CN1194676C - Use of amfetamine alcohol compounds as medicine for treating hepatitis B and preparation method thereof - Google Patents

Abstract

The present invention relates to an application of a phenylalanol compound in preparing medicines for treating hepatitis b and a preparation method of the compound. The active component for resisting hepatitis b viruses of the phenylalanol compound is phenylalanol compound (N-(N-benzoyl-L-phenylalanyl-))-O-actyl-L-phenylalanol, and the compound prepared by separating creeping dichondra herb or prepared by chemical synthesis has the function of resisting hepatitis b viruses. The compound and a compound which is in a dipeptides basic chemical frame structure are not used for preparing medicines for resisting hepatitis b viruses up to now. The compound and the derivatives have obvious suppression on hepatitis b viruses without obvious cytotoxicity, and the compound and the derivatives can be used as medicines for treating patients with hepatitis b.

Description

The phenylalaninol compounds is treated application of hepatitis B medicine and preparation method thereof as preparation

Technical field: the present invention relates to application of the phenylalaninol compounds being treated as preparation the hepatitis B medicine and preparation method thereof, particularly compound N-(N-benzoyl-L-phenylalanyl)-O-acetyl group-L-phenylalaninol is treated the application of hepatitis B medicine and the preparation method of this chemical compound thereof as preparation.

Technical background: hepatitis B is the infectious disease of a kind of serious harm people ' s health of being caused by hepatitis B virus (Hepatitis B Virus), hepatitis B virus causes acute and chronic viral hepatitis B, very high at China's infection rate, it is carriers of hepatitis B surface antigen (HBsAg) that a lot of people are wherein arranged.The persistent infection meeting of hepatitis virus causes liver cirrhosis and primary hepatocyte hepatocarcinoma, and case fatality rate is very high.The medicine of treatment hepatitis B mainly contains the sweet compounds Cymevan of nuclear, acyclovir, model VCV, pressgang pyrimidine at present, Chinese herbal medicine effective ingredients glycyrrhizin, N-methylcantharidimide, polyporusum bellatus, biological species medicine interferon, in addition, some Chinese herbal medicine such as Cacumen Securinegae Suffruticosae, Herba Hedyotidis Diffusae, Spica Prunellae etc. also have certain inhibitory action to hepatitis B virus.Seeking the medicine of specific anti-hepatitis virus from Chinese herbal medicine and ethnic drug, is the urgent task that the medicine and pharmacology interface is faced.

Summary of the invention: when the inventor treats the active component of anti-hepatitis virus in the hepatitis ethnic drug Herba Dichodrae in research, therefrom extraction separation makes a kind of chemical compound, and measure its chemical constitution, find that by the pharmacodynamics screening its certificate has good function of resisting hepatitis B virus, and no cytotoxicity.The inventor finds not see this chemical compound so far and has the medicine of the chemical compound of the basic chemical skeleton structure of this dipeptides as the treatment hepatitis B that the present invention is such:

The chemistry of chemical compound is called N-(N-benzoyl-L-phenylalanyl)-O-acetyl group-L-phenylalaninol, and its chemical constitution is:

Chemical compound of the present invention is produced as follows:

1, from the Chinese herbal medicine Herba Dichodrae, prepare:

The Herba Dichodrae herb is pulverized, and with ethanol merceration twice, filters, and then with 70% ethanol merceration, merge extractive liquid,, concentrating under reduced pressure gets extractum, be suspended in the water after, with petroleum ether extraction three times, collect petroleum ether layer, get cream, silicagel column on the sand cutting behind the concentrating under reduced pressure, with petroleum ether-ether system is eluant gradient elution and centrifugal repeatedly thin layer chromatography, separates obtaining chemical compound.

2, adopt the chemosynthesis mode synthetic: concrete reactions steps is:

(1) be raw material with the L-phenylalanine, thionyl chloride catalysis is descended and methanol carries out esterification reaction of organic acid, gets product 1;

(2) be Reducing agent with the sodium borohydride, in aqueous solution,, get product 2 product 1 reduction;

(3) under-10 ℃ of cryosels baths are cooled off, product 1 is dissolved in the pyridine, drips Benzenecarbonyl chloride. under stirring and react, get product 3;

(4) product 2 is dissolved in the absolute methanol, under the catalysis of Feldalat NM,, gets product 4 with product 3 reactions;

(5) product 4 is dissolved in the anhydrous pyridine, carries out acetylation, get product 5, i.e. the said chemical compound of the present invention with acetic anhydride.

Because the water solublity of the chemical compound that obtains is bad, influence the activity of its anti-hepatitis virus, this chemical compound is carried out structural modification, to improve its water solublity, the derivatives chemical structural formula is as follows:

Derivant 1:R ₁=CH ₂N (CH ₃) ₂HCl (H ₂SO ₄), R ₂=H, R ₃=H, R ₄=CH ₂OAc or CH ₂OH

Derivant 2:R ₁=H, R ₂=NHCH ₂CH ₂N (CH ₃) ₂HCl (H ₂SO ₄), R ₃=H, R ₄=CH ₂OAc or CH ₂OH

Derivant 3:R ₁=H, R ₂=H, R ₃=CH ₂N (CH ₃) ₂HCl (H ₂SO ₄), R ₄=CH ₂OAc or CH ₂OH

Derivant 4:R ₁=H, R ₂=H, R ₃=H, R ₄=COONa or COONH ₄The general formula of this chemical compound and derivant thereof is as follows:

The effect of chemical compound of the present invention and derivant anti-hepatitis virus thereof is:

The chemical compound that is provided has the activity of tangible anti-hepatitis virus, and the no cytotoxicity effect, and this activity is confirmed by following experimental result:

Function of resisting hepatitis B virus:

Materials and methods:

(1) cell in vitro model: HepG2 2.2.1.5

(2) toxicity of mtt assay test sample pair cell

(3) EIA method (HBsAg of Huamei Bio-Engrg Co., and HBeAg diagnostic kit) test sample is to the inhibitory action of HBsAg and HBeAg

(4) positive drug contrast aciclovir (ACV)

Experimental result:

This chemical compound is not seen cytotoxicity when 0.1mol/ml, be 49.0% to the HBsAg suppression ratio, is 20.0% to the HBeAg suppression ratio.The control drug acyclovir is 52.9% to the HBsAg suppression ratio when 0.4mol/ml, is 44.2% to the HBeAg suppression ratio, shows that this chemical compound has the effect that suppresses hepatitis B virus.

Has the activity of anti-hepatitis virus with this dipeptide compounds provided by the invention, make the medicine of treatment hepatitis B, can be used for hepatitis B patient's treatment, by retrieval, do not see this chemical compound so far and have the report of the chemical compound of the basic chemical skeleton structure of this dipeptides as the medicine of treatment hepatitis B.

Embodiments of the invention 1:

Get 4.2Kg Herba Dichodrae herb and pulverize, the industrial alcohol merceration twice with 95% soaked 27 hours at every turn, use filtered through gauze, and then with 70% ethanol merceration 47 hours, merge extractive liquid,, concentrating under reduced pressure gets extractum 350g, after being suspended in the water, use petroleum ether extraction three times, collect petroleum ether layer, get cream 50g behind the concentrating under reduced pressure, silicagel column on the sand cutting, with petroleum ether-ether system be eluant (9: 0.5-3: 7) gradient elution and centrifugal thin layer chromatography repeatedly, separate obtaining this chemical compound 11.7mg.

Chemical constitution to the chemical compound that the present invention obtained is measured:

The white needle, 186～187 ℃ of mp, [α] _D ²⁰-35.71 (0.028, CHCl ₃).IR(KBr)cm ^-1：3314，1725，1661，1631，1533，746，698。EI-MS?m/z：444，384，353，311，269，252，224，172，131，105(10％)，91，77，60。 ¹H-NMR(CDCl ₃，400MHz)：7.69(2H，m，H-3″，7″)，7.50(1H，m，H-5″)，7.42(2H，m，H-4″，6″)，7.0～7.3(10H，m，H-5～9；H-5′～9′)，6.72(1H，d，J＝7.6Hz，NHCOΦ)，5.94(1H，d，J＝8.8Hz，NH-phenylanyl)，4.74(1H，m，H-2)，4.33(1H，m，H-2′)，3.92(1H，dd，J＝11.2Hz，4.0Hz，H-1′)，3.79(1H，dd，J＝11.2Hz，4.0Hz，H-1′)，3.20(1H，dd，J＝13.6Hz，6.0Hz，H-3)，3.03(1H，dd，J＝13.6Hz，6.0Hz，H-3)，2.73(2H，m，H-3′)，2.00(3H，s，CH ₃CO)。 ¹³CNMR(CDCl ₃，100MHz)：170.76(C-1)，170.24(COCH ₃)，167.09(C-1″)，136.67(C-4′)，136.59(C-4)，133.64(C-2″)，131.89(C-5″)，129.27(C-6，8)，129.10(C-6′，8′)，128.73(C-5，9)，128.61(C-5′，9′)，128.55(C-7′)，127.11(C-7)，127.03(C-3″，7″)，126.72(C-4″，6″)，64.56(C-1′)，54.95(C-2)，49.41(C-2′)，38.41(C-3)，37.41(C-3′)，20.77(COCH ₃)。Above spectroscopic data proves that the chemical constitution of this chemical compound is N-(N-benzoyl-L-phenylalanyl)-O-acetyl group-L-phenylalaninol (N-(N-benzoyl-L-phenylalanyl-)-O-actyl-L-phenylalanol).

The product that obtains according to existing pharmaceutical technology, is made into and is can be used for clinical treatment by medicine.

Embodiments of the invention 2:

The chemosynthesis preparation method of N-provided by the invention (N-benzoyl-L-phenylalanyl)-O-acetyl group-L-phenylalaninol is:

(1) with cooling in the bath of 20 ml methanol cryosels, drips 2.4 milliliters of SOCl ₂, adding 4.32 gram L-phenylalanine, stirring at room 2.5 hours refluxed 30 minutes in 80 ℃ of water-baths again, decompression steams methanol, after 14 milliliters of dissolvings of adding methanol, adds 72 milliliters of ether again, cooling, separate out crystallization, sucking filtration gets product 1, i.e. L-phenylalanine methyl ester hydrochloride 5.00 grams;

(2) 769 milligrams of sodium borohydrides are dissolved in 11.2 ml waters, cryosel is bathed cooling, dropping contains 8.6 ml water solution of 1.03 gram phenylalanine methyl ester hydrochlorides, stirring reaction reacted 5 hours under room temperature to room temperature again, reactant liquor equal-volume ethyl acetate extraction 2 times, combined ethyl acetate phase, with the saturated sodium chloride solution washing of 1/3 volume once, ethyl acetate layer with no dried over sodium sulfate after, solvent evaporated gets product 2, i.e. phenylalaninol 0.56 gram;

(3) bathe under the cooling at-10 ℃ of cryosels, 1.07 gram products 1 are dissolved in 5 milliliters of pyridines, stir and drip 0.6 milliliter of Benzenecarbonyl chloride. down, the cryosel bath temperature is increased to room temperature gradually, again in room temperature reaction 5 hours, after reaction finishes, in reactant liquor, add isopyknic water, with equal volume of ethyl acetate once, ethyl acetate layer once removes disacidify with distilled water wash after removing pyridine with the HCl washing of 6M again, the reclaim under reduced pressure ethyl acetate gets product 3, i.e. N-benzoyl-L-phenyalanine methyl ester 1.43 grams;

(4) getting 0.7 gram product 2 is dissolved in 2 milliliters of absolute methanols, cryosel is bathed and is cooled to below-10 ℃, 0.264 milliliter of Feldalat NM-methanol solution of adding 1.74M stirs the methanol solution that adding down contains 1.30 gram products 3, mix homogeneously, reactant liquor moves to refrigerator and cooled and freezes and spend the night, take out next day, reacted 24 hours under room temperature again, and product separates through silica gel column chromatography, chloroform-methanol (20: 1) eluting gets product 4, i.e. N-(N-benzoyl-L-phenylalanyl)-L-phenylalaninol 1.15 grams;

(5) product 4 is dissolved in the anhydrous pyridine, carries out acetylation, get product 5, i.e. N-(N-benzoyl-L-phenylalanyl)-O-acetyl group-L-phenylalaninol with acetic anhydride.

Product 5 and the chemical compound that obtains from Herba Dichodrae are compared, Rf, fusing point, IR, ¹H-NMR, MS unanimity, mixed melting point do not reduce, and prove its chemical constitution.

The product that obtains is become medicine according to the common process making, can be used for clinical treatment.

Claims (2)

1, the phenylalaninol compounds is characterized in that as the application of preparation treatment hepatitis B medicine: with being the medicine that the hepatitis B medicine is treated in preparation, its chemical structural formula is with compound N-(N-benzoyl-L-phenylalanyl)-O-acetyl group-L-phenylalaninol:

2, the preparation method of phenylalaninol compounds as claimed in claim 1 is characterized in that they produce with following method:

Get Herba Dichodrae herb, pulverizing,, filter with ethanol merceration twice; and then with 70% ethanol merceration; merge extractive liquid,, concentrating under reduced pressure gets extractum, is suspended in the water; with petroleum ether extraction three times; collect petroleum ether layer, get cream, silicagel column on the sand cutting behind the concentrating under reduced pressure; with petroleum ether-ether system is eluant gradient elution and centrifugal repeatedly thin layer chromatography, separates obtaining chemical compound: N-(N-benzoyl-L-phenylalanyl)-O-acetyl group-L-phenylalaninol.