Background technology
It is a serious global health problem that chronic HBV (HBV) infects, and the whole world has 3.5 hundred million people to infect hepatitis B virus for a long time approximately according to estimates.And China is the district occurred frequently of viral hepatitis, and the average year sickness rate is about about 2,000,000, has data to show that about at present 1.2 hundred million people of China carry HBV virus.Medicine how to develop the treatment hepatitis B of determined curative effect has become China medicine scholar key task.Clearly emphasize in the Ministry of Science and Technology, State Administration of Traditional Chinese Medicine's key subjects " research of traditional Chinese medical science modernization science and technology development strategy " should " emphasis carries out the traditional Chinese medical science to be duplicated, regulates immunologic function, alleviate hepatocyte injury, improves liver function hepatitis B virus antiviral, especially traditional Chinese medical science anti-hepatic fibrosis, prevent and treat early stage cirrhotic curative effect and Integrated Program Study.”
The treatment of hepatitis B is often based on the antiviral therapy medicine at present, and as interferon, nucleoside analog etc., but curative effect is all unsatisfactory.The antiviral drugs lamivudine that clinical practice is the widest duplicates though it can effectively suppress hepatitis B virus (HBV) fast, and whole curative effect is also well below people's Expected Results.Especially after its side effect, the drug withdrawal relapse rate high and easily cause virus variation disadvantages affect this drug use.Chinese medicine demonstrates comparatively clear superiority by contrast.Experimentation proves, a lot of Chinese herbal medicine or the specially square very curative effect of uniqueness that truly has, for example, Rhizoma Polygoni Cuspidati extract, chlorogenic acid in the Herba Artemisiae Scopariae, coumarin, flavone, polyporusum bellatuss in schisandrin B in matrine in the Radix Sophorae Flavescentis and oxymatrine, the Fructus Schisandrae Chinensis (bifendate), the Polyporus etc. show preferable curative effect at aspects such as antiviral, immunomodulating, hepatic cholagogic, transaminase lowering.But Chinese medicine is bigger because of taking dose, takes effect slowly, and the cycle is long, and the problem of can not fine control medicine effect one-tenth grading is difficult to satisfy people's requirement.
Integrative Chinese-Western is used day by day extensive clinically in recent years, especially at anti-AIDS, tumor, the cardiovascular diseases, influenza, hepatitis, aspects such as gastropathy, its therapeutic effect often is better than single Drug therapy, and SARS storm in 2003 makes the advantage of Chinese medicine and western medicine coupling cause that the world attractes attention, and has also caused that Chinese and western medical circle pays much attention to and unprecedented common recognition simultaneously.National Drug Administration is also to this great attention, and be about to put into effect relevant policies and regulations, the compound preparation that additional Chinese medicine adds the Western medicine joint development is the 4th a kind of new drug of Chinese medicine registration classification the 6th apoplexy due to endogenous wind. therefore, the present invention adopts advanced science and technology to Chinese medicine Rhizoma Polygoni Cuspidati, Herba Artemisiae Scopariae, Fructus Schisandrae Chinensis and Western medicine lamivudine comprehensive development and utilization in addition, adopt combinational therapeutic methods, scientific composition is developed into safe, efficient, controlled, stable, the novel Chinese medicine and western medicine compound preparation easily of treatment hepatitis B diseases with it.
Summary of the invention
The purpose of this invention is to provide a kind of compound preparation for the treatment of chronic viral hepatitis B, this medicine not only has and efficiently suppresses the HBV virus function rapidly, reduce glutamate pyruvate transaminase (ALT) level fast, improve patient's jaundice symptom and hepatic cholagogic, adjusting immunity in addition, reduce the HBV virus variation, recover the generation of liver function and prevention hepatic fibrosis.
Purpose of the present invention can take following method to realize:
A kind of pharmaceutical composition that contains lamivudine is characterized in that it is that component is formed or made active component and pharmaceutically acceptable additives are formed or make active component and pharmaceutically acceptable additives are formed by the alcohol extract of following main materials by the water extract of following main materials by following main materials:
Rhizoma Polygoni Cuspidati 50-300 weight portion, Herba Artemisiae Scopariae 0-300 weight portion, Fructus Schisandrae Chinensis 0-400 weight portion, lamivudine 20-300 weight portion.
It is characterized in that formula optimization is: Rhizoma Polygoni Cuspidati 200 weight portions, Herba Artemisiae Scopariae 150 weight portions, Fructus Schisandrae Chinensis 300 weight portions, lamivudine 100 weight portions.
Its dosage form can be said dosage form on any pharmaceutics, for example capsule, tablet, granule, oral liquid, drop pill, aqueous injection or injectable powder.
Described Fructus Schisandrae Chinensis is schizandrin or deoxyschizandrin.
Described Herba Artemisiae Scopariae is a Herba Artemisiae Scopariae total flavones.
Also comprise polidatin 20-300 weight portion.
Compound preparation of the present invention adopts the composition of medicine of lamivudine and some Chinese medicine extract to treat chronic hepatitis B just.The Chinese medicine extract that the present invention screened is respectively: Rhizoma Polygoni Cuspidati extract, Herba Artemisiae Scopariae extract, Fructus Schisandrae Chinensis extrat.Below be introduction to these materials:
Lamivudine (Lamivudine) is to have the powerful nucleoside medicine of new generation that suppresses the multiple effect of virus.After being used for the treatment of HIV, state Drug Administration and U.S. FDA will be ratified this medicine again in the end of the year 1998 and are used to treat chronic hepatitis B.Its chemistry (2R-cis)-4-amino-1-(2-methylol-1,3-oxygen thia ring penta-5-yl) pyrimid-2-one by name, molecular formula is C
8H
11N
3O
3S; molecular weight is 229; lamivudine can generate the lamivudine triphosphate at HBV infection cell and normal cell intracellular metabolite; the biological activity of the DNA polymerase by reducing the HBV dependenc RNA suppresses the synthetic of HBVDNA, reduces the hepatocellular copy number of getting involved; and by reversing the low reaction state of body T cell; recover the T cytoactive, the protection normal liver cell impels the hepatocellular upset of getting involved simultaneously.Blocking virus DNA's is synthetic, suppresses hbv replication rapidly, and its inhibitory action is sustainable in whole therapeutic process.
Rhizoma Polygoni Cuspidati extract extracts from the Polygonaceae Rhizoma Polygoni Cuspidati, and its effective ingredient mainly is compositions such as anthraquinone class, flavonoid, polydatin, water soluble polysaccharide and tannin.Its anthraquinone analog compound Radix Et Rhizoma Rhei have strong antivirus action; Basic Experiment Study also proves; Rhizoma Polygoni Cuspidati extract all has the good restraining effect to DHB (DHBV) and hepatitis B virus (HBV); and can increase the liver function blood flow; promote hepatocyte to repair and regeneration, reach the effect of protection hepatic injury.Tannin composition wherein can also improve macrophage phagocytic function, the immunoregulation capability of enhancing body.
Herba Artemisiae Scopariae extract extracts from the Chinese medicine Herba Artemisiae Scopariae, effective ingredient mainly contains arcapillin, chlorogenic acid, Quercetin, isorhamnetin and Herba Artemisiae Scopariae water soluble polypeptide etc., Herba Artemisiae Scopariae extract can make the hepatocyte vigor obviously improve, want good hepatoprotective effect, Herba Artemisiae Scopariae can also the quick jaundice eliminating of clearing away heat-damp and promoting diuresis in addition, effectively remove jaundice symptom, the effect of removing that tool is stronger simultaneously and inhibition ultra-oxygen anion free radical, hepatocyte lipid peroxidation capable of blocking, suppress the hepatic tissue fibronectin and express the effect that alleviates degree of hepatic fibrosis.
Fructus Schisandrae Chinensis extrat extracts from the Chinese medicine Fructus Schisandrae Chinensis, and effective ingredient mainly contains schizandrin, deoxyschizandrin, schisandrin B, schisandrin and schisandrol the second grade, and wherein the molecular formula of deoxyschizandrin is C
24H
32O
6Fructus Schisandrae Chinensis extrat has significant hepatocyte injury antagonism, block of the damage of multiple virus to liver plasma membrane, and can effectively reduce glutamate pyruvate transaminase (ALT) level, regulate T lymphocyte differentiation, ripe autoimmune stable to keep, can strengthen liver detoxification function, help the hepatocyte function reparation.
Compound preparation of the present invention designs with the differentiation of tcm theoretical direction, adopt scientific composition, lamivudine is effectively made up with relevant Chinese medicine extract, combine the advantage of Chinese medicine and Western medicine, the effect of performance synergistic corrosion virus remedies the not enough point of single medication again mutually, carry out many target spots at hepatitis B diseases jointly, multi-angle, multi-level treatment.Compound preparation of the present invention has following positive effectiveness: effectively suppress HBV virus, reduce glutamate pyruvate transaminase (ALT) level fast, improve jaundice symptom, hepatic cholagogic, reduce the HBV virus variation, regulate immunity of organism, recover the generation of liver function and prevention and reverse hepatic fibrosis.
The specific embodiment
Describe implementation of the present invention in detail below in conjunction with embodiment.
Embodiment 1:
Rhizoma Polygoni Cuspidati water extract 200mg,
Herba Artemisiae Scopariae water extract 150mg,
Fructus Schisandrae Chinensis water extract 300mg,
Lamivudine 100mg.
Make capsule, tablet, granule, oral liquid etc. by above-mentioned prescription according to the method for producing medicines interpolation proper pharmaceutical excipients of prior art.
Embodiment 2:
Rhizoma Polygoni Cuspidati water extract 200mg,
Fructus Schisandrae Chinensis water extract 300mg,
Lamivudine 100mg.
Make capsule, tablet, granule, oral liquid etc. by above-mentioned prescription according to the method for producing medicines interpolation proper pharmaceutical excipients of prior art.
Embodiment 3:
Rhizoma Polygoni Cuspidati water extract 200mg,
Fructus Schisandrae Chinensis water extract 300mg,
Lamivudine 100mg.
Make capsule, tablet, granule, oral liquid etc. by above-mentioned prescription according to the method for producing medicines interpolation proper pharmaceutical excipients of prior art.
Embodiment 4:
Polidatin 100mg,
Herba Artemisiae Scopariae total flavones 100mg,
Deoxyschizandrin 50mg,
Lamivudine 50mg.
Make lyophilized injectable powder, aqueous injection or oral liquid by above-mentioned prescription according to the method for producing medicines interpolation proper pharmaceutical excipients of prior art.
Embodiment 5:
Polidatin 100mg,
Schizandrin 80mg,
Lamivudine 50mg.
Make lyophilized injectable powder, aqueous injection or oral liquid by above-mentioned prescription according to the method for producing medicines interpolation proper pharmaceutical excipients of prior art.
Embodiment 6:
Rhizoma Polygoni Cuspidati alcohol extract 300mg,
Lamivudine 50mg.
Make capsule, tablet, granule, oral liquid etc. by above-mentioned prescription according to the method for producing medicines interpolation proper pharmaceutical excipients of prior art.
Embodiment 7:
Schizandrin 80mg,
Lamivudine 50mg.
Make capsule, tablet, granule, oral liquid etc. by above-mentioned prescription according to the method for producing medicines interpolation proper pharmaceutical excipients of prior art.
Embodiment 8:
Rhizoma Polygoni Cuspidati alcohol extract 300mg,
Deoxyschizandrin 50mg,
Lamivudine 50mg.
Make capsule, tablet, granule, oral liquid etc. by above-mentioned prescription according to the method for producing medicines interpolation proper pharmaceutical excipients of prior art.
Embodiment 9:
Rhizoma Polygoni Cuspidati alcohol extract 200mg,
Herba Artemisiae Scopariae total flavones 100mg,
Deoxyschizandrin 50mg,
Lamivudine 50mg.
Make capsule, tablet, granule, oral liquid etc. by above-mentioned prescription according to the method for producing medicines interpolation proper pharmaceutical excipients of prior art.
Several main animal experiments contrasts select lamivudine group, Rhizoma Polygoni Cuspidati extract group for use, fill a prescription one group (Rhizoma Polygoni Cuspidati extract+Herba Artemisiae Scopariae extract+Fructus Schisandrae Chinensis extrat+lamivudine) carrying out, its experimental result is as follows:
Experimental example 1:
To CCl
4Cause the influence of Serum ALT in the mouse liver injury, AST
1, experimental technique:
Mice is divided into 5 groups at random, 15 every group.Design lamivudine group, Rhizoma Polygoni Cuspidati extract group, one group of three matched group of prescription are established the dosage group by 10 times of human body recommended amounts, and 0.25g/kg is assigned to desired concn with capsule 's content with distilled water; Establish blank group and CCl in addition
4Positive controls.Five dosage groups are irritated stomach by 0.02ml/g body weight per os and are tried thing; Blank group and CCl
4Positive controls gives distilled water.Animal weighs twice weekly, is tried the agent amount with adjustment, and free diet drinking-water is in testing the 30th day with each treated animal fasting overnight 16 hours.CCl
4Positive controls and each dosage treated animal are irritated the CCl that stomach gives 32mg/kg 1 time by the 0.01ml/g body weight
4Contamination is a solvent with the refined maize oil; The blank group gives Semen Maydis oil 32mg/kg; Respectively be subjected to examination group CCl
4Contaminate and continue to give given the test agent after 4 hours.Give CCl
4Pluck eyeball after 24 hours and get blood, separation of serum is measured ALT, AST, puts to death animal simultaneously.
2, experimental result: CCl
4The ALT of positive controls mice, AST content are than blank group height, and through the T check, difference has significance (P<0.05).ALT value, AST value and CCl that lamivudine group, Rhizoma Polygoni Cuspidati extract group, prescription are one group
4Positive controls more all has decline, and through variance analysis, difference all is significance (P<0.05).Mice is through CCl in this experiment
4After the contamination, CCl
4Hepatocyte large tracts of land necrosis around the matched group central vein, Serum ALT, AST increase greatly, show mice CCl
4The success of hepatocyte injury model copy.Though experimental group hepatic necrosis degree and Serum ALT, AST are higher than the blank group, than CCl
4The matched group pathological changes obviously alleviates, and difference has significance by statistics, illustrate the prescription one group than lamivudine group, Rhizoma Polygoni Cuspidati extract group hepatic injury has more protective effect to the CCl4 induced mice.The results are shown in Table 1.
Table 1 is respectively organized mice ALT, AST assay (x ± s)
| Group | Dosage (g/kg) | Number of animals (only) | ????ALT(U/L) ? | ????AST(U/L) ? |
| The blank group | ??- | ??15 | ??40±14.0* | ??178.5±13.7* |
| Model group | ??- | ??15 | ??1728.7±1277.0 | ??795.1±496.6 |
| Lamivudine group | ??0.06 | ??15 | ??702.5±568.2* | ??293.4±302.0* |
| The Rhizoma Polygoni Cuspidati extract group | ??0.12 | ??15 | ??617.9±695.1* | ??285.9±237.0* |
| Fill a prescription one group | ??0.25 | ??15 | ??568.3±643.3* | ??266.3±252.0* |
* represent and CCl
4Positive controls compares, P<0.05
Experimental example 2:
DHBV is infected the influence of cherry valley duck serum DHBV-DNA:
1, experimental technique: the cherry valley duck duckling is divided into 5 groups at random, 7 every group.Infecting the cherry valley duck duckling with DHBV, after infection the 13rd day was the 5th day (P5) after (T0) before the medication, medication the 7th day (T7), medication the 14th day (T14) and the drug withdrawal, got blood from duck lower limb shin vein, separation of serum, and-70 ℃ of preservations are to be checked.
2, experimental result: cherry valley duck infects back 14 days serum DHBV-DNA total positiveses.In whole experiment, the clear DHBV-DNA level of normal saline matched group Sanguis Anas domestica does not have obvious fluctuation, (P>0.05).1 group of lamivudine group, Rhizoma Polygoni Cuspidati extract group, prescription are still compared with the normal saline matched group before T7, T14 and administration, can both and the significant DHBV-DNA (P<0.001) that reduces; The results are shown in Table 2.
Table 2: to the inhibitory action assay of cherry valley duck DHBV-DNA (mean ± sd)
| Group | Duck is only counted n/ | The A value |
| ???????T0 | ???????T7 | ???????T14 | ????????P5 |
| The blank group | ????7 | ??0.91±0.39 | ??0.90±0.36 | ??0.86±0.31 | ??0.84±0.34 |
| The virus control group | ????7 | ??1.57±0.43 | ??1.19±0.31 | ??1.30±0.37 | ??1.09±0.21 |
| Lamivudine group | ????7 | ??1.67±0.2 | ??0.67±0.23*# | ??0.79±0.07*# | ??0.86±0.23 |
| The Rhizoma Polygoni Cuspidati extract group | ????7 | ??1.53±0.2 | ??0.61±0.32 | ??0.73±0.25 | ??1.00±0.19 |
| Fill a prescription 1 group | ????7 | ??1.62±0.1 | ??0.58±0.49**# | ??0.62±0.45*# | ??0.7±0.62* |
(T0) A value comparison * P<0.05 before different time after the administration (T7, T14) and drug withdrawal (P5) A value (hybridization spot light absorption value) and the administration on the same group, * * P<0.01; Compare with the virus group: #P<0.05, ##P<0.01.
Experimental example 3:
Influence to the peritoneal macrophage phagocytic function:
1, experimental technique: mice is divided into 5 groups at random, 10 every group.Design lamivudine group, Rhizoma Polygoni Cuspidati extract group, one group of three matched group of prescription are established blank group and model control group in addition.The oral tap water 0.2ml/d/ of blank group and model control group is only. and other three experimental grouies are gastric infusion respectively, each organized successive administration 5 days, the 4th, 5 days, other each groups are subcutaneous injection hydroprednisone acetate (prednisolone) 5mg/kg (0.1ml/ only) except that the blank group, and other each groups all at the 4th day lumbar injection sterilization glycogen 2ml/ only except that the blank group.Put to death mice with ethanol intracranial injection method, hank ' the s liquid 2ml for preparing is injected mouse peritoneal, gently rub abdominal part 60 times, make hank ' s liquid thorough washing abdominal cavity,, again abdominal part is cut off an aperture so that obtain more macrophage, draw the interior washing liquid of abdomen in test tube, only stay 0.6ml at last, splash into 2 chicken erythrocyte suspensions and put into 0.4cm length, the wide little glass bar of 0.2cm is in test tube, 37 ℃ of water-baths were cultivated 30 minutes, shook once every 10 minutes, take out glass bar, with PBS washing 3 times, make its natural drying, methanol is fixed 8 minutes, dyes 15 minutes with Ji's nurse Sa dye liquor, with tap water flushing 3 times, behind the natural drying, 100 cells of meter under high power lens, the macrophage number of chicken red blood cell and the chicken red blood cell of being engulfed wherein engulfed in record, calculates phagocytic rate.
2, experimental result: from following table as can be seen, no matter model control group is that phagocytic percentage or phagocytic index all are lower than the blank group, illustrates that the injection hormone can make mouse peritoneal phagocyte phagocytic function descend.
Lamivudine group, Rhizoma Polygoni Cuspidati extract group, one group of three matched group phagocytic percentage of prescription and phagocytic index, all be higher than model control group, data show, lamivudine group, Rhizoma Polygoni Cuspidati extract group, prescription have the effect of the Turnover of Mouse Peritoneal Macrophages of raising phagocytic function for one group. with lamivudine group, Rhizoma Polygoni Cuspidati extract group relatively, one group the nonspecific immunity potentiation of filling a prescription is slightly high.The results are shown in Table 3.
Table 3: to the influence of peritoneal macrophage phagocytic function
| Group | Dosage | Number of animals (only) | Phagocytic percentage X ± SD (%) | Phagocytic index X ± SD |
| The blank group | ??- | ????10 | ????49.4±7.78 | ????0.835±0.186 |
| Model group | ??0.5mg/kg | ????10 | ????19.1±5.26 | ????0.238±0.127 |
| Lamivudine group | ??0.1g/kg | ????10 | ????29.7±6.99 | ????0.334±0.09* |
| The Rhizoma Polygoni Cuspidati extract group | ??0.2g/kg | ????10 | ????35.0±10.9* | ????0.423±0.158* |
| Fill a prescription one group | ??0.25g/kg | ????10 | ????37.0±15.2** | ????0.477±0.295* |
Each group compares with model group: * P<0.05, * * P<0.01