CN111265554A

CN111265554A - Application of antiviral compound composition in resisting HIV virus, SARS virus and leukemia virus

Info

Publication number: CN111265554A
Application number: CN202010212388.7A
Authority: CN
Inventors: 朱寿会; 朱帅
Original assignee: Shenzhen Jinhuiqiu High Tech Co Ltd
Current assignee: Shenzhen Jinhuiqiu High Tech Co Ltd
Priority date: 2020-03-24
Filing date: 2020-03-24
Publication date: 2020-06-12

Abstract

The invention provides an application of an antiviral compound composition in resisting HIV virus, SARS virus and leukemia virus, belonging to the technical field of biological medicine, wherein the antiviral compound composition comprises 7-110g of at least one component for detoxifying, resisting bacteria and diminishing inflammation, 5-60g of at least one component for tonifying qi and nourishing blood and 3-85g of at least one component for tonifying kidney and strengthening yang. The antiviral compound composition provided by the invention can adapt to the variability of viruses through the synergistic effect of multiple components on the premise of no toxic or side effect, and has a remarkable inhibiting effect in the aspect of antivirus.

Description

Application of antiviral compound composition in resisting HIV virus, SARS virus and leukemia virus

Technical Field

The invention belongs to the technical field of biological medicine, and particularly relates to an application of an antiviral compound composition in resisting HIV (human immunodeficiency virus), SARS (severe acute respiratory syndrome) virus and leukemia virus.

Background

The factors causing human diseases are many, wherein the biological factors comprise bacteria, viruses, parasites and the like, half of the diseases are caused by viruses, such as AIDS virus, SARS virus, leukemia virus, human (avian) influenza virus, hepatitis B virus, Ebola virus, papilloma virus of cervical cancer and the like, but the viral diseases are difficult to treat, the spread is fast, the death rate is high, and certain viruses also cause the epidemic all over the world, such as novel coronavirus and influenza virus of the year.

However, because viruses have variability and recognition drug resistance, and western medicines and biological agents cannot adapt to the variation and recognition of the viruses, human beings have unprecedented difficulties in recognizing the viruses and searching for substances for killing the viruses. Chinese traditional medicine through a large amount of laboratory screening, found some Chinese herbal medicine and extracts with antiviral activity, and also demonstrated its antiviral effect in experimental animals, this contains rheum emodin of rhubarb source.

However, emodin is anthraquinone compound, which has poor solubility, is almost insoluble in water, is only soluble in alkali and some organic solvents, and has poor stability and is easy to be oxidized and deteriorated, so that patent applications CN106727482A and CN109528703A both structurally modify emodin, and although the modified emodin has greatly improved solubility, the stability is not improved therewith, and in the face of constantly mutated viruses, the virus inhibition effect which a single component can exert is very limited.

Disclosure of Invention

The invention provides an application of an antiviral compound composition in resisting HIV virus, SARS virus and leukemia virus, the antiviral compound composition is a multi-component compound preparation, can adapt to the variability of the virus by the synergistic effect of the multiple components on the premise of no side effect, and has obvious inhibiting effect in the aspect of antivirus.

In order to achieve the aim, the invention provides an application of a detoxifying and antiviral composition in resisting HIV virus and SARS virus, wherein the composition comprises 7-110g of at least one detoxifying, antibacterial and anti-inflammatory component, 5-60g of at least one qi-tonifying and blood-nourishing component and 3-85g of at least one kidney-tonifying and yang-supporting component.

Preferably, the detoxifying, antibacterial and anti-inflammatory component is at least one selected from honeysuckle, agrimony, liquorice, trichosanthes root, isatis root, houttuynia cordata, forsythia suspensa and anthraquinone extract of rhubarb.

Preferably, the qi-tonifying and blood-nourishing component is at least one selected from fructus Jujubae, radix astragali, rhizoma Polygonati, Atractylodis rhizoma, radix Codonopsis and radix Angelicae sinensis.

Preferably, the kidney-tonifying yang-supporting component is at least one selected from the group consisting of epimedium, acanthopanax, yam, medlar, a temporary erection time, eucommia ulmoides, cistanche deserticola and bupleurum.

Preferably, the composition consists of 4-20 parts of cistanche, 3-30 parts of rhubarb anthraquinone extract, 5-35 parts of houttuynia cordata, 5-30 parts of isatis root, 2-20 parts of radix bupleuri, 5-30 parts of codonopsis pilosula, 3-25 parts of angelica sinensis and 1-15 parts of liquorice.

Preferably, the composition comprises 5-35 parts of rhizoma polygonati, 4-20 parts of eucommia ulmoides, 3-30 parts of rhubarb anthraquinone extract, 5-35 parts of honeysuckle, 1-15 parts of liquorice, 5-30 parts of trichosanthes root and 4-25 parts of the composition before use.

The invention provides an application of an antiviral composition in resisting HIV virus, SARS virus and leukemia virus, wherein the composition comprises 7-110g of at least one component with detoxifying, antibacterial and anti-inflammatory effects, 5-60g of at least one component with effects of benefiting qi and nourishing blood, 3-85g of at least one component with effects of tonifying kidney and supporting yang and 7-75g of at least one component with effects of ventilating lung and relieving cough.

Preferably, the lung-ventilating and cough-relieving component is at least one selected from platycodon grandiflorum, adenophora stricta, fritillaria cirrhosa, rhodiola rosea and ophiopogon japonicus.

Preferably, the composition consists of 3-30g of rhubarb anthraquinone extract, 10-60g of agrimony, 3-25g of rhodiola rosea, 10-30g of astragalus membranaceus, 3-30g of herba epimedii, 5-30 g of radix ophiopogonis, 2-20 g of fritillaria and 5-30 g of fructus ziziphi jujubae.

Preferably, the composition consists of 3-30 parts of rhubarb anthraquinone extract, 4-30 parts of fructus forsythiae, 2-20 parts of platycodon grandiflorum, 5-30 parts of acanthopanax, 10-40 parts of Chinese yam, 5-25 parts of medlar, 5-30 parts of adenophora stricta and 5-25 parts of bighead atractylodes rhizome.

Preferably, the anthraquinone extract of rhubarb comprises at least one of chrysophanol, emodin, physcion, aloe-emodin and rhein.

Preferably, the antiviral compound composition is prepared by the following preparation method:

weighing the components according to the weight, removing impurities, cleaning, drying in the sun, and soaking in 8-20 times of purified water in a container for 0.5-3 days;

putting the soaking solution into a container, boiling with strong fire, keeping with slow fire for 30-600 min, filtering, and concentrating to obtain soft extract;

drying and pulverizing the soft extract, mixing with anthraquinone extract, adding appropriate amount of medicinal adjuvants, and making into pharmaceutically acceptable dosage forms.

Preferably, the dosage form can be capsule, tablet, granule, pill, oral liquid, etc.

Preferably, the dosage of the active ingredients of the antiviral compound composition is 0.1-2 g/kg/d.

Compared with the prior art, the invention has the advantages and positive effects that:

the antiviral composition provided by the invention is a multi-component compound preparation, the safety of all raw materials in the preparation is recorded in Chinese pharmacopoeia of 2005 edition, the compound preparation is scientifically and reasonably matched, the compound preparation can adapt to the variability of viruses through the synergistic effect of multiple components on the premise of no side effect, and pharmacodynamics and toxicology experiments prove that the compound preparation not only has a remarkable inhibiting effect in the aspect of antivirus, but also has the effects of resisting bacteria and diminishing inflammation, improving immunity and shortening treatment course. Furthermore, the compound preparation can be prepared into various dosage forms such as tablets, capsules, pills, granules, pills, oral liquid and the like on the premise of simply changing the dosage form process, and can be conveniently used under various conditions.

Detailed Description

The technical solutions in the embodiments of the present invention will be clearly and completely described below, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.

The embodiment of the invention provides application of an antiviral composition in resisting HIV virus, SARS virus and leukemia virus, wherein the composition comprises 7-110g of at least one component for detoxifying, resisting bacteria and diminishing inflammation, 5-60g of at least one component for tonifying qi and nourishing blood and 3-85g of at least one component for tonifying kidney and supporting yang.

Wherein the antibacterial and anti-inflammatory component is at least one selected from extracts of flos Lonicerae, herba Agrimoniae, Glycyrrhrizae radix, Trichosanthis radix, radix Isatidis, herba Houttuyniae, fructus forsythiae and anthraquinone; the qi-tonifying and blood-nourishing component is at least one selected from fructus Jujubae, radix astragali, rhizoma Polygonati, Atractylodis rhizoma, radix Codonopsis and radix Angelicae sinensis; the kidney invigorating and yang supporting component is selected from at least one of herba Epimedii, radix Acanthopanacis Senticosi, rhizoma Dioscoreae, fructus Lycii, radix Paeoniae alba, Eucommiae cortex, Cistanchis herba and bupleuri radix.

The embodiment of the invention also provides application of an antiviral composition in resisting HIV virus, SARS virus and leukemia virus, wherein the composition comprises 7-110g of at least one component for detoxifying, resisting bacteria and diminishing inflammation, 5-60g of at least one component for tonifying qi and nourishing blood, 3-85g of at least one component for tonifying kidney and yang and 7-75g of at least one component for ventilating lung and relieving cough.

Wherein the detoxifying, antibacterial and anti-inflammatory component is at least one selected from extracts of honeysuckle, agrimony, liquorice, trichosanthes root, isatis root, houttuynia cordata, forsythia and anthraquinone of rhubarb; the qi-tonifying and blood-nourishing component is at least one selected from fructus Jujubae, radix astragali, rhizoma Polygonati, Atractylodis rhizoma, radix Codonopsis and radix Angelicae sinensis; the kidney invigorating and yang supporting component is selected from at least one of herba Epimedii, radix Acanthopanacis Senticosi, rhizoma Dioscoreae, fructus Lycii, radix Paeoniae alba, Eucommiae cortex, Cistanchis herba and bupleuri radix; the lung ventilating and cough relieving component is at least one of radix Platycodi, radix Adenophorae, Bulbus Fritillariae Cirrhosae, radix Rhodiolae and radix Ophiopogonis.

The raw materials in the formula have the following effects:

rhubarb, radix et rhizoma Rhei, is bitter, cold and nontoxic. It enters spleen, stomach, large intestine, liver and pericardium meridians. [ function and indications ] purgation and elimination of accumulation, clearing heat and purging fire, cooling blood and removing toxicity, removing blood stasis and dredging channels, promoting diuresis and eliminating jaundice. Can be used for treating constipation due to excessive heat accumulation, hematemesis, epistaxis, conjunctival congestion, pharyngeal swelling, carbuncle, furuncle, intestinal carbuncle, abdominal pain, blood stasis, amenorrhea, puerperal blood stasis, traumatic injury, damp-heat dysentery, jaundice, dark urine, stranguria, and edema; it can be used for external treatment of burn and scald. The wine rhubarb is good at clearing heat and toxicity in the upper jiao and blood system. It can be used for treating conjunctival congestion, pharyngeal swelling, and gingival swelling and pain. The cooked rhubarb has the functions of relieving the purgation, purging fire and removing toxicity. It can be used for treating pyocutaneous disease due to fire toxin. Rhubarb charcoal cools blood, removes stasis and stops bleeding. Can be used for treating blood heat with blood stasis and hemorrhage.

The agrimony (nature and taste) is smooth, pungent and astringent, cool and nontoxic, and enters heart and liver meridians. [ function and indication ] clear heat and promote diuresis, tonify deficiency and activate blood. It can be used for treating urinary tract infection, leucorrhea, dysentery, weakness after illness, sprain, and contusion.

Rhodiola root (nature and taste and meridian tropism) is flat, sweet, astringent and nontoxic, and enters lung and heart meridians. Has the functions and indications of invigorating vital energy, clearing away lung-heat, promoting intelligence, nourishing heart, arresting bleeding, dissipating blood stasis and eliminating swelling. Can be used for treating qi deficiency, asthenia, aversion to cold, short breath, asthenia, cough due to lung heat, hemoptysis, leucorrhea with diarrhea, and traumatic injury.

The astragalus root (nature and flavor and meridian tropism) is sweet in taste, slightly warm in nature and non-toxic; it enters spleen and lung meridians. [ Functions and indications ] tonify qi, strengthen superficies, expel toxin, expel pus, induce diuresis and promote granulation. Can be used for treating deficiency of vital energy, asthenia, chronic diarrhea, proctoptosis, spontaneous perspiration, edema, uterine prolapse, albuminuria due to chronic nephritis, diabetes, and unhealed wound.

Xianlingpi (nature, flavor and channel tropism) is pungent, warm, non-toxic, and enters liver and kidney channels. [ Functions and indications ] tonifying kidney yang, strengthening bones and muscles, dispelling wind-damp, relieving cough, dispelling phlegm, promoting hematopoiesis and immunity.

Acanthopanax root (nature, taste and channel tropism) is sweet, slightly bitter, warm and nontoxic; it enters spleen, lung, heart and kidney meridians. The composition has effects of invigorating qi, invigorating spleen, invigorating kidney, and tranquilizing mind, and can be used for treating spleen and lung qi deficiency, kidney deficiency, waist and knee pain, heart and spleen deficiency, insomnia, and amnesia.

The fructus Jujubae [ nature, flavor and meridian tropism ] is sweet, warm and nontoxic. It enters spleen and stomach meridians. [ Functions and indications ] tonify the middle-jiao and Qi, nourish blood and tranquilize the mind.

Radix Adenophorae (with nature, flavor and meridian tropism) is sweet, slightly bitter, slightly cold in nature and nontoxic. Meridian tropism: entering the lung; the stomach meridian. [ function and indication ] nourishing yin and clearing heat; moistening lung and eliminating phlegm; benefiting stomach and promoting fluid production. Chronic cough due to yin deficiency; cough with bloody phlegm; dry and cough with little sputum; deficiency-heat pharyngitis; thirst due to body fluid consumption.

Forsythia suspense (property, taste and meridian tropism) is slightly cold, bitter in taste and non-toxic. The method comprises the following steps: lung, heart and small intestine. [ function and indication ] clearing away heat and toxic material, reducing swelling and resolving masses. Belonging to the category of heat-clearing herbs.

The root of balloonflower (nature, flavor and meridian tropism) is bitter, pungent, mild and nontoxic. The method comprises the following steps: the lung meridian. [ Functions and indications ] it is good at dispersing lung qi, eliminating phlegm and relieving sore throat, and also has the action of expelling pus, and is mainly indicated for cough with excessive phlegm, pharyngalgia hoarseness and lung abscess with pus discharge. Disperse lung qi, relieve sore throat, dispel phlegm, and expel pus.

The yam (nature, taste and channel tropism) is sweet, mild and nontoxic. Meridian tropism: it enters spleen, lung and kidney meridians. [ Functions and indications ] tonify spleen and stomach, promote the production of body fluid and benefit lung, tonify kidney and arrest seminal emission. Can be used for treating spleen deficiency, anorexia, chronic diarrhea, lung deficiency, cough, asthma, spermatorrhea, leukorrhagia, frequent micturition, and diabetes due to deficiency heat. The bran-parched rhizoma Dioscoreae has effects of invigorating spleen and invigorating stomach. Can be used for treating spleen deficiency, anorexia, diarrhea, loose stool, and leukorrhagia.

Medlar (nature, taste and meridian tropism) has mild nature, sweet taste and no toxicity. The method comprises the following steps: the liver and kidney meridians. [ Functions and indications ] nourishing liver and kidney, replenishing vital essence and improving eyesight. They belong to the yin tonics classified under the deficiency tonics.

Bighead atractylodes rhizome (with nature, taste and meridian tropism) is warm in nature, sweet in taste, bitter and nontoxic. The method comprises the following steps: spleen meridian and stomach meridian. [ function and indication ] invigorate the spleen, replenish qi, dry dampness and induce diuresis, stop sweating and prevent abortion. They belong to the category of qi tonics under deficiency-tonifying herbs.

Honeysuckle flower (nature and flavor with meridian tropism) is cold in nature and sweet in taste. The method comprises the following steps: lung, heart and stomach meridians. [ function and indication ] clearing away heat and toxic material, and cooling and dispelling wind-heat. Heat-clearing and toxicity-removing herbs belonging to the category of heat-clearing herbs

When used, the original flavor and meridian tropism are slightly warm in nature, sweet and pungent in flavor. The method comprises the following steps: kidney and liver meridians. [ Functions and indications ] tonify kidney yang, strengthen tendons and bones, dispel wind-damp. They belong to yang tonics classified under deficiency tonics.

Licorice root, radix Glycyrrhizae (nature and flavor and meridian tropism) is sweet and neutral in nature and enters heart, lung, spleen and stomach meridians. [ function and main treatment ] invigorating spleen and replenishing qi, eliminating phlegm and relieving cough, relieving spasm and pain, clearing heat and detoxicating, and harmonizing the drugs.

Trichosanthis radix (nature and flavor and meridian tropism) is sweet, slightly bitter and slightly cold. Meridian tropism: it enters lung and stomach meridians. [ function and indication ] clearing heat and promoting the production of body fluid, reducing swelling and discharging pus. Can be used for treating fever polydipsia, lung heat dry cough, internal heat diabetes, pyocutaneous disease and pyogenic infections.

Radix Isatidis (nature, taste and meridian tropism) is bitter and cold. Meridian tropism: the heart and stomach meridians. [ Functions and indications ] clear away heat and toxic material, cool blood and relieve sore throat.

The cordate houttuynia is pungent, slightly cold and non-toxic. Meridian tropism: it enters lung meridian. [ function and indication ] clearing away heat and toxic material, curing carbuncle and discharging pus, inducing diuresis and treating stranguria. Can be used for treating lung abscess with purulent vomiting, phlegm heat, cough and asthma, dysentery, pyretic stranguria, carbuncle, swelling, and sore.

Cistanche deserticola (nature, flavor and channel tropism) is sweet, salty, warm and nontoxic. Meridian tropism: it enters kidney and large intestine meridians. [ Functions and indications ] tonify kidney yang, replenish essence and blood, moisten intestines to relieve constipation. Can be used for treating sexual impotence, infertility, soreness of waist and knees, weakness of tendons and bones, constipation due to intestinal dryness.

Codonopsis pilosula (nature, flavor and channel tropism) is sweet, mild and nontoxic. Meridian tropism: it enters spleen and lung meridians. [ Functions and indications ] tonify the middle-jiao and qi, invigorate the spleen and benefit the lung. Can be used for treating spleen and lung deficiency, short breath, palpitation, anorexia, loose stool, asthma, cough, internal heat, and diabetes.

Dang Gui (property and flavor and meridian tropism) is sweet, pungent and warm. Meridian tropism: the liver, heart and spleen meridians. [ function and indication ] enriching blood and promoting blood circulation, regulating menstruation and relieving pain, and loosening bowel to relieve constipation. Can be used for treating blood deficiency, sallow complexion, giddiness, palpitation, menoxenia, amenorrhea, dysmenorrhea, asthenia cold, abdominal pain, constipation due to intestinal dryness, rheumatic arthralgia, traumatic injury, superficial infection, pyocutaneous disease.

Eucommia bark (nature, taste and meridian tropism) is sweet, warm and nontoxic. Meridian tropism: it enters liver and kidney meridians. [ Functions and indications ] tonify liver and kidney, strengthen tendons and bones, and prevent miscarriage. Can be used for treating lumbago due to kidney deficiency, weakness of bones and muscles, and pregnant bleeding and threatened abortion; hypertension is caused.

In the formula, the components for detoxifying, resisting bacteria and diminishing inflammation are monarch drugs, have the effects of detoxifying, resisting bacteria and diminishing inflammation, are the essential drugs for removing various toxins, the components for tonifying qi and nourishing blood and the components for tonifying kidney and supporting yang are ministerial drugs, are matched with the monarch drugs to play the effects of tonifying qi and strengthening exterior, expelling toxin and expelling pus, and can assist the body to promote the hematopoietic function and improve the immune function. The components are matched to play roles in clearing away heat and toxic materials, supporting toxicity and expelling toxin, promoting hematopoiesis and improving immunity. According to the severity of the patient, the composition can be matched with lung-ventilating and cough-relieving components to relieve the damage to the organism, and particularly, the protection to the lung can be strengthened. The feasible components listed in the components have similar medicinal effects, and only slight difference exists in the strength of the medicinal effect, so that doctors can reasonably match the components according to the actual conditions of patients. The raw materials of the provided antiviral composition are recorded in national standards, have considerable safety, and can prove that the effective components extracted based on the antiviral compound composition have theoretical basis for treating HIV virus, SARS virus and leukemia virus and have obvious inhibiting effect based on in vivo animal models and in vitro cell detection.

In the antiviral composition, the rhubarb anthraquinone extract comprises at least one of chrysophanol, emodin, physcion, aloe-emodin and rhein, and the rhubarb anthraquinone extract can be fluid extract of rhubarb anthraquinone, crushed body of dried rhubarb anthraquinone extract or further prepared particles by crushing.

In addition, the extraction raw material and the extraction method for the rhubarb anthraquinone compound are well known to the technicians in the field, and the principle is as follows: the anthraquinone glycoside is hydrolyzed into aglycone by dilute sulphuric acid solution, and extracted by hot chloroform/ether by utilizing the property that free anthraquinone can be dissolved in hot chloroform/ether. It is understood that there are many extraction methods for implementing the above principle, and the condition parameters can be adaptively adjusted according to different situations. In this regard, the extraction method is not within the scope of the present application, as long as the rhubarb anthraquinone-based extract or the particles thereof can be obtained.

Based on the above principle, the embodiment of the present application provides an existing extraction method, which specifically includes the following steps: 1) slicing radix et rhizoma Rhei, cutting, pulverizing to obtain radix et rhizoma Rhei coarse powder, and sieving with 80 mesh sieve to obtain radix et rhizoma Rhei fine powder; 2) collecting radix et rhizoma Rhei powder 20g, adding 20% H₂SO₄200ml, heating in water bath at 70 ℃ for 3-4 hours; 3) suction filtering, washing filter cake to neutrality, and heating to 70 deg.CDrying and grinding into powder; 4) heating the extractant in the round-bottom flask to volatilize, leading the steam to go upwards through the gas-guide tube, cooling the steam into liquid in the condensing tube, and dripping the liquid into the tube provided with the sample filter paper cylinder to immerse the sample in the pure extractant; the substance to be extracted is gradually dissolved into the extracting agent from the sample; when the liquid level of the extracting agent in the sample tube gradually rises to be level with the upper end of the return tube, the extracted substances are carried to flow into the flask from the siphon tube on the side surface together under the siphon action, and then a round of circulation is completed; drying the filter cake, placing in a Soxhlet extractor, reflux-extracting with diethyl ether about 150ml for 3-4 hours to obtain diethyl ether extract, removing solvent, and volatilizing at room temperature to obtain rhubarb anthraquinone extract.

In order to more clearly and specifically introduce that the antiviral compound composition provided by the embodiment of the invention has effective effects of resisting HIV virus, SARS virus and leukemia virus, the invention takes an antiviral compound composition consisting of 3-30g of rhubarb anthraquinone extract, 10-60g of agrimony, 3-25g of rhodiola rosea, 10-30g of astragalus root, 3-30g of herba epimedii, 5-30 g of radix ophiopogonis, 2-20 parts of fritillaria and 5-30 parts of fructus ziziphi jujubae as an example to carry out the following pharmacodynamics test and toxicity test, so as to show that the antiviral compound composition has no toxic or side effect, but has very remarkable inhibiting effect on resisting HIV virus, L6565MLV leukemia virus and SARS virus. Because the viruses belong to RNA viruses without cell structures and can only be parasitized and propagated in living host cells through replication, the antiviral compound composition provided by the invention can inhibit the viruses by effectively preventing the viruses from being combined with cell membrane receptors and preventing the viruses from being replicated in the cells, but not directly killing the viruses, thereby having remarkable broad-spectrum inhibition effect.

It is understood that the pharmacological effects of other agents based on the inventive concept are not statistically different from the effects of the following experiments, and thus may be understood as the effects are the same or nearly similar, and thus the remaining experiments are not repeated.

1. Pharmacological pharmacodynamics experiment

The pharmacodynamic experiment comprises an in vitro intracellular experiment and an in vivo animal model experiment.

1.1 in vitro intracellular experiments

In vitro intracellular experiment, cytopathic effect (CPE) is observed, TC740 cells are used as target cells, and half Inhibitory Concentration (IC) of the antiviral compound composition on viruses is adopted₅₀) And the Therapeutic Index (TI) as objective indices.

In performing in vitro (intracellular) experiments, the present invention performed comparative experiments for the following two cases.

1.1.1 adding murine leukemia virus L6565MLV into cells in which target cells TC740 normally grow for 90 minutes, adding antiviral compound compositions with different doses, observing the inhibition effect of the extract on HIV, and observing that the HIV loading capacity is obviously reduced along with the increase of the concentration of the extract, wherein the data are shown in Table 1 and good inhibition effect is shown.

TABLE 1 in vitro anti-HIV effect of the extract

By statistical calculation, the antiviral compound composition has half Inhibitory Concentration (IC) on HIV₅₀) 7.6mg/ml and a Therapeutic Index (TI) of 43.8. The result proves that the extract has strong inhibiting effect on HIV and shows dose-effect relationship, and the inhibiting rate is increased along with the increase of the concentration of the extract.

1.1.2 adding the antiviral compound composition into the cells of the target cell TC740 which normally grows, then adding L6565MLV for attacking, and observing that the cells are obviously protected, only about 3-5% of the cells have CPE, and the virus load is reduced. Therefore, the antiviral compound composition is effective to HIV and has strong prevention effect on leukemia virus, and the data are shown in Table 2.

TABLE 2 toxic Effect of the extract on TC740 cells

1.2 in vivo pharmacodynamics

This example uses murine leukemia virus (L6565MLV) instead of HIV, Balb/C mice were modeled as subjects infected with MLV, and then HIV studies were performed in mice with the drug. In order to observe the effect of the drug in mice, this example designed administration control groups (divided into three large, medium and small dose groups), normal control groups and virus infection control groups. Spleen index, blood index, mouse behavior, and MLV nucleic acid content were used as observation indexes.

Statistical results show that the spleen index and blood index of the administration experimental group are different from those of the virus infection group, wherein the two groups with drug doses of 10mg and 15mg in the administration experimental group are significantly different from those of the virus infection group (LP < 0.01).

The content of L6565MLV nucleic acid in a mouse is detected by a semi-quantitative RT-PCR method, the result shows that the ratio of the L6565MLVRIVA target gene amplified under different doses of drugs to the absorption area of β -actin is reduced along with the increase of the drug concentration, which indicates that the virus reproduction is weakened when the drug dose is increased, and the inhibition effect of the drugs on the virus is fully proved, compared with an infected control group, the ratio of a 15mg dose administration experimental group is only (0.88) which is obviously lower than that of an infected group (2.58), and the two groups have obvious difference.

1.3 pharmacological experiments

In the in vitro study of the anti-HIV effect of the drug, in order to explore pharmacology and target points, a special group is also designed in the administration experimental group, namely after TC740 cells grow normally, different doses of the drug are added, and then L6565MLV is added to observe the anti-HIV effect of the drug and compare with other groups.

Based on this particular set of observations, the cells were clearly protected after drug addition, with only a few cells being attacked to develop CPE. This phenomenon is consistent with the administration of HIV after 90 minutes of challenge to the cells in the dosing group. This fact shows that the action of the drug on HIV is expressed in two aspects, namely, the drug can enter cells to play the function of inhibiting virus replication in the cells, and the drug can prevent the HIV from being combined with cell receptors, so that the HIV cannot be fused with cell membranes, and the cells are not attacked. Subsequent researches prove that the target point of the drug generation effect can inhibit the reverse transcriptase activity of HIV, prevent viruses from being adsorbed on cells, directly participate in the prevention of HIV-gp120 binding sites and the like to realize the HIV inhibition. In common pharmacological research, the result shows that the medicament has no harmful effect on the nervous system, the respiratory system and the cardiovascular system of the mouse through the effect of the medicament on the mouse.

2. Toxicology studies

This example illustrates the safety of the provided rhubarb horsetails combination composition based on cytotoxicity, acute toxicity and chronic toxicity.

2.1 cytotoxicity

The cells affected in this example were the culture medium target cells TC740, and the drug concentration was 25mg/ml minimum and 1600mg/ml maximum, and the results showed that different drug doses were toxic to TC740, as the cell survival rate was inversely proportional to the drug concentration (see Table 7). TC of the drug by MTT assay detection and statistical calculation₅₀332.8 mg/ml.

TABLE 7 cytotoxic Effect of drug TC740

2.2 acute toxicity study

The animals used in this example were rats weighing between 100-120g, and half male and half female. The 72 white rats are randomly divided into 6 groups and 12 rats in each group, the 6 groups have the drug dosage of 17000mg, 25000mg, 35000mg, 50000mg, 70000mg and 105000mg per kilogram of body weight respectively, and the administration is performed once a day for 10 days. No death was observed in 25000mg/kg of animals, as confirmed by observation, and drug LD was calculated statistically₅₀It was 77964.3567 mg/kg.

2.3 Long term toxicity Studies

In this example, 101 rats with body weights between 110-120g, and 50 rats, 51 males and females, were randomly divided into 4 groups, three experimental groups (25 rats each) and one control group (26 rats). The drug doses are respectively as follows: the dosage of the large dose group is 0.92g/kg/d, the middle dose group is 0.31g/kg/d, the small dose group is 0.15g/kg/d, the dosages are 80 times, 40 times and 20 times of the dosage of the human, and the administration method is gastric lavage.

Experimental observations include animal performance, food intake, hair, fecal matter, blood indicators, blood biological indicators, mortality. The pathological examination includes heart, liver, stomach, kidney, lung, brain, testis, uterus, etc. The key point of pathological observation is the change of rat specimen and the change under the mirror. All data were statistically processed using the sps 10.0 software.

The results show that the rats do not die in 12 weeks after gastric lavage, the behaviors are normal, the feces are yellow brown, the properties are not abnormal, the indexes of urine in the experimental group are not obviously different from those of the control group (P >0.05), and the indexes of liver function in the experimental group are not obviously different from those of the control group (P > 0.05).

In terms of pathological anatomy, as shown in table 8, there was no significant difference in the body weight index of each organ between the experimental group and the control group. The difference between the pathological changes of the organs is not statistically significant. The animals in the experimental group had 16 lesions, and the animals in the control group had 5 lesions, the incidence of the former was 19.93%, the latter 19%, and there was no statistical difference between the two. The pathological nature is mostly mild cytopathic lesions, and also appears as pulmonary telangiectasia, alveolar septal thickening, bleeding spots in the brain and gliosis (only one animal), bleeding in the renal cortex, superficial inflammation in the stomach. The special lesion is liver cyst found in a rat. The above lesions are all mild pathological changes, while cellular degeneration is a reversible lesion. Therefore, the two groups of lesions are light in nature, and the number of lesion cases between the two groups of lesions is not obviously different. The incidence of organ disease in the two groups of animals is low, which indicates that the drug does not show long-term toxicity within safe dosage.

TABLE 8 incidence of lesions in organs of two groups of animals

As seen from the results in Table 8, the proportion of total lesions was within 20% in both the experimental group and the control group. The total abnormality rate of the experimental group was 19.93%, the control group was 19%, and there was no significant difference between the two. From the specific pathological nature, these lesions are all slight changes, some are reversible lesions (e.g. degeneration of liver cells), and we examined some animals before the experiment and found similar changes, which fully indicates that these changes should be independent of the drug intake. From the diseased organs, the probability of the occurrence of lung and stomach diseases is only slightly high (6.6%) but not more than 10% in the experimental group, and no disease occurs in heart, testis and ovary, which fully indicates that the long-term administration of the medicine has no influence on rats and does not damage the organs. The lesion ratio of the liver was 4% and 3.8% in the experimental group and the control group, respectively, and this ratio was low. The heaviest lesions in both groups of animals were granulomatous changes in hepatocytes in view of the nature of the lesions. Pathological granular pathological changes of the liver are reversible, the pathological changes are mild pathological changes, and ten normal rats are dissected before experiments to find similar pathological changes. The lesion proportion of the lung is 6.6%, the types of the lesions are mainly pulmonary interstitial telangiectasia and have very little interstitial bleeding, and the lesions are low in proportion firstly and light secondly. Lesions of the lungs are likely to be artificially created at the time of sacrifice. The pathological changes of the stomach are small in proportion, and the chronic superficial inflammation is presented, in fact, the stomach inflammation of the mouse is relatively common. Increased gastric irritation due to inadequate chewing of food tends to cause inflammation. Therefore, the lesions are not considered to be caused by the medicament, so that the medicament has safety to mice.

In conclusion, the rhubarb compound composition provided by the invention has extremely strong inhibition effect on viruses such as HIV, SARS and the like, and has broad spectrum. Moreover, the safety of all the raw materials in the rhubarb compound composition is recorded in Chinese pharmacopoeia 2005 edition, the collocation is scientific and reasonable, the drug can adapt to the variability of viruses through the synergistic effect of multiple components on the premise of no side effect proved by toxicology, and pharmacodynamic experiments prove that the rhubarb compound composition has a remarkable inhibiting effect on the aspect of antivirus. Furthermore, the rhubarb compound composition can be prepared into various dosage forms, such as tablets, capsules, oral liquid and the like, on the premise of simply changing the dosage form process, and can be conveniently used under various conditions.

Claims

1. The application of the antiviral compound composition in resisting HIV virus, SARS virus and leukemia virus is characterized in that the composition consists of 7-110g of at least one component for detoxifying, resisting bacteria and diminishing inflammation, 5-60g of at least one component for tonifying qi and nourishing blood and 3-85g of at least one component for tonifying kidney and supporting yang.

2. The use of claim 1, wherein the detoxifying, antibacterial and anti-inflammatory component is at least one selected from the group consisting of extracts of honeysuckle, agrimony, licorice, trichosanthes root, isatis root, houttuynia cordata, forsythia suspensa and anthraquinone.

3. The use of claim 1, wherein the qi-invigorating and blood-nourishing component is at least one selected from fructus Jujubae, radix astragali, rhizoma Polygonati, rhizoma Atractylodis Macrocephalae, radix Codonopsis and radix Angelicae sinensis.

4. The use of claim 1, wherein the kidney-tonifying and yang-supporting component is at least one selected from the group consisting of epimedium, acanthopanax, yam, wolfberry, external-use plaster, eucommia, cistanche and bupleurum.

5. The use of any one of claims 1 to 4, wherein the composition comprises 4 to 20 parts of cistanche salsa, 3 to 30 parts of rhubarb anthraquinone extract, 5 to 35 parts of houttuynia cordata, 5 to 30 parts of isatis root, 2 to 20 parts of bupleurum, 5 to 30 parts of codonopsis pilosula, 3 to 25 parts of angelica sinensis and 1 to 15 parts of liquorice.

6. The use of any one of claims 1 to 4, wherein the composition comprises 5 to 35 parts of rhizoma Polygonati, 4 to 20 parts of cortex Eucommiae, 3 to 30 parts of anthraquinone extract of radix et rhizoma Rhei, 5 to 35 parts of flos Lonicerae, 1 to 15 parts of radix Glycyrrhizae, 5 to 30 parts of radix Trichosanthis, and 4 to 25 parts of barcan day.

7. The application of the antiviral compound composition in HIV virus, SARS virus and leukemia virus is characterized in that the composition comprises 7-110g of at least one component for detoxifying, resisting bacteria and diminishing inflammation, 5-60g of at least one component for tonifying qi and nourishing blood, 3-85g of at least one component for tonifying kidney and supporting yang and 7-75g of at least one component for ventilating lung and relieving cough.

8. The use of claim 7, wherein the lung-ventilating and cough-relieving component is at least one selected from the group consisting of platycodon grandiflorum, adenophora stricta, fritillaria cirrhosa, rhodiola rosea and ophiopogon japonicus.

9. The use of claim 7 or 8, wherein the composition comprises 3-30g of anthraquinone extract of rhubarb, 10-60g of agrimony, 3-25g of rhodiola rosea, 10-30g of astragalus membranaceus, 3-30g of herba epimedii, 5-30 g of radix ophiopogonis, 2-20 g of fritillary bulb and 5-30 g of fructus ziziphi chinensis.

10. The use of claim 7 or 8, wherein the composition comprises 3-30 parts of rhubarb anthraquinone extract, 4-30 parts of forsythia, 2-20 parts of platycodon grandiflorum, 5-30 parts of acanthopanax senticosus, 10-40 parts of Chinese yam, 5-25 parts of medlar, 5-30 parts of adenophora stricta and 5-25 parts of bighead atractylodes rhizome.

11. The use of claim 2, wherein the anthraquinone-based extract of rhubarb comprises at least one of chrysophanol, emodin, physcion, aloe-emodin and rhein.

12. The use of claim 1 or 7, wherein the antiviral compound composition is prepared by the following preparation method:

13. The use as claimed in claim 1 or 7, wherein the effective component of the antiviral compound composition is administered in an amount of 0.1-2 g/kg/d.