CN103193857A - Holarrhine alkaloid derivative and application of holarrhine alkaloid derivative - Google Patents

Abstract

The invention discloses a holarrhine alkaloid derivative and an application of holarrhine alkaloid derivative. The structure of the holarrhine alkaloid derivative disclosed by the invention is as general formula (I), wherein R represents substituted phenyl or substituted naphthyl and specifically represents the following groups: phenyl, paranitrophenyl, p-methyl phenyl, p-methoxy phenyl, 2,4-dichlorophenyl, p-chlorophenyl, 2,4-dinitrophenyl, 2-naphthyl, 1-naphthyl, 4-chlorine-1-naphthyl and methoxy phenyl. The invention further discloses an application of the holarrhine alkaloid derivative in preparation of a medicine for preventing and treating alzheimer's disease or a medicine for improving the intelligence.

Description

Holarrhine alkaloid derivative and uses thereof

Technical field

The invention belongs to pharmaceutical chemistry and pharmacotherapeutics field, be specifically related to the synthetic and purposes in preparation prevention, treatment senile dementia disease drug or raising intelligence medicine of the holarrhine alkaloid derivative of N-3 replacement.

Background technology

Alzheimer's disease (Alzheimer 2.Isease, namely senile dementia is called for short AD) be a kind of chronic progressive external brain degenerative disease, be core symptom clinically with the cognitive decrease.

Sickness rate and the age of AD are proportionate, and are the main types of senile dementia.

In recent years, clinical study both at home and abroad shows that acetylcholinesterase depressant is the most effective medicine for the treatment of AD, in addition to the treatment of diseases such as glaucoma, myasthenia gravis, intestinal obstruction also have better action (Jin Youyu. pharmacology (the 5th edition), 2001; 54-55.).

Acetylcholinesterase depressant (the acetylocholinesterase inhibitor of FDA approval at present, ACEhI) tacrine (tacrine) E2020 (donepezil), lycoremine (galanthamine) and selagine are arranged, this class medicine is by the activity of acetylcholine esterase inhibition, improve levels of acetylcholine, the function of enhancing maincenter vagusstoff nerve is improved the state of an illness of AD early stage patient to a certain extent.

But still there is certain defective in above-mentioned inhibitor, mainly be bigger to the toxicity of liver, there is in the side effect of nausea and vomiting and the body vagusstoff availability low etc., so Development of New Generation pharmacologically active height, the acetylcholinesterase depressant that toxic side effect is little seems very necessary as the medicine for the treatment of AD.

We find holarrhine alkaloid when screening is used for the acetylcholinesterase depressant of curing senile dementia from plant milk extract: conessine ( 1), isoconessimine ( 2), conessimine ( 3), conarrhimin( 4), conessimine ( 5) (structure as shown in the formula) have inhibiting activity of acetylcholinesterase, preferably, can be used for the treatment of the senile dementia disease drug preparation (Yang Zhongduo etc., the effective monomer preparation method and its usage of tellicherry bark total alkaloids extract, ZL201110074795.7)

The structure of the acetylcholinesterase depressant that separates in the tellicherry bark

Though above-mentioned tellicherry bark total alkaloids has certain acetylcholinesteraseinhibition inhibition, can be for the preparation of the anti-senile dementia disease drug, a little less than the medicine that uses on the market such as selagine effect, and dosage is big.

The invention provides the holarrhine alkaloid that serial N-3 position replaces, as potent vagusstoff anticholinesterase, for the preparation of senile dementia disease drug or raising intelligence medicine.

Summary of the invention

One of purpose of the present invention just provides holarrhine alkaloid and the synthetic method thereof that a series of N-3 position replaces; Another object of the present invention is to disclose described derivative as the purposes of potent acetylcholinesterase depressant at preparation senile dementia disease drug or raising intelligence medicine.

For achieving the above object, the invention provides the holarrhine alkaloid derivative that N-3 that following chemical structural formula I represents replaces.

  

Wherein R is substituted-phenyl, or substituted naphthyl, be specially following group: phenyl, p-nitrophenyl, p-methylphenyl, p-methoxyphenyl, 2,4 dichlorophenyls, rubigan, 2,4 dinitrophenyls, 2-naphthyl, 1-naphthyl, 4-chloro-1-naphthyl, o-methoxyphenyl.

According to a further aspect in the invention, provide a kind of method for preparing the holarrhine alkaloid derivative of the N-3 replacement shown in the chemical formula I, the step that this method comprises and dependency structure formula are as follows:

A, with the lead compound conessine ( 1) at hydrogen peroxide oxidation and Fe ₂SO ₄Reduction changes N-3 demethylation isoconessimine (2) into;

B, phenolic compound and glycol dibromide carry out nucleophilic substitution reaction, obtain compound 6a- 6k,

C, compound 2With compound 6a- 6kBack flow reaction in the presence of the salt of wormwood in acetonitrile makes the holarrhine alkaloid derivative that N-3 replaces 7a- 7k

The holarrhine alkaloid derivative that N-3 provided by the invention replaces 7a-7kBe the inhibitor of potent acetylcholinesterase, can be used as the preparation that effective constituent is used for the senile dementia disease drug or improves the intelligence medicine.

The holarrhine alkaloid that the N-3 that the invention provides replaces 7a-7kAs the application of medicine, can be injection, tablet, pill, capsule, suspended emulsion agent or emulsion etc.

The holarrhine alkaloid derivative that N-3 provided by the invention replaces has the following advantages:

1. in the external biological active testing, the holarrhine alkaloid derivative that 3-N provided by the invention replaces has potent inhibiting activity of acetylcholinesterase,, and wherein the part derivative suppresses the specific activity parent compound 2High 100-1000 times, near the level of positive drug selagine.

2. in vivo in the biological activity test, part holarrhine alkaloid derivative is significantly improved to the ability of learning and memory of drug-induced dementia mice, and effect is near the positive drug selagine.

Embodiment

The present invention is described by the following specific embodiments, can better understand the present invention by specific embodiment, but scope of the present invention is not subjected to the restriction of these embodiment.

Embodiment 1: conessine (compound 1) preparation

Tellicherry bark (powder) 2kg that pulverizes is packed in the 20L round-bottomed flask, use 13L 90% alcohol reflux 3h then, filter, filter residue is used 12L 90% alcohol reflux 2h again, filter, merges twice filtrate and concentrated.

With gained solid substance water dissolution, change over to then in the 5L beaker, transfer PH to 2 with 4M HCl, the static after-filtration that spends the night is transferred PH to 11 with 4M NaOH again, uses isopyknic chloroform extraction then, and underpressure distillation gets tellicherry bark total alkaloids 200g.

With heavily steaming ethanol with tellicherry bark total alkaloids heating for dissolving, carry out separation and purification with macroporous resin then.

Aqueous ethanolic solution with 20%, 30%, 40%, 50%, 60%, 70%, 80% carries out gradient elution.

To contain conessine (compound 1) component merge, carry out separation and purification repeatedly with silicagel column then, eluent is chloroform: methyl alcohol: ammoniacal liquor=15:1:0.1, final conessine (compound 1) 3g.

Conessine (compound 1),The white powder solid, colourless plate crystal, mp 127.0-128.5 ℃; Show orange red with Dragendorff reagent.

ESI-MS m/z:357[M+H] ⁺. ¹H-NMR (400 MHz, CDCl ₃) δ ppm:0.93 (3H, s, H-19), 1.04 (3H, d, J=5.2 Hz, H-21), 1.93 (1H, m, H-18a), 2.21 (3H, s, H-22), 2.33 (6H, s, H-23,24), 2.98 (1H, d, J=10.7 Hz, H-18b), 5.34 (1H, Br s, H-6); ¹³C-NMR (125 MHz, CDCl ₃) δ ppm:38.3 (C-1), 25.1 (C-2), 64.9 (C-3), 35.1 (C-4), 141.8 (C-5), 120.8 (C-6), 32.0 (C-7), 33.4 (C-8), 49.9 (C-9), 36.9 (C-10), 22.0 (C-11), 38.7 (C-12), 50.4 (C-13), 55.8 (C-14), 24.6 (C-15), 27.6 (C-16), 53.5 (C-17), 64.6 (C-18), 19.4 (C-19), 63.2 (C-20), 14.6 (C-21), 41.2 (C-22), 41.6 (C-23), 41.6 (C-24); The literature value of above spectroscopic data and conessine (Zirihi G N, et al. Bioorganic ﹠amp; Medicinal Chemistry Letters, 2005,15:2637-2640) unanimity.

Embodiment 2: holarrhine alkaloid N-3 demethylation derivative (compound 2) preparation

A goes on foot reaction:

With 2g(6mmol) dry holarrhine alkaloid (compound 1) add in the 50mL round-bottomed flask, with the dissolving of 20mL anhydrous methanol, Dropwise 5 mL30% hydrogen peroxide at room temperature reacts and spends the night then.

Add 10mL distilled water afterwards in the reaction solution, stir, use chloroform extraction 3 times, extraction liquid concentrates with the dried over sodium sulfate after-filtration, and purification by silica gel column chromatography gets compound 13 N oxide compounds.

B goes on foot reaction:

With above-claimed cpd 13 N oxide compounds join in the 50mL round-bottomed flask, with the dissolving of 20mL anhydrous methanol, add FeSO ₄7H ₂0 3.3g(12mmol), the nitrogen protection reaction is spent the night under the normal temperature.

In reaction solution, add the saturated NaHCO of 10mL then ₃, add 10mL distilled water behind the reaction 20min, use chloroform extraction 3 times, the extraction liquid dried over sodium sulfate is filtered, and concentrates.

Purification by silica gel column chromatography gets compound 2(954mg)

Compound 2(Isoconessimine ),The white powder solid, shows orange red with Dragendorff reagent by mp 70-72 ℃.

ESI-MS m/z:343[M+H] ⁺. ¹H-NMR (400 MHz, CDCl ₃) δ ppm:0.94 (3H, s, H-19), 1.04 (3H, d, J=5.2 Hz, H-21), 2.21 (3H, s, H-22), 2.33 (3H, s, H-23), 2.98 (1H, d, J=10.2 Hz, H-18b), 5.36 (1H, Br s, H-6); ¹³C-NMR (125 MHz, CDCl ₃) δ ppm:38.7 (C-1), 28.9 (C-2), 59.8 (C-3), 37.9 (C-4), 141.4 (C-5), 120.7 (C-6), 32.0 (C-7), 33.4 (C-8), 49.9 (C-9), 37.1 (C-10), 22.0 (C-11), 39.5 (C-12), 50.3 (C-13), 55.8 (C-14), 24.5 (C-15), 27.6 (C-16), 53.5 (C-17), 64.6 (C-18), 19.4 (C-19), 63.1 (C-20), 14.8 (C-21), 41.2 (C-22), 33.4 (C-23); Above spectroscopic data and isoconessimine (isoconessimine) literature value (Zirihi G N, et al. Bioorganic ﹠amp; Medicinal Chemistry Letters, 2005,15:2637-2640) unanimity.

The preparation of embodiment 3:2-bromotrifluoromethane phenolic ether 6a-6k.

With 0.01mol phenol, 0.5g (0.012mol) NaOH, 0.05g TBAB, 0.05g KI, add in the lump in the there-necked flask, add 20mL distilled water.

Heating is dissolved raw material fully.

When temperature reaches 90 ℃, begin to drip glycol dibromide 1.3mL(0.015mol), control is reflected at about 100 ℃, reaction 5h.

After reaction finishes, cooling, water layer is removed, and organic layer adds 2% NaOH solution washing 2 times, saturated sodium-chloride washing 1 time, dried over sodium sulfate is filtered, and concentrates.

Purification by silica gel column chromatography.

2-bromotrifluoromethane phenolic ether 6a-6k (productive rate 80%---90%).

Embodiment 4: the holarrhine alkaloid derivative that preparation N-3 replaces 7a-7k

With 30mg(0.09mmol) compound 2, the 2-bromotrifluoromethane phenolic ether 6a-6k of 0.24mmol, the K of 0.45mmol ₂CO ₃, the KI of 2mg places the 25ml round-bottomed flask, with the dissolving of 10mL anhydrous acetonitrile, and 85 ℃ of reaction 12h, reaction finishes the back cooling, filters, and concentrates.

Purification by silica gel column chromatography gets compound 7a-7k(productive rate 70%---90%).

Compound The spectral data of 7a-7k is as follows:

Compound 7a( N-benzene oxygen ethyl isoconessimine), colourless plate crystal.

¹HNMR(CDCl ₃, 400 MHz): ESI-MS m/z:463.4[M+H] ⁺. ¹H-NMR (400 MHz, CDCl ₃) δ ppm:0.93 (3H, s), 1.06 (3H, d, J=6.4 Hz), 2.20 (3H, s, N-CH ₃), 2.40 (3H, s, N-CH ₃), 3.02 (1H, d, J=10.4 Hz), 2.89 (2H, t, J=6.4 Hz), 4.04 (2H, t, J=6.4 Hz), 5.35 (1H, Br s), 6.93 (3H, m), 7.29 (2H, t, J=8.4 Hz); ¹³C-NMR (125 MHz, CDCl ₃) δ ppm:38.5,27.4,55.7,34.6,141.8,120.7,31.9,33.4,41.1,37.0,19.5,38.5,50.5,53.4,24.4,24.7,49.9,66.5,19.5,64.2,14.6,38.9,64.2,66.5,52.3, (114.5 2 C), 129.3 (2 C), 141.8,158.8.

Compound 7b( N-(4-oil of mirbane) oxygen ethyl isoconessimine), pale yellow powder shape solid.

ESI-MS m/z:508.2[M+H] ⁺. ¹H-NMR (400 MHz, CDCl ₃) δ ppm:0.93 (3H, s), 1.15 (3H, d, J=6.4 Hz), 2.46 (6H, s, N-CH ₃), 3.18 (1H, d, J=10.4 Hz), 2.93 (2H, t, J=6.4 Hz), 4.14 (2H, t, J=6.4 Hz), 5.35 (1H, Br s), 6.96 (2H, d, J=8.0 Hz), 8.19 (2H, d, J=8.0Hz); ¹³C-NMR (125 MHz, CDCl ₃) δ ppm:38.4,29.0,55.6,34.6,141.4,120.8,31.9,33.5,41.2,37.0,22.0,38.4,50.8,53.4,24.8,24.9,49.8,67.7,19.6,64.3,14.5,39.0,64.3,67.7,52.0, (114.4 2 C), 125.9 (2 C), 141.6,163.9.

Compound 7c( N-(4-methylbenzene) oxygen ethyl isoconessimine), white powder solid.

ESI-MS m/z:477.3[M+H] ⁺. ¹H-NMR (400 MHz, CDCl ₃) δ ppm:0.93 (3H, s), 1.05 (3H, d, J=6.0 Hz), 2.44 (3H, s, N-CH ₃), 2.28 (3H, s, N-CH ₃), 2.22 (3H, s, aromatic ring CH ₃), 3.00 (1H, d, J=10.4 Hz), 2.87 (2H, t, J=6.0 Hz), 4.01 (2H, t, J=6.0 Hz), 5.35 (1H, Br s), 6.80 (2H, d, J=8.0 Hz), 7.06 (2H, d, J=8.0Hz); ¹³C-NMR (125 MHz, CDCl ₃) δ ppm:38.7,27.6,55.8,34.6,141.9,120.7,32.0,33.5,41.2,37.0,22.0,38.4,50.4,53.5,24.5,24.7,50.0,64.6,19.5,64.2,14.8,38.4,63.2,66.7,52.3, (114.3 2 C), 129.8 (2 C), 141.9,156.7.

Compound 7d( N-(4-anisole) oxygen ethyl isoconessimine), white powder solid.

ESI-MS m/z:493.4[M+H] ⁺. ¹H-NMR (400 MHz, CDCl ₃) δ ppm:0.93 (3H, s), 1.04 (3H, d, J=6.0 Hz), 2.43 (3H, s, N-CH ₃), 2.21 (3H, s, N-CH ₃), 3.76 (3H, s, aromatic ring OCH ₃), 2.99 (1H, d, J=10.4 Hz), 2.92 (2H, t, J=6.0 Hz), 3.99 (2H, t, J=6.0 Hz), 5.35 (1H, Br s), 6.82 (4H, m); ¹³C-NMR (125 MHz, CDCl ₃) δ ppm:38.6,27.5,55.7,34.6,141.9,120.7,32.0,33.5,41.1,37.0,22.0,38.4,50.5,53.5,24.5,24.7,50.0,64.4,19.5,64.2,14.7,38.9,63.3,55.8 (OMe), 67.3,52.4,114.5 (2 C), 120.7 (2 C), 153.7,153.0.

Compound 7e( N-(2,4 dichloro benzene) oxygen ethyl isoconessimine), yellow oily liquid.

ESI-MS?m/z:?531.2[M+H] ⁺.? ¹H-NMR?(400?MHz,?CDCl ₃)?δppm:?0.94?(3H,?s),?1.04?(3H,?d,? J=6.4?Hz),?2.43?(3H,?s,?N-CH ₃),?2.21?(3H,?s,?N-CH ₃),?2.99?(1H,?d,? J=10.4?Hz),?2.93?(2H,?t,? J=6.0?Hz),?4.06?(2H,?t,? J=6.0?Hz),?5.35?(1H,?Br?s),?6.84?(1H,?d,? J=8.0?Hz),?7.16?(1H,?d,? J=8.0?Hz),?7.34?(1H,?d,? J=2.0?Hz); ¹³C-NMR?(125?MHz,?CDCl ₃)?δppm:?38.7,?27.6,?55.8,?34.8,?141.8,?120.8,?32.0,?33.5,?41.1,?37.0,?22.0,?38.4,?50.4,?53.5,?24.5,?24.8,?50.0,?64.6,?19.5,?64.3,?14.8,?39.2,?63.2,?68.7,?52.1,?113.9,?123.6,?125.6,?127.5,?129.9,?153.3.

Compound 7f( N-(4-chlorobenzene oxygen) ethyl isoconessimine), colourless block crystallization.

ESI-MS m/z:497.3[M+H] ⁺. ¹H-NMR (400 MHz, CDCl ₃) δ ppm:0.93 (3H, s), 1.04 (3H, d, J=6.0 Hz), 2.42 (3H, s, N-CH ₃), 2.22 (3H, s, N-CH ₃), 2.99 (1H, d, J=10.4 Hz), 2.88 (2H, t, J=6.0 Hz), 4.00 (2H, t, J=6.0 Hz), 5.35 (1H, Br s), 6.82 (2H, d, J=8.8 Hz), 7.21 (1H, d, J=8.8 Hz); ¹³C-NMR (125 MHz, CDCl ₃) δ ppm:38.7,27.6,55.8,34.6,141.8,120.8,32.0,33.5,41.1,37.0,22.0,38.4,50.4,53.5,24.5,24.7,50.0,64.5,19.5,64.2,14.8,38.9,63.2,67.1,52.2, (115.7 2 C), 129.2 (2 C), 125.4,157.4.

Compound 7g( N-(2,4-dinitrobenzene) oxygen ethyl isoconessimine), yellow powder shape solid.

ESI-MS?m/z:?553.4[M+H] ⁺.? ¹H-NMR?(400?MHz,?CDCl ₃)?δppm:?0.93?(3H,?s),?1.05?(3H,?d,? J=6.0?Hz),?2.37?(3H,?s,?N-CH ₃),?2.25?(3H,?s,?N-CH ₃),?3.12?(1H,?d,? J=10.4?Hz),?2.97?(2H,?t,? J=6.0?Hz),?4.27?(2H,?t,? J=6.0?Hz),?5.35?(1H,?Br?s),?7.23?(1H,?d,? J=8.8?Hz),?8.42?(1H,?d,? J=8.8?Hz),?8.76?(1H,?d,? J=2.0?Hz); ¹³C-NMR?(125?MHz,?CDCl ₃)?δppm:?38.3,?27.3,?55.6,?34.7,?141.5,?120.9,?32.0,?33.5,?41.1,?37.0,?22.0,?38.3,?50.7,?53.4,?24.1,?24.8,?49.9,?64.4,?19.5,?63.8,?14.2,?39.0,?63.8,?70.3,?51.9,?114.5,?121.9,?129.0,?134.2,?140.0,?157.2.

Compound 7h( N-(2-naphthalene) oxygen ethyl isoconessimine), white powder solid.

ESI-MS?m/z:?513.2[M+H] ⁺.? ¹H-NMR?(400?MHz,?CDCl ₃)?δppm:?0.94?(3H,?s),?1.19?(3H,?d,? J=6.4?Hz),?2.44?(3H,?s,?N-CH ₃),?2.28?(3H,?s,?N-CH ₃),?3.21?(1H,?d,? J=10.4?Hz),?2.97?(2H,?t,? J=6.0?Hz),?4.18?(2H,?t,? J=6.0?Hz),?5.35?(1H,?Br?s),?7.14?(1H,?s),?7.15?(1H,?d,? J=8.0?Hz),?7.32?(1H,?t,? J=8.0?Hz),?7.43?(1H,?t,? J=8.0?Hz),?7.70-7.76(3H,?m);? ¹³C-NMR?(125?MHz,?CDCl ₃)?δppm:?38.4,?27.1,?55.5,?34.6,?141.7,?120.6,?31.9,?33.5,?41.1,?37.0,?22.0,?38.1,?50.9,?53.3,?23.9,?24.8,?49.8,?64.1,?19.5,?63.3,?13.9,?38.9,?63.3,?66.6,?52.3,?106.6,?119.0,?123.5,?126.3,?126.7,?127.6,?128.9,?129.3,?134.5,?156.8.

Compound 7i( N-(1-naphthalene) oxygen ethyl isoconessimine), white powder solid.

ESI-MS?m/z:?513.2[M+H] ⁺.? ¹H-NMR?(400?MHz,?CDCl ₃)?δppm:?0.95?(3H,?s),?1.04?(3H,?d,? J=6.0?Hz),?2.45?(3H,?s,?N-CH ₃),?2.22?(3H,?s,?N-CH ₃),?3.00?(1H,?d,? J=10.4?Hz),?3.06?(2H,?t,? J=6.0?Hz),?4.21?(2H,?t,? J=6.0?Hz),?5.37?(1H,?Br?s),?6.80?(1H,?d,? J=8.0?Hz),?7.34-7.49?(4H,?m),?7.78?(1H,?d,? J=8.0?Hz),?8.26?(1H,?d,? J=8.0?Hz);? ¹³C-NMR?(125?MHz,?CDCl ₃)?δppm:?38.6,?27.5,?55.7,?34.8,?141.8,?120.8,?32.0,?33.4,?41.1,?37.0,?22.0,?38.4,?50.5,?53.5,?24.4,?24.9,?49.9,?64.4,?19.5,?64.1,?14.6,?39.1,?63.3,?67.2,?52.4,?104.5,?120.1,?122.1,?125.0,?125.6,?125.8,?126.3,?127.3,?134.4,?154.6.

Compound 7j( N-(4-chloro-1-naphthalene) oxygen ethyl isoconessimine), yellow oily liquid.

ESI-MS?m/z:?547.4[M+H] ⁺.? ¹H-NMR?(400?MHz,?CDCl ₃)?δppm:?0.95?(3H,?s),?1.05?(3H,?d,? J=6.0?Hz),?2.47?(3H,?s,?N-CH ₃),?2.23?(3H,?s,?N-CH ₃),?3.00?(1H,?d,? J=10.4?Hz),?3.05?(2H,?t,? J=6.0?Hz),?4.19?(2H,?t,? J=6.0?Hz),?5.36?(1H,?Br?s),?6.71?(1H,?d,? J=8.0?Hz),?7.43?(1H,?d,? J=8.0?Hz),?7.51?(1H,?t,? J=8.0?Hz),?7.60?(1H,?t,? J=8.0?Hz),?8.18?(1H,?d,? J=8.0?Hz),?8.29?(1H,?d,? J=8.0?Hz);? ¹³C-NMR?(125?MHz,?CDCl ₃)?δppm:?38.5,?27.4,?55.7,?34.8,?141.8,?120.8,?32.0,?33.5,?41.1,?37.0,?22.0,?38.4,?50.5,?53.5,?24.3,?24.9,?49.9,?64.2,?19.5,?64.2,?14.5,?39.0,?63.5,?67.5,?52.3,?104.6,?122.5,?123.1,?124.2,?125.7,?125.7,?126.6,?127.4,?131.3,?153.8.

Compound 7k( N-(2-anisole) oxygen ethyl isoconessimine), yellow oily liquid.

ESI-MS?m/z:?493.4[M+H] ⁺.? ¹H-NMR?(400?MHz,?CDCl ₃)?δppm:?0.93?(3H,?s),?1.05?(3H,?d,? J=6.0?Hz),?2.41?(3H,?s,?N-CH ₃),?2.22?(3H,?s,?N-CH ₃),?3.00?(1H,?d,? J=10.4?Hz),?2.96?(2H,?t,? J=6.0?Hz),?4.10?(2H,?t,? J=6.0?Hz),?5.34?(1H,?Br?s),?6.89?(4H,m).? ¹³C-NMR?(125?MHz,?CDCl ₃)?δppm:?38.6,?27.5,?55.8,?34.6,?141.8,?120.7,?32.0,?33.4,?41.1,?37.0,?22.0,?38.4,?50.4,?53.5,?24.4,?24.6,?49.9,?64.4,?19.5,?64.2,?14.7,?38.9,?63.3,?67.3,?52.1,?55.8?(OMe),?111.7,?112.9,?120.8,?121.0,?148.3,?149.3.

Embodiment 5:Ellman method is measured the holarrhine alkaloid derivative 7a-7kAnti-acetylcholinesterase activity

Utilize classical Ellman method (Ellman G.L, Biochemical Pharmacology, 1961; 7:88-95) measured respectively compound 2, 7a-7kTo the half-inhibition concentration of acetylcholinesterase, selagine is as positive control drug.

Concrete experimentation is as follows:

Use 96 hole enzyme plate working samples to the inhibition activity of enzyme, every hole adds the phosphoric acid buffer (pH=8.0) of 140 μ L 0.1M in the sample sets, the sample that adds 20 μ L then, add 160 μ L phosphoric acid buffers in the blank well and do not add any sample, add the selagine of 20 μ L, 0.1 mg/mL and the phosphoric acid buffer of 140 μ L in the positive control control group, the phosphoric acid buffer of the sample of 20 μ L and 155 μ L in the sample copy bottom outlet, other Kong Zhongjun except at the bottom of the sample copy adds the acetylcholinesterase of 15 μ L, mixing is cultivated 20min down at 4 ℃, every hole adds the DTNB(5 of 10 μ L 0.01mM then, 5-dithio two (2-nitrobenzoic acids)) and the ATCI (acetylthiocholine iodide) of 10 μ L, 0.075 mM, mixing is cultivated 20min down at 37 ℃, measures each hole OD value immediately under 405nm.

Every group of experiment triplicate.

And with the inhibiting rate of following formula calculation sample to acetylcholinesterase.

Can calculate sample to the half-inhibition concentration value (IC50) of enzyme according to dose-effect relationship and return law of the straight line.

The result shows that 11 samples are all to the equal strong restraining effect of acetylcholinesterase.

Its IC ₅₀Between 0.11 μ M-4.0 μ M.

Compound wherein 7bActive best, its IC ₅₀Be 110 nM, with the positive drug selagine near (IC ₅₀Be 70 nM).

The results are shown in Table 1.

The inhibiting activity of acetylcholinesterase, of table 1. holarrhine alkaloid derivative 7a-7k

Embodiment 6. Morris water maze laboratories

Healthy Kunming is 150 of male mices, and body weight ± 2g is divided into 15 groups: normal control group, model control group, compound at random 7a-7kTreatment group (0.25 mg/kg), compound 2Treatment group (2 mg/kg) and selagine control group (0.1 mg/kg).

Mouse deducts hair with its head field of operation after anaesthetizing with 1% vetanarcol (80mg/kg), places on the operating table and fixes, and the field of operation iodophor disinfection cuts scalp along sagittal suture, finds the bregma position.

From a bregma left side/right side/1mm, 3mm backward, entry needle inserts 3mm downwards, is hippocampus.

Slowly inject 2.5 μ L condensed state A β 25-35, be 30s inject time, let the acupuncture needle remain at a certain point 30 S, slowly withdrawal of needle.

Sew up a wound.

The instruments that relates in the test, microsyringe etc. all need to sterilize before operation.

Modeling operation back 3d begins gastric infusion, and medicine dissolution-20 ℃ of preservations, thaws before the use in stroke-physiological saline solution.

Normal group and model group are given with volume physiological saline, and beginning Morris water maze study of behaviour is estimated behind the successive administration 14d, reaches the latent period of goal platform in the test 120s.

The result shows, compound 7a-7kAll can obviously shorten the latent period that A β 25-35 causes the platform of reaching home in the dementia mice 120s, namely can both obviously improve the ability of learning and memory of dementia mice, wherein the effect of compound 7b and positive drug selagine is suitable.

And compound 2Can improve the ability of learning and memory of dementia mice to a certain extent, but effect is very weak.

Experimental data sees Table 2

Table 2 compound 7a-7kA β 25-35 is caused the influence of dementia mice Morris water maze space learning memory capability

*: compare P＜0.01, * with model control group: compare P＜0.05 with model control group.

Claims (2)

1. the holarrhine alkaloid derivative that leads to formula I,

Wherein R is substituted-phenyl, or substituted naphthyl, be specially following group: phenyl, p-nitrophenyl, p-methylphenyl, p-methoxyphenyl, 2,4 dichlorophenyls, rubigan, 2,4 dinitrophenyls, 2-naphthyl, 1-naphthyl, 4-chloro-1-naphthyl, o-methoxyphenyl.

2. the described holarrhine alkaloid derivative of claim 1 prevents, treats the purposes in senile dementia disease drug or the raising intelligence medicine in preparation.