueous solution was added to neutralize the reaction solution, and the organic phase was concentrated and purified by column chromatography to obtain a light yellow oil (5.43 g, 63.8%). 1 H NMR (CDCl3 500MHz) δ5.47-4.81(m,2H),4.25-3.38(m,5H),2.77-0.98(m,15H).ESI-MS m / z:374.8[M+H] + .

[0045] 4)(4-{[(1S,4S,5R,8S,9R,12R,13R)-1,5,9-trimethyl-11,14,15,16-tetraoxatetracyclo[10.3.1.0 4,13 .0 8,13 ]hexadec-10-yl]amino}butyl)trimethylammonium iodide

[0046]

[0047] (4-chlorobutyl) [(1S,4S,5R,8S,9R,12R,13R)-1,5,9-trimethyl-11,14,15,16-tetraoxatetracyclo [10.3.1.0 4,13 .0 8,13]Hexadecane-10-yl]amine (5g, 13.4mmol), methanol (100mL), trimethylamine hydrochloride (12.9g, 134mmol), sodium iodide (2.0g, 13.4mmol), anhydrous potassium carbonate (1.85g, 13.4mmol) were added to a round-bottom flask and refluxed at 70°C for 24h. After the reaction was completed, the mixture was cooled and filtered, and the filtrate was concentrated under reduced pressure. After purification by column chromatography, a white foam powder (3.5g, 50.1%) was obtained. 1 H NMR (CDCl3 500MHz) δ5.43-4.74(m,2H),4.14-3.48(m,14H),2.81-0.95(m,25H).ESI-MS m / z:397.2[M+H] + .

[0048] Example 2. Preparation of other salts of the compound represented by formula (I)

[0049] 1. (4-{[(1S,4S,5R,8S,9R,12R,13R)-1,5,9-trimethyl-11,14,15,16-tetraoxatetracyclo[10.3.1.0 4,13 .0 8,13 ]hexadec-10-yl]amino}butyl)trimethylammonium phosphate

[0050]

[0051] (4-{[(1S,4S,5R,8S,9R,12R,13R)-1,5,9-trimethyl-11,14,15,16-tetraoxatetracyclo[10.3.1.0 4,13 .0 8,13 ]hexadec-10-yl]amino}butyl)trimethylammonium iodide (0.5 g, 0.95 mmol) was dissolved in water (50 mL) and desalted by AG MP-1M anion exchange resin. The eluate was neutralized with phosphoric acid and concentrated and recrystallized to obtain a light yellow (4-{[(1S,4S,5R,8S,9R,12R,13R)-1,5,9-trimethyl-11,14,15,16-tetraoxatetracyclo[10.3.1.0 4,13 .0 8,13 ]hexadec-10-yl]amino}butyl)trimethylammonium phosphate (0.42 g, 88.6%). 1 HNMR(CDCl3 500MHz)δ5.44-4.65(m,2H),4.18-3.45(m,14H),2.86-0.97(m,25H)..ESI-MS m / z:397.2[M+H] + .

[0052] 2. (4-{[(1S,4S,5R,8S,9R,12R,13R)-1,5,9-trimethyl-11,14,15,16-tetraoxatetracyclo[10.3.1.0 4,13 .0 8,13 ]hexadec-10-yl]amino}butyl)trimethylammonium sulfate

[0053]

[0054] Combining the ion exchange purification method in 1 above, (4-{[(1S,4S,5R,8S,9R,12R,13R)-1,5,9-trimethyl-11,14,15,16-tetraoxatetracyclo[10.3.1.0 4,13 .0 8,13 ]hexadec-10-yl]amino}butyl)trimethylammonium sulfate. 1 H NMR (CDCl3 500MHz) δ5.54-4.66(m,2H),4.24-3.56(m,14H),2.88-0.96(m,25H)..ESI-MS m / z:397.2[M+H] + .

[0055] 3. (4-{[(1S,4S,5R,8S,9R,12R,13R)-1,5,9-trimethyl-11,14,15,16-tetraoxatetracyclo[10.3.1.0 4,13 .0 8,13 ]hexadec-10-yl]amino}butyl)trimethylammonium nitrate

[0056]

[0057] Combining the ion exchange purification method in 1 above, (4-{[(1S,4S,5R,8S,9R,12R,13R)-1,5,9-trimethyl-11,14,15,16-tetraoxatetracyclo[10.3.1.0 4,13 .0 8,13 ]hexadec-10-yl]amino}butyl)trimethylammonium nitrate. 1 H NMR (CDCl3 500MHz) δ5.44-4.73(m,2H),4.24-3.53(m,14H),2.82-0.95(m,25H)..ESI-MS m / z:397.2[M+H] + .

[0058] 4. (4-{[(1S,4S,5R,8S,9R,12R,13R)-1,5,9-trimethyl-11,14,15,16-tetraoxatetracyclo[10.3.1.04,13 .0 8,13 ]hexadec-10-yl]amino}butyl)trimethylammonium hydrochloride (test drug 3)

[0059]

[0060] Combining the ion exchange purification method in 1 above, (4-{[(1S,4S,5R,8S,9R,12R,13R)-1,5,9-trimethyl-11,14,15,16-tetraoxatetracyclo[10.3.1.0 4,13 .0 8,13 ]hexadec-10-yl]amino}butyl)trimethylammonium hydrochloride. 1 H NMR (CDCl3 500MHz) δ5.38-4.67(m,2H),4.34-3.53(m,14H),2.96-0.97(m,25H)..ESI-MS m / z:397.2[M+H] + .

[0061] 5. (4-{[(1S,4S,5R,8S,9R,12R,13R)-1,5,9-trimethyl-11,14,15,16-tetraoxatetracyclo[10.3.1.0 4,13 .0 8,13 ]hexadec-10-yl]amino}butyl)trimethylammonium bromide

[0062]

[0063] Combining the ion exchange purification method in 1 above, (4-{[(1S,4S,5R,8S,9R,12R,13R)-1,5,9-trimethyl-11,14,15,16-tetraoxatetracyclo[10.3.1.0 4,13 .0 8,13 ]hexadec-10-yl]amino}butyl)trimethylammonium bromide. 1 H NMR (CDCl3 500MHz) δ5.36-4.68(m,2H),4.38-3.33(m,14H),2.88-0.97(m,25H)..ESI-MS m / z:397.2[M+H] + .

[0064] 6. (4-{[(1S,4S,5R,8S,9R,12R,13R)-1,5,9-trimethyl-11,14,15,16-tetraoxatetracyclo[10.3.1.0 4,13 .0 8,13 ]hexadec-10-yl]amino}butyl)trimethylammonium methanesulfonate

[0065]

[0066] Combining the ion exchange purification method in 1 above, (4-{[(1S,4S,5R,8S,9R,12R,13R)-1,5,9-trimethyl-11,14,15,16-tetraoxatetracyclo[10.3.1.0 4,13 .0 8,13 ]hexadec-10-yl]amino}butyl)trimethylammonium methanesulfonate. 1 H NMR (CDCl3 500MHz) δ5.38-4.66(m,2H),4.35-3.45(m,17H),2.93-0.97(m,25H)..ESI-MS m / z:397.2[M+H] + .

[0067] 7. (4-{[(1S,4S,5R,8S,9R,12R,13R)-1,5,9-trimethyl-11,14,15,16-tetraoxatetracyclo[10.3.1.0 4,13 .0 8,13 ]hexadec-10-yl]amino}butyl)trimethylammonium maleate

[0068]

[0069] Combining the ion exchange purification method of Example 4, (4-{[(1S,4S,5R,8S,9R,12R,13R)-1,5,9-trimethyl-11,14,15,16-tetraoxatetracyclo[10.3.1.0 4,13 .0 8,13 ]hexadec-10-yl]amino}butyl)trimethylammonium maleate. 1 H NMR(CDCl3 500MHz)δ13.0(s,1H),6.33(d,2H),5.38-4.66(m,2H),4.35-3.45(m,17H),2.93-0.97(m,25H)..ESI-MS m / z:397.2[M+H] + .

[0070] 8. (4-{[(1S,4S,5R,8S,9R,12R,13R)-1,5,9-trimethyl-11,14,15,16-tetraoxatetracyclo[10.3.1.0 4,13 .0 8,13 ]hexadec-10-yl]amino}butyl)trimethylammonium fumarate

[0071]

[0072] Combining the ion exchange purification method in 1 above, (4-{[(1S,4S,5R,8S,9R,12R,13R)-1,5,9-trimethyl-11,14,15,16-tetraoxatetracyclo[10.3.1.0 4,13 .0 8,13 ]hexadec-10-yl]amino}butyl)trimethylammonium fumarate. 1 H NMR (CDCl3 500MHz) δ13.0(s,1H),6.28(d,2H),5.36-4.62(m,2H),4.38-3.46(m,17H),2.84-0.96(m,25H)..ESI-MS m / z:397.2[M+H] + .

[0073] 9. (4-{[(1S,4S,5R,8S,9R,12R,13R)-1,5,9-trimethyl-11,14,15,16-tetraoxatetracyclo[10.3.1.0 4,13 .0 8,13 ]hexadec-10-yl]amino}butyl)trimethylammonium succinate

[0074]

[0075] Combining the ion exchange purification method in 1 above, (4-{[(1S,4S,5R,8S,9R,12R,13R)-1,5,9-trimethyl-11,14,15,16-tetraoxatetracyclo[10.3.1.0 4,13 .0 8,13 ]hexadec-10-yl]amino}butyl)trimethylammonium succinate. 1 H NMR (CDCl3 500MHz) δ13.0(s,1H),5.38-4.69(m,2H),4.33-3.47(m,17H),2.92-0.96(m,27H).ESI-MS m / z:397.2[M+H] + .

[0076] 10. (4-{[(1S,4S,5R,8S,9R,12R,13R)-1,5,9-trimethyl-11,14,15,16-tetraoxatetracyclo[10.3.1.0 4,13 .0 8,13 ]hexadec-10-yl]amino}butyl)trimethylammonium tartrate

[0077]

[0078] Combining the ion exchange purification method in 1 above, (4-{[(1S,4S,5R,8S,9R,12R,13R)-1,5,9-trimethyl-11,14,15,16-tetraoxatetracyclo[10.3.1.0 4,13 .0 8,13 ]hexadec-10-yl]amino}butyl)trimethylammonium tartrate. 1 H NMR (CDCl3 500MHz) δ13.0(s,1H),5.45-4.62(m,6H),4.36-3.46(m,17H),2.95-0.98(m,25H).ESI-MS m / z:397.2[M+H] + .

[0079] Comparative ratio,

[0080] 1. (4-{[(1S,4S,5R,8S,9R,12R,13R)-1,5,9-trimethyl-11,14,15,16-tetraoxatetracyclo[10.3.1.0 4,13 .0 8,13 ]hexadec-10-yl]amino}butyl)dimethylamine (test drug 4)

[0081]

[0082] Based on the synthesis method of Example 1, (4-chlorobutyl) [(1S, 4S, 5R, 8S, 9R, 12R, 13R) -1,5,9-trimethyl-11,14,15,16-tetraoxatetracyclo [10.3.1.0 4,13 .0 8,13 ] hexadecane-10-yl] amine and dimethylamine hydrochloride condensation to give (4-{[(1S,4S,5R,8S,9R,12R,13R)-1,5,9-trimethyl-11,14,15,16-tetraoxatetracyclo[10.3.1.0 4,13 .0 8,13 ]hexadec-10-yl]amino}butyl)dimethylamine 1 H NMR (CDCl3 500MHz) δ5.32-4.64(m,2H),4.08-3.26(m,11H),2.81-0.95(m,25H)..ESI-MS m / z:383.2[M+H] + .

[0083] 2. [4-(Methylamino)butyl][(1S,4S,5R,8S,9R,12R,13R)-1,5,9-trimethyl-11,14,15,16-tetraoxatetracyclo[10.3.1.0 4,13 .08,13 ]Hexadecane-10-yl]amine (test drug 2)

[0084]

[0085] Based on the synthesis method of Example 1, (4-chlorobutyl) [(1S, 4S, 5R, 8S, 9R, 12R, 13R) -1,5,9-trimethyl-11,14,15,16-tetraoxatetracyclo [10.3.1.0 4,13 .0 8,13 ] Hexadecane-10-yl]amine was condensed with methylamine aqueous solution to give [4-(methylamino)butyl][(1S,4S,5R,8S,9R,12R,13R)-1,5,9-trimethyl-11,14,15,16-tetraoxatetracyclo[10.3.1.0 4,13 .0 8,13 ]Hexadec-10-yl]amine 1 H NMR (CDCl3 500MHz) δ5.38-4.57(m,2H),4.11-3.17(m,8H),2.79-0.88(m,25H)..ESI-MS m / z:369.2[M+H] + .

[0086] 3. (4-{[(1R,3R,5R,6R,7S,10R,11S,12S)-1,6,10-trimethyl-2,4,13,14-tetraoxatetracyclo[9.3.2.0 3,12 .0 7,12 Synthesis of 5-(6-(6-hexadecyl)oxy)butyl)trimethylammonium iodide (Test drug 5)

[0087]

[0088] Add 1g of 3-dimethylamino-1-butanol to a 100ml single-necked flask, add 20ml of acetonitrile and stir to dissolve, add 3.64g of iodomethane dropwise, stir at room temperature for 2h, and monitor the reaction of the raw materials by TLC. Add 20ml of tert-methyl ether dropwise to the reaction solution, and a white solid precipitates in the solution. Filter by suction to obtain 2.0g of quaternary ammonium salt intermediate II.

[0089] 2.0g of quaternary ammonium salt intermediate II and 2.0g of dihydroartemisinin were added to a 100ml single-necked flask in sequence, and 40ml of dichloromethane was added for stirring and dispersion. A catalytic amount of boron trifluoride ether was added dropwise. After the addition was completed, the clear solution gradually turned dark red and stirred at room temperature for 2h. TLC monitored the completion of the reaction of the raw materials. The reaction solution was washed with saturated ammonium chloride and saturated sodium chloride in sequence, and the organic phase was dried under reduced pressure to obtain 2.5g of the crude product of the test drug 5; the crude product was purified by dichloromethane: methanol (1:0-30:1) column chromatography to obtain 300mg of the foamy solid test drug 5.

[0090] 1HNMR CDCl3δ:5.4(s,1H),5.0(m,1H),3.8(m,1H),3.6-3.5(m,4H),3.3(m,10H),2.2 (m,1H),2.0(m,1H),1.9-1.8(m,4H),1.6-1.3(m,11H),1.1(m,3H),0.8(m,3H). LC-MS: m / z=398.1.0(M+1).

[0091] 4. (Furan-2-ylmethyl)(2-{[(1S,3R,5S,6R,7S,10R,11S,12R)-1,6,10-trimethyl-2,4,13,14-tetraoxatetracyclo[9.3.2.0 3,12 .0 7,12 ]hexadec-5-yl]oxy}ethyl)amine (test drug 7)

[0092]

[0093] 1)(1S,3R,5S,6R,7S,10R,11S,12R)-5-[(2-bromoethyl)oxy]-1,6,10-trimethyl-2,4,13,14-tetraoxatetracyclo[9.3.2.0 3,12 .0 7,12 Hexadecane

[0094]

[0095] (1S,4S,5R,8S,9R,12R,13R)-1,5,9-trimethyl-11,14,15,16-tetraoxatetracyclo[10.3.1.0 4,13 .0 8,13 Hexadecane-10-ol (5.0 g, 17.6 mmol) and 2-bromoethanol (2.4 g, 19.4 mmol) were dissolved in tetrahydrofuran (100 mL), cooled to 0°C in an ice-water bath, and then boron trifluoride ether solution (4 mL) was added dropwise, and the reaction was kept at constant temperature for 1.5 h. After the reaction was completed, saturated sodium bicarbonate aqueous solution was added to neutralize the reaction solution, the organic phase was concentrated, and a white solid powder (5.9 g, 85.6%) was obtained by column chromatography purification. 1 H NMR (CDCl3 500MHz) δ5.89(d,1H),5.21(s,1H),4.13(m,2H),3.86(m,2H),2.93-0.89(m,21H).ESI-MS m / z:390.1[M] + ,392.1[M+2] + .

[0096] 2)(Furan-2-ylmethyl)(2-{[(1S,3R,5S,6R,7S,10R,11S,12R)-1,6,10-trimethyl-2,4,13,14-tetraoxatetracyclo[9.3.2.0 3,12 .0 7,12 ]hexadec-5-yl]oxy}ethyl)amine

[0097]

[0098] (1S,3R,5S,6R,7S,10R,11S,12R)-5-[(2-bromoethyl)oxy]-1,6,10-trimethyl-2,4,13,14-tetraoxatetracyclo[9.3.2.0 3,12 .0 7,12 Hexadecane (5.0 g, 12.8 mmol), furan-2-ylmethanamine (1.4 g, 14.1 mmol), potassium iodide (2.1 g, 12.8 mmol), potassium carbonate (3.5 g, 25.6 mmol), and acetonitrile (100 mL) were added to a single-mouth round-bottom flask and reacted at a constant temperature of 50°C for 24 hours. After the reaction was completed, the insoluble matter was filtered, the organic phase was concentrated, and the white solid powder (3.4 g, 65.2%) was purified by column chromatography. 1 H NMR(CDCl3 500MHz) δ7.75(s,1H),6.57(s,1H),6.52(s,1H),5.89(d,1H),5.21(s,1H),4.13(m,2H),3.86(m,2H),3.36(s,2H),2.97-0.89(m,21H).ESI-MS m / z:408.1[M+H] + .

[0099] Example 3, drug efficacy test

[0100] The rat focal cerebral ischemia-reperfusion (tMCAO) model was prepared by suture embolism method, and the therapeutic effect of the test substance on tMCAO model rats was investigated.

[0101] The specific method of establishing the rat focal cerebral ischemia-reperfusion (tMCAO) model is as follows:

[0102] 12 hours before surgery, rats were fed about 30g / rat. Anesthesia was induced with 3-4% isoflurane and maintained with 1.5-2.5% isoflurane. The rats were placed in a supine position on a 37℃ constant temperature operating table. The skin on the midline of the neck was cut open, and the subcutaneous tissue and muscles were bluntly separated. The right common carotid artery, internal carotid artery and external carotid artery were separated. Two ligatures were passed through the external carotid artery, and the distal end was ligated. An artery clamp was placed on each of the common carotid artery and the internal carotid artery. A small cut was made at a 45° angle about 4mm from the bifurcation of the common carotid artery and a suitable thread plug was inserted. The proximal end was gently ligated, and the external carotid artery was cut. The direction and angle of the thread plug were adjusted, and the thread plug was gently pushed. After the thread plug was inserted into the internal carotid artery, the artery clamp was released, and the thread plug was inserted until the black dot mark of the thread plug reached the junction of the internal carotid artery and the common carotid artery, with a length of about 18 to 20mm. The proximal end of the external carotid artery was tied tightly and the suture was fixed. After confirming that there was no bleeding, the skin was sutured, the body temperature of the rat was maintained, and 1 mg / kg of meloxicam analgesic was injected subcutaneously.

[0103] After 2 hours of ischemia, anesthesia was induced with 3-4% isoflurane and maintained with 1.5-2.5% isoflurane. The skin sutures were cut, the ligature at the proximal end of the external carotid artery was opened, the suture plug was removed, reperfusion was performed, and the skin was sutured after re-ligation and confirmation of no bleeding. The sham control group was treated the same as above, except that the suture plug was removed immediately after insertion and no reperfusion was required. The animals could be returned to the cage in prone position after surgery.

[0104] After modeling, when the animals are awake, the animals are scored behaviorally according to Zea Longa's 4-point scoring standard. Animals with scores of 1-3 points are considered to be successfully modeled animals, and animals that do not meet the requirements should be eliminated.

[0105] Table 1. Behavioral scoring table

[0106]

[0107] Methods: SPF male SD rats were used to prepare tMCAO rat models by thread embolization. The animals with successful modeling were randomly divided into 8 groups: model control group, test drug 1, test drug 2, test drug 3, test drug 4, test drug 5, test drug 6 (dihydroartemisinin), test drug 7, with ≥ 6 animals in each group; a sham operation control group was set up, with 6 animals in which the thread embolization was immediately removed after surgery. Each group was given the drug or solvent for the first time 4 hours after cerebral ischemia (4 hours after the thread embolization was removed in the sham operation group), and the dosage of each group was 3×10 - 5mol / kg, the administration volume was 3mL / kg, and the administration was 3 times in total, with an interval of 24h between each administration. Among them, the sham control group and the model control group were catheterized in the right jugular vein to inject the solvent, and the other groups were intravenously injected with each test drug, and the administration time was 20min; the cerebral blood flow was detected before surgery, before the first administration, and 24h after the last administration, and the percentage of cerebral blood flow reduction, recovery, and improvement was calculated; about 24h after the last administration, the latency to fall in the rotarod test, behavioral score, and percentage of cerebral infarction area were detected.

[0108] Solvent: Take 5mL DMSO, 10mL HS-15, 85mL sodium chloride injection (0.9%), mix well to obtain.

[0109] Test drug: Weigh the test substance and dissolve it in the solvent and dilute it to 20 ml.

[0110] Indicators for measuring cerebral ischemia-reperfusion:

[0111] 1. Cerebral blood flow

[0112] Cerebral blood flow was measured in each group before surgery, before the first administration, and 24 hours after the last administration. The last test time was after the behavioral score was scored.

[0113] Table 2. Effects of drugs on cerebral blood flow in rats

[0114]

[0115] Note: vs sham operation control group: ##p<0.01; vs model control group: *p<0.05, **p<0.01;

[0116] 2. Percentage of cerebral infarction area

[0117] After the last cerebral blood flow test, the animals in each group were anesthetized and decapitated, and the brain tissue was quickly removed, the brain was separated, and frozen at -80°C for about 5 minutes. The brain was evenly cut into 5 slices and quickly placed in a 1-2% triphenyltetrazolium chloride (TTC) phosphate buffer solution and incubated at 37°C in the dark until the normal tissue was red and the cerebral infarction tissue was white. The images were taken and the image analysis software ImagePro Plus 6.0 was used to analyze the white tissue area (cerebral infarction area) and the whole brain area. The percentage of cerebral infarction area was calculated according to the formula: percentage of cerebral infarction area (%) = cerebral infarction area / whole brain area × 100%.

[0118] Table 3. Effects of drugs on the percentage of cerebral infarction area in rats

[0119]

[0120]

[0121] Note: vs sham operation control group: ##p<0.01; vs model control group: *p<0.05, **p<0.01.

[0122] 3. Latency to fall in the rat rotarod test

[0123] Within 3 days before surgery, rats were placed on the rotarod instrument and adapted for 5 minutes. During the test, rats were placed on the rotarod instrument and adapted for 10 seconds. The rotarod was started and the speed was set to increase to 25 rpm within 90 seconds. The time when the rats fell (i.e., the latent period of falling) was recorded. If the latent period of falling was greater than 5 minutes, it was counted as 5 minutes. 24 hours after the last administration (i.e., 24 hours after the third administration), the latent period of falling was tested in the same way.

[0124] Table 4 Effects of drugs on the latency to fall in the rat rotarod test

[0125]

[0126] Note: vs sham operation control group: ##p<0.01; vs model control group: **p<0.01; ^, p<0.05.

[0127] 4. Behavioral Scoring

[0128] As shown in Table 5, compared with the sham operation control group, the behavioral scores of rats in the model control group were significantly increased (p<0.01); compared with the model control group, the behavioral scores of rats in the test drug group were significantly decreased (p<0.05 or 0.01).

[0129] Table 5. Effects of drugs on behavioral scores of rats

[0130]

[0131] Note: vs sham operation control group: ##p<0.01; vs model control group: *p<0.05, **p<0.01.

[0132] Each drug group tested could significantly improve the model animals' falling latency, behavioral scores, percentage of cerebral infarction area, and percentage of cerebral blood flow improvement; drug groups 1 and 3 were superior to groups 2, 4, 5, 6, and 7.

[0133] The present invention has been described in detail above. It will be apparent to those skilled in the art that the present invention may be implemented in a wide range under equivalent parameters, concentrations and conditions without departing from the spirit and scope of the present invention and without the need for unnecessary experimentation. Although the present invention provides specific embodiments, it should be understood that further improvements may be made to the present invention. In short, according to the principles of the present invention, this application intends to include any changes, uses or improvements to the present invention, including changes made by conventional techniques known in the art that depart from the scope disclosed in this application. Applications of some of the basic features may be made within the scope of the following appended claims.

Claims

1. A compound represented by formula (I) or its diastereomer, pharmaceutically acceptable salt, ester, or solvate: In formula (I) represents an anion generated by ionization of an inorganic acid or an organic acid; n is any integer between 0 and 5.

2. The compound of formula (I) or its diastereomer, or its pharmaceutically acceptable salt, ester, or solvate according to claim 1, characterized in that: The inorganic acid or organic acid includes at least one of the following: phosphoric acid, sulfuric acid, nitric acid, hydrochloric acid, iodic acid, bromic acid, methanesulfonic acid, maleic acid, fumaric acid, succinic acid, and tartaric acid.

3. A method for preparing a compound of formula (I) according to claim 1 or 2, comprising the following steps: Condensing the compound represented by formula (II) with an amino compound to generate a compound represented by formula (I), wherein the amino compound is trimethylamine or a salt thereof; In the formula (II), Y represents a halogen atom, including Cl, Br or I atom; and n is any integer between 0 and 5.

4. The preparation method according to claim 3, characterized in that: The preparation method of the compound represented by formula (II) is as follows: Compound c is condensed with the compound represented by formula (III) to obtain the compound. The definition of Y in the formula (III) is the same as that of the formula (II); n is any integer between 0 and 5.

5. Use of the compound of formula (I) according to claim 1 or 2 or its diastereomer, or its pharmaceutically acceptable salt, ester, solvate in the preparation of a medicament for preventing and / or treating cerebral stroke or cerebral ischemia-reperfusion injury.

6. The use according to claim 5, characterized in that: The stroke is acute ischemic stroke.

7. The use according to claim 5, characterized in that: The treatment of cerebral ischemia-reperfusion injury includes reducing the cerebral infarction area after ischemia-reperfusion and / or increasing cerebral blood flow.

8. A medicine or pharmaceutical composition for preventing and / or treating cerebral stroke or cerebral ischemia-reperfusion injury, comprising the compound represented by formula (I) according to claim 1 or 2, its diastereomer, or its pharmaceutically acceptable salt ester, solvate, and a pharmaceutically acceptable carrier.

9. The drug or pharmaceutical composition according to claim 8, characterized in that: The stroke is acute ischemic stroke.

10. The drug or pharmaceutical composition according to claim 8, characterized in that: The treatment of cerebral ischemia-reperfusion injury includes reducing the cerebral infarction area after ischemia-reperfusion and / or increasing cerebral blood flow.

Images