CN1526389A - Application of 5,7,4'-substituted flavone in preparing medicine - Google Patents

Abstract

The present invention discloses the new use of 5, 7, 4'-substituted flavone. The 5, 7, 4'-substituted flavone is prepared through extracting Chinese medicinal materials Chinese trumpetcreeper flower, motherwort fruit, privet fruit, schizonepeta herb and Chinese ephedra. It can inhibit obviously the stellate cell proliferation and collagen synthesis of liver has preventing and treating effect on experimental fibration and cirrhosis of liver. It has equivalent dosage obviously lower than that of matrine. The present invention broadens the application fields of 5, 7, 4'-substituted flavone.

Description

The application of 5,7,4 '-replacement flavone in pharmacy

Technical field

The present invention relates to the application of 5,7,4 '-replacement flavone, specifically, relate to 5,7,4 '-replacement flavone and in pharmaceutical field, use.

Background technology

5,7,4 '-replacement flavone is to obtain through chemical modification by extraction in Chinese medicine Flos Campsis, Fructus Leonuri, Fructus Ligustri Lucidi, Herba Schizonepetae, the Herba Ephedrae or by extract, and its general structure is as follows:

R in the formula ₁=H, CH ₃, CH ₃CO or-C ₆H ₁₁O ₅(glucosyl, glucityl);

R ₂=H or-C ₆H ₁₁O ₅(rhammosyl, rhamanopyranosyl).

The present known medical usage of 5,7,4 '-replacement flavone is the application in the preparation antineoplastic agent, and does not appear in the newspapers in the medicine that is applied to prepare treatment or prevent liver cirrhosis.

Summary of the invention

The object of the invention is, the new purposes of 5,7,4 '-replacement flavone is provided, and promptly relates to 5,7,4 '-replacement flavone as the application in the medicine of preparation treatment or prevention liver cirrhosis.

The present invention used 5,7,4 '-replace flavone all to adopt prior art by extracting in Chinese medicine Flos Campsis, Fructus Leonuri, Fructus Ligustri Lucidi, Herba Schizonepetae, the Herba Ephedrae or obtain through chemical modification by extract, concrete extraction and chemical modification method are referring to＜Chinese herbal medicine〉1981; 12 (8): 372, Biochem Syst Ecol 1993; 21 (4): 531, Anal Chem 1965:211:190, Shenyang Pharmacy College's journal 1984; (1): 44, bulletin of Chinese materia medica 1985; 10 (8): 36, R wherein ₁=CH ₃CO, R ₂The substituent of=H is by 5,7, and 4 '-trihydroxyflavone and excess acetyl chloride are separated preparation through silicagel column sephadex LH-20 again.

In order to understand essence of the present invention and convenient explanation better, below with 5,7,4 '-trihydroxyflavone (being commonly called as the celery flavin, apigenin, abbreviation AP) is an example, pharmacological testing and result by it illustrate 5,7, the new purposes of 4 '-trihydroxyflavone in pharmaceutical field.Other replacement chromocor compound all adopts identical pharmacological testing method and identical conclusion is arranged, and as space is limited, does not exemplify one by one at this.Therefore, given example does not limit protection scope of the present invention below.

1, the celery flavin is to the protective effect of SD rat experiment hepatic fibrosis.

(male and female half and half, body weight 164 ± 30g) experimentizes, and tests and establishes negative control group (normal saline), positive controls (matrine), celery flavin (AP) 10mg/kg and 50mg/kg group altogether with Sprague-Dawley (SD) rat.All irritate stomach (ig) administration.

Test method:

SD rat random packet, subcutaneous injection 50% carbon tetrachloride (tetrachloride, CCL ₄)-olive oil 0.1ml/kg, 2 times weekly, preparation rat liver fibrosis model, model control group is only injected olive oil.Negative control group, positive controls and medicine group simultaneously every day the ig normal saline, matrine 50mg/kg and celery flavin 10mg/kg and 50mg/kg.Put to death animals in the 3rd, 6,12 weeks of experiment, get blood prepare determination of serum alanine transaminase (ALT) and hyaluronic acid (hyaluronate, HA); Get liver organization carry out pathologic finding and measure the hepatic tissue hydroxyproline (hydroxyproline, HyP).

Result of the test

(1) to the influence of serum alt, HA and HyP: in the rat liver fibrosis model of tetrachloro-methane induction, serum alt and HA obviously raise during 3 weeks, and HyP is significant change not; ALT, HA and HyP all obviously raise when 6 weeks and 12 weeks.On this basis, observe the influence of AP to ALT, HA and HyP.Found that celery flavin 10 and 50mg/kg and positive controls matrine 50mg/kg all can significantly reduce ALT, HA and the HyP of rising.(table 1).

Table 1 celery flavin is to the influence of alanine transaminase, hyaluronic acid and liver hydroxyproline in the serum

Normal group solvent control group celery flavin celery flavin matrine

(model group) 10mg/kg 50mg/kg 50mg/kg

The 3rd week

ALT(U/L) 157±16 555±178** 425±64 ^Δ 350±28 ^Δ 406±89

HA(ng/ml) 183±17 466±37** 362±76 ^Δ 295±102 ^Δ 334±31 ^ΔΔ

HyP(μg/mg)?1.18±0.20 1.35±0.11 1.36±0.12 1.23±0.12 1.37±0.34

The 6th week

ALT(U/L) 194±58 506±57** 336±28 ^ΔΔ 304±69 ^ΔΔ 374±70 ^ΔΔ

HA(ng/ml) 181±48 451±80** 228±78 ^ΔΔ 191±67 ^ΔΔ 307±59 ^ΔΔ

HyP(μg/mg)?1.23±0.72 4.85 ± 2.92±0.32 ^Δ2.28±0.64 ^Δ3.15±1.03

1.76*

The 12nd week

ALT(U/L) 182±60 881±50** 644±35 ^ΔΔ 605±23 ^ΔΔ 685±58 ^ΔΔ

HA(ng/ml) 169±33 448±78** 211±75 ^ΔΔ 184±82 ^ΔΔ 313±26 ^ΔΔ

HyP(μg/mg)?1.33±0.51 6.28±2.27** 2.89±0.43 ^Δ?2.52±0.55 ^Δ3.35±0.96 ^Δ

Compare with normal group, compare with the solvent control group * * P＜0.01, ^ΔP＜0.05, ^{The Δ Δ}P＜0.01

(2) influence that hepatic pathology is changed: the result of tissue pathology checking demonstration, in the rat liver fibrosis model of tetrachloro-methane induction, a large amount of degeneration of visible hepatocyte, necrosis and inflammatory cell infiltration during 3 weeks; 6 weeks and 12 hepatocellular degeneration when all, necrosis and soak into the collagen fiber hypertrophy in a large number with fibroblast.Celery flavin 10 and 50mg/kg and positive controls matrine all can significantly alleviate rat hepatocytes degeneration, necrosis and connective tissue and form (table 2).

The influence that table 2 celery flavin changes hepatic pathology

The proliferation of fibrous tissue of degeneration necrosis inflammatory cell infiltration

6 all 12 weeks of 3 weeks in 12 weeks in 6 weeks in 3 weeks in 12 weeks in 6 weeks in 3 weeks

Normal group-----～+-～+---

The solvent control group++～++ ++～++ ++ +++-+++～

+++ +++

AP?10mg/kg - -～+ ++ -～+ + + - -～+ +

AP?50mg/kg - -～+ + -～+ -～+ + - -～+ -～+

Matrine--～+ ++-～+++--～++

50mg/kg

2, the celery flavin is to rats'liver sternzellen propagation and the synthetic effect of collagen.

Test method:

The every hole of HSC-T6 cell proliferation test 96 porocyte culture plates adds 1 * 10 ⁴Individual rats'liver sternzellen HSC-T6 is at 37 ℃ of CO ₂Hatch 24h in the incubator, add 100 μ l then and contain the DMEM culture fluid of different pharmaceutical concentration and 10% new-born calf serum (NCS) or do not have medicine culture fluid (quantity of solvent when containing maximum drug level), continue to cultivate the purple staining of 48h post crystallization and measure trap value D (595).The influence of test platelet derived growth factor (PDGF) on cell proliferation then discards cell culture supernatant, adds the culture fluid that contains 0.4%NCS and hatches 48h, adds medicine and PDGF then, hatches 24h again, measures at last and inhales D (595).

The synthetic mensuration of HSC-T6 collagen adopts ³H-proline isotope method.Cell concentration is adjusted into 2.5 * 10 ⁵Individual/ml, every hole adds 100 μ l on 96 orifice plates, cultivates 24h and makes it to form monolayer, to eliminate the cell growth to the synthetic influence of collagen.Add then and contain 50 μ g.ml ^-1Ascorbic medicine and 10%NCS or transforming growth factor ₁(TGF β ₁) cultivate 48h again, add simultaneously every hole [ ³H]-proline 18.5kBq.Cell with trypsinization after, be collected on the glass fiber filter paper, measure cell with liquid scintillation instrument ³H-proline value of mixing (cpm).

Result of the test

(1) the celery flavin is to short HSC-T6 cell proliferation of serum and synthetic celery flavin (6.25～50 μ mol.L that influence of collagen ^-1) the significantly HSC-T6 cell proliferation and the synthetic effect of collagen of the calf serum stimulation of concentration dependence ground inhibition 10% of energy, matrine 2mmol.L ^-1Same inhibitory action (table 3) is also arranged.

Table 3 celery flavin is to the HSC-T6 cell proliferation and the synthetic influence of collagen of serum stimulation

Group cell proliferation/collagen is synthetic/

[n＝6，D(595)] (n＝6，A/cpm)

Solvent control group 1.00 ± 0.05 3089 ± 258

Matrine 2mmol.L ^-10.58 ± 0.07 ^*2172 ± 241 ^*

Celery flavin (μ mol.L ^-1)

6.25 0.98±0.11 3051±247

12.5 0.84±0.02 ^** 2668±314 ^*

25.0 0.72±0.05 ^** 2288±425 ^**

50.0 0.46±0.08 ^** 2114±310 ^**

Compare with the solvent control group, ^*P＜0.05, ^*P＜0.01

(2) the celery flavin is to short HSC-T6 cell proliferation of PDGF and TGF β ₁The short synthetic PDGF 10ng.ml that influences of collagen ^-1Can significantly promote the HSC-T6 cell proliferation, the rate of increase is 42.86%, TGF β ₁2ng.ml ^-1It is synthetic to increase cell collagen, and increasing percentage rate is 86.1% (P＜0.01).Celery flavin (6.25～50 μ mol.L ^-1) can concentration rely on ground inhibition PDGF and TGF β ₁Effect, matrine 2mmol.L ^-1Same inhibitory action (table 4) is also arranged.

Table 4 celery flavin is to short HSC-T6 cell proliferation of platelet derived growth factor and transforming growth factor ₁The short synthetic influence of collagen

Group cell proliferation/collagen is synthetic/

[n＝6，D(595)] (n＝6，A/cpm)

DMEM ^a 0.63±0.03 3688±255

Matched group ^b0.90 ± 0.07 ⁺⁺6717 ± 699 ⁺⁺

Matrine 2mmol.L ^-10.64 ± 0.07 ^*3256 ± 112 ^*

Celery flavin (μ mol.L ^-1)

6.25 0.82±0.06 ^* 6538±484

12.5 0.73±0.04 ^** 5328±159 ^**

25.0 0.64±0.02 ^** 3964±502 ^**

50.0 0.58±0.05 ^** 2909±347 ^**

^a: contain 0.5% NCS, ^b: contain medicine solvent and PDGF (10ng.ml ^-1) or TGF β 1 (2ng.ml ^-1); Compare with the DMEM group, ⁺⁺P＜0.01 is compared with matched group, ^*P＜0.05, ^*P＜0.01

The present invention has following advantage:

(1) the present invention has excavated known compound 5,7,4 '-replace the new medical application of flavones, opened up one New application.

(2) of the present invention 5,7,4 '-replace the flavones safety non-toxic, pharmacological action is strong, indicating well medicinal before Scape.

(3) raw material sources that extract 5,7,4 '-replacement flavone of the present invention enrich, the extractive technique maturation, and can make peroral dosage form or injection type etc., easy to use.

Claims (1)

1,5,7,4 '-replacing flavone, its structural formula is as follows:

R in the formula ₁=H, CH ₃, CH ₃CO or-C ₆H ₁₁O ₅(glucosyl); R ₂=H or-C ₆H ₁₁O ₅(rhammosyl)

Application in the medicine of preparation treatment or prevention liver cirrhosis.